Haemophilus

Clin Infect Dis, 2004 Jan 15, 38(2), 234 - 42 Epub 2003 Dec 19.
Association of clinical signs and symptoms with bacterial findings in acute otitis media; Palmu AA et al.; In acute otitis media (AOM), a means of prediction of the bacterial pathogen based on symptoms and signs would be valuable in selecting appropriate antimicrobial treatment . Children in the control arm (n=831) in the Finnish Otitis Media Vaccine Trial were prospectively observed in a study clinic setting from the age of 2 to 24 months . In patients with AOM, myringotomy with aspiration was performed, and middle ear fluid samples were cultured for bacterial pathogens . Symptoms and signs of respiratory infections were thoroughly recorded . Streptococcus pneumoniae, Moraxella catarrhalis, and Haemophilus influenzae were the most common bacterial pathogens . Pneumococcal AOM was associated with more-severe AOM characterized by fever and earache . AOM due to H . influenzae was associated with eye symptoms and findings . Accurate prediction of a bacterial cause of infection based on symptoms and signs of AOM was not possible, but a specific cause was predicted in some situations, with a high probability of applicability to clinical practice.

Microbiol Immunol, 2003, 47(12), 937 - 43
Molecular cloning and characterization of the HmrM multidrug efflux pump from Haemophilus influenzae Rd; Xu XJ et al.; We cloned a gene responsible for norfloxacin resistance from the chromosomal DNA of Haemophilus influenzae Rd, and designated the gene as hmrM . HmrM showed sequence similarity with NorM of Vibrio parahaemolyticus and YdhE of Escherichia coli and others that belong to the MATE family multidrug efflux pumps . The recombinant plasmid carrying the hmrM gene conferred elevated resistance not only to norfloxacin but also to acriflavine, 4 ', 6-diamidino-2-phenylindole, doxorubicin, ethidium bromide, tetraphenylphosphonium chloride, Hoechst 33342, daunomycin, berberine, and sodium deoxycholate in Escherichia coli KAM32, a drug-hypersensitive strain . We observed an Na+-dependent efflux of ethidium and an ethidium-induced efflux of Na+ in E . coli KAM32 cells harboring the plasmid carrying the hmrM gene . These results indicate that HmrM is an Na+/drug antiporter-type multidrug efflux pump . A difference in substrate preference was observed between HmrM, NorM, and YdhE.

Antimicrob Agents Chemother, 2004 Jan, 48(1), 203 - 8
In vitro pharmacodynamic activities of ABT-492, a novel quinolone, compared to those of levofloxacin against Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis; Gunderson SM et al.; ABT-492 is a novel quinolone with potent activity against gram-positive, gram-negative, and atypical pathogens, making this compound an ideal candidate for the treatment of community-acquired pneumonia . We therefore compared the in vitro pharmacodynamic activity of ABT-492 to that of levofloxacin, an antibiotic commonly used for the treatment of pneumonia, through MIC determination and time-kill kinetic analysis . ABT-492 demonstrated potent activity against penicillin-sensitive, penicillin-resistant, and levofloxacin-resistant Streptococcus pneumoniae strains (MICs ranging from 0.0078 to 0.125 micro g/ml); beta-lactamase-positive and beta-lactamase-negative Haemophilus influenzae strains (MICs ranging from 0.000313 to 0.00125 micro g/ml); and beta-lactamase-positive and beta-lactamase-negative Moraxella catarrhalis strains (MICs ranging from 0.001 to 0.0025 micro g/ml), with MICs being much lower than those of levofloxacin . Both ABT-492 and levofloxacin demonstrated concentration-dependent bactericidal activities in time-kill kinetics studies at four and eight times the MIC with 10 of 12 bacterial isolates exposed to ABT-492 and with 12 of 12 bacterial isolates exposed to levofloxacin . Sigmoidal maximal-effect models support concentration-dependent bactericidal activity . The model predicts that 50% of maximal activity can be achieved with concentrations ranging from one to two times the MIC for both ABT-492 and levofloxacin and that near-maximal activity (90% effective concentration) can be achieved at concentrations ranging from two to five times the MIC for ABT-492 and one to six times the MIC for levofloxacin.

Lancet Neurol, 2004 Jan, 3(1), 54 - 62
Adjunctive dexamethasone treatment in acute bacterial meningitis; Chaudhuri A; The initiation of antibiotic treatment on suspicion of bacterial meningitis is important, but it is not enough to improve the prognosis for patients, especially those with pneumococcal meningitis . The mortality and morbidity of pneumococcal meningitis are still devastating, and results of a recent randomised trial have shown evidence in favour of dexamethasone treatment given before or with the first antibiotic dose . Adjuvant dexamethasone is unequivocally recommended in children and adults with haemophilus meningitis or pneumococcal meningitis . The benefit of adjunctive dexamethasone is likely to be greatest in patients who are otherwise healthy and present early with acute bacterial meningitis . Dexamethasone is not currently recommended for the treatment of gram-negative bacillary meningitis and neonatal meningitis . Dexamethasone, before or with the first dose of antibiotic, is likely to be one of the most significant practice changes that will benefit many adults and children with common types of acute bacterial meningitis and has been of proven value in the developed world.

Ann Allergy Asthma Immunol, 2003 Nov, 91(5), 501 - 5
Severe T- and B-cell immune deficiency associated with malignant thymoma; Yel L et al.; BACKGROUND: Immunodeficiency with thymoma syndrome is a rare disorder that generally occurs after the fourth decade of life . Typically, thymic tumors are benign, and gradually developing immunodeficiency consists of hypogammaglobulinemia with low B-cell counts and an inverted CD4+/CD8+ T-cell ratio due to excessive CD8+ T cells . OBJECTIVE: To report the case of a 32-year-old, white man with an invasive malignant thymoma and profound combined T- and B-cell immunodeficiency associated with a normal CD4+/CD8+ T-cell ratio, absence of circulating B cells, and infection with an unusual organism . METHODS: The patient presented with a superior vena cava syndrome caused by a malignant thymoma . During chemotherapy and radiotherapy, he experienced recurrent episodes of pulmonary infections due to Haemophilus influenza and Serratia marcescens and persistent oral thrush . He was diagnosed as having thymoma and underwent immunological evaluation . RESULTS: Sixteen months after the diagnosis of thymoma, the immunological evaluation revealed profound lymphopenia, eosinopenia, very low counts of both CD4+ T cells and CD8+ T cells, and a normal CD4+/CD8+ ratio with negative delayed-type hypersensitivity skin test results . Hypogammaglobulinemia and absent specific antibody responses were associated with a lack of peripheral blood CD19+ B cells . Despite treatment with intravenous immunoglobulin, the patient died of respiratory insufficiency and sepsis secondary to a chronic pulmonary infection . CONCLUSIONS: Malignant thymoma may be associated with severe combined immunodeficiency . A normal CD4+/CD8+ ratio and the absence of peripheral B cells suggest a bone marrow defect that affects both T and B cells in the pathogenesis of this syndrome . Comprehensive immunological evaluation should be performed when thymoma is diagnosed to initiate an early and effective treatment to prevent life-threatening complications.

Ann Allergy Asthma Immunol, 2003 Nov, 91(5), 496 - 500
Coexistent yellow nail syndrome and selective antibody deficiency; Bokszczanin A et al.; BACKGROUND: Yellow nail syndrome (YNS) is a rare, often underdiagnosed condition of unknown origin . The clinical features of the syndrome include yellow nails, chronic sinusitis, bronchiectasis, pleural effusion, and lymphoedema . Despite the frequent occurrence of upper and lower respiratory tract infections in patients with YNS, comprehensive analysis of their humoral immunity has not been previously reported . OBJECTIVE: To present the case of a patient with YNS whose recurrent upper and lower respiratory tract infections may have been caused by an underlying selective antibody deficiency that manifests as impaired IgG antibody response to polysaccharide antigens . METHODS: The patient underwent cultures of purulent sputum for Streptococcus pneumoniae and Haemophilus influenzae, bronchial washings for H . influenzae, and nail scrapings for fungi . Her serum levels of IgG, IgA, IgM, IgG subclasses, and serum titers of IgG antitetanus toxoid, anti-H . influenzae, and anti-S . pneumoniae antibodies were measured . RESULTS: Cultures of purulent sputum were positive on multiple occasions for S . pneumoniae and H . influenzae and bronchial washings were positive for H . influenzae . Nail scrapings were consistently negative for fungi . She had no reductions in serum levels of IgG, IgA, IgM, or IgG subclasses and had normal serum titers of IgG antitetanus toxoid antibodies . However, she demonstrated impaired IgG antibody responses following immunization with Pneumovax and an H . influenza B vaccine . CONCLUSIONS: This case report describes the first comprehensive analysis of humoral immune function in a patient with YNS . The finding of a selective antibody deficiency in our patient provides a potential explanation for the occurrence of respiratory infections in YNS . Accordingly, we recommend that functional antibody determinations and quantitative serum immunoglobulins be evaluated in patients diagnosed as having this unusual, enigmatic syndrome.

Jpn J Antibiot, 2003 Oct, 56(5), 365 - 95
{Susceptibilities of bacteria isolated from patients with lower respiratory infectious diseases to antibiotics (2001)}; Shimada K et al.; From October 2001 to September 2002, we collected the specimen from 370 patients with lower respiratory tract infections in 16 institutions in Japan, and investigated the susceptibilities of the isolated bacteria to various antibacterial agents and antibiotics and patients' characteristics . Of 458 strains that were isolated from specimen (mainly from sputum) and assumed to be bacteria causing in inflammation, 456 strains were investigated . The breakdown of the isolated bacteria were: Staphylococcus aureus 69, Streptococcus pneumoniae 72, Haemophilus influenzae 85, Pseudomonas aeruginosa (non-mucoid) 44, P . aeruginosa (mucoid) 13, Klebsiella pneumoniae 32, Moraxella subgenus Branhamella catarrhalis 32, and others . Of 69 S . aureus strains, those with 4 micrograms/mL or more of MIC of oxacillin (methicillin-resistant S . aureus: MRSA) occupied 43.5% . Vancomycin and arbekacin showed the most potent activities against MRSA as observed in 2000 . The frequency of S . pneumoniae exhibiting low sensitivity to penicillin (penicillin-intermediate S . pneumoniae: PISP + penicillin-resistant S . pneumoniae: PRSP) was 59.7% and both rates of PISP and PRSP were the highest after 1992 . Carbapenems had strong activities against S . pneumoniae . Especially, panipenem and imipenem inhibited the growth of all 72 strains at 0.125 and 0.5 microgram/mL, respectively . Generally, all drugs had strong activities against H . influenzae with MIC90s of 16 micrograms/mL or less . The drug that had the strongest activity against H . influenzae was levofloxacin, which inhibited the growth of 80 of the 85 strains at 0.063 microgram/mL . Against P . aeruginosa mucoid strain, meropenem had a strong activity with MIC90 of 0.5 microgram/mL while, against non-mucoid strain, tobramycin had a strong activity with MIC90 of 2 micrograms/mL . K . pneumoniae showed good susceptibilities to all drugs except ampicillin and minocycline, and the MIC90s were 4 micrograms/mL or less . Particularly, cefmenoxime, cefpirome, and imipenem had the strongest activity (MIC90: 0.125 microgram/mL), and cefozopran had a strong activity, inhibiting the growth of all strains at 0.25 microgram/mL . Also, all drugs generally had strong activities against M . (B.) catarrhalis . MIC90s of all drugs were 4 micrograms/mL or less . The drug that had the strongest activity was minocycline and levofloxacin inhibiting all 32 strains at 0.063 microgram/mL . Most of the patients with respiratory infection were aged 70 years or older, accounting for approximately a half of the total (40.5%) . As for the incidence by the diseases, bacterial pneumonia and chronic bronchitis were the highest, being noted in 39.2% and 37.3% of all the patients, respectively . The bacteria frequently isolated from the patients with bacterial pneumonia were S . aureus (19.3%) and S . pneumoniae (19.9%) . In contrast, H . influenzae (22.0%) were frequently isolated from the patients with chronic bronchitis . Before the drug administration, the bacteria frequently isolated from the patients were S . pneumoniae (20.8%) and H . influenzae (21.5%) . S . pneumoniae and H . influenzae decreased after the initiation of drug administration while S . aureus increased . The isolation frequency of P . aeruginosa was higher after than before the initiation of drug administration . The bacteria were frequently isolated from the patients who had already treated with cephems were S . aureus and P . aeruginosa . From the patients who had already treated with macrolides, S . pneumoniae was the most frequently isolated while S . aureus was the most frequently isolated from the patients pre-treated with quinolones.

Clin Infect Dis, 2003 Dec 15, 37(12), 1593 - 9 Epub 2003 Nov 18.
High incidence of Haemophilus influenzae type b infection in children in Pacific island countries; Russell FM et al.; The Haemophilus influenzae type b (Hib) disease burden among children <5 years old in 4 Pacific island countries (PICs) was estimated . The incidence of confirmed Hib meningitis was calculated using the numbers of culture-confirmed isolates . In addition, the World Health Organization (WHO) Hib Rapid Assessment Tool (RAT) was used to estimate the true Hib meningitis incidence and the number of Hib meningitis and pneumonia cases, as well as the number of deaths due to Hib meningitis and pneumonia . The Hib meningitis annual incidence in 3 PICs was 70-84 cases per 100,000 children <5 years old . For PICs, the RAT is likely to overestimate the Hib pneumonia burden, as it assumes a 5 : 1 ratio of Hib pneumonia to Hib meningitis . The true ratio is likely to be 1 : 1 . The high Hib disease burden and the relative cost-effectiveness of Hib vaccine make the introduction of Hib vaccine a good investment for PICs, costing US1000 dollars-US10,000 dollars for each death prevented--a number that ignores savings from reductions in the cost of treatment.

Braz J Med Biol Res, 2004 Jan, 37(1), 151 - 8 Epub 2003 Dec 18.
Reimmunization after bone marrow transplantation--current recommendations and perspectives; Machado CM; Autologous and allogeneic bone marrow transplantation (BMT) recipients lose immune memory of exposure to infectious agents and vaccines accumulated through a lifetime and therefore need to be revaccinated . Diphtheria toxoid, tetanus toxoid, pertussis vaccine (children <7 years old), Haemophilus influenzae type b conjugate, 23-valent pneumococcal polysaccharide, inactivated influenza vaccine, inactivated polio vaccine and live-attenuated measles-mumps-rubella vaccine are the currently recommended vaccines to be included in a vaccination program after BMT . For most of them, the best time to vaccinate, the number of vaccine doses and/or the duration of immunity after vaccination have not been established . Vaccination protocols vary greatly among BMT centers, suggesting that the lack of sufficient data has not permitted the formulation of reliable recommendations . The use of other vaccines and the perspectives for different vaccination protocols are analyzed in this review.

J Pediatr (Rio J), 1995 Mar-Apr, 71(2), 62 - 6
{Vaccines against meningococcal disease}; Costa WA; The meningococcal disease has been a source of preoccupation all over the world . Epidemics have been registered periodically in developed or developing countries . The most frequent meningococci are those concerned with serogroups A, B and C.Researches to develop effective vaccines against the disease have been taking place since the 40's . Presently, only vaccines against the meningococci from the serogroups A, C, Y and W-135 are available . These vaccines have important limitations both due to the age range that they protect and the period of time that the protection lasts . They are not routinely used in vaccination programs and are indicated only in risky situations . This demands a strict epidemiological surveillance of the disease . There are no vaccines against the serogroup B meningococci with recognized effectiveness, although, in the last decades, many have been tested . There are perspectives of important advances in this area, mainly with the development of conjugated vaccines, like the Haemophilus influenzae type B vaccine . Until the present, the chemoprophylaxis of the intimate communicants of a case is the best way to avoid secondary cases.

J Pediatr (Rio J), 1994 Jul-Aug, 70(4), 197 - 205
{Haemophilus influenzae type b infections and its prevention by vaccines}; Martins RM; Haemophilus influenzae type b (Hib) infections and its prevention by vaccines are reviewed . Its prevalence is underestimated, due to technical pitfalls in the laboratory and to the difficulties of diagnosing respiratory infections, chiefly pneumonia . Taking into account all infections (respiratory, meningitis and others), Hib is probably the most important pathogen, compared to S . pneumoniae and N . meningitidis . The incidence of Hib meningitis in Brazil is presented and risk factors are discussed, as well as the preventive alternatives . The different Hib vaccines are analyzed and schedules of immunization with two of them (PRP-T and HbOC) are presented . Possible eradication of Hib diseases is discussed.

Pediatr Infect Dis J, 2003 Dec, 22(12), 1063 - 8
Microbiology of otitis media in Costa Rican children, 1999 through 2001; Arguedas A et al.; BACKGROUND: Because of the increasing number of resistant middle ear pathogens and the impact of the new conjugate Streptococcus pneumoniae vaccine, an active surveillance of the microbiology and susceptibility pattern of middle ear pathogens is required . OBJECTIVE: To study the microbiology and susceptibility pattern of middle ear pathogens obtained from Costa Rican children with acute otitis media (AOM), recurrent otitis media (ROM) and therapeutic failure otitis media (FOM) . METHODS: Between 1999 and 2001 middle ear fluid (MEF) was collected from 276 Costa Rican patients . S . pneumoniae serotyping and pulsed field gel electrophoresis analysis was done on available strains . RESULTS: Among the total study population, 102 were AOM patients, 98 were ROM patients and 76 were FOM patients . Overall S . pneumoniae (88 strains) was the most common pathogen isolated followed by Haemophilus influenzae (41 strains) and Streptococcus pyogenes (10 strains) . H . influenzae was the most common agent in FOM patients (P = 0.015) . Beta-lactamase production was observed in 3 of 41 (7%) H . influenzae strains and 3 of 3 (100%) Moraxella catarrhalis strains . Penicillin-nonsusceptible S . pneumoniae strains were more common in FOM (64%) and ROM (63%) patients than in AOM (42%) patients (P = 0.05) . S . pneumoniae serotype 19F was the most prevalent serotype, mainly within one distinct clone . CONCLUSIONS: Overall S . pneumoniae serotype 19F was the most common isolate from the middle ear fluid of Costa Rican children . Beta-lactamase-negative H . influenzae was the most prevalent in the subpopulation of patients with FOM . S . pyogenes was the third most common isolate and M . catarrhalis was uncommon.

Int J Pediatr Otorhinolaryngol, 2004 Jan, 68(1), 101 - 10
Eustachian tube gland tissue changes are related to bacterial species in acute otitis media; Caye-Thomasen P et al.; BACKGROUND AND OBJECTIVE: Prior investigations have shown that the number of mucus producing goblet cells in the middle ear and Eustachian tube (ET) mucosa is highly increased during and up to at least six months after experimental acute otitis media (AOM) caused by Streptococcus pneumoniae (SP) . Further, the volume of the mucus producing paratubal gland components is increased up to 3 months after the acute infection . These changes may in conjunction with a deteriorated ET function predispose a subsequent development of secretory otitis media . The present investigation compares changes in goblet cell density and gland structures of the ET during and after AOM caused by various bacteria typically encountered in this disease, with emphasis on potential differences due to bacterial species . METHODS: Rat models of AOM caused by SP, non-typeable or type b Haemophilus influenzae (NTHI/HIB) or Moraxella catarrhalis (MC) were studied longitudinally up to 6 months after bacterial challenge . The ET was dissected and decalcified, paraffin embedded and serially sectioned, followed by PAS/alcian blue staining . The goblet cell density and the paratubal gland composition and volume were determined morphometrically in every 20th section, using a light microscope . RESULTS: Regardless of bacterial species, the ET goblet cell density was increased from day 8 and peaked day 16, followed by some degree of normalisation, although not reaching normal numbers within the 6 month period, except for MC . The highest increase was seen in AOM caused by the non-typeable Haemophilus strain, followed by HIB, SP and MC . Except with MC, pathological intra-epithelial glands formed and goblet cells were found in mucosal areas normally devoid of these . In all species but MC, the volume of the paratubal glands progressed to peak 16 days post-inoculation, followed by a gradual normalisation . The volume was still increased 3 months after the acute infection, but completely normalised after 6 months . The increase was primarily due to hypertrophy of the mucous gland components and highest in AOM caused by the Haemophilus species, followed by SP . CONCLUSION: The Eustachian tube goblet cell density is increased during and up to at least six months after AOM regardless of bacterial species, except when employing MC, by which the density was increased for a few weeks only . Except in AOM caused by MC, the volume of the ET glands increases during and up to at least 3 months after infection, primarily due to hypertrophy of the mucous gland components . The non-typeable Haemophilus strain induced the highest increase of both goblet cell density and mucous gland volume . The increased secretory capacity of the ET following AOM may by excessive mucus secretion contribute to the deteriorated ET function found after AOM and thus predispose, sustain or aggravate middle ear disease.

Int J Pediatr Otorhinolaryngol, 2004 Jan, 68(1), 51 - 6
Alloiococcus otitidis in acute otitis media; Leskinen K et al.; OBJECTIVE: The bacterium Alloiococcus otitidis has been found to be associated with otitis media with effusion (OME) . When the culture method is used, its detection rate is low, whereas applying the polymerase chain reaction (PCR) yields significantly higher frequencies . This study was carried out to investigate the incidence of A . otitidis in children with acute otitis media (AOM) . METHODS: Multiplex PCR was used to detect A . otitidis together with Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae in the middle ear effusions (MEEs) of 118 children with AOM . The clinical outcome of AOM and the bacterial findings of MEEs were compared . RESULTS: A . otitidis was detected in 25% (30 of 118) of the tested MEE samples . Children over 2 years of age had significantly more often A . otitidis-positive MEEs (37%; 22 of 59) than younger children (14%; 8 of 59) (chi-square test, P<0.01) . There were no significant differences in the duration, clinical failures (after antibiotic treatment), or number of recurrences of AOM between the A . otitidis-positive and A . otitidis-negative children . CONCLUSIONS: A . otitidis is found from the MEEs of AOM . The present data suggest that it has no clinical significance in AOM, and it does not increase the risk of developing OME after AOM.

J Pediatr (Rio J), 1999 Jul, 75(Suppl 1), S149 - 54
{Immunization schedule: dynamics and updating}; Weckx LY et al.; OBJECTIVES: The objective of this article is to make an analysis of the dynamics of the imunization schedule and an updating of the practical aspects of the vaccination . METHODS: The authors, based on the official recommendations, in the imunization schedule of the Infectology Department of the Brazilian Society of Pediatrics and on their experience, present practical aspects to facilitate the understanding of the dynamics of application of the calendar . RESULTS: The current calendar of the Brazilian Society of Pediatrics (SBP) is presented with a practical analysis of the vaccines BCG, hepatitis B, poliomyelitis, Haemophilus influenzae type b (Hib), DPT and triple viral, which are also part of the Calendar of the National Program of Immunizations . Besides this, they analyze two other suitable vaccines for SBP, against varicella and hepatitis A . Finally they comment on the risk of urbanization of the yellow fever and the increasing indication of vaccination against this disease in Brazil . CONCLUSIONS: The imunization schedule should be dynamic, adapted to the epidemiologic characteristics of each country or place . The presented calendar is what is now recommended by the Infectology Department of the Brazilian Society of Pediatrics (SBP).

J Pediatr (Rio J), 1999 Jul, 75(Suppl 1), S46 - 56
{Bacterial meningitis - diagnosis and treatment}; Faria SM et al.; OBJECTIVE: To present new concepts on diagnosis and treatment of bacterial meningitis regarding etiologic agents, pathophysiology and options about antimicrobial, antiinflammatory and supportive therapy . METHODS: Bibliographic review from MEDLINE data including articles published during the last ten years . One classic article published before this period and chapters of textbooks on infectious diseases were also included . RESULTS: Initial empirical antibiotic therapy is chosen according to probable etiologic agents for the age group . In the CSF microbiological analysis, the gram stains can reveal bacteria in 50% to 80% of the cases and the culture in nearly 85% . The tests for detection of bacterial antigens are useful for the diagnosis but they present low sensitivity . The most common agents during the neonatal period continue to be E . coli, Streptococcus B and L . monocytogenes . Beyond this period, the incidence of meningitis by Haemophilus influenzae b had a significant decrease after the introduction of conjugate vaccines . However, S . pneumoniae and N . meningitidis continue to be frequent agents . Currently, the thirdgeneration cephalosporins, ceftriaxone or cefotaxime, are the antibiotic therapy of choice . They are used with ampicillin up to two months of life and alone beyond this age . Dexamethasone has showed to be effective in reducing the inflammatory response and the sequelae, mainly the auditory sequelae . The fluid restriction doesn't offer advantages in the supportive care . CONCLUSIONS: Early diagnosis and prompt treatment are related to good outcome . The new insights on pathophysiology, the new antibiotics and the increasing bacterial resistance have determined changes in treatment.

J Pediatr (Rio J), 1999 Nov, 75(Suppl 2), S177 - 84
{Clinical management of upper airway obstruction: epiglottitis and laryngotracheobronchitis}; Amantea SL et al.; OBJECTIVE: To present current concepts on diagnosis and treatment of upper airway obstruction, mainly related to differential diagnosis between acute viral laryngotracheobronchitis and epiglottitis.METHODS: Bibliographic review covering the last ten years, using both Medline system and direct research . The most relevant articles published about the subject were selected.RESULTS: Viral laryngotracheobronchitis is an acute self-limited disease of the upper airway in a child, clinically characterized by barking cough, stridor, hoarse voice, and upper respiratory symptoms . The disease is diagnosed by clinical signs and symptoms . Rarely, if no immediate airway management is needed, radiography of the neck may help to exclude other entities that cause laryngeal obstruction . In contrast to viral laryngotracheobronchitis, epiglottitis is characterized by inflammation of the supraglottic tissues and is caused mainly by Haemophilus influenzae type b . A previously healthy child suddenly develops a sore throat and fever . Within hours after the onset of symptoms the patient looks toxic, swallowing is painful and breathing is difficult . Drooling and cervical hyperextension are frequently present . Lateral neck radiograph is rarely required to the diagnosis and may delay appropriate management of the airway . Moderate viral laryngotracheobronchitis with stridor at rest and retractions should be treated with steroids (systemic or nebulized) and nebulized epinephrine . Severe viral laryngotracheobronchitis should be treated aggressively while arregements are made for endotracheal intubation . The diagnosis of epiglottitis requires immediate endotracheal intubation in the appropriate unit (emergency department, intensive care unit or surgical unit) and antimicrobial therapy . Alternatively at some medical centers children with severe upper airway obstruction have been treated with a mixture of helium and oxygen (70 to 80% concentration of helium) instead of room air or pure oxygen to avoid intubation.CONCLUSIONS: There are different levels of care for patients with upper airway obstruction, depending on their clinical presentation . The clinical manifestations of viral laryngotracheobronchitis may be confused with the presentation of epiglottitis . Despite this observation we believe that differential diagnosis between viral laryngotracheobronchitis and epiglottitis rests on clinical grounds.

J Pediatr (Rio J), 1997 Jan-Feb, 73(1), 26 - 31
{Prevalence of Haemophilus influenzae resistant to ampicillin, cefaclor, cefotaxime, chloramphenicol and cotrimoxazol isolated from laboratories in the city of São Paulo, Brazil}; Rey LC et al.; OBJECTIVE: To determine Haemophilus influenzae resistant to ampicillin and other antibiotics isolated from different clinical specimens . METHODS: Isolates of H . influenzae were identified by culture with V and X factors and the aminolevulinic test . Nitrocefin was used to detect beta-lactamase (betaLac) production isolates were tested for antimicrobial susceptibility by disc diffusion and agar dilution methods . Serotype b was assessed by slide co-agglutination . RESULTS: From 245 H . influenzae identified, 155 were tested for serotype b, 28% (43/155) of which were positive . The global rate of beta-lactamase-positive isolates was 9% (22/245) . Resistance was similar among serotype b (11.6%) and non-type b H . influenzae (9.8%) (p>0.05) . No difference on betaLac production was found according to specimen's origin or the patients' age . Resistances to other antibiotics (by agar dilution and disc diffusion method, respectively) were: chloramphenicol 3.3 to 7.1%; cefaclor: 1.6 to 3.9% and cotrimoxazol: 9.1 to 10.5% . No resistance to cefotaxime has been detected; 63% (5/8) beta-Lac-positive isolates by agar dilution showed also resistance to chloramphenicol, compared to 3% (4/118) in the ss-Lac-negative group (p<0.001) . CONCLUSIONS: H . influenzae ampicillin-resistance has shown to be lower than other hospital-based-studies in Sao Paulo, and comparable to rates found in healthy carriers . The association between ampicillin and chloramphenicol resistance was significant: where this pattern is frequently found, the initial therapy for severe H . influenzae infections - like meningitis - should include a third generation cephalosporin.

J Pediatr (Rio J), 1998 Jan-Feb, 74(1), 45 - 8
{Etiological profile of bacterial meningitis in a small hospital}; Elias ML et al.; OBJECTIVE: To determine the etiologic profile of cases of meningitis treated at a small hospital in Ribeirao Preto, State of Sao Paulo, and to compare it to those reported for other communities.METHODS: a retrospective study was conducted on 103 patients admitted from January 1992 to July 1996 with clinical and laboratory diagnosis of bacterial meningitis . The clinical criteria for diagnosis were based on patient history and physical signs and symptoms, and the laboratory criteria were based on cerebrospinal fluid examination (aspect, cytology, biochemistry, Gram staining, culture and countercurrent immunoelectrophoresis) and blood culture.RESULTS: The etiologic agent was identified in 81.5% of cases: Haemophilus influenzae type b in 32%, Neisseria meningitides in 25.2%, Streptococcus pneumoniae in 8.7%, Staphylococcus aureus and epidermidis in 8.7%, and others agents in 6.9% . CONCLUSIONS: The study showed that the cases of bacterial meningitis treated at the Santa Lydia Hospital of Ribeirao Preto are caused by the same agents detected in other places, with Haemophilus influenzae type b being the predominant one . The study also suggests that when many people are involved in the collection of material for laboratory tests, recovery of the agent may become difficult.

J Pediatr (Rio J), 2002 Nov-Dec, 78 Suppl 2, S195 - 204
{Vaccination and the respiratory tract--what should we know?}; Farhat CK et al.; OBJECTIVE: This article aims at presenting a review of the main vaccines used in the pediatric population with direct impact on the prevention of infectious processes of the respiratory tract in children . SOURCES: Data from articles published in national and international scientific journals . The data were selected by means of direct search or search in the Lilacs and Medline databases . SUMMARY OF THE FINDINGS: This article was structured in topics, presenting issues related to immunization against frequent pathogens of the respiratory tract in human beings: Haemophilus influenza, influenza virus, respiratory syncytial virus and Streptococcus pneumoniae . CONCLUSIONS: The prevention of respiratory diseases through immunization is one of the main measures for the control of respiratory infection . These new vaccines, especially against Haemophilus influenza, have been available for some years with acknowledged efficacy . Now, the vaccine against Streptococcus pneumoniae (heptavalent vaccine) leads to a new perspective towards the management of respiratory diseases in children . In risk populations, the control of viral diseases through immunization, actively against the influenza virus and passively against the syncytial respiratory virus, is also an efficient measure to reduce respiratory disease.

J Pediatr (Rio J), 2002 Nov-Dec, 78 Suppl 2, S129 - 40
{Etiological diagnosis of pneumonia--a critical view}; Rodrigues JC et al.; OBJECTIVES: To search literature related to the etiological diagnosis of acute pneumonia in children . SOURCES: Systematic review of Medline and Lilacs databases . SUMMARY OF THE FINDINGS: The use of new diagnostic methods such as immunological techniques and polymerase chain reaction has proven invaluable for specific diagnosis and epidemiological investigation, showing adequate sensitivity, specificity and promptness of results, with the aim of guiding therapy properly . Review of epidemiological studies of community acquired pneumonia showed that Streptococcus pneumoniae is still one of the most significant etiologic agents in all age groups, in developing and industrialized countries . Resistance of this agent to penicillin and cephalosporins is increasing in all continents and is worrisome . Atypical agents such as Mycoplasma pneumoniae and Chlamydia pneumoniae are common in community acquired pneumonia, mainly in children older than 4 years, representing one third of the cases in industrial countries . However, their prevalence in developing countries remain to be determined . Respiratory syncytial virus is also a very common etiology of community acquired pneumonia and may cause severe infections, mainly in infants and younger children . The introduction of new conjugated vaccines for Streptococcus pneumoniae and Haemophilus influenzae type b resulted in significant reduction of morbidity and mortality of pneumonia in children . CONCLUSIONS: A significant impact on morbidity and mortality of acute pneumonia in children is likely to occur if microbiological and antimicrobial control is continuously and dynamically performed, thus allowing for the development of new vaccines, particularly against the respiratory syncytial virus.

Anal Chem, 2003 Sep 15, 75(18), 4918 - 24
Linking mass spectrometry and slab-polyacrylamide gel electrophoresis by passive elution of lipopolysaccharides from reverse-stained gels: analysis of gel-purified lipopolysaccharides from Haemophilus influenzae strain Rd; Gulin S et al.; Haemophilus influenzae is an important cause of human disease, and its lipopolysaccharide (LPS) is known to be a major virulence factor . H . influenzae produces short-chain LPS of which the heterogeneity is often visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using silver staining for detection . Individual bands have not previously been recovered by this method in quantities sufficient for mass spectrometry . In an attempt toward the development of sensitive mass spectrometrical strategies to be used in structural studies of H . influenzae LPS and LPS from other bacteria, we have applied here our previously described slab-PAGE-based micropurification method to obtain unmodified LPS fractions of high purity (>95%) from a crude LPS preparation of H . influenzae strain Rd . Two LPS-fractions were obtained which, after a procedure including mild acid hydrolysis, dephosphorylation, and permethylation of the resulting oligosaccharides, were subjected to tandem electrospray ionization mass spectrometry (ESI-MS/MS) . The quantities of micropurified LPS fractions-the recovery of LPS in terms of total mass was 30%-were found sufficient to allow the characterization of LPS glycoforms . The ESI-MS spectra of the individual bands showed reduced heterogeneity . Furthermore, the integrity of the micropurified LPS was confirmed . The spectra-displayed molecular ions showed improved intensity, increased respective signal-to-noise ratios demonstrating the sensitivity of analysis . Consequently, both the direct determination of the molecular masses of the gel-separated LPS glycoforms and sequence analyses using ESI-MS/MS were possible.

Nippon Rinsho, 2003 Dec, 61(12), 2170 - 4
{Administration of acute exacerbation of chronic obstructive pulmonary disease}; Ito T et al.; We have to consider the exacerbation of chronic obstructive pulmonary disease(COPD) may be caused not only by infection, but also by acute exacerbation of chronic heart failure, pulmonary embolism, pneumothorax, or other cardiopulmonary complications . Because it is characteristic that the exacerbation of COPD is often recurensive, the most important thing is the administration during stable status . Approximately 40% of pathogens of the acute infectious exacerbation of COPD are Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis, Pseudomonas aeruginosa, Staphylococcus aureus, and Echelisia coli . Also, approximately 15% is exacerbated by atypical pathogens such as Chlamydia pneumoniae and approximately 30% is by viral infection . We should contemplate the possibility of pathogens according to the statistics, when we choose antibiotics empirically.

J Hepatol, 2004 Jan, 40(1), 31 - 9
Microbial mimics are major targets of crossreactivity with human pyruvate dehydrogenase in primary biliary cirrhosis; Bogdanos DP et al.; BACKGROUND/AIMS: Previous studies on patients with primary biliary cirrhosis (PBC) have shown extensive cross-reactivity between the dominant B- and T-cell epitopes of human pyruvate dehydrogenase complex-E2 (PDC-E2), and microbial mimics . Such observations have suggested microbial infection as having a role in the induction of anti-mitochondrial antibodies, through a mechanism of molecular mimicry . However the biological significance of these cross-reactivities is questionable, because PDC-E2 is so highly conserved among various species . METHODS: Interrogating protein databases, ten non-PDC-E2 microbial sequences with high degree of similarity to PDC-E2(212-226) were found in Escherichia coli (6), Helicobacter pylori, Pseudomonas aeruginosa, Cytomegalovirus, and Haemophilus influenzae . We report on a study testing reactivity and competitive cross-reactivity against these respective peptides, and in some cases the parent protein, using sera from 55 patients with PBC, compared to reactivity of 190 pathological and 28 healthy controls . RESULTS: Cross-reactivity to E . coli mimics was commonly seen in PBC, and in a subset of pathological controls except where there was no evidence of urinary tract infection and correlated with anti-mitochondrial reactivity . CONCLUSIONS: E . coli/PDC-E2 cross-reactive immunity characterizes primary biliary cirrhosis; the large number of E . coli immunogenic mimics may account for the dominance of the major PDC-E2 autoepitope.

Kyobu Geka, 2003 Dec, 56(13), 1126 - 9
{Acute purulent pericarditis due to Haemophilus influenzae treated successfully with subxiphoid pericardial drainage; report of a case}; Takabayashi S et al.; A surgical case of acute purulent pericarditis in 1-year 2-month old boy who developed pericardial tamponade rapidly is reported . A subxiphoid pericardiectomy through a median incision was performed . Haemophilus influenzae was isolated from the effusion . He recovered successfully with an administration of antibiotics after the drainage procedures and did not develop constructive pericarditis . Our experience suggested that early subxiphoid pericardial drainage was effective in treatment of acute purulent pericarditis.

Am J Med Sci, 2003 Dec, 326(6), 360 - 8
Current issues in the management of bacterial respiratory tract disease: the challenge of antibacterial resistance; Dunbar LM; The worldwide burden of respiratory tract disease is enormous . Resistance to penicillins, macrolides, and cephalosporins is now detected among the leading bacterial pathogens that cause respiratory tract infections (RTIs)-Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis . The increasing role of atypical/intracellular pathogens (eg, Chlamydia pneumoniae, Mycoplasma pneumoniae, Legionella pneumophila) in RTIs, as well as their increase in antibiotic resistance prevalence, continues to be of great concern . More recently introduced treatment options for RTIs include the newer respiratory fluoroquinolones, along with the macrolides and azalides . Although these agents demonstrate good activity against common respiratory pathogens, reduced susceptibility to these agents has been reported . The ketolides are recently developed antibacterial agents with targeted-spectrum activity against common respiratory tract pathogens, including atypical/intracellular pathogens, and a low potential for inducing resistance . These promising new drugs have shown in vitro and in vivo efficacy in the treatment of community-acquired RTIs, such as community-acquired pneumonia, acute exacerbations of chronic bronchitis, and acute bacterial maxillary sinusitis.

Carbohydr Res, 2003 Nov 14, 338(23), 2731 - 44
Structural profiling of lipopolysaccharide glycoforms expressed by non-typeable Haemophilus influenzae: phenotypic similarities between NTHi strain 162 and the genome strain Rd; Schweda EK et al.; Non-typeable Haemophilus influenzae (NTHi) is a significant cause of otitis media in children . We have employed single and multiple step electrospray ionization mass spectrometry (ESIMS) and NMR spectroscopy to profile and elucidate lipopolysaccharide (LPS) structural types expressed by NTHi strain 162, a strain obtained from an epidemiological study in Finland . ESIMS on O-deacylated LPS (LPS-OH) and core oligosaccharide (OS) samples of LPS provided information on the composition and relative abundance of glycoforms differing in the number of hexoses linked to the conserved inner-core element, L-alpha-D-Hepp-(1-->2)-{PEtn-->6}-L-alpha-D-Hepp-(1-->3)-L-alpha-D-Hepp-(1-->5)-{PPEtn-->4}-alpha-Kdop-(2-->6)-Lipid A of H . influenzae LPS . The strain examined was found to elaborate Hex2 to Hex5 LPS glycoform populations having structures identical to those observed for H . influenzae strain Rd {Risberg, A.; Masoud, H.; Martin, A.; Richards, J.C.; Moxon, E.R.; Schweda, E.K.H . Eur . J . Biochem . 1999, 261, 171-180}, the strain for which the complete genome has been sequenced . In addition, sialyllactose-containing glycoforms previously identified in strain Rd as well as several NTHi strains, were identified as minor components . Multiple step tandem ESIMS (MS(n)) on dephosphorylated and permethylated OS provided information on the arrangement of glycoses within the major population of glycoforms and on the existence of additional isomeric glycoforms . Minor Hex1 and Hex6 glycoforms were detected and characterized where the Hex6 glycoform was comprised of a dihexosamine-containing pentasaccharide chain attached at the proximal heptose residue of the inner-core unit . LPS structural motifs present in the NTHi strain 162 are expressed by a genetically diverse set of disease causing isolates, providing the basis for a vaccine strategy against NTHi otitis media.

Vaccine, 2004 Jan 2, 22(3-4), 378 - 82
Impact of routine vaccination with a conjugate Haemophilus influenzae type b vaccine; Hviid A et al.; Based on a unique nationwide registration of vaccinated children, we studied the impact of routine Hib vaccination with special emphasis on vaccine uptake and adherence, vaccine effectiveness with respect to Hib meningitis, and indirect effects with respect to Hib meningitis among the unvaccinated children . Uptake and adherence was generally satisfactory . We estimated >97% effectiveness for all three doses of vaccine and observed herd-immunity in unvaccinated children comparable to a vaccine effectiveness of 94% 3.5 years into the programme . In conclusion, nationwide routine Hib vaccination is highly effective in protecting against Hib meningitis, and rapid achievement of herd immunity is possible with catch-up vaccination of older children.

Eur J Clin Microbiol Infect Dis, 2004 Jan, 23(1), 46 - 8 Epub 2003 Dec 11.
Haemophilus parainfluenzae: an underdiagnosed pathogen of biliary tract infections?
Frankard J, Rodriguez-Villalobos H, Struelens MJ, Jacobs F.
Presented here is a case of monobacterial peritonitis complicating cholecystitis and caused by an uncommon agent of gastrointestinal infections, Haemophilus parainfluenzae . The pathogenic role of this organism in digestive infections, particularly in those of the biliary tract, has been reported increasingly though sporadically . Indeed, it has been shown to be a coloniser of the gastrointestinal tract, and a recent hypothesis of an ascending route of infection to the biliary tract has been postulated to partly explain its pathogenicity . More frequent identification of Haemophilus parainfluenzae as a causal agent of biliary tract infection would probably be obtained through the use of specific culture media, since its potential implication has been demonstrated.

J Pediatr Hematol Oncol, 2003 Dec, 25 Suppl 1, S28 - 33
Childhood chronic immune thrombocytopenic purpura: unresolved issues; Blanchette VS et al.; Chronic immune thrombocytopenic purpura (ITP), defined as a platelet count of below 150 x 109/L persisting for more than 6 months from onset of illness, occurs in approximately 20% to 25% of children with acute-onset ITP . A small subset of these patients (approximately 5%) will manifest symptomatic, severe thrombocytopenia (platelet counts <20 x 109/L) at 1 year or longer following diagnosis, and may require splenectomy . Complete/partial response rates following splenectomy in children with primary chronic ITP are of the order of 70% to 75%; response rates are lower in children with secondary ITP and those with complex autoimmune cytopenias (e.g., Evans syndrome) . Laparoscopic splenectomy is increasingly preferred over open splenectomy . Patients should be immunized with the pneumococcal, Haemophilus type b and meningococcal vaccines before splenectomy; the duration of postsplenectomy antibiotic prophylaxis using penicillin or an equivalent antibiotic is controversial but should be at least until 5 years of age and for a minimum of 1 year postsplenectomy . Some experts advocate life-long antibiotic prophylaxis . Treatment of postsplenectomy failures is a challenge; partial/complete remission rates are low, and multimodality therapy may be more efficacious than monotherapy . The presence of an accessory spleen should be sought and removal considered if present . The role of newer treatment modalities such as anti-CD 20 remains to be established.

J Infect, 2004 Jan, 48(1), 56 - 65
Antimicrobial susceptibility among community-acquired respiratory tract pathogens in the USA: data from PROTEKT US 2000-01; Doern GV et al.; OBJECTIVES: The PROTEKT US surveillance program (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin in the United States) commenced in 2000 to document the emergence and spread of antimicrobial resistance among respiratory tract pathogens in the United States . METHODS: During 2000-2001, 206 centers from 154 cities/metropolitan areas collected 16,727 clinical isolates (Streptococcus pneumoniae, n=10103, Streptococcus pyogenes, n=3918, Haemophilus influenzae, n=2706) . RESULTS: Among S . pneumoniae isolates, 38.9% showed decreased susceptibility to penicillin (12.5% intermediate, 26.4% resistant) with marked geographical variability . The erythromycin resistance rate was 31.0% and highly correlated with penicillin resistance . The rate of fluoroquinolone resistance was 0.8% . Telithromycin was nearly uniformly active against S . pneumoniae (MIC(90) 0.5 mg/l) . All isolates of S . pyogenes were penicillin-susceptible, 5.5% were resistant to erythromycin . Telithromycin minimum inhibitory concentrations (MICs) were lower than clindamycin and macrolide MICs against S . pyogenes (MIC(90) 0.03 mg/l versus 0.25 mg/l and 0.12 mg/l, respectively) . 28.3% of H . influenzae isolates produced beta-lactamase . Telithromycin activity versus H . influenzae was not affected by beta-lactamase production . CONCLUSIONS: The PROTEKT US study confirms the widespread prevalence of antimicrobial resistance among common bacterial respiratory pathogens in the US, and re-affirms the importance of continued surveillance to guide optimum empiric therapy for patients with Community-acquired respiratory tract infections (CARTIs) . The new ketolide, telithromycin, maintained potent activity against study isolates in vitro and offers promise for the effective treatment of CARTIs.

J Infect, 2004 Jan, 48(1), 39 - 55
Antibacterial resistance among children with community-acquired respiratory tract infections (PROTEKT 1999-2000); Felmingham D et al.; OBJECTIVE: To determine the susceptibility of bacterial respiratory tract pathogens, isolated from children (0-12 years) as part of the global PROTEKT surveillance study (1999-2000), to a range of antibacterials, including the ketolide telithromycin . METHODS: Minimum inhibitory concentrations of the antibacterials studied were determined at a central laboratory using the NCCLS microdilution broth method . Macrolide resistance mechanisms were detected by PCR . RESULTS: Of 779 Streptococcus pneumoniae isolates worldwide, 43% were non-susceptible to penicillin (18% intermediate; 25% resistant) and 37% were resistant to erythromycin, with considerable intercountry variation . Eighteen per cent of 653 Haemophilus influenzae and >90% of 316 Moraxella catarrhalis isolates produced beta-lactamase . Of 640 Streptococcus pyogenes isolates, 10% were resistant to erythromycin, with considerable intercountry variation . All S . pneumoniae and 99.8% of H . influenzae isolates were susceptible to telithromycin using breakpoints proposed to the NCCLS (<or=1 and <or=4 mg/L, respectively) . All M . catarrhalis and 97% of S . pyogenes and isolates were susceptible to <or=1 mg/L telithromycin . CONCLUSIONS: Antibacterial resistance complicates the empirical treatment of respiratory tract infections in children and requires continued monitoring . Telithromycin may be a useful therapeutic alternative as it is highly active against strains exhibiting various resistance phenotypes.

Eur J Haematol, 2003 Nov, 71(5), 319 - 26
Prevention of life-threatening infections due to encapsulated bacteria in children with hyposplenia or asplenia: a brief review of current recommendations for practical purposes; Castagnola E et al.; The aim of the present work was to summarise in a single paper all the options for prevention of life-threatening infections due to encapsulated bacteria in patients with hyposplenism or asplenia . Prevention of these infections should be obtained in all patients with 1) patient and family education, 2) prophylaxis by means of vaccination against Haemophilus influenzae and Streptococcus pneumoniae, 3) antibiotic prophylaxis, based primarily on penicillin, 4) delay of elective splenectomy or use methods of tissue salvage in splenic trauma . Vaccination is not effective against all serotypes of S . pneumoniae and Neisseria meningitidis causing life-threatening infections in hypo/asplenic patients . Moreover, antibacterial prophylaxis could select antibacterial-resistant pathogens and is highly conditioned by patient's compliance . Therefore, empirical antibacterial therapy of fever and/or suspected infection should be recommended to all splenectomised patients independently from time elapsing from splenectomy, vaccinal status and assumption of antibacterial prophylaxis.

Cad Saude Publica, 2003 Sep-Oct, 19(5), 1267 - 75 Epub 2003 Dec 02.
{Haemophilus influenzae type b: epidemiological situation in the State of Minas Gerais, Brazil, 1993-1997}; Miranzi Sde S et al.; Among Haemophilus influenzae type b (Hib) invasive diseases, pneumonia and meningitis are the most relevant in public health due to their frequency and severity . From 1993 to 1997, there were 720 cases of Hib meningitis in Minas Gerais State, Brazil, representing the most frequent cause of bacterial meningitis in infants (< 1 year) and the second most frequent among all causes of meningitis . The total estimated cases of invasive Hib diseases thus appear to justify the recent inclusion of the vaccine in the basic immunization protocol . The vaccine's high cost reinforces the need for more precise monitoring of the etiological diagnosis of meningitis cases, representing one of the weaknesses in the prevailing epidemiological surveillance system.

Respiration, 2003 Sep-Oct, 70(5), 507 - 14
Organizing pneumonia presenting as a solitary nodular shadow on a chest radiograph; Watanabe K et al.; BACKGROUND: The role of infection as a cause of focal organizing pneumonia (OP) is not fully understood . OBJECTIVES: This study aimed to determine the clinical, radiological and pathological characteristics of patients with OP presenting a solitary nodular shadow on a chest radiograph . METHODS: Fourteen patients who presented with a solitary nodular shadow on a chest radiograph, pathologically diagnosed as OP after surgical resection, were allocated into two groups according to the histological findings . The first had OP with aggregates of neutrophils in airways and/or pulmonary parenchyma with or without necrosis or destruction of lung architecture (group 1: OP with neutrophilic infiltration; n = 10) . The second consisted of patients with OP presenting neither neutrophilic infiltration, necrosis nor destruction of lung architecture (group 2: OP without neutrophilic infiltration; n = 4) . RESULTS: Cough, sputum, and chest pain were the common symptoms in both groups of patients . Computed tomography (CT) revealed that all nodules in both groups were located in the peripheral lung parenchyma and had irregular margins, and their shapes varied from round to wedge-shaped . Haemophilus influenzae was isolated from sputum or transbronchial aspirates from 3 patients in group 1 . CONCLUSIONS: The specimens resected from patients with focal OP mostly show small aggregates of neutrophils . However, the dominant histological feature is OP and neutrophilic infiltration, suggesting infection was a minor histological component in all cases . Clinical symptoms and chest CT findings did not clearly distinguish these two groups of patients . It is thus reasonable to place these OPs in the same category and to treat them in the same way . Surgical excision appears to be the only method for a precise diagnosis .

Ann Epidemiol, 2004 Jan, 14(1), 31 - 5
Invasive disease caused by Haemophilus influenzae: the sensitivity of statutory reporting; Dominguez A et al.; PURPOSE: The objective of the study was to investigate the sensitivity of the statutory reporting of Haemophilus influenzae invasive disease (HIID) during the period between 1996 and 1997 in Catalonia, Spain . METHODS: The incidence of HIID reported passively by clinicians was compared with the cases detected by a system of microbiological surveillance . In all cases isolated, the age of the patient, the clinical form and the serotype were investigated . RESULTS: Sixty-six cases were passively reported and 111 were detected by microbiological surveillance . Overall sensitivity of reporting was 59.5% (95% CI, 50.7-69.1), with differences being observed according to the variables studied . The highest values were obtained in children under 5 years (87.8%; 95% CI, 73.7-95.9), in cases with meningitis and/or sepsis (64.3%; 95% CI, 50.4-76.6) and in serotype b (73.0%; 95% CI, 60.3-83.4) . In the logistical regression analysis, only age under 5 years was associated with greater reporting (OR= 9.8; 95% CI, 2.5-37.8) . CONCLUSIONS: Reported morbidity in children under 5 years is a good estimate of the true incidence of HIID in Catalonia (underreporting was 12.2%), but not for those above this age in whom the underreporting was 57.2%.

Presse Med, 2003 Nov 22, 32(37 Pt 1), 1752 - 9
{Bacteria and resistance to antibiotics in acute otitis media in paediatrics, depending on the geographical origin}; Bidet P et al.; EPIDEMIOLOGY OF THE BACTERIA RESPONSIBLE: Acute otitis media (AOM) is the most common bacterial infection in childhood below the age of 5 years . Bacteria may be isolated from middle ear fluid in about two-thirds of patients . The prevalence of bacteria varies from one country to the next . The most common pathogens recovered are Streptococcus pneumoniae, Haemophilus influenzae (20-50%) and less frequently Moraxella catarrhalis (10%) . However, several recent reports suggest an increasing rate of isolation of M . catarrhalis approaching 20% . Concomitant isolation of two or more organisms occurs in up to 10% of cases . The role of Group A Streptococci and Staphylococcus aureus in AOM has decreased since the use of antibiotics . EPIDEMIOLOGY OF ANTIMICROBIAL RESISTANCE: The recent spread of penicillin resistant S . pneumoniae and amoxycillin resistant H . influenzae varies considerably from one country to an other and appears related to the use of antibiotics and socio-economic conditions . A follow up of bacterial epidemiology and antibiotic resistance is necessary in each region of the world to define accurate strategies of acute otitis antibiotherapy.

J Clin Microbiol, 2003 Dec, 41(12), 5546 - 50
Evaluation of methodology for serotyping invasive and nasopharyngeal isolates of Haemophilus influenzae in the ongoing surveillance in Brazil; Bokermann S et al.; To assess the magnitude of discrepant results obtained by routine Haemophilus influenzae serotyping, 258 isolates, collected by the epidemiological surveillance system in Brazil from individuals with invasive diseases or carriage, were evaluated by two slide agglutination (SlAg) methods: SlAg method 1, by which strains were initially screened with a serotype b-specific antiserum, and SlAg method 2, by which strains were tested against all serotype-specific antisera in parallel . Investigators comparing results of the two SlAg methods with those obtained by capsule type-specific PCR were blinded to the method used . The serotype prevalence rates found by the three methods were significantly different, involving discrepancies mainly between serotype b and noncapsulated (NC) isolates . For invasive isolates (n = 131), the overall agreement rate between SlAg method 1 or 2 and PCR was 68.0 or 88.3%, respectively, whereas for colonizing isolates (n = 127) the corresponding rate was 46.5 or 94.2%, respectively . SlAg method 2 improved the ascertainment of serotypes over that obtained with SlAg method 1, demonstrating good correlation with PCR . Use of the polyvalent antiserum as a screening reagent for SlAg for invasive and colonizing isolates showed poor discriminatory power, with a sensitivity of 65.8% and a specificity of 91.7% . We stress the importance of using a well-standardized SlAg methodology and suggest that reference laboratories should utilize PCR routinely to confirm SlAg results and to check all nonspecific SlAg reactions and apparent NC isolates by SlAg in order to provide reliable data on the prevalence of H . influenzae serotypes in the H . influenzae type b vaccine era.

Int J Antimicrob Agents, 2003 Dec, 22(6), 557 - 61
In vitro activities of nine peptide deformylase inhibitors and five comparator agents against respiratory and skin pathogens; Bowker KE et al.; The activity of nine peptide deformylase (PDF) inhibitors undergoing clinical evaluation were compared with co-amoxiclav, levofloxacin, moxifloxacin, erythromycin and telithromycin against a range of respiratory and skin pathogens (n=166) . The PDF inhibitor showed good activity against Streptococcus pneumoniae, Moxarella catarrhalis, Group A streptococci and Staphylococcus aureus irrespective of beta-lactam or fluoroquinolone susceptibility . Against Haemophilus influenzae, MIC(90) values were generally higher . BB-88488 was the most active compound . Overall these data suggest that PDF inhibitors are an interesting new class of antimicrobial worthy of further investigation in the treatment of respiratory tract and skin infections.

P N G Med J, 2002 Mar-Jun, 45(1-2), 44 - 50
Pneumonia vaccine trials at Tari; Riley ID; Pneumonia is the commonest cause of death of children in Papua New Guinea (PNG) . At Tari pneumonia is most commonly caused by Streptococcus pneumoniae and Haemophilus influenzae, which set up rapid severe infections in the lungs that require urgent treatment . In rural PNG, however, treatment is often delayed . Penicillin-resistant forms of these bacteria are on the increase . It is therefore important to have another means of protection against this serious disease . This paper describes three field trials of a vaccine against the commonest serotypes of S . pneumoniae found in PNG . The trials show that a pneumococcal vaccine can prevent deaths from uncomplicated acute lower respiratory tract infection in small children and adults . It is likely that the vaccine does this by limiting the replication of bacteria in the lungs and thus limiting their spread to other parts of the body.

J Pediatr, 2003 Dec, 143(6 Suppl), S163 - 87
Global reduction of Hib disease: what are the next steps? Proceedings of the meeting Scottsdale, Arizona, September 22-25, 2002; Watt JP et al.; On September 22 to 25, 2002, a group of infectious disease specialists, public health officials, and vaccine experts from 33 countries gathered in Scottsdale, Arizona, to discuss the epidemiology and control of disease caused by Haemophilus influenzae type b (Hib) in the era of Hib conjugate vaccines . This supplement is a synthesis of the major themes and key lessons identified at the meeting . The objectives of the conference were to review the 10-year experience with Hib conjugate vaccines, discuss strategies to reduce Hib disease rates to lowest possible levels in industrialized countries, review impediments to the introduction of Hib vaccine in developing countries, and discuss strategies for disseminating lessons learned from countries using to those not using Hib conjugate vaccines . Over 10 years of international experience with Hib conjugate vaccines has demonstrated that they are safe and effective . Routine use of Hib conjugate vaccine has consistently led to decreases in the incidence of invasive Hib disease of 90% or more across a wide range of epidemiologic situations in industrialized countries . In some countries, the vaccine has caused a near-disappearance of invasive Hib disease through a combination of direct protection and herd immunity . Developing countries that have implemented routine vaccination (eg, The Gambia, Chile) have also had substantial disease reduction . In countries where Hib conjugate vaccine is being used, reducing Hib disease incidence to the lowest possible level will depend on maintaining high vaccine coverage levels, conducting surveillance for Hib disease, and investigating Hib disease cases . The optimal Hib vaccination strategy will depend on many factors, including local epidemiology and programmatic considerations . In countries that are not using Hib conjugate vaccine, information on the local burden of Hib disease will be essential for leaders considering vaccine introduction . Where disease burden is high, a multifaceted approach is urgently needed to evaluate and overcome barriers to vaccine introduction . In areas where Hib disease burden is not well characterized, additional work will be needed to understand the epidemiology of Hib disease and to communicate the value of Hib conjugate vaccine.

Genome Res, 2003 Dec, 13(12), 2665 - 73
Hierarchy of sequence-dependent features associated with prokaryotic translation; Lithwick G et al.; Protein expression in the cell is affected by various sequence-dependent features . Several such sequence-dependent features have been individually studied,yet they have not been compared quantitatively in terms of their relative influence on protein expression,and a hierarchy of these elements has not been determined . Here we present a quantitative analysis examining sequence-dependent features involved in prokaryotic translation,namely,the base-pairing potential between the mRNA Shine-Dalgarno sequence and the ribosomal RNA,codon bias,and the identity of the stop codon . We analyzed these features both at intra- and intergenomic levels using the Escherichia coli and Haemophilus influenzae genomes . Within each genome,we examined the relationship between each feature and protein expression levels determined by 2D-gel analyses . At the intergenomic level,comparative genomic principles were applied to study the relative preservation of the different sequence-dependent properties between orthologs . From these analyses,we determined that biased codon usage is the property that is most highly associated with protein expression and that is most conserved . The identity of the stop codon and the base-pairing potential of the mRNA Shine-Dalgarno sequence and the rRNA seem to have less of an effect on protein expression.

Eur J Pediatr, 2004 Feb, 163(2), 105 - 7 Epub 2003 Dec 04.
The aetiology of paediatric inflammatory vulvovaginitis; Cuadros J et al.; Vulvovaginitis is the most common gynaecological problem in prepubertal girls and clear-cut data on the microbial aetiology of moderate to severe infections are lacking . Many microorganisms have been reported in several studies, but frequently the paediatrician does not know the pathogenic significance of an isolate reported in vaginal specimens of girls with vulvovaginitis . A multicentre study was performed, selecting 74 girls aged 2 to 12 years old with a clinical picture of vulvovaginitis and inflammatory cells on Gram stain . All the specimens were cultured following standard microbiological techniques and the paediatricians completed a questionnaire to highlight risk factors after interviewing the parents or tutors . The data were compared with those obtained in a control group of 11 girls without vulvovaginitis attending a clinic . Streptococcus pyogenesand Haemophilus spp.were isolated in 47 and 12 cases, respectively . Upper respiratory infection in the previous month ( P<0.001) and vulvovaginitis in the previous year ( P<0.05) were identified as significant risk factors . Foreign bodies, sexual abuse, poor hygiene and bad socioeconomic situation were not identified as risk factors for the infection . CONCLUSION: Paediatric inflammatory vulvovaginitis is mainly caused by pathogens of the upper respiratory tract and the most common risk factor for this infection is to have suffered an upper respiratory tract infection in the previous month.

Pediatrics . 2003 Dec;112(6 Pt 1):e453.
Injection-site reactions to booster doses of acellular pertussis vaccine: rate, severity, and anticipated impact; Skowronski DM et al.; BACKGROUND: Acellular pertussis (aP)-containing vaccines cause fewer adverse events than whole-cell versions for primary doses . Booster doses, however, may be followed by extensive injection-site reactions . This study compares the frequency, severity, and impact of local reactions among children receiving 5 consecutive doses of an aP combination vaccine (including inactivated polio virus, conjugated Haemophilus influenzae type b antigen, and diphtheria and tetanus toxoids) to children receiving a mixed series of whole-cell and aP combination vaccines . METHODS: Participants were parents or guardians of children 4 to 6 years old immunized at public health clinics across British Columbia, Canada . This included 398 children receiving the fifth consecutive dose of an aP combination vaccine and 402 receiving the fifth dose in a mixed series consisting of at least 1 prior dose of whole-cell pertussis combination vaccine with the remainder as aP combination vaccine . A cross-sectional telephone survey evaluated the extent of local reactions 48 to 96 hours after immunization by asking participants to compare the size of redness and swelling with familiar household items such as Oreo cookies or coins . Associated discomfort and impact on recreational activities, health care utilization, parental time off work, and attitudes toward immunization were also assessed . RESULTS: Children who received the fifth consecutive dose of an aP combination vaccine more often experienced redness (24%) or swelling (16%) the size of an Oreo cookie or larger (>or=46 mm) than children given a mixed series (10% and 9%, respectively) but less often experienced tenderness or limitation of movement at the injection site . Related health care utilization was low . There was no discernible effect on participation in recreational activities or parental attitudes toward vaccine; 90% would recommend the same vaccine to others with children of the same age . CONCLUSIONS: We conclude that injection-site reactions are more extensive after the fifth consecutive dose of an aP combination vaccine compared with the fifth dose in a mixed series of whole-cell and aP combination vaccines . These reactions are unlikely to affect parental acceptance of immunization recommendations or health care utilization.

Hunan Yi Ke Da Xue Xue Bao, 2003 Jun, 28(3), 272 - 4
{Characteristics of bacteriology and drug sensitivity in patients with COPD combined with pneumonia}; Yi ZM et al.; OBJECTIVE: To investigate the characteristics of bacteriology and the drug resistance in the patients with chronic obstructive disease (COPD) combined with pneumonia at our respiratory intensive care unit(RICU) in 2001 . METHODS: The sputum of 543 patients with COPD combined with pneumonia was collected with a regular method, the bacteria were cultured, and then the drug sensitivity was analyzed with K-B method . RESULTS: There were 181 positive samples in the 543 patients with the positive rate 33.9% . The main bacteria in the G- bacillus were Pseudomonas (21.55%), Haemophilus influenza (14.36%), Enteric bacilli (13.26%), and Klebsiella (11.60%) which were sensitive to impipenem, amikacin, and ceftazidime . The main bacteria in G+ bacteria were Staphylococcus aureus (7.18%) and Staphylococcus epidermindis (4.42%) which were sensitive to vancomycin, imipenem, ceftazidime, and amikacin . CONCLUSION: The bacteria in the patients with COPD combined with pneumonia at RICU of the hospital in 2001 are mainly made up of G- bacillus and secondarily of G+ bacteria which are sensitive to imipenem, amikacin, and ceftazidime . The G+ bacteria are highly sensitive to vancomycin.

J Pediatr (Rio J), 2001 Sep-Oct, 77(5), 387 - 92
{Haemophilus influenzae type b meningitis in the state of Paraná, Brazil}; Takemura NS et al.; OBJECTIVE: During the second half of 1996, the municipalities of Londrina and Curitiba (State of Parana, Brazil) included Haemophilus influenzae type b (Hib) vaccine into their routine vaccination regimen, approximately 30 months before its introduction into the National Immunization Program . The present study aimed at verifying the incidence of meningitis caused by Hib among children in Londrina, Curitiba, and in the remaining municipalities of the State, before and after the introduction of this vaccine into the immunization program . METHODS: An observational and retrospective study was carried out . The study included all cases of Haemophilus influenzae type b meningitis recorded by the epidemiological surveillance system in Londrina and by the State of Parana Health Secretariat between 1992 and 1999 among children aged less than 5 years . The incidence rates of Hib meningitis were calculated per 100,000 children aged less than five years . RESULTS: After the introduction of Hib vaccine, an important reduction in the incidence rate of Haemophilus influenzae type b meningitis was observed in Londrina (from 23.91 in 1996 to 2.79 in 1999) . A Similar decrease was observed in Curitiba . In the remaining localities of the state, which had not introduced the vaccine till mid-1999, the incidence rate remained almost unchanged . CONCLUSION: Regular vaccination against Hib was effective in reducing the incidence rate of meningitis amongst children younger than five years in Londrina and Curitiba . In order to maintain this low incidence rate, adequate vaccination coverage and strict epidemiological surveillance should be guaranteed.

J Pediatr (Rio J), 2002 Jan-Feb, 78(1), 24 - 30
{Etiology and evolution of bacterial meningitis in a pediatric center}; Romanelli RM et al.; OBJECTIVE: To establish the prevalence of the etiological agents of bacterial meningitis in a reference center for the treatment of infectious diseases in the state of Minas Gerais . METHODS: Descriptive study including all children with probable diagnosis of meningitis between June/1999 and November/1999 . RESULTS: There were 210 cases of meningitis, and 111 (52.9%) were caused by bacteria . Actually, 52 were probable bacterial meningitis (with liquor alterations) and 59 were confirmed (with culture and/or antigen tests) . The main agents were: Haemophilus influenzae, Neisseria meningitidis and Streptococcus pneumoniae . The initial treatment for children aged between three months and five years consisted of ampicillin and chloramphenicol . Later, the antibiotic was changed to penicillin in cases of Neisseria meningitidis and Streptococcus pneumoniae . and to chloramphenicol in cases of Haemophilus influenzae . Extended spectrum antimicrobial agents were used on clinical or laboratory basis, but resistant microorganisms were not found in cultures . CONCLUSIONS: The epidemiology of meningitis should be continuous and should consider local data in order to guide antimicrobial therapy . The continuous monitoring of the prevalent agents in each institution and their resistance is fundamental to the selection of antimicrobial drugs, preserving the antimicrobial agents, and causing less interference with individual colonization, without contributing to the increasing resistance of the agents responsible for meningeal infections.

J Pediatr (Rio J), 2002 Nov-Dec, 78(6), 467 - 74
{Etiological profile of bacterial meningitis in children}; Mantese OC et al.; OBJECTIVE: To determine the etiologic profile and analyze some epidemiological aspects of children with bacterial meningitis admitted to a public teaching hospital . METHODS: A prospective study was conducted on children with clinical and laboratory diagnosis of bacterial meningitis, admitted to Hospital das Clinicas da Universidade Federal de Uberlandia, from January 1987 to January 2001 . Patients with meningitis associated with trauma, intracranial devices or malformations of the neural tube, and tuberculosis, were not included in the study . RESULTS: From a total of 415 children with bacterial meningitis, the etiologic agent was detected in 315 (75.9%): Haemophilus influenzae b in 54.2%, meningococci in 20.6%, pneumococci in 18.1% and other agents, in 6.9% . Previous antibiotic treatment, observed in 47.2% of the cases, led to a significant decrease in positive blood cultures (from 50.8% to 38.7%) and in cerebrospinal fluid cultures (from 71.7% to 57.6%) . Among children younger than 48 months Haemophilus influenzae b was predominant, particularly when compared to meningococci . The overall mortality was 10.1%, with a significant difference between the rates of pneumococcal (17.5%) and meningococcal meningitis (4.6%) . CONCLUSIONS: Children affected by Haemophilus influenzae b and by pneumococci were younger than those with meningitis caused by meningococci . The blood and/or cerebrospinal fluid culture remains an important laboratory tool for etiologic diagnosis, despite the negative impact caused by antibiotic previous treatment . The agents most commonly detected were Haemophilus influenzae b, meningococci and pneumococci . Bacterial meningitis continues to present an important mortality among children, particularly when caused by pneumococci.

J Bacteriol, 2003 Dec, 185(24), 7285 - 90
Differences in genetic and transcriptional organization of the glpTQ operons between Haemophilus influenzae type b and nontypeable strains; Song XM et al.; The glpTQ operon of Haemophilus influenzae type b (Hib) and nontypeable H . influenzae (NTHi) strains is highly conserved, except for a 1.4-kb glpTQ intergenic region that was found in most Hib strains . The presence of this intergenic region results in divergent glpTQ transcriptional profiles for Hib and NTHi where Hib strains appear to have evolved an alternative promoter for glpQ expression . Based on the intergenic region's low G+C content, we speculate that this DNA fragment was acquired by lateral transfer.

Microb Pathog, 2004 Jan, 36(1), 25 - 33
Reduced severity of middle ear infection caused by nontypeable Haemophilus influenzae lacking the hemoglobin/hemoglobin-haptoglobin binding proteins (Hgp) in a chinchilla model of otitis media; Morton DJ et al.; Since Haemophilus influenzae lacks enzymes necessary for synthesis of the porphyrin ring, it has an absolute growth requirement for a porphyrin source . This requirement can be satisfied in vitro by hemoglobin and hemoglobin complexed to haptoglobin . The products of the hgp genes mediate the utilization of heme from hemoglobin-haptoglobin . These genes are also involved in the use of heme from hemoglobin, although additional gene products independently mediate the acquisition of heme from this substrate . Different strains of H . influenzae possess one to four hgp genes . A nontypeable H . influenzae mutant lacking all the hgp genes was constructed and compared to the wild-type strain in a chinchilla (Chinchilla lanigera) model of otitis media . Compared to the wild-type strain, the hgp-deficient mutant exhibited a significantly delayed onset of detectable middle ear infection and significantly reduced duration of infection as assessed by both video otoscopy and tympanometry and as evidenced by viable bacterial counts in middle ear effusions . In addition, the maximum bacterial load in the middle ears of chinchillas infected with the mutant strain was significantly reduced when compared to the parent . These data indicate that the hemoglobin/hemoglobin-haptoglobin binding proteins are required for bacterial proliferation during H . influenzae-induced otitis media in chinchillas.

Arch Pediatr, 2003 Dec, 10(12), 1071 - 4
{Bilateral jugular thrombosis in Lemierre syndrome}; Benhayoun M et al.; An adolescent presented with a rhinosinusitis complicated with bilateral jugular veins and left superior ophthalmic vein thrombosis and respiratory distress with pulmonary hypertension . Blood culture was positive for Haemophilus influenzae and sinus puncture for Streptococcus constellatus . Evolution was under control after 1 week of appropriate antibiotherapy, antithrombotic and anti-inflammatory treatment . He had no neurologic, respiratory or ophthalmologic sequelae 6 months later . Despite lack of pharyngitis or isolation of anaerobic species on blood cultures, the picture was considered compatible with Lemierre syndrome . The risk for such a complication should be considered in cases of severe otorhinolaryngologic infection in young adults.

Int J Pediatr Otorhinolaryngol, 2003 Dec, 67(12), 1317 - 23
Microbiology of otorrhea in children with tympanostomy tubes: implications for therapy; Dohar J; OBJECTIVE: Based on the prevalence of children with tympanostomy tubes who develop otorrhea, an analysis was performed to identify specific prognostic indicators in this population to enable clinicians to determine the likelihood of specific pathogens and select the best empiric therapy . METHODS: Data from two multicenter clinical trials of ofloxacin otic solution 0.3% (OFLX) trials in pediatric patients 1-12 years of age were used to develop a statistical model to predict the likelihood of external auditory canal (EAC) or nasopharyngeal (NPG) pathogens . Data were available for 283 microbiologically evaluable patients . Potential indicators included subject age, season of enrollment, ear aspiration, cleaning, otorrhea, and granulation tissue . The model used a stepwise logistic regression analysis relating the occurrence of NPG or EAC pathogens to the potential prognostic indicators . RESULTS: Among the total study population, 42.8 and 61.5% had NPG and EAC pathogens, respectively; 10.6% had both . The most frequently isolated valid pathogens were Streptococcus pneumoniae and Haemophilus influenzae . Significant prognostic indicators for NPG pathogens were subject age, season of enrollment, and presence of ear odor . Although these indicators were similar for EAC and NPG pathogens, the correlation was reversed, i.e . older subjects had EAC pathogens, and younger ones (< 2 years) had NPG pathogens; EAC pathogens were associated with presence of ear odor and NPG pathogens, with absence of ear odor . CONCLUSIONS: A statistically and clinically valid model has been developed that has prognostic value for the clinician treating children with otorrhea and tympanostomy tubes and serves as an aid in the appropriate choice of empiric therapy.

Bone Marrow Transplant, 2004 Feb, 33(3), 337 - 46
Efficacy of donor vaccination before hematopoietic cell transplantation and recipient vaccination both before and early after transplantation; Storek J et al.; Allogeneic hematopoietic cell transplantation is followed by humoral immunodeficiency . We evaluated whether antibody levels can be improved by recipient vaccination on day -1 and 50 and whether the levels can be further improved by donor vaccination on day -20 . A total of 85 patients were randomized or assigned to one of the following strategies of immunization with Streptococcus pneumoniae polysaccharides, Haemophilus influenzae polysaccharide-protein conjugate, tetanus toxoid (protein recall antigen) and hepatitis B surface antigen (protein neo-antigen): (1) donor on day -20, recipient on days -1, +50 and +365 (D(-20)R(-1,50,365)); (2) donor nil, recipient on days -1, +50 and +365 (D(N)R(-1,50,365)); or (3) donor nil, recipient on day +365 (D(N)R(365)) . For H . influenzae and tetanus, IgG levels after grafting were the highest in the D(-20)R(-1,50,365) patients, intermediate in the D(N)R(-1,50,365) patients and the lowest in the D(N)R(365) patients . For S . pneumoniae and hepatitis B, antibody levels appeared to be similar in all three patient groups . The results suggest that for polysaccharide-protein conjugate antigens or protein recall antigens, recipient immunization on days -1 and 50 improves antibody levels and that donor vaccination on day -20 further improves the levels . In contrast, neither recipient immunization on days -1 and 50 nor donor immunization on day -20 appears to be efficacious for polysaccharide antigens and poorly immunogenic protein neo-antigens.

Infect Immun, 2003 Dec, 71(12), 7202 - 7
Genetic analysis of the capsule locus of Haemophilus influenzae serotype f; Satola SW et al.; A 19-kb DNA region containing genes involved in the biosynthesis of the capsule of Haemophilus influenzae serotype f (Hif) has been cloned and characterized . The Hif cap locus organization is typical of group II capsule biosynthetic loci found in other H . influenzae serotype b bacteria and other gram-negative bacteria . However, the Hif cap locus was not associated with an IS1016 element . Three new open reading frames, Fcs1, Fcs2, and Fcs3, were identified in the Hif capsule-specific region II . The chromosomal location of the Hif cap locus and the organization of the flanking sequences differed significantly from previously described division I H . influenzae serotypes.

Infect Immun, 2003 Dec, 71(12), 7178 - 82
Haemophilus ducreyi requires an intact flp gene cluster for virulence in humans; Spinola SM et al.; An intact Haemophilus ducreyi flp operon is essential for microcolony formation in vitro . tadA is the 9th of 15 genes in the operon and has homology to NTPases of type IV secretion systems . Fifteen human volunteers were experimentally infected with both H . ducreyi 35000HP and the tadA mutant, 35000HP.400 . Papules developed at similar rates at sites inoculated with the mutant and parent, while pustules formed at 36.4% of parent sites and at 0% of mutant sites (P = 0.001) . Compared to 35000HP, 35000HP.400 had only a modest but significant reduction in lesion scores in the temperature-dependent rabbit model of chancroid . These data suggest that proteins secreted by the flp locus are required for full expression of virulence by H . ducreyi in humans but have less of a role in virulence in an animal model of infection.

Infect Immun, 2003 Dec, 71(12), 6971 - 7
A humoral immune response confers protection against Haemophilus ducreyi infection; Cole LE et al.; Haemophilus ducreyi is the etiologic agent of the sexually transmitted genital ulcer disease chancroid . Neither naturally occurring chancroid nor experimental infection with H . ducreyi results in protective immunity . Likewise, a single inoculation of H . ducreyi does not protect pigs against subsequent infection . Accordingly, we used the swine model of chancroid infection to examine the impact of multiple inoculations on a host's immune response . After three successive inoculations with H . ducreyi, pigs developed a modestly protective immune response evidenced by the decreased recovery of viable bacteria from lesions . All lesions biopsied 2 days after the first and second inoculations contained viable H . ducreyi cells, yet only 55% of the lesions biopsied 2 days after the third inoculation did . Nearly 90% of the lesions biopsied 7 days after the first inoculation contained viable H . ducreyi cells, but this percentage dropped to only 16% after the third inoculation . Between the first and third inoculations, the average recovery of CFU from lesions decreased approximately 100-fold . The reduced recovery of bacteria corresponded directly with a fivefold increase in H . ducreyi-specific antibody titers and the emergence of bactericidal activity . These immune sera were protective when administered to naive pigs prior to challenge with H . ducreyi . These data suggest that pigs mount an effective humoral immune response to H . ducreyi after multiple exposures to the organism.

Infect Immun, 2003 Dec, 71(12), 6915 - 20
Th1-directing adjuvants increase the immunogenicity of oligosaccharide-protein conjugate vaccines related to Streptococcus pneumoniae type 3; Lefeber DJ et al.; Oligosaccharide (OS)-protein conjugates are promising candidate vaccines against encapsulated bacteria, such as Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pneumoniae . Although the effects of several variables such as OS chain length and protein carrier have been studied, little is known about the influence of adjuvants on the immunogenicity of OS-protein conjugates . In this study, a minimal protective trisaccharide epitope of Streptococcus pneumoniae type 3 conjugated to the cross-reacting material of diphtheria toxin was used for immunization of BALB/c mice in the presence of different adjuvants . Subsequently, half of the mice received a booster immunization with conjugate alone . Independent of the use and type of adjuvant, all mice produced long-lasting anti-polysaccharide type 3 (PS3) antibody levels, which provided full protection against challenge with pneumococcal type 3 bacteria . All adjuvants tested increased the anti-PS3 antibody levels and opsonic capacities as measured by an enzyme-linked immunosorbent assay and an in vitro phagocytosis assay . The use of QuilA or a combination of the adjuvants CpG and dimethyl dioctadecyl ammonium bromide resulted in the highest phagocytic capacities and the highest levels of Th1-related immunoglobulin G (IgG) subclasses . Phagocytic capacity correlated strongly with Th1-associated IgG2a and IgG2b levels, to a lesser extent with Th2-associated IgG1 levels, and weakly with thiocyanate elution as a measure of avidity . Thus, the improved immunogenicity of OS-protein conjugates was most pronounced for Th1-directing adjuvants.

Infect Immun, 2003 Dec, 71(12), 6884 - 91
Human antibodies specific for the high-molecular-weight adhesion proteins of nontypeable Haemophilus influenzae mediate opsonophagocytic activity; Winter LE et al.; The HMW1- and HMW2-like adhesion proteins of nontypeable Haemophilus influenzae are expressed by 75% of these strains, and antibodies directed against these proteins are protective in animal models of infection . The purpose of the present study was to define the functional activity of human antibodies specific for these proteins in an in vitro complement-dependent opsonophagocytic assay . Human promyelocytic cell line HL-60 served as the source of phagocytic cells, and a commercial preparation of intravenous immunoglobulin (IVIG) served as the source of human antibodies . High-molecular-weight (HMW) proteins were purified from four prototype nontypeable H . influenzae strains and used to prepare solid-phase affinity columns . IVIG was adsorbed on each column to remove strain-specific anti-HMW antibodies and to allow recovery of affinity-purified anti-HMW antibody fractions . Unadsorbed IVIG killed each of the prototype strains at titers of 1:80 to 1:320 . HMW-adsorbed sera demonstrated fourfold decreases in opsonophagocytic titer against the homologous strains compared to unadsorbed IVIG . Affinity-purified anti-HMW antibody preparations demonstrated opsonophagocytic titers of 1:20 to 1:80 against the respective homologous strains and opsonophagocytic titers as high as 1:80 against heterologous strains . None of the affinity-purified anti-HMW antibody preparations was opsonophagocytic for a representative nontypeable H . influenzae strain that did not express HMW1- or HMW2-like proteins . These data demonstrate that human antibodies specific for the HMW1/HMW2-like adhesion proteins of nontypeable H . influenzae are opsonophagocytic and that such antibodies recognize epitopes shared by the HMW proteins of unrelated nontypeable H . influenzae strains . These results argue for continued investigation of the HMW1/HMW2-like proteins as potential vaccine candidates for prevention of disease due to nontypeable H . influenzae.

Infect Immun, 2003 Dec, 71(12), 6784 - 92
Identification of dimethyl sulfoxide reductase in Actinobacillus pleuropneumoniae and its role in infection; Baltes N et al.; Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, is capable of persisting in oxygen-deprived surroundings, namely, tonsils and sequestered necrotic lung tissue . Utilization of alternative terminal electron acceptors in the absence of oxygen is a common strategy in bacteria under anaerobic growth conditions . In an experiment aimed at identification of genes expressed in vivo, the putative catalytic subunit DmsA of anaerobic dimethyl sulfoxide reductase was identified in an A . pleuropneumoniae serotype 7 strain . The 90-kDa protein exhibits 85% identity to the putative DmsA protein of Haemophilus influenzae, and its expression was found to be upregulated under anaerobic conditions . Analysis of the unfinished A . pleuropneumoniae genome sequence revealed putative open reading frames (ORFs) encoding DmsB and DmsC proteins situated downstream of the dmsA ORF . In order to investigate the role of the A . pleuropneumoniae DmsA protein in virulence, an isogenic deletion mutant, A . pleuropneumoniae DeltadmsA, was constructed and examined in an aerosol infection model . A . pleuropneumoniae DeltadmsA was attenuated in acute disease, which suggests that genes involved in oxidative metabolism under anaerobic conditions might contribute significantly to A . pleuropneumoniae virulence.

Antimicrob Agents Chemother, 2003 Dec, 47(12), 3831 - 9
Novel antibacterial class; Dandliker PJ et al.; We report the discovery and characterization of a novel ribosome inhibitor (NRI) class that exhibits selective and broad-spectrum antibacterial activity . Compounds in this class inhibit growth of many gram-positive and gram-negative bacteria, including the common respiratory pathogens Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, and Moraxella catarrhalis, and are nontoxic to human cell lines . The first NRI was discovered in a high-throughput screen designed to identify inhibitors of cell-free translation in extracts from S . pneumoniae . The chemical structure of the NRI class is related to antibacterial quinolones, but, interestingly, the differences in structure are sufficient to completely alter the biochemical and intracellular mechanisms of action . Expression array studies and analysis of NRI-resistant mutants confirm this difference in intracellular mechanism and provide evidence that the NRIs inhibit bacterial protein synthesis by inhibiting ribosomes . Furthermore, compounds in the NRI series appear to inhibit bacterial ribosomes by a new mechanism, because NRI-resistant strains are not cross-resistant to other ribosome inhibitors, such as macrolides, chloramphenicol, tetracycline, aminoglycosides, or oxazolidinones . The NRIs are a promising new antibacterial class with activity against all major drug-resistant respiratory pathogens.

Acta Paediatr, 2003 Oct, 92(10), 1163 - 9
Hand, foot and mouth disease in Singapore: a comparison of fatal and non-fatal cases; Chong CY et al.; AIM: An epidemic of hand, foot and mouth disease (HFMD) occurred in Singapore between September and November 2000 . During the epidemic, there were four HFMD-related deaths and after the epidemic, another three HFMD-related deaths . This study sought to determine the risk factors predictive of death from HFMD disease . METHODS: The risk factors for fatal HFMD were determined by comparing clinical and laboratory findings between fatal cases (n = 7) and non-fatal controls (n = 131) admitted between September 2000 and April 2001 . Enterovirus 71 positive fatal cases (n = 4) and non-fatal controls (n = 63) were also compared . RESULTS: In total, 138 HFMD cases with a mean age of 32 mo were studied . The majority of fatal cases died from interstitial pneumonitis, of whom three also had brainstem encephalitis . Of the 131 non-fatal cases, 3 had concomitant infections (respiratory syncytial virus bronchiolitis, right-sided pneumonia, Haemophilus influenzae type b meningitis), 2 had aseptic meningitis, and 1 each had transient drowsiness, intravenous immunoglobulin-related complications and transverse myelitis . By multivariate logistic regression analysis, atypical physical findings (p = 0.0006), raised total white cell count (p = 0.0128), vomiting (p = 0.0116) and absence of mouth ulcers (p = 0.043) were predictive of a fatal course . Although previous epidemics have described neurogenic pulmonary oedema as the main cause of death, the fatal cases in this study died mainly from interstitial pneumonitis alone or with myocarditis or encephalitis . CONCLUSION: Although HFMD is generally a benign disease, risk factors such as vomiting, absence of mouth ulcers, atypical presentation and raised total white cell count should alert the physician of a fatal course of illness.

Paediatr Drugs, 2003, 5 Suppl 1, 25 - 33
Acute bacterial rhinosinusitis in pediatric medicine: current issues in diagnosis and management; Anon JB; In children, acute bacterial rhinosinusitis is a common infection and although rare, carries a potential for serious, life threatening complications . Bacterial rhinosinusitis usually follows a viral infection or allergic rhinitis . Early, effective antibacterial therapy is essential to shorten the duration of infection and illness, to diminish mucosal damage, and to prevent contiguous infectious involvement of the orbit or central nervous system . Because the signs and symptoms of acute bacterial rhinosinusitis are similar to those of viral upper respiratory tract infection, establishing an accurate diagnosis in children poses a clinical challenge . Infection with Streptococcus pneumoniae accounts for 30-66% of episodes of acute bacterial rhinosinusitis in children . Other important pathogens include Haemophilus influenzae (20-30%) and Moraxella catarrhalis (12-28%) . In selecting initial antimicrobial therapy, priority should be given to drugs with activity against S . pneumoniae . The oral agents that currently offer the greatest activity against this pathogen include amoxicillin, amoxicillin-clavulanate, cefdinir, cefpodoxime proxetil, and cefuroxime axetil; all are considered appropriate for the initial treatment of acute bacterial rhinosinusitis in children . Amoxicillin is customarily used as first-line therapy for uncomplicated acute bacterial rhinosinusitis . For patients who are allergic to amoxicillin, second- or third-generation oral cephalosporins may be used as first-line therapy . Clarithromycin has been suggested as an alternative to amoxicillin or cephalosporins in beta-lactam allergic patients . Clindamycin may also be indicated as first-line treatment in patients who have culture-proven penicillin-resistant S . pneumoniae . If no clinical response occurs within 72 hours, the choice of a second-line antibiotic is governed by the drug's known antimicrobial efficacy, resistance patterns, dosing schedules, the potential for compliance, and knowledge of the patient's drug allergies . High-dose amoxicillin-clavulanate (90 mg/kg/d of the amoxicillin component) has been recommended for high-risk children (e.g . those in day care, and those who have recently received antibiotics) who show no improvement after treatment with the usual dose of amoxicillin (45 mg/kg/d) . Broad-spectrum, third-generation oral cephalosporins, such as cefdinir, should be considered as second-line agents when standard therapy has failed or when patients show hypersensitivity to penicillin . Intramuscular ceftriaxone may be appropriate for patients who fail on a second course of antibiotic treatment.

Paediatr Drugs, 2003, 5 Suppl 1, 1 - 12
Acute otitis media in pediatric medicine: current issues in epidemiology, diagnosis, and management; Leibovitz E; Acute otitis media (AOM) is not only the most common bacterial infection in children in the United States, it is also the most common indication for the prescription of antibiotics . Unfortunately, antibiotic resistance to pathogens (Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis) typically causative of AOM, continues to increase . More than 30% of the beta-lactamase producing H . influenzae are resistant to amoxicillin and virtually all strains of M . catarrhalis are beta-lactamase-positive . The emergence of multidrug-resistant strains, particularly S . pneumoniae, complicates the management of AOM and increases the risk for treatment failure . Because of growing resistance, the Centers for Disease Control and the American Academy of Pediatrics promote the judicious use of antibiotics in the treatment of AOM . Their recommendations emphasize the importance of distinguishing AOM from otitis media with effusion, minimizing the use of antibiotics, and discerning between first- and second-line antibiotics in the treatment of simple uncomplicated AOM versus non-responsive/recurrent AOM . Because spontaneous cure rates are lower in complicated AOM and AOM secondary to S . pneumoniae infection, antibiotic therapy remains an appropriate treatment option for most children with AOM . When amoxicillin, the treatment of choice in AOM, is not effective or not tolerated in children, the prescriber should consider an alternative that displays not only excellent antimicrobial activity against the suspected pathogens, but also characteristics, such as convenient dosing, tolerability, and palatability, that promote compliance and adherence in children . The cephalosporins offer an alternative to penicillins . Cephalosporins such as cefuroxime axetil (second-generation) and cefdinir and cefpodoxime proxetil (third-generation), offer a broad spectrum of activity and are approved for use in a convenient once- or twice-daily dosing schedule, thus increasing the likelihood of compliance with the full course of therapy . Cefdinir is a possible second-line alternative to amoxicillin for children with AOM, particularly among children who are likely to be noncompliant with a two- to three-times-daily dosing schedule, and those instances where there is a high likelihood for, or a known infection with an amoxicillin-resistant pathogen.

Presse Med, 2003 Sep 6, 32(28 Suppl), S12 - 4
{Antipneumococcal vaccines in sickle-cell anemia and asplenia}; Cohen R; TWO MEANS OF PREVENTION: Asplenic and sickle-cell anemia patients are susceptible to infections caused by polysaccharide-encapsulated bacteria, particularly pneumococci, Haemophilus influenzae b, and meningococci . Prevention of these frequent and serious infections depends on antibiotic prophylaxis and vaccination . ANTI-POLYSACCHARIDE PNEUMOCOCCI: Vaccination Pneumo 23 is a 23-valent vaccine corresponding to more than 90% of the pneumococci strains implicated in systemic infections . The efficacy of conjugated polysaccharide vaccines for the prevention of systemic infections, particularly when caused by antibiotic-resistant strains, can optimize the efficacy potential of antibiotic prophylaxis, the immense majority of strains not covered by the conjugated vaccine being susceptible to penicillin . CONJUGATED PNEUMOCOCCAL VACCINE: Prevenar is a seven-valence vaccine which induces a thymodependent antibody response . The efficacy of the conjugated vaccine against systemic infections, particularly antibiotic resistant strains, can have a potentializing effect on antibiotic prophylaxis, the vast majority of the strains not present in the conjugated vaccine being sensitive to penicillin . OTHER VACCINES: Finally, it should be recalled that conjugated antigroup C meningococcal vaccines are recommended for asplenic and sickle-cell anemia patients . In France, the anti-Haemophilus b vaccine is included in the vaccination calendar for all infants.

Presse Med, 2003 Sep 6, 32(28 Suppl), S5 - 9
{Asplenia and hyposplenism}; Beytout J et al.; FROM ASPLENIA TO SEVERE INFECTION: The most serious consequence of asplenia due to absence of the spleen, its resection or its functional failure is the risk of severe infection . RAPIDLY PROGRESSIVE SEPTICEMIA: It is estimated that the risk of death due to septicemia is 200 times higher in splectomized patients than in patients with a spleen . Death occurs within several days or even hours in most of the patients due to overwhelming post-splenectomy infection (OPSI) . The bacteria causing OPSI are predominantly pneumococci (50-80% of identified infections) as well as meningococci, Haemophilus sp, and other capsulated bacteria . IMPORTANCE OF AGE: The risk of infection is even greater if asplenia began early in life, either because of rapidly progressive congenital hemotological disease or splenectomy during infancy or early childhood . According to Holdsworth, infectious morbidity in subjects splenectomized before the age of 16 years is 4.4%, mortality reaches 2.2% . In adults, morbidity is 0.9% and mortality 0.8% . PREVENTION OF RECURRENCE: Furthermore, in an asplenic subject or in a patient with a chronic disease threatening the spleen, the development of infectious episodes is an expression of evolving immunodepression, calling for preventive measures against recurrence . Anti-pneumococcal vaccination and antibiotic prophylaxis using penicillin V considerably reduces the incidence of pneumococcal infection in splenectomized subjects.

Pediatr Res, 2004 Feb, 55(2), 347 - 56 Epub 2003 Nov 19.
Childhood vaccine development: an overview; Baker JP et al.; Vaccines against childhood diseases represent some of the most important applications of 20th-century pediatric research . This survey examines how the components of the current U.S . immunization schedule emerged in three phases during the course of the century . The first phase, after the development of bacterial culture techniques, witnessed numerous efforts in the early 1900s to develop bacterial vaccines . It proved most fruitful with respect to diphtheria, tetanus, and pertussis . The rise of viral tissue culture techniques in the 1950s brought about a second phase of innovation resulting in vaccines against polio, measles, mumps, rubella, and varicella . A third wave of innovation, still very much alive, has drawn on a variety of new technologies and led to vaccines against hepatitis B, Haemophilus influenzae type b, pneumococcus, and still other organisms . Although basic science research has thus been a primary factor shaping the history of vaccine development, the collaboration between the academic, private, and public sectors critical to its application has not always proceeded smoothly . The history of vaccine research and development has important implications for today, as a variety of factors threaten to fragment this network.

Int J Clin Pract, 2003 Oct, 57(8), 686 - 8
Nasopharyngeal carriage rate of Haemophilus influenzae in children aged 7-12 years in Turkey; Ayyildiz A et al.; This study was undertaken to determine the nasopharyngeal carriage prevalence of Haemophilus spp . in children aged 7-12 years in Erzurum, Turkey . Three hundred randomly selected students from three regions with different socioeconomic properties were included in the study . Nasopharyngeal swabs taken from the students were streaked into selective media for Haemophilus spp . Isolated colonies were identified by macroscopic and microscopic examination and their X and V factor needs determined . Those identified as H . influenzae were serotyped by specific capsule antisera . Haemophilus spp . were recovered from 51 (17.0%), nine (3.0%) of which were H . influenzae type b (Hib), 30 (10.0%) H . influenzae non-type b, and 12 (4.0%) H . parainfluenzae . Results were evaluated by several risk factors such as age, gender, number of siblings, siblings attending a nursery, family smoking habits and family economic status.

Bioorg Med Chem Lett, 2003 Dec 1, 13(23), 4209 - 12
New antibacterial tetrahydro-4(2H)-thiopyran and thiomorpholine S-oxide and S,S-dioxide phenyloxazolidinones; Singh U et al.; Combinatorial libraries of N-acylated 5-(S)-aminomethyloxazolidinone derivatives of S-oxide and S,S-dioxide tetrahydro-4(2H)-thiopyranyl and thiomorpholine phenyloxazolidinone series have been synthesized on a solid phase and evaluated for antimicrobial activity . Several novel potent leads have been identified, including orally active oxazolidinones with enhanced activity against respiratory tract infection pathogens Haemophilus influenzae and Moraxella catarrhalis.

Pediatr Pulmonol, 2003 Dec, 36(6), 469 - 74
Global burden of acute respiratory infections in children: implications for interventions; Mulholland K; Despite dramatic advances in human health that have occurred during the 20th century, the end of the century still sees many places in the world with high child mortality rates . This is made worse by increasing inequity, such that there are still many communities in the world in which over 30% of children die before their fifth birthday . Estimates of the global burden of childhood pneumonia are based on the assumption that there is a predictable relationship between the childhood mortality rate and the proportion of that mortality that is attributable to pneumonia . As most child deaths occur at home and can only be investigated by verbal autopsy techniques, these estimates are very crude and provide only a guide to the overall burden of pneumonia . Recent estimates from the World Health Organization suggest that 1.9 million children die as a result of acute respiratory infection (ARI), mainly pneumonia, each year . For a number of reasons, this is likely to be an underestimate . Estimates of the morbidity burden attributable to pneumonia are also very approximate, as studies have used different and nonstandardized definitions of pneumonia . These estimates were originally used to assist with planning of ARI intervention activities and for advocacy to draw attention to the problem of ARI . Recently, the introduction of new vaccines against Haemophilus influenzae type b (Hib) and Streptococcus pneumoniae (pneumococcus) raised the prospect of prevention of pneumonia by vaccination . For reasons outlined in this paper, great caution must be exercised before using existing pneumonia burden estimates to predict mortality savings that may accompany the introduction of these vaccines into developing countries .

J Bacteriol, 2003 Dec, 185(23), 6990 - 4
Slipped-strand mispairing can function as a phase variation mechanism in Escherichia coli; Torres-Cruz J et al.; Slipped-strand mispairing (SSM) has not been identified as a mechanism of phase variation in Escherichia coli . Using a reporter gene, we show that sequences that cause phase variation by SSM in Haemophilus influenzae also lead to phase variation when introduced onto the chromosome of E . coli, and the frequencies of switching are in the biologically relevant range . Thus, the absence of SSM-mediated phase variation in E . coli does not appear to be due to a mechanistic constraint.

Mol Microbiol, 2003 Nov, 50(3), 1017 - 30
A Neisserial autotransporter NalP modulating the processing of other autotransporters; van Ulsen P et al.; Autotransporters constitute a relatively simple secretion system in Gram-negative bacteria, depending for their translocation across the outer membrane only on a C-terminal translocator domain . We have studied a novel autotransporter serine protease, designated NalP, from Neisseria meningitidis strain H44/76, featuring a lipoprotein motif at the signal sequence cleavage site . Indeed, lipidation of NalP could be demonstrated, but the secreted 70 kDa domain of NalP lacked the lipid-moiety as a result of additional N-terminal processing . A nalP mutant showed a drastically altered profile of secreted proteins . Mass-spectrometric analysis of tryptic fragments identified the autotransporters IgA protease and App, a homologue of the adhesin Hap of Haemophilus influenzae, as the major secreted proteins . Two forms of both of these proteins were found in the culture supernatant of the wild-type strain, whereas only the lower molecular-weight forms predominated in the culture supernatant of the nalP mutant . The serine-protease active site of NalP was required for the modulation of the processing of these autotransporters . We propose that, apart from the autoproteolytic processing, NalP can process App and IgA protease and hypothesize that this function of NalP could contribute to the virulence of the organism.

Clin Microbiol Infect, 2003 Oct, 9(10), 1048 - 50
Haemophilus segnis: a rare cause of endocarditis; Somers CJ et al.; This report presents a case of endocarditis due to Haemophilus segnis, which represents a speciation difficulty for the routine laboratory . In this study, a molecular approach provided speciation, which was confirmed phenotypically by a reference laboratory . The use of molecular genotypic analysis is an additional strategy in the investigation of endocarditis . It has applications not only in isolate identification but also in primary detection of infection, particularly in patients whose blood is culture negative by conventional methodologies.

Clin Microbiol Infect, 2003 Aug, 9(8), 893 - 6
Antimicrobial activity of LB10827, a new orally administered cephalosporin, tested against Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae; Deshpande LM et al.; A new orally administered cephalosporin, LB10827, was compared to 16 other antimicrobial agents tested against Streptococcus pneumoniae (520 strains), Haemophilus influenzae (302 strains) and Moraxella catarrhalis (188 strains) by reference broth microdilution methods . LB10827 (MIC90, 0.12 mg/L; highest MIC, 0.5 mg/L) was 8-16-fold more potent than cefdinir, cefpodoxime or cefuroxime when tested against S . pneumoniae . All Gram-negative strains were inhibited at </= 0.5 mg/L LB10827, which is an activity equal or superior to that of currently available and tested oral beta-lactams . LB10827 appears to be a promising agent worthy of continued investigations both in vitro and in vivo.

APMIS, 2003 Oct, 111(10), 989 - 94
A mouse model for acute otitis media; Melhus A et al.; To induce acute otitis media in the mouse and to describe the clinical and bacteriological course of the infection, middle ears of BALB/c, Swiss-Webster and C57BL/6 mice were inoculated with Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis . Systemic and local changes were monitored by clinical observations, otomicroscopy, and analysis of bacterial samples from blood and middle ears . Agglutination of mouse erythrocytes by M . catarrhalis was also tested . Depending on bacterial strain, bacterial dose, and mouse strain three responses were identified: acute otitis media, otitis media with serous effusion, or no reaction . BALB/c mice were the most susceptible animals . On day 3, 76% of the BALB/c mice had developed middle ear infection, 50% had a positive middle ear culture, 56% were bacteremic, and 10% had succumbed to a disseminated infection . The local infections lasted approximately a week . Animals which survived recovered without permanent deterioration or otomicroscopically discernible changes . In no case did M . catarrhalis induce a culture-positive middle ear infection, possibly due to an inability to agglutinate the mouse erythrocytes . The mouse model can become a useful tool in studies of pneumococcal and H . influenzae-induced otitis media, but the bacterial dose has to be carefully titrated and adjusted to the chosen mouse strain.

Vaccine, 2003 Dec 12, 22(2), 287 - 92
Long-term persistence of immunity after immunisation with Haemophilus influenzae type b conjugate vaccine; Makela PH et al.; Although Haemophilus influenzae type b (Hib) conjugate vaccines, after licensure in 1987, are now recommended for world-wide use, the duration of protective immunity afforded by them is not known . We therefore assessed the immunogenity at 9-10 years of age in 37 children who had received the first Hib conjugate, PRP-D, in infancy (the Hib-conjugate group) and were now given a dose of Hib polysaccharide (PS) as a test vaccine . The anti-Hib PS antibodies (Hib-ab) were measured before and after this test vaccination, and the values compared to those in 37 control children who had not previously received any Hib vaccine and in 13 children who had received Hib PS vaccine in infancy (the Hib-PS group) . Prior to the test vaccination, the Hib-ab concentrations in the Hib-conjugate group were 3.6-fold higher than in the control group . After the test vaccination, the Hib-conjugate group had higher total Hib-ab concentrations, higher proportion of IgG and higher avidity of Hib-ab than the control or the Hib-PS group, suggesting persisting immunological memory in a Hib-c group . A mathematical model, including memory, predicted accurately the Hib-ab concentrations, which are maintained through anamnestic responses to intervening stimuli (Hib or cross-reacting bacteria).

Toxicol Lett, 2003 Dec 15, 146(1), 93 - 100
Effects of two pediatric vaccines on autoimmune diabetes in NOD female mice; Ravel G et al.; The induction or exacerbation of autoimmune diseases is a potential adverse effect of immunostimulating drugs . Vaccines have been suspected of such actions . Epidemiological studies, however, have so far failed to demonstrate any causal relationship between vaccination and autoimmune diseases, including insulin-dependent diabetes mellitus (IDDM) . In this study, autoimmune diabetes-prone non-obese diabetic (NOD) mice were treated with two multivalent diphtheria, tetanus, pertussis, poliomyelitis and haemophilus vaccines (diphtheria, tetanus, acellular pertussis, inactivated polio (DTaP-IVP) or DTaP-IVP/Haemophilus influenza type b (Hib)) intraperitoneally at each of 10, 12 and 14 weeks of age . Although non-statistically significant, the incidence of autoimmune diabetes was slightly reduced by the DTaP-IVP vaccine . Blood glucose levels were actually significantly reduced in the mice treated with the DTaP-IVP vaccine relative to the untreated control mice . A slight decrease in blood glucose levels amongst the mice given the DTaP-IVP/Hib vaccine was also noted . Therefore this study does not support previous claims that children's vaccination might be associated with acceleration or exacerbation of IDDM.

Pediatr Infect Dis J, 2003 Nov, 22(11), 949 - 56
Open label, multicenter study of gatifloxacin treatment of recurrent otitis media and acute otitis media treatment failure; Arguedas A et al.; BACKGROUND: Recurrent otitis media and treatment failures of acute infections are refractory to therapy . Newer fluoroquinolones have excellent activity against respiratory pathogens, but their use in children has been limited because of concerns about adverse effects . METHODS: This was an open label, multicenter trial in which patients with recurrent otitis media or acute otitis media (AOM) treatment failure were treated with 10 mg/kg gatifloxacin oral suspension once daily for 10 days . Before treatment a tympanocentesis or a swab of middle ear fluid was obtained . Nasopharyngeal swabs were obtained at baseline and at the end of therapy . Efficacy was evaluated 3 to 10 days after cessation of treatment and at 3 to 4 weeks . Safety monitoring included special attention to any sign or symptom suggestive of joint or bone abnormality . RESULTS: The study enrolled 254 patients 6 months to 7 years of age, with one-half (52%) of the patients having recurrent otitis media, 17% having AOM treatment failure and 28% having both . Cure was achieved posttreatment in 88% of 198 clinically evaluable patients, with similar outcomes for patients younger or older than 2 years of age . Of the 45 evaluable patients with Streptococcus pneumoniae, 38 (84%) were cured, including 25 of 28 with penicillin-nonsusceptible strains . Also cured were 89% of those with Haemophilus influenzae and those with Moraxella catarrhalis . No selection of resistance to gatifloxacin was detected among nasopharyngeal pathogens . Eighty-three percent of the children had sustained cure at the 4 weeks follow-up visit . Adverse events were primarily mild gastrointestinal, with no occurrences of arthropathy . CONCLUSION: Gatifloxacin is safe and effective for treatment of recurrent otitis media and AOM treatment failure in children.

Pediatr Infect Dis J, 2003 Nov, 22(11), 943 - 9
Bacteriologic and clinical efficacy of oral gatifloxacin for the treatment of recurrent/nonresponsive acute otitis media: an open label, noncomparative, double tympanocentesis study; Leibovitz E et al.; BACKGROUND: Gatifloxacin is an 8-methoxyfluoroquinolone with good activity against respiratory pathogens . OBJECTIVES: To document the bacteriologic and clinical efficacy of gatifloxacin in recurrent/nonresponsive acute otitis media (AOM) . METHODS: One hundred sixty patients 6 to 48 months of age with recurrent/nonresponsive AOM received gatifloxacin suspension (10 mg/kg once daily for 10 days) . Recurrent AOM was defined as > or =3 AOM episodes during the previous 6 months or > or =4 AOM episodes during the previous 12 months . Nonresponsive AOM was defined as AOM occurring < or =14 days after completing antibiotic treatment or not improving after > or =48 h of therapy . Middle ear fluid (MEF) obtained by tympanocentesis pretreatment (Day 1) and 3 to 5 days after initiation of treatment (Days 4 to 6) was cultured . Additional MEF cultures were obtained if clinical failure or recurrence of AOM occurred . Bacteriologic failure was defined by culture-positive MEF during treatment . Patients were followed until Days 22 to 28 . Susceptibility was determined by broth microdilution . RESULTS: One hundred twenty-eight (80%) patients completed treatment, and 32 discontinued the study prematurely (adverse events, 17; lost to follow-up, 10; consent withdrawal, 3; and laboratory abnormalities, 2) . From 89 patients (median age, 1 year; median number of prior AOM episodes, 4; range, 0 to 12), 121 pathogens were recovered: Haemophilus influenzae, 74 (61%); Streptococcus pneumoniae, 36 (30%); Moraxella catarrhalis, 9 (7%); and Streptococcus pyogenes, 2 (2%) . The 36 S . pneumoniae isolates were susceptible to gatifloxacin (MIC50 0.25 microg/ml); 26 of 36 (72%) were penicillin-nonsusceptible (15 fully resistant) . All 74 H . influenzae isolates were susceptible to gatifloxacin (MIC < or = 0.03 mg/ml) . Fourteen of 74 (19%) and 9 of 9 (100%) H . influenzae and M . catarrhalis isolates, respectively, produced beta-lactamase . Bacteriologic eradication was achieved for 118 of 121 (98%) pathogens: 74 of 74 H . influenzae; 34 of 36 (94%) S . pneumoniae; 9 of 9 M . catarrhalis; and 1 of 2 S . pyogenes . Clinical improvement/cure at end of treatment was seen in 103 of 114 (90%) clinically evaluable patients . Clinical recurrence of AOM after completion of therapy occurred in 31 patients . Of the 27 recurrent AOM cases in which tympanocentesis was performed, there were 16 (59%) new infections, 4 (15%) culture-negative results and only 7 (26%) true bacteriologic relapses . Adverse events were recorded in 21 of 160 (13%) patients: vomiting, 16; diarrhea, 3; maculopapular rash, 2 . No articular adverse events were recorded . CONCLUSION: Gatifloxacin is efficacious and safe for the treatment of recurrent/nonresponsive AOM.

J Antimicrob Chemother, 2003 Dec, 52(6), 961 - 6 Epub 2003 Nov 12.
Antibiotic resistance and clinical significance of Haemophilus influenzae type f; Campos J et al.; OBJECTIVES: Little is known about the antibiotic susceptibility and clinical significance of non-type b capsulated Haemophilus influenzae . We studied the antibiotic resistance patterns, plasmid carriage and clinical features of H . influenzae type f infections in Spain during 1996-2002 . PATIENTS AND METHODS: Forty-nine H . influenzae type f recovered from Spanish hospitals were analysed at a central laboratory where full microbiological and molecular epidemiological studies were carried out . Antimicrobial susceptibility testing was performed in accordance with NCCLS guidelines . RESULTS: Twelve strains (24.5%) were resistant to ampicillin and 22 (44.9%) to co-trimoxazole . Decreased susceptibility to clarithromycin, tetracycline, chloramphenicol and rifampicin was found in 16.3%, 12.2%, 14.3% and 2% of strains, respectively . Multidrug resistance was present in nine (18.4%) of the 49 isolates . The most prevalent resistance phenotype was ampicillin/tetracycline/co-trimoxazole/chloramphenicol, which was detected in five isolates . All six strains that were simultaneously resistant to ampicillin, tetracycline and chloramphenicol had conjugative plasmids . The main clinical diagnoses were pneumonia (32.6%), sepsis (18.4%) and meningitis (16.3%) . Thirty-two patients (65.3%) had previous underlying predisposing conditions, principally respiratory diseases (20.4%) . Twenty-one patients (42.8%) had impaired immunity . Thirty-seven (75.5%) patients were >14 years old, 12 (24.5%) were < or =14 years, and seven were < or =5 years . Most isolates were clonally related . CONCLUSIONS: A high prevalence of antibiotic resistance, including multiresistance, was detected in Spanish H . influenzae type f isolates . Carriage of large conjugative plasmids was strongly associated with antibiotic resistance . H . influenzae type f is mainly an opportunistic pathogen, although it may cause primary severe infections, such as meningitis in children.

J Antimicrob Chemother, 2003 Dec, 52(6), 1011 - 4 Epub 2003 Nov 12.
Molecular basis of rifampicin resistance in Haemophilus influenzae; Cruchaga S et al.; OBJECTIVES: To determine the molecular basis of rifampicin resistance in Haemophilus influenzae . METHODS: Mutations in the rifampicin-resistance determining region of the rpoB gene of H . influenzae were analysed by gene amplification and sequencing in 12 rifampicin resistant, one intermediate and four susceptible isolates . RESULTS: All clinical resistant isolates except one had at least one amino acid substitution in the beta-subunit of RNA polymerase . Eleven resistant isolates had amino acid changes at codons 513, 516, 518, 526 and 533 of cluster I, with the most common amino acid substitution being Asp-516-->Val . Only one resistant isolate also had a second mutation Asn-518-->Asp in cluster I; transformants obtained with DNA of this isolate also had both mutations . All the amino acid changes in cluster I were detected in isolates with a high level of rifampicin resistance (MIC > or =32 mg/L), except the Asp-516-->Ala mutation in a low-level resistant isolate (MIC 4 mg/L) . Only one serotype f isolate with an MIC of 2 mg/L had a mutation in cluster II . Cluster III presented no amino acid changes . In in vitro-generated high-level rifampicin-resistant mutants, only amino acid changes at codons 516 and 526 were seen, with new amino acid changes appearing at codon 526 of cluster I, while His-526-->Asn was associated with low-level resistance . CONCLUSIONS: Rifampicin resistance in H . influenzae is due to point mutations in the rpoB gene, and the resistance levels are dependent on both the location and the nature of amino acid substitution.

Int J Mol Med, 2003 Dec, 12(6), 1003 - 7
A novel splice variant of human gene GDPD1 is mainly expressed in human ovary and small intestine; Wu M et al.; Glycerophosphodiester phosphodiesterase (GDPD) is a glycerol metabolizing enzyme that has been previously identified in Escherichia coli, Haemophilus influenzae, Bacillus subtilis and Borrelia hermsii . It has been also reported that there is putative protein containing GDPD domain in Mus musculus, but not in Homo sapiens . SMART analysis showed that our predicted protein and three others of the same gene also comprise this domain . Because the gene has not been named, we termed it GDPD1 and accordingly termed our splice variant GDPD1_v1 after acquiring the agreement of HUGO . RT-PCR experiment revealed that our splice variant of the gene is mainly expressed in human ovary and small intestine, basically conforming to the result of serial analysis of gene expression.

Arch Pediatr Adolesc Med, 2003 Nov, 157(11), 1115 - 20
Effective pain reduction for multiple immunization injections in young infants; Reis EC et al.; BACKGROUND: Infants experience undue pain with multiple immunization injections . OBJECTIVE: To assess the effectiveness, feasibility, and parental acceptance of a simple combination pain reduction intervention for infants receiving multiple immunization injections . DESIGN: Randomized, controlled, clinical trial . SETTING: Academic hospital-based primary care center . PARTICIPANTS: Infants receiving their 2-month immunizations, consisting of 4 injections (diphtheria and tetanus toxoids and acellular pertussis vaccine, inactivated poliovirus vaccine, Haemophilus influenzae b conjugate and hepatitis B vaccine {Comvax}, and heptavalent pneumococcal conjugate vaccine {Prevnar}) . INTERVENTIONS: Subjects were randomly assigned to the intervention or control group for administration of 4 injections . The intervention group received sucrose and oral tactile stimulation (with a pacifier or a bottle) and were held by their parents during immunization . The control group did not receive these interventions (standard practice) . MAIN OUTCOME MEASURES: Blinded assessment of audiotaped crying, heart rate, parent preference for future use of the injection technique, and nurse-rated ease of vaccine administration . RESULTS: One hundred sixteen infants (mean +/- SD age, 9.5 +/- 2.0 weeks) participated . The median (25th-75th percentile range) first cry duration was 19.0 (5.8-62.8) seconds for the intervention group compared with 57.5 (31.0-81.5) seconds for the control group (P =.002) . Parents of the intervention group reported a stronger preference for future use of the injection procedure . For intervention vs control, the median (25th-75th percentile) parent preference visual analog scale score was 97.0 (82.0-100.0) vs 44.0 (5.0-77.2) (P<.001) (100 indicates definitely prefer) . Nurse-rated ease of vaccine administration was equivalent for both treatment groups . CONCLUSIONS: Combining sucrose, oral tactile stimulation, and parental holding was associated with significantly reduced crying in infants receiving multiple immunization injections . Parents stated a strong preference for future use of this method, and nurses found the intervention injection technique easy to apply.

Am J Otolaryngol, 2003 Nov-Dec, 24(6), 374 - 83
The rising incidence of adult acute epiglottitis and epiglottic abscess; Berger G et al.; OBJECTIVE: To examine preliminary observations that the incidence of adult acute epiglottitis has risen between 1986 and 2000 . MATERIALS AND METHODS: Demographics, annual and seasonal occurrences, clinical presentation, diagnostic procedures, treatment, airway management, and complications of 116 consecutive adult patients with laryngoscopically confirmed acute epiglottitis are presented . RESULTS: The mean annual incidence of acute epiglottitis per 100,000 adults significantly increased from 0.88 (from 1986-1990) to 2.1 (from 1991-1995) and to 3.1 (from 1996-2000) (P <.001) . This rise seems to be unrelated to Haemophilus influenzae type b infection but related to miscellaneous pathogenic bacteria . During these periods, the number of epiglottic abscesses increased concomitantly with the rise in the incidence of acute epiglottitis (from 4/14 episodes {29%}, to 8/38 {21%}, and to 16/66 {24%}, respectively), showing a relatively constant ratio between both phenomena (P =.843) . Twenty-five patients (21%) underwent airway intervention, 16 because of objective respiratory distress and 9 because of imminent respiratory obstruction . Stepwise logistic regression showed that drooling, diabetes mellitus, rapid onset of symptoms, and abscess formation were associated with airway obstruction . Diverse origins for the epiglottic abscess, either from coalescent epiglottic infection or from mucopyocele of the tongue base, are suggested . CONCLUSIONS: A rise in the incidence of acute epiglottitis and a concomitant rise in the number of epiglottic abscesses were established . Although the course of acute epiglottitis is often benign and can be safely treated with a conservative management approach, delayed airway obstruction may develop from a few hours to days after admission.

Int J Antimicrob Agents, 2003 Nov, 22(5), 548 - 50
Surveillance of Haemophilus influenzae among respiratory tract samples of Turkish children; Yagci A et al.; We conducted three prospective studies of Haemophilus influenzae in different groups of children . Pharyngeal swab samples were taken (i) . from 1382 healthy infants and children between 0 and 10 years of age (group 1), attending well child clinics (n=438), day care centres (n=440) and elementary schools (n=504), and (ii) . from 322 children aged 2-10 years (group 2), clinically diagnosed as having upper respiratory tract infection . Pharyngeal swab samples and sinus aspirates were obtained from 49 children between 2 and 9 years of age (group 3), clinically diagnosed as having sinusitis . H . influenzae was isolated in similar rates from 315 (22.7%) of children in group 1, 72 (22.3%) of children in group 2 and 12 (24.4%) of children in group 3 . Serotype b comprised 7, 5.2 and 2% of all H . influenzae isolates for group 1, 2 and 3, respectively . Production of beta-lactamase was detected in 1.0% of H . influenzae type b isolates in group 1, 1.2 and 6.1% of all isolates in group 2 and 3, respectively . There were no beta-lactamase negative ampicillin-resistant strains.

Int J Antimicrob Agents, 2003 Nov, 22(5), 508 - 15
Is it possible to achieve bacterial eradication in otitis media with effusion by empirical antibiotic high doses and concomitant administration of acetaminophen? A microbiological and pharmacological study in the gerbil model; Parra A et al.; The efficacy of amoxycillin-clavulanic acid (10 and 15 mg/kg amoxycillin) and erythromycin (20 and 50 mg/kg) was assessed in a gerbil model of otitis media with effusion induced by beta-lactamase-producing Haemophilus influenzae . Animals were divided into groups receiving acetaminophen concomitantly or not receiving it . Treatment started 2 h post-middle ear inoculation and continued t.i.d . for up to three doses . Middle ear samples were obtained on day 2 post-inoculation . Amoxycillin-clavulanic acid showed significantly higher efficacy than erythromycin, regardless of acetaminophen administration (P<0.05) . Amoxycillin-clavulanic acid middle ear concentrations exceeded the amoxycillin MIC (1/0.5 microg/ml) by 1.88-fold, whereas erythromycin concentrations were below MIC level of 4 microg/ml . Animals receiving acetaminophen showed significantly fewer polymorphonuclear cells and more Haemophilus organisms in the middle ear exudate.

Int J Antimicrob Agents, 2003 Nov, 22(5), 497 - 501
In vitro activity of telithromycin compared with macrolides and fluoroquinolones against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis; Kucukbasmaci O et al.; The in vitro activity of telithromycin was compared with erythromycin A, azithromycin, clarithromycin, moxifloxacin, gemifloxacin, levofloxacin, ciprofloxacin, penicillin G, ampicillin, cefuroxime and ceftriaxone against 336 consecutive strains (83 Streptococcus pneumoniae, 168 Haemophilus influenzae and 85 Moraxella catarrhalis) isolated from patients with community-acquired respiratory tract infections . Telithromycin (MIC(90), 0.008 mg/l) was the most active drug against S . pneumoniae . Telithromycin was also highly active against M . catarrhalis (MIC(90), 0.06 mg/l), but less active against H . influenzae (MIC(90), 4 mg/l).

Microbiology, 2003 Nov, 149(Pt 11), 3177 - 84
Genetic analysis of a plasmid encoding haemocin production in Haemophilus paragallinarum; Terry TD et al.; The full sequence of plasmid p250, isolated from Haemophilus paragallinarum strain HP250, has been obtained . The plasmid contains seven ORFs: a putative integrase, a putative replication protein (repB) and five ORFs similar to those from the haemocin (bacteriocin) hmcDCBAI operon from Haemophilus influenzae . Of 19 other non-plasmid-containing H . paragallinarum strains screened (11 serovar reference strains and 8 field isolates), 17 strains produced haemocin and were resistant to killing by strain HP250 . These strains, unlike strain HP250, have a chromosomally encoded haemocin operon . A number of other members of the family Pasteurellaceae were tested for haemocin sensitivity . Pasteurella avium, Pasteurella volantium and Pasteurella species A, all non-pathogenic bacteria found in the respiratory tract of chickens suffering from respiratory diseases, were sensitive to H . paragallinarum haemocin . However, amongst the pathogenic Pasteurellaceae, 50 % of P . multocida isolates and all five isolates of Pasteurella haemolytica tested were sensitive to the haemocin . Given the prevalence of haemocin production in H . paragallinarum strains, it may play a role in aiding colonization by inhibiting other Gram-negative bacteria that are associated with the respiratory tract in chickens . The origin of replication from plasmid p250 has been used to generate an Escherichia coli-H . paragallinarum shuttle vector which may be useful in genetically manipulating H . paragallinarum.

Microbiology, 2003 Nov, 149(Pt 11), 3165 - 75
The role of lex2 in lipopolysaccharide biosynthesis in Haemophilus influenzae strains RM7004 and RM153; Griffin R et al.; The locus lex2, comprising lex2A and lex2B, contributes to the phase-variable expression of lipopolysaccharide (LPS) of Haemophilus influenzae and was found to be present in 74 % of strains investigated . lex2A contains 5'-GCAA repeats which vary in number from 4 to 46 copies between strains . The locus was cloned from the serotype b strains RM7004 and RM153 and showed >99 % nucleotide sequence identity between these strains and the published lex2 sequence . Disruption of the lex2B gene in strain RM7004 resulted in truncation of some LPS glycoforms, shown by gel fractionation, with only one glycoform reacting with a digalactoside-specific monoclonal antibody, 4C4, compared with four LPS glycoforms in the more elongated LPS of the parent strain . Mass spectrometry and NMR analyses of LPS from the lex2B mutant revealed loss of the terminal digalactoside as well as the second beta-glucose extending from the first heptose of the inner core . The authors conclude that Lex2B is the beta-(1-4)-glucosyltransferase that adds the second beta-glucose to the first beta-glucose as part of the oligosaccharide extension from the first heptose of the LPS of strain RM7004 . Investigation of the expression of the lex2 locus indicated that the genes are co-transcribed and that both reading frames are required for addition of this second beta-glucose in a phase-variable manner.

Microbiology, 2003 Nov, 149(Pt 11), 3155 - 64
Foreign signal peptides can constitute a barrier to functional expression of periplasmic proteins in Haemophilus influenzae; Lam SL et al.; To study the periplasmic branch of iron (ferric ion) uptake systems in Gram-negative bacteria, genetic reconstitution experiments were initiated in Haemophilus influenzae involving exchange of the periplasmic iron-binding protein . The expression of many of the heterologous periplasmic ferric-binding proteins (FbpAs) was quite limited . Transformation experiments with the fbpA gene from Neisseria gonorrhoeae yielded two colony sizes with different phenotypic characteristics . The small colonies contained the intact N . gonorrhoeae fbpA gene and were deficient in utilization of transferrin iron . The large colonies contained hybrid H . influenzae/N . gonorrhoeae fbpA genes, were proficient in transferrin iron utilization and had enhanced levels of expression of FbpA . These hybrid genes included several that encoded the mature N . gonorrhoeae FbpA with the H . influenzae signal peptide . To more fully evaluate the effect of foreign signal peptides, a series of hybrid genes were prepared that exchanged the signal peptides from H . influenzae FbpA, N . gonorrhoeae FbpA and the TEM-1 beta-lactamase . The presence of the H . influenzae leader was required for functional expression of FbpAs and was shown to dramatically increase the level of beta-lactamase activity.

J Biomol Screen, 2002 Oct, 7(5), 429 - 32
A strategy for high-throughput assay development using leads derived from nuclear magnetic resonance-based screening; Hajduk PJ et al.; A strategy is described for the development of high-throughput screening assays against targets of unknown function that involves the use of nuclear magnetic resonance (NMR) spectroscopy . Using this approach, molecules that bind to the protein target are identified from an NMR-based screen of a library of substrates, cofactors, and other compounds that are known to bind to many proteins and enzymes . Once a ligand has been discovered, a fluorescent or radiolabeled analog of the ligand is synthesized that can be used in a high-throughput screen . The approach is illustrated in the development of a high-throughput screening assay against HI-0033, a conserved protein from Haemophilus influenzae whose function is currently unknown . Adenosine was found to bind to HI-0033 by NMR, and fluorescent analogs were rapidly identified that bound to HI-0033 in the submicromolar range . Using these fluorescent compounds, a fluorescence polarization assay was developed that is suitable for high-throughput screening and obtaining detailed structure-activity relationships for lead optimization.

Soc Hist Med, 2003 Apr, 16(1), 97 - 110
Scientific triumphalism and learning from facts: bacteriology and the "Spanish flu" challenge of 1918; Tognotti E; The devastating influenza pandemic known as 'Spanish flu', which killed at least 20 million people all over the world in 1918, was responsible for the first bitter blow inflicted on triumphant bacteriology, fortified by the series of resounding successes achieved in identifying the pathogenic agents of terrible diseases such as anthrax, cholera, tuberculosis, plague, and syphilis . Over-confidence and the idea, born of the Pasteur revolution, that every infectious disease was caused by a bacterium, had led the scientific community to accept the theory put forward by the German bacteriologist, Richard Pfeiffer, who, in 1892, believed he had identified the pathogenic influenza agent in a bacterium, Haemophilus influenzae . But, while the most appalling epidemic ever to sweep through the world since the 'Black Death' of the 1300s was still raging, the scientific community had to admit that influenza originated not from a microbe, but from a virus . This article aims to reconstruct the enlightening and little-known cultured/scientific events and issues of the dramatic crisis that bacteriology experienced in the autumn of 1918, with the consequent simultaneous collapse of both the 'Pfeiffer doctrine' on the microbial origin of influenza and the illusion of a world free of infectious diseases . This was an illusion destined to surface again at the end of the century and collapse with the advent of AIDS.

Am J Respir Crit Care Med, 2004 Feb 15, 169(4), 448 - 53 Epub 2003 Nov 03.
Strain-specific immune response to Haemophilus influenzae in chronic obstructive pulmonary disease; Sethi S et al.; Previous studies of immune response to Haemophilus influenzae after exacerbations of chronic obstructive pulmonary disease (COPD) have yielded contradictory results . Using homologous (infecting) strains and immunoassays to surface-exposed epitopes, we tested the hypothesis that adults with COPD make new antibodies to strain-specific, surface-exposed epitopes on H . influenzae after exacerbations . We collected clinical information, sputum, and serum monthly and during exacerbations from 81 patients with COPD over 56 months . Serum antibodies to H . influenzae after exacerbations associated with H . influenzae in sputum were detected with whole bacterial cell ELISA and bactericidal assays . An immune response to homologous H . influenzae occurred after 22 of 36 (61.1%) exacerbations with newly acquired strains compared with 7 of 33 (21.2%) exacerbations with preexisting strains (odds ratio {OR} = 4.4; 95%, 1.8 to 10.8; p = 0.001) . An absence of an immune response was strongly associated with complement sensitivity (OR = 0.03; 95% confidence interval, 0.003 to 0.22; p = 0.001) . New bactericidal antibodies developed after exacerbations were highly strain specific, showing bactericidal activity for only 11 of 90 (12.2%) heterologous strains . Development of an immune response to H . influenzae supports its role in causing exacerbations . The strain specificity of the immune response likely represents a mechanism of recurrent exacerbations.

Diagn Microbiol Infect Dis, 2003 Nov, 47(3), 515 - 25
Contemporary evaluation of the in vitro activity and spectrum of cefdinir compared with other orally administered antimicrobials tested against common respiratory tract pathogens (2000-2002); Sader HS et al.; Cefdinir is an oral cephalosporin approved by the Food and Drug Administration in 1997 for the treatment of acute exacerbation of chronic bronchitis, pharyngitis-tonsillitis, community-acquired pneumonia, acute maxillary sinusitis, and uncomplicated skin and skin structure infections in adults and adolescents, and acute otitis media, pharyngitis-tonsillitis, and uncomplicated skin and skin structure infections in children . Although cefdinir showed similar activity to other cephalosporins in the early studies, very limited data has been generated over the last decade . In this report, we summarize the contemporary in vitro activity and spectrum of cefdinir in comparison to numerous other orally administrated antimicrobials available for treatment of community-acquired respiratory infections . A total of 8,326 non-duplicate recent clinical isolates, including Haemophilus influenzae (3,438), Moraxella catarrhalis (1,688), and Streptococcus pneumoniae (3,200), were collected from 35 medical centers in North America during 2000 through 2002, and susceptibility tested by reference broth microdilution methods . Pneumococcal susceptibility patterns for beta-lactams and macrolides were also analyzed according to the year of isolation and the age group of the patients . Cefdinir had the greatest activity against H . influenzae among the cephalosporins tested with susceptibility rates of 97.1 to 99.0% . All of the agents tested had complete or near complete activity against M . catarrhalis . Against S . pneumoniae, cefdinir and other cephalosporins showed similar susceptibility patterns, but improved rates were observed in 2002 (78.5-79.4%) when compared to the previous monitored period (71.8-74.5%) . This increase in susceptibility was mainly because of a declining the occurrence of high-level penicillin resistance (MIC >/=2 microg/ml) across all age groups . Macrolide resistance also decreased among S . pneumoniae in 2002 when compared to 2000 through 2001; however, resistance to levofloxacin continued to increase from 0.9% in 2000 to 1.4% in 2002 . These results indicate a significant change in emerging beta-lactam resistance patterns (including cefdinir) with a decrease possibly influenced by greater pneumococcal vaccine use in children and the elderly . These rates of increased susceptibility could sustain and enhance the clinical activity of orally administered beta-lactams such as cefdinir.

Pediatrics, 2003 Nov, 112(5), 1049 - 53
Hearing loss at school age in survivors of bacterial meningitis: assessment, incidence, and prediction; Koomen I et al.; OBJECTIVES: To establish the incidence of sensorineural hearing loss in children who survived non-Haemophilus influenzae type B (Hib) bacterial meningitis, to highlight the actual percentage whose hearing was evaluated, and to develop a prediction rule to identify those who are at risk of hearing loss . METHODS: In 1999, we compiled a cohort of 628 school-aged children who were born between January 1986 and December 1994 and had survived non-Hib bacterial meningitis between January 1990 and December 1995 . Presence of sensorineural hearing loss (>25 dB) was determined, based on information from questionnaires and medical records . Potential risk factors for hearing loss were obtained from medical records; independent predictors were identified using multivariate logistic regression analysis, leading to the formulation of a prediction rule . RESULTS: The incidence of hearing loss was 7% . The hearing of 68% of the children was evaluated as part of their routine follow-up after bacterial meningitis, resulting in the detection of 75% of the cases of hearing loss . The remaining 25% were detected after this follow-up had ended . Using a prediction rule based on 5 factors-duration of symptoms before admission >2 days, absence of petechiae, cerebrospinal fluid glucose level <or=0.6 mmol/L, Streptococcus pneumoniae, and ataxia-62% of the postmeningitic children were selected as being at risk . All cases of hearing loss were in this at-risk group . CONCLUSIONS: Hearing loss can be predicted satisfactorily . When the hearing of children who are predicted to be at risk is tested as part of their routine follow-up, no children with hearing loss need be missed.

J Antimicrob Chemother, 2003 Dec, 52(6), 931 - 43 Epub 2003 Oct 29.
Antimicrobial susceptibility of lower respiratory tract pathogens in Great Britain and Ireland 1999-2001 related to demographic and geographical factors: the BSAC Respiratory Resistance Surveillance Programme; Reynolds R et al.; OBJECTIVE: The aim of this study was to assess the antimicrobial susceptibility of community-acquired lower respiratory pathogens in Great Britain and Ireland, and investigate its relationship with demographic and geographical factors using multiple logistic regression analysis . METHODS: A total of 1328 isolates of Streptococcus pneumoniae, 1894 Haemophilus influenzae and 845 Moraxella catarrhalis were collected from lower respiratory clinical specimens (primarily sputum) by 20 laboratories in Great Britain (England, Wales and Scotland) and Ireland (Northern Ireland and Eire) between 1999 and 2001 . RESULTS: Of 1154 S . pneumoniae from Great Britain, 92-100% were susceptible to beta-lactams (only 0.2% having penicillin MICs > or =2 mg/L), 89% were susceptible to erythromycin, 93% susceptible to tetracycline, and 94-100% intermediate or susceptible to fluoroquinolones . Susceptibility to agents other than fluoroquinolones was less frequent in the 174 isolates from Ireland: beta-lactam susceptibility was 68-99% (3.4% having penicillin MICs > or =2 mg/L), erythromycin susceptibility was 78% and tetracycline susceptibility was 82% . In multivariate analysis, susceptibility in S . pneumoniae was associated with country and patient age, being most common overall in the 20-49 years age group . Of 1894 H . influenzae, 15% produced beta-lactamase and 79-100% were susceptible to beta-lactams other than cefaclor . Ninety-six per cent were intermediate and 1% susceptible to erythromycin, 97% susceptible to tetracycline, and 89% susceptible to trimethoprim . Only one isolate showed resistance to ciprofloxacin . H . influenzae from sputum were more likely to be susceptible than isolates from other sources . Of 845 M . catarrhalis, 92% produced beta-lactamase and 9% were susceptible to ampicillin, >99% were susceptible to co-amoxiclav, cefotaxime, erythromycin and fluoroquinolones . CONCLUSIONS: Clinically relevant demographic factors predictive of susceptibility were country and patient age in S . pneumoniae, and specimen type (sputum/non-sputum) in H . influenzae . Susceptibility to most antimicrobials remains high in Ireland and very high in Great Britain.

J Antimicrob Chemother, 2003 Dec, 52(6), 925 - 30 Epub 2003 Oct 29.
Comparison of BSAC agar dilution and NCCLS broth microdilution MIC methods for in vitro susceptibility testing of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis: the BSAC Respiratory Resistance Surveillance Programme; Reynolds R et al.; OBJECTIVE: The aim of this study was to establish the degree of comparability between the NCCLS broth microdilution and BSAC agar dilution MIC methods of antimicrobial susceptibility testing . METHODS: Six hundred and sixty-one clinical isolates of Streptococcus pneumoniae, 936 Haemophilus influenzae and 421 Moraxella catarrhalis, collected in the winter of 1999-2000 by 20 laboratories in the UK and Eire from patients with presumed community-acquired lower respiratory infections, were tested by the two methods . MIC agreement was defined as excellent (good) if results were within +/- 1 doubling dilution for > or =90% (> or =80%) of isolates and within +/- 2 doubling dilutions for > or =95% . Isolates were categorized as susceptible, intermediate or resistant using the breakpoints appropriate to the testing method . RESULTS: MIC agreement was good or excellent in 27 of 36 organism-agent combinations . Agreement was less for M . catarrhalis than for other species, and lower in all three species for cefaclor and trimethoprim than for other antimicrobials . Discrepancies in categorization occurred only occasionally, and were generally explained by differences in breakpoints rather than in measured MICs . One exception was S . pneumoniae with penicillin . Despite excellent MIC agreement and identical breakpoints, 9% of these had minor discrepancies, mainly because 7% of isolates were found intermediate by the BSAC method but resistant by the NCCLS method . CONCLUSION: There is generally very good agreement between the MICs obtained by the BSAC agar dilution and NCCLS broth microdilution methods in this population of isolates, comparable to the level of agreement achieved between different laboratories using a single method . Breakpoint differences contribute to most of the discrepancies in susceptibility categorization.

J Antimicrob Chemother, 2003 Dec, 52(6), 1018 - 21 Epub 2003 Oct 29.
Contribution of beta-lactamase and PBP amino acid substitutions to amoxicillin/clavulanate resistance in beta-lactamase-positive, amoxicillin/clavulanate-resistant Haemophilus influenzae; Matic V et al.; The roles of beta-lactamase and alterations in penicillin-binding protein in the development of amoxicillin and amoxicillin/clavulanate resistance in two beta-lactamase-positive, amoxicillin/clavulanate-resistant (BLPACR) strains of Haemophilus influenzae were investigated . Seven beta-lactamase-negative, ampicillin-resistant (BLNAR) strains were also studied for comparison of their resistance mechanisms . All strains had been recovered from patients in Japan . The TEM type beta-lactamase of the two BLPACR strains had 100% homology with the amino acid sequences of published TEM-1 beta-lactamase, showing that amoxicillin/clavulanate resistance was not associated with mutations in this beta-lactamase . However, these strains, as well as the seven BLNAR strains, had multiple mutations in ftsI, which encodes penicillin binding protein 3 (PBP3) . The transformation of H . influenzae Rd strain with amplified ftsI genes from two BLPACR and two BLNAR strains enabled the selection of amoxicillin/clavulanate-resistant transformants with the same mutations as their parent strains . We concluded that amoxicillin/clavulanate resistance in the two BLPACR strains was due to changes in PBP3 . The possibility of the presence of an extended spectrum beta-lactamase was excluded in the BLPACR strains studied.

Cochrane Database Syst Rev . 2003;(4):CD001729.
Conjugate vaccines for preventing Haemophilus influenzae type b infections; Swingler G et al.; BACKGROUND: Haemophilus influenzae (H . influenzae) is an important cause of meningitis and pneumonia in children, causing an estimated three million cases of serious disease and hundreds of thousands of deaths annually worldwide . The introduction of H . influenzae type b (Hib) vaccines into routine immunisation schedules in developed countries has been followed by a rapid decline in disease occurrence, but vaccine cost is a significant barrier to use in developing countries . There is a need to determine the size of the effects of the vaccine, to enable cost-effectiveness comparisons with competing priorities in developing countries . OBJECTIVES: 1 . To determine the effects of conjugate Hib vaccine in preventing Hib disease or death in children under five years.2 . To determine any serious adverse outcomes . SEARCH STRATEGY: Searches of the Cochrane Central Register of Controlled Trials (CENTRAL) (issue 1, 2003); MEDLINE (January 1966 to April 2003), EMBASE (1990 to April 2003); and scanning of reference lists and contacting of authors of trial reports . Reports in all languages were considered . SELECTION CRITERIA: All randomised controlled trials or quasi-randomised trials of conjugate H . influenzae type b vaccines compared with placebo or no treatment in children who were followed until at least two years of age . DATA COLLECTION AND ANALYSIS: Two investigators independently selected eligible studies and extracted data . Differences were resolved by discussion . MAIN RESULTS: Five studies were included in the review, and four in meta-analyses . The overall quality of the trials was good.The relative risk for invasive Hib disease was 0.20 (95% confidence interval 0.07 to 0.54; random effects model), but there was statistically significant unexplained variation (heterogeneity) in the effects of the four trials in the meta-analysis (p = 0.002) . The size of the effects found in the trials did not appear to differ consistently with different vaccine types, the number of vaccine doses, age at first vaccination or use in developed vs developing countries, but the confidence intervals for the effect estimates were wide.Hib-related mortality data showed a non-significant trend towards benefit (relative risk was 0.29; 95% confidence interval 0.07 to 1.20; random effects model) . The relative risk for all cause mortality in the single trial from which data were available was 1.01 (95% confidence interval 0.38 to 2.67, random effects model).No serious adverse effects were reported in any of the trials, involving a total of 257,000 infants . REVIEWER'S CONCLUSIONS: Hib vaccine is safe and effective . The size of the effect could plausibly be anywhere between a 46% and 93% reduction in Hib invasive disease, before the effect of herd immunity is taken into account . In resource-poor settings, decisions to use the vaccine will depend on its cost, the local burden of Hib disease and competing priorities . Insufficient evidence from randomised controlled trials was identified of the effects of Hib conjugate vaccine on either Hib-specific or on all-cause mortality.

J Proteome Res, 2003 Sep-Oct, 2(5), 523 - 33
Proteome of Haemophilus ducreyi by 2-D SDS-PAGE and mass spectrometry: strain variation, virulence, and carbohydrate expression; Scheffler NK et al.; We have analyzed the proteome of several strains of Haemophilus ducreyi by two-dimensional gel electrophoresis (2-DE) and mass spectrometry . Over 100 spots were analyzed from the soluble and insoluble protein fractions from the prototype strain 35000HP and 122 distinct proteins were identified . Functions of approximately 80% of the 122 proteins were deduced by identification with close homologues of Haemophilus influenzae . Four additional wild type and three mutant strains were also analyzed that vary in their virulence and/or outer-membrane lipooligosaccharide structures . Overall, the 2-DE gel maps of the wild type and mutant strains were similar to strain 35000HP, suggesting little proteome diversity in relation to carbohydrate expression and/or virulence . An exception was the Kenyan strain 33921 which contained significant differences in its proteome 2-DE map and also synthesizes an unusual LOS with a trisaccharide branch structure . This African strain may represent a prototype of a second clonal group of H . ducreyi.

Microb Pathog, 2003 Dec, 35(6), 285 - 91
Caspase activation during Haemophilus somnus lipooligosaccharide-mediated apoptosis of bovine endothelial cells; Sylte MJ et al.; Vasculitis is commonly seen during systemic Haemophilus somnus infections . Although, the mechanism of vascular damage is not completely understood, in a previous report we demonstrated that H . somnus and its lipooligosaccharide (LOS) induced apoptosis in bovine pulmonary artery endothelial cells in vitro . In the present study, we investigated the role of caspase activation in LOS-mediated apoptosis of bovine endothelial cells . Exposure to H . somnus LOS induced caspase-3 activation and chromatin condensation in endothelial cells . These responses were blocked by the addition of a pan-caspase inhibitor (z-VAD-fmk) or capase-3 inhibitor (DEVD-fmk) . Incubation of endothelial cells with H . somnus LOS also induced activation of the initiator caspases, caspases-8 and 9, with the activity of the former increasing more rapidly than the latter . Addition of a caspase-8 inhibitor (IETD-fmk) significantly reduced LOS-mediated apoptosis, whereas, addition of a caspase-9 inhibitor (LEHD-fmk) had little effect . These data suggest that LOS-mediated activation of caspase-3 and apoptosis of endothelial cells is caspase-8 dependent.

Antimicrob Agents Chemother, 2003 Nov, 47(11), 3539 - 41
In vitro activities of garenoxacin (BMS-284756) against Haemophilus influenzae isolates with different fluoroquinolone susceptibilities; Perez-Vazquez M et al.; The in vitro activity of garenoxacin (BMS-284756) against 62 clinical Haemophilus influenzae isolates with different fluoroquinolone susceptibilities was determined by the microdilution susceptibility testing method and compared with the activities of other oral quinolones and nonquinolone oral antimicrobial agents . Cefixime presented the highest intrinsic activity (MIC at which 50% of the isolates tested were inhibited {MIC(50)}, 0.01 microg/ml), followed by garenoxacin, moxifloxacin, and ciprofloxacin (MIC(50), 0.06 microg/ml), levofloxacin (MIC(50), 0.12 microg/ml), cefuroxime (MIC(50), 1.0 microg/ml), and amoxicillin-clavulanate (MIC(50), 1.0/0.5 microg/ml), amoxicillin (MIC(50), 2 microg/ml), azithromycin (MIC(50), 4 microg/ml), and erythromycin (MIC(50), 8 microg/ml) . In strains with ciprofloxacin MICs of < or =0.06 microg/ml, ciprofloxacin and garenoxacin displayed similar MIC(50)s and MIC(90)s, one dilution lower than those of moxifloxacin and levofloxacin . For strains for which ciprofloxacin MICs were > or = 0.12 microg/ml, MIC(50)s were similar for the four quinolones tested, although garenoxacin presented the widest activity range (0.03 to 32 microg/ml) and the highest MIC at which 90% of the isolates tested were inhibited (16.0 microg/ml) . For strains without amino acid changes in the quinolone resistance determining region (QRDR) of GyrA and ParC, garenoxacin MICs were < or =0.03 microg/ml; with a single amino acid change in GyrA, garenoxacin MICs were 0.06 to 0.12 microg/ml; with one amino acid change each in GyrA and ParC, garenoxacin MICs were 0.5 to 2.0 micro g/ml; one amino acid change in ParC combined with two amino acid changes in GyrA increased the MICs to > or = 4 microg/ml for all assayed quinolones . We conclude that garenoxacin has excellent activity against H . influenzae, although progressive acquired resistance was observed by step-by-step mutation in the QRDR of gyrA and parC.

Acta Otolaryngol, 2003 Sep, 123(7), 873 - 8
Does the tonsillar surface flora differ in children with and without tonsillar disease?
Van Staaij BK, Van Den Akker EH, De Haas Van Dorsser EH, Fleer A, Hoes AW, Schilder AG.
OBJECTIVE: To investigate whether the tonsillar flora differ in children with and without adenotonsillar disease . MATERIAL AND METHODS: Tonsil surface swabs were taken from 218 children indicated for adenotonsillectomy because of moderate symptoms of recurrent tonsillopharyngitis or adenotonsillar hypertrophy (T+Ads group) . Control swabs were taken from 100 children without symptoms of adenotonsillar disease who visited the ophthalmology clinic . Potential respiratory pathogens were identified . RESULTS: Potential respiratory pathogens were found in 54% of the T+Ads group, compared to 41% of the control group (p = 0.04) . Haemophilus influenzae was the commonest pathogen in both groups, being found in 41% of the T+Ads group and 34% of the control group . Moraxella catarrhalis was found more often in the T+Ads group compared to the control group: 7% vs 0% (p = 0.004) . H . influenzae was found in 32% of the children with recurrent tonsillitis, compared to 48% of the children with symptoms of tonsillar hypertrophy (p = 0.03) . CONCLUSIONS: The prevalence of potential respiratory pathogens on the tonsillar surface of children with moderate symptoms of recurrent tonsillopharyngitis and/or tonsillar hypertrophy differs only slightly from that in children without symptoms of adenotonsillar disease . Variations in the microbial flora do not seem to play an essential role in the predisposition of these children to tonsillar disease.

Nature, 2003 Oct 23, 425(6960), 880 - 1
In search of form and function; Hoag H; On the genetics timeline, the first complete sequence of a free-living organism, that of the bacterium Haemophilus influenzae in 1995, was finished 130 years after Gregor Mendel published the results of his famous pea-breeding experiment . Since that sequence, others have followed thick and fast, including those of the nematode Caenorhabditis elegans, the fruitfly, humans and now the dog . But each sequence alone cannot provide researchers with much information about the organism . For that, they have to go through the painstaking process of knocking out genes, one at a time, and observing how these deletions affect biological function.

Infect Immun, 2003 Nov, 71(11), 6658 - 63
Differences in host susceptibility to disease progression in the human challenge model of Haemophilus ducreyi infection; Spinola SM et al.; With human volunteers inoculated at two sites with Haemophilus ducreyi, outcomes for a subject were not independent . In a reinfection trial, 2 of 11 previous pustule formers and 6 of 10 previous resolvers resolved all sites of infection . There was no correlation between serum bactericidal or phagocytic activity and outcome in the trial . These data indicate that different hosts are differentially susceptible to disease progression versus resolution in the model.

Infect Immun, 2003 Nov, 71(11), 6633 - 40
A CdtA-CdtC complex can block killing of HeLa cells by Haemophilus ducreyi cytolethal distending toxin; Deng K et al.; The cytolethal distending toxin (CDT) of Haemophilus ducreyi is comprised of the CdtA, CdtB, and CdtC proteins, with the CdtB protein having demonstrated enzymatic (i.e., DNase) activity . Using a single recombinant Escherichia coli strain with two plasmids individually containing the H . ducreyi cdtA and cdtC genes, we purified a noncovalent CdtA-CdtC complex . Incubation of this CdtA-CdtC complex with HeLa cells blocked killing of these cells by CDT holotoxin . Furthermore, the addition of purified recombinant CdtB to HeLa cells preincubated with the CdtA-CdtC complex resulted in the killing of these human epithelial cells.

South Med J, 2003 Oct, 96(10), 1008 - 17
Prevention and management of antibacterial resistance for primary care patients with respiratory tract infections; Iannini P; This review examines the problem of increasing antibacterial resistance among the pathogens commonly associated with community-acquired respiratory tract infections, particularly Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis . The increases in morbidity, mortality, and treatment cost associated with increased resistance to available antibiotics are challenging prescribers to find more effective therapeutic strategies . A MEDLINE search of the literature from 1966 to the present was performed to seek data relevant to the issue of resistance, especially the negative effects on patient outcomes and costs of therapy . Several observations and conclusions emerged . Data are lacking on local resistance patterns, broad-spectrum agents are overprescribed when narrower-spectrum choices would be more appropriate, a need exists for educational programs to encourage restricting drug use and changing prescribing habits, and there is a need for new antibiotic choices . The best antibiotic options are agents with a tailored spectrum of activity that are targeted at particular respiratory tract pathogens and have low potential to select for resistant organisms.

South Med J, 2003 Oct, 96(10), 974 - 85
Antimicrobial resistance among isolates of respiratory tract infection pathogens from the southern United States: data from the PROTEKT US surveillance program 2000/2001; Waites K et al.; BACKGROUND: PROTEKT US (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin in the United States) was established in 2000 to monitor antimicrobial resistance among respiratory tract pathogens across the United States . METHODS: During 2000 to 2001, 75 southern US centers collected 3,867 Streptococcus pneumoniae, 1,455 Streptococccus pyogenes and 1,042 Haemophilus influenzae . RESULTS: Overall, 46.1% of S . pneumoniae isolates were nonsusceptible to penicillin, 35.8% were resistant to erythromycin, and 0.5% were resistant to fluoroquinolones . Against S . pneumoniae the most active agents were telithromycin (99.7% susceptible), linezolid (99.8%) and the fluoroquinolones (levofloxacin 99.4%, gatifloxacin 99.5%) . The prevalence of erythromycin-resistant S . pyogenes isolates was 4.5% . Telithromycin, at concentration of < or = 1 mg/L, inhibited 99.9% of S . pyogenes . The prevalence of beta-lactamase positive H . influenzae was 26.2% . Telithromycin was active (MIC90 4 mg/L) against H . influenzae, irrespective of beta-lactamase production . CONCLUSION: The prevalence of penicillin and macrolide resistance among respiratory tract pathogens from the southern United States is high . Fluoroquinolone resistance is low . Telithromycin is highly active against respiratory tract pathogens with reduced susceptibility to beta-lactams, macrolides, and fluoroquinolones.

Pediatr Infect Dis J, 2003 Aug, 22(8 Suppl), S131 - 8
Macrolide resistance: an increasing concern for treatment failure in children; Jacobs MR et al.; BACKGROUND: Antimicrobial treatment of pediatric respiratory tract infections has evolved during the past 30 years as a result of antimicrobial resistance . The focus of antimicrobial therapy in these conditions has shifted from penicillins to other agents because of the dramatic increase in antimicrobial resistance among common respiratory pathogens, including Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis . It is important for clinicians to understand how resistance develops so that they can help prevent this phenomenon from occurring with other antimicrobials . METHODS: This article reviews the published literature on resistance to macrolide antimicrobials among common pediatric respiratory tract pathogens and clinical and bacteriologic outcomes of infections with these pathogens . RESULTS: Resistance among common pediatric respiratory tract pathogens to macrolides occurs through two main mechanisms, alteration of the target site and active efflux . Although resistance patterns vary by geographic region, the widespread use of macrolides has contributed to the emergence of both types of macrolide-resistant organisms . Conditions that favor the selection and proliferation of resistant strains include children with repeated, close contact who frequently receive antimicrobial treatment or prophylaxis, such as children who attend day care . Recent US surveillance data show that 20 to 30% of S . pneumoniae are resistant to macrolides, with approximately two-thirds of macrolide-resistant strains associated with an efflux mechanism and the remainder associated with a ribosomal methylase . Additionally, although less well-known, virtually all strains of H . influenzae have an intrinsic macrolide efflux pump . As resistance to macrolides has increased, clinical failures have resulted, and these agents are no longer considered appropriate for empiric first line antimicrobial therapy of acute otitis media and sinusitis unless patients are truly penicillin-allergic . Therefore, other antimicrobials are recommended for the empiric treatment of children with respiratory tract infections, including higher doses of amoxicillin and amoxicillin/clavulanate (90 mg/kg/day amoxicillin), cefuroxime axetil and intramuscular ceftriaxone . CONCLUSIONS: As resistance to macrolides increases and clinical failures in children become more common with this class of antimicrobials, judicious use of antimicrobials is needed . This includes limiting antimicrobial use for viral infections and using the most effective agents when antimicrobials are clinically indicated, such as higher doses of amoxicillin and amoxicillin/clavulanate . Application of these principles may prevent proliferation and further development of resistance.

Pediatr Infect Dis J, 2003 Aug, 22(8 Suppl), S120 - 30
Changes in treatment strategies for acute otitis media after full implementation of the pneumococcal seven valent conjugate vaccine; Harrison CJ; BACKGROUND: Administration of the pneumococcal seven valent conjugate vaccine (PCV-7) results in significant reductions in the incidence of invasive pneumococcal disease and nasopharyngeal colonization rates; however, the reduction in the overall incidence of acute otitis media (AOM) is minimal . Despite an ongoing shortage of PCV-7 in the United States, during which children are receiving fewer doses at longer intervals than the recommended dosing schedule, the incidence of invasive pneumococcal disease in young children has continued to decrease . PURPOSE: The purpose of this article is to review why the reported effect of PCV-7 on AOM infection rates does not match the reduction in invasive pneumococcal disease or the reduction in nasopharyngeal carriage of Streptococcus pneumoniae . METHODS: The potential effect of a fully implemented PCV-7 schedule on AOM infections is calculated based on serotype-specific AOM reduction rates observed in clinical studies . Clinical success rates of various antimicrobials used in the treatment of AOM in children fully immunized with PCV-7 also are calculated based on projected PCV-7-induced changes in pathogen profiles and documented antimicrobial resistance rates . Questions regarding potential serotype substitution in children with AOM but not invasive pneumococcal disease, the increased incidence of infection with Haemophilus influenzae in children immunized with PCV-7 and the possibility of herd immunity also will be addressed . RESULTS: Although the majority of penicillin-nonsusceptible S . pneumoniae are included in PCV-7, AOM caused by these strains will not completely disappear, especially in children 24 months of age and younger . CONCLUSIONS: Consequently, clinicians must continue to consider penicillin-nonsusceptible pneumococci when prescribing antimicrobials for AOM.

Pediatr Infect Dis J, 2003 Aug, 22(8 Suppl), S109 - 19
Worldwide trends in antimicrobial resistance among common respiratory tract pathogens in children; Jacobs MR; BACKGROUND: Respiratory tract infections among children are a common reason for health care provider visits and the primary reason for antimicrobial prescribing in this population . The increased prevalence of resistance among Streptococcus pneumoniae and Haemophilus influenzae pathogens poses a serious challenge in the successful treatment of respiratory tract infections caused by these pathogens . METHODS: This paper reviews worldwide trends in antimicrobial resistance among common respiratory tract pathogens, highlighting data obtained from the pediatric population where available . RESULTS: S . pneumoniae resistance to beta-lactams is mediated through alterations in the penicillin-binding proteins and macrolide resistance to acquisition of efflux or methylation genes . The mechanisms of resistance to the fluoroquinolones include target enzyme alterations via genetic mutations and transport out of the bacterial cell via an efflux pump . Beta-lactamase production is the primary mechanism of resistance to penicillins among H . influenzae isolates . Although S . pneumoniae with reduced susceptibility to penicillin was first documented > 30 years ago, resistance has increased at an alarming rate worldwide in the past decade . According to recent surveillance data, the worldwide prevalence of S . pneumoniae with reduced susceptibility to penicillin is 18.2% . Beta-lactamase production among H . influenzae ranges from approximately 4% in Russia to 26% in the United States and to 31% in France . The prevalence of beta-lactamase-negative, ampicillin-resistant H . influenzae remains very low (< 1%) worldwide, except in Japan, where the incidence is higher . In general, the highest rates of resistance are observed in isolates obtained from children, and risk factors for infection with a resistant pathogen include young age, the site of infection, day-care center attendance and recent antimicrobial use . CONCLUSIONS: Increased prevalence of antimicrobial resistance among respiratory tract pathogens isolated from children and adults is evident worldwide . Treatment of infections caused by S . pneumoniae and H . influenzae with older agents or ineffective dosing regimens may not eradicate infections and may contribute to the spread of resistance . These observations confirm the need for appropriate antimicrobial use to halt or at least limit the spread of resistance.

Gene, 2003 Oct 16, 316, 177 - 86
Inducible expression system and marker-linked mutagenesis approach for functional genomics of Haemophilus influenzae; Wong SM et al.; Complete bacterial genomic DNA sequences have allowed application of genome-scale strategies to identify essential genes . Efficient generation of conditional loss of function mutations provides a means of characterization of this class of genes . Promoter systems conferring tight regulation are particularly applicable to generating such mutations and we sought to apply this approach for the analysis of an essential gene of Haemophilus influenzae . Therefore, we developed the use of a D-xylose-inducible promoter for verification of an essential role in growth for yraM, which encodes a putative periplasmic lipoprotein, in both H . influenzae Rd and virulent type b strains . This promoter was sufficiently tightly regulated to generate conditionally viable strains by inducible expression of YraM . A second approach was used to further characterize YraM . Natural transformation and an ordered mutant collection spanning the H . influenzae genome provide the means to target any gene of interest for mutagenesis and temperature-sensitive (TS) mutant isolation . This strategy was applied to generate a conditionally lethal allele of yraM . The resulting TS mutation was directly mapped to a single amino acid substitution within a motif conserved in all putative YraM orthologs and this mutation was shown to be sufficient to confer the TS phenotype.

Infection, 2003 Aug, 31(4), 247 - 50
Cerebrospinal fluid glucose levels and sensorineural hearing loss in bacterial meningitis; Eisenhut M et al.; BACKGROUND: Low cerebrospinal fluid (CSF) glucose levels have been associated with sensorineural hearing loss (SNHL) in bacterial meningitis but systematic investigations are lacking . PATIENTS AND METHODS: A retrospective case control study was conducted comparing CSF glucose levels in 47 cases with and 145 controls without SNHL following bacterial meningitis . RESULTS: The mean CSF glucose level (standard deviation {SD}) was 1.3 mmol/l (1.2) in cases and 2.5 mmol/l (1.6) in controls (p < 0.001) . The difference between the groups was significant for Streptococcus pneumoniae and Haemophilus influenzae but not for Neisseria meningitidis infection . It was independent of the delay of treatment if this was greater than 12 h . This delay was significantly longer in cases compared to controls . CONCLUSION: In bacterial meningitis other than that caused by N . meningitidis and independent of the duration of symptoms prior to diagnosis, CSF glucose levels were significantly lower in patients developing a SNHL compared to controls.

Eur J Epidemiol, 2003, 18(9), 917 - 22
Risk of Haemophilus influenzae type b meningitis in Polish children varies directly with number of siblings: possible implications for vaccination strategies; Zielinski A et al.; BACKGROUND: In the absence of an effective vaccine, Haemophilus influenzae type b (Hib) meningitis has frequently been found to be the most common bacterial meningitis among children less than or equal to 5 years of age . This study was conducted prior to the introduction of Hib vaccine in Poland, and is the first case-control study of Hib meningitis in this country . METHODS: A 1:3 matched case-control study was nested within a population-based surveillance study of Hib meningitis in children less than or equal to 5 years old in Poland . Controls were matched on the case's age at onset of disease and immunization center . RESULTS: Having one or more siblings is a highly significant risk factor for Hib meningitis and, under the rare disease assumption, risk increases linearly with the increase in the number of siblings . The size of the living area of the home was not itself a significant risk factor for disease . Breast-feeding was not protective in contrast to previous studies . Childcare outside of the home was a significant risk factor for Hib meningitis especially among children greater than 16 months of age, whereas the effect of sibling number on Hib meningitis was much greater among the younger children . CONCLUSION: Risk of Hib meningitis approximately doubles for every unit increase in the number of siblings . Routine vaccination of Polish infants who have two or more siblings could potentially prevent half of the Hib meningitis cases.

Proc Natl Acad Sci U S A, 2003 Oct 28, 100(22), 12613 - 7 Epub 2003 Oct 14.
Capture of an intermediate in the catalytic cycle of L-aspartate-beta-semialdehyde dehydrogenase; Blanco J et al.; The structural analysis of an enzymatic reaction intermediate affords a unique opportunity to study a catalytic mechanism in extraordinary detail . Here we present the structure of a tetrahedral intermediate in the catalytic cycle of aspartate-beta-semialdehyde dehydrogenase (ASADH) from Haemophilus influenzae at 2.0-A resolution . ASADH is not found in humans, yet its catalytic activity is required for the biosynthesis of essential amino acids in plants and microorganisms . Diaminopimelic acid, also formed by this enzymatic pathway, is an integral component of bacterial cell walls, thus making ASADH an attractive target for the development of new antibiotics . This enzyme is able to capture the substrates aspartate-beta-semialdehyde and phosphate as an active complex that does not complete the catalytic cycle in the absence of NADP . A distinctive binding pocket in which the hemithioacetal oxygen of the bound substrate is stabilized by interaction with a backbone amide group dictates the R stereochemistry of the tetrahedral intermediate . This pocket, reminiscent of the oxyanion hole found in serine proteases, is completed through hydrogen bonding to the bound phosphate substrate.

Rev Prat, 2003 Sep 15, 53(13), 1442 - 5
{Influenza pneumonia}; Zeller V et al.; Two types of pneumonia are well recognized during influenza: primary viral pneumonia and secondary bacterial pneumonia . Primary viral pneumonia occurs after a typical onset of influenza with rapid progression of dyspnea and cough leading to acute respiratory distress syndrome . Treatment consists of respiratory assistance, but mortality is high . Secondary bacterial pneumonia occurs more frequently in the elderly and in patients with chronic pulmonary diseases . Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae are the most frequently isolated bacteria . After an initial phase of clinical improvement, manifestations of bacterial infection with pulmonary consolidation occur . The outcome is favorable with antibiotics but depends on the patient's underlying conditions.

Biochemistry, 2003 Oct 21, 42(41), 11908 - 14
Presence of ferric hydroxide clusters in mutants of Haemophilus influenzae ferric ion-binding protein A; Shouldice SR et al.; The periplasmic iron binding protein plays an essential role in the iron uptake pathway of Gram-negative pathogenic bacteria from the Pasteurellaceae and Neisseriaceae families and is critical for survival of these pathogens within the host . In this study, we report the crystal structures of two mutant forms of ferric ion-binding protein A (FbpA) from Haemophilus influenzae with bound multinuclear oxo-metal clusters . Crystals of site-directed mutants in the metal or anion binding ligands contain protein in the open conformation, and two mutant FbpAs, H9A and N175L, contain different cluster arrangements in the iron-binding pocket . The iron clusters are anchored by binding to the two tyrosine ligands (Tyr195 and Tyr196) positioned at the vertex of the iron-binding pocket but are not coordinated by the other metal binding ligands . Our results suggest that the metal clusters may have formed in situ, suggesting that the mutant FbpAs may serve as a simple model for protein-mediated mineralization.

Infection, 2003 Oct, 31(5), 324 - 6
Immunization with Haemophilus influenzae (Hib) conjugate vaccine following discharge from hospital; Olowokure B et al.; BACKGROUND: This study was undertaken to ascertain Haemophilus influenzae type b (Hib) conjugate vaccination status following discharge from hospital and to determine the time taken to vaccinate with Hib conjugate vaccine following hospital discharge of unvaccinated children . MATERIALS AND METHODS: Population-based cross-sectional survey involving children aged 3-59 months admitted to a hospital with laboratory confirmed invasive H . influenzae disease . RESULTS: Of 51 eligible children, 42 (82%) had not received Hib conjugate vaccine prior to admission . No children were immunized prior to discharge . Vaccination following discharge took an average of 110 days (range 3-441 days) . More than one quarter (28%) remained unvaccinated following discharge . If these children had been immunized, immunization coverage among this group of children would have increased significantly from 73% to 98% (p = 0.0017) . CONCLUSION: In the immediate post-Hib conjugate vaccine period, delayed immunization after discharge was identified . This reduces the individual and community benefits of immunization.

Braz J Infect Dis, 2003 Oct, 7(5), 315 - 24
Progress towards meningitis prevention in the conjugate vaccines era; Laval CA et al.; Acute bacterial meningitis is an important cause of morbidity and mortality among children less than five years old, Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis being the most important agents of bacterial meningitis in developing countries . The development of the conjugate vaccines in the beginning of the 90's, especially type b H . influenzae (Hib), and more recently the heptavalent pneumococcal and the serogroup C meningococcal vaccines, have contributed directly to changes in the epidemiological profile of these invasive diseases (direct effect) and of their carriage status (indirect effect) . We review the impact of the Hib conjugate vaccine in Latin American countries, where this vaccine has been implemented, and the potential of pneumococcal and meningococcal conjugate vaccines for the reduction of meningitis worldwide . We also address constraints for the development and delivery of these vaccines and review new candidate state-of-the-art vaccines . The greatest challenge, undoubtedly, is to implement these vaccines worldwide, especially in the developing regions.

Pediatr Infect Dis J, 2003 Oct, 22(10), 883 - 8
Double blind, randomized, placebo-controlled study of dexamethasone therapy for hematogenous septic arthritis in children; Odio CM et al.; BACKGROUND: Septic arthritis is associated with residual dysfunction in 10 to 25% of affected children . Concentrations of cytokines detected in synovial fluid of children with bacterial arthritis correlate with the severity of inflammation . Treatment with dexamethasone decreased cartilage degradation in experimental Haemophilus influenzae b and Staphylococcus aureus arthritis . ENDPOINTS: To decrease the number of patients with residual dysfunction of the affected joint at the end of therapy and at 6 and 12 months and to speed clinical recovery by the administration of dexamethasone . METHODS: In a double blind manner we randomly selected 123 children with suspected hematogenous bacterial arthritis to receive dexamethasone or saline for 4 days . Antibiotic therapy was tailored according to age and the recovered pathogen . RESULTS: Of the 123 children enrolled, 61 were assigned to the dexamethasone group and 62 to the placebo group . Only 50 and 50 patients in each group were evaluable . The 2 groups of patients were comparable with respect to age, sex, duration of symptoms, pathogen, affected joint and therapeutic and diagnostic procedures . Staphylococcus aureus accounted for 67% of the isolates, Haemophilus influenzae type b for 13% and Streptococcus pneumoniae for 9% . Dexamethasone therapy reduced residual dysfunction at the end of therapy, P = 0.000068; at 6 months, P = 0.00007; and at 12 months, P = 0.00053 of follow-up and shortened the duration of symptoms (P = 0.001) during the acute phase . The 26% incidence of residual dysfunction in the control patients was similar to the 25% found in other series . CONCLUSIONS: A short course of dexamethasone reduced residual joint dysfunction and shortened significantly the duration of symptoms in children with documented hematogenous septic arthritis . These results suggest that a 4-day course of low dose dexamethasone given early benefits children with hematogenous septic arthritis.

Pediatr Infect Dis J, 2003 Oct, 22(10), 878 - 82
Acute mastoiditis in Southern Israel: a twelve year retrospective study (1990 through 2001); Katz A et al.; BACKGROUND: Acute mastoiditis is a serious complication of acute otitis media (AOM) and has been increasingly reported in the last decade . OBJECTIVES: To report the experience accumulated with acute mastoiditis at the Soroka University Medical Center, Beer-Sheva, Israel, in a period of increasing antimicrobial resistance with Streptococcus pneumoniae . PATIENTS AND METHODS: We reviewed the records of all children with acute mastoiditis hospitalized from 1990 through 2001 . Acute mastoiditis was diagnosed when one or more of the physical signs of mastoiditis (swelling, erythema, tenderness of the retroauricular area and anteroinferior displacement of the auricle) were diagnosed in the presence of concomitant or recent (< or =4 weeks) AOM . RESULTS: One hundred sixteen episodes of acute mastoiditis occurred in 101 children age 2 months to 14 years (median, 25 months; 19% <1 year old) . The average yearly incidence was 6.1 cases per 100 000 population <14 years old, with a significant increase in the number of cases during the study period . Acute mastoiditis was the first evidence of AOM in 10 (10%) patients . Fever >38 degrees C and >15,000 WBC/mm3 were present in 67 and 43% of cases, respectively . Irritability, retroauricular swelling, redness and protrusion of the auricle occurred more commonly in patients <3 years old (79, 90, 84 and 76% vs . 28, 42, 45 and 30%, respectively, in patients > or =3 years old; P < 0.002) . Computed tomography scans were performed in 54 of 116 (47%) cases and revealed bone destruction in 38 (70%) . Periosteal abscess and lateral sinus vein thrombosis were diagnosed in 8 and 2 patients, respectively . Simple mastoidectomy was done in 32 of 116 (28%) cases, after no response to intravenous antibiotics was observed; ventilation tubes were inserted in 12 patients . Cultures were obtained at admission in 83 (72%) episodes (71 by tympanocentesis and 12 from ear discharges) . Overall 43 pathogens were isolated: 34 at admission; 14 at surgery; and 5 at both occasions . The most commonly isolated organisms were S . pneumoniae (14 of 43, 33%; 4 penicillin-nonsusceptible), Streptococcus pyogenes (11 of 43, 26%), nontypable Haemophilus influenzae (6 of 43, 14%), Pseudomonas aeruginosa (5 of 43, 12%) and Escherichia coli (4 of 43, 9%) . All S . pneumoniae organisms were isolated between 1996 and 2001 . CONCLUSIONS: (1) The incidence of acute mastoiditis in children in Southern Israel is greater than that reported in the literature; (2) a significant increase in the number of cases was recorded during the study period; (3) the clinical picture of acute mastoiditis was more severe in infants and young children; (4) the pathogen distribution in acute mastoiditis differs from that of AOM with significantly higher rates of S . pyogenes and lower rates of nontypable H . influenzae recovery; (5) penicillin-nonsusceptible S . pneumoniae played only a minor role in the etiology of acute mastoiditis in Southern Israel.

Int J Pediatr Otorhinolaryngol, 2003 Oct, 67(10), 1047 - 53
Microbial dynamics of purulent nasopharyngitis in children; Brook I; This review presents the microbiological dynamic and therapeutic options in the management of purulent nasopharyngitis (NPT) . The nasopharynx (NP) of healthy children is generally colonized by relatively non-pathogenic aerobic and anaerobic organisms, some of, which possess the ability to interfere with the growth of potential pathogens . Conversely, carriage of potential respiratory aerobic pathogen such as Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, as well as some anaerobic bacteria (Peptostreptococcus, Fusobacterium and Prevotella spp.) increases during purulent NPT . The development of purulent NPT in children is associated with the pre-existing colonization by potential pathogens and the absence of interfering organisms in the NP . Controversy exists regarding the management of NPT as no conclusive evidence exists to date that the administration of antimicrobials will shorten the illness.

Arq Gastroenterol, 2003 Jan-Mar, 40(1), 47 - 54 Epub 2003 Oct 06.
{Overwhelming postsplenectomy infection}; Marques RG et al.; BACKGROUND: Splenectomy performed at any age and for any reason increases the risk for death due to overwhelming infection . AIM: To evaluate definition, etiology, incidence, risk factors and prophylaxis of overwhelming postsplenectomy infection, as well as the methods related to splenic tissue preservation when total splenectomy is necessary . METHOD: Bibliographic review . RESULTS: The etiological agents more frequently found are Streptococcus pneumoniae, Haemophilus influenza and type B, and Neisseria meningitidis . Other bacteria like Escherichia coli, Streptococcus b-hemolytic, Staphylococcus aureus and Pseudomonas sp represent a significant risk as well . In addition, a great variety of agents including other enteric Gram-negative microorganisms and non-bacteria pathogens can also be sporadically identified . The prophylaxis is based on three main aspects: patient's education, immune prophylaxis and chemical prophylaxis . However these are not enough to prevent the higher risk of developing sepsis . When total splenectomy is unavoidable, heterotopic splenic autotransplantation seems to be the only alternative for splenic tissue preservation . According to clinical and experimental studies, the splenic autotransplanted tissue present a similar structure to a normal spleen and preserve the splenic immune function . CONCLUSIONS: The high risk of overwhelming postsplenectomy infection, reduced the indication for total splenectomy, in trauma and several diseases . Prophylactic methods have been developed to minimize the effects of the sepsis . Several researches have been done to determine the immunocompetence of autogenous splenic grafts in response to bacteria invasion.

Adv Exp Med Biol, 2001, 491, 515 - 24
Structure and functional genomics of lipopolysaccharide expression in Haemophilus influenzae; Richards JC et al.; The involvement of genes in the lic loci in H . influenzae LPS expression has been known for some time . However, it was not until recently that it was shown that the lic1 locus contains genes required for phase variable expression of phosphocholine substituents, while genes in the lic2 locus and lgtC are required for expression of the globoside trisaccharide, alpha-D-Galp-(1 --> 4)-beta-D-Galp-(1 --> 4)-beta-D-Glcp (i.e., the pK blood group epitope) . The availability of the complete sequence of the H . influenzae strain Rd genome has facilitated significant progress in understanding the role of these and other genes in the expression and biosynthesis of LPS . We have employed a comparative structural fingerprinting strategy to establish the structural relationships among LPS from H . influenzae mutant strains in which putative biosynthesis genes were inactivated . Using this functional genomics approach, we have gained considerable insight into the genetic basis for intra-strain and strain-to-strain variation in epitope expression.

Adv Exp Med Biol, 2001, 491, 453 - 71
Bacterial polysaccharides as vaccines--immunity and chemical characterization; Lee CJ et al.; Studies on protective immunity and biochemical characterization of bacterial capsular polysaccharides have led to significant contributions to understanding of the mechanisms of infectious diseases and development of effective vaccines . Immunity to encapsulated bacteria is related to antibody response to polysaccharide (PS) antigen, interactions with T- and B-lymphocytes, and host defense mechanisms . Meningococcal, pneumococcal and Salmonella vi PSs and Haemophilus type b PS-protein conjugate vaccines have been licensed and provided effective immunity for prevention of these bacterial infections . Capsular PSs are cell-surface polymers consisting of oligosaccharide repeating units . Many PSs are highly polar and hydrophilic and interfere with cell-to-cell interactions with phagocytes . Most pneumococcal PSs are negatively charged and possess acidic components such as D-glucuronic acid and phosphate in phosphodiester bonds . Extensive immunologic cross-reactivity has been observed among bacterial capsular PSs . In infants the antibody responses to most capsular PSs are generally poor . Enhanced immunogenicity of PS antigens can be achieved through PS-protein conjugate vaccines, immunization during a critical period of perinatal development and effective antigen delivery system.

J Med Microbiol, 2003 Nov, 52(Pt 11), 1005 - 14
Comparison of usefulness of randomly amplified polymorphic DNA and amplified-fragment length polymorphism techniques in epidemiological studies on nasopharyngeal carriage of non-typable Haemophilus influenzae; Augustynowicz E et al.; Randomly amplified polymorphic DNA (RAPD) and automated amplified-fragment length polymorphism (AFLP) techniques with fluorescently labelled primers were used to type non-serotypable Haemophilus influenzae (NTHI) isolates . Eighty-seven isolates from healthy children attending day-care centres or living at orphanages in southern Poland were investigated . Through comparison of the AFLP data with RAPD analysis, it has been concluded that the discriminatory power of AFLP for NTHI typing is higher than RAPD . Generally, the NTHI isolates analysed were highly heterogeneous, as detected with a HindIII/TaqI AFLP genotyping scheme on intra/inter similarity levels of 94 and 96 % using Pearson's correlation coefficient . The range of similarity values found for isolates from children permanently residing at a particular day-care centre was much wider than that for isolates from orphanages . AFLP can efficiently access NTHI strain diversity and can monitor their turn-over for comparative typing in local and inter-local epidemiological investigations.

J Biol Chem, 2003 Dec 26, 278(52), 52857 - 64 Epub 2003 Oct 06.
An isolated class II aminoacyl-tRNA synthetase insertion domain is functional in amino acid editing; Wong FC et al.; Aminoacyl-tRNA synthetases are responsible for activating specific amino acids and transferring them onto cognate tRNA molecules . Due to the similarity in many amino acid side chains, certain synthetases misactivate non-cognate amino acids to an extent that would be detrimental to protein synthesis if left uncorrected . To ensure accurate translation of the genetic code, some synthetases therefore utilize editing mechanisms to hydrolyze non-cognate products . Previously class II Escherichia coli proline-tRNA synthetase (ProRS) was shown to exhibit pre- and post-transfer editing activity, hydrolyzing a misactivated alanine-adenylate (Ala-AMP) and a mischarged Ala-tRNAPro variant, respectively . Residues critical for the editing activity (Asp-350 and Lys-279) are found in a novel insertion domain (INS) positioned between motifs 2 and 3 of the class defining aminoacylation active site . In this work, we present further evidence that INS is responsible for editing in ProRS . We deleted the INS from wild-type E . coli ProRS to yield DeltaINS-ProRS . While DeltaINS-ProRS was still capable of misactivating alanine, the truncated construct was defective in hydrolyzing non-cognate Ala-AMP . When the INS domain was cloned and expressed as an independent protein, it was capable of deacylating a mischarged Ala-microhelixPro variant . Similar to full-length ProRS, post-transfer editing was abolished in a K279A mutant INS . We also show that YbaK, a protein of unknown function from Haemophilus influenzae with high sequence homology to the prokaryotic INS domain, was capable of deacylating Ala-tRNAPro and Ala-microhelixPro variants but not cognate Pro-tRNAPro . Thus, we demonstrate for the first time that an independently folded class II synthetase editing domain and a previously identified homolog can catalyze a hydrolytic editing reaction.

Pediatrics, 2003 Oct, 112(4), 993 - 6
Increasing immunization coverage . American Academy of Pediatrics Committee on Community Health Services . American Academy of Pediatrics Committee on Practice and Ambulatory Medicine; Wood DL; American Academy of Pediatrics Committee on Community Health Services; American Academy of Pediatrics Committee on Practice and Ambulatory Medicine; Despite many recent advances in vaccine delivery, the goal for universal immunization set in 1977 has not been reached . In 2001, only 77.2% of US toddlers 19 to 35 months of age had received their basic immunization series of 4 doses of diphtheria and tetanus toxoids and acellular pertussis (DTaP) vaccine, 3 doses of inactivated poliovirus vaccine, 1 dose of measles-mumps-rubella (MMR) vaccine, and 3 doses of Haemophilus influenzae type b (Hib) vaccine . Children who are members of a racial or ethnic minority, who are poor, or who live in inner-city or rural areas have lower immunization rates than do children in the general population . Additional challenges to vaccine delivery include the introduction of new childhood vaccines, ensuring a dependable supply of vaccines, bolstering public confidence in vaccine safety, and sufficient compensation for vaccine administration . Recent research has demonstrated specific and practical changes physicians can make to improve their practices' effectiveness in immunizing children, including the following: 1) sending parent reminders for upcoming visits and recall notices; 2) using prompts during all office visits to remind parents and staff about immunizations needed at that visit; 3) repeatedly measuring practice-wide immunization rates over time as part of a quality improvement effort; and 4) having in place standing orders for registered nurses, physician assistants, and medical assistants to identify opportunities to administer vaccines . Pediatricians should work individually and collectively at local and national levels to ensure that all children receive all childhood immunizations on time . Pediatricians also can proactively communicate with parents to ensure they understand the overall safety and efficacy of vaccines.

Pediatrics, 2003 Oct, 112(4), 815 - 20
Risk of anaphylaxis after vaccination of children and adolescents; Bohlke K et al.; OBJECTIVE: To quantify the risk of anaphylaxis after vaccination of children and adolescents . METHODS: The study population consisted of children and adolescents who were enrolled at 4 health maintenance organizations that participated in the Vaccine Safety Datalink Project . For the period 1991-1997, we identified potential cases by searching for occurrences of International Classification of Diseases, Ninth Revision (ICD-9) code 995.0 (anaphylactic shock), E948.0 through E948.9 (adverse reaction from bacterial vaccines), and E949.0 through E949.9 (adverse reaction from other vaccines and biological substances) . At 1 study site, we also included a range of other allergy codes . We restricted to diagnoses on days 0 to 2 after vaccination (ICD-9 995.0) or day 0 (all other ICD-9 codes) . We then reviewed the medical record to confirm the diagnosis . RESULTS: We identified 5 cases of potentially vaccine-associated anaphylaxis after administration of 7 644 049 vaccine doses, for a risk of 0.65 cases/million doses (95% confidence interval: 0.21-1.53) . None of the episodes resulted in death . Vaccines that were administered before the anaphylactic episodes were generally given in combination and included measles-mumps-rubella, hepatitis B, diphtheria-tetanus, diphtheria-tetanus-pertussis, Haemophilus influenzae type b, and oral polio vaccine . One case of anaphylaxis followed measles-mumps-rubella vaccine alone . At the site at which we reviewed additional allergy codes, we identified 1 case after 653 990 vaccine doses, for a risk of 1.53 cases/million doses (95% confidence interval: 0.04-8.52) . CONCLUSIONS: Patients and health care providers can be reassured that vaccine-associated anaphylaxis is a rare event . Nevertheless, providers should be prepared to provide immediate medical treatment should it occur.

N Engl J Med, 2003 Oct 2, 349(14), 1341 - 8
A trial of a 9-valent pneumococcal conjugate vaccine in children with and those without HIV infection; Klugman KP et al.; BACKGROUND: Acute respiratory tract infections caused by Streptococcus pneumoniae are a leading cause of morbidity and mortality in young children . We evaluated the efficacy of a 9-valent pneumococcal conjugate vaccine in a randomized, double-blind study in Soweto, South Africa . METHODS: At 6, 10, and 14 weeks of age, 19,922 children received the 9-valent pneumococcal polysaccharide vaccine conjugated to a noncatalytic cross-reacting mutant of diphtheria toxin (CRM197), and 19,914 received placebo . All children received Haemophilus influenzae type b conjugate vaccine . Efficacy and safety were analyzed according to the intention-to-treat principle . RESULTS: Among children without human immunodeficiency virus (HIV) infection, the vaccine reduced the incidence of a first episode of invasive pneumococcal disease due to serotypes included in the vaccine by 83 percent (95 percent confidence interval, 39 to 97; 17 cases among controls and 3 among vaccine recipients) . Among HIV-infected children, the efficacy was 65 percent (95 percent confidence interval, 24 to 86; 26 and 9 cases, respectively) . Among children without HIV infection, the vaccine reduced the incidence of first episodes of radiologically confirmed alveolar consolidation by 20 percent (95 percent confidence interval, 2 to 35; 212 cases in the control group and 169 in the vaccinated group) in the intention-to-treat analysis and by 25 percent (95 percent confidence interval, 4 to 41; 158 and 119 cases, respectively) in the per-protocol analysis (i.e., among fully vaccinated children) . The incidence of invasive pneumococcal disease caused by penicillin-resistant strains was reduced by 67 percent (95 percent confidence interval, 19 to 88; 21 cases in the control group and 7 in the vaccinated group), and that caused by strains resistant to trimethoprim-sulfamethoxazole was reduced by 56 percent (95 percent confidence interval, 16 to 78; 32 and 14 cases, respectively) . CONCLUSIONS: Vaccination with a 9-valent pneumococcal conjugate vaccine reduced the incidence of radiologically confirmed pneumonia . The vaccine also reduced the incidence of vaccine-serotype and antibiotic-resistant invasive pneumococcal disease among children with and those without HIV infection .

Expert Opin Pharmacother, 2003 Oct, 4(10), 1839 - 46
Amoxicillin/clavulanate potassium extended release tablets: a new antimicrobial for the treatment of acute bacterial sinusitis and community-acquired pneumonia; Benninger MS; Community-acquired bacterial respiratory tract infections are among the most common health disorders requiring medical care and are associated with substantial morbidity, mortality, and direct and indirect costs . Recent increases in the prevalence of antimicrobial resistance have resulted in reduced susceptibility of the most common respiratory tract bacterial pathogens to a number of antimicrobials . Amoxicillin/clavulanate potassium extended release (ER) tablets (Augmentin XR, GlaxoSmithKline) is a new formulation of amoxicillin/clavulanate that retains activity against betalactamase-producing organisms whilst increasing the activity against Streptococcus pneumoniae through elevated and sustained plasma amoxicillin concentrations . The bilayer tablet provides immediate release of clavulanate and both immediate and sustained release of amoxicillin to maintain therapeutic concentrations of amoxicillin over longer periods of the dosing interval . In clinical trials of acute bacterial sinusitis (ABS) and community-acquired pneumonia (CAP), amoxicillin/clavulanate ER was shown to have excellent bacteriological and clinical success rates, even in patients infected with antimicrobial-resistant pathogens, and was found to be generally well tolerated . Amoxicillin/clavulanate ER is approved in the US for the treatment of patients with ABS or CAP caused by beta-lactamase-producing pathogens (ie, Haemophilus influenzae, Moraxella catarrhalis, Haemophilus parainfluenzae, Klebsiella pneumoniae, or methicillin-susceptible Staphylococcus aureus) and S . pneumoniae with reduced susceptibility to penicillin (penicillin minimum inhibitory concentration = 2.0 microg/ml).

J Antimicrob Chemother, 2003 Nov, 52(5), 869 - 72 Epub 2003 Sep 30.
Comparative activity of garenoxacin and other agents by susceptibility and time-kill testing against Staphylococcus aureus, Streptococcus pyogenes and respiratory pathogens; Noviello S et al.; OBJECTIVES: Garenoxacin is a novel des-F(6)quinolone that has shown excellent antimicrobial activity against a wide range of clinically important microorganisms . In this study, its activity was examined, in comparison with that of other antimicrobial agents, by susceptibility and time-kill testing against Staphylococcus aureus, Streptococcus pyogenes and respiratory pathogens . METHODS: Overall, 200 bacterial strains were tested . The antimicrobial activity of garenoxacin was compared with that of ciprofloxacin, levofloxacin, moxifloxacin, amoxicillin, co-amoxiclav, cefuroxime, cefotaxime, ceftriaxone, imipenem, erythromycin and clarithromycin . In addition, the bactericidal activity of garenoxacin, moxifloxacin, levofloxacin and ciprofloxacin was evaluated by time-kill analysis against four strains each of staphylococci {two methicillin-susceptible (MSSA) and two methicillin-resistant (MRSA)}, pneumococci (two penicillin-susceptible and two penicillin-resistant) and Streptococcus pyogenes (two erythromycin-susceptible and two erythromycin-resistant) . Antibiotics were tested at concentrations 1-8 x MIC . RESULTS: MIC90 values of garenoxacin for the MSSA and MRSA strains were 0.03 and 2 mg/L, respectively . Among all the quinolones tested, garenoxacin yielded the lowest MIC values against all pneumococci (MIC90 0.12 mg/L) irrespective of macrolide resistance; the rank order of activity was garenoxacin> moxifloxacin>levofloxacin>ciprofloxacin . Excellent activity was shown also against Haemophilus influenzae (MIC90 <or= 0.03 mg/L) and Moraxella catarrhalis (MIC90 <or= 0.03 mg/L) . Ninety percent of S . pyogenes were inhibited at garenoxacin concentrations equal to 0.25 mg/L, its activity not being influenced by macrolide susceptibility . Garenoxacin was rapidly bactericidal against staphylococci, producing a >or= 3 log10 decrease in viable counts (cfu/mL) within 3 h at 4 x MIC, whereas a moderate, slower killing rate was observed versus streptococci . CONCLUSIONS: This investigational des-F(6)quinolone represents a promising alternative for the treatment of respiratory tract infections.

Vet Microbiol, 2003 Oct 17, 96(2), 189 - 202
A search for virulence genes of Haemophilus parasuis using differential display RT-PCR; Hill CE et al.; Although Haemophilus parasuis is an important bacterial pathogen of swine, little is known about its pathogenesis or why some strains seem to be more virulent than others . Therefore, we used differential display reverse transcription-polymerase chain reaction (DDRT-PCR) to search for virulence-associated genes in a pathogenic serotype 5 strain, H . parasuis 1185 . Gene expression was evaluated following growth in conditions chosen to begin to approximate those found in the upper respiratory tract and those encountered by the organism during acute infection . Seven different differentially expressed gene fragments were identified in cells grown at 40 degrees C in both the presence and absence of swine serum . Based on the deduced amino acid sequences, the most strongly up-regulated genes were homologs of fadD (a fatty acyl-CoA synthetase), apaH (diadenosine tetraphosphatase), pstI (enzyme I of the phosphoenolpyruvate-protein phosphotransferase system), and cysK (cysteine synthetase) . Homologs of Std (Na(+)- and Cl(-)-dependent ion transporter), HSPG (a mammalian basement membrane-specific heparin sulphate core protein precursor) and PntB (pyridine nucleotide transhydrogenase) were also up-regulated, but to a much lower extent . Sequences homologous to all of the differentially expressed genes were detected in the reference strains of all 15 H . parasuis serotypes . This is the first report of a global search for virulence factors of H . parasuis.

Nat Struct Biol, 2003 Nov, 10(11), 942 - 7 Epub 2003 Sep 28.
Structural determinants of SecB recognition by SecA in bacterial protein translocation; Zhou J et al.; SecB is a bacterial chaperone involved in directing pre-protein to the translocation pathway by its specific interaction with the peripheral membrane ATPase SecA . The SecB-binding site on SecA is located at its C terminus and consists of a stretch of highly conserved residues . The crystal structure of SecB in complex with the C-terminal 27 amino acids of SecA from Haemophilus influenzae shows that the SecA peptide is structured as a CCCH zinc-binding motif . One SecB tetramer is bound by two SecA peptides, and the interface involves primarily salt bridges and hydrogen bonding interactions . The structure explains the importance of the zinc-binding motif and conserved residues at the C terminus of SecA in its high-affinity binding with SecB . It also suggests a model of SecB-SecA interaction and its implication for the mechanism of pre-protein transfer in bacterial protein translocation.

Kansenshogaku Zasshi, 2003 Aug, 77(8), 631 - 4
{Continuous occurrence of two cases of Haemophilus influenzae type b meningitis caused by different strains in a day nursery}; Hoshino T et al.; Two cases of Haemophilus influenzae type b (Hib) meningitis were occurred continuously in a day nursery . Both isolates obtained from cerebrospinal fluids of 2 patients were beta lactamase producing amoxicillin/clavulanic acid sensitive strains . But biotypes and restriction fragment length polymorphism patterns by SmaI digested pulse-field gel electrophoresis of two isolates were different, respectively . Although it was already reported simultaneous occurrence of two cases of Hib meningitis caused by same strain through the nursery contact, none were reported by different strains . It was considered that two colonized strains had caused meningitis in two patients continuously.

Kansenshogaku Zasshi, 2003 Aug, 77(8), 606 - 10
{Evaluation of rapid urinary antigen test kit for Streptococcus pneumoniae in children with pneumonia or meningitis}; Sakata H; We evaluated rapid urinary antigen test kit for Streptococcus pneumoniae (Binax NOW S . pneumoniae) in 85 inpatients between February 2002 and November 2002 . Diseases of patients were pneumonia in 82 and meningitis in 3 . The age range of the patients was from 4 months to 14 years . We studied urinary antigen assay and culture of nasopharyngeal swab in all patients . Three infants with meningitis were measured liquor by the kit . Two infants with meningitis due to S . pneumoniae showed positive reactions in urine and liquor, but result of 1 infant with meningitis due to Haemophilus influenzae was negative . Of 82 patients with pneumonia, S . pneumoniae was isolated from 52 patients and the urinary antigen test was positive in 39 . Thirty-eight patients were isolated S . pneumoniae in 39 positive patients and 14 children were isolated it in 44 negative patients . Sensitivity in this test kit was 73.1% and specificity was 96.8% . This test is useful for children as well as adults.

Med Care, 2003 Oct, 41(10), 1142 - 52
The Pediatric Economic Database Evaluation (PEDE) Project: establishing a database to study trends in pediatric economic evaluation; Ungar WJ et al.; BACKGROUND: While standard methods for conducting economic evaluations have evolved, little attention has been paid to the conduct of these studies in special populations such as children . OBJECTIVES: To build a database of pediatric economic evaluations and to examine trends in publication characteristics over a 20-year period . RESEARCH DESIGN: The database was created through a multisource search strategy, manual review, application of inclusion/exclusion criteria, data extraction, and reliability assessment . Descriptive statistics were used to summarize trends in publication volume, disease category, intervention type, and age group between 1980 and 1999 . RESULTS: From an initial cut of 5600 citations identified from 12 journal databases, 787 were included as full pediatric economic evaluations . Volume of publications increased 7-fold between 1980 to 1984 and 1995 to 1999 from 61 to 440 citations per 5-year period . Most studies were performed in children aged 1 to 12 years, and studies in infants displayed an increasing frequency . The most common disease category was infective/parasitic, comprising 24% of studies . Studies of congenital anomalies and complications of pregnancy were also prominent . Although health prevention studies were the most prevalent, health treatment studies demonstrated an equal frequency in 1995 to 1999 . Most studies consisted of malaria control and vaccination strategies for hepatitis B, Haemophilus influenzae type B, measles, and varicella . CONCLUSIONS: The number of pediatric economic evaluations is steadily increasing with most publications representing health prevention interventions . The Pediatric Economic Database Evaluation (PEDE) Project database will be valuable to health researchers working in methods research and conducting systematic reviews.

Appl Biochem Biotechnol, 2003 Aug, 110(2), 91 - 100
Characterization of polysaccharide production of haemophilus influenzae Type b and its relationship to bacterial cell growth; Takagi M et al.; Haemophilus influenzae type b (Hib) causes invasive infections in infants and young children . Vaccines consisting of Hib capsular polysaccharide (polymer of ribosylribitol phosphate {PRP}) conjugated to a protein are effective in the prevention of such infections . The production of capsular polysaccharide type b was studied in three cultivation conditions: single, glucose pulse, and repeated batch . Specific polysaccharide production (Yp/x) was calculated for all experiments, showing the following values: 67 (single-batch cultivation), 71 (glucose pulse), 75 (repeated-batch cultivation, first batch), and 87 mg of PRP/g of dry cell weight (DCW) (repeated-batch cultivation, second batch) . Biomass concentration reached approximately 1.8 g of DCW/L, while polysaccharide concentration was about approximately 132 mg/L in the three fermentation runs . Polysaccharide synthesis is associated with cell growth in all studied conditions as established by Kono's analysis and Luedeking-Piret's model.

Afr J Med Med Sci, 2001 Dec, 30(4), 295 - 303
Bacterial pathogens and outcome determinants of childhood pyogenic meningitis in Ilorin, Nigeria; Johnson WB et al.; Empirical antimicrobial therapy remainsjustifiable in childhood pyogenic meningitis (CPM), but the continuing efficacy in a particular setting requires periodic microbiological surveillance . It was this need that informed the present five-year retrospective study of consecutive admissions for CPM at the Emergency Paediatric Unit (EPU) of the University of Ilorin Teaching Hospital, Ilorin, Nigeria . Of the 71 cerebro-spinal fluid {CSF} analyses, 41 (57.6%) were Gram-smear positive (GSP) . Gram-positive cocci (GPC) were identified in 23 (56.1%) smears, while 14 (34.2.%) had Gram-negative bacilli (GNB) . Only three (7.3%) had Gram-negative diplococci (GND) . Despite corroborative biochemical findings, the remaining 30 (42.3%), including two with tuberculous meningitis proved smear-negative . GPC cases had a mean age of 4.49 +/- 5.3yrs, GNB, 3.06 +/- 4.8yrs and GND, 4.47 +/- 4.9yrs . CSF isolates were made in 28 (39.4%) cases . Streptococcus pneumoniae accounted for a predominant 22 (78.6%) (P = 0.00), Haemophilus influenzae for 2 (7.1%), and Neisseria meningitidis for only 1 (3.5%) case . Whereas Strept . pneumoniae and H . influenzae isolates were uniformly sensitive to each of sultamicillin, cefuroxime, ceftriaxone and ceftazidime, 7.7% of Strept . pneumoniae were resistant to crystalline penicillin, 6.7% to ampicillin, and 69.2% to chloramphenicol; one of the two H . influenzae isolates was chloramphenicol-resistant . Amongst the 30 (42.3%) fatal cases, the length of stay was significantly shorter in GNB-positive cases (P = 0.045) . Mortality was significantly higher amongst cases with purulent/turbid CSF at admission (P = 0.03), and in those with CSF protein of >150mg/dl (P = 0.02) and glucose <1mg/dl (P = 0.047) . The present aetiological preponderance of GPC and Strept . pneumoniae in our study population, the high case-fatality, and the emerging resistance profile suggest the need for exploring additional control options including vaccination . We emphasize the need for periodic auditing of local antimicrobial policies in CPM.

Commun Dis Intell, 2003, 27(3), 324 - 41
Progress towards eliminating Hib in Australia: an evaluation of Haemophilus influenzae type b prevention in Australia, 1 July 1993 to 30 June 2000; Horby P et al.; The status of Haemophilus influenzae (Hib) disease and its prevention by vaccination was reviewed for the period 1997 to 2000 . This forms the background to a change in national vaccine policy, from the use of two Hib vaccines to the use of PRP-OMP only throughout Australia from May 2000 . Notifications of Hib in the 7-year period between 1993 and 2000 declined by 87 per cent among children 0-4 years of age; adjustment for likely under-reporting increase this to a 95 per cent reduction . Among age groups not included in the immunisation program, there was also a substantial decline in notified cases . Overall, a minimum 430 cases and 13 deaths were prevented by Hib immunisation annually in Australia . Enhanced Hib surveillance recorded 532 cases over seven years, with 353 in unvaccinated persons, 74 fulfilling criteria for true vaccine failure and 75 partially immunised . Of unvaccinated cases, 60 and 182 were eligible for routine and catch-up immunisation respectively . Although the overall incidence for 0-4 years of age declined from 15 to 1.2 cases per 100,000 population, the proportion of cases under six months of age increased from 11 per cent to 23 per cent . Overall vaccine effectiveness, estimated using data from the last five years of the program, was 83 per cent (95% CI 71-91%), increasing to 90 per cent (95% CI 83-94%) when adjusted for under-reporting to the Australian Childhood Immunisation Register . Among Aboriginal and Torres Strait Islander people, the incidence of invasive Hib disease fell from 4.6 cases per 100,000 population to 0.7 cases per 100,000 population but the proportion of cases now occurring among Aboriginal or Torres Strait Islander people increased significantly, from 7 to 15 per cent . The Hib immunisation program in Australia has been highly successful . Nevertheless, experience in Australia and elsewhere indicates that continued careful monitoring of Hib disease, with high quality laboratory surveillance, remains important.

DNA Seq, 2003 Jun, 14(3), 169 - 81
Cloning and characterization of the Actinobacillus pleuropneumoniae fur gene and its role in regulation of ApxI and AfuABC expression; Hsu YM et al.; The ferric uptake regulation (fur) gene was cloned and characterized from Actinobacillus pleuropneumoniae and it exhibited 97% amino acid sequence identity to the Haemophilus ducrey fur gene . The flanking regions of the fur gene included an upstream putative flavodoxin (fldA) gene and a downstream possible transmembrane protein gene of unknown function . A single promoter was identified by 5' rapid amplification of cDNA ends (RACE), but there were no sequences homologous to an Escherichia coli Fur box in the 5' upstream sequence . The A . pleuropneumoniae fur clone complemented an E . coli fur deletion mutant . Transcriptional analysis of the divergent promoters of the A . pleuropneumoniae toxin I operon (apxICABD)--and the Actinobacillus ferric uptake operon (afuABC) showed that Fur and calcium together positively regulated the transcription of apxICABD while Fur was a repressor for afuABC . Hemolytic activity was significantly induced by iron and calcium and Fur appeared to act as an activator under high calcium conditions and as a repressor under low calcium conditions . A possible regulator-binding site was suggested by the properties of a point mutation in 33 bp upstream of the apxIC gene . This point mutation affected ApxI and Afu expression in response to iron, calcium, or Fur . These results provide further proof that calcium and the A . pleuropneumoniae Fur protein play a role in the expression of ApxI and Afu.

Curr Opin Investig Drugs, 2003 Aug, 4(8), 953 - 8
Vaccines for Moraxella catarrhalis and non-typeable Haemophilus influenzae; McMichael JC et al.; The development of vaccines against non-typeable Haemophilus influenzae and Moraxella catarrhalis represents a difficult challenge . Both bacteria are mucosal surface pathogens and protection may require a mucosal immune response . In addition, the surface antigens of non-typeable Haemophilus influenzae are hypervariable and animal models of infection with these bacteria may not be predictive of human efficacy . Vaccine development has focused on conserved surface exposed antigens, including integral outer membrane proteins, pili and other attachment factors, membrane-associated proteins, and lipooligosaccharide-protein conjugates . Several vaccine candidates are described that are antigenically conserved among strains, elicit biologically functional antibodies, and have efficacy in animal models.

Am J Prev Med, 2003 Oct, 25(3), 245 - 50
Impact of the Advisory Committee on Immunization Practices' 4-day grace period in a low-income community; Irigoyen M et al.; BACKGROUND: In 2002, the Advisory Committee on Immunization Practices (ACIP) recommended vaccine doses administered < or = 4 days before the minimum age or interval be counted as valid . The study objective was to assess the impact of the 4-day grace period on the need for revaccination and associated costs in a low-income community, compared to standard practice (i.e., repeating all doses that fall outside current ACIP guidelines) . METHODS: From 1999 to 2001, semi-annual immunization assessments of 8293 randomly selected children, aged 19-35 months, were conducted at a 16-practice network serving an underserved community in New York City . Outcome measures were rates of antigen-specific invalid doses and number of children needing revaccination, with and without the 4-day grace period . Revaccination costs were based on the Vaccines for Children (VFC) price list . RESULTS: The 4-day grace period reduced the number of children needing revaccination from 17.1% to 12.0%, a drop of 30% . The rates of invalid doses decreased from 1.9% to 1.3%, a drop of 33% . Invalid doses for hepatitis B (HepB)-2 decreased by two thirds (69.7%); for diphtheria-tetanus-acellular pertussis (DTaP)-1, Haemophilus influenzae type b (Hib)-1 and Hib-3 by half (44.9%-50.0%); for Polio-1, Polio-2, Polio-3, and measles-mumps-rubella (MMR) by one third (31.6%-33.3%); and for DTaP-2, DTaP-3, HepB-3, and varicella by nearly one quarter (20.0%-24.0%) . At these rates, revaccinating 100,000 children younger than age 3 years would cost 213,588 dollars per year, compared to 152,539 dollars with the 4-day grace period, in vaccine costs alone . CONCLUSIONS: In a low-income community, ACIP's 4-day grace period made a significant impact on the number of children requiring revaccination and on revaccination costs . However, the number of children needing revaccination remains high.

Antimicrob Agents Chemother, 2003 Oct, 47(10), 3270 - 4
Activities of a new oral streptogramin, XRP 2868, compared to those of other agents against Streptococcus pneumoniae and haemophilus species; Pankuch GA et al.; MIC methodology was used to test the antibacterial activity of XRP 2868, a new oral combination of two semisynthetic streptogramins, RPR 132552A and RPR 202868, compared to activities of other antibacterial agents against pneumococci, Haemophilus influenzae, and Haemophilus parainfluenzae . For 261 pneumococci, XRP 2868 and pristinamycin MICs were similar, irrespective of penicillin G and erythromycin A susceptibilities (MIC at which 50% of isolates were inhibited {MIC(50)}, 0.25 micro g/ml; MIC(90), 0.5 micro g/ml), while quinupristin/dalfopristin had MICs which were 1 to 2 dilutions higher . Single components of both XRP 2868 and quinupristin/dalfopristin had higher MICs . Erythromycin A, azithromycin, clarithromycin, and clindamycin MICs were higher for penicillin G-intermediate and -resistant than -susceptible pneumococci . Against 150 H . influenzae strains, all compounds tested had unimodal MIC distributions . XRP 2868 had an overall MIC(50) of 0.25 micro g/ml and an MIC(90) of 1.0 micro g/ml, with no differences between beta-lactamase-positive, beta-lactamase-negative, and beta-lactamase-negative ampicillin-resistant strains . Of note was the similarly low activity of one of its components, RPR 132552A . Pristinamycin and quinupristin/dalfopristin had MICs of 0.125 to 8.0 micro g/ml; quinupristin alone had MICs of 8.0 to >64.0 micro g/ml, and dalfopristin had MICs of 1.0 to >64.0 micro g/ml . Erythromycin A, azithromycin, and clarithromycin had modal MICs of 4.0, 1.0, and 8.0 micro g/ml, respectively . MICs of all compounds against H . parainfluenzae were 1 to 2 dilutions higher than against H . influenzae . XRP 2868 showed potent activity against pneumococci and Haemophilus strains irrespective of their susceptibility to other agents.

Antimicrob Agents Chemother, 2003 Oct, 47(10), 3179 - 86
High-dose azithromycin versus high-dose amoxicillin-clavulanate for treatment of children with recurrent or persistent acute otitis media; Arrieta A et al.; Infants and young children, especially those in day care, are at risk for recurrent or persistent acute otitis media (AOM) . There are no data on oral alternatives to high-dose amoxicillin-clavulanate for treating AOM in these high-risk patients . In this double-blind, double-dummy multicenter clinical trial, we compared a novel, high-dose azithromycin regimen with high-dose amoxicillin-clavulanate for treatment of children with recurrent or persistent AOM . Three hundred four children were randomized; 300 received either high-dose azithromycin (20 mg/kg of body weight once a day for 3 days) or high-dose amoxicillin-clavulanate (90 mg/kg divided twice a day for 10 days) . Tympanocentesis was performed at baseline; clinical response was assessed at day 12 to 16 and day 28 to 32 . Two-thirds of patients were aged < or =2 years . A history of recurrent, persistent, or recurrent plus persistent AOM was noted in 67, 18, and 14% of patients, respectively . Pathogens were isolated from 163 of 296 intent-to-treat patients (55%) . At day 12 to 16, clinical success rates for azithromycin and amoxicillin-clavulanate were comparable for all patients (86 versus 84%, respectively) and for children aged < or =2 years (85 versus 79%, respectively) . At day 28 to 32, clinical success rates for azithromycin were superior to those for amoxicillin-clavulanate for all patients (72 versus 61%, respectively; P = 0.047) and for those aged < or =2 years (68 versus 51%, respectively; P = 0.017) . Per-pathogen clinical efficacy against Streptococcus pneumoniae and Haemophilus influenzae was comparable between the two regimens . The rates of treatment-related adverse events for azithromycin and amoxicillin-clavulanate were 32 and 42%, respectively (P = 0.095) . Corresponding compliance rates were 99 and 93%, respectively (P = 0.018) . These data demonstrate the efficacy and safety of high-dose azithromycin for treating recurrent or persistent AOM.

Infect Immun, 2003 Oct, 71(10), 5994 - 6003
Inhibition of phagocytosis by Haemophilus ducreyi requires expression of the LspA1 and LspA2 proteins; Vakevainen M et al.; Haemophilus ducreyi previously has been shown to inhibit the phagocytosis of both secondary targets and itself by certain cells in vitro . Wild-type H . ducreyi strain 35000HP contains two genes, lspA1 and lspA2, whose encoded protein products are predicted to be 456 and 543 kDa, respectively . An isogenic mutant of H . ducreyi 35000HP with inactivated lspA1 and lspA2 genes has been shown to exhibit substantially decreased virulence in the temperature-dependent rabbit model for chancroid . This lspA1 lspA2 mutant was tested for its ability to inhibit phagocytosis of immunoglobulin G-opsonized particles by differentiated HL-60 and U-937 cells and by J774A.1 cells . The wild-type strain H . ducreyi 35000HP readily inhibited phagocytosis, whereas the lspA1 lspA2 mutant was unable to inhibit phagocytosis . Similarly, the wild-type strain was resistant to phagocytosis, whereas the lspA1 lspA2 mutant was readily engulfed by phagocytes . This inhibitory effect of wild-type H . ducreyi on phagocytic activity was primarily associated with live bacterial cells but could also be found, under certain conditions, in concentrated H . ducreyi culture supernatant fluids that lacked detectable outer membrane fragments . Both the wild-type strain and the lspA1 lspA2 mutant attached to phagocytes at similar levels . These results indicate that the LspA1 and LspA2 proteins of H . ducreyi are involved, directly or indirectly, in the antiphagocytic activity of this pathogen, and they provide a possible explanation for the greatly reduced virulence of the lspA1 lspA2 mutant.

Infect Immun, 2003 Oct, 71(10), 5523 - 30
Up-regulation of interleukin-8 by novel small cytoplasmic molecules of nontypeable Haemophilus influenzae via p38 and extracellular signal-regulated kinase pathways; Wang B et al.; Nontypeable Haemophilus influenzae (NTHI) is an important etiological agent of otitis media (OM) and of exacerbated chronic obstructive pulmonary diseases (COPD) . Inflammation is a hallmark of both diseases . Interleukin-8 (IL-8), one of the important inflammatory mediators, is induced by NTHI and may play a significant role in the pathogenesis of these diseases . Our studies demonstrated that a soluble cytoplasmic fraction (SCF) from NTHI induced much greater IL-8 expression by human epithelial cells than did NTHI lipooligosaccharides and envelope proteins . The IL-8-inducing activity was associated with molecules of < or =3 kDa from SCF and was peptidase and lipase sensitive, suggesting that small lipopeptides are responsible for the strong IL-8 induction . Moreover, multiple intracellular signaling pathways were activated in response to cytoplasmic molecules . The results indicated that the p38 mitogen-activated protein kinase (MAPK) and Src-dependent Raf-1-Mek1/2-extracellular signal-regulated kinase mitogen-activated protein kinase (ERK MAPK) pathways are required for NTHI-induced IL-8 production . In contrast, the phosphatidylinositol 3-kinase (PI3K)-Akt pathway did not affect IL-8 expression, although this pathway was concomitantly activated upon exposure to NTHI SCF . The PI3K-Akt pathway was also directly activated by IL-8 and significantly inhibited by an antagonist of IL-8 receptors during NTHI stimulation . These results indicated that the PI3K-Akt pathway is activated in response to IL-8 that is induced by NTHI and may lead to other important epithelial cell responses . This work provides insight into essential molecular and cellular events that may impact on the pathogenesis of OM and COPD and identifies rational targets for anti-inflammatory intervention.

Am J Vet Res, 2003 Sep, 64(9), 1176 - 80
Evaluation of survival of Actinobacillus pleuropneumoniae and Haemophilus parasuis in four liquid media and two swab specimen transport systems; del Rio ML et al.; OBJECTIVE: To determine duration and rates of recovery of Actinobacillus pleuropneumoniae and Haemophilus parasuis from 4 liquid media and 2 swab specimen transport systems and compare findings with those of Escherichia coli . SAMPLE POPULATION: One strain each of A pleuropneumoniae (biovar 1, serotype 1), H parasuis (serovar 5), and E coli (serotype O149:K91:H19) . PROCEDURE: Strains were incubated in brain heart infusion broth supplemented with horse serum and other nutrients or in horse serum alone, with and without nicotinamide-adenine dinucleotide in both instances, for 150 days at 4 degrees C or room temperature (21 degrees C) . Similarly, strains were tested in Stuart and Amies transport systems after storage at room temperature for 8 days . RESULTS: Colony counts greater than those of the initial inoculum were observed after incubation in horse serum for A pleuropneumoniae but not for H parasuis . Overall, incubation at 4 degrees C in the 4 liquid media resulted in longer recovery duration and higher rates than at room temperature . Culture of H parasuis resulted in lower recovery rates and shorter durations of recovery than culture of A pleuropneumoniae, except for culture in horse serum . Haemophilus parasuis survived longer than A pleuropneumoniae in the transport systems, and all organisms survived longer in the Amies system . CONCLUSIONS AND CLINICAL RELEVANCE: Survival of A pleuropneumoniae and H parasuis indicated that horse serum prolongs survivability, which may result in exposure of more animals during a prolonged period . The Amies system might be a good choice for collection of clinical samples from animals, especially for recovery of H parasuis.

Int J Syst Evol Microbiol, 2003 Sep, 53(Pt 5), 1449 - 56
Proposal of Histophilus somni gen . nov., sp . nov . for the three species incertae sedis 'Haemophilus somnus', 'Haemophilus agni' and 'Histophilus ovis'; Angen O et al.; Earlier investigations have shown that 'Haemophilus somnus', 'Haemophilus agni' and 'Histophilus ovis' represent the same species . In the present investigation, the taxonomic position of this species is explored further by sequencing the 16S rRNA and rpoB genes of strains that were investigated previously by DNA-DNA hybridization . These results clearly support the allocation of this species to a novel genus within the family PASTEURELLACEAE: The phenotypic separation of Histophilus somni gen . nov., sp . nov . from other members of the family can, for most strains, be based on capnophilia, yellowish pigmentation and indole production . However, due to phenotypic variation, the use of a species-specific PCR test based on the 16S rRNA gene is included in the species description . This is justified by the high sequence similarity of the 16S rRNA gene within the species and the fact that the highest sequence similarity to any other taxon within the family is 93.4 % . The type strain, 8025(T)=ATCC 43625(T)=CCUG 36157(T), was isolated in the USA from a bovine brain with lesions of thromboembolic meningoencephalitis.

Arch Bronconeumol, 2003 Sep, 39(9), 394 - 9
{Diagnostic value of quantitative cultures of endotracheal aspirate in ventilator-associated pneumonia: a multicenter study}; Valencia Arango M et al.; OBJECTIVE: To study the validity of quantitative cultures of tracheal aspirate (TA) in comparison with the plugged telescoping catheter (PTC) for the diagnosis of mechanical ventilator-associated pneumonia . METHOD: Prospective multicenter study enrolling patients undergoing mechanical ventilation for longer than 72 hours . TA samples were collected from patients with suspected ventilator-associated pneumonia, followed by PTC sampling . Quantitative cultures were performed on all samples . Patients were classified according to the presence or not of pneumonia, based on clinical and radiologic criteria, clinical course and autopsy findings . The cutoff points were > or = 103 colony-forming units (cfu)/mL for PTC cultures; the TA cutoffs analyzed were > or = 105 and > or = 106 cfu/mL . RESULTS: Of the 120 patients studied, 84 had diagnoses of pneumonia and 36 did not (controls) . The sensitivity values for TA > or = 106, TA > or = 105, and PTC, respectively, were 54% (95% confidence interval {CI}, 42%-64%), 71% (95% CI, 60%-81%), and 68% (95% CI, 57%-78%) . The specificity values were 75% (95% CI, 58%-88%), 58% (95% CI, 41%-74%), and 75% (95% CI, 58%-88%), respectively . Staphylococcus aureus was the microorganism most frequently isolated in both TA and PTC samples, followed in frequency by Pseudomomonas aeruginosa in TA samples and Haemophilus influenzae in PTC samples . No significant differences were found between the sensitivity of TA > or = 105 and that of PTC, nor between the specificities of TA > or = 106 and PTC . CONCLUSIONS: No differences in the specificities of PTC and TA were found when a TA cutoff of > or = 106 cfu/ml was used . Moreover, at a cutoff of > or = 105 the sensitivity of TA was not statistically different from that of PTC . Quantitative cultures of TA can be considered acceptable for the diagnosis of ventilator-associated pneumonia.

Arch Bronconeumol, 2003 Sep, 39(9), 387 - 93
{A comparison of moxifloxacin and amoxicillin in the treatment of community-acquired pneumonia in Latin America: results of a multicenter clinical trial}; Jardim JR et al.; Since community-acquired pneumonia (CAP) is a common disease with a high morbidity rate, it is important to obtain information concerning its etiology and susceptibility to antibiotics across different geographic areas . This study presents data obtained in 5 Latin American counties in the course of an international clinical trial that evaluated the efficacy and safety of treatment with either moxifloxacin or amoxicillin administered for 10 days to patients suspected of having CAP caused by a pneumococcal infection . Details are given of the pathogens identified, the patterns of sensitivity to antibiotics observed, and the clinical and microbiological results obtained.A total of 84 patients were studied, of whom 70 (83.3%) were evaluated at the end of the trial to determine the efficacy and safety of the treatment received . Gram-positive bacteria were found in samples from 29 patients (80.5%) . The pathogen was Streptococcus pneumoniae in 28 of those cases (77.7%) . Gram-negative bacteria were found in 7 patients (19.4%), the most common being Haemophilus influenzae in 3 patients (8.3%) . The presence of atypical microorganisms was detected in 18 of the 70 patients (25%), mainly Mycobacterium pneumoniae (n=11), and in 6 cases (8.5%) the infection was mixed . Ten strains of S . pneumoniae (35.7%) were shown to be susceptible to penicillin, 2 (7.1%) were highly resistant, and 16 (57.1%) showed moderate resistance . The clinical success rate at the final visit after treatment was 94.1% for moxifloxacin and 91.7% for amoxicillin . The results of this trial demonstrate a high prevalence of S . pneumoniae with reduced susceptibility to penicillin in patients with CAP in Latin America . It also revealed a high incidence of atypical pathogens and mixed infection in 8.6% of patients . This information should be taken into account when establishing protocols for empirical treatment of CAP in Latin America.

Rev Esp Quimioter, 2003 Jun, 16(2), 195 - 203
{Haemophilus influenzae and betalactam resistance: description of bla TEM gene deletion}; Molina JM et al.; The resistance to betalactam antibiotics in a total of 177 clinical isolates of Haemophilus influenzae, mostly from respiratory tract samples and characterized by their betalactamase production, was studied using the cephalosporin chromogenic assay and by detecting bla(TEM) and bla(ROB) genes . A substantial number of clinical isolates carrying the bla(TEM) gene, which presented a mutation consisting of the absence of a fragment of 136 base pairs, located upstream from the coding region including the 35 but not the 10 region of the promoter were found . This suggests that a new bla(TEM) promoter exists in these strains . This finding was associated with increased resistance to the antibiotics cefaclor and loracarbef compared with normal isolates . It was also found that 3.9% of the isolates carried the bla(ROB-1) gene.

Southeast Asian J Trop Med Public Health, 2003 Jun, 34(2), 249 - 53
Conjugate vaccines--a breakthrough in vaccine development; Makela PH; The encapsulated bacteria Streptococcus pneumoniae (the pneumococcus), Neisseria meningitidis (the meningococcus) and Haemophilus influenzae type b (Hib) are the main causes of purulent meningitis, the peak incidence of which is seen in the first two years of life . The polysaccharide capsule of these bacteria is an essential virulence determinant, and antibodies to it are protective, suggesting that a polysaccharide vaccine could prevent these diseases . The young child is, however, unable to respond with antibody production to these polysaccharides, making such vaccines useless in infancy . Conjugation of the polysaccharide to a protein carrier has proven a way to solve the problem . Immunization of infants with such a Hib conjugate vaccine was shown in 1987 to result in the desired antibody production and protection from Hib meningitis and bacteremia . The Hib vaccine is now a part of national infant immunization programs in large parts of Europe, the Americas and Australia, and has resulted in the virtual disappearance of Hib disease from these areas . A group C meningococcal and 7-valent pneumococcal vaccine, available since 2000, are likewise proving highly effective in preventing bacteremic disease . Further advantages of the conjugate vaccines are their ability to elicit immunologic memory and to reduce asymptomatic carriage of the bacteria, resulting in marked herd immunity . This paper was delivered as a lecture in January 2003 in Bangkok on the occasion of the Prince Mahidol Award for a life's work in the field of vaccinology.

Cell Microbiol, 2003 Oct, 5(10), 695 - 707
The Haemophilus ducreyi cytolethal distending toxin induces DNA double-strand breaks and promotes ATM-dependent activation of RhoA; Frisan T et al.; Among bacterial protein toxins, the cytolethal distending toxins (CDTs) are unique in their ability to activate the DNA damage checkpoint responses, causing cell cycle arrest or apoptosis in intoxicated cells . We provide direct evidence that natural intoxication of cells with the Haemophilus ducreyi CDT (HdCDT) holotoxin induces DNA double-strand breaks similarly to ionizing radiation . Upon DNA damage, epithelial cells and fibroblasts promote the formation of actin stress fibres via activation of the small GTPase RhoA . This phenomenon is not toxin specific, but is part of the ATM-induced cellular responses to genotoxic stresses, including ionizing radiation . Activation of RhoA is associated with prolonged cell survival, as HdCDT-treated epithelial cells expressing a dominant-negative form of RhoA detach and consequently die faster than cells expressing a functional RhoA . Our data highlight several novel aspects of CDT biology: (i) we show that a member of the CDT family causes DNA double-strand breaks in naturally intoxicated cells, acting as a true genotoxic agent; and (ii) we disclose the existence of a novel signalling pathway for intracellularly triggered activation of the RhoA GTPase via the ATM kinase in response to DNA damage, possibly required to prolong cell survival.

APMIS, 2003 Jun, 111(6), 653 - 7
Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus in systemic and nonoral infections in Finland; Paju S et al.; The oral cavity is the ecological niche for Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus, but occasionally they cause severe nonoral infections . In this study we present 20 systemic or nonoral infections due to A . actinomycetemcomitans and H . aphrophilus, comprising all isolates of these species forwarded to and stored in Finnish reference laboratories during the years 1988-2000 . The time from specimen collection to correct species identification was 9.3 days for A . actinomycetemcomitans and 10.7 days for H . aphrophilus . A . actinomycetemcomitans strains represented serotypes a, b, c, and d . Arbitrarily primed PCR distinguished four A . actinomycetemcomitans and six H . aphrophilus genotypes . Antimicrobial susceptibility testing with benzylpenicillin, amoxicillin, tetracycline, metronidazole, azithromycin, and trovafloxacin showed generally good activities against the present strains, and the susceptibility patterns closely resembled those of oral strains . The prolonged time to recover and identify A . actinomycetemcomitans and H . aphrophilus from systemic and nonoral infections may delay the correct diagnosis of the patient, but the good antimicrobial efficacies of antimicrobial agents against these pathogens give a good prognosis for the patients and advance the treatment of severe infections caused by these fastidious organisms of oral origin.

P N G Med J, 2002 Sep-Dec, 45(3-4), 213 - 8
Haemophilus influenzae type b meningitis: how much better is prevention than cure?
Duke T.
Haemophilus influenzae type b (Hib) is a major cause of meningitis and pneumonia in children . In Papua New Guinea (PNG) more than 20% of Hib are now resistant to chloramphenicol, and resistant Hib meningitis treated with chloramphenicol results in certain death or severe brain injury . Third-generation cephalosporins are a therapeutic option but are very expensive, while the Haemophilus influenzae type b conjugate vaccine would provide effective prevention . In a province of 380,000 people, using ceftriaxone as standard treatment for meningitis in all health facilities would only save an estimated 8 more lives per year than using chloramphenicol, and cost US dollars 1514 per additional life saved . Introduction of Hib vaccine would save, each year, 61 more lives than using chloramphenicol and 53 more lives than using ceftriaxone for meningitis treatment . The cost of a vaccination strategy for Hib meningitis would be US dollars 1216 for each of the 61 additional lives saved . Hib vaccine would be by far the most effective intervention to reduce mortality and severe neurological disability from Hib meningitis in PNG . Nationwide introduction of Hib vaccine is urgently needed, as antibiotics are now less effective in this disease than ever before.

J Vet Diagn Invest, 1999 Sep, 11(5), 416 - 22
Diagnosis of enzootic pneumonia in Danish cattle: reverse transcription-polymerase chain reaction assay for detection of bovine respiratory syncytial virus in naturally and experimentally infected cattle; Larsen LE et al.; A reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for detection of bovine respiratory syncytial virus (BRSV) in lung tissue of naturally and experimentally infected cattle . Primers were selected from the gene coding the F fusion protein, which is relatively conserved among BRSV isolates . The RT-PCR assay was highly specific, it yielded positive reactions only when performed on BRSV-infected cell cultures or tissues . The detection limit of the RT-PCR assay was assessed as 5 TCID50 . BRSV was detected in tissues of the respiratory tract and in the tracheobroncheal lymph node of calves euthanized 2-8 days after experimental infection with BRSV, whereas samples of other tissues and samples from mock-infected animals were negative at all time points . Examination of lung samples from 8 different regions of the lungs revealed that although the virus was most often found in the cranioventral lobules, it was frequently present in all lung lobules . Microbiologic examinations of all acute fatal cases of pneumonia (135 animals) in cattle submitted for diagnostic purposes during 1 year revealed that Actinomyces pyogenes (11%), Haemophilus somnus (10%), Pasteurella sp . (7%), and Pasteurella haemolytica (7%) were the most common bacterial agents found in the lungs . BRSV was identified using a conventional antigen enzyme-linked immunosorbent assay (ELISA) in 23 (17%) animals . The established BRSV-specific RT-PCR assay yielded positive results for the same 23 animals . In addition, 10 animals that were negative with the ELISA were positive with the RT-PCR assay . These results indicates that the RT-PCR assay can be a sensitive, reliable alternative to conventional diagnostic procedures.

Diagn Microbiol Infect Dis, 2003 Sep, 47(1), 373 - 6
Susceptibility trends of haemophilus influenzae and Moraxella catarrhalis against orally administered antimicrobial agents: five-year report from the SENTRY Antimicrobial Surveillance Program; Johnson DM et al.; The assessment of orally administered antimicrobial susceptibilities of common pathogens that cause community-acquired respiratory tract infections (CARTI) has become exceedingly important due to the number of office visits for this indication . Numerous local, regional and global studies have documented the susceptibilities of Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis, the most common CARTI pathogens . SENTRY Antimicrobial Surveillance Program sites in North and Latin America, and Europe were requested to send a combined total of 100 isolates of these pathogens to the local monitor for reference broth microdilution testing (1997-2001) . This study compared the susceptibility profiles of H . influenzae and M . catarrhalis isolates (13,370 strains) from the three geographic regions over a five year period . beta-lactamase mediated ampicillin resistance among H . influenzae was highest among North American isolates (27.9%) compared to Latin America and Europe (16.2 to 16.3%), although it was noted that during the five year study period, ampicillin resistance was steadily increasing in the latter two regions . Cefprozil (84.3% susceptible) and clarithromycin (81.1% susceptible) were also less active against North American H . influenzae isolates . Latin American isolates were much less susceptible to trimethoprim/sulfamethoxazole (T/S; 59.3%) compared to the other regions (75.8 to 78.6%) . M . catarrhalis isolates were also significantly less susceptible to T/S in Latin America (10.5% resistance) . The production of beta-lactamase enzymes among the M . catarrhalis isolates exceeded >95% in all three regions during the five year period . The fluoroquinolones (FQ) remained very active against these two respiratory pathogens with rare isolates with elevated FQ MIC results . It is apparent from this investigation that many commonly prescribed empiric treatments remain viable therapeutic options for CARTI caused by these two Gram-negative respiratory tract pathogens.

Proc R Soc Lond B Biol Sci, 2003 Aug 22, 270(1525), 1667 - 77
Epidemiology, hypermutation, within-host evolution and the virulence of Neisseria meningitidis; Meyers LA et al.; Many so-called pathogenic bacteria such as Neisseria meningitidis, Haemophilus influenzae, Staphylococcus aureus and Streptococcus pneumoniae are far more likely to colonize and maintain populations in healthy individuals asymptomatically than to cause disease . Disease is a dead-end for these bacteria: virulence shortens the window of time during which transmission to new hosts can occur and the subpopulations of bacteria actually responsible for disease, like those in the blood or cerebral spinal fluid, are rarely transmitted to new hosts . Hence, the virulence factors underlying their occasional pathogenicity must evolve in response to selection for something other than making their hosts sick . What are those selective pressures? We address this general question of the evolution of virulence in the context of phase shifting in N . meningitidis, a mutational process that turns specific genes on and off, and, in particular, contingency loci that code for virulence determinants such as pili, lipopolysaccharides, capsular polysaccharides and outer membrane proteins . We use mathematical models of the epidemiology and the within-host infection dynamics of N . meningitidis to make the case that rapid phase shifting evolves as an adaptation for colonization of diverse hosts and that the virulence of this bacterium is an inadvertent consequence of short-sighted within-host evolution, which is exasperated by the increased mutation rates associated with phase shifting . We present evidence for and suggest experimental and retrospective tests of these hypotheses.

J Int Med Res, 2003 Jul-Aug, 31(4), 307 - 11
Fine-needle aspiration as a diagnostic tool for recurrent tonsillitis; Inci E et al.; Acute tonsillitis is a common childhood disease, but repeated antibiotic treatment may fail, leading to tonsillectomy . Superficial swab cultures do not sufficiently represent the core bacteria present, but fine-needle aspiration may be a promising diagnostic method . We evaluated 58 patients undergoing tonsillectomy, and took superficial and core swabs, and fine-needle aspirations . Staphylococcus aureus was the most common bacterial pathogen identified, present in 26 out of 50 (52%) positive core cultures, followed by Haemophilus influenza in 13 positive core cultures (26%), and group A beta-haemolytic streptococci in 10 positive core cultures (20%) . Fine-needle aspiration detected 33 out of 50 positive core cultures (66%), whereas superficial swab culture detected 18 of 50 (36%); the difference being statistically significant . All pathogens detected by superficial swabs and fine-needle aspiration were detected in core cultures . The sensitivity and specificity of fine-needle aspiration were 66% and 100%, respectively, compared with 36% and 100% for superficial swabs . Fine-needle aspiration is therefore a promising method for detecting core bacteria in patients with recurrent tonsillitis.

Biochemistry, 2003 Sep 16, 42(36), 10756 - 63
Substrate specificity, metal binding properties, and spectroscopic characterization of the DapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase from Haemophilus influenzae; Bienvenue DL et al.; The catalytic and structural properties of divalent metal ion cofactor binding sites in the dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) from Haemophilus influenzae were investigated . Co(II)-substituted DapE enzyme was 25% more active than the Zn(II)-loaded form of the enzyme . Interestingly, Mn(II) can activate DapE, but only to approximately 20% of the Zn(II)-loaded enzyme . The order of the observed k(cat) values are Co(II) > Zn(II) > Cd(II) > Mn(II) >Ni(II) approximately equal Cu(II) approximately equal Mg(II) . DapE was shown to only hydrolyze L,L-N-succinyl-diaminopimelic acid (L,L-SDAP) and was inactive toward D,L-, L,D-, and D,D-SDAP . DapE was also inactive toward several acetylated amino acids as well as D,L-succinyl aminopimelate, which differs from the natural substrate, L,L-SDAP, by the absence of the amine group on the amino acid side chain . These data imply that the carboxylate of the succinyl moiety and the amine form important interactions with the active site of DapE . The affinity of DapE for one versus two Zn(II) ions differs by nearly 2.2 x 10(3) times (K(d1) = 0.14 microM vs K(d2) = 300 microM) . In addition, an Arrhenius plot was constructed from k(cat) values measured between 16 and 35 degrees C and was linear over this temperature range . The activation energy for {ZnZn(DapE)} was found to be 31 kJ/mol with the remaining thermodynamic parameters calculated at 25 degrees C being DeltaG(++) = 64 kJ/mol, DeltaH(++) = 28.5 kJ/mol, and DeltaS(++) = -119 J mol(-1) K(-1) . Electronic absorption and EPR spectra of {Co_(DapE)} and {CoCo(DapE)} indicate that the first Co(II) binding site is five-coordinate, while the second site is octahedral . In addition, any spin-spin interaction between the two Co(II) ions in {CoCo(DapE)} is very weak . The kinetic and spectroscopic data presented herein suggest that the DapE from H . influenzae has similar divalent metal binding properties to the aminopeptidase from Aeromonas proteolytica (AAP), and the observed divalent metal ion binding properties are discussed with respect to their catalytic roles in SDAP hydrolysis.

J Chemother, 2003 Aug, 15(4), 335 - 41
Comparative in vitro activity of telithromycin and beta-lactam antimicrobials against bacterial pathogens from community-acquired respiratory tract infections: data from the first year of PROTEKT (1999-2000); Kohno S et al.; The in vitro activity of telithromycin, a new ketolide, was compared with beta-lactam antimicrobials against pathogens commonly associated with community-acquired respiratory tract infections . These pathogens were collected during 1999-2000 as part of the ongoing PROTEKT surveillance study . Globally, penicillin nonsusceptibility among Streptococcus pneumoniae (n = 3362) was 36.3%, ranging from 21.5% (Australasia) to 68.0% (Far East) . Telithromycin showed higher potency (MIC90 0.12 mg/L) than beta-lactams against S . pneumoniae; 99.9% of all and 99.6% of multi-resistant isolates were susceptible to telithromycin . Among Streptococcus pyogenes isolates (n = 1485), 100% were susceptible to beta-lactams, and the telithromycin MIC50 and MIC90 were both 0.015 mg/L . Among Haemophilus influenzae (n = 2948), 16.6% produced beta-lactamase, which reduced the activity of ampicillin, cefaclor and cefprozil . 99.9% of H . influenzae were susceptible to telithromycin and the MIC range for M . catarrhalis was 0.004-0.5 mg/L . The first year results of PROTEKT confirmed high potency for telithromycin against common respiratory tract pathogens, including beta-lactam-resistant strains.

J Fish Dis, 2003 Jan, 26(1), 15 - 29
Analysis of exotoxins produced by atypical isolates of Aeromonas salmonicida, by enzymatic and serological methods; Gudmundsdottir BK et al.; In this study, exotoxins produced by 62 Aeromonas salmonicida strains and the bacterium Haemophilus piscium were analysed . Enzymatic assays, zymograms and serological detection were used to monitor secretion by bacterial strains of the previously described exotoxins P1, GCAT and AsaP1 and also the extracellular P2 metallo-gelatinase and a serine caseinase, which is different from the P1 protease and has not yet been characterized . Based on the results, the strains were divided into five groups . One comprised the type strains for A . salmonicida ssp . masoucida, H . piscium and 36% of the atypical isolates, and another, a type strain for A . salmonicida ssp . smithia together with 14% of the atypical isolates . A second type strain of A . salmonicida ssp . smithia was grouped with 8% of the atypical isolates . The largest group contained the type strains for A . salmonicida ssp . achromogenes and 38% of the atypical isolates . The type strains for A . salmonicida ssp . salmonicida were in the last group with all the four typical strains and 4% of the atypical isolates . The combination of zymogram and serological detection used is recommended as the most reliable method for characterizing A . salmonicida strains according to their exotoxin secretion.

J Clin Microbiol, 2003 Sep, 41(9), 4408 - 10
Chemically defined media for growth of Haemophilus influenzae strains; Coleman HN et al.; A chemically defined medium that supports the growth of both encapsulated and nontypeable Haemophilus influenzae strains in broth to densities that are >/= 10(9) CFU/ml or on agar plates is described . The mean generation time of a panel of clinical isolates was comparable to that in rich, chemically undefined media (brain-heart infusion broth supplemented with heme and beta-NAD).

Clin Infect Dis, 2003 Sep 15, 37(6), 841 - 5 Epub 2003 Aug 28.
Infections due to Haemophilus influenzae serotype E: microbiological, clinical, and epidemiological features; Campos J et al.; Surveillance after introduction of Haemophilus influenzae serotype b vaccination in Spain identified 26 H . influenzae serotype e (HiE) isolates . Of these, 16 (61.5%) were recovered from patients aged >16 years and 10 (38.5%) from children <16 years of age . HiE caused respiratory infections in 14 patients (9 with pneumonia), conjunctivitis in 4, vaginitis in 2, abscess in 2, and cellulitis, peritoneal infection, sepsis and meningitis in 1 patient each . HiE was strongly clonal and highly resistant to ampicillin and cotrimoxazole, and the incidence of HiE infection did not increase over time.

Acta Otolaryngol, 2003 Aug, 123(6), 724 - 9
Is hydrogen peroxide responsible for the inhibitory activity of alpha-haemolytic streptococci sampled from the nasopharynx?
Tano K, Grahn Hakansson E, Wallbrandt P, Ronnqvist D, Holm SE, Hellstrom S.
OBJECTIVE: The inhibitory effect of alpha-haemolytic Streptococci (AHS) in vitro on the three commonest otitis media pathogens, Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, was previously investigated . The aim of this study was to determine the mechanism of this inhibitory activity . MATERIAL AND METHODS/RESULTS: When fractions of AHS filtrate were assayed to determine their inhibitory activity after size-exclusion chromatography, the inhibitory activity was found in the fractions with a low molecular weight . The inhibitory effect was completely reversed when catalase was added to the cell-free filtrate of AHS . A quantitative method also revealed high production (approximately 3 mmol/l) of hydrogen peroxide in the AHS filtrate with the best inhibitory activity . Electron microscopy of bacteria exposed to AHS filtrate with an inhibitory effect showed changes similar to bacteria exposed to hydrogen peroxide . CONCLUSIONS: We conclude that the inhibitory effect of AHS is most likely due to the production of hydrogen peroxide . The significance of hydrogen peroxide production of AHS is discussed in relation to the non-specific and specific mucosal defence systems.

J Antimicrob Chemother, 2003 Oct, 52(4), 695 - 8 Epub 2003 Sep 01.
High levels of multiple antibiotic resistance among 938 Haemophilus influenzae type b meningitis isolates from Cuba (1990-2002); Tamargo I et al.; OBJECTIVES: A national surveillance study to determine antimicrobial susceptibility in Haemophilus influenzae type b isolated from cerebrospinal fluid was carried out in Cuba from 1990 to 2002 . METHODS: Susceptibility to ampicillin, co-amoxiclav, cefotaxime, ceftriaxone, co-trimoxazole, tetracycline, chloramphenicol and rifampicin was tested by the microdilution method according to the NCCLS guidelines . RESULTS: The 34 participating laboratories recovered 938 consecutive, non-identical isolates . All the isolates were retrieved from children aged <5 years . The mean number of isolates collected by year in the pre-vaccination era (1990-1998) was 93; after vaccination, 57 isolates were reported in 1999, 31 in 2000, four in 2001 and five in 2002 . Resistance to ampicillin, co-trimoxazole, tetracycline and chloramphenicol was 46.3% (all beta-lactamase-positive), 51.3%, 33.2% and 44.0%, respectively . Ampicillin-resistant beta-lactamase-negative strains were not detected . All strains were susceptible to co-amoxiclav, cefotaxime, ceftriaxone and rifampicin . Ampicillin resistance was strongly associated with resistance to tetracycline, co-trimoxazole and chloramphenicol (P<0.001) . Multidrug resistance was present in 43.8% of isolates . The most prevalent phenotype was resistance to ampicillin/chloramphenicol/tetracycline/co-trimoxazole, which was detected in 29.2% of strains overall . An increase in the prevalence of resistance to these antibiotics was observed from 1990 to 2000 in the range 40.7%-54.8% for ampicillin, 40.1%-51.6% for chloramphenicol, 45.4%-58.1% for co-trimoxazole and 23%-45.2% for tetracycline . CONCLUSIONS: In Cuba, the widespread vaccination against Haemophilus influenzae type b prevented a large number of meningitis cases in children caused by strains resistant to multiple antibiotics.

J Antimicrob Chemother, 2003 Oct, 52(4), 651 - 5 Epub 2003 Sep 01.
Factors influencing the anti-inflammatory effect of dexamethasone therapy in experimental pneumococcal meningitis; Lutsar I et al.; Dexamethasone (DXM) interferes with the production of tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) and can thereby diminish the secondary inflammatory response that follows initiation of antibacterial therapy . A beneficial effect on the outcome of Haemophilus meningitis in children has been proven, but until recently the effect of DXM therapy in pneumococcal meningitis was uncertain . The aim of the present study was to evaluate factors that might influence the modulatory effect of DXM on the antibiotic-induced inflammatory response in a rabbit model of pneumococcal meningitis . DXM (1 mg/kg) was given intravenously 30 min before or 1 h after administration of a pneumococcal cell wall extract, or the first dose of ampicillin . In meningitis induced by cell wall extract, DXM therapy prevented the increase in cerebrospinal fluid (CSF) leucocyte and lactate concentrations, but only if given 30 min before the cell wall extract . In meningitis caused by live organisms, initiation of ampicillin therapy resulted in an increase in CSF TNF-alpha and lactate concentrations only in animals with initial CSF bacterial concentrations > or =5.6 log10 cfu/mL . In those animals, DXM therapy prevented significant elevations in CSF TNF-alpha {median change -184 pg/mL, -114 pg/mL versus +683 pg/mL with DXM (30 min before or 1 h after ampicillin) versus controls (no DXM), respectively, P=0.02} and lactate concentrations {median change -10.6 mmol/L, -1.5 mmol/L versus +14.3 mmol/L with DXM (30 min before or 1 h after ampicillin) versus controls (no DXM), respectively, P=0.01} . These effects were independent of the timing of DXM administration . In this model of experimental pneumococcal meningitis, an antibiotic-induced secondary inflammatory response in the CSF was demonstrated only in animals with high initial CSF bacterial concentrations (> or =5.6 log10 cfu/mL) . These effects were modulated by DXM therapy whether it was given 30 min before or 1 h after the first dose of ampicillin.

Bioorg Med Chem Lett, 2003 Oct 6, 13(19), 3133 - 6
Synthesis and SAR evaluation of oxadiazolopyrazines as selective Haemophilus influenzae antibacterial agents; Beebe X et al.; The parallel synthesis and antibacterial activity of 5-hydroxy{1,2,5} oxadiazolo{3,4-b}pyrazines is reported . The compounds were synthesized by condensing diaminofurazan with alpha-keto acids to give a variety of aryl-substituted analogues . Halogenated phenyl groups at C-6 give rise to the greatest Haemophilus influenzae antibacterial activity.

Microbiology, 2003 Sep, 149(Pt 9), 2645 - 52
Effect of enzyme I of the bacterial phosphoenolpyruvate : sugar phosphotransferase system (PTS) on virulence in a murine model; Kok M et al.; The phosphoenolpyruvate : sugar phosphotransferase system (PTS) catalyses translocation with concomitant phosphorylation of sugars and hexitols and it regulates metabolism in response to the availability of carbohydrates . The PTS forms an interface between energy and signal transduction and its inhibition is likely to have pleiotropic effects . It is present in about one-third of bacteria with fully sequenced genomes, including many common pathogens, but does not occur in eukaryotes . Enzyme I (ptsI) is the first component of the divergent protein phosphorylation cascade . ptsI deletions were constructed in Salmonella typhimurium, Staphylococcus aureus and Haemophilus influenzae and virulence of the mutants was characterized in an intraperitoneal mouse model . The log(attenuation) values were 2.3, 1.4 and 0.9 for the Sal . typhimurium, Sta . aureus and H . influenzae ptsI mutants, respectively . The degree of attenuation is correlated with the complexity of the respective PTS, which comprises approximately 40 components in Sal . typhimurium, but only 5 in H . influenzae.

J Bacteriol, 2003 Sep, 185(18), 5555 - 62
Glutathione and catalase provide overlapping defenses for protection against respiration-generated hydrogen peroxide in Haemophilus influenzae; Vergauwen B et al.; Glutathione is an abundant and ubiquitous low-molecular-weight thiol that may play a role in many cellular processes, including protection against the deleterious effects of reactive oxygen species . We address here the role of glutathione in protection against hydrogen peroxide (H2O2) in Haemophilus influenzae and show that glutathione and catalase provide overlapping defense systems . H . influenzae is naturally glutathione deficient and imports glutathione from the growth medium . Mutant H . influenzae lacking catalase and cultured in glutathione-deficient minimal medium is completely devoid of H2O2 scavenging activity and, accordingly, substantial amounts of H2O2 accumulate in the growth medium . H . influenzae generates H2O2 at rates similar to those reported for Escherichia coli, but the toxicity of this harmful metabolite is averted by glutathione-based H2O2 removal, which appears to be the primary system for protection against H2O2 endogenously generated during aerobic respiration . When H2O2 concentrations exceed low micromolar levels, the hktE gene-encoded catalase becomes the predominant scavenger . The requirement for glutathione in protection against oxidative stress is analogous to that in higher and lower eukaryotes but is unlike the situation in other bacteria in which glutathione is dispensable for aerobic growth during both normal and oxidative stress conditions.

Diagn Microbiol Infect Dis, 2003 Aug, 46(4), 285 - 9
Comparison of Streptococcus pneumoniae and Haemophilus influenzae susceptibilities from community-acquired respiratory tract infections and hospitalized patients with pneumonia: five-year results for the SENTRY Antimicrobial Surveillance Program; Gordon KA et al.; The SENTRY Antimicrobial Surveillance Program has been monitoring the activity of commonly prescribed and novel antimicrobial agents on a global scale from 1997 to the present . Specific objectives have documented the key resistance rates among pathogens from both patients hospitalized with pneumonia and those diagnosed with community-acquired pneumonia . Hemophilus influenzae and Streptococcus pneumoniae are common pathogens in both of these patient populations and the susceptibility profiles for these two species were compared to distinguish potential differences that may be evident in North American surveillance (1997-2001) . A total of 6,515 isolates of S . pneumoniae and 6,726 H . influenzae strains were tested using reference broth microdilution methods at a monitoring center . Ampicillin resistance was approximately 25% among H . influenzae isolates and did not significantly differ between strains from community-acquired infections or hospitalized patients . beta-lactamase-negative ampicillin resistant strains and fluoroquinolone refractory strains were rare (0.3 and </= 0.2%, respectively) and the former only detected among hospitalized patients . Macrolide-resistant H . influenzae was most prevalent in hospitalized patients with pneumonia (24.4% for clarithromycin) . In contrast, H . influenzae isolates from community-acquired infections were less susceptible (78.6-81.7%) to trimethoprim/sulfamethoxazole, but very susceptible to fluoroquinolones (99.9-100.0%) . The community-acquired S . pneumoniae isolates were generally more resistant to penicillin (16.7%) and other beta-lactams compared to those from hospitalized patients (12.1%), and were also much more resistant to trimethoprim/sulfamethoxazole (25.0%) compared to inpatient isolates (6.7%) . In contrast, isolates with reduced fluoroquinolone susceptibility or resistance were more common among hospitalized patients (ciprofloxacin resistance at 4.0%) and increased over the five monitored years . It is evident from this longitudinal study that some antimicrobial agents are becoming less efficacious against common respiratory tract pathogens depending on the clinical setting and surveillance of resistance appears to be a prudent practice.

J Struct Funct Genomics, 2003, 4(1), 1 - 9
Crystal structure of the YajQ protein from Haemophilus influenzae reveals a tandem of RNP-like domains; Teplyakov A et al.; A hypothetical protein encoded by the gene YajQ of Haemophilus influenzae was selected, as part of a structural genomics project, for X-ray crystallographic structure determination and analysis to assist with the functional assignment . The protein is present in most bacteria, but not in archaea or eukaryotes . The amino acid sequence has no homology to that of other proteins . The YajQ protein was cloned, expressed, and the crystal structure determined at 2.1-A resolution by applying the multiwavelength anomalous dispersion method to a mercury derivative . The polypeptide chain is folded into two domains with identical folding topology . Each domain has a four-stranded antiparallel beta-sheet flanked on one side by two alpha-helices . This structural motif is a characteristic feature of many RNA-binding proteins . The tetrameric structure observed in the crystal suggests a possibility of binding two stretches of double-stranded nucleic acid.

Jpn J Antibiot, 2003 Jun, 56(3), 163 - 70
{Antimicrobial activities of macrolides against recent clinical isolates, and analysis of resistant mechanisms}; Okubo T et al.; We examined antibacterial activities of 4 kinds of macrolides, erythromycin (EM), clarithromycin (CAM), azithromycin (AZM) and rokitamycin (RKM), against 6 bacterial species of clinical strains isoleted in 2002 . Bacterial isolates used were each 50 strains of methicillin-susceptible Staphylococcus aureus (MSSA), Streptococcus pyogenes, Streptococcus agalactiae, Moraxella (Branhamella) catarrhalis, Haemophilus influenzae and 43 strains of Streptococcus pneumoniae . S . agalactiae were derived from gynecological samples, and other species were isolated from respiratory specimens . Antimicrobial activities against S . aureus, S . pyogenes, S . agalactiae, M . catarrhalis and H . influenzae of 14-membered macrolides, such as EM and CAM, were higher than those of 16-membered macrolide, RKM . By contrast, against S . pneumoniae, RKM was more effective than 14-membered macrolides . Six, three and four strains of S . aureus, S . pyogenes and S . agalactiae, respectively, were resistant to macrolides . Thirty-five among 43 pneumococcal isolates were resistant, and 15 of the 35 were highly-resistant, MIC of > 128 micrograms/ml, to any one of EM, CAM or AZM . Isolation frequency of resistant strains to RKM was lower than those to 14- and 15-membered macrolides: only one strain was highly-resistant and 12 were intermediately-resistant . No resistant strain was recognized in M . catarrhalis and H . influenzae . Further, we analyzed the resistant mechanisms, methylation or efflux, of macrolide resistant strains by the double-disk method . Methylation was major mechanism in S . aureus, and in S . pyogenes, all of the resistance was caused by methylation . In S . agalactiae and S . pneumoniae, methylation and efflux shared about half and half.

Eur J Clin Microbiol Infect Dis, 2003 Sep, 22(9), 530 - 4 Epub 2003 Aug 26.
Chronic bronchitis in immunocompromised patients: association with a novel Mycoplasma species; Webster D et al.; Patients with primary antibody deficiency are prone to recurrent bronchitis, often caused by nonencapsulated Haemophilus influenzae and streptococcal infection . Productive cough often persists even after elimination of these organisms with antibiotics . During an investigation into the cause of unexplained chronic bronchitis in these patients, a novel Mycoplasma species (designated A39) was first isolated from the sputum of a man with X-linked agammaglobulinaemia . Screening of sputa from a further 45 patients with primary antibody deficiency showed that 10 were positive for a similar organism using culture and/or a polymerase chain reaction based on sequences within the 16S ribosomal RNA gene . A comparison of the sequence data showed that the organism was distinct from but similar to Mycoplasma pneumoniae and other closely related mycoplasmas found in humans and animals . Electron microscopy showed some unique morphological characteristics . Although respiratory symptoms improved after elimination of A39 from the sputum of the patient with X-linked agammaglobulinaemia, further work is needed to establish the organism as a pathogen.

Eur J Clin Microbiol Infect Dis, 2003 Sep, 22(9), 519 - 24 Epub 2003 Aug 21.
Diagnostic accuracy of otitis media and tympanocentesis skills assessment among pediatricians; Pichichero ME; The comparative study presented here evaluated pediatricians from Italy, Greece, South Africa, and a reference group in the USA to determine (i) their ability to accurately diagnose acute otitis media (AOM) and otitis media with effusion (OME) using otoscopy, (ii) their knowledge of antibiotics, and (iii) their technical competence in performing tympanocentesis . The participants included 66 pediatricians from Italy, 115 from Greece, 36 from South Africa and 2,190 from the USA (reference group) . Each pediatrician viewed nine video-recorded otoscopic examinations of tympanic membranes, after which their ability to differentiate AOM, OME and normal was ascertained . Questions were posed regarding appropriate, pathogen-directed antibiotic selection for AOM . A mannequin model was used to assess the technical proficiency of each pediatrician in performing tympanocentesis . Results were recorded for each group as the mean percentage +/- standard deviation . The correct diagnosis was made by each group of pediatricians in the following frequencies: Italy, 54+/-27% (range, 18-94%); Greece, 36+/-12% (range, 23-56%); South Africa, 53+/-21% (range, 22-88%); and the USA, 51+/-11% (range, 29-72%) . The difference between results from Greece and the US reference group was statistically significant ( P=0.002) . Pediatricians from each group over-diagnosed AOM with the following frequencies: Italy, 18+/-19% (range, 2-49%); Greece, 34+/-13% (8-50%); South Africa, 23+/-14% (7-44%); and the US reference group, 26+/-19% (7-51%) . Pediatricians correctly selected an antibiotic recommended for treatment of AOM caused by drug-resistant Streptococcus pneumoniae as follows: Italy, 89%; Greece, 77%; South Africa, 82%; and the USA, 80% . For treatment of beta-lactamase-producing Haemophilus influenzae, the results were: Italy, 90%; Greece, 70%; South Africa, 81%; and the USA, 77% . Tympanocentesis was optimally performed by >/=86% of all pediatricians . The results indicate that pediatricians may often misdiagnose OME as AOM, but they select appropriate antibiotics about 80% of the time and can be trained to accurately perform tympanocentesis.

Antimicrob Agents Chemother, 2003 Sep, 47(9), 3008 - 11
In vitro activities of ABT-492, a new fluoroquinolone, against 155 aerobic and 171 anaerobic pathogens isolated from antral sinus puncture specimens from patients with sinusitis; Goldstein EJ et al.; ABT-492 exhibited excellent in vitro activities against all 326 aerobic and anaerobic antral puncture sinus isolates tested with MICs (in micrograms per milliliter) at which 90% of the isolates tested were inhibited as follows: Haemophilus influenzae, 0.001; Moraxella catarrhalis, 0.008; and Streptococcus pneumoniae, 0.015 . It was four- to sixfold more active than other fluoroquinolones, including against levofloxacin-resistant strains of S . pneumoniae, methicillin-resistant Staphylococcus aureus, and Prevotella species.

Chang Gung Med J, 2003 May, 26(5), 315 - 22
Safety and immunogenicity of a diphtheria, tetanus, and acellular pertussis-inactivated poliovirus vaccine/Haemophilus influenzae type B combination vaccine administered to Taiwanese infants at 2, 4, and 6 months of age; Lin TY et al.; BACKGROUND: Combined vaccines are urgently needed to ensure compliance with the increasing number of recommended vaccines for children . We evaluated the safety and antibody response to a diphtheria, tetanus, and acellular pertussis-inactivated poliovirus vaccine/Haemophilus influenzae type b (DTaP-IPV/Hib) combination vaccine administrated to infants at 2, 4, and 6 months of age . METHODS: Sixty healthy infants between 6 and 12 weeks of age were enrolled . One group of vaccines received the DTaP-IPV/Hib in a single injection, while another group concurrently received DTaP-IPV and Hib at separate injection sites . Solicited adverse events were monitored by parental observation and were recorded on a diary card . Levels of serum antibodies to DTaP and polyribosyl-ribitolphosphate-tetanus (PRP-T) antigens were collected before the first vaccine dose and 1 month after the third vaccine dose . RESULTS: The combined-injection group tended to have lower local reactions, and there was no increase in reactogenicity when compared with the separate-injection group . Seroconversion rates were 100% in both groups for all antigens, except for the anti-polio 2 antibody in the combined-injection group (96.4%) . The combined-injection group had lower antibody levels of PRP (8.45 microg/ml) than did the separate-injection group (20.61 microg/ml) . However, the percentage of vaccines achieving protective levels of antibody to PRP (> or = 0.15 microg/ml or > or = 1.0 microg/ml) was similar in both groups . CONCLUSIONS: DTaP-IPV/Hib may be safely and effectively administered to healthy infants, using a 2-, 4-, and 6-month vaccination schedule . This combined vaccine is cost-effective, more acceptable to parents and physicians, and minimizes distress to infants.

Infect Immun, 2003 Sep, 71(9), 5398 - 401
Nicotinamide ribosyl uptake mutants in Haemophilus influenzae; Herbert M et al.; The gene for the nicotinamide riboside (NR) transporter (pnuC) was identified in Haemophilus influenzae . A pnuC mutant had only residual NR uptake and could survive in vitro with high concentrations of NR, but could not survive in vivo . PnuC may represent a target for the development of inhibitors for preventing H . influenzae disease.

Indian J Med Res, 2003 Feb, 117, 81 - 7
Comparison of disc diffusion & E test methods with agar dilution for antimicrobial susceptibility testing of Haemophilus influenzae; Manoharan A et al.; BACKGROUND & OBJECTIVES: Reliable methods of detection of antimicrobial resistance are of paramount importance in the treatment and management of infections caused by Haemophilus influenzae . The objective of the present study was to compare and evaluate the performance of disc diffusion and E test (Epsilometer test) with agar dilution method for antimicrobial susceptibility testing of H . influenzae . METHODS: A total of 46 isolates of H . influenzae from various invasive sites were included as test strains . Antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion method for ampicillin, chloramphenicol, trimethoprim-sulphamethoxazole (TMP-SMZ) and cefotaxime . Minimum Inhibitory Concentration (MIC) determination was performed by E test and agar dilution for the same set of antimicrobials . All tests were performed on Haemophilus test medium (HTM) . RESULTS: Disc diffusion showed a very major (2%) and minor (4%) interpretative error with TMP-SMZ and minor interpretative errors to ampicillin (13%) and chloramphenicol (24%) when compared to agar dilution method . E test produced only minor interpretative errors to chloramphenicol (7%) and TMP-SMZ (2%) and no interpretative errors with ampicillin and cefotaxime as against agar dilution . E test showed good agreement with agar dilution for each of the antimicrobial tested . INTERPRETATION & CONCLUSION: Disc diffusion test may be used as a preliminary screen for susceptibility testing of H . influenzae . E test is simple, easy to perform and a reliable method for determination of resistance in H . influenzae . However its cost and limited availability in India may limit its use . The reference agar dilution method can be used reliably in routine susceptibility testing of H . influenzae.

Microb Pathog, 2003 Sep, 35(3), 133 - 7
Specificity of antibodies directed against the cytolethal distending toxin of Haemophilus ducreyi in patients with chancroid; Mbwana J et al.; Antibodies specific for the cytolethal-distending toxin of Haemophilus ducreyi (HdCDT) complex and for the CdtA, CdtB, and CdtC components were measured by ELISA in the sera of 50 patients with culture and/or PCR proven chancroid, 42 patients with periodontitis, 50 blood donors from Tanzania, 50 blood donors from Sweden . In addition, the biological activity e.g . neutralization capacity of the sera were tested . Our results demonstrate that majority of chancroid patients and healthy individuals had detectable levels of serum antibodies to HdCDT complex and to separate toxin components . However, high levels (> or =100 units) of antibodies to HdCDT complex were significantly more prevalent in the sera of patients with both chancroid and periodontitis than in the sera of the corresponding controls (P=0.001 and P=0.04, respectively) . In the sera of the 50 patients with chancroid, antibodies to CdtA, CdtB, and CdtC were detected in 50, 35, and 34 individuals, respectively . Antibodies to CdtC, being less frequently detected than the antibodies to other components, show a good correlation with the neutralizing capacity of sera . High levels of neutralizing antibodies (> or =160) were detected in only 22 and 2% of the patients with chancroid and periodontitis, respectively . The data suggest that the low levels of anti-HdCDT antibodies, which include neutralizing antibodies, may contribute to limited protection in chancroid and since anti-HdCDT antibodies, may be detected in healthy individuals and in patients with certain disease conditions (e.g . periodontitis), they may not be specific markers for chancroid infection.

Przegl Epidemiol, 2003, 57(1), 57 - 65
{Meningitis and encephalitis in Poland in 2001}; Rosinska M et al.; 2367 cases of meningitis and 571 of encephalitis were reported in Poland in 2001 . Among meningitis 1324 cases were classified as viral and 838 as bacterial . Among encephalitis cases, 117 were classified as bacterial and 383 as viral . Etiological factor was determined in 419 (44%) cases of bacterial meningitis/encephalitis . Among them Neisseria meningitidis was found in 106 cases, Haemophilus influenzae in 75 cases and Streptococcus pneumoniae in 99 cases . As in the past type B was the predominant type of N . meningitidis cultured from patients . 210 cases of tick borne encephalitis were reported in Poland in 2001, most of them from endemic areas of north-eastern part of the country.

Clin Microbiol Infect, 2003 Jul, 9(7), 645 - 52
Microbiological etiology in clinically diagnosed community-acquired pneumonia in primary care in Orebro, Sweden; Lagerstrom F et al.; OBJECTIVE: To study the etiology of clinically diagnosed community-acquired pneumonia (CAP) in antibiotically naive patients attending a primary care center and treated at their homes . METHODS: A three-year prospective study was carried out, and 177 patients presenting with clinical signs of CAP were included . All patients had chest X-rays after inclusion, and 82 (46%) showed infiltrates . Nasopharyngeal swab culture was performed on all patients, and 51% produced a representative sputum sample . Paired sera were obtained from 176 patients . RESULTS: Among the 82 patients with radiographically proven CAP, Streptococcus pneumoniae was detected in 26 patients (32%), Haemophilus influenzae in 23 (28%), Mycoplasma pneumoniae in 15 (18%), and Chlamydia pneumoniae in four (5%) . Serologic evidence of a viral infection was found in 13 patients (16%) . Among the 95 patients without infiltrates, S . pneumoniae was found in 21 (22%), H . influenzae in 14 (15%), M . pneumoniae in two (2%), and C . pneumoniae in five (5%) . Viral infection was detected in 19 (20%) of these 95 patients . CONCLUSION: In primary care in Sweden, the initial antibiotic treatment in any patient with pneumonia should be effective against S . pneumonia and H . influenzae . In addition, M . pneumoniae should be targeted during recurrent epidemics . C . pneumoniae, and especially Legionella, seem to be uncommon in primary care.

Clin Microbiol Infect, 2003 Jul, 9(7), 590 - 9
In vitro susceptibility of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis: a European multicenter study during 2000-2001; Jones ME et al.; OBJECTIVE: To assess the current (2001) activity of respiratory fluoroquinolones and comparator agents against respiratory pathogens isolated in European countries . METHODS: During 2000-2001, we prospectively collected 1995 isolates of Haemophilus influenzae, 1870 isolates of Streptococcus pneumoniae and 649 isolates of Moraxella catarrhalis from hospital laboratories in France, Germany, Greece, Italy, Spain and the UK . National Committee for Clinical Laboratory Standards (NCCLS)-approved broth microdilution antimicrobial susceptibility testing methods and interpretive criteria were used throughout . RESULTS: Of the S . pneumoniae isolates, 99.6% were susceptible to moxifloxacin, gatifloxacin and levofloxacin; the corresponding figure for H . influenzae was 100% . All M . catarrhalis isolates had moxifloxacin MICs </= 0.12 mg/L . For all three pathogens, fluoroquinolone susceptibility remained unchanged from the previous 1997-98 study . The incidence of penicillin non-susceptibility in the S . pneumoniae isolates tested remained similar to or higher than that recorded in previous studies: France, 165/291 (56.7%); Germany, 46/506 (9.1%); Greece, 20/55 (36.4%); Italy, 45/364 (12.4%); Spain, 146/268 (54.5%); and the UK, 26/386 (6.7%) . Significant levels of resistance to oral compounds (cefuroxime, cefaclor, cefdinir, clarithromycin, azithromycin, tetracycline, and trimethoprim-sulfamethoxazole) were detected among S . pneumoniae isolates . beta-Lactamase production among H . influenzae isolates ranged from 6.2% to 33.1% per country, and ampicillin, clarithromycin or trimethoprim-sulfamethoxazole resistance were the most common phenotypes detected . beta-Lactamase production among M . catarrhalis isolates ranged from 94.1% to 100% per country . CONCLUSIONS: With the exception of a few localized reports, resistance to moxifloxacin and other new fluoroquinolones in common respiratory pathogens is a rare occurrence, despite significant resistance to other compound classes . Surveillance will play a key role in tracking changes in fluoroquinolone susceptibility in European countries.

Vaccine, 2003 Sep 8, 21(25-26), 3863 - 6
A pilot study to quantify parental anxiety associated with enrollment of an infant or toddler in a phase III vaccine trial; Langley JM et al.; We sought to measure the anxiety felt by parents at the time of entry into a randomized controlled vaccine trial, and to determine if anxiety level was associated with parental demographic variables or past experience . The children were 2-month-old infants entering a randomized controlled clinical trial (RCT) of a diphtheria-tetanus toxoid-acellular pertussis vaccine adsorbed with Haemophilus influenzae B conjugate, or toddlers enrolling in a RCT of a Meningococcal C conjugate vaccine . Nurses interviewed parents to collect demographic data and parents self-administered the Spielberger Self-evaluation Questionnaire (State Anxiety STAI-Y-I) {Manual for the State-Trait Anxiety Inventory (Form Y) (Self-evaluation Questionnaire), Consulting Psychologists Press Inc., Palo Alto, 1983}, a validated instrument measuring the temporary condition of "state anxiety." A regression tree (CART) (S-Plus) was used to identify factors associated with higher anxiety scores . Parents of 97 children enrolled . Anxiety scores ranged from 22.75 (lower anxiety) to 36.43 (higher anxiety) . The regression tree identified a structured tree with six branches . The highest anxiety scores occurred in fathers with education less then grade 8, mothers with education less than high school, birth order of the child less then the third, previous serious illness in the family, or lack of experience with research . In a group of parents agreeing to enroll their infant or toddler in a vaccine study, certain attributes and experiences were associated with higher anxiety at the time of immunization in the context of a RCT . These factors should be considered by vaccine researchers in the recruitment process of clinical trials.

Vaccine, 2003 Sep 8, 21(25-26), 3730 - 3
Immunological priming of one dose of inactivated hepatitis A vaccine given during the first year of life in presence of maternal antibodies; Lagos R et al.; BACKGROUND: In hepatitis A virus (HAV)-seronegative infants, inactivated hepatitis A vaccines are highly immunogenic . On the contrary, in infants who are HAV-seropositive before vaccination, the interfering effect of passively-transferred maternal anti-HAV antibodies leads to lower post-primary immunization anti-HAV levels, as compared to those achieved by seronegative infants . One possible way to overcome this drawback is to delay hepatitis A vaccination later during the first year of life . The objective of the study was to document the immunogenicity of an inactivated hepatitis A vaccine in 6 months old HAV-seropositive infants, given as two dose regimen consisting of a single primary immunization at 6 months of age, followed by a booster dose 6 months later . METHODS: The immunogenicity of one hepatitis A vaccine (Avaxim pediatric, Aventis Pasteur) was documented in 108 6 months old, HAV-seropositive infants randomly assigned to receive one priming dose of hepatitis A vaccine either concomitantly with (Group 2) or 2 weeks after the third dose of routine diphteria-tetanus-whole cell pertussis reconstituting lyophilized tetanus conjugated Haemophilus influenzae type b (DTwcP//PRP approximately T) vaccine and oral poliomyelitis vaccine (OPV) (Group 1) . A booster dose was given 6 months later, concomitantly with MMR vaccine . RESULTS: The 91 infants who were HAV-seropositive (ELISA titer >20 mIU/ml) at the moment of primo vaccination remained seropositive 1 month later . Geometric mean titers (GMT) decreased from 292 and 278 mIU/ml 1 month after the first dose, to 77.6 and 76.0 mIU/ml 6 months after, in Groups 1 and 2, respectively . Post-booster titers increased markedly in both groups, with GMTs of 1731 and 1866 mIU/ml and geometric mean post/pre-immunization titer ratios of 22.3 and 24.6, respectively . CONCLUSIONS: These results suggest that immunological priming induced by a single dose of Avaxim pediatric administered to 6 or 6.5 months old, HAV-seropositive infants is present and should not preclude the use of this vaccine in such populations.

Vaccine, 2003 Sep 8, 21(25-26), 3601 - 7
Interference of outer membrane protein PalA with protective immunity against Actinobacillus pleuropneumoniae infections in vaccinated pigs; van den Bosch H et al.; The role of antibodies to the outer membrane protein PalA of Actinobacillus pleuropneumoniae in protective immunity was studied in pigs vaccinated with purified PalA alone and PalA in combination with toxoids of the RTX toxins ApxI and ApxII using an established challenge model with the virulent serotype 1 of A . pleuropneumoniae . Pigs that developed antibody titers against PalA after immunization were more significantly affected by challenge with A . pleuropneumoniae serotype 1 . Following challenge, pigs that were immunized with PalA showed more severe respiratory symptoms, had a higher mortality rate and died faster . They also displayed much more severe lung lesions after necropsy than animals not immunized with PalA . Pigs that were immunized with toxoids of the two cytotoxins ApxI and ApxII were protected against challenge with A . pleuropneumoniae . In contrast, the protective efficacy of the ApxI and ApxII vaccine was completely lost when it was supplemented with PalA . Hence, antibodies induced against the outer membrane protein PalA of A . pleuropneumoniae aggravated the consequences of infection and counteracted the protective effect of anti-ApxI and anti-ApxII antibodies . Due to the high similarity between protein analogues of PalA from various bacteria of the Pasteurellaceae family such as P6 of Haemophilus influenzae or 16kDa Omp of Pasteurella multocida, this deleterious effect of PalA in vaccination should be taken into consideration in the development of vaccines against infections with other Pasteurellaceae.

Vaccine, 2003 Sep 8, 21(25-26), 3593 - 600
Comparison of the reactogenicity and immunogenicity of a combined diphtheria, tetanus, acellular pertussis, hepatitis B, inactivated polio (DTPa-HBV-IPV) vaccine, mixed with the Haemophilus influenzae type b (Hib) conjugate vaccine and administered as a single injection, with the DTPa-IPV/Hib and hepatitis B vaccines administered in two simultaneous injections to infants at 2, 4 and 6 months of age; Aristegui J et al.; An open, randomised, multicentre trial was performed to compare the reactogenicity and safety profile of the administration of a hexavalent diphtheria-tetanus-acellular pertussis-hepatitis B-inactivated polio (DTPa-HBV-IPV) vaccine administered in one injection mixed with Haemophilus influenzae type b (Hib) conjugate vaccine (Group 1) with that of a pentavalent DTPa-IPV vaccine mixed with a Hib vaccine (DTPa-IPV/Hib), simultaneously administered with HBV (Group 2) in two injections in opposite thighs, as a primary vaccination course, to healthy infants at 2, 4 and 6 months of age . A total of 235 completed the study, 120 from Group 1 and 115 from Group 2 . Blood samples (pre-vaccination and 1 month after the third dose) were obtained from a subset of infants (Group 1: 40; Group 2: 31) to assess the immune response to vaccination . Local and general solicited symptoms were recorded by parents on diary cards . Seven hundred and five diary cards (Group 1: 360; Group 2: 345) were collected . The clinically relevant and most commonly reported local reaction was pain (infant cried when the limb was moved) in 2.5% (Group 1) and 1.2% (Group 2) of diary cards . Fever was more frequently reported in Group 1 (21% of diary cards) than in Group 2 (12% of diary cards) . However only 3 and 2% of doses in Groups 1 and 2, respectively, were responsible for a rectal temperature between 38.6 and 39.5 degrees C and only one case (Group 2) had > or =39.5 degrees C . Other clinically relevant general symptoms were rarely recorded: irritability (2-2.8%), loss of appetite (0.3-0.6%) and drowsiness (0.3-0.3%) . All subjects included in the immunogenicity analysis had seroprotective titres to diphtheria, tetanus, polio virus types 1 and 3, Hib . Almost all subjects were seroprotected for anti-polio type 2 and hepatitis B (with the exception of 1 subject in Group 1 for each antigen) . The vaccines response rates to pertussis antigens were over 97 and 90% in Groups 1 and 2, respectively . This study shows that, from a clinical perspective, the DTPa-HBV-IPV/Hib vaccine given in a single injection has a similar reactogenicity and safety profile to that of two licensed vaccines (DTPa-IPV/Hib, HBV) given in two simultaneous injections to infants at 2, 4 and 6 months of age . This is a valuable advantage, since in some countries, such as Spain and the UK, an additional injection (for the administration of meningococcal C conjugate vaccine) has been recently included in the infants' vaccination calendars.

Clin Immunol, 2003 Aug, 108(2), 119 - 27
Molecular ontogeny of the human antibody repertoire to the Haemophilus influenzae type B polysaccharide: expression of canonical variable regions and their variants in vaccinated infants; Lucas AH et al.; A structurally conserved antibody combining site, encoded by the IGH V3-23 and kappa A2 variable (V) region gene segments, predominates the adult immune response to the Haemophilus influenzae type b (Hib) capsular polysaccharide (PS) . This site has been elevated to canonical status based upon its relative molecular uniformity and prevalence in adults . To date, no studies have examined the primary structure of Hib PS-specific antibodies in young infants, who are the primary targets of Hib vaccination . In this study we show that canonical Hib PS-specific heavy (H) and light (L) chain V regions are present in 4-month-old infants following two vaccinations with Hib PS-protein conjugates . The infant V regions contain sequence polymorphisms that resemble those found in adult antibodies, as well as polymorphisms at position 95a of the A2 L chain not previously observed in adults . In vitro studies of Fab fragments and recombinant IgG2 antibodies using these V regions identify sequence polymorphisms that impact Hib PS binding affinity and bactericidal activity . These results demonstrate the establishment of canonical V regions in early ontogeny and provide a structural explanation of how canonical antibodies in the infant can vary in their affinity and protective activity against Hib.

Turk J Pediatr, 2003 Apr-Jun, 45(2), 158 - 60
Primary haemophilus influenzae pyomyositis in an infant: a case report; Secmeer G et al.; Promyositis is a term used to denote primary pyogenic infection of the skeletal muscle . Because striped muscle tissue is normally resistant to bacterial infection, pyomyositis is very rare . In tropical countries, pyomyositis accounts for about 4% of hospital surgical admission, but it is far less common in temperate climates . It is more common in adults and especially in men, but it can occur at any age . We would like to present an 8-month-old infant to make pediatricians aware of the possibility of pyomyositis in cases of a mass over muscle, or of children complaining of joint pain or muscle aches even in the infancy period.

East Afr Med J, 2003 Apr, 80(4), 213 - 7
Antimicrobial susceptibility in community-acquired bacterial pneumonia in adults; Kariuki S et al.; OBJECTIVES: To determine the antimicrobial susceptibility patterns of Streptococcus pneumoniae and Haemophilus influenzae, two bacterial pathogens commonly associated with community-acquired pneumonia . DESIGN: Cross-sectional study . SETTING: Bacterial isolates were obtained from adults suspected to have community-acquired pneumonia and who sought treatment at two city council clinics in Nairobi, Kenya . Susceptibility to antimicrobial agents was performed using a microdilution broth method, according to the criteria set by the National Committee for Clinical Laboratory Standards . RESULTS: A total of 277 S . pneumoniae and 58 H . influenzae were obtained from 536 adults examined in the period January 1998 to December 1999 . Of the 277 S . pneumoniae, only 56.7% were susceptible to penicillin and 7.6% of strains were resistant to two or more antimicrobial agents . Of the 58 H . influenzae strains, 91.4% were sensitive to ampicillin, with 6.8% resistant to two or more antimicrobial agents . 8.6% were beta-lactamase producers and accounted for the entire ampicillin-resistant population . CONCLUSION: The prevalence of resistance to penicillin and other commonly used antibiotics among pneumococci is high and the large number of multi-resistant strains among H . influenzae is a cause for concern . The prudent use of antibiotics in treatment of pneumonia and other infections should be advocated to minimise spread of resistance.

Cochrane Database Syst Rev . 2003;(3):CD004305.
Corticosteroids in acute bacterial meningitis; van de Beek D et al.; BACKGROUND: Acute bacterial meningitis remains a disease with high mortality rate, ranging from 10 to 30 percent, despite advances in critical care . It has been estimated that between 5 to 40 percent of all patients can suffer hearing loss . The use of corticosteroids as adjuvant therapy in the treatment of acute bacterial meningitis is controversial despite several controlled clinical trials and three meta-analyses . In particular there are few data on the use of corticosteroids in adult meningitis . OBJECTIVES: We conducted a systematic review examining the efficacy and safety of adjuvant corticosteroid therapy in children and adults with acute bacterial meningitis . SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library issue 1, 2003)); MEDLINE (1966 to April 2002); EMBASE (1974 to April 2002); and HEALTHLINE (1988 to April 2002) and Current Contents for trials published before the April 1st 2002, and reference lists of articles . We also contacted manufacturers and researchers in the field . SELECTION CRITERIA: Eligible studies were published or non-published randomised controlled trials on corticosteroids as adjuvant therapy in acute bacterial meningitis . Patients of any age and in any clinical condition, treated with antibacterial agents and randomised to corticosteroid therapy (or placebo) of any type, could be included . At least case fatality rate or hearing loss had to be recorded for inclusion . DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data . Study authors were contacted for additional information . Adverse effects information was collected from the trials . MAIN RESULTS: Eighteen studies involving 1853 people were included . Overall, adjuvant corticosteroids were associated with lower case fatality (relative risk (RR) 0.76, 95% confidence intervals (CI) 0.59 to 0.98) and lower rates of both severe hearing loss (RR 0.36, 95% CI 0.22 to 0.60) and long-term neurological sequelae (RR 0.66, 95% CI 0.44 to 0.99) . In children, corticosteroids reduced severe hearing loss in bacterial meningitis caused by Haemophilus influenzae (RR 0.31, 95% CI 0.15 to 0.62), as well as in meningitis caused by other bacteria than H . influenzae (RR 0.42, 95%CI 0.20 to 0.89) . In adults there was a reduction in case-fatality (RR 0.38, 95% CI 0.18 to 0.78), however there were few data . Adverse events were not increased significantly with the use of corticosteroids . REVIEWER'S CONCLUSIONS: Adjuvant corticosteroids are beneficial in the treatment of children with acute bacterial meningitis . The limited data available in adults shows a trend in favour of adjuvant corticosteroids but a definite recommendation must await more studies . ERRATUM: During the review process of this systematic review the results of the European Dexamethasone in Adulthood Bacterial Meningitis Trial were published . (De Gans 2002) In this prospective, randomised, double-blind, multicenter trial, which included 301 adults with bacterial meningitis, treatment with dexamethasone was associated with a reduction in mortality (relative risk of death, 0.48; 95 CI 0.24 to 0.96; p = 0.04) . Therefore, dexamethasone should be given to all adults with bacterial meningitis and should be initiated before or with the first dose of antibiotics.

Cochrane Database Syst Rev . 2003;(3):CD001958.
Haemophilus influenzae oral whole cell vaccination for preventing acute exacerbations of chronic bronchitis; Foxwell AR et al.; BACKGROUND: Acute bronchitis leading to ongoing exacerbations is a serious condition predisposed to by viruses or bacteria . It can be fatal . Antibiotic therapy has not been particularly useful in clearing bacteria such as nontypeable Haemophilus influenzae (NTHi) because they colonise the upper respiratory tract . An oral NTHi vaccine has been developed to protect against recurrent acute episodes in chronic bronchitis . OBJECTIVES: To assess the effects of an oral whole cell nontypeable Haemophilus influenzae (NTHi) vaccine in protecting against recurrent acute episodes in chronic bronchitis . SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (issue 1, 2003); MEDLINE (1966 to 2003); EMBASE (1990 - 2003); Extramed (1994 to 2003); ISI Current Contents (1993 to 2003); Carl Uncover (1988 to 2003) and contacted investigators of the studies . SELECTION CRITERIA: Randomised trials comparing the effects of an oral monobacterial NTHi vaccine on patients with recurrent acute exacerbations of chronic bronchitis were included when there was overt matching of the vaccine and placebo groups on clinical grounds . DATA COLLECTION AND ANALYSIS: Three reviewers extracted data and assessed trial quality independently from original records and publications for incidence and severity of bronchitis episodes and carriage rate of nontypeable Haemophilus influenzae measured in the upper respiratory tract every three months following vaccination . MAIN RESULTS: Six trials were included in the study with a total of 440 participants . Oral vaccination using a monobacterial whole cell killed nontypeable Haemophilus influenzae significantly reduced the incidence of bronchitic episodes at three months after vaccination (Poisson rate ratio 0.666; 95% confidence interval {CI} 0.500, 0.887; p = 0.005) and perhaps at six months after vaccination (Poisson rate ratio 0.831; 95% CI 0.669, 1.031; p = 0.093) . The effect had disappeared by nine months . The severity of exacerbations in the treatment group, as measured by requirement to prescribe antibiotics, was likewise reduced by 58% at three months (Peto odds ratio = 0.42; 95% CI 0.16, 1.13), and by 65% at six months (Peto odds ratio = 0.35; 95% CI 0.16, 0.75) . REVIEWER'S CONCLUSIONS: Vaccination, in the autumn, of patients with recurrent acute exacerbations of chronic bronchitis reduced the number and severity of exacerbations over the winter months . A large clinical trial to assess longer term prognosis is needed.

Pediatr Med Chir, 2003 Mar-Apr, 25(2), 131 - 4
Low rates of antimicrobial resistance in respiratory pathogens from a pediatric population in north-eastern Italy; Busetti M et al.; BACKGROUND: Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes are the main agents of bacterial upper respiratory tract infections in infants and children . In the past decades, the proportion of patients infected with drug-resistant organisms has increased; surveying at a regional level is important for understanding the size of the problem and elaborating therapeutic guidelines based on local epidemiology . METHODS: Since 1997, all isolates of Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes have been surveyed for antimicrobial resistance . RESULTS: In our zone, we found 3.5% penicillin-resistant and 18% macrolide-resistant Streptococcus pneumoniae, 15.8% ampicillin-resistant Haemophilus influenzae and 24.8% erythromycin-resistant Streptococcus pyogenes . CONCLUSIONS: Penicillin-resistant Streptococcus pneumoniae (PRSP) in our area does not represent a real concern; conversely, macrolide resistance in Streptococcus pneumoniae is relatively frequent . Resistance to ampicillin in Haemophilus influenzae in our area seems relatively low, due to beta-lactamase production . Even for Streptococcus pyogenes, macrolide resistance rates in our region are relatively low if compared with other countries and other italian regions . In conclusion, in our area penicillins are still the first choice drugs, at least in noncomplicated or not recurrent infections.

J Med Microbiol, 2003 Sep, 52(Pt 9), 829 - 33
Bacteriocin-like inhibitory substance (BLIS) production by the normal flora of the nasopharynx: potential to protect against otitis media?
Walls T, Power D, Tagg J.
The normal bacterial flora of the upper airways provides an important barrier to invading pathogens . This study investigated the production of bacteriocin-like inhibitory substances (BLIS) by streptococci isolated from the nasopharyngeal flora of children who either do or do not experience recurrent acute otitis media (AOM) . Twenty children with recurrent AOM and 15 controls were tested . Swabs from the nasopharynx were evaluated for streptococci having BLIS activity against two representative strains of each of the AOM pathogens Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae and Moraxella catarrhalis . Streptococci displaying strong BLIS activity were characterized further and tested for known streptococcal bacteriocin structural genes . Sixty-five per cent of children had nasopharyngeal streptococcal isolates that were inhibitory to strains of one or more of the AOM pathogens . Six children (17 %) had streptococci that demonstrated strong BLIS activity against strains of at least three of the pathogenic species . Three of these inhibitory isolates were Streptococcus salivarius, two were S . pneumoniae and one was S . pyogenes . The inhibitory S . salivarius and S . pyogenes were shown to have structural genes for known streptococcal bacteriocins . No statistically significant difference was found between the two groups of children with respect to the presence of inhibitory streptococci in their nasopharyngeal floras . The finding of S . salivarius with strong inhibitory activity against several AOM pathogens in the nasopharyngeal flora of children is unique . Although there is no clear evidence from the present study that these organisms protect against AOM, their low pathogenicity and strong in-vitro BLIS production capability indicate that they should be incorporated in future trials of bacteriotherapy for recurrent AOM.

Prescrire Int, 2003 Aug, 12(66), 148 - 50
Antibiotics in children with acute otitis media?
Feasibility study of the immunogenicity and safety of a novel DTPw/Hib (PRP-T) Brazilian combination compared to a licensed vaccine in healthy children at 2, 4, and 6 months of age.
Instituto Carlos Chagas, Rio de Janeiro, RJ, BrazilVaccination of infants with conjugated Haemophilus influenzae type b (Hib) vaccines has been proven to reduce Hib meningitis by 95% and pneumoniae by 20% . The routine use of Hib vaccine is facilitated by the introduction of combination vaccines into the EPI (Expanded Plan of Immunization) . The objective of this study was to compare the immunogenicity and reactogenicity of an extemporaneously mixed DTPw/Hib (diphtheria-tetanus-whole cell pertussis) combination, using the technology of two Brazilian manufacturers, against a licensed DTPw/Hib European combination in 108 infants vaccinated at 2, 4 and 6 months according to the local national schedule . The Brazilian combination was highly immunogenic with Hib seroprotection rates (anti-PRP > 0.15 mg /ml of 98% after 2 doses and 100% after 3) . Also for tetanus and pertussis the new Brazilian combination was as immunogenic as the European counterpart, except the diphtheria seroprotection rates and titers were lower . There was also no clinically relevant difference in reactogenicity . If these feasibility results are confirmed, the Brazilian DTPw/Hib combination should help to boost the uptake of Hib vaccination in Brazil.

J Clin Microbiol, 2003 Aug, 41(8), 3915 - 8
Molecular epidemiology of Haemophilus influenzae type b isolated from children with clinical cases of conjugate vaccine failures; Campos J et al.; We analyzed eight invasive Haemophilus influenzae strains isolated from individual children vaccinated appropriately for their ages . Five of the strains were type b and three were nontypeable strains . Children infected with nontypeable strains had a protective level of anti-H . influenzae type b antibodies, while only one of the children whose cases represented true vaccine failure did.

J Clin Microbiol, 2003 Aug, 41(8), 3473 - 80
Typing of nonencapsulated haemophilus strains by repetitive-element sequence-based PCR using intergenic dyad sequences; Bruant G et al.; Intergenic dyad sequences (IDS) are short repeated elements that have been described for several Haemophilus genomes and for only two other bacterial genera . We developed a repetitive-element sequence-based PCR using an IDS-specific primer as a typing method (IDS-PCR) for nonencapsulated Haemophilus strains and compared this technique with pulsed-field gel electrophoresis (PFGE) of DNA restricted with SmaI . IDS-PCR was rapid, easy to perform, and reproducible, with a high discriminatory capacity for nontypeable Haemophilus influenzae (NTHI) strains . The 69 NTHI strains tested generated 65 different banding patterns . Epidemiologically related strains gave similar or identical fingerprints, and all of the unrelated strains except two showed different patterns . These results were in agreement with those obtained by PFGE . For 20 genital strains usually identified as being biotype IV NTHI and belonging to a cryptic genospecies of Haemophilus with remarkable genetic homogeneity, four bands were significantly present and six bands were significantly absent from the fingerprints . The 20 strains were gathered in 11 closely related profiles, whereas PFGE provided no band when DNA was treated with SmaI . IDS-PCR improved the differentiation previously obtained within this species by ribotyping and multilocus enzyme electrophoresis . Our findings suggest that IDS-PCR is a rapid, reliable, and discriminatory method for typing NTHI strains and is currently the most efficient method for distinguishing strains within the cryptic genospecies of HAEMOPHILUS:

Am J Respir Crit Care Med, 2003 Dec 1, 168(11), 1304 - 7 Epub 2003 Aug 06.
Bronchoscopic microsampling method for measuring drug concentration in epithelial lining fluid; Yamazaki K et al.; Direct measurement of the concentration of antimicrobial agents in bronchial epithelial lining fluid (ELF) would allow for a more informed approach to appropriate dosing of antimicrobial agents for respiratory tract infections . In this study, we determined the time versus concentration profile in ELF after an oral administration of levofloxacin, using recently developed bronchoscopic microsampling probes . These probes could be repeatedly and safely inserted through the fiberoptic bronchoscope in normal healthy volunteers . The concentration of levofloxacin in ELF was 43.4% of the corresponding serum value at 1 hour, reached the same level at 2 hours, decreased in a similar manner as that in serum, and returned to undetectable levels at 24 hours . It exceeded minimal inhibitory concentrations of Staphylococcus aureus (0.25 microg/ml), Klebsiella species (0.5 microg/ml), and Haemophilus influenzae (0.06 microg/ml) after 6 hours . The experimental procedure was well tolerated, and no complications were observed . In conclusion, bronchoscopic microsampling is a feasible and promising method for measuring antimicrobial concentrations in the target sites of respiratory tracts directly and repeatedly.

Lett Appl Microbiol, 2003, 37(3), 190 - 5
Rapid detection of Haemophilus influenzae by hel gene polymerase chain reaction; Yadav MC et al.; AIMS: To evaluate the efficiency of hel gene polymerase chain reaction (PCR) to detect Haemophilus influenzae in various clinical/non-clinical samples . METHODS AND RESULTS: Seventy-four clinical samples (cerebrospinal fluid, blood, sputum, throat and nasal swabs) and throat swabs of 17 asymptomatic carriers were collected . Primers were used to amplify the hel gene of H . influenzae encoding P4 outer membrane protein directly from the processed samples . The samples were also examined by conventional culture methods and the results were compared with those of PCR . The culture methods showed positive results in 60 (65.9%) of 91 samples in contrast to 62 (68.12%) samples tested positive by PCR . None of the culture-positive samples were PCR-negative while two of the culture-negative samples were PCR-positive . The specificity of the products was confirmed by Southern hybridization and failure of various other organisms to amplify the hel gene product . The sensitivity of the PCR assay was found to be 50 pg of DNA . CONCLUSIONS: These findings suggest that the hel gene PCR is a rapid, sensitive and a specific new method for direct identification of H . influenzae . SIGNIFICANCE AND IMPACT OF THE STUDY: Thus, this PCR test can improve the detection rate of H . influenzae in suspected clinical samples as compared with that of conventional culture methods.

J Anesth, 2003, 17(2), 84 - 91
Reduction of potential respiratory pathogens by oral hygienic treatment in patients undergoing endotracheal anesthesia; Okuda M et al.; PURPOSE: This study was conducted to evaluate the usefulness of mechanical and chemical prophylactic oral cleansing treatments for reducing potential respiratory pathogens existing in the oral cavity . METHODS: Thirty-two patients scheduled to undergo oral and maxillofacial surgery that required endotracheal anesthesia were randomly allocated to one of the two groups, the oral cleansing group (n = 16) or the noncleansing group (n = 16) . Culture and polymerase chain reaction (PCR) methods were used to detect and enumerate pathogens . Oral cleansing was carried out with an electric toothbrush capable of automatically supplying and aspirating povidone-iodine solution before surgery, followed by rinsing twice a day after surgery . Cephazolin (3 g x day(-1)) was given to all patients for 5 days after surgery . RESULTS: The PCR detection rates of Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, and Porphyromonas gingivalis in gargle samples before treatment were 87.5%, 68.8%, 53.1%, and 40.6%, respectively . Oral cleansing reduced the detection rates and numbers of methicillin-sensitive Staphylococcus species, S . pneumoniae, and H . influenzae . In contrast, there was no significant reduction of methicillin-resistant Staphylococcus species, S . pneumoniae, H . influenzae, or P . aeruginosa in subjects who underwent systemic cephazolin administration without oral cleansing . CONCLUSION: The combination of mechanical and chemical oral cleansing resulted in a significant reduction of potential respiratory pathogens in the oral cavity.

Sex Transm Infect, 2003 Aug, 79(4), 294 - 7
Expression of the cytolethal distending toxin in a geographically diverse collection of Haemophilus ducreyi clinical isolates; Kulkarni K et al.; OBJECTIVE: To screen a collection of isolates of Haemophilus ducreyi for expression of the cytolethal distending toxin (CDT) . METHODS: 45 clinical isolates of H ducreyi were screened for cytotoxic activity by examining the effect of culture supernatants on Hela cells . Expression was confirmed using immunoblotting with CDT specific monoclonal antibodies and the presence of the cdt genes determined by amplification of the cdt genes in a multiplex polymerase chain assay . RESULTS: Of the 45 clinical isolates, six isolates from differing geographical origins did not demonstrate cytotoxic activity . Expression of CDT was also not detected in these six isolates using immunoblotting and the genes cdtA, cdtB, and cdtC were not amplified using PCR . The remaining isolates demonstrated cytotoxic activity, expressed the CDT proteins, and the presence of the cdt genes was confirmed . CONCLUSIONS: CDT is considered a virulence factor of H ducreyi but was found to be absent in 13% of isolates from different geographical origins.

J Immunol, 2003 Aug 15, 171(4), 1978 - 83
The fourth surface-exposed region of the outer membrane protein P5-homologous adhesin of nontypable Haemophilus influenzae is an immunodominant but nonprotective decoying epitope; Novotny LA et al.; Nontypable Haemophilus influenzae is a major cause of otitis media and other mucosal infections . After natural disease in children and experimental disease in chinchillas, we found a hierarchical pattern of immunodominance among the four surface-exposed regions of the P5-homologous adhesin, with the greatest response directed to region 4 . However, Ab to region 4 is not protective . When this natural but biased response was refocused to region 3 by immunization, augmented bacterial clearance and protection from ascending otitis media was observed . Collectively, the data indicate that region 4 contains a highly immunodominant but nonprotective decoying epitope, the presence of which dampens the immune response to a subdominant but protective epitope in region 3.

Expert Rev Vaccines, 2002 Oct, 1(3), 399 - 410
Evolution of conjugate vaccines; Makela PH et al.; Conjugate vaccines--bacterial polysaccharides conjugated to proteins to improve their immunizing properties--have been a success story since their introduction less than 20 years ago . The Haemophilus influenzae type b conjugate vaccine has nearly eliminated invasive Haemophilus influenzae type b disease in large parts of the world . The key reasons for its success are its ability to induce immunologic memory and reduce asymptomatic carriage, hence the spread of infection . The first pneumococcal conjugate vaccine was licensed in 2000 and is already in wide demand in the USA . The first meningococcal conjugate vaccine was introduced in a nationwide program in the UK in 1999 . Current discussion focuses on the efficacy of the conjugate vaccines for different end-points and the cost-effectiveness of their use globally.

Expert Rev Vaccines, 2002 Oct, 1(3), 277 - 83
Immunogenicity and reactogenicity of DTPw-HB/Hib vaccine administered to colombian infants after a birth dose of hepatitis B vaccine; Lopez P et al.; OBJECTIVE: To evaluate, in an open study, the immunogenicity, safety and reactogenicity of a birth dose of hepatitis B vaccine followed by a three-dose course of diphtheria-tetanus whole-cell pertussis-hepatitis B vaccine, extemporaneously mixed with Haemophilus influenzae b (Hib) vaccine . METHODS: At 2, 4 and 6 months of age, a single group of 120 Colombian infants were enrolled in this study to receive a regimen consisting of three doses of the combination vaccine following a dose of hepatitis B vaccine at birth . RESULTS: Seroprotection/vaccine response rates to all vaccine antigens was 98-100% 1 month after completion of the full vaccination course . The vaccine had an acceptable reactogenicity profile and the incidence of reported local and general symptoms decreased with the administration of subsequent vaccine doses . CONCLUSION: The mixed DTPw-HB/Hib vaccine was safe and well-tolerated, with high immunogenicity against all component antigens . Compared with previous studies, reactogenicity did not increase with the additional dose of hepatitis B vaccine given at birth . The DTPw-HB/Hib combination can be used to provide primary vaccination of infants who have already received a first dose of hepatitis B vaccine at birth.

New Microbiol, 2003 Jul, 26(3), 281 - 8
Prevalence and chemosusceptibility of Neisseria meningitidis and Haemophilus influenzae in a population of central Italy; Cresti S et al.; The purpose of this study was to determine the prevalence of nasopharyngeal carriers of Neisseria meningitidis and Haemophilus influenzae, and to evaluate their chemoresistance . N . meningitidis was more frequently isolated in adolescents (10-15 years) . All of meningococci were susceptible to ceftriaxone and rifampin, only one isolate showed reduced susceptibility to chloramphenicol and four strains showed reduced penicillin susceptibility . The results show that these drugs are still effective for prophylaxis and treatment in our area . All strains of H . influenzae were susceptible to penicillin, ceftriaxone, chloramphenicol, rifampin, azithromicin and gentamicin . 6 nontypeable strains were resistant to sulfamethoxazole-trimethoprim, 7 strains of type a and c-f, and 3 non-typeable strains showed reduced susceptibility to tetracycline . In contrast with the current trend in the world, in our area the susceptibility of H . influenzae to betalactams was 100%, therefore these antibiotics are still the drugs of choice for treatment of invasive diseases.

J Theor Biol, 2003 Sep 7, 224(1), 87 - 96
Systemic metabolic reactions are obtained by singular value decomposition of genome-scale stoichiometric matrices; Famili I et al.; Genome-scale metabolic networks can be reconstructed . The systemic biochemical properties of these networks can now be studied . Here, genome-scale reconstructed metabolic networks were analysed using singular value decomposition (SVD) . All the individual biochemical conversions contained in a reconstructed metabolic network are described by a stoichiometric matrix (S) . SVD of S led to the definition of the underlying modes that characterize the overall biochemical conversions that take place in a network and rank-ordered their importance . The modes were shown to correspond to systemic biochemical reactions and they could be used to identify the groups and clusters of individual biochemical reactions that drive them . Comparative analysis of the Escherichia coli, Haemophilus influenzae, and Helicobacter pylori genome-scale metabolic networks showed that the four dominant modes in all three networks correspond to: (1) the conversion of ATP to ADP, (2) redox metabolism of NADP, (3) proton-motive force, and (4) inorganic phosphate metabolism . The sets of individual metabolic reactions deriving these systemic conversions, however, differed among the three organisms . Thus, we can now define systemic metabolic reactions, or eigen-reactions, for the study of systems biology of metabolism and have a basis for comparing the overall properties of genome-specific metabolic networks.

Cesk Slov Oftalmol, 2003 Jun, 59(3), 176 - 83
{Microbial inflammations of the orbit in children}; Ondracek O et al.; The author evaluates in a retrospective study the incidence, diagnosis and causal agent and therapeutic results of microbial inflammations of the orbit . During the period from Jan . 1 1997 till Dec . 31 1999 66 children were hospitalized with the diagnosis of inflammation of the orbit . The mean age of the children was 6.2 years, the mean period of hospitalization 10.2 days . Chandler's classification of inflammations of the orbit was used . The diagnosis was established in collaboration by an ophthalmologist, otolyryngologist and roentgenologist . The main cause of inflammations of the orbit was inflammation of the paranasal sinuses . The most frequent microbial agents were streptococci, staphylococci and Haemophilus influenzae . All patients were treated during hospitalization by antibiotics administered by the i.v . route . Surgery was necessary in 8 patients--in 4 patients with an orbital abscess and in 4 patients with a subperiostal abscess not responding to conservative treatment . The operation was performed by an otolaryngologist using always the endonasal approach . The authors did not record in their group any permanent ophthalmological complications resulting from the inflammation of the orbit.

J Infect Dis, 2003 Aug 15, 188(4), 481 - 5 Epub 2003 Jul 31.
Estimating Haemophilus influenzae type b vaccine effectiveness in England and Wales by use of the screening method; Ramsay ME et al.; In October 1992, Haemophilus influenzae type b (Hib) conjugate vaccine was introduced to infants in the United Kingdom with a "catch-up" program for those aged <4 years . Initially, the rate of invasive Hib disease decreased dramatically but has been increasing since 1999 . To determine possible reasons for this increase, the effectiveness of Hib conjugate vaccine was estimated by use of the screening method . Between October 1993 and December 2001, a total of 443 cases of Hib infection occurred in children eligible for vaccination; 363 (82%) were fully vaccinated . Vaccine effectiveness was estimated to be 56.7% (95% confidence interval, 42.5-67.4) . Effectiveness was lower in children vaccinated during infancy, compared with those who were vaccinated during the catch-up campaign (P=.0033), declined with time since vaccination (P=.0008), and was lower in children born during 2000-2002, compared with other children scheduled for infant vaccination (P=.0041) . Use of a catch-up vaccination program enhanced the control of Hib infection in England and Wales . Since 1999, however, low effectiveness in infants, declining effectiveness with age, and the use of lower-efficacy vaccines have contributed to increased rates of Hib infection . The potential role of boosters needs to be considered.

Laryngoscope, 2003 Aug, 113(8), 1378 - 85
The role of nuclear factor-kappa B in interleukin-8 expression by human adenoidal fibroblasts; Takaki M et al.; OBJECTIVES/HYPOTHESIS: The production of cytokines by adenoids is known to be associated with inflammation of nasopharynx and the pathogenesis of otitis media with effusion . However, the role of adenoids in producing inflammatory cytokines such as interleukin-8 (IL-8) is not yet clear . In the present study, expression of IL-8 in adenoidal fibroblasts was investigated at the level of transcription factors . Further, the effects of clarithromycin, a 14-member ring macrolide, on IL-8 gene expression and nuclear factor-kappa B (NF-kappa B) activation in adenoidal fibroblasts were evaluated . STUDY DESIGN: In vitro study for the production of inflammatory cytokine from human adenoidal fibroblasts . METHODS: Adenoidal fibroblasts were incubated with nontypeable Haemophilus influenzae endotoxin or interleukin-1 beta . Then the expression of IL-8 and the influence of NF-kappa B inhibitor and clarithromycin were evaluated . Interleukin-8 protein production was assessed by ELISA, and IL-8 messenger RNA production was measured by Northern blot analysis and reverse transcriptase-polymerase chain reaction . Activation of NF-kappa B and inhibition of its activation were determined by electrophoretic mobility shift assay . RESULTS: The expression of both IL-8 protein and messenger RNA in adenoidal fibroblasts was enhanced by Haemophilus influenzae endotoxin and interleukin-1 beta and was positively correlated with increases in NF-kappa B activity . Treatment of cells with the NF-kappa B inhibitor N-tosyl-(L)-phenylalanine chloromethyl ketone, as well as with clarithromycin, reduced expression of IL-8 and NF-kappa B activity in a dose-dependent manner . CONCLUSIONS: Results suggest that adenoidal fibroblasts produce IL-8 in response to endotoxin through NF-kappa B activation . The inhibitory effects of clarithromycin on NF-kappa B activation and IL-8 production in adenoidal fibroblasts might explain, in part, the mechanism of this drug in improving otitis media with effusion.

J Bacteriol, 2003 Aug, 185(16), 4930 - 7
Haemophilus influenzae Rd lacks a stringently conserved fatty acid biosynthetic enzyme and thermal control of membrane lipid composition; Wang H et al.; The organization of the fatty acid synthetic genes of Haemophilus influenzae Rd is remarkably similar to that of the paradigm organism, Escherichia coli K-12, except that no homologue of the E . coli fabF gene is present . This finding is unexpected, since fabF is very widely distributed among bacteria and is thought to be the generic 3-ketoacyl-acyl carrier protein (ACP) synthase active on long-chain-length substrates . However, H . influenzae Rd contains a homologue of the E . coli fabB gene, which encodes a 3-ketoacyl-ACP synthase required for unsaturated fatty acid synthesis, and it seemed possible that the H . influenzae FabB homologue might have acquired the functions of FabF . E . coli mutants lacking fabF function are unable to regulate the compositions of membrane phospholipids in response to growth temperature . We report in vivo evidence that the enzyme encoded by the H . influenzae fabB gene has properties essentially identical to those of E . coli FabB and lacks FabF activity . Therefore, H . influenzae grows without FabF function . Moreover, as predicted from studies of the E . coli fabF mutants, H . influenzae is unable to change the fatty acid compositions of its membrane phospholipids with growth temperature . We also demonstrate that the fabB gene of Vibrio cholerae El Tor N16961 does not contain a frameshift mutation as was previously reported.

Arch Bronconeumol, 2003 Aug, 39(8), 333 - 40
{Community-acquired pneumonia requiring hospitalization in immunocompetent elderly patients: clinical features, prognostic factors and treatment}; Saldias Penafiel F et al.; BACKGROUND: Community-acquired pneumonia (CAP) is the leading cause of death from infectious disease among the elderly . This study was carried out to examine the clinical manifestations, etiology, prognostic factors and treatment of CAP in immunocompetent elderly patients requiring hospitalization . RESULTS: A total of 306 elderly (80 +/- 7 years) patients were evaluated: 54% were male, 89% had concurrent diseases (principally cardiovascular and neurological disease, chronic obstructive lung disease and diabetes), and 97% were treated with second-or third-generation cephalosporins . Mean hospital stay was 10 days, and mortality was approximately 10% in hospital and around 13% on follow up at 30 days . As compared to younger CAP patients, multiple comorbidity, altered mental status, hypoxemia, high serum urea nitrogen on hospital admission were more frequent in the elderly . Intermediate care and intensive care unit admissions were also more frequent in the elderly . Hospital length of stay as well as mortality in the hospital and at 30 days were higher in elderly patients . The pathogen was identified as part of routine care in around 25% of cases . The most frequent pathogens were Streptococcus pneumoniae (10.5%), enteric Gram negative bacilli (5.2%), Staphylococcus aureus (4.2%) and Haemophilus influenzae (3.9%) . In multivariate analysis the prognostic factors on admission associated with in-hospital mortality were advanced age (> 83 years), absence of cough, low blood pressure and hyperphosphatemia . CONCLUSION: CAP in elderly patients is a prevalent disease with specific clinical and epidemiological characteristics, clinical course and prognosis.

Can J Vet Res, 2003 Jul, 67(3), 229 - 31
Levels of nicotinamide adenine dinucleotide in extracellular body fluids of pigs may be growth-limiting for Actinobacillus pleuropneumoniae and Haemophilus parasuis; O'Reilly T et al.; During infection, nutrient deprivation can alter bacterial phenotype . This, in turn, may have implications for pathogenesis and prophylaxis . Actinobacillus pleuropneumoniae (biotype 1) and Haemophilus parasuis, respiratory tract pathogens of swine, are both V-factor-dependent . The concentrations of V factor in the extracellular fluids of pigs are unknown and may limit the growth of these bacteria in vivo . The aim of this study was to determine the concentrations of nicotinamide adenine dinucleotide (NAD) in select porcine body fluids and to compare the availability of NAD in vivo with the affinities of the organisms for this compound . Levels in plasma, tissue fluids (peritoneal, pleural, synovial, and cerebrospinal), and laryngeal, tracheal, and lung washings were determined with an enzymatic cycling assay . We concluded that, although the NAD supply in the respiratory tract is probably not growth-limiting, it may become limiting if the organisms are disseminated.

J Antimicrob Chemother, 2003 Sep, 52(3), 371 - 4 Epub 2003 Jul 29.
Comparative activity of telithromycin against typical community-acquired respiratory pathogens; Buxbaum A et al.; OBJECTIVES: Respiratory tract infections (RTIs) remain a significant cause of morbidity and mortality . Major bacterial pathogens in RTIs, such as Streptococcus pneumoniae, have exhibited increasing resistance to a variety of antibiotics during the past decades . Telithromycin, the first ketolide, was designed especially to overcome this resistance . The present study was conducted to assess the comparative activity of telithromycin against typical RTI pathogens in Austria . METHODS: A total of 1,015 bacterial isolates was tested, including S . pneumoniae, Streptococcus pyogenes, Staphylococcus aureus and Haemophilus influenzae . MICs of the following antimicrobials: penicillin G, ampicillin (for H . influenzae), azithromycin, clarithromycin, erythromycin A and telithromycin were determined using the NCCLS broth microdilution method . RESULTS: Telithromycin showed excellent activity against S . pneumoniae, with 99.8% of all isolates being susceptible . Penicillin remained active with an MIC50 and MIC90 of 0.007 mg/L . Nevertheless, a notable increase in penicillin intermediate-resistant and resistant isolates, from 4.9% in 1996 to the present rate of 10%, was observed . There was also a distinct rise in the resistance levels of S . pneumoniae against the macrolides . All tested isolates of S . pyogenes were susceptible to penicillin and telithromycin, and only low levels of resistance against telithromycin were found in S . aureus (2.2%, MIC90 of 0.5 mg/L) . No telithromycin-resistant isolate of H . influenzae could be detected . CONCLUSIONS: This study demonstrates the rising prevalence of resistance among S . pneumoniae not only to penicillin but also to other antimicrobials . It also shows the value of telithromycin as an attractive option for the empirical treatment of community-acquired RTIs in an era of widespread antibacterial resistance.

Pediatr Infect Dis J, 2003 Jul, 22(7), 657 - 8
Impact of the introduction of a combined Haemophilus B conjugate vaccine and hepatitis B recombinant vaccine on vaccine coverage rats in a large West Coast health maintenance organization; Davis RL et al.; We evaluated the effect of introduction of a combined hepatitis B-Haemophilus influenzae b vaccine on vaccine coverage rates in a large health maintenance organization and found a modest improvement in up-to-date immunization status.

Zh Mikrobiol Epidemiol Immunobiol, 2003 May-Jun, (3), 71 - 6
{Bacterial infectious process in patients with chronic obliterating bronchiolitis}; Faustova ME et al.; Chronic obliterating bronchiolitis in children is characterized by the presence of chronic multifactor inflammatory infectious process with Streptococcus pneumoniae and Haemophilus influenzae being the leading causative agents . The inflammatory process is induced and maintained by S . pneumoniae and H . influenzae or by the association of these two pathogens, but the severity of the clinical course of obliterating bronchiolitis is determined by H . influenzae, which is confirmed by its high detection rate at the phase of exacerbation in patients with pronounced bronchial obstruction.

Zh Mikrobiol Epidemiol Immunobiol, 2003 May-Jun, (3), 27 - 30
{Postvaccinal immunity and immunological aspects of Haemophilus influenzae carrier state in children of different age groups after the administration of "Act-HIB" vaccine}; Gorbunov SG et al.; The article deals with H . influenzae (different serotypes) carrier state and immune response before and after the administration of the vaccine "Act-HIB" to children of different age groups . Children aged up to 1 year and over 1 year have been found to differ in the dynamics of carrier state and in the concentration of antibodies of different classes to the antigens of this infective agent, which makes it necessary to carry out their early immunization with a view to ensure their protection from H . influenzae infection.

Euro Surveill, 2003 Jun, 8(6), 139 - 44
Vaccine coverage of pre-school age children in France in 2000; Antona D et al.; This article presents results of the main measures on vaccine coverage carried out in France in children up to six years of age . Vaccine coverage is very high for diphtheria, tetanus, pertussis, and poliomyelitis, and satisfactory for vaccination against Haemophilus influenzae b invasive infections . It will be necessary, however, to increase vaccine coverage against measles, mumps and rubella in infants and to ensure efficient catch up . Hepatitis B vaccine coverage is deficient in infants and could be improved when the vaccine is available in a combined form.

Antimicrob Agents Chemother, 2003 Aug, 47(8), 2663 - 5
Efficacy of single-dose azithromycin in treatment of acute otitis media in children after a baseline tympanocentesis; Dunne MW et al.; Children with acute otitis media underwent tympanocentesis and were given a single dose of 30 mg of azithromycin/kg of body weight . At day 28, the overall clinical cure rate was 206 of 242 (85%) . Clinical cure rates for patients infected with Streptococcus pneumoniae (67 of 76; 88%) and Haemophilus influenzae (28 of 44; 64%) were consistent with historical rates for the 5-day dosing regimen.

Antimicrob Agents Chemother, 2003 Aug, 47(8), 2551 - 7
Haemophilus influenzae bla(ROB-1) mutations in hypermutagenic deltaampC Escherichia coli conferring resistance to cefotaxime and beta-lactamase inhibitors and increased susceptibility to cefaclor; Galan JC et al.; The clinical use of cefaclor has been shown to enrich Haemophilus influenzae populations harboring cefaclor-hydrolyzing ROB-1 beta-lactamase . Such a selective process may lead to the increased use of extended-spectrum cephalosporins or beta-lactams plus beta-lactamase inhibitors and, eventually, resistance to these agents, which has not previously been observed in H . influenzae . In order to establish which bla(ROB-1) mutations, if any, could confer resistance to extended-spectrum cephalosporins and/or to beta-lactamase inhibitors, a plasmid harboring bla(ROB-1) was transformed into hypermutagenic strain Escherichia coli GB20 (DeltaampC mutS::Tn10), and this construct was used in place of H . influenzae bla(ROB-1) . Strain GB20 with the cloned gene was submitted to serial passages in tubes containing broth with increasing concentrations of selected beta-lactams (cefotaxime or amoxicillin-clavulanate) . Different mutations in the bla(ROB-1) gene were obtained during the passages in the presence of the different concentrations of the selective agents . Mutants resistant to extended-spectrum cephalosporins harbored either the Leu169-->Ser169 or the Arg164-->Trp164 substitution or the double amino acid change Arg164-->Trp164 and Ala237-->Thr237 . ROB-1 mutants that were resistant to beta-lactams plus beta-lactamase inhibitors and that harbored the Arg244-->Cys244 or the Ser130-->Gly130 replacement were also obtained . The cefaclor-hydrolyzing efficiencies of the ROB-1 variants were strongly decreased in all mutants, suggesting that if bla(ROB-1) mutants were selected by cefaclor, this drug would prevent the further evolution of this beta-lactamase toward molecular forms able to resist extended-spectrum cephalosporins or beta-lactamase inhibitors.

Antimicrob Agents Chemother, 2003 Aug, 47(8), 2471 - 80
In vitro and in vivo activities of novel 2-(thiazol-2-ylthio)-1beta-methylcarbapenems with potent activities against multiresistant gram-positive bacteria; Ueda Y et al.; SM-197436, SM-232721, and SM-232724 are new 1beta-methylcarbapenems with a unique 4-substituted thiazol-2-ylthio moiety at the C-2 side chain . In agar dilution susceptibility testing these novel carbapenems were active against methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus epidermidis (MRSE) with a MIC(90) of </=4 micro g/ml . Furthermore, SM-232724 showed strong bactericidal activity against MRSA, in contrast to linezolid, which was bacteriostatic up to four times the MIC . SM-232724 showed good therapeutic efficacy comparable to those of vancomycin and linezolid against systemic infections of MRSA in cyclophosphamide-treated mice . The MICs of SM-197436, SM-232721, and SM-232724 for streptococci, including penicillin-intermediate and penicillin-resistant Streptococcus pneumoniae strains, ranged from </=0.063 to 0.5 micro g/ml . These drugs were the most active beta-lactams tested against Enterococcus faecium, and the MIC(90) s for ampicillin-resistant E . faecium ranged between 8 and 16 micro g/ml, which were slightly higher than the value for linezolid . However, time-kill assays revealed the superior bactericidal activity of SM-232724 compared to those of quinupristin-dalfopristin and linezolid against an E . faecium strain with a 4-log reduction in CFU at four times the MIC after 24 h of exposure to antibiotics . In addition, SM-232724 significantly reduced the numbers of bacteria in a murine abscess model with the E . faecium strain: its efficacy was superior to that of linezolid, although the MICs (2 micro g/ml) of these two agents are the same . Among gram-negative bacteria, these three carbapenems were highly active against Haemophilus influenzae (including ampicillin-resistant strains), Moraxella catarrhalis, and Bacteroides fragilis, and showed antibacterial activity equivalent to that of imipenem for Escherichia coli, Klebsiella pneumoniae, and Proteus spp . Thus, these new carbapenems are promising candidates for agents to treat nosocomial bacterial infections by gram-positive and gram-negative bacteria, especially multiresistant gram-positive cocci, including MRSA and vancomycin-resistant enterococci.

Transfus Apheresis Sci, 2003 Aug, 29(1), 39 - 44
Acute Chest Syndrome: the role of erythro-exchange in patients with sickle cell disease in Sicily; Lombardo T et al.; Acute Chest Syndrome (ACS) describes a syndrome characterized by the presence of a new pulmonary infiltrate on a chest X-ray, fever, and respiratory symptoms and is the leading cause of death and hospitalization in sickle cell disease (SCD) . We studied 21 patients affected by SCD (13 HbSbeta+, 4 HbS beta(o), 4 HbSS, mean age 38.2 years) . Six out of the 21 patients developed one episode of ACS (two patients had positive blood cultures, for Mycoplasma pneumoniae and Haemophilus influenzae respectively) . The aim of our study was to evaluate the therapeutic efficacy of red cell-exchange during ACS . This procedure decreases HbS levels . The patients who underwent erythro-exchange showed a dramatic clinical and radiographic improvement with stabilized HbS levels between 20% and 30% . During follow up (14-32 months), none of the 6 patients developed viral complications related to transfusion therapy, alloimmunization or recurrence of ACS . In conclusion, in regard to the pre- and post-red cell-exchange clinical and laboratory data, we can say that red cell-exchange provides a dramatic resolution of the episode of ACS, minimizes the development of iron overload, and rapidly decreases HbS and hematocrit levels . In light of our results, we hypothesize that ACS episodes are secondary to pulmonary damage and to a gradual worsening related to age, and that there is some evidence that individuals affected by SCD in the third to fourth decade of life are more susceptible to ACS and/or other severe disease-related complications, needing repeated and strict clinical follow up.

Expert Opin Pharmacother, 2003 Aug, 4(8), 1227 - 40
The management of bacterial meningitis in children; Duke T et al.; Bacterial meningitis is still a major cause of death and disability in children worldwide . With the advent of conjugate vaccines against the three major pathogens, the burden of disease is increasingly concentrated in developing countries that cannot afford the vaccines . Antibiotic resistance is an increasing problem; in developed countries, high-level resistance to beta-lactams among Streptococcus pneumoniae necessitates the addition of vancomycin to third-generation cephalosporins . In many developing countries, the problems are more fundamental . Increasing resistance of S . pneumoniae to penicillin and chloramphenicol and of Haemophilus influenzae to chloramphenicol means that many children with bacterial meningitis receive ineffective treatments, as third-generation cephalosporins are often unavailable or unaffordable . Case fatality rates are as high as 50% and neurological sequelae occur in one-third of survivors . The use of corticosteroids in meningitis is controversial; the evidence that they protect against neurological complications of childhood meningitis (particularly severe hearing loss) is strongest when: meningitis is caused by H . influenzae type b; dexamethasone is given before the first dose of antibiotics; a bactericidal antibiotic such as a third-generation cephalosporin is used; and in the early stages of the infection . There are few controlled clinical trials on which to base recommendations about other adjuvant therapy for meningitis . Avoidance of secondary brain injury from hypoxia, hypotension, hypo-osmolarity and cerebral oedema, hypoglycaemia or convulsions is essential for a good outcome . The problem of bacterial meningitis will only be solved if protein-conjugate vaccines (or other effective vaccine strategies) against S . pneumonia, H . influenzae and epidemic strains of Neisseria meningitidis are available to all the world's children . Making third-generation cephalosporins affordable in the developing world is also a necessary intervention, but better antibiotics will not overcome the problems of poor access to hospitals and late presentation with established brain injury, and will inevitably bring further pressure for antimicrobial resistance.

Infect Immun, 2003 Aug, 71(8), 4691 - 9
Efficacy of the 26-kilodalton outer membrane protein and two P5 fimbrin-derived immunogens to induce clearance of nontypeable Haemophilus influenzae from the rat middle ear and lungs as well as from the chinchilla middle ear and nasopharynx; Kyd JM et al.; The rat middle ear and lung clearance model has been used to show that the nontypeable Haemophilus influenzae 26-kDa outer membrane protein OMP26 is highly efficacious as a mucosal immunogen, inducing significantly enhanced clearance in immunized rats upon direct challenge of these two anatomic sites . Similarly, the chinchilla model of middle ear and nasopharyngeal clearance has been used to show that two P5 fimbrin adhesin-derived immunogens, LB1 and lipoprotein D (LPD)-LB1(f)(2,1,3), are highly efficacious as parenteral immunogens . Both induced significantly augmented clearance of nontypeable H . influenzae upon challenge of these sites . Here, these three nontypeable H . influenzae immunogens in addition to six bovine serum albumin and keyhole limpet hemocyanin conjugates of the synthetic peptide LB1(f) were assayed for relative efficacy in the reciprocal rodent model system . OMP26 was assayed in the chinchilla host by a parenteral immunization route, with clearance of the middle ear and nasopharynx used as outcome measures . Both LB1 and LPD-LB1(f)(2,1,3) were assayed in the rat host with a mucosal immunization route and clearance of nontypeable H . influenzae from the lungs and middle ears as outcome measures . Both of the immunogens were found to induce a high-titered and specific immune responses in the heterologous host system . Moreover, each was found to be highly efficacious in the reciprocal host system, providing strong support for the continued development and inclusion of both OMP26 and P5 fimbrin-derived peptides as candidate vaccine antigens directed at otitis media caused by nontypeable H . influenzae.

Biomedica, 2003 Jun, 23(2), 208 - 12
{Accuracy of the slide agglutination method evaluated with PCR in typing Haemophilus influenzae isolates}; Hidalgo M et al.; In 1998, the Colombian government initiated an immunization program for children under one year of age with a Haemophilus influenzae capsular type b conjugate vaccine . After two years, the surveillance program of the Colombian Instituto Nacional de Salud Microbiology Group reported a 40% decrease in meningitis cases caused by H . influenzae . This effect was attributed to the vaccination . The surveillance program uses the standardized slide agglutination technique to serotype H . influenzae . The current study validated the accuracy of the slide agglutination method by means of the PCR technique . From children under five years of age, 146 isolates were obtained . These were collected between 1999 and 2002, and were characterized by biochemical tests and serotyped by the INS as part of the surveillance program . PCR confirmed 93% of the H . influenzae serotype b and 92% of the other serotypes . When the slide agglutination technique is conducted under a strict quality control program, it remains a sensitive and specific tool for serotyping H . influenzae.

Biomedica, 2003 Jun, 23(2), 194 - 201
{Surveillance of Haemophilus influenzae serotypes and antimicrobial resistance in Colombia, 1994-2002}; Ovalle MV et al.; Invasive disease caused by Haemophilus influenzae serotype b results in high rates of morbidity and mortality among children . In 1994, the Microbiology Group at the Instituto Nacional de Salud (Colombia) initiated a program to detect antimicrobial resistance in H . influenzae . Invasive isolates were collected by hospitals and public health laboratories as part of surveillance programs for acute respiratory infections and acute bacterial meningitis . To determine the evolution of serotypes and antimicrobial resistance patterns, invasive H . influenzae isolates collected from 1994 to 2002 were compared, and the impact of Hib conjugated vaccine in Colombia was reassessed . The analysis included 683 isolates, 379 (55.5%) were recovered from male patients, 370 (54.2%) from children under one year, 227 (33.2%) from children aged 1 to 5, 19 (2.8%) from children aged 6 to 14, and 38 (5.6%) from children over 14 years; 29 (4.2%) with no age data . Clinical classification recorded 493 (72.2%) of the samples were from patients with meningitis, 181 (26.5%) with pneumonia, and 9 (0.9%) with other diseases . Eighty five percent of isolates corresponded to H . influenzae serotype b, 12.9% were non capsular, and 2.0% corresponded to other serotypes (10 a, 1 d, 1 e and 2 f) . Of the total number of isolates, 12.0% produced beta lactamase, 13.9% were resistant to ampicillin, 12.7% to trimethoprim sulfamethoxazole (SXT), 5.4% to chloramphenicol, 1% to cefuroxime . All isolates were susceptible to ceftriaxone . During the 10-year period, resistance to SXT increased from 5% to 13% . A significant decrease in meningitis cases was detected among children under one-year old and in the 1 to 5 age group . Before introducing the vaccine, an annual average of 43 and 23 isolates for each of these groups were received . During 2002, 10 and 6 isolates, respectively, were received for each group . Surveillance of invasive H . influenzae isolates has allowed the evaluation of Hib vaccine impact, as well as the detection of an increase of non-capsular isolates, and changes in resistance patterns.

Clin Otolaryngol, 2003 Aug, 28(4), 335 - 40
Serum IgA and IgG functional antibodies and their subclasses to Streptococcus pneumoniae capsular antigen found in two aged-matched cohorts of children with and without otitis media with effusion; Drake-Lee AB et al.; The relationship between acute otitis media and otitis media with effusion (OME) is uncertain and the aetiology of OME is multifactorial . Otitis media with effusion may be an inflammatory condition; both bacteria and viral infections could play a part in this inflammation . The four bacteria Streptococcus pneumoniae, Haemophilus influenza, Staphylococcus aureus and Branhamella catarrhalis cause 60% of the infections whereas S . pneumoniae accounts for up to 35% . IgA provides the dominant surface response to polysaccharide and lipopolysaccharide antigens, of which IgA2 is the main subclass . Once the mucosa has been breached, most protection is provided by IgG . IgG2 acts mainly against bacterial capsular antigens . This study looked at two groups of 50 children with and without OME who were aged between 3 and 10 years . The aims were to determine if, firstly, the levels of the serum immunoglobulins were different in the two groups, secondly whether these children made the appropriate antibody response to the capsular antigen to S . pneumoniae (PCP), and finally if there was a delay in the maturity of the IgA response . The total IgG, IgA and all subclass levels were measured using radial immunodiffusion . Levels of functional IgA and IgG were measured using ELISAs (25 patients in each group) . The results were analysed with non-parametric tests . The immunoglobulin levels were within the normal levels for both groups . There were very good correlations between the IgG total anti-PCP and the IgG2 anti-PCP (R > 0.9, p = 0.001) . There was a good correlation between the levels of both IgG total and IgG2 anti-PCP against IgA total anti-PCP in both groups (R > 0.85, p > 0.01) . This confirms a normal antibody response between both groups of patients . The ages of the controls and patients (50 samples) were correlated with increasing titres of circulating functional antibodies (P = 0.001) . This is highly suggestive of a normal age-related response . In conclusion, the findings were contradictory to our original hypothesis that there is a subtle difference in surface protection between children with and without OME . We believe that a previous history of recurrent acute otitis media is unrelated to the development of OME after 3 years of age.

Otolaryngol Head Neck Surg, 2003 Jul, 129(1), 5 - 10
Effect of amoxicillin or clindamycin on the adenoids bacterial flora; Brook I et al.; OBJECTIVE: We sought to compare the effect on the adenoid bacterial flora of patients with recurrent otitis media of antimicrobial therapy with amoxicillin (Am) or clindamycin (C).Patients and methods Forty-five children scheduled for elective adenoidectomy participated in a prospective randomized study . They were divided into 3 groups of 15 each to receive either no therapy (control) or 10 days of therapy with Am or C . Core adenoid tissues was quantitatively cultured for aerobic and anaerobic bacteria . RESULTS: Polymicrobial aerobic-anaerobic flora were present in all instances . The predominant aerobes in all groups were alpha-hemolytic and gamma-hemolytic streptococci, Haemophilus influenzae, Staphylococcus aureus, group A beta-hemolytic streptococci, and Moraxella catarrhalis . The prominent anaerobes were Peptostreptococcus, Prevotella, and Fusobacterium spp . The number of isolates was significantly reduced in those treated with Am (n = 110, P < 0.05) or C (n = 58, P < 0.001) compared with control (n = 148) . The number of bacteria per gram/tissue was lower in those treated with either antibiotics . The number of potential pathogens was lower in those treated with C compared with the other 2 groups (P < 0.001) . The number of beta-lactamase-producing bacteria was lower in those treated with C than in those treated with Am (P < 0.025) or control (P < 0.001) . CONCLUSIONS: These data illustrate the ability of C and, to a lesser degree, of Am to reduce the bacterial load as well as potential pathogens and beta-lactamase-producing bacteria from the adenoids of children with recurrent otitis media.

Eur J Biochem, 2003 Aug, 270(15), 3153 - 67
Structural analysis of lipopolysaccharides from Haemophilus influenzae serotype f . Structural diversity observed in three strains; Yildirim HH et al.; Structural elucidation of the lipopolysaccharide (LPS) from three serotype f Haemophilus influenzae clinical isolates RM6255, RM7290 and RM6252 has been achieved using NMR spectroscopy techniques and ESI-MS on O-deacylated LPS and core oligosaccharide material (OS) as well as ESI-MSn on permethylated dephosphorylated OS . This is the first study to report structural details on LPS from serotype f strains . We found that the LPSs of all strains were highly heterogeneous mixtures of glycoforms expressing the common H . influenzae structural element 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-Kdo-(2-->6)-lipid A with variable length of OS chains linked to each of the heptoses . The terminal heptose (HepIII) in RM6255 is substituted at the O-3 position by a beta-d-Glcp residue whereas HepIII in strains RM7290 and RM6252 is substituted at O-2 by the globoside unit (alpha-d-Galp-(1-->4)-beta-d-Galp-(1-->4)-beta-d-Glc) or truncated versions thereof . The central heptose (HepII) is substituted by an alpha-d-Galp-(1-->4)-beta-d-Galp-(1-->4)-beta-d-Glcp-(1-->4)-alpha-d-Glcp unit in RM7290 and RM6252 or truncated versions thereof . Strain RM6255 does not express galactose in its LPS and only shows a cellobiose unit elongating from HepII (beta-d-Glcp-(1-->4)-alpha-d-Glcp) . ESI-MSn on dephosphorylated and permethylated OS provided information on the existence of additional minor isomeric glycoforms.

Scand J Immunol, 2003 Aug, 58(2), 119 - 28
Mucosal vaccination against encapsulated respiratory bacteria--new potentials for conjugate vaccines?
Jakobsen H, Jonsdottir I.
Polysaccharide (PS)-encapsulated bacteria such as Haemophilus influenzae type b (Hib), Streptococcus pneumoniae (pneumococcus), Neisseria meningitides (meningococcus) and group B streptococcus (GBS), cause a major proportion of disease in early childhood . Native PS vaccines are immunogenic and provide protection against disease in healthy adults but do not induce immunological memory . PSs are T-cell-independent antigens and do not elicit antibodies in infants and young children, but by conjugating PS to proteins they become T-cell dependent and immunogenic at an early age . Despite excellent efficacy of PS-protein conjugate vaccines against invasive disease, protection against mucosal infections such as pneumococcal otitis media has been less efficacious . Circulating PS-specific antibodies may protect against infections at mucosal sites, but mucosal immunoglobulin A antibodies may also contribute significantly to protection against mucosal infections . Mucosal immunization of experimental animals with conjugate vaccines against Hib, pneumococcus, meningococcus and GBS induces systemic and mucosal immune responses, which provide protection against carriage, otitis media and invasive disease in a variety of challenge models, providing new means for protection against encapsulated bacteria . In addition, mucosal immunization of neonatal mice with a pneumococcal conjugate and the nontoxic adjuvant LT-K63 has been superior to parenteral immunization in eliciting protective antibodies and PS-specific memory, and thus circumventing the limitations of antibody responses to PS that are responsible for enhanced susceptibility of neonates and infants to infections caused by encapsulated bacteria . Through T-cell dependent enhanced immunogenicity of PS-protein conjugate vaccines, mucosal immunization could be an attractive approach for early life immunization against encapsulated bacteria.

Int J Med Microbiol, 2003 Jun, 293(2-3), 145 - 52
Aerobic growth deficient Haemophilus influenzae mutants are non-virulent: implications on metabolism; Herbert M et al.; We investigated aerobic metabolism in Haemophilus influenzae to better understand its essential physiological growth pathways . We describe the isolation and characterization of transposon insertions leading to knockout mutations in lpdA, encoding dihydrolipoamide dehydrogenase . H . influenzae Rd lpdA::Tn10d-cat mutants were unable to grow aerobically and an H . influenzae type b lpdA::Tn10d-cat mutant was significantly attenuated in an infant rat infection model . Since LpdA is a functional subunit of both pyruvate dehydrogenase (aceEF) and alpha-ketoglutarate dehydrogenase (sucAB) the phenotype of the lpdA mutant was further explored by creating separate knockout mutants in the sucAB and aceEF loci . DeltaaceEF and deltasucAB mutants were both significantly attenuated in virulence in the infant rat, but only the sucAB mutant was able to grow aerobically . We therefore conclude that the ability for aerobic growth is critical for invasive disease, and furthermore that a TCA cycle enzyme, alpha-ketoglutarate dehydrogenase, appears to contribute a key metabolic function in vivo, but is not required for growth under laboratory conditions.

J Chemother, 2003 Jun, 15(3), 226 - 34
Antibacterial resistance in Streptococcus pneumoniae and Haemophilus influenzae from Italy and Spain: data from the PROTEKT surveillance study, 1999-2000; Schito AM et al.; Antibacterial resistance was evaluated among Streptococcus pneumoniae (n=252) and Haemophilus influenzae (n=202) from two centres in Spain (Barcelona and Madrid) and two centres in Italy (Genoa and Catania) collected during 1999-2000 as part of the ongoing PROTEKT (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin) international surveillance program . Pneumococcal nonsusceptibility to penicillin G was found to be considerably higher in Spain (53.4%) than in Italy (15.1%), whereas erythromycin A resistance was higher in Italy (42.9%) than in Spain (28.6%) . Among macrolide-resistant isolates investigated for resistance genes, the prevalence of mefA was higher among isolates from Italy (20/51, 39.2%) than among Spanish isolates (2/38, 5.3%) . All other macrolide-resistant isolates possessed ermB . Telithromycin possessed good anti-pneumococcal activity against isolates from both countries (MIC90 0.03 mg/L {Spain}; 0.25 mg/L {Italy}), irrespective of resistance to other antibacterials . Beta-lactamase production among H . influenzae was low: Spain, 10.9%; Italy, 1.8% . With the exception of ampicillin and co-trimoxazole, all H . influenzae isolates were highly susceptible to the antibacterials tested, and all were inhibited by telithromycin at a concentration of < or = 2 mg/L . The findings of PROTEKT 1999-2000 highlight the importance of local resistance patterns in guiding the choice of empirical antibacterials for community-acquired respiratory tract infections.

Pediatr Infect Dis J, 2003 Jul, 22(7), 589 - 92
Influence of child care on nasopharyngeal carriage of Streptococcus pneumoniae and Haemophilus influenzae; Dunais B et al.; BACKGROUND: Children cared for by a child minder (CM) should be less exposed to upper respiratory tract infections than those in group day care (GDC) and therefore to antibiotic treatment . Thus fewer CM children should carry resistant bacteria . To test this hypothesis nasopharyngeal carriage of Streptococcus pneumoniae (SP) and Haemophilus influenzae (HI) and exposure to recent antibiotic treatment were investigated among children in both types of care settings in the Alpes Maritimes (France) between November 1999 and March 2000 . METHODS AND POPULATION: A two stage cluster sample of children attending group day care or cared for by a child minder was selected . Nasopharyngeal samples were cultured for SP and HI . Penicillin susceptibility was tested by disk diffusion and E-test and beta-lactamase production . RESULTS: We sampled 235 children in the CM group and 298 in the GDC group who were ages 6 to 36 months . Age and sex distribution were similar in both groups . S . pneumoniae was isolated in 80 children in the CM group (34.0%) and in 163 (54.7%) children in GDC (P < 10-6) . Proportions of non-penicillin susceptible (NPSP) were 52.5 and 55.8%, respectively (P = 0.6) . H . influenzae was present in 37.2% of children in GDC vs . 23.8% in the CM group (P < 0.001) . Proportions of beta-lactamase-positive HI (HIBL+) were 40.2% vs . 46.4%, respectively (P = 0.4) . Antibiotic exposure during the previous 3 months occurred in 41.3% of children in GDC and in 47.4% in the CM group (P = 0.16) . There was no association between antibiotic use and carriage of NPSP or HIBL+ strains . CONCLUSION: SP and HI carriage rates were significantly lower among children in the CM group than in GDC . The proportion of NPSP and HIBL+ was similar in both groups, and comparable patterns of antibiotic use were observed . Continued efforts must concentrate on parental education and enforcement of recommendations for management of pediatric upper respiratory tract infections.

J Med Microbiol, 2003 Aug, 52(Pt 8), 693 - 6
Outer-membrane-protein subtypes of Haemophilus influenzae isolates from North India; Kaur R et al.; Haemophilus influenzae serotype b and non-typable isolates from blood, cerebrospinal fluid, sputum and throat swabs of patients and carriers in North India were analysed by outer-membrane protein (OMP) profiling . OMP analysis could differentiate the samples into 18 different subtypes . The non-typable isolates were more variable than the serotype b samples . OMP subtypes 1-6 were found only among the serotype b isolates and subtypes 7-18 among the non-typable isolates, while subtypes 2 and 8 were exhibited by both . The OMP profiles of isolates from blood, cerebrospinal fluid and sputum are in complete agreement with their ribotypes and RAPD fingerprints . The present study demonstrates for the first time the subtyping of Indian H . influenzae isolates by an easy and less-expensive method that is applicable to developing countries like India.

J Bacteriol, 2003 Aug, 185(15), 4593 - 602
Initial proteome analysis of model microorganism Haemophilus influenzae strain Rd KW20; Kolker E et al.; The proteome of Haemophilus influenzae strain Rd KW20 was analyzed by liquid chromatography (LC) coupled with ion trap tandem mass spectrometry (MS/MS) . This approach does not require a gel electrophoresis step and provides a rapidly developed snapshot of the proteome . In order to gain insight into the central metabolism of H . influenzae, cells were grown microaerobically and anaerobically in a rich medium and soluble and membrane proteins of strain Rd KW20 were proteolyzed with trypsin and directly examined by LC-MS/MS . Several different experimental and computational approaches were utilized to optimize the proteome coverage and to ensure statistically valid protein identification . Approximately 25% of all predicted proteins (open reading frames) of H . influenzae strain Rd KW20 were identified with high confidence, as their component peptides were unambiguously assigned to tandem mass spectra . Approximately 80% of the predicted ribosomal proteins were identified with high confidence, compared to the 33% of the predicted ribosomal proteins detected by previous two-dimensional gel electrophoresis studies . The results obtained in this study are generally consistent with those obtained from computational genome analysis, two-dimensional gel electrophoresis, and whole-genome transposon mutagenesis studies . At least 15 genes originally annotated as conserved hypothetical were found to encode expressed proteins . Two more proteins, previously annotated as predicted coding regions, were detected with high confidence; these proteins also have close homologs in related bacteria . The direct proteomics approach to studying protein expression in vivo reported here is a powerful method that is applicable to proteome analysis of any (micro)organism.

J Antimicrob Chemother, 2003 Aug, 52(2), 229 - 46 Epub 2003 Jul 15.
The Alexander Project 1998-2000: susceptibility of pathogens isolated from community-acquired respiratory tract infection to commonly used antimicrobial agents; Jacobs MR et al.; OBJECTIVES: The Alexander Project is a continuing surveillance study, begun in 1992, examining the susceptibility of pathogens involved in adult community-acquired respiratory tract infections (CARTI) to a range of antimicrobial agents . MATERIALS AND METHODS: This study tested the susceptibility of isolates of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis collected between 1998 and 2000 to 23 antimicrobials . Minimum inhibitory concentrations of agents were determined using the broth microdilution method and interpreted according to NCCLS and pharmacokinetic/pharmacodynamic (PK/PD) breakpoints . RESULTS: In total, 8882 isolates of S . pneumoniae, 8523 isolates of H . influenzae and 874 isolates of M . catarrhalis were collected during 1998-2000 from centres in 26 countries . The world-wide prevalence of penicillin resistance (penicillin MICs > or = 2 mg/l) in isolates of S . pneumoniae was 18.2% over the study period, and the prevalence of macrolide resistance (erythromycin MICs > or = 1 mg/l) in this pathogen was 24.6% . Over the study period, macrolide resistance exceeded penicillin resistance in 19 of the 26 countries included in the study . Of the non-fluoroquinolone agents, the only oral agents to which over 90% of S . pneumoniae isolates were susceptible at both NCCLS and PK/PD breakpoints were amoxicillin (95.1%) and co-amoxiclav (95.5-97.9%) . The prevalence of fluoroquinolone-resistant S . pneumoniae (ofloxacin MICs > or = 8 mg/l) was 1.1% . Gemifloxacin was the most potent fluoroquinolone tested against S . pneumoniae (99.9% susceptible) . In isolates of H . influenzae, beta-lactamase production was 16.9%, whereas the prevalence of beta-lactamase-negative, ampicillin-resistant strains was low (0.2%) . beta-Lactamase production in M . catarrhalis world-wide remained high over the period studied (92.1%) . Using PK/PD breakpoints, the most active non-fluoroquinolone agents against H . influenzae were ceftriaxone (100% susceptible), cefixime (99.8%) and co-amoxiclav (98.1-99.6%) . Co-amoxiclav, cefdinir and cefixime (100%) were the most active beta-lactams against M . catarrhalis . Both H . influenzae and M . catarrhalis were highly susceptible to the fluoroquinolones . CONCLUSIONS: These data demonstrate the continued evolution of and geographical variation in bacterial resistance and highlight the need for appropriate prescribing of antimicrobials in CARTI, using agents with adequate activity, based on local susceptibility profiles and PK/PD parameters.

Am J Rhinol, 2003 May-Jun, 17(3), 143 - 7
Bacteriology of chronic maxillary sinusitis and normal maxillary sinuses: using culture and multiplex polymerase chain reaction; Kalcioglu MT et al.; BACKGROUND: Although many investigations have been performed on bacteriology of chronic sinusitis and normal sinuses, there still is much discussion . Also a new bacterial agent, Alloiococcus otitidis determined in the nasopharynx and middle ear specimens can be thought as a causative agent of sinusitis . METHODS: The bacteriology of chronic maxillary sinusitis and maxillary sinuses with normal radiogram and endoscopic findings were studied by culture methods for aerobic and anaerobic bacteria . Multiplex polymerase chain reaction (PCR) was used to investigate four bacteria in study and control groups . There were 27 specimens in the study group and 28 specimens in the control group . RESULTS: In the study group, the bacteria commonly isolated were Staphylococcus aureus (11.1%), alpha-hemolytic streptococci (11.1%), Streptococcus pneumoniae (11.1%), Haemophilus influenzae (7.4%), coagulase-negative staphylococci (7.4%), and anaerobes (33.3%) . Coagulase-negative staphylococci (14.3%), alpha-hemolytic streptococci (10.7%), and anaerobes (35.7%) were isolated also in the control group . PCR was used to investigate S . pneumoniae, H . influenzae, Moraxella catarrhalis, and A . otitidis in the study and control groups . None of these bacteria was determined in the control group whereas detection rates of these bacteria in the study group were 11.1, 11.1, 3.7, and 7.4%, respectively . It should be considered that PCR yielded faint amplification band for A . otitidis . CONCLUSION: Using multiplex PCR can help to increase detection rates of bacterial etiology . Healthy sinuses are not sterile . A . otitidis may be one of the pathogens causing sinusitis.

Vestn Ross Akad Med Nauk, 2003, (6), 9 - 12
{Associated infections in acute bronchopulmonary infections in children}; Lykova EA et al.; A total of 189 children with bacterial complications of the acute respiratory viral infection (ARVI)--primarily with pneumonia and bronchitis--were dynamically examined for typical and atypical pneumotropic causative agents of the infection process (Mycoplasma pneumoniae, Chlamydia spp., Streptococcus pneumoniae, Haemophilus influenzae, Pneumocystis carini, and Citomegalovirus) . A high frequency rate of the associative infection involving mycoplasmas and pneumocysts was registered (45-50%); it was lower in the cases involving Chlamydias, hemophilic bacteria, pneumococcus, and cytomegalovirus--up to 25-30% . No sharp difference was found between the indices of an infection degree and those of an active clinical infectious process involving the same pneumotropic agent: the biggest difference was observed in Chlamydia infections (9.4%) and the lowest one--in mycoplasma infections (3%) . A dynamic comparison of different classes of immunoglobulins revealed that, in acute bronchitis and pneumonias, the Chlamydia and cytomegalovirus infections are, primarily, of the persistent nature; the hemophilic and pneumocystic infections are of a mixed nature; and the pneumococcus one is of the acute nature . The Mycoplasma infection, which is more often encountered in pre-school children, is of the primary type with a trend towards a prolonged clinical course . All pneumonias had a typical clinical course; the clinical picture was compared in 128 patients with the etiological factor (including a description of characteristic symptoms).

Respirology, 2003 Mar, 8(1), 83 - 9
Lower respiratory tract infection in hospitalized children; Yin CC et al.; OBJECTIVE: The aim of the present study was to investigate the aetiology and antibiotic-resistance patterns of community-acquired lower respiratory tract infection (LRTI) in 1999 and compare it with data from 1995 and 1988 . METHODOLOGY: A prospective observational study of LRTI in hospitalized children at KK Women's & Children's Hospital, Singapore, was undertaken . RESULTS: A positive isolate was found in 58% of patients (671/1158), comprising viruses (n = 477, 41.2%), non-type B Haemophilus influenzae (n = 101, 8.7%), Streptococcus pneumoniae (n = 66, 5.7%), Mycoplasma pneumoniae (n = 92, 8%), Moraxella catarrhalis (n = 19, 1.6%) and other bacteria (n = 19, 1.6%) . Mixed virus-bacteria (n = 104, 9%) infections were comprised mostly of virus-H . influenzae combinations . In 1999, S . pneumoniae resistance rates were penicillin 44.6% (17%, 1995), amoxycillin 3% (18%, 1995; MIC 0.5-2 microg/mL reclassified as susceptible in 1999), erythromycin 55% (30%, 1995), trimethoprim-sulfamethoxazole (TMP/SMX) 60% (23%, 1995) . H . influenzaeresistance rates were amoxycillin 26.7% (38%, 1995), erythromycin 99% (37%, 1995), TMP/SMX 98% (37%, 1995) . There were 15 cases of empyema of which seven were proven S . pneumoniae and there was one pneumococcal death (0.08%) . CONCLUSIONS: Pneumococcal pneumonia needs to be treated aggressively due to its high morbidity . Amoxycillin still remains useful for treating pneumococcus despite an increasing resistance to penicillin, erythromycin and TMP/SMX . Judicious use of antibiotics is needed to curb the increasing rate of antibiotic-resistance.

Proc Natl Acad Sci U S A, 2003 Jul 22, 100(15), 8898 - 903 Epub 2003 Jul 10.
Host-derived sialic acid is incorporated into Haemophilus influenzae lipopolysaccharide and is a major virulence factor in experimental otitis media; Bouchet V et al.; Otitis media, a common and often recurrent bacterial infection of childhood, is a major reason for physician visits and the prescription of antimicrobials . Haemophilus influenzae is the cause of approximately 20% of episodes of bacterial otitis media, but most strains lack the capsule, a factor known to play a critical role in the virulence of strains causing invasive H . influenzae disease . Here we show that in capsule-deficient (nontypeable) strains, sialic acid, a terminal residue of the core sugars of H . influenzae lipopolysaccharide (LPS), is a critical virulence factor in the pathogenesis of experimental otitis media in chinchillas . We used five epidemiologically distinct H . influenzae isolates, representative of the genetic diversity of strains causing otitis media, to inoculate the middle ear of chinchillas . All animals developed acute bacterial otitis media that persisted for up to 3 wk, whereas isogenic sialic acid-deficient mutants (disrupted sialyltransferase or CMP-acetylneuraminic acid synthetase genes) were profoundly attenuated . MS analysis indicated that WT bacteria used to inoculate animals lacked any sialylated LPS glycoforms . In contrast, LPS of ex vivo organisms recovered from chinchilla middle ear exudates was sialylated . We conclude that sialylated LPS glycoforms play a key role in pathogenicity of nontypeable H . influenzae and depend on scavenging the essential precursors from the host during the infection.

Respir Med, 2003 Jul, 97(7), 770 - 7
Bacterial infections in patients requiring admission for an acute exacerbation of COPD; a 1-year prospective study; Groenewegen KH et al.; STUDY OBJECTIVE: To investigate the frequency of respiratory bacterial infections in hospitalized patients, admitted with an acute exacerbation of chronic obstructive pulmonary disease (COPD), to identify the responsible pathogens by sputum culture and to assess patient characteristics in relation to sputum culture results . METHODS: We prospectively evaluated clinical data and sputum culture results of 171 patients, admitted to the pulmonology department of the University Hospital Maastricht with an acute exacerbation of COPD from 1st January 1999 until 31st December 1999 . RESULTS: Eighty-five patients (50%) had positive sputum cultures, indicating the presence of bacterial infection . Pathogens most frequently isolated were: Haemophilus influenzae (45%), Streptococcus pneumoniae (27%), and Pseudomonas aeruginosa (15%) . Patients with more severely compromised lung function had a higher incidence of bacterial infections (P = 0.026) . There were no significant differences in age, lung function parameters, blood gas results and length of hospital stay between patients with and without bacterial infection . There were no correlations between the type of bacteria isolated and clinical characteristics . CONCLUSION: Incidence of bacterial infection during acute exacerbations of COPD is about 50% . Patients with and without bacterial infection are not different in clinical characteristics or in outcome parameters . Patients with lower FEV1 have a higher incidence of bacterial infections, but there is no difference in the type of bacterial infection . In the future, the pathogenic role of bacterial infection in exacerbations of COPD should be further investigated, especially the role of bacterial infection in relation to local and systemic inflammation.

Avian Pathol, 2003 Jun, 32(3), 265 - 9
Efficacy of a new tetravalent coryza vaccine against emerging variant type B strains; Jacobs AA et al.; Outbreaks of infectious coryza have been reported in vaccinated flocks in different countries, indicating that new serotype(s) of Haemophilus paragallinarum may have evolved . Several field isolates from vaccinated flocks in the US, Ecuador, Argentina and Zimbabwe were examined and, apart from one serotype C strain, all were typed as serotype B . An inactivated commercial trivalent vaccine, containing serotypes A, B and C, protected against challenge with the serotype C isolate but protection against challenge with serotype B isolates was weaker, suggesting that they might represent a new variant immunotype . An experimental tetravalent oil adjuvant vaccine, containing one of the serotype B isolates, appeared immunogenic against all isolates after one vaccination . Its efficacy and safety were further tested in layer chickens housed under field conditions . Chickens were vaccinated at 8 and 16 weeks of age while controls were unvaccinated . Vaccinates and controls were challenged with type A, B, C and variant type B at 25, 45 or 65 weeks of age . There was good protection (P<0.05) against all four immunotypes after all challenges . No systemic reactions were observed and local reactions were similar to those found with the commercial trivalent vaccine . The tetravalent vaccine may therefore be a good choice for control of new field isolates.

Vaccine, 2003 Jul 28, 21(24), 3455 - 9
Maternal immunization with Haemophilus influenzae type b vaccines in different populations; Englund JA et al.; Haemophilus influenzae type b (Hib) vaccines provide an excellent model for maternal immunization because effective vaccines are readily available and the vaccines are safe and reliable, and markers of efficacy have been established and standardized . Studies of polysaccharide and conjugate Hib vaccines administered to pregnant women and women of childbearing ages are reviewed in this paper . The type of vaccine has been shown to be important in increasing transplacental passage of maternal antibody . The timing of vaccine during pregnancy is also important in the transfer of this antibody . The total amount of IgG antibody in the mother, as well as the isotype class and subclass of IgG antibody, influences the final levels of antibody in the neonate . Placental integrity has been shown to be important in the active transport of antibody from mother to fetus . The impact of increased levels of Hib antibody in infants at the time of primary immunization with Hib does not appear to interfere with vaccine efficacy, although higher antibody levels in infants at the time of immunization may result in lower total antibody levels following all doses of vaccine . Principles observed in these studies have potential application against other important neonatal pathogens.

Vaccine, 2003 Jul 28, 21(24), 3451 - 4
Maternal immunization with pneumococcal polysaccharide vaccine in the Philippines; Quiambao BP et al.; A randomized, controlled study was conducted to evaluate the immunogenicity and reactogenicity of the 23-valent pneumococcal (Pnc) polysaccharide (PS) vaccine among pregnant women and to ascertain the transfer of anti-Pnc antibody (Ab) from mother to infant . One hundred and sixty women received either one dose of Pnc PS vaccine, Haemophilus influenzae type b conjugate vaccine and tetanus toxoid (TT) (Pnc vaccine group, N=106) or TT only (control group, N=54) . Sera were obtained from all mothers prior to vaccination and 4 weeks after from the vaccinated group . Cord blood was obtained in 75% of deliveries . Anti-Pnc Ab for serotypes 1, 5, 6B, 14, 18C and 19F was determined using enzyme immunoassay . The Pnc vaccine and control groups were comparable in terms of age, parity, gravidity, prior doses of TT, and pre-vaccination geometric mean concentration (GMC in microg/ml) of anti-Pnc Ab . Between 66 and 87% of the mothers had type-specific Ab prior to vaccination . There was a significant rise in anti-Pnc Ab (varying from 3.3- to 9.1-fold for the individual serotypes) between the pre and post-vaccination samples . Adverse reactions were mild and required no treatment . The level of anti-Pnc Ab in cord blood was significantly lower in the control group compared to the Pnc vaccine group . Vaccination of pregnant women with Pnc Ps vaccine induces good immune response and Ab can be transferred to their infants via cord blood thus providing enhanced protection.

Vaccine, 2003 Jul 28, 21(24), 3393 - 7
The influence of maternal immunization on light chain response to Haemophilus influenzae type b vaccine; Nahm MH et al.; The repertoire of human antibodies to Haemophilus influenzae type b polysaccharide (Hib PS) has been extensively characterized . In response to Hib PS vaccines, both IgG1 and IgG2 antibodies are produced . Based on the Vkappa gene used for the antibodies, heamophilus antibodies can be divided into A2 and non-A2 antibodies . A2 antibodies account for the majority of heamophilus antibodies, are generally more functional, and are specific for heamophilus polysaccharide . In contrast, non-A2 antibodies cross-react with Escherichia coli K100 . Studies of placental transfer showed that IgG1 antibodies cross the placenta better than IgG2 antibodies and the A2 antibodies better than non-A2 antibodies . Babies born to mothers who were immunized with a heamophilus vaccine had a high level of antibodies to Hib PS but the antibody level decreased during the first 6 months . The children also responded to a heamophilus conjugate vaccine.

Clin Microbiol Infect, 2003 May, 9(5), 449 - 52
Evaluation of the WIDER I system for antimicrobial susceptibility testing of clinical isolates of Haemophilus influenzae and Streptococcus pneumoniae; Fernandez-Cuenca F et al.; The aim of this study was to evaluate the WIDER I system for susceptibility testing of Haemophilus influenzae and Streptococcus pneumoniae . MICs of 12 antimicrobials against 42 H . influenzae and 58 S . pneumoniae strains were determined using 1W MIC panels and compared with those obtained by microdilution . Overall essential agreements were >99% . Very major errors were not detected . Major errors occurred with ampicillin (1.7% H . influenzae) . Minor errors were 2.3% (amoxicillin-clavulanate, cefuroxime, chloramphenicol), 7.1% (ampicillin) and 16.7% (clarithromycin) for H . influenzae, and 1.7% (chloramphenicol, erythromycin, meropenem), 3.4% (amoxicillin-clavulanate, cefuroxime, tetracycline) and 8.6% (levofloxacin) for S . pneumoniae . The WIDER I system is a reliable method for susceptibility testing of H . influenzae and S . pneumoniae.

Clin Microbiol Infect, 2003 May, 9(5), 431 - 6
Antimicrobial susceptibility of 840 clinical isolates of Haemophilus influenzae collected in four European countries in 2000-2001; Blosser-Middleton R et al.; In 2000-2001, 840 clinical isolates of Haemophilus influenzae were collected from laboratories in France, Germany, Italy and Spain (210 isolates/country) . Beta-Lactamase production among the isolates varied considerably by country, ranging from 8.1% in Germany to 34.8% in France . H . influenzae from patients <or=4 years old showed the highest prevalence of beta-lactamase production (23.2%), compared with isolates from patients aged 5-17 years (17.8%) and >or=18 years (16.5%) . All isolates were susceptible to amoxicillin-clavulanate, ciprofloxacin and levofloxacin; 99.6% and 98.9% of isolates were susceptible to azithromycin and cefuroxime, respectively . Among the macrolides tested, azithromycin (MIC90, 2 mg/L) was eight-fold more potent than clarithromycin (MIC90, 16 mg/L) and roxithromycin (MIC90, 16 mg/L) . Despite variations in beta-lactamase production between different countries, > 99% of all isolates were susceptible to amoxicillin-clavulanate, ciprofloxacin, levofloxacin, and azithromycin.

Infez Med, 1997, 5(1), 52 - 6
{Septic arthritis in the setting of HIV disease . A case report and literature review}; Manfredi R et al.; A case of septic arthritis due to Haemophilus influenzae is described in a HIV-infected patient . Even though complicated by large effusion and extensive subcutaneous involvement, the clinical picture showed a favourable outcome after prolonged ceftriaxone treatment, leading to complete cure . The case report is discussed with respect to both other bacterial complications caused by H . influenzae in the setting of HIV disease, and the broad aetiological spectrum of HIV-associated acute arthritis . In particular, no other cases of H . influenzae purulent arthritis have been described until now in patients with HIV disease, at our best knowledge.

Eur J Biochem, 2003 Jul, 270(14), 2979 - 91
Structural characterization of a novel branching pattern in the lipopolysaccharide from nontypeable Haemophilus influenzae; Mansson M et al.; Structural analysis of the lipopolysaccharide (LPS) from nontypeable Haemophilus influenzae strain 981 has been achieved using NMR spectroscopy and ESI-MS on O-deacylated LPS and core oligosaccharide (OS) material as well as by ESI-MSn on permethylated dephosphorylated OS . A heterogeneous glycoform population was identified, resulting from the variable length of the OS branches attached to the glucose residue in the common structural element of H . influenzae LPS, l-alpha-d-Hepp-(1-->2)-{PEtn-->6}-l-alpha-d-Hepp-(1-->3)-{beta-d-Glcxp-(1-->4)}-l-alpha-d-Hepp-(1-->5)-{PPEtn-->4}-alpha-Kdop-(2-->6)-Lipid A . Notably, the O-6 position of the beta-d-Glcp residue was either substituted by PCho or the disaccharide branch beta-d-Galp-(1-->4)-d-alpha-d-Hepp, while the O-4 position was substituted by the globotetraose unit, beta-d-GalpNAc-(1-->3)-alpha-d-Galp-(1-->4)-beta-d-Galp-(1-->4)-beta-d-Glcp, or sequentially truncated versions thereof . This is the first time a branching sugar residue has been reported in the outer-core region of H . influenzae LPS . Additionally, a PEtn group was identified at O-3 of the distal heptose residue in the inner-core.

Immunogenetics, 2003 Aug, 55(5), 336 - 8 Epub 2003 Jul 04.
IGH V3-23*01 and its allele V3-23*03 differ in their capacity to form the canonical human antibody combining site specific for the capsular polysaccharide of Haemophilus influenzae type b; Liu L et al.; The IGH V3-23*01 gene is used in the formation of the canonical combining site which dominates the human antibody repertoire to the Haemophilus influenzae type b (Hib) polysaccharide (PS) . An allele of the human IGH V3-23*01 gene, known as V3-23*03, differs from V3-23*01 in nine bases, eight of which are located in the second complementarity determining region . These eight differences encode five amino acid substitutions . In this study we investigated whether the V3-23*03 sequence polymorphism affected Hib PS binding . We constructed two Fab fragments that had the canonical Hib PS combining site VH-VL configuration but that had either V3-23*01 or V3-23*03 . Radioantigen binding assay showed that on a concentration basis the V3-23*03 Fab was 20-fold more effective in binding Hib PS than the V3-23*01 Fab . The V3-23*03 Fab was 4-fold more effective than the V3-23*01 Fab in mediating facilitated bactericidal activity against Hib organisms . These findings identify a functional consequence of V3-23 allelism, and suggest that utilization of the V3-23*03 gene in the human Hib PS repertoire would generate canonical antibodies with higher affinity and protective efficacy than canonical antibodies utilizing V3-23*01 . Thus, IGH V gene allelic variation has the potential to impact the generation of protective immunity to Hib.

J Clin Microbiol, 2003 Jul, 41(7), 3064 - 9
Invasive disease due to nontypeable Haemophilus influenzae among children in Arkansas; O'Neill JM et al.; In this study, we reviewed cases of invasive disease due to nontypeable Haemophilus influenzae among children hospitalized at Arkansas Children's Hospital from 1993 to 2001 . A total of 28 cases were examined, including 21 associated with bacteremia and 4 associated with meningitis . Of the patients examined, 86% were </=4 years of age, and 68% had underlying medical conditions . Characterization of the bacterial isolates by multilocus sequence type genotyping revealed significant overall genetic diversity, similar to the diversity in the general population structure for nontypeable H . influenzae . However, four separate pairs of isolates were closely related genetically, a relationship confirmed by pulsed-field gel electrophoresis and Southern hybridization studies using probes for the major H . influenzae adhesin genes . These results suggest that selected strains of nontypeable H . influenzae may have more invasive potential, especially in young children and patients with underlying medical conditions . At this point, the specific factors that contribute to enhanced virulence remain unclear.

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