JAMA, 1994 Oct 12, 272(14), 1116 - 21 Effect of carrier protein priming on antibody responses to Haemophilus influenzae type b conjugate vaccines in infants; Granoff DM et al.; OBJECTIVE--To assess the effect of priming with diphtheria and tetanus toxoid vaccine (DT) at 1 month of age on the anticapsular polyribosylribitol phosphate (PRP) antibody responses of infants vaccinated with Haemophilus influenzae type b polysaccharide-tetanus toxoid conjugate (PRP-T) or PRP oligosaccharide-cross-reactive mutant diphtheria toxin conjugate (HbOC) . DESIGN--Randomized controlled trial with serum samples assayed blindly . PARTICIPANTS AND SETTING--Healthy infants enrolled in private pediatric practices; 94 (91%) of 103 infants had prevaccination and postvaccination serum samples available for analysis . INTERVENTIONS--Two groups received DT vaccination at 1 month of age and subsequent injections of PRP-T or HbOC conjugate vaccines at 2, 4, and 6 months of age . The control groups were not vaccinated with DT but received PRP-T or HbOC at the same ages as the carrier-primed groups . Infants in all groups were given a booster injection of unconjugated PRP at 12 months of age to assess induction of immunologic memory . MAIN OUTCOME MEASURE--Concentrations of serum antibody to PRP . MAIN RESULTS--The DT-primed infants given PRP-T had twofold to threefold higher geometric mean anti-PRP antibody responses after one (P < or = .01), two (P < or = .01), or three (P = .06) doses of conjugate vaccine than the infants of the unprimed group . The primed infants also had threefold higher memory antibody responses to the booster PRP injection given at 12 months of age (concentration of 24.4 vs 8.4 micrograms/mL in infants not primed with DT; P < .01) . The DT-primed infants given HbOC had twofold to threefold higher antibody responses after one (P = .07) or two (P < .01) doses of conjugate vaccine than the unprimed HbOC group, but there were no significant differences after the third dose of conjugate vaccine or after the PRP booster injection . CONCLUSIONS--Vaccination with DT at 1 month of age increases the magnitude of the anti-PRP antibody responses to conjugate vaccination . With HbOC, the effect of carrier priming was present for up to 6 months of age, whereas in infants vaccinated with PRP-T, enhanced immunity was present for at least 12 months. Presse Med, 1994 Oct 8, 23(30), 1376 - 80 {Oropharyngeal flora . Epidemiologic survey of prevalence}; Bergogne-Berezin E et al.; OBJECTIVES: Epidemiological surveys which are not frequently carried out in medical practice should provide useful information for the choice of antibiotics to be prescribed in community-acquired infections particularly with the recent development of therapeutic difficulties due to resistant strains . We therefore analyzed the prevalent pharyngeal flora in a general patient population . METHODS: The study was conducted during a single 24-hour period in 1991 by 43 general practitioners and included 645 subjects consulting for benign affections . No patient selection was made . Two pharyngeal swabs were obtained from each subject and cultured in aerobic and anaerobic conditions . Internationally accepted methods for identifying bacteria in pharyngeal samples all performed by one well-equipped laboratory . Beta-lactamase activity was determined with the nitrocephine technique, both directly and after culture . RESULTS: Patient age varied from 16 to 45 years; most (68.5%) consulted for reasons other than ear-nose-throat affections . Only 41 patients (4.3%) consulted for sore throat and 65.4% had not received antibiotics for at least 6 months . Haemophilus influenzae was found in 59.6% of the patients, 20% of the strains were beta-lactamase producers as were 83.7% of the Moraxella catarrhalis strains identified . CONCLUSION: These factors are indicators of potential risk of therapeutic failure when using beta-lactams unstable to beta-lactamases for the treatment of pharyngeal infections. MMWR Morb Mortal Wkly Rep, 1994 Oct 7, 43(39), 718 - 20 Update: childhood vaccine-preventable diseases--United States, 1994; Vaccination coverage of 2-year-old children--United States et al.; The primary goal of the Childhood Immunization Initiative (CII) is to increase, by 1996, vaccination levels for 2-year-old children to at least 90% for the most critical doses in the vaccination series (i.e., one dose of measles-mumps-rubella vaccine {MMR} and at least three doses each of diphtheria and tetanus toxoids and pertussis vaccine {DTP}, oral poliovirus vaccine, and Haemophilus influenzae type b vaccine {Hib}) and to at least 70% for three or more doses of hepatitis B (Hep B) vaccine (1) . This report presents estimates, based on the National Health Interview Survey (NHIS), of the annual national vaccination coverage levels for children aged 19-35 months (median: 27 months) for 1993, compares estimates for 1993 with those for 1992, and compares estimates for the first 6 months of 1993 with third and fourth quarter 1993 estimates. Pharmacoeconomics, 1994 Nov, 6(5), 442 - 52 Cost of treatment and prevention of Haemophilus influenzae type b disease . An international perspective; Clements DA; On the basis of immunogenicity and protective efficacy studies, 4 Haemophilus influenzae type b (Hib) conjugate vaccines have been licensed for administration to infants and children . Population based studies of Hib disease from Australia, Chile, Finland, Gambia, Israel, Switzerland, UK and US show that the relative and absolute incidence of Hib disease varies significantly . These differences in Hib epidemiology, and associated sequelae and hospital costs affect the cost-benefit analysis of preventive vaccination, necessitating unique calculations for each country . Published papers on the cost of Hib disease and the cost-benefit relationship of Hib vaccination have been based primarily on reports from the US, but more recently also on studies from Australia, Finland, Israel, Switzerland, UK, Sweden and Chile . All studies to date have produced favourable cost-benefit ratios . The implementation of Hib vaccination has led to the virtual disappearance of Hib disease in some of these countries . The lessons gained from these analyses are instructive not only for better understanding of the epidemiology of Hib disease, but also as a template for assessing the cost-benefit ratio of the implementation of preventive vaccination for other diseases. Control Clin Trials, 1994 Oct, 15(5), 360 - 78 Decision making during a phase III randomized controlled trial; Berry DA et al.; Experiments such as clinical trials should be carried out with specific objectives . For example, in a trial designed to prevent disease, specific considerations should be made concerning the impact of the trial on the health of the target population, including the participants in the trial . These objectives should be assessed continually in light of data accumulating from the trial . Accumulating evidence should be judged in the context of changing circumstances external to the trial, and the trial's design possibly modified . An important type of modification is stopping the trial . This is a sequential decision problem that can be addressed using a Bayesian approach and the methods of dynamic programming . As an example we consider a vaccine trial for the prevention of haemophilus influenzae type b . The objective we consider is minimizing the number of cases of this disease in a Native American population over a specified horizon . We assess the prior probability distribution of vaccine efficacy . We also assess the probability of regulatory approval for widespread use of the vaccine, depending on the data presented to the regulatory officials . In deciding whether to continue the trial we weigh the impact of the possible future results by their (predictive) probabilities . We address the sensitivity of the optimal stopping policy to the prior probability distribution, to the assessed probability of regulatory approval, and to the horizon. Thorax, 1994 Oct, 49(10), 999 - 1001 Penetration of amoxycillin/clavulanic acid into bronchial mucosa with different dosing regimens; Gould IM et al.; BACKGROUND--The efficacy of an antibiotic is related to its concentration at the site of infection . Previous studies of the concentrations of amoxycillin and clavulanic acid (co-amoxiclav) in respiratory secretions or whole lung tissue have suffered from methodological problems . The concentration of amoxycillin and clavulanic acid was determined in bronchial mucosal biopsy samples obtained at bronchoscopy following five different dosing regimens . METHODS--Bronchial biopsy and serum samples were obtained from 50 patients undergoing diagnostic bronchoscopy . Ten patients each received 375 mg, 625 mg, 750 mg, and 3.25 g oral, and 1.2 g intravenous co-amoxiclav 1-3 hours before bronchoscopy . The concentrations of clavulanic acid and amoxycillin were determined by high performance liquid chromatography using a microbore column, solid phase extraction, and preconcentration to improve sensitivity tenfold over previous methods . RESULTS--Concentrations of both clavulanic acid and amoxycillin in bronchial mucosa were dose related and were well above the MIC90 of co-amoxiclav for the common bacterial respiratory pathogens including Haemophilus influenzae, Micrococcus catarrhalis and Streptococcus pneumoniae for all dosing regimens . Mean mucosal levels were 200% and 118% of the corresponding serum levels for amoxycillin and clavulanic acid respectively . CONCLUSIONS--Amoxycillin and clavulanic acid are concentrated in bronchial mucosa and, even at the lowest dose of 375 mg orally, are likely to produce tissue levels in the lung sufficient to inhibit all the common community acquired respiratory pathogens. Pediatrics, 1994 Oct, 94(4 Pt 1), 517 - 23 Immunization practices of pediatricians and family physicians in the United States; Szilagyi PG et al.; OBJECTIVE . To assess current practices and attitudes among pediatricians and family physicians across the United States regarding immunizations . DESIGN . Survey of a random sample of pediatricians and family physicians . SUBJECTS . Fellows of the American Academy of Pediatrics (N = 746) and American Academy of Family Medicine (N = 429) . SURVEY TOPICS . General immunization practices (eg, types of visits during which vaccinations are provided, mechanisms to identify undervaccinated children); and opinions about perceived barriers to immunizations, acceptance of alternative sites for immunizations, and possible immunization requirements for Medicaid and The Special Supplemental Food Program for Women, Infants, and Children (WIC) . RESULTS . Pediatricians and family physicians (combined) reported the following: immunizing children during acute illness visits (28%), follow-up visits (90%), and chronic illness visits (77%); using computer or reminder files to identify undervaccinated children (13%); and simultaneously administering four vaccines (diphtheria-tetanus-pertussis, oral poliovaccine, measles, mumps, and rubella and Haemophilus influenzae type b) to an eligible 18-month-old child (66%) . Physicians perceived the following as barriers to immunizations: missed preventive visits (40%), vaccine costs (24%), lack of insurance coverage (24%), inability to track undervaccinated patients (22%), incomplete immunization records (12%), and missed vaccination opportunities (12%) . Physicians agreed with offering vaccinations during hospitalizations (51%) or emergency department visits (30%), and with immunization requirements for continued eligibility for Medicaid (66%) or WIC (64%) . Pediatricians were more likely to vaccinate during chronic illness and follow-up visits, and were more likely to use systems to track undervaccinated children (P < .05); however, most immunization practices and attitudes of pediatricians and family physicians were similar . Physicians who graduated from medical school more recently and those in high-risk urban practices were more likely to vaccinate during acute illness visits, provide simultaneous vaccinations, and favor vaccinations in hospital settings . CONCLUSIONS . Vaccination rates might be improved by closer adherence to current immunization guidelines regarding vaccinations during all encounters and simultaneous vaccinations, by developing systems to identify undervaccinated children, and by reducing patient costs for vaccinations . Current immunization practices fall short of the immunization guidelines; changes in individual practice styles will be required to conform with these standards. Otolaryngol Head Neck Surg, 1994 Oct, 111(4), 513 - 8 Tobacco smoke and otitis media in the chinchilla model; Antonelli PJ et al.; To determine whether tobacco smoke contributes to the pathogenesis of acute otitis media, chinchillas were exposed to mainstream tobacco smoke or sham conditions (cigarettes not lit) in a Walton smoke exposure machine for 20-minute cycles two or three times daily . After 6 to 8 weeks of daily exposure, 12 chinchillas were nasally injected with Streptococcus pneumoniae, and 18 chinchillas were injected into both middle ears with nontypable Haemophilus influenzae . Smoke or sham exposures were continued for 2 to 4 weeks after injection . Otitis media developed in none of the 12 nasally injected chinchillas and in all 18 chinchillas whose middle ears were injected with nontypable Haemophilus influenzae . Persistence of middle ear effusion and persistence of nontypable Haemophilus influenzae in the middle ear effusion were not different between the smoke- and sham-exposed groups . This suggests that mainstream smoke exposure does not change the natural course of otitis media in the chinchilla model. J Pediatr, 1994 Oct, 125(4), 581 - 4 Response of immunocompromised children with solid tumors to a conjugate vaccine for Haemophilus influenzae type b; Shenep JL et al.; Serum antibody response to a conjugated Haemophilus influenzae type b polyribosylribitol phosphate-diphtheria toxoid vaccine was assessed in nonvaccinated children aged 1 1/2 to 5 years receiving chemotherapy for solid tumors . Responses occurred in 21 (42%) of 50 children after first vaccination, and in 10 (45%) of 22 revaccinated children. J Pediatr, 1994 Oct, 125(4), 571 - 6 Postlicensure effectiveness of the Haemophilus influenzae type b polysaccharide-Neisseria meningitidis outer-membrane protein complex conjugate vaccine among Navajo children; Harrison LH et al.; The incidence of invasive Haemophilus influenzae type b (Hib) infection was decreased significantly among Navajo children since the licensure of Hib conjugate vaccines, even though two lots of Hib (polyribosylribitol phosphate)-meningococcal B outer-membrane protein conjugate vaccine (PRP-OMP) widely used among the Navajo were later found to be of low immunogenicity . We measured the effectiveness of all Hib conjugate vaccines combined, PRP-OMP alone, and the PRP-OMP lots with lower-than-expected immunogenicity among Navajo infants and children . This was a matched case-control study using active, laboratory-based surveillance for the ascertainment of Navajo children 2 1/2 to 59 months of age with invasive Hib infection; 45 patients with infection and 180 control subjects were enrolled . The effectiveness of one, two, and three doses, respectively, of all Hib conjugate vaccines combined was 96% (95% confidence interval (CI) 65%, 99%), 99% (95% CI, 69%, 100%), and 99% (95% CI - 57%, 100%) . The effectiveness of one or more doses of PRP-OMP was 95% (95% CI, 66%, 99%) . The effectiveness of a single dose of the lots of lower-than-expected immunogenicity was 89% (95% CI, -8%, 99%) . The Hib conjugate vaccine coverage increased from 49% during 1991 to 94% during 1992; no control subjects younger than 18 months of age were enrolled during 1993 . The occurrence of invasive Hib infections in this population after licensure of Hib conjugate vaccines was the result of gradual vaccine uptake, not poor vaccine effectiveness . The use of PRP-OMP has been highly effective despite concerns about the immunogenicity of several lots. J Infect Dis, 1994 Oct, 170(4), 862 - 6 Nasopharyngeal colonization with nontypeable Haemophilus influenzae and recurrent otitis media . Tonawanda/Williamsville Pediatrics; Harabuchi Y et al.; The relationship between nasopharyngeal colonization with nontypeable H . influenzae and recurrent otitis media was assessed in 157 children followed prospectively from birth through 12 months of age . Forty-nine (31%) became colonized . Nasopharyngeal secretory IgA (sIgA) reactive with the P6 outer membrane protein was detected in all colonized children . Reduction or elimination of the organism was associated with a better mucosal immune response (560 +/- 864 units/ng/mL of sIgA) than was persistence in the nasopharynx (121 +/- 81; P = .04) . Forty colonized children (82%) and 61 noncolonized children (56%) developed otitis media (P = .004); colonized children were four times more likely to be classified as otitis prone (P = .003) . The frequency of otitis media episodes was directly related to the frequency of colonization (r = .42, P < .01) . These results demonstrate a strong relationship between nasopharyngeal colonization patterns and otitis media . The mucosal immune response may be important in elimination of potential pathogens from the respiratory tract. Infect Immun, 1994 Oct, 62(10), 4515 - 25 Identification of a locus involved in the utilization of iron by Haemophilus influenzae; Sanders JD et al.; Haemophilus influenzae has an absolute requirement for heme for aerobic growth . This organism can satisfy this requirement by synthesizing heme from iron and protoporphyrin IX (PPIX) . H . influenzae type b (Hib) strain DL42 was found to be unable to form single colonies when grown on a medium containing free iron and PPIX in place of heme . In contrast, the nontypeable H . influenzae (NTHI) strain TN106 grew readily on the same medium . A genomic library from NTHI strain TN106 was used to transform Hib strain DL42, and recombinants were selected on a medium containing iron and PPIX in place of heme . A recombinant plasmid with an 11.5-kb NTHI DNA insert was shown to confer on Hib strain DL42 the ability to grow on iron and PPIX . Nucleotide sequence analysis revealed that this NTHI DNA insert contained three genes, designated hitA, hitB, and hitC, which encoded products similar to the SfuABC proteins of Serratia marcescens, which have been shown to constitute a periplasmic binding protein-dependent iron transport system in this enteric organism . The NTHI HitA protein also was 69% identical to the ferric-binding protein of Neisseria gonorrhoeae . Inactivation of the cloned NTHI hitC gene by insertion of an antibiotic resistance cartridge eliminated the ability of the recombinant plasmid to complement the growth deficiency of Hib DL42 . Construction of an isogenic NTHI TN106 mutant lacking a functional hitC gene revealed that this mutation prevented this strain from growing on a medium containing iron and PPIX in place of heme . This NTHI hitC mutant was also unable to utilize either iron bound to transferrin or iron chelates . These results suggest that the products encoded by the hitABC genes are essential for the utilization of iron by NTHI. Infect Immun, 1994 Oct, 62(10), 4460 - 8 Localization of high-molecular-weight adhesion proteins of nontypeable Haemophilus influenzae by immunoelectron microscopy; Bakaletz LO et al.; A family of high-molecular-weight (HMW) surface-exposed proteins important in the attachment of nontypeable Haemophilus influenzae (NTHi) to human epithelial cells was previously identified (J . W . St . Geme III, S . Falkow, and S . J . Barenkamp, Proc . Natl . Acad . Sci . USA 90:2875-2879, 1993) . In the present investigation, indirect immunogold labeling and electron microscopy were used to localize these proteins on three clinical isolates of NTHi, mutants deficient in expression of one or both HMW proteins, and embedded sections of human oropharyngeal cells after incubation with NTHi strain 12 . The filamentous material comprising the proteins was labeled with monoclonal antibodies directed against two prototype HMW proteins (HMW1 and HMW2) of prototype NTHi strain 12 . Gold labeling was observed as a cap or discrete aggregate off one pole or centrally along one long axis of the bacterial cell . Heavily labeled, non-bacterial-cell-associated, disk-like aggregates of the HMW proteins were frequently noted in both bacterial preparations as well as in association with the oropharyngeal cell surface and intracellularly . Mutants demonstrated diminished labeling or an absence thereof, respectively, which correlated well with their previously demonstrated reduced ability or inability to adhere to Chang conjunctival epithelial cells in vitro . The Haemophilus HMW proteins share antigenic determinants with and demonstrate amino acid sequence similarity to the filamentous hemagglutinin protein of Bordetella pertussis, a critical adhesin of that organism . The studies presented here demonstrate that the Haemophilus proteins and B . pertussis filamentous hemagglutinin show impressive morphologic and perhaps additional functional similarity. Eur J Clin Microbiol Infect Dis, 1994 Oct, 13(10), 857 - 65 Multi-investigator evaluation of the efficacy and safety of cefprozil, amoxicillin-clavulanate, cefixime and cefaclor in the treatment of acute otitis media; Kafetzis DA; Cefprozil was evaluated in the treatment of acute otitis media with effusion in three open, randomized, multicenter comparative clinical trials . In two trials, 891 pediatric patients were enrolled to either cefprozil or amoxicillin-clavulanate dosage regimens . The treatment groups were comparable in demographic characteristics, and presented with otalgia, middle-ear effusion, or inflamed or bulging tympanic membrane on otoscopic examination . In all patients, tympanocentesis and a culture were required . Two cefprozil oral doses were evaluated, 30 mg/kg/day and 40 mg/kg/day divided into two equal doses (b.i.d.) . Amoxicillin-clavulanate was administered at 40 mg/kg/day in three divided doses (t.i.d.) . The recommended duration of therapy was ten days . The predominant bacteria isolated were Haemophilus influenzae and Moraxella catarrhalis . The overall satisfactory clinical response rates were similar for cefprozil (83%) and amoxicillin-clavulanate (81%) . The bacteriological response rates did not differ significantly, at 84% and 82% . Cefprozil eradicated the most common pathogen, Streptococcus pneumoniae, more often at 91%, vs . 84% for amoxicillin-clavulanate . The eradication rates were similar against Haemophilus influenzae and Moraxella catarrhalis . The patients treated with cefprozil had a lower rate of adverse clinical events (11%) compared to those with amoxicillin-clavulanate (20%) . More gastrointestinal adverse experiences, including diarrhea, were reported in the amoxicillin-clavulanate-treated patients . In Study 3, cefprozil 30 mg/kg/day (b.i.d.) was compared to cefaclor 40 mg/kg/day (t.i.d.) and cefixime 8 mg/kg/day (q.d) in the treatment of acute otitis media in 388 pediatric patients . The patients were treated for 10 days, with a follow-up of 18 days . The overall clinical cure rates were 85%, 89% and 85%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) Mol Cell Probes, 1994 Oct, 8(5), 417 - 22 Single polymerase chain reaction for the detection of tetracycline-resistant determinants Tet K and Tet L; Pang Y et al.; Oligonucleotide primers were used in polymerase chain reaction assays to detect tetracycline-resistant determinants Tet K and Tet L . Forty-three isolates representing 11 genera carrying Tet K and/or Tet L determinants gave appropriate PCR products . The PCR products hybridized with labelled Tet K or Tet L probes, and differentiated the Tet K from the Tet L PCR products . We confirmed that two Haemophilus aphrophilus carried the Tet K determinant and two Veillonella parvula carried the Tet L determinant . This is the first time that either of these two genes have been found in Gram-negative species . Four vaginal samples, previously positive with the Tet M/O PCR assay, were also positive with the Tet K/L assay . This is the first report of a PCR assay for the detection of Tet K and Tet L determinants in bacteria or clinical specimens. Mil Med, 1994 Oct, 159(10), 629 - 31 Pneumonia in military recruits; Amundson DE et al.; Lower respiratory disease is a major source of morbidity in military recruits, with hospitalization rates for pneumonia more than 30 times that of the non-recruit population . The etiologic agent remains unknown in over 75% of cases . This study prospectively examined the etiology of pneumonia among recruits at Naval Training Center, San Diego, California . Recruits presenting with cough, fever, or shortness of breath and pulmonary infiltrates on chest X-ray were eligible for enrollment . A standardized scoring form and focused physical exam were completed on each subject . Sputum specimens were obtained for Gram's stain and culture, DNA probing for Legionella and Mycoplasma species, and direct fluorescent antibody staining for Legionella . Acute and convalescent serologies were performed for adenovirus, influenza A and B, Mycoplasma pneumoniae, Chlamydia group, and respiratory syncytial virus . Of 110 eligible patients, 100 consented to enrollment and 75 patients completed the study . Etiologic diagnoses were obtained in 40 of the patients (53%) . M . pneumoniae, Haemophilus influenzae, and viruses accounted for the majority of infections . Mixed infections were seen in six patients . Forty-seven percent of patients had no diagnosis established . Pneumonia in this series of military recruits was frequently caused by M . pneumoniae and H . influenzae . Fifty percent of cases were undiagnosed with routinely available laboratory methods . Further studies are warranted to more clearly define the etiologic agents of recruit pneumonia and the utility of prophylactic measures. J Vet Med Sci, 1994 Oct, 56(5), 1017 - 9 Characterization of Haemophilus paragallinarum isolated in the Philippines; Nagaoka K et al.; In 1989 and 1990, we isolated five strains of Haemophilus paragallinarum in the Philippines . Serological studies were done on three of these isolates . The results of haemagglutination, reactability to monoclonal antibodies and cross immunization with standard strains showed that they belonged to serotypes A, B and C respectively . This is the first report on the isolation of H . paragallinarum and on a related serological study in the Philippines. J Chemother, 1994 Oct, 6(5), 319 - 21 Incidence of lower respiratory tract infections caused by Mycoplasma, Chlamydia and Legionella: an Italian Multicenter Survey; Schito GC et al.; A collaborative retrospective study based on serologic diagnosis was conducted to assess the etiological role sustained by privileged pathogens in Italy . The results obtained indicate the Mycoplasma, Chlamydia and Legionella are important etiologic agents of lower respiratory tract infections in Italy since they account for about 31% of the cases taken into consideration in this survey . We found a high incidence of M . pneumoniae (12.3%), C . pneumoniae (10.5%) and L . pneumophila (8.3%) . These results are in line with similar figures reported in the recent literature . While the data gathered in our survey do not allow us to clarify the nature of the agents involved in the etiology of the majority (70%) of the respiratory infections occurring in Italy, it seems safe to assume that after Streptococcus pneumoniae and Haemophilus influenzae, the privileged pathogens represent the most common cause of lower respiratory tract infections. Pediatr Emerg Care, 1994 Oct, 10(5), 268 - 72 Delayed incubation of blood culture bottles: effect on recovery rate of Streptococcus pneumoniae and Haemophilus influenzae type B; Roback MG et al.; This study investigated the effects of incubation delay on the rate of recovery of common pediatric pathogens from blood culture bottles . Known concentrations of Streptococcus pneumoniae and Haemophilus influenzae type b (three isolates each) were inoculated into BACTEC NR-6A bottles with 1.0 mL of donor blood . Bottles were subjected to a time delay (zero to six hours) before incubation . The BACTEC NR-660 was used for incubation and measurement of positive conversion . Data were analyzed using chi 2 analysis, Fisher's exact test, logistic regression, and multiple logistic regression, with P < 0.05 considered significant . Immediate incubation yielded positive blood cultures in 88 of 100 and 65 of 70 bottles containing S . pneumoniae and H . influenzae type b, respectively, in the concentration range 1.0 to 9.99 colony-forming units per milliliter (CFU/ml) . For each organism, this was the minimal range required to produce a positive culture (P < 0.0001) . Bottles inoculated with 1 ml of blood containing organisms in the range of 1.0 to 9.99 CFU/ml were then subjected to incubation delay . The recovery rate of S . pneumoniae significantly (P = 0.0003) decreased from a two-hour delay (57 of 60; 95%) to a three-hour delay (42 of 60; 70%) . No significant change in recovery rate was seen in bottles inoculated with H . influenzae type b subjected to similar delays . Delayed incubation (two to six hours) of bottles inoculated with 1.0 ml of blood containing organisms in a concentration range of 1.0 to 9.99 CFU/ml of blood significantly decreases the recovery rate of S . pneumoniae but has no effect on H . influenzae type b. Pediatr Emerg Care, 1994 Oct, 10(5), 264 - 7 Occult bacteremia: is there a standard of care? Ros SP, Herman BE, Beissel TJ. The evaluation and management of patients with occult bacteremia is controversial . The purpose of this study was to define the prevailing practices in the emergency management of occult bacteremia . Short, anonymous surveys were mailed to all 517 members of the Section on Emergency Medicine at the American Academy of Pediatrics . Three hundred six (59%) of those surveyed returned completed questionnaires . Eleven different temperature cutoff points are used, and 105 (34%) consider occult bacteremia in patients with temperature above 39 degrees C . Seventeen different age intervals are used to define the patients at risk for occult bacteremia, and the age range three to 24 months is used by 173 (57%) of those surveyed . Complete blood cell count is the most commonly used screening test; it is routinely ordered by 225 respondents (74%) . One hundred thirty-seven participants (45%) routinely obtain blood cultures in all patients at risk for occult bacteremia, whereas 111 (36%) use the clinical appearance (toxicity) of the patient to determine whether a blood culture should be drawn . One hundred sixty-one (53%) of those surveyed routinely administer antibiotics to toxic-appearing patients pending the results of the blood culture . Laboratory criteria are used by 135 (44%) in the decision whether to administer empiric antibiotics . Ceftriaxone is the most commonly used antibiotic; it is routinely administered by 230 respondents (75%) . Twenty participants (7%) routinely admit all patients with Streptococcus pneumoniae, whereas 217 (71%) admit all patients with Haemophilus influenzae bacteremia and 234 (76%) admit all patients with Neisseria meningitidis bacteremia . We conclude that diversity exists in the evaluation and management of occult bacteremia. Antimicrob Agents Chemother, 1994 Oct, 38(10), 2419 - 25 In vitro activities of 12 orally administered antimicrobial agents against four species of bacterial respiratory pathogens from U.S . Medical Centers in 1992 and 1993; Barry AL et al.; Clinical isolates of Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus pyogenes, and Moraxella catarrhalis were gathered from 19 different clinical laboratories throughout the continental United States . The in vitro activities of 12 orally administered antimicrobial agents were compared by broth microdilution tests with 3,151 bacterial isolates . Among 890 H . influenzae isolates, 30% were capable of producing beta-lactamase enzymes (12 to 41% in different medical centers) . Most of the 619 beta-lactamase-negative H . influenzae strains were susceptible to ampicillicin (MIC, < or = 1.0 micrograms/ml): 5 strains were intermediate in susceptibility (MIC, 2.0 micrograms/ml) and 1 strain was ampilicillin resistant (MIC, 4.0 micrograms/ml) . Ninety-two percent of 698 M . catarrhalis strains were beta-lactamase positive . Of 799 S . pneumoniae isolates, 15% were intermediate in susceptibility to penicillin and 7% were resistant to penicillin . The prevalence of penicillin-susceptible pneumococci in different institutions ranged from 63 to 95% . Only 1% of 764 S . pyogenes isolates were resistant to the macrolides, but 5% of S . pneumoniae isolates were macrolide resistant . Only 71% of 58 penicillin-resistant S . pneumoniae isolates were erythromycin susceptible, whereas 97% of the 622 penicillin-susceptible strains were erythromycin susceptible . Penicillin-resistant pneumococci were also relatively resistant to the cephalosporins and amoxicillin . Penicillin-susceptible pneumococci were susceptible to amoxicillin-clavulanic acid (MIC for 90% of isolates tested {MIC90}, < or = 0.12/0.06 microgram/ml), cefixime (MIC90, 0.25 microgram/ml), cefuroxime axetil (MIC90, < or = 0.5 microgram/ml), cefprozil (MIC90, < or = 0.5 micrograms/ml), cefaclor (MIC90, 0.5 microgram/ml), and loracarbef (MIC90, 1.0 microgram/ml) . Most strains of the other species remained susceptible to the study drugs other than amoxicillin. Antimicrob Agents Chemother, 1994 Oct, 38(10), 2340 - 5 Evaluation of OPC-17116 against important pathogens that cause respiratory tract infections; Wakebe H et al.; The antibacterial activity of OPC-17116, a new fluoroquinolone antibacterial agent, against important pathogens that cause respiratory tract infections was evaluated in vitro and in vivo and compared with those of ciprofloxacin, ofloxacin, and norfloxacin . The pharmacokinetic profiles of OPC-17116 were studied in both mice and rats given the drug orally at doses of 50 and 40 mg/kg of body weight, respectively . OPC-17116 showed a high degree of distribution in the lung tissues of both species, with maximum concentrations of 29.6 and 32.0 micrograms/g, respectively . Furthermore, the drug concentrations in lung tissue were about 10 to 15 times greater than the concentrations in plasma . OPC-17116 showed potent antibacterial activity against such pathogens as Staphylococcus aureus, Streptococcus pneumoniae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Haemophilus influenzae, and Moraxella catarrhalis . The MICs of this compound for 90% of these organisms except methicillin-resistant S . aureus and P . aeruginosa ranged from < or = 0.006 to 0.78 microgram/ml . The in vitro antibacterial activity of OPC-17116 was reflected by the efficacy of a single oral dose against systemic bacterial infections in mice . OPC-17116 showed a superior effect against gram-positive bacteria, H . influenzae, and M . catarrhalis . In comparison with the other reference compounds, the efficacy of OPC-17116 was less than that of ciprofloxacin against K . pneumoniae and P . aeruginosa . OPC-17116 showed a greater therapeutic effect than the other drugs against experimental acute pneumonia caused by these organisms in mice or rats . This excellent therapeutic effect against respiratory tract infections may be a result of its high level of distribution in lung tissue. Clin Otolaryngol, 1994 Oct, 19(5), 441 - 5 Acute epiglottitis--aetiology, epidemiology and outcome in a population before large-scale Haemophilus influenzae type b vaccination; Hugosson S et al.; Over a period of 18 years 219 consecutive cases of acute epiglottitis were diagnosed and subsequently investigated in order to elucidate the aetiology, epidemiology and outcome of this disease in a well-defined population in Sweden before general vaccination against Haemophilus influenzae type b infection was introduced . Compared with the results from other parts of the industrialized world, high incidence rates were found in both children (14/100,000/year) and adults (2.3/100,000/year) . The annual trend showed a significant decline in incidence among children, whereas in adults it remained unchanged . In cases where the aetiological agent could be determined, infection with H . influenzae type b was the main cause of disease in all age groups . However, in adults 27% (6/22) had a disease caused by micro-organisms other than H . influenzae type b that were verified with a blood culture . Sixty-eight per cent had a negative blood culture . The mortality rate was 0.5% (1/219) and 6% (13/219) developed a significant complication of the disease. J Paediatr Child Health, 1994 Oct, 30(5), 393 - 7 Sequelae of Haemophilus influenzae type b meningitis in aboriginal and non-aboriginal children under 5 years of age; Bower C et al.; Between 1984 and 1990, 257 cases of Haemophilus influenzae type b (Hib) meningitis occurred in children under five years of age in Western Australia . We obtained information on possible sequelae in 131 cases (all non-Aboriginal) by medical record review and parental interview, and in a further 116 cases (60 non-Aboriginal, 56 Aboriginal) by medical record review only; no follow-up information was available for ten children (nine non-Aboriginal, 1 Aboriginal) . The incidence of Hib meningitis in children under five years of age was 26.3 per 100,000 for non-Aboriginal and 152.2 per 100,000 for Aboriginal children . The case fatality rate was 3.5% for non-Aboriginal children and 14.0% for Aboriginal children . Sequelae were recorded for 17.1% of non-Aboriginal and 22.4% of Aboriginal children who survived Hib meningitis . Surviving Aboriginal children experienced severe sequelae following Hib meningitis almost three times more frequently than surviving non-Aboriginal children (10.5% vs 3.6%), although mild and moderate sequelae were not more common in Aboriginal children . The information on incidence and severity of sequelae in this study was obtained by chart review and parental interview, and hence may be subject to error or bias, particularly for mild and moderate disabilities . Outcomes like death and severe sequelae, such as cerebral palsy and profound intellectual and physical disability, are less subject to bias . Of Aboriginal children who contracted Hib meningitis in Western Australia over the study period, 22.8% either died or had severe sequelae, while only 7.0% of non-Aboriginal children experienced these severe outcomes. Mol Microbiol, 1994 Oct, 14(2), 217 - 33 A Haemophilus influenzae IgA protease-like protein promotes intimate interaction with human epithelial cells; St Geme JW 3rd et al.; Haemophilus influenzae represents a common cause of human disease and an important source of morbidity and mortality . Disease caused by this organism begins with colonization of the upper respiratory tract . Several studies indicate that H . influenzae is capable of binding to and entering cultured human cells, properties which are potentially of relevance to the process of colonization . In the present study, we isolated an H . influenzae gene designated hap, which is associated with the capacity for in vitro attachment and entry . Analysis of the derived amino acid sequence of hap demonstrated significant homology with the serine-type IgA1 proteases expressed by H . influenzae and Neisseria gonorrhoeae . It is notable that the hap product shares the catalytic domain of the IgA1 proteases and appears to be processed and secreted in an analogous manner . We speculate that the hap gene product is an important determinant of colonization, perhaps enabling the organism to evade the local immune response and thereby persist within the respiratory tract. J Clin Microbiol, 1994 Oct, 32(10), 2441 - 7 Determination of parallelism and nonparallelism in bioassay dilution curves; Plikaytis BD et al.; There is a lack of consensus among investigators who use a variety of immunoassay techniques (e.g., enzyme-linked immunosorbent assay {ELISA} and radioimmunoassay) regarding the protocols for describing and forming standard reference or calibration curves and interpolating serum antibody concentrations . This confounds the issue of detecting the presence or absence of parallelism between standard reference serum and serially diluted serum sample curves . These curves must be parallel to support the assumption that the antibody-binding characteristics are similar enough to allow the determination of antibody levels in the diluted serum sample . There is no universal and widely adopted strategy for assessing parallelism in bioassays, and without an assurance of parallelism, investigators are not able to calculate reliable estimates for antibody concentrations in serum samples . Furthermore, single-point (dilution) serum assays do not provide information related to parallelism and nonparallelism, and this, too, may lead to considerable error when calculating antibody concentrations . When assay methodology, technique, and precision improve to the extent that standard reference serum and serially diluted serum sample curves are fit with little error, standard analysis of variance techniques are overly sensitive to negligible departures from parallelism . We present a series of guidelines that compose a protocol for assessing parallelism between bioassay dilution curves that are applicable to data derived from ELISAs . These criteria should be applicable, with minor modifications, to most immunoassay experimental situations and, most importantly, are not dependent on the mathematical model used to form the standard reference curve . These guidelines have evolved in our laboratories over the past 4 years during the performance of thousands of ELISAs for antibodies to the capsular polysaccharides of Neisseria meningitidis groups A and C and Haemophilus influenzae type b. J Clin Microbiol, 1994 Oct, 32(10), 2382 - 6 PCR for capsular typing of Haemophilus influenzae; Falla TJ et al.; A PCR method for the unequivocal assignment of Haemophilus influenzae capsular type (types a to f) was developed . PCR primers were designed from capsule type-specific DNA sequences cloned from the capsular gene cluster of each of the six capsular types . PCR product was amplified only from the capsular type for which the primers were designed . Product was confirmed by using either an internal oligonucleotide or restriction endonuclease digestion . A total of 172 H . influenzae strains of known capsular type (determined genetically) comprising all capsular types and noncapsulate strains were tested by PCR capsular typing . In all cases the PCR capsular type corresponded to the capsular genotype determined by restriction fragment length polymorphism analysis of the cap region . When used in conjunction with PCR primers derived from the capsular gene bexA, capsulate, noncapsulate, and capsule-deficient type b mutant strains could be differentiated . PCR capsular typing overcomes the problems of cross-reaction and autoagglutination associated with the serotyping of H . influenzae strains . The rapid and unequivocal capsular typing method that is described will be particularly important for typing invasive H . influenzae strains isolated from recipients of H . influenzae type b vaccine. Jpn J Antibiot, 1994 Oct, 47(10), 1379 - 400 {Antibacterial activities of new quinolones against fresh clinical isolates}; Deguchi K et al.; In order to investigate antibacterial activities of new quinolones (NQs) against a number of clinical isolates obtained in our laboratory during a period from February, 1993 to January, 1994, minimum inhibitory concentrations (MICs) were determined using most of the NQs available in the market as of December, 1993 . The obtained results are summarized as follows: 1 . Noticeable differences were observed among the antibacterial activities of 8 different NQs tested against Gram-positive bacteria, i.e., there were large differences in their MIC distributions . Some differences were also observed among different NQs in ratios of NQ-resistant strains among Staphylococcus spp . From these results, it seems necessary to further study tolerance mechanisms of these Gram-positive bacteria toward different NQs and also to examine possible differences in antibacterial activities among different NQs against Gram-positive bacteria in clinical settings . 2 . MIC distributions against Gram-negative bacteria were also different among the 8 NQs tested . Though elevated MICs were observed against NQ-resistant Gram-negative bacteria in many cases, and somewhat higher, though not exceedingly high, MIC values than those against NQ-sensitive bacteria were found in other cases, patterns of MIC values against different NQ-resistant Gram-negative bacteria were similar for all of the 8 NQs tested . This may explain the fact that most of NQ-resistant Gram-negative bacteria showed similar resistant patterns to the 8 NQs tested . 3 . Among the NQ-resistant bacteria, were found Haemophilus influenzae and Neisseria gonorrhoeae strains . Ratios of resistant strains were approximately 10% or lower for the former and approximately 20% for the latter . 4 . With MICs of ampicillin and cefaclor used as control, it appears that benzylpenicillin (PCG)-insensitive or PCG-resistant Streptococcus pneumoniae (PISP or PRSP) and CEPs-resistant Escherichia coli are increasing. Jpn J Antibiot, 1994 Oct, 47(10), 1369 - 78 {Antimicrobial activities of cefditoren against clinical isolates obtained from outpatients}; Deguchi K et al.; To examine the antimicrobial activity of cefditoren (CDTR) against strains clinically isolated from outpatients at this hospital from November, 1993 to February, 1994, the minimum inhibitory concentrations (MICs) were determined including those of the control drugs . The results were as follows: 1 . CDTR showed strong antimicrobial activities against Staphylococcus aureus subsp . aureus, Streptococcus pyogenes and Streptococcus pneumoniae . The MICs of CDTR against benzylpenicillin-insensitive or -resistant S . pneumoniae distributed in the lowest concentration range even compared to those of the control drugs . 2 . CDTR showed strong antimicrobial activities against Haemophilus influenzae, Moraxella subgenus Branhamella catarrhalis, Escherichia coli, and Klebsiella spp . The MIC of CDTR against CEPs-resistant E . coli was lower than those of most control drugs . 3 . Since the microbes described above the major pathogens for the community-acquired infections, CDTR will be effective against infectious diseases transmitted at outpatient visits. Clin Infect Dis, 1994 Oct, 19(4), 768 - 9 Haemophilus influenzae epididymo-orchitis and bacteremia in a man infected with the human immunodeficiency virus; Cross JT Jr et al.; Haemophilus influenzae is a major bacterial pathogen in patients infected with the human immunodeficiency virus (HIV), although most infections with this organism occur in the respiratory tract . We describe an adult with HIV infection who presented with epididymo-orchitis due to H . influenzae . Eleven prior cases of H . influenzae epididymo-orchitis have been published, but all of these cases occurred in pediatric patients . Little is known about the prevalence of genitourinary tract infections caused by H . influenzae among adults . H . influenzae is a relatively rare cause of bacteremia in adults, but the frequency of H . influenzae bacteremia has been increasing among the HIV-positive population. Microb Pathog, 1994 Oct, 17(4), 277 - 82 Mapping of a strain-specific bactericidal epitope to the surface-exposed loop 5 on the P2 porin protein of non-typeable Haemophilus influenzae; Yi K et al.; The P2 protein is the major outer-membrane protein of non-typeable Haemophilus influenzae (NTHI) and shows extreme heterogeneity among strains . Based on the analysis of antigenic structure, the P2 protein consists of eight potentially surface-exposed loops . Previous studies of monoclonal antibodies (mABs) to a single strain of NTHI showed that P2 contains potentially immunodominant epitopes in loop 5 of the molecule . The goal of the current work is to test the hypothesis that strain-specific and potentially immunodominant epitopes are located in loop 5 of P2 in other strains of NTHI as well . Gene fragments which encode peptides of loop 5 of strains 2019 and 5657 were cloned into an expression vector and subjected to immunoassays with mABs which recognize surface-exposed, bactericidal, strain-specific epitopes . Each mAB recognized loop 5 of the P2 protein of the homologous strain . Analysis of mutant clones with minor amino acid changes showed a loss of reactivity with the mABs . These observations indicate that loop 5 of the P2 molecule contains strain-specific, abundantly expressed surface-exposed epitopes . This further supports the hypothesis that loop 5 is an immunodominant region of the P2 molecule. Clin Infect Dis, 1994 Oct, 19(4), 677 - 83 Haemophilus parainfluenzae endocarditis: application of a molecular approach for identification of pathogenic bacterial species; Hamed KA et al.; Haemophilus parainfluenzae is both a human oropharyngeal commensal bacterium and a cause of serious invasive disease . The fastidious growth characteristics of this organism and the poor specificity of traditional methods for species identification are likely to have led to inaccuracies in the diagnosis of infections caused by H . parainfluenzae and related organisms . We report a case of H . parainfluenzae endocarditis in which confusion related to microbial identification was resolved by the analysis of 16S ribosomal RNA sequences . Rapid identification was facilitated by amplification of 16S ribosomal DNA directly from cultured cells with use of the polymerase chain reaction and by direct DNA sequence determination of the amplified product . This procedure is potentially useful for the identification of fastidious bacterial pathogens by reference laboratories. Endocrinology, 1994 Oct, 135(4), 1488 - 95 Role of the extracellular regions of the parathyroid hormone (PTH)/PTH-related peptide receptor in hormone binding; Lee C et al.; The PTH/PTH-related peptide receptor is a member of a newly discovered family of G-protein-coupled receptors . Strikingly conserved features among these receptors include the positioning of eight extracellular cysteines and several other residues that are located predominantly within the membrane-embedded region . Deletion mutants or receptors with point mutations of the highly conserved cysteine residues were transiently expressed in COS-7 cells to evaluate PTH binding and PTH-stimulated cAMP production . Deletion of residues 61-105, which are encoded by exon E2 in the PTH/PTH-related peptide receptor gene, did not affect receptor function . An epitope derived from Haemophilus influenza hemagglutinin was, therefore, introduced into this portion of most receptors to allow the independent assessment of cell surface expression . PTH binding capacity was not reduced by the deletion of residues 258-278 in the first extracellular loop . Receptors with deletion of either residues 31-47 in the amino-terminal extension or residues 431-440 in the third extracellular loop failed to bind PTH, although expression of the receptor on the cell surface was only marginally reduced . Most other receptor mutants, including those in which each of the six cysteines in the amino-terminus was replaced by serines, failed to be processed and/or expressed appropriately, whereas the substitution of cysteine-281 or -351 had a less severe effect . The combined replacement of both cysteines concomitantly increased PTH binding and cell surface expression, suggesting the formation of a disulfide bond between these two residues . Our data indicate that residues near the amino-terminus and within the third extracellular loop are necessary for ligand binding, whereas more than 25% of the receptor's extracellular region appears not to be involved. Arch Intern Med, 1994 Sep 26, 154(18), 2086 - 91 Bacterial bronchitis and bronchiectasis in human immunodeficiency virus infection; Verghese A et al.; BACKGROUND: Bacterial pneumonia and sinusitis are important causes of morbidity in patients with human immunodeficiency virus (HIV) infection . We noted an increased incidence of bacterial bronchitis and bronchiectasis in our patients with HIV infection . METHODS: This study was conducted on persons with HIV infection at a county hospital and clinic . Bronchiectasis was diagnosed by bronchogram and computed tomography in one patient and by computed tomography alone in two others . Bacterial bronchitis was defined by a Gram's stain showing an abundance of neutrophils with a predominance of one or more bacteria and by a confirmatory sputum culture . Bronchoscopy with broncho-alveolar lavage was performed in patients with bronchitis to eliminate other causes of bronchial inflammation . RESULTS: Eighteen episodes of bacterial bronchitis in 10 patients are described . The mean CD4 lymphocyte counts for these patients was 0.061 x 10(9)/L (range, 0.001 to 0.203 x 10(9)/L) . The most common pathogens in 18 episodes of bacterial bronchitis were Haemophilus influenzae and Streptococcus pneumoniae (five episodes each) and Pseudomonas aeruginosa (four episodes) . Response to antibiotic therapy was usually rewarding though recurrences were frequent . Three patients with well-defined bronchiectasis who appeared to have developed, or who became symptomatic during the course of, HIV infection are described . Their mean CD4 cell count was 0.03 x 10(9)/L (range, 0.024 to 0.037 x 10(9)/L) . Haemophilus influenzae, Staphylococcus aureus, Pseudomonas cepacia, and P aeruginosa were recovered from these patients; the P aeruginosa was a mucoid strain . CONCLUSIONS: Recurrent bacterial bronchitis should be added to the list of bacterial infections that occur with increased frequency with HIV infection . Repeated bacterial bronchitis may lead to bronchiectasis, which may be more common in HIV infection than generally appreciated. J Am Vet Med Assoc, 1994 Sep 15, 205(6), 874 - 7 Vesicular adenitis syndrome in beef bulls; Grotelueschen DM et al.; Vesicular adenitis syndrome was diagnosed in 69.3% (52/75) of a group of yearling bulls on breeding soundness examination . Association of Haemophilus somnus infection with vesicular adenitis syndrome was confirmed in these bulls by microbial culture of vesicular gland fluid or semen samples and serologic testing. Clin Exp Immunol, 1994 Sep, 97(3), 411 - 6 Binding of mannan-binding protein to various bacterial pathogens of meningitis; van Emmerik LC et al.; Mannan-binding protein (MBP), a calcium-dependent plasma lectin, may play a role in the innate defence against microorganisms . After binding to carbohydrate structures at the bacterial surface, MBP activates the classical pathway of the complement system . To investigate the binding capacity of MBP to various bacteria associated with meningitis, an assay was developed to study the binding of MBP to bacteria grown in a semisynthetic fluid culture medium . Salmonella montevideo (containing a mannose-rich lipopolysaccharide (LPS)), used as a positive control strain, showed binding of radiolabelled MBP at a level of 80% compared with binding of MBP to zymosan . Binding of labelled MBP to Salm . montevideo was time-dependent, temperature-dependent and saturable . The binding was inhibited by unlabelled MBP, by mannose and by N-acetyl-D-glucosamine . Among bacterial pathogens often found to cause meningitis, a wide range of MBP binding capacities could be determined . The encapsulated Neisseria meningitidis (representatives from 11 serogroups other than group A were included: n = 22), N . mucosa (n = 1), Haemophilus influenzae type b (n = 10) and Streptococcus agalactiae (n = 5) had a low MBP binding capacity of 21.7% (95% confidence interval (CI) 3.3-40.1%) . Escherichia coli K1 (n = 11), Strep . suis (n = 5), Strep . pneumoniae (n = 10) and N . meningitidis serogroup A (n = 2) showed intermediate MBP binding capacity of 58.4% (95% CI 40.0-76.8%) . A third group consisting of non-encapsulated Listeria monocytogenes (n = 11), non-encapsulated H . influenzae (n = 2), non-encapsulated N . meningitidis (n = 2), N . cinera (n = 1) and N . subflava (n = 1) strains had a high MBP binding capacity of 87.5% (95% CI 62.5-112.5%) . The majority of encapsulated pathogens causing bacterial meningitis seem to have a rather low MBP binding capacity. Clin Exp Immunol, 1994 Sep, 97(3), 396 - 402 Cytokines in nasopharyngeal secretions; evidence for defective IL-1 beta production in children with recurrent episodes of acute otitis media; Lindberg K et al.; The host-parasite relationship in the nasopharynx of young children with bacterial colonization and antigen uptake in the mucosa and lymphatic tissue provides an opportunity to investigate infectious/inflammatory processes and responses . IL-1 beta, IL-6 and tumour necrosis factor-alpha (TNF-alpha) were analysed in nasopharyngeal secretions and serum from children with or without recurrent episodes of acute otitis media, from healthy adults and adults with or without recurrent episodes of acute otitis media, from healthy adults and adults with hypogammaglobulinaemia or selective deficiency of IgG3 . Nasopharyngeal secretions generally contained substantial amounts of IL-1 beta, IL-6 and TNF-alpha . In contrast, IL-1 beta, IL-6 and TNF-alpha were not detectable in sera on the same occasion . Children were found to have higher levels of IL-1 beta, IL-6 and TNF-alpha than healthy adults and than adults with immunodeficiency . High levels of IL-1 beta were associated with low or undetectable levels of IL-6 and TNF-alpha, whereas the opposite pattern was seen in association with low levels of IL-1 beta . This was especially true for children with recurrent episodes of acute otitis media (RAOM) . In children with nasopharyngeal colonization with Haemophilus influenzae, significantly higher levels of IL-1 beta, IL-6 and TNF-alpha (P = 0.0001, respectively) were found compared with non-colonized children . Notably, the RAOM children exhibited significantly lower levels of IL-1 beta, IL-6, and TNF-alpha in nasopharyngeal secretions (P = 0.0001, 0.01 and 0.0001, respectively) than healthy children . These results demonstrate local production of inflammatory cytokines in nasopharynx, related to bacterial colonization, and suggest that children with RAOM are poor nasopharyngeal cytokine producers. Infect Immun, 1994 Sep, 62(9), 4028 - 33 High-molecular-weight proteins of nontypeable Haemophilus influenzae mediate bacterial adhesion to cellular proteoglycans; Noel GJ et al.; A family of high-molecular-weight (HMW) surface-exposed proteins of nontypeable Haemophilus influenzae (NT H . influenzae) mediated adherence of these organisms to human epithelium . To better understand the molecular basis for this adherence, the role of glycosaminoglycans (GAGs), substances commonly expressed on cell surfaces, was examined . Bacterial adherence to cells with specific deficiencies in GAG biosynthesis was measured . HMW protein-dependent bacterial adherence to normal cells was significantly greater than adherence to cells deficient in sulfated GAGs or to cells deficient in heparan sulfate but overexpressing chondroitin sulfate . Cells expressing undersulfated heparan sulfate exhibited intermediate levels of bacterial adherence . The addition of exogenous dextran sulfate or heparin inhibited over 70% of the adherence of NT H . influenzae to normal cells, whereas hyaluronic acid and chondroitin sulfate tested at the same concentration (100 micrograms/ml) inhibited bacterial adherence by less than 11% . Treatment of cells with heparinase significantly reduced bacterial adherence . Following electrophoretic separation, HMW proteins were shown to bind directly to radiolabeled heparin . These results indicate that HMW protein-dependent adherence of NT H . influenzae is mediated by cellular sulfated GAGs and that heparan sulfate may be the predominant GAG involved in this process . However, the decreased adherence of bacteria to cells expressing undersulfated heparan sulfate and the inhibition of bacterial adherence by the addition of exogenous dextran sulfate suggest that bacterial adhesion to mammalian cells is likely to be influenced by a variety of factors, including the degree of sulfation and the specificity of the carbohydrate moieties contained in the cellular proteoglycans. Infect Immun, 1994 Sep, 62(9), 3881 - 9 The HMW1 adhesin of nontypeable Haemophilus influenzae recognizes sialylated glycoprotein receptors on cultured human epithelial cells; St Geme JW 3rd; Disease due to nontypeable Haemophilus influenzae begins with colonization of the upper respiratory tract mucosa . We recently reported that two surface-exposed high-molecular-weight proteins (HMW1 and HMW2) expressed by a prototypic strain of nontypeable H . influenzae mediate attachment to cultured epithelial cells . In the present study, we examined the nature of the epithelial cell receptor with which HMW1 interacts . Both proteinase K pretreatment and periodate oxidation of epithelial monolayers resulted in a marked decrease in HMW1-mediated binding, suggesting interaction with a glycoprotein structure . Treatment with peptide-N-glycosidase F produced a similar decrease in attachment and thereby provided further evidence for this conclusion . Desialylation of the epithelial cell surface also reduced binding, implying the presence of sialic acid in the receptor structure . Furthermore, lectins specific for terminal alpha 2-3-linked sialic acid were capable of inhibiting HMW1-mediated attachment . In summary, our results indicate that the HMW1 adhesin interacts with a glycoprotein receptor containing N-linked oligosaccharide chains with sialic acid in an alpha 2-3 configuration. Infect Immun, 1994 Sep, 62(9), 3873 - 80 Variable region sequences of a protective human monoclonal antibody specific for the Haemophilus influenzae type b capsular polysaccharide; Lucas AH et al.; A hybridoma secreting a human immunoglobulin G2 kappa monoclonal antibody (MAb) specific for the capsular polysaccharide of Haemophilus influenzae type b (Hib) was isolated . This MAb, designated CA4, was bactericidal to Hib in vitro and protected infant rats from Hib bacteremia . Nucleotide sequence analysis of CA4 variable (V) region cDNA showed that the heavy (H)-chain V region was of subgroup III and was 96% identical to the VH germ line gene segment DP77 (V3-21) . The light (L)-chain V region was of the kappa subgroup III and was 94% identical to the A27 (Humkv325) germ line gene, which is commonly used by rheumatoid factors and other autoantibodies . MAb CA4 did not have rheumatoid factor activity and did not react with histones, DNA, or chromatin . These findings identify an additional VHIII gene segment which can contribute to the anti-Hib capsular polysaccharide repertoire and demonstrate that a VL gene commonly encoding autoantibodies can be utilized for protective immunity. Infect Immun, 1994 Sep, 62(9), 3739 - 44 Platelet-activating factor augments meningeal inflammation elicited by Haemophilus influenzae lipooligosaccharide in an animal model of meningitis; Townsend GC et al.; Research into the pathophysiology of bacterial meningitis has suggested a role for various endogenous inflammatory mediators, such as platelet-activating factor (PAF) . In the present study, rats were inoculated intracisternally with various doses of PAF, with Haemophilus influenzae lipooligosaccharide (LOS) in high doses (20 ng) alone, and with a low dose of LOS (200 pg) with or without low doses of PAF (25 ng to 2.5 micrograms) . Values for cerebrospinal fluid leukocytosis and percent blood-brain barrier permeability to systemically administered 125I-labeled albumin observed after inoculation of low-dose LOS with PAF were greater (P < 0.05) than those observed after inoculation of low-dose LOS alone and not statistically different from those observed after inoculation of high-dose LOS . PAF alone elicited an inflammatory response only at high doses (25 micrograms) . These results support the hypothesis that low cerebrospinal fluid PAF concentrations, such as those observed in children with bacterial meningitis, may augment the inflammatory response to the presence of bacteria in the subarachnoid space. Can Vet J, 1994 Sep, 35(9), 573 - 80 The occurrence of Haemophilus somnus in feedlot calves and its control by postarrival prophylactic mass medication; Van Donkersgoed J et al.; Three field trials were conducted in a large commercial feedlot in Saskatchewan to determine the prevalence of Haemophilus somnus in calves and to evaluate the efficacy of prophylactic mass medication with long-acting oxytetracycline on day 17 (1990, n = 1336), day 11 (1991, n = 4372), or day 8 (1992, n = 5632) postarrival . Hemophilosis accounted for > 40% of the mortality in feedlot calves each year . Haemophilus somnus was cultured from the blood of one febrile calf on day 1 (0.1%, n = 895), but it was not cultured from nasal swabs on day 1 or day 11 (n = 881) or from blood samples on day 11 (n = 883) . Similarly, it was not cultured from nasal swabs or blood samples from sick calves first treated for bovine respiratory disease (BRD) (n = 219) . Serological titers to H . somnus increased (p < 0.05) in unvaccinated calves from day 1 (Geometric mean titer = 11,846) to day 96 (Geometric mean titer = 63,712), indicating natural infection following feedlot entry . Calves that relapsed twice with BRD or died from BRD +/- hemophilosis had significantly (p < 0.06) lower titers to H . somnus on days 1 and 96 than those that did not relapse twice or die . Postarrival mass medication with long-acting oxytetracycline did not reduce (p > 0.05) the risk of hemophilosis mortality . However, it reduced (p < 0.05) the risk of BRD treatment by 14% and the risk of BRD mortality by 71% . Additional epidemiological studies of H . somnus are needed so that we can develop strategic medication and vaccination programs to reduce losses from hemophilosis. Jpn J Antibiot, 1994 Sep, 47(9), 1219 - 30 {A clinicobacteriologic study on sultamicillin fine granules in pediatric sinusitis}; Sugita R et al.; We carried out clinical and bacteriological studies on sultamicillin (SBTPC) in pediatric sinusitis at 10 general practice settings . The results are summarized as follows . 1 . The major isolated organisms from purulent nasal discharges were Streptococcus pneumoniae 27.5%, Haemophilus influenzae 32.4% and Moraxella catarrhalis 9.9% . Similar results were observed for the major isolates from nasopharynx . 2 . 33% of the isolated S . pneumoniae were penicillin-insensitive S . pneumoniae (PISP) against which the MICs were equal to or higher than 0.1 microgram/ml . 3 . PISP was isolated from 14% of all cases . 4 . The clinical efficacy rate was 77.5% and was deemed satisfactory . 5 . In the bacteriological study, persistence rate of PISP was 38.5% among the PISP from purulent nasal discharge and 60.0% among the PISP from nasopharynx which and these values were significantly higher than persistence rates of PSSP, H . influenzae and M . catarrhalis . 6 . Adverse reactions were observed in 21.5% of all cases, involving diarrhea and loose stool. Jpn J Antibiot, 1994 Sep, 47(9), 1202 - 9 {The clinical study of cefpodoxime proxetil dry syrup preparation in the pediatric field}; Kasagi T et al.; The clinical efficacy was examined for the newly developed oral cephem antibiotic, cefpodoxime proxetil (CPDX-PR) dry syrup, in the treatment of various acute infections in the field of pediatrics . CPDX-PR dry syrup was administered at 10 mg/kg/day in 3-divided doses to 535 children at 21 institutions, including Tottori University Hospital and its related hospitals . The efficacy rate of this drug was determined to be 80.8% . Among isolates, Staphylococcus aureus and Streptococcus sp . were highly susceptible to the drug, whereas Haemophilus influenzae showed relatively poor susceptibility . Side effects were observed in 2.80% of all of the patients, and abnormal laboratory findings were detected in 1.87% . The low incident of side effects demonstrated its high safety, and this drug was considered to be very useful for such pediatric infections as acute tonsillitis, acute pharyngitis and acute bronchitis. Jpn J Antibiot, 1994 Sep, 47(9), 1186 - 91 {Antimicrobial activities of fosfomycin against Streptococcus pneumoniae and Haemophilus influenzae recently observed in sinusitis patient}; Deguchi K et al.; In order to examine antimicrobial activities of fosfomycin (FOM), the minimum inhibitory concentrations (MICs) of FOM and those of control drugs were determined against Streptococcus pneumoniae and Haemophilus influenzae isolated from sinusitis patients from September to November, 1993, and the following results were obtained . 1 . Among 50 S . pneumoniae strains tested, there were 10 strains (20.0%) of benzylpenicillin (PCG)-insensitive S . pneumoniae (PISP) and 2 strains (4.0%) of PCG-resistant S . pneumoniae (PRSP); but the MIC distributions of FOM among the PISPs and the PRSPs were almost identical to those among the PCG-susceptible S . pneumoniae (PSSP) . 2 . There were 12 strains (24.0%) of beta-lactamase producing strains among 50 strains of H . influenzae tested, but the FOM's MIC distribution among these strains was almost identical to that among beta-lactamase non-producing strains . 3 . The results obtained on the MIC90s of FOM against S . pneumoniae and H . influenzae suggest that the nebulization treatment with FOM nasal preparation satisfies the condition "above the MIC". J Clin Pathol, 1994 Sep, 47(9), 796 - 8 Rapid organism identification from Bactec NR blood culture media in a diagnostic microbiology laboratory; Claxton PM et al.; AIMS--To evaluate rapid organism identification on positive blood culture Bactec NR media (phial types 26, 27, 42 and 17), and to assess the usefulness of these procedures in a diagnostic microbiology laboratory . METHODS--Two hundred and sixty, first positive, blood culture bottles from individual patients were tested by rapid identification methods selected on the basis of Gram film organism morphology . Tube coagulase and latex agglutination were applied to presumptive staphylococci; latex agglutination antigen detection methods to suspected pneumococci, Neisseria and Haemophilus sp; and latex agglutination grouping tests for cultures thought to be non-pneumococcal streptococci . RESULTS--Media type did not influence test performance (p > 0.05 for all comparisons) . Misapplication of methods occurred on eight occasions and there were 14 false positive results, nine involving the latex reagents for group C streptococci and pneumococci . The positive predictive values for tube coagulase tests and latex reactions for H influenzae type b, and N meningitidis groups B and C were 100% . The pneumococcal and staphylococcal latex tests gave positive predictive values of 94.1% and 62.5%, respectively, and the corresponding figure for streptococcal grouping reactions was 75.9% . With the exception of staphylococcal latex testing (80%) all investigation negative predictive values were > 90% . CONCLUSIONS--The performance of the staphylococcal latex agglutination method was unsatisfactory and it is not appropriate for use with the media studied . In view of the cross-reactions observed with the tests used to identify group C streptococci and pneumococci, positive findings must be interpreted with caution . In all other regards the protocol evaluated produced rapid, reliable, clinically useful information and, subject to local experience, is recommended to users of Bactec NR media. Chemotherapy, 1994 Sep-Oct, 40(5), 299 - 303 Antibiotic resistance patterns of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis: a prospective study in Murcia, Spain, 1983-1992; Gomez J et al.; We prospectively determined the role of commonly used antibiotics in the emergence of antimicrobial resistance among the predominant pathogens associated with the respiratory tract . Clinical isolates of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis from otic exudates, exudates obtained by puncture of the paranasal sinuses, sputum samples or blood cultures obtained from hospital inpatients with symptoms of significant upper or lower respiratory tract infection were analyzed . Between 1983 and 1992 a statistically significant increase in strains of S . pneumoniae and H . influenzae resistant to ampicillin, erythromycin and co-trimoxazole was detected . A significant increase in strains of M . catarrhalis resistant to ampicillin was also found . The increase in the consumption of aminopenicillins, co-trimoxazole and macrolides was associated with an increase in strains of S . pneumoniae, H . influenzae and M . catarrhalis resistant to these agents. Plasmid, 1994 Sep, 32(2), 228 - 32 An analysis of the complete nucleotide sequence of the Haemophilus ducreyi broad-host-range plasmid pLS88; Dixon LG et al.; We present an analysis of the complete nucleotide sequence of pLS88, a naturally occurring, 4.8-kb broad-host-range plasmid isolated from Haemophilus ducreyi and encoding resistance to sulfonamides, streptomycin, and kanamycin . Sequence analysis of the genes encoding sulfonamide and streptomycin resistance revealed homology to the RSF1010 sulII and strA genes . The sulII-strA intergenic region of pLS88 has a 38-bp deletion identical to that of the RSF1010-like plasmid pHD8.1, isolated from Actinobacillus pleuropneumoniae . The kanamycin resistance gene shows strong homology to Tn903, but lacks the inverted repeats of the transposon . No other genes have been identified . The region downstream of the kanamycin resistance gene shows homology to the RSF1010 oriV region; however, this region is not essential to plasmid replication . The ori of pLS88 is contained within a 1060-bp region and does not appear to contain structures typical of plasmid origins . This region is flanked by DNA showing strong homology to regions both upstream and downstream of the Haemophilus influenzae ROB-1 beta-lactamase gene . Because of the small size of the origin, pLS88 appears to resemble the structure of narrow-host-range plasmids, but replicates, via an as yet unidentified mechanism, as a broad-host-range plasmid. Cent Afr J Med, 1994 Sep, 40(9), 234 - 44 Teenage obstetric and gynaecological problems in an African city; Duncan ME et al.; OBJECTIVE: To measure the prevalence of sexually transmitted diseases (STD), pelvic inflammatory disease (PID), cervical cancer, pregnancy and use of contraception in teenagers, and to determine socioeconomic factors associated with these conditions to aid planners of medical services and promotion of sexual health . SUBJECTS: 181 Ethiopian teenagers and 1,845 women aged 20 to 45 years for comparison . SETTING: Gynaecological outpatient department, antenatal, postnatal and family planning clinics, in two teaching hospitals and a mother and child heath centre in Addis Ababa, Ethiopia . METHODS: Results of serologic tests for STD, clinical evidence of PID, and cervical cytology were analysed against socio-economic factors . RESULTS: In teenagers early age at first marriage/coitus, more common in those of rural origin, was associated with poverty, a greater number of lifetime sexual partners, and prostitution: 40 pc were first sexually active before the menarche . Prevalence of seropositivity to specific STD pathogens was; Treponema pallidum (TPHA) 21 pc, Neisseria gonorrhoeae (gonococcal antibody test: GAT) 40 pc, genital chlamydiae 51 pc, hepatitis B virus 36 pc, herpes simplex virus (HSV-2) 32 pc, and Haemophilus ducreyi 16 pc: 92 pc of teenagers were seropositive to one or more STD's . STD seroprevalence was higher in those with more than one sexual partner, those sexually active by age 15 (very high in those sexually active by age 12), those involved in prostitution and those attending the family planning clinic . Forty three pc had clinical evidence of PID; one married at age 10 had invasive cervical cancer by age 18; 40 pc of teenagers were pregnant compared with 25 pc of those aged 20 to 45; 21 pc attended for family planning; of regular FPC attenders 81 pc were GAT seropositive . CONCLUSION: Despite legislation early age of sexual debut is common, STD and PID are widely prevalent, the pregnancy rate in adolescents is high and contributes to the national population growth rate . Action is required at family, medical and governmental level to encourage cultural acceptance that marriage and sexual activity should not occur before the age of 16 years, with education appropriate to culture to prevent STD . Similar studies are recommended in other countries to establish a baseline for informed strategy regarding prevention of STD and health educationPIP: A survey of 181 Ethiopian females ages 14-19 years recruited from health facilities in Addis Ababa revealed a high incidence of obstetric and gynecologic problems . All subjects completed a questionnaire administered by a female health worker and underwent a gynecologic examination and serologic tests . 49% of subjects were married and 18% were divorced; 11% were prostitutes . Age at first intercourse was under 12 years in 18%, 13-15 years in 38%, and 16 years or above in 44%; 40% were sexually active before menarche . 92% of adolescents had at least one sexually transmitted disease (STD), predominantly gonorrhea (40%), genital chlamydia (51%), hepatitis B (36%), herpes simplex virus (32%), and syphilis (21%), and 43% had clinical signs of pelvic inflammatory disease (PID) . 53% had had at least one pregnancy . The earlier the age at first intercourse, the more likely it was that the adolescent would have multiple sexual partners and several STDs; adolescents in this category were also more likely to be from poor families from rural areas . Only 21% were attending a family planning clinic for annual check-ups; 14% of these females were using contraception . Although only 8% were infertile at the time of assessment, 23% had clinical evidence of salpingitis--a risk factor for future infertility . Given the long-term health risks (e.g., infertility, cervical cancer, and gonorrhea-related infant morbidity) associated with the patterns observed among these adolescents, it is recommended that STD education receive higher priority and that the Ethiopian Government consider greater enforcement of the law prohibiting sexual intercourse and marriage before the age of 16 years . Semin Respir Infect, 1994 Sep, 9(3), 180 - 8 Community-acquired pneumonia: the future of the microbiology laboratory: focused diagnosis or syndromic management? MacDonald KS, Scriver SR, Skulnick M, Low DE. The traditional classification of community-acquired pneumonia into typical and atypical pneumonia to facilitate successful empirical treatment is no longer optimal . An accurate prediction of cause and adequate empirical therapy cannot be provided with this approach in severely ill patients . There is an increasing spectrum of recognized treatable pathogens presenting as community-acquired pneumonia including Legionella species, Chlamydia pneumoniae, and Pneumocystis carinii in addition to the traditional community pathogens . The variability of presentation in severely ill or compromised hosts makes clinical prediction of cause inadequate . A more rational approach may involve the classification of patients by the severity of illness and underlying disease with little or no microbiological workup in mild illness unless the results will contribute to the epidemiological surveillance of resistance because these investigations have not been shown to affect outcome in this setting . Etiologic diagnosis should be more aggressively sought and the microbiology laboratory can be best used by providing the efficient and rapid diagnosis of this expanded range of pathogens in more severely ill patients . The mounting antimicrobial resistance of common pathogens such as Streptococcus pneumoniae, Haemophilus influenzae, and Staphylococcus aureus will require not only a critical review of empirical therapy, but an increased emphasis on epidemiological monitoring of resistance by laboratories and effective communication with clinicians. Semin Respir Infect, 1994 Sep, 9(3), 166 - 79 Initial investigation and treatment of the patient with severe community-acquired pneumonia; Ortqvist A; Approximately 5% to 10% of patients hospitalized with community-acquired pneumonia (CAP) require treatment in an intensive care unit (ICU) because of severe disease . The case fatality rates in these patients are high, between 20% and 50% in most series . Streptococcus pneumoniae, Legionella spp, Haemophilus influenzae, Staphylococcus aureus, and gram-negative enteric bacteria are the most common causes of severe CAP . However, because the spectrum of pathogens encountered in these patients is unlimited, including viruses, tuberculosis, and opportunistic pathogens, it is crucial to obtain an etiologic diagnosis . A complete diagnostic arsenal should be used, including, if possible, invasive diagnostic procedures before antibiotics are given . The initial empirical therapy must cover the most common pathogens, and in most patients this can be accomplished with a combination of a cephalosporin and a macrolide. Blood Rev, 1994 Sep, 8(3), 179 - 91 Prophylaxis against late infection following splenectomy and bone marrow transplant; Fielding AK; There is a well documented risk of late infection following both splenectomy and bone marrow transplantation . In asplenic patients, the phagocytic and antibody producing roles of the spleen are lost and there is a lifelong susceptibility to infection which may be overwhelming and fatal . Patients most at risk are children, those with underlying lymphoproliferative disorders and those receiving immunosuppressive therapy . Although it is hard to prove benefit from preventative strategies, patients are likely to benefit from prophylactic antibiotic therapy and from immunisation with pneumococcal, Haemophilus influenzae-B and meningococcal vaccine given prior to splenectomy . Following an allogeneic bone marrow transplant (BMT), recovery of immune function takes up to a year . During this time, patients are at high risk from cytomegalovirus (CMV) and varicella zoster virus (VZV) infections and also from pneumocystis pneumonia . Prophylactic medications are used to good effect . The major threat of late infection occurs in patients with chronic graft versus host disease (cGVHD)--there is increased susceptibility to bacterial, fungal and viral infections . Many patients without cGVHD recover immune function fully and many develop antibodies to specific recall antigens . This does not occur in all patients and although there is a low risk of infection with organisms against which vaccines are available . If it is not possible to measure specific antibody titres and consequently offer selective re-immunisation, then a universal vaccination strategy should be in force . Response to vaccines is likely to be poor before one year post BMT . For autologous transplant recipients, immune recovery is probably complete and routine re-immunisation is not likely to offer much benefit . For both asplenic and bone marrow transplant patients, education of patient and physician is important. Int J STD AIDS, 1994 Sep-Oct, 5(5), 332 - 7 Seroprevalence and incidence of sexually transmitted diseases in a rural Ugandan population; Wagner HU et al.; The aim of the study was to determine in a rural population the age- and sex-specific prevalence and incidence rates of serological reactivity of 5 common sexually transmitted diseases (STDs) and their association with HIV-1 antibody status . Of the adult population of two villages (529 adults aged 15 years or more) 294 provided an adequate blood specimen both on enrollment and at 12 months . The sera were tested at 3 collaborating laboratories for antibodies against HIV-1, Treponema pallidum, Haemophilus ducreyi, Chlamydia trachomatis and herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) . A sample of 45 children were tested for HSV-1 and HSV-2 . Seroprevalence rates in adults on enrollment were 7.8% for HIV-1, 10.8% for active syphilis, 10.4% for H . ducreyi, 66.0% for C . trachomatis, 91.2% for HSV-1 and 67.9% for HSV-2 . Males were significantly more likely than females to be seropositive for H . ducreyi (15.6% versus 6.6%), but less likely to be HSV-2 antibody positive (57.0% versus 74.4%) . Reactivity to H . ducreyi, C . trachomatis and HSV-2 rose with increasing age . In contrast, active syphilis showed no age trend . All STDs tended to be more common in those HIV-1 seropositive . Incidence rates over the 12 months were nil for HIV-1, 0.5% for syphilis, 1.2% for H . ducreyi, 11.3% for C . trachomatis, and 16.7% for HSV-2 . The results of this exploratory study indicate that all STDs included are common in this rural population . The high HSV-2 prevalence rate among adolescents suggests that HSV-2 may be an important risk factor for HIV-1 infection.(ABSTRACT TRUNCATED AT 250 WORDS) PIP: A seroprevalence survey conducted in rural Uganda revealed a high potential for interaction between sexually transmitted diseases (STDs) such as herpes simplex virus type 2 (HSV-2) and human immunodeficiency virus (HIV) . Venous blood samples were collected at baseline and one year later from 294 randomly selected adults aged 15 years or over from two neighboring villages . At baseline, 23 (7.8%) adults were HIV-positive; no seroconversion occurred during the one-year study period . STD prevalence rates were 10.8% for syphilis, 10.4% for Hemophilus ducreyi, 66.0% for Chlamydia trachomatis, and 91.2% for HSV-1 and 67.9% for HSV-2 . More females (74.4%) than males (57.0%) were HSV-2 antibody-positive . Reactivity to H . ducreyi, C . trachomatis, and HSV-2 rose with increasing age, but there was no such trend for syphilis . HIV prevalence rates were 0.0% among those with no serologic evidence of previous STDs, 2.6% among those with one or two prior STDs, and 20.0% among those with three or four STD markers . Of particular concern was the high rate of HSV-2 prevalence among adolescents (85% among females aged 20-24 years and 82% in males aged 25-29 years) . It is suggested that age-specific HSV-2 seroprevalence can provide an accurate marker of premarital sexual activity among Ugandan adolescents since it lacks the potential for bias associated with self-reporting in this population . Sex Transm Dis, 1994 Sep-Oct, 21(5), 280 - 8 Seroepidemiological studies of Haemophilus ducreyi infection in Ethiopian women; Duncan ME et al.; BACKGROUND AND OBJECTIVES: To measure prevalence of anti-Haemophilus ducreyi antibodies in sera from Ethiopian female attendees, and to determine significant socioeconomic associations . STUDY DESIGN: A modified ELISA immunoassay was used to test sera of 1,831 Ethiopian women attending gynecological, obstetric, and family planning clinics in Addis Ababa . RESULTS: Overall seropositivity was 19.4% . Prevalence rates for seropositivity for antibodies to H . ducreyi were significantly associated with ethnic group and religion, older age (> or = 50 years: 28%), early age at first coitus (< 13 years: 28%) and first coitus before the menarche (25%), being divorced (27%) or a prostitute (24%), longer duration of marriage (> 20 years: 27%) and sexual life (> 20 years: 24%), number of lifetime sexual partners (2 to 5 partners: 27%) and self-reported history of both syphilis and gonorrhea (31%) . Of these factors, the two most significant were first coitus before the menarche (P < 0.0001) and not being still married to the first husband/sexual partner (P < 0.001) . Differences in seropositivity according to ethnic group and religion may be explained by the number of women within each group who had only one lifetime sexual partner . Women with serological evidence of exposure to another sexually transmitted disease (STD) had a greater risk of exposure to H . ducreyi . The odds ratio for H . ducreyi seropositivity in women with syphilis or gonorrhea was 3.6, for women with genital chlamydial infection, 2.3, and for those with HBV or HSV-2, 1.4 and 1.3 respectively . CONCLUSIONS: This study illustrates the usefulness of the modified ELISA immunoassay for measuring exposure to H . ducreyi, and the usefulness of H . ducreyi as a marker for cumulative sexual exposure . Further studies on the association of HIV transmission and H . ducreyi in Ethiopia are now indicatedPIP: Genital ulcerated disease (GUD), which includes chancroid, has been identified as a risk factor for HIV transmission . This study reports the prevalence of anti-Hemophilus ducreyi (chancroid) antibodies in 1831 Ethiopian women and looks at the behavioral and social factors which might affect the incidence and potential spread of chancroid . Patient data regarding ethnic and socioeconomic aspects were collected from detailed questionnaires . Blood collection was performed under medical surveillance . Complete gynecological examinations were performed . Papanicolaou stained smears were used as the basis of the cytological data . Serological studies utilized an enzyme immunoassay (EIA) test for STD detection . Statistical tests used included the Chi-square test, the multivariate analysis technique, and the Cochran-Mantel-Haenszel General Association Statistic Test . Antibodies to H . ducreyi were found in 335 women (19.4%) . Prevalence of H . ducreyi was significantly associated with Amhara or Tigre ethnic heritage; older age; first coitus before beginning menstruation; history of STDs; divorced status; being a prostitute; longer duration of married and sexual life; and younger age at first coitus . Logistic regression demonstrated that 3 factors were significant when associated with H . ducreyi seropositivity . First coitus before beginning menstruation was highly significant (OR 1.95; 95% CI, 1.49-2.57; P 0.0001) . Not being still married to the first husband was also significant (OR 1.68; 95% CI, 1.23-2.30; P 0.001) . Being of the Ethiopian Orthodox religion was significant (OR 2.11; 95% CI, 1.21-3.68; P 0.005) . Prevalence in women with 2-5 lifetime husbands was higher than in women with only 1 husband . Sex Transm Dis, 1994 Sep-Oct, 21(5), 247 - 57 Characterization of the cytopathic effect of Haemophilus ducreyi; Hollyer TT et al.; BACKGROUND AND OBJECTIVES: Haemophilus ducreyi is the etiologic agent of chancroid, which is a genital ulcer disease that increases the risk of acquiring and transmitting HIV . The pathogenesis of H . ducreyi is not well understood . GOAL OF THIS STUDY: The goal of this study was to use a quantitative tetrazolium-based XTT assay to characterize the cytopathic effect of H . ducreyi on human foreskin fibroblasts . STUDY DESIGN: Haemophilus ducreyi strains 35000, R018, A77 and CIP542 were evaluated using the XTT assay . The role of attachment on resultant CPE was assessed using a wash step 2 hours post-infection . Internalization was evaluated by the gentamicin kill assay . Secreted exotoxin was studied using permeable inserts to separate the bacteria from the HFF monolayer . RESULTS: HFF cell damage did not appear to be mediated by a secreted H . ducreyi cytotoxin . Direct contact of viable H . ducreyi with HFF cells was required for cell damage . H . ducreyi strains that attached poorly could be readily removed by a wash step . This reduced their capacity to damage HFF cells significantly . Although some H . ducreyi strains attach to high levels within 4 hours, no HFF cell damage was detected by the XTT assay . However, once HFF cell damage was detected by 24 hours, it was not easily reversible, despite antibiotic treatment that eradicated H . ducreyi . Internalization of H . ducreyi by HFF cells apparently did not occur to a significant degree . CONCLUSION: This study indicates that classic "soluble exotoxins" are not likely the key component in H . ducreyi pathogenesis . Attachment or direct contact with HFF cells are required for H . ducreyi to cause a CPE. Mol Microbiol, 1994 Sep, 13(5), 863 - 73 The 100 kDa haem:haemopexin-binding protein of Haemophilus influenzae: structure and localization; Cope LD et al.; All Haemophilus influenzae strains have an absolute requirement for exogenously supplied haem for aerobic growth . A majority of strains of H . influenzae type b (Hib) produce a 100 kDa protein which binds haem: haemopexin complexes . This 100 kDa haem:haemopexin binding protein, designated HxuA, was originally detected on the Hib cell surface . Monoclonal antibody (mAb)-based analyses revealed that the HxuA protein was also present in soluble form in Hib culture supernatants . This soluble HxuA protein exhibited haem:haemopexin-binding activity in a direct binding assay . Nucleotide sequence analysis of the hxuA gene from Hib strain DL42, together with N-terminal amino acid analysis of HxuA protein purified from Hib culture supernatant, revealed that this protein was synthesized as a 101 kDa precursor with a leader peptide that was removed to yield a 99 kDa protein . Southern blot analysis of chromosomal DNA from four Hib and four non-typeable H . influenzae (NTHI) strains detected the presence of a single band in each strain that hybridized a Hib hxuA gene probe . Subsequent analysis of these NTHI strains showed that all four strains released into culture supernatant a haem:haemopexin-binding protein that migrated in SDS-PAGE at a rate similar or identical to that of the Hib HxuA protein . A Hib hxuA mutant was used to screen an NTHI genomic DNA library and an NTHI gene was cloned that complemented the mutation in this Hib strain . Nucleotide sequence analysis of this NTHI gene revealed that it encoded a protein with 87% identity to the Hib HxuA protein.(ABSTRACT TRUNCATED AT 250 WORDS) Lung Cancer, 1994 Sep, 11(3-4), 243 - 9 Pulmonary infections in lung cancer patients at diagnosis; Putinati S et al.; We carried out a prospective study involving 96 consecutive lung cancer patients at diagnosis, in order to determine through quantitative cultures of the bronchoalveolar lavage (BAL) fluid, the prevalence of pulmonary infections; we also evaluated the relationship between a patient's performance status, immunocompetence, lung cancer stage, histotype and the occurrence of respiratory infections . The patients (81 males, 15 females) had a mean age of 64 +/- 9 years . Of these, 62 were smokers, 30 were ex-smokers and four had never smoked . Sixty-seven patients had a prior history of chronic bronchitis . A total of 42 micro-organisms were cultured from the BAL fluids of 33 patients (34.3%) . Fifty percent of these micro-organisms were gram-negative, 33.3% were gram-positive and the remaining 16.7% were other micro-organisms . The bacilli most often isolated were the Haemophilus species, accounting for 38.8% of all gram-negative bacilli . The most frequently isolated gram-positive pathogen was the Staphylococcus aureus . We have not found a significant relationship between the presence of a respiratory infection and the different cell types separately analyzed, nor with SCLC and NSCLC patient groups, nor with the stage of the disease . The performance status, the immunoregulatory ratio and the lymphocyte subsets were not significantly different in patients with or without a pulmonary infection . We think that the identification of a definite etiologic agent is of great importance for a rational anti-microbial treatment of pulmonary infections. Antimicrob Agents Chemother, 1994 Sep, 38(9), 2003 - 7 Evaluation of antimicrobial activities of clarithromycin and 14-hydroxyclarithromycin against three strains of Haemophilus influenzae by using an in vitro pharmacodynamic model; Walker KJ et al.; An in vitro pharmacodynamic model was used to simulate the in vivo pharmacokinetics of clarithromycin and 14-hydroxyclarithromycin in order to generate time-kill curves for three clinical isolates of Haemophilus influenzae (isolates 2019, 91-183, and 1746) . Representative concentrations in serum or lung tissue and the pharmacokinetic parameters of clarithromycin and the 14-hydroxy metabolite, separately and in combination, were simulated for the time-kill studies . Amoxicillin-clavulanic acid was used as a control drug . The simulation of typical concentrations of the macrolides in serum in time-kill studies resulted in magnitudes of bacterial killing that were less than (for strains 2019 and 91-183, MICs = 4 mg/liter for clarithromycin and 14-hydroxy-clarithromycin) or equal to (for strain 1746, MIC = 1 mg/liter for clarithromycin and 14-hydroxyclarithromycin) those observed in amoxicillin-clavulanic acid studies . When typical concentrations in lung tissue were simulated, total log decreases in bacterial counts were greater than those achieved with typical concentrations in serum and, in the case of strain 1746, exceeded the magnitude observed with the control drug . In each case, the time to 3-log-unit killing was longer for the macrolides than for amoxicillin-clavulanic acid . Time-kill curve analyses demonstrated the presence of synergy (defined as a 2-log-unit decrease in the CFU per milliliter between the combination and the most active constituent at any time point) for the combination of clarithromycin and 14-hydroxyclarithromycin at simulated concentrations in serum for one strain of H . influenzae (isolate 91-183) . Synergism is likely bacterial strain specific, and the presence of synergy may be dependent on the antibiotic concentrations that are tested . Evaluation of the kill curve kinetics in terms of bactericidal rate for the various starting concentrations of clarithromycin did not result in a clear demonstration of either concentration-dependent or concentration-independent bactericidal activity. Vet Immunol Immunopathol, 1994 Sep, 42(3-4), 349 - 56 Compartmentalization of specific B-cells in sheep mucosae associated lymphoid organs; Zanin C et al.; Numerous studies have shown that Peyer's patches (PP) contribute to the seeding of other lymphoid organs in sheep . This was demonstrated by perfusing labeled lymphocytes in PP, and later investigating their presence in drainage lymph nodes, spleen, peripheral blood or bone marrow . These data showed that PP export considerable numbers of cells every day, but provided no information as to their specificity . In this work, we used the enzyme-linked immunosorbent assay (ELISA) spot method to investigate, in the peripheral blood, mesenteric and cervical lymph nodes and tonsils from ten sheep, the numbers of specific B-cells, directed to four common bacteria of the oro-pharyngeal area of mammals: Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae or Klebsiella pneumoniae . The data were obtained from five sets of monozygous sheep, one animal of each pair being previously fed ribosomal preparations of these bacteria . Both prior to and after oral challenge, specific B-cells could be found in all the tissues tested . They were mostly IgG-producing cells and preferentially located in oro-pharyngeal drainage lymph nodes and tonsils . Their numbers increased in these lymph nodes after stimulation, while they decreased in mesenteric lymph nodes . These observations are consistent with the current hypothesis suggesting intestinal sensitization, proliferation and fast emigration of specific B-cells after oral challenge. J Infect, 1994 Sep, 29(2), 203 - 5 Neonatal osteomyelitis in Down's syndrome due to non-encapsulated Haemophilus influenzae; Williams R et al.; A case of neonatal osteomyelitis in a baby with Down's syndrome is described . The causative organism was a non-encapsulated Haemophilus influenzae biotype I, which was isolated from pus at the site of infection . This organism has not previously been reported as a cause of neonatal osteomyelitis. Microb Pathog, 1994 Sep, 17(3), 167 - 74 A quantitative chemiluminescent ribosomal probe method for monitoring adherence of Haemophilus ducreyi to eukaryotic cells; Alfa MJ et al.; This study evaluated a commercially available chemiluminescent-labelled, ribosomal RNA-directed DNA probe (CRP) as a method to quantitate attachment of H . ducreyi to human foreskin fibroblast (HFF) cells . Evaluation of four strains of H . ducreyi demonstrated that the CRP assay was unaffected by eukaryotic cells and its advantages were: (i) quantitation was done after attachment so it did not interfere with the attachment process, and (ii) it was a rapid, reliable method for quantitating bound bacteria, despite bacterial clumping . Gentamicin-killed H . ducreyi attached to both HFF cells and viable controls, suggesting that the adhesins are components constitutively present on the surface of H . ducreyi . This method may be widely applicable, since the probe recognizes most prokaryotic rRNA sequences. J Bacteriol, 1994 Sep, 176(18), 5835 - 42 Sequencing analysis reveals a unique gene organization in the gyrB region of Mycoplasma hominis; Ladefoged SA et al.; The homolog of the gyrB gene, which has been reported to be present in the vicinity of the initiation site of replication in bacteria, was mapped on the Mycoplasma hominis genome, and the region was subsequently sequenced . Five open reading frames were identified flanking the gyrB gene, one of which showed similarity to that which encodes the LicA protein of Haemophilus influenzae . The organization of the genes in the region showed no resemblance to that in the corresponding regions of other bacteria sequenced so far . The gyrA gene was mapped 35 kb downstream from the gyrB gene. Infect Immun, 1994 Sep, 62(9), 3712 - 22 Mapping of bactericidal epitopes on the P2 porin protein of nontypeable Haemophilus influenzae; Haase EM et al.; The P2 porin protein is the major outer membrane protein of nontypeable Haemophilus influenzae and is a potential target of a protective immune response . Nine monoclonal antibodies (MAbs) to P2 were developed by immunizing mice with nontypeable H . influenzae whole organisms . Each MAb reacted exclusively with the homologous strain in a whole-cell immunodot assay demonstrating exquisite strain specificity . All nine MAbs recognized abundantly expressed surface-exposed epitopes on the intact bacterium by immunofluorescence and immunoelectron microscopy . Each MAb was bactericidal to the homologous strain in an in vitro complement-mediated killing assay . Immunoblot assay of cyanogen bromide cleavage products of purified P2 indicated that MAb 5F2 recognized the 10-kDa fragment, and the other eight MAbs recognized the 32-kDa fragment . Competitive ELISAs confirmed that 5F2 recognized an epitope that is different from the other eight MAbs . To further localize epitopes, MAbs 5F2 and 6G3 were studied in protein footprinting by using reversed-phase high-performance liquid chromatography . Three potential epitope-containing peptides which were reactive in an enzyme-linked immunosorbent assay with both 5F2 and 6G3 were isolated . These peptides were identified by N-terminal amino acid sequence and localized to loops 5 and 8 of the proposed model for P2 . Fusion proteins consisting of glutathione S-transferase fused with variable-length peptides from loops 5 and 8 were expressed in the pGEX-2T vector . Immunoblot assay of fusion peptides of loops 5 and 8 confirmed that 5F2 recognized an epitope within residues 338 to 354 of loop 8; 6G3 and the remaining MAbs recognized an epitope within residues 213 to 229 of loop 5 . These studies indicate that nontypeable H . influenzae contains bactericidal epitopes which have been mapped to two different surface-exposed loops of the P2 molecule . These potentially protective epitopes are strain specific and abundantly expressed on the surface of the intact bacterium. J Biol Chem, 1994 Aug 19, 269(33), 21340 - 5 Binding sites for bacteriophage HP1 integrase on its DNA substrates; Hakimi JM et al.; The temperate phage HP1 integrates its genome into the chromosome of Haemophilus influenzae by site-specific recombination between host and phage DNA segments, the attachment sites . This reaction is promoted by the HP1-encoded integrase . The interactions of HP1 integrase with its DNA substrates have been characterized by DNase I footprinting . Two classes of binding sites were identified . At sites of type I, integrase binding almost completely eliminated cleavage by DNase I; type I sites shared the consensus sequence 5'-AGGGATTTWW . At type II sites, integrase binding produced alternating regions of protection from and enhancement of cleavage, suggesting that binding at these sites distorted the DNA . The consensus sequence for type II sites was 5'-ACTGGCGRTW . Each binding site contained two copies of the relevant consensus . The host attachment site (attB) contains an inverted pair of type I consensus sequences surrounding the strand exchange points . The phage attachment site (attP) includes six binding sites, three of type I and three of type II, distributed along its 500 nucleotide pairs . All type I sites contain two consensus motifs arranged as inverted repeats . One of these surrounds the strand exchange points in this substrate, one is located internally, and the third coincides with the right boundary of the attP sequence . One type II site, consisting of an inverted repeat of two type II consensus motifs, coincides with the left boundary of the attP sequence . The other two type II sites contain directly repeated pairs of the consensus and are internally located. Gene, 1994 Aug 19, 146(1), 95 - 100 Sequence of the rec-2 locus of Haemophilus influenzae: homologies to comE-ORF3 of Bacillus subtilis and msbA of Escherichia coli; Clifton SW et al.; The nucleotide sequence of a 4243-bp PstI fragment containing the rec-2 gene of Haemophilus influenzae was determined . The amino acid (aa) sequences of four putative proteins were deduced from the corresponding open reading frames (ORFs) . The 2400-bp ORF2 accounted for rec-2, based on the sequences of DNA fragments that contained rec-2::mini-Tn10 mutations . The rec-2 gene encoded a putative 800-aa protein with a M(r) of 90,561 . Sequence analysis suggested that the rec-2 product contained nine highly probable integral membrane-spanning segments . Database searches showed that rec-2 was homologous to the comE-ORF3 gene of Bacillus subtilis . This hypothesis is consistent with the known involvement of both of these genes in the passage of transforming DNA through the competent-cell envelope . Although the sequences of the other three ORFs were incomplete, sufficient data were available to allow inferences about their homologies to other genes . ORF4, which overlapped ORF1, was homologous to the Escherichia coli dnaK suppressor gene, dksA, and therefore was named dsh-1 (dnaK suppressor homolog) . Mutations in dsh-1 and its putative promoter region caused a mild sensitivity to UV light, but did not affect DNA recombination . ORF3, located downstream from rec-2, was homologous to msbA, an essential gene of E . coli with extensive similarity to the ATP-dependent translocators . ORF3 was named msh-1 (msbA homolog) . Mutations in msh-1 had no effects on genetic transformation . The close juxtaposition of rec-2 and msh-1 implied that the expression of msh-1 could be linked to the translation of the rec-2 ORF. Gene, 1994 Aug 19, 146(1), 101 - 3 Cloning and sequencing of the Haemophilus influenzae ssb gene encoding single-strand DNA-binding protein; Jarosik GP et al.; The ssb gene of Haemophilus influenzae was cloned and sequenced . The deduced protein possessed 61 and 60% identity with the Serratia marcescens and Escherichia coli SSB proteins, respectively . H . influenzae ssb was also shown to complement an E . coli ssb-1 mutation. Lancet, 1994 Aug 6, 344(8919), 362 - 6 Efficacy of Haemophilus influenzae type b conjugate vaccine PRP-T; Booy R et al.; Efficacy of the Haemophilus influenzae type b (Hib) conjugate vaccine PRP-T (Pasteur-Merieux) was evaluated in a controlled community intervention study in the Oxford region, UK . PRP-T was offered to infants from May 1, 1991 in three of the region's eight districts and from July 1, 1991, in a fourth district . It was given by separate injection in addition to the standard diphtheria, tetanus, and pertussis vaccine according to an accelerated 2, 3, and 4 month schedule without a booster dose in the second year of life . By October 1, 1992, more than 90% of infants in vaccine districts had received at least one dose of PRP-T . None of the infants given three doses had developed Hib infection, whereas 11 infections occurred in the control population (vaccine efficacy 100%, 95% CI 80-100%) . Intention-to-treat analysis also showed a high estimate of efficacy for the vaccine (90%, 50-99%) . Follow-up of study children until November 1, 1993, has shown only 1 vaccine failure in an infant, and no invasive infections in those older than 1 year (average age 22 months) . PRP-T vaccine had high protective efficacy with an accelerated immunisation schedule . Furthermore, the vaccine appears to remain protective through the second year of life without a booster dose . These findings provide encouragement for use of PRP-T in the Expanded Programme of Immunisation. South Med J, 1994 Aug, 87(8), 811 - 3 Liver abscess following blunt trauma: a case report and review of the literature; Haight DO et al.; We report a case of Haemophilus paraphrophilus causing primary liver abscesses after blunt nonpenetrating trauma . A 32-year-old previously healthy white man sustained a back injury 2 months prior to admission with fever, chills, and night sweats . A computed tomography (CT) scan-directed needle aspirate of several hypoechoic hepatic lesions grew H paraphrophilus . Recent blunt trauma to the lower back may have contributed to the localization of this infection to an area of contusion or hematoma within the liver, followed by an episode of bacteremia that seeded the injury. Am J Respir Crit Care Med, 1994 Aug, 150(2), 448 - 54 Bronchoalveolar lavage findings in cystic fibrosis patients with stable, clinically mild lung disease suggest ongoing infection and inflammation; Konstan MW et al.; To determine the extent of airway infection and inflammation in adolescents and adults with cystic fibrosis (CF) who have mild lung disease and are without symptoms of active infection, we performed bronchoalveolar lavage (BAL) on 18 CF patients > or = 12 yr of age who were stable, appeared clinically well, and had mean (+/- SEM) FEV1 of 79 +/- 4% of predicted . We quantitated the bacteria, inflammatory cells, immunoglobulins, and mediators of inflammatory tissue damage in the epithelial lining fluid (ELF) of these patients and in 23 healthy control subjects . All CF patients were found to be infected with Pseudomonas aeruginosa, Staphylococcus aureus, and/or Haemophilus influenzae; no organisms were isolated from the control subjects . The mean number of cells in the ELF was 14 times greater in the CF patients than in the control subjects . Neutrophils constituted 57% of the recovered cells in the CF patients versus 3% in the control subjects, and their concentration was 380 times greater in the CF patients versus the control subjects . IgG, IgA, and IgM were 2.5 to 6 times greater in CF ELF versus that of control subjects . Abundant active elastase was present in the ELF of the CF patients (2.3 +/- 0.9 microM) despite threefold elevated levels of alpha 1-protease inhibitor (alpha 1-PI) . No active elastase was detectable in the control subjects . alpha 1-PI was functional in CF as demonstrated by elevated elastase:alpha 1-PI complex (0.045 microM in CF versus 0.002 microM in control subjects) . This active elastase caused proteolytic destruction of surface complement receptors on airway neutrophils in situ.(ABSTRACT TRUNCATED AT 250 WORDS) J Med Microbiol, 1994 Aug, 41(2), 120 - 6 Analysis by pulsed-field gel electrophoresis of insertion mutations in the transferrin-binding system of Haemophilus influenzae type b; Curran R et al.; A mutagenesis system involving the insertion of a non-transposable antibiotic resistance gene cassette was used to generate stable mutations in the chromosome of Haemophilus influenzae type b strain Eagan . The mutations generated were shown by pulsed-field gel electrophoresis (PFGE) to have unique SmaI fi