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