Haemophilus

Bull Soc Pathol Exot, 2004 May, 97(2), 100 - 3
{Haemophilus influenzae, the second cause of bacterial meningitis in children in Madagascar}; Razafindralambo M et al.; The Haemophilus influenzae b is one of the main germs causing bacterial meningitis in children in countries where the vaccine anti-Haemophilus influenzae b is not widely used . In Madagascar, no epidemiological study on this germ has been carried out . The objective of this research is to assess the role of Haemophilus influenzae meningitis in Antananarivo and to determine its epidemiological aspects and evolution . A multicentric study coordinated by the Institut Pasteur de Madagascar included all children less than 15 years old with infectious syndromes associated to a syndrome of meningial irritation and/or convulsion and/or coma . These children were admitted in the pediatric service of the three main hospitals in Antananarivo from June 1998 and June 2000 . A lumbar puncture was performed on each child; the cerebrospinal fluid was set aside for cytobacterial and biochemical controls completed with an antimicrobial sensitivity testing and a soluble antigens research . Out of 160 case studies, the Haemophilus influenzae b arrives at the second place among the agents causing bacterial meningitis in children . This type of bacteria is the source of 32% of meningitis after the Streptococcus pneumoniae (34%) . It affects 96% of children less than two years old, with a maximal frequency before the age of one year . The lethality rate is 28.6% and the neurological sequelae were observed in 31.4% of patients . Haemophilus influenzae is sensitive to the third generation cephalosporins but shows high resistance to chloramphenicol (42%), amoxicillin (29%) and gentamicin (22%) . The relatively high frequency as well as the high lethality rate caused by the Haemophilus influenzae b meningitis, affecting selectively the children under two years old, bring in the need to introduce the anti-Haemophilus influenzae b vaccine in the national vaccination program in Madagascar . This vaccine has proved to be efficient in many countries where it has been used . Furthermore, in the probabilistic treatment of bacterial meningitis in children, the third generation cephalosporins should be used in the first place.

Proc Natl Acad Sci U S A, 2004 Jul 20, 101(29), 10644 - 9 Epub 2004 Jul 09.
Protozoan predation, diversifying selection, and the evolution of antigenic diversity in Salmonella; Wildschutte H et al.; Extensive population-level genetic variability at the Salmonella rfb locus, which encodes enzymes responsible for synthesis of the O-antigen polysaccharide, is thought to have arisen through frequency-dependent selection (FDS) by means of exposure of this pathogen to host immune systems . The FDS hypothesis works well for pathogens such as Haemophilus influenzae and Neisseria meningitis, which alter the composition of their O-antigens during the course of bloodborne infections . In contrast, Salmonella remains resident in epithelial cells or macrophages during infection and does not have phase variability in its O-antigen . More importantly, Salmonella shows host-serovar specificity, whereby strains bearing certain O-antigens cause disease primarily in specific hosts; this behavior is inconsistent with FDS providing selection for the origin or maintenance of extensive polymorphism at the rfb locus . Alternatively, selective pressure may originate from the host intestinal environment itself, wherein diversifying selection mediated by protozoan predation allows for the continued existence of Salmonella able to avoid consumption by host-specific protozoa . This selective pressure would result in high population-level diversity at the Salmonella rfb locus without phase variation . We show here that intestinal protozoa recognize antigenically diverse Salmonella with different efficiencies and demonstrate that differences solely in the O-antigen are sufficient to allow for prey discrimination . Combined with observations of the differential distributions of both serotypes of bacterial species and their protozoan predators among environments, our data provides a framework for the evolution of high genetic diversity at the rfb locus and host-specific pathogenicity in Salmonella.

Diagn Microbiol Infect Dis, 2004 Jul, 49(3), 201 - 9
In vitro activity of tigecycline (GAR-936) tested against 11,859 recent clinical isolates associated with community-acquired respiratory tract and gram-positive cutaneous infections; Fritsche TR et al.; Tigecycline is a novel 9-t-butylglycylamido derivative of minocycline that has demonstrated activity against a variety of bacterial pathogens, including resistant isolates, during preclinical studies . In vitro activities of tigecycline and comparators were tested against 11,859 recent (2000 and 2002) bacterial strains recovered from patients in 29 countries with community-acquired respiratory tract disease (3,317 gram-positive and -negative strains) and skin and soft tissue infections (8,542 gram-positive strains) . All oxacillin-susceptible and -resistant Staphylococcus aureus (5,077 strains; tigecycline MIC(90), 0.5 microg/mL) and coagulase-negative staphylococci (1,432 strains; MIC(90), 0.5 microg/mL), penicillin-susceptible and -resistant Streptococcus pneumoniae (1,585 strains; MIC(90), < or =0.25 microg/mL), viridans group streptococci (212 strains; MIC(90), < or =0.25-0.5 microg/mL), vancomycin-susceptible and -resistant enterococci (1,416 strains; MIC(90), 0.25-0.5 microg/mL), beta-haemolytic streptococci (405 strains; MIC(90), < or =0.25 microg/mL), beta-lactamase positive and negative Haemophilus influenzae (1,220 strains; MIC(90), 1 microg/mL), Moraxella catarrhalis (495 strains; MIC(90), 0.25 microg/mL), and Neisseria meningitidis (17 strains; MIC(90), < or =0.12 microg/mL) were inhibited by 2 microg/mL or less of tigecycline . Whereas potency of tetracycline and doxycycline markedly dropped in various resistant organism subsets, tigecycline was unaffected with an overall MIC(90) of 0.5 microg/mL . These findings confirm that tigecycline maintains a truly broad spectrum like the tetracycline class while enhancing potency . It also incorporates stability to the commonly occurring tetracycline resistance mechanisms, making it an attractive candidate for continued clinical development against pathogens causing serious community-acquired respiratory tract infections, as well as cutaneous infections.

Wien Med Wochenschr, 2004 May, 154(9-10), 218 - 25
{Vaccination of the immunocompromised host}; Ullmann AJ et al.; Vaccinations are safe and effective in immunocompromised patients . Apparently most vaccines in this patient population are underutilized . General vaccination recommendations are expressed for influenza, diphtheria and tetanus . Pneumococcal, meningococcal und Haemophilus influenzae B immunizations are specially indicated for patients with or developing B-cell-deficiency . Live attenuated vaccines are usually contraindicated . The efficacy of the immunization and its indication can be additionally measured by antibody response, which is usually decreased compared to healthy subjects . Immunocompromised hosts with cancer will benefit from consistent immunization practices . Further clinical trials are urgently warranted.

J Clin Microbiol, 2004 Jul, 42(7), 3065 - 72
Prevalence of the hifBC, hmw1A, hmw2A, hmwC, and hia Genes in Haemophilus influenzae Isolates; Ecevit IZ et al.; Adherence of Haemophilus influenzae to respiratory epithelial cells is the first step in the pathogenesis of H . influenzae infection and is facilitated by the action of several adhesins located on the surface of the bacteria . In this study, prevalences of hifBC, which represent the pilus gene cluster; hmw1A, hmw2A, and hmwC, which represent high-molecular-weight (HMW) adhesin genes; and hia, which represents H . influenzae adhesin (Hia) genes were determined among clinical isolates of encapsulated type b (Hib) and nonencapsulated (NTHi) H . influenzae . hifBC genes were detected in 109 of 170 (64%) Hib strains and in 46 of 162 (28%) NTHi isolates (P = 0.0001) and were more prevalent among the invasive type b strains than invasive NTHi strains (P = 0.00003) . Furthermore, hifBC genes were significantly more prevalent (P = 0.0398) among NTHi throat isolates than NTHi middle ear isolates . hmw1A, hmw2A, hmwC, and hia genes were not detected in Hib strains . Among NTHi isolates, the prevalence of hmw1A was 51%, the prevalence of hmw2A was 23%, the prevalence of hmwC was 48%, and the prevalence of hia was 33% . The hmw genes were significantly more prevalent among middle ear than throat isolates, while hia did not segregate with a respiratory tract site . These results show the variability of the presence of adhesin genes among clinical H . influenzae isolates and suggest that hemagglutinating pili may play a larger role in H . influenzae nasopharyngeal colonization than in acute otitis media whereas the HMW adhesins may be virulence factors for acute otitis media.

J Antimicrob Chemother, 2004 Aug, 54(2), 393 - 400 Epub 2004 Jul 08.
Inability of L22 ribosomal protein alteration to increase macrolide MICs in the absence of efflux mechanism in Haemophilus influenzae HMC-S; Peric M et al.; BACKGROUND: Haemophilus influenzae HMC-C with high-level macrolide resistance after multi-step selection by clarithromycin reverted spontaneously and became hypersusceptible to macrolides . OBJECTIVE: Determination of macrolide resistance mechanism(s) in hypersusceptible and hyperresistant strains . METHODS: The presence of macrolide efflux in the strains was studied by radioactive erythromycin accumulation . Ribosomal mutations were investigated by sequencing . The possible role of acrAB clusters in macrolide resistance was studied by sequencing and expression analysis . RESULTS: The parent strain had no ribosomal alteration, but both high-level resistant and hypersusceptible strains had R88P mutations in ribosomal protein L22 . Radioactive macrolide accumulation studies pointed to the presence of macrolide efflux in the high-level resistant and parent strains, but not in the hypersusceptible derivative . Transformation of hypersusceptible strains using total DNA from the parent strain restored the macrolide efflux system in the hypersusceptible strain, which was confirmed by MIC levels and radioactive erythromycin accumulation similar to that of the mutant resistant strain . Analysis of sequence and transcription of acrAB gene clusters showed no significant differences between resistant and hypersusceptible derivatives . CONCLUSION: Mutation in ribosomal protein L22 alone does not confer high-level macrolide resistance unless efflux is present.

Appl Environ Microbiol, 2004 Jul, 70(7), 4136 - 43
Use of the riboflavin synthase gene (ribC) as a model for development of an essential gene disruption and complementation system for Haemophilus influenzae; Saeed-Kothe A et al.; We have developed a system for rapid and reliable assessment of gene essentiality in Haemophilus influenzae Rd strain KW20 . We constructed two "suicide" complementation vectors (pASK5 and pASK6) containing 5' and 3' regions of the nonessential ompP1 gene flanking a multiple cloning site and a selectable marker (a chloramphenicol resistance gene or a tetracycline resistance cassette) . Transformation of H . influenzae with the complementation constructs directs chromosomal integration of a gene of interest into the ompP1 locus, where the strong, constitutive ompP1 promoter drives its expression . This single-copy, chromosome-based complementation system is useful for confirming the essentiality of disrupted genes of interest . It allows genetic analysis in a background free of interference from any upstream or downstream genetic elements and enables conclusive assignment of essentiality . We validated this system by using the riboflavin synthase gene (ribC), a component of the riboflavin biosynthetic pathway . Our results confirmed the essentiality of ribC for survival of H . influenzae Rd strain KW20 and demonstrated that a complementing copy of ribC placed under control of the ompP1 promoter reverses the lethal phenotype of a strain with ribC deleted.

J Pediatr, 2004 Jul, 145(1), 58 - 66
Safety of DTaP-based combined immunization in very-low-birth-weight premature infants: frequent but mostly benign cardiorespiratory events; Pfister RE et al.; OBJECTIVE: To evaluate the safety of diphtheria-tetanus-acellular pertussis-inactivated polio-Haemophilus influenzae type B (DTaP-IPV-HIB) immunization in premature infants . STUDY DESIGN: Observational study of 78 very low birth weight premature infants (mean gestational age, 28+/-2 weeks; mean birth weight, 1045+/-357 g) given DTaP-IPV-HIB vaccine before hospital discharge . Apnea, bradycardia, oxygen requirements and saturation, feeding practice, and medical interventions were assessed before and after immunization . The results were analyzed by the severity of the clinical condition and the persistence of prematurity-associated symptoms . RESULTS: Administration of DTaP-IPV-HIB elicited resurgence or increase in cardiorespiratory events in 47% of infants (15% had apnea, 21% had bradycardia, 42% of desaturations) . Most vaccine-triggered events resolved spontaneously or after brief stimulation . The relative risk was 5- to 8-fold higher in infants with a severe clinical course or persistence of cardiorespiratory symptoms at the time of immunization . Bag-mask respiratory support was given to 5 of 78 infants, and O(2) requirements increased transiently in 4 of 21 infants with chronic lung disease, none requiring reventilation . Reintroduction of O(2) supplementation, interruption of active oral feeding, or postponing of hospital discharge was not required . CONCLUSIONS: Cardiorespiratory events were frequently increased after DTaP-IPV-HIB immunization, requiring monitoring and appropriate intervention . However, these episodes did not have detrimental impact on the infants' clinical course . Timely immunization is warranted even in the most vulnerable preterm infants.

Electrophoresis, 2004 Jul, 25(13), 1949 - 63
Capillary electrophoresis-mass spectrometry for glycoscreening in biomedical research; Zamfir A et al.; Application of capillary electrophoresis (CE) in combination with mass spectrometry (MS) and tandem MS to glycoscreening in biomedical projects is highlighted . In the first part recent CE-MS experiments by sheath liquid CE and multiple stage MS are reported . Neutral and negatively charged N-glycan mixtures from ribonuclease B and fetuin, high-mannose type N-glycoforms, oligosaccharides from lipopolysaccharides (LPS) of Haemophilus influenzae, polysaccharides of Pseudomonas aeruginosa and Staphylococcus aureus were analyzed . A particular emphasis is devoted to the applicability of novel off- and on-line CE-MS and tandem MS methods for screening of proteoglycan-derived oligosaccharides, glycosaminoglycans (GAGs), such as hyaluronates from Streptococcus agalactiae, chondroitin/dermatan sulfates (CS/DS) from bovine aorta and human skin fibroblast decorin, and heparin/heparan sulfate (HS) from porcine and bovine mucosa . The performance of CE-MS/MS for identification of glycoforms in glycopeptides and glycoproteins is illustrated by experiments performed on complex mixtures from urine of patients suffering from a hereditary N-acetylhexosaminidase deficiency (Schindler's disease) and urine of patients suffering from cancer cachexia . For determination of glycosylation patterns in glycoproteins like enzymes and antibodies by CE/MS, both CE-matrix assisted laser desorption/ionization (MALDI) and CE-electrospray ionization (ESI)-MS were functional . Finally, the potential of CE-ESI-MS strategy in glycolipid analysis is demonstrated for gangliosides from bovine brain for which particular CE buffer conditions are required.

J Infect, 2004 Aug, 49(2), 115 - 25
Activity of telithromycin against key pathogens associated with community-acquired respiratory tract infections; Low DE et al.; OBJECTIVES: To investigate the correlation between in vitro susceptibility of isolates and clinical outcomes with telithromycin in respiratory tract infections . METHODS: The activity of telithromycin was determined by in vitro susceptibility testing of key respiratory tract pathogens isolated from patients with community-acquired pneumonia, acute exacerbations of chronic bronchitis or acute maxillary sinusitis enrolled in 14 Phase III/IV clinical trials evaluating the clinical efficacy of telithromycin . RESULTS: In this pooled analysis, telithromycin mode minimum inhibitory concentration (MIC) and MIC90, respectively, were: 0.016 and 0.03 mg/l against Streptococcus pneumoniae (n=626); 0.03 and 0.5 mg/l for penicillin-resistant S . pneumoniae (n=56); 0.03 and 1 mg/l for erythromycin-resistant S . pneumoniae (n=81); 2 and 4 mg/l against Haemophilus influenzae (including beta-lactamase producers; n=627); both 0.12 mg/l for Moraxella catarrhalis (n=159) and both 0.25 mg/l for Staphylococcus aureus (n=124) . Telithromycin (5 or 7-10 days) resulted in overall clinical and bacteriologic success rates of 88.1% (1593/1808) and 89% (1593/1789), respectively . CONCLUSIONS: High levels of in vitro susceptibility to telithromycin are paralleled by high rates of clinical cure and bacteriologic eradication.

J Infect, 2004 Aug, 49(2), 109 - 14
Surveillance of the susceptibility of ocular bacterial pathogens to the fluoroquinolone gatifloxacin and other antimicrobials in Europe during 2001/2002; Morrissey I et al.; OBJECTIVES: To determine the identity of European ocular bacterial pathogens and their susceptibility to topical antimicrobial agents . METHODS: Bacterial isolates derived from clinically significant ocular infections were collected from 10 European centres . Bacteria were re-identified and susceptibility to gatifloxacin, ciprofloxacin, ofloxacin, fusidic acid, gentamicin and chloramphenicol was determined using the NCCLS agar incorporation method at a central testing laboratory . RESULTS: Five hundred and thirty-two isolates were submitted for analysis . The most common pathogen was methicillin-susceptible Staphylococcus aureus, followed by Haemophilus influenzae and Streptococcus pneumoniae . Gatifloxacin was the most potent antimicrobial agent tested for isolates from each European country as measured by pure MIC or percentage resistance (using 95% confidence intervals) . Only methicillin-resistant S . aureus was in any way refractory to the action of gatifloxacin and other fluoroquinolones . CONCLUSIONS: Fluoroquinolones offer broad-spectrum coverage for the treatment of ocular pathogens . Gatifloxacin ophthalmic solution should be a significant improvement on currently available fluoroquinolones mainly due to enhanced activity against streptococci and coagulase-negative staphylococci (which accounted for almost one-third of ocular pathogens).

Arq Neuropsiquiatr, 2004 Jun, 62(2A), 250 - 2 Epub 2004 Jun 23.
Etiology of bacterial meningitis among children aged 2-59 months in Salvador, Northeast Brazil, before and after routine use of Haemophilus influenzae type B vaccine; Nascimento-Carvalho CM et al.; OBJECTIVE: To describe the frequency of etiologic agents of bacterial meningitis (BM) among children aged 2-59 months in a sample of patients in Salvador, Northeast Brazil, with emphasis on the frequency of BM of unknown etiology (BMUE), just before, during and after the implementation of routine immunization of infants with Haemophilus influenzae type b (Hib) vaccination . METHOD: Demographic, clinical and cerebrospinal fluid (CSF) information was collected from the chart of every patient, aged 2-59 months, whose CSF exam was performed at the CSF Lab - Jose Silveira Foundation, between January 1989 and December 2001 . Every CSF exam was completely performed according to standard methods . The etiologic diagnosis was based on either culture and/or latex-agglutination test . When the agent was only seen on Gram stained smear, the diagnosis was descriptive . BMUE was defined as: glucose < 40mg / dl, protein > 100 mg / dl, white blood cell count > 20 cells / mm(3), percentage of neutrophils > 80% . RESULTS: Of 1519 patients, 894 (58.9%) had normal exams and BM was diagnosed in 95 (6.2%) . Etiologic agents were: Hib (44.2%), meningococcus (13.7%), Gram-negative bacilli (11.6%), Mycobacterium tuberculosis (6.3%), pneumococcus (4.2%), other agents (4.2%); BMUE was diagnosed in 15.8% of cases with BM . By analysing the frequency of BMUE and Hib among all exams performed yearly, the peaks were recorded in 1989 (5.3%) and 1990 (16.9%), respectively, decreasing to 0.7% and 0% in 2001 . CONCLUSION: It is possible that the implementation of the conjugate Hib vaccine during the 1990's has been decreasing not only the occurrence of Hib meningitis but also of BMUE.

APMIS, 2004 Apr-May, 112(4-5), 299 - 303
Antibody response to the patient's own Haemophilus influenzae isolate can support the aetiology in lower respiratory tract infections; Stralin K et al.; In order to understand the clinical importance of Haemophilus influenzae isolated from sputum samples, an indirect immunofluorescence (IF) assay was developed, using the patient's own isolate as the antigen . The method was tested on samples from six patients with lower respiratory tract infection (LRTI) and H . influenzae isolated from blood (n=2), sputum (n=3) or both (n=1), and on two healthy adults with H . influenzae isolated from the nasopharynx . Between acute and convalescent sera, a four-fold IgG antibody increase was achieved in five of six LRTI patients, including the three blood culture-positive patients . One LRTI patient and the two asymptomatic carriers showed stable antibody levels against their own isolate . Although small, the study indicates that indirect IF can be a promising tool for determining whether a H . influenzae strain represents the probable cause of infection or just a strain colonising the airways . More extensive studies should be performed in order to establish the usefulness of the assay.

J Trop Pediatr, 2004 Jun, 50(3), 158 - 63
The causes of hospital admission and death among children in Bamako, Mali; Campbell JD et al.; The health burden and mortality caused by infections during childhood remain large in sub-Saharan Africa . We performed a review of the causes of hospitalization and death among children admitted to a pediatric teaching hospital in Bamako, Mali . Medical records of children admitted throughout 2000 were systematically sampled and abstracted for demographics, diagnosis, hospital course, and disposition . A sample of 1644 charts, from 5001 admitted children, were abstracted . The median age was 8 months . Half of the children had a febrile illness . All diagnoses were made clinically . The annual incidence per 100,000 and case fatality rates of the four most common serious infections, excluding malaria, were as follows: pneumonia, 165 (12 per cent); sepsis, 75 (37 per cent); meningitis, 71 (20 per cent); and enteric fever, 14 (12 per cent) . An estimated 1300 children were admitted with thick-smear confirmed malaria; at least 64 per cent met World Health Organization criteria for severe malaria and 11 per cent died . Seventy-one per cent of admissions were due to infections . Overall 21 per cent of children admitted died in the hospital, most within the first 3 days of admission . Infectious diseases remain the primary cause of hospitalization among Malian children and frequently lead to death . A substantial proportion of this morbidity and mortality is probably attributable to vaccine-preventable diseases, such as Haemophilus influenzae type B, Streptococcus pneumoniae, and Neisseria meningitidis . Prospective surveillance using microbiological data is needed to delineate the organism-specific burdens.

J Trop Pediatr, 2004 Jun, 50(3), 131 - 6
Haemophilus type B meningitis in Saudi children under 5 years old; Al-Mazrou YY et al.; This study was designed to determine the magnitude of bacterial meningitis in general and Hib meningitis in particular among children below the age of 5 years . A population-based, prospective descriptive and analytical study was conducted in five regions, one each in northern, southern, eastern, western, and central parts of Saudi Arabia . Active surveillance for cases of bacterial meningitis among the study population, which comprised 171,818 children under 5 years of age, was implemented . A total of 208 cases of meningitis were identified, of which 141 (67.8 per cent) were identified with a definite causative organism . The remaining 67 cases (32 per cent) were labeled as aseptic meningitis . The overall incidence of meningitis was 60.53/10(5) in under-fives with a disease spectrum similar to that reported in studies conducted in other countries . The three leading causes of meningitis were Hib (Haemophilus influenzae type B), MCM (Neisseria menigitides) and SPN (Streptococcus pneumoniae) . Hib meningitis constituted 28 per cent of cases with an incidence rate of 16.88/10(5) children . There was a marked regional variation in Hib incidence . MCM was the second leading cause (18 per cent) of meningitis with an incidence of 10.77/10(5) while SPN comprised 11 per cent of cases and its incidence was 9.69/10(5) . Almost all MCM cases were related to meningitis outbreaks that occurred in Saudi Arabia during two successive Hajj seasons (2000-2001) . Hib cases showed a bimodal seasonality, one peak during March-May, the other during September-November . The fact that this study is the first national base-line data on meningitis in general and Hib incidence in particular, has augmented further justification for introducing Hib vaccine within the national Expanded Program on Immunization (EPI) . Based on the experience gained during this study regarding surveillance of meningitis disease, optimal methods to strengthen meningitis surveillance were identified . A model of Meningitis Diseases Surveillance was generated that can be tested and then generalized . The study has documented beyond doubt the impact of Hajj seasons on MCM disease occurrence and further justifies the rigorous control and preventive measures being taken in this aspect.

Pediatrics, 2004 Jul, 114(1), 187 - 95
Children who have received no vaccines: who are they and where do they live?
Smith PJ, Chu SY, Barker LE.
CONTEXT: Each year 2.1 million children 19 to 35 months of age are undervaccinated . Among these are children who have received no vaccinations . Unvaccinated children are at increased risk of acquiring and transmitting vaccine-preventable diseases . OBJECTIVES: To assess whether the characteristics of children with no vaccinations differ from those of undervaccinated children, to monitor trends in the numbers of unvaccinated children, and to identify states with high rates and counties with large numbers of unvaccinated children . DESIGN: A nationally representative probability sample of children 19 to 35 months of age was collected annually between 1995 and 2001 . Vaccination histories were ascertained from children's medical providers . Undervaccinated children had received > or =1 dose of diphtheria-tetanus-pertussis, polio, measles, Haemophilus influenzae type b, hepatitis B, or varicella vaccine but were not fully vaccinated . Unvaccinated children were children who were reported as having no medical providers and having received no vaccinations or children whose medical providers reported administering no vaccinations . PARTICIPANTS: A total of 151,720 children sampled between 1995 and 2001, 795 of whom were unvaccinated . RESULTS: Undervaccinated children tended to be black, to have a younger mother who was not married and did not have a college degree, to live in a household near the poverty level, and to live in a central city . Unvaccinated children tended to be white, to have a mother who was married and had a college degree, to live in a household with an annual income exceeding 75,000 dollars, and to have parents who expressed concerns regarding the safety of vaccines and indicated that medical doctors have little influence over vaccination decisions for their children . Unvaccinated children were more likely to be male than female . Annually, approximately 17,000 children were unvaccinated . The largest numbers of unvaccinated children lived in counties in California, Illinois, New York, Washington, Pennsylvania, Texas, Oklahoma, Colorado, Utah, and Michigan . States that allowed philosophical exemptions to laws mandating vaccinations for children as they entered school had significantly higher estimated rates of unvaccinated children . CONCLUSIONS: Unvaccinated children have characteristics that are distinctly different from those of undervaccinated children . Unvaccinated children are clustered geographically, increasing the risk of transmitting vaccine-preventable diseases to both unvaccinated and undervaccinated children.

J Bacteriol, 2004 Jul, 186(14), 4575 - 84
Mu-like prophage strong gyrase site sequences: analysis of properties required for promoting efficient mu DNA replication; Oram M et al.; The bacteriophage Mu genome contains a centrally located strong gyrase site (SGS) that is required for efficient prophage replication . To aid in studying the unusual properties of the SGS, we sought other gyrase sites that might be able to substitute for the SGS in Mu replication . Five candidate sites were obtained by PCR from Mu-like prophage sequences present in Escherichia coli O157:H7 Sakai, Haemophilus influenzae Rd, Salmonella enterica serovar Typhi CT18, and two strains of Neisseria meningitidis . Each of the sites was used to replace the natural Mu SGS to form recombinant prophages, and the effects on Mu replication and host lysis were determined . The site from the E . coli prophage supported markedly enhanced replication and host lysis over that observed with a Mu derivative lacking the SGS, those from the N . meningitidis prophages allowed a small enhancement, and the sites from the Haemophilus and Salmonella prophages gave none . Each of the candidate sites was cleaved specifically by E . coli DNA gyrase both in vitro and in vivo . Supercoiling assays performed in vitro, with the five sites or the Mu SGS individually cloned into a pUC19 reporter plasmid, showed that the Mu SGS and the E . coli or N . meningitidis sequences allowed an enhancement of processive, gyrase-dependent supercoiling, whereas the H . influenzae or Salmonella serovar Typhi sequences did not . While consistent with a requirement for enhanced processivity of supercoiling for a site to function in Mu replication, these data suggest that other factors are also important . The relevance of these observations to an understanding of the function of the SGS is discussed .

Proteins, 2004 Aug 15, 56(3), 564 - 71
Novel structure and nucleotide binding properties of HI1480 from Haemophilus influenzae: a protein with no known sequence homologues; Lim K et al.; The crystal structure of the Haemophilus influenzae protein HI1480 was determined at 2.1-A resolution . The amino acid sequence of HI1480 is unique, having no homology with other known protein sequences . The protein adopts a novel alpha+beta fold, and associates into a dimer of tightly associated dimers . The tight dimers are formed by intermolecular interactions that are mediated by an antiparallel beta-barrel involving both monomers . Helical regions of two dimers mediate the tetramer formation . The helical region contains a four-helix bundle that has been seen only in the anticodon binding domains of class I tRNA synthetases . A cluster of four residues, Tyr18, Arg134, Glu26, and Lys12 is located in a depression formed at the four-helix bundle/ beta-barrel interface . The arrangement is suggestive of an active center, possibly a catalytic site . The HI1480 gene is located within the Mu-like prophage region of H . influenzae, has no homology to bacteriophage genes, and is flanked by transposases . Hence, this is an example of horizontal transfer from an unknown organism . Gel mobility shift assays revealed that HI1480 binds DNA and RNA molecules . Double-stranded DNA is favored over single-stranded DNA, and longer DNA molecules are bound better than shorter ones .

Enferm Infecc Microbiol Clin, 2004 Jun-Jul, 22(6), 323 - 7
{Activity of telithromycin and other oral antibiotics against respiratory tract pathogens with acquired mechanisms of resistance}; Gomez-Garces JL et al.; INTRODUCTION: Antimicrobial resistance among strains of Streptococcus pneumoniae, Haemophilus influenza, and Streptococcus pyogenes has limited the usefulness of conventional agents for the treatment of some infectious diseases related with these microorganisms . There is thus a clear need for new antimicrobial agents active against these resistant strains . OBJECTIVES: To assess the in vitro susceptibility to telithromycin and other oral antimicrobial agents of 307 strains of S . pneumoniae, H . influenzae, and S . pyogenes with acquired mechanisms of resistance to macrolides and/or beta-lactams . METHODS: Antimicrobial susceptibility testing was performed by the agar dilution method, and results were preferentially evaluated according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines . RESULTS: The activity of telithromycin against erythromycin-resistant S . pneumoniae was excellent . All the strains were inhibited with 2 mg/l, regardless of the mechanism of macrolide resistance exhibited . Telithromycin was slightly more active than the tested macrolides in beta-lactamase-producing H . influenzae strains, with a MIC90 of 2 mg/l . Telithromycin also demonstrated potent activity against macrolide-resistant S . pyogenes . The MIC of telithromycin was > 4 mg/l for only one out of the 202 tested strains . CONCLUSIONS: Telithromycin is a very promising therapeutic option against bacterial pathogens of the respiratory tract, including strains that have developed mechanisms of resistance to macrolides and/or beta-lactams.

Mol Microbiol, 2004 Jul, 53(2), 651 - 64
Inactivation of deoxyadenosine methyltransferase (dam) attenuates Haemophilus influenzae virulence; Watson ME Jr et al.; Mutants in deoxyadenosine methyltransferase (dam) from many Gram-negative pathogens suggest multiple roles for Dam methylase: directing post-replicative DNA mismatch repair to the correct strand, guiding the temporal control of DNA replication and regulating the expression of multiple genes (including virulence factors) by differential promoter methylation . Dam methylase (HI0209) in strain Rd KW20 was inactivated in Haemophilus influenzae strains Rd KW20, Strain 12 and INT-1; restriction with Dam methylation-sensitive enzymes DpnI and DpnII confirmed the absence of Dam methylation, which was restored by complementation with a single copy of dam ectopically expressed in cis . Despite the lack of increased mutation frequency, the dam mutants had a 2-aminopurine-susceptible phenotype that could be suppressed by secondary mutations in mutS, suggesting a role for Dam in H . influenzae DNA mismatch repair . Invasion of human brain microvascular endothelial cells (HBMECs) and human respiratory epithelial cells (NCI-H292) by the dam mutants was significantly attenuated in all strains, suggesting the absence of a Dam-regulated event necessary for uptake or invasion of host cells . Intracellular replication was inhibited only in the Strain 12 dam mutant, whereas in the infant rat model of infection, the INT-1 dam mutant was less virulent . Dam activity appears to be necessary for both in vitro and in vivo virulence in a strain-dependent fashion and may function as a regulator of gene expression including virulence factors.

Klin Mikrobiol Infekc Lek, 2004 Jun, 10(3), 130 - 133
{Diagnostics of invasive meningococcal, haemophilus and pneumococcal disease by PCR assay}; Kalmusova J et al.; Objectives: Development of extended polymerase chain reaction (PCR) for non-culture detection of Nesseria meningitidis, Haemophilus influenzae and Streptococcus pneumonie from invasive infections . Materials and methods: A method of PCR was optimalised on strains of Nesseria meningitidis, Haemophilus influenzae b and Streptococcus pneumonie . Detection of pathogens was evaluated on 230 samples from patiens with invasive infection . Results: Positive results of PCR were found in 103 samples of 230 (44.7 %) . The percentage of positivity was higher in CSF samples (57.0 %) than in serum (33.8 %) or blood (33.3 %) samples . Conclusion: PCR method enables etiological diagnostics in cases, where antibiotic treatment was started . PCR results are available earlier than the results of cultivation . Multilocus sequence typing (MLST) of PCR products enables clonal analysis of etiological agents even in cases with negative results of cultivation.

Klin Mikrobiol Infekc Lek, 2004 Jun, 10(3), 118 - 123
{The effect of routine vaccination in the Czech Republic on the incidence of invasive diseases caused by Haemophilus influenzae b}; Krizova P et al.; Objectives: The paper is an analysis of the results of a five-year surveillance programme in the Czech Republic (1999-2003); it evaluates the efficacy of routine Hib vaccination that started in July 2001 . Material and methods: Morbidity due to Hib was explored in a surveillance programme, which since 2002 also included the investigation of Hib vaccine failure . Hib identification was carried out with standard methods, serotypes were verified using PCR, biotyping was carried out in all strains and in selected strains also multilocus sequential typing . Results: In the years 1999-2003 invasive Hib disease presented mostly as meningitis, followed by epiglottitis . Mortality due to an invasive Hib disease was in the years 1999-2003 2.3 % . Among the Hib strains isolated in invasive disorders predominated the biotype I and the sequence type ST-6 . Conclusions: Following the introduction of routine Hib vaccination in the Czech Republic there was an overall drop in morbidity due to Hib invasive disorders . This was most obvious in a decrease in the morbidity of Hib meningitis and in the vaccinated age group . Two years after the introduction of routine Hib vaccination morbidity dropped by 81 % in children aged 0 to 1 year . In higher age groups there was no change in the number of invasive Hib disease . Neither was there an increase in "non-b" haemophilus invasive disorders . Failure of Hib vaccination is a rare occurence.

J Biol Chem, 2004 Aug 27, 279(35), 36171 - 4 Epub 2004 Jun 28.
NF-kappaB is essential for induction of CYLD, the negative regulator of NF-kappaB: evidence for a novel inducible autoregulatory feedback pathway; Jono H et al.; The transcription factor NF-kappaB regulates genes involved in inflammatory and immune responses, tumorigenesis, and apoptosis . In contrast to the pleiotropic stimuli that lead to its positive regulation, the known signaling mechanisms that underlie the negative regulation of NF-kappaB are very few . Recent studies have identified the tumor suppressor CYLD, loss of which causes a benign human syndrome called cylindromatosis, as a key negative regulator for NF-kappaB signaling by deubiquitinating tumor necrosis factor (TNF) receptor-associated factor (TRAF) 2, TRAF6, and NEMO (NF-kappaB essential modulator, also known as IkappaB kinase gamma) . However, how CYLD is regulated remains unknown . The present study revealed a novel autoregulatory feedback pathway through which activation of NF-kappaB by TNF-alpha and bacterium nontypeable Haemophilus influenzae (NTHi) induces CYLD that in turn leads to the negative regulation of NF-kappaB signaling . In addition, TRAF2 and TRAF6 appear to be differentially involved in NF-kappaB-dependent induction of CYLD by TNF-alpha and NTHi . These findings provide novel insights into the autoregulation of NF-kappaB activation.

Int J Antimicrob Agents, 2004 Jul, 24(1), 18 - 23
Use of oral cephalosporins in the treatment of acute otitis media in children; Brook I; The selection of the most effective antimicrobial to treat acute otitis media (AOM) has become more difficult in recent years because of increasing antibiotic resistance among all AOM pathogens . Resistance of Streptococcus pneumoniae to penicillin as well as amoxicillin ranges from 30 to 55% in the USA . Currently, 40-55% of Haemophilus influenzae and 90-100% of Moraxella catarrhalis are resistant to penicillin because of the production of Beta-lactamases . This review discusses the availability of oral cephalosporins that can be utilised for the treatment of AOM in children . An evaluation is made regarding their in vitro activity against the pathogens, their middle-ear concentrations, pharmacokinetics and pharmacodynamics (PK/PD) . The cephalosporins that will be discussed are cefuroxime-axetil, cefprozil, cefdinir and cefpodoxime-proxetil . The current recommendations for therapy of AOM limit the choices of clinicians to a single cephalosporin (cefuroxime-axetil) . However, clinical, bacteriological and PK/PD data shows that several other cephalosporins (cefprozil, cefdinir and cefpodoxime-proxetil) possess similar indices which provide the clinician with wider therapeutic choices that can insure better compliance and ultimately better success in eradication of the infection.

Med Trop (Mars), 2004, 64(1), 33 - 8
{Role of thiamphenicol in the treatment of community-acquired lung infections}; Raymond J et al.; Streptococcus pneumoniae and Haemophilus influenzae are the two main pathogens responsible for bacterial respiratory tract infections . Their antimicrobial susceptibility to antibiotics like beta-lactams, macrolides or fluoroquinolones has been largely studied, while it remains less known to other antibiotics like thiamphenicol, erythromycin, cotrimoxazole or tetracycline, often used in developing countries due to their availability . In this study, the activity of chloramphenicol and thiamphenicol on different respiratory tract pathogens was found to be equivalent . However, thiamphenicol was better in detecting resistant organisms . One hundred S . pneumoniae among which 69% had reduced susceptibility to penicillin (PRSP) and 87 H . influenzae isolates, 39.1% producing beta-lactamase, were recovered from sputum cultures in children . All H . influenzae and all penicillin susceptible S . pneumoniae strains were sensitive to thiamphenicol . Susceptibility of penicillin sensitive S . pneumoniae to erythromycin, cotrimoxazole and tetracycline was 70.9%, 83.9%, and 90.3% respectively . Susceptibility of PRSP to thiamphenicol, erythromycin, cotrimoxazole and tetracycline was 68.1%, 7.2%, 17.4% and 44.9% respectively . Thiamphenicol and chloramphenicol are still active against respiratory pathogens.

Biometals, 2004 Jun, 17(3), 235 - 43
Lactoferrin receptors in gram-negative bacteria: insights into the iron acquisition process; Ekins A et al.; One component of the anti-microbial function of lactoferrin (Lf) is its ability to sequester iron from potential pathogens . To overcome this iron limitation, a number of gram-negative bacterial pathogens have developed a mechanism for acquiring iron directly from this host glycoprotein . This mechanism involves surface receptors capable of specifically binding Lf from the host, removing iron and transporting it across the outer membrane . The iron is then bound by a periplasmic iron-binding protein, FbpA, and transported into the cell via an inner membrane complex comprised of FbpB and FbpC . The receptor has been shown to consist of two proteins, LbpA and LbpB . LbpB is bilobed lipoprotein anchored to the outer membrane via fatty acyl groups attached to the N-terminal cysteine . LbpA is a homologue of siderophore receptors, which consist of an N-terminal plug and a C-terminal beta-barrel region . We propose that the receptor proteins, LbpA and LbpB, induce conformational changes in human Lf (hLf) that lower its affinity for iron that binding by FbpA can drive the transport across the outer membrane, a mechanism shared with transferrin (Tf) receptors . The interaction between the receptor proteins and Lf is quite extensive and has been previously studied by using chimeric proteins comprised of Lf & Tf . In an attempt to evaluate the role of FbpA in the transport process, a series of site-directed mutants of FbpA were prepared and used to replace the wild-type protein in the iron acquisition pathway . The mutations were made in the iron-binding and anion-binding ligands of FbpA and were designed to result in altered binding properties . Protein crystallography of the iron-bound form of the Q58L mutant protein revealed that it was in the open conformation with iron coordinated by Y195 and Y196 from the C-terminal domain but not by the other iron-liganding amino acids from the N-terminal domain, H9 and E57 . Replacement of the native FbpA in Neisseria meningitidis with wild-type or mutant Haemophilus influenzae FbpAs resulted in a defect in growth on Tf or Lf, suggesting that there may be a barrier to functional expression of H . influenzae FbpAs in Neisseria meningitidis . Thus mutants of the N . meningitidis FbpA are being prepared to replace wild-type protein in order to test their ability to mediate transport from hLf.

J Microbiol Immunol Infect, 2004 Jun, 37(3), 164 - 8
Epidemiology of Haemophilus influenzae type b meningitis in Taiwan, 1997 and 2000; Shao PL et al.; In order to determine the incidence of Haemophilus influenzae type b (Hib) meningitis in Taiwan, we analyzed the National Health Insurance Research Database (NHIRD) . We identified cases that were younger than 5 years old and diagnosed with Hib meningitis from the NHIRD in both 1997 and 2000 . Sixteen and 8 children with Hib meningitis were identified in the NHIRD for 1997 and 2000, respectively . The majority of cases were 2 years old and under . The overall fatality rate was 8.33% . Assuming similar proportions of Hib meningitis in cases of meningitis with identified causes and meningitis without identified causes, the annual incidence of Hib meningitis in children younger than 5 years old was estimated to be 5.57 and 3.22 per 100,000 in years 1997 and 2000, respectively . The incidence of Hib meningitis in Taiwan is lower than in western countries and other Asian countries . This is consistent with the notion that Hib meningitis incidence is low in Chinese populations . The decline in incidence from 1997 to 2000 is likely a result of the introduction of a self-paid Hib vaccine in Taiwan, starting in 1993 . The availability of the NHIRD provides an extra tool for the epidemiological study of infectious diseases in Taiwan and can be valuable for similar studies.

Pulm Pharmacol Ther, 2004, 17(4), 199 - 203
Ertapenem as initial antimicrobial monotherapy for patients with chronic obstructive pulmonary disease hospitalized with typical community-acquired pneumonia; Friedland IR et al.; This report describes a post-hoc analysis of two large studies of typical community-acquired pneumonia (CAP) in hospitalized patients, focusing on demographics, disease characteristics, and outcome in patients with and without chronic obstructive pulmonary disease (COPD) . In both studies, ertapenem 1 g IV daily was compared with ceftriaxone 1 g IV daily as initial antimicrobial therapy . Clinically improving patients could be switched to oral antibiotic therapy after 3 days . Of the 857 patients treated in both studies, 264 (31%) had COPD . The proportions of patients who were male, were >/=65 years of age, had a Pneumonia Severity Index of IV/V, or had Haemophilus influenzae isolated in a baseline culture were higher in patients with COPD . Streptococcus pneumoniae was the most common pathogen both in patients with and without COPD . Clinical response rates in assessable patients 7-14 days after completion of therapy for the combined treatment groups were 90% (187/208) for patients with COPD and 93% (424/456) for those without COPD (odds ratio 0.7 {95% CI, 0.4-1.2}, P = 0.17) . Of assessable COPD patients, 109/121 (90%) treated with ertapenem and 78/87 (90%) treated with ceftriaxone achieved a favorable clinical response (odds ratio 1.0 {95% CI, 0.6-1.8}, P = 0.94) . The outcome in patients with or without COPD was similar regardless of therapy . In patients with COPD as well as in the overall study population, the efficacy of ertapenem as initial antimicrobial monotherapy for patients with serious typical community-acquired pneumonia was comparable to that of ceftriaxone.

Jpn J Antibiot, 2004 Apr, 57(2), 187 - 95
{The isolation frequency and antimicrobial susceptibility of Haemophilus influenzae isolated in Saga University Hospital}; Fukutomi Y et al.; Isolation frequency and antimicrobial susceptibility of Haemophilus influenzae isolated in Saga University hospital from October 2002 to September 2003 were investigated . Out of 155 H . influenzae strains subjected 77 were isolated from pediatrics specimens . beta-Lactamase negative ampicillin (ABPC)-resistant H . influenzae (BLNAR), against which MICs of ABPC were higher than 4 microg/mL, were 32 strains (20.6%), and it became 63 strains (41.3%) when Low-BLNAR, against which MICs of ABPC were higher than 2 microg/mL, were included . beta-Lactamase positive ABPC-resistant H . influenzae (BLPAR) were 8 strains (5.2%) . Although those BLNAR were also resistant to variety of beta-lactams, fluoroquinolones and other antibiotics were not affected by the level of ABPC-resistance . Resistant strains of BLPAR against SBT/ABPC, a combination of a beta-lactamase inhibitor, were detected but all of them were sensitive to TAZ/PIPC, an another combination . Those strains were able to be considered as beta-lactamase positive amoxicillin-clavulanate resistant H . influenzae (BLPACR) . PIPC, TAZ/PIPC, CTRX, CDTR, MEPM, LVFX and CPFX showed good activity among tested antibiotics.

Jpn J Antibiot, 2004 Apr, 57(2), 157 - 71
{Antimicrobial susceptibility of Streptococcus pneumoniae, Haemophilus influenzae and Pseudomonas aeruginosa isolated in major hospitals in Nagano prefecture}; Hachiya T et al.; We determined the minimum inhibitory concentration (MIC) of various antimicrobial agents against 140 strains of Streptococcus pneumoniae, 131 strains of Haemophilus influenzae and 178 strains of Pseudomonas aeruginosa isolated from respiratory organs in 28 affiliated hospitals in Nagano prefecture between December 2002 and February 2003 . The results of this report were as followed: 1 . All 140 strains of S . pneumoniae were classified into 3 groups; penicillin-susceptible S . pneumoniae (PSSP) (47.1%), penicillin-intermediate S . pneumoniae (PISP) (43.6%) and penicillin-resistant S . pneumoniae (PRSP) (9.3%) . 2 . Carbapenems and glycopeptide (vancomycin) had highly potent antimicrobial activity against PISP and PRSP like PSSP . However, some of PISP or PRSP isolates were resistant to cephalosporins and a fluoroquinolone (levofloxacin) . 3 . All 131 strains ofH . Influenzae were also classified into three groups; ampicillin sensitive H . influenzae (73.3%), beta-lactamase producing ampicillin resistant H . influenzae (BLPAR) (8.4%) and beta-lactamase negative ampicillin resistant H . influenzae (BLNAR) (18.3%) . 4 . Carbapenems and a fluoroquinolone had highly potent antimicrobial activity against BLPAR and BLNAR . However, there were clear differences among 4 carbapenems for the antimicrobial activity . Ceftriaxone (CTRX) was the most active among cepharosporins in this study . 5 . The rate of P . aeruginosa isolates resistant to carbapenems, a fluoroquinolone and aminoglycosides were about 11 to approximately 16%, 15% and 0.6 to approximately 8%, respectively . None of the strains was resistant to all 3 antimicrobial classes, but 3 strains were resistant to combination of 2 classes . 6 . The MIC50 and MIC90 values of various antibiotics against S . pneumoniae, H . influenzae and P . aeruginosa were different in all 4 regions . In conclusion, the antimicrobial surveillance programs are important for guiding empiric therapy and for focusing interventional control of antimicrobial resistance in regions and hospitals.

Przegl Epidemiol, 2004, 58(1), 57 - 65
{Meningitis and encephalitis in Poland in 2002}; Rosinska M et al.; In Poland, 861 cases of bacterial meningitis and encephalitis, 307 cases of viral encephalitis, 1022 of viral meningitis and 286 cases of meningitis and encephalitis of other or unknown etiology were reported in 2002 . Incidence of the bacterial central nervous system infections has been declining over the past decade and the level of viral infections, following the outbreak in the mid-nineties, remained stable . Etiological factor was determined in 365 (42%) cases of bacterial meningitis/encephalitis . Among them Neisseria meningitidis was found in 90 cases, Haemophilus influenzae in 72 cases and Streptococcus pneumoniae in 85 cases . As in the past type B was the predominant type of N . meningitidis cultured from the patients, but type C appears to be on the rise accounting for 35% of the serotyped strains . 126 cases of tick borne encephalitis were reported in Poland in 2002, most of them from endemic areas of north-eastern part of the country.

J Antibiot (Tokyo), 2004 Apr, 57(4), 280 - 8
In vitro microbiological characterization of a novel azalide, two triamilides and an azalide ketal against bovine and porcine respiratory pathogens; Norcia LJ et al.; Several novel 15-membered-ring macrolide agents (azalide 1, triamilides 2 and 3, and the azalide 3,6-ketal 4) were identified as potential antibacterial agents against Mannheimia (formerly named as Pasteurella) haemolytica, Pasteurella multocida, Haemophilus somnus and Actinobacillus pleuropneumoniae, important etiological agents of bovine and porcine respiratory disease . Compound 3 is the major component of the antibiotic tulathromycin . Antibacterial activity against tilmicosin-resistant P . multocida field isolates was also tested . In vitro MIC 50/90 analysis revealed that the four newly synthesized compounds were more potent than tilmicosin against M . haemolytica (4 to approximately 8x), P . multocida (8 to approximately 16x), A . pleuropneumoniae (4x), H . somnus (2x and 16x), and tilmicosin-resistant P . multocida (32x) . In time-kill kinetic studies, all four novel compounds and tilmicosin showed bactericidal activity against M . haemolytica, P . multocida and A . pleuropneumoniae at both 4x and 8x MIC . A functional assay using genetically defined mutants revealed that all four novel compounds were poorer substrates for the efflux pump, AcrA/B system, than tilmicosin . A pH study using LPS mutants indicated that the enhanced in vitro potency of the triamilides, particularly compound 3 was mainly due to better penetration of the molecule through the outer membrane . The third amine group at the C-4'' position of the triamilde molecules contributed to this increased membrane penetration by increasing overall basicity . These studies indicate that the four novel compounds have potential as antibacterial agents against bovine and porcine respiratory disease.

J Antimicrob Chemother, 2004 Aug, 54(2), 542 - 5 Epub 2004 Jun 23.
Selection of resistance of telithromycin against Haemophilus influenzae, Moraxella catarrhalis and streptococci in comparison with macrolides; Drago L et al.; OBJECTIVE: The in vitro abilities of telithromycin, azithromycin and clarithromycin to select for resistance were compared by testing isolates of Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pneumoniae and beta-haemolytic streptococci . METHODS: Five strains each of beta-lactamase-positive and beta-lactamase-negative H . influenzae, beta-lactamase-positive and beta-lactamase-negative M . catarrhalis, S . pneumoniae, beta-haemolytic group A, group C and group G streptococci and three strains of beta-lactamase-negative ampicillin-resistant H . influenzae were evaluated . Development of resistance was determined by multi-step and single-step methodologies . For multi-step studies, MIC values were determined after five serial passages on antibiotic-gradient plates and after 10 serial passages on antibiotic-free plates . Acquisition of resistance was defined as an increase of >/=4-fold from the starting MIC . In single-step studies, the rate of spontaneous mutations was calculated after a passage on plates containing antibiotics at concentrations equal to the highest NCCLS breakpoints . RESULTS: Azithromycin, clarithromycin and telithromycin gave a >/=4-fold increase in 20, 20 and 10 streptococcus strains, in 4, 5 and 0 H . influenzae strains and in 2, 7 and 4 M . catarrhalis strains, respectively . After 10 passages on antibiotic-free plates, 21/26 strains for azithromycin, 22/32 for clarithromycin and 1/14 for telithromycin maintained high MIC values . In single-step studies, the frequency of mutations was <10(-10) for H . influenzae and M . catarrhalis for telithromycin, azithromycin and clarithromycin . Telithromycin induced mutations at a lower rate than azithromycin and clarithromycin in streptococcal strains . CONCLUSION: Telithromycin showed a very limited ability to select for resistance in respiratory pathogens compared with azithromycin and clarithromycin.

Antimicrob Agents Chemother, 2004 Jul, 48(7), 2599 - 603
Antibacterial effects of amoxicillin-clavulanate against Streptococcus pneumoniae and Haemophilus influenzae strains for which MICs are high, in an in vitro pharmacokinetic model; MacGowan AP et al.; The antibacterial effect of amoxicillin-clavulanate in two formulations, pharmacokinetically enhanced 16:1 amoxicillin-clavulanate twice a day (b.i.d.) and standard 7:1 amoxicillin-clavulanate b.i.d., were studied in an in vitro pharmacokinetic model of infection . Five strains of Streptococcus pneumoniae and two of Haemophilus influenzae, all associated with raised MICs (2 to 8 mg/liter), were used . The antibacterial effect was measured over 24 h by the area under the bacterial kill curve (AUBKC) and the log change in viable count at 24 h (Delta24) . A high 10(8) CFU/ml and low 10(6) CFU/ml initial inocula were used . Employing the Delta24 effect measure, the time above MIC (T>MIC) 50% maximum effect (EC(50)) for S . pneumoniae was in the range 21 to 28% with an 80% maximal response of 41 to 51%, for the AUBKC measure, the value was 26 to 39%, irrespective of inoculum . For H . influenzae, the T>MIC EC(50) was 28 to 37%, and the 80% maximum response was 32 to 48% for the Delta24 measure and 20 to 48% for AUBKC . The maximum response occurred at a T>MIC of 50 to 60% for both species and inocula . The S . pneumoniae data were analyzed by analysis of variance to assess the effect of inoculum, formulation, and MIC on antibacterial effect . Standard and enhanced formulations had different effects depending on MIC, with the standard formulation less effective at higher amoxicillin-clavulanate MICs . This is explained by the greater T>MICs of the enhanced formulation . Although resistant to amoxicillin-clavulanate by conventional breakpoints, S . pneumoniae and H . influenzae strains for which MICs are 2 or 4 mg/liter may well respond to therapy with pharmacokinetically enhanced formulation amoxicillin-clavulanate.

Drugs, 2004, 64(13), 1433 - 64
Cefdinir: a review of its use in the management of mild-to-moderate bacterial infections; Perry CM et al.; Cefdinir (Omnicef) is an oral third-generation cephalosporin with good in vitro activity against many pathogens commonly causative in community-acquired infections . The drug provides good coverage against Haemophilus influenzae, Moraxella catarrhalis and penicillin-susceptible Streptococcus pneumoniae, the most common respiratory tract pathogens . Cefdinir is stable to hydrolysis by commonly occurring plasmid-mediated beta-lactamases and retains good activity against beta-lactamase-producing strains of H . influenzae and M . catarrhalis . The drug distributes into various tissues (e.g . sinus and tonsil) and fluids (e.g . middle ear), and has a pharmacokinetic profile that allows for once- or twice-daily administration.Cefdinir, administered for 5 or 10 days, has shown good clinical and bacteriological efficacy in the treatment of a wide range of mild-to-moderate infections of the respiratory tract and skin in adults, adolescents and paediatric patients in randomised, controlled trials . In adults and adolescents, cefdinir is an effective treatment for both lower (acute bacterial exacerbations of chronic bronchitis {ABECB}, community-acquired pneumonia) and upper (acute bacterial rhinosinusitis, streptococcal pharyngitis) respiratory tract infections, and uncomplicated skin infections . Its bacteriological and clinical efficacy in patients with lower respiratory tract infections was equivalent to that of comparator agents (cefprozil {bacteriological only}, loracarbef, cefuroxime axetil and cefaclor) . In one trial in patients with ABECB, cefdinir produced a higher rate of clinical cure than cefprozil (95% CIs indicated nonequivalence) . Cefdinir also produced good clinical and bacteriological responses equivalent to responses with amoxicillin/clavulanic acid in patients with acute bacterial rhinosinusitis . In addition, it was at least as effective as penicillin V (phenoxymethylpenicillin) in streptococcal pharyngitis/tonsillitis and as effective as cefalexin in uncomplicated skin infections . In paediatric patients aged > or =6 months, cefdinir showed similar efficacy to that of amoxicillin/clavulanic acid or cefprozil in acute otitis media, and cefalexin in uncomplicated skin infections . Cefdinir given for 5 or 10 days was at least as effective as penicillin V for 10 days in patients with streptococcal pharyngitis/tonsillitis . Cefdinir is usually well tolerated . Diarrhoea was the most common adverse event in trials in all age groups . Although the incidence of diarrhoea in cefdinir recipients was generally higher than in adults and adolescents treated with comparators, discontinuation rates due to adverse events were generally similar for cefdinir and comparator groups.In conclusion, cefdinir is a third-generation cephalosporin with a broad spectrum of antibacterial activity encompassing pathogens that are commonly causative in infections of the respiratory tract or skin and skin structure . Depending on the infection being treated, cefdinir can be administered as a convenient once- or twice-daily 5- or 10-day regimen . Clinical evidence indicates that cefdinir is an effective and generally well tolerated drug with superior taste over comparator antibacterial agents and is therefore a good option for the treatment of adults, adolescents and paediatric patients with specific mild-to-moderate respiratory tract or skin infections, particularly in areas where beta-lactamase-mediated resistance among common community-acquired pathogens is a concern.

Pediatr Pulmonol, 2004 Aug, 38(2), 135 - 9
Antigen detection for the diagnosis of pneumonia; Nunes AA et al.; Streptococcus pneumoniae and Haemophilus influenzae type b are the main agents of bacterial community-acquired pneumonia in developing countries, although a definite etiologic diagnosis cannot be established in most cases . This study was carried out to assess the performance of a latex particle agglutination test (LPAT) from a commercial kit (Slidex Meningite Kit trade mark, BioMerieux, France) in diagnosing pneumococcal and H . influenzae type b pneumonia . One hundred and seven children (45 ill subjects and 62 healthy controls) were enrolled . All 45 cases had a presumptive diagnosis of bacterial pneumonia based on clinical (WHO criteria), laboratory (white blood cell count > or = 15.000/mm3, polymorphonuclear leukocytes > or = 70%, bands > or = 500/mm3, and C-reactive protein > or = 40 mg/l), and radiological findings, i.e., two or more positive points in the scoring system described by Khamapirad and Glezen (Semin Respir Infect 1987;2:130-144) . Clinical, laboratory, and radiological assessments were performed in a blinded manner . LPAT was performed in urine samples after concentration through an ethanol-acetone solution . Sensitivity, specificity, and positive and negative predictive values were 77.3% (95% CI, 61.8-88.0%), 90.3% (95% CI, 79.5-96.0%), 85.0% (95% CI, 69.5-93.8%), and 84.8% (95% CI, 73.4-92.1%), respectively . Results suggest that LPAT is a useful diagnostic tool for the etiologic diagnosis of S . pneumoniae and H . influenzae type b pneumonia, especially in the developing world .

J Bacteriol, 2004 Jul, 186(13), 4407 - 11
Haemophilus somnus possesses two systems for acquisition of transferrin-bound iron; Ekins A et al.; Haemophilus somnus strain 649 was found to acquire iron from ovine, bovine, and goat transferrins (Tfs) . Expression of Tf receptors, as evaluated by solid-phase binding assays, required the organisms to be grown under iron-restricted conditions in the presence of Tf . Competition binding assays revealed the presence of two distinct Tf-binding receptor systems, one specific for bovine Tf and the other capable of binding all three ruminant Tfs . Affinity isolation procedures using total membranes yielded three putative bovine Tf-binding polypeptides and one putative ovine and goat Tf-binding polypeptide . PCR amplification followed by DNA sequence analyses revealed that H . somnus strain 649 possesses genes that encode a bipartite TbpA-TbpB receptor along with a homolog of the Histophilus ovis single-component TbpA receptor . Expression of TbpB and the single-component TbpA would appear to be subject to a form of phase variation involving homopolymeric nucleotide tracts within the structural genes.

J Bacteriol, 2004 Jul, 186(13), 4209 - 17
Evolutionary and functional relationships among the nontypeable Haemophilus influenzae HMW family of adhesins; Buscher AZ et al.; Nontypeable Haemophilus influenzae (NTHi) is a common cause of localized respiratory tract disease and initiates infection by colonizing the nasopharynx . Approximately 75 to 80% of NTHi clinical isolates produce proteins that belong to the HMW family of adhesins, which are believed to facilitate colonization . The prototype HMW adhesins are designated HMW1 and HMW2 and were identified in NTHi strain 12 . HMW1 and HMW2 are 71% identical and 80% similar overall, yet display differing cellular binding specificities . In the present study we set out to define more clearly the relationships between HMW1 and HMW2 and other members of the HMW family of adhesins . PCR analysis of 49 epidemiologically distinct isolates revealed that all strains possessing hmw genes as determined by Southern analysis contain two hmw loci in conserved, unlinked physical locations on the chromosome . Functional analysis of the HMW adhesins produced by three unrelated strains demonstrated that each isolate possesses one protein with HMW1-like adherence properties and another with HMW2-like adherence properties . These findings suggest that the hmw1 and hmw2 loci may have arisen via a gene duplication event in an ancestral strain . In addition, they support the hypothesis that the distinct binding specificities of HMW1 and HMW2 emerged early and have persisted over time, suggesting an ongoing selective advantage.

Scand J Infect Dis, 2004, 36(4), 269 - 73
Diagnosis of atypical pathogens in patients hospitalized with community-acquired respiratory infection; Schneeberger PM et al.; The object of our study was to determine the proportion of atypical respiratory pathogens among patients hospitalized with a community-acquired respiratory infection . From September 1997 to May 1999, 159 patients (57% male, median age 55, range 1-88 y) admitted to 3 regional hospitals for a community acquired respiratory infection, were enrolled in the study . Microbiological diagnosis for the atypical pathogens Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella pneumophila was performed with PCR on a throat swab, sputum and/or broncho alveolar lavage (BAL) . In addition, Legionella species other than L . pneumophila (L . non-pneumophila species) were detected by PCR . Two serum samples were collected and processed for M . pneumoniae and C . pneumoniae serology . In total, 27 patients (17%) were diagnosed with an atypical pathogen . Infection with M . pneumoniae was detected in 19 patients (12%) (PCR positive n = 7), with C . pneumoniae in 5 patients (3%) (PCR positive n = 0) and with L . pneumophila in 4 patients (2.5%) (PCR positive n = 4) . In 54 (34%) patients routine microbiological investigations revealed aetiological agents other than the 3 atypical pathogens, the most frequently diagnosed pathogens being Streptococcus pneumoniae (n = 18), Haemophilus influenzae (n = 17), Gram-negative rods (n = 13), Moraxella catarrhalis (n = 6) and Staphylococcus aureus (n = 6) . More than 1 pathogen was found in 13 patients . Atypical pathogens were found more often in the young age group (0-18 y), in contrast to bacterial pathogens that were found more often in the older age groups (> or = 65 y) . Atypical pathogens were found less often in patients with a clinical presentation of atypical pneumonia . Legionella species other than L . pneumophila were found by PCR in 13 patients (8%), and in 6 patients in combination with another pathogen . An atypical pathogen (M . pneumoniae, C . pneumoniae or L . pneumophila) was found in 17% of the patients hospitalized with a community acquired respiratory infection, predominantly in the young age group . The role of Legionella non-pneumophila species as pathogen in community acquired respiratory infection needs to be determined . The clinical presentation does not predict the type of pathogen found.

Int J Antimicrob Agents, 2004 Jun, 23(6), 533 - 46
Antimicrobial selection for community-acquired lower respiratory tract infections in the 21st century: a review of gemifloxacin; Appelbaum PC et al.; Community-acquired lower respiratory tract infections (LRTIs) are more prevalent in the elderly than in children and younger adults and form a significant proportion of all consultations and hospital admissions in this older age group . Furthermore, in a world of increasing life expectancy the trend seems unlikely to be reversed . Antimicrobial treatment of community-acquired pneumonia (CAP) must cover Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, and in many circumstances should also cover the intracellular (atypical) pathogens . In contrast, acute exacerbations of chronic bronchitis (AECB) are mainly associated with H . influenzae and S . pneumoniae and not with atypical bacteria: in severe cases, other Gram-negative bacteria may be involved . Frequently in LRTIs, the aetiology of the infection cannot be identified from the laboratory specimens and treatment has to be empirical . In such situations it is important to not only to use an antibiotic that covers all likely organisms, but also one that has good activity against these organisms given the local resistance patterns . Gemifloxacin is a new quinolone antibiotic that targets pneumococcal DNA gyrase and topoisomerase IV and is highly active against S . pneumoniae including penicillin-, macrolide- and many ciprofloxacin-resistant strains, as well as H . influenzae and the atypical pathogens . In clinical trials in CAP and AECB, gemifloxacin has been shown to be as effective a range of comparators and demonstrated an adverse event profile that was in line with the comparator agents . In one long-term study in AECB significantly more patients receiving gemifloxacin than clarithromycin remained free of recurrence after 26 weeks . The improved potency, broad spectrum of activity and proven clinical and bacteriological efficacy and safety profile should make it a useful agent in the 21st century battle against community-acquired LRTIs .

Clin Microbiol Infect, 2004 Jun, 10(6), 521 - 6
Antimicrobial susceptibility to levofloxacin and other antibacterial agents among common respiratory pathogens-a Brazilian perspective from the GLOBAL Surveillance Initiative 2001-2002; Mendes C et al.; The GLOBAL (Global Landscape On Bactericidal Activity of Levofloxacin) Surveillance programme monitored antimicrobial susceptibility patterns of the key respiratory tract pathogens Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis collected in Brazil during 1997-1998, 1999-2000 and 2001-2002 . Penicillin and azithromycin resistance among S . pneumoniae strains increased from 1997-1998, reaching 7.9% and 9.5%, respectively, in 2001-2002 . Although decreasing by 4.9% since the previous study, trimethoprim-sulphamethoxazole resistance remained high at 33.7% . Concurrent resistance to penicillin, azithromycin and trimethoprim-sulphamethoxazole was seen in 2.9% of the S . pneumoniae isolates collected . Levofloxacin remained extremely active against S . pneumoniae, with 0.3% resistance reported in 1997-1998 and 0% resistance in 1999-2000 and 2001-2002 . beta-Lactamase production in H . influenzae was > 10% in all three studies, with correspondingly high rates of ampicillin resistance . Trimethoprim-sulphamethoxazole was the least active agent tested against H . influenzae, with resistance rates of > 40% recorded in all three studies . All H . influenzae isolates were susceptible to cefuroxime, ceftriaxone, azithromycin and levofloxacin . Of the M . catarrhalis isolates, 98.0% in 1997-1998, 98.0% in 1999-2000 and 81.8% in 2001-2002 were beta-lactamase-positive . The continued high prevalence of antimicrobial resistance in Brazil underscores the importance of current surveillance initiatives . Levofloxacin, a fluoroquinolone prescribed widely for respiratory tract infections, continued to show potent activity against key respiratory pathogens.

J Antimicrob Chemother, 2004 Jul, 54(1), 225 - 8 Epub 2004 Jun 09.
Ultrastructure of Streptococcus pneumoniae after exposure to xylitol; Tapiainen T et al.; OBJECTIVES: Xylitol is a sugar alcohol which reduces the growth of Streptococcus pneumoniae and the adherence of pneumococci and Haemophilus influenzae to nasopharyngeal cells . Xylitol prevents acute otitis media but does not decrease nasopharyngeal carriage of pneumococci . We hypothesized that xylitol could affect the surface structures of viable pneumococci, which would further explain the mechanism of action of xylitol in preventing acute otitis media . METHODS: We exposed five strains of pneumococci to 0.5%-5% xylitol, 5% glucose, 5% fructose and 5% sorbitol or control medium (brain heart infusion) for 0.5-2 h and examined the ultrastructure of bacteria by electron microscopy . RESULTS: The cell wall of pneumococci became more diffuse, the polysaccharide capsule became ragged and the proportion of damaged pneumococci increased after exposure to xylitol for 2 h, but not after exposure to other sugars or control medium . The phenotype of all pneumococcal strains was opaque before xylitol exposure and became almost transparent both in xylitol and in control medium during the experiment . CONCLUSIONS: This study demonstrates further that xylitol has a harmful effect on pneumococci . The observed changes in the polysaccharide capsule and the cell wall of pneumococci could affect the adherence and virulence of pneumococci, explaining the good clinical efficacy of xylitol in the prevention of acute otitis media.

Clin Ther, 2004 Apr, 26(4), 522 - 30
Comparative in vitro activity of telithromycin and beta-lactam antimicrobials against community-acquired bacterial respiratory tract pathogens in the United States: findings from the PROTEKT US study, 2000-2001; Stratton CW et al.; BACKGROUND: Telithromycin is a new ketolide antimicrobial that was developed to provide good activity against resistant respiratory tract pathogens . PROTEKT US (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin in the United States) is a multicenter in vitro surveillance study that was initiated in 2000 to chart the emergence and spread of antimicrobial-resistant pathogens in patients with community-acquired respiratory tract infections (CARTIs) . OBJECTIVE: This article reports first-year results from PROTEKT US pertaining to the comparative in vitro activity of telithromycin and beta-lactam antimicrobials against community-acquired bacterial respiratory tract pathogens . METHODS: Data were compiled on the comparative in vitro activity of telithromycin and beta-lactams against Streptococcus pneumoniae (10,103 isolates), Streptococcus pyogenes (3918 isolates), and Haemophilus influenzae (2706 isolates) . Minimum inhibitory concentrations (MICs) and susceptibilities were determined according to National Committee for Clinical Laboratory Standards methods . RESULTS: In total, 38.8% (3920/10,103) of pneumococcal isolates were not susceptible to penicillin (12.5% {1266} intermediate {MIC, > or =0.12-1.0 mg/dL}, 26.3% {2654} resistant {MIC, > or =2 mg/dL}) . Telithromycin was highly active against S pneumoniae (MIC required to inhibit 90% of isolates {MIC(90)}, 0.5 mg/L), with 99.6% (10,062/10,103) of isolates fully susceptible (MIC, < or =1 mg/L) . Based on MIC(90)s, the rank order of antimicrobial activity was telithromycin (0.5 mg/L), followed by amoxicillin/clavulanate (2 mg/L), penicillin (4 mg/L), and cefuroxime (8 mg/L) . Telithromycin retained high activity (MIC(90), 1 mg/L) against penicillin-resistant penumococci that showed high levels of coresistance to beta-lactams . All isolates of S pyogenes were fully susceptible to the beta-lactams tested . Beta-lactamase production was common among H influenzae isolates (28.3% {765/2706}) . Telithromycin was active against H influenzae (MIC(90), 4 mg/L), irrespective of beta-lactamase production . Conclusion: Overall, these findings from the first year of PROTEKT US support the potential value of telithromycin in the treatment of CARTIs.

Ann Pharmacother, 2004 Jul-Aug, 38(7-8), 1226 - 35 Epub 2004 Jun 08.
Gemifloxacin: a new fluoroquinolone approved for treatment of respiratory infections; Yoo BK et al.; OBJECTIVE: To evaluate the microbiology, pharmacokinetic parameters, drug interactions, and results of the available clinical trials of gemifloxacin for the treatment of community-acquired pneumonia (CAP) and acute exacerbation of chronic bronchitis (AECB) . DATA SOURCES: MEDLINE (1966-September 2003) was searched for primary and review articles . Data from the manufacturer were also included . Key words included adverse effects, clinical trials, drug interactions, gemifloxacin, and pharmacokinetic parameters . STUDY SELECTION AND DATA EXTRACTION: All articles and product labeling concerning gemifloxacin, a fluoroquinolone antibiotic recently approved by the Food and Drug Administration for treatment of CAP and AECB, were included for review . DATA SYNTHESIS: Compared with currently available fluoroquinolones, gemifloxacin demonstrated improved in vitro activity against Streptococcus pneumoniae (minimum inhibitory concentration for 90% eradication 0.03 microg/mL) and similar activity against gram-negative respiratory pathogens (Haemophilus influenzae, Moraxella catarrhalis) and atypical pathogens such as Chlamydia pneumoniae, Legionella pneumophila, and Mycoplasma pneumoniae . Gemifloxacin, consistent with other available fluoroquinolones, has insufficient activity against methicillin-resistant Staphylococcus aureus to allow clinical use for such infections . Gemifloxacin has adequate bioavailability and a favorable drug interaction profile . Gemifloxacin was comparable to commonly employed nonfluoroquinolone regimens for treatment of CAP and AECB, although the studies were designed to demonstrate equivalence . Gemifloxacin once daily for 5-7 days was well tolerated in controlled and uncontrolled clinical studies . Available clinical data, however, are insufficient to draw clinical or toxicologic distinctions between gemifloxacin and other fluoroquinolones . CONCLUSIONS: Gemifloxacin may be a suitable choice for empiric treatment of CAP or AECB . However, due to the significant history of fluoroquinolone-induced hepatic failure and dermatologic complications, the use of this drug should be closely monitored.

Int J STD AIDS, 2004 Jun, 15(6), 367 - 70
Genital syndromes and syndromic management of vaginal discharge in a community setting; George R et al.; The objective of the study was to determine the community prevalence of genital syndromes in women and evaluate the syndromic management of vaginal discharge in this setting . A representative sample for the state of Tamilnadu was chosen using probability proportional to size cluster technique . Thirty clusters were selected from three districts . Demographic, sexual behaviour, risk factors, clinical and laboratory data were collected from the selected population using a structured questionnaire . Direct smear examination for Trichomonas vaginalis, culture for Neisseria gonorrhoeae and Haemophilus ducreyi, serological tests for syphilis (RPR and TPHA), hepatitis B (Hbs Ag ELISA), IgM and IgG antibodies to HSV2 (Novum diagnostics, Germany) and PCR test for detection of C . trachomatis from urine were done . There were 1157 women in the selected population . On examination, vaginal discharge was the most common genital syndrome (38.4%) . The sensitivity, specificity, positive and negative predictive value of vaginal discharge as a marker for STD in women was found to be 43.3%, 61.6%, 10.7% and 91.1%, respectively . We concluded that treatment on the basis of syndromic management would result in over-treatment of 90% of women with vaginal discharge.

AIDS Res Hum Retroviruses, 2004 May, 20(5), 493 - 6
Haemophilus influenzae type b immunization in adults infected with the human immunodeficiency virus; De Sousa dos Santos S et al.; The purpose of this study was to assess the influence of Haemophilus influenzae type b conjugate vaccine on HIV-1 RNA level, CD4 count, and anti-Hib polysaccharide (PRP) antibody concentration . Eighty HIV-infected adults were randomized to receive Hib conjugate vaccine or not . Twenty HIV-seronegative controls were also vaccinated . Blood samples were taken before and after vaccination, with a follow-up period of 6 months . HIV infection markers and anti-PRP antibodies were monitored . There was no change in either HIV-1 viremia or CD4 count after vaccination . Immunization immunogenicity was superior in HIV-uninfected than in HIV-infected individuals (p < 0.01) . Hib vaccination was safe but induced suboptimal antibody response in HIV-infected adults.

Pediatr Dent, 2004 May-Jun, 26(3), 283 - 8
Comparison of oral findings in special needs children with and without gastrostomy; Jawadi AH et al.; PURPOSE: The objective of this study was to compare aspiration pneumonia (AP)-associated microflora, calculus, and oral hygiene/care seeking behaviors in special health care needs children (SC) with gastrostomy (GT) and without . METHODS: Twenty-seven GT SC, ages 3 to 12 years old and matched for age and gender with 27 non-GT SC, were examined by 2 trained investigators who recorded calculus and gingival inflammation and reconciled differences . Plaque was obtained using preweighed dry paper points and saliva sampled using sterile pipettes and cultured using standard bioassay procedures in a hospital laboratory . Parent/caretakers completed a medical and oral health questionnaire . RESULTS: No significant differences were noted for age, gender, weight, primary diagnosis, vomiting, constipation, or swallowing disorder, but GT children received 4 medications vs 1 for non-GT and were significantly more likely to have had AP, need special feeding, and drool (P < or = .05) . Oral health measures were not significantly different for brushing frequency, dentifrice use, brushing problems, frequency of dental care, or gingival inflammation, but GT patients had significantly more plaque and calculus . GT patients had significantly more Haemophilus influenzae, with trends toward more gram negative enteric rods, pseudomonas, and Streptococcus pneumoniae and higher concentrations in several GT-patients and little or none in non-GT patients . GT SC had significantly less beta-streptococci than non-GT patients (P < or = .05) . CONCLUSIONS: GT SC had significantly more of 1 AP-associated organism than non-GT SC and significantly more calculus and plaque, in spite of similar care seeking and hygiene behaviors.

Epidemiol Mikrobiol Imunol, 2004, 53(2), 74 - 7
{Characterization of Haemophilus influenzae strains using multilocus sequencing}; Krizova P et al.; First results of multilocus sequence typing (MLST) of Haemophilus influenzae strains are presented . MLST of 28 H . influenzae strains isolated from patients with invasive diseases in the Czech Republic is indicative of clonal homogeneity of these strains: 22 out of 26 H . influenzae b strains tested were of the same sequence type, ST-6 . Four strains were of two sequence types newly described in this study: ST-83 (3 strains) and ST-84 (1 strain) . Two nontypeable H . influenzae strains were assigned to sequence types other than ST-6: ST-3 and ST-85 newly described in this study . First MLST results show ST-6 to be typical of H . influenzae b isolated from patients with invasive diseases in the Czech Republic . The sequence types newly described in this study, i.e . ST-83, ST-84 and ST-85, were submitted to the worldwide H . influenzae MLST database .

Onderstepoort J Vet Res, 2004 Mar, 71(1), 53 - 8
Evidence of possible evasion of protective immunity by NAD-independent isolates of Haemophilus paragallinarum in poultry; Bragg RR; An indication of the ability of NAD-independent variants of Haemophilus paragallinarum to evade the immune system has been obtained from data obtained from several experiments . Firstly, it was noted that there was a difference in the serovar distribution between the NAD-dependent isolates in the 1990s and the NAD-independent isolates, as there was a significant decrease in the incidence of serogroup A NAD-dependent isolates . This can possibly be attributed to the extensive use of vaccines . On the other hand, most of the earlier NAD-independent isolates were serovar A . This is a possible indication of evasion of the protective immunity by the NAD-independent isolates . Further evidence of possible evasion of the protective immunity was obtained from results obtained when different isolates, both NAD dependent and NAD independent, were tested with a panel of monocional antibodies (Mabs) . The V1 Mab reaction pattern was only seen in the reference strain 0083 among all of the NAD-dependent isolates tested in South Africa . This Mab was, however, found to react with some of the NAD-independent isolates . Furthermore, the isolation of NAD-dependent isolates in Australia which react with the V1 Mab also suggest possible evasion of the protective immunity by the NAD-independent isolates as no vaccines containing strain 0083 are used in Australia . In order to investigate the hypothesis of immune-evasion by NAD-independent H . paragallinarum, vaccinated and unvaccinated chickens were challenged with a NAD-independent serogroup C isolate . As a control, chickens were also challenged with NAD-dependent H . paragallinarum of the same serogroup . The results obtained indicate that there is no significnat difference in the disease profiles obtained in vaccinated and unvaccinated chickens challenged with the NAD-independent isolate, thus providing further evidence of evasion of the productivity immunity by the NAD-independent isolates . The ability of the NAD-independent isolates to evade the immune system suggests that a different vaccination strategy, or alternative control methods may be needed for the control of IC caused by these isolates.

Onderstepoort J Vet Res, 2004 Mar, 71(1), 1 - 8
Limitation of the spread and impact of infectious coryza through the use of a continuous disinfection programme; Bragg RR; The effect of a continuous disinfection programme, using the non-toxic disinfectant Virukill, in layers, on the spread and impact of infectious coryza, caused by Haemophilus paragallinarum was evaluated . In this experiment, both unvaccinated layers and layers vaccinated against infectious coryza were used . Duplicate smaller groups of vaccinated and unvaccinated chickens were challenged with different serovars of both NAD-dependent as well as NAD-independent isolates of Haemophilus paragallinarum . One group of chickens challenged with each of the different becterial serovars was treated with the continuous disinfection programme, while the other group remained as the untreated controls . The clinical signs of infectious coryza were evaluated over a period of 20 days in each group . The egg production over this period was also evaluated . It was found in all experimental challenges, that the severity of the symptoms was reduced in the birds receiving the continuous disinfection programme . The drop in egg production was also found to be less severe in the treated groups when compared to the untreated control groups . The duration of infection was found to be either unchanged, or shorter in the birds treated with the continuous disinfection programme . In none of the experimental challenges was the duration or expression of clinical signs of IC increased due to the continuous disinfection programme.

Respirology, 2004 Jun, 9(2), 278 - 82
Clinical features of Q fever pneumonia; Okimoto N et al.; The aim of the study was to assess the clinical features of Q fever pneumonia in Japan . Four cases of Q fever pneumonia (a female aged 21 and males aged 53, 74 and 87 years) who were diagnosed using the PanBio ELISA test kit, were assessed and their clinical features are described . The frequency of Q fever pneumonia among our cases of community-acquired pneumonia was 1.4% (4/284) . A 21-year-old female had a typical case of the disease with (i) a history of owning a cat, (ii) onset with fever and dry cough, (iii) multiple soft infiltrative shadows on CXR, (iv) a normal white blood cell count, and (v) good response to clarithromycin . The pneumonias in the other three cases were considered mixed infections with bacteria such as Streptococcus pneumoniae and Haemophilus influenzae . Their clinical features included the following: (i) an elderly person with an underlying disease, (ii) onset with fever and purulent sputum, (iii) coarse crackles on auscultation, (iv) infiltrative shadows and pleural effusion on CXR, (v) increased white blood cells with elevated BUN and hyponatraemia, and (vi) modest responses to combined therapy with carbapenem and minocycline . Our observations suggest that two types of pneumonia caused by Coxiella burnetti exist; one with the usual features of atypical pneumonia, and the other presenting with the clinical features of bacterial pneumonia in the elderly due to mixed bacterial infection.

Clin Lab Med, 2004 Jun, 24(2), 531 - 51
Evolution of amoxicillin/clavulanate in the treatment of adults with acute bacterial rhinosinusitis and community-acquired pneumonia in response to antimicrobial-resistance patterns; File TM Jr et al.; Current treatment guidelines for community-acquired respiratory tract infections no longer depend solely on the characteristics of the patient and the clinical syndrome, but on those of the offending pathogen, including presence and level of antimicrobial resistance . The most common respiratory tract pathogens known to cause acute bacterial rhinosinusitis (ABRS) and community-acquired pneumonia (CAP) include Streptococcus pneumoniae and Haemophilus influenzae . The prevalence of antimicrobial resistance, especially b-lactum and macrolide resistance, among S pneumoniae and H influenzae has increased dramatically during the past 2 decades, diminishing the activity of many older antimicrobials against resistant organisms . A pharmacokinetically enhanced formulation of amoxicillin/clavulanate has been developed to fulfill the need for an oral b-lactam antimicrobial that achieves a greater time that the serum drug concentration exceeds the minimum inhibitory concentration (T > MIC) of antimicrobials against pathogens than conventional formulations to improve activity against S pneumoniae with reduced susceptibility to penicillin . The b-lactamase inhibitor clavulanate allows for coverage of b-lactamase-producing pathogens, such as H influenzae and M catarrhalis . This article reviews the rationale for, and evolution of, oral amoxicillin clavulanate for ABRS and CAP

Clin Lab Med, 2004 Jun, 24(2), 503 - 30
Susceptibility of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis to 17 oral antimicrobial agents based on pharmacodynamic parameters: 1998-2001 U S Surveillance Study; Jacobs MR et al.; Pharmacokinetic/pharmacodynamic parameters were used to interpret susceptibility data for the oral agents tested in a clinically meaningful way . Among S pneumoniae isolates, >99% were susceptible to respiratory fluoroquinolones, 91.6% to amoxicillin, 92.1% to amoxicillin/clavulanic acid (95.2% at the extended-release formulation breakpoint), 90.6% to clindamycin, 80.4% to doxycycline, 71.0% to azithromycin, 72.3% to clarithromycin, 71.8% to cefprozil and cefdinir, 72.6% to cefuroxime axetil, 66.3% to cexime, 63.7% to trimethoprim/sulfamethoxazole, and 19.7% to cefaclor . Among H influenzae isolates, 28.6% were b-lactamase positive, but virtually all were susceptible to amoxicillin/clavulanic acid (98.3%, with 99.8% at the extended-release formulation breakpoint), cexime (100%), and uoroquinolones (99.8%), whereas 93.5% were susceptible to cefdinir, 82.8% to cefuroxime axetil, 78.1% to trimethoprim/sulfamethoxazole, 70.2% to amoxicillin, 25.1% to doxycycline, 23.2% to cefprozil, and 5% to cefaclor, azithromycin and clarithromycin . Most isolates of M catarrhalis were resistant to amoxicillin, cefaclor, cefprozil, and trimethoprim/sulfamethoxazole . Thus significant b-lactam and macrolide/azalide resistance in Streptococcus pneumoniae and b-lactamase production and trimethoprim/sulfamethoxazole resistance in untypeable Haemophilus influenzae are still present . The results of this study should therefore be applied to clinical practice based on the clinical presentation of the patient, the probability of the patient's having a bacterial rather than a viral infection, the natural history of the disease, the potential of pathogens to be susceptible to various oral antimicrobial agents, the potential for cross-resistance between agents with S pneumoniae, and the potential for pathogens to develop further resistance . Antibiotics should be used judiciously to maintain remaining activity and chosen carefully based on activity determined by pharmacokinetic/pharmacodynamic-based breakpoints to avoid these bacteria developing further resistance, particularly to fluoroquinolones.

Clin Lab Med, 2004 Jun, 24(2), 455 - 75
Macrolide resistance in Streptococci and Haemophilus influenzae; Bozdogan B et al.; Antimicrobial resistance is a growing problem among pathogens from respiratory tract infections . b-Lactam resistance rates are escalating among Streptococcus pneumoniae and Haemophilus influenzae . Macrolides are increasingly used for the treatment of respiratory tract infections, but their utility is compromised by intrinsic and acquired resistance . This article analyses macrolide-resistance mechanisms and their worldwide distributions in S pneumoniae, S pyogenes, and H influenzae.

Rev Prat, 2004 Mar 15, 54(5), 526 - 31
{Vaccines pharmacovigilance}; Autret-Leca E et al.; The pharmacovigilance of vaccines has the particularity of concerning medications with a preventative target, used in healthy subjects, who are often young . Their individual benefit is deferred and unknown, whereas their risk is immediate . Certain undesirable effects are linked to the antigen of live attenuated vaccines (post-MMR lymphocytic meningitis) . Other non-specific effects are linked to other different components of the vaccines (macrophage and aluminium myofasciitis) . Undesirable events susceptible to being due to the vaccination are identified and managed according to standardised procedures of pharmacovigilance, that is to say, based on "spontaneous notification", generation of an alert, confirmed or not by studies of pharmaco-epidemiology . The studies of pharmaco-epidemiology: have made evident oedematous reactions with cyanosis or purpura, with the vaccines containing the Haemophilus b valence, and the absence of an association with sudden death of the newborn; have excluded the existence of an elevated risk of demyelinisation or auto-immune disease associated with vaccination against hepatitis B, without being able to exclude a slight risk; go against the finding of an association between Crohn's disease and/or autisim and the MMR vaccination . Only their frequently encountered undesirable effects are well identified at the moment of commercialisation . Post-marketing surveillance of vaccines (declaration to the regional pharmacovigilance centres) allow the detection of possible rare and serious effects and the evaluation of the real vaccination risk . Thus it must be intensive and systematic.

Semin Pediatr Infect Dis, 2004 Jan, 15(1), 5 - 20
Antimicrobial resistance among pediatric respiratory tract infections: clinical challenges; Jacobs MR et al.; Considerable development of antimicrobial resistance has occurred in the major pediatric bacterial pathogens, Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis . However, most of the respiratory infections that children suffer are viral and self-limiting, and only a small percentage of them will develop secondary bacterial infections with the pathogens listed . The challenge for rational antibiotic use is to determine which patients can be treated conservatively and which require antimicrobial intervention to avoid prolonged discomfort or development of permanent sequelae . The basis for rational use of antibiotic in the era of resistance in these major pathogens is to avoid overuse of antimicrobial agents, tailor treatment to identified pathogens as much as possible, and base empiric treatment on the disease being treated and the susceptibility of the probable pathogens at breakpoints based on pharmacokinetic and pharmacodynamic parameters . With appropriate dosing regimens based on these parameters and despite development of resistance, amoxicillin is still one of the most active oral agents against S . pneumoniae and non-beta-lactamase producing strains of H . influenzae, whereas amoxicillin-clavulanate is active against beta-lactamase-producing strains of H . influenzae and M . catarrhalis . Parenteral ceftriaxone and oral and parenteral fluoroquinolones are active against all 3 species, but fluoroquinolones should be used with utmost caution when all other options have been considered because of concerns about toxicity and development of resistance . Introduction of a 7-valent conjugate pneumococcal vaccine in the United States in 2000 reduced the prevalence of invasive pneumococcal disease in children younger than 2 years old, but, as of 2001, had not had a major impact on decreasing antimicrobial resistance.

Rev Argent Microbiol, 2004 Jan-Mar, 36(1), 20 - 3
{Isolation of Haemophilus influenzae serotypes from deep sites in sick children}; Gatti BM et al.; Haemophilus influenzae (Hi) is the causative agent of several human diseases such as sepsis, meningitis, celulitis, and osteoarthritis . We investigated the isolation of Hi serotypes from sterile sites in sick children . One hundred and seventy nine strains from 146 patients were studied, period 1996-2002, at the Microbiology Laboratory, Hospital de Ninos Superiora Sor Maria Ludovica, Argentina . The serotype distribution was:1 a, 112 b,1 c,1 d, 4 e, 3 f y 24 no typable . Since the beginning of universal Hi b vaccination in 1998, we have observed the fast decrease of serotype b and a relative increase of other serotypes.

Pediatrics, 2004 Jun, 113(6 Suppl), 1959 - 64
Insurance status and vaccination coverage among US preschool children; Santoli JM et al.; BACKGROUND: Insurance status has been shown to have an impact on children's use of preventive and acute health services . The objective of this study was to determine the relationship between insurance status and vaccination coverage among US preschool children aged 19 to 35 months . METHODS: We linked data from 2 national telephone surveys, the National Immunization Survey and the National Survey of Early Childhood Health, conducted during the first half of 2000 . Children were considered up to date (UTD) when they had received at least 4 diphtheria-tetanus-acellular pertussis/diphtheria-tetanus-pertussis vaccines, 3 poliovirus vaccines, 1 MMR vaccine, 3 Haemophilus influenza vaccines, and 3 hepatitis B vaccines at the time the interview was conducted . RESULTS: Among the 735 children in our study sample, 72% were UTD . The vast majority (94%) reported some type of health insurance at the time of the survey . Children with private insurance were more likely to be UTD (80%) than those with public insurance (56%) or no insurance (64%) . In a multivariate analysis that controlled for child's race/ethnicity; household income; maternal age/marital status/educational level; location of usual care; and Special Supplemental Nutrition Program for Women, Infants, and Children participation, insurance was no longer an independent predictor of vaccination . CONCLUSIONS: The disparity in vaccination coverage among publicly, privately, and uninsured children is dramatic, underscoring its importance as a marker for underimmunization, despite the multivariate findings . The Vaccines for Children Program, a partnership between public health and vaccination providers who serve uninsured children and those enrolled in Medicaid, is well suited to target and improve vaccination coverage among these vulnerable children.

JAMA, 2004 Jun 2, 291(21), 2555 - 62
Monthly antibiotic chemoprophylaxis and incidence of sexually transmitted infections and HIV-1 infection in Kenyan sex workers: a randomized controlled trial; Kaul R et al.; CONTEXT: Sexually transmitted infections (STIs) are common in female sex workers (FSWs) and may enhance susceptibility to infection with human immunodeficiency virus type 1 (HIV-1) . OBJECTIVE: To examine regular antibiotic prophylaxis in FSWs as a strategy for reducing the incidence of bacterial STIs and HIV-1 . DESIGN, SETTING, AND PARTICIPANTS: Randomized, double-blind, placebo-controlled trial conducted between 1998-2002 among FSWs in an urban slum area of Nairobi, Kenya . Of 890 FSWs screened, 466 who were seronegative for HIV-1 infection were enrolled and randomly assigned to receive azithromycin (n = 230) or placebo (n = 236) . Groups were well matched at baseline for sexual risk taking and STI rates . INTERVENTION: Monthly oral administration of 1 g of azithromycin or identical placebo, as directly observed therapy . All participants were provided with free condoms, risk-reduction counseling, and STI case management . MAIN OUTCOME MEASURES: The primary study end point was incidence of HIV-1 infection . Secondary end points were the incidence of STIs due to Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Treponema pallidum, and Haemophilus ducreyi, as well as bacterial vaginosis . Analysis of herpes simplex virus type 2 (HSV-2) infection was performed post hoc . RESULTS: Seventy-three percent of participants (n = 341) were followed up for 2 or more years or until they reached an administrative trial end point . Incidence of HIV-1 did not differ between treatment and placebo groups (4% {19 cases per 473 person-years of follow-up} vs 3.2% {16 cases per 495 person-years of follow-up} rate ratio {RR}, 1.2; 95% CI, 0.6-2.5) . Incident HIV-1 infection was associated with preceding infection with N gonorrhoeae (rate ratio {RR}, 4.9; 95% CI, 1.7-14.3) or C trachomatis (RR, 3.0; 95% CI, 1.1-8.9) . There was a reduced incidence in the treatment group of infection with N gonorrhoeae (RR, 0.46; 95% CI, 0.31-0.68), C trachomatis (RR, 0.38; 95% CI, 0.26-0.57), and T vaginalis (RR, 0.56; 95% CI, 0.40-0.78) . The seroprevalence of HSV-2 infection at enrollment was 72.7%, and HSV-2 infection at baseline was independently associated with HIV-1 acquisition (RR, 6.3; 95% CI, 1.5-27.1) . CONCLUSIONS: Despite an association between bacterial STIs and acquisition of HIV-1 infection, the addition of monthly azithromycin prophylaxis to established HIV-1 risk reduction strategies substantially reduced the incidence of STIs but did not reduce the incidence of HIV-1 . Prevalent HSV-2 infection may have been an important cofactor in acquisition of HIV-1.

Vet Microbiol, 2004 Jun 21, 101(2), 143 - 6
Antimicrobial susceptibility of Haemophilus parasuis and Histophilus somni from pigs and cattle in Denmark; Aarestrup FM et al.; A total of 52 Haemophilus parasuis and 80 Histophilus somni isolates were tested for antimicrobial susceptibility by MIC-determinations . None of the isolates were resistant to ampicillin, ceftiofur, ciprofloxacin, erythromycin, florphenicol, penicillin, spectinomycin, tetracycline, tiamulin, or tilmicosin . Two H . parasuis isolates were resistant to trimethoprim + sulfamethoxazole . Six H . parasuis isolates had reduced susceptibility (0.06-0.5 microg/ml) to ciprofloxacin and 10 reduced susceptibility to TMP + sulfamethoxazole (1-2 microg/ml) . This study showed that Danish isolates of H . parasuis and H . somni in general are fully susceptible to antimicrobial agents currently used for treatment of infections with these pathogens .

Am J Physiol Lung Cell Mol Physiol, 2004 Sep, 287(3), L598 - 607 Epub 2004 May 28.
Modulation of airway inflammation and bacterial clearance by epithelial cell ICAM-1; Humlicek AL et al.; Many cell types in the airway express the adhesive glycoprotein for leukocytes intercellular adhesion molecule-1 (ICAM-1) constitutively and/or in response to inflammatory stimuli . In this study, we identified functions of ICAM-1 on airway epithelial cells in defense against infection with Haemophilus influenzae . Initial experiments using a mouse model of airway infection in which the bacterial inoculum was mixed with agar beads that localize inflammation in airways demonstrated that ICAM-1 expression was required for efficient clearance of H . influenzae . Airway epithelial cell ICAM-1 expression required few or no leukocytes, suggesting that epithelial cells could be activated directly by interaction with bacteria . Specific inhibition of ICAM-1 function on epithelial cells by orotracheal injection of blocking antibodies resulted in decreased leukocyte recruitment and H . influenzae clearance in the airway . Inhibition of endothelial cell ICAM-1 resulted in a similar decrease in leukocyte recruitment but did not affect bacterial clearance, indicating that epithelial cell ICAM-1 had an additional contribution to airway defense independent of effects on leukocyte migration . To assess this possibility, we used an in vitro model of neutrophil phagocytosis of bacteria and observed significantly greater engulfment of bacteria by neutrophils adherent to epithelial cells expressing ICAM-1 compared with nonadherent neutrophils . Furthermore, bacterial phagocytosis and killing by neutrophils after interaction with epithelial cells were decreased when a blocking antibody inhibited ICAM-1 function . The results indicate that epithelial cell ICAM-1 participates in neutrophil recruitment into the airway, but its most important role in clearance of H . influenzae may be assistance with neutrophil-dependent bacterial killing.

Int J Antimicrob Agents, 2004 Mar, 23(3), 296 - 9
Antimicrobial susceptibility of Haemophilus influenzae, Haemophilus parainfluenzae and Moraxella catarrhalis isolated from adult patients with respiratory tract infections in four southern European countries . The ARISE project; Soriano F et al.; Over a 7-month period in 2000-2001, 1213 Haemophilus influenzae, 112 Haemophilus parainfluenzae and 142 Moraxella catarrhalis isolates were recovered from adult patients with respiratory tract infections . Patients were from four southern European countries (Spain, Italy, Portugal and Greece) . The antimicrobial susceptibility of the isolates to 11 antibiotics was determined in a central laboratory . The most active drugs on the basis of MICs were levofloxacin, cefditoren, cefotaxime, cefpodoxime and amoxicillin/clavulanate . MICs > or = 2 mg/l for amoxicillin were found in 19.5, 28.6, and 75.4% of H . influenzae, H . parainfluenzae and M . catarrhalis isolates, respectively . Isolates of H . influenzae and H . parainfluenzae with reduced susceptibility or that were fully resistant to amoxicillin/clavulanate, cefuroxime and clarithromycin were detected (0.2-1.8%) as well as M . catarrhalis resistant to clarithromycin (0.7%) . Regular surveys of resistance patterns for antimicrobial agents are necessary.

Int J Antimicrob Agents, 2004 Mar, 23(3), 218 - 25
PK-PD modelling of the effect of cefaclor on four different bacterial strains; de la Pena A et al.; The effect of cefaclor against relevant bacterial strains was studied by employing a combined in vivo pharmacokinetic (PK)-in vitro pharmacodynamic (PD) approach . For this purpose selected isolates of Escherichia coli, Moraxella catarrhalis, Haemophilus influenzae and Streptococcus pneumoniae were exposed in vitro to the interstitial cefaclor profile obtained in vivo in the interstitial space fluid of human tissue after administration of commonly used doses of cefaclor and the change in the number of colony forming units per millilitre (CFU/ml) versus time was monitored . Fitting of the data using a modified E(max)-model resulted in a set of mean pharmacodynamic parameters (k0, k(max), EC50) for each bacterial strain . The parameters derived from these experiments were used in a computer-simulation of the antibacterial effects for different dosing regimens and formulations of cefaclor, notably an immediate (IR) and a modified (MR) release formulation . Dosage regimens were compared using the ratio between the number of bacteria remaining after 24 h of a given treatment (N24h) . The results indicate that the number of bacteria of all investigated strains killed per day is equivalent when the same daily dose is administered twice a day with the MR dosage form than when given three times a day with the IR dosage form, in spite of the fact that the MR dosage form has approximately 20% lower bioavailability . Best results were obtained with the three-times a day regimen of the MR formulation . In conclusion, the present in vivo-PK/in vitro-PD simulations of the antimicrobial effects of cefaclor indicate that a twice-daily treatment with a MR formulation may offer a convenient and safe alternative to the conventional tid treatment.

Nature, 2004 May 27, 429(6990), 429 - 33
Assembly and function of a bacterial genotoxin; Nesic D et al.; The tripartite cytolethal distending toxin (CDT) induces cell cycle arrest and apoptosis in eukaryotic cells . The subunits CdtA and CdtC associate with the nuclease CdtB to form a holotoxin that translocates CdtB into the host cell, where it acts as a genotoxin by creating DNA lesions . Here we show that the crystal structure of the holotoxin from Haemophilus ducreyi reveals that CDT consists of an enzyme of the DNase-I family, bound to two ricin-like lectin domains . CdtA, CdtB and CdtC form a ternary complex with three interdependent molecular interfaces, characterized by globular, as well as extensive non-globular, interactions . The lectin subunits form a deeply grooved, highly aromatic surface that we show to be critical for toxicity . The holotoxin possesses a steric block of the CdtB active site by means of a non-globular extension of the CdtC subunit, and we identify putative DNA binding residues in CdtB that are essential for toxin activity.

J Clin Immunol, 2004 Jul, 24(4), 354 - 60
Antibody levels after regular childhood vaccinations in the immunological screening of children with recurrent otitis media; Wiertsema SP et al.; Recurrent otitis media may be related to defects in specific antibody production, as suggested previously . This might be reflected in lower antibody responses to vaccinations administered in the context of the national childhood vaccination program in children suffering from recurrent otitis media . In a cross-sectional study we determined the levels of antidiphtheria, antitetanus, anti- Haemophilus influenzae type b (anti-Hib) and antimeasles antibodies in sera of 163 children with two or more episodes of acute otitis media per year and in 143 children with repeated periods of persistent otitis media with effusion each lasting at least 3 months . The control group consisted of 521 age-matched healthy children, who were free of recurrent respiratory tract infections . Children with recurrent acute otitis media, including highly otitis-prone children, showed higher antidiphtheria and antitetanus antibody titers compared to controls . No differences were observed in anti-Hib and antimeasles antibody levels between children with recurrent acute otitis media and controls, nor did any of the antibody levels in children with persistent otitis media with effusion differ from those in controls . Therefore, the results of our study do not point toward a generalized immunological hyporesponsiveness in children with recurrent acute otitis media and persistent otitis media with effusion . Determination of antibody responses to regular vaccines is not indicative for otitis-proneness.

Vaccine, 2004 Feb 25, 22(8), 1047 - 53
Scientific challenges for the quality control and production of group C meningococcal conjugate vaccines; Jodar L et al.; Recommendations (formerly known as requirements) for meningococcal polysaccharide vaccines were adopted by the World Health Organisation (WHO) Expert Committee on Biological Standardisation in 1976 and amended in 1978 and 1981 . In clinical studies, these vaccines have been shown to have efficacy of at least 90% and have proved to be highly effective in vaccination programmes . Nevertheless, their inability to elicit protective responses in young infants or to induce good immunological memory has prevented their implementation in national infant immunisation schedules . Following the successful introduction of the Haemophilus influenzae type b conjugate (Hib) vaccines, considerable progress has been made in the development of similar conjugate vaccines based on meningococcal group C capsular polysaccharide . Controlled clinical trials have demonstrated that they induce protective levels of antibodies to group C polysaccharide in all age groups and, as T-cell dependent antigens, induce immunological memory and affinity maturation of anti-capsular antibodies . Such vaccines have been shown to offer protective immunity following the introduction of group C conjugates in the UK . The World Health Organisation has produced recommendations for the production and control of these new vaccines.

Vaccine, 2004 Feb 25, 22(8), 975 - 83
Prospective population-based incidence of Haemophilus influenzae type b meningitis in Thailand; Rerks-Ngarm S et al.; There are limited prospective data for Haemophilus influenzae type b (Hib) disease in Asia, where some countries are considering vaccine introduction . A prospective population-based study was conducted to measure the incidence of Hib meningitis in children in two northern provinces of Thailand . Children <5 years with symptoms consistent with bacterial meningitis were enrolled in the study if inclusion criteria were met . The study enrolled 598 children with clinical meningitis, 76% of whom received lumbar puncture . The rate of probable bacterial meningitis was 26.6/100,000 children <5 years per year . There were four cases of laboratory confirmed Hib meningitis (rate 3.8/100,000 children <5 years per year) . These findings suggest a relatively low incidence of Hib meningitis . However, additional data from studies of pneumonia are needed to define the Hib disease burden in Thailand.

Vet Rec, 2004 May 8, 154(19), 585 - 9
Efficacy of danofloxacin in the treatment of respiratory disease in European cattle; Rowan TG et al.; The efficacy of an injectable formulation of danofloxacin (180 mg/ml) in the treatment of naturally occurring bovine respiratory disease was evaluated in field studies on farms in France, Ireland and the United Kingdom . Cattle aged one week to 15 months with clinical respiratory disease were randomly allocated to treatment with 6 mg/kg danofloxacin or 10 mg/kg tilmicosin, administered by a single subcutaneous injection on day 0 . A second injection of danofloxacin was administered on day 2, only if predefined clinical criteria were met . Mannheimia haemolytica, Pasteurella multocida and Haemophilus somnus were isolated from pretreatment nasopharyngeal swabs taken on all the farms . After the treatment, there was a more rapid improvement in the clinical response of the 178 animals treated with danofloxacin by day 2 (P < 0.01) than in the 90 treated with tilmicosin . For both treatments, there were similar significant (P < 0.001) reductions in the mean rectal temperature and severity of clinical signs of abnormal respiration and depression, on days 4 and 10 compared with day 0; 78.1 per cent of the animals treated with danofloxacin and 78.5 per cent of those treated with tilmicosin completed the studies . Danofloxacin 18 per cent was clinically safe and as effective as tilmicosin in the treatment of bovine respiratory disease.

Arch Biochem Biophys, 2004 Jun 15, 426(2), 182 - 200
Antigenic glycans in parasitic infections: implications for vaccines and diagnostics; Nyame AK et al.; Infections by parasitic protozoans and helminths are a major world-wide health concern, but no vaccines exist to the major human parasitic diseases, such as malaria, African trypanosomiasis, amebiasis, leishmaniasis, schistosomiasis, and lymphatic filariasis . Recent studies on a number of parasites indicate that immune responses to parasites in infected animals and humans are directed to glycan determinants within cell surface and secreted glycoconjugates and that glycoconjugates are important in host-parasite interactions . Because of the tremendous success achieved recently in generating carbohydrate-protein conjugate vaccines toward microbial infections, such as Haemophilus influenzae type b, there is renewed interest in defining parasite-derived glycans in the prospect of developing conjugate vaccines and new diagnostics for parasitic infections . Parasite-derived glycans are compelling vaccine targets because they have structural features that distinguish them from mammalian glycans . There have been exciting new developments in techniques for glycan analysis and the methods for synthesizing oligosaccharides by chemical or combined chemo-enzymatic approaches that now make it feasible to generate parasite glycans to test as vaccine candidates . Here, we highlight recent progress made in elucidating the immunogenicity of glycans from some of the major human and animal parasites, the potential for developing conjugate vaccines for parasitic infections, and the possible utilization of these novel glycans in diagnostics.

Clin Infect Dis, 2004 Jun 1, 38(11), 1564 - 9 Epub 2004 May 12.
Quinolone-resistant Haemophilus influenzae in a long-term care facility: clinical and molecular epidemiology; Nazir J et al.; We describe a clonal outbreak of quinolone-resistant Haemophilus influenzae (QRHI) from an affiliated long-term care facility (LTCF-A); the outbreak was associated with the clinical use of levofloxacin, which was determined to be a risk factor for acquisition of QRHI . The minimum inhibitory concentration to which 90% of isolates were susceptible (MIC90), as determined by broth microdilution, was >4 microg/mL for levofloxacin, >2 microg/mL for moxifloxacin, >2 microg/mL for gatifloxacin, and 8 microg/mL for gemifloxacin . The MIC90, as determined by Etest (AB Biodisk), was >32 microg/mL for levofloxacin, ciprofloxacin, moxifloxacin, and gatifloxacin . Having been a resident at LTCF-A and having chronic obstructive pulmonary disease were significant risk factors for acquisition of QRHI at our 500-bed hospital (New York Hospital Queens) . All QRHI isolates were found to be genetically related by pulsed-field gel electrophoresis, were nontypeable, were susceptible to ceftriaxone and azithromycin, and were negative for beta -lactamase production . Emphasis on patient contact and respiratory isolation and placing colonized or infected patients in cohorts yielded a marked reduction in the prevalence of QRHI at LTCF-A.

Eur J Clin Microbiol Infect Dis, 2004 Jun, 23(6), 445 - 55 Epub 2004 May 20.
Rates of antimicrobial resistance among common bacterial pathogens causing respiratory, blood, urine, and skin and soft tissue infections in pediatric patients; Jones ME et al.; Antimicrobial resistance patterns among the principal bacterial pathogens from infections of the respiratory tract, blood, skin and soft tissue, and urinary tract of pediatric patients from the USA, Canada, Germany, France, and Italy were studied using the The Surveillance Network (TSN) database . Among Streptococcus pneumoniae isolates from respiratory tract infections, the prevalence of high-level penicillin resistance (MIC>/=2 microg/ml) ranged from 1.1 (Italy) to 36.2% (USA); erythromycin resistance was higher, ranging from 13.4 (Germany) to 63.8% (France) . The prevalence of beta-lactamase-positive Haemophilus influenzae among isolates from lower respiratory tract infections ranged from <10 (Italy and Germany) to 38.4% (USA) . Among isolates from blood and skin and soft tissue infections, the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) ranged from 7.2% (Canada and Germany) to 27.3% (Italy) . The prevalence of Escherichia coli and Klebsiella pneumoniae with putative extended-spectrum beta-lactamases among isolates from blood, urinary tract, and skin and soft tissue infections ranged from 0 (Germany and France) to 29.6% (Italy) . With the exception of pseudomonal infections or infections with MRSA, amoxicillin-clavulanate retained moderate activity, whilst ceftriaxone and cefepime were the most effective broad-spectrum injectable agents . Meropenem was the most effective agent against Pseudomonas aeruginosa with <5% resistance . Low levels of resistance, along with acceptable safety profiles and the availability of convenient oral formulations, continue to support the use of ceftriaxone, cefepime, amoxicillin-clavulanate, and meropenem as viable options for the treatment of infections in pediatric patients.

Infect Immun, 2004 Jun, 72(6), 3418 - 28
A novel lectin, DltA, is required for expression of a full serum resistance phenotype in Haemophilus ducreyi; Leduc I et al.; Haemophilus ducreyi, the causative agent of chancroid, is highly resistant to the complement-mediated bactericidal activity of normal human serum (NHS) . Previously, we identified DsrA (for ducreyi serum resistance A), a major factor required for expression of the serum resistance phenotype in H . ducreyi . We describe here a second outer membrane protein, DltA (for ducreyi lectin A), which also contributes to serum resistance in H . ducreyi . Isogenic dltA mutants, constructed in 35000HP wild-type and FX517 dsrA backgrounds, were more susceptible to the bactericidal effects of NHS than each respective parent, demonstrating the additive effect of the mutations . Furthermore, expression of dltA in H . influenzae strain Rd rendered this highly susceptible strain partially resistant to 5% NHS compared to a vector-control strain . Although primary basic local alignment search tool analysis of the dltA open reading frame revealed no close bacterial homologue, similarity to the beta-chain of the eukaryotic lectin ricin was noted . DltA shares highly conserved structural motifs with the ricin beta chain, such as cysteines and lectin-binding domains . To determine whether dltA was a lectin, ligand blots and affinity chromatography experiments were performed . DltA was affinity purified on immobilized lactose and N-acetylgalactosamine, and N-glycosylated but not glycosidase-treated model glycoproteins bound DltA . These data indicate that DltA is a lectin with specificity for lactose-related carbohydrates (CHO) and is important for H . ducreyi serum resistance.

Pediatr Int, 2004 Jun, 46(3), 280 - 4
Natural immunity to Haemophilus influenzae type B in children of Ankara, Turkey; Ocaktan E et al.; BACKGROUND: Haemophilus influenzae type b (Hib) infection has a high morbidity and mortality rate especially in children under 5 years of age . The incidence of Hib disease in Turkey is not known, and Hib vaccine is not included in the National Immunization Program . The aim of this study was to determine the natural immunity to Hib of children 6-60 months of age living in the Park Health Center region of Ankara, Turkey . METHODS: A total of 270 children were selected by layered random sampling method, and 242 of them (89.6%) participated in the study . A questionnaire was given to the parents of the children who were included in the study and blood samples were taken from those children . Anti-Hib IgG antibody (anti-PRP) level was determined in the serum by using anti-Haemophilus influenzae IgG EIA kit and anti-PRP antibody levels of 0.15 microg/mL and over were accepted as the natural immunity . RESULTS: Natural immunity was determined in 65.3% of the children . A relationship was determined statistically between the history of disease with possible Hib agent and with natural immunity . CONCLUSIONS: The exposure rate of children with Hib was higher than expected, even in children who were just a few months old . Our data revealed that multicentric, national studies should be done to define the burden of Hib disease before making a decision for Hib vaccine to be included in the National Immunization Program.

Lakartidningen, 2004 Apr 22, 101(17), 1488 - 92
{Microbial diagnosis with PCR will become clinically beneficial with a faster analysis}; Fohlman J et al.; PCR was introduced in 1985 by Mullis and was immediately recognized as a valuable tool in biomedical research and was awarded the Nobel Prize in 1993 . Two culture-negative meningitis cases are described where Haemophilus influenzae and Neisseria meningitidis were found by 16SRNA-PCR . The modern real time PCR technology using fluorescent probes (hybridization probes, lightup probes, molecular beacons etc) for detection of the PCR-product or on DNA microarray chips, is under development for routine use . Multiplex technology can be used to simultaneously detect multiple microorganisms as well as resistance genes . Using super-convection with ultracentrifugation high-speed PCR, results can be obtained in 10 minutes and the amplificate can also be analyzed by DNA-sequencing to achieve species identification as well as detection of resistance gene mutations . The technique has mainly been applied to viruses, but is now slowly adapted to bacteria, fungi, protozoa and helminths . PCR is especially well suited for slow growing bacteria like Mycobacteria, fastidious organisms like Bartonella and contagious agents like tularemia, but also for malaria and fungi, where the advantages in sensitivity and speed can be exploited . The limit for application to routine analysis will depend on the development of simple and fast procedures for nucleic acid extraction, as well as interpretation of the PCR analysis per se, since highly efficient thermocyclers now are on the markets.

Vaccine, 2004 Jun 2, 22(17-18), 2226 - 33
Safety, reactogenicity and immunogenicity of a combined hexavalent tetanus, diphtheria, acellular pertussis, hepatitis B, inactivated poliovirus vaccine and Haemophilus influenzae type b conjugate vaccine, for primary immunization of infants; Zepp F et al.; Safety, reactogenicity and immunogenicity of GSK Biologicals' hexavalent DTPa-HBV-IPV/Hib vaccine (Infanrix)hexa) was assessed when used for primary vaccination at 3, 4 and 5 months of age (N = 2163), compared to the separate administration of DTPa-IPV/Hib and HBV vaccines (N = 720) . A similar safety and reactogenicity profile was demonstrated for both vaccine regimens, as well as a good immune response for all antigen components . By offering protection against six diseases in a series of single injections, the hexavalent DTPa-HBV-IPV/Hib vaccine was shown to be a safe, well tolerated and immunogenic alternative to primary immunization with licensed separately administered vaccines.

Prescrire Int, 2004 Apr, 13(70), 50 - 3
Six-component vaccines: new preparations . Simpler hepatitis B vaccination of infants; Structure of serine acetyltransferase in complexes with CoA and its cysteine feedback inhibitor; Department of Biochemistry, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USASerine acetyltransferase (SAT, EC 2.3.1.30) catalyzes the CoA-dependent acetylation of the side chain hydroxyl group of l-serine to form O-acetylserine, as the first step of a two-step biosynthetic pathway in bacteria and plants leading to the formation of l-cysteine . This reaction represents a key metabolic point of regulation for the cysteine biosynthetic pathway due to its feedback inhibition by cysteine . We have determined the X-ray crystal structure of Haemophilus influenzae SAT in complexes with CoA and its cysteine feedback inhibitor . The enzyme is a 175 kDa hexamer displaying the characteristic left-handed parallel beta-helix (LbetaH) structural domain of the hexapeptide acyltransferase superfamily of enzymes . Cysteine is bound in a crevice between adjacent LbetaH domains and underneath a loop excluded from the coiled LbetaH . The proximity of its thiol group to the thiol group of CoA derived from superimposed models of the cysteine and CoA complexes confirms that cysteine is bound at the active site . Analysis of the contacts of SAT with cysteine and CoA and the conformational differences that distinguish these complexes provides a structural basis for cysteine feedback inhibition, which invokes competition between cysteine and serine binding and a cysteine-induced conformational change of the C-terminal segment of the enzyme that excludes binding of the cofactor.

Ned Tijdschr Geneeskd, 2004 Apr 24, 148(17), 836 - 40
{Community-acquired pneumonia: pathogens and course in patients admitted to a general hospital}; Braun JJ et al.; OBJECTIVE: To determine prognostic factors and the significance of (non-invasive) microbiological tests for the clinical course of patients admitted to one general hospital with community-acquired pneumonia (CAP) . DESIGN: Prospective observational study . METHOD: Patients admitted to one location of a general hospital with symptoms of CAP during the period January 1998-December 1999 were included . Data from the anamnesis, physical examination and laboratory tests were recorded and samples were taken for diagnosis of the possible pathogens . Cultures were made of sputum and blood, serum and sputum were examined for infection with viral and atypical microorganisms, and the urine was screened for Streptococcus pneumoniae and Legionella antigens . RESULTS: Of the 157 patients studied, 28 (18%) died as a direct result of CAP . In a stepwise multivariate analysis, age > or = 65 years, increased serum-creatinine levels and hypercapnia were independent predictors of mortality . Streptococcus pneumoniae and Haemophilus influenzae were detected 53 and 19 times, respectively, and were the bacterial pathogens most frequently found . Among the nonbacterial pathogens, Mycoplasma pneumoniae was found 37 times and serologic tests for influenza A or B virus were positive in 34 cases . In 62 patients (39%), extensive microbiological tests revealed signs of a combined infection and in 20 patients (13%) no microorganism could be detected . An elevated serum-procalcitonin level was associated with bacterial infections . As a consequence of their retrospective nature, the results of the extensive serological tests did not contribute to reaching a diagnosis in daily clinical practice . In comparison with the literature, there was a markedly high number of subclinical or atypical infections with Legionella (8%) and Bordetella (18%) and a high incidence of viral and atypical microorganisms as the cause of CAP.

Diagn Microbiol Infect Dis, 2004 May, 49(1), 31 - 6
In vitro selection of resistance in haemophilus influenzae by 4 quinolones and 5 beta-lactams; Clark C et al.; We tested abilities of ciprofloxacin, levofloxacin, gatifloxacin, moxifloxacin, amoxicillin, amoxicillin/clavulanate, cefixime, cefpodoxime, and cefdinir to select resistant mutants in 5 beta-lactamase positive and 5 beta-lactamase negative Haemophilus influenzae strains by single and multistep methodology . In multistep tests, amoxicillin, amoxicillin/clavulanate and cefpodoxime exposure did not cause >4-fold minimum inhibitory concentration (MIC) increase after 50 days . One mutant selected by cefdinir had one amino acid substitution (Gly490Glu) in PBP3 and became resistant to cefdinir . Cefixime exposure caused 8-fold MIC-increase in 1 strain with TEM but the mutant remained cefixime susceptible and had no alteration in PBP3 or TEM . Among 10 strains tested, ciprofloxacin, moxifloxacin, gatifloxacin, levofloxacin caused >4-fold MIC increase in 6, 6, 5, and 2 strain, respectively . Despite the increases in quinolone MICs, none of the mutants became resistant to quinolones by established criteria . Quinolone selected mutants had quindone resistance-determining region (QRDR) alterations in GyrA, GyrB, ParC, ParE . Four quinolone mutants had no QRDR alterations . Among beta-lactams cefdinir and cefixime selected one mutant each with higher MICs however amoxicillin, amoxicillin/clavulanate, and cefpodoxime exposure did not select resistant mutants.

J Microbiol Methods, 2004 Jun, 57(3), 421 - 4
Construction of a nontypeable Haemophilus influenzae-specific ectopic delivery vector; Daines DA et al.; Complementation of chromosomal mutations in trans can introduce artifacts due to the number of episomal copies of the gene in question . One solution is to study the gene expressed at a single ectopic site in cis . We have designed and constructed a vector that allows homologous recombination into a gene encoding a frame-shifted IS1016-V6 protein in the Haemophilus influenzae Rd KW20 chromosome (HI1018) . This site is the location of the > or = 35 kilobase capsule locus in encapsulated type b and d strains . This locus is not present in the nontypeable Rd KW20 strain, thus allowing ectopic expression of genes homologously recombined into HI1018 without polar effects.

Pediatr Infect Dis J, 2004 May, 23(5), 406 - 13
Relationship among peripheral leukocyte counts, etiologic agents and clinical manifestations in acute otitis media; Polachek A et al.; OBJECTIVES: To analyze the peripheral leukocyte counts of children with acute otitis media (AOM) in relation to etiology, age, clinical symptoms and signs, prior antibiotic treatment, previous AOM history and pathogen eradication during antibiotic therapy . PATIENTS AND METHODS: Leukocyte counts were determined at diagnosis and on days 4 to 6 of therapy in patients age 3 to 36 months with AOM enrolled in double tympanocentesis studies . Clinical status was determined by a clinical score evaluating severity of fever, irritability and tympanic membrane redness and bulging . RESULTS: Of 771 enrolled patients, culture-positive middle ear fluid was reported in 590 (77%): 294 (50%) Haemophilus influenzae; 150 (25%) Streptococcus pneumoniae; 127 (21%) H . influenzae and S . pneumoniae together; 9 (2%) Moraxella catarrhalis; and 10 (2%) others . Mean leukocyte count +/- sd in patients with AOM caused by S . pneumoniae (15.7 +/- 6.7 cells x 10/mm) was significantly higher than those of patients with AOM caused by H . influenzae (13.7 +/- 5.8 cells x 10/mm) and patients with culture-negative AOM (13.8 +/- 5.6 cells x 10/mm), P < 0.01 for each comparison . Mean absolute neutrophil count (ANC) +/- sd was higher in patients with AOM caused by S . pneumoniae (8.6 +/- 5.1 cells x 10/mm) than in patients with AOM caused by H . influenzae (6.5 +/- 4.0 cells x 10/mm) or culture-negative patients (6.5 +/- 4.4 cells x 10/mm), P < 0.05 for each comparison . When multivariant regression analysis was used, S . pneumoniae etiology and fever (body temperature > or = 38 degrees Celsius) were independent factors, each significantly associated with leukocyte and ANCs . A significant correlation was found between leukocyte and ANCs and higher clinical scores in patients with pneumococcal AOM (P = 0.01, r = 0.21 and P < 0.01, r = 0.27, respectively) . The mean leukocyte count on Days 4 to 6 was lower than on Day 1 (11.5 +/- 4.1 compared with 14.2 +/- 6.0 cells x 10/mm, P < 0.01) . When paired (Day 1 and Days 4 to 6) examinations were analyzed, the leukocyte counts were lower on Days 4 to 6 irrespective of bacterial eradication or persistence . CONCLUSIONS: Significantly higher leukocyte counts and ANCs were found in pneumococcal AOM than in AOM caused by H . influenzae or in culture-negative AOM . A significant decrease in leukocyte counts was found during antibiotic therapy for AOM, regardless of etiology and bacteriologic outcome . Isolation of S . pneumoniae and fever were each significantly associated with increased peripheral leukocyte and ANCs.

Zhonghua Jie He He Hu Xi Za Zhi, 2004 Mar, 27(3), 155 - 60
{Resistance surveillance of common community respiratory pathogens isolated in China, 2002 - 2003}; Wang H et al.; OBJECTIVE: To investigate antimicrobial resistance of common community respiratory pathogens isolated in China, 2002 - 2003 . METHODS: 779 strains of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Group A beta-haemolytic Streptococci and oxacillin-susceptible Staphylococcus aureus (MSSA) were isolated from patients with community-acquired respiratory tract infections at 5 hospitals in China from April 2002 to 2003 . Meanwhile, 185 strains of S . pneumoniae, H . influenzae and M . catarrhalis were isolated from nasopharynx swabs at 2 day-care centers in Beijing . Agar dilution method was used to determine the minimal inhibitory concentration (MICs) of cefprozil and other 9 antibiotics against these strains . RESULTS: The prevalence of penicillin-intermediate S . pneumoniae (PISP) was 23.9% and that of penicillin-resistant S . pneumoniae (PRSP) was 22.7% at 5 cities in China . The prevalence of PISP were 44.1% in Hangzhou, 26.2% in Wuhan, 21.5% in Shenyang, 20.8% in Shanghai, 18.5% in Beijing, and 12.7% at day-care centers in Beijing;the prevalance of PRSP were 34.9% in day-care centers, 31.9% in Shanghai, 27.9% in Wuhan, 22.1% in Hangzhou, 13.8% in Shenyang and 8.6% in Beijing . The susceptible rate of levofloxacin in S . pneumoniae was 96.3% . 9.5% of H . influenzae and 87.4% of M . catarrhalis produced beta-lactamases . The susceptibility of amoxicillin/clavulanate, cefaclor, cefprozil, cefuroxime, ceftriaxone, azithromycin, and levofloxain in these two species ranged from 96.4% to 100% . The resistance rate of azithromycin in S . pneumoniae was higher than 60% . Cefprozil MICs against PISP, Group A beta-haemolytic Streptococci and MSSA were 4 - 16 fold lower than cefaclor . CONCLUSIONS: Antimicrobial resistance in respiratory pathogens, especially S . pneumoniae is increasing . It brings concerns that high macrolide resistance was found in gram-positive cocci . Cefprozil was more active than cefaclor against respiratory pathogens.

Neth J Med, 2004 Feb, 62(2), 45 - 52
Prevention of infections in hyposplenic and asplenic patients: an update; Melles DC et al.; Patients with functional or anatomic asplenia are at a significantly increased risk of overwhelming infection, particularly involving the encapsulated bacteria Streptococcus pneumoniae and Haemophilus influenzae . The risk is highest in infants and young children, but adults also have an increased risk of infection . Preventive strategies are very important and fall into three major categories: immunoprophylaxis, antibiotic prophylaxis and education . Studies have shown that many asplenic patients are unaware of their increased risk for serious infection and the appropriate health precautions that should be undertaken . In this article we emphasise the need for preventive measures in hyposplenic and asplenic patients . We discuss the value of newly developed conjugate vaccines and the need for revaccination . Finally we draw up a recommendation for the preventive management in functional and anatomical asplenic patients.

J Bacteriol, 2004 May, 186(10), 2928 - 35
Mutations in Haemophilus influenzae mismatch repair genes increase mutation rates of dinucleotide repeat tracts but not dinucleotide repeat-driven pilin phase variation rates; Bayliss CD et al.; High-frequency, reversible switches in expression of surface antigens, referred to as phase variation (PV), are characteristic of Haemophilus influenzae . PV enables this bacterial species, an obligate commensal and pathogen of the human upper respiratory tract, to adapt to changes in the host environment . Phase-variable hemagglutinating pili are expressed by many H . influenzae isolates . PV involves alterations in the number of 5' TA repeats located between the -10 and -35 promoter elements of the overlapping, divergently orientated promoters of hifA and hifBCDE, whose products mediate biosynthesis and assembly of pili . Dinucleotide repeat tracts are destabilized by mismatch repair (MMR) mutations in Escherichia coli . The influence of mutations in MMR genes of H . influenzae strain Rd on dinucleotide repeat-mediated PV rates was investigated by using reporter constructs containing 20 5' AT repeats . Mutations in mutS, mutL, and mutH elevated rates approximately 30-fold, while rates in dam and uvrD mutants were increased 14- and 3-fold, respectively . PV rates of constructs containing 10 to 12 5' AT repeats were significantly elevated in mutS mutants of H . influenzae strains Rd and Eagan . An intact hif locus was found in 14 and 12% of representative nontypeable H . influenzae isolates associated with either otitis media or carriage, respectively . Nine or more tandem 5' TA repeats were present in the promoter region . Surprisingly, inactivation of mutS in two serotype b H . influenzae strains did not alter pilin PV rates . Thus, although functionally analogous to the E . coli MMR pathway and active on dinucleotide repeat tracts, defects in H . influenzae MMR do not affect 5' TA-mediated pilin PV.

BMC Infect Dis . 2004 May 05;4(1):12.
Antimicrobial activity of innate immune molecules against Streptococcus pneumoniae, Moraxella catarrhalis and nontypeable Haemophilus influenzae; Lee HY et al.; BACKGROUND: Despite its direct connection to the nasopharynx which harbors otitis media pathogens as part of its normal flora, the middle ear cavity is kept free of these bacteria by as yet unknown mechanisms . Respiratory mucosal epithelia, including those of the middle ear and eustachian tube, secrete antimicrobial effectors including lysozyme, lactoferrin and beta defensins-1 and -2 . To elucidate the role of these innate immune molecules in the normal defense and maintenance of sterility of respiratory mucosa such as that of the middle ear, we assessed their effect on the respiratory pathogens nontypeable Haemophilus influenzae (NTHi) 12, Moraxella catarrhalis 035E, and Streptococcus pneumoniae 3, and 6B . METHODS: Two assay methods, the radial assay and the liquid broth assay, were employed for testing the antimicrobial activity of the molecules . This was done in order to minimize the possibility that the observed effects were artifacts of any single assay system employed . Also, transmission electron microscopy (TEM) was employed to evaluate the effect of antimicrobial innate immune molecules on OM pathogens . For the statistical analysis of the data, Student's t-test was performed . RESULTS: Results of the radial diffusion assay showed that beta defensin-2 was active against all four OM pathogens tested, while treatment with beta defensin-1 appeared to only affect M . catarrhalis . The radial assay results also showed that lysozyme was quite effective against S . pneumoniae 3 and 6B and was partially bacteriostatic/bactericidal against M . catarrhalis . Lysozyme however, appeared not to affect the growth of NTHi . Thus, lysozyme seems to have a more pronounced impact on the growth of the Gram-positive S . pneumoniae as compared to that of Gram-negative pathogens . Lactoferrin on the other hand, enhanced the growth of the bacteria tested . The results of the radial assays were confirmed using liquid broth assays for antimicrobial activity, and showed that lysozyme and beta defensin-2 could act synergistically against S . pneumoniae 6B . Moreover, in the liquid broth assay, beta defensin-1 showed a modest inhibitory effect on the growth of S . pneumoniae 6B . As assessed by ultrastructural analysis, lysozyme and beta defensin-2, and to a much lesser extent, beta defensin-1, appeared to be able to cause damage to the bacterial membranes . CONCLUSIONS: Here we report that lysozyme and the beta defensins can inhibit the growth of clinical isolates of otitis media pathogens - namely NTHi strain 12, S . pneumoniae strains 3 and 6B and M . catarrhalis strain 035E - and cause ultrastructural damage to these pathogens . Moreover, we demonstrate that lysozyme and beta defensin-2 can act synergistically against S . pneumoniae . These findings are consistent with the concept that secreted antimicrobial peptides and other components of innate immunity constitute the first line of defense protecting host mucosal surfaces, including the tubotympanal (eustachian tube and middle ear cavity) mucosa, against pathogens.

Nucleic Acids Res, 2004 Apr 30, 32(8), 2353 - 61 Print 2004.
Identification and functional analysis of 'hypothetical' genes expressed in Haemophilus influenzae; Kolker E et al.; The progress in genome sequencing has led to a rapid accumulation in GenBank submissions of uncharacterized 'hypothetical' genes . These genes, which have not been experimentally characterized and whose functions cannot be deduced from simple sequence comparisons alone, now comprise a significant fraction of the public databases . Expression analyses of Haemophilus influenzae cells using a combination of transcriptomic and proteomic approaches resulted in confident identification of 54 'hypothetical' genes that were expressed in cells under normal growth conditions . In an attempt to understand the functions of these proteins, we used a variety of publicly available analysis tools . Close homologs in other species were detected for each of the 54 'hypothetical' genes . For 16 of them, exact functional assignments could be found in one or more public databases . Additionally, we were able to suggest general functional characterization for 27 more genes (comprising approximately 80% total) . Findings from this analysis include the identification of a pyruvate-formate lyase-like operon, likely to be expressed not only in H.influenzae but also in several other bacteria . Further, we also observed three genes that are likely to participate in the transport and/or metabolism of sialic acid, an important component of the H.influenzae lipo-oligosaccharide . Accurate functional annotation of uncharacterized genes calls for an integrative approach, combining expression studies with extensive computational analysis and curation, followed by eventual experimental verification of the computational predictions.

Am J Physiol Lung Cell Mol Physiol, 2004 Aug, 287(2), L428 - 37 Epub 2004 Apr 30.
Endotoxin responsiveness of human airway epithelia is limited by low expression of MD-2; Jia HP et al.; The expression of inducible antimicrobial peptides, such as human beta-defensin-2 (HBD-2) by epithelia, comprises a component of innate pulmonary defenses . We hypothesized that HBD-2 induction in airway epithelia is linked to pattern recognition receptors such as the Toll-like receptors (TLRs) . We found that primary cultures of well-differentiated human airway epithelia express the mRNA for TLR-4, but little or no MD-2 mRNA, and display little HBD-2 expression in response to treatment with purified endotoxin +/- LPS binding protein (LBP) and soluble CD14 . Expression of endogenous MD-2 by transduction of airway epithelial cells with an adenoviral vector encoding MD-2 or extracellular addition of recombinant MD-2 both increased the responses of airway epithelia to endotoxin + LBP and sCD14 by >100-fold, as measured by NF-kappaB-luciferase activity and HBD-2 mRNA expression . MD-2 mRNA could be induced in airway epithelia by exposure of these cells to specific bacterial or host products (e.g., killed Haemophilus influenzae, the P6 outer membrane protein from H . influenzae, or TNF-alpha + IFN-gamma) . These findings suggest that MD-2, either coexpressed with TLR-4 or secreted when produced in excess of TLR-4 from neighboring cells, is required for airway epithelia to respond sensitively to endotoxin . The regulation of MD-2 expression in airway epithelia and pulmonary macrophages may serve as a means to modify endotoxin responsiveness in the airway.

Vaccine, 2004 May 7, 22(15-16), 2018 - 22
Catch-up vaccination against Haemophilus influenzae type b in human immunodeficiency virus-infected Thai children older than 2 years old; Chokephaibulkit K et al.; Although most of Thai children older than 2 years are immune against Haemophilus influenzae type b (Hib) without prior vaccination, it may not be the case in HIV-infected children . Of 44 HIV-infected children tested before vaccination at the mean age of 36 months (range 24-84 months), 32 (73%) were susceptible (anti-PRP <0.15 microg/ml) . At 6 months after a single dose of tetanus-conjugated Hib vaccination, 67% developed anti-PRP >/=0.15 microg/ml, however, only 33% developed titer of >/=1 microg/ml . Four of seven (57%) with anti-PRP 0.15-0.99 microg/ml at baseline were boosted to the titer of >/=1 microg/ml after vaccination . Seroconversion rate and geometric mean titer (GMT) level in response to the vaccination did not correlate with HIV stage, but did correlate with viral load level of 100,000 copies/ml . HIV-infected children older than 2 years would benefit from Hib vaccination, although, one dose catch-up schedule is not sufficient in a third of these children . A second dose is needed in these children especially those with viral load of level of >100,000 copies/ml.

Jpn J Infect Dis, 2004 Apr, 57(2), 58 - 9
Endotoxin content in Haemophilus influenzae type b vaccine; Ochiai M et al.; Haemophilus influenzae type b (Hib) is a major cause of bacterial meningitis among children . Hib conjugate vaccines have effectively prevented Hib infection, and routine immunization with Hib conjugate vaccine has diminished the incidence of the disease in the United States and European countries . Introduction of Hib conjugate vaccines is also required in Japan . However, endotoxin that can carry over from Gram-negative H . influenzae with a purified component may contribute to adverse events following Hib vaccination . In the present study, we examined the endotoxin content in Hib conjugate vaccines . The Hib conjugate vaccine batches, which were produced by a European vaccine manufacturer, were shown to have considerably high endotoxin activity and to vary from 13.9 to 173.7 endotoxin units/dose . These results suggest that it is necessary to monitor the endotoxin content of the vaccine batches to ensure the quality and safety of the vaccines.

Am J Respir Crit Care Med, 2004 Aug 1, 170(3), 266 - 72 Epub 2004 Apr 29.
Persistent colonization by Haemophilus influenzae in chronic obstructive pulmonary disease; Murphy TF et al.; Nontypeable Haemophilus influenzae colonizes the respiratory tract of adults with chronic obstructive pulmonary disease (COPD) and causes intermittent exacerbations . Isolates of H . influenzae collected monthly in a prospective study were subjected to molecular typing . During a 7-year study spanning 345 patient-months of observation, 122 episodes of negative cultures lasting 1 month or more, and that were preceded and followed by isolation of an apparently identical strain of H . influenzae, were found . Seventeen such episodes of negative cultures, lasting 6 months or more and spanning 203 patient-months, were studied in detail to test the hypothesis that these periods of negative cultures represented continuous colonization by the same strain of H . influenzae . Molecular typing by three independent methods established that the strains preceding and following the episodes of negative cultures were indeed identical . Strain-specific H . influenzae DNA was detected in some of the sputum samples that had yielded negative cultures . These results indicate that some patients with COPD are persistently colonized with H . influenzae and that sputum cultures underestimate the frequency of colonization of the respiratory tract by H . influenzae in COPD . This observation has a significant impact on understanding bacterial colonization in COPD.

Jpn J Antibiot, 2004 Feb, 57(1), 105 - 17
{Isolation-frequency and antibiotic susceptibility of Streptococcus pneumoniae, Haemophilus influenzae, and Pseudomonas aeruginosa isolated in regional core hospitals in the Nagaoka district of Niigata Prefecture (changes for 3 years)}; Iwashima A et al.; We measured MICs of various antimicrobial agents against Streptococcus pneumoniae, Haemophilus influenzae, and Pseudomonas aeruginosa isolated in the Nagaoka district of Niigata Prefecture in 2000 (March-May), 2001 (January-May), and 2002 (March-May) . S . pneumoniae: Fifty-six strains were isolated in 2000, 119 strains in 2001, and 88 strains in 2002 . In 2000, 2001, and 2002, 24 strains (42.9%), 58 strains (48.7%), and 40 strains (45.5%), respectively, were penicillin-intermediate S . pneumoniae (PISP), and 4 strains (7.1%), 12 strains (10.1%), and 7 strains (8.0%), respectively, were penicillin-resistant S . pneumoniae (PRSP) . Carbapenems had the most excellent antimicrobial activity, followed by penicillin G, against penicillin-susceptible S . pneumoniae (PSSP), PISP, and PRSP . H . influenzae: Seventy-six strains were isolated in 2000, 154 strains in 2001, and 91 strains in 2002 . In 2000, 2001, and 2002, 6 strains (7.9%), 8 strains (5.2%), and 7 strains (7.7%), respectively, were beta-lactamase-producing ampicillin (ABPC)-resistant strains (MIC > or = 2 micrograms/ml), showing no increase, and 14 strains (18.4%), 70 strains (45.5%), and 31 strains (34.1%), respectively, were beta-lactamase-non-producing ABPC-resistant strains (MIC > or = 2 micrograms/ml), showing a slight increase . Ceftriaxon, meropenem (MEPM), and levofloxacin had excellent antimicrobial activity against these resistant strains . P . aeruginosa: In 2000, 2001, and 2002, 135, 74, and 91 strains, respectively, were isolated, and 14 strains (10.4%), 17 strains (23.0%), and 24 strains (26.4%), respectively, were imipenem-resistant (MIC > or = 16 micrograms/ml), showing a slight increase . MEPM, biapenem, and ciprofloxacin had excellent antimicrobial activity against P . aeruginosa.

Lancet, 2004 Apr 24, 363(9418), 1386 - 8
Malaria intermittent preventive treatment in infants, chemoprophylaxis, and childhood vaccinations; Rosen JB et al.; CONTEXT: Malaria accounts for 1-3 million deaths yearly worldwide, mostly in children under 5 years of age in sub-Saharan Africa . Laboratory and clinical studies show an association between acute malaria and a decreased response to diphtheria and tetanus toxoids and to meningococcal, salmonella, and Haemophilus influenzae type b vaccinations . Malaria treatment, chemoprophylaxis, or other forms of parasite suppression might improve the immune response to childhood vaccinations . However, the antimalarial 4-aminoquinolones are immunodepressive, such that antimalarial drugs might depress the vaccine response . STARTING POINT: Last year, Julius Massaga and colleagues reported a randomised double-blinded placebo-controlled study in 291 infants aged 12-16 weeks in Tanzania (Lancet 2003; 361: 1853-60) . At enrollment, children received their third dose of combined diphtheria-tetanus-pertussis and poliomyelitis expanded vaccines with the first of three daily doses of amodiaquine intermittent preventive treatment (IPTi) or placebo . After 60 days, children receiving amodiaquine had significantly fewer malaria fevers than controls . WHERE NEXT: The increasing concordance of malaria control and vaccination, movement toward co-administration of IPTi with immunisation, and the increase in travellers to malarious areas who receive concurrent vaccinations and chemoprophylaxis warrant further study.

Tohoku J Exp Med, 2004 Apr, 202(4), 245 - 54
Serotypes of Haemophilus influenzae strains isolated from pediatric patients with respiratory tract infections; Luong DC et al.; Capsular serotypes of 122 respiratory Haemophilus influenzae strains were identified utilizing antigenic examinations (Slide Agglutination Test {SAT} and counter immuno-electrophoresis {CIE}), and PCR method . Results of the two methods were compared to find disagreement . Clinical and bacteriological characteristics of encapsulated strains were analyzed . By utilizing PCR assay, 3 (2.5%) serotype b, 1 (0.8%) serotype f and 118 (96.7%) non-typeable strains were found among 122 tested strains . Discrepancy between the results of antigenic examinations and PCR assay was found in one strain . That strain gave positive results by antigenic tests with capsular type c antiserum but it was determined to be capsular type f by PCR . No serotype b- strain was detected . All the encapsulated strains displayed biotype I . All the serotype b strains produced beta-lactamase, but none of those was sulbactam/ampicillin and amoxicillin/clavulanic acid resistant . The serotype f strain was beta-lactamase non-producing ampicillin susceptible . The patients suffered from respiratory infections due to encapsulated strains displayed safe clinical manifestations and good clinical responses to antibiotic treatments using beta-lactams . In pre-vaccination era, PCR is considered to be a useful method for determination of serotypes of respiratory H . influenzae, which may contribute to reducing a possibility of clonal transmission of serotype b strains among children community, which is one of the potential risk factors for pediatric invasive infections.

Aust N Z J Public Health, 2004 Feb, 28(1), 68 - 71
Getting it right--the Australian Childhood Immunisation Register and immunisation rates in south-eastern Sydney; Botham SJ et al.; OBJECTIVES: To assess the completeness of ACIR data for south-eastern Sydney children at 12 to < 15 months of age and to develop strategies to improve rates . METHODS: We surveyed children aged 12 to < 15 months listed as overdue on the ACIR011A report, December 2001, by contacting the last immunisation provider or the parents . RESULTS: From the 470 children listed as overdue, 162 children were systematically selected to form the study group . Seventeen were lost to follow-up and results are available for 145 children . Eighty children were up to date (69, encounter forms incorrect or not sent; 11, vaccinated overseas) and 11 children had moved overseas . Fifty-four children were overdue (30, provider error due to incorrect Haemophilus influenzae type b (Hib) schedule or Hib vaccine omitted; 18 did not complete schedule; 6 conscientious objectors) . CONCLUSIONS: Our study indicates that between 6-9% of children on the register were likely to be overdue, whereas the ACIR011A report identified 19% . Failure of immunisation providers to correctly complete, or send encounter forms to the ACIR was the main reason for this discrepancy . Migration and failure to record overseas vaccination were also factors . Implications: Public health units should develop good working relationships with immunisation providers to assist and encourage immunisation and the completion and submission of encounter forms . A quarterly review, using the third dose assumption, of all children aged 12 to < 15 months identified on the ACIR011A report as overdue for immunisation may be an effective way for public health units to increase apparent rates for their area.

OMICS, 2004 Spring, 8(1), 25 - 41
In Silico Metabolic Model and Protein Expression of Haemophilus influenzae Strain Rd KW20 in Rich Medium; Raghunathan A et al.; The intermediary metabolism of Haemophilus influenzae strain Rd KW20 was studied by a combination of protein expression analysis using a recently developed direct proteomics approach, mutational analysis, and mathematical modeling . Special emphasis was placed on carbon utilization, sugar fermentation, TCA cycle, and electron transport of H . influenzae cells grown microaerobically and anaerobically in a rich medium . The data indicate that several H . influenzae metabolic proteins similar to Escherichia coli proteins, known to be regulated by low concentrations of oxygen, were well expressed in both growth conditions in H . influenzae . An in silico model of the H . influenzae metabolic network was used to study the effects of selective deletion of certain enzymatic steps . This allowed us to define proteins predicted to be essential or non-essential for cell growth and to address numerous unresolved questions about intermediary metabolism of H . influenzae . Comparison of data from in vivo protein expression with the protein list associated with a genome-scale metabolic model showed significant coverage of the known metabolic proteome . This study demonstrates the significance of an integrated approach to the characterization of H . influenzae metabolism.

Antimicrob Agents Chemother, 2004 May, 48(5), 1630 - 9
Genetic and molecular characterization of beta-lactamase-negative ampicillin-resistant Haemophilus influenzae with unusually high resistance to ampicillin; Kaczmarek FS et al.; Previous studies with beta-lactamase-negative, ampicillin-resistant (BLNAR) Haemophilus influenzae from Japan, France, and North America indicate that mutations in ftsI encoding PBP3 confer ampicillin MICs of 1 to 4 micro g/ml . Several BLNAR strains with ampicillin MICs of 4 to 16 micro g/ml recently isolated from North America were studied . Pulsed-field gel electrophoresis identified 12 unique BLNAR strains; sequencing of their ftsI transpeptidase domains identified 1 group I and 11 group II mutants, as designated previously (K . Ubukata, Y . Shibasaki, K . Yamamoto, N . Chiba, K . Hasegawa, Y . Takeuchi, K . Sunakawa, M . Inoue, and M . Konno, Antimicrob . Agents Chemother . 45:1693-1699, 2001) . Geometric mean ampicillin MICs for several clinical isolates were 8 to 10.56 micro g/ml . Replacement of the ftsI gene in H . influenzae Rd with the intact ftsI from several clinical isolates resulted in integrants with typical BLNAR geometric mean ampicillin MICs of 1.7 to 2.2 micro g/ml . Cloning and purification of His-tagged PBP3 from three clinical BLNAR strains showed significantly reduced Bocillin binding compared to that of PBP3 from strain Rd . Based on these data, changes in PBP3 alone could not account for the high ampicillin MICs observed for these BLNAR isolates . In an effort to determine the presence of additional mechanism(s) of ampicillin resistance, sequencing of the transpeptidase regions of pbp1a, -1b, and -2 was performed . While numerous changes were observed compared to the sequences from Rd, no consistent pattern correlating with high-level ampicillin resistance was apparent . Additional analysis of the resistant BLNAR strains revealed frame shift insertions in acrR for all four high-level, ampicillin-resistant isolates . acrR was intact for all eight low-level ampicillin-resistant and four ampicillin-susceptible strains tested . A knockout of acrB made in one clinical isolate (initial mean ampicillin MIC of 10.3 micro g/ml) lowered the ampicillin MIC to 3.67 micro g/ml, typical for BLNAR strains . These studies illustrate that BLNAR strains with high ampicillin MICs exist that have combined resistance mechanisms in PBP3 and in the AcrAB efflux pump.

Antimicrob Agents Chemother, 2004 May, 48(5), 1509 - 14
Rapidly increasing prevalence of beta-lactamase-nonproducing, ampicillin-resistant Haemophilus influenzae type b in patients with meningitis; Hasegawa K et al.; A total of 395 Haemophilus influenzae strains from 226 Japanese institutions participating in the Nationwide Surveillance Study Group for Bacterial Meningitis were received from 1999 to 2002 . All strains were analyzed by PCR to identify the resistance genes, and their susceptibilities to beta-lactam agents were determined . Of these strains, 29.1% were beta-lactamase nonproducing and ampicillin (AMP) susceptible (BLNAS) and lacked all resistance genes; 15.4% were beta-lactamase producing and AMP resistant and had the bla(TEM-1) gene; 30.6% were beta-lactamase nonproducing and AMP resistant (low-BLNAR) and had a Lys-526 or His-517 amino acid substitution in ftsI encoding PBP 3; 13.9% were beta-lactamase nonproducing and AMP resistant (BLNAR) and had an additional substitution of Thr-385 in ftsI; 9.1% were amoxicillin-clavulanic acid resistant (BLPACR I) and had the bla(TEM-1) gene and a Lys-526 or His-517 amino acid substitution in ftsI; and 1.8% showed resistance similar to that of the BLPACR I group (BLPACR II) but had bla(TEM-1) gene and ftsI substitutions, as was the case for the BLNAR strains . All but three strains were serotype b . The prevalence of BLNAR strains has increased rapidly: 0% in 1999, 5.8% in 2000, 14.1% in 2001, and 21.3% in 2002 . The MICs at which 90% of BLNAR isolates were inhibited were as follows: AMP, 16 micro g/ml; cefotaxime, 1 micro g/ml; ceftriaxone, 0.25 micro g/ml; and meropenem, 0.5 micro g/ml . All of these values were higher than those for the BLNAS counterpart strains . The relatively wide distributions of the beta-lactam MICs for BLNAR strains presumably reflect variations in ftsI gene mutations . Pulsed-field gel electrophoresis suggested the rapid spread of specific H . influenzae type b strains throughout Japan . Expedited vaccination, rapid identification, and judicious antibiotic use could slow their spread.

Cell Microbiol, 2004 Jun, 6(6), 555 - 67
Different meningitis-causing bacteria induce distinct inflammatory responses on interaction with cells of the human meninges; Fowler MI et al.; The interactions of bacterial pathogens with cells of the human leptomeninges are critical events in the progression of meningitis . An in vitro model based on the culture of human meningioma cells was used to investigate the interactions of the meningeal pathogens Escherichia coli K1, Haemophilus influenzae, Neisseria meningitidis and Streptococcus pneumoniae . A rank order of association with meningioma cells was observed, with N . meningitidis showing the highest levels of adherence, followed by E . coli, S . pneumoniae and H . influenzae . Neisseria meningitidis and H . influenzae did not invade meningioma cells or induce cell death, but induced a concentration-dependent secretion of inflammatory mediators . Neisseria meningitidis induced higher levels of IL-6, MCP-1, RANTES and GM-CSF than H . influenzae, but there was no significant difference in the levels of IL-8 induced by both pathogens . Streptococcus pneumoniae was also unable to invade meningioma cells, but low concentrations of bacteria failed to stimulate cytokine secretion . However, higher concentrations of pneumococci led to cell death . By contrast, only E . coli K1 invaded meningioma cells directly and induced rapid cell death before an inflammatory response could be induced . These data demonstrate that the interactions of different bacterial pathogens with human meningeal cells are distinct, and suggest that different intervention strategies may be needed in order to prevent the morbidity and mortality associated with bacterial meningitis.

Med Dosw Mikrobiol, 2003, 55(4), 357 - 63
{Application of PCR to Haemophilus influenzae typing . I . PCR standardization for bexA gene fragment detection and for detection of DNA fragment specific for H.influenzae type B}; Zasada AA et al.; PCR standardization was performed in order to detect a fragment of bexA gene, which is presented in all capsulate H . influenzae isolates, and a DNA fragment specific for H . influenzae type b . Implementing of such PCR into H . influenzae typing may be very useful in cases of isolates from clinical material for which serotyping alone gives unclear results . Standardization of PCR detecting DNA fragments specific for all capsular types will enable to perform complete typing of H . influenzae isolates.

Kansenshogaku Zasshi, 2004 Mar, 78(3), 253 - 61
{Clinical study of 28 children with bacterial meningitis}; Ogasawara N et al.; A retrospective study was conducted on 28 patients with bacterial meningitis who were admitted to our department between April 1988 and March 2002 . The most commonly detected pathogen was group B Streptococcus in those under 1 month of age and Haemophilus influenzae (72.2%) among those over 1 month . The most commonly administered antibiotic combination (67.9%) at the initial treatment was that of cefotaxime (CTX) and ampicillin (ABPC) . We encountered one case that was resistant to both CTX and ABPC . Through this experience, it became apparent that for the initial treatment of bacterial meningitis in infants, it is necessary to apply a combination of two antibiotics, instead of a single agent, and new antibiotics should be considered for such combinations rather than persisting on conventional CTX and ABPC . The aforementioned 28 patients were divided into 2 groups--7 patients (25.0%) with sequelae and 21 (75.0%) without--and various factors noted during the diagnosis were evaluated retrospectively . It was found that the number of days leading to admission at the hospital and the development of convulsions were unrelated to the prognosis . Those who succumbed or suffered sequelae were all infants under 1 year of age . All cases were caused by genus Haemophilus.

Kansenshogaku Zasshi, 2004 Jan, 78(1), 40 - 5
{Antimicrobial susceptibilities of Streptococcus pneumoniae and Haemophilus influenzae isolated from children with meningitis}; Sakata H; Between April 2001 and March 2003, we studied minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of 7 strains of Streptococcus pneumoniae and 8 strains of Haemophilus influenzae isolated from children with meningitis . The age range of the patients was from 4 months to 5 years . Susceptibilities of ampicillin (ABPC), cefotaxime (CTX), panipenem (PAPM), and vancomycin (VCM) in S . pneumoniae and those of ABPC, CTX, ceftriaxone (CTRX), and meropenem (MEPM) in H . influenzae were measured . MICs of ABPC, CTX, PAPM, and VCM to S . pneumoniae were < or = 0.06-2, < or = 0.06-0.5, < or = 0.06, and 0.25-0.5 microgram/ml and MICs of ABPC, CTX, CTRX, and MEPM to H . influenzae were 0.12-64, < or = 0.06-0.5, < or = 0.06-0.12, and < or = 0.06-0.25 microgram/ml, respectively . In 5 of all strains, difference between MIC and MBC to ABPC was observed . Four strains out of them had mutations of penicillin binding protein genes measured by PCR methods.

Infect Immun, 2004 May, 72(5), 3002 - 10
Partial analysis of the genomes of two nontypeable Haemophilus influenzae otitis media isolates; Munson RS Jr et al.; In 1995, The Institute for Genomic Research completed the genomic sequence of a rough derivative of Haemophilus influenzae serotype d, strain KW20 . This sequence, though extremely useful in understanding the basic biology of H . influenzae, has yet to provide significant insight into our understanding of disease caused by nontypeable H . influenzae (NTHI), because serotype d strains are not generally pathogens . In contrast, NTHI strains are frequently mucosal pathogens and are the primary pathogens of chronic otitis media as well as a significant cause of acute otitis media in children . Thus, it is of great importance to further understand their biology . We used a DNA-based microarray approach to identify genes present in a clinical isolate of NTHI that were absent from strain Rd . We also sequenced the genome of a second NTHI isolate from a child with chronic otitis media to threefold coverage and then used an array of bioinformatics tools to identify genes present in this NTHI strain but absent from strain Rd . These methods were complementary in approach and results . We identified, in both strains, homologues of H . influenzae lav, an autotransported protein of unknown function; tnaA, which encodes tryptophanase; as well as a homologue of Pasteurella multocida tsaA, which encodes an alkyl peroxidase that may play a role in protection against reactive oxygen species . We also identified a number of putative restriction-modification systems, bacteriophage genes and transposon-related genes . These data provide new insight that complements and extends our ongoing analysis of NTHI virulence determinants.

Arch Dis Child Fetal Neonatal Ed, 2004 May, 89(3), F269 - 71
Responses to a fourth dose of Haemophilus influenzae type B conjugate vaccine in early life; Slack MH et al.; OBJECTIVE: To describe the immune response of preterm infants, with a reduced response to primary Haemophilus influenzae type B (Hib) immunisation, to a fourth dose of Hib conjugate vaccine given in early life . DESIGN: Prospective observational study . SETTING: Five Wessex Neonatal Units . PATIENTS: Infants born at < 32 weeks and immunised with three doses of combined acellular pertussis-Hib vaccine, with a Hib IgG geometric mean concentration (GMC) < 1.0 microg/ml after these primary immunisations . INTERVENTIONS: An additional fourth dose of Hib conjugate vaccine given before 1 year of age . Blood taken to assess Hib IgG concentration and avidity after immunisation . MAIN OUTCOME MEASURES: Hib IgG GMC and avidity index . RESULTS: Ninety six infants (mean gestational age at birth 29.1 weeks) received a fourth dose of Hib at a mean age of 7.8 months . Hib IgG GMC after the primary immunisations was 0.17 microg/ml (95% confidence interval (CI) 0.14 to 0.20) rising to 4.68 microg/ml (95% CI 3.36 to 6.57) after the fourth dose (p < 0.0001) . The IgG response to the fourth dose correlated positively with the response after the primary immunisations (p < 0.001) . Hib IgG geometric mean avidity index (GMAI) after the primary immunisations was 30.87 (95% CI 20.40 to 46.73) . This increased to 124.73 (95% CI 109.93 to 141.51) after the fourth dose (p < 0.0001) . CONCLUSION: Preterm infants with very low IgG responses to Hib after primary immunisations with a combined acellular pertussis-Hib vaccine mount a good response to a fourth dose of Hib . This study suggests that all infants will benefit from a fourth dose of Hib, regardless of the age at which it is given.

Expert Opin Pharmacother, 2004 Apr, 5(4), 855 - 63
Meningococcal conjugate vaccines; Zimmer SM et al.; Neisseria meningitidis is a leading cause of bacterial meningitis and sepsis in the US, Europe and in many other parts of the world, including parts of sub-Saharan Africa (known as the African 'meningitis belt') . There are > 500000 cases of meningococcal disease annually with an estimated death toll of 135000 worldwide . Approximately 10 - 15 % of survivors experience significant morbidity in the form of neurological sequelae, including hearing loss, speech disorders, loss of limbs, mental retardation and paralysis . Disease is usually caused by N . meningitidis serogroups A, B, C, Y or W-135 . Prevention of meningococcal disease includes isolation, chemoprophylaxis and vaccination with available polysaccharide vaccines . However, the polysaccharide meningococcal vaccines (i.e., A and C; A, C and W-135; or A, C, Y and W-135) initially developed in the 1970s are generally poorly immunogenic in children or require repeated doses and do not produce long-lasting immunity . Conjugate vaccine technology has been very successfully used in childhood vaccines for the prevention of other bacterial meningitis pathogens, including vaccines against Haemophilus influenzae serotype b (Hib) and more recently, the seven- and nine-valent conjugate pneumococcal vaccines . Newly released meningococcal conjugate vaccines against N . meningitidis serogroup C have been highly efficacious in young children and adolescents, with minimal side effects . Conjugate vaccines targeting other important meningococcal serogroups (e.g., N . meningitidis serogroup A, responsible for the large pandemic outbreaks and the majority of disease in sub-Saharan Africa and serogroups Y and W-135) are under development and together with the serogroup C conjugates, have the potential to significantly impact worldwide sporadic and epidemic meningococcal disease . The search for an effective serogroup B meningococcal vaccine remains elusive . This manuscript reviews the conjugate meningococcal vaccines and their potential for meningococcal disease prevention.

Klin Mikrobiol Infekc Lek, 2004 Feb, 10(1), 22 - 4
{Haemophilus influenzae meningitis in a child with complete immunisation}; Zjevikova A et al.; Invasive Haemophilus influenzae b (Hib) diseases are rarely observed in children with vaccination against Hib . Immunisation of infants against Hib was introduced in the Czech Republic in 2001 . The first case of meningitis with isolation of Hib in child immunised with Hib vaccine since starting routine vaccination is described . Clinical course of meningitis was mild and was identical with previously manifestation of Hib meningitis.

Nucleic Acids Res, 2004 Apr 19, 32(7), 2147 - 57 Print 2004.
Operon prediction by comparative genomics: an application to the Synechococcus sp . WH8102 genome; Chen X et al.; We present a computational method for operon prediction based on a comparative genomics approach . A group of consecutive genes is considered as a candidate operon if both their gene sequences and functions are conserved across several phylogenetically related genomes . In addition, various supporting data for operons are also collected through the application of public domain computer programs, and used in our prediction method . These include the prediction of conserved gene functions, promoter motifs and terminators . An apparent advantage of our approach over other operon prediction methods is that it does not require many experimental data (such as gene expression data and pathway data) as input . This feature makes it applicable to many newly sequenced genomes that do not have extensive experimental information . In order to validate our prediction, we have tested the method on Escherichia coli K12, in which operon structures have been extensively studied, through a comparative analysis against Haemophilus influenzae Rd and Salmonella typhimurium LT2 . Our method successfully predicted most of the 237 known operons . After this initial validation, we then applied the method to a newly sequenced and annotated microbial genome, Synechococcus sp . WH8102, through a comparative genome analysis with two other cyanobacterial genomes, Prochlorococcus marinus sp . MED4 and P.marinus sp . MIT9313 . Our results are consistent with previously reported results and statistics on operons in the literature.

Chang Gung Med J, 2004 Feb, 27(2), 122 - 8
Clinical analysis of bronchiectasis in Taiwanese children; Lai SH et al.; BACKGROUND: The clinical features and etiology of bronchiectasis have rarely been described in non-Caucasian populations, of whom the prevalence of cystic fibrosis is low . In this report, we studied the clinical features of bronchiectasis in Taiwanese children . METHODS: Using a retrospective chart review, 29 cases of bronchiectasis were diagnosed from 1991 through 2001 . For each case, the diagnosis was confirmed using high-resolution computed tomography . Medical records were analyzed for demographic data, clinical presentation, spirometric data, and microbial isolation . Radiographic findings were reviewed, and possible causes of bronchiectasis were also identified . RESULTS: There were 17 girls and 12 boys enrolled . Persistent cough, daily sputum production, and hemoptysis were common presenting symptoms . Crackles and wheezing were the most frequent findings during the physical examination . Previous lower airway infection, asthma, and primary immunodeficiency were the most common, but 31.0% of the cases had unknown causes . Dependent lobes were involved more frequently . Simultaneous sinusitis was noted in 70.6% of the cases . Spirometry showed mild airway obstruction in most of the cases . Most specimens (52.2%) from lower airway secretions yielded bacterial pathogens, most commonly Pseudomonas aeruginosa, Haemophilus influenzae and Streptococcus pneumoniae . CONCLUSION: Continuing post-infectious inflammatory changes remains the most important cause of bronchiectasis in a non-Caucasian pediatric population in Northern Taiwan in the 1990s . More than two thirds of these patients had underlying predisposing factors including asthma, immunodeficiency, and swallowing dysfunction . Chronic productive purulent respiratory secretions and persistent crackles should raise the possibility of bronchiectasis in children even in Taiwan where the incidence of cystic fibrosis is low.

Laryngoscope, 2004 Mar, 114(3), 472 - 85
Otitis media: treatment with intranasal aerosolized surfactant; Chandrasekhar SS et al.; OBJECTIVES/HYPOTHESIS: The objective was to determine the effect of intranasal surfactant alone and with other medications administered by metered dose inhaler aerosol on the function of the eustachian tube and on the resolution of experimentally induced otitis media with effusion (OME) and acute otitis media (AOM) . STUDY DESIGN: Randomized, experimental, controlled animal studies . METHODS: Previously unreported (experiment 4) as well as published (experiments 1-3) data were detailed so that the reader could understand the continuum of information leading to the conclusions . In experiment 1, after a live-animal technique of measuring eustachian tube passive opening pressure was developed and validated, eustachian tube passive opening pressure was determined in 61 adult gerbils and 34 mice at baseline and 5 and 10 minutes after delivery of aerosolized intranasal metered dose inhaler surfactant . In experiments 2 and 3 (Klebsiella pneumoniae), lipopolysaccharide-induced OME was developed in gerbils . Thirty-five animals were randomly assigned to control, placebo, surfactant, surfactant with betamethasone, and surfactant with phenylephrine groups . Seventy animals were randomly assigned to control, placebo once daily (QD) and twice daily (BID), surfactant QD and BID, surfactant with betamethasone QD and BID, and surfactant with phenylephrine QD and BID groups . Intranasal aerosolized MDI medications were administered from postinfection day 2 until the effusion resolved . Otomicroscopy and tympanometry were performed on alternate days for 30 days . In experiment 4, AOM was developed in 39 chinchillas via transbullar injection of nontypeable Haemophilus influenzae on day 1 . Thirteen animals each received placebo BID or surfactant BID, beginning on day 1 . Thirteen animals received surfactant BID beginning on day 3 . All administrations were continued for 10 days . Examinations were performed on seven occasions until day 27 . Appropriate statistical measurements were employed, including one- and two-way ANOVA, strength-of-association measure (omega) calculation, chi, and Newman-Keuls post hoc multiple comparison tests . Significance was set as P value of less than.05 . RESULTS: In experiment 1, a significant reduction in passive opening pressure was seen in both 5- and 10-minute postsurfactant measurements . Propellant alone was not effective . In experiments 2 and 3, OME resolved after an average period of 16 to 16.5 days in control, placebo QD and BID, and surfactant with phenylephrine QD groups . A significant decrease in OME days was seen in the surfactant QD (10.57 d) and BID (8.57 d), and surfactant with betamethasone QD (8.57 d) and BID (6.3 d) groups . A significant increase was seen in the phenylephrine BID group (18.67 d) . In experiment 4, tympanometry was normal or near-normal in 62% and 48% of treated ears and in only 24% of placebo ears on day 12 . Sixty-seven percent of placebo ears were culture positive at day 27, compared with 10% and 16% in surfactant groups 1 and 2 . Seventy-five percent of untreated animals developed severe labyrinthitis, compared with 15% in groups 1 and 2 . On day 27, 58% of placebo group middle ears had fluid, whereas 61% and 62% of ears in groups 1 and 2, respectively, were dry . These findings were significant . CONCLUSION: Intranasal application of aerosolized metered dose inhaler surfactant alone or with steroid reduced eustachian tube passive opening pressure in normal animals and duration of effusion in animals with experimental OME . Intranasal surfactant reduced the severity and duration of middle ear infection in AOM in this animal model.

J Vet Med A Physiol Pathol Clin Med, 2004 Feb, 51(1), 10 - 4
Immunohistopathological findings in the lungs of calves naturally infected with Mycoplasma bovis; Khodakaram-Tafti A et al.; Histology and immunohistochemistry were used to analyse the lesions and distribution of Mycoplasma bovis antigen in the lungs of 18 naturally infected calves . Microscopic examination of pneumonic lungs revealed two distinct patterns of necrosis and inflammation . The first pattern was observed in six of 18 (33.3%) calves in which microscopic lesions were characterized by large irregular areas of coagulative necrosis surrounded by a dense zone of degenerated neutrophils . Moderate amounts of mycoplasmal antigen were in the centre and periphery of these necrotic foci and, to a lesser extent, in mononuclear cells of the peribronchial lymphoid tissue . The second pattern was observed in 18 of 18 (100%) calves and consisted of rounded foci of caseous necrosis composed by granular eosinophilic material surrounded by a rim of granulation tissue . Large amounts of M . bovis antigen were detected in the centre and periphery of these necrotic foci and, to a lesser extent, in the peribronchial lymphoid tissue, and alveolar and interstitial macrophages . It was concluded that both caseous and coagulative necrosis of the lung parenchyma was primarily caused by M . bovis . Infection with M . bovis should be suspected in bovine necrotic bronchopneumonia, particularly in cases in which the pulmonary necrosis is part of a pyogranulomatous inflammation centred around airways . The pattern of caseous necrosis with pyogranulomatous inflammation is characteristic of M . bovis infection while the pattern of coagulative necrosis is similar to and must be differentiated from Mannheimia haemolytica and Haemophilus somnus infection.

Int J Pediatr Otorhinolaryngol, 2004 May, 68(5), 545 - 50
Cochlear implantation and meningitis; Callanan V et al.; OBJECTIVE: Study clinical presentation, diagnosis and treatment of cochlear implant patients diagnosed with bacterial meningitis . BACKGROUND: Cochlear implantation in children is necessary for the optimal development of speech in the developing child diagnosed with profound sensorineural hearing loss . Approximately 60,000 devices have been inserted in adults and children worldwide to date . SETTING: The Department of Pediatric Otolaryngology of a tertiary care children's hospital . METHODS: All patients undergoing cochlear implantation from April 1997 were identified . Patients diagnosed with bacterial meningitis after implantation were selected for study . RESULTS: Of 30 children, two developed bacterial meningitis after cochlear implantation . One patient developed Streptococcus pneumoniae meningitis . One patient developed nontypable Haemophilus influenzae bacterial meningitis . Both patients made a complete recovery . CONCLUSIONS: Prompt diagnosis and treatment of meningitis is essential to optimize outcome.

Int J Antimicrob Agents, 2004 Apr, 23(4), 356 - 61
Antimicrobial resistance of Streptococcus pneumoniae and Haemophilus influenzae in Sao Paulo, Brazil from 1996 to 2000; Koeth LM et al.; This study was undertaken to assess the in vitro activity of several antimicrobial agents against Brazilian isolates of Streptococcus pneumoniae and Haemophilus influenzae from 1996 to 2000 . The antibiotics used were penicillin, amoxicillin/clavulanic acid (A/C), ampicillin, amoxicillin, cefaclor, cefdinir, cefixime, cefprozil, ceftriaxone, cefuroxime, azithromycin, clarithromycin, erythromycin, ciprofloxacin, levofloxacin, ofloxacin, chloramphenicol, clindamycin, doxycycline and trimethoprim/sulphamethoxazole (T/S) . MICs were determined by the National Committee for Clinical Laboratory Standards (NCCLS) method and interpreted using NCCLS and PK/PD breakpoints . For S . pneumoniae 80.0% were penicillin susceptible, 18.3% intermediate, 1.7% resistant; most active agents were amoxicillin, A/C, ceftriaxone and levofloxacin; T/S was the least active agent . Beta-lactamase was produced by 13.7% of H . influenzae . All were susceptible to A/C, cefdinir, cefixime, ceftriaxone and quinolones . The least active agents were T/S and macrolides.

Cent Eur J Public Health, 2003 Dec, 11 Suppl, S25 - 30
The 2001 serological survey in the Czech Republic--Hib invasive disease Haemophilus influenzae b; Lebedova V et al.; Dynamics in total antibody levels against Haemophilus influenzae b (Hib) among the Czech population were monitored for the first time within the 2001 serological survey . In total, 2,479 non-vaccinee sera and 65 vaccinee sera from the population of age groups 1-39 years were tested . The total anti-Hib antibody levels determined by means of ELISA were higher than 0 . 15 microg/ml for all of the sera tested . Levels of 1 microg/ml or higher were recorded for 97% and 99% of vaccinee and non-vaccinee sera, respectively . The non-vaccinees showed the highest rates of levels below 1 microg/ml in the age groups of 1-, 2-, 6- and 12-year-olds (4%, 7%, 3% and 3%, respectively) . Only one child aged 1 year had a level below 1 microg/ml among the vaccinees . Levels of 2 microg/ml and higher were found in 89% and 91% of non-vaccinee and vaccinee sera, respectively . The non-vaccinees showed the highest rates of levels below 2 microg/ml in the age groups of 1-, 2-, 3-, 4- and 6-year-olds (46%, 37%, 22%, 20% and 18%, respectively), the other age groups were characterized by lower rates (2-13%) . Levels below 2 microg/ml were found in 1-, 2-, and 4-year-old vaccinees (25%, 4% and 9%, respectively) . In conclusion, it can be stated that the immunity of most of the Czech population is very good . Nevertheless, children under 6 years of age are at highest risk as also confirmed by the programme of Hib invasive disease surveillance . As many as 94 cases of Hib invasive diseases were recorded in the Czech Republic in 2001, 82 (87%) of them in children aged between 0 to 6 years, with only sporadic cases being detected in the other age groups.

J Chemother, 2004 Feb, 16(1), 30 - 7
Effect of moxifloxacin on bacterial pathogenicity factors in comparison with amoxicillin, clarithromycin and ceftriaxone; Drago L et al.; Moxifloxacin is a recent fluoroquinolone with an antibacterial spectrum encompassing both aerobic Gram-negative and Gram-positive strains, as well as anaerobic bacteria . In this study the activity of moxifloxacin against Streptococcus pneumoniae, Staphylococcus aureus, Moraxella catarrhalis, Haemophilus influenzae, Escherichia coli, Proteus mirabilis and Pseudomonas aeruginosa, and effects of subinhibitory concentrations on bacterial morphology and adhesion properties were compared with those of amoxicillin, clarithromycin and ceftriaxone . The in vitro activity of moxifloxacin against Gram-positive and Gram-negative pathogens was equal to or better than that of comparators . Subinhibitory concentrations of moxifloxacin significantly affected bacterial morphology of S . pneumoniae, M . catarrhalis, H . influenzae and P . aeruginosa, leading to formation of spherical forms and filaments . Moreover, bacterial adhesion to buccal cells and fibroblasts was reduced after treatment with 1/4 and 1/8 X MIC of moxifloxacin . In conclusion, subinhibitory concentrations of moxifloxacin remarkably interfere with some bacterial pathogenic factors, thereby contributing to its antimicrobial activity.

J Chemother, 2004 Feb, 16(1), 23 - 9
In vitro studies on the impact of human serum on the antibacterial effect of faropenem; MacGowan A et al.; The interaction between faropenem and serum in determining antibacterial effect was studied using three target pathogens and two different in vitro methodologies . Strains of Staphylococcus aureus, Streptococcus pneumoniae, capsulate and non capsulate Haemophilus influenzae, all faropenem MIC 0.12 mg/L, were tested in a range of pharmacologically realistic faropenem concentrations with various proportions of serum up to 75% . Using simulated serum bactericidal titres the presence of human serum reduced the activity of faropenem against S . pneumoniae as did heat-treated serum for non-capsulate H . influenzae . Serum on its own was bactericidal against H . influenzae probably due to the presence of complement . The antibacterial effects of combinations of faropenem and serum was assessed in time-kill curves by calculation of the area-under-the bacterial-kill-curve (AUBKC) . This was then related to faropenem concentration and the proportion of serum using three-dimensional plots . Serum on its own was mildly inhibitory of the growth of S . aureus, supported improved growth of S . pneumoniae at some proportions and was rapidly bactericidal to H . influenzae, especially the non-capsulate strains . Faropenem had a marked antibacterial effect against all three species in the range 0-2.5 mg/L . Increasing the faropenem concentration from 2.5-10 mg/L produced little or no additional effect . The combination of serum and faropenem had little impact on the antibacterial effect against S . pneumoniae and S . aureus but free drug concentrations were likely to be greater than the MIC in all the combinations used . Against capsulate H . influenzae the effect of serum and faropenem was broadly equivalent while against non-capsulate strains the activity of serum was so great it is difficult to assess the impact of faropenem alone . The interaction between serum and antibiotic in determining antibacterial effect is complex and critically dependent on the proportions of serum and drug concentrations chosen . Three-dimensional plots offer a tool to visualise these complex relationships which may be species specific.

Int J Epidemiol, 2004 Feb, 33(1), 173 - 81
Effectiveness of Haemophilus influenzae b conjugate vaccine on childhood pneumonia: a case-control study in Brazil; de Andrade AL et al.; BACKGROUND: The conjugate Haemophilus influenzae type b (Hib) vaccine has been highly efficacious in reducing type b H . influenzae meningitis . However, information is limited about its impact on childhood pneumonia after vaccine introduction into the routine programme . This study evaluated the effectiveness of Hib conjugate vaccine in the reduction of community-acquired pneumonia among infants in Central Brazil . METHODS: A matched case-control study was built into an ongoing prospective population-based surveillance of pneumonia, enrolling 1293 participants between May 2000 and August 2001 . Cases (n = 431) were children <or=2 years old hospitalized with radiologically definite pneumonia according to the World Health Organization standard criteria for the interpretation of radiographs for the diagnosis of pneumonia . Two controls (n = 862) without previous hospitalization for pneumonia were identified among children from the same neighbourhood and matched to cases by age stratum (+/-4 months) . The Hib vaccination effectiveness was estimated as 1 minus odds ratio (OR) . RESULTS: The greatest risk factor for pneumonia among children <2 years of age was day-care centre attendance (P < 0.001) . Of the study participants, 83.3% (1072/1287) were classified as vaccinated according to immunization card and 16.7% (215/1287) were considered unvaccinated . By using conditional logistic regression the vaccine effectiveness was estimated as 31.0% (95% CI: -9.0%, 57.0%) after adjusting for sex, previous flu-like illnesses, day-care attendance, smokers at home, house ownership, mother's education, and age as continuous variable . CONCLUSION: Under programme conditions the effectiveness of Hib conjugate vaccine in infants with radiologically confirmed pneumonia was 31% (95% CI: -9%, 57%) showing the potential benefit of Hib immunization in the prevention of likely non-bacteraemic pneumonia.

Curr Opin Pulm Med, 2004 May, 10(3), 176 - 82
Pneumonia in HIV-infected and HIV-uninfected children in developing countries: epidemiology, clinical features, and management; Zar HJ; PURPOSE OF REVIEW: Pneumonia is a leading cause of illness and death in children younger than 5 years in developing countries, accounting for approximately 20% of childhood deaths . The HIV epidemic has sharply increased the incidence, severity, and mortality of childhood pneumonia in the developing world, particularly in sub-Saharan Africa . This article reviews recent findings on the epidemiology, clinical features, and management of HIV-infected and -uninfected children with pneumonia in developing countries . RECENT FINDINGS: Bacterial infection remains a major cause of pneumonia mortality; in HIV-infected children, a broader spectrum of pathogens including gram-negative infections and Pneumocystis jiroveci occurs . Mycobacterium tuberculosis is an important cause of acute pneumonia among children from high tuberculosis prevalence areas . Use of case management guidelines substantially reduces neonatal, infant, and under-5 mortality and pneumonia-specific mortality in developing countries . New advances in therapy include the use of short-course antibiotics and high-dose amoxicillin twice daily for ambulatory treatment of HIV-negative children with pneumonia . New preventive interventions include the development of conjugate vaccines against Streptococcus pneumoniae and Haemophilus influenzae, but these are not widely affordable nor available in developing countries . Despite a lower efficacy in HIV-infected children, these vaccines still protect against disease in a significant proportion of children . Available preventive interventions including micronutrient supplementation with zinc and vitamin A, and immunization as contained in the WHO Expanded Program of Immunization can substantially reduce the burden of childhood pneumonia . SUMMARY: Urgent measures to implement existing available, effective interventions for prevention and treatment of childhood pneumonia and achieve high coverage rates in developing countries are needed.

J Clin Microbiol, 2004 Apr, 42(4), 1450 - 9
Dynamics of long-term colonization of respiratory tract by Haemophilus influenzae in cystic fibrosis patients shows a marked increase in hypermutable strains; Roman F et al.; The persistence and variability of 188 Haemophilus influenzae isolates in respiratory tract of 30 cystic fibrosis (CF) patients over the course of 7 years was studied . Antibiotic susceptibility testing, DNA fingerprinting, and analysis of outer membrane protein profiles were performed on all isolates . A total of 115 distinct pulsed-field gel electrophoresis profiles were identified . Ninety percent of patients were cocolonized with two or more clones over the studied period . A third of the patients were cross-colonized with one or two H . influenzae strains; 11% of the clones persisted for 3 or more months . Biotype, outer membrane protein profiles, and resistance profiles showed variation along the studied period, even in persisting clones . Four isolates (2.1%) recovered from 3 patients were type f capsulate, with three of them belonging to the same clone . beta-Lactamase production was detected in 23.9% of isolates while 7% of the beta-lactamase-negative isolates presented diminished susceptibility to ampicillin (beta-lactamase-negative ampicillin resistance phenotype) . Remarkably, 21.3% of the H . influenzae isolates presented decreased susceptibility to ciprofloxacin, which was mainly observed in persisting clones . Of the H . influenzae isolates from CF patients, 18 (14.5%) were found to be hypermutable in comparison with 1 (1.4%) from non-CF patients (P < 0.0001) . Ten patients (33.3%) were colonized by hypermutable strains over the study period . A multiresistance phenotype and long-term clonal persistence were significantly associated in some cases for up to 7 years . These results suggest that H . influenzae bronchial colonization in CF patients is a dynamic process, but better-adapted clones can persist for long periods of time.

N Engl J Med, 2004 Apr 1, 350(14), 1398 - 404
Childhood vaccination and type 1 diabetes; Hviid A et al.; BACKGROUND: A link between childhood vaccinations and the development of type 1 diabetes has been proposed . METHODS: We evaluated a cohort comprising all children born in Denmark from January 1, 1990, through December 31, 2000, for whom detailed information on vaccinations and type 1 diabetes was available . Using Poisson regression models, we estimated rate ratios according to vaccination status, including the trend associated with the number of doses, among all children and in a subgroup of children who had siblings with type 1 diabetes . Given recent claims of clustering of cases of diabetes two to four years after vaccination, we also estimated rate ratios during the period after vaccination . RESULTS: Type 1 diabetes was diagnosed in 681 children during 4,720,517 person-years of follow-up . The rate ratio for type 1 diabetes among children who received at least one dose of vaccine, as compared with unvaccinated children, was 0.91 (95 percent confidence interval, 0.74 to 1.12) for Haemophilus influenzae type b vaccine; 1.02 (95 percent confidence interval, 0.75 to 1.37) for diphtheria, tetanus, and inactivated poliovirus vaccine; 0.96 (95 percent confidence interval, 0.71 to 1.30) for diphtheria, tetanus, acellular pertussis, and inactivated poliovirus vaccine; 1.06 (95 percent confidence interval, 0.80 to 1.40) for whole-cell pertussis vaccine; 1.14 (95 percent confidence interval, 0.90 to 1.45) for measles, mumps, and rubella vaccine; and 1.08 (95 percent confidence interval, 0.74 to 1.57) for oral poliovirus vaccine . The development of type 1 diabetes in genetically predisposed children (defined as those who had siblings with type 1 diabetes) was not significantly associated with vaccination . Furthermore, there was no evidence of any clustering of cases two to four years after vaccination with any vaccine . CONCLUSIONS: These results do not support a causal relation between childhood vaccination and type 1 diabetes .

Proc Natl Acad Sci U S A, 2004 Mar 30, 101(13), 4513 - 8 Epub 2004 Mar 19.
Evolutionary stability of DNA uptake signal sequences in the Pasteurellaceae; Bakkali M et al.; The DNA-uptake signal sequence (USS) of the bacterium Haemophilus influenzae is highly over-represented in its genome (1,471 copies of the core sequence AAGTGCGGT), and DNA fragments containing USS are preferentially taken up by competent cells . Because this bias favors uptake of conspecific DNA, USSs are often considered a kind of mate recognition system in bacteria, acting as species-specific barriers against uptake of unrelated DNA . However, the H . influenzae USS is highly over-represented in the genomes of three otherwise-divergent Pasteurellaceae species (Pasteurella multocida, Haemophilus somnus, and Actinobacillus actinomycetemcomitans, 927, 1,205, and 1,760 copies, respectively), suggesting that USSs do not always limit exchange . USSs in all these genomes are mainly in coding regions and show no orientation bias around the chromosome, weakening proposed USS functions in transcription termination and chromosome replication . Alignment of homologous genes was used to determine evolutionary relationships between individual USSs . Most H . influenzae USSs were found to have perfect or imperfect homologs (USS at the same location) in at least one other species, and most USSs in the other species had perfect or imperfect homologs in H . influenzae . These homologies suggest that the use of a common USS is due to inheritance of the USS-based uptake system from a common ancestor of the Pasteurellaceae, and it indicates that individual USSs can be evolutionarily stable elements of their genomes . The pattern is consistent with a molecular drive model of USS evolution, with new USSs arising by mutation and preferentially spread to new genomes by the biased DNA-uptake system.

BMJ . 2004 Apr 3;328(7443):791 . Epub 2004 Mar 16.
Three day versus five day treatment with amoxicillin for non-severe pneumonia in young children: a multicentre randomised controlled trial; Agarwal G et al.; OBJECTIVE: To assess the efficacy of three days versus five days of treatment with oral amoxicillin for curing non-severe pneumonia in children . DESIGN: Randomised, double blind, placebo controlled multicentre trial . SETTING: Outpatient departments of seven referral hospitals in India . PARTICIPANTS: 2188 children aged 2-59 months, 1095 given three days of treatment and 1093 given five days . INTERVENTION: Oral amoxicillin 31-54 mg/kg/day in three divided doses . MAIN OUTCOME MEASURES: Treatment failure: defined as development of chest indrawing, convulsions, drowsiness, or inability to drink at any time; respiratory rate above age specific cut points on day 3 or later; or oxygen saturation by pulse oximetry < 90% on day 3 . RESULTS: The clinical cure rates with three days and five days of treatment were 89.5% and 89.9%, respectively (absolute difference 0.4 (95% confidence interval--2.1 to 3.0)) . Adherence to treatment regimen was 94% and 85% for three day and five day treatments, respectively . Loss to follow up was 5.4% by day 5 . There were no deaths, 41 hospitalisations, and 36 minor adverse reactions . There were 225 (10.3%) clinical failures and 106 (5.3%) relapses, and rates were similar in both treatments . At enrollment, 513 (23.4%) children tested positive for respiratory syncytial virus, and Streptococcus pneumoniae and Haemophilus influenzae were isolated from the nasopharynx in 878 (40.4%) and 496 (22.8%) children, respectively . Clinical failure was associated with isolation of respiratory syncytial virus (adjusted odds ratio 1.95 (95% confidence interval 1.0 to 3.8)), excess respiratory rate of > 10 breaths/minute (2.89 (1.83 to 4.55)), and non-adherence with treatment at day 5 (11.57 (7.4 to 18.0)) . CONCLUSIONS: Treatment with oral amoxicillin for three days was as effective as for five days in children with non-severe pneumonia.

J Infect, 2004 May, 48(4), 320 - 9
Risk factors for invasive disease among children in Spain; Pereiro I et al.; OBJECTIVE: To identify the risk factors related to invasive disease caused by Haemophilus influenzae type b (Hib), Neisseria meningitidis, and Streptococcus pneumoniae . METHODS: Case-control study . All hospitals of the region of Valencia in Spain (covering 95% of the population of Valencia) . The patients are children aged less than 15 years in whom Hib, N . meningitidis or S . pneumoniae are isolated from normally sterile sites . RESULTS: From 1995 to 1998, 275 cases of invasive disease were analysed, and 243 hospital controls were selected in the month after the onset of the case . The paediatrician completed a survey administered to the relatives at the time of admission . The risk factors related to invasive disease by Hib were exposure to tobacco smoke (number of smokers, adjusted OR (aOR) 1.74, 95% confidence interval (CI) 1.02-2.96) and living with more than four people (aOR 3.72, 95% CI 1.3-3.7) . For N . meningitidis, there is a dose-response relationship; if more than 60 cigarettes are smoked daily at home, the aOR is 3.61 (95% CI 1.04-12.57) . If there are more than four people living in the household, aOR 1.69 (95% CI 1.01-2.85) . In children under two years of age, having siblings less than 15 years of age (OR 1.76, 95% CI 0.75-4.17) and attending a day nursery represents a risk for suffering invasive pneumococcal disease (aOR 4.21, 95% CI 1.28-13.83) . CONCLUSIONS: Among the variables tested, the modifiable risk factor is smoking; if smoking was reduced at home, the number of cases of invasive disease could be reduced in children, mainly in those under 5 years of age . Identification and vaccination of these risk groups would significantly reduce these diseases.

Vaccine, 2004 Mar 29, 22(11-12), 1415 - 22
Antibody persistence against diphtheria, tetanus, pertussis, poliomyelitis and Haemophilus influenzae type b (Hib) in 5-6-year-old children after primary vaccination and first booster with a pentavalent combined acellular pertussis vaccine: immunogenicity and tolerance of a tetravalent combined acellular pertussis vaccine given as a second booster; Mallet E et al.; The main objective of this study was to assess in 5-6-year-old French children (n=234) the persistence of antibodies induced by a primary series vaccination (at 2-4 months of age) and a first booster (at 12-16 months of age) with a pentavalent two-component acellular pertussis combined vaccine (DTacP-IPV-Hib; Pentavac) . The second objective was to evaluate in these 5-6-year-old French children the safety and the immunogenicity of a tetravalent acellular pertussis combined vaccine (DTacP-IPV; Tetravac) given as second booster . RESULTS: Seroprotective antibody levels against diphtheria, tetanus, types 1-3 poliomyelitis and PRP were maintained 4-5 years after primary-vaccination and first booster with Pentavac . As expected, anti-PT antibodies levels were low, suggesting that children were not colonised by Bordetella pertussis . The second booster with Tetravac was well tolerated and elicited a strong booster response for all antigens . CONCLUSION: acellular pertussis combined vaccine, used in primary-vaccination, could be considered as having the same priming effect and the same efficacy as whole cell pertussis vaccine.

Vaccine, 2004 Mar 29, 22(11-12), 1343 - 57
A liquid hexavalent combined vaccine against diphtheria, tetanus, pertussis, poliomyelitis, Haemophilus influenzae type B and hepatitis B: review of immunogenicity and safety; Mallet E et al.; To reduce the number of injections needed to comply with paediatric vaccination requirements, a liquid, hexavalent vaccine (DTaP-IPV-PRP-T-HBs; Hexavac; Aventis Pasteur MSD) has been developed for primary and booster vaccination of infants and toddlers . In extensive clinical studies, Hexavac has been shown to be highly immunogenic . Seroconversion or seroprotective titres of antibodies against all antigens were achieved in the majority of infants following a primary series of three doses administered at 1-2-month intervals from 2 months of age . Hexavac also induced immunologic memory, as evidenced by the anamnestic response to booster vaccination at 12-18 months of age . These responses were comparable with those seen following concomitant administration of Pentavac (DTaP-IPV//PRP-T) and monovalent hepatitis B vaccine (H-B-Vax II), and were also within the ranges observed for other relevant licensed vaccines . Clinical studies comparing the immunogenicity of Hexavac administered at either 2, 3 and 4 months or 2, 4 and 6 months demonstrated that it can be used by either vaccination schedule . A further study also supported the use of primary doses of Hexavac at 3 and 5 months with a booster at 12 months of age . Hexavac demonstrated a good reactogenicity and tolerability profile . The most frequently reported adverse events after both primary and booster doses were local reactions of redness and swelling/induration and a systemic response of mild fever, irrespective of the vaccine used for priming . Hexavac provided immunity against six important childhood diseases with a single injection at each visit.

FEMS Microbiol Lett, 2004 Apr 15, 233(2), 277 - 81
Harnessing natural transformation in Actinobacillus pleuropneumoniae: a simple method for allelic replacements; Bosse JT et al.; We have investigated the use of a natural transformation protocol to introduce mutations into Actinobacillus pleuropneumoniae serotypes 1 and 5b . For both strains tested, we recovered 1 in 10(8) transformants during culture in rich medium, a result that was independent of the growth phase . This low frequency of transformation of A . pleuropneumoniae did not increase when bacteria were grown under conditions known to be optimal for induction of competence in Haemophilus influenzae . Using linearised plasmid DNA containing a kanamycin cassette inserted into the sodC gene of A . pleuropneumoniae serotype 1, we showed that natural transformation can be used as a simple method for introducing allele replacements into this bacterium, and can be used to transfer mutations from one serotype to another.

Pediatrics, 2004 Apr, 113(4), e296 - 302
Immunization coverage levels among 19- to 35-month-old children in 4 diverse, medically underserved areas of the United States; Rosenthal J et al.; BACKGROUND: The National Immunization Survey demonstrates that national immunization coverage in 2002 remained near the all-time highs achieved in 2000 . However, that survey cannot detect whether coverage is uniformly high within relatively small areas or populations . The measles resurgence in the early 1990s revealed that coverage was low in some areas, particularly among inner-city children from racial and ethnic minority groups . Today, identifying areas with low childhood-vaccination coverage remains important, particularly if these areas are at risk for the introduction of disease . In 1995, the Centers for Disease Control and Prevention launched a congressionally mandated demonstrated project now called the Childhood Immunization Demonstration project of Community Health Networks . This mandate specified an assessment to determine whether a network of primary care providers affiliated with university teaching hospitals could assume a public health responsibility for raising immunization levels among preschoolers in medically underserved communities . Communities with federally designated health professional shortage areas were invited to submit proposals, and 4 were selected: Detroit, MI, New York, NY, San Diego, CA, and rural Colorado . OBJECTIVES: To measure immunization coverage among preschool children in the 4 selected medically underserved areas and determine predictors of coverage levels . DESIGN AND SETTING: Surveys in the 4 areas were based on stratified cluster probability sample designs in which clusters of dwelling units were selected and all households in selected clusters were screened for the presence of children aged 12 to 35 months . Immunization histories were obtained from parents and providers for these children . For each age-eligible child, the information collected on utilization of immunization health services included a listing of all clinics or offices ever used for the child's well-child care and/or for obtaining immunizations . Information was also collected on whether the child currently had health insurance (public and/or private) and whether the child had a medical home . A child was classified as having a medical home if the survey respondent reported a source of well care that was the same as the source of sick care and that this place was not an emergency department . PARTICIPANTS: Children 12 to 35 months of age in Detroit, New York, San Diego, and rural Colorado . OUTCOME MEASURE: Community-wide up-to-date (UTD) immunization coverage levels at 19 to 35 months of age, defined as receipt of 4 doses of diphtheria and tetanus toxoids and pertussis vaccine, 3 doses of poliovirus vaccine, 1 dose of measles, mumps, and rubella vaccine, 3 doses of Haemophilus influenzae type B vaccine, and 3 doses of hepatitis B vaccine (the 4:3:1:3:3 series) . ANALYSIS: We examined the association between coverage level and independent variables and performed chi2 and t tests to determine whether differences observed within and between groups and sites were significant . RESULTS: The overall response rate for eligible children ranged from 79.4% to 88.1% . Coverage levels for most individual vaccines were >90% in all sites except Detroit . Coverage for the 4:3:1:3:3 series was significantly higher for children in New York (84%) and San Diego (86%) than for children in Detroit (66%) and rural Colorado (75%) . Demographic risk factors related to UTD immunization status varied by site . Although differences in coverage levels by ethnicity varied by site, differences were not significant . In Colorado and New York, coverage was slightly lower among Hispanic than white children (71% vs 76% and 83% vs 91%, respectively) . In San Diego, coverage was lower among whites, compared with Hispanics (76% vs 85%) . Coverage was also lower for African American than white children only in New York (75% vs 91%) . However, in San Diego and Colorado, children receiving their vaccinations from private providers had lower coverage levels than children receiving their vaccinations from other providers (78% vs 91% and 71% vs 57%, respectively) . Ictively) . In all 4 sites, children for whom respondents reported having an immunization card at the time of the interview were more likely to have higher series coverage levels than children for whom a parent-held card was not available . Also, children who were UTD at 3 months of age had significantly higher vaccination-series coverage levels than children who were not UTD at 3 months of age . In addition, the vaccination coverage was lower for children in Detroit whose parents reported problems accessing the health care system because lack of transportation (46%), compared with those who did not report such problems (65%); however, this difference did not reach significance (chi2 = 6.0) . In Colorado, the small proportion of children in families without a phone had a lower vaccination coverage level (58%) than those in households with a phone (75%) (chi2 = 6.3) . In all sites, children who were UTD at 3 months of age and had a parent-held vaccination card were more likely to be UTD at 19 to 35 months of age . CONCLUSIONS: Preschoolers in these medically underserved areas were not at uniform risk for underimmunization . Because they were designated as health professional shortage areas, the 4 sites in this study were expected to have low immunization-coverage rates . However, this was not the case . In fact, coverage in 3 of the 4 areas was quite high compared with US national figures (73%); only Detroit had a much lower UTD rate (66%) . Efforts are needed to improve methods to identify areas with low immunization coverage so that resources can be directed to places where interventions are needed . Our results reveal that an area's need for childhood immunization interventions is not well predicted by a low number of providers per capita . Other criteria must be developed to predict areas or populations with low immunization coverage . Understanding more about the characteristics of children/provider pairs for children who are UTD at 3 months and more about the role of parental hand-held cards, along with finding strategies to improve immunization delivery by providers in Vaccines for Children Program facilities, suggest potentially productive avenues for increasing and sustaining high coverage levels.

Drugs, 2004, 64(8), 805 - 19
Acute sinusitis: guide to selection of antibacterial therapy; Lindbaek M; Primary care doctors should be cautious in the diagnosis and treatment of sinusitis as acute bacterial sinusitis is currently over-diagnosed and over-treated in primary care practice . The clinical diagnosis of acute bacterial sinusitis is difficult in primary care practice; however, a history of purulent rhinorrhoea, purulent secretions in the nasal cavity on examination, tooth pain, worsening of symptoms following initial improvement, lack of effect of decongestants and an elevated erythrocyte sedimentation rate are supportive evidence of bacterial infection . Patients with symptoms for <7 days are not as likely to have bacterial infection . Acute sinusitis is over-treated in primary care practice for several reasons . Firstly, most cases of acute sinusitis are caused by viral infections and resolve without antibacterial treatment . Secondly, in clinical trials of antibacterial treatment, only about one-half of patients diagnosed with acute bacterial sinusitis by experienced primary care physicians have bacterial infection . Thirdly, antibacterial treatment of acute sinusitis is indicated only in patients with severe symptoms of sinusitis or in patients with moderate symptoms of >7 days duration . Symptomatic treatment is sufficient in patients with mild symptoms . Three recent meta-analyses have concluded that newer and broad-spectrum antibacterials are not significantly more effective than narrow-spectrum agents, such as amoxicillin or phenoxymethylpenicillin (penicillin V) . However, because of the rapid increase in antibacterial resistance of Streptococcus pneumoniae and Haemophilus influenzae, treatment must take into account current recommendations for treating infections caused by these organisms . Fourthly, sinus imaging studies are not recommended in routine diagnosis but may be helpful in selected cases . Finally, other than pain medication, there is little evidence that use of adjunctive treatments, such as decongestants, is effective in symptom relief . However, a recent study in patients with recurrent sinusitis demonstrated that patients who received fluticasone propionate in addition to antibacterials had a higher rate of clinical success than did patients receiving placebo and antibacterials.

Przegl Lek, 2003, 60(11), 699 - 703
{Seroconversion after vaccination against pertussis, Haemophilus influenzae type b and poliomyelitis in preterm infants}; Sikora JP et al.; OBJECTIVE: This study was aimed to determine the levels of antibodies against Bordetella pertussis, Haemophilus influenzae type b (Hib) and Poliovirus type 1 in preterm children after immunization with acellular pertussis (Infanrix-DTPa) and Hib conjugate (Hiberix) vaccines as well as inactivated vaccine against poliomyelitis (Imovax Polio) . MATERIAL AND METHODS: 32 children were given 3 doses of Infanrix-DTPa and Hiberix vaccines as well as Imovax Polio vaccine . Samples of blood were taken from children 4 weeks after application the 3-rd dose of vaccines and compared with the initial levels of antibodies (seroconversion was evaluated in 32 children immunized with both infanrix-DTPa and Hiberix and in 20 ones immunized with Imovax Polio) . We evaluated the postvaccinal immunity against Bordetella pertussis and Hib by means of Immunoenzymatic methods (ELISA), with commercially available test kits (Bordetella pertussis ELISA KIT, Genzyme Virotech GmbH, Russelsheim, Germany and BINDAZYME Human Anti Haemophilus Influenzae Enzyme Immunoassay Kit, The Binding Site, Birmingham, U.K.); antibodies against Poliovirus type 1 were assessed using neutralisation method . RESULTS: All the children (100%) demonstrated protective levels of antibodies after Immunization with 3-rd doses of Infanrix-DTPa and Hiberix . Only one child (5%) didn't respond when Imovax Polio was applied . The observed increase of postvaccinal antibodies titers as to all applied antigens was statistically significant (p < 0.05) . No significant correlations between birth weight, gestational age and the achieved levels of postvaccinal antibodies were noticed . CONCLUSIONS: Our results prove that nearly all the preterm infants achieve the protective levels of antibodies against pertussis, Hib and poliomyelitis . Hiberix due to its high immunogenicity should be applied in the obligatory vaccination schedules, especially in preterm infants.

Infection, 2004 Apr, 32(2), 112 - 5
Meningitis due to Haemophilus influenzae type f in an 8-year-old girl with congenital humoral immunodeficiency; Fickweiler K et al.; We report on an 8-year-old Haemophilus influenzae type b-vaccinated girl with meningitis due to H . influenzae type f . The girl had an oro-facio-digital syndrome and a hypogammaglobulinemia, obviously predisposing the patient to invasive infections . Treatment with cefotaxime was successful.Incidence, predisposing factors and the influence of H . influenzae type b immunization on invasive infections with non-type b H . influenzae strains in children are discussed.

J Antimicrob Chemother, 2004 May, 53(5), 793 - 6 Epub 2004 Mar 31.
Comparative in vitro activity of telithromycin against macrolide-resistant and -susceptible Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae; Walsh F et al.; OBJECTIVES: The first objective was to investigate the in vitro activity of telithromycin against respiratory tract pathogens in comparison with other antimicrobial agents . The second objective was to identify the influence of the erm(B) and mef(A) genes on the susceptibility of Streptococcus pneumoniae to telithromycin . METHODS: The in vitro activity of telithromycin against S . pneumoniae, Moraxella catarrhalis and Haemophilus influenzae, isolated from the UK and 40 macrolide-resistant S . pneumoniae from four different countries was compared with a variety of antimicrobial agents . The 140 isolates were examined for the presence of the erm(B) and mef(A) genes . The impact of 5% CO(2) on susceptibility testing was also investigated . RESULTS: Telithromycin showed greatest activity against S . pneumoniae, but also had good activity against M . catarrhalis and H . influenzae, which was independent of their resistance profiles to other antibiotics . The MIC(90) of telithromycin for S . pneumoniae was 0.12 mg/L, which was 64-fold lower than the lowest macrolide MIC; 21% of the S . pneumoniae were macrolide resistant . Thirty-eight per cent of the macrolide-resistant strains were erm(B)-positive and 62% were mef(A)-positive, but no strain contained both genes . The activity of telithromycin was similar to that of azithromycin against both M . catarrhalis and H . influenzae, Erythromycin was slightly less active: 1% and 8% of M . catarrhalis and H . influenzae, respectively, were resistant to erythromycin, but none were resistant to telithromycin . Five per cent of the S . pneumoniae strains and 4% of the H . influenzae strains changed from telithromycin susceptible to non-susceptible entirely because of the incubation conditions . The MIC(50)s and MIC(90)s of S . pneumoniae, M . catarrhalis and H . influenzae increased by one dilution when incubated in CO(2) . CONCLUSIONS: Telithromycin has shown high in vitro activity against S . pneumoniae, including those strains that are macrolide susceptible and resistant as well as M . catarrhalis and H . influenzae . This study has also demonstrated that there is no cross-resistance between erythromycin and telithromycin . The impact of 5% CO(2) on susceptibility testing should be investigated further before providing definite guidelines on telithromycin susceptibility testing.

Enferm Infecc Microbiol Clin, 2004 Apr, 22(4), 230 - 7
{Bacterial resistance and pharmacodynamics as the basis for prescribing antibiotics in respiratory infections}; Aguado-Garcia JM et al.; The choice of antibiotic therapy in respiratory tract infections is usually empirical . However, this choice is complicated by the increasing prevalence of resistant strains among the major bacterial pathogens involved in these infections, particularly Streptococcus pneumoniae . The aim of antimicrobial therapy in respiratory tract infections should be bacterial eradication, which is necessary to maximize clinical cure and minimize the development and spread of resistance . An increase in antimicrobial resistance reduces the probability of achieving eradication and increases the probability of clinical failure . Recent reports have demonstrated the clinical relevance of respiratory bacterial resistance to macrolides and some fluoroquinolones and betalactams . Unlike macrolide and fluoroquinolone resistance, penicillin resistance in Streptococcus pneumoniae can be overcome by increasing the dose, and hence increasing the time during which serum concentrations are above the MIC . Pharmacokinetic/pharmacodynamic (PK/PD) parameters can be used to establish breakpoints predictive of bacterial eradication . From the viewpoint of PK/PD, in Spain only high-doses of amoxicillin/clavulanic acid (875/125 mg tid and 2000/125 mg bid) and levofloxacin, among the oral antibiotics considered, achieve optimal coverage against S . pneumoniae and Haemophilus influenzae.

Enferm Infecc Microbiol Clin, 2004 Apr, 22(4), 220 - 6
{Observational study investigating the use of levofloxacin in ICU patients}; Alvarez-Lerma F et al.; Introduction . There is little information on the use of levofloxacin, a new quinolone, in ICU patients . Objective . To investigate the criteria for the use of levofloxacin (indications, forms of prescription, doses, and routes of administration) and to study tolerance in patients admitted to the ICU . Method . Prospective, observational study performed from October 2000 to November 2001 in 35 ICUs and including the first 15 patients receiving levofloxacin as monotherapy or combined treatment . Descriptive data are expressed as mean and percentage . Statistical significance was set at P <.05 . Results . A total of 543 indications for treatment with levofloxacin were analyzed . The patients were 70.7% men, with a mean (SD) age of 60.2 (16.7) years, mean APACHE II score of 18.9 (7.9), and a medical underlying disease in 79.2% of cases . The ICU mortality rate was 24.1% . A total of 60% of patients required mechanical ventilation and 44.3% needed inotropic drug treatment . Levofloxacin was predominantly prescribed for treating community-acquired infections (67.8%), mainly in the respiratory tract (88.1%) . An etiological diagnosis was established in only 55.6% of cases . The most common pathogens were Streptococcus pneumoniae (12.7%), Haemophilus influenzae (9.1%), Escherichia coli (7.4%), methicillin-sensitive Staphylococcus aureus (7.2%), Pseudomonas aeruginosa (4.9%), and Legionella pneumophila (4.7%) . In 87.1% of indications, levofloxacin was prescribed as empirical treatment . Susceptibility of the isolated pathogens to this antibiotic was confirmed in 32.2% of cases . The initial dose was 500 mg/24 h in 48.5% of indications and 500 mg/12 h in 48.3% . Combined treatment was given in 49.7% of cases . In 32.2% of cases, parenteral administration of levofloxacin was changed to oral route . Adverse events probably or possibly associated with levofloxacin occurred in only 12.5% of patients and mainly included increased ALT/ALS levels (4.4%), diarrhea (2.3%), and heart rhythm alterations (2.1%) . Conclusions . This study describes the profile of critically ill patients receiving levofloxacin and the different forms of its use in the ICU.

Otolaryngol Head Neck Surg, 2004 Mar, 130(3), 339 - 43
Vaccine-preventable pediatric postmeningitic sensorineural hearing loss in southern India; Cherukupally SR et al.; OBJECTIVE: The study goal was to assess vaccine-preventable pediatric postmeningitic sensorineural hearing loss in southern India . STUDY DESIGN AND SETTING: We conducted a prospective pilot study from January through March 2001 in a tertiary pediatric hospital in southern India . RESULTS: Sixty-five patients were studied . Thirty-five (54%) patients had positive cerebrospinal fluid cultures, with the most common organisms being Streptococcus pneumoniae and Haemophilus influenzae . An additional 10 (15%) patients were diagnosed with tuberculous meningitis . Of 28 patients who could comply with audiometric evaluations, 8 (28%) had sensorineural hearing loss . CONCLUSIONS: The etiologic organisms of bacterial meningitis in this study population are similar to those organisms in the developed world, with the minority exception of tuberculous meningitis . Further, a similar prevalence of postmeningitic sensorineural hearing loss occurred . SIGNIFICANCE: An effective vaccination program against S . pneumoniae and H . influenzae type b should reduce the prevalence of sensorineural hearing loss due to bacterial meningitis in developing countries with similar bacterial profiles.

Sex Transm Infect, 2004 Apr, 80(2), 124 - 9
Review of STI and HIV epidemiological data from 1990 to 2001 in urban Burkina Faso: implications for STI and HIV control; Nagot N et al.; OBJECTIVES: To better understand the sexually transmitted infection (STI)/HIV dynamics in an urban west African setting in order to adapt STI/HIV control efforts accordingly . METHODS: Review of STI and HIV epidemiological studies performed over the past decade in Bobo-Dioulasso, the second city of Burkina Faso . Trends in STI prevalence among commercial sex workers and the general population were assessed over time through studies that used the same recruitment and laboratory diagnostic procedures . Variations in aetiologies of vaginal discharge, urethral discharge, and genital ulcers were also evaluated among patients consulting for genital infection complaints . Antenatal clinic based surveys provided data to assess HIV trend among the general population . RESULTS: We observed an important decline of classic bacterial STI such as syphilis, Neisseria gonorrhoea, Chlamydia trachomatis, and Haemophilus ducrey infections in all study groups . Trichomoniasis also declined but to a lesser extent . HIV infection followed the same trend at the same time, with a significant decline in the 15-19 year age group of pregnant women, suggesting a possible decrease of HIV incidence . Although no evidence of a causal relation can be drawn from this review, adoption of safer sex behaviour, introduction of the syndromic management (SM) approach, or higher antibiotic use may have contributed to these changes . CONCLUSIONS: Classic bacterial STI declined over the past decade in parallel with a stabilisation of HIV infection . Variations in syndromes aetiology and sexual behaviours should be monitored as part of STI surveillance in order to improve STI syndromic management algorithms and to adapt HIV/STI prevention efforts.

J Comp Pathol, 2004 May, 130(4), 294 - 8
In-situ hybridization for the detection of Haemophilus parasuis in naturally infected pigs; Jung K et al.; Detection of Haemophilus parasuis in naturally infected pigs was studied by in-situ hybridization with a non-radioactive digoxigenin-labelled DNA probe . Twenty pigs were selected on the basis of bacterial isolation and histopathological lesions . An 821 base pair DNA probe from the 16S small subunit ribosomal RNA (rRNA) was generated by the polymerase chain reaction . Hybridization signals were detected in formalin-fixed, paraffin-wax-embedded tissues (lung, heart, spleen and liver) . Identification of the cell types containing H . parasuis was occasionally difficult, but examination of adjacent sections stained with haematoxylin and eosin confirmed that positive cells resembled either macrophages (large oval nuclei and abundant cytoplasm) or neutrophils (bilobed nuclei) . In-situ hybridization would appear to be a valuable tool for the diagnosis of H . parasuis infection.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1416 - 8
Identification of the Haemophilus influenzae tolC gene by susceptibility profiles of insertionally inactivated efflux pump mutants; Trepod CM et al.; Isogenic strains containing insertional disruptions of 10 Haemophilus influenzae Rd genes were investigated for their effects on the susceptibility of the organism to various classes of antimicrobial compounds . MIC results show that HI1462, which encodes an Escherichia coli TolC homolog, is the third component of the H . influenzae AcrAB pump.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1229 - 34
In vitro activities of piperacillin against beta-lactamase-negative ampicillin-resistant Haemophilus influenzae; Morikawa Y et al.; The in vitro activities of piperacillin (PIP) against beta-lactamase-negative ampicillin (AMP)-resistant (BLNAR) Haemophilus influenzae were compared with those of cefotaxime (CTX) and ceftriaxone (CRO), and the potency of PIP as therapy for meningitis caused by BLNAR is also discussed . PIP showed good activity (MIC at which 90% of strains are inhibited, 0.25 micro g/ml) against 69 BLNAR strains, and its activity was comparable to that of CRO and superior to that of CTX . No significant correlation was observed between the MICs of PIP and CTX or CRO or AMP, whereas a high correlation was observed between the MICs of CTX and CRO . In the killing study, PIP showed potent bactericidal activity compared with those of CTX and CRO . By microscopic examination, PIP caused the formation of a spindle and short filamentous cells with bulges and induced cell lysis in BLNAR strains, while treatment with CTX and CRO resulted in the formation of large, spherical cells without any obvious lysis . The affinity of Bocillin FL, a fluorescent penicillin used for determination of the 50% inhibitory concentration (IC(50)s) for penicillin-binding proteins (PBPs), to PBPs 3a and 3b of BLNAR strains was drastically decreased compared with that to an AMP-susceptible strain (ATCC 33391) . In the case of the BLNAR strains, the IC(50)s for PBPs 1a, 1b, and 2 were similar to those for the PBPs of ATCC 33391 . Since the affinity of binding to PBPs 3a and 3b of the BLNAR strains decreased drastically, the second targets among the PBPs were PBP 2 for PIP, PBP1 (1a and 1b) for CTX and CRO . In conclusion, PIP showed excellent activities against BLNAR strains in a manner different from those of cephem antibiotics, suggesting that it could be a candidate therapeutic agent for the treatment of meningitis caused by BLNAR strains.

J Mol Microbiol Biotechnol, 2003, 6(2), 109 - 26
A global approach to identify novel broad-spectrum antibacterial targets among proteins of unknown function; Zalacain M et al.; Attempted allelic replacement of 144 Streptococcus pneumoniae open reading frames of previously uncharacterized function led to the identification of 36 genes essential for growth under laboratory conditions . Of these, 14 genes (obg, spoIIIJ2, trmU, yacA, yacM, ydiC, ydiE, yjbN, yneS, yphC, ysxC, ytaG, yloI and yxeH4) were also essential in Staphylococcus aureus and Haemophilus influenzae or Escherichia coli, 2 genes (yrrK and ydiB) were only essential in H . influenzae as well as S . pneumoniae and 8 genes were necessary for growth of S.pneumoniae and S . aureus and did not have a homolog in H . influenzae(murD2, ykqC, ylqF, yqeH, ytgP, yybQ) or were not essential in that organism (yqeL, yhcT) . The proteins encoded by these genes could represent good targets for novel antibiotics covering different therapeutic profiles . The putative functions of some of these essential proteins, inferred by bioinformatic analysis, are presented . Four mutants, with deletions of loci not essential for in vitro growth, were found to be severely attenuated in a murine respiratory tract infection model, suggesting that not all targets for antibacterial therapeutics are revealed by simple in vitro essentiality testing . The results of our experiments together with those collated from previously reported studies including Bacillus subtilis, E . coli and Mycoplasma sp . demonstrate that gene conservation amongst bacteria does not necessarily indicate that essentiality in one organism can be extrapolated to others . Moreover, this study demonstrates that different experimental procedures can produce apparently contradictory results .

Infect Immun, 2004 Apr, 72(4), 1874 - 84
The LspB protein is involved in the secretion of the LspA1 and LspA2 proteins by Haemophilus ducreyi; Ward CK et al.; The LspA1 and LspA2 proteins of Haemophilus ducreyi 35000 are two very large macromolecules that can be detected in concentrated culture supernatant fluid . Both of these proteins exhibit homology with the N-terminal region of the Bordetella pertussis filamentous hemagglutinin (FHA), which is involved in secretion of the latter macromolecule . The lspA2 open reading frame is flanked upstream by a gene, lspB, that encodes a predicted protein with homology to the B . pertussis FhaC outer membrane protein that is involved in secretion of FHA across the outer membrane . The H . ducreyi lspB gene encodes a protein with a predicted molecular mass of 66,573 Da . Reverse transcription-PCR analysis suggested that the lspB gene was transcribed together with the lspA2 gene on a single mRNA transcript . Polyclonal H . ducreyi LspB antiserum reacted with a 64-kDa antigen present in the Sarkosyl-insoluble cell envelope fraction of H . ducreyi 35000, which indicated that the LspB protein is likely an outer membrane protein . Concentrated culture supernatant fluids from H . ducreyi lspB and lspA1 lspB mutants did not contain detectable LspA1 and detectable LspA2, respectively . However, complementation of the lspB mutant with the wild-type lspB gene on a plasmid restored LspB protein expression and resulted in release of detectable amounts of the LspA1 protein into culture supernatant fluid . When evaluated in the temperature-dependent rabbit model of infection, the lspB mutant was attenuated in the ability to cause lesions and was never recovered in a viable form from lesions . These results indicated that the H . ducreyi LspB protein is involved in secretion of the LspA1 and LspA2 proteins across the outer membrane.

Mol Immunol, 2004 Feb, 40(14-15), 1103 - 8
Innate and adaptive immune requirements for induction of autoimmune demyelinating disease by molecular mimicry; Olson JK et al.; Molecular mimicry is the main postulated mechanism by which infectious agents induce autoimmune disease . A number of animal models have been utilized to establish a link between molecular mimicry and autoimmunity . However, a model of infectious disease whereby a natural pathogen expressing a known mimic epitope can induce autoimmunity to a known self-antigen leading to clinical autoimmune disease is still lacking . We have engineered a recombinant Theiler's murine encephalomyelitis virus (TMEV) to express an encephalitogenic myelin proteolipid protein PLP139-151 epitope (PLP-TMEV) and a PLP139-151 mimic peptide naturally expressed by Haemophilus influenzae (HI-TMEV) . Infection of mice with either PLP-TMEV or HI-TMEV induces early-onset disease that is associated with the activation of cross-reactive PLP139-151-specific immunopathologic CD4+ Th1 cells . Based on results from this model, we hypothesize, due to the considerable degeneracy in the T cell repertoire, that induction of full-blown autoimmune disease via molecular mimicry is a tightly regulated process requiring multiple factors related to the pathogen expressing the potential mimic epitope . In this review, we will discuss how various factors related to the infectious environment control whether or not autoimmune disease is initiated . Contributing factors include the nature of the innate immune response to the pathogen which determines the immunopathologic potential of the induced cross-reactive T cells, the capacity of the mimic epitope to be processed and presented from its natural flanking sequences in the pathogen-encoded protein, the site(s) of the primary infection in the host and the ability of the pathogen to persist, and the potential requirement for multiple infections with the same or different pathogens.

Cytokine, 2004 Mar 21, 25(6), 254 - 9
Effects of amoxicillin on cytokine and osteocalcin expression in bone tissue during experimental acute otitis media; Melhus A et al.; There are indications that bone regulatory and immune systems are closely related . Of special interest in this context is the acute otitis media (AOM), which mainly affects immunologically immature patients and, when complicated, involves bone tissue . To explore host responses in bone tissue during Haemophilus influenzae-induced AOM modified by amoxicillin, a rat model and PCR techniques were used . The treatment eradicated the bacteria and induced changes in the expression profile of osteocalcin, a bone formation marker . The maximum levels of osteocalcin transcripts in the treatment group were delayed by about a week . The mRNA levels never reached the same high levels as in the untreated animal group, but the downregulation was slow and entailed higher osteocalcin mRNA levels for a longer time period in the treated animals . The expression of IL-6 and TNF-alpha, two cytokines associated with bone resorption, remained unaffected by the amoxicillin treatment, whereas the downregulation of IL-10, with suppressive effects on bone resorption, was slower than that during the natural course . By comparing the host responses on the molecular level in different tissues during treated and untreated AOM, new approaches to how to minimize the risks of severe AOM complications may evolve.

J Am Acad Dermatol, 2004 Apr, 50(4), 495 - 528; quiz 529-32
Vaccines and immunotherapies for the prevention of infectious diseases having cutaneous manifestations; Wu JJ et al.; Although the development of antimicrobial drugs has advanced rapidly in the past several years, such agents act against only certain groups of microbes and are associated with increasing rates of resistance . These limitations of treatment force physicians to continue to rely on prevention, which is more effective and cost-effective than therapy . From the use of the smallpox vaccine by Jenner in the 1700s to the current concerns about biologic warfare, the technology for vaccine development has seen numerous advances . The currently available vaccines for viral illnesses include Dryvax for smallpox; the combination measles, mumps, and rubella vaccine; inactivated vaccine for hepatitis A; plasma-derived vaccine for hepatitis B; and the live attenuated Oka strain vaccine for varicella zoster . Vaccines available against bacterial illnesses include those for anthrax, Haemophilus influenzae, and Neisseria meningitidis . Currently in development for both prophylactic and therapeutic purposes are vaccines for HIV, herpes simplex virus, and human papillomavirus . Other vaccines being investigated for prevention are those for cytomegalovirus, respiratory syncytial virus, parainfluenza virus, hepatitis C, and dengue fever, among many others . Fungal and protozoan diseases are also subjects of vaccine research . Among immunoglobulins approved for prophylactic and therapeutic use are those against cytomegalovirus, hepatitis A and B, measles, rabies, and tetanus . With this progress, it is hoped that effective vaccines soon will be developed for many more infectious diseases with cutaneous manifestations.

J Anim Sci, 2004 Mar, 82(3), 837 - 44
Effects of dietary energy and starch concentrations for newly received feedlot calves: I . Growth performance and health; Berry BA et al.; Crossbred calves (n = 572; initial BW = 186 +/- 27 kg) purchased from northern Texas, Arkansas, and southeast Oklahoma auction markets were delivered to the Willard Sparks Beef Research Center, Stillwater, OK, and used to study the effects of dietary energy and starch concentrations on performance and health of newly received feedlot calves during a 42-d receiving period . On arrival, calves were assigned randomly to one of two dietary energy levels (0.85 or 1.07 Mcal NEg/kg DM) and one of two dietary starch levels (34 or 48% of ME from starch) in a 2 x 2 factorial arrangement of treatments . Cattle were weighed and serum samples were collected on d 0, 7, 14, 28, and 42 . Individual animal records of morbidity were kept for all cases of respiratory and other disease . Nasal swabs were collected from each morbid animal and cultured for upper-respiratory pathogens . There were no energy x starch level interactions for performance or health response variables . Daily gain (1.14 kg/d) and gain efficiency (ADG:DMI = 0.179) were not affected by increasing dietary energy or starch concentrations . Calves fed low-energy diets consumed (P < 0.05) more DM . No difference (P = 0.54) was detected in morbidity for calves fed high-energy (62.4% calves treated) compared with low-energy (65.8% calves treated) diets; however, calves fed the high-starch diets had numerically (P = 0.11) greater morbidity than calves fed low-starch diets (68.8 vs . 59.4% calves treated, respectively) . There were no energy or starch effects on Mannheimia haemolytica or Pasteurella multocida antibody titers; however, day effects (P < 0.02) occurred . On d 7, 14, and 28, calves had antibody titers for P . multocida that were greater (P < 0.05) than titers on d 0 . In addition, calves had greater antibody titers to M . haemolytica on d 7 and 14 than on d 0 . Nasal swabs revealed that calves fed the high-energy diets tended (P = 0.06) to have a lower percentage of morbid calves with P . multocida during the first antimicrobial treatment and a lower percentage of Haemophilus somnus isolates during the first (P = 0.01) and second (P = 0.06) antimicrobial treatments than calves fed the low-energy diets . Although animal performance was not influenced, the present data suggest that feeding the high-energy diet decreased the percentage of P . multocida and H . somnus pathogens in calves that received one or more antimicrobial treatments.

Nippon Jibiinkoka Gakkai Kaiho, 2004 Feb, 107(2), 156 - 68
{Antibiotic selection for the treatment of pediatric upper respiratory tract infections and the detection rate of drug-resistant strains}; Tomiyama M; In recent years, Streptococcus pneumoniae and Haemophilus influenzae have acquired resistance to some drugs; as such, a therapeutic strategy in which penicillins (PCs) are primarily used is currently recommended for the treatment of pediatric upper respiratory tract infections . In 2000, we changed the primary drug used for such infections in patients visiting our hospital from cephems (CEPs) to PCs . We examined the effect of this change on the detection rates of drug-resistant S . pneumoniae (DRSP) and ampicillin (ABPC)-resistant H . influenzae . 1 . The subjects consisted of patients who had visited our hospital and received bacteriological examinations during 1999, 2001 or 2003 . At the time of the examination, we questioned each patient regarding the medical institutions where they had received oral antibiotic therapy during the past year, how many antibiotic therapies they had received, and which antibiotics they had received . The number of patients who had only received antibiotics from our hospital was 37 out of 52 (71%) patients in 1999, 80 out of 110 (73%) patients in 2001, and 50 out of 71 (70%) patients in 2003 . The number of therapies in which only PCs were used was significantly higher in 2001 and 2003 than in 1999 . The number of therapies in which only CEPs were used was lower in 2001 and 2003 than in 1999 . We believe that this trend reflects our therapeutic strategy . Furthermore, the antibiotic therapies that patients received at other hospitals were thought to have only a small influence on the detection rates of DRSP and ABPC-resistant H . influenzae . 2 . The detection rate for DRSP was 30% in 1995 and 60% in 1999 . However, the detection rate for DRSP was 37% in 2001 and 39% in 2003, representing a significant decrease from the detection rate in 1999 . The detection rate for ABPC-resistant H . influenzae was 26% in 1995 and increased slightly to 36% in 1999 . However, the detection rate for ABPC-resistant H . influenzae increased significantly from 36% in 1999 to 59% in 2001 and to 71% in 2003 . The reason for this trend is that PCs are considered to be less effective than CEPs against H . influenzae, even though they are more effective than CEPs against S . pneumoniae . 3 . We received the patients at our hospital who required hospitalization for the treatment of bacterial infections . In 1999, DRSP was detected in 2 of the 11 (18%) patients who required hospitalization, but no ABPC-resistant H . influenzae strains were detected . ABPC-resistant H . influenzae was detected in 3 of the 12 (25%) patients hospitalized in 2001 and in 4 of the 10 (40%) patients hospitalized in 2003 . Thus, the detection rate for ABPC-resistant H . influenzae appears to be increasing . 4 . Our findings suggest that the number of DRSP isolates may increase when treatment policies focused on CEPs are employed, while the number of ABPC-resistant H . influenzae isolates may increase when treatment policies are focused on PCs . 5 . At present, guidelines focusing on the use of PCs for the treatment of pediatric upper respiratory tract infection are recommended, but standardizing the choice of antibiotics may cause the number of resistant bacteria to increase . When such guidelines are employed at medical institutions, attention should be constantly paid to trends in drug susceptibility . If an increase in one type of resistant bacteria is observed, flexible measures should be taken, without strict adherence to the guideline.

J Infect Dis, 2004 Apr 1, 189(7), 1304 - 12 Epub 2004 Mar 17.
Intranasal vaccination with recombinant P6 protein and adamantylamide dipeptide as mucosal adjuvant confers efficient protection against otitis media and lung infection by nontypeable Haemophilus influenzae; Bertot GM et al.; Nontypeable Haemophilus influenzae (NTHi) is a leading etiologic agent of otitis media in children and recurrent respiratory infections in patients with chronic obstructive pulmonary disease . The highly conserved outer membrane protein P6 constitutes a promising vaccine candidate antigen . However, the small amount of P6 produced by this fastidious microorganism renders large-scale production difficult . Controversial data also exist concerning the suitability of recombinant P6 (rP6) as a vaccine antigen . Therefore, we performed a comparative evaluation of the immunogenicity and efficacy of native P6 and rP6 in mice intranasally vaccinated with adamantylamide dipeptide (AdDP) as an adjuvant . High titers of P6-specific serum antibodies were elicited in mice vaccinated with either native P6 or rP6, which cross-recognized both antigens . However, rP6 stimulated stronger mucosal responses . Mice vaccinated with rP6 were protected against both pulmonary and middle-ear infections (P<.01) . This demonstrates that rP6 plus AdDP constitutes a promising vaccine formulation against the most relevant forms of disease caused by NTHi.

Brain Dev, 2004 Apr, 26(3), 168 - 75
Bacterial meningitis in infants: the epidemiology, clinical features, and prognostic factors; Chang CJ et al.; This 16-year (1986-2001) retrospective study enrolled 80 infantile patients (aged, 30-365 days old) with culture-proven bacterial meningitis . The most prevalent pathogens were Salmonellaspecies, Streptococcus (S.) agalactiae, Escherichia (E.) coli, and Haemophilus (H.) influenzae, accounting for about 59% of the episodes . Meningitis caused by Salmonella species, E . coli and H . influenzae occurs more often in the older infants, while that caused by S . agalactiae occurs more often in young infants . Our study revealed a decrease in the proportion of Salmonella meningitis from 27% in the first 8 years to 9% in the second 8 years with E . coli replacing Salmonella species as the leading pathogen of this disease during the second period . Overall mortality rate for both periods of time was 11% . However, if we take those with undesirable poor outcomes into account, 43% of patients could be considered treatment failures . The study also reveals a high prevalence of neurological complications when this disease is caused by H . influenzae, S . pneumoniae, and Salmonella species . Stepwise logistic regression analysis revealed that only initial changing levels of consciousness (P = 0.006) were independently associated with treatment failure . The most frequent neurological complications associated with this disease included subdural empyema, hydrocephalus, cerebral infarctions, and seizures . Because therapeutic regimens may require attention to the eradication of bacterial pathogen but also the neurological complications, early diagnosis and choice of appropriate antibiotics are essential to increasing the possibility of survival.

Ann Fr Anesth Reanim, 2004 Mar, 23(2), 138 - 41
{Uncommon cause of respiratory distress in the post-anaesthesia care unit}; Lalot M et al.; We report a case of respiratory distress in the post anesthesia care unit following general anaesthesia for a dilatation and curettage related to miscarriage in a 32-year-old woman . The preoperative physical examination showed no abnormalities except for the presence of dry cough during the preceding two or three days . A few minutes after her arrival in the PACU, the patient developed hyperthermia till 40.6 degrees C, cough, polypnea and oxygen desaturation (SpO(2): 82% on FiO(2): 40%) . A thoraco-abdominal angioscanner showed pulmonary basal condensations and a thrombosis of the right ovarian vein . The patient had to be transferred to the intensive care unit where she remained intubated and ventilated during 13 days because of a Haemophilus influenzae pneumonia.

EMBO J, 2004 Mar 24, 23(6), 1245 - 56 Epub 2004 Mar 18.
Structural basis for host recognition by the Haemophilus influenzae Hia autotransporter; Yeo HJ et al.; Haemophilus influenzae is an important human pathogen that initiates infection by colonizing the upper respiratory tract . The H . influenzae Hia autotransporter is an adhesive protein that promotes adherence to respiratory epithelial cells . Hia adhesive activity resides in two homologous binding domains, called HiaBD1 and HiaBD2 . These domains interact with the same host cell receptor, but bind with different affinities . In this report, we describe the crystal structure of the high-affinity HiaBD1 binding domain, which has a novel trimeric architecture with three-fold symmetry and a mushroom shape . The subunit constituents of the trimer are extensively intertwined . The receptor-binding pocket is formed by an acidic patch that is present on all three faces of the trimer, providing potential for a multivalent interaction with the host cell surface, analogous to observations with the trimeric tumor necrosis factor superfamily of proteins . Hia is a novel example of a bacterial trimeric adhesin and may be the prototype member of a large family of bacterial virulence proteins with a similar architecture.

Clin Pediatr (Phila), 2004 Mar, 43(2), 135 - 51
Optimizing antibacterial therapy for community-acquired respiratory tract infections in children in an era of bacterial resistance; Low DE et al.; The spread of antibacterial resistance in bacteria that commonly cause childhood community-acquired respiratory tract infections (RTIs), such as acute otitis media, community-acquired pneumonia, and acute pharyngitis, is a major healthcare problem . One of the foremost concerns is the rapid increase in penicillin, macrolide, and multidrug resistance in Streptococcus pneumoniae . There is also a rising prevalence of macrolide resistance in Streptococcus pyogenes in pockets of the United States, and beta-lactamase production in Haemophilus influenzae is widespread . Although data are limited, some evidence suggests that resistance to antibacterials can impair bacteriologic and clinical outcomes in childhood RTIs . Optimizing antibacterial use is important both in the care of individual patients and within strategies to address the wider problem of antibacterial resistance . This involves encouraging judicious antibacterial use (i.e., reducing overuse for viral infection and prophylaxis), and preventing misuse through the wrong choice, dosage, and duration of therapy . Given that initial therapy is usually empiric, antibacterials used to treat community-acquired RTIs in children should ideally have the following properties: an optimal targeted spectrum of activity; high clinical and bacteriologic efficacy against respiratory pathogens, including resistant strains; simple, short-course therapy; and good tolerability and palatability . New antibacterials will continue to have a role in the treatment of RTIs in children, especially where resistance compromises existing therapies.

Klin Lab Diagn, 2004 Jan, (1), 50 - 2
{Selective culture media for Haemophilus bacteria}; Gruber IM et al.; A selectivity factor was specified to the previously developed nutrient medium meant for the cultivation of Haemophilus bacteria--CAE, which is based on the acid hydrolysate of casein, enzyme hydrolysate of animal blood (aminopeptide) and an extract of nutrient yeast . The above nutrient medium containing additionally growth factors V and X, glucose and bacitracin was shown to fit well for the primary sowing of Haemophilus bacteria and it can be used in diagnostic examinations.

FEMS Microbiol Lett, 2004 Mar 12, 232(1), 83 - 7
Antigenic secreted proteins from Haemophilus paragallinarum . A 110-kDa putative RTX protein; Mena-Rojas E et al.; Haemophilus paragallinarum is the causal agent of infectious coryza, an economically important disease for the poultry industry . This bacterium secreted proteins of 25-110 kDa during its growth in brain heart infusion, tryptic soy broth, or Luria-Bertani glucose phosphate media, all lacking serum . Some of these proteins were recognized by sera from chickens experimentally infected with H . paragallinarum . A 110-kDa protein was recognized by a serum pool from convalescent-phase pigs naturally infected with Actinobacillus pleuropneumoniae, and also by a rabbit polyclonal serum against Apx I as well as a rabbit serum against Mannheimia haemolytica leukotoxin, suggesting the presence of an RTX-like protein in H . paragallinarum . H . paragallinarum secreted proteins could be important immunogens in the control of infectious coryza.

Vet Microbiol, 2004 Mar 26, 99(1), 75 - 8
Prior infection with Bordetella bronchiseptica increases nasal colonization by Haemophilus parasuis in swine; Brockmeier SL; The objective of this study was to determine whether Bordetella bronchiseptica would predispose to colonization or disease with Haemophilus parasuis . Three experiments were completed . In the first experiment, three groups of pigs (10 pigs/group) were inoculated intranasally with either B . bronchiseptica, H . parasuis, or with B . bronchiseptica followed by H . parasuis 1 week later . A fourth group of 10 pigs served as a non-infected control group . The second experiment was like the first, except that there were only five pigs per experimental group . The third experiment consisted of only two groups (10 pigs/group), one of which was inoculated intranasally with H . parasuis, whereas the other was inoculated with B . bronchiseptica followed by H . parasuis 1 week later . Pigs were necropsied 1-2 weeks after inoculation with H . parasuis . Mean nasal colonization by H . parasuis was significantly higher in the coinfected groups compared to the groups infected with H . parasuis alone . Pneumonia was present in 9/25 pigs coinfected with B . bronchiseptica and H . parasuis, 5/25 pigs infected with H . parasuis alone, 1/15 pigs infected with B . bronchiseptica alone, and in none of the pigs in the non-inoculated groups . Thus, B . bronchiseptica increased colonization of the upper respiratory tract with H . parasuis.

Vet Microbiol, 2004 Mar 26, 99(1), 1 - 12
Haemophilus parasuis: new trends on diagnosis, epidemiology and control; Oliveira S et al.; Haemophilus parasuis is a commensal organism of the upper respiratory tract of conventional pigs, but under appropriate conditions can invade and cause severe systemic disease, characterized by fibrinous polyserositis, arthritis and meningitis . Factors involved in systemic invasion by H . parasuis remain largely unknown . However, major advances in our knowledge of H . parasuis include (1) development of a species-specific PCR test to detect H . parasuis in clinical samples, (2) study of molecular epidemiology within and between herds, by use of a repetitive element-based PCR, (3) the proposal of an alternative serotyping technique, (4) development and testing of a new in vivo model for pathogenesis and virulence studies, and (5) use of controlled exposure of young pigs to low doses of live, virulent H . parasuis strains to reduce nursery mortality in affected swine herds.

Pediatr Infect Dis J, 2004 Mar, 23(3), 240 - 5
Concomitant administration of a bivalent Haemophilus influenzae type b-hepatitis B vaccine, measles-mumps-rubella vaccine and varicella vaccine: safety, tolerability and immunogenicity; Hesley TM et al.; BACKGROUND: The study was done to verify that concomitant administration of a bivalent Haemophilus influenzae type b-hepatitis B vaccine (Comvax), measles-mumps-rubella vaccine (M-M-RII) and varicella vaccine (Varivax) would be well-tolerated and suitably immunogenic with respect to all vaccine antigens . METHODS: We randomized 822 healthy 12- to 15-month-old children (1:1) to receive concomitant injections of Comvax, M-M-RII and Varivax (concomitant group) or Comvax followed 6 weeks later by injections of M-M-RII and Varivax (nonconcomitant group) . Blood samples taken before and 6 weeks after vaccination were tested for antibodies to all vaccine antigens . RESULTS: Vaccinations were generally well-tolerated . Children in the concomitant and nonconcomitant treatment groups were similar with respect to the safety endpoint of primary interest (16.1 and 19.5%, respectively, had a fever > or =103 degree F rectally at any time within 14 days after either of two clinic visits) . Fifteen serious adverse events were reported (eight in the concomitant group and seven in the nonconcomitant group); all resolved . Elements of two serious adverse events (fever, fever and measles-like rash; both in concomitant group children) were considered possibly related to vaccination . One child was withdrawn from the study because of a nonserious adverse event subsequently judged to be unrelated to vaccination . Similar proportions of vaccinees in the concomitant and nonconcomitant groups developed satisfactory antibody responses to the H . influenzae polysaccharide, polyribosylribitol phosphate (97.8 to 98.7%), hepatitis B surface antigen (99.2 to 100%), measles virus (99.4 to 99.6%), mumps virus (98.4 to 99.2%), rubella virus (100%) and varicella virus (93.2 to 94.6%) . CONCLUSION: Concomitant administration of Comvax, M-M-RII and VARIVAX at the 12- or 15-month clinic visit is one satisfactory way of delivering some of the multiple vaccines indicated during the second year of life.

Carbohydr Res, 2004 Feb 25, 339(3), 529 - 35
Structural analysis of the lipooligosaccharide from the commensal Haemophilus somnus genome strain 129Pt; St Michael F et al.; The structure for the carbohydrate moiety of the lipooligosaccharide (LOS) from the commensal Haemophilus somnus strain 129Pt was elucidated . The structure of the core oligosaccharide and O-deacylated LOS was established by monosaccharide and methylation analyses, NMR spectroscopy and mass spectrometry . The following structure for the major fully extended carbohydrate glycoform of the LOS was determined on the basis of the combined data from these experiments . {Carbohydrate structure: see text} . In the structure Kdo is 3-deoxy-D-manno-octulosonic acid, Hep is L-glycero-D-manno-heptose and PEtn is phosphoethanolamine . Minor amounts of glycoforms containing nonstoichiometric substituents glycine and phosphate at the distal heptose residue were also identified.

Int J Antimicrob Agents, 2004 Feb, 23(2), 129 - 37
Five-day moxifloxacin therapy compared with 7-day co-amoxiclav therapy for the treatment of acute exacerbation of chronic bronchitis; Starakis I et al.; In this randomized, non-blinded study, the efficacy and safety of a 5-day course of moxifloxacin (one 400 mg tablet daily) was compared with that of co-amoxiclav (one 625 mg tablet every 8h) for 7 days, for the treatment of acute exacerbations of chronic bronchitis (AECB) . A total of 162 patients with clear signs of an acute exacerbation of chronic bronchitis were enrolled . Of these, 153 could be studied . Seventy-nine patients were randomized in the moxifloxacin arm and 74 in the co-amoxiclav arm of the study . The primary efficacy parameter was clinical response at 14 days in the evaluable population . A clinical success was classified as resolution or improvement of symptoms . Variables used to assess clinical response included wheeze, cough, dyspnoea, sputum volume, rales and ronchi . The success rate in the moxifloxacin group was 88.6% (70 of 79) and that for co-amoxiclav group was 89.2% (66 of 74) . At follow-up (28-35 days post-treatment), the continued clinical cure rates were 90.0% (63 of 70) for moxifloxacin and 89.4% (59 of 66) for co-amoxiclav . No significant differences were detected between the two groups . A total of 78 pathogenic bacteria were isolated from the sputum samples of the patients, with Moraxella catarrhalis, Haemophilus influenzae and Streptococcus pneumoniae being the most frequently isolated pathogens . The eradication rate at 14 days in the valid patients was similar for both groups, 90.9% (20 of 22) for the moxifloxacin group and 90.0% (18 of 20) for the co-amoxiclav group . Both drugs were well tolerated with no differences in the drug-related adverse effects or the patients withdrawing because of an adverse event . These results and the good spectrum of antibacterial activity make moxifloxacin a promising and also safe alternative for the empirical treatment of AECB.

Drugs Aging, 2004, 21(4), 243 - 58
Acute cough in the elderly: aetiology, diagnosis and therapy; Widdicombe J et al.; Although the frequency of physician consultations and the sale of over-the-counter remedies establish the high prevalence of acute cough in the elderly, epidemiological studies have tended to be imprecise . However, respiratory tract infections in nose, larynx and/or bronchi, either viral or bacterial or both, are by far the commonest cause of acute cough . These are especially frequent and hazardous in the elderly, and community living and institutionalisation may aggravate this problem . A variety of viruses and bacteria have been incriminated, with rhinovirus, influenza and respiratory syncytial viruses, and Streptococcus pneumoniae, Haemophilus influenza and Bordetella pertussis being especially important . Viral infections can readily lead to community-acquired pneumonia . Successful diagnosis should point to successful treatment, and in this respect clinical examination and patient history are paramount, supplemented by chest X-ray, viral and bacterial culture and serological testing . Depending on the results of these tests, specific antibacterial therapy may be called for, although there is dispute as to the merits of antibacterial therapy in cases of uncertain diagnosis . Prevention and prophylaxis for influenza and S . pneumoniae infections are now commendably routine in the elderly, especially those in communities . Treatment, as well as the use of antibacterials, may also be directed against the inflammatory and infective processes in the airways . Non-specific antitussive therapy is common and usually highly desirable to prevent the adverse effects of repeated coughing . There have been few advances in antitussive therapy in recent years, opioids and dextromethorphan being the most commonly used agents; they act centrally on the brainstem, but also have a large placebo effect . However they work, they are much appreciated by patients and their partners . Moreover, striking advances in our understanding of the peripheral sensory and central nervous pathways of the cough reflex in recent years should soon lead to a new and more specific choice of agents to inhibit cough.

In Vivo, 2004 Jan-Feb, 18(1), 67 - 71
Etiological agents of lower respiratory tract infections in Japanese children; Numazaki K et al.; To investigate the etiology of pediatric community-acquired pneumonia and bronchitis, we conducted a prospective, population-based study covering the total population < 15 years of age in 16 municipalities in Hokkaido, Japan, during the period of April 2000 to March 2001 . Chest radiographs were available for all cases (n = 921; 398 as pneumonia and 523 as bronchitis) and paired sera for serologic assays were available for more than half of the cases . The following specimens were also collected: nasopharyngeal swabs for viral, bacteriological, mycoplasmal and chlamydial studies, blood for serology and blood culture . The children were then followed-up on days 3, 7 and 14 . Specific infecting organisms were identified in a total of 853 (92.6%) out of 921 patients (398 cases of pneumonia and 523 cases of bronchitis) including 205 with mixed infection as follows: Mycoplasma pneumoniae, 252 (274%) patients; respiratory syncytial (RS) virus, 188 (20.4%); influenza A virus, 110 (11.9%); Streptococcus pneumoniae, 95 (10.3%); Haemophilus influenzae, 90 (9.8%); Haemophilus parainfluenzae, 35 (3.8%); Staphylococcus aureus, 29 (3.1%); adenovirus, 27 (2.9%); Moraxella catarrhalis, 12 (1.3%); Pseudomonas aeruginosa , 7 (0.8%); Chlamydia pneumoniae, 6 (0.7%); and other agents, 2 (0.2%) . Mycoplasma infections were seen even in patients less than 5 years and RS and influenza A virus infections in patients more than 5 years of age . The importance of M . pneumoniae and RS virus in the etiology of lower respiratory infections in Japanese children was confirmed.

Arch Med Res, 2004 Mar-Apr, 35(2), 126 - 33
Invasive Haemophilus influenzae type b infections in children in Paraguay; Basualdo W et al.; BACKGROUND: In Paraguay, as in most Latin American countries, data on the epidemiology and clinical characteristics of Haemophilus influenzae type b (Hib) diseases are scarce and incomplete . METHODS: To address this issue, we performed a retrospective analysis of 102 patients admitted to the Instituto de Medicina Tropical, a referral hospital in Asuncion, Paraguay, between January 1991 and September 1995 with diagnosis of invasive Hib infection . This study included patients 15 years of age and under-identified with positive cultures for Hib in blood, cerebrospinal fluid, or other sterile body fluids . RESULTS: Eighty three (81%) patients presented with meningitis as principal focus of infection with median age of 9 months . Forty five (54%) patients with Hib meningitis were <12 months of age and 20 (24% of total cases) were <6 months of age . Overall mortality rate of meningitis was 13% . Of 11 patients who died, 10 (91%) were <12 months of age (p <0.02) . Risk for mortality was correlated with presence of coma during admission (p <0.007) and CSF glucose level of <10 mg/dL (p <0.05) . Severe sequelae such as bilateral hearing loss, hydrocephalus, and mental retardation were observed in 39% (28/72) of surviving patients, of whom 18 (51%) patients were <12 months of age (p <0.02) . Thirty percent of isolated strains of Hib were resistant to ampicillin, 20% were resistant to chloramphenicol, and 10% to both drugs . CONCLUSIONS: This information provides evidence concerning the importance of continued support for Hib vaccine supplies in immunization programs in countries with limited resources such as Paraguay.

Eur J Biochem, 2004 Mar, 271(5), 941 - 53
Characterization of novel structural features in the lipopolysaccharide of nondisease associated nontypeable Haemophilus influenzae; Landerholm MK et al.; Nontypeable Haemophilus influenzae (NTHi) is a common commensal of the human upper respiratory tract and is associated with otitis media in children . The structures of the oligosaccharide portions of NTHi lipopolysaccharide (LPS) from several otitis media isolates are now well characterized but it is not known whether there are structural differences in LPS from colonizing, nondisease associated strains . Structural analysis of LPS from nondisease associated NTHi strains 11 and 16 has been achieved by the application of high-field NMR techniques, ESI-MS, ESI-MSn, capillary electrophoresis coupled to ESI-MS, composition and linkage analyses on O-deacylated LPS and core oligosaccharide material . This is the first study to report structural details on LPS from strains taken from the nasopharynx from healthy individuals . Both strains express identical structures and contain the common element of H . influenzae LPS, L-alpha-D-Hepp-(1-->2)-{PEtn-->6}-L-alpha-D-Hepp-(1-->3)-{beta-D-Glcp-(1-->4)}-L-alpha-D-Hepp-(1-->5)-{PPEtn-->4}-alpha-Kdop-(2-->6)-lipid A, in which each heptose is elongated by a single hexose residue with no further oligosaccharide extensions . In the major Hex3 glycoform, the terminal Hepp residue (HepIII) is substituted at the O-2 position by a beta-D-Galp residue and the central Hepp residue (HepII) is substituted at O-3 by a alpha-D-Glcp residue . Notably, the strains express two phosphocholine (PCho) substituents, one at the O-6 position of alpha-D-Glcp and the other at the O-6 position of beta-D-Galp . Major acetylation sites were identified at O-4 of Gal and O-3 of HepIII . Additionally, both strains express glycine, and strain 11 also expresses detectable amounts of N-acetylneuraminic acid.

Clin Exp Immunol, 2004 Mar, 135(3), 474 - 7
Natural materno-fetal transfer of antibodies to PspA and to PsaA; Baril L et al.; PspA and PsaA are Streptococcus pneumoniae surface proteins and potential pneumococcal vaccine antigens . The aim of this study was to characterize the transplacental transfer of antibodies to PspA and to PsaA . Paired mother and cord blood sera were obtained at delivery from 28 women . Concentrations of antibodies against PspA, PsaA, tetanus toxoid (vaccine-induced antibodies) and P6-outer membrane protein (OMP) of nontypeable Haemophilus influenzae were determined by ELISA . Antibodies to PspA of the IgG, IgG1 and IgG2 antibodies were also determined . The geometric mean percentage (GM%) of the paired infant:mother antibody were calculated . Results: The GM% of the infant:mother antibody concentrations against PspA, PsaA and P6-OMP antibodies were 64.7% (3.3 micro g/ml in infants vs . 5.1 micro g/ml in mothers), 50.4% (6.8 micro g/ml vs . 13.5 micro g/ml) and 66.7% (5.6 micro g/ml vs . 8.4 micro g/ml), respectively; the GM% of antibodies against tetanus toxoid was 104.5% (4.6 micro g/ml vs . 4.4 micro g/ml) . Transplacental transfer of IgG1 was more efficient than that of IgG2 (approximately 120%vs . 65%) . A transplacental transfer of antibodies to PspA and to PsaA exist . Moreover, these data suggest an active placental transfer of IgG1 antibodies to PspA since the concentration of these antibodies were consistently higher in cord sera than in the mother's sera.

Microb Drug Resist, 2003 Winter, 9(4), 337 - 44
Clonal analysis of Streptococcus pneumoniae nonsusceptible to penicillin at day-care centers with index cases, in a region with low incidence of resistance: emergence of an invasive type 35B clone among carriers; Henriqus Normark B et al.; The nasopharyngeal carriage rate of potential respiratory pathogens was studied in 36 index children with a pneumococci nonsusceptible to penicillin (PNSP), in 595 healthy children, and in 123 personnel at 16 day-care centers (DCCs) with index cases in the Stockholm area, an urban area with a low incidence of antibiotic resistant pneumococci, during the winter of 1997-1998 . The spread and clonality of PNSP, Haemophilus influenzae and Moraxella catarrhalis, were studied by analyzing antibiotic susceptibility and serotype, and for PSNP also by using pulsed-field electrophoresis (PFGE) and multilocus sequence typing (MLST) . In contrast to the low carriage rate found among the adult contacts (2%), 40% of the children harbored pneumococci, of which 20% were PNSP . Nasopharyngeal colonization decreased with age . The 49 PNSP isolates consisted of 20 clones, of which 10 could be identified in more than one child attending the same or different DCCs . In five DCCs, we observed a spread of PNSP from the index case . A novel PNSP clone of type 35B, found to cause invasive disease in several states in the United States, was found to emerge among several carriers at two DCCs . A high proportion of PNSP isolates were multiresistant to antibiotics (34%), which has implications for treatment regimens, even in a country like Sweden where the proportion of PNSP currently is low (3-4%) . One PNSP clone of type 9V found among the carriers, has been shown to cause invasive disease in Sweden as well as in other countries, suggesting that one reason for the occurrence of invasive PNSP clones may be their ability to colonize and spread among healthy carriers . Other internationally spread antibiotic resistant pneumococcal clones found were of types 9V, 19F, and 23F.

Jpn J Antibiot, 2003 Dec, 56(6), 681 - 90
{Antibacterial activity of oral Cephems against various clinically isolated strains}; Oshiro T et al.; We determined the antibacterial activities of oral Cephems against isolated from the patients with the respiratory infections, the urinary tract infections, and infections in the obstetrics field of an adult and a child, during the period from 2002 to 2003; Streptococcus pyogenes, Streptococcus pneumoniae, Haemophilus influenzae, Branhamella catarrhalis, Klebsiella pneumoniae and Escherichia coli of 40 strains of each, and Peptostreptococcus spp . 22 strains . S . pneumoniae and H . influenzae strains that resistant is regarded were collected mainly, penicillin-intermediate S . pneumoniae (PISP), penicillin-resistant S . pneumoniae (PRSP) and beta-lactamase negative ampicillin-resistant H . influenzae (BLNAR) strains . The MICs of Cephems except cefaclor (CCL) were < or = 0.03 microgram/mL against all strains of S . pyogenes . The MICs of cefteram (CFTM) and cefditoren (CDTR) were < or = 0.0125 microgram/mL activity against 7 strains penicillin-susceptible S . pneumoniae (PSSP) . However the MIC90s of cefditoren (CDTR) was 1 microgram/mL, cefteram (CFTM), and cefcapene (CFPN) were 2 micrograms/mL against PISP and PRSP, were higher than those of other drugs, but showed slightly higher than PSSP . The MIC90s of Cephems . were 0.5-4 micrograms/mL against strains of E . coli . The MIC90s of CFTM was 0.5 microgram/mL, and CDTR, CFPN were 1 microgram/mL against E . coli were higher than those of other drugs . The four strains of E . coli however were highly-resistant which MIC90s of CCL were more than 32 micrograms/mL were obtains . Furthermore it is necessary to pay much attention to the trend of resistant such as E . coli of Cephems . Although all strains showed resistant to AMPC, MIC90 of Cephems were 0.25-1 microgram/mL, good activities against K . pneumoniae . Against beta-lactamase negative ampicillin-susceptible H . influenzae (BLNAS) 23 strains the MIC90s of CCL and other Cephems were 64 micrograms/mL and 0.25-8 micrograms/mL . The MIC90s of CDTR and CFTM were < or = 1 microgram/mL of BLNAR (15 strains) . However there of CFDN and CPDX were 8 micrograms/mL and CCL were > or = 16 micrograms/mL . Two strains which were produced beta-lactamase were highly--ABPC resistant . Although B . catarrhalis all strains were produced beta-lactamase and Cephems except for CCL showed better susceptibility than AMPC . The MIC90s of Cephems were 0.25-2 micrograms/mL against Peptostreptococcus spp.

J Pediatr Health Care, 2004 Mar-Apr, 18(2), 72 - 6
Acute sinusitis in children: diagnosis and management; Leung AK et al.; Acute sinusitis is a common problem in children that is often overlooked . The pathophysiology is related to obstruction of the sinus ostia and mucociliary dysfunction . The predominant pathogens include Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis . The diagnosis of acute sinusitis is based on persistent or severe upper respiratory tract symptoms . The routine use of roentgenograms to confirm the diagnosis of uncomplicated sinusitis is not recommended for children 6 years or younger but may be considered for children older than 6 years . Antimicrobial therapy is the cornerstone of management and amoxicillin is the drug of choice for most cases of uncomplicated sinusitis.

Ophthalmic Res, 2004 Jan-Feb, 36(1), 62 - 9
Quantitative determination of histamine in tears during conjunctivitis by a novel HPLC method; Venza I et al.; The histamine content of tears of healthy sex- and age-matched subjects and patients affected by allergic or nonallergic inflammatory ocular diseases was determined through a new competitive reverse-phase high-performance liquid chromatography (HPLC) method . Tear samples from 50 healthy subjects, 30 patients affected by seasonal allergic conjunctivitis, 12 patients with bacterial conjunctivitis associated with Haemophilus influenzae and 8 patients with bacterial conjunctivitis associated with Streptococcus pneumoniae were analyzed for histamine concentration by O-phthaldialdehyde precolumn derivatization-based HPLC . In physiological conditions, the tear histamine content was low (2.26 ng/ml) and did not vary in relation to age and sex . Histamine levels were significantly higher in all the patients studied, to a greater extent in those affected by allergic (23.61 ng/ml) or Haemophilus influenzae-associated (21.53 ng/ml) conjunctivitis .

Ann Trop Paediatr, 2004 Mar, 24(1), 95 - 101
Five children with purulent pericarditis and review of the literature; Kabbani MS et al.; Purulent pericarditis in children is a life-threatening disease that requires early diagnosis and immediate intervention . This cardiac emergency is rarely seen in the western world . However, cases of purulent pericarditis are still being reported in developing countries . We describe our experience with five cases of purulent pericarditis in children seen between 1998 and 2002 . Haemophilus influenzae bacteria were isolated in all except one case . With active management, all five children survived.

J Clin Microbiol, 2004 Mar, 42(3), 1185 - 91
Laboratory detection of Haemophilus influenzae with decreased susceptibility to nalidixic acid, ciprofloxacin, levofloxacin, and moxifloxacin due to GyrA and ParC mutations; Perez-Vazquez M et al.; The detection of clinical isolates with decreased fluoroquinolone susceptibilities and a resistance mechanism is of epidemiological and clinical interest . We studied the susceptibilities of 62 clinical isolates and 2 American Type Culture Collection reference strains of Haemophilus influenzae to ciprofloxacin, levofloxacin, moxifloxacin, and nalidixic acid by the microdilution and disk diffusion methods . The ciprofloxacin MICs for 34 of the isolates were >/=0.12 micro g/ml (range, 0.12 to 32 micro g/ml), and the ciprofloxacin MICs for 28 matched control isolates were </=0.06 micro g/ml . In addition, we sequenced the quinolone resistance-determining regions (QRDRs) of gyrA and parC of all strains . The log(2) MICs of all quinolones were plotted against the inhibition zone diameters . The MICs and inhibition zone diameters selected to screen for the resistance mechanism were based on the susceptibility distribution data and the presence or absence of amino acid changes in the QRDRs of GyrA and ParC . Strains for which ciprofloxacin MICs were </=0.06 micro g/ml, levofloxacin and moxifloxacin MICs were </=0.03 micro g/ml, and nalidixic acid MICs were </=2.0 micro g/ml lacked modifications in the QRDR of GyrA . In contrast, all strains for which ciprofloxacin, levofloxacin, and moxifloxacin MICs were >/=0.5 micro g/ml and the vast majority of those for which nalidixic acid MICs were >/=32 micro g/ml exhibited amino acid changes in GyrA and ParC . Nalidixic acid and the other three fluoroquinolones studied could be used to screen H . influenzae isolates for the detection of decreased susceptibilities to quinolones due to the acquisition of two amino acid changes in the QRDRs of GyrA and ParC (sensitivity, >95%; specificity, >80%).

Ann Trop Med Parasitol, 2004 Jan, 98(1), 65 - 70
The role of the polymerase chain reaction in the diagnosis of bacterial meningitis in Vietnam; Freeman HR et al.; Bacterial meningitis remains an important cause of morbidity and mortality in Vietnam . Diagnosis is hampered by the ready availability of antibiotics in the community, leading to late presentation, masked clinical signs, and poor organism detection during the microscopical examination and culture of cerebrospinal fluid (CSF) . In order to improve organism detection at the Hospital for Tropical Diseases in Ho Chi Minh City, a diagnostic PCR-based protocol was developed . This protocol was followed in the investigation of CSF samples from 36 patients with clinical signs of bacterial meningitis . Each sample was first tested in a semi-nested PCR using primers for the 16sRNA gene common to all bacteria . The products of this reaction were then amplified using a 16sru8 primer and primers specific for Neisseria meningitidis, Haemophilus influenzae or Streptococcus spp . The samples found positive for Streptococcus were further investigated in a nested PCR using primers specific for the pneumolysin gene of S . pneumoniae . The sensitivity of detection was increased from 36% with culture to 44% with PCR . Although the sample size was small, the results indicate that PCR would be a feasible and useful adjunct in the diagnosis of bacterial meningitis, particularly in areas where community antibiotic use is common.

Mutat Res, 1976 Jun, 35(2), 199 - 205
Mutation of Haemophilus influenzae transforming DNA in vitro with near-ultraviolet radiation: action spectrum; Cabrera-Juarez E et al.; Mutations were produced in purified transforming DNA from Haemophilus influenzae by near-UV radiation and were assayed as mutants among cells transformed with irradiated DNA . The maximum efficiency of mutation induction was at around 334 nm, and the efficiency dropped off steeply at lower and higher wavelengths . The difference between the action spectrum for mutation and that for the oxygen-independent inactivation of transforming DNA, which had a shoulder at 365 nm, indicates that there are different lesions involved in the inactivating and mutagenic effects of near-UV . The presence of histidine during irradiation enhanced the mutagenic effect at 334 and 365 nm, although it protected against inactivation at 365 nm . The effective near-UV wavelengths for in vitro mutation are to some extent the same as the effective wavelengths for mutation in vivo reported previously . These findings indicate that mutations are produced in vivo by near-UV with DNA as the primary target molecule rather than by a secondary non-photochemical reaction between DNA and some other cell component.

Proc Natl Acad Sci U S A, 2004 Mar 9, 101(10), 3563 - 8 Epub 2004 Mar 01.
Synergistic activation of NF-kappaB by nontypeable Haemophilus influenzae and tumor necrosis factor alpha; Watanabe T et al.; Nontypeable Haemophilus influenzae (NTHi) is an important human pathogen causing otitis media in children and exacerbation of chronic obstructive pulmonary disease in adults . Like most other bacterial infections, NTHi infections are also characterized by inflammation, which is mainly mediated by cytokines and chemokines such as tumor necrosis factor alpha (TNF-alpha) . Among a variety of transcription regulators, NF-kappaB has been shown to play a critical role in regulating the expression of large numbers of genes encoding inflammatory mediators . In review of the current studies on NF-kappaB regulation, most of them have focused on investigating how NF-kappaB is activated by a single inducer at a time . However, in bacteria-induced inflammation in vivo, multiple inducers including both exogenous and endogenous mediators are present simultaneously . A key issue that has yet to be addressed is whether the exogenous inducers such as NTHi and the endogenous factors such as TNF-alpha activate NF-kappaB in a synergistic manner . We show that NTHi and TNF-alpha, when present together, synergistically induce NF-kappaB activation via two distinct signaling pathways: NF-kappaB translocation-dependent and -independent pathways . The NF-kappaB translocation-dependent pathway involves NF-kappaB-inducing kinase-IkappaB kinase beta/gamma-dependent phosphorylation and degradation of IkappaBalpha, whereas the NF-kappaB translocation-independent pathway involves mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase kinase kinase 1-dependent activation of MAPK kinase 3/6-p38 MAPK pathway . In addition, the same signaling pathways are also involved in synergistic induction of TNF-alpha, IL-1beta, and IL-8 . These studies should deepen our understanding of the molecular mechanisms underlying the combinatorial regulation of inflammation and lead to development of therapeutic strategies for NTHi-induced infections.

Genome Res, 2004 Mar, 14(3), 343 - 53
Comparative analysis of protein domain organization; Ye Y et al.; We have developed a set of graph theory-based tools, which we call Comparative Analysis of Protein Domain Organization (CADO), to survey and compare protein domain organizations of different organisms . In the language of CADO, the organization of protein domains in a given organism is shown as a domain graph in which protein domains are represented as vertices, and domain combinations, defined as instances of two domains found in one protein, are represented as edges . CADO provides a new way to analyze and compare whole proteomes, including identifying the consensus and difference of domain organization between organisms . CADO was used to analyze and compare >50 bacterial, archaeal, and eukaryotic genomes . Examples and overviews presented here include the analysis of the modularity of domain graphs and the functional study of domains based on the graph topology . We also report on the results of comparing domain graphs of two organisms, Pyrococcus horikoshii (an extremophile) and Haemophilus influenzae (a parasite with reduced genome) with other organisms . Our comparison provides new insights into the genome organization of these organisms . Finally, we report on the specific domain combinations characterizing the three kingdoms of life, and the kingdom "signature" domain organizations derived from those specific domain combinations.

J Infect Chemother, 2004 Feb, 10(1), 31 - 6
Value of washed sputum gram stain smear and culture for management of lower respiratory tract infections in children; Cao LD et al.; To date, the technique of washed sputum examinations has not been widely used in the clinical management of lower respiratory tract infections in children . A total of 224 sputum samples from 125 pediatric patients with lower respiratory tract infections were collected for washed sputum Gram stain smears and cultures . The results with these methods were compared to find correlation rates . The value of washed sputum cultures was assessed by examining the clinical responses of the patients who received antibiotic therapies instituted on the basis of the sputum culture results . Isolation rates of Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis, and Staphylococcus aureus were 22.4%, 9.4%, 4.9%, and 0.4%, respectively . For the prediction of H . influenzae, S . pneumoniae, and M . catarrhalis, the sensitivities of the washed sputum Gram stain smears compared with the culture method were 86.0%, 81.0%, and 90.9%, respectively . The specificities of the washed sputum Gram stain smear technique were 94.8%, 97.5%, and 98.1%, respectively . Overall, the sensitivity and specificity of the washed sputum Gram stain smear method were 85.5% and 87.2%, respectively . S . aureus was isolated from only one specimen; and washed sputum Gram stain smear estimation was correlated with the culture result . On the basis of the washed sputum culture results, appropriate antibiotic therapies were instituted for 93.3% of the patients with acute lower respiratory tract infections . This study suggests that the techniques of washed sputum Gram stain smear and culture are valuable and should be encouraged in clinical practice for the management of lower respiratory tract infections in children.

Zhonghua Jie He He Hu Xi Za Zhi, 2004 Jan, 27(1), 27 - 30
{Prevalence of atypical pathogens in adult patients with community-acquired pneumonia in Beijing}; Liu YN et al.; OBJECTIVE: To study the pathogens in community-acquired pneumonia (CAP), especially the prevalence of atypical pathogens . METHODS: A prospective study was performed on 103 consecutive adult patients with CAP between November 2001 and June 2002 . Antibodies of the paired serum samples to Mycoplasma pneumoniae, Legionella pneumophilia, and Chlamydia pneumoniae were detected . The P1 adhesion gene of Mycoplasma pneumoniae and the 16S ribosome gene specific for Chlamydia pneumoniae were detected with polymerase chain reaction (PCR) . Legionella antigen in urine was detected using enzyme immunoassay (EIA) method . Sputum samples were collected for culture, and bacteria were isolated and identified using conventional methods . RESULTS: The etiology of CAP was identified in 50 (48.5%) patients . The distribution of causal agents was as follows: Mycoplasma pneumoniae in 23 (22.3%) cases, Legionella pneumophilia 3 (2.9%), Chlamydia pneumoniae 2 (1.9%), streptococcus pneumoniae 12 (11.7%), Haemophilus influenzae 9 (8.7%), and Klebsiella pneumoniae 7(6.8%) . In 6 patients (5.8%) more than one causal agent were found, among them Mycoplasma pneumoniae was found in 5 cases with mixed infections . CONCLUSIONS: Atypical pathogens especially Mycoplasma pneumoniae have an important role in CAP . Streptococcus pneumoniae and Haemophilus influenzae remain the most common bacteria, and mixed infection should not be ignored.

Eur J Med Chem, 2004 Feb, 39(2), 135 - 40
Synthesis and antimicrobial properties of 2-(benzylidene-amino)-benzo{d}isothiazol-3-ones; Zani F et al.; The in vitro antimicrobial activity of 2-amino-benzo{d}isothiazol-3-one and of several 2-arylideneamino derivatives carrying in the second position a substituted or unsubstituted aromatic ring or an arylalkenylidene moiety was determined by the broth dilution method against several strains selected to define their spectrum and potency . All the compounds demonstrated good antibacterial properties against Bacillus subtilis, streptococci, enterococci and staphylococci including penicillin-resistant clinical isolates . Several compounds showed excellent inhibitory properties against Streptococcus pyogenes, which is the most sensitive microorganism tested . Many benzisothiazolones exhibited good activity against Gram-negative Haemophilus influenzae . As regards antifungal activity, several of the tested compounds inhibited Saccharomyces cerevisiae at concentrations of 3-6 microg ml-1 . In all cases the parent 2-amino-benzo{d}isothiazol-3-one was the most effective agent, with minimum inhibitory concentration (MIC) values ranging from 0.07 to 6 microg ml-1 . The results obtained indicate that most of these compounds are wide-spectrum antimicrobial substances and promising agents against penicillin-resistant staphylococci.

Cancer Immunol Immunother, 2004 Jul, 53(7), 642 - 50 Epub 2004 Feb 17.
Phase I/II trial of immunogenicity of a human papillomavirus (HPV) type 16 E7 protein-based vaccine in women with oncogenic HPV-positive cervical intraepithelial neoplasia; Hallez S et al.; Purpose: Infection with oncogenic human papillomavirus (HPV) and HPV-16 in particular is a leading cause of anogenital neoplasia . High-grade intraepithelial lesions require treatment because of their potential to progress to invasive cancer . Numerous preclinical studies have demonstrated the therapeutic potential of E7-directed vaccination strategies in mice tumour models . In the present study, we tested the immunogenicity of a fusion protein (PD-E7) comprising a mutated HPV-16 E7 linked to the first 108 amino acids of Haemophilus influenzae protein D, formulated in the GlaxoSmithKline Biologicals adjuvant AS02B, in patients bearing oncogenic HPV-positive cervical intraepithelial neoplasia (CIN) . Methods: Seven patients, five with a CIN3 and two with a CIN1, received three intramuscular injections of adjuvanted PD-E7 at 2-week intervals . Three additional CIN1 patients received a placebo . CIN3 patients underwent conization 8 weeks postvaccination . Cytokine flow cytometry and ELISA were used to monitor antigen-specific cellular and antibody responses from blood taken before and after vaccine or placebo injection . Results: Some patients had preexisting systemic IFN-gamma CD4+ (1/10) and CD8+ (5/10) responses to PD-E7 . Vaccination, not placebo injection, elicited systemic specific immune responses in the majority of the patients . Five vaccinated patients (71%) showed significantly increased IFN-gamma CD8+ cell responses upon PD-E7 stimulation . Two responding patients generated long-term T-cell immunity toward the vaccine antigen and E7 as well as a weak H . influenzae protein D (PD)-directed CD4+ response . All the vaccinated patients, but not the placebo, made significant E7- and PD-specific IgG . Conclusions: The encouraging results obtained from this study performed on a limited number of subjects justify further analysis of the efficacy of the PD-E7/AS02B vaccine in CIN patients .

Respirology, 2004 Mar, 9(1), 120 - 4
Hospital-acquired pneumonia in general wards of a Japanese tertiary hospital; Ohi H et al.; OBJECTIVE: Hospital-acquired pneumonia (HAP) is an important nosocomial disorder because both its mortality and morbidity are high . The Japanese Respiratory Society (JRS) has proposed a new guideline for the diagnosis and treatment of HAP . Clinical investigations of HAP were conducted to assess the current status of HAP in accordance with this guideline . METHODOLOGY: Inpatients who developed HAP in the Second Department of Internal Medicine, Nagasaki University Hospital, a tertiary hospital in Japan, were evaluated . The incidence of HAP was 1.86% (59 cases in 3176 admissions) from 1996 to 2000 . In particular, the severity rating of HAP for 46 patients in accordance with the JRS guidelines, was assessed . RESULTS: The mortality rate was 6.5% at this hospital, which is less than that reported by other investigators in the field . The frequency of bacteria isolated from clinical specimens was as follows: Haemophilus influenzae (20%), Pseudomonas aeruginosa (16.7%), and methicillin-resistant Staphylococcus aureus (13.3%) . Response rates were stratified by first-line medications and the results were as follows: penicillins (33.3%), third-generation cephems (44.4%), and carbapenems (66.7%) . CONCLUSIONS: Based on the results presented here, the severity rating proposed in this guideline for HAP appears reasonable, as the response rate for the first-line antibiotic was better in Group II than in Groups III and IV . It will be important to analyse 'clinical outcome' in accordance with this guideline.

Drugs Aging, 2004, 21(3), 167 - 86
New developments in antibacterial choice for lower respiratory tract infections in elderly patients; Ferrara AM et al.; Elderly patients are at increased risk of developing lower respiratory tract infections compared with younger patients . In this population, pneumonia is a serious illness with high rates of hospitalisation and mortality, especially in patients requiring admission to intensive care units (ICUs) . A wide range of pathogens may be involved depending on different settings of acquisition and patient's health status . Streptococcus pneumoniae is the most common bacterial isolate in community-acquired pneumonia, followed by Haemophilus influenzae, Moraxella catarrhalis and atypical pathogens such as Chlamydia pneumoniae, Legionella pneumophila and Mycoplasma pneumoniae . However, elderly patients with comorbid illness, who have been recently hospitalised or are residing in a nursing home, may develop severe pneumonia caused by multidrug resistant staphylococci or pneumococci, and enteric Gram-negative bacilli, including Pseudomonas aeruginosa . Moreover, anaerobes may be involved in aspiration pneumonia . Timely and appropriate empiric treatment is required in order to enhance the likelihood of a good clinical outcome, prevent the spread of antibacterial resistance and reduce the economic impact of pneumonia . International guidelines recommend that elderly outpatients and inpatients (not in ICU) should be treated for the most common bacterial pathogens and the possibility of atypical pathogens . The algorithm for therapy is to use either a selected beta-lactam combined with a macrolide (azithromycin or clarithromycin), or to use monotherapy with a new anti-pneumococcal quinolone, such as levofloxacin, gatifloxacin or moxifloxacin . Oral (amoxicillin, amoxicillin/clavulanic acid, cefuroxime axetil) and intravenous (sulbactam/ampicillin, ceftriaxone, cefotaxime) beta-lactams are agents of choice in outpatients and inpatients, respectively . For patients with severe pneumonia or aspiration pneumonia, the specific algorithm is to use either a macrolide or a quinolone in combination with other agents; the nature and the number of which depends on the presence of risk factors for specific pathogens . Despite these recommendations, clinical resolution of pneumonia in the elderly is often delayed with respect to younger patients, suggesting that optimisation of antibacterial therapy is needed . Recently, some new classes of antibacterials, such as ketolides, oxazolidinones and streptogramins, have been developed for the treatment of multidrug resistant Gram-positive infections . However, the efficacy and safety of these agents in the elderly is yet to be clarified . Treatment guidelines should be modified on the basis of local bacteriology and resistance patterns, while dosage and/or administration route of each antibacterial should be optimised on the basis of new insights on pharmacokinetic/pharmacodynamic parameters and drug interactions . These strategies should be able to reduce the occurrence of risk factors for a poor clinical outcome, hospitalisation and death.

Can J Vet Res, 2004 Jan, 68(1), 71 - 5
Computer-based analysis of Haemophilus parasuis protein fingerprints; Oliveira S et al.; The present study aimed to compare the whole-cell protein profiles of Haemophilus parasuis field isolates by using a computer-based analysis, and evaluate the relationship between polyacrylamide gel electrophoresis (PAGE) type and virulence potential based on isolation site . A dendrogram clustering isolates with similar protein profiles was generated . Haemophilus parasuis isolates were grouped into 2 major PAGE type groups . The PAGE type II isolates were characterized by the presence of major proteins with molecular weights varying from between 36 and 38 kDa and included 90.7% of the isolates recovered from systemic sites, such as pleura, pericardium, peritoneum, lymph nodes, joints, and brain . Isolates classified as PAGE type I were characterized by the absence of this group of proteins and included 83.4% of the isolates recovered from the upper respiratory tract of healthy animals . The present study further corroborates the existence of a unique group of major proteins in potentially virulent H . parasuis isolates.

J Pediatr (Rio J), 2004 Jan-Feb, 80(1), 41 - 8
{Prevalence of bacteria in children with otitis media with effusion}; Pereira MB et al.; OBJECTIVES: 1) To determine the prevalence of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis in middle ear effusions of children with otitis media with effusion undergoing myringotomy; 2) to compare the results obtained by culture and PCR; and 3) to determine the susceptibility of bacterial isolates to penicillin . METHODS: We analyzed 128 middle ear effusion specimens from 75 children (age = 11 months to 10 years; mean = 34.7 months) . Patients with recurrent otitis media had documented middle ear effusion for >/= 6 weeks, and chronic otitis media with effusion for >/= 3 months . The patients had no signs of acute otitis media or respiratory tract infection and were not on antibiotic therapy . Aspiration was done through tympanocentesis with an Alden-Senturia trap . Bacteriological studies were initiated less than 15 minutes after specimen collection . Part of the sample was stored at -20oC for later multiplex PCR analysis . Statistical analysis employed McNemar's (Chi2) test . RESULTS: Bacteria were cultured in 32 (25.1%) out of 128 samples and the pathogens under investigation were found in 25 (19.6%) . PCR was positive for bacteria in 73 (57.0%) specimens: 50 (39.1%) for H . influenzae, 16 (12.5%) for S . pneumoniae, and 13 (10.2%) for M . catarrhalis . All the culture-positive samples were PCR-positive, but 48 (65.7%) of the PCR-positive specimens were culture-negative . PCR was significantly more sensitive than culture (p < 0.01) to identify bacteria . Resistance to penicillin was as follows: M . catarrhalis = 100%; S . pneumoniae = 62.5% and H . influenzae = 23% of the isolates . CONCLUSIONS: The prevalence of bacteria in otitis media with effusion in a group of Brazilian children was similar to that reported for other countries . H . influenzae was the most frequent microorganism observed . This suggests that bacteria may play a role in the pathogenesis of otitis media with effusion . In addition, PCR was more sensitive to detect bacteria in middle ear effusion as compared to conventional culture methods . Penicillin resistance was similar to that reported for other countries for pneumococci and moraxella, but beta-lactamase production by H . influenza was lower than that reported for other countries.

Infez Med, 1996, 4(2), 64 - 73
{Emerging pathogens in lower respiratory tract infections}; Paradisi F et al.; Pneumonia is one of the most important causes of death in industrialized countries where it is the infection with the overall highest mortality rate . In contrast to bacteremia, in which the cause of infection is readily established, the determination of the cause of acute pneumonia remains problematic . About half the patients requiring admission to the hospital do not have the cause of pneumonia diagnosed . The emergence of newer pathogens and the recognition of new presentations of older pathogens, however, have made it apparent that it will always be necessary to pursue aggressively an etiologic diagnosis in patients with pneumonia . This article briefly reviews some of new pathogens of lower respiratory tract infections that have become very important in the last years for their capability to determine severe clinical patterns, for their high frequency in some high-risk patient groups and for difficult diagnosis or treatment . Moreover we present new facets of the "old pathogens" of community-acquired pneumonia . Particularly we underline major changes occurred in the antibiotic-susceptibility of Streptococcus pneumoniae and Haemophilus influenzae.

J Immunol, 2004 Mar 1, 172(5), 3305 - 18
Human memory B cells transferred by allogenic bone marrow transplantation contribute significantly to the antibody repertoire of the recipient; Lausen BF et al.; The bone marrow is an important source of Abs involved in long-term protection from recurrence of infections . Allogenic bone marrow transplantation (BMT) fails to restore this working memory . Attempts to overcome this immunodeficiency by immunization of the donor have not been very successful . More needs to be known about transfer of B cell memory by BMT . We tracked memory B cells from the donor to the recipient during BMT of a girl with leukocyte adhesion deficiency . Vaccination of her HLA-identical sibling donor 7 days before harvest induced Haemophilus influenzae type b (Hib) capsular polysaccharide (HibCP)-specific B cells readily detectable in marrow and blood . BMT did not lead to spontaneous production of HibCP Abs, but the recipient responded well to booster immunizations 9 and 11 mo after BMT . HibCP-specific B cells were obtained 7 days after the vaccinations, and their V(H) genes were sequenced and analyzed for rearrangements and unique patterns of somatic hypermutations identifying clonally related cells . Ninety (74%) of 121 sequences were derived from only 16 precursors . Twelve clones were identified in the donor, and representatives from all of them were detected in the recipient where they constituted 61 and 68% of the responding B cells after the first and second vaccinations, respectively . No evidence for re-entry of memory clones into the process of somatic hypermutation was seen in the recipient . Thus, memory B cells were transferred from the donor, persisted for at least 9 mo in the recipient, and constituted the major part of the HibCP-specific repertoire.

J Antimicrob Chemother, 2004 Apr, 53(4), 664 - 8 Epub 2004 Feb 18.
In vitro antibacterial activity of the peptide deformylase inhibitor BB-83698; Lofland D et al.; OBJECTIVES: BB-83698 is a peptide deformylase inhibitor currently in clinical trials in Europe . The purpose of this study was to provide additional susceptibility data from clinical isolates, including drug-resistant strains . METHODS: The in vitro activities of BB-83698 and comparators were determined against 281 streptococci, 154 Staphylococcus aureus, 110 Haemophilus influenzae and 50 Moraxella catarrhalis strains selected for their resistance phenotypes . Broth microdilution MICs and MBCs were determined according to NCCLS guidelines . RESULTS: The MIC90s were 0.25-0.5 mg/L for S . pneumoniae, including penicillin-, erythromycin-, levofloxacin- and multidrug-resistant strains . The MIC90s for Streptococcus pyogenes and Streptococcus agalactiae were 0.12 mg/L and for viridans streptococci, the MIC90 was 0.5 mg/L . Against S . aureus, including oxacillin- and levofloxacin-resistant strains, and vancomycin-intermediate strains, the MIC90 was 8 mg/L . Against beta-lactamase-negative and -positive H . influenzae, the MIC90s were 32 and 64 mg/L, respectively, and against both beta-lactamase-negative and -positive M . catarrhalis the MIC90 was 0.12 mg/L . In MBC studies, the ratio of MBC/MIC was 1:1 or 2:1 against 31% of S . pneumoniae, 33% of S . aureus, 63% of H . influenzae and 9% of M . catarrhalis . CONCLUSIONS: Although BB-83698 has reduced in vitro activity against H . influenzae, it is a potent antimicrobial with excellent activity against streptococci and Moraxella.

J Antimicrob Chemother, 2004 Apr, 53(4), 660 - 3 Epub 2004 Feb 18.
Disc diffusion susceptibility testing of Haemophilus influenzae by NCCLS methodology using low-strength ampicillin and co-amoxiclav discs; Karpanoja P et al.; OBJECTIVES: The objective of this multicentre study was to define the accuracy and reproducibility of the NCCLS disc diffusion method for Haemophilus influenzae against ampicillin and co-amoxiclav in Finnish clinical microbiology laboratories . Special attention was paid to the ability of the laboratories to detect beta-lactamase-negative ampicillin-resistant (BLNAR) strains . METHODS: Three BLNAR and two beta-lactamase-negative ampicillin-susceptible isolates (BLNAS)-originating from the American Type Culture Collection (ATCC) and UK National External Quality Assessment (UKNEQAS) schemes-were included in this study . Susceptibility tests for these isolates were performed in 26 clinical microbiology laboratories, in accordance with NCCLS guidelines . Additionally, low-strength discs for ampicillin (2 microg) and co-amoxiclav (3 microg) were tested . RESULTS: The low-strength discs for ampicillin and co-amoxiclav categorized more accurately BLNAR and BLNAS H . influenzae isolates than did the high-strength discs recommended by the NCCLS . In addition, the high-strength discs produced more major errors than the low-strength discs (22 versus six for ampicillin and 40 versus seven for co-amoxiclav) . Great variation occurred in the method regardless of the antibiotic concentration of the discs . CONCLUSIONS: The use of low-content ampicillin and co-amoxiclav discs is recommended for the susceptibility testing of H . influenzae . Interpretative criteria of S > or = 17 mm and R < or = 13 mm for both discs are suggested.

S Afr Med J, 2004 Jan, 94(1), 43 - 6
Safety and immunogenicity of two Haemophilus influenzae type b conjugate vaccines; Matjila MJ et al.; OBJECTIVES: Haemophilus influenzae type b (Hib) infection remains a major public health problem in the developing world . We evaluated the safety and immunogenicity of a new PRP-CRM197 conjugate Hib vaccine (Vaxem Hib, Chiron Vaccines), compared with the HibTITER vaccine (Wyeth-Lederle Vaccines), following the World Health Organisation (WHO)'s accelerated schedule which allows 4-week intervals between doses . STUDY DESIGN: A phase II, observer-blind, multicentre, randomised, controlled, non-inferiority study . METHODS: In total, 331 babies were immunised with either Vaxem Hib (N = 167) or HibTITER (N = 164) vaccine at 6, 10 and 14 weeks of age, in parallel with oral polio, diphtheriatetanus-pertussis and hepatitis B vaccines . Post-immunisation reactions were recorded after each immunisation and at follow-up visits . Anti-polyribosylribitol phosphate (PRP) antibodies were measured using enzyme-linked immunosorbent assays (ELISAs) before and 1 month after the third immunisation . RESULTS: Overall, there was no significant difference in the anti-PRP levels between the two groups . One month after the third immunisation, 76% of vaccinees in the Vaxem Hib group and 70% in the HibTITER group had anti-PRP antibody titres > or = 1.0 microgram/ml, while 96% of the Vaxem Hib group and 90% of the HibTITER group demonstrated anti-PRP antibody titres > or = 0.15 microgram/ml . The geometric mean titre at day 90 was 3.77 micrograms/ml for the Vaxem Hib and 3.0 micrograms/ml for the HibTITER groups . Although the Vaxem Hib vaccine produced more redness (6% versus 1%; p = 0.006) and swelling (5% versus 1%, p = 0.037), overall it was well tolerated compared with the HibTITER vaccine . There was no significant difference in vaccine-related elevated temperature (> or = 38 degrees C) between the two groups (p = 0.11) . CONCLUSION: Both vaccines showed comparable safety and immunogenicity profiles when administered to South African babies at 6, 10 and 14 weeks of age.

Infez Med, 1997, 5(2), 96 - 9
{Haemophilus influenzae type b in meningitis: antibiotic resistance in pediatric patients}; Ticca F et al.; A retrospective study on 357 children admitted to four Pediatric Infectious Disease Centers in Rome, affected by acute meningitis, during 10 years period, between January, 1, 1985 and December, 31, 1994 was carried out . Haemophilus influenzae type b was detected in 110 patients; all children aged between 1 month and 5 years; the maximum incidence (74.5%) was observed in patients under two years . The following diagnostic criteria were utilized: Gram stain of CSF; Latex test on CSF, blood, urine; CSF and blood cultures . The in vitro sensitivity of 65 isolates was tested by using the Kirby-Bauer method . We detected 15.3% of strains resistant to Ampicillin and 1.5% resistant to CAF . We also observed a high number of Hib strains resistant to Erythromycin and Cotrimoxazole . Only one strain Ceftriaxone resistant was isolated, confirming the high in vitro sensibility Hib to III generation cephalosporins that still remain the first choice drugs in Hib meningitis.

Br J Gen Pract, 2004 Jan, 54(498), 15 - 9
Pathogens involved in lower respiratory tract infections in general practice; Graffelman AW et al.; BACKGROUND: There are few investigations into the aetiology of lower respiratory tract infections (LRTIs) in general practice . AIM: To describe the aetiology of LRTI among adult patients in general practice in The Netherlands . DESIGN OF STUDY: Prospective observational study . SETTING: General practices in the Leiden region, The Netherlands . METHOD: Adult patients with a defined LRTI were included . Standard medical history and physical examination were performed . Sputum, blood and throat swabs were collected for diagnostic tests . Aetiological diagnosis, categorised as definite or possible, was based on the results of bacterial and viral cultures, serological techniques, and on polymerase chain reaction . Proportions of pathogens causing LRTI were assessed in relation to chest X-ray findings . RESULTS: A bacterial cause was established in 43 (30%), and a viral cause in 57 (39%) of the 145 patients with a LRTI . Influenza virus A was the most frequently diagnosed microorganism, followed by Haemophilus influenzae, and Mycoplasma pneumoniae . Streptococcus pneumoniae was found in 6% of the patients . CONCLUSIONS: Pathogens were found in two-thirds of the patients . In half of these patients there was a viral cause . Influenza virus A was the most frequently found pathogen . The treatment with antibiotics of at least one-third of the patients with LRTI was superfluous . This observation should result in changes in the prescription of antibiotics in LRTI.

Hist Cienc Saude Manguinhos, 2003, 10(Suppl 2), 697 - 724
{Vaccine innovations in Brazil: recent experiences and structural constraints}; Gadelha C et al.; This contribution to the current discussion on innovations in the area of science, technology, and health focuses on two experiences in Brazil's history: development of hepatitis B (HB) vaccines and of Hib vaccines against meningitis type B (Haemophilus influenzae), produced, respectively, at the Butanta Institute and at Bio-Manguinhos, which is the technical -scientific department of Fiocruz responsible for industrial production of immunobiologics . The analysis of these experiences offers a critical reference point for reflections both on scientific and technological development in the area of vaccines as well as on the role of leadership and institutional models relied upon to date . This analysis suggests that a new science and technology policy should be drawn up, one that would be capable of overcoming the barriers of dependence and of responding to the challenge of reinforcing local technological skills and know-how as a basic source of competitiveness and development in health.

Eur J Intern Med, 2003 Dec, 14(8), 488 - 492
Bronchiectasis in secondary care: a comprehensive profile of a neglected disease; Kelly MG et al.; Background: Bronchiectasis is poorly characterised in secondary care . Methods: Over 6 months, 410 bronchiectasis patients attended our clinics . One hundred randomly selected patients were characterised in detail . Results: Patients had a mean and standard error of mean (S.E.M.) age of 57 (2) years and a median and interquartile range (IQR) of three (two to four) reviews in the last 12 months . Aetiologies identified included tuberculosis (n=15), childhood pneumonia (n=7), fibrosis (n=6), connective tissue disease (n=6), whooping cough (n=5), childhood measles (n=4) and others (n=5) . There was widespread use of inhaled therapy . Treatments included oral antibiotics (n=77), corticosteroid courses (n=27) and intravenous antimicrobials (n=27, 12 domicillary) in the last year . Thirty patients had hospital admissions (13 because of the inability to administer domicillary antibiotics) . Haemophilus influenzae and Pseudomonas spp . were the commonest bacterial isolates . Patients culturing Pseudomonas spp . were older and had had more reviews and intravenous antibiotic courses . Conclusions: Bronchiectasis imposes a considerable burden on hospital services . Patients culturing Pseudomonas spp . impose a greater burden . Aetiology is often unknown . Therapies with unproven benefit are often used.

Rev Esp Quimioter, 2003 Dec, 16(4), 436 - 43
Activity of oral antibiotics against respiratory tract pathogens in Spain; Calvo A et al.; The aim of this study was to carry out a nationwide survey to assess the susceptibility of clinical isolates of four respiratory pathogens against nine antibiotics . Eight Spanish centers participated in the study, collecting a total of 977 isolates of Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae and Moraxella catarrhalis . The susceptibility of S . pneumoniae to penicillin was 37.46% susceptible, 30.43% intermediate and 32.11% resistant . MIC(90) of all antibiotics against this microorganism were 48 mg/l, except cefaclor, cefixime and azithromycin . For S . pyogenes, all the strains were susceptible to penicillins and cephalosporins, and azithromycin was the least active with a rate of resistance of 11.43% . A total of 95 isolates of H . influenzae were betalactamase positive (26.32%) . With regard to M . catarrhalis, only penicillin and amoxicillin showed MICs(90) >=8 mg/l.

J Trauma, 2004 Feb, 56(2), 296 - 301; discussion 301-3
A time-dependent analysis of intensive care unit pneumonia in trauma patients; Bochicchio GV et al.; BACKGROUND: Appropriate and timely antibiotic therapy to treat pneumonia in trauma patients is extremely important . We evaluated the incidence and microbiology of pneumonia stratified by days postadmission and risk factors . METHODS: Prospective data were collected on 714 trauma patients admitted to the intensive care unit over a 1-year period . Pneumonia was classified as community acquired (CAP) (< or = 3 days), early nosocomial (ENP) (4-6 days), or late nosocomial (LNP) (> or = 7 days) . In addition, pneumonia was classified as CAP only, nosocomial only (NI), or combination (CAP and NI, or ENP and LNP) pneumonia . Strict institutional guidelines were followed for diagnosis . RESULTS: One hundred eighty-two patients (25%) were diagnosed with 204 pneumonias over the study period . One hundred twenty-five (61%) of these pneumonias were ventilator associated . Staphylococcus aureus and Haemophilus influenzae were the most common pathogens isolated . Twenty-one percent of patients with CAP acquired an LNP (p < 0.025), in which Pseudomonas was the most common organism . Haemophilus caused LNP in 12% of patients . Cancer (p < 0.01), liver failure (p < 0.05), and age (p < 0.01) were predictive of nontypical pathogens in patients with CAP and ENP (p < 0.05) . Obesity was most predictive of increased ventilator days (p < 0.001) and intensive care unit length of stay (p < 0.001) . Increased age, alcohol abuse, and field airway were most predictive of mortality . CONCLUSION: Unanticipated pathogens were isolated in each class of pneumonia . The clinician must be aware of significant risk factors that may predispose patients to pathogens that are not ordinarily covered with standard antibiotic therapy.

Nucleic Acids Res, 2004 Feb 11, 32(3), 1050 - 8 Print 2004.
Biased distribution of DNA uptake sequences towards genome maintenance genes; Davidsen T et al.; Repeated sequence signatures are characteristic features of all genomic DNA . We have made a rigorous search for repeat genomic sequences in the human pathogens Neisseria meningitidis, Neisseria gonorrhoeae and Haemophilus influenzae and found that by far the most frequent 9-10mers residing within coding regions are the DNA uptake sequences (DUS) required for natural genetic transformation . More importantly, we found a significantly higher density of DUS within genes involved in DNA repair, recombination, restriction-modification and replication than in any other annotated gene group in these organisms . Pasteurella multocida also displayed high frequencies of a putative DUS identical to that previously identified in H.influenzae and with a skewed distribution towards genome maintenance genes, indicating that this bacterium might be transformation competent under certain conditions . These results imply that the high frequency of DUS in genome maintenance genes is conserved among phylogenetically divergent species and thus are of significant biological importance . Increased DUS density is expected to enhance DNA uptake and the over-representation of DUS in genome maintenance genes might reflect facilitated recovery of genome preserving functions . For example, transient and beneficial increase in genome instability can be allowed during pathogenesis simply through loss of antimutator genes, since these DUS-containing sequences will be preferentially recovered . Furthermore, uptake of such genes could provide a mechanism for facilitated recovery from DNA damage after genotoxic stress.

Pediatr Infect Dis J, 2004 Feb, 23(2), 127 - 31
Case-control studies of the effectiveness of vaccines: validity and assessment of potential bias; Shapiro ED; BACKGROUND: Because case-control studies of the effectiveness of vaccines are nonexperimental, it is difficult to assure that bias does not affect the validity of the results . METHODS: A case-control study of the effectiveness of vaccines against Haemophilus influenzae type b (Hib) was replicated with a "sham" study . Cases were children > or =18 months of age with invasive infection caused by either Hib (original study) or Streptococcus pneumoniae (sham study) between January 1988 and December 1990 . Controls were matched to the cases by both date and town of birth . RESULTS: Overall 34% of the 29 cases with invasive Hib infections and 64% of matched controls had received Hib vaccine . The effectiveness of Hib vaccines against infection with Hib was 88% (95% confidence interval, 57 to 97%; P < 0.01) . In the sham study 74% of the 62 cases with invasive pneumococcal infections and 74% of matched controls had received Hib vaccine . The effectiveness of Hib vaccines against pneumococcal infection was 0% (P = 0.9) . CONCLUSION: With the use of a virtually identical study design, vaccines against Hib were shown to be highly effective in preventing invasive Hib infections but were not effective in preventing invasive infections due to S . pneumoniae . Case-control studies are a valid method of assessing the effectiveness of vaccines.

Gesundheitswesen, 2004 Feb, 66 Suppl 1, S13 - 20
{Surveillance of notifiable infectious diseases in Bavaria - results in 2002}; Ludwig MS et al.; In 2002 nearly 36 000 cases of notifiable infectious diseases were reported in Bavaria representing a 10 % increase compared to 2001 (33 000 cases) . As in 2001, around 75 % of reported cases were gastrointestinal infections . Every third infection was due to salmonella . As compared to last year, the incidence of Norwalk-like virus infections increased fivefold . These infections occurred mostly as outbreaks in nursing homes, hospitals or other institutions, affecting as many as 200 persons . Other frequently reported infections in Bavaria are tuberculosis and hepatitis . The relatively high incidence of measles is mainly due to an outbreak in Coburg . The decline in the incidence of tuberculosis observed over the last years has ceased . Around 70 % of reported hepatitis cases were due to hepatitis C . It should be noted that these cases were a mixture of new infections and ongoing infections diagnosed for the first time . Of great epidemiological importance are diseases caused by meningitis pathogens . The incidence of meningococcal infections was practically unchanged as compared to last year . Around half of them were caused by serotype B, which is currently not preventable by vaccination . Meningitis caused by Haemophilus influenzae B is continually declining due to the high vaccination rate and is very rarely reported . Several cases of FSME were described . According to investigations carried out by health departments these infections were acquired in countries not yet classified as FSME risk areas . Hence, the endemics maps of FSME have to be revised . Two years of reporting according to IfSG (infectious disease control law) yielded very encouraging results, i . e . rapid accessibility of data, flexibility, complete and standardised reporting with high quality of data . We thank all the reporting physicians and laboratories and the staff of the Bavarian health departments for their continuous support.

J Immunother, 2004 Mar-Apr, 27(2), 124 - 35
Immunologic analysis of a phase I/II study of vaccination with MAGE-3 protein combined with the AS02B adjuvant in patients with MAGE-3-positive tumors; Vantomme V et al.; In a phase I/II study, patients with solid metastatic MAGE-3-positive tumors, mainly melanoma, were vaccinated with recombinant MAGE-3 protein combined with the immunologic adjuvant AS02B comprised of MPL and QS21 in an oil-in-water emulsion . The recombinant MAGE-3 protein was made up of a partial sequence of the protein D (ProtD) antigen of Haemophilus influenzae fused to the MAGE-3 sequence . The vaccine was given intramuscularly at 3-week intervals . Patients whose tumors stabilized or regressed after 4 vaccinations received 2 additional vaccinations at 6-week intervals . MAGE-3 and ProtD antibody and cellular immune responses were monitored after vaccination . Ninety-six percent (23/24) of the patients vaccinated with MAGE-3 protein in AS02B adjuvant elicited a significant anti-MAGE-3 IgG antibody response after 4 vaccinations, and all developed anti-ProtD IgG antibodies . For the detection of T-cell activity, total peripheral blood mononuclear cells were restimulated in vitro with MAGE-3- or ProtD-loaded autologous mature dendritic cells . In 30% of the evaluable patients vaccinated with the adjuvanted recombinant protein, IFNgamma production was increased in response to MAGE-3, and 2 patients (14% of evaluable patients) had a concomitant increase in IL-5 production . In 37% and 43% of the patients, respectively, IFNgamma or IL-5 production was increased in response to ProtD . It is concluded that vaccination of advanced cancer patients with MAGE-3 self-antigen in AS02B adjuvant is able to elicit MAGE-3-specific antibody and a T-cell response.

Pediatr Infect Dis J, 2004 Feb, 23(2 Suppl), S115 - 24
Management of recurrent and persistent acute otitis media: new options with familiar antibiotics; Arrieta A et al.; Recurrent and persistent acute otitis media (AOM) is a common problem, affecting close to 20% of children in their first years of life . It presents a therapeutic challenge to physicians, particularly as beta-lactamase-producing Haemophilus influenzae, as well as penicillin- and, more recently, macrolide-resistant Streptococcus pneumoniae are often responsible for AOM in these patients . Intramuscular ceftriaxone (50 mg/kg, once daily for 3 days) has been shown to be effective in treating nonresponsive AOM, but there have been relatively few clinical trials exploring oral antibiotic regimens . Higher doses of commonly used antibiotics (amoxicillin-clavulanate ratio, 14:1 ratio and azithromycin 60 mg/kg divided into three equal once daily doses) have demonstrated high rates of clinical success . These studies will be discussed together with a review of the clinical and microbiologic characteristics of this disease . We will also address the impact that the pneumococcal conjugate vaccine may have on the microbiology of recurrent and persistent AOM.

Rev Panam Salud Publica, 2003 Dec, 14(6), 377 - 84
Meningitis and pneumonia in Guatemalan children: the importance of Haemophilus influenzae type b and Streptococcus pneumoniae; Asturias EJ et al.; OBJECTIVE: To determine the epidemiology of Haemophilus influenzae type b (Hib) and Streptococcus pneumoniae invasive infections in hospitalized Guatemalan children . This is an important issue since Hib vaccine has not been incorporated into the routine immunization program in Guatemala and information from hospital records in 1995 indicated a low incidence of Hib and S . pneumoniae as causes of meningitis and invasive infections . METHODS: Children who were hospitalized in Guatemala City with clinical signs compatible with bacterial infections were evaluated for evidence of Hib or S . pneumoniae infection . Normally sterile body fluids were cultured, and antigen detection was performed on cerebrospinal fluid (CSF) and pleural fluid . RESULTS: Of 1 203 children 1-59 months of age hospitalized over a 28-month period, 725 of them (60.3%) had a primary diagnosis of pneumonia, 357 (29.7%) of meningitis, 60 (5.0%) of cellulitis, and 61 (5.1%) of sepsis and other conditions . Hib was identified in 20.0% of children with meningitis and S . pneumoniae in 12.9% . The average annual incidence of Hib meningitis was 13.8 cases per 100 000 children under 5 years of age, and 32.4% of meningitides caused by Hib and 58.7% of S . pneumoniae meningitides occurred prior to 6 months of age . Case fatality rates were 14.1%, 37.0%, and 18.0%, respectively, for children with Hib, S . pneumoniae, and culture-negative and antigen-negative meningitis . Prior antibiotic therapy was common and was associated with significant reductions in CSF-culture-positive results for children with other evidence of Hib or S . pneumoniae meningitis . CONCLUSIONS: Improvements in case detection, culture methods, and latex agglutination for antigen detection in CSF resulted in identification of Hib and S . pneumoniae as important causes of severe disease in Guatemalan children . Using a cutoff of > 10 white blood cells per cubic millimeter in CSF would improve the sensitivity for detection of bacterial meningitis and help estimate the burden of bacterial meningitis in Guatemala and other developing countries.

Nihon Kokyuki Gakkai Zasshi, 2004 Jan, 42(1), 68 - 74
{A retrospective analysis of community-acquired pneumonia between 2000 and 2002 in a community hospital}; Motomura K et al.; We previously reported a hospital-based retrospective study on community-acquired pneumonia (CAP) at Tagami Hospital, which was a community hospital, between 1994 and 1997 . This study was designed to clarify the etiology of CAP diagnosed between 2000 and 2002 . We analyzed a total of 124 cases of CAP in our hospital during the study period, and compared the results with the previous data . Identification of the causative organisms of CAP was based on gram staining, the morphology of the colonies, quantitative culture of the sputum, and the serological tests . During the study period, we determined the causative organisms in 42 cases (33.8%) . Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis were the major causative organisms . The severity of the cases was classified into three groups according to the guideline for CAP, which was edited by the Japanese Respiratory Society . The survival rates in the moderate and severe groups were significantly (p < 0.001) higher than that of the mild group, as analyzed by the Kaplan-Meier method, as follows: 70% (moderate) vs 100% (mild); and 40% (severe) vs 100% (mild) . In a total of 7 patients who died, we found the following risk factors: elderly male patients, bedridden status with cerebral infarction, and micro-aspiration, including recurrent pneumonia at short intervals of less than 17 days . Our study indicated that the JRS-edited guideline for CAP is a very useful tool for analyzing cases with CAP in Japan.

J Clin Microbiol, 2004 Feb, 42(2), 877 - 80
Characterization of Haemophilus segnis, an important cause of bacteremia, by 16S rRNA gene sequencing; Lau SK et al.; We describe the application of 16S rRNA gene sequencing in defining eight cases of bacteremia due to Haemophilus species other than Haemophilus influenzae (non-H . influenzae bacteremia) during a 7-year period . The first case of acute pyelonephritis due to Haemophilus segnis is also reported . In contrast to the extremely rare incidence of H . segnis infections reported previously, our results suggested that H . segnis is an important cause of non-H . influenzae bacteremia.

J Clin Microbiol, 2004 Feb, 42(2), 839 - 40
Development of a new serological test for serotyping Haemophilus parasuis isolates and determination of their prevalence in North America; Tadjine M et al.; Haemophilus parasuis causes polyserositis in swine . Fifteen serovars have been characterized by immunodiffusion test, but many field strains are not typeable . Isolates (n = 300) of H . parasuis from animals in North America were serotyped by a new indirect hemagglutination test . The test was rapid and effective for serotyping of H . parasuis, and serovars 4, 5, 13, and 7 were the most prevalent serotypes.

J Clin Microbiol, 2004 Feb, 42(2), 807 - 10
Emergence of nonencapsulated and encapsulated non-b-type invasive Haemophilus influenzae isolates in Portugal (1989-2001); Bajanca P et al.; Phenotypes and genetic relatedness of invasive Haemophilus influenzae strains were evaluated from 1989 through 2001 . Among 119 isolates, multidrug resistance decreased (from 50 to 0%), the level of H . influenzae serotype b (Hib) strains declined (from 81 to 16%), the level of noncapsulated strains rose (from 19 to 80%), and the first invasive H . influenzae serotype f strain was described . This study documents changes in invasive H . influenzae infections in Portugal, i.e., the emergence of non-type-b strains that are genetically diverse and unrelated to Hib.

J Clin Microbiol, 2004 Feb, 42(2), 524 - 9
Analysis of invasive Haemophilus influenzae infections after extensive vaccination against H . influenzae type b; Campos J et al.; Little clinical and microbiological information is available about invasive Haemophilus influenzae infection after widespread vaccination against H . influenzae type b (Hib) . We conducted an active community surveillance study on invasive H . influenzae during a 2-year period in a community of more than 5 million people after vaccination against Hib in children was introduced . The median incidence was 16.3 cases/100000 persons per year in children less than 1-year-old and 4.41 cases/100000 persons in children less than <5 years old . The highest incidence in adults was observed in patients greater than 70 years old . Clinical diagnoses included bacteremia, pneumonia, and meningitis . Of the H . influenzae-infected patients, 74.3% had underlying predisposing conditions, including impaired immunity and respiratory diseases . A total of 73.6% of the isolates were nontypeable and 16.5, 6.6, and 3.3% were types b, f, and e, respectively . Infections due to capsulated strains b, e, and f were evenly distributed between children and adults . Ampicillin and cotrimoxazole resistance occurred at frequencies of 24.2 and 48.4%, respectively . Antibiotic resistance was more prevalent in capsulated than in noncapsulated H . influenzae . Invasive isolates were highly resistant to antibiotics that were used infrequently in the community . Nontypeable H . influenzae were genetically much more heterogeneous than capsulated strains . Capsule-deficient mutants (b(-)) were not detected . Plasmid carriage was linked to antibiotic resistance and capsulated strains . Over the study period, the incidence of invasive H . influenzae infections, either encapsulated or not, did not increase . In the post-Hib vaccination era, most invasive infections were due to noncapsulated strains and occurred in the extreme ages of life in patients with predisposing conditions.

J Immunol, 2004 Feb 15, 172(4), 2431 - 8
Haemophilus influenzae type b-outer membrane protein complex glycoconjugate vaccine induces cytokine production by engaging human toll-like receptor 2 (TLR2) and requires the presence of TLR2 for optimal immunogenicity; Latz E et al.; Conjugate vaccines consisting of the capsular polysaccharide (PS) of Haemophilus influenzae type b (Hib) covalently linked to carrier proteins, unlike pure PS, are immunogenic in infants and have significantly reduced Hib infections in the United States, but require multiple doses to induce protective anti-PS Ab titers . Hib-meningococcal outer membrane protein complex (OMPC) conjugate vaccine, however, elicits protective anti-PS Ab titers after one dose . We found that OMPC and Hib-OMPC engaged human Toll-like receptor 2 (TLR2) expressed in human embryonic kidney (HEK) cells, inducing IL-8 production, and engaged mouse TLR2 on bone marrow-derived dendritic cells, inducing TNF release . Hib conjugated to the carrier proteins CRM(197) and tetanus toxoid did not engage TLR2 on HEK or dendritic cells . Engagement of TLR2 by Hib-OMPC was MyD88 dependent, as Hib-OMPC-induced TNF production was ablated in MyD88 knockout (KO) mice . Hib-OMPC was significantly less immunogenic in TLR2 KO mice, inducing lower Hib PS IgG and IgM titers compared with those in wild-type mice . Splenocytes from OMPC-immunized TLR2 KO mice also produced significantly less IL-6 and TNF-alpha than those from wild-type mice . Hib-OMPC is unique among glycoconjugate vaccines by engaging TLR2, and the ability of Hib-OMPC to elicit protective levels of Abs after one dose may be related to TLR2-mediated induction and regulation of cytokines produced by T cells and macrophages in addition to the peptide/MHC II-dependent recruitment of T cell help commonly afforded by carrier proteins . TLR2 engagement by an adjuvant or carrier protein may be a useful strategy for augmentation of the anti-PS Ab response induced by glycoconjugate vaccines.

Med Trop (Mars), 2003, 63(4-5), 481 - 5
{Purulent meningitis in children: special considerations for developing countries}; Reinert P; Bacterial meningitis is frequent in tropical zones as a result of recurring epidemics, meningococcal meningitis and near-total absence of anti-haemophilus type b vaccination . Recently the situation has been further complicated by the unexplained appearance of meningococcal W135 epidemics in Burkina-Faso . With regard to pneumococcal meningitis, the appearance of antibiotic strains is a worldwide phenomenon that has greatly increased the cost of antimicrobial treatment . Hopefully in the future children in tropical areas will be able to benefit from immunization using a conjugate vaccine combining anti-haemophilus b, meningococcal type a and W135 vaccination . In this regard the currently available 7 valence pneumococcal vaccine is poorly suited to tropical epidemiology and that an additional 4 serotypes would be needed to achieve 85% coverage.

Diagn Microbiol Infect Dis, 2004 Jan, 48(1), 55 - 7
Disk diffusion quality control guidelines for NVP-PDF 713: a novel peptide deformylase inhibitor; Anderegg TR et al.; NVP-PDF713 is a peptide deformylase inhibitor that has emerged as a candidate for treating Gram-positive infections and selected Gram-negative species that commonly cause community-acquired respiratory tract infections . This report summarizes the results of a multi-center (seven participants) disk diffusion quality control (QC) investigation for NVP PDF-713 using guidelines of the National Committee for Clinical Laboratory Standards and the standardized disk diffusion method . A total of 420 NVP-PDF 713 zone diameter values were generated for each QC organism . The proposed zone diameter ranges contained 97.6-99.8% of the reported participant results and were: Staphylococcus aureus ATCC 25923 (25-35 mm), Streptococcus pneumoniae ATCC 49619 (30-37 mm), and Haemophilus influenzae ATCC 49247 (24-32 mm) . These QC criteria for the disk diffusion method should be applied during the NVP-PDF 713 clinical trials to maximize test accuracy.

Clin Microbiol Infect, 2004 Apr, 10 Suppl 2, 18 - 27
Building in efficacy: developing solutions to combat drug-resistant S . pneumoniae; Jacobs MR; The development of our understanding of the pharmacokinetic (PK) and pharmacodynamic (PD) principles that determine antimicrobial efficacy has advanced substantially over the last 10 years . We are now in a position to use PK/PD principles to set targets for antimicrobial design and optimisation so that we can predict eradication of specific pathogens or resistant variants when agents are used clinically . Optimisation of PK/PD parameters to enable the treatment of resistant pathogens with oral agents may not be possible with many current agents, such as some cephalosporins, macrolides and fluoroquinolones . Aminopenicillins, however, such as amoxicillin, have linear PK and have a good safety profile even at high doses . The new pharmacokinetically enhanced oral formulation of amoxicillin/clavulanate, 2000/125 mg twice daily, was designed using PK/PD principles to be able to eradicate Streptococcus pneumoniae with amoxicillin MICs of up to and including 4 mg/L, which includes most penicillin-resistant isolates . For amoxicillin and amoxicillin/clavulanate, a time above MIC (T > MIC) of 35-40% of the dosing interval (based on blood levels) is predictive of high bacteriological efficacy . This target was met by the design of a unique bilayer tablet incorporating 437.5 mg of sustained-release sodium amoxicillin in one layer plus 562.5 mg of immediate-release amoxicillin trihydrate and 62.5 mg of clavulanate potassium in the second layer, with two tablets administered for each dose . This unique design extends the bacterial killing time by increasing the T > MIC to 49% of the dosing interval against pathogens with MICs of 4 mg/L, and 60% of the dosing interval against pathogens with MICs of 2 mg/L . Based on these results, this new amoxicillin/clavulanate formulation should be highly effective in treating respiratory tract infections due to drug-resistant S . pneumoniae as well as beta-lactamase-producing pathogens, such as Haemophilus influenzae and Moraxella catarrhalis.

Clin Microbiol Infect, 2004 Apr, 10 Suppl 2, 6 - 11
Elements of design: the knowledge on which we build; MacGowan AP; The time the free drug serum concentration of antibiotic remains above the pathogen MIC (T > MIC) determines bacteriological efficacy and emergence or selection of resistance for penicillin and amoxicillin with or without clavulanate . Multiple studies in animal and in-vitro models now support this conclusion . The size of the T > MIC (the pharmacokinetic/-dynamic target) is > 40-50% to maximise antibacterial effect and pathogen eradication for Streptococcus pneumoniae and probably also Haemophilus influenzae . The size of the T > MIC for optimal antibacterial effect is changed by host immune status but not by bacterial inoculum or mechanism of resistance . There is good animal evidence to support the prediction that, as long as the target T > MIC is achieved, strains of S . pneumoniae with amoxicillin MICs of 0.016 mg/L will respond to amoxicillin in the same way as those with MICs of 1-2 mg/L . Emergence of resistance to amoxicillin/clavulanate in S . pneumoniae is related to low T > MIC (< 20%) and also to the degree of population heterogeneity to amoxicillin . Selection of resistant strains of S . pneumoniae is also related to T > MIC . Monte Carlo simulations based on the pharmacokinetics of amoxicillin with or without clavulanate in humans are needed to best predict the likely efficacy of different amoxicillin dosing regimens . This approach adequately allows the considerable pharmacokinetic variability in amoxicillin handling by infected patients to be accounted for as well as differences in pathogen beta-lactam susceptibility.

Biopolymers, 2004 Feb 5, 73(2), 216 - 28
DNA melting profiles from a matrix method; Poland D; In this article we give a new method for the calculation of DNA melting profiles . Based on the matrix formulation of the DNA partition function, the method relies for its efficiency on the fact that the required matrices are very sparse, essentially reducing matrix multiplication to vector multiplication and thus making the computer time required to treat a DNA molecule containing N base pairs proportional to N(2) . A key ingredient in the method is the result that multiplication by the inverse matrix can also be reduced to vector multiplication . The task of calculating the melting profile for the entire genome is further reduced by treating regions of the molecule between helix-plateaus, thus breaking the molecule up into independent parts that can each be treated individually . The method is easily modified to incorporate changes in the assignment of statistical weights to the different structural features of DNA . We illustrate the method using the genome of Haemophilus influenzae .

Eur Radiol . 2004 Jan 29; {Epub ahead of print}
Community-acquired and nosocomial pneumonia; Herold CJ et al.; Pneumonia is one of the leading causes of morbidity, hospitalization, and mortality in both industrialized and developing countries . In particular, pulmonary infections acquired in the community, and pneumonias arising in the hospital setting, represent a major medical and economic problem and thus a continuous challenge to health care . For the radiologist, it is important to understand that community-acquired pneumonia (CAP) and nosocomial pneumonia (NP) share a number of characteristics, but should, in many respects be regarded as separate entities . CAP and NP arise in different populations, host different spectra of causative pathogens, and pose different challenges to both the clinician and the radiologist . CAP is generally seen in outpatients, is most frequently caused by Streptococcus pneumoniae, Mycoplasma pneumoniae, Haemophilus influenzae, and Chlamydia, and its radiologic diagnosis is relatively straightforward . NP, in contrast, develops in the hospital setting, is commonly caused by gram-negative bacteria, and may generate substantial problems for the radiologist . Overall, both for CAP and NP, imaging is an integral component of the diagnosis, important for classification and differential diagnosis, and helpful for follow-up.

Proteins, 2004 Feb 15, 54(3), 375 - 83
Solution structure of the highly acidic protein HI1450 from Haemophilus influenzae, a putative double-stranded DNA mimic; Parsons LM et al.; The solution structure of the acidic protein HI1450 from Haemophilus influenzae has been determined by NMR spectroscopy . HI1450 has homologues in ten other bacterial species including Escherichia coli, Vibrio cholerae, and Yersinia pestis but there are no functional assignments for any members of the family . Thirty-one of the amino acids in this 107-residue protein are aspartates or glutamates, contributing to an unusually low pI of 3.72 . The secondary structure elements are arranged in an alpha-alpha-beta-beta-beta-beta order with the two alpha helices packed against the same side of an anti-parallel four-stranded beta meander . Two large loops, one between beta1 and beta2 and the other between beta2 and beta3 bend almost perpendicularly across the beta-strands in opposite directions on the non-helical side of the beta-sheet to form a conserved hydrophobic cavity . The HI1450 structure has some similarities to the structure of the double-stranded DNA (dsDNA) mimic uracil DNA glycosylase inhibitor (Ugi) including the distribution of surface charges and the position of the hydrophobic cavity . Based on these similarities, as well as having a comparable molecular surface to dsDNA, we propose that HI1450 may function as a dsDNA mimic in order to inhibit or regulate an as yet unidentified dsDNA binding protein .

Acta Crystallogr D Biol Crystallogr, 2004 Feb, 60(Pt 2), 397 - 400 Epub 2004 Jan 23.
Refinement of Haemophilus influenzae diaminopimelic acid epimerase (DapF) at 1.75 A resolution suggests a mechanism for stereocontrol during catalysis; Lloyd AJ et al.; Diaminopimelate (DAP) epimerase (DapF) is central to the biosynthesis of both lysine and cell-wall peptidoglycan in many bacteria species . The peptidoglycan layer provides great potential for the development of novel antimicrobials as it is a uniquely prokaryotic feature . Crystals of recombinant Haemophilus influenzae DapF that diffract to beyond 2 A resolution have been obtained which facilitated the solution of the structure by molecular replacement at a resolution approximately 1 A higher than that previously determined . An analysis of the structure (i) in comparison to other PLP-independent racemaces and (ii) in relation to the catalytic mechanism and stereospecificity of DapF is presented.

Pediatr Infect Dis J, 2004 Jan, 23(1), 38 - 41
Outbreak of Haemophilus influenzae type b disease among fully vaccinated children in a day-care center; McVernon J et al.; BACKGROUND: Two cases of invasive Haemophilus influenzae type b (Hib) infection were reported in immunized children in a day nursery within 1 week . Both cases were younger than 18 months of age, cared for in the same room . We aimed to characterize carriage of Hib and response to eradication therapy in this setting . METHODS: Ninety-four children were enrolled in the nursery, cared for by 21 staff in 4 rooms, divided by age . Two children of a part time teacher also attended . Oropharyngeal swabs were taken to detect Hib carriage before introduction of rifampin prophylaxis (20 mg/kg/day for 4 days) . A questionnaire addressing risk factors for colonization was administered to parents and staff . Carriage was reassessed in children and carers from the same room as the index cases 1 month later . RESULTS: Eighty-nine children and all 21 staff were swabbed . Two additional Hib carriers, 1 child and 1 staff member, were identified from the same room as the cases . These isolates appeared identical with those causing invasive disease . Given the small numbers no clear risk factors for carriage could be confirmed . Compliance with rifampin prophylaxis was 97.4% . One month later no carriers were found among the 7 children and 3 staff tested from the room in which the cases were originally identified . CONCLUSIONS: Although immunization against Hib has resulted in a reduction in the incidence of this disease in the UK, individual protection cannot be assumed to be infallible . The importance of timely chemoprophylaxis of close contacts of a child with invasive Hib disease is reinforced.

Pediatr Infect Dis J, 2004 Jan, 23(1), 32 - 7
Are economic evaluations of vaccines useful to decision-makers? Case study of Haemophilus influenzae type b vaccines; Brinsmead R et al.; BACKGROUND: In concert with efforts to increase global provision, economic evaluations of newer and relatively costly vaccines have proliferated in the medical literature . The extent to which existing vaccine evaluations are useful to decision makers is not clear . We conducted a systematic review of published economic evaluations of conjugate Haemophilus influenzae type b (Hib) vaccine, anticipating that their usefulness to past and present decision makers would be limited by the quality of the analyses and by the extent to which the results were transferable to other settings . METHODS: We systematically identified economic evaluations of conjugate Hib vaccine . We appraised their quality according to a customized checklist and assessed the extent of and reasons for variability of the results . RESULTS: Quality assessment of the available economic evaluations disclosed a number of shortcomings, including the failure across all models to derive systematic estimates of vaccine efficacy as well as a lack of transparency in the costing of Hib disease treatment . Wide variations in results appeared to be caused primarily by epidemiologic and health system differences between settings and secondarily to methodologic differences between models . The generalizability of model results appeared low . CONCLUSIONS: There is scope for improving the overall quality of economic evaluations of Hib vaccination . Relevance to decision makers may also be increased by addressing local budget constraints and vaccine price . There is a need to better understand the decision process, particularly at the national level, to ensure the role of future economic evaluations as important decision tools in the implementation of new vaccines.

Infect Immun, 2004 Feb, 72(2), 1204 - 9
Haemophilus influenzae porin induces Toll-like receptor 2-mediated cytokine production in human monocytes and mouse macrophages; Galdiero M et al.; The production of proinflammatory cytokines is likely to play a major pathophysiological role in meningitis and other infections caused by Haemophilus influenzae type b (Hib) . Previous studies have shown that Hib porin contributes to signaling of the inflammatory cascade . We examined here the role of Toll-like receptors (TLRs) and the TLR-associated adaptor protein MyD88 in Hib porin-induced production of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) . Hib porin-induced TNF-alpha and IL-6 production was virtually eliminated in macrophages from TLR2- or MyD88-deficient mice . In contrast, macrophages from lipopolysaccharide (LPS)-hyporesponsive C3H/HeJ mice, which are defective in TLR4 function, responded normally to Hib porin . Moreover anti-TLR2 antibodies but not anti-TLR4 antibodies significantly reduced Hib porin-stimulated TNF-alpha and IL-6 release from the human monocytic cell line THP-1 . These data indicate that the TLR2/MyD88 pathway plays an essential role in Hib porin-mediated cytokine production . These findings may be useful in the development of alternative therapies aimed at reducing excessive inflammatory responses during Hib infections.

Infect Immun, 2004 Feb, 72(2), 1143 - 6
Haemophilus ducreyi strain ATCC 27722 contains a genetic element with homology to the vibrio RS1 element that can replicate as a plasmid and confer NAD independence on haemophilus influenzae; Munson RS Jr et al.; The nucleotide sequence of pNAD1, a plasmid from Haemophilus ducreyi identified on the basis of its ability to confer NAD independence on Actinobacillus pleuropneumoniae and H . influenzae, has been determined . In addition to containing the nadV gene, the plasmid contains homologues of the rstR and rstA genes, genes encoding repressor and replication proteins, respectively, in the Vibrio CTXphi and the Vibrio RS1 element, suggesting a single-stranded bacteriophage origin for pNAD1 . Tandem copies of the plasmid are integrated into the H . ducreyi 35000HP genome.

Antimicrob Agents Chemother, 2004 Feb, 48(2), 525 - 32
New class of bacterial phenylalanyl-tRNA synthetase inhibitors with high potency and broad-spectrum activity; Beyer D et al.; Phenylalanyl (Phe)-tRNA synthetase (Phe-RS) is an essential enzyme which catalyzes the transfer of phenylalanine to the Phe-specific transfer RNA (tRNA(Phe)), a key step in protein biosynthesis . Phenyl-thiazolylurea-sulfonamides were identified as a novel class of potent inhibitors of bacterial Phe-RS by high-throughput screening and chemical variation of the screening hit . The compounds inhibit Phe-RS of Escherichia coli, Haemophilus influenzae, Streptococcus pneumoniae, and Staphylococcus aureus, with 50% inhibitory concentrations in the nanomolar range . Enzyme kinetic measurements demonstrated that the compounds bind competitively with respect to the natural substrate Phe . All derivatives are highly selective for the bacterial Phe-RS versus the corresponding mammalian cytoplasmic and human mitochondrial enzymes . Phenyl-thiazolylurea-sulfonamides displayed good in vitro activity against Staphylococcus, Streptococcus, Haemophilus, and Moraxella strains, reaching MICs below 1 micro g/ml . The antibacterial activity was partly antagonized by increasing concentrations of Phe in the culture broth in accordance with the competitive binding mode . Further evidence that inhibition of tRNA(Phe) charging is the antibacterial principle of this compound class was obtained by proteome analysis of Bacillus subtilis . Here, the phenyl-thiazolylurea-sulfonamides induced a protein pattern indicative of the stringent response . In addition, an E . coli strain carrying a relA mutation and defective in stringent response was more susceptible than its isogenic relA(+) parent strain . In vivo efficacy was investigated in a murine S . aureus sepsis model and a S . pneumoniae sepsis model in rats . Treatment with the phenyl-thiazolylurea-sulfonamides reduced the bacterial titer in various organs by up to 3 log units, supporting the potential value of Phe-RS as a target in antibacterial therapy.

Vet Immunol Immunopathol, 2004 Feb, 97(3-4), 207 - 17
Reactive oxygen and nitrogen intermediates contribute to Haemophilus somnus lipooligosaccharide-mediated apoptosis of bovine endothelial cells; Sylte MJ et al.; Although Haemophilus somnus causes septicemia and vasculitis in cattle, relatively little is known about how H . somnus affects endothelial cells in vitro . We previously reported that H . somnus lipooligosaccharide (LOS)-induced activation of caspases-3, -8 and -9, and apoptosis of bovine pulmonary artery endothelial cells (BPAEC) in vitro . Previous reports indicate that the generation of reactive oxygen species (ROS) or reactive nitrogen intermediates (RNI) can contribute to the induction of apoptosis . In the present study, we sought to determine whether ROS and RNI are involved in LOS-mediated apoptosis of BPAEC . We found that H . somnus LOS induced the generation of ROS in BPAEC, which was blocked by pretreatment with membrane permeable ROS scavengers, such as dimethylsulfoxide (DMSO) and allopurinol (AP) . Addition of DMSO or AP significantly reduced H . somnus LOS-mediated caspase-3 activation . Addition of membrane impermeable ROS scavengers (e.g . catalase and superoxide dismutase), failed to block LOS-mediated caspase-3 activation, suggesting a role for intracellular generation of ROS in LOS-induced apoptosis of BPAEC . Addition of N(G)-nitro-L-arginine methyl ester (L-NAME) or aminoguanidine, which are selective inhibitors of nitric oxide synthase, blocked NO release and significantly reduced caspase-3 activation in LOS treated BPAEC . These data suggest H . somnus LOS triggers endogenous ROS and RNI production by endothelial cells, which contributes to apoptosis.

Med Pregl, 2003 Sep-Oct, 56(9-10), 457 - 9
{Suppurative middle ear infection as a complication after tympanostomy tube placement}; Baljosevic I et al.; INTRODUCTION: Suppurative otitis media after tympanostomy tube placement is the most frequent complication of this surgical intervention . Otorrhea that occurs in the first two weeks following tube placement is called early, late otorrhea occurs at least two weeks following placement . Early otorrhea is usually a result of either an infection that already existed when the tube was placed, or contamination of the external auditory canal during operation . Late otorrhea is mostly a result of upper respiratory tract infection . MATERIAL AND METHODS: Our investigation was performed at the ENT Department, Mother and Child Health Care Institute in Belgrade . The research included children treated for secretory or recurrent otitis media . RESULTS AND DISCUSSION: We have examined 411 children implanted with 796 tympanostomy tubes in the last three years . We investigated changes within two weeks after operation . Suppuration was recorded with 81 children (19.7%) . Staphylococcus aureus was established in 33 (40.7%) Pseudomonas aeruginosa in 26 (32%), Haemophilus influenzae in 12 (15%) and Streptococcus pneumoniae in 10 (12.3%) cases . All children were treated with antibiotic ear drops according to the antibiogram for a period of 7 days . Full recovery was achieved after treatment with Ciprofloxacin drops in 67%, Neomycin in 18% and Gentamycin in 9% of cases . In other cases a combination of drops and oral antibiotics was used . CONCLUSION: In cases of suppurative otitis media after implantation of tympanostomy tubes, the secret should be treated with suction and after that antibiotic drops should be applied during 5 to 7 days . If suppuration is persistent, drops should be used with oral antibiotics.

Microbes Infect, 2004 Jan, 6(1), 58 - 67
Characterization of two lipoproteins in Pasteurella multocida; Lo M et al.; An in vivo expression technology (IVET) system was previously developed and used to identify Pasteurella multocida genes, which are upregulated during infection of the host . Of the many genes identified, two encoded products which showed similarity to the Haemophilus influenzae lipoproteins, protein D and PCP, which have been shown to stimulate heterologous immunity against infection with H . influenzae . Therefore, the lipoprotein homologues in P . multocida, designated GlpQ and PCP, were investigated . GlpQ and PCP were shown to be lipoproteins by demonstrating that post-translational processing of the proteins was inhibited by globomycin . The P . multocida GlpQ homologue showed glycerophosphodiester phosphodiesterase enzyme activity, indicating that it is a functional homologue of other characterized GlpQ enzymes . Using surface immunoprecipitation, PCP was found to be surface exposed, but GlpQ was not . Non-lipidated forms of GlpQ and PCP were expressed and purified from Escherichia coli and used to vaccinate mice . However, mice were not protected from challenge with live P . multocida . The lipoproteins were then expressed in E . coli in the lipidated form and used to vaccinate mice and chickens . Protection against challenge with live P . multocida was not observed.

Pediatr Neurosurg, 2003 Dec, 39(6), 285 - 90
Reflections on shunt infection; Kanev PM et al.; The placement and revision of ventriculoperitoneal (VP) shunts remains a mainstay in the surgical treatment of hydrocephalus . While the North American infection rate averages nearly 8-10%, published infection rates for VP shunt infection below 1% have been reported . We retrospectively reviewed shunt operations by a single surgeon over 62 months to analyze the infection rate . In 62 months, we performed 526 shunt placements or revisions in patients up to 18 years of age . There were 7 shunt infections (1.33%) . In 5 cases, the organism was Staphylococcus epidermidis, and a single shunt each was infected with Haemophilus influenzae and Staphylococcus aureus . Each infection was treated with external ventriculostomy drainage and intravenous antibiotics . The new shunt was placed at a new incision site after at least 5 days of sterile spinal fluid cultures . The mean follow-up among these patients after shunt insertion was 25 months . VP shunting remains the most common operation for hydrocephalus . Infections are linked with seizures, higher future risks of shunt infection and malfunction, and reduced IQ and school performance . Our infection rate during 62 months was limited to 1.33% . Uniform surgical technique, limited hardware and skin edge manipulation and double gloving may be important factors in limiting shunt infections .

Int J Antimicrob Agents, 2004 Jan, 23(1), 44 - 51
PROTEKT 1999-2000: a multicentre study of the antibiotic susceptibility of respiratory tract pathogens in Hong Kong, Japan and South Korea; Inoue M et al.; A multicentre surveillance study performed in the Far East during 1999-2000 investigated the in vitro activity of >20 antibacterials against common respiratory pathogens . In Hong Kong, Japan, and South Korea, 57.1, 44.5 and 71.5% Streptococcus pneumoniae were penicillin-resistant and 71.4, 77.9 and 87.6% were erythromycin-resistant, respectively . Overall, >90% of penicillin-resistant strains were also macrolide-resistant . All strains were susceptible to telithromycin . Fluoroquinolone-resistant isolates in Japan (1.3%), Hong Kong (14.3%) and South Korea (2.9%) were mostly co-resistant to penicillin, macrolides and tetracycline . Beta-lactamase production by Haemophilus influenzae isolates was 8.5% in Japan, 17.1% in Hong Kong and 64.7% in strains from South Korea . A single (0.27%) BLNAR isolate was obtained in Japan . There was no fluoroquinolone resistance . Moraxella catarrhalis was inhibited by telithromycin at <or=0.5mg/l and remained susceptible to macrolides, fluoroquinolones and amoxicillin-clavulanate . Resistance to antibacterials, particularly penicillin and macrolides, has reached high but stable levels in the Far East and the presence of multiply-resistant pneumococci is well established.

Int J Antimicrob Agents, 2004 Jan, 23(1), 39 - 43
Antimicrobial resistance of Streptococcus pneumoniae and Haemophilus influenzae isolated from children with community-acquired respiratory tract infections in Central Poland; Semczuk K et al.; Resistance to commonly used antimicrobial agents among the key respiratory pathogens is increasing worldwide and therefore a rational choice of an empirical treatment requires knowledge of both global and local resistance patterns . The susceptibility of 185 Streptococcus pneumoniae and 169 Haemophilus influenzae isolates collected from January 1999 to May 2002 at the Children's Memorial Health Institute, Warsaw, Poland, from 351 children with community-acquired respiratory tract infections (RTIs) has been determined . Of S . pneumoniae isolates, 84% were susceptible to penicillin, 91% to cefaclor, 95% to cefuroxime, 98% to cefotaxime, 79% to erythromycin, 46% to co-trimoxazole, 82% to clindamycin and 59% to tetracycline . The majority (83%) of erythromycin-resistant isolates tested carried the erm(B) gene, conferring the MLS(B) phenotype . All tetracycline-resistant S . pneumoniae strains analysed were tet(M) positive and tet(O) negative . A total of 24% of H . influenzae isolates were beta-lactamase-positive . H . influenzae susceptibility to amoxicillin/clavulanate, cefaclor, cefuroxime, azithromycin, tetracycline and co-trimoxazole was 100, 89, 94, 96, 96 and 43%, respectively.

Int J Antimicrob Agents, 2004 Jan, 23(1), 25 - 31
In vivo activity of amoxicillin/clavulanic acid and erythromycin in experimental otitis media caused by Streptococcus pneumoniae plus Haemophilus influenzae; Parra A et al.; A gerbil model of acute otitis media induced by Streptococcus pneumoniae plus Haemophilus influenzae was used to assess the efficacy of amoxicillin/clavulanic acid (A/C) (1.5/0.3, 2.5/0.5 and 10/2 mg/kg) and erythromycin (2.5, 10, 20 and 50 mg/kg) with or without acetaminophen . The amoxicillin/clavulanic acid MIC was 1/0.5 mg/l for both organisms and the erythromycin MICs were 0.12 and 4 mg/l for S . pneumoniae and H . influenzae, respectively . The organisms were inoculated directly into the middle ear (ME) and antibiotic treatment started 2 h post-inoculation and continued at 8h intervals for three doses . Acetaminophen was administered at 50 mg/kg . Samples for bacterial counting were obtained from the ME on day 2 . Amoxicillin/clavulanic acid peri-MIC concentrations in ME were effective in eradicating both organisms . Despite the inflammation induced by S . pneumoniae, erythromycin did not eradicate H . influenzae at ME concentrations (2.4 mg/l for erythromycin 50 mg/kg) higher than those obtained in humans but lower than the MIC.

Proteomics, 2004 Jan, 4(1), 46 - 58
Comparing expression level-dependent features in codon usage with protein abundance: an analysis of 'predictive proteomics'; McHardy AC et al.; Synonymous codon usage is a commonly used means for estimating gene expression levels of Escherichia coli genes and has also been used for predicting highly expressed genes for a number of prokaryotic genomes . By comparison of expression level-dependent features in codon usage with protein abundance data from two proteome studies of exponentially growing E . coli and Bacillus subtilis cells, we try to evaluate whether the implicit assumption of this approach can be confirmed with experimental data . Log-odds ratio scores are used to model differences in codon usage between highly expressed genes and genomic average . Using these, the strength and significance of expression level-dependent features in codon usage were determined for the genes of the Escherichia coli, Bacillus subtilis and Haemophilus influenzae genomes . The comparison of codon usage features with protein abundance data confirmed a relationship between these to be present, although exceptions to this, possibly related to functional context, were found . For species with expression level-dependent features in their codon usage, the applied methodology could be used to improve in silico simulations of the outcome of two-dimensional gel electrophoretic experiments.

J Med Microbiol, 2004 Feb, 53(Pt 2), 115 - 7
Automation of a fluorescence-based multiplex PCR for the laboratory confirmation of common bacterial pathogens; Smith K et al.; A fluorescence-based multiplex PCR was automated for the simultaneous detection of Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae in clinical samples from patients with suspected meningitis . Sensitivity of one to two genome copies per 100 microl sample and specificity of 100% for each organism were shown . Automation of DNA extraction, liquid handling, PCR and analysis are achieved on a single platform, which enables a high throughput and rapid turnaround of clinical samples that, in turn, leads to faster diagnosis . This is ultimately beneficial to the treatment of the patient and for public health management.

J Antimicrob Chemother, 2004 Feb, 53(2), 399 - 402 Epub 2004 Jan 16.
Antimicrobial resistance in the nasopharyngeal flora of children with acute maxillary sinusitis and maxillary sinusitis recurring after amoxicillin therapy; Brook I et al.; OBJECTIVE: To investigate the antimicrobial susceptibility of the organisms isolated from the nasopharynx of children who present with acute maxillary sinusitis (AMS) or maxillary sinusitis that recurred (RMS) after amoxicillin therapy . METHODS: Analysis of nasopharyngeal cultures obtained from 70 patients, 42 with AMS and 28 with RMS . RESULTS: Thirty-eight potentially pathogenic organisms were recovered in 36 (86%) of the children from the AMS group, and 40 were isolated from 26 (93%) of the children from the RMS group . The organisms isolated were Streptococcus pneumoniae (21 isolates), Haemophilus influenzae non-type b (17), Moraxella catarrhalis (15), Streptococcus pyogenes (13) and Staphylococcus aureus (12) . Resistance to the eight antimicrobial agents used was found in 34 instances in the AMS group compared to 93 instances in the RMS group (P < 0.005) . The difference between AMS and RMS was significant with S . pneumoniae resistance to amoxicillin (P < 0.0025), to co-amoxiclav (P < 0.0025), to trimethoprim-sulfamethoxazole (P < 0.05), to cefixime (P < 0.05), and to azithromycin (P < 0.05), and for H . influenzae to amoxicillin (P < 0.025) . CONCLUSIONS: These data illustrate the higher recovery rate of antimicrobial-resistant S . pneumoniae and H . influenzae from the nasopharynx of children who had maxillary sinusitis that recurred after amoxicillin therapy than those with AMS.

J Antimicrob Chemother, 2004 Feb, 53(2), 144 - 8 Epub 2004 Jan 16.
Activity of gemifloxacin against Streptococcus pneumoniae and Haemophilus influenzae; Morrissey I et al.; This review focuses on the activity of gemifloxacin, a new respiratory fluoroquinolone, against the two most important bacterial pathogens associated with lower respiratory tract infections, namely Streptococcus pneumoniae and Haemophilus influenzae.

Nihon Kokyuki Gakkai Zasshi, 2003 Dec, 41(12), 868 - 73
{Clinical features of Q fever pneumonia}; Okimoto N et al.; We report four cases of Q fever pneumonia diagnosed using PanBio Coxilla burnetii ELISA . The patients, a 21-year-old woman, a 53-year-old man, a 74-year-old man and a 87-year-old man, were among 284 with community-acquired pneumonia who were treated as inpatients from March 2001 till March 2003 . The frequency of Q fever pneumonia in community-acquired pneumonia was 1.4% . The 21-year-old woman was a typical case of Q fever pneumonia, since her clinical features showed 1 . the breeding of cats, 2 . development from a fever and non-productive caught in March, 3 . multiple soft consolidations in the chest radiograph, 4 . normal WBC count, 5 . cure by administration of clarithromycin . The pneumonias of the other 3 cases were considered to be mixed infections, with bacteria such as Streptococcus pneumoniae and Haemophilus influenzae . Their clinical features were 1 . elderly male patients with underlying diseases, 2 . development from fever and cough with purulent sputum in winter, 3 . coarse crackle on auscultation, 4 . consolidation with pleural effusion in chest radiograph, 5 . leukocytosis, elevation of BUN, hyponatremia, 6 . a few cases with unfavorable prognoses despite medication with carbapenem and minocycline . These findings suggested that two types of pneumonia exist; one with the usual features of atypical pneumonia, and the other presenting the clinical features of bacterial pneumonia of the elderly due to a mixed infection including C . burnetti.

J Biol Chem, 2004 Apr 9, 279(15), 14679 - 85 Epub 2004 Jan 15.
The Haemophilus influenzae Hia autotransporter contains an unusually short trimeric translocator domain; Surana NK et al.; Gram-negative bacterial autotransporter proteins are a growing group of virulence factors that are characterized by their ability to cross the outer membrane without the help of accessory proteins . A conserved C-terminal beta-domain is critical for targeting of autotransporters to the outer membrane and for translocation of the N-terminal "passenger" domain to the bacterial surface . We have demonstrated previously that the Haemophilus influenzae Hia adhesin belongs to the autotransporter family, with translocator activity residing in the C-terminal 319 residues . To gain further insight into the mechanism of autotransporter protein translocation, we performed a structure-function analysis on Hia . In initial experiments, we generated a series of in-frame deletions and a set of chimeric proteins containing varying regions of the Hia C terminus fused to a heterologous passenger domain and discovered that the final 76 residues of Hia are both necessary and sufficient for translocation . Analysis by flow cytometry revealed that the region N-terminal to this shortened translocator domain is surface localized, further suggesting that this region is not involved in beta-barrel formation or in translocation of the passenger domain . Western analysis demonstrated that the translocation-competent regions of the C terminus migrated at masses consistent with trimers, suggesting that the Hia C terminus oligomerizes . Furthermore, fusion proteins containing a heterologous passenger domain demonstrated that similarly small C-terminal regions of Yersinia sp . YadA and Neisseria meningitidis NhhA are translocation-competent . These data provide experimental support for a unique subclass of autotransporters characterized by a short trimeric translocator domain.

J Antimicrob Chemother, 2004 Jan, 53 Suppl 1, i3 - 20
Augmentin (amoxicillin/clavulanate) in the treatment of community-acquired respiratory tract infection: a review of the continuing development of an innovative antimicrobial agent; White AR et al.; Amoxicillin/clavulanate (Augmentin) is a broad-spectrum antibacterial that has been available for clinical use in a wide range of indications for over 20 years and is now used primarily in the treatment of community-acquired respiratory tract infections . Amoxicillin/clavulanate was developed to provide a potent broad spectrum of antibacterial activity, coverage of beta-lactamase-producing pathogens and a favourable pharmacokinetic/pharmacodynamic (PK/PD) profile . These factors have contributed to the high bacteriological and clinical efficacy of amoxicillin/clavulanate in respiratory tract infection over more than 20 years . This is against a background of increasing prevalence of antimicrobial resistance, notably the continued spread of beta-lactamase-mediated resistance in Haemophilus influenzae and Moraxella catarrhalis, and penicillin, macrolide and quinolone resistance in Streptococcus pneumoniae . The low propensity of amoxicillin/clavulanate to select resistance mutations as well as a favourable PK/PD profile predictive of high bacteriological efficacy may account for the longevity of this combination in clinical use . However, in certain defined geographical areas, the emergence of S . pneumoniae strains with elevated penicillin MICs has been observed . In order to meet the need to treat drug-resistant S . pneumoniae, two new high-dose amoxicillin/clavulanate formulations have been developed . A pharmacokinetically enhanced tablet dosage form of amoxicillin/clavulanate 2000/125 mg twice daily (available as Augmentin XR in the USA), has been developed for use in adult respiratory tract infection due to drug-resistant pathogens, such as S . pneumoniae with reduced susceptibility to penicillin, as well as beta-lactamase-producing H . influenzae and M . catarrhalis . Amoxicillin/clavulanate 90/6.4 mg/kg/day in two divided doses (Augmentin ES-600) is for paediatric use in persistent or recurrent acute otitis media where there are risk factors for the involvement of beta-lactamase-producing strains or S . pneumoniae with reduced penicillin susceptibility . In addition to high efficacy, amoxicillin/clavulanate has a well known safety and tolerance profile of the two new high-dose formulations are not significantly different from those of conventional formulations . Amoxicillin/clavulanate is included in guidelines and recommendations for the treatment of bacterial sinusitis, acute otitis media, community-acquired pneumonia and acute exacerbations of chronic bronchitis . Amoxicillin/clavulanate continues to be an important agent in the treatment of community-acquired respiratory tract infections, both now and in the future.

Scand J Infect Dis, 2003, 35(11-12), 851 - 7
Impact of conventional chemotherapy on levels of antibodies against vaccine-preventable diseases in children treated for cancer; Reinhardt D et al.; Intensive chemotherapy in children with malignancies causes partial immune deficiency, including long-term impairment of humoral immunity . We investigated the levels of antibodies against measles, mumps, polio, rubella, diphtheria, tetanus, and Haemophilus type b (Hib) in 139 children at the time of diagnosis of the malignant disease, during chemotherapy, after cessation of intensive treatment, and after re-vaccination . In general, cytostatic therapy resulted in a significant lowering of antibody levels . A decline of antibodies below the protective level as a consequence of cytostatic treatment was observed in 6% of the children for measles and mumps, in 18%, 12%, and 25% for polio types 1, 2, and 3, and in 21% for diphtheria . By contrast, rubella and tetanus antibodies remained within the protective range in all cases of this study . Re-vaccination 3 to 5 months after cessation of chemotherapy produced antibody levels about as high as those measured prior to therapy . Only 6 out of 83 children with previously positive antigen titres did not respond to re-vaccination . Vaccination or re-vaccination failed in 5 of 13 non-responders for more than 1 antigen, indicating a decreased reactability to vaccinations in some patients.

Lancet Infect Dis, 2004 Jan, 4(1), 40 - 3
T helper cells and efficacy of Haemophilus influenzae type b conjugate vaccination; McVernon J et al.; A small number of fully vaccinated children in the UK have experienced invasive Haemophilus influenzae type b (Hib) infection . A rise in disease in recent years has been associated with lower vaccine-induced antibody levels over the first 5 years of life, forcing greater dependence on immunological memory for protection . This has necessitated the introduction of a catch-up campaign, designed to boost immunity in children aged 6 months to 4 years of age . We suggest that the conjugate vaccine's inability to induce pathogen specific helper T cells, combined with a loss of natural boosting due to reduced circulation of Hib, may have contributed to the rising incidence of invasive disease 10 years after introduction of the conjugate vaccine . If so, the changing epidemiology of Hib infection in the UK may in part reflect the failure of a subunit vaccine to activate adequately all the necessary components of the immune system . This observation has implications for optimal scheduling of more recently licensed meningococcal and pneumococcal conjugate vaccines.

Am J Respir Med, 2003, 2(1), 39 - 54
Optimizing treatment outcomes in severe community-acquired pneumonia; de Castro FR et al.; Severe community-acquired pneumonia (CAP) is a life-threatening condition that requires intensive care unit (ICU) admission . Clinical presentation is characterized by the presence of respiratory failure, severe sepsis, or septic shock . Severe CAP accounts for approximately 5-35% of hospital-treated cases of pneumonia with the majority of patients having underlying comorbidities . The most common pathogens associated with this disease are Streptococcus pneumoniae, Legionella spp., Haemophilus influenzae, and Gram-negative enteric rods . Microbial investigation is probably helpful in the individual case but is likely to be more useful for defining local antimicrobial policies . The early and rapid initiation of empiric antimicrobial treatment is critical for a favorable outcome . It should include intravenous beta-lactam along with either a macrolide or a fluoroquinolone . Modifications of this basic regimen should be considered in the presence of distinct comorbid conditions and risk factors for specific pathogens . Other promising nonantimicrobial new therapies are currently being investigated . The assessment of severity of CAP helps physicians to identify patients who could be managed safely in an ambulatory setting . It may also play a crucial role in decisions about length of hospital stay and time of switching to oral antimicrobial therapy in different groups at risk . The most important adverse prognostic factors include advancing age, male sex, poor health of patient, acute respiratory failure, severe sepsis, septic shock, progressive radiographic course, bacteremia, signs of disease progression within the first 48-72 hours, and the presence of several different pathogens such as S . pneumoniae, Staphylococcus aureus, Gram-negative enteric bacilli, or Pseudomonas aeruginosa . However, some important topics of severity assessment remain controversial, including the definition of severe CAP . Prediction rules for complications or death from CAP, although far from perfect, should identify the majority of patients with severe CAP and be used to support decision-making by the physician . They may also contribute to the evaluation of processes and outcomes of care for patients with CAP.

Am J Respir Med, 2003, 2(3), 221 - 33
Treatment of community-acquired lower respiratory tract infections during pregnancy; Lim WS et al.; The incidence of lower respiratory tract infection (LRTI) in women of child-bearing age is approximately 64 per 1000 population . The spectrum of illness ranges from acute bronchitis, which is very common, through influenza virus infection and exacerbations of underlying lung disease, to pneumonia, which, fortunately is uncommon (<1.5% LRTI), but can be severe . Acute bronchitis is generally mild, self-limiting and usually does not require antibacterial therapy . Influenza virus infection in pregnant women has been recently related to increased hospitalization for acute cardiorespiratory conditions . At present, the safety of the newer neuraminidase inhibitors for the treatment of influenza virus infection has not been established in pregnancy and they are not routinely recommended . In influenza virus infection complicated by pneumonia, antibacterial agents active against Staphylococcus aureus and Streptococcus pneumoniae superinfection should be used.There are few data on infective complications of asthma or COPD in pregnancy . The latter is rare, as patients with COPD are usually male and aged over 45 years . Management is the same as for nonpregnant patients.The incidence and mortality of pneumonia in pregnancy is similar to that in nonpregnant patients . Infants born to pregnant patients with pneumonia have been found to be born earlier and weigh less than controls . Risk factors for the development of pneumonia include anemia, asthma and use of antepartum corticosteroids and tocolytic agents . Based on the few available studies, the main pathogens causing pneumonia are S . pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae and viruses . Beta-Lactam and macrolide antibiotics therefore remain the antibiotics of choice in terms of both pathogen coverage and safety in pregnancy . In HIV-infected pregnant patients, recurrent bacterial pneumonia, but not Pneumocystis carinii pneumonia (PCP), is more common than in nonpregnant patients . Trimethoprim/sulfamethoxazole (cotrimoxazole) has not definitely been associated with adverse clinical outcomes despite theoretical risks . Currently it is still the treatment of choice in PCP, where mortality remains high.In conclusion, there are few data specifically related to pregnant women with different types of LRTI . Where data are available, no significant differences compared with nonpregnant patients have been identified . In considering the use of any therapeutic agent or investigation in pregnant patients with LRTI, safety aspects must be carefully weighed against potential benefit . Otherwise, management strategies should not differ from those for nonpregnant patients . Further research in this area is warranted.

Arch Ophthalmol, 2004 Jan, 122(1), 65 - 9
Treatment of experimental bacterial keratitis with topical trovafloxacin; Barequet IS et al.; OBJECTIVE: To investigate the therapeutic role of trovafloxacin mesylate, a newer-generation fluoroquinolone with an expanded spectrum of activity, in the treatment of experimental bacterial keratitis . METHODS: Susceptibility studies were performed on various strains of ocular isolates to determine the minimum inhibitory concentration (MIC) of trovafloxacin compared with ciprofloxacin and ofloxacin, using the E-test method . Pharmacokinetic studies were performed by a single topical administration of trovafloxacin to rabbit eyes with either an intact or denuded corneal epithelium . Aqueous humor, vitreous, and corneal concentrations of trovafloxacin were determined at different time points . Experimental bacterial keratitis studies were performed in rabbit eyes . Three identical studies were conducted using Staphylococcus aureus, Streptococcus pneumoniae, or Pseudomonas aeruginosa . Therapy groups included 0.5% trovafloxacin, 0.3% ciprofloxacin, 0.3% ofloxacin, and isotonic sodium chloride solution . After 12 hours of drops administration, corneas were excised, homogenized, and serially plated . The main outcome measure was quantitative bacteriologic analysis for residual colony-forming units . RESULTS: In vitro susceptibility study findings indicated that the MIC of trovafloxacin was significantly lower than the MIC of ciprofloxacin and ofloxacin for S . aureus, S . pneumoniae, and Haemophilus influenzae, lower than the MIC of ciprofloxacin and ofloxacin for Staphylococcus epidermidis, and intermediate between ciprofloxacin and ofloxacin for P . aeruginosa . Pharmacokinetic studies showed a significant concentration of trovafloxacin in the treated corneas, especially in eyes with a denuded epithelium . All serum samples had undetectable trovafloxacin concentrations . Experimental keratitis studies showed a statistically significant decrease of colony-forming units in trovafloxacin-treated eyes in the S . aureus model and a similar decrease in the S pneumoniae and P aeruginosa models . CONCLUSIONS: Topical 0.5% trovafloxacin proved to be an effective ocular medication for the therapy of gram-positive and gram-negative keratitis . Clinical Relevance Trovafloxacin may provide an excellent therapeutic alternative in bacterial keratitis.

Emerg Infect Dis, 2003 Nov, 9(11), 1486 - 8
Polymerase chain reaction assay and bacterial meningitis surveillance in remote areas, Niger; Sidikou F et al.; To compensate for the lack of laboratories in remote areas, the national reference laboratory for meningitis in Niger used polymerase chain reaction (PCR) to enhance the surveillance of meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae . PCR effectively documented the wide geographic spread of N . meningitidis serogroup W135.

Emerg Infect Dis, 2003 Nov, 9(11), 1475 - 8
Levofloxacin treatment failure in Haemophilus influenzae pneumonia; Bastida T et al.; We describe the first case of failure of oral levofloxacin treatment of community-acquired pneumonia caused by Haemophilus influenzae . The strain showed cross-resistance to fluoroquinolones and carried four mutations in quinolone resistance-determining regions of DNA gyrase and topoisomerase IV genes.

Bull Soc Pathol Exot, 2003 Nov, 96(4), 313 - 6
{Agents of community acquired purulent meningitis in the child: epidemiologic trends in Abidjan, Côte d'Ivoire, from the year 1995 to 2000}; Faye-Kette H et al.; OBJECTIVE: To assess prevalence and trends of community acquired bacterial meningitis in childhood in a tertiary-care hospital before introduction of the HIB conjugate vaccine . STUDY DESIGN: Laboratory based data were recorded from January 1995 to December 2000 on two hundred and eighty seven children with bacterial meningitis . Identification of bacterial agents was performed with conventional methods . Information including age, gender, bacterial aetiology of meningitis, month and annual prevalence of agents was examined . RESULTS: The age of infected children ranges from 1 to 10 years with an average and median age of 34.2 months and 12 months respectively . Fifty five percent of children were male . The overall prevalence of agents were respectively 47.8% for Streptococcus pneumoniae followed by Haemophilus influenzae 39% and Neisseria meningitidis 13.2% with predominance of serogroup C . Stratification by age group shows that Haemophilus influenzae was the most common agent among children < 1 year of age following by S . pneumoniae and N . meningitidis . After 5 years, the number of cases of S . pneumoniae and N . meningitidis was prevalent . After 10 years, N . meningitidis was the first aetiology of bacterial meningitis . The six years data recorded highlighted the high and stable prevalence of H . influenzae B and S . pneumoniae and the low prevalence of N . meningitidis and high incidence of invasive meningococcal, pneumococcal and Haemophilus influenzae during the six years between September and February . CONCLUSION: Conjugated HIB vaccine is needed in our country to lower incidence of H . influenzae meningitis as already seen in developed countries . Continuous surveillance is necessary to monitor the disease trends, serotype distribution and antimicrobial susceptibility in order to implement appropriate public health interventions against community acquired bacterial meningitis.

J Clin Microbiol, 2004 Jan, 42(1), 412 - 4
Survival of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis frozen in skim milk- tryptone-glucose-glycerol medium; Kaijalainen T et al.; In STGG (skim milk, tryptone, glucose, glycerol) medium at -80 degrees C, Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis isolates survived for at least 3 years, and the same species have survived in nasopharyngeal swabs for at least 1.5 years . At -20 degrees C, S . pneumoniae and M . catarrhalis survived for 1.5 years, but H . influenzae survived for only 2 months.

J Clin Microbiol, 2004 Jan, 42(1), 362 - 5
Possible high rate of transmission of nontypeable Haemophilus influenzae, including beta-lactamase-negative ampicillin-resistant strains, between children and their parents; Watanabe H et al.; The possible transmission of nontypeable Haemophilus influenzae between children and their parents was evaluated in 18 pairs of subjects from 15 families . Of the 33 isolates, 31 were found to be beta-lactamase negative, including 10 beta-lactamase-negative, ampicillin (AMP)-resistant (BLNAR) strains (AMP MIC, >or=1.0 microg/ml) and 2 were beta-lactamase producing . Molecular typing by pulsed-field gel electrophoresis (PFGE) showed that 10 BLNAR isolates had 6 patterns, 23 non-BLNAR isolates had 13 patterns, and these patterns were different from each other, except for 1 pattern . As a result, the PFGE patterns in 14 of 18 pairs were indistinguishable and those in 4 pairs were different . These data indicate a possible high rate of intrafamilial transmission of nontypeable H . influenzae, including BLNAR strains, between children and their parents.

Clin Diagn Lab Immunol, 2004 Jan, 11(1), 89 - 93
Measurement of serum bactericidal activity specific for Haemophilus influenzae type b by using a chromogenic and fluorescent metabolic indicator; Romero-Steiner S et al.; We evaluated alamarBlue as a metabolic indicator in a standardized assay for the measurement of serum bactericidal activity (SBA) to Haemophilus influenzae type b (Hib) using sera containing natural and vaccine-induced anticapsular (polyribosylribitol phosphate) antibodies . SBA assays with a colorimetric and a fluorometric end point in the presence of alamarBlue were developed and compared to a standard SBA assay, where colony counts are performed to determine the titer (12) . A colorimetric end point required a spectrophotometer, whereas a fluorometric end point required a fluorometer . Prevaccination sera (n = 27) and postvaccination sera (n = 13) were tested by all three methodologies, and the SBA titers obtained in the presence of alamarBlue were compared to those from the standard method . Both the colorimetric and the fluorometric SBA titers were significantly correlated (r = 0.87 and r = 0.95, respectively) with those of the standard assay (>/= 50% killing as the SBA titer end point), and titers were not significantly different when compared to those of the standard assay (P > 0.68) . However, the fluorometric end point had superior performance and ease of titer determination compared to the colorimetric end point (95 versus 87% of SBA titers were within 2 dilutions of the standard titer) . Hib SBA assays with alamarBlue are reproducible, faster (same-day assay), and easier to perform than the standardized assay, which requires manual or automated colony counts . These semiautomated methodologies result in increased sample throughput and collection of data in digital formats that can be exported to data analysis programs for determination of SBA titers.

Arch Dis Child Fetal Neonatal Ed, 2004 Jan, 89(1), F57 - 60
Acellular pertussis vaccine given by accelerated schedule: response of preterm infants; Slack MH et al.; OBJECTIVE: To describe the immune response of preterm infants to a diphtheria/tetanus/three component acellular pertussis (DTaP) vaccine, under an accelerated schedule, and the effects of steroids on this response . To compare responses with those of term infants . DESIGN: Prospective observational study . SETTING: Five Wessex neonatal units; Hertfordshire immunisation clinics . PATIENTS: Infants born at < 32 weeks; term controls . INTERVENTIONS: DTaP-Haemophilus influenzae type b vaccine given at 2, 3, and 4 months . Blood taken to assess antibody responses to vaccines . MAIN OUTCOME MEASURES: IgG geometric mean concentrations (GMC) to vaccines . RESULTS: A total of 130 preterm (mean gestational age 29.1 weeks) and 54 term infants were recruited . After the third immunisation, preterm infants had similar GMCs to controls to diphtheria, tetanus, filamentous haemagglutinin (FHA), and pertactin (PRN), but a significantly lower GMC to pertussis toxin (PT) . Responses to tetanus and PRN increased with age at the third immunisation, and those to tetanus, FHA, PRN, and PT increased with gestational age at birth . Response to tetanus correlated negatively with the number of doses of antenatal steroids received . There was no association between responses and postnatal steroids . CONCLUSION: When immunised with a combined acellular pertussis- H influenzae type b vaccine under an accelerated schedule, IgG GMC of preterm infants to PT was reduced . GMCs to tetanus, FHA, PRN, and PT increased with gestational age at birth, and GMCs to tetanus and PRN increased with age at the third immunisation . There is, however, no benefit in delaying immunisation . Anti-tetanus IgG decreased with increasing number of doses of antenatal steroids . There was no effect for postnatal steroids.

Expert Rev Vaccines, 2003 Oct, 2(5), 633 - 48
Conjugate vaccines; Lockhart S; Conjugate vaccines in which the capsular polysaccharide of Haemophilus influenzae type b, common serotypes of Streptococcus pneumoniae and group C Neisseria meningitidis are covalently bound to a protein antigen to convert a T-cell-independent immune response into a T-cell-dependent response have proved highly effective in the prevention of invasive disease in infants and young children . This review looks at what has been learnt from developing these vaccines that could be useful for the clinical development of future conjugate vaccines, in such areas as combination vaccines, dose-ranging, cross-protection of closely related antigens, prevention of noninvasive disease, correlates of protection and use in older age groups . In addition, a wide range of other organisms may be susceptible to conjugate vaccines and new indications and approaches are considered . This will be a highly active area for many years to come.

Expert Rev Vaccines, 2003 Oct, 2(5), 619 - 31
Heptavalent pneumococcal conjugate vaccine: current and future impact; Hsu KK et al.; Diseases caused by Streptococcus pneumoniae contribute significantly to worldwide morbidity and mortality . S . pneumoniae is now the number one cause of invasive bacterial disease in children in countries where Haemophilus influenzae type b (Hib) disease has been conquered by use of the Hib conjugate vaccine . Licensure of a heptavalent pneumococcal conjugate vaccine for prevention of invasive pneumococcal disease in children less than 5 years old in the USA represents the culmination of more than five decades of pneumococcal vaccine development . This review will: highlight safety, immunogenicity and efficacy studies that led to US Food and Drug Administration approval of this PCV7; summarize data about the incidence of childhood pneumococcal disease in the USA subsequent to licensure; review PCV7 treatment guidelines currently in place in the USA, Canada and Australia; and consider future directions in pneumococcal vaccine research.

Laryngoscope, 2004 Jan, 114(1), 129 - 31
Microbiology of recurrent acute rhinosinusitis; Brook I et al.; OBJECTIVE: We undertook to evaluate the microbiology of recurrent acute rhinosinusitis . METHODS: Repeated aspirations of maxillary sinus secretions by endoscopy were performed in eight patients over a period of 98 to 185 days . RESULTS: Bacteria were recovered for all 25 aspirates . A total of 31 isolates-14 Streptococcus pneumoniae, 11 Haemophilus influenzae, 5 Moraxella catarrhalis, and 1 Staphylococcus aureus-were recovered . The organism persisted in consecutive cultures in 13 instances and were eliminated in 8, and new organisms emerged in 6 instances . An increase in antimicrobial resistance was noted in 5 instances (3 in S . pneumoniae and 2 H . influenzae) . CONCLUSIONS: This study illustrates the microbial dynamics of recurrent acute rhinosinusitis, with the changes in microbial findings and increased bacterial resistance that occurs over time.

J Exp Med, 2004 Jan 5, 199(1), 35 - 46
Granulocyte CEACAM3 is a phagocytic receptor of the innate immune system that mediates recognition and elimination of human-specific pathogens; Schmitter T et al.; Carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) are used by several human pathogens to anchor themselves to or invade host cells . Interestingly, human granulocytes express a specific isoform, CEACAM3, that participates together with CEACAM1 and CEACAM6 in the recognition of CEACAM-binding microorganisms . Here we show that CEACAM3 can direct efficient, opsonin-independent phagocytosis of CEACAM-binding Neisseria, Moraxella, and Haemophilus species . CEACAM3- but not CEACAM6-mediated uptake is blocked by dominant-negative versions of the small GTPase Rac . Moreover, CEACAM3 engagement triggers membrane recruitment and increased GTP loading of Rac that are not observed upon bacterial binding to CEACAM6 . Internalization and Rac stimulation are also inhibited by compromising the integrity of an immunoreceptor tyrosine-based activation motif (ITAM)-like sequence in the cytoplasmic tail of CEACAM3 or by interference with Src family protein tyrosine kinases that phosphorylate CEACAM3 . In contrast to interfering with CEACAM6, blockage of CEACAM3-mediated events reduces the ability of primary human granulocytes to internalize and eliminate CEACAM-binding bacteria, indicating an important role of CEACAM3 in the control of human-specific pathogens by the innate immune system.

Clin Microbiol Infect, 2004 Jan, 10(1), 27 - 36
Clinical and bacteriological efficacy of the ketolide telithromycin against isolates of key respiratory pathogens: a pooled analysis of phase III studies; Low DE et al.; A pooled analysis of data from 13 phase III studies of telithromycin in the treatment of community-acquired pneumonia, acute exacerbations of chronic bronchitis, acute sinusitis or group A beta-haemolytic streptococcal pharyngitis and tonsillitis was undertaken . Causative key respiratory tract pathogens (Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Staphylococcus aureus and Streptococcus pyogenes) were isolated at entry to the studies from cultures of relevant respiratory samples and tested for their susceptibility to telithromycin, penicillin and macrolides (erythromycin A) . The combined clinical and bacteriological efficacy of telithromycin at the post-therapy, test-of-cure visit (days 17-24) was assessed in patients from whom a microbiologically evaluable pathogen was isolated at entry . More than 98% of key respiratory pathogens isolated, including penicillin G- and macrolide (erythromycin A)-resistant strains of S . pneumoniae, demonstrated full or intermediate susceptibility to telithromycin in vitro at the breakpoints of < or = 1.0 mg/L (susceptible) and 2.0 mg/L (intermediate) used for the purpose of evaluating the susceptibility of isolates recovered during the clinical trials . Treatment with telithromycin 800 mg once-daily for 5, 7 or 7-10 days resulted in high rates of clinical cure (88.5%) and a satisfactory bacteriological outcome (88.9%), similar to the figures seen with comparator antibacterial agents . Clinical cure and eradication rates were good for all key respiratory pathogens, including penicillin G- and macrolide (erythromycin A)-resistant S . pneumoniae . The results suggest that telithromycin will provide effective empirical therapy for community-acquired upper and lower respiratory tract infections.

Med Dosw Mikrobiol, 2003, 55(3), 231 - 43
{Polymorphism analysis of Haemophilus influenzae strains isolated from healthy children with respect to amplification reaction}; Nowaczek A et al.; The random-amplified polymorphic DNA (RAPD) assay was used to generate DNA fingerprints for 44 isolates H . influenzae obtained from healthy children . Problems with reproducibility and discriminatory power, frequently cited in the literature, were overcome by optimization procedure allowing to achieve reliable conditions for H . influenzae analysed . Particular parameters of RAPD fingerprinting were evaluated with respect to selection of best working primer, DNA polymerase and DNA concentration for amplification pattern . This study proved high sensitivity and efficiency of optimized RAPD profiling applicable for searching the epidemiology traces.

Pediatrics, 2004 Jan, 113(1 Pt 1), e40 - 6
Topical ciprofloxacin/dexamethasone otic suspension is superior to ofloxacin otic solution in the treatment of children with acute otitis media with otorrhea through tympanostomy tubes; Roland PS et al.; OBJECTIVE: To determine the efficacy and safety of topical ciprofloxacin/dexamethasone otic suspension compared with ofloxacin otic solution in the treatment of acute otitis media with otorrhea through tympanostomy tubes (AOMT) in pediatric patients . METHODS: This multicenter, prospective, randomized, observer-masked, parallel-group study was conducted at 39 sites in 599 children aged >or=6 months to 12 years with an AOMT episode of <or=3 weeks' duration . The mean age of patients was 2.5 years (standard deviation: 2.37 years) . Patients received either ciprofloxacin 0.3%/dexamethasone 0.1% otic suspension 4 drops twice daily for 7 days or ofloxacin 0.3% otic solution 5 drops twice daily for 10 days . Clinical signs and symptoms of AOMT were evaluated at clinic visits on days 1 (baseline), 3 (on therapy), 11 (end of therapy), and 18 (test of cure) . A patient diary was used to measure time to cessation of otorrhea . Principal pretherapy pathogens included Streptococcus pneumoniae (16.8%), Staphylococcus aureus (13.0%), Pseudomonas aeruginosa (12.7%), Haemophilus influenzae (12.4%), S epidermidis (10.2%), and Moraxella catarrhalis (4.1%) . RESULTS: Ciprofloxacin/dexamethasone is superior to ofloxacin for clinical cure (90% vs 78%) and microbiologic success (92% vs 81.8%) at the test-of-cure visit, produces fewer treatment failures (4.4% vs 14.1%), and results in a shorter median time to cessation of otorrhea (4 days vs 6 days) . Ciprofloxacin/dexamethasone treatment is also superior to improvement in clinical response by visit, absence of otorrhea by visit, and reduction of otorrhea volume by visit . Both topical otic preparations are safe and well tolerated in pediatric patients . No change in speech recognition threshold or decrease in hearing from baseline, based on audiometric testing, was noted with either regimen . CONCLUSION: Topical ciprofloxacin/dexamethasone treatment is superior to topical ofloxacin in the treatment of AOMT.

J Biol Chem, 2004 Mar 19, 279(12), 11035 - 41 Epub 2003 Dec 31.
Vibrio cholerae thiol peroxidase-glutaredoxin fusion is a 2-Cys TSA/AhpC subfamily acting as a lipid hydroperoxide reductase; Cha MK et al.; Recently, novel hybrid thiol peroxidase (TPx) proteins fused with a glutaredoxin (Grx) were found from some pathogenic bacteria, cyanobacteria, and anaerobic sulfur-oxidizing phototroph . The phylogenic tree analysis that was constructed from the aligned sequences showed two major branches . Haemophilus influenzae TPx.Grx was grouped in one branch as a 1-Cys subfamily of the thiol-specific antioxident protein/AhpC family . Most TPx.Grx proteins, including Vibrio cholerae TPx.Grx, were grouped in the 2-Cys subfamily . To explain the existence of two subgroups in novel hybrid TPx proteins, we have compared the kinetics given by V . cholerae TPx.Grx, H . influenzae TPx.Grx, their separated TPx domains, and a set of mutants devoid of the redox-active cysteines . The kinetic study described here demonstrates clearly that V . cholerae TPx.Grx is a 2-Cys TPx subfamily . For the first time, we also demonstrate the lipid peroxidase activity of V . cholerae TPx.Grx fusion and suggest the in vivo function of 2-Cys TPx.Grx fusion serving as a lipid peroxidase.

J Biol Chem, 2004 Mar 26, 279(13), 12163 - 70 Epub 2003 Dec 29.
Physiological characterization of Haemophilus influenzae Rd deficient in its glutathione-dependent peroxidase PGdx; Pauwels F et al.; The chimeric peroxidase PGdx of Haemophilus influenzae Rd belongs to a recently identified family of thiol peroxidases capable of reducing hydrogen peroxide as well as alkylhydroperoxides by means of glutathione redox cycling . In the present study, we constructed a H . influenzae Rd strain, deficient in its PGdx encoding gene (open reading frame HI0572) . The mutant was shown by disk inhibition and liquid culture growth assays to exhibit increased susceptibility to organic hydroperoxides . The hampered growth was restored by complementing the interrupted gene on the genome with a replicating plasmid bearing an intact copy of the gene, hereby rejecting the possible influences of polar effects . Elevated levels of hydrogen peroxide scavenging activity, due to the catalase HktE, were measured in the absence of a functional pgdx gene rendering the mutant more resilient against hydrogen peroxide . On the other hand, after initiation of the stationary phase, aerobic cultures of the pgdx mutant were practically devoid of living cells, whereas wild-type counterparts retained viability . This observed feature was alleviated by complementation with the functional gene or with the addition of catalase.

Am J Prev Med, 2004 Jan, 26(1), 29 - 33
Underimmunization in Chicago children who dropped out of WIC; Cortese MM et al.; BACKGROUND: The Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) serves a large proportion of Chicago infants, but some discontinue participation before age 1 year . To determine if children who remained active at WIC immunization-linked sites after their first birthday were more likely to be immunized by ages 19 and 25 months than those who dropped out, a retrospective cohort study was conducted . METHODS: Four Chicago WIC sites that used monthly voucher pick-up were chosen . Children born from July 1, 1997 to September 30, 1997 who attended these sites were eligible (N=1142) . The cohort was divided into two groups: (1) active group (46%), who had a WIC visit on or after their first birthday; and (2) inactive group (54%), who had their last WIC visit before their first birthday . Children were enrolled through home visits . RESULTS: The records for 200 children were analyzed . By age 19 months, 65 (84%) of 77 active children had received one dose of measles-mumps-rubella vaccine (MMR), compared to 82 (67%) of 123 inactive children (risk ratio {RR}=1.3; 95% confidence interval {CI}, 1.1- 1.5) . By age 25 months, 64 (83%) active children had received four doses of diphtheria-tetanus-pertussis vaccine (DTP), one MMR, and three doses of Haemophilus influenzae type b vaccine (Hib), compared with 64 (52%) inactive children (RR=1.6; 95% CI, 1.3-2.0) . CONCLUSIONS: In this cohort, children active in WIC after their first birthday were more likely to be immunized by ages 19 and 25 months, compared with those who were no longer active . Chicago children who drop out of WIC may represent those at highest risk for underimmunization and may require special strategies to improve coverage.

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