Haemophilus

Int J Antimicrob Agents, 2001 Jun, 17(6), 457 - 64
Susceptibility of Canadian isolates of Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae to oral antimicrobial agents; Blondeau JM et al.; We measured the susceptibility of Canadian isolates of three respiratory tract pathogens (Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae) to several currently approved antimicrobial agents by two different methods . We also measured the susceptibility of isolates to seven fluoroquinolones . Beta-lactamase was produced by 123/566 (21.7%) of H . influenzae isolates compared with 178/200 (89%) of M . catarrhalis isolates . For S . pneumoniae 83/374 (22.2%) isolates were penicillin resistant and of these 2.1% (8/374) showed high level resistance (MIC > or = 2 mg/l) . Regardless of methodology, all fluoroquinolones were highly active against H . influenzae (MIC(90) < or = 0.031 mg/l) and M . catarrhalis (MIC(90) < or = 0.064 mg/l) isolates . Susceptibility of H . influenzae to cefuroxime and amoxycillin/clavulanic acid was 99-100% whereas 84-85.5% were susceptible to cefaclor and cefprozil . Azithromycin susceptibility ranged from 82.6 to 99.2% depending on the method . M . catarrhalis isolates were uniformly susceptible to all agents tested except amoxycillin . Cross-resistance in S . pneumoniae to all non-quinolone agents was concurrent with increasing penicillin resistance as shown by increasing MIC90 values . For the fluoroquinolones tested, the rank order of potency based on MIC(90) values was as follows: gemifloxacin (0.031-0.063 mg/l), trovafloxacin (0.125 mg/l), moxifloxacin (0.125-0.25 mg/l), grepafloxacin (0.125-0.25 mg/l), gatifloxacin (0.5 mg/l), levofloxacin (1 mg/l) and ciprofloxacin (2 mg/l) . Our study confirms either a high or increasing prevalence of antimicrobial resistant respiratory pathogens in Canada and also compares the new and old fluoroquinolones and their potential role as therapy for community-acquired infections . The prevalence of beta-lactamase positive H . influenzae may have decreased from levels reported in previous studies.

Int J Antimicrob Agents, 2001 Jun, 17(6), 451 - 5
Comparative antimicrobial activity of ABT-773, a novel ketolide, tested against drug-resistant Gram-positive cocci and Haemophilus influenzae; Rospide MF et al.; The antimicrobial activity of ABT-773, a novel ketolide, was tested against 618 Gram-positive strains collected from various surveillance programmes between 1997 and 2000 . ABT-773 has potent activity against Streptococcus pneumoniae (MIC90, < or = 0.03-0.12 mg/l), beta-haemolytic streptococci (MIC90, < or = 0.03 mg/l) and viridans group streptococci (MIC90, < or = 0.03 mg/l), including erythromycin-resistant strains . In contrast, ABT-773 was less active against erythromycin-resistant Staphylococcus aureus (31% susceptible at < or = 0.25 mg/l), coagulase-negative staphylococci (41% susceptible) and enterococci (30% susceptible) . Haemophilus influenzae (MIC90, 4 mg/l) was less inhibited by the two ketolides tested, and ABT-773 was generally two- to fourfold more potent than telithromycin . The ketolides appear to have potential clinical use against some Gram-positive species resistant to macrolides.

J Bacteriol, 2001 Jul, 183(13), 4004 - 11
Global versus local regulatory roles for Lrp-related proteins: Haemophilus influenzae as a case study; Friedberg D et al.; Lrp (leucine-responsive regulatory protein) plays a global regulatory role in Escherichia coli, affecting expression of dozens of operons . Numerous lrp-related genes have been identified in different bacteria and archaea, including asnC, an E . coli gene that was the first reported member of this family . Pairwise comparisons of amino acid sequences of the corresponding proteins shows an average sequence identity of only 29% for the vast majority of comparisons . By contrast, Lrp-related proteins from enteric bacteria show more than 97% amino acid identity . Is the global regulatory role associated with E . coli Lrp limited to enteric bacteria? To probe this question we investigated LrfB, an Lrp-related protein from Haemophilus influenzae that shares 75% sequence identity with E . coli Lrp (highest sequence identity among 42 sequences compared) . A strain of H . influenzae having an lrfB null allele grew at the wild-type growth rate but with a filamentous morphology . A comparison of two-dimensional (2D) electrophoretic patterns of proteins from parent and mutant strains showed only two differences (comparable studies with lrp(+) and lrp E . coli strains by others showed 20 differences) . The abundance of LrfB in H . influenzae, estimated by Western blotting experiments, was about 130 dimers per cell (compared to 3,000 dimers per E . coli cell) . LrfB expressed in E . coli replaced Lrp as a repressor of the lrp gene but acted only to a limited extent as an activator of the ilvIH operon . Thus, although LrfB resembles Lrp sufficiently to perform some of its functions, its low abundance is consonant with a more local role in regulating but a few genes, a view consistent with the results of the 2D electrophoretic analysis . We speculate that an Lrp having a global regulatory role evolved to help enteric bacteria adapt to their ecological niches and that it is unlikely that Lrp-related proteins in other organisms have a broad regulatory function.

J Bacteriol, 2001 Jul, 183(13), 3974 - 81
NadN and e (P4) are essential for utilization of NAD and nicotinamide mononucleotide but not nicotinamide riboside in Haemophilus influenzae; Kemmer G et al.; Haemophilus influenzae has an absolute requirement for NAD (factor V) because it lacks almost all the biosynthetic enzymes necessary for the de novo synthesis of that cofactor . Factor V can be provided as either nicotinamide adenosine dinucleotide (NAD), nicotinamide mononucleotide (NMN), or nicotinamide riboside (NR) in vitro, but little is known about the source or the mechanism of uptake of these substrates in vivo . As shown by us earlier, at least two gene products are involved in the uptake of NAD, the outer membrane lipoprotein e (P4), which has phosphatase activity and is encoded by hel, and a periplasmic NAD nucleotidase, encoded by nadN . It has also been observed that the latter gene product is essential for H . influenzae growth on media supplemented with NAD . In this report, we describe the functions and substrates of these two proteins as they act together in an NAD utilization pathway . Data are provided which indicate that NadN harbors not only NAD pyrophosphatase but also NMN 5'-nucleotidase activity . The e (P4) protein is also shown to have NMN 5'-nucleotidase activity, recognizing NMN as a substrate and releasing NR as its product . Insertion mutants of nadN or deletion and site-directed mutants of hel had attenuated growth and a reduced uptake phenotype when NMN served as substrate . A hel and nadN double mutant was only able to grow in the presence of NR, whereas no uptake of NMN was observed.

Vaccine, 2001 Jun 14, 19(27), 3645 - 51
Impact of osmolality on burning sensations during and immediately after intramuscular injection of 0.5 ml of vaccine suspensions in healthy adults; Nony P et al.; A randomised placebo controlled double-blind cross-over trial was performed on twenty healthy adults to assess the effect of osmolality (300,600,850 and 1100 mOsm) on local tolerance of an intramuscular injection (0.5 ml) of five suspensions containing the same components as the excipients of a combined Diphtheria-Tetanus-acellular Pertussis-inactivated Poliomyelitis-Haemophilus influenzae type b paediatric vaccine (DtacP-IPV-Hib, PENTAVAC) . The results did not show any dose-effect relationship between burning or pain sensations and the different osmolalities tested . Although mild and not clinically relevant, these sensations seemed to occur more frequently following injection of an isotonic saline solution (P<0.05) . Thus, the osmolality of vaccine like suspensions does not appear to be a potential cause of local pain or burning sensation after their administration.

Int J STD AIDS, 2001 Jul, 12(7), 419 - 22
Vaccine candidates in STD; Fletcher MA; Sexually transmitted diseases (STDs) are caused by organisms that infect the mucosal surfaces of the genitourinary tract . In spite of its public health importance, particular scientific problems have delayed the development of an STD vaccine, such as incomplete attenuation (human herpes simplex virus type 2), accentuated immunopathology (Chlamydia trachomatis), poor immunogenicity (Treponema pallidum), and broad antigenic heterogeneity (Neisseria gonorrhoeae) . Nevertheless, efforts continue with the use of protein antigens: for example, the haemolysin toxoid of Haemophilus ducreyi; the major outer membrane protein(s) of N . gonorrhoeae and C . trachomatis; the glycoprotein D of human herpes simplex virus type 2; and the proteins E6 and E7 of the human papillomavirus . It could be predicted that eventual STD vaccines (administered either for prophylaxis or for therapy) will use approaches that will include (1) live-attenuated viruses, (2) subunit proteins or inactivated whole organisms given with mucosal adjuvants or with cellular immune response adjuvants, or (3) DNA plasmids expressing the vaccine antigen.

Mem Inst Oswaldo Cruz, 2001 May, 96(4), 583 - 6
A comparative study of preservation and storage of Haemophilus influenzae; Aulet de Saab OC et al.; The aim of this study was to compare the efficacy of conservation by freezing the strains of Haemophilus influenzae at -20 degrees C and -70 degrees C . Skim milk supplemented with glucose, yeast extract and glycerol allowed highest viability of H . influenzae both at -20 degrees C and -70 degrees C from the media analyzed . Trypticase soy broth and brain heart infusion broth supplemented with glycerol, allowed excellent recovery . Use of cotton swaps as supporting material, with or without addition of cryoprotective agents, did not modify H . influenzae viability after six months of storage . Concentration of the initial inoculum positively affected viability when stored at -20 degrees C . Initial concentration did not influence survival after storage at -70 degrees C . Thawing at room temperature should not exceed 3 h as to get highest survival percentage.

Vet Microbiol, 2001 Aug 8, 81(3), 243 - 55
Phenotypic and genetic characterization of NAD-dependent Pasteurellaceae from the respiratory tract of pigs and their possible pathogenetic importance; Kielstein P et al.; Nicotinamide adenine dinucleotide (NAD)-dependent Pasteurellaceae other than Actinobacillus pleuropneumoniae and Haemophilus parasuis are frequently isolated from the respiratory tract of pigs . The taxonomic classification and relevance for pathogenicity of these bacteria deserves further attention . In the present study, 107 of these NAD-dependent isolates from the porcine respiratory tract, primarily from lungs with pathological changes, were investigated . On the basis of phenotypic criteria, such as haemolysis, urease, catalase, and indole formation as well as other fermentative activities, 50 of the isolates were assigned to Actinobacillus minor, 36 isolates to Actinobacillus porcinus and 21 isolates to Actinobacillus indolicus . However, many isolates among the three species showed fermentative activities differing from those of the respective type strain of the species . Serotyping on the basis of heat-stable polysaccharide antigens and 16 rDNA sequencing also revealed substantial heterogeneity within each of the three species although they clustered together in three distinct groups in the phylogenetic analysis . These three groups of NAD-dependent bacteria are different from, or in a borderline position, to the existing species or genera within the family Pasteurellaceae . A considerable number of isolates of these three groups were isolated in pure cultures from pneumonic lungs . Consequently, it will be necessary to critically review the opinion, that these NAD-dependent Pasteurellaceae are only "agents colonizing the mucosa" . Further, taxonomic examinations of the strains within these three groups are indispensable to testing isolates for their virulence in gnotobiotic pigs.

Biochem J, 2001 Jun 15, 356(Pt 3), 851 - 8
Overproduction, purification and novel redox properties of the dihaem cytochrome c, NapB, from Haemophilus influenzae; Brige A et al.; The napB gene of the pathogenic bacterium Haemophilus influenzae encodes a dihaem cytochrome c, the small subunit of a heterodimeric periplasmic nitrate reductase similar to those found in other bacteria . In order to obtain sufficient protein for biophysical studies, we aimed to overproduce the recombinant dihaem protein in Escherichia coli . Initial expression experiments indicated that the NapB signal peptide was not cleaved by the leader peptidase of the host organism . Apocytochrome was formed under aerobic, semi-aerobic and anaerobic growth conditions in either Luria--Bertani or minimal salts medium . The highest amounts of apo-NapB were produced in the latter medium, and the bulk was inserted into the cytoplasmic membrane . The two haem groups were covalently attached to the pre-apocytochrome only under anaerobic growth conditions, and with 2.5 mM nitrite or at least 10 mM nitrate supplemented to the minimal salts growth medium . In order to obtain holocytochrome, the gene sequence encoding mature NapB was cloned in-frame with the E . coli ompA (outer membrane protein A) signal sequence . Under anaerobic conditions, NapB was secreted into the periplasmic space, with the OmpA signal peptide being correctly processed and with both haem c groups attached covalently . Unless expressed in the DegP-protease-deficient strain HM125, some of the recombinant NapB polypeptides were N-terminally truncated as a result of proteolytic activity . Under aerobic growth conditions, co-expression with the E . coli ccm (cytochrome c maturation) genes resulted in a higher yield of holocytochrome c . The pure recombinant NapB protein showed absorption maxima at 419, 522 and 550 nm in the reduced form . The midpoint reduction potentials of the two haem groups were determined to be -25 mV and -175 mV . These results support our hypothesis that the Nap system fulfils a nitrate-scavenging role in H . influenzae.

Clin Infect Dis, 2001 Jul 1, 33 Suppl 1, S15 - 21
Prophylactic measures in the solid-organ recipient before transplantation; Avery RK et al.; Pretransplant screening affords an important opportunity to detect and treat preexisting active infection in the solid-organ transplant recipient . In this article, pretransplant strategies for preventing infections after solid-organ transplantation are reviewed . In addition to the search for active preexisting infection in the transplant candidate, immunization remains a cornerstone of preventive practice . Because there is a suboptimal response to vaccinations in patients who are receiving immunosuppressive therapy, as well as in patients with end-stage organ dysfunction, standard immunization of the transplant candidate should be updated as early as possible in the course of the illness, including pneumococcal, influenza, and hepatitis B vaccines . Liver transplant candidates should receive hepatitis A vaccine, and children should receive Haemophilus influenzae type B conjugate vaccine . All nonimmune pretransplant patients should be considered candidates for the varicella vaccine . The management of special risk groups is discussed in detail.

Pediatrics . 2001 Jun;107(6):E90.
Impact of the change to inactivated poliovirus vaccine on the immunization status of young children in the United States: a study from pediatric research in office settings and the National Medical Association; Taylor JA et al.; OBJECTIVE: To determine whether the change from an all oral poliovirus vaccine (OPV) schedule to an inactivated poliovirus vaccine (IPV)-containing schedule has adversely affected the immunization status of young children in the United States . METHODS: Immunization data were abstracted from the medical records of children 8 to 35 months old seen consecutively for any reason in the offices of practicing pediatricians who are members of the Pediatric Research in Office Settings network of the American Academy of Pediatrics or the National Medical Association . Data on up to 120 eligible children were collected in each practice between March 1998 and January 2000 . Patients were classified as fully immunized at 8 months old if they had received 3 diphtheria-tetanus-pertussis, 2 Haemophilus influenzae type b, 2 hepatitis B, and 2 poliovirus vaccines . Study children who were >/=12 months of age at the time that data were collected were categorized as being fully immunized at 12 months if they had received the same vaccines before their first birthday . To assess the effect of type of poliovirus vaccines on these outcomes, study patients were classified as being in an IPV or OPV group based on the initial type of vaccine received . Logistic regression was used to calculate the odds ratios (ORs) and 95% confidence intervals (CIs) for IPV as a predictor of being fully immunized at 8 and 12 months of age, after adjusting for race/ethnicity of the patient, maternal education level, year of birth, and method of payment for vaccines . In addition, the effect of clustering of children within practices was accounted for by the use of generalized estimation equation techniques . RESULTS: Data were analyzed on 13 520 children from 177 practices in 42 states; 79.4% of patients were fully immunized at 8 months of age, and 88.7% of those eligible were fully immunized at 12 months of age . A total of 6910 patients (51.1%) were classified as OPV recipients, wheras 5282 (39.1%) received IPV . In addition, 1328 children (9.8%) were documented as having received poliovirus vaccine, but the particular type could not be determined . Compared with OPV recipients and after controlling for the confounding variables and the effect of clustering within practices, children in the IPV group were as likely as were OPV recipients to be fully immunized at 8 months of age (OR: 1.04; 95% CI: 0.88,1.23) . At 12 months of age, the OR for IPV as a predictor of being fully immunized was 1.08 (95% CI: 0.90,1.30) . When compared with OPV recipients, adjusted ORs for children in the undetermined poliovirus vaccine type group being fully immunized at 8 and 12 months of age were 0.84 (95% CI: 0.68,1.04) and 0.84 (95% CI: 0.67,1.07), respectively . CONCLUSIONS: The results of this national study indicate that the implementation of an IPV-containing poliovirus vaccine schedule has not had an adverse effect on the immunization status of young children who were vaccinated in the offices of practicing pediatricians.

J Antimicrob Chemother, 2001 Jun, 47(6), 801 - 10
External quality assessment of antimicrobial susceptibility testing in Europe; Snell JJ et al.; Comparability of results of antimicrobial susceptibility testing is essential for resistance surveillance studies . As different methods may be used in different countries, there may be particular problems with international comparisons of resistance rates . Data from external quality assessment (EQA) surveys participated in by laboratories from several European countries allow comparison of performance between countries . In this study, success rates with organism-antimicrobial agent combinations known to be difficult to test were examined . With penicillin resistance in pneumococci; vancomycin and high-level gentamicin resistance in enterococci; ampicillin, co-amoxiclav and chloramphenicol resistance in Haemophilus influenzae and methicillin resistance in staphylococci there were differences between countries in success rates for discrimination of resistant strains . This study suggests that differences between countries in rates of resistance for some organism-antimicrobial agent combinations should be interpreted with caution . International EQA is useful in the demonstration and clarification of such differences.

Protein Expr Purif, 2001 Jun, 22(1), 52 - 9
Subcloning, expression, purification, and characterization of Haemophilus influenzae glycerol kinase; Pawlyk AC et al.; Glycerol kinase (EC 2.7.1.30) is a bacterial sugar kinase and a member of the sugar kinase/actin/hsc-70 superfamily of enzymes . The enzyme from Escherichia coli is an allosteric regulatory enzyme whose activity is inhibited by fructose 1,6-bisphosphate (FBP) and the glucose-specific phosphocarrier of the phosphoenolpyruvate:glycose phosphotransferase system, IIA(Glc) (previously termed III(Glc)) . Comparison of its primary structure with that of the highly similar Haemophilus influenzae glycerol kinase reveals that the amino acid sequence for the binding site for FBP is conserved while the amino acid sequence for the binding site for IIA(Glc) contains differences that are predicted to prevent its inhibition . To test this hypothesis, the H . influenzae glpK gene was assembled from DNA library fragments and subcloned into pUC18 . The enzyme is expressed at high levels in E . coli . It was purified to greater than 90% homogeneity by taking advantage of its solubility behavior in a procedure that requires no column chromatography . The initial-velocity kinetic parameters of the purified enzyme are similar to those of the E . coli glycerol kinase . The H . influenzae glycerol kinase is inhibited by FBP but not by IIA(Glc), in agreement with the prediction based on sequence comparison . Sedimentation velocity experiments reveal that inhibition of HiGK by FBP is associated with oligomerization, behavior which is similar to EcGK . The possibility of utilizing mutagenesis studies to exploit the high degree of similarity of these two enzymes to elucidate the mechanism of allosteric regulation by IIA(Glc) is discussed .

Ann Emerg Med, 2001 Jun, 37(6), 703 - 10
Principles of appropriate antibiotic use for acute rhinosinusitis in adults: background; Hickner JM et al.; The following principles of appropriate antibiotic use for adults with acute rhinosinusitis apply to the diagnosis and treatment of acute maxillary and ethmoid rhinosinusitis in adults who are not immunocompromised.Most cases of acute rhinosinusitis diagnosed in ambulatory care are caused by uncomplicated viral upper respiratory tract infections . Bacterial and viral rhinosinusitis are difficult to differentiate on clinical grounds . The clinical diagnosis of acute bacterial rhinosinusitis should be reserved for patients with rhinosinusitis symptoms lasting 7 days or more who have maxillary pain or tenderness in the face or teeth (especially when unilateral) and purulent nasal secretions . Patients with rhinosinusitis symptoms that last less than 7 days are unlikely to have bacterial infection, although rarely some patients with acute bacterial rhinosinusitis present with dramatic symptoms of severe unilateral maxillary pain, swelling, and fever . Sinus radiography is not recommended for diagnosis in routine cases . Acute rhinosinusitis resolves without antibiotic treatment in most cases . Symptomatic treatment and reassurance is the preferred initial management strategy for patients with mild symptoms . Antibiotic therapy should be reserved for patients with moderately severe symptoms who meet the criteria for the clinical diagnosis of acute bacterial rhinosinusitis and for those with severe rhinosinusitis symptoms-especially those with unilateral facial pain-regardless of duration of illness . For initial treatment, the most narrow-spectrum agent active against the likely pathogens, Streptococcus pneumoniae and Haemophilus influenzae, should be used.

Curr Infect Dis Rep, 2001 Jun, 3(3), 209 - 216
The Microbiology and Management of Acute and Chronic Rhinosinusitis; Hadley JA; Although most cases of rhinosinusitis are benign, the disruption of quality of life due to disease symptoms leads patients to seek early medical care . Ongoing debates dispute the definition, bacteriology, and medical management of chronic sinusitis, while the criteria for acute sinusitis are relatively well established . Chronic rhinosinusitis remains poorly categorized, and authors differ in opinions of symptoms, time course, and bacteriology of the infections, as well as proper medical management . Recent studies from the Mayo Clinic even question the idea that chronic sinusitis is a bacterial disease and document the presence of fungal pathogens that are responsible for the inflammatory reaction and mucosal response . In general, medical management is based on physiologic principles of re-establishing natural mucociliary function and restoring proper aeration of the paranasal sinuses after an inflammatory insult . Injudicious use and indiscriminate overprescription of antibiotics has fostered the rapid development of penicillin-resistant organisms over the past two decades . Drug-resistant Streptococcus pneumonia and b-lactamase-producing Haemophilus influenzae and Moraxella catarrhalis are becoming the norm, forcing the need to consider alternatives in antibacterial management . Appropriate medical management of this common problem requires a systematic approach and consideration of adjunctive therapy . This article examines the current bacteriology of these common infections and reviews the management of acute and chronic infections.

Pharmacoeconomics, 2001, 19(4), 391 - 400
Cost-benefit analysis of a Haemophilus influenzae type b meningitis prevention programme in The Philippines; Limcangco MR et al.; BACKGROUND: Haemophilus influenzae type b (Hib) meningitis is associated with high mortality and serious sequelae in children under 5 years of age . Vaccines which can prevent this infection are available . OBJECTIVE: To evaluate the costs and benefits of a 3-dose immunisation schedule in Manila, Philippines . PERSPECTIVE: Government and societal perspectives . DESIGN AND PARTICIPANTS: A cost-benefit analysis based on a birth cohort of 100,000 children . The state of health of the cohort with and without a Hib immunisation programme was modelled over a 5-year period . A survey of medical records of patients with Hib in Manila provided data on the extent and cost of sequelae following infection . INTERVENTION: A 3-dose Hib vaccination programme given at ages 2, 3 and 4 months . RESULTS: The model predicted that vaccinating children against Hib meningitis would prevent 553 cases per year in a birth cohort of 100,000, at a cost of 56,200 Philippine pesos (PHP) {$US1,605; 1998 exchange rate} per case (base case assumptions of 90% vaccine efficacy rate, 95 per 100,000 Hib incidence rate, 85% vaccination coverage) . Results from the cost-benefit analyses indicated that the saving to the government would be around PHP39 million ($US1.11 million), and the saving to society would be PHP255 million ($US7.28 million) . CONCLUSION: There would be a positive economic benefit for the Philippine government and for the Filipino society if a Hib vaccination programme was introduced in Manila.

Microbiol Res, 2001, 156(1), 1 - 7
A novel surfactant nanoemulsion with a unique non-irritant topical antimicrobial activity against bacteria, enveloped viruses and fungi; Hamouda T et al.; A novel non-ionic surfactant nanoemulsion designated 8N8 has been tested for its biocidal activity . One percent 8N8 produced effective bactericidal activity against Bacillus cereus, Bacillus subtilis, Haemophilus influenzae, Neisseria gonorrhoeae, Streptococcus pneumoniae, and Vibrio cholerae in 15 minutes . In contrast, most enteric gram-negative bacteria were resistant to 8N8 . One percent 8N8 was also virucidal within 15 minutes for all tested enveloped viruses, including Herpes simplex type 1, influenza A and vaccinia viruses . One percent 8N8 also demonstrated fungistatic activity on Candida albicans . The rapid and non-specific inactivation of vegetative bacteria and enveloped viruses, in addition to its fungistatic activity and low toxicity in experimental animals, makes 8N8 a potential candidate for use as a topical biocidal agent.

J Infect Dis, 2001 Jun 15, 183(12), 1819 - 21 Epub 2001 May 11.
Safety and immunogenicity of a conjugate vaccine against Haemophilus influenzae type b in splenectomized and nonsplenectomized patients with Cooley anemia; Cimaz R et al.; Patients with thalassemia are at increased risk for infections, especially after undergoing splenectomy . Vaccinations and antimicrobial prophylaxis are recommended in these patients, but the optimal immunization schedule for Haemophilus influenzae type b (Hib) vaccine is unknown . The immunogenicity of a conjugate Hib vaccine was investigated in 57 patients with thalassemia, 32 of whom had undergone splenectomy . Anti-capsular antibodies to Hib (anti-polyribosylribitol phosphate) were measured before vaccination and 2, 6, 12, 24, and 36 months after vaccination . Immunization was well tolerated . All patients achieved protective (>1 microg/mL) antibody levels . Antibody titers declined after the initial postvaccination increase, becoming undetectable in 4 patients and decreasing to concentrations of 0.15-1 microg/mL in another 2 patients when tested 2-3 years after vaccination . Hib conjugate vaccine is safe and immunogenic in patients with thalassemia major; however, additional studies are needed to assess the need and timing of booster vaccination to maintain long-term immunity.

Indian J Chest Dis Allied Sci, 2001 Jan-Mar, 43(1), 13 - 7
Drug resistant Haemophilus influenzae from respiratory tract infection in a tertiary care hospital in north India; Nag VL et al.; Haemophilus influenzae is an important respiratory pathogen . Emergence of resistance to various antibiotics is a major problem in patient management . A total of 90 strains of H . influenzae were characterized from specimens obtained from patients of acute respiratory tract infection; 13 (14.4%) belonged to type beta . On biotyping, 90% strains belonged to biotype II . The frequency of resistance to various antibiotics was as follows: cotrimoxazole 33.3% ampicillin 21.1%, cephalexin 7.8%, chloramphenicol 7.8%, ciprofloxacin 2.5% erythromycin and tetracycline 5% each . All the ampicillin-resistant strains produced beta-lactamase as detected by nitrocefin disc method . None of the strains exhibited resistance to cefaclor and third generation cephalosporins . The present study showed emergence of variable resistance to ampicillin, cotrimoxazole and other antibiotics . It is important for the clinical microbiology laboratory to monitor drug resistant strains for instituting appropriate antibiotic therapy of respiratory infections due to H . influenzae.

J Biol Chem, 2001 Aug 10, 276(32), 30315 - 25 Epub 2001 May 21.
A histidine-rich metal binding domain at the N terminus of Cu,Zn-superoxide dismutases from pathogenic bacteria: a novel strategy for metal chaperoning; Battistoni A et al.; A group of Cu,Zn-superoxide dismutases from pathogenic bacteria is characterized by histidine-rich N-terminal extensions that are in a highly exposed and mobile conformation . This feature allows these proteins to be readily purified in a single step by immobilized metal affinity chromatography . The Cu,Zn-superoxide dismutases from both Haemophilus ducreyi and Haemophilus parainfluenzae display anomalous absorption spectra in the visible region due to copper binding at the N-terminal region . Reconstitution experiments of copper-free enzymes demonstrate that, under conditions of limited copper availability, this metal ion is initially bound at the N-terminal region and subsequently transferred to an active site . Evidence is provided for intermolecular pathways of copper transfer from the N-terminal domain of an enzyme subunit to an active site located on a distinct dimeric molecule . Incubation with EDTA rapidly removes copper bound at the N terminus but is much less effective on the copper ion bound at the active site . This indicates that metal binding by the N-terminal histidines is kinetically favored, but the catalytic site binds copper with higher affinity . We suggest that the histidine-rich N-terminal region constitutes a metal binding domain involved in metal uptake under conditions of metal starvation in vivo . Particular biological importance for this domain is inferred by the observation that its presence enhances the protection offered by periplasmic Cu,Zn-superoxide dismutase toward phagocytic killing.

J Biol Chem, 2001 Aug 10, 276(32), 30326 - 34 Epub 2001 May 21.
A novel heme protein, the Cu,Zn-superoxide dismutase from Haemophilus ducreyi; Pacello F et al.; Haemophilus ducreyi, the causative agent of the genital ulcerative disease known as chancroid, is unable to synthesize heme, which it acquires from humans, its only known host . Here we provide evidence that the periplasmic Cu,Zn-superoxide dismutase from this organism is a heme-binding protein, unlike all the other known Cu,Zn-superoxide dismutases from bacterial and eukaryotic species . When the H . ducreyi enzyme was expressed in Escherichia coli cells grown in standard LB medium, it contained only limited amounts of heme covalently bound to the polypeptide but was able efficiently to bind exogenously added hemin . Resonance Raman and electronic spectra at neutral pH indicate that H . ducreyi Cu,Zn-superoxide dismutase contains a 6-coordinated low spin heme, with two histidines as the most likely axial ligands . By site-directed mutagenesis and analysis of a structural model of the enzyme, we identified as a putative axial ligand a histidine residue (His-64) that is present only in the H . ducreyi enzyme and that was located at the bottom of the dimer interface . The introduction of a histidine residue in the corresponding position of the Cu,Zn-superoxide dismutase from Haemophilus parainfluenzae was not sufficient to confer the ability to bind heme, indicating that other residues neighboring His-64 are involved in the formation of the heme-binding pocket . Our results suggest that periplasmic Cu,Zn-superoxide dismutase plays a role in heme metabolism of H . ducreyi and provide further evidence for the structural flexibility of bacterial enzymes of this class.

Arch Biochem Biophys, 2001 Jun 1, 390(1), 101 - 8
Enoyl-ACP reductase (FabI) of Haemophilus influenzae: steady-state kinetic mechanism and inhibition by triclosan and hexachlorophene; Marcinkeviciene J et al.; Steady-state kinetics, equilibrium binding, and primary substrate kinetic isotope effect studies revealed that the reduction of crotonyl-CoA by NADH, catalyzed by Haemophilus influenzae enoyl-ACP reductase (FabI), follows a rapid equilibrium random kinetic mechanism with negative interaction among the substrates . Two biphenyl inhibitors, triclosan and hexachlorophene, were studied in the context of the kinetic mechanism . IC(50) values for triclosan in the presence and absence of NAD(+) were 0.1 +/- 0.02 and 2.4 +/- 0.02 microM, respectively, confirming previous observations that the E-NAD(+) complex binds triclosan more tightly than the free enzyme . Preincubation of the enzyme with triclosan and NADH suggested that the E-NADH complex is the active triclosan binding species as well . These results were reinforced by measurement of binding kinetic transients . Intrinsic protein fluorescence changes induced by binding of 20 microM triclosan to E, E-NADH, E-NAD(+), and E-crotonyl-CoA occur at rates of 0.0124 +/- 0.001, 0.0663 +/- 0.002, 0.412 +/- 0.01, and 0.0069 +/- 0.0001 s(-1), respectively . The rate of binding decreased with increasing crotonyl-CoA concentrations in the E-crotonyl-CoA complex, and the extrapolated rate at zero concentration of crotonyl-CoA corresponded to the rate observed for the binding to the free enzyme . This suggests that triclosan and the acyl substrate share a common binding site . Hexachlorophene inhibition, on the other hand, was NAD(+)- and time-independent; and the calculated IC(50) value was 2.5 +/- 0.4 microM . Steady-state inhibition patterns did not allow the mode of inhibition to be unambiguously determined, but binding kinetics suggested that free enzyme, E-NAD(+), and E-crotonyl-CoA have similar affinity for hexachlorophene, since the k(obs)s were in the same range of 20-24 s(-1) . When the E-NADH complex was mixed with hexachlorophene ligand, concentration-independent fluorescence quenching at 480 nm was observed, suggesting at least partial competition between NADH and hexachlorophene for the same binding site . Mutual exclusivity studies, together with the above-discussed results, indicate that triclosan and hexachlorophene bind at different sites of H . influenzae FabI .

J Urban Health, 2001 Mar, 78(1), 112 - 24
Physician credentials and practices associated with childhood immunization rates: private practice pediatricians serving poor children in New York City; Hanson KL et al.; Private practice physicians in New York City's poorest neighborhoods are typically foreign trained, have generally substandard clinical practices, and have been accused of rushing Medicaid patients through to turn a profit . However, they also represent a sizable share of physician capacity in medically underserved neighborhoods . This article documents the level of credentials, systems, and immunization-related procedures among these physicians . Furthermore, it assesses the relationship between such characteristics and childhood immunization rates . The analysis utilizes a cross-sectional comparison of immunization rates in 60 private practices that submitted 2,500 or more Medicaid claims for children . Immunization data were gathered from medical records for 2,948 randomly selected children under 3 years of age . Half of sampled physicians were board certified (55%), and half were accepted by the Medicaid Preferred Physicians and Children (PPAC) program (51.7%) . Of physicians, 43% saw patients only on a walk-in basis, while only 17% scheduled the next appointment while the patient was still in the office . There were 75% of the physicians who reported usually immunizing at acute care visits . Immunization rates were higher among PPAC physicians compared to others (41% vs . 29% up to date for diphtheria and tetanus toxoids and pertussis {DTP}/Haemophilus influenzae type b {Hib}, polio, and measles-mumps-rubella {MMR}, P = .01), and board-certified physicians showed a trend toward better immunization rates (39% vs . 30%, P =.07) . Physicians who reported usually immunizing at acute care visits also had higher rates than those who did not (38% vs . 27%, P = .05) . Scheduling a date and time for the next immunization showed a trend toward association with immunization coverage (37% vs . 28%, P= .10) . Private practice physicians who provide high volumes of care reimbursed by Medicaid have improved their credentials and affiliations over time, thereby expanding reimbursement options . Credentials and affiliations were at least as effective in distinguishing relatively high- and low-performing physicians, as were immunization-related practices, suggesting that they are useful markers for higher quality care . The relative success of the PPAC program should inform efforts to improve the capacity and quality of primary care for vulnerable children . Appointment and reminder systems that effectively manage the flow of children back into the office for immunizations and the vigilant use of acute care visits for immunizations go hand in hand . Opportunity exists for payers and plans to encourage and support these actions.

Pediatr Infect Dis J, 2001 May, 20(5), 501 - 7
Bacterial and viral etiology of acute otitis media in Chilean children; Rosenblut A et al.; BACKGROUND: Acute otitis media (AOM) is a main cause for antimicrobial prescription in Latin America . Pathogen diversity in different geographic regions underscores the need for updated knowledge on AOM microbiology . AIM: To prospectively determine the role of bacteria and viruses in Chilean children with AOM . METHODS: Between July, 1998, and June, 1999, children >3 months with a presumptive diagnosis of AOM were referred to the study ear, nose and throat physician . Middle ear fluid and nasopharyngeal aspirates were obtained from children with confirmed AOM and processed for common bacteria, Mycoplasma pneumoniae, Chlamydia pneumoniae and viruses . Antimicrobial susceptibility patterns and serotypes of Streptococcus pneumoniae strains were determined . RESULTS: An ear, nose and throat physician confirmed diagnoses for 222 (42%) of 529 children referred with diagnosis of AOM, and 170 children met eligibility criteria for the study . One or more pathogens were detected in 140 of 170 (82%) children . Predominant bacteria were S . pneumoniae (37%), Haemophilus influenzae (24%) and Streptococcus pyogenes (13%) . M . catarrhalis was detected in 2 children, C . pneumoniae was found in 1 and M . pneumoniae was not detected . Viruses were detected in 22 children (13%) from nasopharyngeal aspirates, and in 6 of them the same virus was detected in middle ear fluid . Penicillin-resistant (intermediate and high) S . pneumoniae represented 40% of isolates and 10% of H . influenzae were beta-lactamase producers . All 10 penicillin-resistant S . pneumoniae strains were resistant to cefuroxime . Eighteen S . pneumoniae serotypes were detected and 19F was associated with high level penicillin resistance . CONCLUSION: This study can impact local management of AOM, and it should encourage continuous surveillance of AOM microbiology in Chile and other developing countries.

Arzneimittelforschung, 2001, 51(4), 315 - 24
Comparative in vitro activity of thiamphenicol-glycinate and thiamphenicol-glycinate-acetylcysteinate and other antimicrobials against respiratory pathogens; Drago L et al.; Thiamphenicol-glycinate-acetylcysteinate (TGA; CAS 20192-91-0) is widely used for the treatment of infections of varied aetiology . The aim of this study was to compare the antibacterial activity of thiamphenicol-glycinate (TG; CAS 15318-45-3), TGA, amoxicillin (CAS 61336-70-7) plus clavulanic acid (CAS 58001-44-8), azithromycin (CAS 83905-01-5) and ceftriaxone (CAS 104376-79-6) . Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined against Staphylococcus aureus, Klebsiella pneumoniae, Streptococcus pyogenes, Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae according to the National Committee for Clinical Laboratory Standards (NCCLS) methods . The effects of changes in assay conditions were also examined . The activity of TG and TGA was similar to that of amoxicillin plus clavulanic acid, with the exception of methicillin resistant S . aureus . Azithromycin and ceftriaxone were characterised by a limited activity against gram-positive cocci and methicillin resistant and cefinase-positive S . aureus, respectively . TG and TGA are characterized by a wide spectrum of activity, comparable to that of recent commercialized antibiotics for treatment of respiratory tract infections.

Pediatrics, 2001 Apr, 107(4), 755 - 8
Impact of the Joint Statement by the American Academy of Pediatrics/US Public Health Service on thimerosal in vaccines on hospital infant hepatitis B vaccination practices; Hurie MB et al.; OBJECTIVE: To determine the impact of the American Academy of Pediatrics/US Public Health Service (AAP/USPHS) joint statement on thimerosal in vaccines on hospital infant hepatitis B vaccination policies in Wisconsin . METHODS: The nurse managers of hospital newborn nurseries (n = 110) were surveyed by mail . Nonresponders were resurveyed . Twelve hospitals no longer provided obstetric services . Of the remaining 98 hospitals, 84 (86%) responded to the initial mailing and 14 (14%) responded to the second mailing . The number of hospitals that offered hepatitis B vaccine to infants before July 1999 was compared with that in March 2000 . The number of hospitals that had policies in place to vaccinate infants whose mothers' hepatitis B surface antigen status (HBsAg) was positive or unknown during the thimerosal alert (July 1999 through November 1999) was compared with that in March 2000 . RESULTS: Before July 1999, 81% of the hospitals representing 84% of reported Wisconsin births routinely offered hepatitis B vaccine to all infants . By March 2000, 50% of hospitals, representing 43% of births, had resumed routine infant hepatitis B vaccination . Physician decision to use a combination Haemophilus influenzae type b hepatitis B vaccine was the most frequently given reason for not reinstituting infant hepatitis B vaccination . During the thimerosal alert, 23% of hospitals did not have policies to vaccinate infants whose mothers were HBsAg-positive and 51% did not have policies to vaccinate infants whose mothers' HBsAg status was unknown . By March 2000, 6% of hospitals still did not have policies to vaccinate infants whose mothers were HBsAg-positive and 24% did not have policies to vaccinate infants whose mothers' HBsAg status was unknown . CONCLUSION: The AAP/USPHS joint statement on thimerosal in vaccines has resulted in a 38% decrease in the number of hospitals routinely offering infants hepatitis B vaccine . Although thimerosal-free hepatitis B vaccine is now available, some hospitals still do not have appropriate policies in place for vaccinating infants whose mothers' HBsAg status is positive or unknown . In the future, policymakers should include anticipated consequences that may result from changes in immunization policy in their recommendations.

Posit Health News . 1998 Spring;(No 16):18.
AquaMUNE, a brown seaweed extract, improves metabolism, immune response, energy and chelates heavy metals.
Routine immunization in HIV: helpful or harmful?
Brigham and Women's Hospital, Boston, MAAIDS: Vaccines for pneumococcus, influenza, hepatitis B virus (HBV), and Haemophilus influenzae type-B (Hib) are recommended for patients with HIV, yet new evidence indicates that some vaccinations may stimulate HIV replication . The use of vaccines has been reevaluated, however, there is little conclusive data on their efficacy and potential harm . One study of pneumococcal and influenza vaccination in HIV found the pneumococcal vaccine to be cost-effective, but the study did not consider any adverse effects the vaccine could have on HIV progression . Influenza vaccination was not found to be as cost-effective . Meanwhile, between 35 percent to 80 percent of HIV-positive patients are either immune to or are chronic carriers of HBV and are therefore not candidates for vaccination . Vaccination is recommended for those found to be HBV seronegative, even though the antibody response to the vaccine is suboptimal . Also, HIV-positive patients have a higher incidence and severity of H . influenza infection, but the efficacy of the Hib vaccine is unknown . Further studies are needed to determine the efficacy of vaccines for people with HIV and the short- and long-term effects of immunization on viral load .

Presse Med, 2001 Apr 21, 30(15), 759 - 66
{Vaccination and infection protection in patients with acquired or congenital immunodeficiency}; Lesprit P; INDICATIONS FOR VACCINATIONS IN IMMUNODEPRESSED PATIENTS: The indication for vaccination depends on the demonstration of their efficacy and tolerance . Few studies have demonstrated a clinical benefit in this heterogeneous population and generally have enrolled splenectomized patients . VACCINAL EFFICACY: Basically, the efficacy of vaccination is assessed from the post-vaccinal immune response which is generally altered in immunodepressed patients . This is specifically demonstrated for polysaccharide anti-pneumococcal vaccine . Thus, classical vaccinations with minimal immunogenic power that do not induce a memory response are not particularly effective for these patients . Indications are very controversial . PERSPECTIVES: Recently developed conjugated vaccines (anti-haemophilus, anti-pneumococcal vaccines) are more immunogenic and offer interesting perspectives for new vaccination strategies in immunodepressed patients with a major risk of severe infection or infection caused by resistant strains.

Clin Infect Dis, 2001 Jun 15, 32(12), 1700 - 5 Epub 2001 May 16.
Immunological characterization of conjugated Haemophilus influenzae type b vaccine failure in infants; Breukels MA et al.; Infant vaccination with conjugated Haemophilus influenzae type b (Hib) vaccine is highly effective in protecting against invasive Hib infections, but vaccine failures do occur . Twenty-one vaccine failures are reported since the introduction of the Hib conjugate vaccine in The Netherlands . Of the 14 evaluable patients, 6 children showed no antibody response to Hib polysaccharide in convalescent-phase serum (immunoglobulin {Ig} G anti-Hib level <1.0 microg/mL), including 1 child with hypogammaglobulinemia and 1 child with IgG2 deficiency . After revaccination, almost all children developed anti-Hib antibodies . In case of Hib vaccine failure, case investigation should be performed, including measurement of serum Ig concentrations as well as specific anti-Hib antibodies . Invasive Hib disease after infant conjugate Hib vaccination may be the presentation of an underlying immunodeficiency, but more often, only a decreased antibody response to Hib is found; revaccination with conjugated Hib vaccine is advised.

Mol Microbiol, 2001 May, 40(3), 700 - 7
Competence development by Haemophilus influenzae is regulated by the availability of nucleic acid precursors; MacFadyen LP et al.; DNA uptake by naturally competent bacteria provides cells with both genetic information and nucleotides . In Haemophilus influenzae, competence development requires both cAMP and an unidentified signal arising under starvation conditions . To investigate this signal, competence induction was examined in media supplemented with nucleic acid precursors . The addition of physiological levels of AMP and GMP reduced competence 200-fold and prevented the normal competence-induced transcription of the essential competence genes comA and rec-2 . The rich medium normally used for growth allows only limited competence . Capillary electrophoresis revealed only a subinhibitory amount of AMP and no detectable GMP, and the addition of AMP or GMP to this medium also reduced competence 20- to 100-fold . Neither a functional stringent response system nor a functional phosphoenolpyruvate:glycose phosphotransferase system (PTS) was found to be required for purine-mediated repression . Added cAMP partially restored both transcription of competence genes and competence development, suggesting that purines may reduce the response to cAMP . Potential binding sites for the PurR repressor were identified in several competence genes, suggesting that competence is part of the PUR regulon . These observations are consistent with models of competence regulation, in which depleted purine pools signal the need for nucleotides, and support the hypothesis that competence evolved primarily for nucleotide acquisition.

Kansenshogaku Zasshi, 2001 Apr, 75(4), 283 - 90
{Clinical analysis of patients with community-acquired pneumonia caused by a mixed infection of polymicrobial agents--including a comparative study of an infectious group with monomicrobial agents and an infectious group with unknown agents}; Kobashi Y et al.; We clinically analyzed 83 patients with community-acquired pneumonia caused by a mixed infection of polymicrobial agents who we have treated during the past 15 years . A comparative study among three groups; an infectious group with polymicrobial agents (83 cases), an infectious group with monomicrobial agents (335 cases), and an infectious group with unknown agents (599 cases) was performed . The results were as follows; (1) The highest percentage of patients were elderly and bedridden . (2) Striking atypical pneumonic symptoms, including dyspnea, consciousness disturbance, gastrointestinal symptoms and hypotension (shock) were present . (3) Laboratory findings of poor nutritional conditions, including decreases in serum protein, albumin, and cholineesterase, and hypoxia remarkably increased . (4) The prognosis was poor because the mortality rate (15.7%) was higher . (5) There were two polymicrobial agents for 75 patients and three agents for 8 patients . The coupling of polymicrobial agents was most frequent in five patients with Haemophilus influenzae + MSSA and five with H . influenzae + respiratory virus . These results suggest that the patients with community-acquired pneumonia caused by a mixed infection of polymicrobial agents had clinical features and causative microorganisms resembling those of elderly patients with community-acquired pneumonia . We recommended that treatment with antibiotics for them was adequate if the treatment resemble that of elderly patients.

Fam Pract, 2001 Jun, 18(3), 266 - 71
The prevalence of potential pathogenic bacteria in nasopharyngeal samples from individuals with a respiratory tract infection and a sore throat--implications for the diagnosis of pharyngotonsillitis; Gunnarsson RK et al.; BACKGROUND: Treatment failure in patients with pharyngotonsillitis after a traditional course of penicillin V is a common finding . Several factors have been proposed to explain the failure rate, but the presence of aetiological agents other than group A beta-haemolytic streptococci has attracted little attention . OBJECTIVES: The aim of the present study was to investigate if a nasopharyngeal sample could suggest the aetiology of a sore throat in patients with a respiratory tract infection . METHODS: The prevalence of potentially pathogenic bacteria (Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis) in nasopharyngeal samples from 618 healthy individuals was compared with that from 108 patients with a respiratory tract infection and a sore throat . RESULTS: The prevalence of H.influenzae was higher in patients with a sore throat than in healthy individuals of the same age . For the adult patients with a sore throat, the prevalence was 27.5% compared with 2.7% for the healthy carriers (P < 10(-7)) . The corresponding figures for schoolchildren were 31.3% versus 6.1% (P = 0.004) and for pre-school children 37.8% versus 13.2% (P = 0.0003) . CONCLUSIONS: If H.influenzae is found in a nasopharyngeal sample from a patient with a respiratory tract infection and a sore throat, it might be the aetiological agent.

Vet Microbiol, 2001 Jul 3, 81(1), 51 - 64
The transferrin receptor of Actinobacillus pleuropneumoniae: quantitation of expression and structural characterization using a peptide-specific monoclonal antibody; Bog YS et al.; When Actinobacillus pleuropneumoniae (A . pp) is grown under iron-restricted conditions in vitro, transferrin binding proteins (Tbps) are induced . The functional transferrin receptor of A . pp is composed of two outer membrane proteins (Tbp1 and Tbp2) and shows an exquisite specificity for porcine transferrin . This complex was studied using a monoclonal antibody (Mab 1.48) raised against a synthetic peptide corresponding to a hydrophilic domain of Tbp2 common to several A . pp serotypes . The antibody reacted specifically with a 60-70kDa Tbp2-antigen found in all serotypes of A . pp obtained from iron-restricted culture . It was found that Tbp2 was not expressed in iron replete medium by any serotype except serotypes 5a, 5b and 6 where a weak expression was seen . There was a weak expression of related antigens in Actinobacillus indolicus and Actinobacillus suis under iron-depleted conditions while no similar antigens were detected with the Mab in iron-starved Actinobacillus lignieresii, Actinobacillus porcinus, Actinobacillus minor, Haemophilus influenzae, and Haemophilus parasuis.Using an enzyme-linked immunosorbent assay (ELISA) based on the Mab 1.48, Tbp2 could be detected in both recombinant E . coli expressing Tbp2 and in wild type A . pp grown under iron restricted conditions . The subcellular location of Tbp2 in A . pp was studied by immunoelectron microscopy using the Mab 1.48 . Interestingly, all antibody binding was found inside the A . pp cells, while Tbp2 expressed in recombinant E . coli was found both in the cytosol and on the outer membrane . These results indicate that the Mab 1.48-reactive epitope of Tbp2 is surface exposed when it is expressed without Tbp1 in E . coli while the inaccessibility of this epitope of Tbp2 in A . pp could be due to shading by the association between Tbp2 and Tbp1.

Virchows Arch, 2001 Apr, 438(4), 362 - 9
The numbers of leukocyte subsets in lung sections differ between intercellular adhesion molecule-1-/-, lymphocyte function-associated antigen-1-/- mice and intercellular adhesion molecule-1-/- mice after aerosol exposure to Haemophilus influenzae type-b; Sinikovic B et al.; In order to investigate the role of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1) in pulmonary immunological processes, leukocyte populations were stained immunohistochemically on cryostat lung sections of ICAM-1-/- and LFA-1-/- mice . A further group of ICAM-1-/- mice was exposed to Haemophilus influenzae type-b (Hib) 24 h before being sacrificed . Comparison of the numbers of leukocytes in these groups revealed different behaviors of the leukocyte subsets: granulocytes were significantly increased in all three groups . Lymphocytes were increased in ICAM-1-/- mice, while there was no significant difference in LFA-1-/- and even a decrease in ICAM-1-/- mice after Hib exposure . Neither in ICAM-1-/- nor in LFA-1-/- mice did macrophages and dendritic cells (DCs) show significant differences to control animals . After Hib exposure, a significant elevation of DCs was observed . The following conclusions can be drawn: (1) all investigated leukocyte subsets can use ICAM-1- and LFA-1-independent pathways in the lungs of mice; (2) the pathways used by the leukocytes are cell-type specific; (3) ICAM-1 plays an important role in the enhanced recruitment of lymphocytes during Hib challenge in the lung; and (4) the alternative migratory mechanisms are able to compensate for the absence of ICAM-1 or LFA-1 or even lead to increased cell numbers . This overcompensation can be seen as a result of a balance between active alternative migratory mechanisms, which takes place in the absence of ICAM-1 or LFA-1.

Crit Care, 2001, 5(3), 167 - 73 Epub 2001 Apr 27.
Ventilator-associated pneumonia in a surgical intensive care unit: epidemiology, etiology and comparison of three bronchoscopic methods for microbiological specimen sampling; Woske HJ et al.; BACKGROUND: Ventilator-associated bacterial pneumonia (VAP) is a important intensive care unit (ICU)-acquired infection in mechanically ventilated patients . Early and correct diagnosis of VAP is difficult but is an urgent challenge for an optimal antibiotic treatment . The aim of the study was to evaluate the incidence and microbiology of ventilator-associated pneumonia and to compare three quantitative bronchoscopic methods for diagnosis . METHODS: A prospective, open, epidemiological clinical study was performed in a surgical ICU . In a prospective study, 279 patients admitted to a 14-bed surgical ICU during a 1-year period were evaluated with regard to VAP . Three quantitative culture bronchoscopic techniques for identifying the etiological agent were compared {bronchoalveolar lavage (BAL), protected specimen brush (PSB) and bronchoscopic tracheobronchial secretion (TBS)} . RESULTS: Among 103 long-term ventilated patients, 49 (48%) developed one or more VAPs (a total of 60 VAPs) . The incidence was 24 VAPs per 100 ventilated patients or 23 VAPs per 1000 ventilator days . BAL, PSB and TBS with quantitative measurements were equivalent in identifying the bacterial etiology . The VAP was caused predominantly by Staphylococcus aureus in 38% of cases, followed by Pseudomonas aeruginosa in 10%, Haemophilus influenzae in 10% and Klebsiella sp . in 9% . We did not find an increased mortality rate in patients undergoing long-term ventilation who acquired VAP in comparison with patients without VAP . CONCLUSION: For the identification of the microbiological etiology of VAP, one of three available bronchoscopic methods analysed by quantitative measurements is sufficient . In our study, quantitative bronchoscopic tracheal secretion analysis was very promising . Before accepting this method as a standard technique, other studies will have to confirm our results.

Antimicrob Agents Chemother, 2001 Jun, 45(6), 1693 - 9
Association of amino acid substitutions in penicillin-binding protein 3 with beta-lactam resistance in beta-lactamase-negative ampicillin-resistant Haemophilus influenzae; Ubukata K et al.; The affinity of {(3)H}benzylpenicillin for penicillin-binding protein (PBP) 3A was reduced in 25 clinical isolates of beta-lactamase-negative ampicillin (AMP)-resistant (BLNAR) Haemophilus influenzae for which the AMP MIC was > or =1.0 microg/ml . The affinities of PBP 3B and PBP 4 were also reduced in some strains . The sequences of the ftsI gene encoding the transpeptidase domain of PBP 3A and/or PBP 3B and of the dacB gene encoding PBP 4 were determined for these strains and compared to those of AMP-susceptible Rd strains . The BLNAR strains were classified into three groups on the basis of deduced amino acid substitutions in the ftsI gene, which is thought to be involved in septal peptidoglycan synthesis . His-517, near the conserved Lys-Thr-Gly (KTG) motif, was substituted for Arg-517 in group I strains (n = 9), and Lys-526 was substituted for Asn-526 in group II strains (n = 12) . In group III strains (n = 4), three residues (Met-377, Ser-385, and Leu-389), positioned near the conserved Ser-Ser-Asn (SSN) motif, were replaced with Ile, Thr, and Phe, respectively, in addition to the replacement with Lys-526 . The MICs of cephem antibiotics with relatively high affinities for PBP 3A and PBP 3B were higher than those of AMP and meropenem for group III strains . The MICs of beta-lactams for H . influenzae transformants into which the ftsI gene from BLNAR strains was introduced were as high as those for the donors, and PBP 3A and PBP 3B showed decreased affinities for beta-lactams . There was no clear relationship between 7-bp deletions in the dacB gene and AMP susceptibility . Even though mutations in another gene(s) may be involved in beta-lactam resistance, these data indicate that mutations in the ftsI gene are the most important for development of resistance to beta-lactams in BLNAR strains.

Antimicrob Agents Chemother, 2001 Jun, 45(6), 1688 - 92
In vivo efficacy of the new ketolide telithromycin (HMR 3647) in murine infection models; Bonnefoy A et al.; We compared the oral antibacterial activities of telithromycin (HMR 3647), a new ketolide drug, in different infections induced in mice by Staphylococcus aureus, Streptococcus pneumoniae, streptococci, enterococci, and Haemophilus influenzae with those of various macrolides and pristinamycin . Unlike all other comparators, telithromycin displayed a high therapeutic activity, particularly in septicemia induced by erythromycin A-resistant pathogens, where the ketolide was the only active compound, displaying effective doses between 3 and 26 mg/kg of body weight . Against H . influenzae, telithromycin was the most effective compound . Telithromycin displayed bacteriostatic behavior against S . pneumoniae and H . influenzae . The ketolide was also active against thigh muscle infection induced by S . aureus . The pharmacokinetic properties of telithromycin accounted for its outstanding well-balanced oral in vivo efficacy against both gram-positive cocci, whatever their phenotype of resistance, and H . influenzae.

Infect Immun, 2001 Jun, 69(6), 4180 - 4
Haemophilus ducreyi lipooligosaccharide mutant defective in expression of beta-1,4-glucosyltransferase is virulent in humans; Young RS et al.; The lipooligosaccharide (LOS) of Haemophilus ducreyi contains a major glycoform that is immunochemically identical to paragloboside, a glycosphingolipid precursor of major human blood group antigens . We recently identified the gene responsible for the glucosyltransferase activity and constructed an isogenic mutant (35000glu-) deficient in this activity . 35000glu- makes an LOS that consists only of the heptose trisaccharide core and 2-keto-deoxyoctulosonic acid (KDO) . For this study, the mutant was reconstructed in the 35000HP (human passaged {HP}) background . Five human subjects were inoculated with 35000HP and 35000HPglu- in a dose-response trial . The pustule formation rates were 40% (95% confidence interval {CI}, 13.7 to 72.6%) at 10 sites for 35000HP and 46.7% (95% CI, 24.8 to 69.9%) at 15 sites for 35000HPglu- . The histopathology and recovery rates of H . ducreyi from surface cultures and biopsies obtained from mutant and parent sites were similar . These results indicate that the expression of glycoforms with sugar moieties extending beyond the heptose trisaccharide core is not required for pustule formation by H . ducreyi in humans.

Infect Immun, 2001 Jun, 69(6), 3678 - 84
Expression of cytokine and chemokine genes by human middle ear epithelial cells induced by formalin-killed Haemophilus influenzae or its lipooligosaccharide htrB and rfaD mutants; Tong HH et al.; To define the role of nontypeable Haemophilus influenzae (NTHI) lipooligosaccharide (LOS) in the induction of proinflammatory cytokine gene expression during otitis media, we compared the abilities of formalin-killed NTHI strain 2019 and its LOS htrB and rfaD mutants to stimulate human middle ear epithelial (HMEE) cell cytokine and chemokine gene expression and production in vitro . Strain DK-1, an rfaD gene mutant, expresses a truncated LOS consisting of only three deoxy-D-manno-octulosonic acid residues, a single heptose, and lipid A . Strain B29, an isogenic htrB mutant, possesses an altered oligosaccharide core and an altered lipid A . HMEE cells were incubated with formalin-killed NTHI 2019, B29, or DK-1 . The supernatants and the cells were collected at 2, 4, 8, and 24 h after stimulation . Expression of genes for the cytokines tumor necrosis factor alpha (TNF-alpha), interleukin lbeta (IL-1beta), and IL-6 and for the chemokines macrophage inflammatory protein 1beta (MIP-1beta), monocyte chemotactic peptide 1 (MCP-1), and IL-8 was quantitated by real-time PCR . NTHI B29 did not significantly stimulate any cytokine or chemokine mRNA expression in HMEE cells . In striking contrast, NTHI 2019 induced up to 105-, 139-, and 187-fold increases in HMEE cell expression of IL-1beta, TNF-alpha, and MIP-1beta, respectively (P < 0.01 {2019 versus B29}) . NTHI 2019 also induced upregulation of IL-8, IL-6, and MCP-1 mRNA expression (by 26-, 44-, and 14-fold, respectively {P < 0.05 (2019 versus B29)}) . The significant induction of cytokine genes was confirmed by quantitating the secretion of cytokines in culture supernatants with an enzyme-linked immunosorbent assay . There were no significant differences in mRNA expression of IL-8, IL-6, and MCP-1 between the 2019- and DK-1-treated groups . The low levels of gene transcripts observed after incubation of HMEE cells with B29 indicate that products of the disrupted NTHI htrB LOS gene may play a major role in induction of these particular inflammatory mediators.

Vaccine, 2001 May 14, 19(25-26), 3600 - 5
Economic evaluation of Haemophilus influenzae type b vaccination in Slovenia; Pokorn M et al.; The objective of the present study was to assess the economical impact of invasive Haemophilus influenzae type b infections in Slovenia, where the annual incidence of these infections is 16.4/100000 in children less than 5 years of age, and to compare it with the costs of a vaccination programme . The lifetime costs and benefits were estimated for the annual birth cohort of 18200 children . In the base-case model, the calculated benefit-to-cost ratios were 0.15, 0.98 and 1.38 taking into account 95% of savings in acute care costs, medical costs, and medical and non-medical costs, respectively . From the point of view of the Institute of Health Insurance of Slovenia, who pays all healthcare and vaccination costs, the vaccination programme per annual birth cohort of 18200 children would require an extra 7023 EUR or 0.40 EUR per cohort-child . The savings to society would represent 118410 EUR, indicating the rationale for inclusion of H . influenzae type b vaccination in the routine childhood immunisation programme in Slovenia.

Vaccine, 2001 May 14, 19(25-26), 3399 - 407
Formulation and characterisation of Bordetella pertussis fimbriae as novel carrier proteins for Hib conjugate vaccines; Crowley-Luke A et al.; Haemophilus influenzae type b (Hib) capsular polysaccharide (polyribosylribitol phosphate, PRP) is the active component of conjugate vaccines that have proven successful in preventing invasive Hib disease . Conjugation of PRP to a protein carrier greatly improves its immunogenicity providing protection in infants and subsequent antibody maturation upon boosting . In this study, fimbriae isolated from Bordetella pertussis have been assessed as novel carrier proteins . These proteins are components of some acellular pertussis vaccines and clinical trials have indicated that fimbriae could be important protective antigens against whooping cough . Fimbriae (Fim2 and Fim3) purified from B . pertussis were dissociated in 6 M guanidine hydrochloride, pH 10.5, to produce proteins of defined size and to facilitate the production and characterisation of the conjugates . Both carbodiimide-mediated coupling and reductive amination were used to conjugate PRP to dissociated fimbriae . Efficiency of conjugation was determined by size exclusion chromatography followed by protein and polysaccharide analysis of fractionated components . Immunisation of rabbits with dissociated fimbriae-PRP conjugates (D.fim-PRP) produced high anti-fimbrial and anti-PRP IgG titres . Use of a D.fim-PRP conjugate could protect against Hib disease and may also augment protection against B . pertussis.

New Microbiol, 2001 Apr, 24(2), 117 - 24
Morphological and biochemical variations of Haemophilus influenzae type b induced by pH and temperature changes; Oliva B et al.; Haemophilus influenzae type b ATCC 10211 was cultured at different temperatures (25 degrees C-49 degrees C) and pH values (5.7-8.7) either in liquid or semisolid medium . Morphological variations of individual cells were noted by optical microscopy depending upon the conditions of growth . At higher temperatures filaments were produced whereby the length of individual cells increased compared to cultures grown at 37 degrees C . Filaments were also observed at lower pH values . Culture conditions also affected colonial morphology . At low pH values colonies had an enhanced lobulated contour and were more wrinkly and rougher than at higher pH . The changes in cellular and colonial morphology were correlated with distinct outer membrane protein profiles . The changes in temperature and pH did not affect identification of the microorganism by the API system.

Scand J Infect Dis, 2001, 33(4), 266 - 71
Prevalence of and detection of resistance to ampicillin and other beta-lactam antibiotics in Haemophilus influenzae in Denmark; Arendrup M et al.; The susceptibility of Haemophilus influenzae to penicillin V and G, ampicillin and cefuroxime was investigated by MIC, disc and tablet diffusion methods, using chocolate agar as test medium, to determine the prevalence of ampicillin-resistant isolates and the optimal method for their detection . Eighty-six isolates were clinical isolates collected prospectively from July to September 1998 and 22 isolates were clinical isolates with decreased susceptibility to ampicillin previously referred to the reference laboratory . Eighty-seven isolates were ampicillin-susceptible and 16 were ampicillin-resistant . Thirteen produced beta-lactamases . Among the consecutive isolates 12.8% were resistant . With each of the Rosco Neo-sensitabs containing penicillin G, 2.5 microg ampicillin and 33 microg ampicillin, 3 very major errors occurred (resistant isolates misinterpreted as susceptible) and 5-13 major errors (susceptible isolates misinterpreted as resistant) . The AB biodisk containing ampicillin (10 microg) was superior to the penicillin V and G discs, i.e . only 1 very major error occurred and major and minor errors were infrequent . The cefuroxime disc identified 4/8 beta-lactamase-negative ampicillin-resistant isolates . Thus, for susceptibility testing with chocolate agar as test medium, the use of an inoculum of 10(5) colony-forming units, 10 microg ampicillin discs and interpretative zone diameters of > or = 28 mm indicating susceptibility and < or = 25 mm indicating resistance was found to produce reliable identification of ampicillin-resistant isolates of H . influenzae.

Scand J Infect Dis, 2001, 33(4), 263 - 5
Haemophilus influenzae osteomyelitis in adults: a report of 4 frontal bone infections and a review of the literature; Sarria JC et al.; Haemophilus influenzae occasionally causes hematogenous long-bone osteomyelitis in children . In adults, however, bone infections caused by this organism are extremely rare . We report four adult cases of H . influenzae frontal bone osteomyelitis and review 12 cases from the literature.

Protein Sci, 2001 Mar, 10(3), 592 - 8
Solution structure of the DNA-binding domain of the TyrR protein of Haemophilus influenzae; Wang Y et al.; The TyrR protein of Haemophilus influenzae is a 36-kD transcription factor whose major function is to control the expression of genes important in the biosynthesis and transport of aromatic amino acids . Using (1)H and (15)N NMR spectroscopy, we have determined the 3D solution structure of the TyrR C-terminal DNA-binding domain (DBD) containing residues from 258 to 318 (TyrR{258-318}) . The NMR results show that this segment of TyrR consists of a potential hinge helix at its N terminus (residues 263-270) as well as three well-defined alpha-helices extending from residues 277-289 (HR-2), 293-300 (HR-1), and 304-314 (HR) . Helix HR-1 and HR fold in a typical helix-turn-helix (HTH) motif . The three helices and the hinge helix are tightly bound together by hydrophobic interaction and hydrogen bonds . Several hydrophilic residues whose side chains may directly interact with DNA are identified . A hydrophobic patch that may be part of the interaction surface between the domains of TyrR protein is also observed . Comparisons with the structures of other HTH DNA-binding proteins reveal that in terms of the spatial orientation of the three helices, this protein most closely resembles the cap family.

J Med Microbiol, 2001 May, 50(5), 472 - 5
A comparison of the performance of bacitracin-incorporated chocolate blood agar with chocolate blood agar plus a bacitracin disk in the isolation of Haemophilus influenzae from sputum; Nye KJ et al.; The lack of selectivity of chocolated blood agar (CBA), routinely used for the isolation of Haemophilus influenzae, may lead to masking of the growth of H . influenzae due to overgrowth of competing flora . Bacitracin can be used as a selective agent, either incorporated into the medium or applied to the medium in a filter paper . However, neither method has been evaluated or compared in a large study . Sputum samples (1990) were examined in four laboratories and the isolation rates of H . influenzae on chocolated blood agar with bacitracin added to the medium (BCA) and chocolated blood agar (CBA) with a bacitracin disk were compared . A plain blood agar plate was also inoculated to facilitate the isolation of Streptococcus pneumoniae so that its effects on the isolation of H . influenzae could be assessed . No significant difference was found between the isolation rates of H . influenzae on BCA and CBA with a bacitracin disk, although competing flora was greatly reduced and quantity of growth of H . influenzae increased on BCA . The presence of S . pneumoniae did not affect the isolation of H . influenzae in this study.

J Med Microbiol, 2001 May, 50(5), 449 - 55
The effect of temperature on the interaction of Haemophilus ducreyi with human epithelial cells; Makakole SC et al.; To investigate if temperature affects the interaction of Haemophilus ducreyi with human epithelial cells, nine strains were used to evaluate the adhesion kinetics of the organism at 33 degrees C and 37 degrees C . The effect of the free toxin on the epithelial cells at those temperatures was also assessed . The cyto-adherence kinetics of H . ducreyi to the epithelial cells was significantly greater at 33 degrees C (10 times more) than at 37 degrees C in all seven clinical isolates tested . There was a significant difference in cell-associated H . ducreyi at 33 degrees C as compared with 37 degrees C . Control strains showed similar adhesion properties at both temperatures . However, the virulent strain CIP542 adhered in larger amounts than the avirulent strain A77 . Electron microscopy revealed that there was more tissue necrosis at the lower than the higher temperature . The effect of the free toxin was the same at each temperature . However, strain A77 had significantly lower toxicity than strain CIP542 and the clinical isolates . These results suggest that H . ducreyi displays a temperature-dependent interaction with human epithelial cells, and this feature may play a role in the virulence of the organism in vivo . While the overall toxic effect of viable bacteria depends on the metabolic activity of the bacteria and is, therefore, higher at 33 degrees C than at 37 degrees C withthe same initial inoculum, the effect of the extracted toxin at molecular level with fixed concentrations is a temperature-independent event.

Manag Care Interface, 2001 Apr, 14(4), 68 - 80
Evidence-based guidelines for treatment of bacterial respiratory tract infections in the era of antibiotic resistance; Jacobs MR et al.; Antimicrobial resistance in bacterial respiratory tract pathogens is a rapidly evolving and increasingly disconcerting problem . Major factors that have contributed to resistance are inappropriate prescribing of antibiotics for viral infections and the use of antibiotics with poor activity . The treatment of respiratory tract infections is significantly affected by resistance in organisms such as Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis . Resistance to beta-lactams, sulfonamides, and macrolides continues to rise . Evidence-based guidelines, founded on clinical and bacteriological outcomes, are imperative to treat patients effectively, to limit the spread of these pathogens, and to minimize further development of resistance . Pharmacokinetic and pharmacodynamic parameters have recently been shown to correlate with clinical outcome, and offer a more rational approach to predicting antimicrobial efficacy and determining clinically relevant susceptibility breakpoints.

Int J Antimicrob Agents, 2001 May, 17(5), 401 - 5
Effect of different antibacterial agents and surfactant protein-A (SP-A) on adherence of some respiratory pathogens to bronchial epithelial cells; Ferrara A et al.; Some antibiotics at sub-inhibitory concentrations are able to alter bacterial surface structures and modulate adhesiveness by affecting the expression of microbial adhesins . An important mechanism of pulmonary defence against pathogens is SP-A, one of the proteins of the alveolar surfactant having opsonizing activity . The aim of this study was to investigate the effect that sub-inhibitory concentrations of different antibiotics and physiological concentrations of SP-A (1 and 5 microg/ml) could exert on the adherence of respiratory pathogens to the bronchial epithelial cell line, WI26VA4 . Cefdinir and clarithromycin showed high efficacy, mainly at 1/2 MIC, in reducing the adherence of Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae strains to values lower or equal to 50% of the control; sparfloxacin showed the same effect on S . aureus and S . pneumoniae but teicoplanin only on S . pneumoniae . Other similar results were observed with netilmicin on Klebsiella pneumoniae (40%) and with cefepime and ciprofloxacin on Pseudomonas aeruginosa (60%) . Clarithromycin reduced the adherence of K . pneumoniae to 80% although it is not active against this strain . Adherence of the test strains was not modified by SP-A alone or in combination with any of the antibiotics used.

Int J Antimicrob Agents, 2001 May, 17(5), 365 - 70
Evaluation of two in vitro pharmacodynamic simulation models: microfiltration versus centrifugation-filtration; Alou L et al.; Pharmacodynamic in vitro models that simulate serum antimicrobial concentrations provide more information about the activity of an antibiotic than MICs or traditional time-kill methods . The aim of this study was to compare two pharmacodynamic simulation models using ATCC strains of five different species and five antibiotics . In the first model (Centriprep-10 system), a filtration-centrifugation process was used to eliminate the antibiotic; in the second model (microfiltration system) no centrifugation was necessary . The antibiotic concentrations tested were similar to those in serum after normal doses of cefuroxime, clarithromycin, ciprofloxacin, gentamicin and cefotaxime . No significant differences were observed in the killing rates between the models except in the case of Haemophilus influenzae and cefotaxime . The new microfiltration model had the following advantages: lack of the carry-over effect, the absence of centrifugation that could damage bacteria and the possibility of increasing the number of incubation periods to give a better fit of the kinetic profile of man.

Trends Microbiol, 2001 May, 9(5), 193 - 6
SecB, a molecular chaperone with two faces; Driessen AJ; SecB is a molecular chaperone unique to the phylum Proteobacteria, which includes the majority of known Gram-negative bacteria of medical, industrial and agricultural significance . SecB is involved in the translocation of secretory proteins across the cytoplasmic membrane . The crystal structure of the Haemophilus influenzae SecB provides new insights into how SecB simultaneously recognizes its two ligands: unfolded preproteins and SecA, the ATPase subunit of the translocase . SecB uses its entire molecular surface for these two functions, but for preprotein release and its own membrane release, SecB relies on the catalytic activity of SecA . This defines SecB as a translocation-specific molecular chaperone.

J Autoimmun, 2001 May, 16(3), 257 - 62
The mimicry of human glycolipids and glycosphingolipids by the lipooligosaccharides of pathogenic neisseria and haemophilus; Harvey HA et al.; It has been known for many years that bacteria can induce autoimmune responses in humans resulting in serious disease . Recent work has shown that a number of bacteria that colonize human mucosal surfaces exclusively express antigens on their surfaces which are molecular mimics of glycosphingolipids found on human cells . These structures are important in the pathogenesis of Neisseria and Haemophilus species for both immune evasion and in the adherence and invasion of human cells . There is no evidence that colonization or infections by these bacterial species is associated with autoimmune disease .

J Autoimmun, 2001 May, 16(3), 219 - 27
Multiple pathways to induction of virus-induced autoimmune demyelination: lessons from Theiler's virus infection; Miller SD et al.; Infection of SJL mice with wild-type BeAn strain of Theiler's murine encephalomyelitis virus (TMEV) leads to CD4(+)T cell-mediated CNS demyelination characterized by the development of anti-myelin epitope autoimmune responses via epitope spreading during the chronic stage of disease . To exmine the feasibility of virus-encoded mimic epitopes to initiate CNS autoimmunity, we recently developed a molecular mimicry model of virus-induced demyelinating disease wherein a non-pathogenic variant strain of TMEV was engineered to encode a 30-mer peptide encompassing the immunodominant myelin proteolipid protein, PLP139-151, epitope . SJL mice infected intracerebrally with TMEV encoding either the native PLP139-151 determinant or various peptide mimics of the epitope develop an early onset demyelinating disease mediated by activated PLP139-151-specific Th1 cells . The autoimmune nature of this early-onset demyelinating disease is shown by the fact that induction of tolerance to the PLP139-151 peptide prevents clinical disease and associated PLP139-151-specific T cell responses without affecting T cell reactivity to virus epitopes . Most significantly, TMEV encoding a molecular mimic peptide derived from the Haemophilus influenzae bacteria, homologous at only six out of thirteen of the core amino acids, led to CNS disease . These studies provide conclusive evidence that virus-induced myelin-specific autoreactive T cells can be induced by molecular mimicry and provide a useful model to study the disease inducing ability of viruses encoding human-disease-related mimicry peptides .

Am J Prev Med, 2001 May, 20(4 Suppl), 75 - 83
Undervaccination with hepatitis B vaccine: missed opportunities or choice?
Jiles RB, Daniels D, Yusuf HR, McCauley MM, Chu SY.
BACKGROUND: An estimated 1 million to 1.25 million people in the United States are chronically infected with hepatitis B virus (HBV) and are at substantially increased risk of developing chronic liver disease, including cirrhosis and primary hepatocellular carcinoma . Immunization with hepatitis B vaccine (HepB) is the most effective means of preventing HBV infection and its consequences . METHODS: To identify and describe children who had not completed the three-dose HepB series, we analyzed data from the 1999 National Immunization Survey (NIS) . Among the 2648 children aged 19 to 35 months who did not complete the HepB series, we examined the relationship between the number of doses of HepB received and the number of vaccination visits made, receipt of the birth dose of HepB, age at the time of first vaccination visit (excluding that for the birth dose of HepB), and completion of the 4:3:1:3 series (four doses of diphtheria and tetanus toxoids and pertussis vaccine, three doses of poliovirus vaccine, one dose of measles-containing vaccine, and three doses of Haemophilus influenzae type b vaccine {Hib}) . RESULTS: Overall, 11.8% of the children who were included in the 1999 NIS did not complete the HepB series . Among these series-incomplete children, most (79.8%; 95% CI, 77.4%-82.2%) did not receive the birth dose of HepB, and most (80.2%; 95% CI, 77.6%-82.8%) had three or more vaccination visits . Most of the series-incomplete children (87.3%; 95% CI, 85.1%-89.5%) who had three or more vaccination visits received one or two doses of HepB . Among series-incomplete children with at least three vaccination visits, those who did not receive any HepB were more likely to have completed the 4:3:1:3 series (67.1%; 95% CI, 58.8%-75.4%) than those who received at least one dose of HepB (52.7%; 95% CI, 49.0%-56.4%) . CONCLUSIONS: Children who did not complete the HepB series fell into three distinct groups: children who made at least three vaccination visits but did not begin the HepB series (n=326); children who made three or more vaccination visits and received one or two doses of HepB (n=1835); and children who made fewer than three vaccination visits (n=487).Different intervention strategies are needed to have an impact on each of these groups, including understanding why parents and providers may not be receptive to HepB, decreasing missed opportunities to administer HepB, and implementing tracking systems such as registries to identify and contact children who are due or overdue for vaccinations.

Am J Prev Med, 2001 May, 20(4 Suppl), 61 - 8
Undervaccinated African-American preschoolers: a case of missed opportunities; Daniels D et al.; OBJECTIVE: To identify factors associated with undervaccination of African-American preschoolers, to describe the number of vaccination visits made by undervaccinated children and the number of visits needed to be series complete, and to describe the children who did not receive the single dose of measles-containing vaccine recommended for preschoolers . METHODS: We used the 1999 National Immunization Survey (NIS) to describe vaccination coverage for the 4:3:1:3 vaccine series (four doses of diphtheria and tetanus toxoids and pertussis vaccine, three doses of poliovirus vaccine, one dose of any measles-containing vaccine, and three doses of Haemophilus influenzae type b vaccine) among non-Hispanic, African-American preschoolers due to concerns that they may be at risk of undervaccination . Children who did not complete this basic vaccine series were classified for further analysis according to the number of doses they lacked (i.e., one dose missed, two or three doses missed, or four or more doses missed) . Significant associations between demographic characteristics and vaccination status or degree of undervaccination were determined . RESULTS: Of the 26.2% of African-American preschoolers who did not complete the 4:3:1:3 vaccine series, 40.3% lacked one, 35.3% lacked two or three, and 25.0% lacked four or more doses of vaccine . Children who did not complete the 4:3:1:3 vaccine series were less likely to have married mothers, were less likely to have mothers aged > or = 35 years, or were less likely to be up to date at age 3 months than the children who completed the 4:3:1:3 vaccine series . Among the undervaccinated, 63.7% had a sufficient number of vaccination visits to have completed the basic series . However, most (78.7%) of the severely undervaccinated (children who lacked more than three doses of vaccine) had three or fewer vaccination visits . For 72.6% of the undervaccinated preschoolers, only one additional vaccination visit was needed to complete the 4:3:1:3 vaccine series; among these, 78.3% had an adequate number of vaccination visits to have completed the series . Overall, 9.9% of the African-American children aged 19 to 35 months (i.e., approximately 85,000 African-American children aged 19 to 35 months) were at risk for measles . Among the children who lacked more than three doses of vaccine, 68.1% were at risk . CONCLUSIONS: Our study suggests that the estimated coverage of 73.8% for the 4:3:1:3 vaccine series among African-American children aged 19 to 35 months was not a result of limited access to care . On the contrary, 90.5% of African-American children had enough vaccination visits to complete the series . To raise coverage and prevent potential outbreaks, providers should assess each child's vaccination status at every visit, and administer all needed vaccinations at that time . For the most severely undervaccinated children, this strategy may not be adequate, because they did not have the minimum number of vaccination visits required for series completion . For these children, other strategies are needed for increasing vaccination coverage.

Am J Prev Med, 2001 May, 20(4 Suppl), 47 - 54
Vaccination status of children in the Women, Infants, and Children (WIC) Program: are we doing enough to improve coverage?
Shefer AM, Luman ET, Lyons BH, Coronado VG, Smith PJ, Stevenson JM, Rodewald LE.
BACKGROUND: Vaccination-promoting strategies in the Supplemental Nutrition Program for Women, Infants, and Children (WIC) have been shown to produce dramatic improvements in coverage and other health outcomes . OBJECTIVES: To determine national and state-specific population-based vaccine coverage rates among preschool children who participate in the WIC program, and to describe the strategies for promoting vaccination in WIC . DESIGN/METHODS: Demographic data, WIC participation, and vaccination histories for children aged 24 to 35 months in 1999 were collected from parents through the National Immunization Survey . The healthcare providers for the children in the survey were contacted to verify and complete vaccination information . We defined children as up-to-date (UTD) if they had received four doses of diphtheria and tetanus toxoids and pertussis vaccine (DPT), three doses of poliovirus vaccine, one dose of measles-mumps-rubella vaccine (MMR), and three doses of Haemophilus influenzae type b vaccine (Hib) by 24 months . Description of state-level vaccination-promoting activities in WIC was collected through an annual survey completed by the state WIC and immunization program directors . RESULTS:Complete data were collected on 15,766 children, of whom 7783 (49%) participated in WIC sometime in their lives . Nationally, children who had ever participated in WIC were less well-immunized at 24 months compared to children who had not: 72.9% UTD (95% CI, 71.3-74.5) versus 80.8% UTD (95% CI, 79.5-82.1), respectively . In 42 states, 24-month coverage among WIC participants was less than among non-WIC participants, including 13 states where the difference was > or = 10% . Vaccination activities linked with WIC were reported from 76% of 8287 WIC sites nationwide . States conducting more-frequent interventions and reaching a higher proportion of WIC participants had 40% higher vaccination coverage levels for the WIC participants in that state (p<0.05) . CONCLUSIONS: Children served by WIC remain less well-immunized than the nation's more-affluent children who do not participate in WIC . Thus, WIC remains a good place to target these children . This study provides evidence that fully implemented WIC linkage works to improve vaccination rates . Strategies that have been shown to improve the vaccination coverage levels of WIC participants should be expanded and adequately funded to protect these children.

Am J Prev Med, 2001 May, 20(4 Suppl), 28 - 31
Changes in vaccination coverage estimates among children aged 19-35 months in the United States, 1996-1999; Barker LE et al.; BACKGROUND: Childhood vaccinations have a major impact on the reduction and elimination of many causes of morbidity and mortality among children . Monitoring of annual vaccination coverage levels over time is necessary to characterize undervaccination . Here, coverage estimates for 1996 (1997 for varicella) were compared with those of 1999 . METHODS: Immunization coverage among children aged 19 to 35 months in 1996 (1997 for varicella) and 1999 for a variety of vaccines and vaccine series were compared using Wald chi-square tests and data from the National Immunization Survey . RESULTS: Record high immunization coverage among children aged 19 to 35 months in the United States has increased by a statistically significant amount between 1996 and 1999 for diphtheria, tetanus, and pertussis; measles, mumps, and rubella; Haemophilus influenzae type b; hepatitis B; and standard series made up of these individual vaccines . Coverage with the vaccine for varicella dramatically increased between 1997 and 1999 . However, between 1996 and 1999, coverage with three or more doses of polio vaccine decreased by a small but statistically significant amount . CONCLUSION: Despite the drop for polio vaccine, coverage remains high . Continued monitoring is required to determine if the drop in polio coverage is a cause for concern.

J Antimicrob Chemother, 2001 May, 47(5), 681 - 4
Comparative in vitro activity of the new quinolone gemifloxacin (SB-265805) with other fluoroquinolones against respiratory tract pathogens; Garcia-Garrote F et al.; The in vitro activity of gemifloxacin (SB-265805) was compared with that of other fluoroquinolones against 302 clinical isolates of Streptococcus pneumoniae, 300 clinical isolates of Haemophilus influenzae and 28 clinical isolates of Moraxella catarrhalis, including multiply resistant strains . Gemifloxacin at 0.12 mg/L inhibited all microorganisms tested . MIC(90) values of gemifloxacin, trovafloxacin, grepafloxacin and levofloxacin against all (630) isolates tested were 0.03, 0.12, 0.12 and 1 mg/L, respectively . MIC(90) values of the same fluoroquinolones against S . pneumoniae were 0.06, 0.25, 0.12 and 1 mg/L, respectively.

J Antimicrob Chemother, 2001 May, 47(5), 675 - 80
In vitro activity of linezolid and 11 other antimicrobials against 566 clinical isolates and comparison between NCCLS microdilution and Etest methods; Tubau F et al.; The in vitro activity of linezolid and 11 other antimicrobials was determined for 566 clinical isolates of Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus spp., Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, some of them resistant to several antibiotics, using a broth microdilution method and the Etest method . All Gram-positive organisms tested were inhibited by a concentration of <or=4 mg/L of linezolid, including methicillin-resistant staphylococci, vancomycin- and ampicillin-resistant enterococci, and penicillin-intermediate and -resistant pneumococci . MICs of linezolid by the Etest method were usually one to two dilution values lower than those obtained by microdilution.

J Antimicrob Chemother, 2001 May, 47(5), 605 - 10
Carriage of antibiotic-resistant bacteria by healthy children; Millar MR et al.; The frequency of carriage of antibiotic-resistant bacteria in healthy 7- and 8-year-old children in Bristol was studied . Children born in Avon between 1 April 1991 and 31 December 1992, attending the Avon Longitudinal Study of Pregnancy and Childhood (ALSPAC) 7 year follow-up clinic, formed the study population . Carriage was estimated using mouth and stool samples . None of 105 children on whom information was available had received tetracycline, chloramphenicol, ciprofloxacin or an extended-spectrum cephalosporin in the previous year . Staphylococcus aureus was isolated from mouthwashes from 200 (37.1%) of 539 children sampled . Six (3%) of the isolates were resistant to chloramphenicol or tetracycline and four (2%) were methicillin resistant . Haemophilus spp . were isolated from 369 (72%) of 513 samples and 63 (17%) were ampicillin resistant, 49 (13.3%) were erythromycin resistant and seven (1.9%) were tetracycline resistant . Branhamella catarrhalis was isolated from 333 (74%) of 450 samples . Twenty-eight (8.4%) were erythromycin resistant and 14 (4.2%) strains were tetracycline resistant . Group A beta-haemolytic streptococci were isolated from 17 of 507 children sampled . One (5.9%) was tetracycline resistant . Stool samples were returned from 335 (62%) of 539 children from whom they were requested . Eleven per cent of samples yielded Gram-negative bacilli with high-level resistance to chloramphenicol, which was frequently linked to resistance to ampicillin, spectinomycin and streptomycin . Isolates demonstrating resistance to the third-generation cephalosporin ceftazidime were recovered from 17 subjects (3.2%) . Six (35%) of 17 isolates possessed extended-spectrum beta-lactamases . Healthy children carry bacteria resistant to antibiotics to which children are not usually exposed . Resistance to ceftazidime, chloramphenicol and tetracycline may be co-selected by exposure to other antibiotics used in children or may be acquired from family members, pets, other children or food . These results suggest that antibiotic-resistant bacteria are widely disseminated and may be acquired by children before exposure to specific selection pressure.

J Antimicrob Chemother, 2001 May, 47(5), 565 - 73
Antibacterial activity of essential oils and their major constituents against respiratory tract pathogens by gaseous contact; Inouye S et al.; The antibacterial activity of 14 essential oils and their major constituents in the gaseous state was evaluated against Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus pyogenes and Staphylococcus aureus . For most essential oils examined, H . influenzae was most susceptible, followed by S . pneumoniae and S . pyogenes, and then S . aureus . Penicillin-susceptible and -resistant S . pneumoniae were comparable in susceptibility . Escherichia coli, which was used as a control, showed least susceptibility . A minimal inhibitory dose (MID) was introduced as a measure of the vapour activity . Among 14 essential oils, cinnamon bark, lemon-grass and thyme oils showed the lowest MID, followed by essential oils containing terpene alcohols as major constituents . The essential oils containing terpene ketone, ether and, in particular, hydrocarbon had high MIDS.The vapour activity on short exposure was comparable to that following overnight exposure, and rapid evaporation was more effective than slow evaporation of essential oils . The vapour concentration and absorption into agar of essential oils reached a maximum 1 or 2 h after rapid evaporation . These results indicate that the antibacterial action of essential oils was most effective when at high vapour concentration for a short time.

J Clin Microbiol, 2001 May, 39(5), 1941 - 6
New tests for syphilis: rational design of a PCR method for detection of Treponema pallidum in clinical specimens using unique regions of the DNA polymerase I gene; Liu H et al.; A sensitive and specific PCR method to detect Treponema pallidum in clinical specimens was developed . PCR primers were designed based on two unique features of the DNA polymerase I gene (polA) . The first distinctive characteristic is that the region codes for a high cysteine content and has low homology with similar regions of DNA polymerase I gene from known microorganisms . The second unique feature is the presence of four insertions in the gene . PCR tests using primers designed on the basis these regions reacted with various pathogenic T . pallidum subspecies but did not react with nonpathogenic treponemal species or other spirochetes . An additional 59 species of bacteria and viruses, including those that cause genital ulcers, tested negative . This PCR method is extremely robust and sensitive . The detection limit is about 10 to 25 organisms when analyzed on gel . However, the analytic sensitivity can be increased by at least 1 log, to a detection limit of a single organism, when the ABI 310 Prism Genetic Analyzer is used to detect fluorescence-labeled amplicons . We further used this test in a clinical setting and compared the results with results from a previously reported multiplex-PCR test (for T . pallidum, Haemophilus ducreyi, and herpes simplex virus) . We tested 112 genital ulcer specimens by the polA PCR, obtaining a sensitivity of 95.8% and a specificity of 95.7% . These results suggest that the polA PCR is applicable as a routine clinical diagnostic test for syphilis.

J Clin Microbiol, 2001 May, 39(5), 1757 - 62
Outer membrane proteins and DNA profiles in strains of Haemophilus parasuis recovered from systemic and respiratory sites; Ruiz A et al.; Polyserositis caused by Haemophilus parasuis is an important disease that affects mostly weaned pigs . Recent studies have shown that virulence can differ among strains recovered from distinct body sites and also that it may be related to the presence of certain outer membrane proteins (OMPs) . The objective of this study was to compare the OMP and DNA profiles of H . parasuis strains isolated from systemic and respiratory sites from diseased and healthy pigs . Strains evaluated in this study were processed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and repetitive-PCR techniques . Two experiments were conducted in order to better define the relationship among genotype, phenotype, and site of isolation . Experiment 1 included 53 H . parasuis isolates recovered from healthy and diseased pigs from unrelated herds . Experiment 2 included 31 isolates of H . parasuis obtained from diseased pigs involved in an outbreak in a large, multifarm system . Results showed that strains recovered from systemic sites had more homogeneous OMP and DNA profiles than those isolated from respiratory sites . Evaluation of isolates involved in the multifarm outbreak showed that only two H . parasuis strains were causing disease . These strains had homogeneous OMP and DNA profiles . However, it was noted that these two parameters were unrelated, since strains classified in the same genotype group expressed different OMP profiles . The homogeneity of OMP and DNA profiles of strains isolated from systemic sites strongly suggests the existence of clonal relationships between virulent strains and also suggests that expression of certain OMP profiles may be related to virulence.

Drugs, 2001, 61(4), 443 - 98
Review of macrolides and ketolides: focus on respiratory tract infections; Zhanel GG et al.; The first macrolide, erythromycin A, demonstrated broad-spectrum antimicrobial activity and was used primarily for respiratory and skin and soft tissue infections . Newer 14-, 15- and 16-membered ring macrolides such as clarithromycin and the azalide, azithromycin, have been developed to address the limitations of erythromycin . The main structural component of the macrolides is a large lactone ring that varies in size from 12 to 16 atoms . A new group of 14-membered macrolides known as the ketolides have recently been developed which have a 3-keto in place of the L-cladinose moiety . Macrolides reversibly bind to the 23S rRNA and thus, inhibit protein synthesis by blocking elongation . The ketolides have also been reported to bind to 23S rRNA and their mechanism of action is similar to that of macrolides . Macrolide resistance mechanisms include target site alteration, alteration in antibiotic transport and modification of the antibiotic . The macrolides and ketolides exhibit good activity against gram-positive aerobes and some gram-negative aerobes . Ketolides have excellent activity versus macrolide-resistant Streptococcus spp . Including mefA and ermB producing Streptococcus pneumoniae . The newer macrolides, such as azithromycin and clarithromycin, and the ketolides exhibit greater activity against Haemophilus influenzae than erythromycin . The bioavailability of macrolides ranges from 25 to 85%, with corresponding serum concentrations ranging from 0.4 to 12 mg/L and area under the concentration-time curves from 3 to 115 mg/L x h . Half-lives range from short for erythromycin to medium for clarithromycin, roxithromycin and ketolides, to very long for dirithromycin and azithromycin . All of these agents display large volumes of distribution with excellent uptake into respiratory tissues and fluids relative to serum . The majority of the agents are hepatically metabolised and excretion in the urine is limited, with the exception of clarithromycin . Clinical trials involving the macrolides are available for various respiratory infections . In general, macrolides are the preferred treatment for community-acquired pneumonia and alternative treatment for other respiratory infections . These agents are frequently used in patients with penicillin allergies . The macrolides are well-tolerated agents . Macrolides are divided into 3 groups for likely occurrence of drug-drug interactions: group 1 (e.g . erythromycin) are frequently involved, group 2 (e.g . clarithromycin, roxithromycin) are less commonly involved, whereas drug interactions have not been described for group 3 (e.g . azithromycin, dirithromycin) . Few pharmacoeconomic studies involving macrolides are presently available . The ketolides are being developed in an attempt to address the increasingly prevalent problems of macrolide-resistant and multiresistant organisms.

Kansenshogaku Zasshi, 2001 Mar, 75(3), 193 - 200
{Clinical analysis of patients with community-acquired pneumonia requiring hospitalization classified by age group}; Kobashi Y et al.; We classified 1017 patients with community-acquired pneumonia requiring hospitalization experienced in Kawasaki Medical School Kawasaki Hospital during the past 15 years into five age groups (< or = 54 years old, 55-64 years old, 65-74 years old, 75-84 years old, > or = 85 years old) . With particular emphasis on the elderly patients, we then compared the clinical and microbiological findings in the five groups . The results were as follows; (1) Half of patients in the over 85 years old group were bed-ridden . (2) The proportion receiving antibiotics before hospitalization decreased with age . (3) There were striking atypical pneumonic symptoms, such as dyspnea and consciousness disturbance in the two age groups over 75 years old . (4) Hypotension (shock) increased with age . (5) Markers of nutritional conditions, such as serum protein, albumin, cholinesterase, and hypoxia remarkably increased in the two age groups over 75 years old . (6) There were no significant differences in the isolation rate of etiological microorganisms . (7) The number of polymicrobial agents in the < or = 54 years old group was lower than that in the other age groups . (8) Mycoplasma pneumoniae was most significantly higher in < or = 54 years old group, Haemophilus influenzae in patients 55-64 years old, and Streptococcus pneumoniae in both 65-74 and 75-84 years old groups . (9) The isolation rate of MSSA, gram-negative bacilli such as Klebsiella pneumoniae, Pseudomonas aeruginosa, respiratory viruses increased with age . (10) The amount of sepsis increased with age . (11) The prognosis was poor in the two groups over 75 years old because the mortality rate (over 10%) was higher that for the other age groups.

Clin Infect Dis, 2001 May 15, 32 Suppl 2, S81 - 93
Worldwide prevalence of antimicrobial resistance in Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis in the SENTRY Antimicrobial Surveillance Program, 1997-1999; Hoban DJ et al.; The in vitro activities of numerous antimicrobials against clinical isolates of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis from patients with bloodstream and respiratory tract infections in the United States, Canada, Europe, Latin America, and the Asia-Pacific region were studied in the SENTRY Antimicrobial Surveillance Program . Penicillin resistance (minimum inhibitory concentration, > or =2 microg/mL) was noted in all 5 geographic regions, and a high and increasing rate of macrolide resistance among S . pneumoniae isolates was observed . Elevated rates of resistance to clindamycin, trimethoprim-sulfamethoxazole, chloramphenicol, and tetracycline were seen . beta-Lactamase-mediated resistance in H . influenzae to amoxicillin and variable trimethoprim-sulfamethoxazole resistance by region were documented . Resistance to several drugs continues to emerge among pneumococci worldwide, but more stable resistance patterns have been noted for H . influenzae and M . catarrhalis . Continued surveillance of this pathogen group appears to be prudent.

Ann Dermatol Venereol, 2001 Mar, 128(3 Pt 2), 326 - 33
{Diagnostic criteria for erysipelas}; Vaillant L; Diagnosis of erysipelas is based upon the association of an acute inflammatory plaque with fever, lymphagiitis, adenopathy and hyperleukocytosis . These associated symptoms are variable (20-70 p . 100 of cases) . Bacteriology is not helpful for the diagnosis of erysipelas because of a low sensitivity (hemoculture 5 p . 100, standard examinations 5-41 p . 100), or delayed positivity (serology) . Moreover cutaneous bacteriology is difficult to assess when other bacteria than streptococci are isolated . Erysipelas have to be distinguished from non-necrotizing cellulitis by peculiar clinical features (such as erysipeloid, facial staphylococcal infection, Pasteurella, Haemophilus influenzae) and from necrotizing fasciitis . Some non-infectious diseases may mimic erysipelas such as venous thrombosis, familial Mediterranean fever, prosthesis intolerance, and compartment syndrome . Because the diagnostic value of clinical symptoms is not known and no diagnostic gold standard has been established, it is impossible to be sure that non-streptococcal erysipelas (especially staphylococcal) really exists . Thus, the first line treatment for all erysipelas must be an antistreptococcal antibiotic . Before prescribing a treatment, hemoculture and blood cell count could be useful . If antistreptococcal antibiotherapy is inefficient, all the differential diagnoses must be reviewed.

Clin Microbiol Infect, 2001 Mar, 7(3), 114 - 9
Effect of some fractions of alveolar surfactant (phospholipids and SP-A) on the bactericidal activity of different antimicrobials against some respiratory pathogens; Ferrara A et al.; OBJECTIVES: To investigate the effects of physiologic concentrations, at alveolar level, of some fractions of pulmonary surfactant (phospholipids and SP-A) on the bactericidal activity of different antimicrobials against some respiratory pathogens . METHODS: The antimicrobial agents cefdinir, sparfloxacin, clarithromycin, teicoplanin, cefepime, ciprofloxacin, netilmicin and tobramycin, depending on their specific activity, were investigated against Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Klebsiella pneumoniae and Pseudomonas aeruginosa . Killing curves were carried out with antimicrobials at 0.5 and 2 MIC, SP-A at 1 and 5 mg/L and phospholipids at 50 mg/L . RESULTS: Time-kill experiments showed that while SP-A never modified the activity of antimicrobials, phospholipids exerted, in some cases, a weak antagonistic effect . Among antibacterials and pathogens investigated, phospholipids were able to decrease the rate of killing of cefepime and ciprofloxacin only on P . aeruginosa, both at 0.5 and at 2 MIC, with an increase of about 1 log in CFU . The combination of SP-A and phospholipids never modified the effect observed in the presence of lipids alone . CONCLUSIONS: The paucity of data only allow us to observe that the examined antibiotics do not have substantially reduced activity against respiratory pathogens studied in the presence of physiologic concentrations of some fractions of surfactant . Cefepime alone already exerted a small effect, and ciprofloxacin at 2 MIC, even in the presence of phospholipids, retained its bactericidal activity.

Clin Microbiol Infect, 2001 Mar, 7(3), 107 - 13
Vertebral infections caused by Haemophilus aphrophilus: case report and review; Colson P et al.; OBJECTIVE: To review in detail clinical presentation, bacteriologic findings, associated conditions and treatment of Haemophilus aphrophilus vertebral osteomyelitis and to compare them to a case we report herein . METHODS: A Medline (National Library of Medicine) search of the literature was performed by using the key words H . aphrophilus, spondylodiscitis, discitis, and vertebral osteomyelitis . The references of the case reports were examined for additional cases, especially those cited in older articles that had not been entered onto the bibliographic database . RESULTS: A case report of spondylodiscitis due to H . aphrophilus in a 35-year-old patient with a history of dental abscess 7 months before admission is presented . The patient responded well to treatment with ceftriaxone and ciprofloxacin . To date, only 14 cases of H . aphrophilus vertebral osteomyelitis have been reported . They are usually reported in middle-aged patients, usually male . Most recent cases have been treated with fluoroquinolones . Duration of treatment usually ranges from 1 to 3 months . CONCLUSIONS: H . aphrophilus is an uncommon cause of vertebral osteomyelitis . Patients are regularly cured by antibiotic therapy, provided that a tissue biopsy is performed in order to isolate the causative bacterium.

AIDS, 2001 Mar 30, 15(5), 635 - 9
Vitamin A and risk of HIV-1 seroconversion among Kenyan men with genital ulcers; MacDonald KS et al.; BACKGROUND: Vitamin A is involved in normal immune function and the maintenance of mucosal integrity through complex effects on cellular differentiation . OBJECTIVE: We sought to determine whether serum vitamin A levels were associated with altered susceptibility to primary infection with HIV-1 in men with high-risk sexual behaviour and genital ulcers who presented for treatment at an STD clinic in Nairobi, Kenya . METHODS: HIV-1 seronegative men were prospectively followed . Vitamin A levels at study entry were compared among 38 men who HIV-1 seroconverted versus 94 controls who remained HIV seronegative . RESULTS: Vitamin A deficiency (retinol less than 20 microg/dl) was very common and was present in 50% of HIV-1 seroconverters versus 76% of persistent seronegatives . Seroconversion was independently associated with a retinol level greater than 20 microg/dl (HR 2.43, 95% CI 1.25-4.70, P = 0.009), and a genital ulcer aetiology caused by Haemophilus ducreyi (HR 3.49, 95% CI 1.03-11.67, P = 0.04) . Circumcision was independently associated with protection (HR 0.46, 95% CI 0.23-0.93, P = 0.03) . CONCLUSION: Vitamin A deficiency was not associated with an increased risk of HIV-1 infection among men with concurrent STD . A decreased risk of HIV-1 seroconversion was independently associated with lower retinol levels . The effects of vitamin A on macrophage and lymphoid cell differentiation may paradoxically increase mucosal susceptibility to HIV-1 in some vulnerable individuals, such as men with genital ulcers . Lack of circumcision and chancroid are confirmed as important co-factors for heterosexual HIV-1 transmission . The role of vitamin A in heterosexual HIV-1 transmission requires further study.

Biometrics, 1999 Dec, 55(4), 1306 - 13
A hierarchical Bayesian model to predict the duration of immunity to Haemophilus influenzae type b; Auranen K et al.; A hierarchical Bayesian regression model is fitted to longitudinal data on Haemophilus influenzae type b (Hib) serum antibodies . To estimate the decline rate of the antibody concentration, the model accommodates the possibility of unobserved subclinical infections with Hib bacteria that cause increasing concentrations during the study period . The computations rely on Markov chain Monte Carlo simulation of the joint posterior distribution of the model parameters . The model is used to predict the duration of immunity to subclinical Hib infection and to a serious invasive Hib disease.

Otol Neurotol, 2001 Jan, 22(1), 11 - 4
Goblet cell density in acute otitis media caused by Moraxella catarrhalis; Caye-Thomasen P et al.; HYPOTHESIS AND BACKGROUND: Secretory otitis media is associated with a highly increased goblet cell density, confirming the secretory pathogenesis of this disease . Previous studies have shown that the middle ear goblet cell density, and thus the secretory capacity, are massively increased during experimental acute otitis media and at least 6 months thereafter, conceivably predisposing to the subsequent development of secretory otitis media . These studies used middle ear inoculation of either Streptococcus pneumoniae, nontypeable Haemophilus influenzae, or H . influenzae type b . The present study aimed at determining the goblet cell density during and after acute otitis media caused by Moraxella catarrhalis to clarify whether this bacterium induces an equivalently enhanced secretory capacity . METHODS: Twenty-five 25 rat middle ears were inoculated with M . catarrhalis . Five rats were killed on days 4, 8, 16, 60, and 180 after inoculation, followed by staining, dissection, and whole-mount embedding of the middle ear mucosae . The goblet cell density was determined by counting in 24 fields, covering the entire middle ear . RESULTS: In comparison with 25 normal middle ears, the goblet cell density was significantly increased in almost all counting localities, from day 4 and < or = 2 months after inoculation . The goblet cell density peaked on day 16, subsided thereafter, and in some areas reached a normal level 6 months after the acute incident . Mucosal areas containing goblet cells were consistently enlarged, thus leaving the middle ear with an increased secretory capacity during and 6 months after inoculation . CONCLUSION: The goblet cell density of the middle ear mucosa is increased during acute otitis media caused by M . catarrhalis and up to several months thereafter . This may predispose to the subsequent development of secretory otitis media . However, in comparison with acute otitis media caused by other bacteria, M . catarrhalis induced only modest changes in goblet cell density.

Vaccine, 2001 Apr 30, 19(23-24), 3189 - 200
Effect of physico-chemical modification on the immunogenicity of Haemophilus influenzae type b oligosaccharide-CRM(197) conjugate vaccines; Bolgiano B et al.; Haemophilus influenzae type b (Hib) poly-ribosyl-ribityl phosphate (PRP) oligosaccharide-CRM(197) conjugate vaccines from two different manufacturers (Hib A and Hib B) were subjected to adverse storage conditions and used to establish correlates between physico-chemical characteristics and immunogenicity . There were manufacturer-specific differences in the effect of freezing or freeze-thawing on the carrier protein conformation and the anti-CRM(197) or anti-PRP IgG response in rabbits whereas both conjugates showed similar stability when stored at elevated temperatures . Both oligosaccharide-CRM(197) conjugate vaccines formed apparent 'aggregates' of non-specifically associated higher molecular weight material when subjected to elevated temperatures or repeated freeze-thawing . Following subcutaneous injection of samples into CBA mice and New Zealand White rabbits, the amount of IgG raised against CRM(197) was significantly lower for samples incubated at 37 or 55 degrees C compared with those kept at 4 degrees C, consistent with the less well-folded conformation of the carrier protein observed at elevated temperatures . Moreover, there was a parallel reduction in the amount of IgG raised against PRP and the level of bactericidal antibodies induced by vaccines A and B stored at 55 degrees C consistent with the observed depolymerisation of the oligosaccharide chains . Carrier protein conformational changes resulting from storage under adverse conditions did not affect the immunogenicity to Hib PRP in laboratory animals unless associated with loss of bound saccharide presumably because the carrier protein retains continuous T(H) cell epitopes which are unaffected by conformational changes.

Vaccine, 2001 Apr 30, 19(23-24), 3058 - 66
The adjuvant effect of synthetic oligodeoxynucleotide containing CpG motif converts the anti-Haemophilus influenzae type b glycoconjugates into efficient anti-polysaccharide and anti-carrier polyvalent vaccines; von Hunolstein C et al.; Synthetic oligodeoxynucleotides containing CpG immunostimulatory sequences (ISS) have been shown to act as potent adjuvants of type 1 immune responses when co-administered with protein or peptide vaccines . We have recently shown that ISS can increase the anti-polysaccharide (CHO) and anti-tetanus toxoid (TT) or anti-diphtheria (CRM) toxoid antibody levels if used as adjuvant of anti-Haemophilus influenzae type b (Hib) CHO vaccine conjugated with TT or CRM . The analysis of anti-TT and anti-CRM IgG subclasses showed a significant increase in IgG2a, IgG2b and/or IgG3 in the presence of ISS . Anti-TT and anti-CRM antibodies were shown to neutralize the activity of both the tetanus and diphtheria toxin in vivo or in vitro tests respectively . These data show that ISS have the potential to increase host antibody response against both the CHO and the protein component of a conjugated vaccine, and encourage the investigation to identify strategies of vaccination with schedules aimed at the valuation of protein carriers as protective immunogens.

Microb Drug Resist, 2001 Spring, 7(1), 33 - 8
A multicenter study of the antimicrobial susceptibility of Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis isolated from patients with community-acquired lower respiratory tract infections in 1999 in Portugal; Melo-Cristino J et al.; A nationwide multicenter study (including 25 laboratories) of the antimicrobial susceptibility of bacterial pathogens commonly associated with community-acquired lower respiratory tract infections (LRTI), with testing undertaken in a central laboratory, was conducted in Portugal in 1999 . Antimicrobial resistance in Haemophilus influenzae has not increased in the last decade . Of the 498 isolates tested, 12.4% produced beta-lactamase and >95% were susceptible to all antimicrobials except ampicillin . In contrast, there was a rapid increase of resistance in Streptococcus pneumoniae . Of the 312 isolates tested, 24.7% exhibited decreased susceptibility to penicillin (13.5% showed low-level and 11.2% high-level resistance), 13.8% were resistant to erythromycin, clarithromycin and azithromycin, and 13.6% to cefuroxime and to tetracycline . Of the 38 Moraxella catarrhalis tested, 81.6% produced beta-lactamase . Resistance to penicillin, cefuroxime, erythromycin, clarithromycin, and azithromycin in S . pneumoniae and beta-lactamase production in H . influenzae were significantly higher in pediatric patients than in adults . Overall, amoxycillin/clavulanate was the most active antimicrobial agent in vitro against H . influenzae, S . pneumoniae, and M . catarrhalis isolated from patients with community-acquired LRTI in Portugal.

Medicine (Baltimore), 2001 Mar, 80(2), 75 - 87
Community-acquired pneumonia . A prospective outpatient study; Bochud PY et al.; We initiated a prospective study with a group of practitioners to assess the etiology, clinical presentation, and outcome of community-acquired pneumonia in patients diagnosed in the outpatient setting . All patients with signs and symptoms suggestive of pneumonia and an infiltrate on chest X-ray underwent an extensive standard workup and were followed over 4 weeks . Over a 4-year period, 184 patients were eligible, of whom 170 (age range, 15-96 yr; median, 43 yr) were included and analyzed . In 78 (46%), no etiologic agent could be demonstrated . In the remaining 92 patients, 107 etiologic agents were implicated: 43 were due to "pyogenic" bacteria (39 Streptococcus pneumoniae, 3 Haemophilus spp., 1 Streptococcus spp.), 39 were due to "atypical" bacteria (24 Mycoplasma pneumoniae, 9 Chlamydia pneumoniae, 4 Coxiella burnetii, 2 Legionella spp.), and 25 were due to viruses (20 influenza viruses and 5 other respiratory viruses) . There were only a few statistically significant clinical differences between the different etiologic categories (higher age and comorbidities in viral or in episodes of undetermined etiology, higher neutrophil counts in "pyogenic" episodes, more frequent bilateral and interstitial infiltrates in viral episodes) . There were 2 deaths, both in patients with advanced age (83 and 86 years old), and several comorbidities . Only 14 patients (8.2%) required hospitalization . In 6 patients (3.4%), the pneumonia episode uncovered a local neoplasia . This study shows that most cases of community-acquired pneumonia have a favorable outcome and can be successfully managed in an outpatient setting . Moreover, in the absence of rapid and reliable clinical or laboratory tests to establish a definite etiologic diagnosis at presentation, the spectrum of the etiologic agents suggest that initial antibiotic therapy should cover both S . pneumoniae and atypical bacteria, as well as possible influenza viruses during the epidemic season.

Genome Biol . 2001;2(4):RESEARCH0013 . Epub 2001 Mar 22.
Conservation of the binding site for the arginine repressor in all bacterial lineages; Makarova KS et al.; BACKGROUND: The arginine repressor ArgR/AhrC is a transcription factor universally conserved in bacterial genomes . Its recognition signal (the ARG box), a weak palindrome, is also conserved between genomes, despite a very low degree of similarity between individual sites within a genome . Thus, the arginine repressor is different from two other universal transcription factors - HrcA, whose recognition signal is very strongly conserved both within and between genomes, and LexA/DinR, whose signal is strongly conserved within, but not between, genomes . The arginine regulon is well studied in Escherichia coli and to some extent in Bacillus subtilis and some other genomes . Here, we apply the comparative genomic approach to the prediction of the ArgR-binding sites in all completely sequenced bacterial genomes . RESULTS: Orthologs of ArgR/AhrC were identified in the complete genomes of E . coli, Haemophilus influenzae, Vibrio choleras, B . subtilis, Mycobacterium tuberculosis, Thermotoga maritima, Chlamydia pneumoniae and Deinococcus radiodurans . Candidate arginine repressor binding sites were identified upstream of arginine transport and metabolism genes . CONCLUSIONS: We found that the ArgR/AhrC recognition signal is conserved in all genomes that contain genes encoding orthologous transcription factors of this family . All genomes studied except M . tuberculosis contain ABC transport cassettes (related to the Art system of E . coli) belonging to the candidate arginine regulons.

Biochemistry, 2001 Apr 24, 40(16), 5041 - 8
Reduction precedes cytidylyl transfer without substrate channeling in distinct active sites of the bifunctional CDP-ribitol synthase from Haemophilus influenzae; Zolli M et al.; CDP-ribitol synthase is a bifunctional reductase and cytidylyltransferase that catalyzes the transformation of D-ribulose 5-phosphate, NADPH, and CTP to CDP-ribitol, a repeating unit present in the virulence-associated polysaccharide capsules of Haemophilus influenzae types a and b {Follens, A., et al . (1999) J . Bacteriol . 181, 2001} . In the work described here, we investigated the order of the reactions catalyzed by CDP-ribitol synthase and conducted experiments to resolve the question of substrate channeling in this bifunctional enzyme . It was determined that the synthase first catalyzed the reduction of D-ribulose 5-phosphate followed by cytidylyl transfer to D-ribitol 5-phosphate . Steady state kinetic measurements revealed a 650-fold kinetic preference for cytidylyl transfer to D-ribitol 5-phosphate over D-ribulose 5-phosphate . Rapid mixing studies indicated quick reduction of D-ribulose 5-phosphate with a lag in the cytidylyl transfer reaction, consistent with a requirement for the accumulation of K(m) quantities of D-ribitol 5-phosphate . Signature motifs in the C-terminal and N-terminal sequences of the enzyme (short chain dehydrogenase/reductase and nucleotidyltransferase motifs, respectively) were targeted with site-directed mutagenesis to generate variants that were impaired for only one of the two activities (K386A and R18A impaired for reduction and cytidylyl transfer, respectively) . Release and free diffusion of the metabolic intermediate D-ribitol 5-phosphate was indicated by the finding that equimolar mixtures of K386A and R18A variants were efficient for bifunctional catalysis . Taken together, these findings suggest that bifunctional turnover occurs in distinct active sites of CDP-ribitol synthase with reduction of D-ribulose 5-phosphate and release and free diffusion of the metabolic intermediate D-ribitol 5-phosphate followed by cytidylyl transfer.

Nippon Rinsho, 2001 Apr, 59(4), 688 - 93
{Beta-lactamase negative ampicillin-resistant Haemophilus influenzae(BLNAR)}; Yamamoto K et al.; The affinity of {3H}-benzylpenicillin for penicillin-binding protein(PBP) 3A/3B was reduced in clinical isolates beta-lactamase-negative ampicillin(ABPC)-resistant Haemophilus influenzae(BLNAR) with MIC of > or = 1 microgram/ml to ABPC . The sequence of ftsI gene encoding the transpeptidase domain of PBP3A/3B were determined for these strains, and compared to those of ABPC-susceptible Rd strain . Common substitutions of deduced amino acid residues were identified in transpeptidase region on the ftsI gene in BLNAR strains . Homology modeling of the three-dimensional structure of PBP3 showed that every common substitution occurred at active site pocket surrounded by three conserved motifs . The MICs of beta-lactams for H . influenzae transformants in which ftsI gene from BLNAR was introduced, were as high as those for the donors and PBP3A/3B showed a decreased affinity for beta-lactams . These data indicate that mutations in the ftsI gene are the most important for development of resistance to beta-lactams in BLNAR.

Pediatr Infect Dis J, 2001 Mar, 20(3), 356 - 61
Cefepime in the empiric treatment of meningitis in children; Saez-Llorens X et al.; BACKGROUND: Because the introduction of extended spectrum cephalosporins into pediatric practice offers a number of choices for treatment, we review efficacy studies of cefepime monotherapy in the treatment of bacterial meningitis in children . METHODS: Two open, randomized, comparative studies assessed the efficacy of cefepime empiric monotherapy in the treatment of bacterial meningitis in 345 pediatric patients . These studies were conducted in Latin America and compared cefepime (50 mg/kg/dose every 8 h) with either cefotaxime (50 mg/kg/dose every 6 h) or ceftriaxone (50 mg/kg/dose every 12 h) . Patients 2 months to 14 years old who had clinical signs and symptoms consistent with a central nervous system infection were enrolled . Efficacy was based on clinical and bacteriologic response . RESULTS: Integrated results from the Latin American studies indicated a 75% cure rate with cefepime vs . a 78% cure rate with comparator, among evaluable patients . Overall the rate of treatment failure was 12% . Haemophilus influenzae had the highest bacterial eradication rate (97% overall), and rates were comparable in cefepime and comparator arms . Eradication rates for Neisseria meningitidis were equally high in both treatment arms (95% overall), and the eradication rate for Streptococcus pneumoniae was 92% overall . Of the patients with S . pneumoniae isolated during pretreatment (from either cerebrospinal fluid or blood), 11 (16 isolates in total) had their isolates tested against penicillin and all were susceptible . Presence or absence of seizures, level of consciousness, Glasgow Coma Score and duration of signs and symptoms were strong predictors of outcome . Collectively no specific safety concerns were identified . CONCLUSION: Cefepime represents an important therapeutic option for the empiric treatment of bacterial meningitis in children, based on the good clinical response and bacteriologic eradication rates observed in this review.

Pediatr Infect Dis J, 2001 Mar, 20(3), 300 - 5
Non-type b Haemophilus influenzae disease: clinical and epidemiologic characteristics in the Haemophilus influenzae type b vaccine era; Heath PT et al.; BACKGROUND: As a result of the decline in Haemophilus influenzae type b (Hib) disease caused by the widespread use of conjugate vaccines, non-type b H . influenzae will become a more important cause of H . influenzae (Hi) disease . Characterization of the clinical and epidemiologic features of non-b Hi disease is needed in the Hib vaccine era . METHODS: A prospective active surveillance study of invasive Hi disease involving pediatricians in the United Kingdom and Republic of Ireland . For the first phase of the study (October 1, 1992, to October 31, 1995) pediatricians were asked to report any child who had invasive Hi disease and who had received Hib conjugate vaccine . For the second phase of the study (November 1, 1995 . To December 31, 1998) pediatricians were asked to report any child with invasive Hi disease regardless of vaccination status . RESULTS: During the study period 102 cases of invasive non-type b Hi disease and 106 cases of invasive Hib disease were reported in children who had been fully vaccinated against Hib . Children with non-type b disease were younger (16 vs . 22 months of age, P = 0.08), less likely to have meningitis and epiglottitis (P < or = 0.001) and more likely to have pneumonia and bacteremia (P < or = 0.001) than children with type b disease . For the last 2 years of the study invasive Hi disease occurring in a fully vaccinated child was more likely to be caused by a non-b strain than by a type b strain (58 vs . 38) . In 1998 the incidence of non type-b Hi disease in all children <5 years of age in the UK was 1.3/100,000 as compared with an incidence of Hib disease of 0.6/100,000 . The majority (88%) of non-b strains isolated in children were nontypable strains . CONCLUSIONS: Non-b Hi is a rare cause of disease in children, but in the Hib vaccine era it has become more common than type b as a cause of Hi disease in fully vaccinated children.

Pediatr Infect Dis J, 2001 Mar, 20(3), 260 - 4
Bacteriologic and clinical efficacy of trimethoprim-sulfamethoxazole for treatment of acute otitis media; Leiberman A et al.; BACKGROUND: Trimethoprim-sulfamethoxazole (T/S) has often been used as first and second line of treatment for acute otitis media (AOM) . Because of the increasing resistance of Streptococcus pneumoniae and Haemophilus influenzae to T/S, we undertook the present study to investigate the bacteriologic and clinical efficacy of this drug in AOM . METHODS: Fifty-four culture-positive evaluable patients ages 3 to 32 months with AOM were treated with T/S 4/20 mg/kg in two divided daily doses for 10 days . Middle ear fluid (MEF) was cultured at enrollment (Day 1) and on Days 4 and 5 after initiation of treatment . Additional MEF cultures were obtained if clinical relapse occurred . Clinical failure was determined when the symptoms and signs of AOM did not improve or recurred during therapy . Bacteriologic failure was defined by positive culture on Days 4 and 5, or negative on Days 4 and 5 but positive again before the end of treatment . Patients were followed until Day 28 +/- 2 . RESULTS: A total of 67 organisms were isolated from MEF specimens of the 54 study patients: S . pneumoniae, 24; H . influenzae, 40; and Streptococcus pyogenes, 3 . Fifteen (63%) of 24 S . pneumoniae were nonsusceptible to T/S (trimethoprim MIC, >0.5 microg/ml), of which 10 (67%) were highly resistant to T/S (trimethoprim MIC, > or = 4.0 microg/ml) . Twelve (30%) of 40 H . influenzae and all 3 S . pyogenes isolates were nonsusceptible to T/S (MIC > or = 4.0 microg/ml) . Bacteriologic eradication occurred in 9 of 9 (100%) and 27 of 27 (100%) T/S-susceptible S . pneumoniae and H . influenzae, respectively, vs . 4 of 15 (27%) and 6 of 12 (50%) T/S-nonsusceptible S . pneumoniae and H . influenzae, respectively (P < 0.001) . The 3 patients with S . pyogenes failed bacteriologically . Nine new organisms, not initially isolated, emerged during treatment, 7 of which (77%) were resistant to T/S . Altogether bacteriologic failure (organisms not eradicated plus newly emerged) occurred in 29 (53%) of 54 patients . Clinical failures occurred in 8 (15%) of 54 patients, and in 7 of these 8 cases the clinical failures occurred in those with bacteriologic failures . Ten patients relapsed clinically after completion of treatment and in 8 of them tympanocentesis for MEF culture was performed . Six of these 8 cultures were positive, and the initial pathogen was isolated in 4 of 6 (67%) . CONCLUSIONS: A high bacteriologic failure rate as well as a considerable clinical failure rate occurred among patients with AOM treated with T/S . We believe that T/S is no longer an appropriate empiric choice for the treatment of AOM in regions where high T/S resistance among respiratory pathogens is reported.

Pediatr Infect Dis J, 2001 Mar, 20(3), 247 - 50
Antibiotic-resistant bacteria in pediatric chronic sinusitis; Slack CL et al.; BACKGROUND: Limited information exists on emerging bacterial resistance patterns in pediatric chronic sinusitis . METHODS: A retrospective review (1995 to 1998) of the aerobic microbiology of chronic sinusitis in children at a tertiary care children's hospital was conducted . One hundred nineteen children (mean age, 4.9 years) with maxillary sinusitis of >8 weeks duration and no known immunodeficiency or cystic fibrosis who underwent antral irrigation were included . RESULTS: One hundred sixty-one of 240 (67%) aerobic cultures were positive, yielding 274 isolates . Eighty-eight positive cultures were polymicrobial . The most frequent isolates were nontypable Haemophilus influenzae (24%), Streptococcus pneumoniae (19%), Moraxella catarrhalis (17%), coagulase-negative Staphylococcus (6%), alpha-streptococci (6%), diphtheroids (5%), Staphylococcus aureus (3%) and Neisseria spp . (3%) . Rates of nonsusceptibility of Streptococcus pneumoniae were 64% for penicillin (24% high grade resistance), 40% for cefotaxime, 18% for clindamycin and 0% for vancomycin . Rates of nonsusceptibility of S . pneumoniae did not change significantly during the study period . Thirty-nine percent of H . influenzae isolates were beta-lactamase-positive and 44% were nonsusceptible to ampicillin (41% high grade resistance) . Beta-lactamase positivity of H . influenzae decreased during the study period (P = 0.06) . All M . catarrhalis isolates tested were beta-lactamase-positive . CONCLUSION: This study indicates that the aerobic pathogens in pediatric chronic sinusitis include bacteria typical of acute sinusitis as well as organisms more characteristic of chronic disease . Moreover it highlights the significant role of antibiotic-resistant aerobes, including multiply resistant S . pneumoniae, in pediatric chronic sinusitis.

Pediatr Infect Dis J, 2001 Mar, 20(3), 235 - 9
Imperfect memory and the development of Haemophilus influenzae type B disease; Lucas AH et al.; Considerable evidence indicates that both anticapsular antibody and immunologic memory play a role in immunity to Haemophilus influenzae type b (Hib) disease . The efficacy of memory (or antibody) cannot be expected to be 100%; therefore some individuals may develop invasive disease despite their having been naturally primed . The proportion of cases of H . influenzae type b disease with evidence of immunologic memory is related to both the efficacy of memory in preventing disease and the age-related prevalence of memory in the population . The task is to discern the relative contributions of antibody and memory in conferring protection and to determine the extent to which natural exposure and vaccination establish these two effector mechanisms.

Antimicrob Agents Chemother, 2001 May, 45(5), 1585 - 8
Identification of beta-lactamase-negative, ampicillin-resistant strains of Haemophilus influenzae with four methods and eight media; Barry AL et al.; A challenge set of 143 non-beta-lactamase-producing strains of Haemophilus influenzae was tested for ampicillin susceptibility on two broth media and six agar media, using broth microdilution, agar dilution, disk diffusion, and E-test procedures . When beta-lactamase-negative, ampicillin-resistant (BLNAR) strains were defined as those for which the ampicillin MIC was > or = 4.0 microg/ml, 5 to 44% of our selected strains were BLNAR depending on the medium and/or test method used . If nonsusceptible strains for which ampicillin MICs were intermediate were included in the BLNAR category, 32 to 50% of our isolates would be considered BLNAR . These data emphasize the need for a standardized testing procedure and a universal definition of BLNAR strains before the clinical relevance of such strains can be evaluated . NCCLS dilution tests with haemophilus test medium broth or agar are preferred for testing ampicillin against H . influenzae.

Antimicrob Agents Chemother, 2001 May, 45(5), 1463 - 6
Activities of BMS 284756 (T-3811) against Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae isolates from SENTRY antimicrobial surveillance program medical centers in Latin America (1999); Gales A et al.; The antimicrobial activity of BMS 284756, a novel des-F(6)-quinolone, was comparatively evaluated against 257 Streptococcus pneumoniae, 198 Haemophilus influenzae, and 88 Moraxella catarrhalis strains isolated in Latin America between July and September of 1999 as part of the SENTRY Antimicrobial Surveillance Program . Nearly 28.0% of S . pneumoniae strains were nonsusceptible to penicillin . The rank order of quinolone potency versus S . pneumoniae was BMS 284756 (MIC at which 90% of isolates were inhibited {MIC(90)}, 0.12 microg/ml) > trovafloxacin (MIC(90), 0.25 microg/ml) > gatifloxacin (MIC(90), 0.5 microg/ml) > levofloxacin and ciprofloxacin (MIC(90), 1 to 2 microg/ml) . All S . pneumoniae strains that were not susceptible to other quinolones were inhibited by BMS 284756 at < or = 2 microg/ml . The overall prevalence of beta-lactamase production was 15.2% in H . influenzae and 98.9% in M . catarrhalis . BMS 284756 showed excellent potency and spectrum against this group of pathogens, inhibiting all isolates at < or = 0.12 microg/ml . BMS 284756 exhibited activity similar to those displayed by the new fluoroquinolones, such as levofloxacin, trovafloxacin, or gatifloxacin, and could be a therapeutic option for empirical treatment of community-acquired respiratory tract infections.

Biol Blood Marrow Transplant, 2001, 7(3), 171 - 83
Vaccination against infectious disease following hematopoietic stem cell transplantation; Avigan D et al.; Patients undergoing hematopoietic stem cell transplantation (HSCT) experience a prolonged period of dysfunctional immunity associated with an increased risk of bacterial and viral infections . Effective approaches toward vaccinating patients against common pathogens are being explored but are limited by poor levels of responsiveness . Relevant studies examining the nature of reconstitution of cellular and humoral immunity and its impact on vaccination strategies against infectious pathogens are reviewed . Following transplantation, deficiencies in cellular immunity are characterized by the inversion of CD4/CD8 ratios, a decreased proliferative response to mitogens, and the development of anergy to recall antigens as measured by delayed-type hypersensitivity testing . The impact on humoral immunity consists of decreased levels of circulating immunoglobulin, impaired immunoglobulin class switching, and a loss of complexity in immunoglobulin gene rearrangement patterns . In this setting, a loss of protective immunity has been demonstrated against viral and bacterial pathogens previously targeted by childhood vaccination . Infections due to encapsulated bacterial organisms such as Streptococcus pneumoniae and Haemophilus influenzae type B remain prevalent even in the late posttransplantation period . The efficacy of vaccination following HSCT is influenced by the time elapsed since transplantation, the nature of the hematopoietic graft, the use of serial immunization, and the presence of graft-versus-host disease . Strategies to enhance vaccine efficacy include pretransplantation immunization of the stem cell donor and the use of cytokine adjuvants.

Infect Dis Clin North Am, 2001 Mar, 15(1), 9 - 19
Standards for immunization practice for vaccines in children and adults; Peter G et al.; Administration of vaccines is a continuing challenge . In childhood immunizations, many of the goals for national coverage rates by 2000 were achieved and the goal of annual influenza immunization for adults 65 years of age and older was reached . These successes in childhood immunization rates have led to record low numbers of cases of many vaccine-preventable diseases, such as measles and Haemophilus influenzae, type b invasive disease . These diseases will recur, however, as evidenced by the measles epidemic of 1989-1991, if high immunization coverage is not maintained . The development of immunization delivery systems to sustain these high rates in young children is essential to ensure that the 11,000 infants born each day in the United States receive all recommended vaccines, as noted in the recent NVAC report on strategies to sustain success in childhood immunization . For adults, the total economic burden of treating these vaccine-preventable diseases is estimated to exceed $10 billion each year, reflecting in part widespread underuse of vaccines in adults and resulting missed opportunities to prevent diseases such as influenza and pneumococcal infection . The development of standards for immunization practices in children and adults has been an important component in meeting these challenges and ensuring appropriate delivery of vaccines . Periodic review and updating is necessary and revision of the standards for adults by the NCAI and NVAC, pediatric standards, and those of the IDSA currently are undergoing revision . Most importantly, however, standards for immunization practices should be promulgated widely to all health care professionals to ensure that all segments of the population benefit from the availability of highly effective and safe vaccines.

Br J Clin Pharmacol, 2001 Mar, 51(3), 271 - 6
Sudden unexpected death in infants under 3 months of age and vaccination status- -a case-control study; Jonville-Bera AP et al.; AIMS: To determine whether DTPP+Hib vaccination (diphtheria, tetanus, pertussis, poliomyelitis +/- haemophilus) increased the risk of sudden unexpected death (SUD) in children under 3 months of age . METHODS: We conducted a multicentre case-control study in the 28 French 'SIDS Centers' . Case selection was based on death labelled sudden infant death syndrome (SIDS) of an infant aged between 30 and 90 days . Three living controls were selected, matched for sex, gestational age and born immediately after the victim in the same maternity unit . RESULTS: We identified 114 cases of SUD aged between 30 and 90 days and 341 live controls matched for age and sex and born in the same maternity unit as the case . DTPP+/-Hib immunization did not increase the risk of SUD (OR 1.08) (95% CI 0.49, 2.36) in children under 3 months of age when adjusted for sleeping position, illness in the week before death, maternal tobacco consumption, birth weight, type of mattress, breastfeeding and sex . However, low birth-weight (6.53 {2.29, 18.9}), multiple birth (5.1 {1.76, 15.13}), no breastfeeding (1.77 {1.1, 2.85}), prone sleeping position (9.8 {5, 8, 18, 9}), soft mattress (3.26 {1.69, 6.29}), recent illness (3.44 {1.84, 6.41}) and parental smoking (1.74 {1.2, 2.96}) were confirmed as risk factors in early SIDS . CONCLUSIONS: DTPP+/-Hib immunization is not a risk factor for early SUD . In this population, we found the same risk factors as described for SIDS.

FEBS Lett, 2001 Apr 6, 494(1-2), 19 - 23
Contribution of the DDDD motif of H . influenzae e (P4) to phosphomonoesterase activity and heme transport; Reilly TJ et al.; Haemophilus influenzae lipoprotein e (P4) is a member of the DDDD phosphohydrolase superfamily and mediates heme transport . Each of the aspartate residues of the signature motif is required for phosphomonoesterase activity, as none of the e (P4) single D mutants (D64A, D66A, D181N, and D185A) possessed detectable phosphomonoesterase activity . These results suggest that the signature motif is essential to the phosphomonoesterase activity of lipoprotein e (P4) . When assessed for phosphomonoesterase-dependent heme transport activity in Escherichia coli hemA strains, plasmids containing D181N and D185A retained heme transport as indicated by aerobic growth while D64A and D66A did not . We conclude that phosphomonoesterase activity is not required for heme transport.

Biochemistry, 2001 Mar 27, 40(12), 3572 - 82
Hsp70s contain a specific sulfogalactolipid binding site . Differential aglycone influence on sulfogalactosyl ceramide binding by recombinant prokaryotic and eukaryotic hsp70 family members; Mamelak D et al.; Specific 3'-sulfogalactolipid {SGL-sulfogalactosyl ceramide (SGCer) and sulfogalactosylglycerolipid (SGG)} binding is compared for hsp70s cloned from Helicobacter pylori, Haemophilus influenzae, Chlamydia trachomatis serovar E, Escherichia coli, murine male germ cells, and the hsp70-like extracellular domain within the sperm receptor from Strongylocentrotus purpuratus . This lectin activity, conserved among the different hsp70 family members, is modulated by the SGL aglycone . This is shown by differential binding to both SGC fatty acid homologues and 3'-sulfogalactolipid neoglycoproteins generated by coupling bovine serum albumin (BSA) and glycosyl ceramide acids synthesized by oxidation of the double bond of sphingosine . Eukaryotic hsp70s preferentially bound the SGCer fatty acid homologues SG(24)Cer, SG(18)Cer, and SG(20:OH)Cer, while prokaryotic hsp70s bound SG(18:1)Cer and SG(20:OH)Cer . Eukaryotic hsp70s bound SGCer-BSA and SG(24)Cer-BSA conjugates where the latter is the main constituent in SGCer-BSA, while prokaryotic hsp70s bound SG(20:OH)Cer-BSA . None of the hsp70s bound sulfogalactosyl sphingosine (SGSph) or SGSph-BSA, further demonstrating the important role of the aglycone . Although the primary SGL recognition domain of all hsp70s is conserved, we propose that aglycone organization differentially influences the interaction with the sub-site . Heterogeneous SGCer aglycone isoforms in cells and the differential in vitro binding of eukaryotic and prokaryotic hsp70s may relate to their different adhesin roles in vivo as mediators of germ cell and bacterial/host interactions, respectively.

Epidemiol Infect, 2001 Feb, 126(1), 31 - 6
The effectiveness of Haemophilus influenzae type b conjugate vaccines in a high risk population measured using immunization register data; Markey P et al.; The Northern Territory of Australia has had historically very high incidence rates of invasive Haemophilus influenzae type b disease in children less than 5 years of age, with the burden of disease greatest among Aboriginal infants less than 12 months . This study documents the impact of conjugate Hib vaccines introduced in 1993 . Immunization rates were monitored using an existing immunization register, and case finding was done retrospectively using hospital and laboratory records . Following the vaccine introduction, the incidence fell abruptly to a seventh of its pre-vaccination level, in both Aboriginal and non-Aboriginal children . The effectiveness of PRP-OMPC (PedvaxHIB) was 97.5% and the overall effectiveness of the vaccination programme was 86.3% . The study shows Hib immunization as an effective intervention while discussing continuing needs for Hib control in high risk populations . It also illustrates the benefit of immunization registers in the evaluation of immunization programmes and assessment of vaccine effectiveness.

Infect Immun, 2001 May, 69(5), 3438 - 41
Heat-inducible surface stress protein (Hsp70) mediates sulfatide recognition of the respiratory pathogen Haemophilus influenzae; Hartmann E et al.; The in vitro glycolipid binding specificity of clinical strains of nontypeable Haemophilus influenzae is altered to include sulfated glycolipids following a brief heat shock . We have constructed, expressed, and purified a recombinant protein of H . influenzae Hsp70, which showed significant specific binding to sulfated galactolipids in vitro . Furthermore, indirect immunofluorescence demonstrates that Hsp70 proteins are surface exposed in H . influenzae only after heat shock and are contained in the outer membrane protein fractions.

Infect Immun, 2001 May, 69(5), 2964 - 71
Intranasal immunization enhances clearance of nontypeable Haemophilus influenzae and reduces stimulation of tumor necrosis factor alpha production in the murine model of otitis media; Sabirov A et al.; Nontypeable Haemophilus influenzae (NTHi) is a major pathogen causing otitis media (OM) . One of the outer membrane proteins of NTHi, P6, is a common antigen to all strains and is considered a candidate for mucosal vaccine . We have previously reported that intranasal immunization with P6 and cholera toxin (CT) could induce P6-specific immunoglobulin A (IgA) antibodies in the middle ear . In the present study, we assessed the effect of intranasal immunization for the protection against NTHi-induced OM . Mice were immunized intranasally with P6 and CT as an adjuvant on days 0, 7, and 14 . Control mice were given phosphate-buffered saline (PBS) without antigen . One week after the final immunization, a suspension of live NTHi (10(7) CFU) was injected into the tympanic cavity to induce experimental OM . On days 3 and 7 after bacterial challenge, mice were killed and middle ear effusions (MEEs) were collected . All immunized mice showed elevated titers of P6-specific antibodies in MEEs . The rank order of specific antibody included, from highest to lowest levels, IgG, IgA, and IgM . In addition, immunized mice showed enhanced clearance of NTHi from the middle ear and the number of NTHi in MEEs of immunized mice was reduced by 97% on day 3 and by 92% on day 7 after bacterial challenge relative the number in the MEEs of control mice . The protective effect of intranasal immunization on the incidence of NTHi-induced experimental OM was evident on day 7 after challenge . By day 7, the number of MEEs in immunized mice was 64% less than that in control mice and the incidence of NTHi culture-positive MEEs in immunized mice was 56% less than that in control mice . Less stimulation of tumor necrosis factor alpha (TNF-alpha) production in the middle ear was evident on day 3 after challenge . Immunized mice showed lower concentrations of TNF-alpha in MEEs . These results indicate that intranasal immunization affords protection against experimental OM as evidenced by enhanced clearance of NTHi and less stimulation of TNF-alpha production in the middle ear . These findings suggest that a nasal vaccine might be useful for preventing OM.

Infect Immun, 2001 May, 69(5), 2829 - 37
Characterization of the Yersinia pestis Yfu ABC inorganic iron transport system; Gong S et al.; In Yersinia pestis, the causative agent of plague, two inorganic iron transport systems have been partially characterized . The yersiniabactin (Ybt) system is a siderophore-dependent transport system required for full virulence . Yfe is an ABC transport system that accumulates both iron and manganese . We have identified and cloned a Y . pestis yfuABC operon . The YfuABC system is a member of the cluster of bacterial ABC iron transporters that include Sfu of Serratia, Hit of Haemophilus, and Yfu of Yersinia enterocolitica . The Y . pestis KIM6+ system is most homologous to that in Y . enterocolitica, showing identities of 84% for YfuA (periplasmic binding protein), 87% for YfuB (inner membrane permease), and 75% for YfuC (ATP hydrolase) . We constructed a yfuABC promoter-lacZ fusion to examine regulation of transcription . This promoter contains a potential Fur binding sequence and is iron and Fur regulated . Significant expression from the yfuABC promoter occurred during iron-deficient growth conditions . In vitro transcription and translation of a recombinant plasmid encoding yfuABC indicates that YfuABC proteins are expressed . Escherichia coli 1017 (an enterobactin-deficient mutant) carrying this plasmid was able to grow in an iron-restrictive complex medium . We constructed a deletion encompassing the yfuABC promoter and most of yfuA . This mutation was introduced into strains with mutations in Ybt, Yfe, or both systems to examine the role of Yfu in iron acquisition in Y . pestis . Growth of the yfu mutants in a deferrated, defined medium (PMH2) at 26 and 37 degrees C failed to identify a growth or iron transport defect due to the yfu mutation . Fifty percent lethal dose studies in mice did not demonstrate a role for the Yfu system in mammalian virulence.

Clin Microbiol Rev, 2001 Apr, 14(2), 336 - 63
Bacterial infection in chronic obstructive pulmonary disease in 2000: a state-of-the-art review; Sethi S et al.; Chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death in the United States . The precise role of bacterial infection in the course and pathogenesis of COPD has been a source of controversy for decades . Chronic bacterial colonization of the lower airways contributes to airway inflammation; more research is needed to test the hypothesis that this bacterial colonization accelerates the progressive decline in lung function seen in COPD (the vicious circle hypothesis) . The course of COPD is characterized by intermittent exacerbations of the disease . Studies of samples obtained by bronchoscopy with the protected specimen brush, analysis of the human immune response with appropriate immunoassays, and antibiotic trials reveal that approximately half of exacerbations are caused by bacteria . Nontypeable Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae are the most common causes of exacerbations, while Chlamydia pneumoniae causes a small proportion . The role of Haemophilus parainfluenzae and gram-negative bacilli remains to be established . Recent progress in studies of the molecular mechanisms of pathogenesis of infection in the human respiratory tract and in vaccine development guided by such studies promises to lead to novel ways to treat and prevent bacterial infections in COPD.

Eur Respir J, 2000 Dec, 16(6), 1147 - 51
Absence of bacterial colonization of the airways after therapeutic rigid bronchoscopy without stenting; Noppen M et al.; Following airway stenting, bacterial colonization of the airways with potentially pathogenic micro-organisms occurs within 4 weeks after treatment in the majority of patients . The objective of this study was to prospectively investigate whether nonstenting therapeutic rigid bronchoscopy (using laser, cryotherapy, mechanical dilatation or debridement) is followed by airway colonization or infection . Protected specimen brush sampling of the central airways and quantitative culture were performed immediately prior to, and 4 weeks after nonstenting therapeutic rigid bronchoscopy in 20 consecutive patients with central airway lesions . Prior to therapeutic bronchoscopy, airway colonization/infection was present in nine of 20 (45%) patients . In these nine patients, 10 different potential pathogens were identified: Streptococcus pneumoniae (four cases), Pseudomonas aeruginosa (three), Haemophilus influenzae (two), and Serratia marcescens (one) . Eight of these nine patients had a history of postobstructive infections, of which three were currently being treated with antibiotics . Four weeks following therapeutic bronchoscopy, airway colonization/infection was present in five of 20 (25%) patients, each of whom had airway colonization/infection prior to bronchoscopy . In three of these five patients, the same organisms were found 4 weeks after bronchoscopy as at baseline bronchoscopy . In two of five patients new organisms were identified: one case of Streptococcus viridans and one case of Haemophilus parainfluenzae, both considered to be nonpathogens . In four of nine patients with airway colonization/infection prior to bronchoscopy, the airways were clear of micro-organisms after the procedure . The authors conclude that: 1) nonstenting therapeutic rigid bronchoscopy is not complicated by airway colonization or infection by new potential pathogens; and 2) therapeutic rigid bronchoscopy led to clearing of airway colonization/infection in almost half of the patients studied.

Clin Infect Dis, 2001 Apr 15, 32(8), 1141 - 54 Epub 2001 Mar 23.
Microbial etiology of community-acquired pneumonia in the adult population of 4 municipalities in eastern Finland; Jokinen C et al.; To determine the etiology of community-acquired pneumonia in the adult population of a defined area, specific antibody responses in paired serum samples, levels of circulating pneumococcal immune complexes in serum samples, and pneumococcal antigen in urine were measured . Samples (304 paired serum samples and 300 acute urine samples) were obtained from 345 patients > or =15 years old with community-acquired, radiologically confirmed pneumonia, which comprised all cases in the population of 4 municipalities in eastern Finland during 1 year . Specific infecting organisms were identified in 183 patients (including 49 with mixed infection), as follows: Streptococcus pneumoniae, 125 patients; Haemophilus influenzae, 12; Moraxella catarrhalis, 8; chlamydiae, 37 (of which, Chlamydia pneumoniae, 30); Mycoplasma pneumoniae, 30; and virus species, 27 . The proportion of patients with pneumococcal infections increased and of those with Mycoplasma infections decreased with age, but for each age group, the etiologic profile was similar among inpatients and among outpatients . S . pneumoniae was the most important etiologic agent . The annual incidence of pneumococcal pneumonia per 1000 inhabitants aged > or =60 years was 8.0.

J Clin Microbiol, 2001 Apr, 39(4), 1553 - 8
Simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in suspected cases of meningitis and septicemia using real-time PCR; Corless CE et al.; A single-tube 5' nuclease multiplex PCR assay was developed on the ABI 7700 Sequence Detection System (TaqMan) for the detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae from clinical samples of cerebrospinal fluid (CSF), plasma, serum, and whole blood . Capsular transport (ctrA), capsulation (bexA), and pneumolysin (ply) gene targets specific for N . meningitidis, H . influenzae, and S . pneumoniae, respectively, were selected . Using sequence-specific fluorescent-dye-labeled probes and continuous real-time monitoring, accumulation of amplified product was measured . Sensitivity was assessed using clinical samples (CSF, serum, plasma, and whole blood) from culture-confirmed cases for the three organisms . The respective sensitivities (as percentages) for N . meningitidis, H . influenzae, and S . pneumoniae were 88.4, 100, and 91.8 . The primer sets were 100% specific for the selected culture isolates . The ctrA primers amplified meningococcal serogroups A, B, C, 29E, W135, X, Y, and Z; the ply primers amplified pneumococcal serotypes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10A, 11A, 12, 14, 15B, 17F, 18C, 19, 20, 22, 23, 24, 31, and 33; and the bexA primers amplified H . influenzae types b and c . Coamplification of two target genes without a loss of sensitivity was demonstrated . The multiplex assay was then used to test a large number (n = 4,113) of culture-negative samples for the three pathogens . Cases of meningococcal, H . influenzae, and pneumococcal disease that had not previously been confirmed by culture were identified with this assay . The ctrA primer set used in the multiplex PCR was found to be more sensitive (P < 0.0001) than the ctrA primers that had been used for meningococcal PCR testing at that time.

Genome Res, 2001 Apr, 11(4), 566 - 84
A comparative genomics approach to prediction of new members of regulons; Tan K et al.; Identifying the complete transcriptional regulatory network for an organism is a major challenge . For each regulatory protein, we want to know all the genes it regulates, that is, its regulon . Examples of known binding sites can be used to estimate the binding specificity of the protein and to predict other binding sites . However, binding site predictions can be unreliable because determining the true specificity of the protein is difficult because of the considerable variability of binding sites . Because regulatory systems tend to be conserved through evolution, we can use comparisons between species to increase the reliability of binding site predictions . In this article, an approach is presented to evaluate the computational predictions of regulatory sites . We combine the prediction of transcription units having orthologous genes with the prediction of transcription factor binding sites based on probabilistic models . We augment the sets of genes in Escherichia coli that are expected to be regulated by two transcription factors, the cAMP receptor protein and the fumarate and nitrate reduction regulatory protein, through a comparison with the Haemophilus influenzae genome . At the same time, we learned more about the regulatory networks of H . influenzae, a species with much less experimental knowledge than E . coli . By studying orthologous genes subject to regulation by the same transcription factor, we also gained understanding of the evolution of the entire regulatory systems.

Vaccine, 2001 Apr 6, 19(20-22), 2924 - 31
Safety and immunogenicity of three lots of meningococcal serogroup C conjugate vaccine administered at 2, 3 and 4 months of age; Bramley JC et al.; The reactogenicity and immunogenicity of meningococcal serogroup C conjugate (MenC) vaccine was assessed in 322 infants vaccinated at 2, 3, and 4 months of age, with concomitant administration of mixed diphtheria-tetanus-whole-cell pertussis vaccine and Haemophilus influenzae type b conjugate vaccine (DTwP-Hib) and oral polio vaccine . All infants in whom post-vaccination meningococcal C anticapsular IgG levels were assayed (n = 265) attained > or = 2 microg ml(-1) . Serum bactericidal titres were assayed for a proportion of subjects (n = 171), 98% of whom obtained a reciprocal titres > or = 8 . Local reactions were less frequent at the MenC injection site than at the DTP-Hib site . Systemic events were frequent, but consistent with established DTwP-Hib experience . The study demonstrates that MenC vaccine is immunogenic and well tolerated in infants at manufacturing scale production levels.

Commun Dis Public Health, 2000 Dec, 3(4), 291 - 4
Outcome of medical screening of Kosovan refugees in Ireland: 1999; Smith A et al.; In March 1999 armed conflict broke out in Kosova and about 900,000 ethnic Albanians were displaced . We reviewed the health care offered to the 945 Kosovan refugees who arrived in Ireland in 1999, which included screening for tuberculosis (TB) and hepatitis B . On arrival in Ireland 540 refugees had already received oral polio vaccine (57%), 512 diphtheria, tetanus, and acellular pertussis or diphtheria and tetanus vaccine (54%), 310 BCG (33%), 207 measles, mumps, and rubella vaccine (22%) and 60 Haemophilus influenzae type b (6%) . Twelve refugees were diagnosed with TB . Twenty-six refugees were HBsAg positive (3%) and 168 were anti-HBcAg positive (18%) . Organised screening of Kosovan refugees on a voluntary basis (uptake > 95%) revealed low percentages who had been immunised and relatively high rates of TB and hepatitis B . The provision of optimum immunisation, screening, and treatment services to address these issues requires substantial staffing and financial resources.

Vet Microbiol, 2001 May 3, 80(1), 53 - 61
Virulence of Pasteurella multocida recA mutants; Cardenas M et al.; In order to determine the role of the RecA protein in the virulence of Pasteurella multocida, a recA mutant was constructed and used in studies of virulence and competition in relation to wild-type strain . To achieve this, firstly, the recA gene was isolated and sequenced, showing an Escherichia coli-like SOS box and encoding a protein of 354 amino acids which has the closest identity with the Haemophilus influenzae RecA protein . Further, the recA mutant was constructed, by inactivating this gene by single recombination of a suicide plasmid containing an internal region of the P . multocida recA gene, and shown to be more sensitive to UV radiation than the parental strain . The P . multocida mutant was slightly attenuated in virulence, as indicated by the LD(50), the time of death of infected animals, and a failure to compete with the wild-type strain in mixed infections . Compared to the parent strain, the mutant had a similar growth rate but a longer lag phase . These data suggest that the diminished virulence of the recA mutant as well as its failure in competition were more a consequence of the long lag phase rather than a direct effect of the inactivation of the recA gene on genes involved in virulence.

Biochimie, 2001 Feb, 83(2), 235 - 41
H-NS and H-NS-like proteins in Gram-negative bacteria and their multiple role in the regulation of bacterial metabolism; Bertin P et al.; In Escherichia coli, the H-NS protein plays an important role in the structure and the functioning of bacterial chromosome . A homologous protein has also been identified in several enteric bacteria and in closely related organisms such as Haemophilus influenzae . To get information on their structure and their function, we identified H-NS-like proteins in various microorganisms by different procedures . In silico analysis of their amino acid sequence and/or in vivo experiments provide evidence that more than 20 proteins belong to the same class of regulatory proteins . Moreover, large scale technologies demonstrate that, at least in E . coli, the loss of motility in hns mutants results from a lack of flagellin biosynthesis, due to the in vivo repression of flagellar gene expression . In contrast, several genes involved in adaptation to low pH are strongly induced in a H-NS deficient strain, resulting in an increased resistance to acidic stress . Finally, expression profiling and phenotypic analysis suggest that, unlike H-NS, its paralogous protein StpA does not play any role in these processes.

Eur J Biochem, 2001 Apr, 268(7), 2148 - 59
A new structural type for Haemophilus influenzae lipopolysaccharide . Structural analysis of the lipopolysaccharide from nontypeable Haemophilus influenzae strain 486; Mansson M et al.; Structural elucidation of the sialylated lipopolysaccharide (LPS) of non-typeable Haemophilus influenzae (NTHi) strain 486 has been achieved by the application of high-field NMR techniques and ESI-MS along with composition and linkage analyses on O-deacylated LPS and oligosaccharide samples . It was found that the LPS contains the common element of H . influenzae, L-alpha-D-Hepp-(1-->2)-{PEtn-->6}-L-alpha-D-Hepp-(1-->3)-{beta-D-Glcp-(1-->4)}-L-alpha-D-Hepp-(1-->5)-{PPEtn-->4}-alpha-Kdop-(2-->6)-Lipid A, but instead of glycosyl substitution of the terminal heptose residue (HepIII) at the O2 position observed in other H . influenzae strains, HepIII is chain elongated at the O3 position by either lactose or sialyllactose (i.e . alpha-Neu5Ac-(2-->3)-beta-D-Galp-(1-->4)-beta-D-Glcp) . The LPS is substituted by an O-acetyl group linked to the O2 position of HepIII and phosphocholine (PCho) which was located at the O6 position of a terminal alpha-D-Glcp residue attached to the central heptose, a molecular environment different from what has been reported earlier for PCho . In addition, minor substitution by O-linked glycine to the LPS was observed . By investigation of LPS from a lpsA mutant of NTHi strain 486, it was demonstrated that the lpsA gene product also is responsible for chain extension from HepIII in this strain . The involvement of lic1 in expression of PCho was established by investigation of a lic1 mutant of NTHi strain 486.

Microb Pathog, 2001 Mar, 30(3), 157 - 66
The impact of Haemophilus ducreyi cytolethal distending toxin on cells involved in immune response; Svensson LA et al.; The Haemophilus ducreyi cytolethal distending toxin (HdCDT) induces cell cycle arrest and thereby inhibits cell proliferation of many cultured mammalian cell-lines . We investigated the effect of HdCDT on circulating human hematopoietic cells, including T- and B-cells, monocytes and polymorphonuclear cells (PMN) . Lymphocytes were stimulated with T- and B-cell specific mitogens, whereas monocytes and PMN with endotoxin . HdCDT inhibited the mitogen-induced proliferation of T-cells in a dose-dependent manner as assayed by {(3)H}-thymidine incorporation and MTT assays . Similarly to T-cells, HdCDT also inhibited the proliferation of B-cells and consequently the immunoglobulin production, measured by ELISPOT and ELISA assays . In contrast, the HdCDT did not affect monocytes or PMN, as measured by MTT assay . The TNF-alpha production by monocytes and the phagocytic ability of PMN were neither affected . The monocytic cell line THP-1 was, however, sensitive to the toxin, seen as a reduction of proliferation and viability after exposure to HdCDT . In conclusion, exposure to HdCDT significantly affects the proliferation and other biological activities of stimulated human T- and B-cells, while circulating monocytes and PMN are not sensitive to HdCDT . The sensitivity of cells of the acquired immune system to HdCDT may hamper specific host response to H . ducreyi and contribute to persistence of chancroid lesions .

Microb Pathog, 2001 Mar, 30(3), 121 - 7
Modulating effects of mucoregulating drugs on the attachment of Haemophilus influenzae; Ndour CT et al.; Non-typable Haemophilus influenzae (NTHI) is one of the three major pathogens implicated in human respiratory infections . The ability to attach with pharyngeal epithelial cells is an important factor for infection and virulence . In the present study we describe the effects of two mucoregulating drugs, S-carboxymethylcysteine (S-CMC) and ambroxol, on the attachment of NTHI to pharyngeal epithelial cells . There was a significant (P < 0.0001, < 0.001 and <0.01) decrease of attachment (8.8 +/ 2.4, 9.2+/-2.5 and 15.4 +/- 5.7 bactreria/cell) compared with the control (17.5 +/- 2.9, 15.5 +/- 3.1 and 18.8 +/- 6.8 bacteria/cell) after cells were treated wth S-CMC at a dose of 100, 10 and 1 microg/ml . After attachment assay, cells treated with S-CMC (100 microg/ml) showed a significant decrease (P < 0.01) of attached bacteria (3.1 +/- 0.8 bacteria/cell) compared with the control (5.9 +/- 1.8 bacteria/cell) . Treatment of cells with ambroxol did not influence bacterial attachment . By scanning electron microscopic observation it was found that NTHI attaches to the surface elevations (microplicae) of human pharyngeal epithelial cells . Atomic force microscopic observation revealed that the surface potential of microplicae decreased significantly in cells treated with S-CMC compared with the untreated control cells . As bacteria with negative surface charge attach to the positively charged domain, i.e . microplicae of human pharyngeal epithelial cells, this study suggests that the decrease of attachment of NTHI with epithelial cells after treatment with S-CMC was possibly due to the decrease of surface charge . This study suggests that S-CMC decreases the episodes of respiratory infections in patients with respiratory diseases both by inhibiting the attachment of bacteria to the upper respiratory tract, and by detaching the adherent one .

Microb Pathog, 2001 Mar, 30(3), 111 - 20
Expression of molecular markers for bone formation increases during experimental acute otitis media; Melhus A et al.; Bony tissues are integral parts of the function of the middle ear and the protection of adjacent vital structures . To explore the reaction of middle ear bone to acute otitis media, rats were challenged with Streptococcus pneumoniae and Haemophilus influenzae . Local changes were monitored for up to 1 month . After reverse transcription, competitive polymerase chain reaction was used to determine the expression levels of two molecular markers of bone formation, osteocalcin and procollagen I, and the two cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha, in the bone . Middle ear bone responded rapidly to bacterial challenge, and the reaction depended upon the causative agent . On day 1, IL-6 and TNF-alpha transcripts were detected in the bone from all middle ears . After a short period of decreased expression of osteocalcin, during which the otitis diagnosis could not be made clinically, the levels of bone formation markers increased dramatically . The maximum levels of these markers were reached on days 6 and 14 for animals challenged with H . influenzae and pneumococci, respectively . Infections induced by pneumococci had a longer duration, and after the initial phase the production of osteocalcin and procollagen transcript were significantly higher in the pneumococcus-infected animals . The results indicate that even in an uncomplicated infection, the bone of the bulla reacts to an acute otitis media with a short period of inhibited osteoblast activity followed by a longer period of new bone formation .

Am Fam Physician, 2001 Feb 1, 63(3), 499 - 506, 508
Detection, education and management of the asplenic or hyposplenic patient; Brigden ML; Fulminant, potentially life-threatening infection is a major long-term risk after splenectomy or in persons who are functionally hyposplenic as a result of various systemic conditions . Most of these infections are caused by encapsulated organisms such as pneumococci, Haemophilus influenzae and meningococci . A splenectomized patient is also more susceptible to infections with intraerythrocytic organisms such as Babesia microti and those that seldom affect healthy people, such as Capnocytophaga canimorsus . Most patients who have lost their spleens because of trauma are aware of their asplenic condition, but some older patients do not know that they are asplenic . Other patients may have functional hyposplenism secondary to a variety of systemic diseases ranging from celiac disease to hemoglobinopathies . The identification of Howell-Jolly bodies on peripheral blood film is an important clue to the diagnosis of asplenia or hyposplenia . Management of patients with these conditions includes a combination of immunization, antibiotic prophylaxis and patient education . With the increasing prevalence of antibiotic-resistant pneumococci, appropriate use of the pneumococcal vaccine has become especially important.

Arch Pediatr, 2001 Mar, 8(3), 316 - 20
{Secondary effects of vaccinations}; Ovetchkine P; The adverse effects of vaccines include local reactions and systemic symptoms or illnesses . Local reactions are frequent, most often presenting as transient pain, redness, edema and/or nodule . Fever of short duration is the main systemic symptom, generally occurring within 24-48 hours following vaccination . Some vaccines have recognized specific adverse effects such as thrombocytopenic purpura for the measles-mumps-rubella vaccine, and febrile convulsions for the pertussis vaccine . Hepatitis B vaccine and Haemophilus influenzae type b vaccine have been respectively suspected to be responsible for neurological demyelinating disease and insulin-dependent diabetes mellitus, but large-scale epidemiological studies have failed to confirm these allegations.

J Microbiol Immunol Infect, 2000 Dec, 33(4), 237 - 40
Mastoiditis: a disease often overlooked by pediatricians; Cheng MF et al.; Although mastoiditis can be a life threatening disease, clinicians often overlook it because it is uncommon . We reviewed the presentation and management of all children younger than 15 years of age with the discharge diagnosis of mastoiditis in our hospital from January 1994 through December 1999 . Nineteen patients that fulfilled the case definition were included . The most common clinical presentation in this series was fever . More specific findings, such as otorrhea, postauricular pain, swelling, and redness of mastoid could be found in less than half of these patients . Only two patients had characteristic physical findings, and mastoiditis was diagnosed in only three patients upon admission . Plain radiographic evidence of mastoiditis was usually not apparent early in the course . In this series, the majority of patients were diagnosed by computed tomography (CT) scans . The present study demonstrates that mastoiditis most commonly presents without a clearly diagnostic set of physical examination and laboratory findings . Mastoiditis should be considered in patients with otitis media or with fever of unknown origin (FUO) . The empirical antibiotic treatment should cover organisms commonly found in acute otitis media (AOM), including Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis.

J Antimicrob Chemother, 2001 Apr, 47(4), 475 - 7
The relationship between trends in macrolide use and resistance to macrolides of common respiratory pathogens; Cizman M et al.; The correlation between increased macrolide consumption and the resistance of Streptococcus pyogenes, Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis to macrolides in Slovenia from 1994 to 1999 was evaluated . The outpatient consumption of macrolides increased from 1.89 to 3.84 defined daily doses (DDD)/1000 inhabitants/day during the observation period . This increase in macrolide consumption was paralleled by a steady increase in macrolide resistance in S . pyogenes (from 0 to 7.4%, r = 0.90, P = 0.014) and upper respiratory S . pneumoniae isolates (from 0 to 9%, r = 0.82, P = 0.044) . In other pathogens studied, no significant increase was detected.

J Antimicrob Chemother, 2001 Apr, 47(4), 471 - 3
Antibacterial activity of telithromycin (HMR 3647) in relation to in vitro simulated human plasma kinetics; Bonnefoy A et al.; Telithromycin (HMR 3647) is a ketolide suitable for the treatment of respiratory infections . The aim of this study was to demonstrate its antibacterial efficacy against an erythromycin-susceptible Staphylococcus aureus, an erythromycin-resistant Streptococcus pneumoniae and Haemophilus influenzae . The free serum concentrations of telithromycin, produced by repeated oral administration of 800 mg to adults for 10 days, was simulated in an in vitro system . The ketolide displayed bacteriostatic activity against all three strains tested . This study supported the observation that an 800 mg po dose of telithromycin demonstrated antibacterial efficacy against respiratory tract pathogens.

Emerg Infect Dis, 2001 Jan-Feb, 7(1), 92 - 9
Active bacterial core surveillance of the emerging infections program network; Schuchat A et al.; Active Bacterial Core surveillance (ABCs) is a collaboration between the Centers for Disease Control and Prevention and several state health departments and universities participating in the Emerging Infections Program Network . ABCs conducts population-based active surveillance, collects isolates, and performs studies of invasive disease caused by Streptococcus pneumoniae, group A and group B Streptococcus, Neisseria meningitidis, and Haemophilus influenzae for a population of 17 to 30 million . These pathogens caused an estimated 97,000 invasive cases, resulting in 10,000 deaths in the United States in 1998 . Incidence rates of these pathogens are described . During 1998, 25% of invasive pneumococcal infections in ABCs areas were not susceptible to penicillin, and 13.3% were not susceptible to three classes of antibiotics . In 1998, early-onset group B streptococcal disease had declined by 65% over the previous 6 years . More information on ABCs is available at ABCs specimens will soon be available to researchers through an archive.

Postgrad Med J, 2001 Apr, 77(906), 261 - 2
It could only happen to a doctor--Haemophilus aphrophilus septicaemia complicated by a prevertebral infection after dental work; Poullis A et al.; A 53 year old man presented with severe neck pain and a flu-like illness; he had recently returned from Sri Lanka and had had dental treatment six days before illness onset . Blood culture showed infection by Haemophilus aphrophilus . Magnetic resonance imaging was performed and exploratory surgery undertaken . The prevertebral cervical fascia was inflamed but no abscess identified . He was treated with antibiotics and made an uneventful recovery.

J Health Popul Nutr, 2000 Dec, 18(3), 131 - 8
Age- and cause-specific childhood mortality in Lombok, Indonesia, as a factor for determining the appropriateness of introducing Haemophilus influenzae type b and pneumococcal vaccines; Nelson CM et al.; Using age and cause-specific childhood mortality in Lombok, Indonesia, as a factor for determining the appropriateness of introducing Haemophilus influenzae type b (Hib) and pneumococcal vaccines, the study describes a cross-sectional, hamlet-level mortality survey in 40 of 305 villages in Lombok Island, Indonesia . Causes of death were assessed with a standardized verbal-autopsy questionnaire . One thousand four hundred ninety-nine births and 141 deaths occurring among children aged less than 2 years were identified, with 43% of deaths occurring during the first 2 months of life . The infant mortality rate was 89 (95% CI: 75, 104) per 1,000 live-births . All mortality rates are reported per 1,000 live-births . To examine children whose deaths could potentially have been prevented through vaccination with Hib or pneumococcal vaccine, deaths due to acute respiratory infection (ARI) and central nervous system (CNS) infections among children, aged 2-23 months, were analyzed . ARI and CNS infections caused 58% (mortality rate: 31 per 1,000 live-births; 95% CI: 23, 41) and 17% (mortality rate: 9 per 1,000 live-births; 95% CI: 5, 16), respectively, of all deaths within this age group . Between the ages of 2 and 23 months, 5% of all babies born alive died of ARI, and another 1% died of CNS infections . Our results indicate that current efforts to reduce childhood mortality should focus on reducing ARI and meningitis . These efforts should include evaluating the impact of Hib and pneumococcal vaccines within the routine Expanded Programme on Immunization system.

Immunogenetics, 2001 Feb, 53(1), 22 - 30
Distribution of human kappa locus IGKV2-29 and IGKV2D-29 alleles in Swedish Caucasians and Hong Kong Chinese; Padyukov L et al.; Polymorphism in the IGKV2-29 gene was shown to decrease the recombination frequency in B cells and to be important for immune responses to Haemophilus influenzae type b polysaccharide . By using the combination of PCR and restriction enzyme mapping, the distribution of IGKV2D-29 and IGKV2-29 gene alleles was estimated in two geographically and ethnically different groups . We found that V2D-29*01 homozygous individuals were most common in Swedish Caucasians (82%), but less common in the Chinese population of Hong Kong (28%) . The homozygous V2D-29*02 genotype was found in 19% Chinese, but only in one Caucasian (1%) . The frequency of the heterozygous V2D-29*01/V2D-29*02 genotype was also higher in the Chinese population (46%) compared with the Caucasians (7%) . V2-29*01 homozygosity was more frequent among Caucasians (85%) than among Chinese (19%) . In contrast, homozygous V2-29*02 individuals were over-represented in the Chinese population (18%), whereas only one was found among Caucasians (1%) . Heterozygous V2-29*01/V2-29*02 individuals were also more common in the Chinese (63%) than the Caucasian (15%) population . Most Caucasians had the combination of V2D-29*01/V2D-29*01+V2-29*01/V2-29*01 (74%), while the most common genotype for Chinese was V2D-29*01/V2D-29*02+ V2-29*01/V2-29*02 (41%) . Analysis of the association of V2D-29*02 and V2-29*02 alleles demonstrated a high degree of linkage, as for V2D-29*01 with V2-29*01 . These data show a significant difference in the distribution of IGKV2D-29 and IGKV2-29 alleles among Swedish Caucasians and Hong Kong Chinese . This may help to explain differences in the occurrence of H . influenzae type b infection in the two populations . Evaluated methods for IGKV2D-29 and IGKV2-29 allele detection can be used for the screening allele polymorphisms in other particular patient groups.

Indian J Cancer, 2000 Mar, 37(1), 15 - 22
Production of monoclonal antibodies to a tumor--associated antigen by spontaneous cell fusion; Subbiah K et al.; Spontaneous cell fusion induced by the bacterium Haemophilus paragallinarum has been recently reported as an alternative technique to generate hybridomas producing monoclonal antibody (mAb) . In order to investigate the advantages of this technique to produce anti-tumor monoclonal antibodies we performed comparative experiments between H . paragallinarum induced spontaneous cell fusion and polyethylene glycol (PEG) mediated fusion . Hybridomas producing monoclonal antibodies to an experimental murine lymphoma antigen, the Dalton's lymphoma associated antigen (DLAA) were generated and their sensitivity and specificity were ascertained . The spontaneous fusion yielded more number of stable and specific hybridomas than PEG mediated fusion . The results suggest the advantage of H . paragalinarum induced cell fusion for the simplified production of specific antitumor monoclonal antibodies.

Kansenshogaku Zasshi, 2001 Feb, 75(2), 124 - 32
{Pathogenic bacteria in the nasal vestivulum of children with acute respiratory tract infection}; Kakinohana S et al.; The isolation frequency of pathogenic bacteria for acute respiratory infection (ARI) in the pharynx and nasal vestivulum was investigated . Age group-matched children with or without ARI including 109 individuals in each group were examined . Any of the organisms, which are widely regarded as the pathogens causing ARI such as Haemophilus influenzae, Streptococcus pneumoniae, beta-haemolytic Streptococcus, Staphylococcus aureus, and Moraxella catarrhalis, were isolated from 91% of the patient group and from 77% of the healthy group . The isolation frequency of S . pneumoniae in the nasal vestivulum of the patient group was outstanding . The healthy carrier rates of S . pneumoniae in the pharynx and nasal vestivulum were 9% and 8%, respectively . Whereas the isolation frequencies from the patient group were 7% and 28%, respectively . alpha-haemolytic Streptococci except S . pneumoniae revealed different tendency from S . pneumoniae . These organisms were almost always isolated from their pharynx but rarely isolated from the nasal vestivulum . The isolation frequency of H . influenzae from the pharynx of the patient group was 41%, which was slightly higher than 34% in the healthy group, but the difference was statistically not significant . H . influenzae was not isolated from the nasal vestivulum of the healthy group, nevertheless it was isolated from 25% of the patient group . The isolation of H . influenzae from the nasal vestivulum may have some important information about ARI . S . aureus was isolated from the pharynx with higher rate than the nasal vestivulum in both groups, and moreover, the isolation frequency of S . aureus in the healthy group was higher than the patient group . It means that the diagnosis of staphylococcal infection should be made very carefully . Considering the results of this study, it could be said that bacteriologic examination of the specimens from nasal vestivulum is valuable to determine S . pneumoniae and H . influenzae as the etiologic agents of ARI.

Arch Dis Child, 2001 Apr, 84(4), 332 - 6
Procalcitonin in children admitted to hospital with community acquired pneumonia; Moulin F et al.; AIMS: To assess the sensitivity, specificity, and predictive value of procalcitonin (PCT) in differentiating bacterial and viral causes of pneumonia . METHODS: A total of 72 children with community acquired pneumonia were studied . Ten had positive blood culture for Streptococcus pneumoniae and 15 had bacterial pneumonia according to sputum analysis (S pneumoniae in 15, Haemophilus influenzae b in one) . Ten patients had Mycoplasma pneumoniae infection and 37 were infected with viruses, eight of whom had viral infection plus bacterial coinfection . PCT concentration was compared to C reactive protein (CRP) concentration and leucocyte count, and, if samples were available, interleukin 6 (IL-6) concentration . RESULTS: PCT concentration was greater than 2 microg/l in all 10 patients with blood culture positive for S pneumoniae; in eight of these, CRP concentration was above 60 mg/l . PCT concentration was greater than 1 microg/l in 86% of patients with bacterial infection (including Mycoplasma and bacterial superinfection of viral pneumonia) . A CRP concentration of 20 mg/l had a similar sensitivity but a much lower specificity than PCT (40% v 86%) for discriminating between bacterial and viral causes of pneumonia . PCT concentration was significantly higher in cases of bacterial pneumonia with positive blood culture whereas CRP concentration was not . Specificity and sensitivity were lower for leucocyte count and IL-6 concentration . CONCLUSIONS: PCT concentration, with a threshold of 1 microg/l is more sensitive and specific and has greater positive and negative predictive values than CRP, IL-6, or white blood cell count for differentiating bacterial and viral causes of community pneumonia in untreated children admitted to hospital as emergency cases.

Am J Health Syst Pharm, 2001 Mar 1, 58(5), 379 - 88
Moxifloxacin: clinical efficacy and safety; Culley CM et al.; The activity, pharmacokinetics, pharmacodynamics, efficacy, safety, drug interactions, and dosage and administration of moxifloxacin are reviewed . Moxifloxacin is an oral 8-methoxyquinolone antimicrobial approved in December 1999 for use in the treatment of acute bacterial sinusitis, acute bacterial exacerbations of chronic bronchitis, and community-acquired pneumonia . This fluoroquinolone is active against common community-acquired respiratory pathogens (Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis), atypical pathogens, and many anaerobes . Moxifloxacin has an absolute bioavailability of 90% after oral administration and a mean elimination half-life of 12 hours . The drug is not a substrate or inhibitor of the hepatic cytochrome P-450 isoenzyme system thereby avoiding many potential drug interactions . Moxifloxacin has limited phototoxic potential . In clinical trials, moxifloxacin had clinical success rates of 88-97% and bacteriologic eradication rates of 90-97% . Reported adverse effects were primarily gastrointestinal (nausea, diarrhea) and were mild to moderate in severity . Moxifloxacin prolongs the QT interval by a mean + S.D . of 6 +/- 26 milliseconds above baseline and should be used with caution in patients with proarrhythmic conditions and avoided in patients receiving antiarrhythmia agents, such as quinidine, procainamide, amiodarone, and sotalol . The standard oral dosage is 400 mg once a day . Dosage adjustment is unnecessary in patients with renal dysfunction or mild to moderate hepatic dysfunction . Moxifloxacin is a safe and effective antimicrobial that will be useful for treating acute sinusitis, acute bacterial exacerbations of chronic bronchitis, and community-acquired pneumonia.

Vaccine, 2001 Mar 21, 19(17-19), 2522 - 6
Programmed inflammatory processes induced by mucosal immunisation; Foxwell AR et al.; Inflammation is essential to repair tissue damaged by physical, microbial or allergic mechanisms . Inappropriately zealous responses lead to destructive pathology or chronic disease cycles, whereas ideal outcomes are associated with complete and rapid restoration of tissue structure and function . The establishment of a rodent model investigating the different immune responses to non-typeable Haemophilus influenzae infection in both the lung and the ear indicate an ability to clear bacteria and reduce inflammation following mucosal immunisation . Lung histochemistry, upregulaion of macrophages and polymorphonuclear neutrophils, recruitment of gammadelta(+) and CD8(+) T cells, cytokine levels and depletion studies all support the hypothesis that mucosal immunisation facilitates control of the immune response resulting in enhanced bacterial clearance and programming of inflammation which limits damage and promotes the rapid restoration of structural normality.

Vaccine, 2001 Mar 21, 19(17-19), 2319 - 22
Conjugates and reverse vaccinology to eliminate bacterial meningitis; Rappuoli R; Bacterial meningitis is caused mainly by Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis . Haemophilus influenzae type b vaccines have been extremely successful in eradicating the disease from those countries where the vaccine has been introduced . The recent licensure of the conjugated pneumococcal vaccine suggests that this pathogen also will be soon controlled . Consequently, if we succeed in developing effective vaccines against meningococcus, this will enable us to eliminate bacterial meningitis . The global elimination of bacterial meningitis is a goal which, if appropriate resources are applied, can be reached within the first fifteen years of the 21st century.

Vaccine, 2001 Mar 21, 19(17-19), 2280 - 5
Clinical relevance of lower Hib response in DTPa-based combination vaccines; Poolman J et al.; Combination vaccines are essential to enable administration of all the required antigens in routine infant immunisation schedules at any single visit . Some combinations of diphtheria-tetanus-acellular pertussis (DTPa) with Haemophilus influenzae type b (Hib) conjugate vaccines have been shown to result in lower Hib titres than when Hib is administered separately . While confirming that a primary series with a DTPa-HBV-IPV/Hib combination gives lower antibody levels than separate Hib conjugates, we show that the nature (isotype and IgG subclasses) and function (avidity and opsonic activity) of the antibodies are the same, and immunologic memory is induced . It is likely therefore that the DTPa-HBV-IPV/Hib combination will be efficacious against Hib disease.

Antimicrob Agents Chemother, 2001 Apr, 45(4), 1058 - 64
Peptide deformylase as an antibacterial drug target: target validation and resistance development; Apfel CM et al.; New inhibitors of peptide deformylase (PDF) which are very potent against the isolated enzyme and show a certain degree of antibacterial activity have recently been synthesized by our group . Several lines of experimental evidence indicate that these inhibitors indeed interfere with the target enzyme in the bacterial cell . (i) The inhibition of Escherichia coli growth could be counteracted by overexpression of PDF from different organisms, including E . coli, Streptococcus pneumoniae, and Haemophilus influenzae . Conversely, reduced expression of PDF in S . pneumoniae resulted in an increased susceptibility to the inhibitors . (ii) Proteome analysis on two-dimensional gels revealed a shift for many proteins towards lower pI in the presence of PDF inhibitors, as would be expected if the proteins still carry their N-formyl-Met terminus . (iii) PDF inhibitors show no antimicrobial activity against E . coli under conditions that make growth independent of formylation and deformylation . The antibacterial activity in E . coli was characterized as bacteriostatic . Furthermore, the development of resistance in E . coli was observed to occur with high frequency (10(-7)) . Resistant mutants show a reduced growth rate, and DNA sequence analysis revealed mutations in their formyl transferase gene . Taking all these aspects into account, we conclude that PDF may not be an optimal target for broad-spectrum antibacterial agents.

Acta Otorhinolaryngol Belg, 2001, 55(1), 83 - 6
Turnover of Haemophilus influenzae isolates in otitis-prone children; van Kempen M et al.; Arbitrarily primed PCR with primer ERIC2 and RAPD Ready-to-Go beads was applied to study the epidemiology of non-typable Haemophilus influenzae (NTHI) isolated from the nasopharynx of children with recurrent otitis media (otitis-prone children) . Thirty-five otitis-prone children (OP-children) were included . Three pairs of siblings were identified in the study population . This study is part of a large prospective multicentric trial investigating the efficacy of a new conjugated pneumococcal vaccine in OP-children (OMAVAX trial) . During a 2-year study period, NTHI strains were isolated from nasopharyngeal swabs and, when possible, middle ear aspirates were collected in OP-children between 1 and 6 years of age . In 20 out of the 35 children, 48 H . influenzae isolates were obtained simultaneously from different sites (left and/or right ear and/or nasopharynx) of the same child as well as from siblings or during initial and follow-up visits of the same child . Subsequent genotyping indicated substantial genetic diversity among the H . influenzae isolates studied, since for a total of 48 isolates in 20 OP-children, 29 different genotypes were observed . Simultaneous isolation for different sampling sites (ear or nasopharynx) as well as for siblings resulted mostly in identical fingerprints . Longitudinal follow-up of H . influenzae isolates in the nasopharynx almost always resulted in different genotypes . We can therefore conclude that both cross colonization (between sampling sites within the same patient and between siblings) and turnover of H . influenzae isolates are high in OP-children.

Ann Intern Med, 2001 Mar 20, 134(6), 498 - 505
Principles of appropriate antibiotic use for acute rhinosinusitis in adults: background; Hickner JM et al.; The following principles of appropriate antibiotic use for adults with acute rhinosinusitis apply to the diagnosis and treatment of acute maxillary and ethmoid rhinosinusitis in adults who are not immunocompromised.1 . Most cases of acute rhinosinusitis diagnosed in ambulatory care are caused by uncomplicated viral upper respiratory tract infections . 2 . Bacterial and viral rhinosinusitis are difficult to differentiate on clinical grounds . The clinical diagnosis of acute bacterial rhinosinusitis should be reserved for patients with rhinosinusitis symptoms lasting 7 days or more who have maxillary pain or tenderness in the face or teeth (especially when unilateral) and purulent nasal secretions . Patients with rhinosinusitis symptoms that last less than 7 days are unlikely to have bacterial infection, although rarely some patients with acute bacterial rhinosinusitis present with dramatic symptoms of severe unilateral maxillary pain, swelling, and fever.3 . Sinus radiography is not recommended for diagnosis in routine cases . 4 . Acute rhinosinusitis resolves without antibiotic treatment in most cases . Symptomatic treatment and reassurance is the preferred initial management strategy for patients with mild symptoms . Antibiotic therapy should be reserved for patients with moderately severe symptoms who meet the criteria for the clinical diagnosis of acute bacterial rhinosinusitis and for those with severe rhinosinusitis symptoms-especially those with unilateral facial pain-regardless of duration of illness . For initial treatment, the most narrow-spectrum agent active against the likely pathogens, Streptococcus pneumoniae and Haemophilus influenzae, should be used.

Thorax, 2001 Apr, 56(4), 296 - 301
Study of community acquired pneumonia aetiology (SCAPA) in adults admitted to hospital: implications for management guidelines; Lim WS et al.; BACKGROUND: Since the last British study of the microbial aetiology of community acquired pneumonia (CAP) about 20 years ago, new organisms have been identified (for example, Chlamydia pneumoniae), new antibiotics introduced, and fresh advances made in microbiological techniques . Pathogens implicated in CAP in adults admitted to hospital in the UK using modern and traditional microbiological investigations are described . METHODS: Adults aged 16 years and over admitted to a teaching hospital with CAP over a 12 month period from 4 October 1998 were prospectively studied . Samples of blood, sputum, and urine were collected for microbiological testing by standard culture techniques and new serological and urine antigen detection methods . RESULTS: Of 309 patients admitted with CAP, 267 fulfilled the study criteria; 135 (50.6%) were men and the mean (SD) age was 65.4 (19.6) years . Aetiological agents were identified from 199 (75%) patients (one pathogen in 124 (46%), two in 53 (20%), and three or more in 22 (8%)): Streptococcus pneumoniae 129 (48%), influenza A virus 50 (19%), Chlamydia pneumoniae 35 (13%), Haemophilus influenzae 20 (7%), Mycoplasma pneumoniae 9 (3%), Legionella pneumophilia 9 (3%), other Chlamydia spp 7 (2%), Moraxella catarrhalis 5 (2%), Coxiella burnetii 2 (0.7%), others 8 (3%) . Atypical pathogens were less common in patients aged 75 years and over than in younger patients (16% v 27%; OR 0.5, 95% CI 0.3 to 0.9) . The 30 day mortality was 14.9% . Mortality risk could be stratified by the presence of four "core" adverse features . Three of 60 patients (5%) infected with an atypical pathogen died . CONCLUSION: S pneumoniae remains the most important pathogen to cover by initial antibiotic therapy in adults of all ages admitted to hospital with CAP . Atypical pathogens are more common in younger patients . They should also be covered in all patients with severe pneumonia and younger patients with non-severe infection.

Infect Immun, 2001 Apr, 69(4), 2748 - 52
Identification of the exported proteins of the oral opportunistic pathogen Actinobacillus actinomycetemcomitans by using alkaline phosphatase fusions; Ward J et al.; A phoA fusion library of Actinobacillus actinomycetemcomitans genomic DNA has been screened to identify genes encoding exported and secreted proteins . A total of 8,000 colonies were screened, and 80 positive colonies were detected . From these, 48 genes were identified with (i) more than half having homology to known or hypothetical Haemophilus influenzae genes, (ii) 14 having no ascribed function, and (iii) 4 having very limited or no homology to known genes . The proteins encoded by these genes may, by virtue of their presence on the cell surface, be novel virulence determinants.

Infect Immun, 2001 Apr, 69(4), 2684 - 91
Gallinacin-3, an inducible epithelial beta-defensin in the chicken; Zhao C et al.; Gallinacin-3 and gallopavin-1 (GPV-1) are newly characterized, epithelial beta-defensins of the chicken (Gallus gallus) and turkey (Meleagris gallopavo), respectively . In normal chickens, the expression of gallinacin-3 was especially prominent in the tongue, bursa of Fabricius, and trachea . It also occurred in other organs, including the skin, esophagus, air sacs, large intestine, and kidney . Tracheal expression of gallinacin-3 increased significantly after experimental infection of chickens with Haemophilus paragallinarum, whereas its expression in the tongue, esophagus, and bursa of Fabricius was unaffected . The precursor of gallinacin-3 contained a long C-terminal extension not present in the prepropeptide . By comparing the cDNA sequences of gallinacin-3 and GPV-1, we concluded that a 2-nucleotide insertion into the gallinacin-3 gene had induced a frameshift that read through the original stop codon and allowed the chicken propeptide to lengthen . The striking structural resemblance of the precursors of beta-defensins to those of crotamines (highly toxic peptides found in rattlesnake venom) supports their homology, even though defensins are specialized to kill microorganisms and crotamines are specialized to kill much larger prey.

Infect Immun, 2001 Apr, 69(4), 2636 - 42
CD8+ T cells have an essential role in pulmonary clearance of nontypeable Haemophilus influenzae following mucosal immunization; Foxwell AR et al.; A rodent respiratory experimental model has proved useful for investigating the immune mechanisms responsible for clearance of bacteria from the lungs . Immunohistochemical studies in immune and nonimmune rats have identified the cellular kinetics of response to bacterial pulmonary infection for CD8+, CD4+, and gammadelta+ T cells; B cells; and the expression of major histocompatibility complex class II (MHC-II) . During the course of bacterial clearance, there was no apparent proliferation or extravasation of lymphocytes, nor was there increased expression of MHC-II in nonimmune animals despite an influx of polymorphonuclear leukocytes, whereas in immunized animals there was an early influx of CD8+ and gammadelta+ T cells, followed by enhanced expression of the MHC-II marker, cellular infiltration by polymorphonuclear leukocytes, and finally an increased number of CD4+ T cells . Depletion of CD8+ T cells confirmed their vital contribution in the preprimed immune response to pulmonary infection by significantly decreasing the animals' ability to clear bacteria following challenge.

Infect Immun, 2001 Apr, 69(4), 2580 - 8
Mu-like Prophage in serogroup B Neisseria meningitidis coding for surface-exposed antigens; Masignani V et al.; Sequence analysis of the genome of Neisseria meningititdis serogroup B revealed the presence of an approximately 35-kb region inserted within a putative gene coding for an ABC-type transporter . The region contains 46 open reading frames, 29 of which are colinear and homologous to the genes of Escherichia coli Mu phage . Two prophages with similar organizations were also found in serogroup A meningococcus, and one was found in Haemophilus influenzae . Early and late phage functions are well preserved in this family of Mu-like prophages . Several regions of atypical nucleotide content were identified . These likely represent genes acquired by horizontal transfer . Three of the acquired genes are shown to code for surface-associated antigens, and the encoded proteins are able to induce bactericidal antibodies.

Infect Immun, 2001 Apr, 69(4), 2549 - 57
Haemophilus ducreyi associates with phagocytes, collagen, and fibrin and remains extracellular throughout infection of human volunteers; Bauer ME et al.; In a previous study, Haemophilus ducreyi was found in the pustule and dermis of samples obtained at the clinical end point in the human model of infection . To understand the kinetics of localization, we examined infected sites at 0, 24, and 48 h after inoculation and at the clinical end point . Immediately after inoculation, bacteria were found predominantly in the dermis but also in the epidermis . Few bacteria were detectable at 24 h; however, by 48 h, bacteria were readily seen in the pustule and dermis . H . ducreyi was associated with polymorphonuclear leukocytes and macrophages in the pustule and at its base, but was not associated with T cells, Langerhans' cells, or fibroblasts . H . ducreyi colocalized with collagen and fibrin but not laminin or fibronectin . Association with phagocytes, collagen, and fibrin was seen as early as 48 h and persisted at the pustular stage of disease . Optical sectioning by confocal microscopy and transmission electron microscopy both failed to demonstrate intracellular H . ducreyi . These data identify collagen and fibrin as potentially important targets of adherence in vivo and strongly suggest that H . ducreyi remains extracellular throughout infection and survives by resisting phagocytic killing in vivo.

Infect Immun, 2001 Apr, 69(4), 2378 - 82
Evaluation of De-O-acetylated meningococcal C polysaccharide-tetanus toxoid conjugate vaccine in infancy: reactogenicity, immunogenicity, immunologic priming, and bactericidal activity against O-acetylated and De-O-acetylated serogroup C strains; Richmond P et al.; The polysaccharide capsule of serogroup C Neisseria meningitidis (MenC) has been integral to vaccine development . Licensed MenC vaccines contain the O-acetylated (OAc+) form of polysaccharide . Some MenC strains have de-O-acetylated (OAc-) polysaccharide, which may affect antibody specificity and functional activity when used in a vaccine . We evaluated an OAc-MenC conjugate-tetanus toxoid conjugate (MCC-TT) vaccine given concomitantly with whole-cell diphtheria-tetanus-pertussis, Haemophilus influenzae type b, and oral polio immunization in 83 infants at 2, 3, and 4 months of age . Serum bactericidal activities (SBA) against OAc+ and OAc- MenC strains and OAc+ and OAc- polysaccharide-specific immunoglobulin G (IgG) levels were evaluated . MCC-TT vaccine was well tolerated . All infants produced SBA titers of > or = 8 after a single dose at 2 months of age . The SBA geometric mean titer for OAc+ strain C11 increased from 2.7 (95% confidence interval {CI} 2.2 to 3.2) to 320 (95% CI, 237 to 432), 773 (95% CI, 609 to 982), and 1,063 (95% CI, 856 to 1319) after one, two, and three doses of MCC-TT, respectively . OAc- IgG levels were twice as high as OAc+ IgG levels after the primary series of MCC-TT vaccine, and the SBA was significantly higher against the OAc- MenC strain . Antibody responses to booster vaccination with either OAc+ MenC polysaccharide vaccine (MACP) or a fourth dose of MCC-TT at 14 months of age provided evidence of immunologic memory . The acetylation status of the booster vaccine influenced the specificity of the response, with significantly higher OAc- IgG levels and SBA after MCC-TT vaccine compared to MACP vaccine but similar OAc+ antibody levels . MCC-TT vaccine is highly immunogenic and primes for immunologic memory against OAc+ and OAc- MenC strains in infancy.

Infect Immun, 2001 Apr, 69(4), 2353 - 63
Involvement of HxuC outer membrane protein in utilization of hemoglobin by Haemophilus influenzae; Cope LD et al.; Haemophilus influenzae can utilize different protein-bound forms of heme for growth in vitro . A previous study from this laboratory indicated that nontypeable Haemophilus influenzae (NTHI) strain N182 expressed three outer membrane proteins, designated HgbA, HgbB, and HgbC, that bound hemoglobin or hemoglobin-haptoglobin and were encoded by open reading frames (ORFs) that contained a CCAA nucleotide repeat . Testing of mutants expressing the HgbA, HgbB, and HgbC proteins individually revealed that expression of any one of these proteins was sufficient to allow wild-type growth with hemoglobin . In contrast, mutants that expressed only HgbA or HgbC grew significantly better with hemoglobin-haptoglobin than did a mutant expressing only HgbB . Construction of an isogenic hgbA hgbB hgbC mutant revealed that the absence of these three gene products did not affect the ability of NTHI N182 to utilize hemoglobin as a source of heme, although this mutant was severely impaired in its ability to utilize hemoglobin-haptoglobin . The introduction of a tonB mutation into this triple mutant eliminated its ability to utilize hemoglobin, indicating that the pathway for hemoglobin utilization in the absence of HgbA, HgbB, and HgbC involved a TonB-dependent process . Inactivation in this triple mutant of the hxuC gene, which encodes a predicted TonB-dependent outer membrane protein previously shown to be involved in the utilization of free heme, resulted in loss of the ability to utilize hemoglobin . The results of this study reinforce the redundant nature of the heme acquisition systems expressed by H . influenzae.

Arch Pediatr Adolesc Med, 2001 Mar, 155(3), 382 - 6
Effectiveness of a practice-based intervention to increase vaccination rates and reduce missed opportunities; Minkovitz CS et al.; BACKGROUND: Although provider feedback and recall/reminder systems have been shown to increase vaccination rates for children, little is known about the effectiveness of less intensive interventions . OBJECTIVE: To determine whether provider prompting at acute care visits in an urban hospital-based outpatient clinic can increase vaccination rates and decrease missed opportunities . DESIGN AND METHODS: Study participants, 3 years or younger, were identified from a managed care organization as receiving primary care at the clinic . Eligibility criteria included 1 or more visits to the clinic without regard to continuity of enrollment . Patients' vaccination records were generated at nursing triage and attached to the encounter sheet . Vaccination and visit data were abstracted from medical records, and comparisons were made between baseline (n = 521) and postintervention (n = 642) groups for up-to-date vaccination rates, missed opportunity rates, and mean numbers of visits . RESULTS: Up-to-date rates at the age of 24 months for 4 diphtheria and tetanus toxoids and pertussis, 3 polio, 1 measles-mumps-rubella, 3 hepatitis B, and 3 Haemophilus influenzae type b vaccines changed from 70% to 78% (P =.07) . Up-to-date rates increased significantly to 87% among the subset of children continuously enrolled in the managed care organization and the practice (P<.01) . Overall, mean numbers of visits were similar . Missed opportunity rates among children not up-to-date for 4 diphtheria and tetanus toxoids and pertussis, 3 polio, 1 measles-mumps-rubella, 3 hepatitis B, and 3 Haemophilus influenzae type b vaccines at the age of 24 months declined from 65% to 45% (P =.04) . Similar trends were noted at the age of 10 months . CONCLUSIONS: In the absence of increased funding, minor changes in standard operating procedures may improve vaccination delivery . Further improvements may require efforts to ensure continuity of provider and plan assignment.

Mol Microbiol, 2001 Feb, 39(4), 850 - 62
The variable P5 proteins of typeable and non-typeable Haemophilus influenzae target human CEACAM1; Hill DJ et al.; Haemophilus influenzae, a commensal of the human respiratory mucosa, is an important cause of localized and systemic infections . We have recently shown that numerous strains of capsulate (typeable) and acapsulate (non-typeable) H . influenzae target the carcinoembryonic antigen (CEA) family of cell adhesion molecules (CEACAMs) . Moreover, the ligands appeared to be antigenically variable and, when using viable typeable bacteria, their adhesive functions were inhibited by the presence of capsule . In this report, we show that the antigenically variable outer membrane protein, P5, expressed by typeable and non-typeable H . influenzae targets human CEACAM1 . Variants and mutants lacking the expression of P5 of all strains tested were unable to target purified soluble receptors . A non-typeable strain that did not interact with CEACAM1 was made adherent to both the soluble receptors and CEACAM1-transfected Chinese hamster ovary cells by transformation with the P5 gene derived from the adherent typeable strain Rd . However, several H . influenzae mutants lacking P5 expression continued to bind the cell-bound CEACAM1 receptors . These observations suggest that (i) CEACAM1 alone can support P5 interactions and (ii) some strains contain additional ligands with the property to target CEACAM1 but require the receptor in the cellular context . The identification of a common ligand in diverse strains of H . influenzae and the presence of multiple ligands for the same receptor suggests that targeting of members of the CEACAM family of receptors may be of primary significance in colonization and pathogenesis of H . influenzae strains.

Microbes Infect, 2001 Feb, 3(2), 161 - 6
Pulmonary collectins and innate host defense of the lung; LeVine AM et al.; Surfactant proteins A and D (SP-A and SP-D) are members of the collectin family of polypeptides expressed in the respiratory tract that bind bacterial, fungal and viral pathogens, enhancing their opsonization and killing by phagocytic cells . Clearance of bacterial pathogens including group B streptococci, Haemophilus influenza, Pseudomonas aeruginosa and viral pathogens, respiratory syncytial virus, adenovirus and influenza A virus, was deficient in SP-A(-/-) mice . SP-A deficiency was associated with enhanced inflammation and synthesis of proinflammatory cytokines . SP-D(-/-) mice cleared these bacteria as efficiently as wild-type mice; however, clearance of viral pathogens was deficient in SP-D(-/-) mice and associated with increased inflammation . SP-A and SP-D play critical and distinct roles in the regulation of alveolar macrophage function and inflammation, contributing to innate defense of the lung.

Bioorg Med Chem, 2001 Feb, 9(2), 465 - 75
Orally active cephalosporins . Part 3: synthesis, structure-activity relationships and oral absorption of novel C-3 heteroarylmethylthio cephalosporins; Yamamoto H et al.; A series of 7beta-{(Z)-2-(2-aminothiazol-4-yl)-2-hydroxyiminoacetamido}-3-(heteroarytmethylthio)cephalosporins was designed, synthesized and evaluated for antibacterial activity and oral absorption in rats . Antibacterial activity was markedly influenced by the structure of the heteroaromatic ring moiety . Oral absorption was influenced by the heteroaromatic ring moiety as well as by the arrangement of heteroatoms . Among these compounds, FK041 (2o), having a 4-pyrazolylmethylthio moiety, showed potent antibacterial activity against both gram-positive and gram-negative bacteria including Haemophilus influenzae . Further, it showed higher oral absorption than CFDN.

Proc Natl Acad Sci U S A, 2001 Mar 13, 98(6), 3460 - 5 Epub 2001 Mar 06.
Complete genomic sequence of Pasteurella multocida, Pm70; May BJ et al.; We present here the complete genome sequence of a common avian clone of Pasteurella multocida, Pm70 . The genome of Pm70 is a single circular chromosome 2,257,487 base pairs in length and contains 2,014 predicted coding regions, 6 ribosomal RNA operons, and 57 tRNAs . Genome-scale evolutionary analyses based on pairwise comparisons of 1,197 orthologous sequences between P . multocida, Haemophilus influenzae, and Escherichia coli suggest that P . multocida and H . influenzae diverged approximately 270 million years ago and the gamma subdivision of the proteobacteria radiated about 680 million years ago . Two previously undescribed open reading frames, accounting for approximately 1% of the genome, encode large proteins with homology to the virulence-associated filamentous hemagglutinin of Bordetella pertussis . Consistent with the critical role of iron in the survival of many microbial pathogens, in silico and whole-genome microarray analyses identified more than 50 Pm70 genes with a potential role in iron acquisition and metabolism . Overall, the complete genomic sequence and preliminary functional analyses provide a foundation for future research into the mechanisms of pathogenesis and host specificity of this important multispecies pathogen.

J Trop Pediatr, 2001 Feb, 47(1), 24 - 9
Safety and immunogenicity of combined or associated administration of PRP-T vaccine with diphtheria, tetanus and pertussis vaccine in Thai children; Lolekha S et al.; To assess whether the combination of a diphtheria, tetanus and pertussis vaccine with a Haemophilus influenzae type b conjugate vaccine (PRP-T) had any effect on immunogenicity or safety compared with separate administration of the vaccines, 158 infants were randomized to receive the vaccines either in association or as a combination at 2, 4, and 6 months of age . A total of 126 infants (59 associated, 67 combination) completed the three-dose regimen and were analysed for immunogenicity and safety . With respect to safety, there were no significant differences between the two groups . The combination vaccine was well tolerated with adverse reactions consisting mainly of transient pain, redness, induration and some low-grade fever . With respect to immunogenicity, response to PRP-T vaccine was good . Following just two doses, all infants achieved anti-PRP titers >0.15 microg/ml, regardless of whether the vaccine was given in combination or association . Following three doses, 98.5 per cent of infants in the combination group and 98.3 per cent in the association group had titers higher than 1.0 microg/ml.

Eur J Clin Microbiol Infect Dis, 2001 Jan, 20(1), 55 - 60
Antibiotic susceptibilities among recent clinical isolates of Haemophilus influenzae and Moraxella catarrhalis from fifteen countries; Bandak SI et al.; Between July 1998 and July 1999, 3,060 Haemophilus influenzae and 1,486 Moraxella catarrhalis strains were isolated in 31 centers in 15 countries in order to determine their antimicrobial susceptibilities and the presence of beta-lactamase production in Haemophilus influenzae . Overall 17.1% of the Haemophilus influenzae isolates were beta-lactamase positive, while more than 95% were susceptible to amoxicillin/clavulanate, cefaclor, loracarbef, cefuroxime, azithromycin and ciprofloxacin . Eleven (0.3%) isolates were beta-lactamase positive and ampicillin resistant and 7 (0.2%) isolates were ciprofloxacin resistant . The minimum inhibitory concentrations for 90% of the isolates tested were lowest for ciprofloxacin (0.03) and highest for cefprozil (8) against Moraxella catarrhalis.

ORL J Otorhinolaryngol Relat Spec, 2001 Mar-Apr, 63(2), 96 - 101
Viral co-infection does not reduce the efficacy of vaccination against non-typeable Haemophilus influenzae middle ear infection in a rat model; Moore R et al.; The mucosal vaccination of rodents with killed non-typeable Haemophilus influenzae (NTHi) has been previously shown to enhance live NTHi clearance following middle ear challenge . This study assessed the efficacy of mucosal anti-NTHi vaccination during a concomitant viral infection of the respiratory tract . Animals were mucosally immunised with killed NTHi by intra-Peyer's patch primary inoculation and lung (intratracheal) boost . At the time of both immunisations rats were also infected intra-nasally with Sendai virus . Concomitant Sendai virus infection did not influence the efficacy of anti-NTHi vaccination mediated clearance of NTHi from the middle ear . This would suggest that immunisation strategies to prevent bacterial middle ear infection would be effective despite the presence of concomitant viral agents .

Presse Med, 2001 Jan, Spec No 1, 21 - 2
{Haemophilus influenzae sensitivity to beta-lactamines}; Dabernat H; FACTS: Among the 280 strains studied, 106 were sensitive to beta-lactams, 92 were beta-lactamase producers, and 82 non-producers of beta-lactamase had reduced sensitivity to beta-lactams . QUESTION: The results of this study illustrate the complexity of characterizing the phenotypic resistance of H . influenzae and raises the question of which in vitro susceptibility tests best identify resistance.

Clin Diagn Lab Immunol, 2001 Mar, 8(2), 221 - 4
PCR-based detection, restriction endonuclease analysis, and transcription of tonB in Haemophilus influenzae and Haemophilus parainfluenzae isolates obtained from children undergoing tonsillectomy and adenoidectomy; Matar GM et al.; We developed and evaluated a PCR-based-restriction endonuclease analysis method to detect and analyze the tonB gene of Haemophilus influenzae and Haemophilus parainfluenzae from pediatric patients undergoing tonsillectomy and adenoidectomy . Multiple sites from the same patient, including the surface of adenoids and tonsils, as well as the core of tonsils, were cultured on chocolate agar and identified using standard procedures and the API NH Kit . A total of 55 H . influenzae isolates were recovered from different sites of 20 patients, and 32 H . parainfluenzae isolates were recovered from various sites of 12 patients . DNA was extracted from American Type Culture Collection strains and test isolates by the PureGene kit . Two primers, G1 (21-mer) and G2 (23-mer), were designed by us to amplify by PCR the tonB gene that consists of an 813-bp fragment . A nested PCR using primers T1 (23-mer) and T2 (24-mer) that flank an internal sequence to the gene of the order of 257 bp and restriction endonuclease digestion using XhoI and BglII were done to detect whether heterogeneity within the gene exists between the two species . Reverse transcription-PCR (RT-PCR) was finally done to detect transcription of the gene in both species . Our data have shown that the tonB gene was detected in both species . It is known to encode a virulent protein, TonB, in H . influenzae; however, demonstration of its presence in H . parainfluenzae is novel . Nested-PCR and restriction endonuclease analysis have shown that the tonB gene is apparently structurally the same in both species, with possible differences that may exist in certain H . parainfluenzae isolates . RT-PCR done on selected numbers of H . influenzae and H . parainfluenzae have shown that the tonB gene was transcribed in both species . This shows that the TonB protein, if expressed, may play a different role in the virulence in H . parainfluenzae since it is not needed for heme or heme complexes uptake as with H . influenzae.

J Infect Dis, 2001 Mar 15, 183(6), 880 - 6 Epub 2001 Feb 09.
Antibiotic treatment in acute Otitis Media promotes superinfection with resistant Streptococcus pneumoniae carried before initiation of treatment; Dagan R et al.; Antibiotic-resistant pneumococci are difficult to eradicate from middle ear fluid (MEF) and the nasopharynx (NP) . Bacteriologic eradication from the NP and MEF during acute otitis media (AOM) by 3 common antibiotic drugs was prospectively evaluated . In 19 (16%) of 119 MEF culture-positive patients, an organism susceptible to the treatment drug (Haemophilus influenzae, Streptococcus pneumoniae, or both) was isolated from the initial MEF, whereas resistant S . pneumoniae was present in the NP; in 9 (47%) patients, the initial resistant NP organism (identified by serotyping, resistance to the administered drug, and pulsed-field gel electrophoresis) replaced the susceptible MEF organism within only a few days after initiation of treatment . In regions where resistant pneumococci are prevalent, antibiotics may not only fail to eradicate the organisms, but they may often induce MEF superinfection with resistant pneumococci initially carried in the NP . This is an important mechanism by which, in recently treated patients, AOM infections often become refractory to treatment.

Biologicals, 2000 Dec, 28(4), 227 - 31
The determination of phosphorus in Haemophilus influenzae type b conjugate vaccines by inductively coupled plasma-atomic emission spectrometry; Swartz LA et al.; This study describes a method for the determination of phosphorus in lyophilized Haemophilus influenzae type b conjugate vaccines by inductively coupled plasma-atomic emission spectroscopy (ICP-AES) . The concentration of polysaccharide is directly related to the concentration of phosphorus as measured in the laboratory . Phosphorus is present in the polyribosyl-ribitol phosphate (PRP) group of the Haemophilus influenzae type b conjugate vaccine . The repeating unit of PRP is 3-B-D ribose{1-1}ribitol-5-phosphate . Phosphorus in the final container is measured in microg per dose . The amount of PRP is calculated from this and reported in microg per dose . The Haemophilus influenzae type b conjugate vaccine was analyzed for phosphorus content within the range of 1.34 to 2.02 microg phosphorus per ml . The relative difference of phosphorus concentrations determined by the ICP-AES method from the phosphorus concentrations determined by the traditional colorimetric molybdate method ranged from 2.2 to 10.6% . Phosphorus spike recovery for the vaccine ranged from 93 to 99% (1.93+/-0.13 microg P/ml) . The phosphorus determination of NIST SRM 3139 phosphorus spectrometric solution differed by 3.0% from the certified phosphorus value (10.00 mg P/ml).

J Med Microbiol, 2001 Mar, 50(3), 277 - 83
Use of automated riboprinter and pulsed-field gel electrophoresis for epidemiological studies of invasive Haemophilus influenzae in Taiwan; Wang CC et al.; A total of 87 invasive isolates of Haemophilus influenzae isolated throughout Taiwan from 1994 to 1998 was collected; 57 were from children <14 years old . In all, 60.9% of isolates were resistant to ampicillin and produced beta-lactamase . Ribotyping revealed six different profiles in 55 isolates of type b, nine profiles in 10 isolates of non-type b and 12 profiles in 22 isolates of non-typable H . influenzae . Among isolates from 35 cases of meningitis, 30 (86%) were in ribogroups 1, 2 and 3 with >90% genetic similarity . Compared with all the other ribogroups, ribogroups 1, 2 and 3, which encompassed all H . influenzae type b, were significantly more prevalent as a cause of meningitis in children <14 years old . Further subtyping of the predominant ribogroup by pulsed-field gel electrophoresis (PFGE) identified differences of 0-6 bands among these isolates of ribogroup 1, which indicated distant relatedness . Automated ribotyping was found to be a useful method and was less time-consuming for molecular epidemiology studies of H . influenzae . PFGE is suggested as an addition to ribotyping to improve discrimination if H . influenzae type b is involved . Differentiating ribogroups between type b and non-type b H . influenzae by genotyping may help to understand the molecular characteristics of outbreaks, endemicity and value of vaccination . According to the results of ribotyping and PFGE, it seems possible that spread of invasive H . influenzae type b had occurred and ribotyping confirmed that there was no clonal spread of non-type b H . influenzae in Taiwan.

Arch Pediatr, 2001 Feb, 8(2), 205 - 10
{The child with recurrent infections: which screening for immunoe deficiency?}; Siegrist CA; A child with recurrent infections represents a challenge to the pediatrician who must identify, among a large number of repeatedly infected but nevertheless healthy children whose parents need to be reassured, the rare cases of potentially severe immune deficiency . This can be most successfully achieved through the measurement of IgA, IgG, and antibody titers to vaccine (tetanus, diphtheria, Haemophilus influenzae B) and exposure (pneumococcus) antigens . The presence of normal antibody responses makes it possible to rule out underlying immune deficiency in a sensitive and specific manner . Conversely, abnormally weak antibody responses identify the children who have to be referred without delay for further investigation of a potential immune defect . This article indicates for which pediatric patients an immunodeficiency screening should be considered, and how to analyze its results.

J Mol Evol, 2001 Feb, 52(2), 164 - 70
Correlation between Shine--Dalgarno sequence conservation and codon usage of bacterial genes; Sakai H et al.; In this study, we analyzed the correlation between codon usage bias and Shine--Dalgarno (SD) sequence conservation, using complete genome sequences of nine prokaryotes . For codon usage bias, we adopted the codon adaptation index (CAI), which is based on the codon usage preference of genes encoding ribosomal proteins, elongation factors, heat shock proteins, outer membrane proteins, and RNA polymerase subunit proteins . To compute SD sequence conservation, we used SD motif sequences predicted by Tompa and systematically aligned them with 5'UTR sequences . We found that there exists a clear correlation between the CAI values and SD sequence conservation in the genomes of Escherichia coli, Bacillus subtilis, Haemophilus influenzae, Archaeoglobus fulgidus, Methanobacterium thermoautotrophicum, and Methanococcus jannaschii, and no relationship is found in M . genitalium, M . pneumoniae, and Synechocystis . That is, genes with higher CAI values tend to have more conserved SD sequences than do genes with lower CAI values in these organisms . Some organisms, such as M . thermoautotrophicum, do not clearly show the correlation . The biological significance of these results is discussed in the context of the translation initiation process and translation efficiency.

Fresenius J Anal Chem, 2000 Dec, 368(8), 739 - 58
Carbohydrate analysis by high-performance anion-exchange chromatography with pulsed amperometric detection: the potential is still growing; Cataldi TR et al.; This article reviews recent advances of carbohydrate analysis by high-performance anion-exchange chromatography with pulsed amperometric detection . Starting from the paper of Dennis C . Johnson {1} in which the great analytical promise of such a technique was anticipated, a multitude of exciting new research possibilities have recently emerged . The great attractiveness of high-performance anion-exchange chromatography is largely due to its compatibility with such a sensitive, selective and reliable detection method as pulsed amperometry . This very good match between liquid chromatography and electrochemical detection has allowed the determination of carbohydrates in a variety of complex matrices, for instance, foods, beverages, diary and biotechnological products, vegetal tissues, and also in the area of clinical diagnostics . For this reason, the introduction of HPAEC-PAD into regulated methods is becoming increasingly accepted . A comprehensive collection of applications to carbohydrates and samples of interest is given, with special focus on the separation of closely related sugar compounds using dilute alkaline eluents . Advances in pulsed potential waveforms are also discussed, and a comparison with other liquid chromatographic methods addressed . 2-keto-3-deoxy-D-glycero-D-galactonononic acid; KDO, 2-keto-3-deoxyoctulosonic acid; FOS, fructooligosaccharides; GF5, GF6, and GF7, oligofructans: Hib, Haemophilus influenzae type b; FAB, fast atom bombardment; ESI, electrospray ionization; MALDI-TOF, matrix assisted laser desorption ionization-time of flight.

Acta Paediatr, 2001 Jan, 90(1), 99 - 101
Impaired IgG responses in a child with homozygous C2 deficiency and recurrent pneumococcal septicaemia; Attwood JT et al.; Homozygous deficiency of the second component of complement (C2) is the most common inherited deficiency of complement . Although C2 deficiency has been detected in asymptomatic individuals, patients usually present with either autoimmune disease or recurrent pyogenic infection, particularly due to encapsulated bacteria such as Streptococcus pneumoniae, Haemophilus influenzae type b and Neisseria meningitidis . Interestingly, infection is the most common mode of presentation of C2 deficiency in young children (1) . An association between C2 deficiency and IgG subclass deficiency has also been previously described . We now report a female child with C2 deficiency that presented at the age of 3 mo with recurrent pneumococcal septicaemia . Although IgG subclass levels were normal, specific IgG responses to vaccination against S . pneumoniae and H . influenzae were significantly impaired.

Can J Vet Res, 2001 Jan, 65(1), 7 - 14
Seasonal incidence and antibiotic susceptibility patterns of Pasteurellaceae isolated from American bison (Bison bison); Dyer NW et al.; Ninety pharyngeal tonsils were collected from 2-year-old American bison (Bison bison) bulls and sampled for members of the Pasteurellaceae family . Particular attention was paid to seasonal incidence and antimicrobial resistance in serotypes and biovariants . Multiple strains of Pasteurella haemolytica (39%), P . trehalosi (68%), P . multocida (34%) and Haemophilus somnus (13%) were cultured from 86 out of the 90 (96%) tonsil samples . Pasteurella trehalosi was the most common and evenly distributed of the organisms recovered . Pasteurella haemolytica was found in fewer numbers than P . trehalosi, but showed an increase in number of isolates recovered with each sampling period . Pasteurella multocida, both A and D capsular types, was recovered from all sampling periods . No serotype pattern was observed in any of the animal groups sampled . One hundred twenty-seven of 147 (86%) of the isolates were resistant to at least 1 antibiotic, 95/147 (65%) to at least 2 different antibiotics, and 16/147 (11%) to at least 3 antibiotics . The most common resistance pattern observed was to neomycin and spectinomycin (73/147) (49%).

Am J Trop Med Hyg, 2000 Apr, 62(4), 485 - 90
Randomized trial of the immunogenicity of fractional dose regimens of PRP-T Haemophilus influenzae type b conjugate vaccine; Fernandez J et al.; To assess the immunogenicity of more economical regimens of Haemophilus influenzae type b (Hib) conjugate vaccine, a randomized trial of fractional doses of polyribosylribitol phosphate-tetanus toxoid (PRP-T) Hib vaccine was undertaken in the Dominican Republic . Six hundred children were assigned to one of six regimens with PRP-T vaccine: full-dose, half-dose, and one-third-dose of Hib vaccine given separately or combined with diphtheria, tetanus, and pertussis (DTP) vaccine at ages 2, 4, and 6 months . Regimens that elicited antibody levels > 1.0 microg/mL in >70% of children and < or = 0.15 microg/mL in > 90% of children were considered acceptable . At 1 month post Dose 3, all regimens met the criteria for acceptable response . Among those who received Hib as a separate injection, geometric mean concentrations of anti-PRP bodies (GMCs) at age 1 month post Dose 3 were 11.2, 11.9, and 16.3 in the full, half, and one-third dose groups, respectively . Among those who received Hib and DTP combined, the GMCs were 6.4, 5.2, and 5.7 in the full-, half-, and one-third-dose groups respectively.

Med Arh, 2000, 54(5-6), 317 - 8
{Clinical assessment of occult infections in children}; Sporisevic L et al.; Children's occult infections are characterised presenting pathogenic bacteries in blood of children in age 3 to 36 months, but they are good general aspect and orderly immunologic status and they don't have signs of focal infection . Manifestation of occult infections determined: age of child, increasing bodies temperature, testsphysical observance and clinical-biochemistry tests . Prevalence of manifestation occult infections is 3-8%, but they manifest ni a form occult bacteremia, occult pneumonia nad occult urinary infection . Methodic, systematic admission and adequate clinical-biochemical monitoring, we minimise sequeles of occult infections . Risk of serious sequeles at occult infections is importantly decreasing by epidemiological changes that it rises by using vaccination against Haemophilus influenzae and Streptococcus pneumoniae is leading ethiological source . Many contraversal opinions are presented in glance of therapeutic strategy at children's occult infection . Future of solutions at many hesitations ni context diagnosis and therapy of occult infections is established in using recent detectional tests /pneumococcus PCR, plasmas tumor reaction, interleukin la/ and preventive intervetions activities /conjugated pneumococcus vaccination/.

Acta Crystallogr D Biol Crystallogr, 2001 Mar, 57(Pt 3), 418 - 20
Crystallization and preliminary X-ray analysis of the recombinant dihaem cytochrome c (NapB) from Haemophilus influenzae; Brige A et al.; The napB gene of the pathogenic bacterium Haemophilus influenzae encodes a dihaem cytochrome c, the small subunit of a heterodimeric periplasmic nitrate reductase (Nap) . Recombinant NapB was overproduced in Escherichia coli, purified to near-homogeneity and crystallized using the hanging-drop method . Thin quadrilateral plates were grown under various conditions but proved to be unsuitable for X-ray analysis . However, a single crystal was grown using 1.75 M ammonium sulfate in 0.1 M sodium acetate pH 5.5, from which a native data set could be collected to 1.8 A resolution using synchrotron radiation . Using the same conditions, further crystals were obtained by microseeding . The space group was determined to be P42(1)2, with unit-cell parameters a = 77.55, b = 77.55, c = 28.64 A and an unusually low solvent content of 16.5%, assuming there to be one molecule of NapB in the asymmetric unit . Analysis of the dissolved crystals indicated that partial proteolysis of the protein had occurred . Taking the molecular mass of the crystallized form ( approximately 8500 Da) into account, the solvent content was estimated to be 53%, with a V(M) value of 2.64 A(3) Da(-1).

Ann Otol Rhinol Laryngol, 2001 Feb, 110(2), 148 - 51
Correlation between microbiology and previous sinus surgery in patients with chronic maxillary sinusitis; Brook I et al.; Aspirates of 108 chronically inflamed maxillary sinuses were processed for aerobic and anaerobic bacteria . There were 295 bacterial isolates: 109 aerobic and facultative, and 186 anaerobic . The predominant aerobic isolates were Staphylococcus aureus (17 isolates), alpha-hemolytic streptococci (14), Pseudomonas aeruginosa (12), Moraxella catarrhalis (10), and Haemophilus spp (8) . The predominant anaerobes were Peptostreptococcus spp (61), Prevotella spp (45), Fusobacterium spp (15), and Propionibacterium acnes (14) . Analysis of the medical histories revealed a correlation only between the microbial results and previous sinus surgery . Pseudomonas aeruginosa and gram-negative aerobic bacilli (GNAB) were more often isolated in patients who had surgery (9 of 33 patients had P aeruginosa and 17 had GNAB) than in patients who did not have surgery (3 of 75 had P aeruginosa and 7 had GNAB; p < .001) . Anaerobes were isolated more often in patients who did not have surgery (69 of 75 patients) than in those who had previous surgery (21 of 33 patients; p < .001) . These findings illustrate the unique microbiological features of chronic maxillary sinusitis that persist after sinus surgery.

Ned Tijdschr Geneeskd, 2001 Feb 3, 145(5), 211 - 4
{CBO-guideline 'Bacterial meningitis'}; Roord JJ et al.; Neisseria meningitidis and Streptococcus pneumoniae are the most frequent causes of bacterial meningitis . The incidence of Haemophilus meningitis in the Netherlands is low due to successful Haemophilus influenzae type b vaccination . This implies that there is no need to take account into this microorganism in using initial empiric antimicrobial therapy for bacterial meningitis . Vomiting (especially children), headache, fever, and a stiff neck characterize acute bacterial meningitis . However, even without these signs a patient may still have acute bacterial meningitis . The characteristics in neonates are less specific . An emergency lumbar puncture should be performed in all patients with meningeal irritation or other signs of bacterial meningitis . Examination of the CSF is not indicated for convulsive children (between the ages of 6 months and 6 years) who do not exhibit other clinical signs . In patients who respond adequately to the treatment, it is not necessary to examine the CSF again . Papilloedema or focal neurological symptoms contraindicate a lumbar puncture in patients with bacterial meningitis, until CT results justify that it can be performed safely . Antibiotic treatment should not be delayed until after the CT . General practitioners should treat their patients with suspected meningococcus infection by admitting them to the hospital without first injecting antibiotics . In the Netherlands, patients with suspected pneumococcus meningitis may still be treated with benzylpenicillin . Patients with bacterial meningitis have no fluid restrictions; only in case of the syndrome of inadequate secretion of antidiuretic hormone is fluid reduction indicated . The physician is responsible for prescribing prophylaxis to family members . The Regional Health Services organize chemoprophylaxis for classmates . The latter is only indicated if at least 2 related cases occur in one month.

Rinsho Shinkeigaku, 2000 Aug, 40(8), 836 - 9
{A case of acute motor sensory axonal polyneuropathy after Haemophilus influenzae infection}; Oda M et al.; A 47-year-old woman developed consciousness disturbance, and experienced hallucinations while traveling abroad, and then went into critical condition . She was placed in the critical care unit, and had flaccid tetraparesis requiring mechanical ventilation . Haemophilus influenzae was cultured from the sputum . The level of protein of the cerebrospinal fluid was elevated to 114 mg/dl, nerve conduction study showed findings of pure axonal damage, and the sural nerve biopsy revealed severe axonal degeneration . She improved gradually by plasma exchange . The diagnosis of acute motor sensory axonal polyneuropathy (AMSAN) based on autoimmune mechanism was made . We speculate that H . influenzae infection may have elicited AMSAN in this case.

Kansenshogaku Zasshi, 2001 Jan, 75(1), 42 - 7
{Comparison of community-acquired pneumonia in relation to influenza A and RS virus infections}; Kobashi Y et al.; We experienced 142 cases with community-acquired pneumonia between April 1998 and March 2000 . By measuring the titers of respiratory viruses for these cases, we were able to identify acute phase infections of influenza A virus in 10 cases and RS virus in 6 cases and determined that there was an increase in community-acquired pneumonia during both winter seasons . Thereafter we compared the clinical features of community-acquired pneumonia with regard to these two types of virus infection by dividing the patients into two groups, both of which frequently included in the elderly . In the influenza virus group, such general symptoms as high fever, headache and general fatigue were dominant . Common bacteria were isolated in nine cases with mixed infection; four of them with Streptococcus pneumoniae . In the RS virus group, there were fewer general symptoms and common bacteria were isolated in four cases with mixed infection; three with Haemophilus influenzae . The severity of the illness was greater in the Influenza virus group; i.e.) three cases required mechanical ventilation and two of these three cases died . In the RS virus group, on the other hand, the prognosis was good because no mechanical ventilation was required and there were no deaths . Influenza vaccination is especially important for the elderly, because the epidemiology of the influenza virus groups showed none had a history of influenza vaccination in this study.

Epidemiol Infect, 2000 Dec, 125(3), 583 - 91
Dynamics of natural immunity caused by subclinical infections, case study on Haemophilus influenzae type b (Hib); Leino T et al.; Natural immunity to Haemophilus influenzae type b (Hib) is based primarily on antibodies that are thought to develop in response to subclinical infections . Wide use of conjugated Hib vaccines could lead to decreases in circulating Hib bacteria, thereby diminishing antibody levels in the unvaccinated . We applied a statistical model to estimate the duration of natural immunity to Hib under different forces of infection . Prior to the introduction of conjugated Hib vaccines, new Hib infections were estimated to occur once in 4 years and the antibody concentration to stabilize at a level around 1 microg/ml . In the absence of new stimuli, i.e . infection, 57% of the unvaccinated population would become susceptible to invasive disease (antibody levels < 0.15 microg/ml) in 10 years . Due to an interaction between the force of infection and the duration of immunity, in some situations numbers of invasive infections could increase in unvaccinated cohorts . This theoretical scenario has yet to be observed in practice.

Epidemiol Infect, 2000 Dec, 125(3), 549 - 54
Population-based surveillance for bacterial meningitis in the Dominican Republic: implications for control by vaccination; Gomez E et al.; Quantifying the local burden of disease is an important step towards the introduction of new vaccines, such as Haemophilus influenzae type b (Hib) conjugate vaccine . We adapted a generic protocol developed by the World Health Organization for population-based surveillance of bacterial meningitis . All hospitals that admit paediatric patients with meningitis in the National District, Dominican Republic were included in the system and standard laboratory methods were used . The system identified 111 cases of confirmed bacterial meningitis . Hib was the leading cause of bacterial meningitis, followed by group B streptococcus, S . pneumoniae, and N . meningitidis . Unlike hospital-based case series, this population-based system was able to calculate incidence rates . The incidence of Hib meningitis was 13 cases per 100,000 children < 5 years old . The data from this study were used by the Ministry of Health to support the introduction of routine Hib vaccination and will be used to monitor its effectiveness.

J Environ Pathol Toxicol Oncol, 2001, 20(1), 27 - 32
Molecular basis of UV-sensitive mutant strain MBH3 of Haemophilus influenzae Rd: identification of mutation in the uvrA gene; Balsara RD et al.; We have previously reported on cloning a DNA repair gene designated as uvr3 by virtue of its ability to phenotypically complement the UV sensitivity of mutant strain MBH3 . Subsequently, we identified the uvr3 gene to be the uvrA gene (gene identification number HI0249) of Haemophilus influenzae Rd . The uvrA gene is a component of the UvrABC excision repair pathway . We studied molecular basis of the UV sensitivity of the MBH3 strain and identified a G-->A transition at nucleotide position 2700 of the uvrA gene, altering the Trp-900 codon (TGG) to a nonsense codon (TGA) . Thus, the UvrA protein produced in the mutant strain MBH3 is likely to be truncated and unable to carry out the UV-induced DNA repair, thereby rendering the strain UV sensitive.

Dev Biol (Basel), 2000, 103, 35 - 47
Physicochemical characterisation of the oligosaccharide component of vaccines; Ravenscroft N et al.; Glycoconjugate vaccines are being developed against Haemophilus influenzae (Hib) and meningococcal Type A and C micro-organisms; they consist of oligosaccharides of intermediate chain length conjugated to the carrier protein CRM (a non-toxic diphtheria toxin mutant) . The oligosaccharides can be quantified using specific composition analyses and their structure and identity (and pattern of acetylation) evaluated by use of NMR spectroscopy . The average molecular-size (degree of polymerisation) can be determined using colorimetric assays, qualified by analysis of authentic standards . The molecular-size distribution of these anionic oligosaccharides can be achieved using ion exchange chromatography or application of the rapid and sensitive analytical HPAEC-PAD system (high performance anion-exchange chromatography with pulsed amperometric detection) . Preparative ion exchange chromatography permits the isolation of purified oligomers, which can be well-characterised using the methods described above . Molecular size can be confirmed by use of mass spectrometry . These vaccines are semi-synthetic products and therefore their preparation involves several steps of chemical reaction, the detailed physicochemical characterisation of the oligosaccharide-components permits the consistent production of these well-defined glycoconjugate vaccines.

Dev Biol (Basel), 2000, 103, 201 - 4
Properties of recombinant HtrA: an otitis media vaccine candidate antigen from non-typeable Haemophilus influenzae; Cates GA et al.; Non-encapsulated or non-typable Haemophilus influenzae (NTHi) is a major cause of middle ear infections in young children . HtrA has been identified as a vaccine candidate antigen from NTHi; therefore physicochemical characterization of this antigen is important for vaccine development . Recombinant NTHi HtrA has been expressed in E . coli and shown to have serine protease activity . Several mutant, recombinant HtrA proteins were expressed and purified to obtain suitable vaccine antigens lacking protease activity . Two mutants with alterations at the putative active site His91 and Ser197, designated H91A and S197A were examined by circular dichroic spectropolarimetry (CD) to evaluate secondary structure . The S197A mutant had a more random secondary structure compared to wild-type rHtrA or H91A . It is likely that improper folding of S197A accounts for its lack of immunoprotective properties in a chinchilla model of otitis media.

Monaldi Arch Chest Dis, 2000 Oct, 55(5), 415 - 9
Acute exacerbations in chronic obstructive pulmonary disease (COPD)--microbial patterns and risk factors; Schafer H et al.; Around 25% of patients with stable chronic obstructive pulmonary disease (COPD) show some evidence of tracheobronchial colonization . It is, however, probable that the vast majority of patients become colonized at some time during the course of the disease . A variety of factors including current smoking and viral infections predispose to bacterial colonization and subsequently acute exacerbations . In fact, infectious aetiologies account for around 50-75% of acute COPD exacerbations . Bacterial pathogens are present in around 50% of patients, Haemophilus influenzae and Streptococcus pneumoniae being the most frequently encountered pathogens . Pseudomonas aeruginosa and probably also Gram-negative enteric bacilli are more frequently found in patients with more severe airflow limitation . Viral infections are present in around 10-20%, with Influenzavirus representing the most frequent viral pathogen . Only recently, evidence for infection by Chlamydia pneumoniae has been found in 5-20% of patients . Predictors of distinct aetiologies have only infrequently been studied so far . Thus, it currently remains difficult to make adequate predictions on clinical grounds in the individual patient . Nevertheless, we would advocate to tentatively stratify the initial antimicrobial treatment according to the severity of the acute exacerbation episode and the presence of individual risk factors . The validation of such an approach may result in significant progress in our understanding of the role of infection and infectious agents in different subgroups of patients with acute exacerbations.

Onderstepoort J Vet Res, 2000 Dec, 67(4), 301 - 5
Pasteurella gallinarum: Zimbabwean experience of a versatile pathogen; Mohan K et al.; Pasteurella gallinarum-related outbreaks in chickens and African guinea fowls are described . Four outbreaks were recorded in chickens and one in guinea fowls . Periorbital swelling and keratoconjunctivitis were the consistently present clinical signs in all the diseased birds . In several, swollen hocks and wattles were also discerened . Birds which succumbed to the infection showed petechiation in the internal organs and evidence of airsacculitis . Pasteurella gallinarum was isolated from the lesions and also from conjunctival swabs of the apparently healthy in-contact birds . There was no evidence of concurrent infection with Haemophilus, Mycoplasma or Chlamydia . Quinolone therapy when resorted to on one of the farms resolved the clinical signs . Phenotypes of 28 isolates were studied . The results compared well with the Pasteurella gallinarum isolates reported earlier from elsewhere . It was also found that results of xylose fermentation and ONPG test appear to be a variable character . There is no earlier report of P . gallinarum infection in guinea fowls.

Eur J Clin Microbiol Infect Dis, 2000 Dec, 19(12), 926 - 31
Seven-year study of bacteraemic pneumonia in a single institution; Bishara J et al.; In order to enhance current knowledge of nosocomial and community-acquired bacteraemic pneumonia in a single tertiary hospital in Israel, a 7-year study was conducted . Using a computerised database, all patients who had bacteraemic pneumonia from March 1988 to August 1995 were studied . During the study period, pneumonia was the source of bacteraemia in 319 of 4,548 (7%) episodes, occurring in 295 patients; 211 (66%) episodes were community-acquired and 108 (34%) were nosocomial . The microoroganisms isolated most frequently from patients with community-acquired bacteraemic pneumonia were Streptococcus pneumoniae (46%), Staphylococcus aureus (10%) and Haemophilus influenzae (8%); while Pseudomonas spp . (17%), Klebsiella spp . (11%) and Staphylococcus aureus (10%) were isolated most often from the patients with nosocomial bacteraemic pneumonia . The median age of patients was 68 years (range, 0.003-100) . The overall mortality was 34% . No significant difference was found between the mortality rates of patients with community-acquired (31%) and nosocomial (40%) bacteraemic pneumonia (P=0.1) . Multivariate analysis showed that hypothermia, respiratory failure, impaired consciousness, tracheal intubation, Staphylococcus aureus aetiology, septic shock, inappropriate empiric antibiotic treatment and age significantly increased mortality.

Otolaryngol Pol, 2000, 54(5), 541 - 6
{Bacteriologic evaluation of middle ear fluid during the course of secretory otitis media in children}; Wezyk MT et al.; Fluid collected from tympanic cavity during operation from 43 children with O.M.S . was subjected to bacteriological examination . After the collection, each sample was immediately applied to Bactec Peds Plus/F liquid medium from Becton Dickinson . The number of samples, where each genus and species of bacteria were found, was determined, as well as the number of samples where every two genus coexisted; also the pH genes, in which each genus was found were calculated . Obtained results showed that 22 species of bacteria, belonging to 6 genus lived in the samples . The most commonly occurring genus were Staphylococcus, Streptococcus and Haemophilus; the most rare genus were Moraxella and Bacillus . The most common species were: Haemophilus influenzae (18.6% of samples), Staphylococcus aureus (14.0%), Staphylococcus epidermidis (11.6%), Staphylococcus warneri (9.3%) and Streptococcus oralis (7.0%) . Streptococcus coexisted most frequently with Staphylococcus and Haemophilus; Staphylococcus--with Haemophilus and Bacillus; Haemophilus--with Streptococcus and Staphylococcus . The pH ranges for the three most often found genus were: for Staphylococcus--7.7-9, for Streptococcus--7.7-9.3 and for Haemophilus--8.2-8.8.

Ann Otol Rhinol Laryngol, 2001 Jan, 110(1), 87 - 90
Microbiology of serous otitis media in children: correlation with age and length of effusion; Brook I et al.; The purpose of this study was to correlate the microbiology of serous otitis media in children with the duration of the condition and the patient's age . Aspirates of serous ear fluids from 114 children were examined for aerobic and anaerobic bacteria . Bacterial growth was noted in 47 patients (41%) . Aerobic organisms only were recovered in 27 aspirates (57% of the culture-positive aspirates); anaerobic bacteria only in 7 (15%); and mixed aerobic and anaerobic bacteria in 13 (28%) . A total of 83 bacterial isolates were recovered, accounting for 1.8 isolates per specimen (1.2 aerobes and 0.6 anaerobe) . There were a total of 57 aerobic isolates, including Haemophilus influenzae (15 isolates), Streptococcus pneumoniae (13), and Staphylococcus sp (12) . Twenty-six anaerobes were recovered, including anaerobic gram-positive cocci (10), Prevotella spp (8), and Propionibacterium acnes (4) . The rate of positive cultures (20 of 36; 56%) was higher in patients younger than 2 years of age than in those older than 2 years of age (27 of 78; 35%) . Streptococcus pneumoniae and H influenzae were more often isolated in children younger than 2 years of age and those with effusion for 3 to 5 months, whereas anaerobes were recovered more often in those older than 2 years of age and those with effusion for 6 to 13 months . These data illustrate the effects of the length of effusion and age on the recovery of aerobic and anaerobic bacteria in serous otitis media.

Med J Malaysia, 2000 Sep, 55(3), 304 - 7
Intravenous followed by oral ofloxacin in the treatment of community acquired lower respiratory tract infections in adults requiring hospitalisation; Liam CK et al.; Forty patients were treated with ofloxacin for community acquired lower respiratory tract infections . Eighteen pathogens were isolated in sputum; Streptococcus pneumoniae (4) and Haemophilus influenzae (4) were the most common, followed by Klebsiella pneumoniae (3), Klebsiella spp . (2), Staphylococcus anreus (2), Pseudomonas spp . (2), and Pseudomonas aeruginosa (1) . Ofloxacin 200 mg every 12 hours was administered for an average of 3.7 days intravenously followed by 5.4 days orally . Response to therapy was judged to be cure in 38 (95%; 95% C.I., 85%-95%) patients, failure in one (2.5%) and "indeterminate" in one (2.5%).

An Otorrinolaringol Ibero Am, 2000, 27(6), 541 - 9
{Nasopharyngeal bacterial flora and secretory otitis in adults}; Lacosta Nicolas JL et al.; The AA . have realized a prospective study of the links between the bacterial flora of the nasopharynx and the secretory otitis media in grown-up people . For achieving this purpose nasopharyngeal smears of rhinopharyngeal samples belonging to 85 otitic patients and other 85 healthy adults were cultivated . Statistical analysis showed that the otitis cases presented with 63.6% of microorganisms potentially pathogenic, being the 17.6% the percentage among healthy individuals (p < 0.001) . Microorganisms more frequently encountered were: Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis and Staphylococcus aureus . After the medical treatment those patients recovered showed an increase of saprophyte flora from early 36.4% till 86% (p < 0.001) . It could be appreciate a seasonal influence in the debut of this malady specially in winter.

Ann Pharmacother, 2001 Jan, 35(1), 36 - 40
Intramuscular ceftriaxone in the treatment of childhood meningitis due to Haemophilus influenzae type F; Ratka A et al.; OBJECTIVE: To describe a case of meningitis caused by Haemophilus influenzae type f (Hif) in a child . CASE SUMMARY: A 2.5-year-old white girl (18 kg) was hospitalized because of acute ataxia . The cerebrospinal fluid culture grew H . influenzae, which was later identified as type f . Therapy was limited by the inability to gain intravenous access . Treatment was initiated with dexamethasone 8 mg (0.44 mg/kg) intramuscularly, one dose on the day prior to initiation of ceftriaxone therapy, and intramuscular ceftriaxone 2 g (111 mg/kg/dose) once a day . After the first day, dexamethasone was administered at 3 mg (0.17 mg/kg/d) orally four times per day for four days . Within two days, the patient became afebrile and improved significantly . The remaining treatments were given during daily hospital visits on an outpatient basis . No complications occurred during the follow-up visits . DISCUSSION: The clinical presentation and therapeutic management of Hif meningitis is similar to that of H . influenzae type b (Hib) meningitis . Factors that may predispose a child to infections caused by Hif include upper respiratory tract infections, day care attendance, Down syndrome, and immunodeficiency . Hif meningitis usually is treated with a third-generation cephalosporin (frequently ceftriaxone) . Although most often administered intravenously, intramuscular ceftriaxone can provide a satisfactory clinical outcome in a child with adequate peripheral perfusion but limited intravenous access . The majority of reported cases of Hif meningitis resolve with appropriate antibiotic therapy; however, long-term neurologic sequelae occasionally occur . CONCLUSIONS: Hif occasionally causes pediatric meningitis . In a patient with good perfusion and difficult intravenous access, daily intramuscular administration of ceftriaxone can be an effective treatment option . In this case, Hif meningitis occurred abruptly and resolved within 48 hours of initiation of ceftriaxone and dexamethasone without long-term sequelae . The risks of giving dexamethasone appear to be minimal, although efficacy for preventing Hif complications remains to be proven.

Avian Dis, 2000 Oct-Dec, 44(4), 869 - 73
Relationship between antigen release and antibody response of infectious coryza water-in-oil-in-water emulsion vaccines; Fukanoki S et al.; The relationship between the antigen release from formulations in vitro and the antibody response after administration of water-in-oil-in-water (W/O/W) emulsion vaccines containing Haemophilus paragallinarum (Hpg) was studied in chickens . Increases of sorbitan sesquioleate volume in the formulation led to slower antigen release and tended to induce higher hemagglutination-inhibition (HI) antibody titers . In addition, the vaccines prepared with internal aqueous phase:oil phase:external aqueous phase (A:O:A) ratios of 3:4:3 and 3:3:4 also showed slower release of antigen and higher HI antibody titer compared with those of an A:O:A ratio of 3:2:5 . Vaccines prepared with polyoxyethylene (POE)(10) hydrogenated castor oil or POE(40) hydrogenated castor oil instead of sorbitan sesquioleate showed higher release and lower antibody HI titers . As a result, HI antibody titers at 6 wk after vaccination were inversely related to antigen release, as determined by the release test . The correlation coefficient was 0.942 . In infectious coryza W/O/W emulsion vaccines, the slow release of antigen from the formulation induced and maintained high HI antibody titers of Hpg.

Aust Vet J, 2000 Nov, 78(11), 759 - 62
Characterisation of isolates of Haemophilus paragallinarum from Indonesia; Poernomo S et al.; OBJECTIVE: To characterise 18 isolates of Haemophilus paragallinarum isolated from chickens in Indonesia . PROCEDURE: The isolates were identified to species level by traditional phenotypic methods . Six of the isolates were also identified by a species-specific polymerase chain reaction . Fourteen of the isolates were examined for resistance to a panel of seven antimicrobial agents using a disc diffusion method . All 18 isolates were serotyped according to the Page scheme using reference antisera in a haemagglutination inhibition test . RESULTS: Four of the 18 isolates were obtained from indigenous (kampung) chickens, with the remainder being from typical intensive poultry production systems . The 18 isolates were obtained from 11 outbreaks that showed the typical clinical signs of infectious coryza and 11 of the isolates were obtained from chickens that had been vaccinated with infectious coryza vaccines . All 18 isolates were confirmed as H paragallinarum by biochemical testing and six isolates were also identified as H paragallinarum by the polymerase chain reaction test . Eleven isolates were resistant to erythromycin and streptomycin, 10 to neomycin, eight to oxytetracycline, five isolates to doxycycline, three to sulphamethoxazoltrimethoprim but only one to ampicillin . Seven isolates were Page serovar A, four were Page serovar B and seven were Page serovar C . CONCLUSION: The presence of all three Page serovars (A, B and C) has been confirmed for the first time in Indonesian chickens . As the majority of the infectious coryza vaccines in use in Indonesia contain only serovar A and C, the presence of serovar B in chickens indicates that the protection by these bivalent vaccines would be reduced . The use of trivalent infectious coryza vaccines that contain serovars A, B and C is recommended for use in Indonesia.

J Korean Med Sci, 2000 Dec, 15(6), 616 - 22
Serotypes and antimicrobial susceptibility in clinical isolates of Haemophilus influenzae from Korean children in prevaccination era; Kwak YH et al.; Fifty-five strains of Haemophilus influenzae recovered at a children's hospital in Korea from 1992 through 1997, were analyzed for serotype and antibiotic resistance . Antimicrobial susceptibility was tested by broth dilution method . Among the 55 strains, 26 were from normally sterile body fluids, of which 17 were from the immunocompetent children . Spectrum in the immunocompetent included meningitis (47%), bacteremic pneumonia (18%), and bacteremia without focus (35%) . Three (12%) of 26 invasive infections were caused by non-type b: one type d and two type f . Nine of 29 non-sterile body fluid isolates belonged to one of encapsulted serotypes: four a, two c, one of each of b, d and e . Thirty two (58%) strains were resistant to ampicillin, and all of which produced beta-lactamase . All of the strains were highly susceptible to amoxicillin/clavulanate, cefixime, cefuroxime, azithromycin and ciprofloxacin, while 1 (2%), 7 (13%), 4 (7%) and 4 (7%) strains were intermediate to cefprozil, cefaclor, loracarbef, and clarithromycin, respectively . The serotype distribution of H . influenzae in Korean children is similar to those in developed countries before the introduction of Hib conjugate vaccine, and ampicillin resistance rate is among the highest published to date.

Kansenshogaku Zasshi, 2000 Dec, 74(12), 1081 - 7
{A case of pyogenic vertebral osteomyelitis caused by Haemophilus aphrophilus}; Nagoya H et al.; On Aug . 3, 1999, a 73-year-old male was admitted to our hospital with the chief complaint of pain in the neck, high fever, and numbness in the arm . MRI of the cervix showed high intensity at the C3/C4 disc space . Laboratory data showed several signs of inflammation . Haemophilus aphrophilus was detected from the specimen of the disc space, and the diagnosis of pyogenic vertebral osteomyelitis caused by H . aphrophilus was made . After the identification of H . aphrophilus, antibiotic therapy with Cefotiam (2 g/day) was given but his vertebral collapsed . Surgical treatment consisted of curettage and anterior spinal body fusion using the iliac bone, was performed on his 23rd hospital day, successfully . The antibiotic therapy of Cefazolin (2 g/day) was continued for the first 3 days, followed by Cefotiam (2 g/day) and later Levofloxacin (300 mg/day) . The patient was discharged on the 88th hospital day . The origin of infecting H . aphrophilus in this patient was not clear, but oral source was suspected . We reported the first case of pyogenic vertebral osteomyelitis caused by H . aphrophilus in Japan.

J Trop Pediatr, 2000 Dec, 46(6), 331 - 4
Haemophilus influenzae type b still remains a leading cause of meningitis among unvaccinated children--a prospective CSF analysis study; Uduman SA et al.; A prospective, hospital-based cerebrospinal fluid (CSF) analysis study was undertaken in 65 children who had diagnostic lumbar puncture on admission for suspected central nervous system infections . Twenty-three children were clinically diagnosed to have had sepsis and/or meningitis . CSF bacterial culture grew Haemophilus influenzae type b (Hib) in four cases and Streptococcus pneumonia (SP) was cultured in another child . Bacterial antigen was detected in 13 other CSF specimens and the pathogens were Hib (n = 9), SP (n = 3) and Group B Streptococcus (n = 1) . No etiologic cause was identified to explain the abnormal CSF pleocytosis and biochemistry in the remaining five cases . In contrast, the CSF analysis was normal in 42 other children with probable viral and non-infectious neurological condition, mostly febrile convulsions . The overall frequency rate for all types of meningitis and especially for Hib meningitis were 43 and 31 cases per 100,000 children < 5 years of age, respectively . These findings support our earlier observations that Hib meningitis still remains the leading cause of childhood meningitis in our region . Also it reaffirms the observation that bacterial meningitis may often be under-reported if CSF positive culture alone is considered for the diagnosis.

Thorax, 2001 Feb, 56(2), 109 - 14
Prospective study of the incidence, aetiology and outcome of adult lower respiratory tract illness in the community; Macfarlane J et al.; BACKGROUND: Acute lower respiratory tract illness in previously well adults is usually labelled as acute bronchitis and treated with antibiotics without establishing the aetiology . Viral infection is thought to be the cause in most cases . We have investigated the incidence, aetiology, and outcome of this condition . METHODS: Previously well adults from a stable suburban population consulting over one year with a lower respiratory tract illness were studied . For the first six months detailed investigations identified predetermined direct and indirect markers of infection . Evidence of infection was assessed in relation to presenting clinical features, indirect markers of infection, antibiotic use, and outcome . RESULTS: Consultations were very common, particularly in younger women (70/1000 per year in previously well women aged 16-39 years), mainly in the winter months; 638 patients consulted, of whom 316 were investigated . Pathogens were identified in 173 (55%) cases: bacteria in 82 (Streptococcus pneumoniae 54, Haemophilus influenzae 31, Moraxella catarrhalis 7), atypical organisms in 75 (Chlamydia pneumoniae 55, Mycoplasma pneumoniae 23), and viruses in 61 (influenza 23) . Seventy nine (24%) had indirect evidence of infection . Bacterial and atypical infection correlated with changes in the chest radiograph and high levels of C reactive protein but not with (a) the GP's clinical assessment of whether infection was present, (b) clinical features other than focal chest signs, and (c) outcome, whether or not appropriate antibiotics were prescribed . CONCLUSIONS: Over 50% of patients have direct and/or indirect evidence of infection, most commonly bacterial and atypical pathogens, but the outcome is unrelated to the identified pathogens . Many patients improve without antibiotics and investigations do not help in the management of these patients . GPs can reassure patients of the causes and usual outcome of this self-limiting condition.

Med Sci Monit, 2000 Mar-Apr, 6(2), 300 - 4
Antibiotic-sensitivity of Moraxella catarrhalis isolated from clinical materials in 1997-1998; Mikucka A et al.; Moraxella catarrhalis (M . catarrhalis) may normally be found in the upper respiratory tract . This bacterium, however, may cause infections such as acute otitis media, sinusitis, conjunctivitis, bronchitis chronica, pneumonia, endocarditis, septicaemia and meningitis . Haemophilus influenzae, Streptococcus pneumoniae and M . catarrhalis were the main causative agents responsible for respiratory tract infections . The major resistance problems associated with these species are those which cause resistance to beta-lactams . beta-lactamase was produced by > 80% M . catarrhalis strains . The susceptibility to ampicillin, amoxicillin/clavulanic acid, cefuroxime, erythromycin, ciprofloxacin was tested in 137 M . catarrhalis strains . All the strains resistant to ampicillin produced beta-lactamase and were sensitive to amoxicillin/clavulanic acid . For M . catarrhalis, the most active antimicrobials included cefuroxime (99%), ciprofloxacin (99%) and erythromycin (93%).

An Esp Pediatr, 2001 Feb, 54(2), 178 - 80
{Bacterial tracheitis due to Haemophilus influenzae}; Porta Ribera R et al.; We present a case of bacterial tracheitis in a 6.5 year old girl . Clinical signs and symptoms consisted of severe croup with high grade fever, which were preceded by upper respiratory tract prodrome . Initial treatment with steroids and nebulized epinephrine was unsuccessful . The patient was intubated a few hours after admission . Thick purulent secretions emerging from the trachea and the normal appearance of the epiglottis suggested the diagnosis of bacterial tracheitis, which was confirmed by isolation of Haemophilus influenzae in the culture of the tracheal secretions . The patient was administered a 14 day course of endovenous ceftriaxone and was kept on mechanical ventilation for 7 days . Fever and purulent tracheal secretions continued for the next 5 days . After 48 hours without these signs, laryngotracheobronchoscopy ruled out residual obstruction . Extubation was successfully performed . Fourteen days later physical examination showed no abnormalities and the patient was discharged . No complications were found during followup . The clinical, diagnostic and therapeutic aspects of this potentially life threatening entity that should taken into account in the differential diagnosis of severe croup are discussed.

Clin Infect Dis, 2001 Feb 15, 32(4), 566 - 72 Epub 2001 Feb 09.
Fever interval before diagnosis, prior antibiotic treatment, and clinical outcome for young children with bacterial meningitis; Bonsu BK et al.; In young children, meningitis due to Streptococcus pneumoniae is preceded by a long interval from onset of fever to diagnosis of bacterial meningitis (hereafter known as "fever interval"), during which time the patient frequently contacts a clinician . By means of retrospective chart review, we compared the fever interval that preceded diagnosis with the complication rate among 288 young children (age, 3--36 months) who had bacterial meningitis (1984--1996), as stratified by causative organism and prior antibiotic treatment . Pathogens included S . pneumoniae, Haemophilus influenzae type b, and Neisseria meningitidis . Pneumococcus species were associated with the longest fever interval prior to diagnosis of meningitis, the highest frequency of contact with a clinician before hospitalization, and the highest rate of documented morbidity or mortality . For S . pneumoniae, there was an association between antibiotic treatment received at prior meetings with a clinician and a reduced rate of meningitis-related complications (odds ratio, 0.14; P=.02) . Antibiotic treatment during such meetings is associated with a substantial reduction in disease-related sequelae.

Gene, 2001 Jan 10, 262(1-2), 169 - 77
Identification and characterization of an in vivo regulated D15/Oma87 homologue in Shigella flexneri using differential display polymerase chain reaction; Robb CW et al.; Shigella genes expressed during infection likely contribute to adaptation and virulence in the host . Using differential display PCR (DDPCR), a cDNA fragment from Shigella flexneri serotype 5 that showed enhanced expression in a murine model was identified, cloned and sequenced . Enhanced expression was verified by RNA dot blot . The full-length gene was cloned using PCR and sequenced . The complete gene sequence was BLAST searched against GenBank, and exhibited strong homology to genes encoding Haemophilus influenzae D15 and Pasteurella multocida Oma87 protective outer membrane antigens . The S . flexneri gene putatively encodes a approximately 90-kDa protein and was termed oma90 . The deduced amino acid sequence from oma90 was analyzed and compared to the D15/Oma87 antigens . Additionally, oma90 mapped to a cluster of orthologous groups, and probably contains an ancient conserved domain . The chromosomal organization of oma90 was similar to that for H . influenzae and P . multocida as well as for other known homologues . Northern blot revealed that the oma90 transcript encoded only oma90 . This report represents the first description of a S . flexneri gene identified based on enhanced expression in the host . Furthermore, we report the first evidence demonstrating in vivo regulation of a member of the d15/oma87 gene family.

Infect Immun, 2001 Mar, 69(3), 1938 - 42
Expression of cytolethal distending toxin and hemolysin is not required for pustule formation by Haemophilus ducreyi in human volunteers; Young RS et al.; Haemophilus ducreyi makes cytolethal distending toxin (CDT) and hemolysin . In a previous human challenge trial, an isogenic hemolysin-deficient mutant caused pustules with a rate similar to that of its parent . To test whether CDT was required for pustule formation, six human subjects were inoculated with a CDT mutant and parent at multiple sites . The pustule formation rates were similar at both parent and mutant sites . A CDT and hemolysin double mutant was constructed and tested in five additional subjects . The pustule formation rates were similar for the parent and double mutant . These results indicate that neither the expression of CDT, nor that of hemolysin, nor both are required for pustule formation by H . ducreyi in humans.

Infect Immun, 2001 Mar, 69(3), 1747 - 54
Expression of C-reactive protein in the human respiratory tract; Gould JM et al.; C-reactive protein (CRP) is a normal constituent of human sera synthesized by hepatocytes and induced by proinflammatory cytokines . The function of this acute-phase reactant includes activation of complement and enhancement of opsonophagocytosis . CRP binds to phosphorylcholine (ChoP), a constituent of eukaryotic membranes that is also found on the cell surface of major bacterial pathogens of the human respiratory tract, including Streptococcus pneumoniae and Haemophilus influenzae . The presence of CRP on mucosal surfaces and role in innate immunity in the human respiratory tract where ChoP-containing organisms reside have not been previously studied . We have shown using a monoclonal antibody to CRP that CRP is present in inflamed (0.17 to 42 microg/ml) and uninflamed (<0.05 to 0.88 microg/ml) secretions from the human respiratory tract in sufficient quantities for an antimicrobial effect . In addition, the CRP gene was expressed in epithelial cells of the human respiratory tract using in situ hybridization on nasal polyps and reverse transcriptase PCR of pharyngeal cells in culture . The complement-dependent bactericidal activity of normal nasal airway surface fluid and sputum against ChoP-expressing H . influenzae was abolished when the secretions were pretreated to remove CRP . In summary, the results indicate that CRP is present in secretions of the human respiratory tract, that human respiratory epithelial cells are capable of CRP expression, and that this protein may contribute to bacterial clearance in the human respiratory tract.

Infect Immun, 2001 Mar, 69(3), 1650 - 60
Haemophilus somnus induces apoptosis in bovine endothelial cells in vitro; Sylte MJ et al.; Haemophilus somnus causes pneumonia, reproductive failure, infectious myocarditis, thrombotic meningoencephalitis, and other diseases in cattle . Although vasculitis is commonly seen as a result of systemic H . somnus infections, the pathogenesis of vascular damage is poorly characterized . In this study, we demonstrated that H . somnus (pathogenic isolates 649, 2336, and 8025 and asymptomatic carrier isolates 127P and 129Pt) induce apoptosis of bovine endothelial cells in a time- and dose-dependent manner, as determined by Hoechst 33342 staining, terminal deoxynucleotidyl transferase-mediated dUTP-FITC nick end labeling, DNA fragmentation, and transmission electron microscopy . H . somnus induced endothelial cell apoptosis in as little as 1 h of incubation and did not require extracellular growth of the bacteria . Viable H . somnus organisms induced greater endothelial cell apoptosis than heat-killed organisms . Since viable H . somnus cells release membrane fibrils and blebs, which contain lipooligosaccharide (LOS) and immunoglobulin binding proteins, we examined culture filtrates for their ability to induce endothelial cell apoptosis . Culture filtrates induced similar levels of endothelial cell apoptosis, as did viable H . somnus organisms . Heat inactivation of H . somnus culture filtrates partially reduced the apoptotic effect on endothelial cells, which suggested the presence of both heat-labile and heat-stable factors . We found that H . somnus LOS, which is heat stable, induced endothelial cell apoptosis in a time- and dose-dependent manner and was inhibited by the addition of polymyxin B . These data demonstrate that H . somnus and its LOS induce endothelial cell apoptosis, which may play a role in producing vasculitis in vivo.

Infect Immun, 2001 Mar, 69(3), 1488 - 91
DsrA-deficient mutant of Haemophilus ducreyi is impaired in its ability to infect human volunteers; Bong CT et al.; Haemophilus ducreyi produces an outer membrane protein called DsrA, which is required for serum resistance . An isogenic dsrA mutant, FX517, was constructed previously in H . ducreyi 35000 . Compared to its parent, FX517 cannot survive in normal human serum . When complemented in trans with a plasmid containing dsrA, FX517 is converted to a serum-resistant phenotype (C . Elkins, K . J . Morrow, Jr., and B . Olsen, Infect . Immun . 68:1608-1619, 2000) . To test whether dsrA was transcribed in vivo, we successfully amplified transcripts in five biopsies obtained from four experimentally infected human subjects . To test whether DsrA was required for virulence, six volunteers were experimentally infected with 35000 and FX517 and observed for papule and pustule formation . Each subject was inoculated with two doses (70 to 80 CFU) of live 35000 and 1 dose of heat-killed bacteria on one arm and with three doses (ranging from 35 to 800 CFU) of live FX517 on the other arm . Papules developed at similar rates at sites inoculated with the mutant or parent . However, mutant papule surface areas were significantly smaller than parent papules . The pustule formation rate was 58% (95% confidence interval {CI} of 28 to 85%) at 12 parent sites, and 0% (95% CI of 0 to 15%) at 18 mutant sites (P = 0.0004) . Although biosafety regulations precluded our testing the complemented mutant in humans, these results suggest that expression of DsrA facilitates the ability of H . ducreyi to progress to the pustular stage of disease.

Infect Immun, 2001 Mar, 69(3), 1483 - 7
Transcription of candidate virulence genes of Haemophilus ducreyi during infection of human volunteers; Throm RE et al.; Haemophilus ducreyi expresses several putative virulence factors in vitro . Isogenic mutant-to-parent comparisons have been performed in a human model of experimental infection to examine whether specific gene products are involved in pathogenesis . Several mutants (momp, ftpA, losB, lst, cdtC, and hhdB) were as virulent as the parent in the human model, suggesting that their gene products did not play a major role in pustule formation . However, we could not exclude the possibility that the gene of interest was not expressed during the initial stages of infection . Biopsies of pustules obtained from volunteers infected with H . ducreyi were subjected to reverse transcription-PCR . Transcripts corresponding to momp, ftpA, losB, lst, cdtB, and hhdA were expressed in vivo . In addition, transcripts for other putative virulence determinants such as ompA2, tdhA, lspA1, and lspA2 were detected in the biopsies . These results indicate that although several candidate virulence determinants are expressed during experimental infection, they do not have a major role in the initial stages of pathogenesis.

Pediatr Dev Pathol, 2001 Mar-Apr, 4(2), 105 - 21
Asplenic-hyposplenic overwhelming sepsis: postsplenectomy sepsis revisited; Hansen K et al.; Absence of the spleen or splenic function predisposes individuals to risk of overwhelming infection . These infections are most often due to encapsulated organisms, especially pneumococcus, Haemophilus influenzae type b, and meningococcus, but any bacterial agent may cause the rapid onset of septicemia, meningitis, pneumonia, and shock characteristic of the asplenic-hyposplenic condition . The risk is greatest in infants and young children, but asplenic-hyposplenic adults also have an increased risk of infection . Prophylactic antibiotics and immunization with polyvalent pneumococcal, H . influenzae type b, and meningococcal vaccines have reduced the incidence of infections in asplenic-hyposplenic individuals, but even these measures have not eliminated the risk . Surgeons have adopted techniques to save as much splenic tissue as possible and some splenic functions, such as pitting red cells, have been preserved, but conservative surgery has not provided total protection against overwhelming infection . Therapies designed to interrupt the cascade of overwhelming sepsis have not yet been successful . In those cases in which the spleen is surgically removed, the underlying disease or condition leading to splenectomy influences the risk of sepsis . Splenectomy incidental to other operations, such as gastrectomy, results in the lowest risk for overwhelming infection, but this is still some 35-fold greater than the risk for overwhelming infections in the general population . In increasing order of risk, the other main indications for surgical removal of the spleen are idiopathic thrombocytopenia purpura, trauma, transplantation procedures, hereditary spherocytosis, staging Hodgkin's disease, portal hypertension with hypersplenism, and thalassemia . Pathologists should comment on the risk of overwhelming sepsis when spleens are processed as surgical specimens, and should carefully weigh all splenic tissue, including accessory spleens and splenic implants (splenosis), in autopsy cases with and without overwhelming sepsis.

Expert Opin Investig Drugs, 2001 Feb, 10(2), 353 - 67
Telithromycin: a new ketolide antimicrobial for treatment of respiratory tract infections; Yassin HM et al.; Telithromycin is a new ketolide antimicrobial, specifically developed for the treatment of community-acquired respiratory tract infections . It has a wide spectrum of antibacterial activity against common respiratory pathogens including Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pyogenes . It also has activity against atypical pathogens, such as Chlamydia pneumoniae, Legionella pneumophila and Mycoplasma pneumoniae . Telithromycin maintains activity against beta-lactam and macrolide-resistant respiratory tract pathogens and does not appear to induce cross-resistance to other members of the macrolide-lincosamide-streptogramin (MLS) group of antimicrobials . It demonstrates bactericidal activity against S . pneumoniae and H . influenzae and has a prolonged concentration-dependent post-antibiotic effect (PAE) in vitro . The drug has favourable pharmacokinetics following oral administration . It is well absorbed, achieves good plasma levels and is highly concentrated in pulmonary tissues and white blood cells . In clinical trials, telithromycin given orally at a dose of 800 mg once daily for 5 - 10 days was as effective as comparator antimicrobials for the treatment of adults with community-acquired pneumonia, acute exacerbations of chronic bronchitis, acute maxillary sinusitis and group A-beta-haemolytic streptococcal pharyngitis or tonsillitis . The adverse events and safety profile were similar to comparator antimicrobials . The most common adverse events were diarrhoea, nausea, headache and dizziness . Telithromycin should provide an effective, convenient and well-tolerated once-daily oral therapy for treatment of respiratory infections.

Arch Otolaryngol Head Neck Surg, 2001 Feb, 127(2), 180 - 3
Preventing labyrinthitis ossificans: the role of steroids; Hartnick CJ et al.; OBJECTIVE: To identify a possible relationship between the administration of steroids at the time of diagnosis of bacterial meningitis and the development of labyrinthitis ossificans . DESIGN: Retrospective analysis of the charts of 38 children requiring cochlear implantation who presented with bacterial meningitis and then developed bilateral profound deafness . The patients' charts were reviewed for age at diagnosis, the type of antibiotic administered, and the administration, dosage, and duration of steroid (dexamethasone) therapy . Labyrinthitis ossificans was established by preoperative computed tomographic and/or magnetic resonance imaging and by the intraoperative findings as described in the operative report . PATIENTS AND METHODS: Patients were 38 children who received cochlear implantation by a single senior otolaryngologist for bacterial meningitis-related deafness . Ten patients' charts (26%) were available for full review; 9 of these 10 patients had documented pneumococcal meningitis and the other patient had Haemophilus influenzae-type meningitis . RESULTS: One of the 6 patients who received steroid therapy at the time of initial illness had documented evidence of labyrinthitis ossificans either radiographically or at the time of surgery . All 4 patients who failed to receive steroid therapy developed labyrinthitis ossificans . The results achieve statistical significance by chi2 analysis and a t test (P<.01) . CONCLUSION: The results of this retrospective study are highly suggestive of a role for steroids in preventing the development of labyrinthitis ossificans in children with pneumococcal meningitis.

Pediatr Infect Dis J, 2001 Jan, 20(1), 116 - 9; discussion 120-2
Efficacy of ofloxacin and other otic preparations for acute otitis media in patients with tympanostomy tubes; Goldblatt EL; Otorrhea occurs in 21 to 50% of all children with tympanostomy tubes in the United States . More than 1 million children annually undergo tubomyringotomy, constituting placement of more than 2 million tympanostomy tubes each year . The organisms typically responsible for otorrhea are the same as those that cause otitis media in very young children, including Streptococcus pneumonia, Haemophilus influenzae and Moraxella catarrhalis . Drainage from tympanostomy tubes in older children involves organisms that colonize the external auditory canal, the most common being Pseudomonas aeruginosa and Staphylococcus aureus . Ofloxacin (Floxin otic), a newer fluoroquinalone antibiotic, has several advantages over other agents available for the treatment of otorrhea caused by acute otitis media in patients with tympanostomy tubes . The twice daily dosing regimen encourages better patient adherence to therapy, which is likely to improve treatment efficacy . Ofloxacin has not been associated with ototoxicity in animal models or in children participating in the clinical trials . It provides coverages for a wide range of pathogens, including Pseudomonas sp., and is indicated for use in children > or =1 year old and currently approved for patients > or =12 years with chronic suppurative otitis media . Ofloxacin applied topically in children with tympanostomy tubes in place and purulent otorrhea is as efficacious as oral amoxicillin/clavulanate (Augmentin) therapy . Other currently available therapeutic options are discussed.

Pediatr Infect Dis J, 2001 Jan, 20(1), 52 - 8
Seroetiology of acute lower respiratory infections among hospitalized children in Beijing; Yang Y et al.; BACKGROUND: Little is known of the etiology of childhood acute lower respiratory infections in China, where the use of antimicrobials is indiscriminate . Trials to change such a policy require etiologic data, especially on the bacteria most relevant to these common diseases . METHODS: One hundred consecutive infants and children from 3 months to 14 years of age with symptoms and signs compatible with acute lower respiratory infections were studied prospectively in the largest pediatric hospital in Beijing from February to May, 1997 . Blood culture, thorax radiography and paired sera for 20 microbiologic assays were taken, and the course of illness was monitored uniformly . Disease severity was graded . RESULTS: In 24 cases there was evidence only of bacterial etiology, and in 5 solely viral agents were found; 3 children probably had a mixed bacterial-viral infection . Surprisingly no pneumococcal infection was detected, Mycoplasma pneumoniae (n = 21), Haemophilus influenzae type b (n = 8) and Chlamydia pneumoniae (n = 7) being the dominant bacteria . All children recovered . CONCLUSIONS: Routine use of antimicrobials for these patients seems unjustified . Serologic evidence for the H . influenzae type b etiology is encouraging in terms of vaccination, but confirmatory studies are needed.

Curr Opin Pediatr, 2001 Feb, 13(1), 70 - 4
Pneumococcal conjugate vaccine; Ledwith M; Pneumococcal infections account for a significant proportion of bacterial infections in infants and children . The growing threat from pneumococci resistant to penicillin and other antimicrobials has led to increased pressure for the development of an effective vaccine . The only vaccine available until recently, a purified polysaccharide vaccine, is limited in that it fails to induce adequate and long-lasting immunity in infants under 2 years of age, the age group most at risk from this disease . Polysaccharide antigens conjugated to certain proteins induce effective immunity with a rapid response to subsequent infection or antigen challenge . The success of the protein-conjugated haemophilus influenzae vaccine supports the strategy of protein-conjugated polysaccharide vaccines . Currently, published trials of conjugated pneumococcal vaccines have shown the effectiveness and safety of these vaccines . Conjugate vaccines also provide protection against otitis media and may eliminate nasopharyngeal carriage of this organism . Widespread use of this vaccine is both cost effective and safe.

Diagn Microbiol Infect Dis, 2001 Jan, 39(1), 49 - 53
In vitro activity of linezolid (U-100766) against Haemophilus influenzae measured by three different susceptibility testing methods; Biedenbach DJ et al.; Linezolid has excellent antibacterial activity against a wide range of Gram-positive organisms . Early in vitro investigations suggested that the compound also had activity against some Gram-negative species, including those commonly associated with community-acquired respiratory tract infections (Haemophilus influenzae and Moraxella catarrhalis) . Against 603 recent clinical isolates of H . influenzae from geographically diverse regions of the world, tested by the reference broth microdilution method (HTM), linezolid MIC values ranged from 2-64 microg/ml (MIC50 and MIC90 at 16 microg/ml and 32 microg/ml, respectively) . A subset of 328 strains was also tested to compare broth microdilution and Etest (AB BIODISK, Solna, Sweden) methodologies . The Etest method produced slightly higher MIC results attributable to a growth-enhancing effect of the 5% CO2 incubation used in the test procedure and pH changes . Linezolid activity versus H . influenzae was limited as evidenced by reference test results (susceptible breakpoints at < or = 2 or < or = 4 microg/ml) and variable endpoints were obtained when alternative methods were used such as the Etest or standardized disk diffusion procedures . Clinical laboratories should limit the testing of linezolid against Gram-negative species (H . influenzae).

N Engl J Med, 2001 Feb 8, 344(6), 403 - 9
Efficacy of a pneumococcal conjugate vaccine against acute otitis media; Eskola J et al.; BACKGROUND: Ear infections are a common cause of illness during the first two years of life . New conjugate vaccines may be able to prevent a substantial portion of cases of acute otitis media caused by Streptococcus pneumoniae . METHODS: We enrolled 1662 infants in a randomized, double-blind efficacy trial of a heptavalent pneumococcal polysaccharide conjugate vaccine in which the carrier protein is the nontoxic diphtheria-toxin analogue CRM197 . The children received either the study vaccine or a hepatitis B vaccine as a control at 2, 4, 6, and 12 months of age . The clinical diagnosis of acute otitis media was based on predefined criteria, and the bacteriologic diagnosis was based on a culture of middle-ear fluid obtained by myringotomy . RESULTS: Of the children who were enrolled, 95.1 percent completed the trial . With the pneumococcal vaccine, there were more local reactions than with the hepatitis B vaccine but fewer than with the combined whole-cell diphtheria-tetanus-pertussis and Haemophilus influenzae type b vaccine that was administered simultaneously . There were 2596 episodes of acute otitis media during the follow-up period between 6.5 and 24 months of age . The vaccine reduced the number of episodes of acute otitis media from any cause by 6 percent (95 percent confidence interval, -4 to 16 percent {the negative number indicates a possible increase in the number of episodes}), culture-confirmed pneumococcal episodes by 34 percent (95 percent confidence interval, 21 to 45 percent), and the number of episodes due to the serotypes contained in the vaccine by 57 percent (95 percent confidence interval, 44 to 67 percent) . The number of episodes attributed to serotypes that are cross-reactive with those in the vaccine was reduced by 51 percent, whereas the number of episodes due to all other serotypes increased by 33 percent . CONCLUSIONS: The heptavalent pneumococcal polysaccharide-CRM197 conjugate vaccine is safe and efficacious in the prevention of acute otitis media caused by the serotypes included in the vaccine.

Clin Microbiol Infect, 2000 Apr, 6(4), 185 - 94
Meropenem: clinical response in relation to in vitro susceptibility; Drusano GL et al.; OBJECTIVE: To collate the clinical response and pathogen eradication rates for meropenem monotherapy with in vitro susceptibility of the causative pathogens . METHODS: Data were compiled from 17 randomized clinical studies that compared meropenem monotherapy with standard treatment options, often combinations . A total of 4906 pathogens from lower respiratory tract, intra-abdominal, obstetric/gynecological, skin/soft tissue, meningitis, or pediatric infections were assessed . Of these, 3713 pathogens (1963 meropenem, 1750 comparators) were evaluable . RESULTS: The overall rates of satisfactory clinical response (cure or improvement) and pathogen eradication (eradication or presumed eradication) at the end of therapy were similar with meropenem (93% for both responses) and the comparators (92%), as were the rates in each infection type . For each pathogen, the clinical response and eradication rates with meropenem were similar across the minimum inhibitory concentration (MIC) range of < or = 0.25 to 4 mg/L . Overall, a satisfactory clinical response occurred in 93% (1580 of 1708) of infections caused by nonfastidious pathogens with MICs < or = 4 mg/L and in 84% (16 of 19) of those with an MIC of 8 mg/L . Pathogen eradication rates were similar (93 and 79%, respectively) . A similar profile was observed for fastidious pathogens . The high rates of satisfactory clinical response and pathogen eradication produced by meropenem in each type of infection were generally independent of the causative pathogen, whether Gram-positive or -negative aerobe or anaerobe or when occurring as mono- or polymicrobial infections . CONCLUSIONS: The attractive in vitro profile of meropenem translates into good clinical efficacy . The National Committee for Clinical Laboratory Standards has now defined meropenem MIC breakpoints for nonfastidious aerobes or anaerobes as < or = 4 (susceptible), 8 (intermediate) and > or = 16 mg/L (resistant), respectively . The susceptibility breakpoint for Streptococcus spp . (excluding Streptococcus pneumoniae) is < or = 0.5 mg/L and, since meropenem is indicated for the treatment of meningitis, the susceptibility breakpoint for S . pneumoniae and Haemophilus influenzae is < or = 0.25 and < or = 0.5 mg/L, respectively . Meropenem monotherapy is therefore a valid option for the initial empirical treatment of a range of serious infections caused by single or multiple bacterial pathogens.

Clin Microbiol Infect, 2000 Apr, 6(4), 178 - 84
Antimicrobial susceptibility of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis collected from five centers in Brazil, 1997-98; Critchley IA et al.; OBJECTIVE: To assess the susceptibility of the key respiratory pathogens Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis to antimicrobial agents used to treat respiratory tract infections . METHODS: Isolates were collected from five centers in Brazil during 1997-98, and susceptibility testing was conducted at a central laboratory according to National Committee for Clinical Laboratory Standards criteria . RESULTS: Of the 359Streptococcus pneumoniae isolates tested, 77% were susceptible, 19% were intermediate and 4% were resistant to penicillin . The susceptibility of S . pneumoniae to other beta-lactams and macrolides was greater than 90%, but cotrimoxazole was active against only 48% of the isolates . The prevalence of susceptible isolates was 100.0% for vancomycin and 99.7% for levofloxacin . beta-Lactam, macrolide, and cotrimoxazole activities were negatively associated with penicillin resistance . Of the 219 isolates of Haemophilus influenzae tested, 11% produced beta-lactamase and 11% were not susceptible to ampicillin . Nearly all H . influenzae isolates were susceptible to all other drugs, except cotrimoxazole (47% susceptibility) . Of the 52 Moraxella catarrhalis isolates, 98% produced beta-lactamase, and the MIC of all drugs was </=4 mg/L, with the exception of ampicillin, where the MIC90 was> 8 mg/L . CONCLUSIONS: When these data are compared with previous reports, our findings suggest that the prevalence of pneumococci that are resistant to agents such as penicillin and cotrimoxazole may be increasing in Brazil, which highlights the need to continue surveillance programs.

Clin Microbiol Infect, 2000 Oct, 6(10), 519 - 24
Antibiotic susceptibility and genotypic characterization of Haemophilus influenzae strains isolated from nasopharyngeal specimens from children in day-care centers in eastern France; Talon D et al.; OBJECTIVE: To determine the overall carriage rate for Haemophilus influenzae in young children in day-care centers, the frequency of resistance to various classes of antibiotic, and the clonal relationship between isolates of the various resistant phenotypes . METHODS: Nasopharyngeal (NP) specimens were obtained and cultured on chocolate agar with bacitracin . Antibiotic susceptibility testing and serotyping were performed for all isolates . The genetic polymorphism of ampicillin-susceptible and beta-lactamase-producing isolates was studied by pulsed-field gel electrophoresis using SmaI . RESULTS: Of the 596 NP secretion cultures, 152 (25.5%) were positive for H . influenzae . Sixty-four (42.1%) isolates produced beta-lactamase and two (1.3%) were ampicillin resistant but did not produce beta-lactamase . We were unable to serotype 150 isolates; one isolate belonged to capsular serotype e and one to serotype f . Forty-six major DNA patterns were identified among 76 randomized isolates . beta-lactamase producing isolates more frequently showed EP than ampicillin-susceptible isolates P < 10(-4) . The frequency of isolates with EP was significantly lower in day-care centers attended by less than 20 children than in those attended by more than 20 children (P = 0.020) . CONCLUSIONS: Resistance due to beta-lactamase production has disseminated in some day-care centers, mostly by person-to-person spread but also via the possible conjugal transfer of large plasmids between strains . The size of day-care centers may affect the risk of transmission.

Acta Ophthalmol Scand, 2000 Dec, 78(6), 694 - 8
Bacteriology and antibiotic therapy in congenital nasolacrimal duct obstruction; Kuchar A et al.; AIMS: To determine the current bacteriology of mucopurulent discharge in congenital nasolacrimal duct obstruction (CNDO), the in vitro response to different antibiotics and clinical effectiveness of the antibiotics used to relieve babies from mucopurulent discharge . METHODS: A clinical study evaluated the effectiveness of local antibiotic agents clinically and in vitro . 50 samples were obtained from the lacrimal sac in 47 young children with CNDO . The patients' mean age was 21.45 +/- 17.09 months . The cultures were incubated and the infectious agents isolated . Sensitivity testing was performed in each case, testing 10 different local antibiotics . A control group of 10 babies expected for cataract surgery was constituted . RESULTS: Cultures were positive for bacteria from 72.64% of the samples . 73 isolates were recovered from the 50 samples . The bacterial species most frequently cultured was Streptococcus pneumoniae, representing 35.4% of the isolates, followed by Haemophilus influencae (19.6%) . The sensitivity testing revealed ofloxacin and tetracycline to be the most effective drugs as monotherapy . Clinically the combination of bacitracin and neomycin, primarily used in half of the patients as initial therapy, was successful in curing the dacryocystitis in 82.5% of all patients . CONCLUSION: Chronic dacryocystitis due to CNDO is associated with an equal proportion of Gram positive and negative bacteria, which can be treated with a high effectiveness by a combination drug of bacitracin and neomycin.

Vaccine, 2001 Feb 8, 19(13-14), 1671 - 7
Antibody responses to pneumococcal and haemophilus vaccinations in patients with B-cell chronic lymphocytic leukaemia; Hartkamp A et al.; Although vaccination against Streptococcus pneumoniae (S . pneumoniae) and Haemophilus influenzae type b (Hib) is recommended for immunocompromised patients, such as patients with B-cell chronic lymphocytic leukaemia (B-CLL), its protective effect is questionable . We studied antibody responses to pneumococcal polysaccharide vaccine (Pneumovax-23) and to conjugated H . influenzae type b-vaccine (Act-Hib) in 25 patients with B-CLL . After vaccination, the number of patients with antibody levels in the protective range against pneumococcal serotypes and H . influenzae b increased from 9 (38%) to 12 (50%) of 24 patients and from 8 (35%) to 11 (48%) of 23 patients, respectively . The patients with adequate antibody response to Pneumovax-23 and Act-Hib had significantly less advanced stages of B-CLL, higher gammaglobulin levels, total IgG-levels and IgG-subclasses 2 and 4 levels, and lower levels of soluble CD23 . Consequently, vaccination with these vaccines should be given as soon as the diagnosis of B-CLL is made, early in the course of the disease with determination of post-vaccination antibody levels.

Int J Antimicrob Agents, 2001 Feb, 17(2), 93 - 6
Changes in strategies for optimal antibacterial therapy in cystic fibrosis; Ratjen F; Aggressive antibiotic therapy of bacterial airway infection is one of the main reasons for the dramatic increase in life expectancy over the last few decades . Staphylococcus aureus and Haemophilus influenzae are the predominant pathogens in younger patients, but the choice of antibiotic therapy against these pathogens remains highly controversial . There is general agreement that patients with pulmonary exacerbations should be treated and many cystic fibrosis (CF) centres will also try to eradicate bacteria in the absence of symptoms . Prophylactic antibiotic therapy, with anti-staphylococcal medications started at the time of diagnosis, is advocated by some groups but its positive effect remains unproven . In fact, recent studies have suggested that continuous prophylactic treatment with anti-staphylococcal antibiotics may increase the risk of early colonisation with Pseudomonas aeruginosa . P . aeruginosa is the main pathogen in older children with CF . While chronic airway infection with mucoid P . aeruginosa is considered irreversible, both the combination of oral ciprofloxacin with inhaled colistin and inhaled tobramycin alone has been used successfully in the early phase of colonisation . In patients chronically infected with P . aeruginosa, standard treatment of pulmonary exacerbations consists of intravenous combination therapy for 2-3 weeks . Controversy exists whether this treatment should be performed routinely every 3 months or only in the presence of a pulmonary exacerbation . Inhaled antibiotics such as tobramycin have been shown to improve lung function and reduce sputum density of P . aeruginosa, but both the optimal dose and the duration of therapy are unclear at the present time.

Gene, 2000 Dec 23, 259(1-2), 207 - 15
The orientation bias of Chi sequences is a general tendency of G-rich oligomers; Uno R et al.; The Chi sequences are specific oligomers that stimulate DNA repair by homologous recombination, and are different sequences in each organism . Approximately 75% of the copies of the Chi sequence (5'-GCTGGTGG-3') of Escherichia coli reside on the leading strand, and this orientation bias is often believed to be a consequence of the biological role of Chi sequences as the signal sequence of RecBCD pathway in DNA replication . However, our computer analysis found that many G-rich oligomers also show this asymmetric orientation pattern . The shift in the Chi orientation bias appears around the replication origin and terminus, but these locations are also coincident with the shift points in GC content or GC skew . We conducted the same analysis with the genome of Bacillus subtilis, and found that in addition to Chi, other G-rich oligomers show similar asymmetric orientation patterns, whose shift points were coincident with those of the GC skew . However, the genome of Haemophilus influenzae Rd, whose GC skew is not so pronounced, does not clearly show asymmetric orientation patterns of Chi or other G-rich oligomers . These results lead us to suggest that the uneven distribution of the Chi orientation between the two strands of the double helix is mostly due to the uneven distribution of G content (GC skew) and that the replication-related function of Chi sequences is not the primary factor responsible for the evolutionary pressure causing the orientation bias.

Vaccine, 2000 Dec 8, 19 Suppl 1, S148 - 52
Human lactoferrin proteolytic activity: analysis of the cleaved region in the IgA protease of Haemophilus influenzae; Plaut AG et al.; Human lactoferrin proteolytically cleaves and inactivates two colonization factors of non-typable Haemophilus influenzae, the IgA protease precursor protein (Iga), and Hap, the non-pilus adhesin by which microoganisms adhere to host epithelial cells and form microcolonies . Iga and Hap are homologous proteins that are members of the autotransporter family of secreted proteins expressed by gram-negative bacteria . Studies of Iga cleaved by lactoferrin, reported here, show that proteolysis occurred within the helper region of Iga (Iga(beta)) domain which anchors the autotransporter within the Haemophilus outer membrane . The amino-terminus of the extracted Iga protein was not modified . The location of the proteolytic active site in human lactoferrin is under study . Lactoferrin proteolysis may attenuate pathogenicity of H . influenzae, an important cause of otitis media.

Vaccine, 2000 Dec 8, 19 Suppl 1, S144 - 7
Xylitol in preventing acute otitis media; Uhari M et al.; Xylitol is a polyol sugar alcohol and is referred to as birch sugar, because it can be produced from birch . Natural sources of xylitol include plums, strawberries, raspberries and rowan berries . Xylitol inhibits the growth of Streptococcus pneumoniae and it inhibits the attachment of both pneumococci and Haemophilus influenzae on the nasopharyngeal cells . In two clinical trials xylitol was found efficient to prevent the development of acute otitis media with a daily dose of 8.4-10 g of xylitol given in five divided doses . The efficacy in these 2-3 months follow-up trials was approximately 40% when chewing gum was used and approximately 30% with xylitol syrup . The need to use antimicrobials reduced markedly when using xylitol . In a high-risk group of children with tympanostomy tubes xylitol was ineffective in preventing otitis . Xylitol appears to be an attractive alternative to prevent acute otitis media . A more practical frequency of doses should be found before its use can be widely recommended.

Vaccine, 2000 Dec 8, 19 Suppl 1, S116 - 21
Passive immunization for the prevention of otitis media; Englund JA et al.; The safety and protective efficacy of exogenously-administered immunoglobulin for the prevention of otitis media has been demonstrated in the clinical trials of the human-derived polyclonal immune globulin used to prevent Haemophilus influenzae type b disease and respiratory syncytial virus infection in high risk neonates and young children . However, this form of therapy is expensive, difficult to administer due to the requirements of slow intravenous infusion or relatively large volumes given intramuscularly, and associated with side effects related to the volume and nature of the immunoglobulin preparation . In contrast, RSV-specific monoclonal antibody has not been as successful as human-derived immunoglobulin in preventing otitis media in high risk infants . The administration of monoclonal-antibody for the prevention of otitis media will be difficult, potentially due to the need for antibody to multiple epitopes of the viral and bacterial pathogens which could be targets . The use of maternal antibody to provide passive immunity to young infants at a time when they are most vulnerable to severe sequelae of infection can also be considered . We have studied maternal immunization using either a 23-valent pneumococcal polysaccharide vaccine or a conjugate H . influenzae type b (Hib) vaccine . Significant levels of maternally-derived Hib or pneumococcal antibody were transferred from the mother to the infant at the time of birth and persisting, for some antigens, through 2 months of age . The use of maternal immunization to prevent otitis media and other respiratory complications remains to be studied, but results of these small clinical trials indicate further clinical investigation is warranted.

Vaccine, 2000 Dec 8, 19 Suppl 1, S108 - 15
Developing a nontypeable Haemophilus influenzae (NTHi) vaccine; Poolman JT et al.; There is a current high demand for nontypable Haemophilus influenzae (NTHi) vaccines . Various options for the composition of such vaccines are possible . Decisions about the vaccine composition have to take into account the antigenic variability of NTHi, so even complex immunogens such as whole bacteria would preferentially have a tailor-made antigenic composition . We will present a summary of NTHi vaccine development, describing research efforts from SmithKline Beecham and other laboratories . Currently, major (P1, P2, P4, P5) and minor (P6, D15, TbpA/B, ellipsis) outer membrane proteins, LPS, adhesins (HMW, Hia, pili, P5) are being studied . Preclinical results with LPD, P5 (LB1) and OMP26 from our laboratories will be described including the use of animal models of otitis and lung infection.

Vaccine, 2000 Dec 8, 19 Suppl 1, S41 - 50
The pathogenesis of nontypable Haemophilus influenzae otitis media; St Geme JW 3rd; Nontypable Haemophilus influenzae is a common cause of otitis media and initiates infection by colonizing the upper respiratory tract . In this article, I review our current understanding of the molecular determinants of H . influenzae colonization and discuss the relationship between colonization and otitis media.

Vaccine, 2000 Dec 8, 19 Suppl 1, S9 - S16
Treatment of acute otitis media - challenges in the era of antibiotic resistance; Dagan R; The last decade is characterized by the increase in antibiotic resistance among respiratory bacterial pathogens in the presence of only modest progress in the development of new antibacterial agents to overcome this resistance . A series of recent studies show clearly that the increased resistance among the main AOM pathogens (namely Streptococcus pneumoniae and Haemophilus influenzae) is associated with a dramatic decrease in bacteriologic response to antibiotic treatment, which in turn has an impact on clinical response . Thus, the individual patient is affected by the increasing antibiotic resistance . Moreover, the society as a whole is now also affected because the carriage and spread of antibiotic resistant AOM pathogens is remarkably impacted by antibiotic treatment . New studies show the remarkable ability of antibiotics to rapidly promote nasopharyngeal carriage and spread of antibiotic-resistant AOM pathogens . In these studies, the increase in carriage of antibiotic resistant S . pneumoniae is shown already after 3-4 days from initiation of antibiotic treatment and may last for weeks to months after treatment . Children carrying antibiotic-resistant organisms transmit those organisms to their family and to their day care centers and thus a vicious cycle is created in which increased antibiotic resistance with decreased response leads to increased antibiotic use, which in turn leads to further increase in resistance . New antibiotics are not likely to improve this situation . It is clear that the challenge in the next decade is to prevent AOM rather than to treat it . Efforts to prevent AOM include improved environmental factors, immunization with bacterial and viral vaccines and some creative measures such as prevention of colonization and attachment to epithelium of AOM pathogens . Whether these efforts will prove successful or, even if successful, will only modify the clinical and bacteriologic picture presenting new challenges, only time will tell.

Biochem Biophys Res Commun, 2001 Jan 12, 280(1), 81 - 4
The influence of ATP on the association and unfolding of the tyrosine repressor ligand response domain of Haemophilus influenzae; Kristl S et al.; The secondary structure of the ligand response domain of the Haemophilus influenzae tyrosine repressor, TyrR(lrd), was investigated using CD spectroscopy which revealed 42.5% alpha-helix, 17.6% beta-sheet, and 39.9% loops . Quaternary structure analysis by fluorescence anisotropy showed that TyrR(lrd) is monomeric at a concentration of 100 nM to 2 microM but that the protein readily dimerizes in the presence of its natural ligand ATP . Equilibrium unfolding studies of TyrR(lrd) using guanidinium hydrochloride suggested a two-state model with no detectable stable intermediates . The unfolding transition monitored by CD spectroscopy was responsive to tyrosine and ATP resulting in a shift to higher denaturant concentrations in the presence of these ligands . Differential scanning calorimetry yielded melting temperatures, T(m), of 51.15 and 58.07 degrees C for the unliganded and for the ATP-liganded protein, respectively . ATP is thus proposed to be a major structural cofactor for the molecular architecture of TyrR(lrd) .

J Nutr, 2001 Feb, 131(2), 336S - 9S
Genetic research and nutritional individuality; Eckhardt RB; Recent genetic research builds on a base established over the last century by physicians and nutritional scientists, who introduced the concept of biochemical individuality and documented its significance for understanding a wide variety of problems in human health . Current comparative genomic investigations on a variety of organisms (Haemophilus influenzae, Saccharomyces cerevisiae, Caenorhabditis elegans, Drosophila melanogaster, Homo sapiens) have established the existence of numerous orthologs (proteins in different organisms that show significant sequence similarities over 80% of their lengths), suggesting significant conservation of structure and probably some of function as well . At the same time, molecular comparisons among individuals within our own species show the existence of abundant molecular variants, many of which have been shown to have functional significance in nutritional and related metabolic contexts . The combination of biochemical individuality and known functional utilities of allelic variants should converge to create a situation in which nutritional optima can be specified as part of comprehensive lifestyle prescriptions tailored to the needs of each person.

J Bacteriol, 2001 Mar, 183(5), 1585 - 94
Nucleotide sequence and analysis of conjugative plasmid pVT745; Galli DM et al.; The complete nucleotide sequence and genetic map of pVT745 are presented . The 25-kb plasmid was isolated from Actinobacillus actinomycetemcomitans, a periodontal pathogen . Two-thirds of the plasmid encode functions related to conjugation, replication, and replicon stability . Among potential gene products with a high degree of similarity to known proteins are those associated with plasmid conjugation . It was shown that pVT745 derivatives not only mobilized a coresident nontransmissible plasmid, pMMB67, but also mediated their own conjugative transfer to different A . actinomycetemcomitans strains . However, transfer of pVT745 derivatives from A . actinomycetemcomitans to Escherichia coli JM109 by conjugation was successful only when an E . coli origin of replication was present on the pVT745 construct . Surprisingly, 16 open reading frames encode products of unknown function . The plasmid contains a conserved replication region which belongs to the HAP (Haemophilus-Actinobacillus-Pasteurella) theta replicon family . However, its host range appears to be rather narrow compared to other members of this family . Sequences homologous to pVT745 have previously been detected in the chromosomes of numerous A . actinomycetemcomitans strains . The nature and origin of these homologs are discussed based on information derived from the nucleotide sequence.

J Bacteriol, 2001 Mar, 183(5), 1540 - 51
Expression of the Moraxella catarrhalis UspA1 protein undergoes phase variation and is regulated at the transcriptional level; Lafontaine ER et al.; The UspA1 protein of Moraxella catarrhalis has been shown to function as an adhesin that mediates adherence to human epithelial cell lines in vitro (E . R . Lafontaine, L . D . Cope, C . Aebi, J . L . Latimer, G . H . McCracken, Jr., and E . J . Hansen, J . Bacteriol . 182:1364-1373, 2000) . In the present study, cell lysates prepared from individual colonies of several M . catarrhalis wild-type strains were analyzed by Western blot analysis using monoclonal antibodies (MAbs) specific for the UspA1 protein . Expression of UspA1 was shown to exhibit phase variation that was correlated with both adherence ability in vitro and the number of guanine (G) residues contained within a homopolymeric {poly(G)}tract located upstream of the uspA1 open reading frame (ORF) . Nucleotide sequence analysis revealed that isolates expressing relatively high levels of UspA1 had 10 G residues in their uspA1 poly(G)tracts, whereas isolates that expressed much lower levels of UspA1 had 9 G residues . This poly(G) tract was located 30 nucleotides (nt) upstream of the uspA1 ORF and 168 nt downstream of the uspA1 transcriptional start site . Primer extension experiments, RNA slot blot analysis, and cat reporter constructs were used to demonstrate that M . catarrhalis isolates with 10 G residues in their uspA1 poly(G) tracts expressed two-to threefold more uspA1 mRNA than did isolates which had 9 G residues in their poly(G)tracts . Northern hybridization analysis revealed that an intact uspA1 mRNA was readily detectable in RNA from M . catarrhalis isolates that had 10 G residues in their uspA1 poly(G) tracts, whereas no full-length uspA1 mRNA was observed in isolates whose poly(G)tracts contained 9 G residues . M . catarrhalis strain O35E uspA1 genes that contained wild-type and mutated poly(G) tracts were expressed in Haemophilus influenzae to demonstrate that the length and composition of the poly(G)tract affected expression of UspA1.

Infect Immun, 2001 Feb, 69(2), 773 - 8
Antibodies to loop 6 of the P2 porin protein of nontypeable Haemophilus influenzae are bactericidal against multiple strains; Neary JM et al.; The P2 porin protein is the most abundant protein in the outer membrane of nontypeable Haemophilus influenzae (NTHI) . Analysis of sequences of P2 from different strains reveals the presence of both heterogeneous and conserved surface-exposed loops of the P2 molecule among strains . The present study was undertaken to test the hypothesis that antibodies to a conserved surface-exposed loop are bactericidal for multiple strains of NTHI and could thus form the basis of vaccines to prevent infection due to NTHI . Polyclonal antiserum to a peptide corresponding to loop 6 was raised and was immunopurified over a loop 6 peptide column . Analysis of the antibodies to whole organisms and peptides corresponding to each of the eight loops of P2 by immunoassays revealed that the antibodies were highly specific for loop 6 of P2 . The immunopurified antibodies bound to P2 of 14 of 15 strains in immunoblot assays . These antibodies to loop 6 demonstrated complement-mediated bactericidal killing of 8 of 15 strains . These results support the concept of using conserved regions of the P2 protein as a vaccine antigen.

Infect Immun, 2001 Feb, 69(2), 695 - 705
Serum resistance in an invasive, nontypeable Haemophilus influenzae strain; Williams BJ et al.; A common feature of many different organisms causing bacteremia is the ability to avoid the bactericidal effects of normal human serum . In Haemophilus influenzae encapsulated strains are particularly serum resistant; however, we found that a nonencapsulated strain (R2866) isolated from the blood of an immunocompetent child with meningitis who had been successfully immunized with H . influenzae type b conjugate vaccine was serum resistant . Since serum resistance usually involves circumventing the action of the complement system, we defined the deposition of various complement components on the surfaces of this H . influenzae strain (R2866), a nonencapsulated avirulent laboratory strain (Rd), and a virulent type b encapsulated strain (Eagan) . Membrane attack complex (MAC) accumulation correlated with the loss of bacterial viability; correspondingly, the rates of MAC deposition on the serum-sensitive strain Rd and the serum-resistant strains differed . Analysis of cell-associated immunoglobulin G (IgG), C1q, C3b, and C5b indicated that serum-resistant H . influenzae prevents MAC accumulation by delaying the synthesis of C3b through the classical pathway . Among the initiators of the classical pathway, IgG deposition contributes most of the C3 convertase activity necessary to start the cascade ending with MAC deposition . Despite similar IgG binding, strain R2866 delays C3 convertase activity compared to strain Rd . We conclude that strain R2866 can persist in the bloodstream, in part by inhibiting or delaying C3 deposition on the cell surface, escaping complement mediated killing.

Bioinformatics, 2000 Nov, 16(11), 968 - 77
Short interrupted palindromes on the extragenic DNA of Escherichia coli K-12, Haemophilus influenzae and Neisseria meningitidis; Vasconcelos AT et al.; MOTIVATION: The importance of the various kinds of repetitive nucleotide sequences for the workings of bacterial DNA has been widely recognized . This work is concerned with the distribution of a particular group of repetitive sequences, the short-sequenced interrupted extragenic palindromes, on the genetic maps of Escherichia coli K-12, Haemophilus influenzae Rd and Neisseria meningitidis Z2491 and MC58 . A tool has been developed based upon a statistical hypothesis test taking into account the markovian structure of random sequences in order to determine the non-random character of extragenic palindromes . RESULTS: Totals of 7631, 12904, 4722 and 5477 non-random short interrupted palindromes have been found on the E.coli, H.influenzae, and N.meningitidis serogroup A and serogroup B genomes, respectively . Their distribution patterns on the respective genomes vary according to the bacterial species considered . Based on their position on the genome, palindromes could be distinguished as those which integrate longer, repetitive sequences; those which stand in isolation, and still others are associated to specific genome sites . AVAILABILITY: The complete list of the observed palindromes is available at the site CONTACT: atrv@lncc.br

J Clin Microbiol, 2001 Feb, 39(2), 601 - 5
Diagnosing genital ulcer disease in a clinic for sexually transmitted diseases in Amsterdam, The Netherlands; Bruisten SM et al.; The most common etiologic agents of genital ulcer disease (GUD) are herpes simplex virus type 1 (HSV-1), HSV-2, Treponema pallidum, and Haemophilus ducreyi . In an outpatient clinic for sexually transmitted diseases in Amsterdam, The Netherlands, specimens from 372 patients with GUD were collected from February to November 1996 . Sera were collected at the time of the symptoms and, for most patients, also during follow-up visits . Swabs in viral transport medium were used for HSV culture and for detection of DNA . The most prevalent pathogen found was HSV-2, which was detected by culture in 35% of the patients and by PCR in 48% of the patients . Also, HSV-1 infection was more often detected by PCR (7.8%) than by culture (5.6%) . Evidence for an active infection with T . pallidum was found in 1.9% of the patients, using serological tests . A multiplex PCR for simultaneous T . pallidum and H . ducreyi DNA detection was positive for T . pallidum in 3.3% of the samples and for H . ducreyi in only 0.9% (3 out of 368) of the samples . The sensitivity of the PCR was superior to that of culture for HSV detection and to that of serology for T . pallidum detection . Specific H . ducreyi immunoglobulin G antibodies were detected in sera of 5.2% of the patients, with no concordance between serology and PCR . In 37% of the cases, none of the tested microorganisms was detected . Performance of PCR in addition to conventional techniques significantly improved the diagnosis of GUD.

J Bacteriol, 2001 Feb, 183(4), 1168 - 74
Identification of a plasmid-encoded gene from Haemophilus ducreyi which confers NAD independence; Martin PR et al.; Members of the family Pasteurellaceae are classified in part by whether or not they require an NAD supplement for growth on laboratory media . In this study, we demonstrate that this phenotype can be determined by a single gene, nadV, whose presence allows NAD-independent growth of Haemophilus influenzae and Actinobacillus pleuropneumoniae . This gene was cloned from a 5.2-kb plasmid which was previously shown to be responsible for NAD independence in Haemophilus ducreyi . When transformed into A . pleuropneumoniae, this cloned gene allowed NAD-independent growth on complex media and allowed the utilization of nicotinamide in place of NAD on defined media . Sequence analysis revealed an open reading frame of 1,482 bp that is predicted to encode a protein with a molecular mass of 55,619 Da . Compared with the sequence databases, NadV was found to have significant sequence homology to the human pre-B-cell colony-enhancing factor PBEF and to predicted proteins of unknown function identified in the bacterial species Mycoplasma genitalium, Mycoplasma pneumoniae, Shewanella putrefaciens, Synechocystis sp., Deinococcus radiodurans, Pasteurella multocida, and Actinobacillus actinomycetemcomitans . P . multocida and A . actinomycetemcomitans are among the NAD-independent members of the Pasteurellaceae . Homologues of NadV were not found in the sequenced genome of H . influenzae, an NAD-dependent member of the Pasteurellaceae, or in species known to utilize a different pathway for synthesis of NAD, such as Escherichia coli . Sequence alignment of these nine homologues revealed regions and residues of complete conservation that may be directly involved in the enzymatic activity . Identification of a function for this gene in the Pasteurellaceae should help to elucidate the role of its homologues in other species.

J Antimicrob Chemother, 2001 Feb, 47(2), 211 - 4
Individual use of antibiotics and prevalence of beta-lactamase production among bacterial pathogens from middle ear fluid; Thrane N et al.; Prescription data and clinical laboratory data were analysed to assess the influence of previous antibiotic therapy on the prevalence of beta-lactamase in isolates of Haemophilus influenzae and Moraxella catarrhalis from primary specimens of middle ear fluid from 2129 children aged 0-5 years . The prevalence of beta-lactamase-positive H . influenzae was 6.6% {95% confidence interval (CI): 3.5-9.8%} in children who received antibiotics 5-90 days before isolation of the organism compared with 7.0% (95% CI: 3.9-10.2%) in those who did not . The prevalence of beta-lactamase-positive M . catarrhalis was 90.9% (95% CI: 84.0-97.8%) in children who received antibiotics compared with 86.7% (95% CI: 79.0-94.4%) in those who did not.

Clin Microbiol Rev, 2000 Apr, 13(2), 302 - 17
Worldwide Haemophilus influenzae type b disease at the beginning of the 21st century: global analysis of the disease burden 25 years after the use of the polysaccharide vaccine and a decade after the advent of conjugates; Peltola H; Vaccination against Haemophilus influenzae type b (Hib) diseases began a quarter of a century ago with a polysaccharide vaccine; this vaccine was followed by four different conjugates 10 years later . In this review, the burden of global Hib disease is quantified following this 25-year period of vaccine availability to determine the potential impact of conjugate vaccines . This task was accomplished by analysis of data available in 10 languages in 75 geographical regions of over 50 countries . All severe Hib diseases, not only meningitis, were characterized, and special attention was paid to the most vulnerable age group, i.e., children aged 0 to 4 years . Prior to vaccination, the weighted worldwide incidence of meningitis in patients younger than 5 years was 57/100,000, and for all Hib diseases except nonbacteremic pneumonia, it was 71/100,000, indicating 357,000 and 445,000 cases per year, respectively . At least 108,500 of these children died . For all age groups combined, there were 486,000 cases of Hib disease, excluding pneumonia, with 114,200 deaths and probably an equal number of sequelae per annum . If the figures for nonbacteremic pneumonia are included, a conservative estimate is that over 2.2 million cases of infection and 520,000 deaths from Hib disease occurred worldwide, but the true numbers might have been greater . Despite these large numbers and availability of safe and efficacious vaccines, only 38,000 cases annually are prevented-a meager 8% or less than a 2% reduction in cases, depending on whether nonbacteremic pneumonia is included in the calculations . Although vaccination has had great success in some affluent countries, the current level of activity has had a very small impact globally . The use of conjugates, preferably with a reduced number of doses and in combination with other vaccines or perhaps in fractional doses, should be extended to less privileged countries, where most Hib disease occurs.

Br J Cancer, 2000 Apr, 82(7), 1261 - 5
Immunogenicity of vaccination against influenza, Streptococcus pneumoniae and Haemophilus influenzae type B in patients with multiple myeloma; Robertson JD et al.; Vaccination against influenza and Streptococcus pneumoniae is recommended for elderly and immunocompromised individuals . However, there is little information concerning the efficacy of vaccination in specific groups of patients . In this study, 52 patients underwent vaccination against influenza, S . pneumoniae and Haemophilus influenzae type b (Hib) as they attended hospital outpatient clinics . Serum was analysed prior to vaccination and 4-6 weeks afterwards . Antibody titres against S . pneumoniae and Hib were compared with reference values corresponding to the geometric mean titres of a healthy UK population . For influenza vaccination, haemagglutination inhibition (HI) titres were measured against three inactivated strains; a titre of > or = 1/40 was considered protective . No patient had protective titres to all three antigens prior to vaccination and 41 patients (85%) had titres < 1/40 to all 3 strains . Post vaccination only 9/48 patients (19%) achieved protective antibody titres . Resistance to S . pneumoniae and response to Pneumovax II was also poor: prevaccination, 45 patients (93%) had suboptimal antibody titres and in 26/43 patients (61%) titres remained low post vaccination . Resistance to Hib and response to vaccination was comparable with the healthy adult UK population . These results question the practice of routine influenza and pneumococcal vaccination in myeloma patients.

J Immunol, 2000 Apr 15, 164(8), 4185 - 96
Haemophilus influenzae stimulates ICAM-1 expression on respiratory epithelial cells; Frick AG et al.; Epithelial cells interact directly with bacteria in the environment and play a critical role in airway defense against microbial pathogens . In this study, we examined the response of respiratory epithelial cells to infection with nontypable Haemophilus influenzae . Using an in vitro cell culture model, we found that epithelial cell monolayers released significant quantities of IL-8 and expressed increased levels of ICAM-1 mRNA and surface protein in response to H . influenzae . In contrast, levels of IL-1beta, TNF-alpha, and MHC class I were not significantly affected, suggesting preferential activation of a specific subset of epithelial genes directed toward defense against bacteria . Induction of ICAM-1 required direct bacterial interaction with the epithelial cell surface and was not reproduced by purified H . influenzae lipooligosaccharide . Consistent with a functional role for this response, induction of ICAM-1 by H . influenzae mediated increased neutrophil adherence to the epithelial cell surface . Furthermore, in an in vivo murine model of airway infection with H . influenzae, increased epithelial cell ICAM-1 expression coincided with increased chemokine levels and neutrophil recruitment in the airway . These results indicate that ICAM-1 expression on human respiratory epithelial cells is induced by epithelial cell interaction with H . influenzae and suggest that an ICAM-1-dependent mechanism can mediate neutrophil adherence to these cells independent of inflammatory mediator release by other cell types . Direct induction of specific epithelial cell genes (such as ICAM-1 and IL-8) by bacterial infection may allow for rapid and efficient innate defense in the airway.

J Mol Evol, 2000 Mar, 50(3), 264 - 75
Evolutionary lability of context-dependent codon bias in bacteria; McVean GA et al.; In bacteria, synonymous codon usage can be considerably affected by base composition at neighboring sites . Such context-dependent biases may be caused by either selection against specific nucleotide motifs or context-dependent mutation biases . Here we consider the evolutionary conservation of context-dependent codon bias across 11 completely sequenced bacterial genomes . In particular, we focus on two contextual biases previously identified in Escherichia coli; the avoidance of out-of-frame stop codons and AGG motifs . By identifying homologues of E . coli genes, we also investigate the effect of gene expression level in Haemophilus influenzae and Mycoplasma genitalium . We find that while context-dependent codon biases are widespread in bacteria, few are conserved across all species considered . Avoidance of out-of-frame stop codons does not apply to all stop codons or amino acids in E . coli, does not hold for different species, does not increase with gene expression level, and is not relaxed in Mycoplasma spp., in which the canonical stop codon, TGA, is recognized as tryptophan . Avoidance of AGG motifs shows some evolutionary conservation and increases with gene expression level in E . coli, suggestive of the action of selection, but the cause of the bias differs between species . These results demonstrate that strong context-dependent forces, both selective and mutational, operate on synonymous codon usage but that these differ considerably between genomes.

Genome Res, 2001 Jan, 11(1), 28 - 42
Gene expression changes triggered by exposure of Haemophilus influenzae to novobiocin or ciprofloxacin: combined transcription and translation analysis; Gmuender H et al.; The responses of Haemophilus influenzae to DNA gyrase inhibitors were analyzed at the transcriptional and the translational level . High-density microarrays based on the genomic sequence were used to monitor the expression levels of >80% of the genes in this bacterium . In parallel the proteins were analyzed by two-dimensional electrophoresis . DNA gyrase inhibitors of two different functional classes were used . Novobiocin, as a representative of one class, inhibits the ATPase activity of the enzyme, thereby indirectly changing the degree of DNA supercoiling . Ciprofloxacin, a representative of the second class, obstructs supercoiling by inhibiting the DNA cleavage-resealing reaction . Our results clearly show that different responses can be observed . Treatment with the ATPase inhibitor Novobiocin changed the expression rates of many genes, reflecting the fact that the initiation of transcription for many genes is sensitive to DNA supercoiling . Ciprofloxacin mainly stimulated the expression of DNA repair systems as a response to the DNA damage caused by the stable ternary complexes . In addition, changed expression levels were also observed for some genes coding for proteins either annotated as "unknown function" or "hypothetical" or for proteins not directly involved in DNA topology or repair.

J Chemother, 2000 Dec, 12(6), 503 - 8
Comparative activity of cefodizime and ceftriaxone against respiratory pathogens in an in vitro pharmacodynamic model simulating concentration-time curves; Blandino G et al.; The duration of time that serum levels are above the minimum inhibitory concentration (MIC; T >MIC) seems to be an important pharmacodynamic parameter for beta-lactams . The aim of this study was to evaluate the bactericidal activity of cefodizime and ceftriaxone in a pharmacokinetic model mimicking the concentrations in bronchial mucus and in serum (total and free) obtained at 2, 4, 8, 12 and 24 h, after 1 g i.m . administration once daily . The species investigated were respiratory pathogens (1 strain of Staphylococcus aureus, 2 strains of Streptococcus pneumoniae, 1 strain b-lactamase negative and 1 strain beta-lactamase positive of Haemophilus influenzae, 1 strain of Escherichia coli and 1 strain of Klebsiella pneumoniae); MIC50s of the chosen strains were reported . In this in vitro model the concentrations (serum and bronchial mucus) for both antibiotics are generally at or above the MIC values of the tested strains until 24 hours . The killing curve showed rapid killing for both antibiotics: 99.9% killing (a 3-log reduction in growth) within 6 to 8 h, depending upon the microorganism tested . There was no significant difference in the log kill between cefodizime and ceftriaxone . These data confirm that T >MIC for beta-lactams is the pharmacodynamic parameter which best correlates with bactericidal efficacy . On the basis of the killing curve determined for cefodizime versus ceftriaxone at concentrations that these antibiotics can reach during therapy with 1 g i.m . once daily we expect reasonable clinical efficacy with monoadministration of cefodizime as well as for ceftriaxone in respiratory tract infections.

J Biomed Mater Res, 2001, 58(1), 113 - 20
Bioactive glass S53P4 in repair of septal perforations and its interactions with the respiratory infection-associated microorganisms Haemophilus influenzae and Streptococcus pneumoniae; Stoor P et al.; Interpositional grafts between mucoperiosteal flaps are commonly used in the repair of septal perforations . We studied the use of bioactive glass (BAG) S53P4 as an interpositional graft in 11 patients suffering from septal perforations . In aqueous environments, ions are released from the BAG and the pH rises in its vicinity, both of which may influence the growth and adhesion of microorganisms . Thus, we also studied the effects of the BAG S53P4 as granules or discs on the respiratory infection-associated microorganisms Haemophilus influenzae and Streptococcus pneumoniae . Growth inhibition was studied using an agar plate test and adhesion was analyzed both with and without serum precoating of the BAG S53P4 . The perforations were successfully closed in 10 of 11 patients . One patient had a near total septum perforation, which could not be closed . No BAG-associated infections were seen during the follow-up . The BAG S53P4 did not show any clear growth inhibition of the microorganisms, which showed low adhesion to the material . Serum precoating increased the adsorption . Thus, uncoated BAG S53P4 seems to be a good graft in the repair of septal perforations .

Hybridoma, 2000 Dec, 19(6), 445 - 53
Production and characterization of a new monoclonal antibody against Neisseria meningitidis: study of the cross-reactivity with different bacterial genera; De Gaspari EN; We have generated a hybridoma cell line which produces an 8C7Br1 clone of the IgM antibody isotype . It recognizes the 50-, 65-, and 60-kDa antigens and is reactive with strains of N . meningitidis in the 98% of local Neisseria genera by Dot-ELISA assays . Two percent of the strains of N . meningitidis B do not present reactivity with the 8C7Br1 monoclonal antibody (MAb) . The antibody reacted against N . meningitidis of serogroups A, B, C, X, Y, Z, and different serotypes and subtypes of N . meningitidis B and C by means of Dot-ELISA and Immunoblot . It cross-reacted with Neisseria gonorrhoeae, Neisseria lactamica, Haemophilus influenzae type b, Escherichia coli, Salmonella typhimurium, Salmonella typhi, Shigella flexneri, Bordetella pertussis, and Bacillus subtilis . The 8C7Br1 MAb reacted with the 65-kDa protein present in the prototype meningococcal strains B:16:B6(B2a:P1.5.2) and 2996 (B2b:P1.5.2) . In H . influenzae type b, E . coli and B . subtilis, the MAb recognized the protein of 60, 65, and 70 kDa, respectively . FACS analysis showed that 8C7Brl MAb could recognize the 50-kDa protein on the surface of N . meningitidis homologous (B:4:P1.9) strain . These results, together with the bactericidal activity of 8C7Br1, and an experiment of passive protection in mice, demonstrated the potential importance of the cross-reactive protein as a candidate antigen for N . meningitidis B vaccine composition.

World Health Organ Tech Rep Ser, 2000, 897, i - vi, 1-106
WHO Expert Committee on Biological Standardization . Forty-ninth report.
{Microorganisms isolated in cases of pertussis-like syndrome}
Ferrer A, Calico I, Manresa JM, Andreu A, Moraga F, Valle I.

Servei de Microbiologia, Hospital Vall d'Hebron, Pg Vall d'Hebron 119-129, 08035 BarcelonaOBJECTIVE: To describe the etiologic study of the pertussis-like syndrome, not only as far as Bordetella genus is concerned but also regarding the causative role of other microorganisms for a 11-year period (1988-1998) . METHODS: In all specimens from patients suffering from pertussis-like cough the presence of Bordetella spp., other bacteria, viruses, and mycoplasma was investigated . The analysed data included microbiological findings and epidemiologic issues (age, sex, hospital admission area, yearly distribution and seasonal period) . RESULTS: A total of 1,063 specimens were investigated, most of them nasopharyngeal aspirates (910), corresponding to 905 patients; a positive culture was obtained form 56.9 of these patients . B . pertussis was isolated from 10.5% of patients . As for other bacteria, Haemophilus influenzae and Streptococcus pneumoniae were also isolated, in 16.9% and 15.8% of occasions, respectively . The respiratory syncitial virus was isolated from 10.7% of patients and other viruses in 9.4% . Among mycoplasma, Ureaplasma urealyticum predominated, with a recovery rate of 2.9% . The male/female ratio was 495/410; the ages of 67.2% of patients ranged from 0 to 6 months; a total of 689 (76.1%) required hospital admission . The recovery of B . pertussis and adenoviruses predominated during spring and summer months . In contrast, H . influenzae, S . pneumoniae and respiratory syncitial virus were recovered more frequently during winter months . CONCLUSIONS: Most patients with pertussis-like syndrome are children aged less than 6 months . The recovery percentages of B . pertussis and respiratory syncitial virus are identical and therefore we think that the investigation of their presence in this syndrome is fully warranted as well as the search for other microorganisms, since clinical symptoms are commonly non-specific among infants.

Ann Pharmacother, 2000 Dec, 34(12), 1469 - 77
Cefdinir: an expanded-spectrum oral cephalosporin; Guay DR; OBJECTIVE: To review the antimicrobial activity, pharmacokinetics, clinical efficacy, and tolerability of cefdinir, an expanded-spectrum oral cephalosporin . DATA SOURCES: Literature was identified by a MEDLINE search (January 1983-November 1999) of the medical literature, review of English-language literature and bibliographies of these articles, and product information . STUDY SELECTION: Clinical efficacy data were selected from all published trials mentioning cefdinir . Additional information concerning in vitro susceptibility, safety, chemistry, and pharmacokinetic profile of cefdinir was also reviewed . DATA SYNTHESIS: Cefdinir, an oral expanded-spectrum cephalosporin, has a broad spectrum of activity against many gram-negative and -positive aerobic organisms, including Streptococcus pneumoniae, Staphylococcus aureus, Streptococcus pyogenes, Haemophilus influenzae, and Moraxella catarrhalis . Cefdinir is stable to hydrolysis by many common beta-lactamases . Cefdinir is rapidly absorbed from the gastrointestinal tract and is primarily eliminated via renal clearance of unchanged drug . The terminal disposition half-life of cefdinir is approximately 1.5 hours . Efficacy has been demonstrated in a number of clinical trials in adults and children with upper and lower respiratory tract infections (e.g., pharyngitis, sinusitis, acute otitis media, acute bronchitis, acute exacerbation of chronic bronchitis, community-acquired pneumonia) and skin and skin-structure infections . The adverse event profile is similar to that of comparator agents . CONCLUSIONS: Cefdinir is a second-line alternative to first-line antimicrobial agents, with convenient once- or twice-daily dosing in the treatment of upper and lower respiratory tract infections and skin and skin-structure infections . Similar to other oral expanded-spectrum cephalosporins, cefdinir has activity against common pathogens of the respiratory tract and skin and is stable in the presence of many beta-lactamases . The clinical choice of an oral expanded-spectrum cephalosporin will be based on patient acceptance, frequency of administration, and cost.

Pediatr Infect Dis J, 2000 Dec, 19(12 Suppl), S153 - 8
Five-day twice daily cefdinir therapy for acute otitis media: microbiologic and clinical efficacy; Block SL et al.; OBJECTIVE: To examine the microbiologic and clinical efficacy of a 5-day course of cefdinir in the treatment of tympanocentesis-documented acute otitis media (AOM) . DESIGN: Open label noncomparative trial . SETTING: Primary care, ambulatory . PATIENTS: Children ages 6 months through 12 years with signs of AOM and middle ear effusion confirmed by tympanometry in at least one ear . INTERVENTION: Patients underwent tympanocentesis at baseline and received cefdinir 7 mg/kg twice a day for 5 days . MAIN OUTCOME MEASURES: Presumptive eradication of middle ear pathogens determined by clinical cure of signs and symptoms of AOM at end of therapy (Study Days 7 to 9) and Visit 3 (Study Days 16 to 21) . RESULTS: A total of 125 of 177 enrolled children had 134 pathogens isolated by tympanocentesis: Streptococcus pneumoniae, 69 (51.5%); Haemophilus influenzae 44 (32.8%; beta-lactamase-positive in 18 of 44 strains); beta-lactamase-positive Moraxella catarrhalis, 15 (11.2%); and Streptococcus pyogenes, 6 (4.5%) . The clinical cure rates by patient in the microbiologically and overall clinically evaluable groups, respectively, were 73% (84 of 115) and 77.4% (130 of 168) at the end of therapy visit and 57.4% (66 of 115) and 61.9% (104 of 168) at Visit 3 . Presumptive eradication rates at end of therapy were 8 of 11 (72.7%) and 4 of 8 (50%) for patients with penicillin-intermediate and -resistant S . pneumoniae isolates, respectively . Adverse reactions occurred in 16% of patients, with diarrhea (11%) occurring most frequently . CONCLUSIONS: A 5-day regimen of cefdinir was effective in the eradication of the common causative pathogens of nonrefractory AOM, including intermediate penicillin-resistant S . pneumoniae and beta-lactamase-producing organisms . Cefdinir should be considered a suitable second line antibiotic for AOM.

Pediatr Infect Dis J, 2000 Dec, 19(12), 1135 - 40
Immunogenicity of a Haemophilus influenzae type b-tetanus toxoid conjugate vaccine when mixed with a diphtheria-tetanus-acellular pertussis-hepatitis B combination vaccine; Greenberg DP et al.; BACKGROUND: Combination vaccines are urgently needed to reduce the number of injections given to young children . The aim of the study was to evaluate the safety and immunogenicity of a combination vaccine that contains diphtheria and tetanus toxoids and acellular pertussis antigens (DTaP), recombinant hepatitis B surface antigen (HepB) and Haemophilus influenzae type b (Hib) polysaccharide conjugated to tetanus toxoid (PRP-T) . METHODS: Four hundred five infants were randomized equally to three groups and immunized at 2, 4 and 6 months of age with: (1) DTaP/HepB vaccine used to reconstitute lyophilized PRP-T vaccine and administered as a single injection; (2) DTaP/HepB vaccine and PRP-T vaccine administered as two separate injections; or (3) DTaP, HepB and PRP-T vaccines administered as three separate injections . Safety was closely monitored, and blood specimens were obtained to assess antibody responses to each vaccine antigen . RESULTS: All study vaccines were well-tolerated, and the rates of systemic and injection site reactions were similar between groups . After the third dose the geometric mean antibody concentrations to Hib were significantly lower in subjects in Group 1 (1.63 microg/ml) compared with subjects in Groups 2 and 3 (6.26 and 6.15 microg/ml, respectively; P < 0.0001) . Subjects with antibody concentrations <1.0 microg/ml after the third dose responded well to a booster dose of Hib conjugate vaccine given at 11 to 15 months of age (41 of 44 with anti-PRP > or = 1.0 microg/ml) . Differences between groups for antibody responses to the other vaccine components were not clinically significant . CONCLUSIONS: Infants given a combined DTaP/ HepB/PRP-T vaccine experienced a significantly lower antibody response to the PRP-T component than infants given PRP-T vaccine as a separate injection . However, the immune response to a booster dose of Hib conjugate vaccine indicated the presence of immunologic memory.

Pediatr Infect Dis J, 2000 Dec, 19(12), 1119 - 27
Immunogenicity and safety of a new liquid hexavalent combined vaccine compared with separate administration of reference licensed vaccines in infants; Mallet E et al.; OBJECTIVE: The immunogenicity and safety of a new liquid hexavalent vaccine (diphtheria-tetanus-acellular pertussis-inactivated polio vaccine-hepatitis B-polyribosyl ribitol phosphate conjugated to tetanus protein; Hexavac; Aventis Pasteur MSD, Lyon, France) are compared with those of reference vaccines {diphtheria-tetanus-acellular pertussis-inactivated polio vaccine reconstituting lyophilized purified Haemophilus influenzae polysaccharide conjugated to tetanus protein vaccine (Pentavac; Aventis Pasteur MSD) and hepatitis B vaccine (H-B-Vax II; Aventis Pasteur MSD)} injected separately at the same visit in a prospective multicenter, comparative, open label trial . METHODS: Infants were randomized to receive Hexavac (n = 423) or Pentavac and H-B-Vax II (n = 425) as a primary immunization series at 2, 4 and 6 months of age . Seroprotection and seroconversion rates against all antigens at 1 month after the primary series were compared between the two vaccine groups with 95% confidence intervals (CI0.95) and were considered clinically equivalent (not inferior) when the upper limit of the 95% confidence interval on the difference (reference, hexavalent) was below predefined differences . RESULTS: Hexavac met and surpassed the pre-defined criteria for clinical equivalence to Pentavac and H-B-Vax II given concomitantly . It elicited similar seroprotection and seroconversion rates against all antigens . Seroprotection and seroconversion rates obtained 1 month after the third dose of Hexavac were >90% for all antigens . The postimmunization antibody geometric mean titers (GMT) for hepatitis B and purified Haemophilus influenzae polysaccharide were about 2-fold higher in infants who received the reference vaccines than in infants who had received Hexavac . GMTs for poliovirus antibodies tended to be enhanced in infants vaccinated with Hexavac . GMTs for all other antigens were very similar among both groups . Hexavac was generally well-tolerated . At least one local reaction was reported in 20.3% of Hexavac injections compared with 15.8% at the Pentavac injections site and 3.8% at the H-B-Vax II injections site . These reactions were generally mild and transient . At least one systemic adverse event was reported in 45.7% of Hexavac injections compared with 42.2% of Pentavac and H-B-Vax II injections (mild fever, irritability and drowsiness were most frequently reported) . The frequency of adverse events was not significantly different between groups . No vaccine-related serious adverse event occurred during the study . CONCLUSION: This liquid hexavalent vaccine was generally well-tolerated and provided immune responses adequate to be protective against six infectious diseases with a single injection, given at 2, 4 and 6 months of age.

Schweiz Med Wochenschr, 2000, Suppl 125, 35S - 37S
{Epiglottitis--a pediatric disease?}; Schupbach J et al.; Epiglottitis, commonly described as a paediatric disease, also occurs in adults . Early diagnosis and immediate treatment are crucial because of the rapid and possibly lethal course of upper airway obstruction due to swelling . Initial treatment consists in securing the upper airway and in antibiotic treatment . Streptococci and, especially in children, Haemophilus influenzae b are the most common bacteria . Our study focused on clinical and epidemiological changes since children started to be vaccinated against Haemophilus influenzae b in Switzerland (1992) . We reviewed patient histories of 31 adults and 88 children who were hospitalised with epiglottitis at the University Hospital of Berne between 1989 and 1999 . Our findings show that the incidence of epiglottitis in children, a clinically, epidemiologically and bacteriologically homogeneous disease, has dramatically decreased . Epiglottitis in adults presents as a more heterogeneous disease without change since the beginning of the vaccination programme . Due to the variety of germs it is impossible to recommend vaccination for adults against Haemophilus influenzae b.

J Membr Biol, 2000 Dec 1, 178(3), 185 - 93
Charged residues in surface-located loops influence voltage gating of porin from Haemophilus influenzae type b; Arbing MA et al.; Porin of Haemophilus influenzae type b (341 amino acids; M(r) 37782) determines the permeability of the outer membrane to low molecular mass compounds . Purified Hib porin was subjected to chemical modification of lysine residues by succinic anhydride . Electrospray ionization mass spectrometry identified up to 12 modifications per porin molecule . Tryptic digestion of modified Hib porin followed by reverse phase chromatography and matrix assisted laser desorption ionization time-of-flight mass spectrometry mapped the succinylation sites . Most modified lysines are positioned in surface-located loops, numbers 1 and 4 to 7 . Succinylated porin was reconstituted into planar lipid bilayers, and biophysical properties were analyzed and compared to Hib porin: there was an increased average single channel conductance compared to Hib porin (1.24 +/- 0.41 vs . 0.85 +/- 0.40 nanosiemens) . The voltage-gating activity of succinylated porin differed considerably from that of Hib porin . The threshold voltage for gating was decreased from 75 to 40 mV . At 80 mV, steady-state conductance for succinylated porin was 50-55% of the instantaneous conductance . Hib porin at 80 mV showed a decrease to 89-91% of the instantaneous current levels . We propose that surface-located lysine residues are determinants of voltage gating for porin of Haemophilus influenzae type b.

Kansenshogaku Zasshi, 2000 Nov, 74(11), 949 - 53
{Viral infection related to the appearance of acute bacterial respiratory tract infections}; Kobashi Y et al.; To investigate what viruses are related to acute bacterial respiratory tract infections, we prospectively evaluated 113 cases with respiratory tract infections (always accompanying by purulent sputum) experienced between July 1998 and March 2000 . Acute viral infections were detected in 25 cases (22%); 10 cases of influenza A virus and 6 cases of respiratory syncytial (RS) virus . The epidemiology of the influenza A virus and RS virus was mainly identified as from December to February in both winter seasons . A bacteriological examination of sputum cultures isolated 12 cases of Streptococcus pneumoniae and 10 cases of Haemophilus influenzae during the same periods and mixed infections of both viruses and bacteria were recognised in 16 cases (14%) . These results suggest a significantly high percentage of mixed infections of both viruses and bacteria . However, it was unknown whether the patients with acute bacterial respiratory infections had been infected with viruses prior to the bacterial infections . The frequency of appearance of respiratory tract infections tended to increase with the seasonale epidemiology of viral infections.

Infection, 2000 Nov-Dec, 28(6), 351 - 4
Haemophilus influenzae infections among hospitalized adult patients; Furrer M et al.; BACKGROUND: Relatively few data are available about Haemophilus influenzae (Hi) infection among adults . MATERIALS AND METHODS: We studied all adult patients with Hi infection hospitalized between 1988 and 1997 at the University Hospital of Berne . Data were abstracted retrospectively from clinical charts and microbiology records using a standardized questionnaire . RESULTS: 12 invasive and 19 noninvasive Hi infections were observed during the study period.The main clinical manifestations were pneumonia (38.7%), bronchitis (29.0%) and meningitis (12.9%) . Most patients (71.8%) had an underlying condition . Lethality was high (22.6%), especially in pneumonia patients (50%) . The frequency of meningitis caused by Hi serotype b (Hib) seemed to decrease after 1990 when conjugated vaccines against Hib were introduced . CONCLUSION: Hi remains an important cause of lower respiratory and invasive disease associated with high lethality among polymorbid adult patients . The frequency of Hib infections may also decrease in adults due to herd immunity induced by universal vaccination of children.

Int J Antimicrob Agents, 2000 Nov, 16 Suppl 1, S25 - 9
Resistance patterns of lower respiratory tract pathogens in Europe; Marchese A et al.; Resistance to antimicrobial drugs in the major respiratory tract pathogens is known to vary profoundly depending on geographic location . In Europe high rates (>44%) of penicillin-resistance in pneumococci have been recorded in France and Spain, while countries like The Netherlands, the Czech Republic, Austria and Italy are only marginally affected . Similarly, the incidence of macrolide resistance differs widely among European nations with figures ranging from 45.9% (France) to 1.5% (The Netherlands) . Significant percentages (>20%) of co-trimoxazole and doxycycline resistance have been found in France, Spain, Italy, Hungary, Poland and Belgium . The prevailing mechanism of resistance in Haemophilus influenzae is represented by beta-lactamase synthesis for which considerable variations (from 0 to 38.5%) have been evidenced . Ampicillin resistant beta-lactamase negative H . influenzae are very uncommon . Over 90% of Moraxella catarrhalis isolates are beta-lactamase producers without big differences among European countries.

J Clin Microbiol, 2001 Jan, 39(1), 43 - 6
Influence of variations in test methods on susceptibility of Haemophilus influenzae to ampicillin, azithromycin, clarithromycin, and telithromycin; Fuchs PC et al.; The National Committee for Clinical Laboratory Standards standard broth microdilution method for testing the susceptibility of Haemophilus influenzae to ampicillin, azithromycin, clarithromycin, and telithromycin was evaluated by altering one variable at a time . Variables that were tested included age of colony for inoculum preparation, inoculum density, test medium, incubation atmosphere, and incubation time . For the macrolide, azalide, and ketolide agents, incubation in 5 to 7% CO(2) most significantly affected the MICs, producing nearly twofold increases for clarithromycin and telithromycin and a greater than threefold increase for azithromycin . For ampicillin, a 10-fold increase in inoculum density increased the geometric mean MICs for beta-lactamase-negative strains from 1 . 50 to 2.45 microg/ml . In addition, 206 H . influenzae strains were tested for their susceptibilities to the same drugs by the broth microdilution tests in two media, as well as by agar dilution tests, disk diffusion tests, and Etests, on six different agar media . The three standard methods with Haemophilus test medium (HTM) compared favorably with each other except for a high minor discrepancy rate (27%) by the disk diffusion test with ampicillin and clarithromycin . Agar dilution test MICs on the five comparative media were generally higher than those on HTM agar but were only rarely more than one twofold concentration higher . Etest MICs of azithromycin and telithromycin were more than twofold higher than agar dilution and broth microdilution MICs on HTM; ampicillin Etest MICs were nearly twofold lower . The use of media other than HTM agar appears to have a minimal effect on susceptibility test results for the ketolide, azalide, or macrolide drugs that we tested against H . influenzae.

Mol Microbiol, 2001 Jan, 39(2), 341 - 50
Identification of a lipopolysaccharide alpha-2,3-sialyltransferase from Haemophilus influenzae; Hood DW et al.; We have identified a gene for the addition of N-acetylneuraminic acid (Neu5Ac) in an alpha-2,3-linkage to a lactosyl acceptor moiety of the lipopolysaccharide (LPS) of the human pathogen Haemophilus influenzae . The gene is one that was identified previously as a phase-variable gene known as lic3A . Extracts of H . influenzae, as well as recombinant Escherichia coli strains producing Lic3A, demonstrate sialyltransferase activity in assays using synthetic fluorescent acceptors with a terminal galactosyl, lactosyl or N-acetyl-lactosaminyl moiety . In the RM118 strain of H . influenzae, Lic3A activity is modulated by the action of another phase-variable glycosyltransferase, LgtC, which competes for the same lactosyl acceptor moiety . Structural analysis of LPS from a RM118:lgtC mutant and the non-typeable strain 486 using mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy confirmed that the major sialylated species has a sialyl-alpha-(2-3)-lactosyl extension off the distal heptose . This sialylated glycoform was absent in strains containing a lic3A gene disruption . Low amounts of sialylated higher molecular mass glycoforms were present in RM118:lgtC lic3A, indicating the presence of a second sialyltransferase . Lic3A mutants of H . influenzae strains show reduced resistance to the killing effects of normal human serum . Lic3A, encoding an alpha-2,3-sialyltransferase activity, is the first reported phase-variable sialyltransferase gene.

Ir Med J, 2000 Nov, 93(8), 239 - 40
Apparent low immunisation uptake in Dublin: under-performance or under-recording?
Harrington PM, Woodman C, Shannon WF.
Through careful follow-up of a cohort, born March to August 1994, we have recorded the highest ever primary immunisation uptake figures for the Dublin area, with completed uptake for Diphtheria, Tetanus and Oral Polio of 92.1%, Haemophilus influenzae type b of 88.7%, Pertussis of 85.7% and Measles/Mumps/Rubella of 78.1% . Eastern Health Board uptake estimates for the same period are 8.1-21.4% lower . We believe the rigour of our data gathering explains this discrepancy . Evidence is reviewed in support of the hypothesis that Eastern Health Board databases underestimate true immunisation uptake.

Acta Otolaryngol, 2000 Oct, 120(7), 815 - 20
Adaptive bone modeling and remodeling in acute otitis media caused by non-typeable or type B Haemophilus influenzae or Moraxella catarrhalis; Caye-Thomasen P et al.; Experimental studies have shown that acute otitis media caused by Streptococcus pneumoniae alters modeling dynamics in bone tissue structures surrounding the middle ear cavity . Initial resorption of bone is followed by formative activity, seen as massive osteoneogenesis . However, neither resorptive nor formative activity occurs in the otic capsule, supporting the existence of a perilymphatic zone of specialized bone . This study investigates adaptive bone modeling in acute otitis media caused by other bacteria frequently encountered in this disease . Seventy-five rats were inoculated with either non-typeable or type b Haemophilus influenzae, or Moraxella catarrhalis (25 rats in each group) . Five rats from each group were sacrificed on days 4, 8, 16, 60 and 180 post-inoculation . Qualitative as well as quantitative histopathology revealed increasing apposition of new bone on both sides of the original bony wall of the middle ear bulla, i.e . at the inner and outer periosteum . Remodeling activity was seen on later days of sacrifice, as typical osteone (Haversian system) formation . Measured bone thickness in four anatomically well-defined localities progressed to a peak 2 months post-inoculation, followed by some degree of normalization . However, bone thickness was still massively increased 6 months after the acute incident . Except in the otic capsule, resorptive and formative activity was found in all bone tissue structures surrounding the middle ear cavity . These findings were irrespective of the type of inoculated bacteria . However, non-typeable or type b Haemophilus influenzae induces significantly more new bone formation than Moraxella catarrhalis . We conclude that acute otitis media caused by either of the bacteria is accompanied by massive and progressive net osteoneogenesis, already evident on day 4 and peaking 2 months post-inoculation, followed by some degree of normalization . Non-typeable and type b Haemophilus influenzae induce more new bone formation than Moraxella catarrhalis, whereas other features of bone histomorphology were equivalent . The present findings further support the existence of a perilymphatic zone of specialized bone.

Acta Otolaryngol, 2000 Oct, 120(7), 810 - 4
Polyp and fibrous adhesion formation in acute otitis media caused by non-typeable or type b Haemophilus influenzae or Moraxella catarrhalis; Caye-Thomasen P et al.; Among a variety of other histopathologic changes, polyps and fibrous adhesions are readily formed in the middle ear mucosa during experimental acute otitis media (AOM) caused by Streptococcus pneumoniae . Quantitative studies on experimental AOM caused by other bacteria have shown that some of these histopathologic changes, such as adaptive bone modeling and increase in goblet cell density, differ according to the type of bacteria . This investigation surveys polyp and fibrous adhesion formation in experimental AOM caused by either non-typeable or type b Haemophilus influenzae, or Moraxella catarrhalis . Seventy-five rats were inoculated with 1 of these 3 bacteria (25 rats in each of 3 groups) . Five rats from each group were sacrificed on days 4, 8, 16, 60 and 180 post-inoculation . The middle ear mucosae were dissected and histopathologic changes in whole-mount and section preparations were studied using light microscopy . Polyps were found in most ears and in the greatest numbers on the early days; fewer polyps were found on the later days, regardless of the type of bacteria . However, non-typeable and type b H . influenzae induced formation of significantly more polyps than M . catarrhalis . The polyps were primarily located in the epitympanum . Fibrous adhesions were primarily located in the hypotympanum and formed in almost all ears, on all days post-inoculation, regardless of the type of bacteria . Numbers increased to a peak on day 16 and then decreased . Non-typeable and type b H . influenzae induced formation of significantly more adhesions than M . catarrhalis, and the middle ears displayed a higher number of persisting adhesions in the animals inoculated with non-typeable H . influenzae . We conclude that polyps and adhesions are formed in experimental AOM regardless of bacterial type, confirming a pathogenesis based on inflammation . Both types of H . influenzae induce formation of greater numbers of polyps/adhesions than M . catarrhalis, and the non-typeable form causes more adhesive sequelae in the mucosa than the encapsulated type b.

J Chemother, 2000 Oct, 12 Suppl 4, 16 - 20
Resistance in respiratory tract pathogens: an international study 1997-1998; Thornsberry C et al.; Multiple antibiotic resistance threatens current treatment for community-acquired pneumonia (CAP) . This paper presents a summary of resistance data for Streptococcus pneumoniae (6,223 isolates), Haemophilus influenzae (4,016) and Moraxella catarrhalis (1,263) collected from 153 centers throughout Japan, China, UK, Germany, Spain, France, Italy, Brazil and USA . Antiobiotics tested were: beta-lactams (penicillin, ampicillin, co-amoxiclav, cefuroxime, and ceftriaxone), macrolides (azithromycin and clarithromycin), sulphonamide (trimethoprim-sulfamethoxazole), glycopeptide (vancomycin) and fluoroquinolone (levofloxacin) . S . pneumoniae with reduced susceptibility to penicillin were predominant in France, Spain and Japan (54-65%), ,beta-lactamase-producing H . influenzae most common in the USA, France and Spain (>25%) and most M . catarrhalis produced beta-lactamase irrespective of origin . S . pneumoniae susceptibility to azithromycin and clarithromycin varied widely . Levofloxacin was active against almost all isolates in all countries and none was resistant to vancomycin . Because of increasing resistance to older drugs, the newer fluoroquinolones have a role in the therapy of CAP and other respiratory infections, although surveillance studies must continue.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 203 - 7
In vitro and in vivo antibacterial activities of L-084, a novel oral carbapenem, against causative organisms of respiratory tract infections; Miyazaki S et al.; L-084 (a prodrug of LJC 11,036 {L-036}) is a new oral carbapenem . Here we compared the in vitro and in vivo antibacterial activities of L-036 with those of imipenem, faropenem, ceditoren-pivoxil, cefdinir, amoxicillin, and levofloxacin . The MICs at which 90% of the isolates were inhibited of L-036 against methicillin-susceptible staphylococci, Streptococcus pneumoniae including penicillin-resistant organisms, Escherichia coli, Klebsiella pneumoniae, Haemophilus influenzae including ampicillin-resistant organisms, Legionella pneumophila, and Moraxella catarrhalis were equal to or less than 1 microg/ml . In pharmacokinetics studies of L-084 in lungs of mice, the maximum concentration in serum, half-life, and area under the concentration-time curve of this drug were 9.09 microg/g of tissue, 6.18 h, and 31.0 microg . h/ml, respectively . In murine respiratory infection models of penicillin-susceptible and -resistant S . pneumoniae and H . influenzae, the efficacies of L-084 were better than those of reference drugs . Our results indicate that the in vitro high potency and good distribution in the lungs might be the underlying mechanisms of its efficacy in the murine model of pneumonia.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 67 - 72
Susceptibilities of Haemophilus influenzae and Moraxella catarrhalis to ABT-773 compared to their susceptibilities to 11 other agents; Credito KL et al.; The activity of the ketolide ABT-773 against Haemophilus and Moraxella was compared to those of 11 other agents . Against 210 Haemophilus influenzae strains (39.0% beta-lactamase positive), microbroth dilution tests showed that azithromycin and ABT-773 had the lowest MICs (0.5 to 4.0 and 1.0 to 8.0 microg/ml, respectively), followed by clarithromycin and roxithromycin (4.0 to >32.0 microg/ml) . Of the beta-lactams, ceftriaxone had the lowest MICs (</=0.004 to 0.016 microg/ml), followed by cefixime and cefpodoxime (0.008 to 0.125 and </=0.125 to 0.25 microg/ml, respectively), amoxicillin-clavulanate (0.125 to 4.0 microg/ml), and cefuroxime (0 . 25 to 8.0 microg/ml) . Amoxicillin was only active against beta-lactamase-negative strains, and cefprozil had the highest MICs of all oral cephalosporins tested (0.5 to >32.0 microg/ml) . Against 50 Moraxella catarrhalis strains, all of the compounds except amoxicillin and cefprozil were active . Time-kill studies against 10 H . influenzae strains showed that ABT-773, at two times the MIC, was bactericidal against 9 of 10 strains, with 99% killing of all strains at the MIC after 24 h; at 12 h, ABT-773 gave 90% killing of all strains at two times the MIC . At 3 and 6 h, killing by ABT-773 was slower, with 99.9% killing of four strains at two times the MIC after 6 h . Similar results were found for azithromycin, with slightly slower killing by erythromycin, clarithromycin, and roxithromycin, especially at earlier times . beta-Lactams were bactericidal against 8 to 10 strains at two times the MIC after 24 h, with slower killing at earlier time periods . Most compounds gave good killing of five M . catarrhalis strains, with beta-lactams killing more rapidly than other drugs . ABT-773 and azithromycin gave the longest postantibiotic effects (PAEs) of the ketolide-macrolide-azalide group tested (4.4 to >8.0 h), followed by clarithromycin, erythromycin, and roxithromycin . beta-Lactam PAEs were similar and shorter than those of the ketolide-macrolide-azalide group for all strains tested.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 23 - 9
Pharmacodynamics of telithromycin In vitro against respiratory tract pathogens; Odenholt I et al.; Telithromycin (HMR 3647) is a new ketolide that belongs to a new class of semisynthetic 14-membered-ring macrolides which have expanded activity against multidrug-resistant gram-positive bacteria . The aim of the present study was to investigate different basic pharmacodynamic properties of this new compound . The following studies of telithromycin were performed: (i) studies of the rate and extent of killing of respiratory tract pathogens with different susceptibilities to erythromycin and penicillin exposed to a fixed concentration that corresponds to a dose of 800 mg in humans, (ii) studies of the rate and extent of killing of telithromycin at five different concentrations, (iii) studies of the rate and extent of killing of the same pathogens at three different inocula, (iv) studies of the postantibiotic effect and the postantibiotic sub-MIC effect of telithromycin, and (v) determination of the rate and extent of killing of telithromycin in an in vitro kinetic model . In conclusion, telithromycin exerted an extremely fast killing of all strains of Streptococcus pneumoniae both with static concentrations and in the in vitro kinetic model . A slower killing of the strains of Streptococcus pyogenes was noted, with regrowth in the kinetic model of a macrolide-lincosamide-streptogramin B-inducible strain . The strains of Haemophilus influenzae were not killed at all at a concentration of 0.6 mg/liter due to high MICs . A time-dependent killing was seen for all strains . No inoculum effect was seen for the strains of S . pneumoniae, with a 99.9% reduction in the numbers of CFU for all inocula at both 8 h and 24 h . The killing of the strains of S . pyogenes was reduced by 1 log(10) CFU at 8 h and 2 to 3 log(10) CFU at 24 h when the two lower inocula were used but not at all at 8 and 24 h when the highest inoculum was used . For both of the H . influenzae strains there was an inoculum effect, with 1 to 2 log(10) CFU less killing for the inoculum of 10(8) CFU/ml in comparison to that for the inoculum of 10(6) CFU/ml . Overall, telithromycin exhibited long postantibiotic effects and postantibiotic sub-MIC effects for all strains investigated.

Infect Immun, 2001 Jan, 69(1), 307 - 14
Mapping of binding domains of nontypeable Haemophilus influenzae HMW1 and HMW2 adhesins; Dawid S et al.; Nontypeable Haemophilus influenzae is an important cause of localized respiratory tract disease, which begins with colonization of the upper respiratory mucosa . In previous work we reported that the nontypeable H . influenzae HMW1 and HMW2 proteins are high-molecular-weight nonpilus adhesins responsible for attachment to human epithelial cells, an essential step in the process of colonization . Interestingly, although HMW1 and HMW2 share significant sequence similarity, they display distinct cellular binding specificities . In order to map the HMW1 and HMW2 binding domains, we generated a series of complementary HMW1-HMW2 chimeric proteins and examined the ability of these proteins to promote in vitro adherence by Escherichia coli DH5alpha . Using this approach, we localized the HMW1 and HMW2 binding domains to an approximately 360-amino-acid region near the N terminus of the mature HMW1 and HMW2 proteins . Experiments with maltose-binding protein fusion proteins containing segments of either HMW1 or HMW2 confirmed these results and suggested that the fully functional binding domains may be conformational structures that require relatively long stretches of sequence . Of note, the HMW1 and HMW2 binding domains correspond to areas of maximal sequence dissimilarity, suggesting that selective advantage associated with broader adhesive potential has been a major driving force during H . influenzae evolution . These findings should facilitate efforts to develop a subcomponent vaccine effective against nontypeable H . influenzae disease.

Infect Immun, 2001 Jan, 69(1), 221 - 7
Haemophilus influenzae porin contributes to signaling of the inflammatory cascade in rat brain; Galdiero M et al.; In the present study we observed that the Haemophilus influenzae type b (Hib) porin, among the different surface bacterial components, is involved in the pathophysiology of bacterial meningitis . This study demonstrates that inoculation of Hib porin into the fourth cerebral ventricle causes the simultaneous expression of interleukin-1alpha (IL-1alpha), tumor necrosis factor alpha (TNF-alpha), and macrophage inflammatory protein 2 (MIP-2) at 6 h after inoculation . At 24 h, the expression of MIP-2 decreases while the expression of IL-1alpha and TNF-alpha increases . The mRNA expression of IL-1alpha, TNF-alpha, and MIP-2 is correlated with injury to the blood-brain barrier as demonstrated by the appearance of serum proteins and leukocytes in cerebrospinal fluid and by the increase in brain water content.

Infect Immun, 2001 Jan, 69(1), 154 - 8
Cloning and expression of the Actinobacillus actinomycetemcomitans thioredoxin (trx) gene and assessment of cytokine inhibitory activity; Henderson B et al.; Thioredoxin is a ubiquitous redox control and cell stress protein . Unexpectedly, in recent years, thioredoxins have been found to exhibit both cytokine and chemokine activities, and there is increasing evidence that this class of protein plays a role in the pathogenesis of inflammatory diseases . In spite of this evidence, it has been reported that the oral bacterium and periodontopathogen Actinobacillus actinomycetemcomitans secretes an immunosuppressive factor (termed suppressive factor 1 {SF1} {T . Kurita-Ochiai and K . Ochiai, Infect . Immun . 64:50-54, 1996}) whose N-terminal sequence, we have determined, identifies it as thioredoxin . We have cloned and expressed the gene encoding the thioredoxin of A . actinomycetemcomitans and have purified the protein to homogeneity . The A . actinomycetemcomitans trx gene has 52 and 76% identities, respectively, to the trx genes of Escherichia coli and Haemophilus influenzae . Enzymatic analysis revealed that the recombinant protein had the expected redox activity . When the recombinant thioredoxin was tested for its capacity to inhibit the production of cytokines by human peripheral blood mononuclear cells, it showed no significant inhibitory capacity . We therefore conclude that the thioredoxin of A . actinomycetemcomitans does not act as an immunosuppressive factor, at least with human leukocytes in cultures, and that the identity of SF1 remains to be elucidated.

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