Haemophilus

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 studied in cell suspensions obtained by bronchoalveolar lavage in patients with chronic bronchitis (CB) . In approximately half of the patients H . influenzae and Staphylococcus aureus (S . aureus) were found to trigger histamine release, whereas no response was obtained by S . pneumoniae or B . catarrhalis . The mediator release was caused by a non-IgE-dependent mechanism . At lower concentrations of H . influenzae causing no histamine release the bacterium was found to enhance IgE-mediated histamine release triggered by anti-IgE antibody . The synergy was more pronounced in patients with CB than in controls . Since H . influenzae is found in the lower respiratory tract of the patients but not in normal individuals, the infection here may via histamine release lead to harmful effects on the airways of importance for precipitation and exacerbation of chronic bronchitis.

Chemotherapy, 1995 Nov-Dec, 41(6), 437 - 47
Comparison of Wilkins-Chalgren medium supplemented with nicotinamide adenine dinucleotide and Haemophilus test medium for susceptibility testing of clinical Haemophilus isolates; Traub WH et al.; Haemophilus test medium (HTM) was compared with Wilkins-Chalgren agar (WCA; supplemented with 15 micrograms/ml nicotinamide adenine dinucleotide (NAD)) for antibiotic susceptibility testing of 74 clinical isolates of Haemophilus influenzae and Haemophilus parainfluenzae . The Bauer-Kirby agar disk diffusion method and the agar dilution procedure were the two tests employed . WCA + NAD and HTM yielded comparable results for the following antimicrobial drugs and drug combinations: ampicillin, amoxicillin plus clavulanic acid, ampicillin plus sulbactam, cefotaxime, cefuroxime, chloramphenicol, ciprofloxacin, and doxycycline . However, WCA + NAD slightly antagonized rifampin . Co-trimoxazole was significantly antagonized by WCA + NAD . WCA + NAD was much simpler with regard to in-house preparation, and WCA plates could be supplemented with NAD following pouring, solidification and storage of the plates simply by surface spreading the required amount of NAD with a glass spatula, thus adding considerably to laboratory flexibility . It is suggested that additional laboratories comparatively and critically examine WCA + NAD medium for antibiotic susceptibility testing of clinical Haemophilus isolates.

J Infect Dis, 1995 Nov, 172(5), 1388 - 92
Multiple Haemophilus influenzae strains and strain variants coexist in the respiratory tract of patients with cystic fibrosis; Moller LV et al.; To investigate the epidemiology of nontypeable Haemophilus influenzae in the respiratory tract of cystic fibrosis (CF) patients, H . influenzae isolates from sputum specimens of 40 CF patients were analyzed longitudinally for 2 years . The isolates were characterized by analysis of the major outer membrane protein (MOMP) patterns . MOMP variant H . influenzae strains were discriminated from distinct strains by randomly amplified polymorphic DNA analysis of genomic DNA . Multiple H . influenzae strains and MOMP variant strains were isolated from single sputum specimens of 29 patients . In 22 patients, a distinct H . influenzae strain persisted over time (median persistence, 8 months; range 2-24) . In general, the appearance of MOMP variant strains did not coincide with the occurrence of exacerbations.

J Infect Dis, 1995 Nov, 172(5), 1348 - 51
Influenza A virus--induced acute otitis media; Buchman CA et al.; To better understand the significance of viral upper respiratory tract infections in the pathogenesis of acute otitis media (OM), 27 adults underwent intranasal inoculation with influenza A virus . Monitoring consisted of antibody titer determination, tympanometry, and otoscopy . Microbiologic analysis consisted of cultures and polymerase chain reaction (PCR)-based detection for influenza A virus, Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis . All subjects became infected with the challenge virus . By day 4, 16 (59%) developed middle ear pressures of -100 mm H2O or below and 4 (25%) of them developed OM . One subject (4%) developed purulent OM requiring myringotomy for pain relief . Middle ear effusion cultures were negative . PCR analysis of that subject's middle ear effusion and nasal washes were positive for influenza A virus and S . pneumoniae . These findings support a causal role for viral upper respiratory tract infections in the pathogenesis of OM, possibly mediated by middle ear underpressures and viral and bacterial middle ear infection.

J Infect Dis, 1995 Nov, 172(5), 1330 - 5
The etiology of community-acquired pneumonia among hospitalized patients during a Chlamydia pneumoniae epidemic in Finland; Kauppinen MT et al.; The etiology of community-acquired pneumonia during a Chlamydia pneumoniae epidemic was studied among 125 hospitalized patients . Etiologic investigations included blood and sputum cultures, pneumococcal antigen detection, and serologic investigations for common respiratory viruses and for Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Mycoplasma pneumococcal antigen detection, and serologic investigations for common and for Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Mycoplasma pneumoniae, and Chlamydia species . A specific microbial agent was identified in 110 patients (88%) . Some 48% of the patients had mixed infections . S . pneumoniae was the most common pathogen (55%), followed by C . pneumoniae (43%) . C . pneumoniae was found both as a single etiologic agent and as a mixed infection, most often with S . pneumoniae . In conclusion, S . pneumoniae is the most common cause of community-acquired pneumonia, even during a C . pneumoniae epidemic . The organism, alone or with other pathogens (especially S . pneumoniae), may cause community-acquired pneumonia that requires hospital treatment.

J Infect Dis, 1995 Nov, 172(5), 1279 - 89
Complement-mediated bactericidal activity of human antibodies to poly alpha 2-->8 N-acetylneuraminic acid, the capsular polysaccharide of Neisseria meningitidis serogroup B; Mandrell RE et al.; Serum antibodies to Neisseria meningitidis group B (MenB) polysaccharide are reported not to elicit bacteriolysis in the presence of human complement . To reexamine this question, we evaluated the ability of two human IgM anti-MenB polysaccharide monoclonal antibodies (MAbs) and seven human MenB polysaccharide-reactive human IgM paraproteins to elicit bacteriolysis . In the presence of human complement, both MAbs and five of the seven paraproteins were bactericidal at antibody concentrations of 0.25-9.6 micrograms/mL (50% killing) . Activity of the respective antibodies was enhanced 200- to > 10,000-fold when rabbit complement was used instead of human complement . With rabbit complement, the bactericidal activity of human IgM polyclonal antibody or MAb to Haemophilus influenzae type b (Hib) polysaccharide but not human IgG polyclonal antibody or MAb to Hib polysaccharide was similarly augmented . Thus, for both MenB and Hib, IgM antipolysaccharide antibodies elicit complement-mediated bactericidal activity in the presence of human complement, and the use of rabbit complement yields spuriously high activity.

J Infect Dis, 1995 Nov, 172(5), 1268 - 72
Immunologic priming by one dose of Haemophilus influenzae type b conjugate vaccine in infancy; Kurikka S et al.; Immunogenicity of one dose of Haemophilus influenzae type b (Hib) conjugate vaccine in infancy and its ability to induce immunologic memory was studied in infants immunized at 4 and 14 months with either PRP-OMP (Hib polysaccharide conjugated with Neisseria meningitidis group B outer membrane protein complex) or PRP-T (Hib polysaccharide-tetanus toxoid conjugate) and compared with three doses of the same vaccines at 4, 6, and 14 months . Each group received diphtheriatetanus-pertusis vaccine at 3, 4, and 5 months of age . At 7 months of age, both vaccines were immunogenic after one dose, even though higher antibody concentrations were achieved after two doses . A booster dose given at 14 months resulted in a high antibody concentration and a strongly IgG-dominated isotype distribution, speaking for a secondary-type response in all groups, including those who had received only one dose in infancy . Subsequent persistence of antibodies suggestive of full protection for up to 36 months was similar in all groups.

Infect Immun, 1995 Nov, 63(11), 4433 - 8
The OmpU outer membrane protein, a potential adherence factor of Vibrio cholerae; Sperandio V et al.; Expression of the OmpU outer membrane protein of Vibrio cholerae is positively regulated by toxR, which also regulates critical virulence factors such as cholera toxin and the toxin-coregulated pilus colonization factor . In this study, we have characterized the 38-kDa OmpU protein and investigated its role in the adhesion of V . cholerae to mammalian cells . The amino-terminal sequence of OmpU has similarity with the sequences of Haemophilus influenzae HMW1 and HMW2 adhesins, which, in turn, also have similarity with the sequence of Bordetella pertussis filamentous hemagglutinin . A monoclonal antibody directed against FHA recognized both V . cholerae OmpU and Escherichia coli OmpA, and polyclonal anti-OmpU antibodies recognized FHA and E . coli OmpA, suggesting the existence of common epitopes among these proteins . OmpU was strongly recognized by convalescent-phase serum from volunteers experimentally infected with virulent V . cholerae strains, indicating that OmpU is immunogenic and produced in vivo . OmpU selectively bound to fibronectin and to an arginine-glycine-asparagine (RGD) tripeptide but not to other matrix glycoproteins tested such as collagen or laminin . Antibodies directed against OmpU or their F(ab)2 fragments completely inhibited adhesion of several V . cholerae strains to HeLa, HEp-2, Caco-2, and Henle 407 epithelial cells and also inhibited intestinal colonization and conferred protection in newborn mice against both biotypes (El Tor and classical) of V . cholerae O1 . Collectively, these data indicate that OmpU has adhesive properties which may play a role in the pathogenesis of cholera.

Infect Immun, 1995 Nov, 63(11), 4409 - 16
Characterization of the hemolytic activity of Haemophilus ducreyi; Totten PA et al.; H . ducreyi is the causative agent of chancroid, a genital ulcer disease most prevalent in developing countries . Chancroid enhances the heterosexual transmission of human immunodeficiency virus and is identified in focal outbreaks in the United States, but little is known about its pathogenesis . We studied the hemolysin produced by H . ducreyi because this molecule might be an important virulence factor in the pathogenesis of chancroid . Ten strains of H . ducreyi were tested on newly devised blood agar plates and were found to have hemolytic activity . We examined the hemolytic activity of H . ducreyi 35000 further and found that it was heat labile, cell associated, greatest at pH 7.0, and produced in logarithmic- but not stationary-phase cultures . Using transposons Tn916 and Tn1545-delta 3, we have isolated three classes of transposon mutants of strain 35000: those with no detectable hemolytic activity, those with reduced hemolytic activity, and those with enhanced hemolytic activity . Transposon insertions in the nonhemolytic mutants were located in a DNA sequence which hybridized to the Proteus mirabilis hemolysin gene . Analysis of clones containing overlapping sections of this region served to further localize the H . ducreyi hemolysin gene and allow its expression in Escherichia coli and complementation of the nonhemolytic defect in an H . ducreyi mutant . These experiments indicate that H . ducreyi 35000 produces a hemolysin that is related to the calcium-independent hemolysin produced by P . mirabilis . Further experiments are needed to define the similarity of the H . ducreyi hemolysin to other calcium-independent hemolysins and to determine its role in the pathogenesis of chancroid.

Infect Immun, 1995 Nov, 63(11), 4395 - 401
Mapping of a surface-exposed, conformational epitope of the P6 protein of Haemophilus influenzae; Bogdan JA Jr et al.; P6 is an outer membrane protein of Haemophilus influenzae that is antigenically conserved and considered a candidate component of future H . influenzae vaccines . P6 contains a surface-exposed epitope recognized by monoclonal antibody (MAb) 3B9 . This epitope has been shown to be distinct from that recognized by the P6-specific MAbs 7F3 and 4G4 in a competitive inhibition enzyme-linked immunosorbent assay (ELISA) . MAb 3B9 did not bind to synthetic P6-specific sequential and overlapping hexameric peptides . Five peptides made to correspond to P6 sequences with high probabilities of surface exposure did not inhibit binding of MAb 3B9 to P6 . An antiserum to one of the peptides, designated SP66, inhibited binding of MAb 3B9 to P6 . A rabbit antiserum to P6 bound to sequential hexameric peptides, Gly-87AsnThrAspGluArgGlyThr-94, which were in the SP66 region of P6 . This antiserum inhibited the binding of P6 to MAb 3B9 in a competitive inhibition ELISA . P6 mutations with His and Ala substitutions at residues Thr-88 and Asn-89 still bound MAb 3B9 . MAb 3B9 reacted with Escherichia coli OmpA and Salmonella typhimurium OmpA . Sequence comparisons of P6 with these proteins indicated that the residue in the SP66 region responsible for binding is either Gly-87, Asp-90, or Gly-93 . Mercaptoethanol reduction abolished MAb 3B9 binding to E . coli OmpA and S . typhimurium OmpA . In these proteins, immediately downstream of the second cysteine, there is an ArgArg dipeptide which is identical to and aligns with Arg-147Arg-148 in P6 . This dipeptide has a high probability of surface exposure in P6 . Mutagenesis of the Arg-147Arg-148 to an AlaAla dipeptide in P6 abolished binding of MAb 3B9, demonstrating that it was either a portion of the epitope or important in the protein folding necessary for expression of this epitope . This study demonstrates that MAb 3B9 recognizes a conserved conformational determinant on the surface of H . influenzae that is composed of two discontinuous regions of P6.

Infect Immun, 1995 Nov, 63(11), 4389 - 94
Distinct antigenic and genetic properties of the immunoglobulin A1 protease produced by Haemophilus influenzae biogroup aegyptius associated with Brazilian purpuric fever in Brazil; Lomholt H et al.; All examined Haemophilus influenzae biogroup aegyptius isolates of the clone associated with Brazilian purpuric fever (the BPF clone) produced type 2 immunoglobulin A1 (IgA1) proteases encoded by identical iga genes that were distinct from the iga genes of other Brazilian H . influenzae biogroup aegyptius isolates . A partial nucleotide sequence analysis revealed close similarities to the iga genes of H . influenzae serotype c and one noncapsular H . influenzae biotype III strain isolated from a case of conjunctivitis in Tunisia, suggesting an evolutionary relationship . Epitopes recognized by neutralizing antibodies differed for the IgA1 proteases of the BPF clone and of other H . influenzae strains, including Brazilian H . influenzae biogroup aegyptius isolates from patients with noninvasive conjunctivitis . The low probability of developing cross-reacting neutralizing antibodies to the IgA1 protease of the BPF clone may contribute to the pathogenic potential of this virulent phenotype in Brazil.

Infect Immun, 1995 Nov, 63(11), 4219 - 23
The V-region repertoire of Haemophilus influenzae type b polysaccharide antibodies induced by immunization of infants; Chung GH et al.; Haemophilus influenzae type b (Hib) is a significant pathogen for young children, and three Hib vaccines (named PRP-OMPC, HbOC, and PRP-T) are currently available for young children . Extensive studies of anti-Hib polysaccharide (PS) antibodies (Abs) have shown that the V regions of Abs against the Hib PS comprise a VH gene in the VH3 gene family and a VL gene from various K kappa and V lambda subgroups . To study immunogenic properties of the three vaccines in young children, we determined the VL subgroups and avidities of anti-Hib-PS Abs induced by the three clinically available conjugate vaccines . Ab avidity was measured by determining the concentration of a Hib-PS oligomer that abrogates half of the binding of immunoglobulin G anti-Hib-PS Abs to microwells . The PRP-OMPC vaccine induced lower-avidity Abs than the prelicensure HbOC vaccine (P = 0.05) . When we compared anti-Hib-PS Abs expressing V kappa Ia, V kappa II, and V lambda subgroups, a greater Ab response was induced by the prelicensure HbOC vaccine than other vaccines (P < 0.05) . When anti-Hib-PS Abs with the V kappa III subgroup were compared, however, both PRP-T and prelicensure HbOC vaccines induced a comparable response, which in turn was greater than those induced by the PRP-OMPC or the postlicensure HbOC vaccine (P < 0.001) . The VL repertoire of Abs induced with the prelicensure HbOC or PRP-T vaccine in young children is dominated (about 80%) by anti-Hib-PS Abs using subgroup V kappa II . However, anti-Hib-PS using V kappa II VL accounts for only about 40% of the total anti-Hib-PS Abs induced with the PRP-OMPC vaccine or the postlicensure HbOC . Our data suggest that immunogenic properties of Hib vaccines in young children vary depending on the vaccine preparations as well as the vaccine types.

Clin Exp Immunol, 1995 Nov, 102(2), 406 - 16
A possible role for lysozyme in determining acute exacerbation in chronic bronchitis; Taylor DC et al.; The aggregation of non-serotypable Haemophilus influenzae (NTHI) by whole saliva from patients with chronic obstructive lung disease (COLD) was investigated . Significant differences were observed between salivary aggregating activity of a control and COLD population (P < 0.001) . Saliva from patients less prone to acute exacerbations had a greater capacity to aggregate bacteria compared with saliva from patients with a predilection to infection . The mechanism of saliva-mediated aggregation of NTHI was investigated and shown to be related to lysozyme content . Lysozyme activity in saliva was measured by the turbidimetric technique and results showed that patients with chronic bronchitis had increased levels of salivary lysozyme, with a subpopulation within the non-infection-prone group having greater amounts . A significant difference was observed in salivary lysozyme between controls and non-infection-prone (P < 0.005) and infection-prone (P < 0.05) patients, respectively: the non-infection-prone patients having significantly (P < 0.005) more than the infection-prone patients . There was significant correlation (r = 0.742, P < 0.001) between salivary aggregation of NTHI and lysozyme activity . Chromatographically purified human lysozyme had a similar aggregation profile to that of saliva . There was no difference in serum and saliva lactoferrin concentrations between groups, but there was a significant increase (P < 0.05) in serum lysozyme concentration in the non-infection-prone group . This study suggests that the level of salivary lysozyme derived from macrophages may play an important role in determining resistance or susceptibility to acute bronchitis.

Clin Exp Immunol, 1995 Nov, 102(2), 243 - 50
In vitro stimulation of peripheral blood mononuclear cells (PBMC) from HIV- and HIV+ chancroid patients by Haemophilus ducreyi antigens; Van Laer L et al.; The cellular immune responses to fractionated Haemophilus ducreyi antigens, coated on latex beads, were assessed in patients with chancroid and in controls, using an in vitro lymphocyte proliferation assay . Several fractions of H . ducreyi antigen revealed stimulating activity . However, only the molecular size ranges 91-78 kD, 59-29 kD, and 25-21 kD induced proliferation that may be specifically related to H . ducreyi infection . Lymphocytes from four HIV- patients, successfully treated for chancroid, were not stimulated by H . ducreyi antigen . In general, lymphocytes from HIV+ chancroid patients were less responsive to H . ducreyi antigen compared with those from HIV- chancroid patients . However, two HIV-infected patients showed exceptionally strong responses to high molecular weight fractions . To our knowledge this is the first report demonstrating that H . ducreyi contains specific T cell-stimulating antigens . Based on this work, further identification and purification of the T cell antigens is feasible.

Am J Epidemiol, 1995 Nov 1, 142(9), 1000 - 6
Case-cohort analysis of case-coverage studies of vaccine effectiveness; Moulton LH et al.; Evaluation of vaccine field effectiveness may be performed by combining surveillance data on incident cases with an immunization coverage survey . Although many methods have been used for the analysis of studies of similar design, they are not always desirable or optimal . The authors discuss these approaches and propose use of a case-cohort analysis for such a study design when appropriate . The case-cohort analytic approach is illustrated with data from studies of a vaccine for Haemophilus influenzae type b (Hib) disease in children living on a southwestern Native American reservation during 1988-1993.

Arch Surg, 1995 Nov, 130(11), 1228 - 31; discussion 1231-2
Haemophilus pneumonia is a common cause of early pulmonary dysfunction following trauma; Spain DA et al.; BACKGROUND: Haemophilus species are a common cause of community-acquired pneumonia; however, their significance in posttraumatic pneumonia is unclear . DESIGN: Case series . SETTING: University hospital, level I trauma center . PATIENTS: Two hundred fifty-seven consecutive patients with blunt and penetrating trauma treated for pneumonia . MAIN OUTCOME MEASURES: Length of stay in the intensive care unit, duration of ventilatory support, rate of recurrent or persistent pneumonia, and mortality . RESULTS: Ninety-six (37%) of 257 patients treated for pneumonia had a Haemophilus species isolated on sputum culture . Of these 96 patients, 49 (51%) had only Haemophilus species, while 33 (34%) had associated gram-positive organisms and 14 (15%) had gram-negative organisms . Seventeen pure cultures (29%) and seven mixed cultures (15%) (P < .05) were beta-lactamase-positive trains . Compared with patients who had pneumonia caused by other bacteria, patients with Haemophilus species were younger (mean +/- SE, 35 +/- 1.7 vs 42 +/- 1.6 years; P < .05) and more severely injured (Injury Severity Score, 20.7 +/- 1.1 vs 17.5 +/- 0.9; P < .05) . There were no differences in any outcome variables between the two groups . Only one (1%) of 96 patients had persistent Haemophilus species on sputum cultures after 7 days of treatment . CONCLUSIONS: Haemophilus species are a frequent cause of pneumonia following traumatic injury . This occurs primarily in the early postinjury phase and therefore should be included in the differential diagnosis of early posttraumatic pulmonary insufficiency.

Am Fam Physician, 1995 Nov 1, 52(6), 1739 - 46
Resistant respiratory pathogens and extended-spectrum antibiotics; Pichichero ME; Traditional antibiotics such as amoxicillin, tetracycline and erythromycin remain the drugs of first choice for most bacterial respiratory infections . However, the usefulness of these agents varies, depending on local bacterial resistance patterns and patient factors . In the United States, amoxicillin and penicillin resistance currently occurs in 20 to 30 percent of Streptococcus pneumoniae strains, 30 to 40 percent of Haemophilus influenzae strains and 70 to 90 percent of Moraxella catarrhalis strains . For infections with these pathogens, selective use of the newer extended-spectrum oral antibiotics may be indicated . Cefuroxime axetil (a second-generation cephalosporin), cefpodoxime (a third-generation cephalosporin), amoxicillin-clavulanate (a beta-lactamase inhibitor combination agent) and clarithromycin or azithromycin (extended-spectrum macrolides) are all relatively effective against organisms that are commonly resistant to penicillin and amoxicillin.

Postgrad Med, 1995 Nov, 98(5), 141 - 4, 146, 149-50
Vaccination update . Hib, hepatitis, polio, varicella, influenza, pneumococcal and meningococcal disease; Vetter RT et al.; Immunization of infants and children is the most effective strategy for decreasing the incidence of some infectious diseases . Most invasive disease due to Haemophilus influenzae type b occurs before age 5 years, and routine vaccination of infants for hepatitis B is currently recommended because selective immunization of high-risk persons has not been feasible . Decades of use of poliovirus vaccine has effectively eliminated cases of wild-virus infection, although some vaccine-related cases still occur . The newly approved varicella vaccine appears to be a cost-effective way to decrease infection rates in children . Improved immunization rates for influenza and pneumococcal and meningococcal diseases could help decrease excess mortality in elderly persons and those with chronic illness.

Pediatrics, 1995 Nov, 96(5 Pt 1), 951 - 4
Effect of urine latex agglutination tests on the treatment of children at risk for invasive bacterial infection; Adcock PM et al.; BACKGROUND . Identifying febrile children with invasive bacterial infection is difficult in the absence of telltale physical findings . Urine latex agglutination (ULA) tests have been used for rapid, on-site identification of such children . OBJECTIVES . To study the performance of ULA tests in identifying children with Haemophilus influenzae, Streptococcus pneumoniae, and group B streptococcus infection and to examine how the results of ULA tests affect patient treatment . DESIGN . Retrospective review . SETTING . Urban children's hospital . PATIENTS . All emergency department and hospital patients tested by ULA in 1990, excluding patients in the neonatal units . RESULTS . Of 1629 patients, 36 had positive tests (20 H influenzae, 5 S pneumoniae, and 11 group B streptococcus) . Thirteen of these were false positive based on culture results . Although ULA tests demonstrated excellent specificity, their sensitivity was poor . Positive predictive values for bacteremia ranged from 0.346 to 0.600, and negative predictive values ranged from 0.972 to 0.997 . The decision to treat with antibiotics was made before ULA test results were available in 19 (53%) of the 36 patients with positive test results . Of 1593 patients with negative test results, 1211 (76%) were admitted to the hospital, and approximately 81% were empirically treated with parenteral antibiotics . CONCLUSIONS . In clinical practice, treatment and disposition decisions are frequently made without regard to ULA test results.

Leukemia, 1995 Nov, 9(11), 1902 - 9
Improved vaccination response during ranitidine treatment, and increased plasma histamine concentrations, in patients with B cell chronic lymphocytic leukemia; Jurlander J et al.; Patients with B cell chronic lymphocytic leukemia (B-CLL) have decreased capacity to mount relevant antibody responses upon immunization, and development of hypogammaglobulinemia is part of the natural history of the disease . We investigated the influence of histamine type-2 (H2) receptor blockade by ranitidine on the in vivo antibody production in B-CLL patients following vaccination . Anti-polysaccharide antibodies in B-CLL patients, vaccinated with a tetanus-toxoid conjugated vaccine against Haemophilus influenzae type-B (Hib), reached long-term protective levels in more than 90% of B-CLL patients randomized to ranitidine treatment, as compared to 43% of the untreated patients (P = 0.024) . No difference in the response to vaccination against influenza virus types A and B protein could be detected between the two groups . Plasma histamine levels were 2-fold to 20-fold higher in 23 out of 31 B-CLL patients, compared to normal controls, and these levels showed a significant positive correlation to disease duration . These findings indicate the possibility of improving in vivo antibody production against a highly relevant pathogen in B-CLL patients by histamine type-2 receptor blockade, and the combined finding of an immune-stimulatory effect of ranitidine and increased plasma histamine levels, strongly suggests the involvement of histamine in the pathogenesis of B-CLL immunodeficiency.

Science, 1995 Oct 20, 270(5235), 397 - 403
The minimal gene complement of Mycoplasma genitalium; Fraser CM et al.; The complete nucleotide sequence (580,070 base pairs) of the Mycoplasma genitalium genome, the smallest known genome of any free-living organism, has been determined by whole-genome random sequencing and assembly . A total of only 470 predicted coding regions were identified that include genes required for DNA replication, transcription and translation, DNA repair, cellular transport, and energy metabolism . Comparison of this genome to that of Haemophilus influenzae suggests that differences in genome content are reflected as profound differences in physiology and metabolic capacity between these two organisms.

J Biol Chem, 1995 Oct 20, 270(42), 25142 - 9
Biochemical characterization of a Haemophilus influenzae periplasmic iron transport operon; Adhikari P et al.; Bacterial iron transport is critical for growth of pathogens in the host environment, where iron is limited as a form of nonspecific immunity . For Gram-negative bacteria such as Haemophilus influenzae, iron first must be transported across the outer membrane and into the periplasmic space, then from the periplasm to the cytosol . H . influenzae express a periplasmic iron-binding protein encoded by the hitA gene . This gene is organized as the first of a three-gene operon purported to encode a classic high affinity iron acquisition system that includes hitA, a cytoplasmic permease (hitB), and a nucleotide binding protein (hitC) . In this study we describe the cloning, overexpression, and purification of the H . influenzae hitA gene product . The function of this protein is unambiguously assigned by demonstrating its ability to compete for iron bound to the chemical iron chelator 2,2'-dipyridyl, both in vitro and within the periplasmic space of a siderophore-deficient strain of Escherichia coli . Finally, the importance of a functional hitABC operon for iron acquisition is demonstrated by complementation of this siderophore-deficient E . coli to growth on dipyridyl-containing medium . These studies represent a detailed genetic, biochemical, and physiologic description of an active transport system that has evolved to efficiently transport iron and consequently is widely distributed among Gram-negative pathogenic bacteria.

Schweiz Rundsch Med Prax, 1995 Oct 3, 84(40), 1125 - 8
{Therapeutic problems in pneumonia}; Gonon M et al.; In Switzerland 10 out of 1000 adults suffer from pneumonia each year . It is of note that mycoplasma, influenza virus and pneumococci are the most common causative agents of community-acquired pneumonia . For the latter macrolides are presently the antibiotics of choice . Pneumonias occurring in patients with immune disorders should be treated primarily with amoxicillin + clavulanic acid or with cephalosporins of the second generation, because infestation with germs like haemophilus influenzae and klebsiella pneumoniae have to be considered . If the empirically chosen therapy should fail, the therapeutic strategy should not be changed blindly . Differential diagnosis and appropriate investigations are necessary (other germ, other disease, complications?) . Problems in treating patients with pneumonia are illustrated by three case examples.

Indian Pediatr, 1995 Oct, 32(10), 1077 - 82
Evaluation of immunogenicity and tolerance of single dose haemophilus influenzae type B (PRP-T) vaccine; Acharya D et al.; OBJECTIVE: Evaluation of immunogenicity and acceptability of PRP-T vaccine among the Indian children . DESIGN: Multicentric, open, parallel group, comparative study of Haemophilus influenzae type B vaccine, given as single (Group I) or associated (Group II) with DPT vaccine . SETTING: Five different vaccination clinics . SUBJECTS: 125 children between the age group of 18-24 months . PARAMETERS: Measurement of (i) pre and post vaccination antibody titres of Haemophilus influenze type B specific antibody; (ii) Adverse events; and (iii) Tolerance as graded by the physician . RESULTS: Prevaccination antibody levels were > 0.15 mcg/ml in 56.3% in Group I and 35.7% in Group II . Post-seroconversion was seen in 97% in Group II receiving single and all in Group II (P > 0.05) . The vaccine was well tolerated . CONCLUSIONS: The probability of subclinical infection or cross immunity is high in India . ACTHIB vaccine has a good immunogenicity and tolerance and association with DPT does not modify the immunogenicity of ACTHIB vaccine.

East Afr Med J, 1995 Oct, 72(10), 645 - 8
Low dose erythromycin regimen for the treatment of chancroid; Kimani J et al.; Haemophilus ducreyi is the commonest cause of genital ulcer disease in Africa and is associated with heterosexual transmission of human immunodeficiency virus(HIV) . The World Health Organization currently recommends erythromycin 500 mg three times a day for seven days as the treatment of choice for Haemophilus ducreyi infection . We studied the effectiveness of a lower dose erythromycin treatment regime, 250 mg three times a day for seven days in the treatment of chancroid . Patients with genital ulcer disease presenting at Nairobi City council clinic between January and March, 1992 were recruited into the study . Swabs were taken from the ulcers for Haemophilus ducreyi and venous blood was screened for syphilis and HIV antibodies . A total of 219 patients were enrolled for the study and were reviewed on days seven and fourteen for side effects, bacteriological and clinical cure rates . 26.4% of the study population were HIV-1 seropositive . The treatment regime was well tolerated and effective in both HIV seropositive and seronegative patients . Complete bacteriological cure rate was achieved in Haemophilus ducreyi culture positives by day seven irrespective of the HIV serostatus . However, the clinical cure rate for HIV seropositive patients was 88% compared to 99% for seronegative patients (p<.001) . It is concluded that a low dose erythromycin is an inexpensive and effective treatment for chancroid with complete bacteriological cure rate, although the healing process takes longer in HIV seropositive patients.

Aust J Public Health, 1995 Oct, 19(5), 465 - 70
A population-based survey of immunisation coverage in two-year-old children; Herceg A et al.; A cross-sectional, population-based, cluster-sample survey of 187 children was conducted in the Newcastle area to assess the proportion of two-year-old children who were fully immunised, to ascertain whether administration of these vaccines was age-appropriate and to look for factors predicting incomplete immunisation . Parents or guardians were interviewed at their homes and the immunisation status of the children was verified either by the parent-held record or by the immunisation-provider-held record . Levels of full immunisation were 77 per cent at the time of interview and 72 per cent at the second birthday if Haemophilus influenzae type b (Hib) vaccine was excluded . If it was included, the full immunisation level was 51 per cent . Coverage was highest for oral polio vaccine and lowest for Hib vaccine . Twenty-nine per cent of all immunisations were given early, 44 per cent were given on time and 20 per cent were given late . Doses of vaccines due at older ages were more likely to be given late or not at all . Factors predicting incomplete immunisation were: the principal caregiver being aged under 25 years, being born outside Australia, having post-secondary qualifications, being female and having more than one child in the household . Immunisation coverage levels were not high enough to protect against outbreaks of pertussis and measles and cases of Haemophilus influenzae type b . Immunisation providers should aim to increase coverage to protect the population against all vaccine preventable diseases, and aiming at high-risk groups could more effectively do this.

Braz J Med Biol Res, 1995 Oct, 28(10), 1065 - 8
Diagnosis of bacterial meningitis and septicemia by serum counterimmunoelectrophoresis; Alkmin MG et al.; We compare the results obtained by counterimmunoelectrophoresis in samples of serum and cerebrospinal fluid with microbiologic methods for 3,298 patients suspected of bacterial meningitis and/or septicemia at Instituto Adolfo Lutz, Sao Paulo, in a retrospective study of the period from July 1998 to July 1994 . Of the 415 patients (12.6% of the total cases studied) who were positive by the serum test, only 249 (7.6% of the total cases studied) were also positive when cerebrospinal fluid was assayed . Thus, 40% of the positives (5.6% of the total) were identifiable by analysis of serum but not of cerebrospinal fluid . Neisseria meningitidis accounted for 77.7% (129) and Haemophilus influenzae for 22.3% (37) of the positive results obtained only when serum was examined . These data show that although sensitivity and specificity of serum counterimmunoelectrophoresis are relatively low compared to cerebrospinal fluid counterimmunoelectrophoresis, the serum test is necessary to complement cerebrospinal fluid counterimmunoelectrophoresis data.

West Afr J Med, 1995 Oct-Dec, 14(4), 238 - 41
Open clinical trial of roxithromycin in patients of Plateau Hospitals, Jos in upper and lower respiratory tract infections; Okeke EN et al.; An open clinical study to assess the efficacy and tolerance of Roxithromycin 150 mg twice daily was carried out amongst Nigerian patients with upper and lower respiratory tract infections at Plateau Hospital Jos . Twenty-two patients aged between 13 and 86 years comprising of twelve women, seven men and three children completed the study . 18 (81.8%) had bronchopulmonary infections, 3 (13.6%) had tonsillitis and 1 (4.6%) had otitis media . Pathogens isolated included streptococcus Pneumonia (22.7%), Streptococcus pyogenes (13.6%), Bramhella Catarrhalis (9.1%), Haemophilus influenzae (9.1%), Staphylococcus Aureus (4.6%), Klebsiella species (4.6%), Pseudomonas Aeruginosa (4.6%) . There was 88.2% bacteriological cure and patients responded fast, with no major adverse reactions . Roxithromycin is therefore concluded to be an effective well tolerated drug for treatment of respiratory tract infections in Nigerians.

Eur J Clin Microbiol Infect Dis, 1995 Oct, 14(10), 892 - 8
Monoclonal antibodies against Haemophilus ducreyi lipooligosaccharide and their diagnostic usefulness; Ahmed HJ et al.; Monoclonal antibodies against the lipooligosaccharide of Haemophilus ducreyi were produced . Two of them, MAHD6 and MAHD7, were found to be relatively, although not absolutely, specific and reacted with nearly all strains of Haemophilus ducreyi tested: 59 of 60 and 60 of 60, respectively . The diagnostic usefulness of MAHD7 was assessed . Clinical specimens collected in Zambia from patients with genital ulcers were tested using indirect immunofluorescence (IF), enzyme immunoassay (EIA), the polymerase chain reaction (PCR) and bacterial culture . Compared with culture, IF had a sensitivity of 100%; compared with PCR, sensitivity was 89% . The corresponding figures for the EIA were 83% and 74%, respectively . The sensitivity of culture compared with PCR was 63% . The results suggest that IF on genital smears using MAHD7 might be an excellent tool for the diagnosis of chancroid in high-prevalence populations . However, further evaluation of the specificity of this test is needed.

J Antimicrob Chemother, 1995 Oct, 36 Suppl B, 59 - 72
Bacterial complications of transplantation: diagnosis and treatment; Donnelly JP; Bacterial complications develop mainly after transplantation during the period before engraftment takes place . Wound infections, urinary tract infection and pneumonia are the commonest complications of solid organ transplantation and generally involve Gram-negative bacilli and Staphylococcus aureus . However, Gram-positive cocci will predominate when selective oral antimicrobial prophylaxis is given as is frequently the case in bone marrow transplant recipients . Oromucositis, induced by total body irradiation or anthracyclines, result in more bacteraemia due to oral viridans streptococci . The use of central intravenous catheters leads to an increase in bacteraemia and infection due to coagulase-negative staphylococci . Patients requiring intensive care are also at risk of nosocomial infections including legionellosis . Once engraftment has occurred, there is much less risk of bacterial infection but patients remain vulnerable to the intracellular pathogens Listeria monocytogenes, non-typhoid salmonellae, Norcardia spp . and mycobacteria for as long as they require immunosuppression . Any rejection crisis must be treated aggressively with high-dose steroids or other agents which further undermine an already fragile immunity . In bone marrow transplant recipients, graft versus host disease and its treatment exerts a more profound effect on immunity and often coincides with cytomegalovirus infection which compromises the patient even further . Such patients are again at risk of infection with the same range of pathogens encountered during neutropenia since the oral mucosa, gut and catheter, if one is present, provide the same portals of entry . Immunosuppressive therapy, in some centres, is discontinued once the risk of graft versus host disease is reduced, although the reconstitution of the immune system is a lengthy process and there is a continued deficiency of IgG which renders patients unable to opsonise the encapsulated bacteria Streptococcus pneumonia and Haemophilus influenzae . In contrast to bone marrow transplant recipients, those with a solid organ transplant require life-long immunosuppression and so remain susceptible to infections with intracellular pathogens and, even with minimal immunosuppression, there will always be the risk that common bacteria will cause infection in unusual places and that uncommon organisms will be involved in apparently straightforward infections.

J Chemother, 1995 Oct, 7(5), 432 - 41
Comparative evaluation of the clinical and microbiological efficacy of co-amoxiclav vs cefixime or ciprofloxacin in bacterial exacerbation of chronic bronchitis; Cazzola M et al.; In an open randomized study 218 outpatients (159 males and 59 females) ranging between 18 and 85 years of age (mean 61.9) suffering from bacterial exacerbation of chronic bronchitis have been randomly treated: 79 with co-amoxiclav (amoxicillin 875 mg+clavulanic acid 125 mg) twice daily, 69 with cefixime (400 mg) once daily, and 70 with ciprofloxacin (500 mg) twice daily for an average period of 10 days . Before treatment start, 234 bacterial strains (105 Gram-positive and 129 Gram-negative) were isolated as the cause of exacerbation; the leading pathogens were Streptococcus pneumoniae and Haemophilus spp . Eradication rates at the end of treatment were 82.2% for the co-amoxiclav group, 77.6% for the cefixime group, and 81.2% for ciprofloxacin group . Clinical success (cure+improvement) was obtained in 90.8% of the cases treated with co-amoxiclav, in 80.9% for the cefixime group and in 85.7% of patients treated with ciprofloxacin . Seven adverse events (8.9%) of which 4 cases of diarrhea and 3 of itching, were recorded in the co-amoxiclav group . Eleven adverse events (14.7%) were recorded in the cefixime group including gastrointestinal disturbances in 6 patients and mild to moderate increase of liver function in 2 . Nine adverse events (12.9%) occurred in the ciprofloxacin group, including insomnia in 3 patients, gastrointestinal disturbances in 2, and serious increase of liver function tests in one patient . It can be concluded that there were no statistically significant differences among the three treatment groups . However, co-amoxiclav demonstrated a higher efficacy rate than cefixime and ciprofloxacin and was better tolerated . Therefore, it can be used as a first-choice drug in the treatment of exacerbation of chronic bronchitis.

J Vet Diagn Invest, 1995 Oct, 7(4), 476 - 80
Experimental reproduction of Haemophilus parasuis infection in swine: clinical, bacteriological, and morphologic findings; Vahle JL et al.; Haemophilus parasuis is a common cause of polyserositis and polyarthritis in swine . Little is known about the mucosal and systemic sites of replication and lesions which follow an aerosol exposure to H . parasuis . In this experiment 5-week-old cesarean-derived, colostrum-deprived (CDCD) pigs were inoculated intranasally with an inoculum containing 2 x 10(9) colony-forming units of H . parasuis . Two principals and one control pig were necropsied at 12, 36, 84, and 108 hours postinoculation (PI) and samples obtained for bacteriologic culture and microscopic examination . Inoculated pigs developed clinical signs of inappetence, reluctance to move, lameness, and a serous nasal discharge . Macroscopic findings included a fibrinous polyserositis and polyarthritis 36 hours PI which became progressively more severe at 84 and 108 hours PI . No lung lesions were grossly visible . Microscopic lesions included a mild purulent rhinitis at each post inoculation interval and fibrinous to fibrinopurulent synovitis and serositis at 36, 84, and 108 hours PI . A focal suppurative bronchopneumonia was observed in one pig examined at 36 hours PI . The nasal cavity and trachea were the only mucosal sites from which H . parasuis was reisolated . Haemophilus parasuis was isolated from the blood and systemic sites at 36, 84, and 108 hours PI . Findings presented indicated that intranasal inoculation of 5-week-old CDCD pigs with H . parasuis results in clinical signs and lesions of polyserositis and polyarthritis typical of field cases and is a useful model for the study of H . parasuis pathogenesis . The results also suggest that H . parasuis initially colonizes the nasal mucosa.

J Biochem (Tokyo), 1995 Oct, 118(4), 796 - 801
Purification and reaction mechanism of arylsulfate sulfotransferase from Haemophilus K-12, a mouse intestinal bacterium; Lee NS et al.; A novel type of sulfotransferase, arylsulfate sulfotransferase {EC 2.8.2.22}, was purified to homogeneity from Haemophilus K-12, a mouse intestinal bacterium . The purified enzyme (M(r) 290,000) is composed of four subunits (M(r) 70,000) . The best donor substrate was 4-methylumbelliferyl sulfate, followed by beta-naphthyl sulfate, p-nitrophenyl sulfate (PNS), and alpha-naphthyl sulfate . The best acceptor substrate was alpha-naphthol, followed by phenol and resorcinol . The apparent Km for PNS using phenol as an acceptor and that for phenol and resorcinol . The apparent Km for PNS using phenol as an acceptor and that for phenol using PNS as a donor substrate were determined to be 0.095 and 0.71 mM, respectively . One of the reaction products, p-nitrophenol inhibited the enzyme noncompetitively with respect to PNS, but competitively with respect to alpha-naphthol . The Ki values of PNP for PNS and alpha-naphthol were 0.89 and 0.12 mM, respectively . The other reaction product, alpha-naphthyl sulfate, inhibited the enzyme competitively with respect to PNS, but non-competitively with respect to alpha-naphthol . The Ki values of alpha-naphthyl sulfate for PNS and for alpha-naphthol were 2.72 and 1.7 mM . These results suggest that the sulfate transfer reaction proceeds according to a ping pong bi bi mechanism.

Minerva Pediatr, 1995 Oct, 47(10), 401 - 8
{Sequelae of bacterial meningitis in childhood: a study of hearing impairment}; Palla G et al.; Bacterial meningitis is an infection of the central nervous system involving quite a number of neurological sequelae the most common of which is hearing impairment . To assess the incidence of audiological deficit the authors evaluated retrospectively 20 patients, between 4 months and 11 years of age, observed at the Pediatric Clinic at the University of Pisa between 1988 and 1993 . A clinical-neurological examination and a complete auditory assessment (using BAEPs and impedence audiometry) has been performed in every patient between 1 and 18 months after they left hospital . 4 children had persisting neurological sequelae, 3 patients developed sensorineural hearing loss and one child visual impairment and seizures . Haemophilus Influenzae was responsible for 2 cases of sequelae and Streptococcus Pneumoniae for the other 2 cases . The number of days of illness before hospitalization and the institution of an antibacterial treatment, persistence of fever and deviation from the normal level of consciousness and persistence of neck rigidity were not correlated with the presence of sequelae . No correlation has been found between sensorineural hearing loss and the kind of antibacterial therapy . It's advisable that every child, following bacterial meningitis, should undergo a complete and repeated audiological assessment to detect any lesser impairments and/or unilateral losses that may damage the development of speech and language in any way.

Monaldi Arch Chest Dis, 1995 Oct, 50(5), 366 - 71
Acute bronchial infection in chronic obstructive pulmonary disease; Dorca J; Bacterial bronchial infection is a frequent cause of COPD exacerbation but not its only aetiology . Increased purulent expectorant appears to be its best indicator rather than fever, non-productive cough or dyspnoea . The clinician must try to recognize this condition rather than systematically prescribe empirical antibiotics . Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis are the major pathogens . Although atypical bacteria are not frequent, Chlamydia pneumoniae could play significant role . During the last years, new antibiotics, much more expensive than other regimens, are widely prescribed, often without a rational approach . In patients not already on antibiotics, sputum Gram stain is useful for deciding which patient should be treated and what would be the best anti-biotic . When it is not available, the chosen antibiotic must be at least active against three major pathogens according to the local susceptibility patterns . In patients not responding to the initial treatment, the consideration of its potential spectrum holes is then more useful than sputum examination.

J Trop Pediatr, 1995 Oct, 41(5), 308 - 10
Bacterial meningitis: still a cause of high mortality and severe neurological morbidity in childhood; Daoud AS et al.; Among 121 cases of bacterial meningitis (age 2 months to 12 years; mean, 35 months) treated over a 3-year period, Neisseria meningitidis was the most common pathogen (33 per cent), then Haemophilus influenzae (32 per cent) and Streptococcus pneumoniae (15 per cent) . In the H . influenzae group, 95 per cent were aged below 2 years . Overall mortality was 12 per cent: higher in the S . pneumoniae (17 per cent) and less common organism (21 per cent) groups . Neurological sequelae in 21 (20 per cent) of the 106 survivors included hearing impairment in 17 and quadriparesis in eight . Meningitis caused by S . pneumoniae contained a significantly higher proportion of children with neurological morbidity (P = 0.0128) . The addition of dexamethasone treatment during the third year produced an apparent but not significant trend towards less mortality (P = 0.7568), fewer neurological sequelae (P = 0.3401) and less hearing impairment (P = 0.3903) . Despite the availability of effective chemotherapy, bacterial meningitis will remain an important cause of high mortality and considerable morbidity.

Microbiology, 1995 Oct, 141 ( Pt 10), 2405 - 16
Sequence, genetic analysis, and expression of Actinobacillus pleuropneumoniae transferrin receptor genes; Gonzalez GC et al.; The tbpA and tbpB genes encoding the transferrin receptor proteins Tbp1 and Tbp2 from a serotype 7 strain of Actinobacillus pleuropneumoniae were cloned, sequenced, and expressed in Escherichia coli . The tbpB gene was preceded by putative promoter and regulatory sequences and was separated from the downstream tbpA gene by a 13 bp intercistronic sequence suggesting that the two genes may be coordinately transcribed . Determination of the nucleotide sequence of this region facilitated PCR amplification of the tbp region from a serotype 1 strain for comparative purposes . The deduced amino acid sequences of the Tbp1 proteins had regions of homology with Neisseria Lbp and Tbp1s and with TonB-dependent outer membrane (OM) receptors of E . coli . The deduced amino acid sequences of the Tbp2 proteins were nearly identical to those presented in previous studies . Upon high-level expression of the tbpA gene, a large proportion of the recombinant Tbp1 was found in inclusion bodies and could not be affinity-isolated with immobilized porcine transferrin . Most of the remaining expressed Tbp1 was present in the OM fraction, was expressed at the surface of E . coli cells, and retained binding activity that was specific for the C-lobe of porcine transferrin . Although recombinant Tbp2 was found in inclusion bodies during high-level expression, a significant proportion was associated with a novel OM fraction that appeared in sucrose density gradients which was distinct from the OM fraction containing recombinant Tbp1 . The recombinant Tbp2 was accessible at the surface yet was unable to bind porcine transferrin . In contrast to previous observations, the binding by recombinant Tbp2 was specific for the C-lobe of porcine transferrin . These results indicate that the A . pleuropneumoniae transferrin receptor proteins have similar properties to the receptor proteins in Neisseria spp . and Haemophilus influenzae, and that functional studies performed with recombinant receptor proteins need to consider differences in processing and export of these proteins when expressed in heterologous hosts.

J Clin Epidemiol, 1995 Oct, 48(10), 1245 - 50
Latex particle agglutination test in the diagnosis of Haemophilus influenzae type B, Streptococcus pneumoniae and Neisseria meningitidis A and C meningitis in infants and children; Camargos PA et al.; The knowledge of purulent meningitis etiology is essential in deciding the immediate therapy; in developing countries, however, the etiological agent identification does not reach 60% of the cases . A comparative study using the latex particle agglutination test (LPAT) in cerebrospinal fluid (CSF) for the diagnosis of meningitis due to Haemophilus influenzae type b, Streptococcus pneumoniae or Neisseria meningitidis A and C was carried out in Belo Horizonte MG, Brazil . CSF culture was used as a gold-standard . Two hundred and ninety-nine children, ranging from 3 months to 14 years of age, were included in the investigation . One hundred and forty-four presented a positive CSF culture for the above mentioned bacteria; the remaining presented meningitis due to other organisms (other bacteria or viral) or a normal CSF . The sensitivity and the specificity of LPAT was 95.7 and 100.0% for N . meningitidis C, 95.2 and 100.0% for H . influenzae type b and 86.5 and 100.0% for S . pneumoniae, respectively . When all three organisms were considered simultaneously, the sensitivity and the specificity was 93.0 and 100.0%, respectively . Taking into consideration a realistic estimate of disease prevalence in the community where the diagnostic test is being used, the positive predictive value and the posttest probability were estimated as 36.7 and 47.1% for children < 5 years and as 21.3 and 35.1% for children < 14 years of age, respectively . LPAT is a useful diagnostic test for meningitis due to the studied pathogens, especially in developing countries where laboratory facilities are limited.

J Bacteriol, 1995 Oct, 177(19), 5447 - 52
Isolation and characterization of a conserved porin protein from Helicobacter pylori; Doig P et al.; Helicobacter pylori is a causative agent of gastritis in humans and is correlated with gastric ulcer formation . Infections with this bacterium have proven difficult to treat with antimicrobial agents . To better understand how this bacterium transports compounds such as antimicrobial agents across its outer membrane, identification of porin proteins is important . We have recently identified a family of H . pylori porins (HopA to HopD) (M . M . Exner, P . Doig, T . J . Trust, and R . E . W . Hancock, Infect . Immun . 63:1567-1572, 1995) . Here, we report on an unrelated porin species (HopE) from this bacterium . This protein had a apparent molecular mass of 31 kDa and was seen to form 50- and 90-kDa aggregates that were designated putative dimeric and trimeric forms, respectively . The protein was purified to homogeneity and, with a model planar lipid membrane system, was shown to act as a nonselective pore with a single channel conductance in 1.0 M KCl of 1.5 nS, similarly to other bacterial nonspecific porins . An internal peptide sequence of HopE shared homology with the P2 porin of Haemophilus influenzae . HopE was also shown to be antigenic in vivo as assessed by sera taken from H . pylori-infected individuals and was immunologically conserved with both patient sera and specific monoclonal antibodies . From these data, it appears that HopE is a major nonselective porin of H . pylori . The implications of these findings are discussed.

Infect Immun, 1995 Oct, 63(10), 4115 - 20
Quantitation and biological properties of released and cell-bound lipooligosaccharides from nontypeable Haemophilus influenzae; Gu XX et al.; Nontypeable Haemophilus influenzae (NTHi) is a major pathogen causing otitis media in children . NTHi releases lipooligosaccharide (LOS) as outer membrane fragments during its growth . The release of LOS may play an important role in the pathogenicity of otitis media caused by this organism . The amounts of LOS in bacterial cells and growth media for five NTHi strains were determined by quantitative silver staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis . These strains were estimated to have 1.6 x 10(6) to 4.8 x 10(6) LOS molecules per bacterium . During a 3-day growth period, these NTHi strains released variable but significant amounts of LOS into the growth medium . Cells started to release detectable amounts of LOS into the medium at 2 to 5 h and continued to do so for up to 48 or 72 h . The concentrations of LOS in the culture supernatants released by these five strains were 10 to 55 micrograms/ml at 24 h and 40 to 100 micrograms/ml at 72 h, which was 34 to 189% of the cell-bound LOS concentration . The biological properties of released and cell-bound LOSs from two representative strains were compared . Released LOS showed an approximately 10-fold increase in inducing human monocytes to produce tumor necrosis factor alpha, interleukin 1 beta, and interleukin 6, a 13- to 28-fold increase in mouse lethal toxicity, and a 16- to 37-fold increase in the clotting of Limulus amebocyte lysate . These results suggested that released LOS or its inflammatory mediators play a more important role than the LOS in bacteria in the pathogenicity of otitis media caused by this organism.

Infect Immun, 1995 Oct, 63(10), 3846 - 50
Mapping of functional regions on the transferrin-binding protein (TfbA) of Actinobacillus pleuropneumoniae; Strutzberg K et al.; Actinobacillus pleuropneumoniae can use porcine transferrin as the sole source of iron . Two proteins with molecular masses of approximately 60 kDa (TfbA) and 110 kDa have been shown to specifically bind porcine transferrin; from the TfbA protein, three isoforms from A . pleuropneumoniae serotypes 1, 5, and 7 have been identified and characterized by nucleotide sequence analysis . Here we defined the transferrin-binding region(s) of the TfbA protein of A . pleuropneumoniae serotype 7 by TnphoA mutagenesis, random mutagenesis, and peptide spot synthesis . The amino-terminal half of the TfbA molecule, which has only 36% amino acid sequence identity among the three isoforms, was shown to be responsible for transferrin binding by TnphoA mutagenesis . This result was confirmed by analysis of six random mutants with decreased transferrin binding affinity . The subsequent analysis of overlapping 16-mer peptides comprising the amino-terminal half of the TfbA molecule revealed three domains of 13 or 14 amino acids in length with transferrin-binding activity . They overlapped, or were very close to, point mutations decreasing transferrin-binding ability . The first and third domains were unique to the TfbA protein of A . pleuropneumoniae serotype 7 . In contrast, the sequence of the second domain was present in almost identical forms (12 of 14 residues) in the TfbA proteins of A . pleuropneumoniae serotypes 1 and 5; in addition, a sequence consisting of functionally homologous amino acids was present in the otherwise completely distinct small transferrin-binding proteins of Neisseria gonorrhoeae (TbpB), N . meningitidis (Tbp2), and Haemophilus influenzae (Tbp2).

Infect Immun, 1995 Oct, 63(10), 3835 - 9
Potential role of nitric oxide in the pathophysiology of experimental bacterial meningitis in rats; Buster BL et al.; We have investigated the possible role of nitric oxide (NO) in the pathophysiology of bacterial meningitis (BM) by using the rat model of experimental BM . The nitrite concentration in cerebrospinal fluid (CSF) was used as a measure of NO production in vivo since NO rapidly degrades to nitrite and nitrate . Rats were inoculated intracisternally with live bacteria (5 x 10(6) CFU of Haemophilus influenzae type b strain DL42 or Rd-/b+/O2), with bacterial endotoxin (20 ng of DL42 lipooligosaccharide {LOS} or 200 ng of Escherichia coli lipopolysaccharide), or with a saline control vehicle . CSF samples were collected preinoculation and at the time of maximal alteration in blood-brain barrier permeability (BBBP) . CSF {nitrite} was quantified by measuring A550 after addition of the Greiss reagent and comparison to a standard curve of sodium nitrite . Rats inoculated with either DL42, Rd-/b+/O2, LOS, or lipopolysaccharide demonstrated a significantly elevated mean peak CSF {nitrite} (8.34, 15.62, 10.75, and 10.44 mM, respectively) versus the concentration prior to treatment and/or those in saline-treated animals (5.29 and 5.33 mM, respectively; P < 0.05 for each comparison) . We then determined if there was a correlation between CSF {nitrite} and percent BBBP (%BBBP) at various time points postinoculation with Rd-/b+/O2 . %BBBP was defined as the concentration of systemically administered 125I-labeled bovine serum albumin in the CSF divided by the level of 125I-labeled bovine serum albumin in serum multiplied by 100 . The mean %BBBP increased in tandem with the mean CSF {nitrite} (R = 0.84, P = 0.018), which peaked at 18 h in the absence of a change in the serum {nitrite} . Systemic administration of the NO synthase inhibitor N-nitro-L-arginine methyl ester demonstrated a significant reduction of mean CSF nitrite production (0.95 versus 6.0 mM in controls; P = 0.02) when administered intravenously to animals which had been inoculated intracisternally with 20 ng of LOS . Suppression of mean leukocyte pleocytosis (3,117 versus 11,590 leukocytes per mm3 in control LOS-challenged rats; P = 0.03) and mean alterations of BBBP (2.11 versus 6.49% in control LOS-challenged rats; P = 0.009) was observed concomitantly with decreased CSF {nitrite} . These results support the hypothesis that NO contributes to increased %BBBP in experimental BM.

Infect Immun, 1995 Oct, 63(10), 3809 - 15
Characterization of transferrin binding proteins 1 and 2 in invasive type b and nontypeable strains of Haemophilus influenzae; Gray-Owen SD et al.; Haemophilus influenzae has the ability to obtain iron from human transferrin via two bacterial cell surface transferrin binding proteins, Tbp1 and Tbp2 . Although a wide array of strains have been shown to express these receptor proteins, two studies have recently identified a series of isolates which appeared to lack the ability to bind transferrin . Included in this group were the members of a cryptic genospecies of nontypeable biotype IV strains which appear to possess a tropism for female urogenital tissues and are major etiologic agents of neonatal and postpartum bacteremia due to H . influenzae . The present study employed oligonucleotide primers specific for genes encoding the Tbp proteins of a type b biotype I strain of H . influenzae to probe the genomic DNAs of isolates from the previous studies . The tbpA and tbpB genes which encode Tbp1 and Tbp2, respectively, were detected in all of the strains tested either by PCR amplification directly or by Southern hybridization analysis . All of the strains displayed a transferrin binding phenotype, and affinity isolation of receptor proteins with transferrin-conjugated Sepharose recovered Tbp1 and/or Tbp2 from 11 of 14 strains, including 2 of the nontypeable biotype IV strains . In addition, all of the strains were capable of growing on human transferrin specifically, indicating that the mechanism of iron assimilation from transferrin is functional and is not siderophore mediated . These results confirm the presence of tbp genes in all of the invasive H . influenzae isolates characterized to date, suggesting that Tbp-mediated iron acquisition is important in disease which initiates from either the respiratory or urogenital mucosa.

Infect Immun, 1995 Oct, 63(10), 3751 - 8
Immunogenicity of synthetic peptides of Haemophilus influenzae type b outer membrane protein P1; Chong P et al.; To identify the B- and T-cell epitopes of P1 of Haemophilus influenzae type b, 13 peptides covering 90% of the protein were chemically synthesized . Mouse, guinea pig, and rabbit antisera raised against purified native P1 were tested for their reactivities against the peptides in peptide-specific enzyme-linked immunosorbent assays (ELISAs) . Six immunodominant linear B-cell epitopes were mapped to residues 103 to 137, 189 to 218, 248 to 283, 307 to 331, 384 to 412, and 400 to 437 of the mature P1 protein . When P1 peptides were screened for their reactivities with three human convalescent-phase serum specimens, peptides corresponding to residues 39 to 64, 226 to 253, and 400 to 437 reacted strongly with the antisera . Four regions (residues 39 to 64, 226 to 253, 339 to 370, and 400 to 437) contained murine T-cell epitopes . Rabbit antipeptide antisera were tested for their reactivities with the immunizing peptides and P1 protein by ELISA and immunoblots . All anti-P1 peptide antisera except those raised against peptide HIBP1-8 (residues 279 to 312) or HIBP1-8-keyhole limpet hemocyanin conjugate were shown to be specific for their respective immunizing peptides by ELISA . In addition, rabbit antisera raised against the synthetic peptides corresponding to residues 1 to 29, 39 to 64, 103 to 137, 189 to 218, 226 to 253, 248 to 283, 307 to 331, and 400 to 437 of the mature P1 protein recognized the P1 protein from both typeable and nontypeable isolates . These results suggest that these peptides contain epitopes highly conserved among typeable and nontypeable strains of H . influenzae . However, none of the antipeptide antisera have bactericidal activity, nor were they protective against H . influenzae type b in the infant rat model of bacteremia.

Chest, 1995 Oct, 108(4), 932 - 6
Clinical utility of blood cultures in adult patients with community-acquired pneumonia without defined underlying risks; Chalasani NP et al.; STUDY OBJECTIVE: We retrospectively examined the clinical utility of obtaining routine blood cultures before the administration of antibiotics in certain nonimmunosuppressed patients with community-acquired pneumonia (CAP) admitted to the hospital during 1991 . DESIGN: Retrospective review . SETTING: Grady Memorial Hospital (a county hospital primarily serving inner-city Atlanta) . PATIENTS OR PARTICIPANTS: Hospital discharge diagnosis listings identified 1,250 adults ( > or = 18 years old) with pneumonia . From this group of patients, we selected patients admitted to the hospital with (1) respiratory symptoms and a lobar infiltrate on chest radiograph that were present at the time of hospital admission, (2) two or more sets of blood cultures obtained within 48 h of hospital admission, and (3) absence of defined risk factors: HIV-related illness, malignancy, recent chemotherapy, steroid therapy, sickle cell disease, nursing home residence, or hospital stays within the past 14 days . MEASUREMENTS AND RESULTS: Five hundred seventeen patients (mean age, 52 years;: age range, 18 to 103 years) qualified . Of these 517 patients, 25 patients (4.8%) had growth in blood cultures considered contaminants while 34 (6.6%) had blood cultures positive for the following pathogens: 29 Streptococcus pneumoniae, 3 Haemophilus influenzae, and 1 Streptococcus pyogenes, 1 Escherichia coli . Antibiotic therapy was changed for 7 of the 34 patients with positive blood cultures (1.4% of study patients) . Antibiotic regimens were altered in 48 additional patients based on sputum culture, poor clinical response, and allergic reactions . CONCLUSIONS: Few blood cultures were positive for likely infecting organisms in adult patients with CAP without defined underlying risk factors . Furthermore, a total of $34,122 was spent on blood cultures at $66 per patient . In this carefully defined group of patients, blood cultures may have limited clinical utility and questionable cost-effectiveness.

Am J Respir Crit Care Med, 1995 Oct, 152(4 Pt 1), 1316 - 20
Bacterial infection in chronic obstructive pulmonary disease . A study of stable and exacerbated outpatients using the protected specimen brush; Monso E et al.; The lower airways of asymptomatic chronic obstructive pulmonary disease (COPD) patients can be colonized by bacteria, mainly Haemophilus influenza, Streptococcus pneumoniae, and Moraxella catarrhalis . However, the role of lower airway bacteria in stable and exacerbated COPD has not been well defined . To determine the importance of lower airway bacterial infection in COPD we studied 40 outpatients with stable COPD (Group A: age 61.1 +/- 9.9 yr; {mean +/- SD}; FEV1/FVC 51.7 +/- 12.5) and 29 outpatients with exacerbated COPD (Group B: age 63.4, SD 9.0 yr; FEV1/FVC 52.0, SD 9.6), using the protected specimen brush (PSB) for microbiology sampling . Group A consisted of outpatients with stable COPD having normal or near-normal chest X-rays, with clinical indications for performing fiber-bronchoscopy (pulmonary nodule, remote hemoptysis); Group B consisted of patients with exacerbated COPD who voluntarily accepted lower airway microbiology sampling . To avoid contamination by upper airway flora the PSB was used for bacterial sampling in all the cases and concentrations > or = 1,000 colony-forming units/milliliter (CFU/ml) were considered positive . Results were as follows: Group A: Lung function data in outpatients with stable COPD were lower than the reference values for this population (FVC 2.97 +/- 1.02 L, FVC% 71.4 +/- 22.4, FEV1 1.59 +/- 0.79 L, FEV1% 51.2 +/- 23.0) . Positive PSB cultures were obtained in 10 of 40 cases (25%), mainly of H . influenzae and S . pneumoniae . Two of 40 cases had positive cultures at concentrations > or = 10,000 CFU/ml (5.0%).(ABSTRACT TRUNCATED AT 250 WORDS)

Int J Exp Pathol, 1995 Oct, 76(5), 317 - 30
The cochlear lesion in experimental bacterial meningitis of the rabbit; Osborne MP et al.; Sensorineural hearing loss was studied in a rabbit model of experimental bacterial meningitis using electrophysiological and ultrastructural techniques . Hearing impairment was monitored by auditory brain-stem evoked responses (ABERs) and concomitant structural lesions were identified by both transmission (TEM) and scanning (SEM) electron microscopy . Meningitis was induced by intra-cerebrospinal fluid injection of either Escherichia coli (strain 2073 and type K-12) or Haemophilus influenzae type b . Auditory loss of approximately equal to 10 dB occurred in all rabbits by about 10 hours post infection and progressed in severity until by 20 h following infection, hearing losses up to and > 60 dB were obtained . At levels of hearing loss < 20 dB ultrastructural damage to the organ of Corti was barely detectable . With greater levels of hearing loss, patchy structural damage to hair cells, synaptic nerve terminals, supporting cells and inner spiral sulcus cells and cells of the stria vascularis was clearly evident . Bacteria were found in scala tympani, the basilar membrane, the organ of Corti, scala media, the spiral ligament and at the margin of the stria vascularis . Evidence of bleeding was found in some cochleas; erythrocytes were found in scala tympani, scala media, amongst hair cells and beneath the tectorial membrane . The results show that hearing loss is associated with bacterial invasion and damage to the organ of Corti and that the cause of hearing loss is likely to result from multiple lesions within the cochlea . Lesions to sensory cells almost certainly will produce permanent hearing loss . Lesions to supporting cells, nerve terminals and to stria vascularis may well produce only temporary hearing loss.

J Trauma, 1995 Oct, 39(4), 784 - 6
Lethal Waterhouse-Friderichsen syndrome in posttraumatic asplenia; Locker GJ et al.; We herein report a case of fulminant lethal Waterhouse-Friderichsen syndrome in an elderly female patient seven years after posttraumatic splenectomy . In contrast to various reports, this patient had not been vaccinated against Streptococcus pneumoniae, Neisseria meningitidis, or Haemophilus influenzae, respectively, although infections with these microorganisms are known to cause the main lethal diseases in asplenic patients . Again, we recommend obligatory vaccinations against the mentioned bacteria for it is known that this decreases the risk of fatal septic events in these patients . To optimize prevention, it is imperative to vaccinate patients undergoing splenectomy before discharge from hospital.

J Immunol Methods, 1995 Sep 25, 185(2), 225 - 35
Detection of Haemophilus ducreyi lipooligosaccharide by means of an immunolimulus assay; Hansen EJ et al.; A murine monoclonal antibody (MAb) directed against a surface-exposed epitope of the lipooligosaccharide (LOS) of Haemophilus ducreyi strain 35000 was shown to be reactive with all 37 strains of this pathogen tested in a colony blot-radioimmunoassay . The LOS epitope bound by this MAb appeared to be stably expressed by H . ducreyi growing in vitro . The use of this MAb in the immunolimulus system revealed that it could detect purified H . ducreyi LOS at a level of 25 pg/ml . Similarly, this immunolimulus system could detect as few as 1000 colony forming units of in vitro-grown H . ducreyi cells per ml of buffer . When this MAb was utilized in the immunolimulus system together with lesion material from rabbits infected with two different H . ducreyi strains, a positive reaction was obtained with every sample tested, even when no viable organisms were present in the lesion material . In contrast, this MAb yielded consistently negative results when used in the immunolimulus system with lesion material from animals infected with Staphylococcus aureus.

Med J Aust, 1995 Sep 4, 163(5), 239 - 42
Antibiotic resistance among respiratory pathogens in preschool children; Hammond ML et al.; OBJECTIVE: To determine the prevalence of antibiotic resistance among common respiratory pathogens circulating in the community . DESIGN: Survey of common respiratory pathogens isolated from nasal discharges . SETTING: 117 childcare centres and kindergartens in metropolitan Melbourne between May and July 1991-1993 and 42 from sociodemographically matching suburbs in Sydney between May and July, 1993 . SUBJECTS: Children aged six years and under with nasal discharge . OUTCOME MEASURES: Resistance to penicillins, erythromycin and tetracycline among isolates of Haemophilus influenzae, Streptococcus pneumoniae, Moraxella (Branhamella) catarrhalis and Staphylococcus aureus . RESULTS: A total of 2286 nasal discharge swabs were collected . Amoxycillin resistance was detected in 99 of 711 isolates of H . influenzae (13.9%) and penicillin resistance in 781 of 834 isolates of M . catarrhalis (93.6%), 342 of 375 isolates of S . aureus (91.2%), and 30 of 781 isolates of S . pneumoniae (3.9%) . Of 86 strains of H . influenzae type b isolated, 20 (23.3%) produced beta-lactamase . Penicillin resistance tended to become more common among isolates of H . influenzae and S . pneumoniae during the three-year period . CONCLUSION: Antibiotic resistance, mediated by beta-lactamase or altered penicillin-binding proteins, among respiratory pathogens carried by preschool children was significant and possibly increasing . This highlights the impact of prescribed antibiotics in the community and the folly of prescribing the limited store of antibiotics for viral infections.

J Antimicrob Chemother, 1995 Sep, 36(3), 545 - 9
The effects of low concentrations of antibiotics on epithelial damage caused by non-typable Haemophilus influenzae and bacterial morphology; Tsang KW et al.; Sterile culture filtrates from non-typable Haemophilus influenzae (NTHi) grown in medium containing no antibiotics or 0.25 MIC of amoxycillin, ciprofloxacin or loracarbef were examined for their effect on the ciliary beat frequency (CBF) and structure of human respiratory epithelium . CBF slowing was significantly (P < 0.05) less with 0.25 MIC of all three antibiotics . The epithelium was significantly (P < 0.05) less disrupted with ciprofloxacin . The morphology of NTHi infecting human adenoid organ cultures after 24 h with or without 0.25 MIC of the same antibiotics was measured by scanning electron microscopy . Only ciprofloxacin caused a significant (P < 0.05) change in morphology.

Ann Ig, 1995 Sep-Oct, 7(5), 329 - 38
{Use of the economic evaluation in the comparison of different vaccination programs against Haemophilus influenzae type b disease}; de Campora E et al.; Although economic evaluation offers a relevant contribution to the health policy's choices, only few studies appeared so far on its application to the field of vaccinations . The present study compares several hypotheses of vaccination strategies in order to evaluate the opportunity to introduce a vaccination program against Haemophilus influenzae b infection . The vaccination strategies are evaluated in terms of costbenefits, cost-effectiveness and cost-utility . The results show that the highest benefits could be obtained performing mass vaccination in the age-group 0-5 years . Actually this hypothesis, compared to other, offers the best compliance and externalities; furthermore it has the highest ethical value.

Ann Ig, 1995 Sep-Oct, 7(5), 319 - 27
{Incidence of Haemophilus influenzae type b meningitis in Italy . Preliminary results}; D'Alessandro D et al.; The Authors describe the survey's results aimed at quantifying the Haemophilus influenzae type b meningitis incidence rate in Italy . The survey collected all the cases of 1994 notified per region to the Ministry of Health; furthermore, regional incidence rates are compared and integrated with other data obteined from local ad hoc studies . The results obtained show a high incidence in the age group 0-2 years and a large discrepancy in the incidence rate between regions (from 0.0 to 16.8 cases per 100,000); furthermore, in the same age group, for a large amount of meningitis cases (18% of the total) the aetiological diagnosis was not successful; this phaenomenon is certainly one of the causes for the underestimation of the disease . The integration with data collected in longitudinal studies, increases largely the regional incidence rate, showing the highest rate in the Lazio Region (18.5 cases per 100,000) . The Authors conclude by stressing the need to increase the cooperation between the Regional Health Authorities and the Ministry of Health, in order to obtain more reliable figures of the national incidence rates.

Clin Ther, 1995 Sep-Oct, 17(5), 875 - 81
Results of a survey of children with acute bacterial conjunctivitis treated with trimethoprim-polymyxin B ophthalmic solution; Wagner RS; Acute conjunctivitis, one of the most frequently seen eye diseases in infants and children, is associated with a shorter duration of clinical disease when antimicrobial agents are used . Although viruses often are implicated as causative agents, Haemophilus influenzae and Streptococcus pneumoniae are the most commonly isolated bacterial pathogens . Empiric therapy of acute conjunctivitis therefore should include agents with both gram-positive and gram-negative antimicrobial activity . Trimethoprim-polymyxin B is a broad-spectrum antimicrobial agent available as an ophthalmic solution . We conducted a patient outcomes study to evaluate the subjective response to treatment with trimethoprim-polymyxin B of children with presumed acute bacterial conjunctivitis . Questionnaires were distributed to more than 100 pediatricians who assessed outcome measures in 472 children with acute bacterial conjunctivitis for whom they prescribed trimethoprim-polymyxin B . The parameters evaluated were clinical outcome, overall efficacy, and comfort provided by the medication regimen . The physicians reported that 95% of the infected eyes were cured or improved within 7 days . In addition, the overall efficacy trimethoprim-polymyxin B was rated as excellent or good in 76% and 20% of cases, respectively . With regard to patient comfort, patients or their caregivers reported that patients were very comfortable or moderately comfortable in 62% and 27% of cases, respectively . Four adverse events were reported; all were transient and of mild-to-moderate intensity . The survey results support clinical research findings on the comparative efficacy and safety of trimethoprim-polymyxin B ophthalmic solution compared with other ophthalmic antimicrobial agents . The pediatricians in our survey who prescribed trimethoprim-polymyxin B ophthalmic solution for children with presumed acute bacterial conjunctivitis reported that this medication was effective and well tolerated.

Clin Ther, 1995 Sep-Oct, 17(5), 861 - 74
Clinical comparison of cefuroxime axetil and amoxicillin/clavulanate in the treatment of patients with secondary bacterial infections of acute bronchitis; Henry D et al.; Two independent, investigator-blinded, multicenter, randomized clinical trials compared the clinical and bacteriologic efficacy and safety of two oral antibiotics, cefuroxime axetil and amoxicillin/clavulanate, in the treatment of patients with secondary bacterial infections of acute bronchitis (hereafter denoted acute bronchitis) . Three hundred sixty patients with signs and symptoms of acute bronchitis were enrolled at 22 centers and were randomly assigned to receive 10 days of treatment with either cefuroxime axetil 250 mg twice daily (BID) (n= 177) or amoxicillin/clavulanate 500 mg three times daily (TID) (n = 183) . Patients were assessed for both clinical and bacteriologic responses once during treatment (at 3 to 5 days) and twice after treatment (at 1 to 3 days and at 13 to 15 days) . Bacteriologic assessments were based on sputum specimen cultures obtained before treatment and, when possible, after treatment . Organisms were isolated from the pretreatment sputum specimens of 162 (45%) of 360 patients, with the primary pathogens being Haemophilus influenzae, Haemophilus parainfluenzae, Streptococcus pneumoniae, Moraxella catarrhalis, and Staphylococcus aureus (28%, 25%, 11%, 9%, and 8% of isolates, respectively) . Thirty-four percent of the H influenzae isolates and 94% of the M catarrhalis isolates that were tested for beta-lactamase production were positive . A satisfactory clinical outcome (cure or improvement) was achieved in 86% (117 of 136) and 83% (130 of 157) of the clinically assessable patients treated with cefuroxime axetil or amoxicillin/clavulanate, respectively (P = 0.45) . With respect to the eradication of bacterial pathogens, a satisfactory outcome (cure, presumed care, or cure with colonization) was obtained in 91% (53 of 58) and 86% (60 of 70) of bacteriologically assessable patients treated with cefuroxime axetil or amoxicillin/clavulanate, respectively (P = 0.32) . Treatment with amoxicillin/clavulanate was associated with a significantly higher incidence of drug-related adverse events than was treatment with cefuroxime axetil (39% vs 23%; P = 0.001), primarily reflecting a higher incidence of drug-related gastrointestinal adverse events (37% vs 15%; P < 0.001), particularly diarrhea and nausea . Four patients in the cefuroxime axetil group and eight patients in the amoxicillin/clavulanate group withdrew from the study because of drug-related adverse events . These results indicate that cefuroxime axetil 250 mg BID is as effective as amoxicillin/clavulanate 500 mg TID in the treatment of patients with acute bronchitis but produces fewer gastrointestinal adverse events, particularly diarrhea and nausea.

Clin Ther, 1995 Sep-Oct, 17(5), 838 - 51
Clinical comparison of cefuroxime axetil suspension and amoxicillin/clavulanate suspension in the treatment of pediatric patients with acute otitis media with effusion; Gooch WM 3rd et al.; Two independent, investigator-blinded, multicenter, randomized clinical trials, one of which included microbiologic evaluation of middle-ear fluid obtained by use of tympanocentesis, compared the efficacy and safety of two oral antibiotics, cefuroxime axetil suspension and amoxicillin/clavulanate suspension, in the treatment of children 3 months to 12 years old diagnosed with acute otitis media with effusion (AOME) . Four hundred seventy-seven pediatric patients with signs and symptoms of AOME were enrolled at 20 centers and were randomly assigned to receive 10 days of treatment with either cefuroxime axetil suspension 30 mg/kg per day in two divided doses (n = 235) or amoxicillin/clavulanate suspension 40 mg/kg per day in three divided doses (n = 242) . Patients were assessed for their response to treatment once during treatment (at 3 to 5 days) and twice after treatment (at 1 to 4 days and at 14 to 18 days) . In the study that included tympanocentesis, bacteriologic assessments were based on middle-ear fluid cultures obtained pretreatment, and, when possible, posttreatment in patients with an unsatisfactory clinical outcome . Organisms were isolated from the pretreatment middle-ear fluid specimens of 120 (73%) of 164 patients undergoing tympanocentesis, with the primary pathogens being Streptococcus pneumoniae . Haemophilus influenzae, and Moraxella catarrhalis (27%, 24%, and 6% of isolates, respectively) . Forty-four percent of the H influenzae isolates and 94% of the M catarrhalis isolates that were tested for beta-lactamase production were positive . A satisfactory clinical outcome (cure or improvement) was obtained in 70% of clinically assessable patients treated with cefuroxime axetil or amoxicillin/clavulanate, respectively (P = 0.40) . With respect to the eradication of bacterial pathogens, in the study that included tympanocentesis a satisfactory outcome (cure or presumed cure) was obtained in 84% (32 of 38) and 95% (36 of 38) of bacteriologically assessable patients treated with cefuroxime axetil or amoxicillin/clavulanate, respectively (P = 0.26) . Treatment with amoxicillin/clavulanate was associated with a significantly higher incidence of drug-related adverse events than was treatment with cefuroxime axetil (37% vs 16%; P < 0.001), primarily reflecting a higher incidence of drug-related gastrointestinal adverse events (34% vs 12%; P < 0.001), particularly diarrhea . Eight patients in the cefuroxime axetil group and 11 patients in the amoxicillin/clavulanate group withdrew from the studies because of drug-related adverse events . These results indicate that cefuroxime axetil suspension 15 mg/kg twice daily is as effective as amoxicillin/clavulanate suspension 13.3 mg/kg three times daily in the treatment of pediatric patients with AOME, but produces fewer gastrointestinal adverse events, particularly diarrhea.

Clin Ther, 1995 Sep-Oct, 17(5), 800 - 10; discussion 779
A review of the causes and treatment of bacterial and allergic conjunctivitis; Friedlaender MH; The most common causes of ocular inflammation are allergic or infectious in origin . A presumptive diagnosis can often be made through a comprehensive patient history and evaluation of presenting signs and symptoms, although the constellation and intensity of clinical findings may vary . Patients with allergic conjunctivitis often have itchy, red eyes, whereas patients with bacterial conjunctivitis often give a history of morning crusting and difficulty opening the eyelids . The treatment of patients with allergic conjunctivitis includes avoiding the offending allergen and applying topical antihistamines, mast cell stabilizers, or nonsteroidal anti-inflammatory agents . Streptococcus pneumoniae and Haemophilus influenzae are responsible for most cases of bacterial conjunctivitis in children . Staphylococcus species is the predominant organism in adults . Therefore, the treatment of patients with bacterial conjunctivitis consists of an antimicrobial agent with a broad spectrum of activity against most susceptible pathogens . Other causes of inflammation need to be considered in patients with atypical clinical signs and symptoms and in patients who do not respond to presumptive therapy.

Electrophoresis, 1995 Sep, 16(9), 1657 - 8
A region of Haemophilus influenzae Rd hybridizes with repetitive human DNA; Mody RB et al.; A 17.7 kbp EcoR1 fragment of Haemophilus influenzae Rd (wild-type strain) DNA containing repeats has been cloned together with the DNA repair gene uvr3 . The region containing repetitive DNA was subcloned on a 13 kbp EcoR1 fragment (pHi13) . The insert hybridizes with restriction enzyme digests of H . influenzae Rd and H . parainfluenzae and exhibits a repetitive pattern . Discrete bands superimposed upon a polydisperse background were observed when pHi13 hybridizes with restriction digests of human DNA . The hybridization patterns indicate that H . parainfluenzae and human DNA contain cross-hybridizing sequences . The repeats of pHi13 are present on a 2.6 kbp PvuII fragment.

Ir Med J, 1995 Sep-Oct, 88(5), 160 - 1
Notification of meningococcal disease and Haemophilus influenzae meningitis: a requirement for public health action; Fogarty J et al.; ------------------------------------------------------------------------ ------------------------------------------------------------------------ ------------------------------------------------------------------------ ---------------------------meningitis was ascertained for the years 1987-1991 . Laboratory record review identified positive blood culture and cerebro-spinal fluid isolates for 4 Dublin hospitals serving the paediatric population of the Eastern Health Board . Infectious disease registers, inspected for evidence of notification, revealed than only 44% of culture positive cases had been notified . The notification rate for Haemophilus meningitis (33%) was significantly lower than that for meningococcal disease (53%) . All laboratory identified cases should have been notified . Public health assessment is required for each condition as rifampicin prophylaxis can prevent secondary cases in close contacts of both diseases . Improved notification is essential, not just to comply with the law but more importantly, for better management of these diseases.

Eur Respir J, 1995 Sep, 8(9), 1543 - 7
Risk factors for the development of Haemophilus influenzae pneumonia in hospitalized adults; Barreiro B et al.; Potential risk factors for developing Haemophilus influenzae nosocomial pneumonia have not been sufficiently studied . We wanted to investigate the incidence and risk factors for the development of Haemophilus influenzae pneumonia in the hospital by means of a multivariate analysis . A total of 468 cases of nosocomial pneumonia were observed during the study period, 317 (68%) of which were aetiologically diagnosed by means of highly reliable methods, and H . influenzae was isolated in 57 of them . Fifty of the 57 episodes of H . influenzae pneumonia occurred in mechanically-ventilated patients . Underlying diseases were: medical in 12 cases, surgical in 15 cases, and traumatological in 22 . Variables associated with Haemophilus influenzae nosocomial pneumonia in intubated patients after the univariate analysis were: "period between admission and pneumonia 2-7 days" and "no previous antibiotics" . A multivariate analysis demonstrated that the variables "no previous antibiotics" and "coma on admission" were risk factors for H . influenzae pneumonia . In nonintubated patients, no risk factors were found for H . influenzae pneumonia compared with other nosocomial pneumonia . We conclude that H . influenzae was involved in 57 out of 317 (18%) of nosocomial pneumonia registered in our institution, and the majority of patients (50 out of 57) were mechanically-ventilated . In this particular subgroup, coma of the patient on admission to hospital and absence of antibiotic treatment prior to developing pneumonia constitute two definite risk factors for developing H . influenzae nosocomial pneumonia.

Eur Respir J, 1995 Sep, 8(9), 1451 - 7
Effect of erythromycin on Haemophilus influenzae endotoxin-induced release of IL-6, IL-8 and sICAM-1 by cultured human bronchial epithelial cells; Khair OA et al.; Although several studies have demonstrated that low-dose, long-term erythromycin treatment is effective in the management of patients with chronic lower respiratory tract infections, such as chronic bronchitis, bronchiolitis and bronchiectasis, the mechanisms underlying the action of erythromycin are not clear . We have cultured human bronchial epithelial cells (HBEC) as explant cultures from surgical tissue, and have investigated the effect of erythromycin on H . influenzae endotoxin (HIE)-induced release of inflammatory mediators in these cultures . Confluent epithelial cell cultures were incubated with 100 micrograms.mL-1 HIE +/- 0.1-10 micrograms.mL-1 erythromycin and were investigated for interleukin-6 (IL-6), interleukin-8 (IL-8) and soluble intercellular adhesion molecule-1 (sICAM-1) released into the culture medium after 24 h . HIE significantly increased the release of IL-6 from 3.9 +/- 1.5 pg.micrograms-1 cellular protein (in control untreated cultures) to 12.1 +/- 1.5 pg.micrograms-1 cellular protein, and IL-8 from 83.7 +/- 8.2 pg.micrograms-1 cellular protein (in control cultures) to 225.7 +/- 44.8 pg.micrograms-1 cellular protein . Similarly, HIE led to a significantly greater release of sICAM-1 from 0.04 +/- 0.01 ng.microgram-1 cellular protein, in control cultures, to 3.8 +/- 0.9 ng.microgram-1 cellular protein . Incubation of the epithelial cultures in the presence of 0.1-10 micrograms.mL-1 erythromycin significantly blocked the HIE-induced release of IL-6, IL-8, and sICAM-1, at all concentrations of erythromycin investigated . Erythromycin also attenuated neutrophil chemotaxis and adhesion to human endothelial cells, mediated by incubation with conditioned medium obtained from HIE-exposed epithelial cell cultures, in vitro . These results suggest that H . influenzae-induced release of inflammatory mediators from airway epithelial cells could contribute to chronic airway inflammation, and that this effect may be modulated by treatment with erythromycin.

Pediatr Infect Dis J, 1995 Sep, 14(9), 782 - 6
Cerebrospinal fluid shunt infections in children; Ronan A et al.; We reviewed cerebrospinal fluid (CSF) shunt infections treated in the Royal Children's Hospital, Melbourne from 1981 to 1991 . Forty-one episodes of CSF shunt infection were found after 900 shunt operations, an infection rate of 4.5% . Clinical symptoms were nonspecific in 31.7% of episodes, and in 17.1% of episodes the initial CSF sample was normal on microscopy and biochemistry, although a pathogen was isolated on culture . Most episodes occurred within 4 months of the last operation on the shunt, the exception being infections caused by Haemophilus influenzae . Four patients died during treatment, but none could be attributed to infection alone . Treatment of suspected CSF shunt infection should not be withheld because of lack of firm clinical diagnosis or normal CSF microscopy, and episodes occurring more than 4 months after the last operation on the CSF shunt should receive antibiotic cover for H . influenzae.

Clin Diagn Lab Immunol, 1995 Sep, 2(5), 563 - 8
Evidence obtained with monoclonal antibodies that O antigen is the major antigen responsible for the cross-reactivities between serotypes 4 and 7 of Actinobacillus (Haemophilus) pleuropneumoniae; Rodriguez Barbosa JI et al.; Monoclonal antibodies (MAbs) against Actinobacillus (Haemophilus) pleuropneumoniae serotype 4 (reference strain M62 and field isolate F6) were produced and characterized . Three hybridoma clones were raised against strain M62, and 13 were raised against strain F6 . The predominant antibody class was immunoglobulin M (IgM), although IgG2a and IgG2b were also obtained . Three of the MAbs produced to field isolate F6 (5C5, 1E10, and 5H7) did not recognize the reference strain of serotype 4, another (6F7) was reactive with both reference strains of serotypes 4 and 7, and the remaining 12 MAbs reacted only with the reference strain of the homologous serotype . All epitopes recognized by MAbs, except for one (6F7), were sensitive to periodic acid oxidation, and all of them were resistant to boiling in the presence of sodium dodecyl sulfate and reducing conditions, as evidenced by immunodot . Enhanced chemioluminiscence-immunoblot assays revealed that 10 MAbs (3E12, 5B8, 7C3, 6F7, 7F5, 7E6, 5G4, 4F1, 7E10, and 4B8) recognized a ladder-like banding pattern, which is in accordance with the O side chain antigen of lipopolysaccharide (LPS), while the remaining 6 MAbs (5C5, 5H7, 1E10, 6D11, 6B4, and 5E4) blotted with high-molecular-weight regions composed of a single banding pattern . The suitability of MAbs for serotyping of 78 field isolates was also examined . A high correlation was found between the results previously established by indirect hemagglutination with polyclonal sera and those obtained by enzyme-linked immunosorbent assay with MAbs . According to the different immunoreactivity of MAbs, three groups were established: group I (MAbs 3E12, 5B8, 7C3, 6F7, and 7F5), group II (MAbs 7E6, 5G4, 4F1, 7E10, and 4B8), and group III (MAbs 5C5, 5H7, and 1E10) . MAbs 6D11, 6B4, and 5E4 could not be included in any of the described above . At least six different immunodominant epitopes on the O antigen of the A . pleuropneumoniae serotype 4 LPS were identified . Finally, the implications of the effect of the O antigen of LPS in cross-reactions between serotypes 4 and 7 are clearly evidenced.

Infect Agents Dis, 1995 Sep, 4(3), 153 - 60
Immunoglobulin gene usage in the human anti-pathogen response; Newkirk MM et al.; The human antibody response to foreign pathogens is generated to a relatively small number of target surface proteins and carbohydrates that nonetheless have an extensive array of epitopes . The study of human monoclonal antibodies to different pathogens shows that there are a diversity of mechanisms used to generate a sufficient repertoire of antibodies to combat the invading pathogens . Although many different immunoglobulin gene elements are used to construct the anti-pathogen response, some elements are used more often than would be expected if all elements were used randomly . For example, the immune response to Haemophilus influenzae polysaccharide appears to be quite narrow, being restricted primarily to a specific heavy-chain gene, 3-15, and a lambda light-chain family II member, 4A . In contrast, for the immune response to cytomegalovirus proteins, a wider group of gene elements is needed . It is also surprising that despite an investigator bias for IgG- rather than IgM-secreting immortal B cells (because of their high affinity and neutralizing abilities), 26% of light chains and 13% of heavy chains showed a very low level of somatic mutation, equivalent to an IgM molecule that has not undergone affinity maturation . Although some highly mutated IgG molecules are present in the anti-pathogen response, most of the monoclonal antibodies specific for viruses or bacteria have a level of somatic hypermutation similar to that of the adult IgM repertoire . A number of studies have shown that there are similarities in the antibody responses to pathogens and to self (autoantibodies).(ABSTRACT TRUNCATED AT 250 WORDS)

Antimicrob Agents Chemother, 1995 Sep, 39(9), 2150 - 2
Effect of cefotaxime or ceftriaxone treatment on nasopharyngeal Haemophilus influenzae type b colonization in children; Goldwater PN; The effects of cefotaxime and ceftriaxone treatment on nasopharyngeal carriage of Haemophilus influenzae type b (Hib) were prospectively studied with 53 children with invasive Hib disease . Nasopharyngeal aspirates were monitored during therapy . Hib was eliminated within 2 days in 92% of patients and was eliminated in all patients after the third day of antibiotic treatment.

Ann Trop Paediatr, 1995 Sep, 15(3), 243 - 7
Pyogenic meningitis in children in north-western Ethiopia; Gedlu E et al.; A 5-year retrospective study of all children with acute pyogenic meningitis admitted to a district hospital in north-western Ethiopia was carried out from 1990 to 1994 . A total of 132 cases of pyogenic meningitis were identified . The causative bacteria were identified in 85 (64%) patients . The most common pathogen was Haemophilus influenzae (40%) with a case fatality rate of 29.4%, followed by Neisseria meningitidis (36.4%), which had a case fatality rate of 16.1%, and Streptococcus pneumoniae (20%) with a case fatality rate of 35.3% . Boys accounted for 64% of the patients, giving a male:female ratio of 1.7:1 . Median patient age was 0.75 years, and median ages of those infected by H . influenzae, N . meningitidis, and S . pneumoniae were 0.5, 4.0 and 0.6 years, respectively . The overall case fatality rate was 28% . The mortality rate of children below the age of 1 year was 38.4% and 13.8% for those above 1 year.

Ann Trop Paediatr, 1995 Sep, 15(3), 193 - 5
Type b Haemophilus influenzae endocarditis in children: case report and review of the literature; Malik AS; Haemophilus influenzae type b (Hib) endocarditis is a rare but potentially lethal condition . Only ten cases have been reported in the English literature . This report describes an 8-month-old Malay child with a ventricular septal defect who developed Hib endocarditis and died after 4 weeks of hospitalization . The literature is reviewed and previously reported cases summarized.

J Int Med Res, 1995 Sep-Oct, 23(5), 315 - 27
Lower respiratory tract infection therapy--the role of ciprofloxacin; Ball AP et al.; Lower respiratory tract infections account for a large proportion of prescribed antibiotics and, with emerging resistance to standard agents, the introduction of the fluoroquinolones, in particular ciprofloxacin, has provided a further component in the armamentarium . This review encompasses 37 published clinical trials which featured ciprofloxacin; 3274 patients with lower respiratory tract infections were treated with this agent; in 94.1% of patients treatment was clinically successful and 90.9% of cases showed eradication of the causative pathogen . When these data were supplemented with previously unpublished information from the clinical trial database, specific organism eradication rates of 86.1%, 96.2% and 94.6% for Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, respectively, were observed . These findings suggest that the high respiratory tissue penetration of ciprofloxacin and the achievable minimum inhibitory concentrations lead to acceptable clinical outcomes in lower respiratory tract infections.

Clin Infect Dis, 1995 Sep, 21(3), 639 - 42
Frequency of unrecognized Bordetella pertussis infections in adults; Deville JG et al.; To investigate the frequency of unrecognized Bordetella pertussis infections in adults, we performed IgA and IgG ELISA antibody studies with four B . pertussis antigens--i.e., lymphocytosis-promoting factor, filamentous hemagglutinin, pertactin, and fimbriae-2--in 51 health care workers from whom six consecutive yearly serum samples (from 1984 to 1989) were available . Overall, 90% of the subjects had a significant increase in antibody (IgA or IgG) to one or more antigens between 2 consecutive years during the 5-year study period; 55% of subjects had evidence of two infections, 17% had three infections, and 4% had four infections . Infections occurred in all study years, with the following rates: 1984-1985, 32%; 1985-1986, 24%; 1986-1987, 40%; 1987-1988, 29%; and 1988-1989, 43% (P = .12) . Some antibody rises may have been due to responses to cross-reacting antigens (Bordetella parapertussis, nontypable Haemophilus influenzae), but overall these data suggest that B . pertussis infections in adults are common, endemic, and usually unrecognized.

J Bacteriol, 1995 Sep, 177(18), 5316 - 21
Structural studies of lipooligosaccharides from Haemophilus ducreyi ITM 5535, ITM 3147, and a fresh clinical isolate, ACY1: evidence for intrastrain heterogeneity with the production of mutually exclusive sialylated or elongated glycoforms; Schweda EK et al.; The structures of the lipooligosaccharides (LOSs) from Haemophilus ducreyi ITM 5535 and ITM 3147 and a fresh clinical isolate, ACY1, have been investigated . Oligosaccharides were obtained from phenol-water-extracted LOS by mild acid hydrolysis and were studied by methylation analysis, fast atom bombardment and electrospray ionization mass spectrometry, and nuclear magnetic resonance spectroscopy . The major oligosaccharide obtained from all strains was a nonasaccharide with the structure beta-D-Galp-(1-->4)-beta-D-GlcNAcp-(1-->3)-beta-D-Galp-(1-->4)-D-a lpha-D-Hepp- (1-->6)-beta-D-Glcp-(1-->{L-alpha-D-Hepp-(1-->2)-L-alpha-D-Hepp - (1-->3)}4)-L-alpha-D-Hepp-Kdo (Kdo stands for 3-deoxy-D-manno-octulosonic acid) and is thus identical to that identified as the major oligosaccharide in H . ducreyi ITM 2665 (E . K . H . Schweda, A . C . Sundstrom, L . M . Eriksson, J.A . Jonasson, and A . A . Lindberg, J . Biol . Chem . 269:12040-12048, 1994) . Electrospray ionization mass spectrometry on O-deacylated LOS from H . ducreyi ITM 5535 obtained after treatment with anhydrous hydrazine gave evidence for the presence of a sialylated major compound, Neu5Ac alpha(2-->3)-beta-D-Galp-(1-->4)-beta-D-GlcNAcp-(1-->3)-beta-D-Gal p- (1-->4)-D-alpha-D-Hepp-(1-->6)-beta-D-Glcp-(1-->{L-alpha-D-Hepp -(1-->2)-L- alpha-D-Hepp-(1-->3)}4)-L-alpha-D-Hepp-Kdo(P)-O-deacylated lipid A (Neu5Ac stands for N-acetylneuraminic acid) . However, an even larger oligosaccharide could be isolated from all strains as a minor component, viz., the undecasaccharide beta-D-Galp-(1-->4)-beta-D-GlcNAcp-(1-->3)-beta-d-Galp-(1-->4)-beta-D-glcNAcp-(1-->3)-beta-D-Galp-(1-->4)-D-alpha-D-Hepp-(1-->6)-beta-D-Glcp-(1-->{L-alpha-D-Hepp-(1-->2)-L-alpha-D-Hepp-(1-->3)}4-L-alpha-D-Hepp-Kdo, which represents an N-acetyl lactosamine disaccharide unit elongation of the LOS outer core . No Sialylation of this latter minor component undecasaccharide was detected.

Pediatrics, 1995 Sep, 96(3 Pt 2), 576 - 9
Simultaneous administration of Haemophilus influenzae type b vaccine with acellular or whole-cell pertussis vaccine: effects on reactogenicity and immune responses to pertussis vaccines; Rennels MB et al.; OBJECTIVE . To evaluate the effect of simultaneous Haemophilus influenzae type b conjugate (Hib) vaccination on the safety and immunogenicity of selected acellular (DTaP) and whole-cell (DTP) pertussis vaccines with diphtheria and tetanus toxoids combined . METHODS . Enrollment of infants into a large multicenter study of the safety and immunogenicity of 13 DTaP and 2 DTP vaccines was partially completed when the first Hib vaccine, HbOC (Haemophilus b oligosaccharide conjugate vaccine), was licensed for use in infants . Thereafter, at each immunization most infants received HbOC simultaneously with DTaP (or DTP), administered in opposite thighs . Postvaccination geometric mean titers or concentrations (GMTs) of pertussis antibodies as measured by six different assays were compared pairwise among groups of infants receiving 0, 1, 2, or 3 simultaneous HbOC immunizations . The incidence of reactions was compared between infants who received only DTaP or DTP and those who received HbOC simultaneously . RESULTS . Comparison of postvaccination GMTs was possible among groups of infants receiving different numbers of simultaneous immunizations for 10 of the 13 DTaP and both DTP vaccines . Increased HbOC exposure had no consistent dose-response effect on antibody titers for DTaP or DTP vaccines in any assay . Significant differences between groups in postvaccination GMTs were observed with 4 DTaP vaccines in 1 to 2 assays each; the GMTs were higher with increasing HbOC exposure for 2 DTaP vaccines and lower for 2 others . There was no significant increase in reactions with simultaneous HbOC and DTaP immunization . CONCLUSIONS . Based on these retrospective analyses, there did not seem to be an interference in pertussis immunogenicity or alteration in reactogenicity associated with the simultaneous administration of HbOC and DTaP . These findings are encouraging with respect to the development of DTaP-Hib combination vaccines.

Arch Pediatr Adolesc Med, 1995 Sep, 149(9), 1001 - 8
Long-term outcome of Haemophilus influenzae meningitis in Navajo Indian children; D'Angio CT et al.; OBJECTIVES: To determine the long-term neurologic, cognitive, and educational outcomes of Navajo children who survived Haemophilus influenzae type b meningitis . DESIGN: Retrospective cohort study, with 3.6- to 15.0-year follow-up . SETTING: Navajo Indian reservation . PARTICIPANTS: Population-based cohort of 76 Navajo children with Haemophilus meningitis at less than 5 years of age between 1975 and 1986, with 41 (54%) consenting to undergo follow-up in 1990 . Each case was matched to one nearest-age sibling and one unrelated age-matched control . MAIN OUTCOME MEASURES: Standard intelligence test scores, neurologic abnormalities, and school performance . RESULTS: The mean IQ for cases was lower than that for siblings (79 vs 87, P = .006) or age-matched controls (79 vs 95, P < .001) . Twenty-nine percent of cases had severe neurologic sequelae, including mental retardation (24%), severe hearing loss (5%), cerebral palsy (7%), and seizure disorder (12%) . Eight percent of siblings (relative risk for cases vs siblings, 8.0; P = .05) and 2% of age-matched controls (relative risk vs cases, 10.0; P = .01) had mental retardation . No siblings or age-matched controls had any other severe neurologic sequela . Twenty-nine percent of cases, 23% of siblings (relative risk, 2.5; P = .45), and 0% of age-matched controls (P = .001) required special education services, while 42% of cases, 23% of siblings (relative risk, 3.3; P = .10), and 11% of age-matched controls (relative risk, 4.0; P = .005) had been retained in a grade in school . CONCLUSIONS: Navajo survivors of Haemophilus meningitis suffer more long-term neurologic, cognitive, and school-related disability than siblings or age-matched controls . They may also suffer higher morbidity than Haemophilus meningitis survivors in the general population.

Pediatrics, 1995 Sep, 96(3 Pt 1), 424 - 7
Epiglottitis and Haemophilus influenzae immunization: the Pittsburgh experience--a five-year review; Gonzalez Valdepena H et al.; OBJECTIVE . Current trends in the clinical presentation and management of children with epiglottitis at Children's Hospital of Pittsburgh were reviewed for the years 1988 to 1993 . METHODOLOGY . The medical records of all patients diagnosed as having epiglottitis between July 1988 and June 1993 at the Children's Hospital of Pittsburgh were reviewed . An additional telephone survey was conducted among the primary care physicians of those patients to collect information regarding administration of Haemophilus influenzae type b (HIB) vaccines . RESULTS . During the study period 28 children (age range, 11 months to 11 years, 10 months) were admitted with the diagnosis of epiglottitis . Cases declined remarkably in 1991 . Fever, sore throat, and stridor were the usual symptoms . HIB was the most common cause of epiglottitis accounting for 21 cases . Candida albicans was recovered from the surface culture of the epiglottis in two patients . At least 11 children experienced vaccine failure: nine with polysaccharide vaccine and two with the conjugate vaccine for HIB . CONCLUSION . Cases of epiglottitis have declined dramatically since licensure of HIB conjugate vaccines for use in early infancy . At least 52% of the reported cases represent vaccine failures with the purified polysaccharide vaccine.

J Med Microbiol, 1995 Sep, 43(3), 192 - 200
Localisation and immunological properties of a 24-kDa surface protein of Haemophilus ducreyi; Frisk A et al.; The cell wall and outer structures of Haemophilus ducreyi bacteria were investigated . The 24-kDa outer protein from two strains was purified with an SDS-PAGE preparative continuous-elution electrophoresis cell . The protein was further characterised by SDS-PAGE and immunoblotting, and the immunological properties were investigated by ELISA . Localisation on the bacterial surface was investigated by immuno-electron-microscopy with a polyclonal antiserum raised against the purified protein . A triple-laminar cell wall typical of gram-negative bacteria, close cellular contact between bacterial cells and outer blebs were seen on thin sections . An additional high mol . wt band of c . 165 kDa was seen when not treated by heating to 100 degrees C . A high density fibrilla-like material was detected on the bacterial cell and in the environment by negative staining and immuno-electron-microscopy with antisera specific for the 24-kDa protein . The surface localisation of the 24-kDa protein was confirmed by an ELISA technique with the specific antiserum and whole bacterial cells as antigen . The presence of antibodies to the 24-kDa protein was demonstrated in antisera to 13 strains of H . ducreyi, indicating antigenic identity or within-species cross-reactivity . Low titres of antibodies to this protein were also detected in 19 antisera raised against different strains of gram-negative bacteria, indicating cross-reactivity with other species . Antibody response to the 24-kDa protein in rabbits immunised subcutaneously with live bacteria resulted in a secondary IgG response . Of 28 sera from patients with culture-verified chancroid, 26 manifested high titres of IgG antibodies to the 24-kDa protein, thus indicating the involvement of this antigen in the disease process in man.

Infect Immun, 1995 Sep, 63(9), 3555 - 8
Role of the 145-kilodalton surface protein in virulence of the Brazilian purpuric fever clone of Haemophilus influenzae biogroup aegyptius for infant rats; Rubin LG; Brazilian purpuric fever (BPF) is a fulminant infection associated with bacteremia with clonally related strains of Haemophilus influenzae biogroup aegyptius . Case-associated clone strains are more virulent for infant rats than are non-BPF case-associated H . influenzae biogroup aegyptius isolates . I sought to determine the possible role of P145, a 145-kDa surface protein of BPF case H . influenzae biogroup aegyptius clone isolates, in virulence . First, I compared the virulence of two case-associated clone isolates from the blood of children with BPF from Serrana, Brazil, which differed in P145 expression but were identical in all other phenotypic and genotypic characteristics studied . Twenty-four hours after intraperitoneal inoculation of infant rats, there was a significantly higher incidence (51 versus 26%; P = 0.035) and magnitude (2.9 +/- 5.8 versus 0.7 +/- 2.0 CFU/0.01 ml; P = 0.024) of bacteremia in rats inoculated with the P145-expressing strain . I next compared the virulence of a P145-expressing case-associated clone strain with two P145-nonexpressing phase variants of this strain . One variant exhibited a lower mean magnitude of bacteremia and one displayed a similar magnitude of bacteremia compared with that displayed the P145-expressing parental strain . P145-expressing revertants of the P145-nonexpressing strains exhibited the same virulence as the P145-negative variants from which they were derived . Colonies grown from blood cultures maintained the P145 phenotype of the inoculated strain . These results suggest that P145 expression does not contribute to the virulence of the BPF case clone strain for infant rats following intraperitoneal inoculation.

Clin Pediatr (Phila), 1995 Sep, 34(9), 466 - 70
Factors associated with improved immunization rates for urban minority preschool children; Mitchell CK et al.; Urban, poor, preschool children are noted for having low immunization rates . To determine factors related to completion of immunization, vaccine records of 479 3-year-old children from an inner-city pediatric clinic were reviewed . Complete immunization was defined as four diphtheria-tetanus-pertussis doses, three oral polio vaccine doses, one measles-mumps-rubella dose, and one Haemophilus influenzae type b vaccine dose . Seventy percent of our patients were up-to-date by 2 years of age . The administration of all age-appropriate vaccines at a single visit for patients 15 months and older, the establishment of a continuous primary-care relationship, earlier age at first immunization, and lower birth weight were significantly associated with higher immunization levels in our study.

Microbiology, 1995 Sep, 141 ( Pt 9), 2271 - 9
Bacterial {Cu,Zn}-superoxide dismutase: phylogenetically distinct from the eukaryotic enzyme, and not so rare after all!
Kroll JS, Langford PR, Wilks KE, Keil AD.
Copper- and zinc-containing superoxide dismutases ({Cu,Zn}-SODs) are generally considered almost exclusively eukaryotic enzymes, protecting the cytosol and extracellular compartments of higher organisms from damage by oxygen free-radicals . The recent description of a few examples of bacterial forms of the enzyme, located in the periplasm of different Gram-negative micro-organisms, prompted a re-evaluation of this general perception . A PCR-based approach has been developed and used successfully to identify bacterial genes encoding {Cu,Zn}-SOD in a wide range of important human and animal pathogens-members of the Haemophilus, Actinobacillus and Pasteurella (HAP) group, and Neisseria meningitidis . Comparison of {Cu,Zn}-SOD peptide sequences found in Haemophilus ducreyi, Actinobacillus pleuropneumoniae, Actinobacillus actinomycetemcomitans, Pasteurella multocida, and N . meningitidis with previously described bacterial proteins and examples of eukaryotic {Cu,Zn}-SOD has shown that the bacterial proteins constitute a distinct family apparently widely separated in evolutionary terms from the eukaryotic examples . The widespread occurrence of {Cu,Zn}-SOD in the periplasm of bacterial pathogens, appropriately located to dismute exogenously derived superoxide radical anions, suggests that this enzyme may play a role in the interactive biology of organisms with their hosts and so contribute to their capacity to cause disease.

J Periodontal Res, 1995 Sep, 30(5), 325 - 31
Microflora in adult periodontitis; Petsios A et al.; The subgingival microflora of adult periodontitis was studied in 8 adults (36-47 years) and compared with that of 10 periodontally healthy individuals (36-43 years) . A total of 64 periodontal lesions were examined, and classified according to the attachment level in three categories: attachment loss > 6 mm, attachment loss 4-6 mm and attachment loss < 4 mm . Also for comparative purposes 20 gingival sulci were evaluated . Samples were taken using three standardized paper points and were incubated anaerobically in selective and non-selective media . The results showed a statistically significant association of Capnocytophaga gingivalis and Capnocytophaga sputigena with moderate periodontal lesions, while Haemophilus segnis has been correlated to severe periodontal lesions . We concluded that C . gingivalis, C . sputigena and H . segnis might be potentially conductive to periodontal deterioration in adult periodontitis.

J Clin Microbiol, 1995 Sep, 33(9), 2297 - 303
Simplified procedure for preparation of sensitized latex particles to detect capsular polysaccharides: application to typing and diagnosis of Actinobacillus pleuropneumoniae; Inzana TJ; A novel, inexpensive method for obtaining immunoglobulin G (IgG) specific for capsular antigen is described for use in latex agglutination tests . Hyperimmune rabbit serum against encapsulated Actinobacillus pleuropneumoniae was thoroughly adsorbed with a nonencapsulated mutant . The capsule titer of the absorbed serum was unaffected, whereas reactivity to nonencapsulated cells was reduced to background levels, as determined by enzyme immunoassay . The IgG component of the adsorbed serum was recovered by protein A chromatography and was covalently coupled through a water-soluble carbodiimide to carboxylate latex beads . The sensitized latex particles (SLP) were agglutinated by 10 ng of homologous capsule or more per ml, were not agglutinated by heterologous capsules at concentrations of < 10 micrograms/ml, and were stable for over 1 year at 4 degrees C without loss of sensitivity . There was no difference in the sensitivity or specificity of latex particles coupled with IgG purified by capsule affinity chromatography . The SLP were agglutinated by all strains of bacteria of the homologous serotype but not by heterologous serotypes or strains of Pasteurella multocida, Actinobacillus suis, or Haemophilus parasuis tested at a density equivalent to a 0.5 McFarland standard . The SLP detected homologous capsule in lung tissue, nasal swabs, and concentrated urine samples from all pigs culture positive for A . pleuropneumoniae but one . Precoating of carboxylate latex particles with avidin followed by conjugation of biotin-hydrazide-labelled IgG to capsule increased the sensitivity of the assay approximately 10-fold . Adsorption of serum with nonencapsulated mutants may be used to prepare SLP with optimum sensitivity and specificity without the need to purify capsule or couple capsule to affinity columns.

J Clin Microbiol, 1995 Sep, 33(9), 2254 - 9
Evaluation of mupirocin E-test for determination of isolate susceptibility: comparison with standard agar dilution techniques; Simpson IN et al.; Mupirocin E-test strips have been evaluated for their ease of use and accuracy in determining the susceptibilities of 171 strains of Staphylococcus spp., Streptococcus spp., Haemophilus influenzae, and Moraxella catarrhalis . The susceptibility of each strain was determined on two occasions, using parallel E-test and agar dilution methodologies each time . To ensure similar precisions for statistical analyses, E-test MICs were rounded up to a standard twofold agar dilution scale . Clear, elliptical zones were obtained against Staphylococcus spp . M . catarrhalis also gave clear zones, but the scale intercept was often difficult to interpret because of the irregular shape of the inhibition zone . Poor growth sometimes resulted in less-distinct zones of inhibition against Streptococcus spp . and H . influenzae . Excellent correlation was observed between the the E-test and agar dilution against Staphylococcus spp . and H . influenzae, with > 95% of the E-test values falling within one log2 dilution of the corresponding agar MIC . The correlation was lower for Streptococcus spp . and M . catarrhalis, with 86 and 83%, respectively, of E-test results falling within one log2 dilution of the agar MIC . When E-test MICs did not agree exactly with the corresponding agar MIC against Staphylococcus spp . or Streptococcus spp., there was a tendency for the E-test to give a lower MIC . This bias has little effect upon individual MICs in staphylococci or in the generation of susceptibility interpretation errors ( < 1.5% overall), but it could reduce population geometric mean MICs by factors of 0.78 to 0.83 . This effect was more marked for Streptococcus spp., reducing the population mean by a factor of 0.73 and resulting in 0.7% major and 8% very major errors . In contrast, the E-test tended to give higher MICs against M . catarrhalis, resulting in 7.3% major errors and increasing the population geometric mean MIC by a factor of 1.60.

J Clin Microbiol, 1995 Sep, 33(9), 2240 - 3
Comparison of in-house and commercially prepared haemophilus test media for disk diffusion testing of ampicillin against Haemophilus species; Laue RR et al.; The performance characteristics of two lots of Haemophilus Test Medium (HTM) prepared in-house at Mayo Clinic, one lot obtained from BBL (Cockeysville, Md.) and one lot obtained from Remel (Lenexa, Kans.), for disk diffusion susceptibility testing of ampicillin against 81 Haemophilus strains were assessed . Insufficient growth occurred in 2.5 to 13.6% of strains, depending on the HTM used, and was most frequent for beta-lactamase-negative, ampicillin-resistant strains when they were tested with one lot of HTM prepared in-house . Results for all 14 beta-lactamase-positive Haemophilus strains were in complete agreement for all lots of HTM when either 1990 or 1993 National Committee for Clinical Laboratory Standards (NCCLS) interpretive guidelines were followed . However, these beta-lactamase-positive Haemophilus strains could be as precisely, but more rapidly, identified by the cefinase disk method . Results for 56 beta-lactamase-negative, ampicillin-susceptible strains and 11 beta-lactamase-negative, ampicillin-resistant strains varied significantly among all HTM lots tested and as to whether 1990 or 1993 NCCLS interpretive guidelines were followed . We conclude that in our hands disk diffusion testing with HTM prepared in-house or obtained from two commercial sources and following either 1990 or 1993 NCCLS guidelines is an unacceptable method for determining the susceptibilities of Haemophilus strains to ampicillin.

Carbohydr Res, 1995 Aug 11, 272(2), 213 - 24
Structural studies of the saccharide part of the cell envelope lipooligosaccharide from Haemophilus influenzae strain galEgalK; Schweda EK et al.; The structure of the saccharide part of the lipooligosaccharide from Haemophilus influenzae strain galEgalK has been investigated . On treatment of the lipooligosaccharide with acid under mild conditions, followed by reduction with sodium borohydride and gel permeation chromatography, a main fraction was obtained which was studied by methylation analysis, NMR spectroscopy, and FABMS . The material was heterogeneous and contained two major compounds, A and B, and one minor, C . {formula: see text} In the structure, PEA is phosphoethanolamine, and L-D-Hep is L-glycero-D-manno-heptose . Kdo exists in reduced anhydro forms . The carbohydrate backbone is the same as that proposed for the saccharide part of the major component from H . influenzae type b strain A2 {N.J . Phillips, M . A . Apicella, J . M . Griffiss, and B . W . Gibson, Biochemistry, 32 (1993) 2003-2012}.

MMWR Morb Mortal Wkly Rep, 1995 Aug 4, 44(30), 567, 573 - 4
Chancroid detected by polymerase chain reaction--Jackson, Mississippi, 1994-1995; Upper respiratory tract infections - otitis media et al.; Department of Infectious Disease, Communicable Disease Centre, Tan Tock Seng Hospital, SingaporeManagement of the patient with otitis media, sinusitis or pharyngotonsillitis is based on information about the host, the organism and the antimicrobial agent . Otitis media (OM) is a common infection in children but selected children have recurrent and chronic OM . The predominant organisms responsible for OM are Streptococcus pneumonia, Haemophilus influenzae and Moraxella catarrhalis . Changes in the antimicrobial susceptibility govern the choice of antimicrobial agents . Surgical treatment should be considered if the child has persistent hearing loss in both ears . Sinusitis shares with OM similar pathogenesis, microbiology and choices of antimicrobial therapy . Endoscopic surgery is the treatment of choice for chronic sinusitis . Pharyngitis may be either viral or bacterial in origin . Penicillin remains the treatment of choice for bacterial pharyngotonsillitis . In patients with recurrent infection, the emergence of B-lactamase producing strains has to be considered and erythromycin or oral cephalosporins might be indicated.

Oral Microbiol Immunol, 1995 Aug, 10(4), 227 - 32
Characterization of tetracycline resistance in Actinobacillus actinomycetemcomitans; Roe DE et al.; Comparison of susceptibility data for Actinobacillus actinomycetemcomitans has been difficult because of the lack of standard susceptibility testing conditions . In this study, minimum inhibitory concentration to tetracycline was evaluated by comparing different media, air conditions and incubation times . Ten of 22 (45%) A . actinomycetemcomitans isolated from periodontally diseased sites grew on media supplemented with 4 micrograms per ml of tetracycline, but minimum inhibitory concentrations ranged from 0.125 to 8 micrograms/ml depending on the media and condition used . The best results were obtained with brain heart infusion agar (Difco Laboratories, Detroit MI) incubated in 5% CO2 for 48 h . Eighteen (82%) of the A . actinomycetemcomitans isolates hybridized with the Tet B determinant . The Tet B determinant was transferable between A . actinomycetemcomitans isolates as well as a Haemophilus influenzae recipient and appears to be associated with conjugative plasmids.

J Chemother, 1995 Aug, 7(4), 292 - 306
Bacterial pneumonia in adult patients with HIV infection; Moroni M et al.; Patients with HIV infection are at increased risk for community-acquired bacterial pneumonias, due in part to their defects in B-cell function . Streptococcus pneumoniae is the commonest cause of community-acquired pneumonia, with the second most common bacterial agent being Haemophilus influenzae . These two organisms account for about two-thirds of community-acquired bacterial pneumonias . Frequently bacterial pneumonias appear difficult to distinguish from Pneumocystis carinii pneumonia or other opportunistic lung infections, because of their atypical clinical and radiologic presentations . Community-acquired pneumonias may be recurrent but have low fatality rates . In comparison, nosocomial pneumonias occur primarily in patients with AIDS and are usually due to Staphylococcus aureus, Pseudomonas aeruginosa and other aerobic gram-negative bacilli . Nosocomial pneumonias have high fatality rates . S.aureus is an important cause of morbidity and mortality in patients with AIDS and has emerged as a secondary opportunist in lungs of patients with opportunistic diseases . While appropriate laboratory study is being done, empiric antibiotic therapy should be directed against the microorganisms above described.

Clin Infect Dis, 1995 Aug, 21(2), 409 - 14
Comparison of azithromycin and ceftriaxone for the treatment of chancroid; Martin DH et al.; We conducted a randomized, unblinded, prospective study designed to determine the efficacy of single-dose azithromycin for the treatment of chancroid . Men and women 16 years of age and older who had darkfield-negative genital ulcers that were clinically suspected to be caused by Haemophilus ducreyi and who attended urban sexually transmitted disease clinics or presented to hospital emergency departments were enrolled in the study . Patients were randomized to receive 250 mg of ceftriaxone im or 1 g of azithromycin orally, both given as a single dose . They were followed for up to 23 days after treatment . For 65 patients, cultures were positive for H . ducreyi; there were 68 patients whose cultures were negative for both H . ducreyi and herpes simplex virus and who had no evidence of syphilis . All 133 patients returned for at least one follow-up visit . At the time of the last follow-up visit, all 32 patients whose cultures were positive for H . ducreyi and who were treated with azithromycin were clinically cured . In all 33 culture-positive cases in which ceftriaxone was used, there was either clinical improvement or cure at the time of the patient's last follow-up visit . In addition, azithromycin and ceftriaxone were equally effective in healing ulcers for which cultures were negative . We conclude that a single 1-g oral dose of azithromycin is as effective as a 250-mg im dose of ceftriaxone for the treatment of chancroid.

Mol Microbiol, 1995 Aug, 17(3), 555 - 64
Identification and characterization of a cell envelope protein of Haemophilus influenzae contributing to phase variation in colony opacity and nasopharyngeal colonization; Weiser JN et al.; Haemophilus influenzae undergoes spontaneous phase variation in colony morphology . Organisms from transparent colonies efficiently colonize the nasopharynx in an infant rat model of H . influenzae carriage, whereas organisms from more opaque colonies are deficient at colonization . A genetic approach relying on the transformability of H . influenzae was used to identify a locus contributing to opacity variation . By screening a library of chomosomal DNA from an opaque variant of strain Rd, it was possible to isolate a single clone capable of transforming a transparent Rd host to a more opaque phenotype . A region containing two genes, designated oapA and oapB, was identified . The deduced amino acid sequence of oapB has similarity to a consensus sequence for bacterial lipoproteins . Genetically defined mutations in oapA were transformed into the transparent Rd to confirm that this gene is required for expression of the transparent colony phenotype . Although oapA lacks a signal sequence, gene fusions to phoA show that OapA is secreted in H . influenzae and undergoes phase variation in expression . Mutagenesis of oapA in strain Rd, and type b strain Eagan, resulted in loss of the ability to colonize the nasopharynx of infant rats . The type b mutant, however, was as virulent as its parent strain when inoculated intraperitoneally . This suggests that the contribution of OapA to pathogenesis is limited to events associated with colonization of the mucosal surface.

J Otolaryngol, 1995 Aug, 24(4), 255 - 7
Unilateral supraglottitis in adults: fact or fiction; McQueen CT et al.; Supraglottitis and epiglottitis have been described for many years by various authors . Haemophilus influenzae type b is the primary cause of childhood epiglottitis, which classically appears between the ages of 2 and 4 years . Onset is usually acute and the presentation can be dramatic with drooling, high temperatures, and stridor . Compared to childhood supraglottitis, adult supraglottitis usually pursues a more indolent course with no significant airway compromise and no identifiable pathogen . Rarely, adult supraglottitis can resemble its childhood counterpart with acute respiratory compromise secondary to H . influenzae infection . Although most incidences of adult supraglottitis are infectious in origin and involve the entire supraglottitis and epiglottis, we present two cases of unilateral supraglottitis caused by inhalation of a hot wire screen used as a filter for smoking crack cocaine.

Clin Infect Dis, 1995 Aug, 21 Suppl 1, S77 - 83
Preventing bacterial respiratory tract infections among persons infected with human immunodeficiency virus; Keller DW et al.; Bacterial respiratory tract infections occur frequently in persons infected with human immunodeficiency virus (HIV) and may be caused by a wide variety of pathogens . Pneumonia is the most commonly diagnosed respiratory infection in HIV-infected persons and is more common in those persons than in non-HIV-infected ones . HIV-infected persons have a much higher risk of pneumococcal disease than do noninfected controls, and disease may occur relatively early in the course of HIV infection . While mortality associated with the disease does not seem to be high among HIV-infected persons, there is a higher rate of recurrence of the disease in that population . Risk factors for pneumococcal disease in HIV-infected persons are not well characterized . Though efficacy data are limited, the 23-valent polysaccharide pneumococcal vaccine is recommended for use early in the course of HIV infection . There are no data suggesting that HIV-infected persons should be revaccinated routinely . Antiretroviral agents may enhance the immunologic response to the polysaccharide vaccine . Prophylactic antibiotics may have a role in preventing recurrences of severe bacterial respiratory infections, and intravenous immunoglobulin may be useful in preventing serious bacterial infections in HIV-infected children . HIV-infected persons are also at greater risk for serious infections with Haemophilus influenzae than are non-HIV-infected persons . Vaccination against H . influenzae type b (Hib) is recommended for HIV-infected children but not for adults . Antimicrobial drug-resistant strains of Streptococcus pneumoniae and H . influenzae have become more prevalent recently and consequently have impacted on strategies for prevention and treatment of those infections.

Trends Microbiol, 1995 Aug, 3(8), 304 - 9
Phenotypic variation of carbohydrate surface antigens and the pathogenesis of Haemophilus influenzae infections; Roche RJ et al.; Phenotypic variation of two major carbohydrate surface antigens of Haemophilus influenzae, the capsule and lipopolysaccharide, exemplifies some of the genetic mechanisms used by pathogenic bacteria in interacting with host microenvironments . The ability to generate phenotypic variety at high frequency within clonal populations of microorganisms provides an adaptive mechanism to combat the polymorphisms and immune repertoires of the host.

Epidemiol Infect, 1995 Aug, 115(1), 89 - 100
Epidemiology of invasive Haemophilus influenzae infections in England and Wales in the pre-vaccination era (1990-2); Anderson EC et al.; This survey defined the pattern of invasive Haemophilus influenzae infections during 1990-2 in six regions in England and Wales during the pre-vaccination era providing a baseline against which any changes in patterns of disease due to the introduction of the Haemophilus influenzae type b vaccination programme can be monitored . A total of 946 cases of invasive Haemophilus influenzae were recorded during the survey period of which almost 90% were due to type b and most of the remainder were non-typeable . Type b infections occurred predominantly in children less than 5 years of age (88%) with the highest attack rate in male infants in the 6-11 month age group . Diagnostic category varied with both age and serotype; meningitis was the commonest presentation overall but pneumonia and bacteraemia were more common in adults and non-typeable isolates . Mortality was highest in neonates and the elderly (over 65 years of age) who were more likely to have an underlying predisposing condition than older children and adults . Children under 5 years of age had a higher case fatality rate for non-typeable than for type b infections . Ampicillin resistance was 15% and there were no cefotaxime resistant type b isolates.

Ann Otol Rhinol Laryngol, 1995 Aug, 104(8), 646 - 52
Quantitative bacteriology of tonsils removed from children with tonsillitis hypertrophy and recurrent tonsillitis with and without hypertrophy; Kuhn JJ et al.; The aerobic and anaerobic bacterial species and their numbers were studied in tonsillar specimens from children who had undergone elective tonsillectomy: 6 patients with recurrent tonsillitis (RT), 9 with recurrent tonsillitis with hypertrophy (RTH), and 8 with obstructive tonsillar hypertrophy (OTH) . Mixed flora were present in all tonsils, yielding an average of 6.7 isolates (5.6 aerobic or facultative and 1.1 anaerobic bacteria) . The highest recovery rate of organisms per tonsil was in patients with OTH (7.7 per tonsil), compared to 6.3 per tonsil in RT and 5.9 per tonsil in RTH . The predominant aerobic and facultative organisms were Haemophilus influenzae (22 isolates), Neisseria sp (16), Staphylococcus aureus (14), and Eikenella corrodens (14), and the predominant anaerobic bacteria were Fusobacterium sp (8), Bacteroides sp (7), and Prevotella melaninogenica (5) . The number of bacteria per gram of tonsillar tissue varied between 10(4) and 10(8) . A higher concentration of S aureus and H influenzae was found in hypertrophic tonsils (RTH and OTH) as compared to RT . These findings suggest the presence of an increased bacterial load and supports an etiologic role for H influenzae and S aureus in hypertrophic tonsils with and without inflammation (RTH and OTH) . Further studies to elucidate the effect of selective antimicrobial therapy directed at these organisms may offer an alternative management of hypertrophic tonsils.

South Med J, 1995 Aug, 88(8), 797 - 804
Antibiotic resistance in bacteria; Berkowitz FE; Antibiotic resistance in bacteria has emerged as a medical catastrophe . This results from the speed at which bacteria multiply and are spread, and the ease with which they can change their genetic material or acquire new genes . They exert biochemical resistance by preventing entry of the drug, by rapidly extruding the drug, or by enzymatically inactivating the drug or altering its molecular target . The presence of antibiotics in the internal environments of human beings and animals provides a selective pressure for any resistant organisms to become predominant . Examples of antibiotic resistance in several important human pathogens are Streptococcus pneumoniae, enterococci, staphylococci, enteric bacilli, Haemophilus influenzae, Neisseria gonorrhoeae, Neisseria meningitidis, and Mycobacterium tuberculosis.

Pediatrics, 1995 Aug, 96(2 Pt 1), 216 - 9
Antibody response of low birth weight infants to Haemophilus influenzae type b polyribosylribitol phosphate-outer membrane protein conjugate vaccine; Munoz A et al.; OBJECTIVE . To evaluate the effectiveness in low birth weight (LBW) infants of the currently recommended immunization schedule for conjugated Haemophilus influenzae type b (HIB) vaccine . METHODS . We quantified antibody responses in 36 preterm infants with a mean birth weight of 1060 g and a mean gestational age of 28 weeks . Infants were immunized with 0.5 mL of HIB vaccine at 2 and 4 months' postnatal age . Specific HIB antibodies were quantified on cord blood, immediately before each immunization and 2 months after the last immunization . RESULTS . Even though the geometric mean titers increased significantly during the study period, they were still markedly lower than values reported in term infants . After the second immunization, only 24 infants (67%) attained antibody concentrations of more than 0.25 micrograms/mL, defined as seropositivity . Also, only 53% of infants achieved antibody concentrations of more than 1.0 micrograms/mL compared with 92% as reported in term infants . Stepwise logistic regression identified gestational age of 27 weeks or less and the amount of intravenous immunoglobulin received as the significant variables influencing the antibody response after the first immunization . The incidence of side effects was negligible . CONCLUSIONS . We conclude that LBW infants, and especially those born at 27 or less weeks' gestation, do not respond as effectively to the HIB vaccine . We speculate that reevaluation of the current immunization schedule may be required for very LBW infants.

J Med Microbiol, 1995 Aug, 43(2), 120 - 4
The elucidation of novel capsular genotypes of Haemophilus influenzae type b with the polymerase chain reaction; Leaves NI et al.; Molecular characterization is an important pre-requisite for post-vaccine studies of Haemophilus influenzae type b (Hib) . Three capsular genotyping patterns, b(S), b(G) and b(V), have been described in the major phylogenetic lineage of Hib . However, in a recent series of prospective studies, three new hybridisation patterns were observed among 425 strains of Hib . Four pairs of polymerase chain reaction (PCR) primers were used to identify the capsular gene (cap) structure of these Hib strains . This showed that the strains possessed simple DNA re-arrangements . In two instances a change in restriction enzyme recognition site was the most likely cause of the new hybridisation pattern . The third strain possessed a cap b locus consisting of intact tandem repeats of cap b in a b(S) background . It was reasoned that a similar cap b locus would not be readily recognised by hybridisation in a b(G) background, and b(G) strains were therefore characterized by the PCR method . This showed one of 35 b(G) strains to possess a cap locus with intact tandem repeat copies of cap b . The novel capsular genotypes described here are rare, but can be detected rapidly and accurately by a combination of PCR and capsular genotyping hybridisation patterns.

J Infect Dis, 1995 Aug, 172(2), 446 - 52
Chancroid epidemiology in New Orleans men; DiCarlo RP et al.; Epidemiologic, clinical, and microbiologic data were collected from 299 men with nonsyphilitic genital ulcer disease . One hundred eighteen (39%) were culture-positive for Haemophilus ducreyi, 57 (19%) were culture-positive for herpes simplex virus, and 124 (41%) were culture-negative . Patients with chancroid were significantly more likely than those with genital herpes to have been frequent users of alcohol (44% vs . 23%, P = .006) . They were also more likely recently to have used cocaine (25% vs . 9%, P = .013), had sex with a prostitute (17% vs . 5%, P = .035), traded drugs for sex (16% vs . 2%, P = .005), and had a sex partner who used drugs (38% vs . 13%, P = .001) . Culture-negative patients were similar to chancroid patients with respect to most epidemiologic risk factors . Despite the epidemiologic similarities, the clinical features of culture-negative ulcers resembled those of culture-proven herpes ulcers more closely than they did those of culture-proven chancroid ulcers . These data establish a link between chancroid in the United States and the use of crack cocaine.

Infect Immun, 1995 Aug, 63(8), 3094 - 100
Swine model of Haemophilus ducreyi infection; Hobbs MM et al.; Haemophilus ducreyi is a strict human pathogen that causes sexually transmitted genital ulcer disease . We infected domestic swine with H . ducreyi 35000, resulting in the development of cutaneous ulcers histologically resembling human chancroid lesions . Intraepidermal lesions progressed from pustules to ulcers containing polymorphonuclear leukocytes and were accompanied by a dermal inflammatory infiltrate containing T cells and macrophages . H . ducreyi was recovered from lesions up to 17 days after inoculation, and pigs did not develop immunity to reinfection with the challenge strain . Features of the model include inoculation through abrasions in the epidermis, ambient housing temperatures for infected pigs, the ability to deliver multiple different inocula to a single host, and the availability of monoclonal antibodies against porcine immune cells permitting immunohistochemical characterization of the host immune response to H . ducreyi infection.

Infect Immun, 1995 Aug, 63(8), 3054 - 61
The outer membranes of Brucella spp . are resistant to bactericidal cationic peptides; Martinez de Tejada G et al.; The actions of polymyxin B, rabbit polymorphonuclear lysosome extracts, 14 polycationic peptides (including defensin NP-2, cecropin P1, lactoferricin B, and active peptides from cationic protein 18 and bactenecin), EDTA, and Tris on Brucella spp . were studied, with other gram-negative bacteria as controls . Brucella spp . were comparatively resistant to all of the agents listed above and bound less polymyxin B, and their outer membranes (OMs) were neither morphologically altered nor permeabilized to lysozyme by polymyxin B concentrations, although both effects were observed for controls . EDTA and peptides increased or accelerated the partition of the hydrophobic probe N-phenyl-naphthylamine into Escherichia coli and Haemophilus influenzae OMs but had no effect on Brucella OMs . Since Brucella and H . influenzae OMs are permeable to hydrophobic compounds (G . Martinez de Tejada and I . Moriyon, J . Bacteriol . 175:5273-5275, 1993), the results show that such unusual permeability is not necessarily related to resistance to polycations . Although rough (R) B . abortus and B . ovis were more resistant than the controls were, there were qualitative and quantitative differences with smooth (S) brucellae; this may explain known host range and virulence differences . Brucella S-lipopolysaccharides (LPSs) had reduced affinities for polycations, and insertion of Brucella and Salmonella montevideo S-LPSs into the OM of a Brucella R-LPS mutant increased and decreased, respectively, its resistance to cationic peptides . The results show that the core lipid A of Brucella LPS plays a major role in polycation resistance and that O-chain density also contributes significantly . It is proposed that the features described above contribute to Brucella resistance to the oxygen-independent systems of phagocytes.

Infect Immun, 1995 Aug, 63(8), 2989 - 94
Functional capacities of clonal antibodies to Haemophilus influenzae type b polysaccharide; Nahm MH et al.; Haemophilus influenzae type b (Hib) is an important pathogen for young children, and children can be protected with antibodies (Abs) to Hib polysaccharide (PS) capsule, a linear polymer of ribosyl ribitol phosphate . The structure of anti-Hib-PS Abs has been well characterized at the molecular level; about two-thirds of anti-Hib-PS Abs use a V kappa gene named A2, and the remaining anti-Hib-PS Abs use one of many other VL genes . In order to understand the structural basis for the variability in the function of these Abs, we prepared 18 clonally pure Abs from adults and studied their affinity, avidity, bactericidal potency in vitro, and ability to reduce bacteremia in newborn rats . Affinities and avidities were determined as the inverse of the concentrations of short (3 repeating units) and long (20 repeating units) ligands which could bind 50% of anti-Hib-PS Ab in solution, respectively . No significant correlations between the protection of newborn rats and affinity (r = 0.02) or avidity (r = 0.16) were observed . The amount of Ab required to kill 50% of bacteria in vitro decreased with avidity (r = -0.32), as expected . However, Abs with high affinity were unexpectedly found to have less bactericidal activity (r = 0.38) . This suggests that avidity may be a better predictor of Ab function than affinity . Affinity and avidity results were negatively correlated (r = 0.76, P = 0.0022), and Abs that had A2 V kappa gene products had higher avidity (P < 0.05) and lower affinity (P = 0.06) than Abs that had other VL genes . A possible explanation of these observations is that the epitope for Abs with the A2 gene is within the Hib-PS chain itself, whereas the epitope for Abs with a non-A2 gene is the terminus of Hib-PS.

Infect Immun, 1995 Aug, 63(8), 2976 - 82
A system for generalized mutagenesis of Haemophilus ducreyi; Stevens MK et al.; The lack of a generalized mutagenesis system for Haemophilus ducreyi has hampered efforts to identify virulence factors expressed by this sexually transmitted pathogen . To address this issue, the transposable element Tn1545-delta 3, which encodes resistance to kanamycin, was evaluated for its ability to insert randomly into the H . ducreyi chromosome and produce stable, isogenic mutants . Electroporation of H . ducreyi with 1 microgram of plasmid pMS1 carrying Tn1545-delta 3 resulted in the production of 10(4) kanamycin-resistant transformants; Southern blot analysis of a number of these transformants indicated that insertion of the transposon into the chromosome occurred at a number of different sites . This pMS1-based transposon delivery system was used to produce an H . ducreyi mutant that expressed an altered lipooligosaccharide (LOS) . Passage of this mutant in vitro in the presence or absence of kanamycin did not affect the stability of the transposon insertion . To confirm that the observed mutant phenotype was the result of the transposon insertion, a chromosomal fragment containing Tn1545-delta 3 was cloned from this H . ducreyi LOS mutant . Electroporation of the wild-type H . ducreyi strain with this DNA fragment yielded numerous kanamycin-resistant transformants, the majority of which had the same altered LOS phenotype as the original mutant . Southern blot analysis confirmed the occurrence of proper allelic exchange in the LOS-deficient transformants obtained in this backcross experiment . The ability of Tn1545-delta 3 to produce insertion mutations in H . ducreyi should facilitate genetic analysis of this pathogen.

Infect Immun, 1995 Aug, 63(8), 2931 - 40
Enhanced respiratory clearance of nontypeable Haemophilus influenzae following mucosal immunization with P6 in a rat model; Kyd JM et al.; Nontypeable Haemophilus influenzae (NTHi) is a common cause of infection of the respiratory tract in children and adults . The search for an effective vaccine against this pathogen has focused on components of the outer membrane, and peptidoglycan-associated lipoprotein P6 is among the proposed candidates . This study investigated the immunogenicity of P6 in a rat respiratory model . P6 was purified from two strains of NTHi, one capsule-deficient strain and an H . influenzae type b strain, and assessed for clearance of both homologous and heterologous bacterial strains following mucosal immunization . A protective immune response was determined by enhancement of pulmonary clearance of live bacteria and an increased rate of recruitment of phagocytic cells to the lungs . This was most effective when Peyer's patch immunization was accompanied by an intratracheal (IT) boost . However, the rate of bacterial clearance varied between strains, which suggests some differences in anti-P6 immunological defenses recognizing the expression of the highly conserved P6 lipoprotein on the bacterial surface in some strains . P6-specific antibodies in both serum and bronchoalveolar lavage fluid were cross-reactive and did not differ significantly in strain specificity, demonstrating that difference in clearance was unlikely due to differences in P6-specific antibody levels . Serum homologous and heterologous P6-antibody was bactericidal against NTHi even when enhanced clearance had not been observed . Peyer's patch immunization induced P6-specific CD4+ T-helper cell proliferation in lymphocytes isolated from the mesenteric lymph nodes . An IT boost increased the level of P6-specific antibodies in serum and bronchoalveolar lavage fluid, and P6-specific mesenteric node lymphocyte proliferation . Cells from rats immunized with P6 demonstrated proliferation following stimulation with P6 from nonhomologous strains; however, there was some variation in proliferative responses to P6 from different strains in lymphocytes isolated from animals immunized with killed bacteria . The increase in P6-specific antibodies and T-helper cell responses following an IT boost correlated with an increased rate of recruitment of phagocytic cells and enhanced bacterial clearance of both homologous and heterologous bacteria in the lungs . The data suggests that P6 has the potential to afford protection against pulmonary infection by NTHi following the induction of effective antigen-specific B- and T-cell responses in mucosal tissues.

Arch Otolaryngol Head Neck Surg, 1995 Aug, 121(8), 898 - 902
Acute epiglottitis and infant conjugate Haemophilus influenzae type b vaccination in northern Finland; Alho OP et al.; OBJECTIVE: To determine the incidence and characteristics of acute epiglottitis among children (< or = 19 years of age) and adults (> or = 20 years of age) before and after widespread conjugate Haemophilus influenzae type b vaccination for infants . DESIGN: A retrospective population-based survey over a 27-year period from 1967 through 1993 in 35 communities in a northern province of Finland with a population of approximately 300,000 . SETTING: An academic tertiary referral center . MAIN OUTCOME MEASURES: All acute epiglottitis cases in the area identified from the hospital discharge register and the regional autopsy register . RESULTS: The average incidence rate for children was 1.8 cases per 100,000 individuals per year (95% confidence interval {CI}, 1.3 to 2.5) . As no vaccine failures emerged, the incidence rate for children aged 0 to 4 years declined sharply once the vaccination started in 1986 from 7.6 (95% CI, 5.3 to 10.4) to 0 (95% CI, 0 to 3.3) cases per 100,000 individuals per year . By contrast, a fourfold increase in adult acute epiglottitis (incidence rate ratio, 4.6; 95% CI, 2.7 to 7.9) was detected after vaccination of the children, the average incidence rate for the whole period being 1.0 cases per 100,000 individuals per year (95% CI, 0.8 to 1.3) . No marked change in the adult patient profile was found during this increase, however . CONCLUSION: Acute epiglottitis practically vanished among young children in this population after conjugate H influenzae vaccination, but adult cases increased, the patient profile remaining the same.

Genitourin Med, 1995 Aug, 71(4), 224 - 7
Asymptomatic carriage of Haemophilus ducreyi confirmed by the polymerase chain reaction; Hawkes S et al.; OBJECTIVE--To investigate asymptomatic carriage of Haemophilus ducreyi by using polymerase chain reaction (PCR) on samples from women at high risk of infection . SUBJECTS--213 commercial sex workers (CSWs) recruited in The Gambia, West Africa . METHODS--Genital samples (cervical, vaginal and ulcer) were tested for the presence of H ducreyi by PCR with the technique of "one tube nested primer" . RESULTS--12 CSWs were PCR positive for H ducreyi; 8 of these women had genital ulcers on examination . CONCLUSION--Using a simplified PCR technique for detecting H ducreyi we have shown that 2% of CSWs were carrying the organism without clinical symptoms or signs . This has important implications for sexually transmitted disease control programmes in areas with a high prevalence of chancroid.

Arch Dis Child, 1995 Aug, 73(2), 160 - 1
Latex agglutination testing in bacterial meningitis; Finlay FO et al.; The value of the latex agglutination test in meningitis was assessed . This was positive in 60% cases of Streptococcus pneumoniae, 93% of Haemophilus influenzae type b, and 39% of Neisseria meningitidis infections . We cannot support the view that this test was more valuable than Gram staining in partially treated meningitis and cannot recommend its routine use . It may, however, be valuable if Gram staining does not identify an organism or if it suggests meningococcal infection.

J Clin Microbiol, 1995 Aug, 33(8), 2027 - 31
Turnover of nonencapsulated Haemophilus influenzae in the nasopharynges of otitis-prone children; Samuelson A et al.; Restriction enzyme analysis of total genomic DNA was applied to study the epidemiology of nontypeable Haemophilus influenzae (NTHI) isolated from the nasopharynges of children with recurrent acute otitis media (AOM) . The turnover of strains, as judged from genetic fingerprinting of a total of 213 H . influenzae isolates collected prospectively during a 2-year study period from 38 children under 3 years of age, was examined in relation to episodes of AOM as well as to courses of antibiotic treatment . The children were selected if they had had at least one episode of AOM before 1 year of age and if more than two nasopharyngeal isolates of H . influenzae were recovered . The 213 H . influenzae isolates (90% NTHI) recovered corresponded to 128 different DNA fingerprints . Fifty-eight percent of the fingerprints were observed only once, whereas 42% appeared on two or more occasions in isolates from the same individual or in close relatives, i.e., brothers and sisters . Sixty-seven percent of these strains had a minimum colonization period of 2 months or less . Intermittent nasopharyngeal colonization periods longer than 5 months could be demonstrated for 13% of the strains . The present data suggest that intermittent colonization is due to endogenous reinfections . Genetically identical NTHI strains from unrelated individuals were never identified . As expected from the observation of a relatively high proportion of persistent colonizations, no correlation was found between episodes of AOM and the acquisition of new strains of H . influenzae, nor was any direct relation between antimicrobial therapy and the elimination of nasopharyngeal colonization with a particular strain of H . influenzae observed.

Int J Pediatr Otorhinolaryngol, 1995 Aug, 33(1), 1 - 16
Otitis media in a population of black American and white American infants, 0-2 years of age; Casselbrant ML et al.; To determine the incidence of otitis media (OM) and the bacteriology of acute otitis media (AOM) in a clinic population of young children in Pittsburgh, 138 black infants and 60 white infants were followed from birth to 2 years of age, examined at monthly intervals and whenever an upper respiratory tract infection (URI) or OM intervened . By 24 months of age the cumulative incidence of episodes of AOM was 43% and 42%, and of episodes of middle-ear effusion (MEE) was 86% and 85% in black and white infants, respectively . The average rate of episodes of AOM was 0.41 and 0.39 and of episodes of MEE was 1.68 and 1.70 in black and white infants, respectively . Tympanocentesis was performed for episodes of AOM and the following organisms were isolated from black and white infants, respectively: Streptococcus pneumoniae 43% and 43% of episodes; Moraxella catarrhalis 24% and 24%; non-typable Haemophilus influenzae 18% and 24%; and Haemophilus influenzae type b 5% and 0% . In both black and white infants first born children had less ear disease . We found no difference in the incidence of otitis media during the first 2 years of life between black and white infants.

Arch Oral Biol, 1995 Aug, 40(8), 753 - 63
Bacteria in human mouths involved in the production and utilization of hydrogen peroxide; Ryan CS et al.; Earlier studies have demonstrated that pure cultures of oral streptococci produce hydrogen peroxide but none has found any free peroxide in dental plaque or salivary sediment despite streptococci being major components of their mixed bacterial populations . The absence of peroxide in plaque and sediment could be due to the dominance of its destruction over its formation by bacterial constituents . To identify which of the oral bacteria might be involved in such a possibility, pure cultures of 27 different oral bacteria were surveyed (as well as dental plaque and sediment) for their peroxide-forming and peroxide-removing capabilities . Peroxide production was measured for each of the pure cultures by incubation with glucose at low and high substrate concentrations (2.8 and 28.0 mM) for 4 h and with the pH kept at 7.0 by a pH-stat . Removal of hydrogen peroxide was assessed in similar experiments where peroxide at 0, 29.4, 147.2 or 294.4 mM {0, 0.1, 0.5 and 1% (w/v)} replaced the glucose . Hydrogen peroxide formation was seen with only three of the bacteria tested, Streptococcus sanguis I and II (sanguis and oralis), and Strep . mitior (mitis biotype I); levels of hydrogen peroxide between 2.2 and 9.8 mM were produced when these micro-organisms were grown aerobically and 1.1 and 3.9 mM when grown anaerobically . Earlier reports indicate that such levels were usually sufficient to inhibit the growth of many plaque bacteria . The amounts formed were similar at the two glucose levels tested, suggesting that maximum peroxide production is reached at low glucose concentration . None of the three peroxide-producing organisms was able to utilize hydrogen peroxide but five of the other 24 tested, Neisseria sicca, Haemophilus segnis, H . parainfluenzae, Actinomyces viscosus and Staphylococcus epidermidis, could readily do so, as could the mixed bacteria in salivary sediment and dental plaque, both of which contain relatively high numbers of these peroxide-utilizing micro-organisms . The ability of the bacteria in plaque and sediment to degrade hydrogen peroxide was considerable and extremely rapid; peroxide removal and usually complete within the first 15 min of the incubation even when its initial level was as high as 294.4 mM . This almost overwhelming ability to remove peroxide was confirmed when peroxide-producing and -using cultures were mixed and when each of eight salivary sediments was incubated with glucose and with peroxide at concentrations up to 294.4 mM . In the glucose incubations, no hydrogen peroxide was observed, indicating dominance of microbial peroxide removers over hydrogen peroxide producers.(ABSTRACT TRUNCATED AT 400 WORDS)

Antimicrob Agents Chemother, 1995 Aug, 39(8), 1820 - 3
Effect of xylitol on growth of nasopharyngeal bacteria in vitro; Kontiokari T et al.; Xylitol is known to reduce caries by inhibiting the growth of Streptococcus mutans . We hypothesized that xylitol could also affect the growth of other nasopharyngeal bacterial flora, which could be important when considering respiratory infections caused by these bacteria . We studied this in vitro by adding xylitol to the medium and observed that 1 and 5% xylitol reduced markedly the growth of alpha-hemolytic streptococci, including S . pneumoniae . It reduced slightly the growth of beta-hemolytic streptococci but not that of Haemophilus influenzae or Moraxella catarrhalis . The inhibitory growth pattern was similar to that previously seen with S . mutans, which may indicate a similarity in the enzymatic processing of five-carbon sugars such as xylitol . This sugar alcohol is a widely used sweetener, and the concentrations used in our experiments are easily achieved in the oral cavity . If xylitol reduces the growth of S . pneumoniae in the nasopharynx, it could also reduce the carriage of this pathogen and thus have clinical significance in the prevention of pneumococcal diseases.

Pediatr Res, 1995 Aug, 38(2), 182 - 6
Increased proportions of bacteria capable of cleaving IgA1 in the pharynx of infants with atopic disease; Kilian M et al.; Based on the observation that children with a history of atopic disease show significantly increased levels of cleaved secretory IgA in nasopharyngeal secretions, we have previously formulated the hypothesis that bacteria-induced local deficiencies of the immune barrier of the upper respiratory tract may be a contributing factor in the development and perpetuation of atopic diseases . To evaluate this hypothesis, 25 infants were subjected to clinical, bacteriologic, and immunologic examination at the age of 18 mo, 30 mo, and 5 y . The 11 infants, who showed clinical and immunologic evidence of atopic disease at the age of 18 mo, harbored significantly higher proportions of IgA1 protease-producing bacteria (median, 36%; range, 14-64%) than the 14 healthy infants (median, 5%; range, 0.4-14%) . No statistically significant differences were observed at the two subsequent examinations, but healthy children showed a statistically significant increase in proportions of IgA1 protease-producing bacteria in the pharynx with increasing age . IgA1 protease-producing bacteria detected included Streptococcus mitis biovar 1, Haemophilus influenzae, Haemophilus parahaemolyticus, Streptococcus pneumoniae, and Neisseria meningitidis, of which the first mentioned species was mainly responsible for the difference observed at the 18-mo examination . Percentage proportions of IgA1 protease-producing bacteria were significantly related to passive smoking which may stimulate the premature and more pronounced pharyngeal colonization of the atopic infants with the IgA1 protease-producing variant of S . mitis biovar 1 . The results of the study support the hypothesis that IgA1 protease-producing bacteria colonizing the upper respiratory tract jeopardize the local immune barrier and, thereby, may facilitate the penetration of potential allergens resulting in atopic disease.

Jpn J Antibiot, 1995 Aug, 48(8), 965 - 98
{Susceptibilities of bacteria isolated from patients with respiratory infectious diseases to antibiotics (1991)}; Ikemoto H et al.; Isolated bacteria from respiratory tract infections were collected since 1981 in cooperation with institutions located throughout Japan, and have been investigated for their sensitivities to various antibacterial agents and antibiotics and reported by IKEMOTO, et al . Relationships between these isolates and backgrounds of the patients were also studied each year . These results are discussed in detail in this report . In 20 institutions around the entire Japan from October 1991 to September 1992, 631 strains of bacteria were isolated mainly from sputa of 529 patients with respiratory tract infections and tentatively determined to be etiological agents . MICs of various antibacterial agents and antibiotics against 96 strains of Staphylococcus aureus, 112 strains of Streptococcus pneumoniae, 111 strains of Haemophilus influenzae, 114 strains of Pseudomonas aeruginosa (non-mucoid), 41 strains of Moraxella subgenus Branhamella catarrhalis, 39 strains of Pseudomonas aeruginosa (mucoid), Klebsiella pneumoniae and some others, were determined, and the drug sensitivities of these strains were determined except for the strains that had been killed during transportation: 1 . S . aureus . S . aureus strains for which MICs of methicillin were higher than 4 micrograms/ml (methicillin-resistant S . aureus) accounted for 58.3% and the frequency of the drug resistant bacteria increased over previous year's 42.5% . As shown by the MICs, arbekacin was active as vancomycin against all the strains on S . aureus . 2 . S . pneumoniae: Benzylpenicillin among the penicillins showed a potent activity against S . pneumoniae . Cefuzonam, cefmenoxime, cefozopran and cefotaxime among the cephems showed excellent antimicrobial activities against S . pneumoniae . Imipenem; a penem antibiotic, showed the most potent activity with MIC80 of 0.03 micrograms/ml . 3 . H . influenzae: Activities of all drugs were excellent against H . influenzae strains tested . Ampicillin showed MIC80 of 1 micrograms/ml against H . influenzae . Cefuzonam showed the most potent activity among cephems, it completely killed all bacteria at MIC 0.06 micrograms/ml . Cefotaxime and cefmenoxime showed next most potent activities with MIC80s of 0.06 micrograms/ml . The antimicrobial activity of ofloxacin was equivalent to those of cephems . 4 . P . aeruginosa (mucoid) . Ciprofloxacin and tobramycin showed the most potent activities against P . aeruginosa (mucoid), and their MIC80s were 4 micrograms/ml . 5 . P . aeruginosa (non-mucoid): Similarly, ciprofloxacin and tobramycin showed the most potent activities against P . aeruginosa (non-mucoid) with MIC80 of 2 micrograms/ml . Comparing to activities against P . aeruginosa (mucoid), all the drugs tested showed lower activities against P . aeruginosa (non-mucoid) . 6 . K . pneumoniae: The activities of all drugs except for penicillins were very high against K . pneumoniae.(ABSTRACT TRUNCATED AT 400 WORDS)

Jpn J Antibiot, 1995 Aug, 48(8), 1026 - 32
{Study of clinically isolated new quinolones-resistant Haemophilus influenzae . Part 1}; Suzuki Y et al.; A study was done to determine susceptibilities of Haemophilus influenzae that were obtained in our laboratory in 1994 to new quinolones (NQ) and other drugs . The results were as follows; 1 . Among the 300 isolates, the detection frequency of NQ-resistant strains was 8.7% (26 strains), including isolates from chronic lower respiratory tract infections (22 strains) and those from middle meatus of nose (2 strains), etc . NQ-resistant strains were not isolated from children . 2 . The cross resistance was studied for different NQs against NQ-resistant strains . Clavulanic acid/amoxicillin, cefteram, cefpodoxime, cefditoren, cefodizime (CDZM) and cefpirome showed strong antimicrobial activities against NQ-resistant strains . MIC90 of CEPs against all isolated strains including NQ-resistant strains and beta-lactamase producers was low . And the MIC90 of CDZM was < or = 0.025 microgram/ml, which was the lowest among all the antibiotics tested . 3 . We found 47 strains (15.7%) of beta-lactamase producers among the 300 isolates, the frequency of beta-lactamase producing strains was high among strains obtained from children.

MMWR Morb Mortal Wkly Rep, 1995 Jul 28, 44(29), 545 - 50
Progress toward elimination of Haemophilus influenzae type b disease among infants and children--United States, 1993-1994; Frequency and distribution of DNA uptake signal sequences in the Haemophilus influenzae Rd genome; Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USAThe naturally transformable, Gram-negative bacterium Haemophilus influenzae Rd preferentially takes up DNA of its own species by recognizing a 9-base pair sequence, 5'-AAGTGCGGT, carried in multiple copies in its chromosome . With the availability of the complete genome sequence, 1465 copies of the 9-base pair uptake site have been identified . Alignment of these sites unexpectedly reveals an extended consensus region of 29 base pairs containing the core 9-base pair region and two downstream 6-base pair A/T-rich regions, each spaced about one helix turn apart . Seventeen percent of the sites are in inverted repeat pairs, many of which are located downstream to gene termini and are capable of forming stem-loop structures in messenger RNA that might function as signals for transcription termination.

Science, 1995 Jul 28, 269(5223), 496 - 512
Whole-genome random sequencing and assembly of Haemophilus influenzae Rd; Fleischmann RD et al.; An approach for genome analysis based on sequencing and assembly of unselected pieces of DNA from the whole chromosome has been applied to obtain the complete nucleotide sequence (1,830,137 base pairs) of the genome from the bacterium Haemophilus influenzae Rd . This approach eliminates the need for initial mapping efforts and is therefore applicable to the vast array of microbial species for which genome maps are unavailable . The H . influenzae Rd genome sequence (Genome Sequence DataBase accession number L42023) represents the only complete genome sequence from a free-living organism.

Dtsch Med Wochenschr, 1995 Jul 7, 120(27), 953 - 8
{Nodular pulmonary opacities caused by a severe suppurative bronchitis and bronchiectases}; Ewig S et al.; HISTORY AND CLINICAL FINDINGS: A 67-year-old man with chronic obstructive pulmonary disease (COPD) for many years and a heavy smoker was hospitalized because of increasing dyspnoea and moderately productive cough . His general condition was clearly impaired . He was markedly cyanotic, he had a barrel-shaped chest, and there were wheezing rhonchi throughout the lung . Body temperature was 37.8 degrees C . He had mild ankle oedema . The preliminary diagnosis was exacerbation of the COPD with global respiratory failure . TESTS: There was severe hypoxaemia (pO2 48 mm Hg) with hypercapnia (pCO2 46 mm Hg) . Vital capacity was reduced to 1.81, one-second forced expiratory volume 0.91 . Chest radiograph revealed multiple nodular opacities, such as seen in bronchial carcinoma with metastases . But computed tomography showed fluid-filled bronchi . Bronchoscopy demonstrated large amounts of dirty-white, sticky secretions as high up as the trachea, confirming the preliminary diagnosis . Klebsiella oxytoca and haemophilus influenzae were cultured from the secretions . TREATMENT AND COURSE: Immediate administration of amoxycillin and clavulanic acid (2.2 g daily) quickly led to clinical improvement . Chest x-ray was normalized after 10 days . But long-term oxygen treatment was instituted because of persisting resting hypoxaemia (pO2 46 mm Hg), after which he was discharged free of dyspnoea . CONCLUSION: To clarify nodular pulmonary opacities in a chest radiograph computed tomography should be performed before bronchoscopy.

Gene, 1995 Jul 4, 160(1), 81 - 6
Construction of Actinobacillus pleuropneumoniae-Escherichia coli shuttle vectors: expression of antibiotic-resistance genes; West SE et al.; We constructed several cloning vectors, designated pGZRS-18/19 and pGZRS-38/39, which were based on an endogenous Actinobacillus pleuropneumoniae (Apl) 4.3-kb plasmid . They carry the lacZ alpha-complementation fragment and MCS from pUC18/19, and either the bla gene under the control of a putative Apl promoter or the KmR gene from Tn903 . These vectors replicate in representative strains of Apl serotypes 1 and 7, Escherichia coli, Pasteurella haemolytica (Ph) and Haemophilus (Actinobacillus) actinomycetemcomitans . We also found that Apl and Ph did not express genes under the control of the lacZ or bla promoters, suggesting that their RNA polymerases may not utilize these promoters.

Microb Drug Resist, 1995 Summer, 1(2), 149 - 58
Pneumonia: the impact of antibiotic resistance on its management; Finch RG; Pneumonia in the community affects between 1 and 5 per 1000 per year . The microbial aetiology is diverse and influenced by preexisting disease, seasonality, as well as animate and inanimate environmental sources; pneumococci, Legionella spp., Mycoplasma pneumoniae, and more recently Chlamydia pneumoniae are the predominant bacterial pathogens . Gram-negative enteric bacteria although less common are particularly virulent . Antibiotic resistance is well established for Haemophilus influenzae and Gram-negative bacillary infections, but has been a recent phenomenon in the case of Streptococcus pneumoniae, which is numerically the leading pathogen . Despite the concerns raised by this reduced susceptibility to penicillin, evidence that this has been translated into increased clinical failures is currently difficult to establish . Macrolide and tetracycline resistance among pneumococci is more common . beta-Lactamase production by H . influenzae has now reached levels where, in those with severe pneumonia, beta-lactamase stable agents are preferred . Consensus Guidelines on the treatment of community acquired pneumonia have been published by the British Thoracic Society, the American Thoracic Society, and from Expert Panels in Canada and France . These emphasize severity assessment and differentiate management in the community or hospital setting . The recommended regimens are compared and contrasted . In conclusion, mild/moderate pneumonia, when pneumococcal in nature, is likely to still respond to amoxycillin or penicillin G, but in higher dosages where pneumococcal resistance is documented . However, in severe infection where pneumococcal resistance, other beta-lactamase-producing pathogens, or an atypical infection could be operating, it is important that initial empirical therapy be broad spectrum and promptly administered . Treating multiresistant pneumococcal disease in those allergic to beta-lactams presents a particular dilemma . Glycopeptides are currently preferred.

Microb Drug Resist, 1995 Summer, 1(2), 131 - 5
Epidemiology of antibiotic resistance in Haemophilus influenzae; Goldstein FW et al.; During the last 10 years, a continuous increase in the incidence of beta-lactamase-producing Haemophilus influenzae has been observed; in addition, beta-lactamase-negative ampicillin-resistant strains are better identified and more frequently isolated . During the same period, resistance to tetracyclines and chloramphenicol decreased to a very low level . The incidence of resistant bacteria is highly variable according to the site of infection, patient's age, type of Haemophilus, and country or region, according to local epidemiological factors . Follow-up multicenter studies are needed to monitor the evolution of resistance to these antibiotics and also emergence and spread of resistance to other antibiotics, such as new fluoroquinolones, new beta-lactams, and new macrolides.

Rev Latinoam Microbiol, 1995 Jul-Sep, 37(3), 189 - 99
{Haemophilus influenzae infections in 2 hospitals in the city of Puebla, Mexico}; Sosa-Iglesias EG et al.; Haemophilus influenzae has been recognized as one of the most important pathogen in the pediatric population younger than 5 years old . We analized some characteristics of the infectious diseases by Haemophilus influenzae in the pediatric group from 1985 to 1990 in two hospitals of Puebla city . From 321 children studies cases included for this work, fifty of those had infectious diseases by H . influenzae . Forty six percent of the patients developed meningitis, 22% otitis media, 12% sepsis, 10% pneumonia and 10% other infectious diseases . The majority of cases occurred in males but the difference was not statistically significant (p > 0.05) . Eighty percent of diseases by H . influenzae occurred in infants younger than 24 months . The serovar b was the most frequent in invasive infections and nontypable strains were frequent in otitis media . The biovars I and IV were isolated from invasive infections . During this study two children died with meningitis and 3 patients had severe neurological damage . This finding suggests that is necessary to vaccine this population to prevent the morbilethality of infectious diseases by Haemophilus influenzae.

Ann Ig, 1995 Jul-Aug, 7(4), 235 - 41
{Epidemiology of invasive infections with Haemophilus influenzae type b in the world and in Italy}; Tozzi AE et al.; The incidence of Haemophilus influenzae type b (Hib) disease in the 0-5 years age group has been studied in many countries . In the United States, before the introduction of mass vaccination, the incidence of invasive Hib disease ranged from 7 per 100,000 in the 4-5 years age group to 452 pert 100,000 in the 6-11 months age group . In Europe, the incidence of this disease has been estimated to range from 21 to 60 per 100,000 per year in the 0-5 years age group . Hib infection is mainly transmitted by the respiratory route . Risk factors of the disease include: attending a day-care center, a high number of household members, low socio-economic level, age less than two years, and belonging to certain ethnic groups . Accurate estimates of the incidence of invasive Hib disease do not exist in Italy . Among the 15,601 children participating in the "Progetto Pertosse", a clinical trial for the evaluation of antipertussis vaccines, there occurred six cases of Hib meningitis, one of Hib sepsis, and one of Hib cellulitis . These episodes yield an incidence density of 28.7 per 100,000 person-years in the 2-30 month age group . Incidence data will also need to be collected for children in the 30 months-5 years age group before attempting a cost-benefit analysis with the aim of planning a mass vaccination.

Antibiot Khimioter, 1995 Jul, 40(7), 35 - 8
{Dirithromycin in the treatment of infections of the lower respiratory tract}; Belousov IuB et al.; Good results of the treatment of patients with lower respiratory tract infections with dirithromycin (Eli Lilly, USA), a new semisynthetic macrolide, were recorded . The trial included 15 patients: 6 with acute bronchitis (AB) and 9 with exacerbation of chronic bronchitis (AB) and 9 with exacerbation of chronic bronchitis (ECB) . The antibiotic was administered orally in a single dose of 500 mg once a day for 7 days . The treatment efficacy was estimated by the clinical results and laboratory findings . The sputum specimens were investigated bacteriologically with testing the microflora for the drug susceptibility by using the diffusion disks . 50 per cent of the patients with AB isolated Streptococcus viridans with low (the diameter of the growth inhibition zones < 16 mm) and intermediate (16-17 mm) susceptibility to the antibiotic, 33.3 per cent of the patients isolated highly susceptible (19 mm) strains of Str . pneumonia and 16.7 per cent of the patients isolated resistant (15 mm) strains of Staphylococcus epidermidis . Highly susceptible (17-19 mm) strains of Haemophilus influenzae and Str . viridans were isolated respectively from 55.5 and 45.5 per cent of the patients with ECB . In 4 patients with ECB a clinical improvement of the state was recorded . In the other patients with AB and ECB the recovery was stated . The bacteriological tests revealed a new pathogen in 2 patients with ECB and the failure of the treatment in another 2 patients with ECB . In all the other patients the pathogen was shown to be eradicated . In the patients isolating the new pathogen the symptomatic recovery was stated in the posttherapeutic period . Therefore, diritromycin proved to be efficient in 13 out of the 15 patients with lower respiratory tract infections . It should be noted that the drug tolerance was excellent . None of the patients showed any adverse reactions.

Diagn Microbiol Infect Dis, 1995 Jul, 22(3), 301 - 6
Comparative in vitro activity of furopenem against aerobic bacteria isolated from pediatric patients; Mortensen JE et al.; The in vitro activity of the oral penem furopenem (WY-49605, 545555, SUN5555, and ALP201) was tested against clinical bacteria isolated from pediatric patients . Furopenem was compared with clarithromycin, cefpodoxime, amoxicillin, amoxicillin-clavulanate, cefaclor, cefixime, and cefuroxime . Furopenem demonstrated consistent activity against Escherichia coli {minimum inhibitory concentration (MIC90) = 1.0 microgram/ml)} Klebsiella pneumoniae (MIC90 = 2.0 micrograms/ml), Salmonella enteriditis and Shigella spp . (MIC90 = 1.0 microgram/ml), and beta-lactamase-positive or -negative Haemophilus influenzae (MIC90 = 1.0 microgram/ml) and Moraella catarrhalis (MIC90 = 1.0 microgram/ml) . Furopenem was also active against a number of the Gram-positive organisms tested including methicillin-susceptible Staphylococcus aureus and penicillin-susceptible Streptococcus pneumoniae . These results suggest a potential application for this agent in the treatment of children as outpatients.

Avian Dis, 1995 Jul-Sep, 39(3), 507 - 13
The effect of route of inoculation on protection by killed vaccines in chickens; Nakamura T et al.; The effect of various routes of immunization on protection against challenge by virulent agents was examined in chickens . Chickens were immunized intratracheally, intranasally, per os, by crop gavage, and intramuscularly . Agents examined were killed Haemophilus paragallinarum, Mycoplasma gallisepticum, and infectious bursal disease virus . Results of immunization by intratracheal administration were equivalent to those produced by parenteral administration . All vaccines effectively induced production of serum antibodies against pathogens, and all immunized chickens were protected against infection by each pathogen.

Microb Pathog, 1995 Jul, 19(1), 31 - 8
Specificity of the immune response to Haemophilus ducreyi; Brown TJ et al.; The specificity of the antibody response to Haemophilus ducreyi in sera from patients attending a sexually transmitted disease clinic in South Africa has been studied using immunoblotting . Patients with chancroid were shown to have higher levels of IgG (mean 0.74, SD 0.34) to H . ducreyi than those with no history of chancroid (mean 0.34, SD 0.19) . The pattern of the antibody specificity was highly variable between patients with culture proven chancroid but there was no observed strain specificity . In comparison, the patterns obtained using sera from patients without known exposure to H . ducreyi showed less variation between patients and were of less intensity at the dilution used . Sera from patients with chancroid recognised epitopes on proteins that varied in molecular weight between strains, particularly of 60-66kDa (10 of 36 patients) and 25-27kDa (8 of 36 patients) . In addition epitopes were recognised on the GroEL and/or DnaK heat shock proteins in 13 of 36 sera tested . There was no apparent change in the epitopes recognised on proteins between the homologous and heterologous strains . Patterns of antibody specificity in sequential sera only varied in one of six patients tested.

J Antimicrob Chemother, 1995 Jul, 36 Suppl A, 73 - 84
Efficacy of meropenem in experimental meningitis; Nairn K et al.; Meropenem and comparator antibiotics, including ceftriaxone, ceftazidime, benzyl penicillin and a combination of ampicillin plus gentamicin, were evaluated in a model of bacterial meningitis in the guinea-pig . The model is an acute infection in which challenge with each organism, if untreated, causes an increase in numbers of white blood cells, elevation of protein concentrations and 6-8 log10 cfu/mL of bacteria in the CSF . Infections caused by Haemophilus influenzae, Neisseria meningitidis, three strains of Streptococcus pneumoniae (two penicillin-resistant), Escherichia coli, Pseudomonas aeruginosa and Listeria monocytogenes all responded to meropenem, which was as active as the comparator agents in all studies, and was more active in most . Of particular note were the results seen against S . pneumoniae (penicillin-resistant) infections, in which meropenem was significantly more effective than ceftriaxone . Also notable were results from the P . aeruginosa infection where meropenem, at low doses, was more effective than ceftazidime . Activity against L . monocytogenes was equivalent to that produced by treatment with the combination of ampicillin plus gentamicin, even when treatment was delayed . These results show that, in an animal model, meropenem penetrates into CSF in concentrations sufficient to produce significant reductions in the numbers of common and less common pathogens.

J Antimicrob Chemother, 1995 Jul, 36(1), 173 - 83
A comparison of 5 days of dirithromycin and 7 days of clarithromycin in acute bacterial exacerbation of chronic bronchitis; Hosie J et al.; The safety and efficacy of dirithromycin and clarithromycin were compared in a single-blind, multicentre study of patients with acute bacterial exacerbation of chronic bronchitis (AECB) . Patients received either dirithromycin, 500 mg once daily for 5 days, or clarithromycin, 250 mg twice daily for 7 days . A total of 212 patients entered the study, of whom 191 qualified for efficacy analysis . Favourable post-therapy clinical and bacteriological response rates for qualified patients (95 dirithromycin and 96 clarithromycin) were similar: 89.5% and 68.8% for dirithromycin vs 94.8% and 71.9% for clarithromycin . At late post-therapy evaluation, favourable clinical and bacteriological response rates were achieved in 98.8% and 96.2% of dirithromycin patients and 95.3% and 93.3% of clarithromycin patients, respectively . These differences were neither statistically nor clinically significantly different . Both drugs had similar efficacy against Haemophilus influenzae and both were well tolerated . Dirithromycin, administered as a single daily dose for just 5 days resulted in complete compliance in all but four patients . In clarithromycin-treated patients, requiring a 7-day course of twice-daily treatment, compliance was less satisfactory, with 12 patients failing to comply, though the between-group difference was not statistically significant . It can be concluded that 5 days of dirithromycin, 500 mg once daily is as safe and effective as 7 days of clarithromycin, 250 mg, twice daily in the treatment of AECB.

Biopharm Drug Dispos, 1995 Jul, 16(5), 393 - 401
Evaluation of the effects of ambroxol on the ofloxacin concentrations in bronchial tissues in COPD patients with infectious exacerbation; Paganin F et al.; Infectious excerbations of COPD are generally due to Streptococcus pneumoniae, Haemophilus species, and other Gram-negative bacteria . Ofloxacin has potent activity against Gram-negative species but is less effective against Gram-positive species including Streptococcus pneumoniae . It has also been shown that the administration of ambroxol increases the concentration of some antibiotics in pulmonary tissues . The aim of the study was to determine whether ambroxol increases the bronchial tissue concentrations of ofloxacin to a level exceeding the MIC90 of the bacterial species less susceptible to ofloxacin . 24 patients with COPD were randomized in two groups . Drug regimens of ofloxacin alone (200 mg twice daily) or ofloxacin (200 mg twice daily)+ambroxol (30 mg thrice daily) were administered over 10 d . A fibroscopy was performed on day 10 with bronchial biopsies and broncho-alveolar lavage . At steady state, concentrations of drug in plasma and bronchial samples were assayed by HPLC with fluorometric detection . There was no significant difference in the bronchial levels of ofloxacin between the two groups; however, in alveolar cells, ofloxacin concentration was three times higher in the group with ambroxol . Ambroxol does not increase ofloxacin concentrations in bronchial tissue because high concentrations are already present in the lung.

Can J Vet Res, 1995 Jul, 59(3), 173 - 8
A comparative study of bovine and ovine Haemophilus somnus isolates; Ward AC et al.; Bacterial isolates (including 17 Haemophilus somnus isolates and an H . somnus-like isolate) from asymptomatic or diseased cattle and sheep, were evaluated for markers associated with virulence and host predilection . The isolates were separated into 6 distinct biovariants, 3 for sheep and 3 for cattle, based on reactions in a battery of 21 test media . Three bovine isolates associated with disease caused hemolysis of bovine blood . The rest of the isolates did not hemolyze either bovine or ovine erythrocytes . Protein profiles of all H . somnus isolates were similar with the exception of the major outer membrane proteins (MOMPs) . The MOMPs of isolates associated with disease in cattle had a relative molecular weight of approximately 41 kDa compared with 33 kDa for the MOMPs of isolates from asymptomatic cattle . The MOMPs from sheep isolates were either slightly higher or lower than the 41 kDa MOMPs of bovine isolates . Major antigens detected by Western blotting were similar in all isolates except the H . somnus-like isolate . An immunodominant 40 kDa antigen was conserved in all H . somnus isolates . Antibodies to this antigen have previously been found to be protective in cattle and may also be protective for sheep . Marked differences between cattle and sheep isolates were revealed by use of restriction enzyme analysis, which separated the isolates into 12 ribotypes and 15 unique DNA profiles . Thus, cattle and sheep isolates in this collection had distinctive differences in biochemical reactions, MOMP profiles, and DNA analyses . Such differences have potential value for epidemiological studies and may also be used to evaluate host specificity of H . somnus isolates.

J Infect Dis, 1995 Jul, 172(1), 132 - 5
Epidemiology of nasopharyngeal colonization with nontypeable Haemophilus influenzae in the first 2 years of life; Faden H et al.; Two hundred children were followed from birth through 2 years of age with nasopharyngeal cultures to determine the normal colonization pattern of nontypeable Haemophilus influenzae . Forty-four percent of the children were colonized on one or more occasions; the acquisition rate was greatest in the first year . Monthly prevalence rates were 11% . Colonization with the initial strain persisted 1-5 months (median, 2) . Children carried 1 predominant strain at a time but became colonized with up to 7 different strains (mean, 2.2) . Children colonized with a single strain for < or = 2 months produced a greater nasopharyngeal secretory IgA to nontypeable H . influenzae response than did children colonized with different strains (log 2.35 +/- 0.68 vs . 1.89 +/- 0.25 U of P6 secretory IgA/ng/mL of total secretory IgA, P < .01) . The duration of colonization with a strain and acquisition of a new strain may be affected in part by the local production of specific secretory IgA.

Infect Immun, 1995 Jul, 63(7), 2485 - 92
Binding of Haemophilus influenzae to purified mucins from the human respiratory tract; Davies J et al.; Mucins are high-molecular-weight glycoproteins and major constituents of the mucus layer which covers the airway surface . We have studied the interactions between bacteria, mucins, and epithelial cells from the human respiratory tract . Nontypeable strains of Haemophilus influenzae were found to bind to purified airway mucins in suspension and on solid phase . Mucins in suspension inhibited the attachment of these strains to nasopharyngeal epithelial cells, while mucin coating of the cells enhanced their binding . In contrast, strains of Streptococcus pneumoniae and encapsulated and other nontypeable H . influenzae strains failed to interact with mucins . These H . influenzae strains used other strategies for adherence to epithelial cells . The type b strain 770235 attached via fimbriae but also expressed a subcapsular adhesin that was detected in a capsule- and fimbria-defective mutant . Mucin pretreatment of these bacteria did not inhibit adherence, but mucin pretreatment of epithelial cells inhibited adherence, probably by shielding of the receptors for these adhesins . Non-mucin-binding nontypeable and encapsulated H . influenzae strains would, therefore, adhere only after disruption of the mucus layer and exposure of cellular receptors . Differences in tissue toxicity and invasiveness among H . influenzae strains may also be influenced by the mucin interactions of the strains.

Infect Immun, 1995 Jul, 63(7), 2467 - 72
Cellular immunity to the P6 outer membrane protein of nontypeable Haemophilus influenzae; Kodama H et al.; Cellular immunity to nontypeable Haemophilus influenzae in a population of 10 healthy, immune adults was determined by measuring lymphocyte blast transformation and antibody secretion in response to the P6 outer membrane protein . P6 (200 microliters/ml) induced lymphocyte blast transformation that peaked on day 10 of incubation . The peak induction of antibody-secreting cells occurred on day 8 of incubation . In comparison with the response to tetanus toxoid stimulation, the peak lymphocyte blast transformation response to P6 was reduced (mean counts per minute +/- standard error of the mean {SEM}, 3,457 +/- 503 versus 9,414 +/- 1,464; P = 0.0051) and delayed (mean days +/- SEM, 10.3 +/- 0.4 versus 8.4 +/- 0.5; P = 0.0169); however, P6 was a better stimulus of antibody secretion from lymphocytes, particularly antibody of the immunoglobulin M (IgM) class (mean peak numbers of antibody-secreting cells per 10(5) peripheral blood mononuclear cells +/- SEM: IgG, 85 +/- 29 versus 42 +/- 16 {P = 0.0469}; IgM, 81 +/- 20 versus 25 +/- 7 {P = 0.0125}; IgA, 24 +/- 8 versus 16 +/- 6 {P = 0.0526}) . Thus, lymphocytes from immune individuals recognize P6 of nontypeable H . influenzae as an immunogen . These data provide a basis for future studies with otitis-prone children who fail to develop a normal antibody response to P6 antigen (N . Yamanaka and H . Faden, J . Pediatr . 122:212-218, 1993).

J Fla Med Assoc, 1995 Jul, 82(7), 467 - 9
Changing epidemiology of childhood meningitis; Subedar N et al.; The changing epidemiology of meningitis at Wolfson Children's Hospital in Jacksonville between 1985 and 1994 was reviewed to determine the impact of Haemophilus influenzae type b vaccination . The cases were also straified by age . A significant decrease in the cases of Haemophilus influenzae type b meningitis was found between 1991 and 1994 . This was most likely due to the introduction of conjugate Haemophilus influenzae type b vaccine for young infants.

J Clin Microbiol, 1995 Jul, 33(7), 1835 - 8
High incidence of Haemophilus influenzae in nasopharyngeal secretions and middle ear effusions as detected by PCR; Ueyama T et al.; PCR was used to detect Haemophilus influenzae in samples of nasopharyngeal secretion and middle ear effusion (MEE) . Nasopharyngeal secretions were collected from 102 patients with otitis media with effusion and from 111 healthy subjects . Eighty samples of MEE were collected from patients with otitis media with effusion . A pair of primers was designed to amplify a DNA segment of the gene encoding P6 outer membrane protein of H . influenzae . The amplified PCR product was detected with an internal probe that hybridized specifically to the P6 DNA of H . influenzae . Samples of MEE and nasopharyngeal secretion were also examined by a conventional culture method . The incidence of P6 gene DNA in nasopharyngeal secretions detected by PCR was about two times higher than that of H . influenzae detected by the conventional culture . Culture-positive samples were all positive in the PCR test . In MEEs, the rate of detection of the P6 gene DNA target was about five times higher than that of H . influenzae detected by the culture method . All patients who had P6 gene DNA in MEEs were found to have the DNA in nasopharyngeal secretions . These findings suggest that the presence of H . influenzae in MEEs and in nasopharyngeal secretions is more common than previously reported.

J Med Microbiol, 1995 Jul, 43(1), 45 - 9
Clonal analysis of Haemophilus influenzae type b isolates in the United Kingdom; Urwin G et al.; Strains of Haemophilus influenzae type b were collected as part of an epidemiological study of bacterial meningitis in the North East Thames Region (NETR) of England . Subclones of H . influenzae were identified by outer-membrane protein typing and multilocus enzyme electrophoresis . These were compared with subclones of H . influenzae type b isolated from elsewhere in the UK . The subclone ET 12.5/OMP 3L was identified in 68% of isolates from cases of meningitis from the NETR . In the isolates from elsewhere in the UK, this subclone was identified in 79% of sterile-site isolates and 91% of isolates from non-sterile sites . This subclone predominated in patients of different ages and from different ethnic groups, which suggests that non-Caucasians are infected with the subclone predominating locally . Since ET 12.5/OMP 3L subclone is the most abundant strain isolated in virtually all other Western European countries, these results provide additional evidence that the genetic diversity in the H . influenzae type b population in Europe is relatively restricted.

J Pediatr, 1995 Jul, 127(1), 68 - 75
Immunodeficiency in long-term survivors of acute lymphoblastic leukemia treated with Berlin-Frankfurt-Münster therapy; Smith S et al.; OBJECTIVE: To determine whether the immune systems of long-term survivors of acute lymphoblastic leukemia (ALL) have persistent immune defects after Berlin-Frankfurt-Munster (BFM) treatment . STUDY DESIGN: We evaluated the cellular and humoral immune responses of 13 children with ALL in complete remission and off modified protocol treatment for 2 or more years . All patients had received complete immunizations for measles, mumps, rubella, and poliovirus before ALL developed . They were challenged with Haemophilus influenzae type B (Hib) and Pneumococcus vaccines after baseline serum samples were obtained . We also determined in vivo humoral immune responses to bacteria and viruses that cause common pediatric diseases . RESULTS: Compared with age-matched control subjects, the long-term survivors of ALL had a significant difference in the presence of protective antibodies to measles (p < 0.0001) and polioviruses (p < 0.0001) in their baseline sera; more than half had no protective antibodies to one or more previously administered vaccines or related infections . Most produced protective concentrations of specific antibody after reimmunization, but some were repeatedly unable to make protective antibodies, or mount a normal antibody response, despite natural disease and/or revaccination . Four children had significant infections . CONCLUSIONS: Long-term survivors of ALL who had BFM treatment may have persistent immune defects with respect to common childhood bacterial and viral diseases they previously had, or vaccines they received.

Am J Respir Crit Care Med, 1995 Jul, 152(1), 169 - 73
Progression of pulmonary disease after disappearance of Pseudomonas in cystic fibrosis; Sharma GD et al.; Once cystic fibrosis (CF) patients become chronically colonized, eradication of Pseudomonas aeruginosa (PA) is rare . We report five patients, each colonized for at least 6 yr, whose subsequent cultures did not reveal PA or any other gram-negative pathogen for at least 2 yr . Two patients harbored yeast, normal throat flora, and occasional colonies of Aspergillus fumigatus, but no PA, Haemophilus influenzae, or other gram-negative pathogens . In two patients, sputum cultures revealed Staphylococcus aureus . Sputum smear showed no gram-negative organisms in any patient . Antipseudomonal antibody titers in all patients decreased during the noncolonized period . However, titers increased in four other CF patients who had continued PA colonization . Despite disappearance of PA, four of five patients had clinical progression of lung disease with deterioration of FEV1 . We conclude that (1) PA, even after it has chronically colonized the airways of CF patients, can occasionally disappear; and (2) lung disease can progress despite the disappearance of PA . Thus, the elimination of a milieu favorable to PA, such as might be anticipated with therapy directed at the basic defect, may not be sufficient to halt the pulmonary disease progression in CF.

Pediatrics, 1995 Jul, 96(1 Pt 1), 18 - 22
Immunologic response of extremely premature infants to tetanus, Haemophilus influenzae, and polio immunizations; D'Angio CT et al.; OBJECTIVE . To determine whether extremely premature infants have immunologic responses to tetanus toxoid, Haemophilus influenzae type b polysaccharide and polio vaccines similar to those of full-term infants . INFANTS AND METHODS . Sixteen extremely premature (< 29 weeks, < 1000 g at birth) infants received separate diphtheria-tetanus-pertussis and H influenzae type b oligosaccharide-CRM197-conjugated (HbOC) vaccines at 2, 4 and 6 months of chronologic age, enhanced potency inactivated polio vaccine at 2 months, and oral polio vaccine at 4 months . Serum was obtained for anti-tetanus toxoid (TT), anti-Haemophilus b polysaccharide (HbPs) and polio neutralizing antibody assays before the 2-month vaccination and 4 to 6 weeks after the 6-month vaccination . Comparison sera were obtained from full-term infants immunized with the same lots of diphtheria-tetanus-pertussis (n = 46) and HbOC (n = 66) vaccines or the same sequence of polio vaccines (n = 10) . RESULTS . Preterm and full-term infants had similar geometric mean titers of anti-TT antibodies, anti-HbPs antibodies, and neutralizing antibodies to polio serotypes 1, 2, and 3 after the completion of the primary series of vaccines . After vaccination, similar proportions of preterm and full-term infants had protective levels of antibody to TT (preterm 100% vs full-term 100% with levels > 0.01 IU/mL), HbPS (82% vs 87%, > 1.0 microgram/mL), and polio serotypes 1 (85% vs 80%, > or = 1:8) and 2 (100% vs 100%, > or = 1:8) . Preterm infants were less likely than full-term infants to have protective levels of neutralizing antibody to polio serotype 3 (31% vs 90%, > or = 1:8) . CONCLUSIONS . Extremely premature infants have adequate antibody responses to tetanus and HbOC antigens but may have diminished responsiveness to serotype 3 polio vaccine.

Eur J Clin Microbiol Infect Dis, 1995 Jul, 14(7), 648 - 54
Outpatient treatment of endocarditis in a clinic-based program in Argentina; Stamboulian D; The major cost in the treatment of infective endocarditis (IE) is the length of hospitalization required for the administration of intravenous antibiotics . This can be reduced by substituting shorter regimens and by the introduction of outpatient parenteral antibiotic therapy (OPAT) . Careful selection of patients is vital for the success of OPAT in IE . The patients should be hemodynamically stable and without clinical complications . The delivery of OPAT for IE followed a clinical and home-based program involving an endocarditis team whose members included an infectious diseases physician, a microbiologist, a cardiologist and a nurse trained in intravenous techniques . Among the antimicrobial agents used in OPAT of IE, single-agent ceftriaxone for four weeks followed by a short course of amoxicillin or ceftriaxone in combination with an aminoglycoside for two weeks (short course) are effective modes of treatment for streptococcal endocarditis, the most common cause of IE . This treatment is also effective for carefully selected patients with other types of endocarditis, such as those due to the HACEK group (Haemophilus aphrophilus/paraphrophilis, Actinobacillus actinomycetem comitans, Caradiobacterium hominis, Eikenella corrodens and Kingella kingae) . Staphylococcus aureus, enterococci and late prosthetic valve endocarditis associated with a streptococcus may also be treated on an outpatient basis after stabilization (approximately 2 weeks) . As a result of their need for prolonged treatment periods, these patients are also very good candidates for OPAT . In conclusion, new regimens utilizing ceftriaxone once daily and short-term therapy on a clinical or home basis offer the potential benefits of cheaper safer and more convenient treatment for patients with IE.

Can Vet J, 1995 Jul, 36(7), 424 - 9
Effects of various vaccination protocols on passive and active immunity to Pasteurella haemolytica and Haemophilus somnus in beef calves; Van Donkersgoed J et al.; Two field trials were conducted in a beef cow herd in Saskatchewan to determine the effectiveness of a combined Pasteurella haemolytica and Haemophilus somnus vaccine in increasing passively and actively acquired antibodies in beef calves . Vaccination of dams at 4 and/or 7 weeks prepartum was associated with increased antibody titers to P . haemolytica and H . somnus in their serum (P < 0.05), colostrum (P < 0.05), and serum of their calves at 3 days and 1 month of age (P < 0.05) . There was no significant (P > 0.05) difference in antibody titers in the colostrum and serum of calves from single or double vaccinated dams . Calves vaccinated at 1 and 2 months of age in the face of maternal antibodies to P . haemolytica and H . somnus had significantly (P < 0.05) higher antibodies to P . haemolytica and H . somnus at 4 and 6 months of age than did unvaccinated calves . Calves vaccinated at 3 and 4 months of age in the face of low levels of preexisting antibodies had significantly (P < 0.05) higher antibodies to P . haemolytica at 5 months of age and to H . somnus at 5 and 6 months of age than did unvaccinated calves . Calves vaccinated once at 4 months of age had significantly (P < 0.05) higher antibody titers to P . haemolytica and H . somnus at 4.5 months of age than did unvaccinated calves, but this difference was not apparent at 6 months of age.(ABSTRACT TRUNCATED AT 250 WORDS)

Clin Infect Dis, 1995 Jul, 21(1), 213 - 6
Bacteremia due to Haemophilus infections: a retrospective study with emphasis on the elderly; Najm WI et al.; We performed a retrospective study of all patients in a large health maintenance organization in Southern California who were identified as having positive blood cultures for Haemophilus organisms during a 20-month period (September 1990 to May 1992) to assess the incidence, presentation, and predisposing conditions of bacteremia due to these organisms and to examine some of the features of these infections in the elderly . Thirty-eight patients with bacteremia due to haemophilus infections were identified . Ten (26.3%) patients were 65 years of age or older . The incidence of bacteremic haemophilus infections in the elderly group was estimated at 2.7 per 100,000 individuals per year, which was almost three times greater than that for the younger age groups studied . When analyzed statistically, the presenting feature of the infection did not differ among age groups . Six patients died, four of whom were elderly . All six deaths were due to nontypable Haemophilus influenzae strains . Cancer was the only chronic underlying condition frequently found among the elderly patients . Three of 10 elderly patients lived in nursing homes; all three were infected with nontypable H . influenzae strains, and all three died.

Clin Infect Dis, 1995 Jul, 21(1), 171 - 6
Changes in the core tonsillar bacteriology of recurrent tonsillitis: 1977-1993; Brook I et al.; Microbiological studies of the core of tonsils removed from children with recurrent tonsillitis due to group A beta-hemolytic streptococci were conducted during three periods, with 50 patients in each period: 1977-1978 (period 1), 1984-1985 (period 2), and 1992-1993 (period 3) . Mixed flora were present in all tonsils, with 8.1 organisms per tonsil (3.8 aerobes and 4.3 anaerobes) . The predominant isolates in each period were Staphylococcus aureus, Moraxella catarrhalis, Peptostreptococcus species, pigmented Prevotella species, Porphyromonas species, and Fusobacterium species . The rate of recovery of Haemophilus influenzae type b increased from 24% in period 1 to 76% in period 2 (P < .001); a decline to 12% in period 3 correlated with a concomitant increase in the frequency of recovery of non-type b strains of H . influenzae from 4% and 10% in periods 1 and 2, respectively, to 64% in period 3 (P < .001) . Both the rate of recovery of beta-lactamase-producing bacteria and the number of these organisms per tonsil increased over time . Specifically, beta-lactamase-producing strains were detected in 37 tonsils (74%) during period 1, in 46 tonsils (92%) during period 2, and in 47 tonsils (94%) during period 3, and the number of such strains per tonsil increased from 1.1 in period 1 to 2.9 and 3.3 in periods 2 and 3, respectively.

J Vet Diagn Invest, 1995 Jul, 7(3), 313 - 20
Porcine reproductive and respiratory syndrome: NEB-1 PRRSV infection did not potentiate bacterial pathogens; Cooper VL et al.; A 2-phase study was conducted to evaluate the ability of the NEB-1 strain of porcine reproductive and respiratory syndrome virus (PRRSV) to potentiate common bacterial pathogens of swine . In phase I, 25 of 50 4-5-week-old specific-pathogen-free (SPF) pigs were exposed to NEB-1 PRRSV (day 0) . Seven days after virus inoculation, 8 groups received 1 of 4 bacterial pathogens: Haemophilus parasuis, Streptococcus suis, Salmonella cholerasuis, and Pasteurella multocida . The ability of NEB-1 PRRSV to produce clinical disease, viremia, neutralizing antibody, gross and microscopic lesions and to potentiate bacterial pathogens was assessed . Response to NEB-1 PRRSV was similar among inoculated pigs; prolonged hyperthermia, lethargy, mild to moderate dyspnea, and cutaneous erythema were consistent clinical signs . No clinical differences were observed in groups after bacterial challenge . Virus was isolated from serum at weekly intervals through the end of the study, and all PRRSV-inoculated pigs had seroconverted by study termination . Two of 5 pigs died in non-PRRSV-inoculated groups challenged with H . parasuis and Streptococcus suis . Mortality in PRRSV-infected pigs was limited to 1 of 5 pigs from the Salmonella cholerasuis-challenged group . Gross lesions were seen in pigs dying after inoculation in H . parasuis- and Streptococcus suis-inoculated groups, in Salmonella cholerasuis- and P . multocida-challenged pigs, and in 1 non-PRRSV-inoculated control pig . Microscopic lesions consisted of mild to moderate proliferative interstitial pneumonia, nonsuppurative myocarditis, lymphoid hyperplasia, and nonsuppurative encephalitis in PRRSV-inoculated pigs . Findings in phase I indicated that NEB-1 PRRSV does not potentiate bacterial disease while inducing consistent clinical signs, viremia, seroconversion, and microscopic lesions.(ABSTRACT TRUNCATED AT 250 WORDS)

APMIS, 1995 Jul-Aug, 103(7-8), 540 - 7
Development of a peroxidase-antiperoxidase (PAP) technique for the identification of Haemophilus somnus in pneumonic calf lungs in Denmark; Tegtmeier C et al.; A peroxidase-antiperoxidase (PAP) technique was developed for the identification of Haemophilus somnus bacteria in lung tissues of calves . Antisera raised against somatic and wall antigens of a Danish and American strain of H . somnus were produced . Experimentally infected murine tissues were used for the determination of the sensitivity and specificity of antiserum that had been heterologously absorbed with antigens of cross-reacting bacteria, i.e . Pasteurella haemolytica and Pasteurella multocida . None of the antisera reacted with Actinomyces pyogenes . An antiserum raised against somatic antigens of the Danish strain of H . somnus revealed the highest sensitivity in the PAP technique and became specific following absorption . Heterologous absorption also rendered this antiserum specific in crossed immunoelectrophoresis . Subsequently, the PAP technique was applied on formalin-fixed pneumonic lung tissues of 86 calves . An immunodiagnosis of H . somnus pneumonia was obtained in 15 of 17 lungs from which the bacterium had been isolated . Moreover, immunostained bacteria were also demonstrated in 20 lungs from which H . somnus had not been isolated . Thus, application of immunohistochemistry significantly enhanced the diagnostic sensitivity of H . somnus pneumonia of calves and should be used as a potent supplementary tool for the routine screening of suspected lung tissues of calves from which bacterial isolation is negative.

Pediatr Infect Dis J, 1995 Jul, 14(7 Suppl), S84 - 7
Postantibiotic effect of ceftibuten on respiratory pathogens; Chin NX et al.; The postantibiotic effect (PAE) of ceftibuten, a novel beta-lactamase-stable cephem, was determined for Streptococcus pyogenes, Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis . The ceftibuten PAE after a 2-hour exposure to 2 micrograms/ml (4 x minimum inhibitory concentration) for S . pyogenes was 2.7 to > 10 hours . The PAE for S . pneumoniae after a 2-hour exposure to 15 micrograms/ml, concentrations that are achieved in man after usual therapeutic doses, was 1.1 to 3.4 hours and the PAE for H . influenzae was 1 to 1.1 hours . M . catarrhalis had a PAE of 1.5 to 1.8 hours after exposure to 15 micrograms/ml of ceftibuten . The ceftibuten PAE was not affected by serum . The ceftibuten PAE was prolonged by exposure to a sub-minimum inhibitory concentration concentration of ceftibuten as would occur in the clinical situation . The PAE of ceftibuten was not affected by the copresence of erythromycin as would occur when treating infections in which atypical organisms are suspected . There was no correlation between bacterial reduction in colony-forming units and the duration of PAE . A level of 6 micrograms/ml of ceftibuten had a similar bacterial killing activity compared with a 6-hour exposure to 15 micrograms/ml of ceftibuten against S . pneumoniae, H . influenzae and M . catarrhalis . This study suggests that ceftibuten can be administered orally, once daily in an adult dose of 400 mg or a pediatric dose of 9 mg/kg, to treat respiratory infections caused by the most common pathogens.

Lab Anim, 1995 Jul, 29(3), 307 - 13
An enzyme-linked immunosorbent assay (ELISA) for monitoring rodent colonies for Pasteurella pneumotropica antibodies; Boot R et al.; An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of Pasteurella pneumotropica antibodies in the sera of rats, mice, hamsters and Mastomys . P . pneumotropica from mice and rats showed cross-reactivity . The ELISA using P . pneumotropica NCTC 8284 detected more infected animals than selective culture in groups of rodents from which P . pneumotropica, Haemophilus sp and/or Actinobacillus sp were cultured . Cross reactivity between P . pneumotropica NCTC 8284 and haemophilus and actinobacillus isolates were not studied.

Jpn J Antibiot, 1995 Jul, 48(7), 949 - 59
{Clinical and bacteriological effects of cefetamet pivoxil against community-acquired respiratory tract infections}; Shimada J et al.; We investigated clinical and bacteriological effects of cefetamet pivoxil (CEMT-PI) in community-acquired respiratory tract infections and obtained the following findings . 1 . Of the 420 respiratory tract infection cases that were treated with CEMT-PI according to a same protocol at a total of 42 institutions in Tokyo, Kanagawa-ken, Saitama-ken and Chiba-ken from February to the beginning of April 1994, 359 cases in which clinical evaluations were considered possible were selected as the subjects of the clinical study . Regarding genders of patients, slightly more females (56.3%) than males were included . Diagnoses given to these patients included laryngopharygealitis (60.7%), tonsillitis (14.2%) and acute bronchitis (13.6) . Outpatients accounted for 94.4% of the subjects . 2 . For the bacteriological study, a written material describing the method of collecting specimens, storage and transport in detail was distributed to the above mentioned institutions . The isolation and identification of suspected causative bacteria, determination of minimum inhibitory concentrations (MIC) and investigation of beta-lactamase production were conducted all together . Suspected causative bacteria were detected from 238 (66.3%) out of the 359 cases . They included 85 strains of Haemophilus influenzae, 76 strains of Streptococcus pneumoniae, 20 strains of Streptococcus pyogenes and 17 strains of Moraxella subgenus Branhamella catarrhalis . 3 . Clinical efficacy rates (the ratio of those excellent+good) among those who were treated with 1 CEMT-PI tablet (194 mg, titer) twice a day was 76.5% and among those who were given 2 tablets twice a day was 87.4% . The improvement rate of the latter was higher at a significant level of P < 0.05 . 4 . The clinical efficacies classified by suspected causative bacteria (single bacterium) were 93.3% against M.(B.) catarrhalis, 91.7% against beta-streptococci, 87.1% against H . influenzae and 78.4% against S . pneumoniae, etc . Though 7 (9.2%) of the 76 strains of S . pneumoniae were benzylpenicillin (PCG)-insensitive S . pneumoniae (PISP), the bacteriological efficacy was assessed either excellent or good in all of the 7 patients from whom PISP were detected . The clinical efficacy was assessed 100.0% in those from which a plural number of bacteria were detected . The 13 cases from which small numbers of Staphylococcus aureus was detected with other bacterium were also included in these cases.

Kansenshogaku Zasshi, 1995 Jul, 69(7), 777 - 84
{Haemophilus aphrophilus isolated from the blood of a patient with infective endocarditis}; Yoshioka I et al.; On July 1994, a 62-year-old female, having a history of mitral regurgitation, was admitted because of high fever, hematuria and conjunctival petechiae . She was diagnosed as having infective endocarditis with mitral valve vegetation proved by ultrasonic cardiography . The gram negative rods were isolated from blood cultures performed five times, performed prior to the administration of antibiotics . The isolates were identified as strains of H . aphrophilus . After two days of treatment with PCG (12 million units/day), the organism became undetectable from the blood . Since the minimal inhibitory concentrations (MICs) of PCG and ABPC were ranged between 0.06-2.0 micrograms/ml and 0.06-0.5 microgram/ml, respectively, ABPC was selected as a first choice antibiotic instead of PCG . ABPC was given 12 g/day for the first 3 days, then 6 g/day for 28 days, followed by 3 g/day for 7 days . The patient recovered and was discharged after the 55 hospital days . H . aphrophilus grew on BTB lactose agar, chocolate agar and sheep blood agar, but failed to grow on MacConkey agar . H . aphrophilus produced smooth transparent nonhaemolytic micro colonies after 48 hours on sheep blood agar and chocolate agar plates . Atmosphere with 5% CO2 failed to enhance their growth . All the five strains of H . aphrophilus isolated, required neither factors V nor X . Positive synthesis of porphyrin from delta-aminolevlinic acid confirmed their ability to grow without X factor . For the correct identification of H . aphrophilus strains, fermentation test of glucose, lactose, maltose and sucrose in either phenol red broth or CTA medium are necessary.(ABSTRACT TRUNCATED AT 250 WORDS)

Chemotherapy, 1995 Jul-Aug, 41(4), 316 - 22
Clinical experience with ceftriaxone treatment in the neonate; Van Reempts PJ et al.; The safety of ceftriaxone has been evaluated in 80 neonates who were treated empirically for suspected infection with either ceftriaxone and ampicillin (group A, age 0-72 h) or ceftriaxone and vancomycin (group B, age greater than 72 h) . Within 48 h after birth 2 group A patients died from sepsis (Haemophilus influenzae, Streptococcus pneumoniae, 1 case each); 1 group B patient died from sepsis (Pseudomonas aeruginosa) . All bacterial isolates from group A patients were susceptible to ceftriaxone, but in 4 of the 8 group B patients with positive cultures a change in antibiotic therapy was required . Eosinophilia, thrombocytosis and an increase in serum alkaline phosphatases were observed in a limited number of patients during and after discontinuation of treatment . Direct hyperbilirubinemia ( > 2 mg/dl) occurred in 2 cases during treatment . Gallbladder sludge was sonographically diagnosed in 6 patients, but disappeared within 2 weeks after detection . One neonate had exanthema . Nurses rated ease of administration as very good . Ceftriaxone appears to be an interesting alternative in the empiric antibiotic treatment in the early neonatal period.

Clin Microbiol Rev, 1995 Jul, 8(3), 357 - 75
Chancroid and Haemophilus ducreyi: an update; Trees DL et al.; Haemophilus ducreyi is a fastidious gram-negative bacillus that causes the sexually transmitted infection chancroid . Chancroid is a major genital ulcerative disease in Africa, Southeast Asia, the Caribbean, and Latin America and is of increasing concern in the United States . Genital ulcerative disease and chancroid in particular have been associated with facilitating the transmission of human immunodeficiency virus . The diagnosis of chancroid based on the clinical appearance of the genital lesion or on the isolation of H . ducreyi on selective medium is relatively insensitive . However, recent advances in nonculture diagnostic tests have enhanced our ability to diagnose chancroid . There has been renewed interest in understanding the pathogenesis of H . ducreyi . In vitro and in vivo models have been developed to help identify important virulence determinants . Through the use of biochemical and molecular techniques, macromolecular components that may be important in virulence have been identified.

Sex Transm Dis, 1995 Jul-Aug, 22(4), 217 - 20
Incision and drainage versus aspiration of fluctuant buboes in the emergency department during an epidemic of chancroid; Ernst AA et al.; GOAL OF THIS STUDY: To compare outcomes of incision and drainage with packing to needle aspiration of purulent buboes in patients with presumed chancroid . The safety and efficacy of incision and drainage were compared with that of needle aspiration . STUDY DESIGN AND METHODS: The study was a prospective randomized, nonblinded clinical trial carried out from April 1992, to January 1994 in an inner city emergency department . A consecutive sample of 27 adults with fluctuant inguinal buboes presumed or proven to be chancroid or lymphogranuloma venereum were eligible for inclusion . Patients were excluded if they were believed to have buboes secondary to disease other than chancroid . After informed consent was obtained, patients had buboes drained by needle aspiration or by incision and drainage with packing according to a random numbers table . RESULTS: Twenty-seven patients were included in the study (22 men and 5 women), with 12 randomized to incision and drainage and 15 to aspiration . The mean age was 35.7 +/- 13 years . Genital ulcer or bubo pus cultures for Haemophilus ducreyi were positive in seven patients, negative in 15 patients, and not done in five patients . Follow-up was obtained for 23 (85%) patients, 11 of whom had incision and drainage and 12 of whom had aspirations . No adverse effects were reported in either group . CONCLUSION: Incision and drainage is an effective method for treating fluctuant buboes and may be preferable to traditional needle aspiration considering the frequency of required re-aspirations in the study patients . Limitations of this study include lack of complete laboratory testing and lack of follow-up of all patients.

Mol Microbiol, 1995 Jul, 17(1), 123 - 35
SepA, the major extracellular protein of Shigella flexneri: autonomous secretion and involvement in tissue invasion; Benjelloun-Touimi Z et al.; In addition to Ipa proteins and IcsA, which are involved in entry into epithelial cells and intercellular spread, respectively, Shigella secretes a 110 kDa protein, designated SepA . We report the identification, cloning, and nucleotide sequence determination of the sepA gene, analysis of SepA secretion, and construction and characterization of a sepA mutant . The sepA gene is carried by the virulence plasmid and codes for a 150 kDa precursor . Upon secretion, which does not involve accessory proteins encoded by the virulence plasmid, the precursor is converted to a mature protein of 110 kDa by two cleavages removing an N-terminal signal sequence and a C-terminal fragment . Extensive similarities were detected between the sequence of the first 500 residues of mature SepA and the N-terminal region of IgA1 proteases from Neisseria gonorrhoeae and Haemophilus influenzae, the Tsh haemagglutinin of an avian pathogenic Escherichia coli, and the Hap protein involved in adhesion and penetration of H . influenzae . The C-terminal domain of the SepA precursor, which is not present in the secreted protein, exhibits sequence similarity with pertactin of Bordetella pertussis and the ring-forming protein of Helicobacter mustelae . Construction and phenotypic characterization of a sepA mutant indicated that SepA is required neither for entry into cultured epithelial cells nor for intercellular dissemination . However, in the rabbit ligated ileal loop model, the sepA mutant exhibited an attenuated virulence, which suggests that SepA might play a role in tissue invasion.

Rev Prat, 1995 Jun 15, 45(12), 1500 - 7
{Vaccination in children}; Mallet E; New vaccines, anti-Haemophilus b and anti-hepatitis B, are now available in clinical practice . Specific recommendations have led to modifying the calendar . Practitioner has to explain these new vaccines to the families because they are not compulsory . Combined vaccines (like five valence) is useful to improve compliance . In the near future, new vaccines like the well tolerated acellular pertussis, and new combined vaccines will be available.

Rev Prat, 1995 Jun 15, 45(12), 1484 - 7
{Polysaccharide subunit vaccines}; Reinert P; The polysaccharide capsules of many germs (pneumococcus, meningococcus, Haemophilus b, salmonella, etc.) can induce synthesis of protective antibodies in man . The development of many vaccines has been based on this phenomenon . Unfortunately, the polysaccharide antigens cannot trigger an immune response before the age of 2 years . In addition, they do not induce an anamnestic response, as do protein antigens . Great progress has been made by conjugating these polysaccharide antigens with protein antigens (tetanus and diphtheria antigens in particular) . The resulting novel antigen induces a thymus-dependent response in man, leading to immunogenicity from the first weeks of life, and to a higher and more durable antibody response . On subsequent administration, such conjugated vaccines provoke a booster effect . In practice, the anti-Haemophilus b conjugated vaccine has obtained spectacular results in prevention of meningitis, with an effectiveness close to 100% . Results of the first trials of conjugated anti-pneumococcal vaccines suggest that they may constitute an effective new means of combating the emerging pneumococcal strains resistant to penicillin.

FEMS Microbiol Lett, 1995 Jun 15, 129(2-3), 221 - 4
Cytotoxin production in 100 strains of Haemophilus ducreyi from different geographic locations; Purven M et al.; One-hundred strains of Haemophilus ducreyi, representing isolates from different parts of the world, including the reference strains, were obtained from different collections and characterized with special reference to cytotoxin production in vitro . The cytotoxic activity on cultured epithelial cells (HEp-2) was examined with two methods . The activity in bacterial sonicates was tested on freshly trypsinated cells and strains manifesting little or no cytotoxic activity in sonicates were investigated using attached living bacteria on HEp-2 cell-monolayers . Sonicates from the majority of the H . ducreyi strains (89%) produced significant cytotoxic effects on HEp-2 cells . The reciprocal cytotoxic titers of the sonicates ranged from 2.4 x 10(2) to 5.3 x 10(5) . Sonicates of 11 strains had low cytotoxic titers (< or = 1:3 to 1:81), eight of those originating from Asia and three from Africa . These 11 strains caused no damage to the cell monolayer, indicating that the 11 strains produce little or no cytotoxic activity in vitro . In summary, the majority of H . ducreyi isolates produce cytotoxic activity, which support the hypothesis that the cytotoxin may be an important virulence factor of this species.

Pediatr Pol, 1995 Jun, 70(6), 507 - 9
{Bacterial meningitis caused by the strains of Haemophilus influenzae type B resistant to most penicillins and cephalosporins}; Konior R; In recent years bacterial meningitis caused by Haemophilus influenzae appears to have occurred more frequently in Poland . In this paper the author presents three specific cases treated in one typical month at the Department of Neuroinfections at John Paul the II Hospital, Krakow . Each of the cases in question proved difficulty to treat because the strains of bacteria encountered were resistant to the most of penicilins and cephalosporins.

Pathol Biol (Paris), 1995 Jun, 43(6), 515 - 23
{Azithromycin, pharmacodynamic evaluation in animal models}; Lagrange PH; Several experimental models have been used in order to evaluate the in vivo efficacy of azithromycin against numerous human pathogenic bacteria and parasites, including comparison between azithromycin and other antibiotics belonging or not to the macrolide family . Using the experimental models, three major objectives can be distinguished: the comparative studies of the efficacy dose 50 (ED50) of azithromycin compared to other orally given antibiotics, the azithromycin efficacy in animal infected with intracellular multiplying micro-organisms, and the demonstration of the specific azithromycin accumulation in tissues in direct relationship with the local recruitment of phagocytic cells at the infectious foci . The ED50 of azithromycin has been compared with those of erythromycin or cefaclor in varying acute murine infections . Evidence was given of a similar efficacy for the three tested antibiotics . Nevertheless a marked advantage for azithromycin was observed in experimental local infections and with infections due to Gram-negative bacteria (Haemophilus influenzae, Branhamella catarrhalis) . The second objective was to confirm in vivo the preferential efficacy of azithromycin in models using intracellular multiplying microorganisms, due to its great capacity to accumulate inside of professional phagocytes . Several models have been used, such as those performed with Listeria monocytogenes, Legionella pneumophila, S . typhimurium, Brucella melitensis, M . avium and C . trachomatis.(ABSTRACT TRUNCATED AT 250 WORDS)

FEMS Microbiol Lett, 1995 Jun 1, 129(1), 75 - 81
Cloning and characterization of a gene from Pasteurella haemolytica A1 involved in lipopolysaccharide biosynthesis; Potter MD et al.; A Pasteurella haemolytica A1 gene involved in the biosynthesis of a moiety on the core of the lipopolysaccharide molecule has been cloned and characterized . Escherichia coli clones which carry this gene showed an alteration of its lipopolysaccharide migration profile on tricine SDS-PAGE and exhibited resistance to the core-specific phage U3 . In addition, lipopolysaccharide extracted from the E . coli clones was recognized by an anti-corespecific antiserum, but not by antiserum specific for the O antigen of P . haemolytica A1 lipopolysaccharide . Nucleotide sequence analysis of the cloned DNA identified an open reading frame (lpsA) coding for a protein of 263 amino acids which showed significant homology with a Haemophilus influenzae type b lipooligosaccharide biosynthesis gene . PCR amplification of genomic DNA, using primers based on the P . haemolytica A1 lpsA sequence, yielded products from only the A biotypes of P . haemolytica.

Epidemiol Infect, 1995 Jun, 114(3), 451 - 63
The epidemiology of Haemophilus influenzae type b disease in the Republic of Ireland; Fogarty J et al.; A 2-year case-control study was conducted to describe the epidemiology of Haemophilus influenzae type b (Hib) and investigate Hib disease risk factors in the Republic of Ireland . Between October 1991 and September 1993 . 149 laboratory confirmed incident cases were matched with community controls . Annual Hib disease incidence was 25.4 per 100,000 children under 5 years, with peak incidence (65.8 per 100,000) in the 6-11 months age-group . Meningitis was the predominant clincial condition . Twenty-four (16.1%) isolates were resistant to ampicillin . Creche or day-care attendance and the presence of chronic illness emerged as risk factors for Hib disease . Empirical first line treatment for suspected Hib infection warrants alternatives to ampicillin such as cefotaxime . Completed immunization with Hib conjugate vaccine by 6 months of age is required for maximum disease prevention . Until all children are receiving Hib vaccine on schedule, those who are creche or day-care attendees and those with chronic illness should be prioritized for timely immunization.

Curr Microbiol, 1995 Jun, 30(6), 345 - 9
A rapid dot immunoassay for detecting the Brazilian purpuric fever clone of Haemophilus influenzae biogroup aegyptius with a "flow through" device; Ajello GW et al.; Brazilian purpuric fever (BPF) is a highly fatal pediatric disease that may follow an episode of purulent conjunctivitis caused by a virulent clone of Haemophilus influenzae biogroup aegyptius (Hae) . Oral rifampin prophylaxis, by eliminating carriage of the BPF clone in children with conjunctivitis, may prevent onset of the systemic disease . A test to detect the BPF clone directly from eye swabs could identify those in need of prophylaxis . This is a preliminary report of a rapid dot immunoassay performed on a "flow-through" cartridge that was developed for use under field conditions . The test is based upon recognition of a unique epitope of the 25-kDa pilin protein on the surface of BPF clone cells by a monoclonal antibody . With 36 laboratory-maintained cultures of Hae (15 clone isolates and 21 others), sensitivity of the assay was 67% and specificity was 95% . When fimbrial-enriched (25-kDa+) phenotypes of five false-negative clone strains were prepared for use as test antigens, sensitivity rose to 100% . Evaluation of the immunoassay under field conditions is necessary to prove its efficacy.

J Bacteriol, 1995 Jun, 177(12), 3613 - 5
Ferrochelatase activity and protoporphyrin IX utilization in Haemophilus influenzae; Loeb MR; Previous research showed that the heme-requiring human pathogen Haemophilus influenzae lacks the first six of the seven enzymes required for heme synthesis, starting with the precursor, 5-amino levulinic acid . In this study, I demonstrated either directly or by reasonable inference that all 57 strains of H . influenzae examined, including 2 unable to grow on protoporphyrin IX, possess ferrochelatase, which catalyzes heme formation by insertion of Fe2+ into the protoporphyrin IX nucleus and which is the last enzyme in the heme synthetic pathway . Further, I showed that this enzyme can also function in the reverse direction, releasing Fe2+ from heme.

J Bacteriol, 1995 Jun, 177(11), 3235 - 40
DNA sequence and characterization of Haemophilus influenzae dprA+, a gene required for chromosomal but not plasmid DNA transformation; Karudapuram S et al.; Natural genetic transformation in Haemophilus influenzae involves DNA binding, uptake, translocation, and recombination . In this study, we cloned and sequenced a 3.8-kbp H . influenzae DNA segment capable of complementing in trans the transformation defect of an H . influenzae strain carrying the tfo-37 mutation . We used subcloning, deletion analysis, and in vivo protein labeling experiments to more precisely define the gene required for efficient DNA transformation on the cloned DNA . A novel gene, which we called dprA+, was shown to encode a 41.6-kDa polypeptide that was required for efficient chromosomal but not plasmid DNA transformation . Analysis of the deduced amino acid sequence of DprA suggested that it may be an inner membrane protein, which is consistent with its apparent role in DNA processing during transformation . Four other open reading frames (ORFs) on the cloned DNA segment were identified . Two ORFs were homologous to the phosphofructokinase A (pfkA) and alpha-isopropyl malate synthase (leuA) genes of Escherichia coli and Salmonella typhimurium, respectively . Homologs for the two other ORFs could not be identified.

J Bacteriol, 1995 Jun, 177(11), 3111 - 9
Biochemical and genetic analyses of a catalase from the anaerobic bacterium Bacteroides fragilis; Rocha ER et al.; A single catalase enzyme was produced by the anaerobic bacterium Bacteroides fragilis when cultures at late log phase were shifted to aerobic conditions . In anaerobic conditions, catalase activity was detected in stationary-phase cultures, indicating that not only oxygen exposure but also starvation may affect the production of this antioxidant enzyme . The purified enzyme showed a peroxidatic activity when pyrogallol was used as an electron donor . It is a hemoprotein containing one heme molecule per holomer and has an estimated molecular weight of 124,000 to 130,000 . The catalase gene was cloned by screening a B . fragilis library for complementation of catalase activity in an Escherichia coli catalase mutant (katE katG) strain . The cloned gene, designated katB, encoded a catalase enzyme with electrophoretic mobility identical to that of the purified protein from the B . fragilis parental strain . The nucleotide sequence of katB revealed a 1,461-bp open reading frame for a protein with 486 amino acids and a predicted molecular weight of 55,905 . This result was very close to the 60,000 Da determined by denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified catalase and indicates that the native enzyme is composed of two identical subunits . The N-terminal amino acid sequence of the purified catalase obtained by Edman degradation confirmed that it is a product of katB . The amino acid sequence of KatB showed high similarity to Haemophilus influenzae HktE (71.6% identity, 66% nucleotide identity), as well as to gram-positive bacterial and mammalian catalases . No similarities to bacterial catalase-peroxidase-type enzymes were found . The active-site residues, proximal and distal hemebinding ligands, and NADPH-binding residues of the bovine liver catalase-type enzyme were highly conserved in B . fragilis KatB.

Infect Immun, 1995 Jun, 63(6), 2327 - 33
Affinity, conservation, and surface exposure of hemopexin-binding proteins in Haemophilus influenzae; Wong JC et al.; Haemophilus influenzae can acquire heme from hemopexin for use as a source of both essential porphyrin and iron . In classical ligand-binding studies, we observed time-dependent, saturable, and displaceable binding of human 125I-labelled hemopexin to intact cells of H . influenzae type b (Hib) strain 760705 grown in an iron-restricted medium . From these experiments, which demonstrate that hemopexin associates with a single class of binding site, the affinities (Kds) and receptor numbers were calculated for heme-hemopexin (Kd, 205 nM; 3,200 receptors per cell) and apohemopexin (Kd, 392 nM; 4,400 receptors per cell) . Thus, Hib expresses a specific hemopexin receptor which shows some preference for the heme-protein complex . Affinity chromatography on hemopexin-Sepharose 4B of detergent-solubilized membranes from Hib strain 760705 results in the copurification of three proteins with molecular masses of 57, 38, and 29 kDa . Trypsinization of whole cells of Hib 760705 abolishes hemopexin binding and correlates with the disappearance of the 57-kDa hemopexin-binding protein and appearance of a 52-kDa species which does not bind either hemopexin in ligand blot assays or a monoclonal antibody (MAbT11-30) raised against the 57-kDa protein . From immunoblotting assays and NH2-terminal amino acid sequence analysis, the 38-kDa protein isolated following hemopexin affinity chromatography was identified as the porin protein P2 . These data, taken together with the receptor-binding studies which support a single class of hemopexin-binding site, suggest that P2 and the 29-kDa protein function as accessory proteins to the 57-kDa hemopexin-binding protein to facilitate the uptake of heme from receptor-bound hemopexin . To determine whether hemopexin binding and the 57-kDa protein are conserved in Haemophilus strains, whole-cell dot blots and immunoblots of the outer membrane proteins prepared from strains belonging to each of 21 different Hib outer membrane protein subtypes, six nontypeable strains, and five Haemophilus parainfluenzae strains were probed with either hemopexin or MAbT11-30 . Only the H . parainfluenzae strains which lack the 57-kDa protein do not bind hemopexin . Since H . influenzae has also been shown to produce a soluble 100-kDa hemopexin-binding protein, cell-free culture supernatants were also examined for the presence of this protein . Apart from Hib 760705 and H . parainfluenzae, the 100-kDa hemopexin-binding protein was detected in all the other Haemophilus strains . The abilities of Hib 760705 to both bind and acquire heme from hemopexin without expressing a 100-kDa soluble hemopexin-binding protein show that in strain 760705, this 100-kDa protein is not essential for the utilization of heme from hemopexin.

Infect Immun, 1995 Jun, 63(6), 2317 - 22
Human microvascular endothelial tissue culture cell model for studying pathogenesis of Brazilian purpuric fever; Quinn FD et al.; Brazilian purpuric fever (BPF) is a fulminant pediatric disease characterized by fever, with rapid progression to purpura, hypotensive shock, and death . All known BPF cases have been caused by three clones of Haemophilus influenzae biogroup aegyptius and have occurred in either Brazil or Australia . Using an immortalized line of human vascular endothelial cells, we developed an in vitro assay that identifies all known BPF-causing H . influenzae biogroup aegyptius strains (R . S . Weyant, F . D . Quinn, E . A . Utt, M . Worley, V . G . George, F . J . Candal, and E . W . Ades, J . Infect . Dis . 169:430-433, 1994) . With multiplicities of infection (MOIs) as low as one bacterium per 1,000 tissue culture cells, BPF-associated strains produce a unique cytotoxic effect in which the tissue culture cells detach and aggregate in large floating masses after 48 h of incubation . In this study, using a BPF-associated strain and a non-BPF-associated control, we demonstrated that strains which produce the cytotoxic phenotype were able to replicate intracellularly whereas non-BPF-associated strains, with MOIs of > or = 1,000 did not replicate and did not produce the phenotype . We also showed that this phenotype is not caused by the activity of an endotoxin or the release of some other compound from the bacterial cell, since neither gamma irradiation-killed whole BPF clone bacteria nor bacterial cell fractions at MOIs of > 1,000 produced the cytotoxic effect . Furthermore, bacteria in numbers equal to MOIs of > 1,000 treated with chloramphenicol did not produce the cytotoxic phenotype, suggesting a requirement for bacterial protein synthesis . In addition, viable bacteria separated from the tissue culture monolayer by a 0.2-micron-pore-size membrane also failed to produce the phenotype . The ability of the bacterium to invade, replicate, and produce the phenotype appears to be primarily parasite directed since phagocytosis, pinocytosis, and eukaryotic protein synthesis inhibitors, including cycloheximide, cytochalasin D, and methylamine, had no effect on the ability of the bacterium to invade and cause a cytotoxic response . Understanding the basic mechanisms involved in this tissue-destructive process should enhance our knowledge of the general pathogenesis of BPF.

Infect Immun, 1995 Jun, 63(6), 2194 - 200
Characterization of the hgbA locus encoding a hemoglobin receptor from Haemophilus ducreyi; Elkins C et al.; Haemophilus ducreyi can bind hemoglobin and use it as a source of heme, for which it has an obligate requirement . We previously identified and purified HgbA, a hemoglobin-binding outer membrane protein from H . ducreyi . In this report, we describe the molecular cloning, expression, DNA sequence, and mutagenesis of the structural gene for HgbA, hgbA . H . ducreyi and recombinant Escherichia coli expressing hgbA bound {125I}hemoglobin, establishing HgbA as a receptor . Insertions or deletions in the cloned hgbA gene abolished expression of HgbA and hemoglobin binding in E . coli . Mutagenesis of H . ducreyi by allelic exchange of insertions into hgbA abolished its ability to bind {125I}hemoglobin or utilize hemoglobin as a source of heme . The deduced protein sequence was similar to those of the TonB-dependent family of outer membrane receptors . The most similar member was HutA (heme receptor) from Vibrio cholerae . Tbp1 and Lbp1 (transferrin and lactoferrin receptors, respectively, from pathogenic Neisseria spp.) also showed very significant homology . Thus, by characterizing the hgbA locus, this work elucidates a potentially important role of HgbA in obtaining heme and/or iron from the host.

Am J Respir Crit Care Med, 1995 Jun, 151(6), 2094 - 9; discussion 2099-100
Virulence factors in the colonization and persistence of bacteria in the airways; van Alphen L et al.; Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis, Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa are commonly isolated from sputum specimens of patients with lower respiratory tract infections . Haemophilus influenzae, S . pneumoniae, and M . catarrhalis have several pathogenic properties in common . These bacteria are able to interact with mucus, to exert ciliotoxic activity, to adhere to bronchial epithelial cells, and to invade airway epithelium . Haemophilus influenzae and S . pneumoniae strains persist for many months in the respiratory tract of patients with chronic obstructive pulmonary disease (COPD), despite the specific antibodies present in serum and sputum against the persistent strain . Especially during exacerbations persistent strains with changes in their antigenic composition are isolated . Among H . influenzae strains, the antigenic characteristics of the outer membrane protein composition vary . Variation in S . pneumoniae occurs in capsular polysaccharides, the major immunogens of this bacterium . Such variations affect the efficacy of the antibody-mediated defense mechanisms against the bacteria . Between exacerbations, particularly H . influenzae, S . pneumoniae strains are recovered from the sputum of patients with COPD . Recovery may continue for periods up to 2 yr, although not continuously . Besides ineffective antibody-mediated defense mechanisms, it is likely that hiding of the bacteria in tissue contributes to the persistence of these bacteria in patients with COPD.

Am J Respir Crit Care Med, 1995 Jun, 151(6), 2081 - 6; discussion 2086-7
The role of humoral mucosal immunity in the induction and maintenance of chronic airway infections; Brandtzaeg P; The respiratory mucosa is protected primarily by a secretory immune system that is under complex and only partly understood immunoregulatory control . Secretory immunoglobulins (SIgA and SIgM) protect the mucosal surface by immune exclusion of antigens . However, the fact that most IgA produced in the respiratory tract belongs to the IgA1 subclass renders SIgA in this region susceptible to IgA-specific proteases produced by Haemophilus influenzae, Streptococcus pneumonia, and Neisseria meningitidis . Immunoglobulin G can also perform immune exclusion at respiratory surfaces but, like IgE, it reaches the secretions merely by passive diffusion . The phlogistic properties of antibodies belonging to these classes explain their potential involvement in maintaining mucosal inflammation . In patients with selective IgA deficiency, SIgA is lacking and is not regularly compensated for satisfactorily by SIgM . In such patients unexplained immunoregulatory mechanisms, perhaps involving the local microbiota, give rise to a large number of IgD-producing cells in the upper respiratory tract . Immunoglobulin D cannot act as a secretory antibody and might block the protective properties of IgG; this could explain why these patients are particularly prone to recurrent infections . Our observations show that there are large individual variations in the mucosal immune system with regard to humoral immunity in the upper respiratory tract.

Pediatrics, 1995 Jun, 95(6), 815 - 22
Neonatal immunization: response to Haemophilus influenzae type b-tetanus toxoid conjugate vaccine; Kurikka S et al.; OBJECTIVE . To study the immunogenicity and tolerability of Haemophilus influenzae type b (Hib) conjugate vaccine administered in the neonatal period . DESIGN: Hib capsular polysaccharide (PS)-tetanus toxoid conjugate vaccine (PRP-T) was given to 120 neonates at 2 days of age, followed by PRP-T or the Hib PS vaccine at 4 months and a PRP-T booster at 14 months . Their anti-Hib PS concentrations were compared with those in children receiving PRP-T at 2 and 4 months or at 4 months . RESULTS . No serious adverse reactions were noted . The geometric mean concentration of anti-Hib PS at the age of 2 days was 0.34 micrograms/mL and at 4 months was 0.12 micrograms/mL . This was significantly more than the concentration in unimmunized infants at this age and 3.5 times more than expected, taking into account the natural decay of transplacentally acquired antibodies . Such a response was not seen in infants with a high (greater than 3.0 micrograms/mL) neonatal antibody concentration . The PRP-T vaccine given at 4 months elicited an antibody response in all infants and Hib PS in 62%, indicating immunologic priming . At 14 months, a higher percentage of the infants who had received PRP-T at 2 days and 4 months than of those who had received PRP-T at 4 months only had anti-Hib PS concentrations greater than 0.15 micrograms/mL . All infants responded well to the booster at 14 months . There was no evidence of immunologic tolerance . CONCLUSIONS . Neonatal immunization with PRP-T was safe and well tolerated in Finnish infants, and it would be worthwhile to further study its effects in higher risk populations.

Ann Trop Paediatr, 1995 Jun, 15(2), 179 - 83
Inappropriate secretion of antidiuretic hormone in acute bacterial meningitis; Patwari AK et al.; Sixty children aged from 1 month to 12 years (mean (SD) 3.18 (3.49) years) with acute bacterial meningitis were studied for the incidence, clinical manifestations and outcome of the inappropriate secretion of antidiuretic hormone syndrome (SIADH) . Serum sodium levels and osmolality of serum and urine were estimated on admission and on days 3 and 10 . SIADH was diagnosed in 22 out of 60 cases (36.7%) on admission and in six of 48 cases (12.5%) on day 3 . Hyponatraemia without SIADH, attributed to vomiting and fever, was detected in seven cases (11.7%) . Serum sodium levels returned to normal within 48 hours in these cases . Serum osmolality and sodium levels took longer to return to normal values in patients with SIADH . However, none of the cases showed any evidence of SIADH on the 10th day . A significant correlation with SIADH was observed in cases with evidence of severe meningeal inflammation (p < 0.001) . The incidence of SIADH was highest with Streptococcus pneumoniae (75%), followed by Haemophilus influenzae (57.1%) . Overall mortality was 26.7%, and mortality was significantly higher (p < 0.001) in cases with SIADH, all of whom died during the 1st 72 hours . Ten out of 22 cases (45.4%) with SIADH who survived beyond the 1st 72 hours had an uneventful course even though all of them had biochemical evidence of SIADH on the 3rd day . Mortality was quite high also in children with severe malnutrition (75%) and in those with S . pneumoniae as the aetiological organism (75%).

Ann Trop Paediatr, 1995 Jun, 15(2), 173 - 7
A bacteriological study in hospitalized children with pneumonia; Bahl R et al.; A total of 110 consecutive hospitalized children with severe lower respirator tract infection were studied with the aim of determining the main bacterial pathogens responsible . Of these, 57 were classified as severe pneumonia and 53 as very severe pneumonia . Streptococcus pneumoniae was the most common organism identified in 24.6% and 32.1% of cases of severe and very severe pneumonia, respectively, followed by Haemophilus influenzae type b, which was seen in 17.5% and 20.8%, respectively . The highest diagnostic yield was with the latex particle agglutination test on serum and urine . Blood culture was positive in only ten of the 110 children . No difference was found between the aetiological agents identified in severe and very severe cases of pneumonia . Therefore, the use of different parenteral antibiotics for two clinically defined groups of pneumonia, as recommended by WHO in their standard case management guidelines for the ARI control programme, does not seem necessary.

Infect Dis Clin North Am, 1995 Jun, 9(2), 325 - 33
Immunization . Past successes, future challenges; Grossman M; Global eradication of smallpox, elimination of poliomyelitis in the Western Hemisphere, and a 90% decrease in Haemophilus influenzae meningitis in the Western Hemisphere mark some of the achievements of immunization programs around the world . Yet many serious problems continue to exist, particularly in the developing world . Advances in biotechnology and the renewed interest by industry, governments, and the World Health Organization in vaccine development and delivery in the field provide an exciting prospect for major advances in immunization in the next few decades.

Pediatr Infect Dis J, 1995 Jun, 14(6), 517 - 21
Acquisition of IgG serum antibodies against two Bordetella antigens (filamentous hemagglutinin and pertactin) in children with no symptoms of pertussis; Isacson J et al.; To study the specificity of serum antibodies against filamentous hemagglutinin (FHA) and pertactin for infection with Bordetella pertussis, we followed the acquisition of IgG serum antibodies against these 2 surface proteins of the organism in children who had been vaccinated with a monocomponent pertussis toxoid vaccine and who had experienced no symptoms of pertussis . Antibodies were estimated with enzyme-linked immunosorbent assay . In Part 1 of our study 5 consecutive samples obtained between 3 and 36 months of age from 71 children were available . Most had maternally derived antibodies to FHA (70 of 71) and pertactin (51 of 71) in the 3-month sera which declined in the subsequent sera . From about 1 year of age there were small but significant increases in antibodies against both antigens . At 3 years of age 71 of 71 had antibodies to FHA and 58 of 71 had antibodies to pertactin . In Part 2 of our study sera from 109 three-year old children were available . The 12 children with a history of family exposure to pertussis had significantly higher geometric mean titers of FHA antibodies than the 97 children with no history of family exposure . The geometric mean titers of pertactin antibodies did not differ . We suggest 3 explanations for the acquisition of FHA and pertactin antibodies in children with no history of pertussis: (1) asymptomatic B . pertussis infection in vaccinated children; (2) infection with Bordetella parapertussis; (3) infection with cross-reacting antigens from other organisms, e.g., nonencapsulated Haemophilus influenzae.

Pediatr Infect Dis J, 1995 Jun, 14(6), 478 - 84
Bacterial antibody assays in the diagnosis of acute lower respiratory tract infection in children; Nohynek H et al.; Bacterial antibodies were studied in acute, intermediate and convalescent phase sera (mean duration from first to last sample 36 days) of 121 children hospitalized for acute lower respiratory tract infection . Antibody responses were observed in 45% of all cases and in 29% of the 21 children < 1 year old . A total of 15 responses to Streptococcus pneumoniae (pneumolysin), 20 to Haemophilus influenzae, 9 to Moraxella catarrhalis, 3 to chlamydiae and 8 to Mycoplasma pneumoniae were found . In 79 patients with 4 consecutive samples available, 52% of the 31 responses were measurable within 5 days from admission . Overall the responses were not associated with upper respiratory tract bacterial findings or acute otitis media . Significantly more responses were found in the 121 children with acute lower respiratory tract infection than in healthy controls (P < 0.007) . We conclude that bacterial antibody assays provide a useful tool in the study of the etiology of acute lower respiratory tract infection in young children, even if the interval between paired serum samples is short.

Int J Pediatr Otorhinolaryngol, 1995 Jun, 32 Suppl, S127 - 34
Immunological aspects of otitis media: present views on possibilities of immunoprophylaxis of acute otitis media in infants and children; Karma PH et al.; The article reviews, based on current knowledge of immunological events affecting the middle ear, the possibilities and prospects for the prevention of otitis media (OM) by immunologic measures . While pneumococcal capsular polysaccharide vaccines proved not to be effective against infant acute otitis media (AOM), pneumococcal conjugate vaccines provide good immunogenicity even in infants, and call for trials with better prospects of clinical efficacy . The other future approaches currently under development are vaccines against nontypable Haemophilus influenzae and Branhamella catarrhalis, anti-viral immunoprophylaxis, combinations of the above alternatives, or passive immunization . Also, the use of new routes or ways of immunization are under study . Furthermore, the ways to modify the present treatment practices of AOM to favour good immunologic responses in infants and children must be studied.

Laryngorhinootologie, 1995 Jun, 74(6), 355 - 60
{Cephalosporin treatment of maxillary sinusitis}; Kohler W et al.; A new oral prodrug cephalosporin, cefcanel daloxate, was compared with cefaclor, another oral cephalosporin, in the treatment of acute purulent maxillary sinusitis . Three hundred fifty-four patients with purulent sinus secretion were recruited and randomized at thirteen European ENT clinics for the double blind study . The patients were treated with either cefcanel 300 mg tablets twice daily or cefaclor 250 mg capsules three times daily for ten days . Apart from the sinus puncture and secretion aspiration, the existence of sinus pain and purulent nasal secretion were inclusion criteria . Between days 12 and 15, reaspiration of the sinus was performed and signs and symptoms noted . The latter were checked again at the final visit between days 25 and 30 . Samples of blood and urine were obtained for safety evaluation at inclusion, during and after treatment . The bacteriological culture of the purulent sinus secretion in 58.4% of patients revealed Haemophilus influenzae (37.9%) as the most prevalent pathogen followed by Streptococcus pneumonia (19.7%), while Streptococcus pyogenes, Branhamella catarrhalis, and Staphylococcus aureus amounted each to 6-7% only . The overall clinical outcome was defined as cure/improvement in 83.3% of the patients in the cefcanel group and 89.3% in the cefaclor group, respectively . No statistically significant differences with regard to efficacy were found between the two treatment groups . Adverse events, mostly related to gastrointestinal reactions, were reported by 36 patients (15.7%) in the cefcanel group and by 16 (13.4%) in the cefaclor group . Both cefcanel 300 mg twice daily and cefaclor 250 mg three times daily were highly effective with only minimal adverse events in the treatment of acute purulent maxillary sinusitis.

J Clin Microbiol, 1995 Jun, 33(6), 1665 - 7
Multicenter clinical laboratory evaluation of a beta-lactamase disk assay employing a novel chromogenic cephalosporin, S1; Doern GV et al.; S1, a new chromogenic cephalosporin (International BioClinical, Inc., Portland, Oreg.), was used to detect beta-lactamase production among a variety of commonly encountered bacteria in a four-center collaborative study . Results of an S1 disk assay were compared with those obtained by a nitrocefin-based disk procedure (Cefinase; Becton-Dickinson Microbiology Systems, Cockeysville, Md.), with repetitive testing of five quality control organisms and with individual tests of recent clinical isolates of Neisseria gonorrhoeae (162 strains), Haemophilus influenzae (162 strains), Moraxella catarrhalis (155 strains), Staphylococcus aureus (161 strains), and Bacteroides fragilis (164 strains) . The performances of the two beta-lactamase disk assays were comparable for the first three species cited above . However, the S1 assay appeared to be a more sensitive procedure than the Cefinase assay when applied to S . aureus and B . fragilis, with respect to both total numbers of positive results and length of time to a definitive positive endpointPublication Types:

Multicenter Study

What Is Nitrification?, What Is Biofilter?, What Is MIC?, What Is Genome?, What Is Amino Acid?, c, Microbiology, r, Bacteria, n, Bacteriology, a, Bacterium, r, Microbe, n, Escherichia coli, o, S. cerevisiae, i, E coli O157, n, Fermentations, c, Escherichia coli, e, Fermentations, i, Clostridia, i, Thermophiles, n, Antibiotics, n, Pseudomonas aeruginosa, c, Neisseria, o, Staphylococcus, e, Bacillus, e, Enterobacters, n, Fermentations, s, Escherichia coli, a, Ps. putida, i, Neisseria, o, Salmonella typhimurium, i, Escherichia coli, o, Sepsis

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005