J Clin Microbiol, 1995 Jun, 33(6), 1486 - 91 Rapid bacterial antigen detection is not clinically useful; Perkins MD et al.; Latex agglutination (LA) of capsular polysaccharide bacterial antigen is a frequently performed laboratory procedure, but its use is controversial . To assess the clinical utility of this test, we reviewed all LA tests performed over a 10-month period at two sites, a major university-based referral center and a private specialty pediatric hospital . Samples were assayed either individually or as a panel for the group B streptococcus, Streptococcus pneumoniae, Haemophilus influenzae, and three sets of Neisseria meningitidis serogroups (A and Y, C and W135, and B and Escherichia coli K1) . Of 5,169 assays performed on 1,268 clinical samples (786 urine and 478 cerebrospinal fluid, 3 pleural fluid, and 1 synovial fluid sample), 57 (1.1%) were positive, including 1.7% of urine and 0.3% of cerebrospinal fluid samples . All LA true-positive cerebrospinal fluid samples showed the causative microorganisms by Gram stain . Detailed chart review of these 57 positive samples showed that the LA result was false-positive in 31 (54%), true-positive in 22 (38%), and indeterminate in 4 (7%) samples . Therapy was not altered on the basis of any of the true-positive LA results . The 31 false-positive results led to additional cost, prolonged hospitalization, and some clinical complications . Total patient charges were $175,000 ($7,954 per true-positive), with no detectable clinical benefit . Our retrospective study does not support the current use of LA for rapid antigen detection . What, if any, specific indications exist for this test remain to be elucidated. J Fla Med Assoc, 1995 Jun, 82(6), 401 - 2 Cost-effectiveness of Haemophilus influenzae type b conjugate vaccine program in Florida; Midani S et al.; Introduction of Haemophilus influenzae type b (Hib) vaccine has significantly decreased this disease's incidence in childhood . The cost-effectiveness and economic impact of the Hib immunization program in Florida were investigated and three periods compared: I = 1984-1988; II = 1989-1990; and III = 1991-1992 . The cost per year of Hib disease in Period I was $27.48 million while that in Periods II and III was $15.95 million and $.88 million respectively . The total savings in millions were: Periods I and II = $11.53; Periods II and III = $11.07; and Periods I and III = $22.6 . The greatest saving was realized between Periods I and III because of the initiation of immunization with the Hib vaccine starting at two months of age during Period III . Hib immunization is cost-effective and significant savings would more than pay for the cost of the program. Drugs, 1995 Jun, 49(6), 1007 - 22 Cefixime . A review of its therapeutic efficacy in lower respiratory tract infections; Markham A et al.; Cefixime is an orally active third generation cephalosporin with in vitro antibacterial activity against most important lower respiratory pathogens . The drug is active against Haemophilus influenzae, Moraxella catarrhalis and penicillin-susceptible Streptococcus pneumoniae but not Staphylococcus aureus . Cefixime has a long elimination half-life (3 hours compared with 0.5 hours for cefaclor and 1.5 hours for cefalexin), which allows once daily administration . Several trials have established the clinical efficacy of the drug in patients with lower respiratory tract infection (LRTI) . In comparative studies cefixime had similar efficacy to amoxicillin +/- clavulanic acid, cefaclor, cefalexin, cefuroxime axetil and clarithromycin . Trials evaluating the efficacy of cefixime as the oral component of intravenous to oral switch therapy have produced promising preliminary results although further carefully designed trials are needed in this area . As with certain other drugs of its class, gastrointestinal disturbances are the most frequently reported adverse events in patients taking cefixime and cases of pseudomembranous colitis have been reported . Thus, cefixime is an effective treatment for mild to moderate LRTI and may have a role as the oral component of intravenous to oral switch therapy although further well designed studies are needed to confirm initial favourable results in this important emerging area of antibacterial therapy. Immunol Cell Biol, 1995 Jun, 73(3), 258 - 65 Protection against non-typable Haemophilus influenzae following sensitization of gut associated lymphoid tissue: role of specific antibody and phagocytes; Wallace FJ et al.; Rats intestinally immunized with whole killed non-typable Haemophilus influenzae clear this organism from the lungs faster than non-immunized rats . This study investigated the role of antibody and phagocytes in the clearance mechanism . First, dose-response experiments demonstrated that while lowering the dose of non-typable H . influenzae reduced the level of detectable specific antibody in bronchial washings, the ability to accelerate bacterial clearance persisted to much lower doses . Second, specificity experiments showed that intestinal immunization with non-typable H . influenzae cross-protected against Pseudomonas aeruginosa, even though antibodies were not absorbed out of serum by incubation with P . aeruginosa . Third, serum antibody was shown to be bactericidal for non-typable H . influenzae in the presence of complement (P < 0.05), while bronchial washings antibody was not . The bactericidal effect of the serum was abrogated by the addition of bronchial washings . Fourth, an ELISA quenching assay demonstrated that neutrophils from intestinally immunized rats were able to phagocytose more bacteria in a given time period (P < 0.05) than unimmunized rats and rats immunized by other routes . In the fifth experiment, the chemotactic response of neutrophils to casein was shown to be significantly depressed by the addition of bronchial washings obtained from immunized rats (P < 0.01) . It is proposed that specific antibody in bronchial washings does not have a direct role in opsonizing bacteria for killing or phagocytosis, but instead has an anti-inflammatory effect . Non-specific effectors such as neutrophils driven by specific immune cells are a likely means of clearance of bacteria following intestinal immunization and acute challenge. Fam Pract, 1995 Jun, 12(2), 155 - 8 The high vaginal swab in general practice: clinical correlates of possible pathogens; Dykhuizen RS et al.; Clinical features, diagnosis and treatment of 286 women whose high vaginal swabs (HVS) submitted by their general practitioners showed pure, heavy growth of Staphylococcus aureus, beta haemolytic streptococci groups A, C or G, Streptococcus milleri, Streptococcus pneumoniae or Haemophilus influenzae were analysed . Women with group A, C and G streptococci frequently had clinical vulvovaginitis and although the numbers were too small for statistical confirmation, S . pneumoniae and H . influenzae appeared to cause clinical disease as well . The association of S . aureus or S . milleri with clinical vulvovaginitis was much less convincing . It seems relevant for laboratories to report sensitivities for group A, C and G streptococci . Further research is needed to determine the pathogenicity of S . pneumoniae and H . influenzae. Eur Respir J, 1995 Jun, 8(6), 948 - 53 The isolation and characterization of non-typeable Haemophilus influenzae from the sputum of adult cystic fibrosis patients; Bilton D et al.; The role of non-typeable Haemophilus influenzae in cystic fibrosis (CF) remains unclear . We wanted, therefore, to determine the presence and characteristics of non-typeable H . influenzae in sputum samples from patients with CF . In order to do this, we have assessed sputum samples from 55 consecutive clinically stable patients seen routinely at an adult CF out-patient clinic . Quantitative bacterial culture was performed using a selective media containing cefsoludin, and isolates were characterized by biotyping and outer membrane protein profile analysis . In 17 (30%) of these samples, non-typeable H . influenzae was isolated and was present in similar viable numbers (mean 7.7 x 10(8) colony-forming units (cfu).mL-1; SEM 3.1) to Pseudomonas aeruginosa (mean 8 x 10(8) cfu.mL-1: SEM 2.4) . All non-typeable H . influenzae isolates recovered were beta-lactamase negative and sensitive to a range of antibiotics . Several biotypes and outer membrane protein profiles were observed, with no apparent association between these two phenotypic characteristics . The study showed that large numbers of non-typeable H . influenzae are often present in sputum from adult patients with CF . Further longitudinal studies of outer-membrane protein profile analysis are required to determine the dynamics of non-typeable H . influenzae colonization in individual patients and the clinical significance. Allergy, 1995 Jun, 50(6), 528 - 31 Characterization of the antibody response to a Haemophilus influenzae type b conjugate vaccine in children with recurrent lower respiratory tract infection; Kristensen K et al.; Children with recurrent lower respiratory tract infection (RLRI) may respond poorly to polysaccharide antigens . To examine how such children respond to a polysaccharide coupled to a protein carrier, we immunized 15 children with RLRI aged 8-69 months and 15 carefully age-matched healthy controls once with a Haemophilus influenzae type b (Hib) conjugate vaccine . Total IgG subclasses, total antipolysaccharide Hib antibodies, and antipolysaccharide Hib antibodies of IgM, IgG, IgA, and IgG1-4 specificity were determined by ELISA . There were no significant differences between the two groups in any single total IgG subclass, but total IgG measured as the sum of all four subclasses was significantly lower in the children with RLRI than in the controls (P = 0.036) . Before vaccination, the children with RLRI had significantly less IgG antipolysaccharide Hib antibody than the controls (P = 0.005), whereas 1 month later they had significantly more IgM antibody (P = 0.038) . No other significant differences were found between the groups before or after immunization with respect to antipolysaccharide Hib antibodies . Since naturally occurring IgG antibodies are thought to be acquired partly as a consequence of antigenic stimulation on mucosal surfaces, we hypothesize that the low level of specific IgG found before immunization, as well as the low total IgG in the children with RLRI, may reflect an impaired ability to prime through mucosal surfaces . This is supported by our finding of an increased IgM response to Hib conjugate vaccine in these children, since this isotype predominates in the primary immune response, i.e., in the absence of immunologic memory.(ABSTRACT TRUNCATED AT 250 WORDS) Oral Microbiol Immunol, 1995 Jun, 10(3), 151 - 9 Molecular and immunological characterization of a 64-kDa protein of Actinobacillus actinomycetemcomitans; Nakano Y et al.; The 64-kDa protein to which about half the sera from patients with localized juvenile periodontitis and rapidly progressive periodontitis reacted strongly was purified from Actinobacillus actinomycetemcomitans Y4 . Determination of the N-terminal sequence of the protein revealed that it was a GroEL-like protein . The DNA fragment containing the groEL gene of A . actinomycetemcomitans was amplified by polymerase chain reaction, and the groESL operon was cloned by using colony hybridization with the amplified fragment from A . actinomycetemcomitans chromosomal DNA . Sequence analysis revealed that structures of the operon and its products were typical in gram-negative bacteria . Rabbit polyclonal antibodies to the 64-kDa protein cross-reacted with approximately 65-kDa proteins of Haemophilus aphrophilus, Haemophilus influenzae, Haemophilus paraphrophilus, Escherichia coli and Eikenella corrodens but not with any cellular proteins of Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum . It is possible that antibodies reactive to the 64-kDa protein in periodontitis patients are induced by the cross-reactivity with the hsp60 proteins of other bacteria. J Hosp Infect, 1995 Jun, 30 Suppl, 313 - 21 Use of conjugate vaccines to prevent meningitis caused by Haemophilus influenzae type b or Streptococcus pneumoniae; Eskola J; Antibodies to the capsular polysaccharides of Haemophilus influenzae type b or Streptococcus pneumoniae protect against meningitis caused by these bacteria . Many of the polysaccharides are poorly immunogenic, especially in infants, but can be turned to highly immunogenic vaccines by covalent conjugation to a protein carrier . On the basis of the good protection observed in several trials, H . influenzae type b conjugates have been accepted for wide use . This experience has also provided direction for the development of new conjugates against infections caused by the most common serotypes of S . pneumoniae . First results from immunogenicity studies of these pneumococcal conjugate vaccines are promising. Clin Otolaryngol, 1995 Jun, 20(3), 219 - 23 The microbiology and antibiotic treatment of peritonsillar abscesses; Prior A et al.; Pus from 53 peritonsillar abscesses was cultured and associations between the microbiological results and clinical data were investigated with the aim of developing a clinical protocol for treatment . A positive culture grew in 85% of quinsies and of these 16% produced aerobes and 84% anaerobes . Penicillin-resistant organisms were grown from 32% of patients and all but one of these organisms (Haemophilus influenzae) was sensitive to metronidazole . There was no association between clinical presentation and cultured organism which could guide treatment, hence we recommend penicillin and metronidazole as the antibiotic regimen of choice in the treatment of peritonsillar abscesses because of its effectiveness in 98% of patients. J Formos Med Assoc, 1995 Jun, 94(6), 351 - 4 Serotypes, biotypes and antibiotic susceptibility of 126 clinical isolates of Haemophilus influenzae; Chiu CH et al.; Serotypes, biotypes, and antibiotic susceptibility of 126 Haemophilus influenzae isolates were determined . Five of the 126 isolates were from blood and were encapsulated type b strains; those taken from other sites were not typable . There were 13% biotype I, 36% biotype II, 38% biotype III, 5% biotype IV, 4% biotype V, and 4% biotype VI isolates . Antibiotic susceptibility tests using the standard disk diffusion method showed the following resistance: ampicillin 51%, cefamandole 10%, cefuroxime 3%, chloramphenicol 28%, tetracycline 37% and sulfamethoxazole-trimethoprim 49% . None of the five type b isolates were resistant to cefotaxime, a third generation cephalosporin . The second generation cephalosporins, cefamandole and cefuroxime, showed a superior activity against H . influenzae isolates, compared to other antibiotics . Multiple drug resistance was found in 64 (51%) isolates . Four of the five type b isolates were resistant to multiple drugs . The multiple-resistance pattern most frequently observed was to ampicillin, chloramphenicol, tetracycline and sulfamethoxazole-trimethoprim . Most clinical isolates did not contain plasmids; therefore, the antibiotic resistance of these H . influenzae strains was probably chromosome-mediated. East Afr Med J, 1995 Jun, 72(6), 376 - 8 Analysis of the results of routine lumbar puncture after a first febrile convulsion in Hofuf, Al-Hassa, Saudi Arabia; Laditan AA; Cerebrospinal fluid (CSF) was analysed to determine a lumbar puncture (LP) yield for meningitis in 95 children who presented with their first febrile convulsions between July 1993 and June 1994 . There were 52 males and 43 females aged six months to six years with a mean age of 21.9 +/- 13.0 months at presentation . 87(91.6%) had simple febrile convulsions (SFC) while the remaining 8(8.4%) had complex febrile convulsions (CFC) . The majority of the subjects presented with a sudden onset of convulsions that were preceded by a day or two history of fever, coryza, cough and respiratory distress while others had their convulsions preceded by fever and passage of bloody stools . The LP yield for meningitis in this series was 6.3% . The CSF analysis revealed six cases of meningitis comprising an eight month old infant with Haemophilus influenzae type B (HIB) meningitis, two partially treated pyogenic meningitis and three aseptic meningitis . All of them had presented with febrile convulsions without signs of meningeal irritation . Excluding aseptic meningitis from this series, a 3.1% LP yield for pyogenic meningitis is significant enough to recommend continued performance of LP in children with first febrile convulsions, especially if under the age of eighteen months. Ann N Y Acad Sci, 1995 May 31, 754, 289 - 99 Defining surrogate serologic tests with respect to predicting protective vaccine efficacy: poliovirus vaccination; Sutter RW et al.; Inactivated and trivalent oral poliovirus vaccines contain either formalin-inactivated or live, attenuated poliovirus, respectively, of the three serotypes . Interference among the three attenuated poliovirus serotypes was minimized with a "balanced-formulation" vaccine, and serologic responses after IPV were optimized by adjusting the antigenic content of each inactivated poliovirus serotype . Seroconversion is dependent on both the relative content as well as the absolute quantity of virus in the vaccine . The "gold standard" method to assess humoral antibody responses following vaccination is the neutralization assay . Any detectable titer of neutralizing antibody against poliovirus is considered protective against clinical paralytic diseases . Recently, standard procedures were adopted for conducting neutralization assays . Efforts are being undertaken now to develop a combined diphtheria and tetanus toxoids and pertussis vaccine and IPV vaccine in the United States using a dual-chambered syringe that mixes the content of both vaccines at the time of injection; this approach is necessary to overcome the potential detrimental effect of thimerosal on IPV (the preservative in DTP) . Other vaccines that combine DTP and/or Haemophilus influenzae type b and/or hepatitis B with IPV appear feasible but require further investigation . New combination vaccines should induce similar or superior levels of neutralizing antibody in serum for individual protection against paralytic disease and mucosal immunity that effectively decreases viral replication in the intestine and pharynx for population protection against transmission of poliovirus. Ann N Y Acad Sci, 1995 May 31, 754, 278 - 88 Laboratory correlates of protection against Haemophilus influenzae type b disease . Importance of assessment of antibody avidity and immunologic memory; Granoff DM et al.; The concentration of serum antibody to the Haemophilus influenzae type b polysaccharide sufficient to confer protection against Hib disease has been estimated to range from 0.15 to 1.0 microgram/ml as measured by conventional antigen binding assays . However, the ability of these serologic tests to predict vaccine equivalence and/or protective efficacy is limited since there are important qualitative differences in vaccine-induced anti-PRP antibody, such as isotype, variable region usage, and antibody avidity . These differences may profoundly affect the biologic activity of the antibody . Also, Hib conjugate vaccination primes infants for memory antibody responses to a subsequent encounter with PRP, and immunologic priming can occur in infants with very low serum anti-PRP antibody responses to conjugate vaccination, or in those whose antibody concentrations have declined after vaccination . Primed infants are likely to be protected against Hib disease in the absence of "protective" serum antibody concentrations because priming permits a rapid serum anti-PRP antibody response upon encountering the organism . Thus, quantitative assessment of immunogenicity, by itself, is insufficient to predict vaccine equivalence or protective efficacy . In defining surrogate serologic tests for prediction of vaccine efficacy, assessments of antibody avidity and induction of immunologic memory should be included . Ideally, these assessments should be supplemented with antibody functional assays such as complement-mediated bactericidal activity, opsonic activity, or passive protection in animal models of disease. Ann N Y Acad Sci, 1995 May 31, 754, 108 - 13 Combination vaccines for diphtheria, tetanus, pertussis, and Haemophilus influenzae type b; Paradiso PR; The ability to combine the standard DTP and Haemophilus b conjugate vaccine considerably simplifies the childhood immunization schedule and process . In addition to reducing the number of immunizations by half, the combination product reduces administrative aspects associated with vaccination including tracking . Simplification of the immunization process should have a positive impact on the vaccine delivery and utilization . Perhaps more importantly, an ability to create combination vaccines will be critical for inclusion of new antigens appropriate for infant vaccines . The combination of DTP and HbOC reduces the number of immunizations routinely given at 2, 4, and 6 months of age by half . Since it is unlikely that parents or pediatricians will accept more than two shots per visit, this reduction is critical . As new vaccines are licensed for such important childhood pathogens as Streptococcus pneumoniae and respiratory syncytial virus, designing stable combination products will become even more critical . Having stated that, we must also not lose site of the fact that combination products must meet the criteria for stability, safety, and efficacy comparable to the separately delivered products . These considerations are not trivial . In the development of Tetramune (DTP-HbOC), stability of the product and consistency of the immune response were critical design parameters for both the preclinical and clinical research . Likewise, the experience with other DTP-Haemophilus b combinations has shown that simple mixing of products prior to injection can reduce the immune response in ways that are not necessarily predictable . In contrast, the response to each of the components of Tetramune was in fact higher than when the vaccines were given separately . This increased response to all of the antigens was not anticipated based on the vaccine composition and points to the need for not only physical characterization of new combinations, but also clinical testing of final combined products before they are introduced for routine use. JAMA, 1995 May 24-31, 273(20), 1598 - 604 Molecular analysis of bacterial pathogens in otitis media with effusion; Post JC et al.; OBJECTIVE--To determine if the polymerase chain reaction (PCR) can detect bacterial DNA in pediatric middle ear effusions that are sterile by standard cultural methods . DESIGN--Single-center, blinded, comparative study of diagnostic assays . The PCR-based detection systems for Moraxella catarrhalis, Haemophilus influenzae, and Streptococcus pneumoniae were designed and validated using a battery of DNAs obtained from cultured bacteria . Chronic middle ear effusion specimens were collected and comparatively analyzed by culture and the PCR . SETTING--Tertiary care pediatric hospital . PATIENTS--A total of 97 middle ear effusions were collected from pediatric outpatients at Children's Hospital of Pittsburgh (Pa) during myringotomy and tube placement for chronic otitis media with effusion (duration > 3 months) . All patients had failed multiple courses of antimicrobial therapy and were diagnosed by a combination of validated otoscopy and tympanograms . MAIN OUTCOME MEASURE--Differences in the percentage of positive test results between PCR-based assays and culture for M catarrhalis, H influenzae, and S pneumoniae . RESULTS--Of the 97 specimens of otitis media with effusion, 28 (28.9%) tested positive by both culture and PCR for M catarrhalis, H influenzae, or S pneumoniae . An additional 47 specimens (48%) were PCR positive/culture negative for these three bacterial species . Thus, 75 (77.3%) of the 97 specimens tested PCR positive for one or more of the three test organisms . The minimum number of bacterial genomic equivalents present in the average culture-negative ear was estimated to be greater than 10(4) based on dilutional experiments . CONCLUSIONS--The PCR-based assay systems can detect the presence of bacterial DNA in a significant percentage of culturally sterile middle ear effusions . While this finding is not proof of an active bacterial infectious process, the large number of bacterial genomic equivalents present in the ears is suggestive of an active process. Tidsskr Nor Laegeforen, 1995 May 20, 115(13), 1604 - 6 {Vaccine against Haemophilus influenzae type b--antibody response and adverse effects}; Nokleby H et al.; In 1992 it was decided to implement vaccination against Haemophilus influenzae type b (Hib) in the Norwegian vaccination programme . The chosen vaccine consists of Hib-polysaccharide conjugated to tetanus toxoid . To suit the Norwegian vaccination schedule, Hib-vaccine was given together with vaccine against diphtheria, tetanus and pertussis (DTP) at three, five and ten months of age . Since all earlier studies with the Hib-vaccine have used other time schedules, a pilot study was conducted to test antibody response and side effects when using the Norwegian schedule . 44 infants were vaccinated with Hib-vaccine and DTP at three, five and ten months of age . Blood samples showed that 95% had protective antibody titer against Hib after two doses, 100% after three . The response to the other antigens was adequate . There were only minor side effects . After introduction of the Hib-vaccine the incidence of invasive Hib-infection in children below three years of age has decreased considerably. J Chemother, 1995 May, 7 Suppl 1, 16 - 20 Amoxicillin/clavulanic acid vs cefetamet pivoxil in the treatment of acute exacerbation of chronic bronchitis (AECB) in adults; Behler PG et al.; In this open comparative and prospective study 180 adults of either sex were randomised to treatment with either amoxicillin/clavulanic acid (AMC) 500/125mg tid or cefetamet pivoxil (CAT) 500mg bid for 7 days . Demographic data and assessable findings were similar in both groups . Clinical outcomes of 169 assessable patients showed high efficacy of both drugs: 92% with AMC and 96% with CAT . Bacteriological response rates were equivalent in 141 evaluable cases: 84% vs . 89%, respectively . Baseline susceptibility testing (DIN) revealed a notable number of Haemophilus species either intermediately susceptible or resistant to AMC . Gastrointestinal disorders predominated among the adverse events with diarrhea occurring nearly twice as often in the AMC group . CAT is an effective and safe alternative option in the treatment of AECB in adults . The advantage of CAT is its enhanced activity against gram-negative bacteria . It is well tolerated. Rev Inst Med Trop Sao Paulo, 1995 May-Jun, 37(3), 257 - 60 Bacterial antigen detection in cerebrospinal fluid by the latex agglutination test; Landgraf IM et al.; Eighty purulent cerebrospinal fluid (CSF) samples from patients with clinical evidence of meningitis were studied using the Directigen latex agglutination (LA) kit to determine the presence of bacterial antigen in CSF . The results showed a better diagnostic performance of the LA test than bacterioscopy by Gram stain, culture and counterimmunoelectrophoresis (CIE), as far as Neisseria meningitidis groups B and C, and Haemophilus influenzae type b are concerned, and a better performance than bacterioscopy and culture considering Streptococcus pneumoniae . Comparison of the results with those of culture showed that the LA test had the highest sensitivity for the Neisseria meningitidis group C . Comparing the results with those of CIE, the highest levels of sensitivity were detected for N . meningitidis groups B and C . Regarding specificity, fair values were obtained for all organisms tested . The degree of K agreement when the LA test was compared with CIE exhibited better K indices of agreement for N . meningitidis groups B and C. J Pediatr, 1995 May, 126(5 Pt 1), 799 - 806 Bacteriologic failure of amoxicillin-clavulanate in treatment of acute otitis media caused by nontypeable Haemophilus influenzae; Patel JA et al.; OBJECTIVE: To evaluate the rate of bacteriologic failure of amoxicillin-clavulanate in the treatment of acute otitis media (AOM) and to identify the risk factors associated with failure . METHODS: Ninety-nine subjects (mean age, 21.4 months) with AOM were treated with amoxicillin-clavulanate in two prospective study trials that compared efficacy of two experimental antibiotics with amoxicillin-clavulanate . Tympanocentesis for microbiologic studies was performed in all subjects at enrollment; at 3 to 6 days, during amoxicillin-clavulanate therapy; and at other times when clinically indicated . The subjects were followed up for 1 month . Clinical, bacteriologic, and virologic characteristics of the subjects were analyzed . RESULTS: Bacteriologic failure of treatment occurred in none of 39 subjects (0%) with Streptococcus pneumoniae, two of 25 (8%) with Moraxella catarrhalis, and 11 of 29 (38%) with nontypeable Haemophilus influenzae (NTHi) infection . The failure rate for NTHi was higher than that for other pathogens (p = 0.0007) and was increased when compared with the preceding study period (p = 0.017) . Bacteriologic failure was also associated with clinical failure (p = 0.041) . In subjects with AOM caused by NTHi the rates of adequate drug compliance were comparable in both success and failure groups . Antimicrobial susceptibility testing by minimum inhibitory concentration and minimum bactericidal concentration (MIC/MBC) assays showed that amoxicillin-clavulanate resistance was not significantly associated with bacteriologic failure of treatment . However, in two subjects, MIC/MBC of the NTHi isolates during therapy were higher than MIC/MBC of the isolates before therapy; these strains of isolates pretherapy and during therapy were discordant as determined by outer membrane protein analysis . The bacteriologic failure rate was higher in nonwhite boys (p = 0.026) and in subjects with a history of three or more previous episodes of AOM (p = 0.008) . Other factors such as age, bilaterality of disease, polymicrobial infection, and biotype pattern of NTHi were not associated with treatment failure . When children with adequate drug compliance were analyzed separately, only those with concomitant viral infection of the nasopharynx or middle ear were found to be at an increased risk of bacteriologic failure of treatment (p = 0.04) . CONCLUSIONS: The bacteriologic failure rate of amoxicillin-clavulanate therapy for AOM caused by NTHi was higher in the current study period than in the preceding period . Factors contributing to treatment failure were race, gender, proneness to otitis, and concomitant viral infection. J Infect Dis, 1995 May, 171(5), 1217 - 22 Response of recent human immunodeficiency virus seroconverters to the pneumococcal polysaccharide vaccine and Haemophilus influenzae type b conjugate vaccine; Weiss PJ et al.; Antibody responses in recent human immunodeficiency virus (HIV) seroconverters to 2 vaccines were studied . Twenty patients infected with HIV for < 18 months and 15 HIV-seronegative controls were vaccinated with the 23-valent pure polysaccharide pneumococcal vaccine and the Haemophilus influenzae type b (Hib) capsular polysaccharide diphtheria CRM197 protein toxoid conjugate vaccine in separate arms . Despite increased levels of total serum IgG, recent seroconverters and controls showed similar specific IgG responses for 6 of 7 antigens . Baseline levels were equivalent in both groups, as were peak levels of IgG at 1 month to conjugated polysaccharide (Hib), delayed-type hypersensitivity, and pneumococcal capsular serotypes 4, 6B, 12F, and 14 . At 6 months, IgG levels were similar for 4 of 7 antigens . Antibody responses to pure pneumococcal capsular polysaccharides and to a protein recall antigen were most often similar among recent seroconverters and seronegative controls . Both total levels and fold-rises of IgG to the Hib conjugate were similar in the 2 groups . Immunization of HIV-infected patients soon after seroconversion rather than later appears to improve antibody responses. J Bacteriol, 1995 May, 177(10), 2950 - 2 HpaII methyltransferase is mutagenic in Escherichia coli; Bandaru B et al.; A genetic reversion assay to study C-to-T mutations within CG sites in DNA is described . It was used to demonstrate that the presence of HpaII methyltransferase (MTase) in Escherichia coli causes a substantial increase in C-to-T mutations at CG sites . This is similar to the known mutagenic effects of E . coli MTase Dcm within its own recognition sequence . With this genetic system, a homolog of an E . coli DNA repair gene in Haemophilus parainfluenzae was tested for antimutagenic activity . Unexpectedly, the homolog was found to have little effect on the reversion frequency . The system was also used to show that HpaII and SssI MTases can convert cytosine to uracil in vitro . These studies define 5-methylcytosine as an intrinsic mutagen and further elaborate the mutagenic potential of cytosine MTases. J Bacteriol, 1995 May, 177(10), 2644 - 53 A gene cluster involved in the utilization of both free heme and heme:hemopexin by Haemophilus influenzae type b; Cope LD et al.; The utilization of heme bound to the serum glycoprotein hemopexin by Haemophilus influenzae type b (Hib) strain DL42 requires the presence of the 100-kDa heme:hemopexin-binding protein encoded by the hxuA gene (M . S . Hanson, S . E . Pelzel, J . Latimer, U . Muller-Eberhard, and E . J . Hansen, Proc . Natl . Acad . Sci . USA 89:1973-1977, 1992) . Nucleotide sequence analysis of a 5-kb region immediately upstream from the hxuA gene revealed the presence of two genes, designated hxuC and hxuB, which encoded outer membrane proteins . The 78-kDa HxuC protein had similarity to TonB-dependent outer membrane proteins of other organisms, whereas the 60-kDa HxuB molecule most closely resembled the ShlB protein of Serratia marcescens . A set of three isogenic Hib mutants with cat cartridges inserted individually into their hxuA, hxuB, and hxuC genes was constructed . None of these mutants could utilize heme:hemopexin . The hxuC mutant was also unable to utilize low levels of free heme, whereas both the hxuA and hxuB mutants could utilize free heme . When the wild-type hxuC gene was present in trans, the hxuC mutant regained its ability to utilize low levels of free heme but still could not utilize heme:hemopexin . The hxuA mutant could utilize heme:hemopexin when a functional hxuA gene from a nontypeable H . influenzae strain was present in trans . Complementation analysis using this cloned nontypeable H . influenzae hxuA gene also indicated that the HxuB protein likely functions in the release of soluble HxuA from the Hib cell . These studies indicate that at least two and possible three gene products are required for utilization of heme bound to hemopexin by Hib strain DL42. Arch Pediatr Adolesc Med, 1995 May, 149(5), 537 - 40 Epidemiology, etiology, and clinical features of septic arthritis in children younger than 24 months; Yagupsky P et al.; OBJECTIVE: To examine the incidence, etiology, and clinical features of septic arthritis in patients younger than 24 months . DESIGN: Retrospective, 1988 through 1993 period, chart review-based survey . PATIENTS: All children with bacteriologically proved septic arthritis that was diagnosed at a medical center serving southern Israel (population 320,000) . Septic arthritis was defined by clinical evidence of joint inflammation and a positive synovial fluid or blood culture, antigen detection test, or a standard tube agglutination titer of 160 or greater for Brucella species . INTERVENTIONS: None . RESULTS: During the 6-year period, 40 children had septic arthritis diagnosed . Twenty-six (65%) were male . The annual incidence of septic arthritis was 37.1 per 100,000 . The two most common organisms isolated were Kingella kingae in 19 (48%) and Haemophilus influenzae type b in eight (20%) . The clinical presentation was frequently mild: a body temperature of less than 38.3 degrees C was recorded in 14 (35%) of 40 children, leukocyte count of less than 15 x 10(9)/L in 13 (34%) of 38, and erythrocyte sedimentation rate of less than 30 mm per hour in four (11%) of 35 . In eight (36%) of 22 patients, less than 50 x 10(9)/L leukocytes were counted in the synovial fluid . CONCLUSIONS: The diagnosis of septic arthritis in young children requires a high index of suspicion, and the disease cannot be excluded on the basis of lack of fever or normal results of laboratory tests . Kingella kingae appears to be the most common cause of septic arthritis in patients younger than 24 months, although confirmatory studies from other geographic areas are still needed. Infect Immun, 1995 May, 63(5), 2082 - 6 Molecular cloning of Proteus mirabilis uroepithelial cell adherence (uca) genes; Cook SW et al.; Proteus mirabilis bacteria are a common cause of hospital-acquired urinary tract infection . In a previous study, we described a P . mirabilis fimbrial protein, UCA, that adhered to human uroepithelial cells . Genes sufficient for expression of UCA adherence were cloned into Escherichia coli K-12 . E . coli bacteria that contained the uca recombinant plasmid adhered to human uroepithelial cells . In addition, the ucaA gene encoding the structural component of UCA pili was subcloned, and its DNA sequence was determined . Amino acid sequence homology (30 to 50%) was found between mature UcaA protein and pilins from pathogenic bacteria representing several genera, including E . coli F17, G, and type 1C pilins, Haemophilus M43 pilin, and a Bordetella pilin. Infect Immun, 1995 May, 63(5), 2012 - 20 Inducible immunity with a pilus preparation booster vaccination in an animal model of Haemophilus ducreyi infection and disease; Desjardins M et al.; Using the temperature-dependent rabbit model of Haemophilus ducreyi infection as a quantitative virulence assay, we tested the abilities of two bacterial antigen preparations to induce protection against subsequent infection and disease . Lipooligosaccharide (LOS) and a pilus preparation were purified from H . ducreyi 35000 and were used in a booster immunization procedure . The serologic response to each immunogen was monitored by enzyme immunoassay . H . ducreyi virulence was assayed by intraepithelial inoculation and subsequent measurement of disease for homologous strain 35000 or clinical isolate RO-34 . LOS and the pilus preparation induced humoral responses . The kinetics of the LOS antibody response suggest a type 1 T-independent response, whereas the pilus preparation induced an anamnestic response . An inoculum of 10(5) CFU of H . ducreyi 35000 or RO-34 consistently produced ulcerative chancroidal lesions in naive rabbit controls . Immunization with LOS did not modify the virulence of H . ducreyi 35000 . Immunization with the strain 35000 pilus preparation significantly reduced the severity of disease and the duration of infection and disease compared with controls, with either homologous or heterologous strain infection . The histology of lesions from pilus preparation-vaccinated rabbits compared with that of lesions from controls revealed accelerated lymphoid cell recruitment, more prominent plasma cell infiltrate, and reduction in subsequent histiocytic infiltration . We conclude that both LOS and the pilus preparation are immunogenic and that the latter induces homologous and heterologous strain protection in this animal model of infection and disease. Infect Immun, 1995 May, 63(5), 1906 - 13 Human immunoglobulin M paraproteins cross-reactive with Neisseria meningitidis group B polysaccharide and fetal brain; Azmi FH et al.; Three hundred fifty-nine serum samples from patients with immunoglobulin M (IgM) or IgG monoclonal gammopathies were tested for binding to the capsular polysaccharide (PS) of Neisseria meningitidis group B (MenB PS, poly-alpha{2-->8}-N-acetylneuraminic acid) . Of 159 IgM paraproteins, 7 (4.4%) were positive, compared with 0 of 200 IgG paraproteins (P < 0.05) . Since MenB PS reactivity was limited to the IgM paraproteins, the 159 IgM paraproteins were tested by enzyme-linked immunosorbent assay (ELISA) for reactivity with seven other bacterial PSs . None reacted with meningococcal A or C, Haemophilus influenzae type b, or Streptococcus pneumoniae type 3, 6, 14, or 23 PS . The specificity of the MenB PS-reactive antibodies was confirmed by demonstration of binding to N . meningitidis group B cells but not to a capsular PS-deficient mutant and by specific inhibition of binding to solid-phase MenB PS by soluble MenB PS in an ELISA . Five of five antibodies tested protected infant rats from bacteremia caused by Escherichia coli K1, an organism with a PS capsule that also is composed of poly-alpha{2-->8}-N-acetylneuraminic acid . Each of the seven MenB PS-reactive paraproteins had autoantibody activity as defined by binding to homogenates of calf brain in a radioimmunoassay . For six of the seven antibodies, binding to calf brain was inhibited by the addition of soluble MenB PS . Thus, approximately 4% of human IgM paraproteins have autoantibody activity to poly-alpha{2-->8}-N-acetylneuraminic acid, an antigen expressed in fetal brain and cross-reactive with the MenB capsular PS . The reason for this skewing of the IgM paraprotein repertoire toward reactivity with poly-alpha{2-->8}-N-acetylneuraminic acid antigenic determinants is unknown. Infect Immun, 1995 May, 63(5), 1754 - 61 Use of tissue culture and animal models to identify virulence-associated traits of Haemophilus ducreyi; Alfa MJ et al.; To identify virulence-associated properties of Haemophilus ducreyi, 34 strains of this sexually transmitted pathogen were evaluated for in vitro phenotypic characteristics of potential relevance to chancroid pathogenesis and for their ability to produce lesions in the temperature-dependent animal model for chancroid . Of the 34 strains tested, all but three produced a cytopathic effect on human foreskin fibroblasts (HFF) and all but six strains formed large microcolonies on HFF monolayers . A subset of 12 selected strains underwent more extensive analyses and, when evaluated for both their cytadherence kinetics and growth in the presence of HFF monolayers, it was found that several of these strains had a very limited ability to attach to HFF cells . When the same 12 strains were tested in the temperature-dependent rabbit model, only the seven strains which were positive in all of these in vitro-based tests readily produced lesions . In contrast, the five strains that were noted to be deficient in one or more of the phenotypic characteristics scored in the in vitro systems did not produce lesions . This association between the traits measured in vitro and the ability to produce dermal lesions was significant (P = 0.0012) . These results suggest that in vitro behavior may be used to predict the virulence potential of H . ducreyi strains . Moreover, the phenotypic characteristics described in this study are appropriate focal points for efforts to determine the molecular basis of the virulence of this pathogen. Infect Immun, 1995 May, 63(5), 1631 - 6 Novel lipoprotein expressed by Neisseria meningitidis but not by Neisseria gonorrhoeae; Yang QL et al.; The ppk gene, which codes for the enzyme polyphosphate kinase in Neisseria meningitidis strain BNCV, is preceded by an open reading frame coding for a protein with a predicted size of 19.2 kDa with a typical lipoprotein signal sequence of 21 amino acids . The protein has significant homology to the N-terminal portion of an outer membrane protein from Haemophilus somnus (J . Won and R . W . Griffith, Infect . Immun . 61:2813-2821, 1993) . Sequencing of the same open reading frame from meningococcus strain M1080 predicted an almost identical protein . Antisera were raised against the lipoprotein, expressed in Escherichia coli as a fusion protein with glutathione S-transferase . The antisera reacted with meningococcal membrane fractions on a Western blot (immunoblot) but did not elicit complement-dependent bactericidal activity . Restriction enzyme digestion demonstrated conservation of this portion of the meningococcal and gonococcal chromosomes . However, antisera raised to the recombinant protein showed that the protein was absent from all strains of gonococcus tested . The sequences of the gene from several strains of Neisseria gonorrhoeae and N . meningitidis were compared and found to be almost identical, except that the coding sequences from all of the gonococcal strains were terminated prematurely as a result of a frameshift mutation . The significance of the remarkable conservation of these gonococcal genes is discussed. J Infect, 1995 May, 30(3), 219 - 22 Expression of capsules by Haemophilus influenzae in mixed infections; Brook I et al.; The pathogenicity of eight clinical isolates of non-type b Haemophilus influenzae was investigated by inoculating them subcutaneously into mice, alone or mixed with viable or non-viable bacteria of certain other species . Three of the H . influenzae isolates were non-capsulated while five were slightly capsulated (less than 1% of organisms had capsules) . The other strains of bacteria tested were four isolates of capsulated and four isolates of non-capsulated pigmented strains of Prevotella sp . and Porphyromonus sp . as well as a capsulate Klebsiella pneumoniae ("helpers") . None of the non-capsulated strains induced an abscess when inoculated alone . Following co-inoculation of viable or non-viable "helpers" with H . influenzae, abscesses were formed in all instances in which the "helper" had a capsule . Profusely capsulated cells of H . influenzae were recovered, however, only from abscesses induced with the five slightly capsulated strains of H . influenzae . These capsulated organisms were found serologically to be of type b and induced abscesses when inoculated alone . Our findings illustrate the ability of non-capsulated strains of H . influenzae to produce progeny of capsulated type b organisms after co-inoculation with certain other species. J Hosp Infect, 1995 May, 30(1), 31 - 7 The relationship between intraoperative contamination of the lower respiratory tract and postoperative chest infection; Morran GG et al.; The relationship between intraoperative contamination of the lower respiratory tract and postoperative chest infection was studied in 193 patients undergoing biliary tract surgery . During surgery, sputum was obtained from the lower respiratory tract for bacteriological culture . The diagnosis of postoperative pulmonary complications was based on clinical criteria, supported by the pattern of sequential blood gas changes in the postoperative period . Chest infection was present in 30% of patients who harboured Haemophilus species in their sputum at the time of surgery compared with 10% of those with negative cultures . Contamination of the lower respiratory tract at operation by Haemophilus sp . is associated with development of postoperative chest infection. Clin Diagn Lab Immunol, 1995 May, 2(3), 286 - 90 A rapid and sensitive chemiluminescence assay for evaluation of functional opsonic activity of Haemophilus influenzae type b-specific antibodies; Ojo-Amaize EA et al.; Luminol-enhanced chemiluminescence (CL) of heterologous neutrophils was used to assess the capacity of a 1-ng/ml concentration of Haemophilus influenzae type b (Hib)-specific antibodies to induce opsonization of Hib with autologous heat-inactivated sera from children immunized with Hib capsular polysaccharide-polyribosylribitolphosphate (Hib-PRP) conjugate vaccine . Serum samples from 15 of 36 children (42%) vaccinated with Hib-PRP conjugate vaccine had protective levels of Hib-specific antibodies of > or = 1,000 ng/ml . Ten of these 15 (67%) had poor or nonfunctional opsonic activity . Of the 10 children whose sera lacked opsonic activity, 5 (50%) presented with recurrent Hib infection . In contrast, none of the sera of 20 healthy adults lacked opsonic capability . CL intensity was proportional to the concentration of anti-Hib antibodies used for opsonization . Furthermore, the titers of Hib-PRP-specific antibody in children and adults did not correlate with opsonic activity . These results suggest that luminol-enhanced CL as described here with minute concentrations of antibody for opsonization can be used to assess functional capacity of anti-Hib antibodies after vaccination or natural infection in the evaluation of patients with recurrent infections. Clin Diagn Lab Immunol, 1995 May, 2(3), 272 - 6 Effect of age on concentrations of serum antibodies to viral, bacterial, and food antigens in elderly Swiss people; Brussow H et al.; Serum antibody concentrations to two viral, five bacterial, and two food antigens were investigated in 307 elderly Swiss subjects, and the hypothesis of whether serum antibody titers decreased with age was tested . The cross-sectional part of the study consisted of 216 unselected consecutive patients hospitalized in one geriatric hospital . The patients were divided into two age groups (65 to 84 and 85 to 102 years old), and their antibody titers were compared . No age-related decreases in antibody titers were observed . The members of the two age groups were well matched for medical diagnosis and nutritional and inflammatory status . The prospective part of the study consisted of 91 healthy elderly subjects living in the community; they were 71 to 76 years old when they were enrolled in the study . Their serum antibody status was measured at the beginning of the study and 4 years later . We observed a significant decrease in diphtheria antitoxin levels and a significant increase in antibody titer to the capsular polysaccharide of Streptococcus pneumoniae . No change in antibody titer to rotavirus, respiratory syncytial virus, lipopolysaccharide of Escherichia coli, C polysaccharide of S . pneumoniae, or the polyribosyl-ribitol phosphate of Haemophilus influenzae was observed . Thus, no signs of B-cell immunosenescence were seen in these two groups of elderly Swiss people. Eur Respir J, 1995 May, 8(5), 709 - 14 Interaction of fimbriated and nonfimbriated strains of unencapsulated Haemophilus influenzae with human respiratory tract mucus in vitro; Barsum W et al.; Adherence to mucus may influence bacterial colonization of the respiratory tract . Clinical isolates of nontypable Haemophilus influenzae (NTHi) from the respiratory tract are often fimbriated . We wondered whether fimbriated strains have a different adherence from related nonfimbriated strains . A microtitre plate assay has been developed to study adherence of nontypable H . influenzae to mucus . Wells were coated by incubation either with sol phase of sterile mucoid secretions or with purified preparations of mucins . Two laboratory pairs of fimbriated (F+) and nonfimbriated (F-) nontypable H . influenzae, and six fresh clinical isolates of fimbriated nontypable H . influenzae each with nonfimbriated partners derived by serial passage on agar, were cultured to mid-log phase, washed, and then added to the wells . They were then incubated at 37 degrees C for 30 min before washing to remove unbound bacteria . Adherent bacteria were desorbed by agitation with 0.5% Tween 80 and a viable count performed . The two fimbriated laboratory strains (n = 12 and n = 17), and 5 of the 6 fimbriated clinical isolates were more adherent to sol phase than their respective nonfimbriated partners . Two nonfimbriated clinical isolates were more adherent to plastic than their fimbriated partners . A fimbriated laboratory strain was more adherent than its nonfimbriated partner both to a purified preparation of high molecular mass mucin and to the glycopeptide fraction of the same . We conclude that fimbriated strains of nontypable H . influenzae have increased adherence to sol phase of mucus and purified human respiratory tract mucin . The interactions of fimbriae with mucus are likely to be complex, and may involve both nonspecific and specific interactions. Pediatr Infect Dis J, 1995 May, 14(5), 445 - 9 Present and future challenges of immunizations on the health of our patients; Gershon AA; A recent analysis demonstrated a change in incidence approaching 100% for diseases against which we routinely immunize in the United States . At present, measles, mumps, rubella, invasive Haemophilus disease, poliomyelitis, diphtheria and tetanus are well-controlled but not eliminated . Diseases that now pose special problems include pertussis, hepatitis A and B and varicella . The incidence of pertussis surged in 1994, possibly in part because of waning immunity in the immunized population . Acellular pertussis vaccines are available for booster doses in children but are not now recommended for adults . Licensure of acellular pertussis vaccines for primary immunization of infants is eagerly awaited . Recombinant hepatitis B vaccine has been licensed for more than 10 years but there has been little change in disease incidence in the United States . Routine immunization of infants is now recommended but concerns exist about cost and persistence of immunity into adolescence . Inactivated hepatitis A vaccines appear to be highly effective in preventing clinical hepatitis and controlling epidemics . Potential target populations include military personnel, day-care attendees and travelers . Hepatitis A vaccine may be recommended for all children after approval by the United States Food and Drug Administration and if a combination vaccine becomes available . A live, attenuated varicella vaccine developed in 1974 and unlicensed in the United States is safe and highly effective in preventing varicella in healthy and immunocompromised populations . It also appears to reduce subsequent development of herpes zoster . Vaccines against pneumococci (conjugate vaccine), respiratory syncytial virus, rotavirus, tuberculosis and human immunodeficiency virus are needed . Research and technology to develop these vaccines must be developed, and efficient delivery mechanisms must be created and implemented. Pediatr Infect Dis J, 1995 May, 14(5), 420 - 3 Resistance among problem respiratory pathogens in pediatrics; Doern GV; During the past two decades, the prevalence of beta-lactamase production with nontypable strains of Haemophilus influenzae has increased to about 35% . Fortunately, rates of resistance to other oral antimicrobials have not developed at a comparable pace . Amoxicillin/clavulanate, cefuroxime and cefpodoxime remain nearly uniformly active whereas rates of resistance to tetracycline, trimethoprim/sulfamethoxazole, chloramphenicol, cefaclor, loracarbef, cefprozil, azithromycin and clarithromycin remain low (1 to 5%) . Virtually all clinical isolates of Moraxella catarrhalis produce beta-lactamase and are probably resistant to ampicillin and amoxicillin . However, alternative oral antimicrobials are almost always active . A compelling problem facing pediatricians today is the emergence of penicillin resistance with clinical isolates of Streptococcus pneumoniae . Currently, 15 to 25% of pneumococcal isolates in the United States have either intermediate (10 to 20%) or complete (3 to 5%) penicillin resistance caused by alterations in penicillin-binding proteins . Loss of activity of other beta-lactams is observed with penicillin-resistant S . pneumoniae . Third generation cephalosporins retain sufficient activity to warrant use in selected pneumococcal infections, even those caused by completely penicillin-resistant strains . Unfortunately, strains of S . pneumoniae with further alterations in penicillin-binding proteins have emerged such that even extended spectrum third generation cephalosporins lack activity . Rates of resistance to non-beta-lactam agents are also changing . The consequence of these changing patterns of resistance is that therapeutic options for pneumococcal infections in some patients are becoming increasingly limited. Pediatr Infect Dis J, 1995 May, 14(5), 415 - 9 Antimicrobial therapy issues facing pediatricians; Klein JO; In the field of infectious diseases, the emergence of new pathogens or old diseases in newly recognized forms; changing virulence of pathogens; changing patterns of antimicrobial susceptibility; new diagnostic techniques, drugs or vaccines; changing concepts of chemoprophylaxis; controversies about medical vs . surgical techniques; and the challenge of care of children with infectious diseases within new guidelines of managed care are recently identified areas of change . The increased resistance of Streptococcus pneumoniae to many commonly used antimicrobials and the increased proportion of beta-lactamase-producing nontypable Haemophilus influenzae and Moraxella catarrhalis concern many practitioners . The decreased antibiotic susceptibility of S . pneumoniae is a relatively new phenomenon in the United States . Optimal therapy for mild, moderate or severe pneumococcal disease is dependent on current local susceptibility patterns . Group A streptococci are uniformly susceptible to readily achieved concentrations of all penicillins and cephalosporins . However, recent clusters of cases of rheumatic fever, increased recognition of toxic shock syndrome and bacteremic and localized severe pneumococcal disease have increased concern about the changing ecology of the Streptococcus and the implications for therapy . Finally recognition that many children with acute bacterial otitis media have resolution of disease without use of antimicrobial agents has led to more rigorous study designs for evaluating new drugs.(ABSTRACT TRUNCATED AT 250 WORDS) Pediatr Infect Dis J, 1995 May, 14(5), 350 - 4 The immunogenicity of Haemophilus influenzae type b conjugate (HbOC) vaccine in human immunodeficiency virus-infected and uninfected infants; Kale KL et al.; Enzyme-linked immunosorbent assay polyribosyl ribitol phosphate (PRP) antibody responses to Haemophilus influenzae type b conjugate vaccine (HbOC) given at 2, 4 and 6 months of age were retrospectively compared in 23 human immunodeficiency virus (HIV) and 24 non-HIV-infected infants . HIV-infected infants were divided into those who were P1 (asymptomatic) or P2 (symptomatic) by 1 year of age . The P2 group was further divided into P2A (mildly symptomatic) and > P2A (rapidly symptomatic) by 1 year of age . The post-third HbOC dose geometric mean antibody titer to PRP was significantly lower in 12 P2 infants (0.43 microgram/ml) than either the 11 P1 infants (5.03 micrograms/ml, P < 0.05) or the 24 non-HIV infected infants (3.43 micrograms/ml, P < 0.05) . Within the P2 group, the geometric mean antibody titer to PRP was significantly higher in 5 P2A infants (1.63 micrograms/ml) compared with 7 infants who were > P2A (0.17 microgram/ml, P < 0.05) . After the third HbOC dose, PRP antibody titers were > or = 1.0 micrograms/ml for 4 of 12 P2 compared with 9 of 11 P1 infants (P < 0.05) . Within the P2 group, PRP antibody titers were > 1.0 micrograms/ml for 4 of 5 P2A compared to 0 of 7 infants who were > P2A (P < 0.05) . HIV-infected infants with PRP antibody titers > or = 1.0 micrograms/ml after the third HbOC dose had significantly higher mean CD4 counts (2842 cells/mm3) at the time of the third HbOC dose than those with lower PRP titers (1655 cells/mm3) (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) Jpn J Antibiot, 1995 May, 48(5), 602 - 9 {Antibacterial activities of cefmenoxime against recent fresh clinical isolates from patients in sinusitis}; Yokota N et al.; In order to evaluate antimicrobial activity of cefmenoxime (CMX), minimum inhibitory concentrations (MICs) of CMX and control drugs were determined against clinical isolates from patients of sinusitis that were obtained in our laboratory from October of 1993 to March of 1994 . The results are summarized as follows; 1 . CMX showed strong antimicrobial activities against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella subgenus Branhamella catarrhalis that were 3 major aerobic bacteria from sinusitis . Antimicrobial activities of CMX against benzylpenicillin (PCG)-insensitive S . pneumoniae (PISP) and PCG-resistant S . pneumoniae (PRSP) were stronger than those of ampicillin (ABPC), and these strong activities suggested that CMX might have strong antimicrobial activities against beta-lactamase producing H . influenzae and M . (B.) catarrhalis . 2 . Antimicrobial activities of CMX against microaerophiles, Streptococcus constellatus, Streptococcus intermedius and Gemella morbillorum and against Peptostreptococcus spp., from chronic sinusitis and odontogenic maxillary sinusitis, were stronger than those of most of the control drugs . 3 . The MIC90's of CMX against isolates from patients of sinusitis were < or = 0.025-0.39 micrograms/ml . These values were lower than transitional concentrations in mucous membrane of maxillary sinus obtained when "1% CMX nasal solution" was used with nebulizer . It appears likely that sufficient concentrations exceeding MICs against main organisms would be obtained by nebulizer treatment using CMX nasal solution. Clin Infect Dis, 1995 May, 20(5), 1381 - 3 Haemophilus paraphrophilus endocarditis: case report and review; Coll-Vinent B et al.; Endocarditis due to Haemophilus paraphrophilus is an uncommon disease . We report a case of H . paraphrophilus endocarditis with embolic complications in which the causative organism was resistant to beta-lactam antibiotics . Before April 1994, 16 cases of H . paraphrophilus endocarditis had been reported . Infection by this organism usually affects a previously damaged mitral valve . We emphasize the fastidiousness of the organism and the high incidence of embolic complications, which determine the outcome . To our knowledge we describe the first patient with endocarditis due to beta-lactam-resistant H . paraphrophilus. Clin Infect Dis, 1995 May, 20(5), 1164 - 8 A single daily dose of ceftriaxone for bacterial meningitis in adults: experience with 84 patients and review of the literature; Cabellos C et al.; Although the pharmacokinetics of ceftriaxone allows its administration in a single daily dose, this practice is not standard in the treatment of bacterial meningitis . Herein, we review our experience and that of other investigators with this mode of therapy . We used a single daily dose of ceftriaxone (50 mg/{kg.d}; maximum, 4 g/d) for the treatment of bacterial meningitis in 84 adult patients . Meningitis was due to Neisseria meningitidis in 34 cases, to Streptococcus pneumoniae in 25, to Escherichia coli in three, to Klebsiella pneumoniae in two, to Haemophilus influenzae in two, to viridans streptococci in two, and to an unknown agent in 16 . Eleven patients died, for an overall mortality of 13%; therapy failed in three additional cases . The mean trough levels of ceftriaxone in cerebrospinal fluid was 3.5 micrograms/mL; the median trough bactericidal titer at this site was 1:128 . Both our experience and that in the literature suggest that a single daily dose is optimal when ceftriaxone is used for the treatment of bacterial meningitis. Arch Dis Child, 1995 May, 72(5), 432 - 4 Urinary excretion of cortisol after immunisation; Westaway J et al.; Urinary cortisol excretion and rectal temperature were measured in 66 infants before and after immunisation against diphtheria, tetanus, pertussis, and Haemophilus influenzae type b . Immunisation produced a significant increase of rectal temperature the next night at all ages . Infants without an adult-like night time body temperature pattern had a significant increase in urinary cortisol excretion night and morning after immunisation . Once an adult-like night time body temperature pattern developed immunisation no longer significantly raised urinary cortisol output. Zhonghua Min Guo Xiao Er Ke Yi Xue Hui Za Zhi, 1995 May-Jun, 36(3), 164 - 9 IgG subclasses in childhood infections; Bradwell AR; Selective IgG subclass (IgGSc) deficiencies are frequently found in association with recurrent infections in childhood . IgG1 deficiency is the most severe and is associated with features typical of panhypogammaglobulinaemia . Immunoglobulin replacement therapy is usually required . IgG2 deficiency is associated with recurrent infections with encapsulated bacteria such as Haemophilus influenzae and Streptococcus pneumoniae . IgG2 deficiency may be transient in children under five years of age and patients improve with antibiotics and immunisation . IgG3 and IgG4 deficiency are commonly found in children with recurrent infections and may indicate a disordered immune system since absence of these antibodies alone appears insufficient to cause symptoms . Children may also have selective IgGSc deficiencies in the absence of recurrent infections . This is explained by compensatory factors in other parts of the immune system . Measurement of IgGSc levels should be based on highly specific polyclonal antisera which show no IgGSc cross-reactivity . Most monoclonal antibodies are unsatisfactory since allotypes are detected variably, leading to excess reporting of IgGSc deficiencies and Mabs cannot be used for nephelometric or turbidimetric methods. J Clin Microbiol, 1995 May, 33(5), 1426 - 7 Evaluation of novel vancomycin-containing medium for primary isolation of Kingella kingae from upper respiratory tract specimens; Yagupsky P et al.; A new selective medium (BAV), consisting of trypticase agar with 5% sheep hemoglobin and 2 micrograms of vancomycin per ml, was compared with the routine blood-agar medium for the primary isolation of Kingella kingae from upper respiratory specimens from a population of young children . Infection was detected by the BAV medium in 43 of 44 (98%) cultures positive for K . kingae, and detection of the organism was facilitated by inhibition of gram-positive flora . Infection was detected in only 10 of 44 (23%) positive cultures by the blood-agar medium, and plates were usually covered by abundant normal flora, making the recognition of K . kingae much more difficult . Challenge of the medium with different organisms of respiratory origin showed that the BAV medium was inhibitory for gram-positive cocci and Haemophilus influenzae but that it supported growth of eight K . kingae strains isolated from patients with invasive infections . The new medium appears to be a useful epidemiological tool for studying the respiratory carriage of K . kingae. J Clin Microbiol, 1995 May, 33(5), 1174 - 9 PCR amplicon restriction endonuclease analysis of the chromosomal dhps gene of Neisseria meningitidis: a method for studying spread of the disease-causing strain in contacts of patients with meningococcal disease; Kristiansen BE et al.; We tested two sets of primers derived from the dhps gene of Neisseria meningitidis for the amplification of meningococcal DNA by PCR . Both the NM1-NM6 primers and the NM3-NM6 primers amplified dhps DNA from all of the meningococci included in the study, resulting, in most cases, in amplicons of 0.70 and 0.23 kb, respectively . Also, dhps DNAs of N . gonorrhoeae and some commensals were amplified but Haemophilus influenzae, Streptococcus pneumoniae, and Escherichia coli DNAs were not . By PCR amplicon restriction endonuclease analysis (AREA) of the larger amplicon, we could differentiate between individual strains of N . meningitidis . Following two cases of meningococcal disease, we used PCR AREA to identify healthy contacts carrying the disease-causing strain . We conclude that PCR AREA is a useful method for meningococcal strain differentiation and that it has potential as a method for studying the spread of a disease-causing strain in an affected population . The method is quicker and easier to perform and interpret than chromosomal DNA fingerprinting. Thorax, 1995 May, 50(5), 543 - 7 Aetiology of community-acquired pneumonia: a prospective study among adults requiring admission to hospital; Bohte R et al.; BACKGROUND--The prevalence of microorganisms causing community-acquired pneumonia in patients who required admission to hospital was investigated and the percentage of cases whose aetiology remained unknown due to the study design and logistical problems estimated . METHODS--Between January 1991 and April 1993 all patients with community-acquired pneumonia admitted to six hospitals were included in the study . Aetiological diagnosis, categorised as definite, probable and possible, was based on the results of routine microbiological and serological tests . RESULTS--Three hundred and thirty four patients with a median age of 65 (range 17-92) years were enrolled in the study . The diagnosis of community-acquired pneumonia was definite in 108 cases, and probable or possible in 73 and 27 cases, respectively, including dual infections . Streptococcus pneumoniae was the predominant pathogen (27%) followed by viruses and Haemophilus influenzae (both about 8%) and Mycoplasma pneumoniae (6%) . Chlamydia spp (3%) and Legionella pneumophila (2%) were less frequently detected . No diagnosis was made in 45% of the cases . With adjustment for anti-microbial therapy before admission and for other logistical considerations, it is estimated that the aetiology could have been ascertained in 65% of the cases . CONCLUSIONS--Streptococcus pneumoniae is the most frequently detected cause of community-acquired pneumonia . The inability to detect a micro-organism results mainly from the use of routine diagnostic tests and, to a lesser extent, from logistical problems or the use of antibiotics before admission. J Antimicrob Chemother, 1995 May, 35(5), 681 - 6 The in-vitro activity of biapanem against 964 clinical isolates of aerobic bacteria; Raymond NJ et al.; The in-vitro susceptibilities of 964 clinical isolates of aerobic bacteria to biapenem (a novel carbapenem), imipenem, ceftazidime and ciprofloxacin were determined by an agar dilution method . Compared with imipenem, biapenem exhibited greater activity against aerobic Gram-negative bacilli, similar activity against Neisseria meningitidis, Neisseria gonorrhoeae and Haemophilus influenzae and less activity against Gram-positive cocci. Diagn Microbiol Infect Dis, 1995 May-Jun, 22(1-2), 147 - 54 Review and reassessment of dosing schedules for cefotaxime in selected medical indications; Young LS; Cefotaxime, the first widely used "third-generation" cephalosporin, has established efficacy against a variety of serious bacterial pathogens . Some of the initial clinical studies in the United States using this agent employed large doses of the compound, up to 12 g/day, for adults . In contrast, however, initial European studies were largely with low doses of 1 to 2 g every 12 h . In the recent past, however, an effort has been made, both in the United States and in Europe to reevaluate the dosage of cefotaxime . In various clinical studies, lower doses of cefotaxime have been successfully employed for infections of the urinary tract, peritoneum, biliary tract, lung, and skin and soft tissues . The results of a number of these studies will be reviewed, including a large postmarketing surveillance study carried out in Germany during 1992 . The results suggest that cefotaxime doses as low as 1 g, at intervals as long as every 12 h, can be adequate for treatment of the most commonly encountered infections, such as those caused by some hemolytic streptococci, Staphylococcus aureus, Haemophilus spp., and enteric bacilli in nonimmunocompromised patients. Diagn Microbiol Infect Dis, 1995 May-Jun, 22(1-2), 125 - 7 Update on the use of cefotaxime for pediatric meningitis in Portugal; Lecour H et al.; We treated 256 children who had identified bacterial meningitis with cefotaxime . Causative organisms were: Neisseria meningitidis in 108 cases, Streptococcus pneumoniae in 61, Haemophilus influenzae in 60, enteric Gram-negative bacilli in 21, and Staphylococcus spp . in six . Daily doses of cefotaxime were 150-200 mg/kg . A total of 240 patients (93.7%) were cured . In the cured patients, sterilization of cerebrospinal fluid was obtained in the first 72 h of treatment in 214 (80.0%) . Cefotaxime is an effective and safe drug for the treatment of childhood bacterial meningitis. Diagn Microbiol Infect Dis, 1995 May-Jun, 22(1-2), 105 - 10 Cefotaxime use in pediatric infections; Dajani AS; Cefotaxime has been used extensively in many pediatric centers in the United States for the past 10 or more years . Its main usage has been for the treatment of various serious bacterial infections in pediatric patients, primarily meningitis and sepsis . It has also been used to treat intraabdominal, urinary tract, soft tissue, bone, and joint infections . Although there has been a marked reduction in the incidence of invasive Haemophilus influenzae type b infections following the introduction of effective vaccines, cefotaxime remains very useful against the other common pathogens causing serious infections in pediatric patients . The increasing number of pneumococci resistant to penicillin and third-generation cephalosporins has created a new challenge for the management of serious pneumococcal infections . In many institutions, cephalosporins in general have been overused and abused, resulting in the emergence of resistant organisms and an increasing burden on resources . The judicious use of cefotaxime and other cephalosporins should be emphasized. J Clin Microbiol, 1995 May, 33(5), 1192 - 5 Long PCR-ribotyping of nontypeable Haemophilus influenzae; Smith-Vaughan HC et al.; PCR-ribotyping, a new typing method based on long PCR, has been developed for nontypeable Haemophilus influenzae (NTHi) . Ribosomal operons of NTHi were amplified by long PCR and were found to be highly polymorphic for internal HaeIII sites . The technique was applied to 49 isolates previously subjected to conventional ribotyping, and the two methods showed a high level of concordance for serial isolates from individual subjects . PCR-ribotyping provides a powerful new typing tool for strain characterization in epidemiological investigations of NTHi. Ned Tijdschr Geneeskd, 1995 Apr 29, 139(17), 885 - 90 {Pediatric surveillance of invasive infections caused by Haemophilus influenzae type b in children in the period following introduction of vaccination}; Conyn-van Spaendonck MA et al.; OBJECTIVE . Evaluation of the effect of vaccination against Haemophilus influenzae type b (Hib) on the occurrence of invasive Hib infections in children since its introduction into the national immunization programme in April 1993 . DESIGN . Observational study . SETTING . Nationwide investigation . METHOD . Data collected through active surveillance of invasive Hib infections by paediatricians for the period from October 1993 to September 1994 (11 months) were compared with data from the meningitis surveillance by the Netherlands Reference Laboratory for Bacterial Meningitis . RESULTS . A total of 139 paediatric reports of invasive disease by H . influenzae concerned 57 cases of only meningitis, 35 of meningitis with sepsis, 2 of meningitis with arthritis, one of meningitis with arthritis and osteomyelitis, 34 of epiglottitis including one case with sepsis, 8 of only sepsis and 2 of only arthritis . All proven infections by Hib occurred in children who had not or incompletely been vaccinated . One child with sepsis had had three vaccinations and became ill five months later; the isolated bacterial strain was not serotyped . Typing was performed in only 80% of the isolates, of which 98% were of type b . Appropriate culturing and typing was often omitted in case of epiglottitis . CONCLUSION . The effect of vaccination against Hib became apparent in a small number of cases of invasive Hib disease reported by paediatricians; the peak incidence of meningitis no longer occurred in children under one year of age but in children aged one year . The paediatric surveillance described offers possibilities for monitoring Hib epidemiology. Ned Tijdschr Geneeskd, 1995 Apr 29, 139(17), 880 - 4 {Absence of meningitis caused by Haemophilus influenzae type b in The Netherlands following twofold vaccination}; van Alphen L et al.; OBJECTIVE . To determine the two-year results of nationwide vaccination with Haemophilus influenzae type b (Hib) vaccine on the occurrence of Hib meningitis in the Netherlands . DESIGN . Retrospective controlled study . SETTING . The Netherlands . METHOD . Children born since April 1, 1993 are vaccinated at the age of 3, 4, 5 and 11 months to protect them from infections with Hib . The number of Hib meningitis patients in the period 1 April, 1993 to 1 April, 1995, among infants born in this period who were offered the Hib vaccine (study group), was compared with the number of Hib meningitis patients in the period 1 April, 1991 to 1 April, 1993 among children born in last-mentioned period (control group) . RESULTS . Twenty-one cases of meningitis by Hib were observed in the study group . Twelve children who, as a consequence of their age, had only been vaccinated once or not at all; 7 children were not vaccinated for several reasons . In addition one patient was infected by H . influenzae type f strain and one by a non-typable strain . In the control group 185 cases of Hib meningitis occurred . CONCLUSION . Hib meningitis was not observed among infants who had been vaccinated at least twice. J Immunol, 1995 Apr 15, 154(8), 4195 - 202 Functional differences in idiotypically defined IgG1 anti-polysaccharide antibodies elicited by vaccination with Haemophilus influenzae type B polysaccharide-protein conjugates; Lucas AH et al.; We investigated the relationship between the form of the Haemophilus influenzae type B (Hib) polysaccharide (PS)-protein conjugate vaccine, Id expression, and Ab quality . Two post-vaccination pools were prepared from sera of infants vaccinated with either Hib PS oligomers coupled to CRM197, a mutant diphtheria toxin (HbOC), or with higher m.w . Hib PS coupled to an outer membrane protein complex of Neisseria meningitidis group B (Hib-OMP) . The mean anti-Hib PS Ab avidity of the serum pool from the infants vaccinated with HbOC was threefold higher than that of the pool from infants vaccinated with Hib-OMP . Using sequential immunoabsorption, three IgG1 idiotypically-defined anti-Hib PS fractions were isolated from each of the serum pools: Hibld-1, Hibld-2, and a Hibld-1/-2-depleted population, designated Hibld-0 . Hibld-1 and Hibld-2 are idiotypic markers for anti-Hib PS Abs expressing kappa II-A2 and lambda VII V regions, respectively . Hibld-1 anti-Hib PS Abs had significantly higher avidity, 2- to 19-fold higher in vitro bactericidal activity, and were more protective against Hib bacteremia in infant rats, than the respective Hibld-2 Abs isolated from each of the pools . Comparing the two vaccines, Hibld-1 anti-Hib PS Abs elicited by HbOC had significantly higher avidity and 10-fold higher bactericidal and rat protective activity than the Hibld-1 Abs elicited by Hib-OMP . These findings demonstrate that the molecular form of the Hib PS immunogen dictates both V region usage and quality of Ab function. Gene, 1995 Apr 14, 156(1), 97 - 9 The sequencing of the 80-kDa D15 protective surface antigen of Haemophilus influenzae; Flack FS et al.; The 80-kDa D15 antigen (D-15-Ag) has previously been shown to be a target for protective immunity and conserved amongst typeable and nontypeable Haemophilus influenzae . Here, the gene encoding D-15-Ag is shown to encode a 797-aa polypeptide which, after cleavage of the predicted signal peptide, would have a molecular mass of 85,632 Da. Proc Natl Acad Sci U S A, 1995 Apr 11, 92(8), 3616 - 20 Identification of a DNA transformation gene required for com101A+ expression and supertransformer phenotype in Haemophilus influenzae; Zulty JJ et al.; DNA sequencing, RNA mapping, and protein expression experiments revealed the presence of a gene, tfoX+, encoding a 24.9-kDa polypeptide, that is transcribed divergently from a common promoter region with the Haemophilus influenzae rec-1+ gene . H . influenzae strains mutant for tfoX failed to bind transforming DNA and were transformation deficient . Primer extension experiments utilizing in vivo total RNA from precompetent and competent H . influenzae cells demonstrated that transcription of tfoX+ increased immediately upon competence induction, suggesting that tfoX+ is an early competence gene . Similar experiments showed that the expression of the late competence-specific gene, com101A+, was tfoX+ dependent . Moreover, expression of plasmid-borne tfoX+ in H . influenzae resulted in constitutive competence . The addition of cyclic adenosine monophosphate (cAMP) to strains carrying a tfoX::lacZ operon fusion resulted in an immediate increase in beta-galactosidase activity that correlated with an increase in genetic transformability . Collectively, our results suggest that TfoX may play a key role in the development of genetic competence by regulating the expression of late competence-specific genes. Scand J Immunol, 1995 Apr, 41(4), 324 - 30 Predominant V-region gene configurations in the human antibody response to Haemophilus influenzae capsule polysaccharide; Pinchuk GV et al.; The antibody response to Haemophilus influenzae type b polysaccharide (Hib PS) is known to be encoded by a few V-region genes . We have obtained four human monoclonal Hib PS antibodies from four healthy adult subjects immunized with diphtheria toxin-conjugated Hib PS vaccine . The VH gene segments that encode for these antibodies belong to the VH3 gene family, of which two are related to the V3-23 gene and two to the VH3b subfamily . Both hybridomas that express a V3-23-related gene use short D-segments (3 bp), the JH6 gene segment and a V kappa gene derived from the A2 germline gene . The two hybridomas that express VH3b genes use D-segments of conventional length (24-33 bp), the JH4 gene segment and a non-A2 V kappa gene . Comparison of our sequences with those reported by others suggests that the above patterns of V-region gene segment association exemplify two V-region gene configurations that are predominant in the Hib PS antibody response . The first configuration is reminiscent of antibodies produced by B-1 B cells while the second is more characteristic of antibodies produced by conventional B cells . The possibility that these two configurations, in fact, represent the products of two different B cell lineages remains to be elucidated. J Bacteriol, 1995 Apr, 177(7), 1788 - 96 Genomic organization of the Klebsiella pneumoniae cps region responsible for serotype K2 capsular polysaccharide synthesis in the virulent strain Chedid; Arakawa Y et al.; The genomic organization of the chromosomal cps region that is responsible for capsular polysaccharide synthesis in Klebsiella pneumoniae Chedid (O1:K2) was investigated . Deletion analyses and Southern hybridization studies suggested that the central region of the cloned 29-kb BamHI fragment is indispensable for K2 capsular polysaccharide synthesis . The 24,329-bp nucleotide sequence of the Klebsiella cps region was determined and deposited in the EMBL and GenBank databases through DDBJ and assigned accession number D21242 . Nineteen possible open reading frames (ORFs) were identified in the sequenced area . Among them, 13 ORFs are very close to each other . Six of the 19 ORFs show considerable nucleotide sequence similarities to Salmonella typhimurium cpsG, cpsB, rfbP, and orf2.8, Escherichia coli gnd, and Haemophilus influenzae bexD, respectively . Moreover, the deduced amino acid sequence of the ORF10 product demonstrated a highly hydrophobic profile and showed putative membrane topology similarity to Rickettsia prowazekii ATP/ADP translocase . Nucleotide sequence similar to the sigma 54-dependent promoter, as well as the usual -35 and -10 sequences, were identified just upstream of ORF3, which is the first ORF in the polycistronic structure . Furthermore, a sequence (GGGCGGTAGCGT) found just downstream of the sigma 54-dependent promoter-like sequence was generally conserved among gene clusters implicated in cell surface polysaccharide synthesis, such as Salmonella rfb and viaB and E . coli kpsMT and rfaQPG . A possible transcriptional terminator with a hairpin loop structure found just downstream of ORF15 that is a homolog of E . coli gnd . K2 capsular polsaccharide biosynthesis in E . coli K-12 depends on cpsB (mannose-1-phosphate guanyltransferase gene), and Klebsiella cpsB, found in the downstream region of the polycistronic structure, was able to complement cpsB of E . coli . Results of transposon insertion and promoter-cloning analyses were consistent with the results of nucleotide sequence analysis. Infect Immun, 1995 Apr, 63(4), 1329 - 35 Complement-independent binding of microorganisms to primate erythrocytes in vitro by cross-linked monoclonal antibodies via complement receptor 1; Powers JH et al.; Under certain circumstances, soluble antigens, particulate antigens, and/or microorganisms have been shown to bind to primate erythrocytes via complement receptor 1 (CR1) in the presence of specific antibodies and complement . This immune adherence reaction, specific for CR1, can lead to neutralization of antigens in the circulation and their subsequent clearance from the blood . The present experiments utilized cross-linked monoclonal antibody complexes (heteropolymers) with specificity for both CR1 and either 35S-labeled herpes simplex virus capsid or Haemophilus influenzae as prototype viral and bacterial particulate antigens, respectively . In each case, the respective specific heteropolymers facilitated binding of the target antigens (> or = 70 to 90%) in vitro to erythrocytes in the absence of complement . Several experimental protocols were employed to demonstrate that heteropolymers mediate specific, rapid (> or = 30 s), and quantitative binding of prototypical particulate pathogens to human and monkey erythrocytes but not to sheep erythrocytes, which lack CR1 . These results extend the potential use of the erythrocyte-heteropolymer system to the neutralization and clearance of particulate viral and bacterial pathogens from the blood. Infect Immun, 1995 Apr, 63(4), 1241 - 5 Identification and purification of a conserved heme-regulated hemoglobin-binding outer membrane protein from Haemophilus ducreyi; Elkins C; A hemoglobin-binding protein (HgbA) from Haemophilus ducreyi was identified and purified . The 100-kDa HgbA was detected in all strains of H . ducreyi tested, and a somewhat larger hemoglobin-binding protein was found in one strain of Haemophilus influenzae . HgbA was purified and the amino acid sequence of the N terminus of HgbA revealed no significant homologies with known proteins . Two different antisera to HgbA from H . ducreyi 35000 recognized HgbA proteins from all tested H . ducreyi strains; they did not recognize proteins from the H . influenzae strain . Expression of HgbA was regulated by the level of heme but not by iron present in the medium . Animal species of hemoglobin competed with iodinated human hemoglobin for binding to whole cells of H . ducreyi and supported the growth of H . ducreyi . The lack of immunological cross-reactivity and the differences in hemoglobin specificities between the H . ducreyi and the H . influenzae hemoglobin-binding proteins suggest that they are unrelated. Infect Immun, 1995 Apr, 63(4), 1201 - 10 Identification and characterization of genes encoding the human transferrin-binding proteins from Haemophilus influenzae; Gray-Owen SD et al.; Haemophilus influenzae, a strict human pathogen, acquires iron in vivo through the direct binding and removal of iron from human transferrin by an as yet uncharacterized process at the bacterial cell surface . In this study, the tbpA and tbpB genes of H . influenzae, encoding the transferrin-binding proteins Tbp1 and Tbp2, respectively, were cloned and sequenced . Alignments of the H . influenzae Tbp1 and Tbp2 protein sequences with those of related proteins from heterologous species were analyzed . On the basis of similarities between these and previously characterized proteins, Tbp1 appears to be a member of the TonB-dependent family of outer membrane proteins while Tbp2 is lipid modified by signal peptidase II . Isogenic mutants deficient in expression of Tbp1 or Tbp2 or both proteins were prepared by insertion of the Tn903 kanamycin resistance cassette into cloned sequences and reintroduction of the interrupted sequences into the wild-type chromosome . Binding assays with the mutants showed that a significant reduction in transferrin-binding ability resulted from the loss of either of the Tbps and a complete loss of binding was evident when neither protein was expressed . Loss of either Tbp2 or both proteins correlated with an inability to grow on media supplemented with transferrin-bound iron as the sole source of iron, whereas the Tbp1+ Tbp2- mutant was able to grow only at high transferrin concentrations. Clin Infect Dis, 1995 Apr, 20 Suppl 1, S39 - 46 Recommendations for treatment of chancroid, 1993; Schulte JM et al.; Since the 1989 Sexually Transmitted Diseases Treatment Guidelines were published by the Centers for Disease Control and Prevention, changes in the efficacy of the recommended and alternative regimens for the treatment of Haemophilus ducreyi infections have been described . Among recommended agents, erythromycin remains effective, and although a single dose of ceftriaxone appears to remain effective in the United States, limited data from Kenya have shown that this regimen has been associated with treatment failures . Of alternative treatment regimens, trimethoprim-sulfamethoxazole has been associated with widespread failure, but little work has been done to further evaluate the efficacy of the amoxicillin/clavulanic acid and ciprofloxacin regimens . Of the new antimicrobials, azithromycin has been very effective in the United States, but the efficacy of this drug elsewhere has not been thoroughly evaluated . Fleroxacin has been very effective in Kenya . Data from Africa indicate that patients who are infected with the human immunodeficiency virus do not respond to therapy as well as patients who are not, and patients who are uncircumcised may not respond as well to therapy as do patients who are circumcised. Clin Infect Dis, 1995 Apr, 20 Suppl 1, S3 - 22 Early intervention for persons infected with human immunodeficiency virus; Branson BM; Early intervention for persons infected with human immunodeficiency virus (HIV) involves characterization of the stage of HIV disease, institution of therapy to prevent associated infections and postpone deterioration of immune function, and assistance in preventing transmission of the virus . This review examines the available data on the efficacy of current recommendations regarding the evaluation and management of persons with early HIV infection . Existing evidence supports the efficacy of physical examination, monitoring of the CD4+ cell count, tuberculin testing (with chemotherapy for persons who test positive), anergy testing, Papanicolaou testing and screening for gonorrhea and chlamydial infection (for high-risk women), screening for syphilis, antiretroviral therapy (for symptomatic patients), and guidance in reducing the transmission of HIV . Recommended measures for which evidence of clinical efficacy is less certain include immunization against infections due to influenza virus, Streptococcus pneumoniae, Haemophilus influenzae, and hepatitis B virus as well as antiretroviral therapy for asymptomatic persons . Quantitative measurement of viral titers appears promising for the monitoring of HIV disease and antiretroviral therapy; the correlations of these titers with clinical end points need to be confirmed. Clin Infect Dis, 1995 Apr, 20(4), 924 - 30 Failure of treatment for chancroid in Rwanda is not related to human immunodeficiency virus infection: in vitro resistance of Haemophilus ducreyi to trimethoprim-sulfamethoxazole; Bogaerts J et al.; A comparative open study was performed to evaluate the efficacy of single doses of ciprofloxacin (500 mg) and trimethoprim-sulfamethoxazole (TMP-SMZ; 640 mg/3,200 mg) for the treatment of culture-proven chancroid . Clinical cure or improvement was observed 7 days after treatment in 32 (76.2%) of the 42 patients who received ciprofloxacin and 21 (52.5%) of the 40 patients who received TMP-SMZ (P = .04) . Cultures for one (4.5%) of 22 patients not cured with ciprofloxacin and 16 (59.3%) of 27 patients not cured with TMP-SMZ were still positive for Haemophilus ducreyi 7 days after treatment (P < .001) . Although 77 (71.3%) of the 108 patients tested were seropositive for HIV-1 antibody, HIV infection and the degree of CD4+ lymphocyte depletion had no effect on clinical and bacteriologic outcome . All isolates of H . ducreyi were highly susceptible to ciprofloxacin (MIC, 0.004-0.06 mg/L) . In contrast, resistance to TMP-SMZ (MIC, > or = 4/76 micrograms/mL) was observed in 48.9% of isolates (22 of 45) and was significantly associated with treatment failure . Therefore, the administration of TMP-SMZ, in single or multiple doses, is no longer indicated for the treatment of chancroid in Rwanda. Clin Infect Dis, 1995 Apr, 20(4), 854 - 60 Etiology of acute respiratory tract infections among children in a combined community and hospital study in Rio de Janeiro; Sutmoller F et al.; We reviewed data collected between January 1987 and December 1989 on the etiology of acute respiratory infections (ARI) among 827 children in two low-income communities and a hospital in Rio de Janeiro . Respiratory syncytial virus was identified in 38% of cases of ARI, influenza A virus in 1%, parainfluenza 3 virus in 1%, and multiple viruses in 1% . Respiratory syncytial virus was most prevalent among hospitalized children, with seasonal increases in the late fall and winter . The principal bacterial pathogens were Staphylococcus aureus, gram-negative bacteria, Streptococcus pneumoniae, and alpha-hemolytic streptococci . Specimens that were most often positive were pleural fluid (46%) and specimens from other normally sterile sites (24%); normally sterile sites included the CSF, trachea, and lungs . Urine counterimmunoelectrophoresis for S . pneumoniae and Haemophilus influenzae polysaccharide antigens was positive in 3% and 2% of cases, respectively . Pharyngeal cultures yielded low numbers of S . pneumoniae and H . influenzae organisms and higher numbers of gram-negative bacteria . This study demonstrates the high incidence of ARI (4.5 cases per 100 child-weeks) among children in Rio de Janeiro and the high morbidity associated with the illness (ARI is responsible for 25%-50% of all pediatric hospitalizations) and the fact that continued attention must be paid to both viral and bacterial agents of ARI. J Paediatr Child Health, 1995 Apr, 31(2), 99 - 104 Clinical manifestations and outcome of Haemophilus influenzae type b disease; Gilbert GL et al.; OBJECTIVE: To document clinical manifestations, laboratory findings and outcome of childhood Haemophilus influenzae type b (Hib) infections . METHODOLOGY: Medical records of 235 children with Hib disease admitted to hospital during a 2 year period were reviewed; additional information was obtained by questionnaire and follow up 6 weeks after discharge . RESULTS: Three-quarters of patients presented with either meningitis or epiglottitis . Children with epiglottitis were older, had shorter illnesses and were less likely to have had antibiotics before admission than those with meningitis; 38% of the latter had been given some antibiotic therapy, with no apparent effect on the outcome . Fever persisted for 7 days or more in 23% of patients with meningitis . Death from meningitis occurred in 3.8% of patients and was due to fulminating disease . CONCLUSIONS: These data will assist in recognition and appropriate management of Hib disease as the clinical manifestations become less familiar following the introduction of immunization . Specific laboratory diagnosis is required for accurate surveillance, which should be maintained in order to ensure high immunization rates. Pediatr Infect Dis J, 1995 Apr, 14(4 Suppl), S23 - 6 Otitis media complications and treatment failures: implications of pneumococcal resistance; Poole MD; Classic complications of untreated otitis media include meningitis, lateral sinus thrombosis and chronic suppurative otitis media . In the past, in countries where otitis media is usually treated, complications have been rare, because of the good activity of almost all orally administered antibiotics against the most common cause of complications, Streptococcus pneumoniae . Treatment failures were usually caused by beta-lactamase-producing nontypable Haemophilus influenzae or by Moraxella (Branhamella) catarrhalis and were rarely associated with serious systemic infections . With the advent of multidrug-resistant pneumococci, however, serious and fatal infections can occur in the face of our most potent antimicrobial agents . The consequences of the emergence of multidrug-resistant pneumococci are likely to include more persistent purulent otitis media, increased usage of broad-spectrum antibiotics, an increase in surgical treatment rates for otitis media and, eventually, an increase in suppurative complications of otitis media . Medical treatment failures probably already surpass eustachian tube dysfunction as the most common reason for tympanostomy tube insertion . Multidrug-resistant pneumococci may be expected to change the way in which primary and secondary care is currently administered. Pediatr Infect Dis J, 1995 Apr, 14(4 Suppl), S19 - 22 Multicenter trial of cefpodoxime proxetil vs . amoxicillin-clavulanate in acute lower respiratory tract infections in childhood . International Study Group; Klein M; Acute lower respiratory tract infections in children are a worldwide public health problem, with an estimated 4 million potentially preventable deaths every year . Until recently, penicillin and related drugs were the treatment of choice for empiric therapy of paediatric lower respiratory tract infections . However, concerns over the emergence of penicillin-resistant strains of Streptococcus pneumoniae and beta-lactamase-producing strains of Haemophilus influenzae and Moraxella catarrhalis have led physicians to turn increasingly towards alternatives, such as the third generation cephalosporins . The oral extended spectrum cephalosporin cefpodoxime proxetil is highly active against the bacterial pathogens commonly associated with childhood lower respiratory tract infections . In order to evaluate its clinical efficacy in children with acute febrile lower respiratory tract infections, an international, multicenter, comparative, randomized open study was conducted in children ages 3 months to 11.5 years . Of 348 cases enrolled, 234 were randomized to cefpodoxime proxetil (8 mg/kg/day twice daily) and 114 to amoxicilin/clavanulate (amoxicillin 40 mg/kg/day 3 times a day) . The duration of treatment was 10 days . Pretreatment diagnosis was pneumonia in 292 patients, bronchiolitis in 19 patients and acute bronchitis in 37 patients . Pathogens isolated from 59 cases included H . influenzae (47.5%), S . pneumoniae (23.7%), M . catarrhalis (11.9%) and Haemophilus parainfluenzae (6.8%) . Clinical efficacy was evaluable in 278 children at the end of treatment when 95.2% of patients in the cefpodoxime proxetil group and 96.7% of patients in the amoxicillin/clavanulate group showed a satisfactory clinical response (cured or improved) . The improvement was sustained at the follow-up visit, 10 to 20 days after completion of treatment.(ABSTRACT TRUNCATED AT 250 WORDS) Pediatr Infect Dis J, 1995 Apr, 14(4 Suppl), S12 - 8 Clinical experience with cefpodoxime proxetil in acute otitis media; Cohen R; Although it varies from country to country, there is a worrying worldwide increase in antibiotic resistance among pathogens causing otitis . This has led to a search for therapeutic alternatives to the reference treatment, which is still amoxicillin in many countries . Cefpodoxime proxetil is one such alternative . Six comparative randomized trials of cefpodoxime proxetil in childhood acute otitis media have been published or presented at international conferences . They involved a total of 1188 patients, 658 of whom received cefpodoxime proxetil and 530 of whom received the comparator drug (amoxicillin/clavulanic acid in 3 trials, cefaclor in 1, and cefixime in 2); duration of treatment varied from 5 days for cefpodoxime proxetil to 10 days for amoxicillin/clavulanic acid, and the age of the children included ranged from 2 months to 12 years . The clinical efficacy of cefpodoxime proxetil was at least equivalent to that of the comparators in 4 trials and significantly better in 2 trials . Firstly, in one study vs . amoxicillin/clavulanic acid, the superiority of cefpodoxime proxetil (8 mg/kg/day twice daily) in terms of healing at the end of treatment and in terms of the number of normal tympanograms at the follow-up visit was shown . Secondly, in a study performed by our group, vs . cefixime, cefpodoxime proxetil (8 mg/kg/day twice daily) showed a better healing rate at the end of treatment in febrile and painful acute otitis media . The microbiologic and pharmacokinetic data show that cefpodoxime proxetil is one of the most active compounds against Haemophilus influenzae and Streptococcus pneumoniae.(ABSTRACT TRUNCATED AT 250 WORDS) J Clin Microbiol, 1995 Apr, 33(4), 1036 - 8 Alterations in sample preparation increase sensitivity of PCR assay for diagnosis of chancroid; Johnson SR et al.; A PCR assay for the detection of Haemophilus ducreyi in clinical specimens taken from genital ulcers was developed . Although H . ducreyi, when present in such specimens, could be detected by PCR, the sensitivity of the assay was reduced by the presence of Taq polymerase inhibitors in the specimen . The sensitivity of the PCR assay was improved by the use of detergents in preparing nuclei acids from clinical specimens and by the inclusion of a dialysis step prior to amplification . In addition, sodium phosphate included in the transport medium was found to be an inhibitor of the Taq polymerase. Antimicrob Agents Chemother, 1995 Apr, 39(4), 910 - 6 Antimicrobial activity of SM-17466, a novel carbapenem antibiotic with potent activity against methicillin-resistant Staphylococcus aureus; Sumita Y et al.; The in vitro and in vivo antibacterial activities of SM-17466, a new 1 beta-methyl carbapenem, were evaluated against a wide range of clinical bacterial isoaltes and compared with the activities of meropenem, imipenem, vancomycin, and arbekacin . SM-17466 had a broad spectrum of action against gram-positive bacteria, showing especially potent activity against methicillin-resistant staphylococci . The MICs of SM-17466, meropenem, imipenem, vancomycin, and arbekacin at which 90% of clinical isolates of methicillin-resistant Staphylococcus aureus were inhibited were 3.13, 50, 100, 1.56, and 3.13 micrograms/ml, respectively . This activity of SM-17466 was almost equivalent to those of the antibiotics used for the treatment of infections caused by this organism . SM-17466 also showed bactericidal activity against methicillin-resistant S . aureus . In contrast, SM-17466 was less active against gram-negative bacteria, especially against Pseudomonas aeruginosa, compared with the other carbapenems; however, of the carbapenems, SM-17466 exhibited the highest activity against Haemophilus influenzae and Bacteriodes fragilis . SM-17466, at a 50% inhibitory concentration of less than 1 microgram/ml, bound to penicillin-binding proteins 1 to 4 in methicillin-susceptible S . aureus and also had good binding to penicillin-binding protein 2' in a methicillin-resistant strain (50% inhibitory concentration, 5.9 micrograms/ml) . This high affinity, which was 10 and 20 times greater than those for meropenem and imipenem, respectively, was reflected in the potent activity of SM-17466 against methicillin-resistant S . aureus . SM-17466 demonstrated excellent in vivo efficacy against methicillin-susceptible and -resistant S . aureus strains in a mouse peritoneal infection model: the efficacy of SM-17466 against methicillin-resistant strains was equal to or one-third that of vancomycin . This activity was comparable to the in vitro activity of SM-17466 . The subcutaneous injection of SM-17466 in mice revealed that the half-life of SM-17466 in serum was about 18 min, intermediate between those of vancomycin and arbekacin and 1.5-fold that of imipenem-cilastatin . SM-17466 was resistant to hydrolysis by swine renal dehydropeptidase I, to an extent comparable to the resistance shown by meropenem. Nippon Jibiinkoka Gakkai Kaiho, 1995 Apr, 98(4), 659 - 68 {The effect of cefaclor and cefixime on nasopharyngeal pathogens in children}; Tomiyama M; Changes in nasopharyngeal flora were investigated in children with acute otitis media and with acute exacerbations of chronic sinusitis in whom antibiotic therapy of relatively long duration was required until substantial improvement in clinical findings was achieved . 1 . The antibiotics used were two cephalosporins, i.e., cefaclor (CCL) and cefixime (CFIX), administered to 18 patients each for 1 week and to 26 and 20 patients, respectively, for 2 weeks . Bacteriologic examination of the nasopharyngeal mucosa was performed at the first visit and at 1 week in those who underwent antibiotic therapy for 1 week, and at the first visit and at 1 and 2 weeks in those treated with antibiotics for 2 weeks . 2 . The elimination rates for the infecting microorganisms in the patients in the CCL-treated group were 30% for Haemophilus influenzae, 83% for Staphylococcus aureus, 100% for Streptococcus pyogenes and 100% for Streptococcus pneumoniae at 1 week, and 18% for H . influenzae, 100% for S . aureus and 100% for S . pyogenes at 2 weeks of antibiotic therapy . Replacement of S . aureus and S . pyogenes by H . influenzae was observed . 3 . The elimination rates for infecting bacteria in the patients in the CFIX-treated groups were 61% for H . influenzae, 50% for S . aureus, 75% for S . pyogenes, 80% for S . pneumoniae and 100% for Moraxella catarrhalis at 1 week, and 72% for H . influenzae, 0% for S . aureus, 100% for S . pyogenes, and 0% for S . pneumoniae at 2 weeks of antibiotic therapy . The elimination rate for H . influenzae at 2 weeks was significantly higher than the corresponding value for the CCL-treated group . Replacement of H . influenzae by S . aureus and S . pneumoniae and of S . pyogenes by S . aureus was detected . 4 . There was one patient with acute otitis media in the CFIX-treated group in whom a clinical relapse occurred due to H . influenzae persisters in the nasopharynx . Thus the diagnosis in this patient was so-called "recurrent otitis media" . 5 . H . influenzae tended to persist after exposure to therapeutically adequate concentrations of CCL, as did S . aureus and S . pneumoniae following treatment with CFIX . Thus, it would seem that ample heed must be given to persistence, particularly of H . influenzae and S . pneumoniae, the most common causative agents of acute otitis media in childhood . 6 . A significant rise in the MICs of the cephalosporins was observed in 4 of 43 patients in whom the same type of organism was isolated from the nasopharynx at weekly intervals during antibiotic therapy.(ABSTRACT TRUNCATED AT 400 WORDS) Pediatrics, 1995 Apr, 95(4), 522 - 7 Safety and immunogenicity of PRP-T combined with DTP: excretion of capsular polysaccharide and antibody response in the immediate post-vaccination period; Miller MA et al.; OBJECTIVE . To evaluate whether combining Haemophilus influenzae type b capsular polysaccharide covalently linked to tetanus toxoid (PRP-T) and diphtheria-tetanus-pertussis (DTP) in one syringe produced a vaccine that was safe and immunogenic . DESIGN . Randomized clinical trial . SETTING . Suburban New Orleans pediatric population . PARTICIPANTS . Convenience sample of 150 healthy infants . METHODS . Enrollees were randomized to receive DTP and PRP-T in one injection (Group 1), DTP and PRP-T separately (Group 2), or DTP and H influenzae type b capsular saccharide coupled to a nontoxic variant of diphtheria toxin, CRM197 (HbOC) separately (Group 3) at 2, 4, and 6 months of age . All infants received oral polio vaccine at 2 and 4 months of age . Parents were instructed to record side effects on a standardized form after each vaccine administration . Blood was drawn before each immunization and at 7 months of age; an additional blood and a urine specimen was obtained 2 to 3 days after one of the vaccination visits . Serum was assayed for H influenzae anticapsular antibody (anti-PRP), anti-pertussis toxoid, anti-fimbrial hemagglutinins, anti-diphtheria and anti-tetanus toxoid antibodies, and antibody to polio viruses . Urine was assayed for H influenzae type b capsular polysaccharide . RESULTS . The rate of occurrence of fever did not differ significantly between groups . Local swelling and erythema occurred more often at the administration site in Group 1 infants than at the DTP administration sites of infants in Groups 2 and 3 after the first and second vaccinations . The mean concentration of all antibodies we assayed did not differ significantly when Group 1 and 2 infants were compared . HbOC recipients (Group 3) had lower mean anti-H influenzae anticapsular antibody and higher mean anti-diphtheria and anti-tetanus antibody concentrations after two and three doses compared with Group 1 and Group 2 infants . No group had a significant change in mean anti-PRP antibody concentration 2 to 3 days after vaccination with any dose . After vaccination, antigenuria occurred less frequently in Group 1 infants (54%, 78%, and 72% in Groups 1, 2, and 3, respectively, P < .01) . CONCLUSIONS . Combining PRP-T and DTP produced a combination vaccine associated with a slight increase in the rate of erythema and swelling but with similar immunogenicity of the vaccine components and oral polio vaccine. Clin Exp Immunol, 1995 Apr, 100(1), 47 - 53 Severity of infections in IgA deficiency: correlation with decreased serum antibodies to pneumococcal polysaccharides and decreased serum IgG2 and/or IgG4; French MA et al.; In order to define abnormalities of humoral immunity which determine susceptibility to respiratory tract infections in IgA-deficient adults, serum IgG subclass concentrations, and serum concentrations of pneumococcal antibodies and Haemophilus influenzae type B (Hib) antibodies sera from IgA-deficient adults with and without susceptibility to respiratory tract infections were compared . Infection susceptibility was not related to the degree of IgA deficiency, but was related to deficiency of IgG4 and, to a lesser extent, IgG2, as well as to low basal serum concentrations of pneumococcal polysaccharide antibodies . The combination of IgG2 and/or IgG4 deficiency and a non-protective basal serum concentration of antibody against two or more pneumococcal polysaccharides was present in the serum of six of 12 (50%) patients with severe infections, but only one of 44 (2%) patients without infections . Furthermore, the preservation of antibody responses against the most immunogenic pneumococcal polysaccharide type 3, but not against the less immunogenic types 7F, 9N and 14, in patients with severe infections suggested that abnormalities of pneumococcal polysaccharide antibody responses might include defects of affinity maturation. Avian Dis, 1995 Apr-Jun, 39(2), 304 - 8 Phenotypic and molecular characterization of V-factor (NAD)-independent haemophilus paragallinarum; Miflin JK et al.; In South Africa from early 1989 onward, strains of Haemophilus paragallinarum not requiring nicotinamide adenine dinucleotide (NAD) have been isolated from commercial chickens suffering from infectious coryza . Fifteen of these field isolates were characterized by biochemical typing, serotyping, restriction endonuclease analysis (REA), and ribotyping . The chosen isolates represented diversity in geographi