Acta Microbiol Pol, 1996, 45(1), 19 - 30 Physical maps of the chromosomes of Haemophilus influenzae Sb; Kauc L et al.; The physical maps the Haemophilus influenzae Sb genomes were constructed by the comparison of restriction fragment sizes of complete single and double digests, as well as by the analysis of partial DNA digests . Sites for four restriction endonucleases NotI, RsrII, SmaI and SrfI were located on the bacterial chromosomes, which are circular, and 2028 and 2045 kb in circumferences . The order of fragments was deduced from fragment patterns of the combinations of double digestion and by two-dimensional Field-Inversion Gel Electrophoresis (FIGE). Scand J Infect Dis, 1996, 28(2), 165 - 9 The impact of Haemophilus influenzae type b vaccination in Sweden; Garpenholt O et al.; The number of patients with meningitis and bacteremia due to Haemophilus influenzae was studied in Sweden over the period 1987-1994 . Conjugated H . influenzae type b vaccines were introduced in Sweden in 1992, and all children born after December 31, 1992, were offered vaccination free of charge . A rapid decline of H . influenzae meningitis and bacteraemia was observed in the autumn of 1993, when the expected peak incidence failed to appear . In the prevaccination period 1987-1991, the average annual incidence (cases/100,000) was 34.4 in children aged 0-4 years . In 1994, the annual incidence fell to 3.5 . No significant decline was observed in older children or adults . There was a 92% reduction in the number of meningitis cases and an 83% reduction in cases of bacteraemia . A similar decline was noted in 2 regions which followed different strategies for the introduction of the vaccination programme. Acta Biochim Pol, 1996, 43(1), 107 - 14 Mutagenesis and ultraviolet inactivation of transforming DNA of Haemophilus influenzae complexed with a Bacillus subtilis protein that alters DNA conformation; Setlow JK et al.; The wild-type Bacillus subtilis spore protein, SspCwt, binds to DNA in vitro and in vivo and changes the conformation of DNA from B to A . Synthesis of the cloned SspCwt gene in Escherichia coli also causes large increases in mutation frequency . Binding of SspCwt to transforming DNA from Haemophilus influenzae made the DNA resistant to ultraviolet (UV) radiation . The mutant protein, SspCala, which does not bind DNA, did not change the UV resistance . The UV sensitivity of the DNA/SspCwt complex was not increased when the recipients of the DNA were defective in excision of pyrimidine dimers . These data indicate that the H . influenzae excision mechanism does not operate on the spore photoproduct formed by UV irradiation of the complex . Selection for the streptomycin- or erythromycin-resistance markers on the transforming DNA evidenced significant mutations at loci closely linked to these, but not at other loci . SspCwt apparently entered the cell attached to the transforming DNA, and caused mutations in adjacent loci . The amount of such mutations decreased when the transforming DNA was UV irradiated, because UV unlinks linked markers. Infect Agents Dis, 1996 Jan, 5(1), 8 - 20 Impact of immunization on Haemophilus influenzae type b disease; Madore DV; Epidemiological surveillance programs have shown that before the introduction of effective vaccines, Haemophilus influenzae type b (Hib) was the primary pathogen associated with bacterial meningitis in children . Vaccines composed of the bacterium's polysaccharide conjugated onto protein carriers began to be introduced into routine health care practices for infants as early as 1989 in some European countries . Continued introduction in industrialized nations, including the United States in late 1990, has resulted in the rapid decline in the incidence of reported invasive Hib disease . Follow-up surveillance studies show that (a) the decline in the incidence of Hib disease is temporally related to the introduction of effective vaccines, (b) the decline in Hib epiglottitis preceded the decline in meningitis in the United States, (c) the incidence of disease declined in children under the age of 5 years but remained constant in older children and adults, (d) other bacterial pathogens are now the primary causative agents of infant meningitis and epiglottitis even though the incidence of disease caused by these other pathogens has not changed, and (e) the pharyngeal carriage rate of Hib in children has declined without any evidence of an increase in the carriage of non-type b strains or other pathogens . The introduction of effective conjugate vaccines appears to protect at-risk children from invasive Hib disease as well as reduce the opportunities for interpersonal transmission of this bacterium . In addition, Hib conjugate vaccine utilization has benefited society through economic savings. Dev Biol Stand, 1996, 86, 283 - 96 Development of a guinea-pig model for potency/immunogenicity evaluation of diphtheria, tetanus acellular pertussis (DTaP) and Haemophilus influenzae type b polysaccharide conjugate vaccines; Gupta RK et al.; We have evaluated a guinea pig model for assessing the immunogenicity of Haemophilus influenzae type b (Hib) polysaccharide-protein conjugate vaccines, acellular pertussis vaccine and combination vaccines-consisting of tetanus toxoid (TT), diphtheria toxoid (DT), acellular pertussis vaccine and Hib-TT (Hib-T) conjugate vaccine . The model was based on the United States (US) potency test for TT and DT which requires injection of guinea pigs with a single dose of undiluted vaccine . Guinea pigs showed dose-dependent antibody responses to pertussis toxoid (PTxd) and filamentous haemagglutinin (FHA), two important components of acellular pertussis vaccine . Antibody response of guinea pigs to commercially available Hib conjugate vaccines qualitatively resembled those of human infants . Unconjugated polyribosylribitolphosphate (PRP) was not immunogenic; PRP-D conjugate produced a low antibody response, HbOC, PRP-T (Merieux) and Hib-T (MPHBL) produced a low response to the first dose and a strong anamnestic response to the booster dose . PRP-OMP uniquely produced a strong response after the first dose which was boosted by the second dose . In preliminary experiments, injection of guinea pigs with the combined vaccine formulations consisting of TT, DT, whole cell or acellular pertussis vaccine (Ptxd and FHA) and Hib-T conjugate showed that these vaccines were immunogenic when combined, with some effects on the antibody responses of certain components . This model for testing potency/immunogenicity of combined vaccines substantially reduces the number of animals needed to test each lot of vaccine . To reduce the use of animals in testing vaccines further, we propose the use of a Vero cell assay for titrating diphtheria antitoxin and ELISA for measuring IgG antibody to tetanus toxin . The guinea pig model may also be useful for evaluating combination vaccines. Clin Diagn Lab Immunol, 1996 Jan, 3(1), 84 - 8 Interlaboratory study evaluating quantitation of antibodies to Haemophilus influenzae type b polysaccharide by enzyme-linked immunosorbent assay; Madore DV et al.; An interlaboratory study was conducted to determine whether an enzyme-linked immunosorbent assay (ELISA) with an antigen preparation composed of various-sized fragments of Haemophilus influenzae type b polysaccharide conjugated to human serum albumin could be standardized across laboratories and whether the ELISA-derived results from different laboratories are equivalent to those obtained by the standard radioactive antigen binding assay (RABA) for quantitation of anti-H, influenzae type b polysaccharide antibodies . Twenty coded human serum samples were quantitated by ELISA in 11 laboratories and by RABA in 5 laboratories . The mean RABA-derived values served as the basis for all comparisons . While the overall correspondence of antibody values between the two methods was good, significant differences were found among some of the 11 ELISA data sets and among the mean RABA values . Seven laboratories generated higher ELISA antibody values for low-titered sera . Four laboratories generated antibody concentrations that were not statistically different between the two assay methods . The results therefore indicate that the ELISA can tolerate substantial variations in protocol, such as the use of different plates and different antibody reagents, without affecting the quantitation of serum antibodies . However, attention should be focused on low-titered sera, as some assay conditions may yield spurious results . This ELISA is a serologic assay which can serve as an alternative to the RABA for quantitation of antibodies to H . influenzae type h polysaccharide. Clin Diagn Lab Immunol, 1996 Jan, 3(1), 37 - 41 Effect of malnutrition on serum and milk antibodies in Zairian women; Brussow H et al.; Serum and human milk antimicrobial antibody titers were measured longitudinally in 17 malnourished and 14 control Zairian women during 6 to 18 months of lactation to test whether malnutrition is specifically associated with an impaired secretory antibody response . No decreases in total serum and human milk immunoglobulin concentrations, neutralizing antibody titers against rotavirus, or specific enzyme-linked immunosorbent assay antibody titers against rotavirus, respiratory syncytial virus, Escherichia coli, Streptococcus pneumoniae, and Haemophilus influenzae were detected when malnourished women were compared with control women . Malnutrition had no effect on circulating and secretory antibody concentrations in Zairian women . Daily human milk outputs, however, were about 30% lower in malnourished than in control women, resulting in a correspondingly lower ingestion of immunoglobulins by the children of malnourished women. Chemotherapy, 1996 Jan-Feb, 42(1), 71 - 77 Haemophilus influenzae type b exoproducts induce chemotaxis and macrolide antibiotic release by human polymorphonuclear leukocytes; Fontan PA et al.; The capacity of phagocytes to concentrate macrolide antibiotics was suggested by previous reports . In this study, we evaluated the capacity of Haemophilus influenzae type b culture supernatants (HICS) to induce polymorphonuclear leukocyte (PMNL) migration and macrolide antibiotic delivery . Using a Boyden multiwell chamber and a chemotaxis assay under agarose combined with a bioassay to measure antibiotic levels in agar, we demonstrated the chemotactic activity of HICS . Preincubation of PMNL with either erythromycin or azithromycin did not affect PMNL chemotaxis . By the agar diffusion test, we established that HICS increased the release of antibiotic from PMNL when compared with spontaneous release . Furthermore, we determined that the antibiotics remain bioactive after release . These results suggest that HICS may have a modulatory effect on transport and delivery of macrolide antibiotics by PMNL at the infection site. J Clin Microbiol, 1996 Jan, 34(1), 49 - 54 Simultaneous PCR detection of Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus types 1 and 2 from genital ulcers; Orle KA et al.; A multiplex PCR (M-PCR) assay with colorimetric detection was devised for the simultaneous amplification of DNA targets from Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus (HSV) types 1 and 2 . By using target-specific oligonucleotides in a microwell format, 298 genital ulcer swab specimens collected in New Orleans during three intervals from 1992 through 1994 were evaluated . The results of the M-PCR assay were compared with the results of dark-field microscopy and H . ducreyi culture on two different culture media . HSV culture results were available for 99 specimens collected during the third interval . Confirmatory PCR assays targeting different gene sequences for each of the three organisms were used to validate the M-PCR results . Specimens were resolved as positive for the determination of sensitivity if the reference diagnostic test was positive or if the results of both the M-PCR and the confirmatory PCR were positive . The resolved sensitivities of M-PCR for HSV, H . ducreyi, and T . pallidum were 100, 98.4, and 91%, respectively . The resolved sensitivities of HSV culture, H . ducreyi culture, and dark-field microscopy were 71.8, 74.2, and 81%, respectively . These results indicate that the M-PCR assay is more sensitive than standard diagnostic tests for the detection of HSV, H . ducreyi, and T . pallidum from genital ulcers. Eur Arch Otorhinolaryngol, 1996, 253(4-5), 260 - 3 The nasopharyngeal bacterial flora in children with otitis media with effusion; Fujimori I et al.; A study was undertaken to evaluate the incidence of nasopharyngeal alpha-streptococci with inhibitory activity against pathogens, as a defense mechanism of the normal bacterial flora against infection . Cultures were taken from the nasopharynges of 43 children with otitis media with effusion (OME) . The detection rates of alpha-streptococci with inhibitory activity against Haemophilus influenzae, Streptococcus pneumoniae, Staphylococus aureus and group A streptococci were significantly lower in the nasopharynx than those isolated from the tonsils of the same patients . Moreover, the detection rates of alpha-streptococci with inhibitory activity against all of these pathogens derived from the nasopharynx were lower than those in healthy children, streptococcal strains with activity against H . influenzae and Strep, pneumoniae were also lower than that in patients with tonsillitis . These findings suggest that low nasopharyngeal levels of alpha-streptococci with inhibitory activity against respiratory pathogens may render children susceptible to OME . Further studies are needed to investigate the relationships between the prevalence of pathogens in the nasopharynx and the inhibitory activities of alpha-streptococci against them in order to devise and select optimal treatment for patients with OME. Am J Vet Res, 1996 Jan, 57(1), 63 - 7 Detection of strain-specific antigenic epitopes on the lipo-oligosaccharide of Haemophilus parasuis by use of monoclonal and polyclonal antibodies; Zucker BA et al.; OBJECTIVE: To investigate the antigenic diversity of lipo-oligosaccharides of Haemophilus parasuis . PROCEDURES: Immunoblot assays were done with monoclonal and polyclonal antibodies on whole-cell lysates . Individual colonies of H parasuis strains H 54, H 53, and H 128 were tested for reactivity with lipo-oligosaccharide-specific monoclonal antibodies after a single passage on chocolate agar, and colonies of strain H 54 were analyzed after 10 passages . Colony blot tests were used to screen H parasuis strains for spontaneously occurring antigenic variation in their lipo-oligosaccharides . RESULTS: Eight H parasuis strains were separated into 4 lipo-oligosaccharide serovars on the basis of immunoblot reactions with 3 polyclonal rabbit antisera . Nine monoclonal antibodies against lipo-oligosaccharides of a lipo-oligosaccharide-serovar I strain reacted with all tested serovar I strains but failed to react with other H parasuis strains . CONCLUSIONS: Variations in the antigenic reactivity after 1 or 10 passages on chocolate agar were not observed . The serovar I lipo-oligosaccharide strains included virulent as well as avirulent H parasuis strains, indicating that these epitopes do not correlate directly with virulence properties of H parasuis. Rev Mal Respir, 1996, 13(2), 175 - 82 {Acute community-acquired pneumonia of moderate and grave severity investigated by bronchoscopy . Analysis of 193 cases hospitalized in a general hospital}; Vives L et al.; Between February 1989 and June 1994 193 cases of acute community acquired pneumonia (PAC) which were of intermediate or great severity were admitted to two hospitals in the South West of France . These patients were explored using bronchofibroscopy (FB) with a protected brush (BP) and alveolar microlavage (MLBA) and quantitative cultures were performed, also there were other specimens taken in a regular fashion . The percentage of positive examinations was 60% for brushings (BP), 59% for MLBA and 21% for blood cultures and 16% for serological tests . An aetiology was determined in 137 cases (70.9%) . The organisms recovered were Streptococcus pneumoniae (49.6%), gram negative bacilli (17.4%), Haemophilus influenzae (11.7%), Mycoplasma pneumoniae (4.4%), Mycobacterium tuberculosis (4.4%), Staphylococcus aureus (3.6%), Chlamydia pneumoniae (2.2%), Legionella pneumophila (0.7%), and various 5.8% . The overall mortality was 15% despite immediate antibiotics based on the likely organism in 88% of cases . The study of prognostic factors confirmed the Fine score system (determined a posteriori) which constitutes a useful and practical index determining the management of PAC . On the other hand the role of bacteriological documentation in improving the vital prognosis remains to be confirmed . If bronchofibroscopy has appeared to us as a safe and useful means of investigation, the management of these disease remains to specified . We suggest that its use is reserved for subjects with life threatening disease (a Fine score equal to or greater than 3) or for those patients who are likely to have unusual germs: failure of previous antibiotics, diabetes, malnourishment, cancer, airflow obstruction and inhalation. Arch Med Res, 1996 Summer, 27(2), 133 - 8 Experimental infection by Haemophilus influenzae type b in inbred mice; Sifontes S et al.; The epidemiological situation of bacterial meningitis is increasing dramatically . There is no doubt that the lack of proper animal models has hampered the achievement of effective prophylactic and therapeutic means . We report the characterization of the experimental disease caused by Haemophilus influenzae type b (Hib) in mice, taking into account its importance as an etiological agent of such a type of meningitis . The high resistance of C57BL/6, CBA/ J and BALB/cJ mice to Hib infection was proven . LD50 of Hib using trypsin or iron dextran as virulence enhancement factors (VEF), both being similar and more than 1000 times lower than that without any VEF, were determined . Lesions of CNS compatible with meningitis were found in about one third of specimens . Hair bristling, conjunctivitis, diarrhea, depression and prostration were the most characteristic symptoms . The proportion of animals which die is highest on the first day, lower on the second and almost zero after 48 h of infection . Water and food intake was higher in control than in infected animals; nevertheless, there were no differences in body weight increase among the mice after 5 days post-infection . Microorganisms were isolated from CSF and blood after 6 h of infection and positive results remained according to the size of the inoculum . Despite the acuteness of the experimental disease, antibiotic treatment with internationally recommended drugs was shown to be effective . Similar results were achieved when hyperimmune serum vs . Hib was applied. Pediatr Infect Dis J, 1996 Jan, 15(1), 58 - 61 Etiology of acute otitis media in human immunodeficiency virus-infected children; Marchisio P et al.; BACKGROUND: Acute otitis media (AOM) is one of the most common infections that are implicated as significant contributors to morbidity in HIV-infected children . To establish the optimal antibiotic therapy tympanocentesis is indicated as the first line diagnostic procedure, because unusual pathogens may play a role in advanced stages of deficient humoral or cellular immunity . METHODS: The microbiology of 60 episodes of AOM diagnosed in 21 symptomatic HIV-infected children (ages 9 months to 12 years) was compared with that of 121 episodes of AOM occurring in 113 immunocompetent HIV-negative children (ages 6 months to 12 years) in the last 5 years . RESULTS: The prevalence of the three most common pathogens (Streptococcus pneumoniae, Haemophilus influenzae and group A beta-hemolytic Streptococcus) was similar in HIV-infected and in normal children (56.5% vs . 54.9% of the ears) . Staphylococcus aureus was significantly more frequent in AOM diagnosed in severely immunosuppressed stages . A significantly lower proportion of middle ear effusions obtained in HIV-infected children yielded no bacteria compared with normal children . Beta-lactamase production among isolates of H . influenzae was a rare phenomenon, both in HIV-infected and in normal children . No penicillin-resistant S . pneumoniae was found . CONCLUSIONS: In HIV-infected children with absent or moderate immunosuppression empiric antibiotic therapy should be based on the recommendations given for immunocompetent children of the same geographic area . In severe immunosuppressed stages, given the possible role of Staph . aureus, extended spectrum antibiotics should be considered. J Am Vet Med Assoc, 1996 Jan 1, 208(1), 102 - 6 Comparison of tilmicosin with long-acting oxytetracycline for treatment of respiratory tract disease in calves; Musser J et al.; OBJECTIVE--To compare the effect of a single parenteral injection of tilmicosin with that of a single dose of a long-acting oxytetracycline as treatment in the early stages of naturally acquired undifferentiated respiratory tract disease in young dairy calves . DESIGN--Prospective clinical trial, randomized block design . ANIMALS--40 dairy calves . PROCEDURES--78 calves from 5 farms were examined weekly until 3 months old . When respiratory tract disease was diagnosed by a veterinarian, the calf was assigned to 1 or 2 treatment groups . Transtracheal wash samples were acquired to characterize the pathogens . The veterinarian, who was unaware of treatment assignments, examined calves for 3 days after treatment and evaluated severity, using a scoring system . Growth rates were measured . RESULTS--On the basis of response to initial treatment, relapse rates, and effect on growth rates, the antibiotics were determined to be equally effective . Severity of clinical disease was significantly (P < 0.03) less for the tilmicosin-treated calves on days 2 and 3 after treatment . Findings from analysis of transtracheal wash samples indicated Pasteurella multocida (25/40), P haemolytica (4/40), Haemophilus somnus (4/40), Actinomyces pyogenes (3/40), and Aspergillus sp (2/40) . Mycoplasma was isolated in association with bacterial isolates in 22 calves . CLINICAL IMPLICATIONS--Tilmicosin and oxytetracycline are effective in treatment of respiratory tract disease in young calves, even when Mycoplasma spp are involved . Tilmicosin is more effective in resolving clinical signs . Early treatment of dairy calves with respiratory tract disease may decrease detrimental effects on growth. Immunogenetics, 1996, 44(2), 115 - 20 New alleles of IGKV genes A2 and A18 suggest significant human IGKV locus polymorphism; Atkinson MJ et al.; The human kappa light chain consists of approximately 35 potentially functional IGKV genes . However, an estimation of the diversity in the IGKV repertoire of an individual will be affected by the extent of polymorphisms for the different IGKV genes and their patterns of inheritance . To date, little information is available to indicate the extent of allelic variation of the IGKV genes . We examined the extent of allelism for one IGKV gene pair, the distal region A2 gene and its closely related proximal region duplicate A18 . We found two new alleles for A2 and one new allele for A18, and sequenced approximately 1 kilobase flanking each gene . The new A18 allele, unlike the originally described allele, appears to be functional . All these alleles were found at relatively high frequencies in the four ethnic populations studied, with the exception of the defective A2b allele which was highly represented only in Navajos . The originally described A2a allele encodes for the predominant protective antibody against Haemophilus influenzae . Therefore, the patterns of allelic inheritance described for this IGKV gene pair indicate that allelism in the IGKV locus is likely to have a significant impact on immune responses. Int J STD AIDS, 1996, 7 Suppl 1, 9 - 12 Treatment of chancroid with azithromycin; Ballard RC et al.; A randomized, comparative study undertaken in Nairobi, Kenya and a non-comparative evaluation undertaken in Carletonville, South Africa have both shown that a single oral dose of azithromycin 1 g is effective in the treatment of the genital ulcer disease (GUD), chancroid, with cure rates of 89% and 92% recorded respectively . While treatment failure was associated with human immunodeficiency virus seropositivity and lack of circumcision in Kenya, no such association could be found in the South African study . In both series, azithromycin treatment resulted in cure of both Haemophilus ducreyi culture-positive and culture-negative cases of GUD, including two cases subsequently diagnosed as lymphogranuloma venereum . A combination of single-dose azithromycin with single-dose benzathine penicillin may provide effective 'single-visit' syndromic treatment for GUD in many developing countries. APMIS, 1996 Jan, 104(1), 61 - 7 Basophil-bound IgE and serum IgE directed against Haemophilus influenzae and Streptococcus pneumoniae in patients with chronic bronchitis during acute exacerbations; Kjaergard LL et al.; The investigation includes 12 patients hospitalized with acute exacerbations of chronic bronchitis (CB) and infected in the lower respiratory tract with Haemophilus influenzae (HI) or Streptococcus pneumoniae (SP) . Eight patients were infected the HI, three with SP, and one patient with both species . Basophil-bound IgE and serum IgE directed against these species were examined using the patients' own bacterial isolates . All patients showed IgE-mediated histamine release when their peripheral leukocytes were incubated in vitro with the infecting species, indicating basophil-bound IgE directed against their own bacterium . No IgE-mediated response was obtained in the control group of 12 healthy individuals . Bacteria-specific IgE in serum was demonstrated by immunofluorescence assay and further verified by passive sensitization . There was a positive serum titre in seven of nine patients housing HI and in all SP-infected patients but not in the control group . No synchronism was found between a positive response in the histamine release test and the immunofluorescence assay by parallel testing during the test period . This may be due to a time delay between production of serum IgE and its fixation to the cell surface . The results indicate a potential for a bacteria-specific IgE-mediated immune response in CB . Thus, by triggering mediator release, bacteria may be involved in the pathogenesis of exacerbations in CB. J Pediatr, 1996 Jan, 128(1), 102 - 3 Invasive Haemophilus influenzae type b infection in a child with familial deficiency of the beta subunit of the eighth component of complement; Pallares DE et al.; A child who had had meningitis caused by Haemophilus influenzae type b, and then had meningococcal meningitis, was found to have familial deficiency of the beta subunit of the eighth component of complement . The child had not received the H . influenzae type b vaccine . If this deficiency is discovered, we recommend that family members be screened, regardless of their health status. J Bacteriol, 1996 Jan, 178(2), 564 - 70 The lipooligosaccharides of Haemophilus ducreyi are highly sialylated; Melaugh W et al.; The major lipooligosaccharides of the sexually transmitted pathogen Haemophilus ducreyi 35000 have been previously found to terminate in N-acetyllactosamine and sialyl-N-acetyllactosamine, Neu5Ac alpha 2-->3Gal beta 1-->4GlcNAc (W . Melaugh, N . J . Phillips, A . A . Campagnari, M . V . Tullius, and B . W . Gibson, Biochemistry 33: 13070-13078, 1994) . In this study, mass spectrometry and composition analyses have shown that the lipooligosaccharides from three other H . ducreyi strains also contain N-acetyllactosamine and are highly sialylated (approximately 30 to 50%), although one African strain was found to contain neither of these structural features. J Bacteriol, 1996 Jan, 178(2), 396 - 402 Altered lipopolysaccharide characteristic of the I69 phenotype in Haemophilus influenzae results from mutations in a novel gene, isn; Preston A et al.; The 169 phenotype of Haemophilus influenzae results from a mutation leading to a lipopolysaccharide molecule consisting only of lipid A and a single phosphorylated 2-keto-3-deoxyoctulosonic acid residue . In this paper we describe the identification of a gene which, when mutated, results in the 169 phenotype . We have named the gene isn . The predicted amino acid sequence of Isn is homologous to the product of the lmbN gene involved in the biosynthesis of the sugar-containing antibiotic lincomycin by Streptomyces lincolnensis . lsn is situated between two loci that are homologous to the dpp and art periplasmic permease systems in Escherichia coli . Northern (RNA) blot and primer extension analyses reveal that isn is transcribed as a monocistronic mRNA . Potential functions of Isn protein are discussed. J Infect Dis, 1996 Jan, 173(1), 180 - 6 A virulent nonencapsulated Haemophilus influenzae; Nizet V et al.; Nontypeable Haemophilus influenzae strain INT1 was isolated from the blood of a young child with clinical signs of meningitis following acute otitis media . No immunologic or anatomic predisposition of this child for invasive bacterial infection with an unusual organism was documented . Sensitive ELISA proved the absence of intra- or extracellular capsular polysaccharide production by INT1 and Southern blot analysis confirmed the lack of an intact capsulation (cap) gene locus within the chromosome . Nevertheless, INT1 established bacteremia and meningitis in infant and weanling rat models of invasive H . influenzae infection . High-molecular-weight DNA isolated from INT1 was shown to confer an invasive phenotype on transformation of a nonencapsulated, avirulent laboratory strain of H . influenzae . Together these findings imply the presence of one or more as-yet-undiscovered, noncapsular virulence factors of H . influenzae that are capable of mediating invasive disease and resistance to immunologic clearance. Am J Med, 1995 Dec 29, 99(6B), 3S - 7S Trends in antimicrobial susceptibility of bacterial pathogens of the respiratory tract; Doern GV; Rates of antimicrobial resistance have been increasing in bacteria responsible for community-acquired lower respiratory tract infections in the United States . Nearly 100% of clinical isolates of Moraxella catarrhalis now produce beta-lactamase, an enzyme that renders this pathogen resistant to such agents as penicillin, ampicillin, and amoxicillin . However, this organism remains nearly uniformly susceptible to alternative oral antimicrobials, such as cephalosporins, macrolides, tetracyclines, beta-lactamase inhibitor combinations, and the combination of trimethoprim/sulfamethoxazole . The susceptibility of M . catarrhalis to these agents is not expected to change markedly in the next few years . A linear increase in the prevalence of beta-lactamase-mediated ampicillin resistance has been evident among isolates of nontypeable Haemophilus influenzae during the past decade in the United States . By the year 2000, 45-50% of isolates are likely to produce beta-lactamase . Although the susceptibility of this organism to alternative oral antimicrobials varies, rates of resistance to cefuroxime axetil, cefpodoxime, cefixime, azithromycin, and perhaps clarithromycin remain < 1% . The rate of penicillin resistance among isolates of Streptococcus pneumoniae, which has increased steadily in recent years, currently stands at approximately 25% in the United States and will likely reach 40-50% during the next 5-10 years . Because of cross-resistance, in general all beta-lactam antimicrobials have reduced activity against penicillin-resistant strains of S . pneumoniae . A 1994-1995 survey found that 3.4% of S . pneumoniae isolates were highly resistant to cefotaxime, and 4-8% were resistant to chloramphenicol, tetracycline, and the macrolides . Resistance to these antimicrobials has usually followed the emergence of penicillin resistance in other countries . Therefore, S . pneumoniae resistance to these drugs is expected to increase markedly during the next few years in the United States. J Biol Chem, 1995 Dec 22, 270(51), 30344 - 52 Cloning and expression of the mammalian cytosolic branched chain aminotransferase isoenzyme; Hutson SM et al.; The cDNA for the rat cytosolic branched chain aminotransferase (BCATc) has been cloned . The BCATc cDNA encodes a polypeptide of 410 amino acids with a calculated molecular mass of 46.0 kDa . By Northern blot analysis, BCATc message of approximately 2.7 kilobases was readily detected in rat brain, but was absent from liver, a rat hepatoma cell line, kidney, and skeletal muscle . When expressed in COS-1 cells, the enzyme is immunologically indistinguishable from the native enzyme found in rat brain cytosol . Comparison of the rat BCATc sequence with available data bases identified the Escherichia coli (and Salmonella typhimurium) branched chain aminotransferase (BCAT) and revealed a Haemophilus influenzae BCAT, a yeast BCAT, which is hypothesized to be a mitochondrial form of the enzyme, and the murine BCATc (protein ECA39) . Calculated molecular masses for the complete proteins are 33.9 kDa, 37.9 kDa, 42.9 kDa, and 43.6 kDa, respectively . The rat BCATc sequence was 84% identical with murine BCATc, 45% identical with yeast, 33% identical with H . influenzae, 27% identical with the E . coli and S . typhimurium BCAT, and 22% identical with the evolutionary related D-amino acid aminotransferase (D-AAT) (Tanizawa, K., Asano, S., Masu, Y., Kuramitsu, S., Kagamiyama, H., Tanaka, H., and Soda, K . (1989) J . Biol . Chem . 264, 2450-2454) . Amino acid sequence alignment of BCATc with D-AAT suggests that the folding pattern of the overlapping mammalian BCATc sequence is similar to that of D-AAT and indicates that orientation of the pyridoxal phosphate cofactor in the active site of the eukaryotic BCAT is the same as in D-AAT . Thus, BCAT are the only eukaryotic aminotransferases to abstract and replace the proton on the re face of the pyridoxal phosphate cofactor . Finally, requirements for recognition of substrate L-amino acid and alpha-carboxylate binding are discussed. Pharmacoeconomics, 1996, 9 Suppl 3, 8 - 15; discussion 23-5 Economic evaluation of vaccination; Van Damme P et al.; With increasing expenditures in healthcare, in absolute terms as well as in relative terms, interest in the efficiency of certain interventions in healthcare has also increased . Faced with the limitations of the healthcare budget, budget holders try to find the optimal way of dividing their funds over different healthcare provisions, without discarding human and medical considerations . In economic terms, this process could be called the 'optimal allocation of scarce resources over the inputs of a function of production' . The means of production would then be 'the provision of healthcare', whereas the output would be 'improvement of health' . Clearly choices have to be made with regard to spending the healthcare budget . One of the instruments that can help in making such choices is the economic evaluation . In economic evaluations of vaccinations, different vaccination strategies are defined . The consequences in terms of costs and effects of each strategy are being calculated and compared with a reference strategy, which is often the nonintervention strategy, i.e . 'no vaccination' . According to the way in which the benefit or the output of vaccination-'improvement of health'-is measured, a distinction is made between various methods of economic evaluation: in a cost-effectiveness analysis, health gains are measured in natural units (e.g . prevented infections, prevented illness days, life-years gained, etc.); in a cost-utility analysis, the quality of the health gains is taken into account (e.g . quality-adjusted life-year); and in a cost-benefit analysis, health gains are converted into monetary units . Costs can be divided into direct and indirect costs . Direct costs are directly related to medical treatments (medication, laboratory tests, consultations, etc.) or to vaccination (e.g . purchasing price of the vaccine, costs for administering the vaccine, treatment of side effects, etc.) . Costs indirectly related to treatments and vaccination are mainly costs of lost productivity due to disease morbidity or mortality, and opportunity costs . In comparison with other vaccine-preventable infections, influenza vaccination for the elderly seems acceptable from an economic point of view (about $US650 per life-year gained, in 1981) . Cost-effectiveness ratios of other vaccinations range from about $US720 per life-year gained for universal hepatitis B vaccination to about $US190,000 per life-year gained for universal Haemophilus influenzae type by vaccination . Because of differences in methods, the representation of results, and country-specific parameters, different economic evaluations of the same vaccination strategy may show divergent results . Therefore, until sufficient standardisation of economic evaluations exists, comparisons of the sort we are making here should be interpreted with prudence. J Paediatr Child Health, 1995 Dec, 31(6), 513 - 8 A case-control study examining risk factors for invasive Haemophilus influenzae type b disease in Victoria, Australia 1988-90; Clements DA et al.; OBJECTIVE: To determine whether day-care attendance was a risk factor for Haemophilus influenzae type b (Hib) disease, particularly for epiglottitis . METHODOLOGY: A case-control analysis of risk factors for invasive Hib disease was performed in Victoria, Australia between February 1988 and February 1990 prior to the introduction of immunization for Hib . A total of 210 cases and 367 day surgery hospital controls were enrolled prospectively . Data were collected by questionnaire at the time of admission . RESULTS: Logistic regression analysis showed that risk factors for meningitis were day-care attendance, household crowding and recent illness in a sibling . Risk factors for epiglottitis were day-care attendance and mother's birthplace in Australia or New Zealand . CONCLUSIONS: This study confirms that day-care attendance is a risk factor for Hib epiglottitis as well as meningitis . In addition, the mother's birthplace in Australia or New Zealand is a risk factor for epiglottitis in these data . The reason for this latter observation is unclear. Afr J Med Med Sci, 1995 Dec, 24(4), 353 - 7 Antibiogram of bacterial isolates from cases of otitis media and lower respiratory tract infections; Ndip RN et al.; A total of one hundred and two cases of otitis media were screened for the isolation of bacterial flora of ears . Out of this, Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae accounting for 41.2%, 25.5% and 13.3% respectively were isolated . A further two hundred and four sputum samples from cases of lower respiratory tract infections were screened . Moraxella catarrhalis, S . aureus, S . pneumoniae and H . influenzae constituting 20.9%, 37%, 30% and 21% were isolated in that order . Thirty-one point seven percent of the Moraxella catarrhalis isolates were beta-lactamase positive . Beta-lactamase M . catarrhalis were resistant to penicillin and ampicillin while the non-beta-lactamase producers were sensitive to these antimicrobial agents . However, both beta-lactamase producers and non-producers were resistant to trimethoprim but sensitive to erythromycin, tetracycline and amoxycillin . Staphylococcus aureus, S . pneumoniae and H . influenzae was also sensitive to penicillin while S . pneumoniae was also sensitive to erythromycin and H . influenzae to chloramphenicol. Microb Pathog, 1995 Dec, 19(6), 391 - 407 Cloning and molecular analysis of the Isi1 (rfaF) gene of Neisseria meningitidis which encodes a heptosyl-2-transferase involved in LPS biosynthesis: evaluation of surface exposed carbohydrates in LPS mediated toxicity for human endothelial cells; Jennings MP et al.; Neisseria meningitidis, but not Haemophilus influenzae, damage cultured human endothelial cells . We have undertaken a study to generate genetically and structurally defined lipopolysaccharide (LPS) mutant strains of meningococci for functional studies to assess the role of surface exposed oligosaccharides in imparting specificity of toxic damage to human endothelial cells . The Isi1 gene, which had been shown to be involved in LPS biosynthesis of Neisseria gonorrhoeae, was amplified by PCR and cloned . Nucleotide sequence analysis confirmed the identity of the clone and revealed homology with Isi1 of N . gonorrhoeae and the rfaF gene of Salmonella typhimurium which encodes a heptosyl-2-transferase involved in LPS biosynthesis . The identity of the cloned Isi1 gene, as a functional rfaF homologue, was confirmed by the complementation of a S . typhimurium rfaF mutant using a P22 phage sensitivity test . An Isi1 mutant meningococcal strain was constructed, and structural analysis of the mutant LPS molecule revealed a single heptose in the core structure, consistent with a heptosyl-2-transferase deficient mutant . In order to investigate the relative cytotoxicities of meningococci expressing native and altered LPS, wild type, Isi1, and galE strains were compared in cytotoxicity assays using human umbilical vein endothelial cells (Huvecs) in culture . Analysis using Huvecs derived from several individuals (cords) showed that the three phenotypes were almost equally cytotoxic . Removal of the terminal portion (galE mutant) or the majority (Isi mutant) of the oligosaccharide did not effect LPS-mediated cytopathic damage to Huvecs in a culture suggesting that the oligosaccharide portion did not play a major role in cytotoxicity. Mol Microbiol, 1995 Dec, 18(5), 975 - 86 The product of the pilQ gene is essential for the biogenesis of type IV pili in Neisseria gonorrhoeae; Drake SL et al.; The product of the Neisseria gonorrhoeae omc gene possesses regions homologous to those found in members of a protein superfamily that are associated with the translocation of proteins and DNA-protein complexes across the outer membrane . Amongst its protein homologues, Omc has higher overall homology to PilQ, which is required for type IV pilus expression in Pseudomonas aeruginosa, and OrfE, which is required for sequence-specific DNA uptake by Haemophilus influenzae . The function of Omc, however, is unknown and gonococcal omc mutants have not been described . We constructed gonococcal mutants expressing truncated forms of the protein, and found that these mutants are severely defective for both pilus expression and competence for natural transformation . To be consistent with pre-existing pilus gene nomenclature, we have redesignated the gene pilQ instead of omc, and its product, PilQ instead of Omc . The MS11 gene was sequenced and found to differ from the DNA sequence reported for that of another gonococcal strain; these differences were associated with a repeated DNA element, suggesting a genetic basis for structural variation in PilQ . The results also show that PilQ- mutants are distinct from previously described gonococcal pilus-assembly mutants and P . aeruginosa PilQ- mutants by virtue of their expression of rare pilus filaments . Taking these data into account, PilQ is proposed to function in the terminal steps of organelle biogenesis by acting as a pilus channel or pore. Mol Microbiol, 1995 Dec, 18(5), 821 - 30 Cloning and characterization of the genes encoding the hemolysin of Haemophilus ducreyi; Palmer KL et al.; We previously identified a heat- and protease-labile haemolytic activity expressed by Haemophilus ducreyi . In order to characterize the haemolysin at the molecular level, genomic DNA from H . ducreyi was probed with haemolysin genes from other Gram-negative organisms . The haemolysin genes of Proteus mirabilis hybridized to H . ducreyi DNA suggesting that the haemolysin of H . ducreyi is related to the Proteus/Serratia pore-forming family of haemolysins . Tn916 mutagenesis was employed to isolate haemolysin-deficient mutants . Approximately 5000 Tn916 transposon mutants were screened for the loss of haemolytic activity and two mutants were identified . One mutant, designated 35,000-1, was further characterized . Sequences flanking the Tn916 element in strain 35,000-1 were employed to identify clones from a lambda DASHII library of H . ducreyi strain 35,000 DNA . A 13 kb insert from one lambda clone was selected for further study . This 13 kb fragment was able to both confer haemolytic activity to Escherichia coli and complement the haemolysin deficiency in strain 35,000-1 . The haemolysin gene cluster was cloned from this 13 kb insert and two genes, designated hhdA and hhdB, were identified . The derived amino acid sequence of these genes demonstrated homology to the haemolysin and activation/secretion proteins of P . mirabilis and Serratia marcescens. Clin Infect Dis, 1995 Dec, 21 Suppl 3, S218 - 25 The management of pneumonia in children in developing countries; Shann F; Pneumonia kills about 3 million children every year in developing countries, and it is now clear that most fatal pneumonia is caused by Haemophilus influenzae or Streptococcus pneumoniae . To reduce mortality associated with pneumonia, the World Health Organization has developed guidelines for the treatment of children in developing countries who have cough or difficulty breathing: children without tachypnea or chest indrawing do not need antibiotic therapy; children with tachypnea but no chest indrawing should have antibiotic therapy at home; and children with chest indrawing should be admitted to the hospital for intramuscular injections of benzylpenicillin or chloramphenicol . Universal application of these guidelines would save the lives of approximately 600,000 children every year . Other important issues are oxygen therapy, fluid restriction, limitation of the use of acetaminophen, pneumonia in neonates, and the emergence of antibiotic resistance . There is an urgent need for vaccines that protect infants against infection with S . pneumoniae and all strains of H . influenzae, including nonserotypeable strains. Clin Infect Dis, 1995 Dec, 21(6), 1471 - 3 Childhood bacterial meningitis in Southwestern Greece: a population-based study; Syrogiannopoulos GA et al.; During the 5-year period from 1990 through 1994, we investigated the incidence of childhood bacterial meningitis (beyond the neonatal period) in southwestern Greece . Thirty-seven cases of bacterial meningitis were identified . Fifty-one percent of the cases were caused by Neisseria meningitidis, 41% by Haemophilus influenzae type b (Hib), and 8% by Streptococcus pneumoniae . Fifty-eight percent and 73% of cases of meningococcal and Hib meningitis, respectively, involved children younger than 2 years of age . The average yearly incidences of meningococcal and Hib meningitis were 9.7 and 8 cases, respectively, per 100,000 children < 5 years of age . The main difference between findings in this study and those in other studies in Western European countries is the lower incidence of Hib meningitis observed in our area . This low incidence of Hib meningitis cannot be attributed to the use of vaccine because the Hib vaccine was first introduced in May 1994 and used infrequently through December 1994. Eur J Oral Sci, 1995 Dec, 103(6), 382 - 7 Effect of titanium on selected oral bacterial species in vitro; Leonhardt A et al.; Titanium granules were tested for their antibacterial effect on strains of Streptococcus sanguis, Streptococcus mitis, Actinomyces naeslundii, Haemophilus parainfluenzae, Fusobacterium spp . and Prevotella intermedia in comparison with amalgam and two of its components, copper and tin . Glass beads were used as controls . The number of viable bacteria was estimated in samples exposed to the various materials for 1, 3, 6 and 24 h, respectively, and the viable counts were related to the baseline value . Titanium showed low antibacterial effect on the species tested . Copper and amalgam showed an expressed toxicity to all species and differed significantly from titanium and glass particles . Gram positive Streptococcus spp . and A . naeslundii showed a lower susceptibility to the metals than the Gram negative species . The antibacterial effect of copper and amalgam test particles on S . sanguis and P . intermedia was significantly decreased in the presence of serum . This study showed that some metals have a toxic effect in vitro on oral bacteria, a fact that may play a role in plaque formation when these materials are used for dental restorations . Titanium did not have a similar antibacterial effect. Pediatr Infect Dis J, 1995 Dec, 14(12), 1075 - 8 Increasing incidence of penicillin- and ampicillin-resistant middle ear pathogens; Rodriguez WJ et al.; During a 13-month period ending in January, 1995, we obtained 159 samples of middle ear exudate through tympanocentesis (n = 155) or acute spontaneous otorrhea (n = 4) from 151 children enrolled in therapeutic trials of acute otitis media in a pediatric practice in Northern Virginia . Their ages ranged from < 1 to > 6 years of age (mean, 35 months; median, 22 months) . Precise diagnostic criteria for acute otitis media always included bulging outward of all or part of the eardrum, opacification of the eardrum regardless of color and impaired mobility to positive and negative pressure via the pneumatic otoscope . Bacterial pathogens were isolated from middle ear fluid in 95% of these children: Streptococcus pneumoniae was recovered from 61 (37%); Haemophilus influenzae from 45 (27%); Moraxella catarrhalis from 41 (25%); Group A streptococcus from 6 (4%); Staphylococcus aureus from 4 (2%); and no growth or microbes of uncertain significance from 8 (5%) . Six of the patients had mixed bacterial cultures; 2 of the 6 had at least one ampicillin-resistant bacteria, and a third had 2 ampicillin-resistant bacteria . Eight patients who failed to improve with antimicrobial treatment had a second tympanocentesis performed or developed spontaneous drainage; on that follow-up culture 3 of 8 cultures had different microorganisms; and 5 of the 8 bacterial specimens were resistant to ampicillin or penicillin . Twenty-one percent of the S . pneumoniae strains recovered from the middle ear were resistant to penicillin . Sixty-two percent of the H . influenzae and 98% of the M . catarrhalis isolates were resistant to ampicillin . Overall bacteria resistant to penicillin or ampicillin were recovered in 54% of middle ear fluid from 46 patients who had received a beta-lactam antibiotic in the preceding month as well as in 57% of middle ear fluids from 105 patients who had not . The empiric use of amoxicillin for treatment of acute otitis media should be reexamined in our community particularly in those who appear ill, have a high fever or have severe unremitting otalgia. Ann Trop Paediatr, 1995 Dec, 15(4), 273 - 8 An ELISA assay for the rapid diagnosis of acute bacterial meningitis; Salih MA et al.; A recently introduced ELISA test (EIA-Test) was used for the detection of the common organisms that cause acute bacterial meningitis (ABM) . The test--which detects antigens of Haemophilus influenzae type b, Neisseria meningitidis (serogroups A, B, C) and Streptococcus pneumoniae (25 selected types)--is performed in less than 1 hour and read by the naked eye . Cerebrospinal fluid (CSF) specimens of 125 Sudanese children (1 month-14 years) admitted with a provisional diagnosis of meningitis/meningoencephalitis were prospectively examined with the EIA-Test, direct microscopy (DM) using Gram stain and culture . Of the 56 positive CSF specimens, the EIA-Test had the highest yield (100%), followed by culture (33.9%) and DM (30.4%) . Nineteen (33.9%) of the positive samples came from children who had been treated with antibiotics prior to admission, and all were positive in the EIA-Test . EIA-Test was also the only positive test in 31 (55.4%) cases . From the present study, and similar to previous observations, the EIA-Test seems to have valuable potential for the rapid diagnosis of ABM and is of particular help in patients who have been pre-treated with antibiotics. Eur J Clin Microbiol Infect Dis, 1995 Dec, 14(12), 1090 - 4 An approach to the identification of the pathogens of bacterial meningitis by the polymerase chain reaction; Hall LM et al.; A combination of universal and species-specific primers was used to detect and differentiate by nested polymerase chain reaction (PCR) the four species most commonly causing bacterial meningitis . Primers recognising conserved sequences in the 16S and 23S ribosomal RNA genes were employed to amplify the 16S-23S spacer region from Neisseria meningitidis, Haemophilus influenzae (type b), Streptococcus pneumoniae, and Streptococcus agalactiae (group B streptococcus) . The sequence of the most abundant spacer product was determined in each case and used to deduce species-specific primers . A nested PCR using universal primers in the first round and a species-specific primer in the second were able to detect and distinguish between the four common pathogens, in the presence of human DNA . Streptococcus pneumoniae DNA was detected in the cerebrospinal fluid of a meningitis patient with negative culture and Gram-stain results. Eur J Clin Microbiol Infect Dis, 1995 Dec, 14(12), 1088 - 90 Bacteremia and chorioamnionitis due to cryptic genospecies of Haemophilus Influenzae biotype I; Gill MV et al.; Nontypable strains of Haemophilus influenzae are well-known causes of maternal and neonatal infections . Using DNA-DNA hybridization techniques, some of these strains have been shown to belong to a cryptic genospecies of Haemophilus, which is distantly related to Haemophilus influenzae and Haemophilus hemolyticus . This report describes the first case of sepsis and chorioamnionitis due to Haemophilus influenzae biotype I, which was identified using the RapIDNH system and then confirmed by multilocus enzyme electrophoresis to belong to this cryptic genospecies of Haemophilus . The electromorph type 92 of the isolate was consistent with that of biotype I of the cryptic genospecies. Onderstepoort J Vet Res, 1995 Dec, 62(4), 261 - 70 Effects of transformation on the hemagglutinins of Haemophilus paragallinarum; Bragg RR et al.; Strain 0083 and two field isolates of H . paragallinarum were previously converted into NAD-independent organisms by the use of crude DNA extractions from naturally occurring NAD-independent H . paragallinarum isolates . Two of these transformed isolates {0083(T) and A745(T)} were used as DNA donors in another transformation experiment in which another field isolate (M85) was used as the DNA recipient . Transformation was confirmed by lack of NAD requirement for growth, by carbohydrate fermentation patterns and by a comparison of the monoclonal antibody patterns of the isolates before and after transformation . Previously, antigenic differences were observed when DNA from an NAD-independent isolate was introduced into strain 0083 . Antigenic differences were also seen in the transformed M85 organisms prepared in this work, and these differences were dependent on the antigenic patterns of the DNA donors . It was established by haemagglutination (HA) and haemagglutination inhibition (HI) that the hemagglutinins of 0083, A745/92 and M85 were not affected by transformation . The use of strains transformed to NAD independence for vaccine production appears to be a valid approach, as the transformation appears not to affect the hemagglutinins of the transformed organisms The major advantage would be the alleviation of the requirement for chicken serum or NAD in the bacterial growth medium used for infectious-coryza-vaccine production. J Chemother, 1995 Dec, 7(6), 530 - 4 Bactericidal kinetics and postantibiotic effect of sparfloxacin against selected species of respiratory pathogens; Speciale A et al.; We determined the bactericidal kinetics and postantibiotic effect (PAE) of sparfloxacin against Haemophilus influenzae, Moraxella (Branhamella) catarrhalis, Klebsiella pneumonia, Streptococcus pneumoniae, and Staphylococcus aureus . Time-kill studies were performed by using 1 x and 4 x the minimum inhibitory concentrations (MICs) of sparfloxacin, ciprofloxacin, co-trimoxazole, amoxicillin/clavulanic acid and erythromycin (inoculum 10(5) and 10(7) CFU/ml) . The PAE was induced by exposing the strains to 1xMIC and 4xMIC of sparfloxacin and ciprofloxacin for 1 h . Sparfloxacin was the most bactericidal of all the antibiotics tested, being active against Gram-positive and Gram-negative isolates with a 99.9% reduction within 3 to 6 h of exposure, depending upon strain, inoculum and concentration . The PAE of sparfloxacin against all species tested ranged from 1.1 to 2.6 hours; the most notable PAE occurring with M . catarrhalis. Eur Respir J, 1995 Dec, 8(12), 1999 - 2007 Treatment of community-acquired pneumonia: a randomized comparison of sparfloxacin, amoxycillin-clavulanic acid and erythromycin; Lode H et al.; The treatment of community-acquired pneumonia is empirical in most cases and must cover a wide range of potential pathogens, such as Streptococcus pneumoniae, including penicillin-resistant strains, Haemophilus influenzae and intracellular microorganisms . The objective of this double-blind, randomized, parallel group study was to compare the efficacy and safety of sparfloxacin (400 mg loading dose, followed by 200 mg o.d.) with that of oral amoxycillin-clavulanic acid (500/125 mg t.i.d.) or oral erythromycin (1 g b.i.d.), during 7-14 days in 808 patients with confirmed community-acquired pneumonia . The overall success rates for sparfloxacin (87%), amoxycillin-clavulanic acid (80%) and erythromycin (85%) were similar in evaluable patients, and the equivalence hypothesis used for the statistical analysis showed at least an equivalent efficacy for the three antibiotics tested . The analysis of microbiologically documented infections (40% of the patients) showed that overall success rates were similar for S . pneumoniae and H . influenzae infections . Treatment withdrawal was necessary in 3.5, 2.5 and 7.7% of the patients treated with sparfloxacin, amoxycillin-clavulanic acid and erythromycin, respectively . This study indicates that sparfloxacin was at least as effective as amoxycillin-clavulanic acid or erythromycin in the treatment of mild-to-moderate community-acquired pneumonia and that the adverse effects were similar in the three groups. Jpn J Antibiot, 1995 Dec, 48(12), 1920 - 34 {Antimicrobial activities of clavulanic acid/amoxicillin against freshly isolated clinical strains from outpatients}; Koguchi M et al.; In order to investigate antimicrobial activities of clavulanic acid/amoxicillin (CVA/AMPC) against freshly isolated clinical strains obtained in 1995, beta-lactamase activities and minimum inhibitory concentration (MICs) were determined including those of the control drugs . The results are summarized as follows; 1 . Detection frequencies of beta-lactamase producing strains were as follows: methicillin-susceptible Staphylococcus aureus subsp . aureus (MSSA, 90.0%), Haemophilus influenzae (22.0%), Moraxella subgenus Branhamella catarrhalis (100.0%), Escherichia coli (100.0%), Klebsiella pneumoniae subsp . pneumoniae (100.0%) and Neisseria gonorrhoeae (14.0%) . It appeared that beta-lactamases produced by these strains were mostly penicillinase or enzyme of similar that . 2 . Antimicrobial activities of CVA/AMPC against beta-lactamase producing strains were stronger than those of AMPC, and MIC90 of CVA/AMPC against benzylpenicillin (PCG)-insensitive or resistant Streptococcus pneumoniae was lower than those of sultamicillin, cefaclor and cefpodoxime . 3 . CVA showed strong beta-lactamase inhibitory effect against M.(B.) catarrhalis of direct and indirect pathogenicity . We can expect CVA/AMPC to negate or decrease the influence of indirect pathogenicity. J Clin Microbiol, 1995 Dec, 33(12), 3257 - 9 Enhanced recovery of Haemophilus ducreyi from clinical specimens by incubation at 33 versus 35 degrees C; Schmid GP et al.; Isolation rates of Haemophilus ducreyi from cases of chancroid are low . Experts recommend that isolation media be incubated at 33 to 35 degrees C, but the possible effect of this temperature range on the recovery of H . ducreyi has not been evaluated . We inoculated two sets of agar plates with material from genital ulcers and incubated one set at 33 degrees C and one at 35 degrees C; incubation at 33 degrees C identified 21% more cases than did incubation at 35 degrees C (109 versus 85 cases, respectively, of the 116 cases from which an isolation was made; P < 0.01). Thorax, 1995 Dec, 50(12), 1240 - 5 Pulmonary complications of HIV disease: 10 year retrospective evaluation of yields from bronchoalveolar lavage, 1983-93; Taylor IK et al.; BACKGROUND--Pulmonary disease is a major contributor to morbidity and mortality in patients with HIV infection and AIDS . The aim of this study was to describe bronchoscopic findings and the spectrum of pulmonary pathogens in HIV seropositive patients undergoing investigation of respiratory disease over a 10 year period in a major UK referral centre . METHODS--Recruitment was procedure based with data being captured when bronchoscopy was clinically indicated . Data were evaluated from 580 HIV seropositive patients (559 men, age 13-65 years) over a 10 year period from June 1983 to March 1993 . RESULTS--A total of 947 bronchoscopies was performed . The most frequent pulmonary pathogen isolated from bronchoalveolar lavage (BAL) fluid in 44% of all bronchoscopies was Pneumocystis carinii . Of all patients studied, 324 (55%) had at least one cytologically confirmed episode of P carinii pneumonia; this was AIDS defining in 219 (38%) of patients who underwent bronchoscopy . Between 1987 and 1993 the overall diagnostic yield from BAL fluid was 76%; 25% of all bronchoscopies yielded positive microbiological results, the most frequent isolates being Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas spp, and Haemophilus influenzae . Mycobacteria were identified in 8% of patients; M tuberculosis was the most common being identified in 3% of lavage samples and in 4% of patients . No drug-resistant M tuberculosis was found . Viral isolates (mainly cytomegalovirus) were identified in up to 31% of BAL fluid samples . Endobronchial Kaposi's sarcoma was seen in 15% of patients at bronchoscopy . CONCLUSIONS--Of the 1956 newly diagnosed HIV seropositive patients receiving clinical care at St Mary's Hospital over this period, approximately 30% underwent bronchoscopy . Diagnostic rates for P carinii pneumonia, endobronchial Kaposi's sarcoma, and bacterial and mycobacterial infection have remained largely constant since 1989 . Bronchoalveolar lavage produces high diagnostic yields generally, and P carinii pneumonia remains a common cause of pulmonary disease in these patients. J Bacteriol, 1995 Dec, 177(24), 7265 - 70 Characterization of Actinobacillus pleuropneumoniae riboflavin biosynthesis genes; Fuller TE et al.; In this paper, we report the identification, cloning, and complete nucleotide sequence of four genes from Actinobacillus pleuropneumoniae that are involved in riboflavin biosynthesis . The cloned genes can specify production of large amounts of riboflavin in Escherichia coli, can complement several defined genetic mutations in riboflavin biosynthesis in E . coli, and are homologous to riboflavin biosynthetic genes from E . coli, Haemophilus influenzae, and Bacillus subtilis . The genes have been designated A . pleuropneumoniae ribGBAH because of their similarity in both sequence and arrangement to the B . subtilis ribGBAH operon. J Bacteriol, 1995 Dec, 177(23), 6820 - 4 Structure-function relationship of bacterial prolipoprotein diacylglyceryl transferase: functionally significant conserved regions; Qi HY et al.; The structure-function relationship of bacterial prolipoprotein diacylgyceryl transferase (LGT) Has been investigated by a comparison of the primary structures of this enzyme in phylogenetically distant bacterial species, analysis of the sequences of mutant enzymes, and specific chemical modification of the Escherichia coli enzyme . A clone containing the gene for LGT, lgt, of the gram-positive species Staphylococcus aureus was isolated by complementation of the temperature-sensitive lgt mutant of E . coli (strain SK634) defective in LGT activity . In vivo and in vitro assays for prolipoprotein diacylglyceryl modification activity indicated that the complementing clone restored the prolipoprotein modification activity in the mutant strain . Sequence determination of the insert DNA revealed an open reading frame of 837 bp encoding a protein of 279 amino acids with a calculated molecular mass of 31.6 kDa . S . aureus LGT showed 24% identity and 47% similarity with E . coli, Salmonella typhimurium, and Haemophilus influenzae LGT.S . aureus LGT, while 12 amino acids shorter than the E . coli enzyme, had a hydropathic profile and a predicted pI (10.4) similar to those of the E . coli enzyme . Multiple sequence alignment among E . coli, S . typhimurium, H . influenzae, and S . aureus LGT proteins revealed regions of highly conserved amino acid sequences throughout the molecule . Three independent lgt mutant alleles from E . coli SK634, SK635, and SK636 and one lgt allele from S . typhimurium SE5221, all defective in LGT activity at the nonpermissive temperature, were cloned by PCR and sequenced . The mutant alleles were found to contain a single base alteration resulting in the substitution of a conserved amino acid . The longest set of identical amino acids without any gap was H-103-GGLIG-108 in LGT from these four microorganisms . In E . coli lgt mutant SK634, Gly-104 in this region was mutated to Ser, and the mutant organism was temperature sensitive in growth and exhibited low LGT activity in vitro . Diethylpyrocarbonate inactivated the E . coli LGT with a second-order rate constant of 18.6 M-1S-1, and the inactivation of LGT activity was reversed by hydroxylamine at pH 7 . The inactivation kinetics were consistent with the modification of a single residue, His or Tyr, essential for LGT activity. J Bacteriol, 1995 Dec, 177(23), 6745 - 50 Expression and characterization of the ponA (ORF I) gene of Haemophilus influenzae: functional complementation in a heterologous system; Sharma UK et al.; The coding sequence of the Haemophilus influenzae ORF I gene was amplified by PCR and cloned into different Escherichia coli expression vectors . The ORF I-encoded protein was approximately 90 kDa and bound 3H-benzyl-penicillin and 125I-cephradine . This high-molecular-weight penicillin-binding protein (PBP) was also shown to possess transglycosylase activity, indicating that the ORF I product is a bifunctional PBP . The ORF I protein was capable of maintaining the viability of E . coli delta ponA ponB::spcr cells in transcomplementation experiments, establishing the functional relevance of the significant amino acid homology seen between E . coli PBP 1A and 1B and the H . influenzae ORF I product . In addition, the physiological functioning of the H . influenzae ORF I (PBP 1A) product in a heterologous species established the ability of the enzyme not only to recognize the E . coli substrate but also to interact with heterologous cell division proteins . The affinity of the ORF I product for 3H-benzylpenicillin and 125I-cephradine, the MIC of beta-lactams for E . coli delta ponA ponB::spcr expressing the ORF I gene, and the amino acid alignment of the PBP 1 family of high-molecular-weight PBPs group the ORF I protein into the PBP 1A family of high-molecular-weight PBPs. Infect Immun, 1995 Dec, 63(12), 4883 - 9 Contribution of the major and minor subunits to fimbria-mediated adherence of Haemophilus influenzae to human epithelial cells and erythrocytes; van Ham SM et al.; Fimbriae are colonization factors of the human pathogen Haemophilus influenzae in that they mediate bacterial adherence to human eukaryotic cells . The contribution of the major (HifA) and putative minor (HifD and HifE) subunits of H . influenzae fimbriae to fimbria-specific adherence was studied by using mutants that were inactivated in distinct fimbrial genes . Both the major and minor subunits were required for adherence of H . influenzae to oropharyngeal epithelial cells and human erythrocytes carrying the AnWj antigen . Cloning of defined H . influenzae fimbrial genes in an Escherichia coli strain with type 1 fimbriae yielded recombinants expressing high amounts of HifA-containing H . influenzae fimbriae either with or without coexpression of both H . influenzae minor subunits . Both clones exhibited the specific adherence properties of H . influenzae fimbriae, implying that the minor H . influenzae subunits are dispensable for adherence and that the adhesive domain resides in the major subunit, HifA . In H . influenzae itself, the minor subunits probably affect adherence by raising the number of fimbriae above the minimal level required to establish adherence. Infect Immun, 1995 Dec, 63(12), 4729 - 37 Paracytosis of Haemophilus influenzae through cell layers of NCI-H292 lung epithelial cells; van Schilfgaarde M et al.; Haemophilus influenzae penetrates the respiratory epithelium during carriage and invasive disease, including respiratory tract infections . We developed an in vitro model system consisting of lung epithelial NCI-H292 cells on permeable supports to study the passage of H . influenzae through lung epithelial cell layers . The NCI-H292 cells formed tight layers with a Ca(2+)-dependent transepithelial resistance of around 40 omega.cm2 . H . influenzae passed through the cell layers without affecting the viability of the cells and {3H}inulin penetration . The passage time was independent of the inoculum of H . influenzae in the apical compartment and was not influenced by the presence of capsule or fimbriae on H . influenzae or by the ability of the bacteria to adhere to the epithelial cells . However, highly adherent strains showed greater paracytosis . Different strains passed through the cell layer independently . The passage time was shorter for rapidly growing strains than for slowly growing strains (10 to 18 h and 30 h, respectively) . Microscopic examination revealed the presence of clusters of H . influenzae bacteria between the epithelial cells, indicating that bacterial passage was due to paracytosis . After the addition of chloramphenicol, no bacteria were cultured from the basolateral side, and no bacterial clusters between the epithelial cells were seen, suggesting that de novo bacterial protein synthesis was needed for the bacteria to reach the intercellular space . We conclude that H . influenzae passes through viable cell layers of the human lung epithelial cell line NCI-H292 by paracytosis, requiring bacterial protein synthesis. Chest, 1995 Dec, 108(6), 1640 - 7 Acute epiglottitis . An 18-year experience in Rhode Island; Mayo-Smith MF et al.; OBJECTIVE: To assess the incidence, clinical characteristics, management, and outcome of epiglottitis in a defined population over an 18-year period . DESIGN: Case series . SETTING: The state of Rhode Island, 1975 through 1992 . PATIENTS OR OTHER PARTICIPANTS: Cases who met predetermined criteria for acute epiglottitis identified from hospital discharges and the State Medical Examiner's log of prehospitalization deaths . MAIN OUTCOME MEASURES: Incidence by year and age, clinical presentation, results of diagnostic evaluations, management, and outcome . RESULTS: Four hundred seven cases were identified, 134 in children and 273 in adults . Incidence in children dropped from 38 cases in the first 3 years of the study to 1 case in the last 3 years (p < 0.001) . Adult cases increased from 17 in the first 3 years to 69 in the last 3 years (p < 0.001) . Seventy-nine percent of adults and 32% of children were treated without an artificial airway . Factors associated with airway obstruction included symptomatic respiratory difficulty, stridor, drooling, shorter duration of symptoms, enlarged epiglottis on radiograph, and Haemophilus influenzae bacteremia (p < 0.001 for each) . Twelve patients died (3 children and 9 adults), with all cases of fatal respiratory obstruction occurring within 12 h of presentation . CONCLUSIONS: There have been significant changes in the clinical epidemiology of epiglottitis, which now occurs almost exclusively in adults, often with less severe symptoms and a lower incidence of H influenzae infection . While careful observation is indicated for all patients, the data suggest that those with certain clinical characteristics can be treated safely without an immediate artificial airway. Ann Intern Med, 1995 Dec 1, 123(11), 828 - 34 Antibody responses to polysaccharide and polysaccharide-conjugate vaccines after treatment of Hodgkin disease; Molrine DC et al.; OBJECTIVE: To compare the immunogenicity of polysaccharide-conjugate vaccines with that of polysaccharide vaccines in patients previously treated for Hodgkin disease . DESIGN: All patients were immunized with Haemophilus influenzae type b (HIB)-conjugate and 4-valent meningococcal polysaccharide vaccines . Subgroups of patients were randomly assigned to receive either 23-valent pneumococcal polysaccharide vaccine or a 7-valent pneumococcal-conjugate vaccine that links seven pneumococcal serotypes to the outer membrane protein complex of Neisseria meningitidis . PATIENTS: 144 patients who had completed treatment for Hodgkin disease, which had been diagnosed at least 2 years before the study . MEASUREMENTS: Antigen-specific antibody concentrations before and 3 to 6 weeks after immunization; number of persons who achieved anti-HIB antibody concentrations considered to be in the protective range . RESULTS: The geometric mean anti-HIB antibody concentration increased from 1.79 micrograms/mL before immunization to 54.1 micrograms/mL after; the percentage of persons with antibody concentrations in the protective range increased from 62% before immunization to 99% after . Patients immunized with 23-valent pneumococcal vaccine had a geometric mean pneumococcal antibody concentration after immunization (9.15 micrograms/mL) that was similar to that of healthy controls (10.0 micrograms/mL) for the seven serotypes measured . In contrast, patients who received 7-valent pneumococcal-conjugate vaccine had a significantly lower mean response compared with patients who received 23-valent; their geometric mean antibody concentration after immunization was 4.95 micrograms/mL (P = 0.005) . CONCLUSION: A single dose of HIB-conjugate vaccine was immunogenic in patients who had completed treatment for Hodgkin disease diagnosed at least 2 years before immunization . In addition, responses to the 23-valent pneumococcal and 4-valent meningococcal vaccines were equivalent to those seen in healthy controls . Finally, patients had a significantly lower response to a single dose of 7-valent pneumococcal-conjugate vaccine than to 23-valent vaccine. Commun Dis Rep CDR Rev, 1995 Nov 10, 5(12), R173 - 7 Management of asplenic patients in South Buckinghamshire: an audit of local practice; MacInnes J et al.; People without spleens have an increased risk of pneumococcal and other infections . Immunisation is advised for this group of patients, but the role of prophylactic antibiotics remains unresolved . Since 1992, general practitioners in South Buckinghamshire have been encouraged to immunise all asplenic patients against infections with Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae type b (Hib) . In addition, an 'alert' card, similar in principle to a medical warning bracelet, has been produced for general practitioners to issue to asplenic patients . General practitioners' clinical records of 293 asplenic patients were subsequently examined to evaluate this programme and assess the use of prophylactic antibiotics . Uptakes of 91%, 80%, and 79% were achieved for vaccines against pneumococcal, meningococcal, and Hib infections, respectively . Twenty-three per cent of patients had been advised immediately after splenectomy to take prophylactic antibiotics . Prophylaxis was advised for different periods of time, particularly in children . Thirty-four different antibiotic regimens had been recommended for adults . Clinical records suggested that 9% of patients were taking antibiotic prophylaxis at the time of the analysis . 'Alert' cards had been distributed to 88% of patients who were eligible . It is likely that most districts within the United Kingdom could set up similar immunisation and 'alert' card programmes . The wide variation in recommendations for antibiotic prophylaxis highlights the need for further research and the development of national guidelines. J Biol Chem, 1995 Nov 10, 270(45), 27151 - 9 Mutation of the htrB locus of Haemophilus influenzae nontypable strain 2019 is associated with modifications of lipid A and phosphorylation of the lipo-oligosaccharide; Lee NG et al.; The HtrB protein was first identified in Escherichia coli as a protein required for cell viability at high temperature, but its expression was not regulated by temperature . We isolated an htrB homologue from non-typable Haemophilus influenzae strain (NTHi) 2019, which was able to functionally complement the E . coli htrB mutation . The promoter for the NTHi 2019 htrB gene overlaps the promoter for the rfaE gene, and the two genes are divergently transcribed . The deduced amino acid sequence of NTHi 2019 HtrB had 56% homology to E . coli HtrB . In vitro transcription-translation analysis confirmed production of a protein with an apparent molecular mass of 32-33 kDa . Primer extension analysis revealed that htrB was transcribed from a sigma 70-dependent consensus promoter and its expression was not affected by temperature . The expression of htrB and rfaE was 2.5-4 times higher in the NTHi htrB mutant B29 than in the parental strain . In order to study the function of the HtrB protein in Haemophilus, we generated two isogenic htrB mutants by shuttle mutagenesis using a mini-Tn3 . The htrB mutants initially showed temperature sensitivity, but they lost the sensitivity after a few passages at 30 degrees C and were able to grow at 37 degrees C . They also showed hypersensitivity to deoxycholate and kanamycin, which persisted on passage . SDS-polyacrylamide gel electrophoresis analysis revealed that the lipo-oligosaccharide (LOS) isolated from these mutants migrated faster than the wild type LOS and its color changed from black to brown as has been described for E . coli htrB mutants . Immunoblotting analysis also showed that the LOS from the htrB mutants lost reactivity to a monoclonal antibody, 6E4, which binds to the wild type NTHi 2019 LOS . Electrospray ionization-mass spectrometry analysis of the O-deacylated LOS oligosaccharide indicated a modification of the core structure characterized in part by a net loss in phosphoethanolamine . Mass spectrometric analysis of the lipid A of the htrB mutant indicated a loss of one or both myristic acid substitutions . These data suggest that HtrB is a multifunctional protein and may play a controlling role in regulating cell responses to various environmental changes. Qual Health Care, 1995 Dec, 4(4), 269 - 72 Managing meningitis in children: audit of notifications, rifampicin chemoprophylaxis, and audiological referrals; Shields MD et al.; Important aspects of the management of meningitis in children include notification to local officers for control of communicable diseases; chemoprophylaxis for index cases and close contacts in cases of meningococcal or Haemophilus influenzae meningitis; and a formal hearing assessment for all survivors . A retrospective audit of these aspects of management was carried out for children admitted with meningitis in 12 months from 1 September 1990 to 31 August 1991 at the Royal Belfast Hospital for Sick Children . Only 20 of 36(56%) cases were notified by medical staff . Chemoprophylaxis was arranged for all close family contacts but to only five of the 23(22%) index cases for whom it was indicated . Appointments for audiological testing were arranged for only 19 of the 32(59%) survivors . Subsequently all doctors, including each intake of junior doctors, were given written information on the importance of notification and locally agreed guidelines for chemoprophylaxis and hearing assessments for survivors before discharge . Guidelines were also displayed prominently in each ward . A repeat audit from January 1992 to December 1992 showed significant improvement in these aspects of care . Twenty eight of 32 cases (88%) were notified, chemoprophylaxis was given to 20 of 22(91%) index cases for whom it was indicated, and 25 of 29(86%) survivors had hearing assessments arranged before discharge . Correct management of some aspects of care cannot be assumed, even if statutory (notification), nationally agreed (chemoprophylaxis), or generally agreed good practice (hearing assessments) . These aspects of care improved after the first audit but the authors conclude that the notification rate remains below 100% and a repeat audit is necessary. Diagn Microbiol Infect Dis, 1995 Nov, 23(3), 89 - 98 Laboratory diagnosis of chancroid using species-specific primers from Haemophilus ducreyi groEL and the polymerase chain reaction; Parsons LM et al.; To enhance laboratory identification of Haemophilus ducreyi, the causative agent of the genital ulcer disease chancroid, a polymerase chain reaction (PCR) assay was developed using target DNA sequences from the essential H . ducreyi gene, groEL . Positive reactions were obtained in this PCR assay with 139 isolates of H . ducreyi from patients in worldwide locations from the 1940s to the 1990s . In contrast, 24 other bacterial species were negative . When genital ulcer specimens from 162 African patients with clinically diagnosed chancroid were evaluated, 66 were culture positive . The sensitivity of PCR as compared with culture was 89% (59 of 66), and specificity was 79% (76 of 96) . However, representative samples of the 20 culture-negative, PCR-positive specimens were confirmed as positive by a second PCR assay using different H . ducreyi-specific primers . Thus, combined results of culture and PCR detected H . ducreyi in 86 specimens, with resolved sensitivities of 92% (79 of 86) for PCR, and 77% (66 of 86) for culture . These results suggest that PCR assays for H . ducreyi have great potential for augmenting or replacing problematic cultural techniques. Mol Microbiol, 1995 Nov, 18(4), 729 - 40 Molecular analysis of a locus for the biosynthesis and phase-variable expression of the lacto-N-neotetraose terminal lipopolysaccharide structure in Neisseria meningitidis; Jennings MP et al.; Lipopolysaccharide (LPS) is a major determinant of Neisseria, meningitidis virulence . A key feature of meningococcal LPS is the phase-variable expression of terminal structures which are proposed to have disparate roles in pathogenesis . In order to identify the biosynthetic genes for terminal LPS structures and the control mechanisms for their phase-variable expression, the lic2A gene, which is involved in LPS biosynthesis in Haemophilus influenzae, was used as a hybridization probe to identify a homologous gene in N . meningitidis strain MC58 . The homologous region of DNA was cloned and nucleotide sequence analysis revealed three open reading frames (ORFs), two of which were homologous to the H . influenzae lic2A gene . All three ORFs were mutagenized by the insertion of antibiotic-resistance cassettes and the LPS from these mutant strains was analysed to determine if the genes had a role in LPS biosynthesis . Immunological and tricine-SDS-PAGE analysis of LPS from the mutant strains indicated that all three genes were probably transferases in the biosynthesis of the terminal lacto-N-neotetraose structure of meningococcal LPS . The first ORF of the locus contains a homopolymeric tract of 14 guanosine residues within the 5'-end of the coding sequence . As the lacto-N-neotetraose structure in meningococcal LPS is subject to phase-variable expression, colonies that no longer expressed the terminal structure, as determined by monoclonal antibody binding, were isolated . Analysis of an 'off' phase variant revealed a change in the number of guanosine residues resulting in a frameshift mutation, indicating that a slipped-strand mispairing mechanism, operating in the first ORF, controls the phase-variable expression of lacto-N-neotetraose. Kansenshogaku Zasshi, 1995 Nov, 69(11), 1227 - 34 {Usefulness of endotoxin-specific limulus test for the measurement of endotoxin in cerebrospinal fluid in diagnosis of bacterial meningitis}; Ichinohe S et al.; Using a new endotoxin-specific chromogneic limulus assay (Endoscopy test), endotoxin concentrations were measured in 93 specimens of cerebrospinal fluid (CSF) from 66 pediatric patients . Eighteen patients were diagnosed as having menigitios . Of these, 6 cases (group A) with gram-negative meningitis proven by culture had high CSF endotoxin concentrations of 115.3, (82-133) (median, range) pg/ml . Ten cases (group B) with gram-positive or aseptic meningitis had endotoxin concentrations of 2.15 (0.1-3.6) pg ml . Other 2 cases with bacterial meningitis (group C), in whom no pathogen was detected, had CSF endotoxin concentrations of more than 100 pg/ml . Four cases with encephalitis (group D) and 45 cases with non-meningitis or non- encephalitis (group E), had CSF endotoxin concentrations of less than 5 pg/ml . Despite a negative culture after antibiotic treatment in group A patients, endotoxin was cleared slowly from the CSF . A clearing of endotoxin from CSF was followed by alleviation of fever with a more gradual decline in CRP values . In 2 cases of group C, the negative bacterial culture appeared to be attributable to the previous treatment with antibiotics . However, these patients had high CSF endotoxin levels, indicating gram negative bacterial meningitis . In 17 CSF specimens from 5 patients of group A, in whom Haemophilus influenzae was detected on admission, an additional a latex agglutination test for the detection of H . influenzae polysaccharide antigen was performed . Only 3 specimens from 3 patients with CSF endotoxin concentrations of more than 80 pg/ml had a positive agglutination test . These results suggest that quantitation of endotoxin concentrations is useful for the diagnosis of gram-negative meningitis . And also, the clearance of endotoxin from CSF during treatment appears to be useful in determining the timing of when antibiotic should be stopped. Infection, 1995 Nov-Dec, 23(6), 391 - 2 Haemophilus parainfluenzae: an unusual case of psoas abscess; Laing RB et al.; A case of Haemophilus parainfluenzae psoas abscess in a previously healthy 36-year-old man is reported here . The absence of any bowel pathology indicates that abscess formation occurred secondary to haematogenous spread of the organism. Eur J Clin Microbiol Infect Dis, 1995 Nov, 14(11), 935 - 48 Haemophilus influenzae: then and now; Jordens JZ et al.; Haemophilus influenzae has long been recognised as a major cause of serious infection and mortality in children less than 5 years old . Prior to the introduction of Haemophilus influenzae type b (Hib) immunisation, the incidence of a child suffering an invasive Haemophilus infection was 20-50/100,000 in industrialised countries and up to ten times higher in developing regions . The introduction of a Hib vaccine programme results in a rapid and dramatic decline in the incidence of Hib infection in the susceptible childhood population . For example, within two years of the introduction of routine Hib vaccination of infants in the UK, the risk of serious Hib infection had fallen from 1:600 to 1:30,000 by 5 years of age . Many other European countries have introduced, or are in the process of introducing, a routine Hib immunisation programme . Because the epidemiology of Haemophilus influenzae infection is changing so dramatically, it is opportune to review Haemophilus influenzae as it was perceived in the pre-vaccine era (the past) and during vaccine implementation (the present), and how its role may change in the post-vaccination era (the future) . This review will summarise the historical landmarks that have led to our present-day understanding of Haemophilus influenzae pathogenicity, the concerns about antibiotic resistance, the features of the host immune response to Haemophilus influenzae, and the introduction of the Hib vaccine . Furthermore, the possible importance of this organism in the future will be discussed. Australas J Dermatol, 1995 Nov, 36(4), 216 - 8 Paediatric vulvovaginitis; O'Brien TJ; Vulvovaginitis in the prepubertal child may present with itch, irritation, pain, dysuria or discharge as a result of poor hygiene, contact irritants, sexual abuse or skin disease involving the genital area . Common infectious causes include threadworms, group A beta-haemolytic streptococci and Haemophilus influenzae . A full history and examination are essential . Investigations may include swabs, midstream urine, biopsy, ultrasound and examination under anaesthesia. Clin Infect Dis, 1995 Nov, 21(5), 1322 - 4 The emergence of Haemophilus influenzae types e and f as significant pathogens; Waggoner-Fountain LA et al.; Non-type b encapsulated Haemophilus influenzae meningitis (two cases due to H . influenzae type e, two due to H . influenzae type f) was diagnosed in four children in a 6-month period at the University of Virginia . H . influenzae type b was the most common cause of bacterial meningitis in the United States before the introduction of an effective vaccine, whereas the other five encapsulated serotypes of H . influenzae rarely caused invasive disease . The clinical features of non-type b H . influenzae meningitis and the therapy for this infection are the same as those for type b H . influenzae disease . We report these four cases to document an increase in infection due to non-type b serotypes of H . influenzae, and we postulate that this change may result from the well-documented decrease in H . influenzae type b oropharyngeal carriage and disease that has occurred because of universal vaccination for H . influenzae type b. Clin Infect Dis, 1995 Nov, 21(5), 1193 - 6 Development of beta-lactamase-mediated resistance to penicillin in middle-ear isolates of Moraxella catarrhalis in Finnish children, 1978-1993; Nissinen A et al.; The frequency of beta-lactamase production was analyzed in a study of 1,452 strains of Moraxella catarrhalis and 2,738 strains of Haemophilus influenzae isolated from middle-ear fluid of children < 6 years of age at Tampere University Hospital in Tampere, Finland, between 1978 and 1993 . In addition, 401 isolates of M . catarrhalis from similar samples collected in different parts of Finland in 1988-1990 were tested for beta-lactamase production; minimal inhibitory concentrations of ampicillin, cefaclor, cephalothin, erythromycin, tetracycline, and trimethoprim-sulfamethoxazole for these strains were determined . These data were compared with figures for the annual consumption of beta-lactam antimicrobials in the community in 1978-1993 . A bimodal increase in the proportion of strains of M . catarrhalis producing beta-lactamase was detected: from 0 to 60% in 1978-1983 and from 60% to 80% in 1988-1990 . Concurrently with the second increase, the consumption of cephalosporins increased substantially in the community . The frequency of beta-lactamase-producing strains of H . influenzae did not increase between 1978 and 1993. Clin Infect Dis, 1995 Nov, 21(5), 1134 - 44 Spectrum of 2,836 cases of invasive bacterial or fungal infections in children: results of prospective nationwide five-year surveillance in Finland . Finnish Pediatric Invasive Infection Study Group; Saarinen M et al.; In a prospective nationwide laboratory-based surveillance study of all invasive bacterial and fungal infections among children < 16 years of age, 2,836 clinical cases were registered during the 5-year period 1985-1989 . Of these cases, 136 were polymicrobial . During the study period, nationwide administration of Haemophilus influenzae type b conjugate vaccine reduced the incidence rates of invasive infection caused by this organism . The most common clinical diagnosis (48% of cases) was bacteremia without an identified focus of infection . The age-specific annual incidence rates of all invasive infections in children < or = 15 years of age, in children < or = 4 years of age, in children < or = 1 year of age, and in children < or = 28 days of age were 55.8, 141.4, 272.7, and 2,749.0 cases/100,000 person-years, respectively . Thirty percent of the children in the study had an underlying condition predisposing to infection . The case-fatality rate was 4.1% for all cases of invasive infection. J Hosp Infect, 1995 Nov, 31(3), 219 - 24 Contamination of multi-use closed tracheal suction catheters: an in-vitro study; Webb CH et al.; The use of closed multi-use tracheal suction systems in ventilated patients is increasingly popular as it offers a number of potential physiological and microbiological advantages . As the use of these systems has been associated with an increased prevalence of tracheal colonization, effective self-cleaning mechanisms employed in these catheters are necessary . Using an in-vitro model we assessed the degree of catheter colonization in two multi-use catheter systems; Stericath and Trach-Care . Fifty of each catheter type were tested for colonization with five challenge organisms: Staphylococcus aureus (NCTC 6571); Pseudomonas aeruginosa (NCTC 10662); Haemophilus influenzae type b; S . epidermidis (slime producer); and S . epidermidis (slime non-producer) . All five challenge strains had colonized both catheter types after 24 h and all 100 catheter tips were colonized . The extent and density of colonization was variable and interspecies variation in the pattern of catheter colonization was observed . The mean colonized lengths of Stericath catheters was 10.9 +/- 1.4 cm and of Trach-Care 2.7 +/- 1.3 cm (P < 0.0001). Drugs, 1995 Nov, 50(5), 838 - 53 Bacterial meningitis . Practical guidelines for management; Rockowitz J et al.; The therapy of bacterial meningitis has evolved over the past century . Initially, antimeningococcal antiserum was used to treat patients with meningococcal meningitis . During the 1930s, sulphonamides were the first antibiotics used in the treatment of bacterial meningitis . The use of other antibiotics followed in later decades . Insights into the pathophysiology of meningitis have led to the use of prophylaxis against infection, as well as adjunctive therapy aimed at attenuating the harmful sequelae, should infection occur . This article outlines the basic principles important in the selection of appropriate antimicrobials . the emergence of resistant organisms, specifically Streptococcus pneumoniae and Haemophilus influenzae, has necessitated changes in previously effective antimicrobial regimens . The availability of third generation cephalosporins has increased the survival rate for meningitis caused by Gram-negative bacilli . Research into the use of adjunctive steroids has led to the recommendation that these agents be used in the paediatric population, which traditionally has had a high prevalence of H . influenzae meningitis . The high efficacy of H . influenzae type b conjugate vaccine and the observation that steroids, by decreasing inflammation, also decrease CNS penetration of some drugs, has led to reconsideration of routine steroid use . Effective chemoprophylactic regimens for contacts of patients with either H . influenzae or Neisseria meningitidis can diminish the spread of infection . Vaccination for both immunocompetent and immunodeficient patients protects against disease caused by some of the more common meningeal pathogens. Antimicrob Agents Chemother, 1995 Nov, 39(11), 2528 - 34 Effectiveness of short-course therapy (5 days) with cefuroxime axetil in treatment of secondary bacterial infections of acute bronchitis; Henry D et al.; Five hundred thirty-seven patients were enrolled in two independent, investigator-blinded, multicenter, randomized clinical trials comparing the clinical and bacteriologic efficacies and the safety of 5- or 10-day treatment with cefuroxime axetil with those of 10-day treatment with amoxicillin-clavulanate in the treatment of secondary bacterial infections of acute bronchitis . Patients received either 5 or 10 days of treatment (n = 177 in each group) with cefuroxime axetil at 250 mg twice daily or 10 days of treatment (n = 183) with amoxicillin-clavulanate at 500 mg three times daily . Patients in the cefuroxime axetil (5 days) group received placebo on days 6 to 10 . Bacteriologic assessments were based on sputum specimen cultures obtained preceding and, when possible, following treatment . Organisms were isolated from the pretreatment sputum specimens of 242 of 537 (45%) patients, with the primary pathogens being Haemophilus influenzae, Haemophilus parainfluenzae, Moraxella catarrhalis, Streptococcus pneumoniae, and Staphylococcus aureus (28, 25, 13, 9, and 8% of isolates, respectively) . Pathogens were eradicated or presumed to be eradicated in 87% (52 of 60), 91% (53 of 58), and 86% (60 of 70) of bacteriologically evaluable patients treated with cefuroxime axetil (5 days), cefuroxime axetil (10 days), and amoxicillin-clavulanate, respectively . A satisfactory clinical outcome (cure or improvement) was achieved in 82% (107 of 130), 86% (117 of 136), and 83% (130 of 157) of the clinically evaluable patients treated with cefuroxime axetil (5 days), cefuroxime axetil (10 days), and amoxicillin-clavulanate, respectively . Treatment with amoxicillin-clavulanate was associated with a significantly higher incidence of drug-related adverse events than was treatment with cefuroxime axetil for either 5 or 10 days (P = 0.001), primarily reflecting a higher incidence of drug-related gastrointestinal adverse events (37 versus 19 and 15%, respectively; P < 0.001), particularly diarrhea and nausea . These results indicate that treatment with cefuroxime axetil at 250 mg twice daily for 5 days is as effective as treatment for 10 days with either the same dose of cefuroxime axetil or amoxicillin-clavulanate at 500 mg three times daily in patients with acute bronchitis . In addition, treatment with cefuroxime axetil for either 5 or 10 days is associated with significantly fewer gastrointestinal adverse events, particularly diarrhea and nausea, than is 10-day treatment with amoxicillin-clavulanate. Antimicrob Agents Chemother, 1995 Nov, 39(11), 2516 - 9 Resistance to ceftriaxone and other beta-lactams in bacteria isolated in the community . The Vigil'Roc Study Group; Goldstein FW et al.; The incidence of bacterial species and their susceptibilities to ceftriaxone and other beta-lactams from patients with community-acquired infections were evaluated in a multicenter study over a 4-month period . A total of 5,768 bacterial isolates were classified according to whether the patient had been previously hospitalized or had received antibiotic treatment . The most relevant findings were the presence of 33.8% penicillin-resistant Streptococcus pneumoniae isolates, 25% beta-lactamase-producing Haemophilus influenzae isolates, and 36.4% amoxicillin-resistant Escherichia coli isolates . All of these bacteria were fully susceptible to ceftriaxone . Nosocomial multiply-resistant bacteria, and particularly methicillin-resistant S . aureus, were found, as expected, at a higher frequency among previously hospitalized patients . However, such bacteria may be present in the community; their incidence is high in particular clinical settings, and such bacteria should be considered when one is choosing a first-line therapy for the treatment of severe infections. Pediatr Infect Dis J, 1995 Nov, 14(11), 978 - 81 Postlicensure surveillance for Haemophilus influenzae type b invasive disease after use of Haemophilus influenzae type b oligosaccharide CRM197 conjugate vaccine in a large defined United States population: a four-year eight-month follow-up; Shinefield HR et al.; Haemophilus influenzae type b (Hib) oligosaccharide CRM197 (HbOC-HibTITER, Lederle-Praxis) conjugate vaccine was approved for use in infants in October, 1990, as a result of an evaluation of 61,080 infants in whom the vaccine was found to be safe and effective . This report details a 4-year 8-month period of surveillance after the licensure of this vaccine in the large defined United States population from which these children were drawn . The population analyzed consisted of approximately 240,000 children from 0 through 8 years of age in a total health plan population of 2,500.000 . During this period 16 cases of invasive Hib disease were identified . Nine had not received any type of Hib vaccine, 2 had received the unconjugated polysaccharide Hib vaccine and 4 had received the HbOC vaccine . Of this latter group one toddler had received a single dose of the vaccine at 18 months of age < 24 hours before becoming ill, two 7-month-old infants had received a single dose of the vaccine at 2 or 3 months of age and one 3-year 4-month-old child had received three doses of HbOC vaccine as an infant but had never received the recommended booster dose . The 16th case occurred on July 10, 1994, in a 7- year-old child who at 18 months of age had received a single dose of PRP-D (ProHIBIT; Connaught) conjugate Hib vaccine . The use of HbOC conjugate Hib vaccine in this population has coincided with the virtual disappearance of invasive Hib disease in children 0 through 8 years of age with no cases of invasive Hib disease occurring in the past 3 years and 4 months after the use of this vaccine . During this period no concomitant increase of H . influenzae disease with other types of H . influenzae organisms has been seen in the study population. Acta Paediatr, 1995 Nov, 84(11), 1262 - 7 Carriage of Haemophilus influenzae and Streptococcus pneumoniae in healthy Chinese and Vietnamese children in Hong Kong; Sung RY et al.; Nasopharyngeal carriage of Haemophilus influenzae and Streptococcus pneumoniae was studied in 621 healthy Chinese children and 300 healthy Vietnamese children aged from 2 months to 5 years in Hong Kong . The carriage rate of H . influenzae type b in Vietnamese children was 1.3% (CI 0.04-2.63); it was zero in Chinese . The carriage rate of non-typable H . influenzae was 5.8% (CI 1.4-7.6%) in Chinese and 65.4% (CI 58.9-69.8%) in Vietnamese . The carriage rates of S . pneumoniae were 10.8% (CI 8.3-13.2%) and 55.7% (CI 50.1-61.3%) in Chinese and Vietnamese children, respectively . Univariate and multivariate logistic regression analyses were performed to search for factors associated with differences in carriage rates of both H . influenzae and S . pneumoniae between Chinese and Vetnamese children . Although older age, smaller living area and parental smoking were associated with higher carriage rates, these could not explain the remarkably low carriage rates of both bacteria in Chinese children. Vaccine, 1995 Nov, 13(16), 1563 - 6 Economic aspects of a general vaccination against invasive disease caused by Haemophilus influenzae type b (Hib) via the experience of the Children's Hospital La Fe, Valencia, Spain; Asensi F et al.; With the aim of studying whether a general vaccination against invasive disease caused by Haemophilus influenzae type b (Hib) is economically profitable bearing in mind the efficacy and safety of the vaccine, its price and the global cost that this disease has in our area, a review is conducted of patients admitted due to invasive disease caused by Hib in the Children's Hospital La Fe, Valencia, born between 1984 and 1993 . They total 100, 63 who have meningitis . In the 81 cases (56 with meningitis) born between 1984 and 1990 (years that can be regarded as "closed" since all the patients were younger than 5 years of age) the total cost has been calculated for hospitalization, care during the acute phase, care for the sequelae (6 severe and 7 mild) and death (5 cases) . The mean annual cost of care can be calculated at 62 million pesetas, without making an economic valuation of the loss of life, and at 205 million pesetas taking this factor into account . The annual cost of vaccinating the 7000 babies under one year of age and falling within the Hospital's catchment area, on the basis of a vaccination pattern of three doses (at 2, 4 and 6 months) or four doses (at 2, 4, 6 and 15 months) would amount to 63 or 84 million pesetas, normal price to public (not covered by National Health Service), and 40 or 51 million pesetas if acquired by National Health Service . As a conclusion we can state that, even from the economic point of view, without quantifying the cost of the loss of life, a public general anti-Hib vaccination would be profitable in our area since it would mean an administration cost lower than that of the care required by patients . This is without taking into account the fact that emotional, family and social serious disturbances would also be avoided due to hospitalization, sequelae and deaths caused by a disease which is today perfectly preventable. Vaccine, 1995 Nov, 13(16), 1547 - 50 Antibody responses and symptoms after DTP and either tetanus or diphtheria Haemophilus influenzae type B conjugate vaccines given for primary immunisation by separate or mixed injection; Begg NT et al.; The safety and immunogenicity of two conjugate Haemophilus influenzae type B (Hib) vaccines administered either mixed with, or in separate limbs to, a whole-cell DTP vaccine, was compared in infants vaccinated at 2, 3 and 4 months of age . Antibody titres to purified polyribosylribitol phosphate, diphtheria, and to pertussis antigens between infants who received the Hib and DPT vaccines in separate limbs or in the same limbs were similar (P > 0.1) while antibody titres to tetanus toxoid were higher in the later group (P < 0.05) . This study demonstrated that both Hib vaccines can be mixed with whole-cell DTP vaccine without reducing immunogenicity of either vaccine or increasing the incidence of adverse reactions. J Clin Microbiol, 1995 Nov, 33(11), 3077 - 9 Evaluation of sampling sites for detection of upper respiratory tract carriage of Streptococcus pneumoniae and Haemophilus influenzae among healthy Filipino infants; Capeding MR et al.; Two sampling techniques, nasal swabbing and oropharyngeal swabbing, for detection of the upper respiratory tract carriage of Streptococcus pneumoniae and Haemophilus influenzae were studied prospectively with 296 healthy Filipino infants at various ages: 6 to 8, 10 to 12, 14 to 17, 18 to 22, 32 to 39, and 46 to 65 weeks . In all age groups S . pneumoniae was isolated significantly more often (P < 0.0001) from the nasal site than from the oropharyngeal site . H . influenzae was found equally often at both sites. J Clin Microbiol, 1995 Nov, 33(11), 3042 - 3 Haemophilus parainfluenzae biliary tract infection: rationale for an ascending route of infection from the gastrointestinal tract; Bottone EJ et al.; Haemophilus parainfluenzae was isolated from a bile specimen of a 56-year-old patient with elevated liver enzymes and chronic cholecystitis . Several factors contributing to nonbacteremic H . parainfluenzae infection of the biliary tract include (i) the not infrequent occurrence of H . parainfluenzae in stool specimens, (ii) the presence of an outer membrane protein similar to those found in enteric bacteria which facilitates adherence to the colonic mucosa, and (iii) an adequate supply of V factor (nicotinamide adenine dinucleotide) supplied from bile and the local flora, which is necessary for H . parainfluenzae's growth . Because H . parainfluenzae rarely occurs in the biliary tract, direct cultivation of bile specimens on chocolate agar seems unwarranted . However, bile specimens showing a gram-negative coccobacillus on a Gram-stained smear that fails to grow on routine media should be subcultured on chocolate agar. Am J Otolaryngol, 1995 Nov-Dec, 16(6), 383 - 90 Experimental recurrent otitis media induced by Haemophilus influenzae: protection and serum antibodies; Melhus A et al.; PURPOSE: To study whether acute otitis media caused by encapsulated or nontypeable Haemophilus influenzae confers cross-reactive protective immunity in an animal model system and to explore the possible involvement of various humoral specific antibodies in protection . MATERIALS AND METHODS: Rats were intrabullarly challenged with H influenzae type b and two different nontypeable H influenzae strains . One month after the initial infection, the animals were rechallenged ipsilaterally or contralaterally with either a homologous or heterologous strain, and the susceptibility to reinfection was investigated by otomicroscopy . RESULTS: The animals challenged and rechallenged with the type b strain were well-protected ipsilaterally and contralaterally, while the protection after homologous rechallenge with a nontypeable strain was partial in the ipsilateral ear and very poor in the contralateral ear . Middle ears previously infected with a nontypeable strain remained fully susceptible to infections with heterologous strains, but there was an indication of cross-protection in the animal groups where the first episode of acute otitis media was caused by type b and the second by a nontypeable strain . Using the Western blot technique and an enzyme linked immunosorbant assay, the serological response to different outer membrane proteins, especially protein D, of H influenzae during and after middle ear infection were investigated . The serological response from the type b infected animals were generally more distinct, while the antibody levels against protein D were lower in these groups compared with the groups infected with nontypeable strains . CONCLUSIONS: These data indicate that H influenzae type-b-induced experimental otitis media results in a better protection than a nontypeable-induced, and H influenzae b confers a cross protection. Protein Sci, 1995 Nov, 4(11), 2335 - 40 Modeling of the structure of the Haemophilus influenzae heme-binding protein suggests a mode of heme interaction; Dunten P et al.; The structure and function of the periplasmic heme-binding protein HbpA of Haemophilus influenzae were investigated . This protein is involved in the import of heme into the bacteria through the inner membrane, and thus is a key element of the organism's ability to survive in blood . A high degree of sequence similarity between HbpA and the dipeptide-binding protein of Escherichia coli is suggested to be the result of a functional relationship . An HbpA model built using the dipeptide-binding protein suggests a mode of heme binding that is distinct from those known in proteins of the human host . These results provide a starting point for rational drug design. APMIS, 1995 Nov, 103(11), 806 - 12 Haemophilus influenzae release histamine and enhance histamine release from human bronchoalveolar cells . Examination of patients with chronic bronchitis and controls; Clementsen P et al.; Haemophilus influenzae (H . influenzae), Streptococcus pneumoniae (S . pneumoniae) and Branhamella catarrhalis (B . catarrhalis) are often found in the lower respiratory tract of patients with chronic bronchitis . Earlier studies have shown that bacteria induce mediator release from human basophils and parenchymal lung mast cells . In this study the capability of bacteria to trigger or potentiate histamine release from superficially located mast cells in the airway epithelium was stu