Haemophilus

Nucleic Acids Res, 1998 Jun 15, 26(12), 2971 - 80
Oligonucleotide bias in Bacillus subtilis: general trends and taxonomic comparisons; Rocha EP et al.; We present a general analysis of oligonucleotide usage in the complete genome of Bacillus subtilis . Several datasets were built in order to assign various biological contexts to the biased use of words and to reveal local asymmetries in word usage that may be coupled with replication, the control of gene expression and the restriction/modification system . This analysis was complemented by cross-comparisons with the complete genomes of Escherichia coli , Haemophilus influenzae and Methanococcus jannaschii . We have observed a large number of biased oligonucleotides for words of size up to 8, throughout the datasets and species, indicating that such long strict words play an important role as biological signals . We speculate that some of them are involved in interactions with DNA and/or RNA polymerases . An extensive analysis of palindrome abundances and distributions provides the surprising result that prophage-like elements embedded in the genome exhibit a smaller avoidance of restriction sites . This may reinforce a recently proposed hypothesis of a selfish gene phenomena in the transfer of restriction/modification systems in bacteria.

An Esp Pediatr, 1998 Mar, 48(3), 277 - 82
{Bacterial meningitis . Clinical-epidemiological study . Review of 8 years (1988-1995)}; Martinez Leon M et al.; OBJECTIVE: The objective of this study was to determine the epidemiological and clinical characteristics of the bacterial meningitis assisted in the Hospital Materno-Infantil of Complejo Hospitalario Carlos Haya of Malaga . PATIENTS AND METHODS: The epidemiological, clinical, biological and therapeutical aspects are analyzed between the period of January 1988 to December 1995 . RESULTS: The number of cases was 322, with the pathogen known in 240 (74.6%) and undetermined in 82 cases (24.6%) . Meningococcal meningitis was the predominant cause with 162 cases (67.5%), followed by Haemophilus influenzae (40 cases, 16.7%) and pneumococcal meningitis (27 cases, 11.3%) . Neisseria meningitidis type B was the most common serogroup, with an increasing number of type C from 1993.

J Infect Dis, 1998 Jun, 177(6), 1608 - 13
Prevention of experimental Haemophilus ducreyi infection: a randomized, controlled clinical trial; Thornton AC et al.; Human subjects were infected with Haemophilus ducreyi . All subjects developed papules and were randomized to treatment with a single dose of azithromycin (1 g) or ciprofloxacin (500 mg) . At weekly intervals, volunteers were reinoculated with H . ducreyi, and drug concentrations were measured in peripheral blood mononuclear cells (PBMC) . When papules developed, the subjects were treated with antibiotics and dismissed from the study . Eight of the ciprofloxacin-treated subjects developed papules 1 week after the initial treatment, and the ninth subject developed disease 2 weeks after treatment . The 9 azithromycin-treated subjects developed papules 4-10 weeks (mean, 6.8) after the initial treatment (P < .001) . Azithromycin was detected in PBMC for 3-6 weeks (mean, 4) . Pre- and posttreatment lesions had histology typical of experimental chancroid or were culture positive . Azithromycin prevents experimental chancroid for nearly 2 months . These findings have implications for strategies to prevent chancroid.

J Infect Dis, 1998 Jun, 177(6), 1758 - 61
Vaccination with a Haemophilus influenzae type b conjugate vaccine reduces oropharyngeal carriage of H . influenzae type b among Gambian children; Adegbola RA et al.; The effect of a Haemophilus influenzae type b (Hib) polyribosylribitol phosphate-tetanus toxoid conjugate vaccine (Hib/PRP-T) on oropharyngeal carriage of Hib was studied during an efficacy trial in Gambian infants . Children were vaccinated with Hib/PRP-T and diphtheria-tetanus toxoids-pertussis (DTP) or DTP alone at ages 2, 3, and 4 months . Groups of 1000 children aged 1-2 years were studied each year for 4 years . Hib was detected by production of a halo on antiserum agar plates . Carriage was significantly lower among children fully vaccinated with Hib/PRP-T given with DTP (4.4%; 95% confidence interval {CI}, 3.8%-5.7%) than among children fully vaccinated with DTP alone (11.0%; 95% CI, 8.9%-13.0%) (protective effect adjusted by year = 60%; 95% CI, 44%-72%; P < .001) . Hib carriage varied by year among nonvaccinated children . Hib conjugate vaccines are likely to produce a herd protective effect in underdeveloped communities, as recorded in Europe and the United States.

Vaccine, 1998 Jan, 16(1), 109 - 13
Randomised controlled trial of combined diphtheria, tetanus, whole-cell pertussis vaccine administered in the same syringe and separately with Haemophilus influenzae type b vaccine at two, three and four months of age; Jones IG et al.; An open randomised controlled multicentre study compared the immunogenicity and reactogenicity of three vaccines given by injection at two, three and four months of age . Children (89) received Haemophilus influenzae type b (Hib) vaccine (SmithKline Beecham Biologicals {SB}) administered in the same syringe with combined diphtheria-tetanus-whole-cell pertussis (DTPw) vaccine (Evans); 75 received Hib vaccine (SB) administered as a separate injection with DTPw vaccine; 66 received Hib vaccine (Pasteur Merieux {PM}) administered as a separate injection with DTPw vaccine . All subjects in both groups receiving Hib (SB) vaccine had levels of antibodies to the Hib polysaccharide polyribosylribitol phosphate (PRP) greater than 0.15 microgram ml-1 as did 97% of those receiving Hib (PM) vaccine 1 month after administration of the final vaccine dose . Subjects in all three groups demonstrated an immunological response to pertussis, diphtheria and tetanus antigens . The geometric mean titres of the group given Hib (SB) and DTPw vaccine mixed in the same syringe were lower than the other groups . There were no apparent differences between the treatment groups in the incidence of local or systemic reactions, or serious adverse events . This study has confirmed that it is possible to halve the number of injections necessary to offer protection, with advantages to parents, children, doctors and nurses, using a combined DTPwHib vaccine and in accordance with the UK's accelerated primary immunisation schedule at two, three and four months of age.

Mol Med, 1998 Apr, 4(4), 231 - 9
T-cell epitopes in type 1 diabetes autoantigen tyrosine phosphatase IA-2: potential for mimicry with rotavirus and other environmental agents; Honeyman MC et al.; The tyrosine phosphatase IA-2 is a molecular target of pancreatic islet autoimmunity in type 1 diabetes . T-cell epitope peptides in autoantigens have potential diagnostic and therapeutic applications, and they may hold clues to environmental agents with similar sequences that could trigger or exacerbate autoimmune disease . We identified 13 epitope peptides in IA-2 by measuring peripheral blood T-cell proliferation to 68 overlapping, synthetic peptides encompassing the intracytoplasmic domain of IA-2 in six at-risk type 1 diabetes relatives selected for HLA susceptibility haplotypes . The dominant epitope, VIVMLTPLVEDGVKQC (aa 805-820), which elicited the highest T-cell responses in all at-risk relatives, has 56% identity and 100% similarity over 9 amino acids (aa) with a sequence in VP7, a major immunogenic protein of human rotavirus . Both peptides bind to HLA-DR4(*0401) and are deduced to present identical aa to the T-cell receptor . The contiguous sequence of VP7 has 75% identity and 92% similarity over 12 aa with a known T-cell epitope in glutamic acid decarboxylase (GAD), another autoantigen in type 1 diabetes . This dominant IA-2 epitope peptide also has 75-45% identity and 88-64% similarity over 8-14 aa to sequences in Dengue, cytomegalovirus, measles, hepatitis C, and canine distemper viruses, and the bacterium Haemophilus influenzae . Three other IA-2 epitope peptides are 71-100% similar over 7-12 aa to herpes, rhino-, hanta- and flaviviruses . Two others are 80-82% similar over 10-11 aa to sequences in milk, wheat, and bean proteins . Further studies should now be carried out to directly test the hypothesis that T-cell activation by rotavirus and possibly other viruses, and dietary proteins, could trigger or exacerbate beta-cell autoimmunity through molecular mimicry with IA-2 and (for rotavirus) GAD.

Lancet, 1998 May 16, 351(9114), 1472 - 6
Economisation of vaccination against Haemophilus influenzae type b: a randomised trial of immunogenicity of fractional-dose and two-dose regimens; Lagos R et al.; BACKGROUND: The cost of Haemophilus influenzae type b (Hib) conjugate vaccines has limited their use in non-industrialised countries . To identify more economical vaccination schedules, we carried out a randomised trial of the immunogenicity of alternative regimens to the standard three-dose series . METHODS: 627 Chilean infants were randomly allocated to one of four regimens with either Hib polysaccharide-tetanus toxoid conjugate vaccine (PRP-T) or Hib oligosaccharide-diphtheria mutant toxoid conjugate vaccine (PRP-CRM197), for a total of eight groups . All infants receive diphtheria-tetanus-pertussis (DTP) vaccine at ages 2, 4, and 6 months . The regimens included three full doses, three fractional doses consisting of one half or one third of the full dose, and a regimen of two full doses (at age 4 and 6 months) . The primary outcome was the proportion of infants with serum anti-polyribosylribitol phosphate (PRP, the type b capsular polysaccharide) concentrations of 0.15 microg/mL or more at age 8 months . FINDINGS: 93% (95% CI 85-98) of infants vaccinated with three full doses of PRP-T or PRP-CRM197 (95% CI 84-98) achieved anti-PRP concentrations of 0.15 microg/mL or more at age 8 months, compared with 91% (83-96) to 100% (95-100) of infants immunised with any fractional-dose regimen . Of the infants vaccinated with two doses of PRP-T or PRP-CRM197, 99% (93-100) and 87% (77-93) developed anti-PRP concentrations of 0.15 microg/mL or more, respectively . INTERPRETATION: 91% (83-96) to 100% (95-100) of infants immunised with one-half or one-third of a full dose of Hib conjugate developed protective antibody concentrations . Carrier priming with DTP may make two-dose schedules an option in some places . These alternative regimens could bring the cost of Hib vaccines within reach of countries that currently cannot afford them.

Lancet, 1998 May 16, 351(9114), 1446 - 7
Getting Hib vaccine to those who need it; Booy R; PIP: Despite the availability of an effective vaccine, Haemophilus influenzae type b (Hib) accounts for more than 3 million cases of serious illness and 700,000 deaths annually, mostly in developing countries where the cost of the vaccine is prohibitive . The World Health Organization has recommended that Hib conjugate vaccines be included in infant immunization programs, especially in countries with high disease rates . In light of evidence from a study by Rosanna Lagos et al., that 3 fractional doses or 2 full doses of vaccine may be as effective as the standard 3-dose infant regimen, research into the efficacy of a single dose in resource-poor settings would be valuable . However, it is unlikely that vaccine manufacturers will reformulate their vaccines to produce substantially cheaper products because a 66.7% reduction in antigen content is estimated to reduce costs by only 10% . Reformulation would also necessitate reapplication for a product license, which is an expensive undertaking . While there is a dearth of published material available on dose-ranging immunogenicity studies with Hib conjugate vaccines, selection of a dose reflects the inferior field vs . trial delivery conditions, and the fact that some recipients will be less immunocompetent than trial participants . The most cost-saving implication of the Lagos study may be found in the temptation to substitute a single dose for a multi-dose of the vaccine . This practice, however, would raise concerns about preservation, contamination, and vaccine failure . New competitors in the Hib market may lower the price within reach, but policymakers should understand that even a high-priced vaccine can be cost-effective as a medical intervention .

Am J Respir Crit Care Med, 1998 May, 157(5 Pt 1), 1498 - 505
Bronchial microbial patterns in severe exacerbations of chronic obstructive pulmonary disease (COPD) requiring mechanical ventilation; Soler N et al.; We carried out a comprehensive microbiological study of the upper and lower airways in patients with severe exacerbations of chronic obstructive pulmonary disease (COPD) requiring mechanical ventilation in order to describe microbial patterns and analyze their clinical significance . Quantitative cultures of tracheobronchial aspirates (TBAs), bronchoscopically retrieved protected specimen brush (PSB) and bronchoalveolar lavage fluid (BALF) at admission to the ICU and after 72 h, as well as serology for bacteria and respiratory viruses were performed . Fifty patients (mean age 68 +/- 8, 46 males) were studied prospectively . Potentially pathogenic microorganisms (PPMs) and/or a positive serology were present in 36 of 50 (72%) patients, including 12 (33%) polymicrobial cases . Only six (12%) had no pathogen in any sample in the absence of antimicrobial pretreatment . Microbial patterns corresponded to community-acquired pathogens (Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis) in 19 of 34 (56%) and to gram-negative enteric bacilli (GNEB), Pseudomonas, and Stenotrophomonas spp . in 15 of 34 (44%) of isolates . Chlamydia pneumoniae and respiratory viruses were found in 18% and 16% of investigations, respectively . Repeated investigation after 72 h in 19 patients with PPMs in the initial investigation revealed eradication of virtually all isolates of community-acquired pathogens and GNEB but persistence of three of five Pseudomonas spp . and both Stenotrophomonas spp . as well as the emergence of new GNEB, Pseudomonas and Stenotrophomonas spp . Clinical parameters neither predicted the presence of PPMs nor of GNEB and Pseudomonas/Stenotrophomonas spp . Nevertheless, severe pneumonia attributable to initially isolated pathogens occurred in two patients with severe COPD exacerbation . We conclude that pathogens were more frequently present than previously reported . The rate of GNEB and Pseudomonas/Stenotrophomonas spp . isolates was high . The presence of pathogens was clinically unpredictable . Thus, in this population of patients with severe exacerbations of COPD, it may be advisable to obtain respiratory samples and to treat according to diagnostic results . Further studies are warranted to clarify this issue.

J Int Med Res, 1998 Mar-Apr, 26(2), 66 - 75
A comparison of the efficacy, tolerability and safety of azithromycin and co-amoxiclav in the treatment of sinusitis in adults; Clement PA et al.; The efficacy, tolerability and safety of azithromycin and co-amoxiclav in the treatment of non-severe acute maxillary/ethmoidal sinusitis were compared in a randomized, open clinical trial in 254 adult patients . The predominant pathogens were Streptococcus pneumoniae and Haemophilus influenzae (83 patients) . Azithromycin was administered orally to 165 patients at a single daily dose of 500 mg for 3 days, and co-amoxiclav (4:1) to 89 patients, at a dose of 500 mg three times daily for 10 days . The overall clinical response rates were 87.5% for azithromycin and 83.7% for co-amoxiclav at follow-up (day 21-28) . Microbiological responses to both drugs were good, with only five patients in each group having a persistent infection after treatment . Both drugs were well tolerated and produced similar incidences of adverse events, which were mostly gastrointestinal . Azithromycin was as effective, and as well tolerated as co-amoxiclav, and its shorter simpler dosing regime may offer advantages in compliance and cost.

Infect Immun, 1998 Jun, 66(6), 2914 - 21
Haemophilus ducreyi infection causes basal keratinocyte cytotoxicity and elicits a unique cytokine induction pattern in an In vitro human skin model; Hobbs MM et al.; Haemophilus ducreyi is the etiologic agent of the sexually transmitted genital ulcer disease chancroid . Predominantly a cutaneous pathogen, H . ducreyi is present in chancroid ulcers that are characterized by extensive neutrophil accumulation in intraepidermal lesions accompanied by a mononuclear infiltrate in the dermis . We used an in vitro human skin model composed of foreskin fibroblasts and keratinocytes to examine host skin cell interactions with H . ducreyi 35000 . Bacteria replicated and persisted in artificial skin for at least 14 days . We observed H . ducreyi inside suprabasal keratinocytes using transmission electron microscopy . Although no bacteria were seen in the basal keratinocyte region, these cells were disrupted in infected cocultures . H . ducreyi infection stimulated increased secretion of interleukin-6 (IL-6) and IL-8 by skin cells . Conversely, tumor necrosis factor alpha and IL-1alpha levels were not elevated . IL-8 produced in response to H . ducreyi infection may be involved in recruiting polymorphonuclear leukocytes and other inflammatory cells, thereby contributing to the tissue necrosis and ulcer formation characteristic of chancroid.

J Biol Chem, 1998 May 22, 273(21), 12827 - 31
Molecular and functional properties of a calpain activator protein specific for mu-isoforms; Melloni E et al.; A natural calpain activator protein has been isolated from bovine brain and characterized in its properties and molecular structure . The protein is a homodimer with a molecular mass of about 30 kDa and results in being almost identical to UK114 goat liver protein . Significant similarities with mouse HR12 protein were also observed, whereas a lower degree of similarity was found with a family of heat-responsive proteins named YJGF and YABJ from Haemophilus influenzae and Bacillus subtilis, respectively . The brain activator expresses a strict specificity for the mu-calpain isoform, being completely ineffective on the m-calpain form . As expected, also UK114 was found to possess calpain-activating properties, indistinguishable from those of bovine brain activator . A protein showing the same calpain-activating activity has been also isolated from human red cells, indicating that this factor is widely expressed . All these activators are efficient on mu-calpain independently from the source of the proteinase . The high degree of specificity of the calpain activator for a single calpain isoform may be relevant for the understanding of sophisticated intracellular mechanisms underlying intracellular proteolysis . These data are indicating the existence of a new component of the Ca2+-dependent proteolytic system, constituted of members of a chaperonin-like protein family and capable of promoting intracellular calpain activation.

Aust N Z J Public Health, 1998 Feb, 22(1), 67 - 72
Measuring the impact of conjugate vaccines on invasive Haemophilus influenzae type b infection in Western Australia; Bower C et al.; Haemophilus influenzae type b (Hib) causes serious infections in 26-59 per 100,000 non-Aboriginal Australian children under five years of age . Aboriginal children suffer much higher rates of infection (> or = 150 per 100,000), and at an earlier age, and have a greater risk of death and disability due to Hib infection . In 1992 and 1993, four conjugate Hib vaccines were introduced in Australia, and a nationally funded program of infant vaccination was begun in July 1993 . This study aimed at evaluating the effectiveness of Hib vaccination in Aboriginal and non-Aboriginal children in Western Australia using a population-based active surveillance system for non-Aboriginal children and a case control study for Aboriginal children . The incidence of invasive Hib disease in non-Aboriginal children fell from 30.9 per 100,000 before vaccination was available to 6.3 per 100,000 in the second year after its introduction . The vaccine efficacy was estimated to be 80 per cent for Aboriginal children (odds ratio 0.20, 95 per cent CI 0.01-2.76) and, after adjustment for confounders, 75 per cent (odds ratio 0.25, CI 0.02-3.66) . Based on the adjusted value (75 per cent), and using a Bayesian approach, we estimate that the posterior probability was 0.55 that the true vaccine efficacy is greater than 70 per cent, and 0.69 that the efficacy is greater than 50 per cent . We conclude that Hib vaccination is effective in preventing invasive Hib disease in Aboriginal and non-Aboriginal children in Australia.

FEBS Lett, 1998 Apr 10, 426(1), 1 - 5
Differential genome analysis applied to the species-specific features of Helicobacter pylori; Huynen M et al.; We introduce a simple and rapid strategy to identify genes that are responsible for species-specific phenotypes . The genome of a species that has a specific phenotype is compared with at least one, closely related, species that lacks this phenotype . Homologous genes that are shared among the species compared are identified and discarded from the list of candidates for species-specific genes . The process is automated and rapidly yields a small subset of the genome that likely contains genes responsible for the species-specific features . Functions are assigned to the genes, and dubious annotations are filtered out . Information is extracted not only from the presence of genes, but also from their absence with respect to known phenotypes . We have applied the technique to identify a set of species-specific genes in Helicobacter pylori by comparing it with its closest relatives for which complete genome sequences are available, Haemophilus influenzae and Escherichia coli . Of the genes of this set for which functional features can be obtained, a large fraction (63%, 123 proteins) is (potentially) involved in H . pylori's interaction with its host . We hypothesize that a family of outer membrane proteins is critical for the ability of H . pylori to colonize host cells in highly acidic environments.

J Antimicrob Chemother, 1998 Apr, 41(4), 489 - 92
Bacterial interference in the nasopharynx following antimicrobial therapy of acute otitis media; Brook I et al.; The effect on the nasopharyngeal bacterial flora of therapy for 10 days with co-amoxiclav or cefprozil was studied in 50 children with acute otitis media . Before therapy, potential pathogens (Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis) were isolated in 14 (56%) of those treated with co-amoxiclav and 15 (60%) of those treated with cefprozil . Following therapy, the reduction in the number of these pathogens was the same in the two groups . However, differences between the groups were noted in the recovery of organisms with interfering capability, namely alpha-haemolytic streptococci, Peptostreptococcus anaerobius and Prevotella melaninogenica . Fifty interfering organisms were recovered from each group before therapy . After therapy with co-amoxiclav or cefprozil their number declined to 11 and 42, respectively (P< 0.001).

J Antimicrob Chemother, 1998 Apr, 41(4), 485 - 8
Serum bactericidal activity of newer oral cephalosporins in healthy volunteers; Dan M et al.; The serum bactericidal activity of three oral cephalosporins was studied in 12 volunteers, after administration of single doses of cefuroxime axetil 250 mg, cefixime 200 mg, cefixime 400 mg and cefetamet pivoxil 500 mg . Serum bactericidal activity against clinical isolates of Streptococcus pyogenes, Streptococcus pneumoniae, Haemophilus influenzae and Klebsiella pneumoniae was measured by a standardized microdilution method . Cefuroxime axetil demonstrated the best bactericidal activity against Gram-positive organisms and cefixime was the most bactericidal against Gram-negative bacteria.

Diagn Microbiol Infect Dis, 1998 May, 31(1), 313 - 25
In vitro evaluation of sparfloxacin activity and spectrum against 24,940 pathogens isolated in the United States and Canada, the final analysis; Jones RN et al.; Sparfloxacin, a recently marketed oral fluoroquinolone, was tested against 24,940 recent clinical strains isolated from blood stream and respiratory tract cultures at 187 hospitals in the USA and Canada . Sparfloxacin activity was compared with 5 to 13 antimicrobial agents using either Etest (AB BIODISK, Solna, Sweden) and a reference broth microdilution or a standardized disk diffusion method . When applying recommended MIC breakpoint criteria of sparfloxacin susceptibility (< or = 0.5 microgram/mL) for Streptococcus pneumoniae (4,410 strains) and other Streptococcus spp . (554 isolates), 93% and 88% were inhibited, respectively . Furthermore, at < or = 1 microgram/mL sparfloxacin susceptibility rates for streptococci increased to 98% overall and 99.3% for S . pneumoniae . In contrast, only 46% and 68% of pneumococci were susceptible to ciprofloxacin (MIC90, 3 micrograms/mL; susceptible at < or = 1 microgram/mL) and penicillin (MIC90, 1.5 microgram/mL; susceptible at < or = 0.06 microgram/mL), respectively . Differences between regions in the USA for rates of penicillin-resistant pneumococcal strains were observed (greatest resistances in southeast and midwest), but results indicate that the sparfloxacin potency was not adversely influenced (MIC90, 0.5 microgram/mL) . Also pneumococcal isolates from the lower respiratory tract were more resistant to penicillin and other beta-lactams . Nearly all Haemophilus species and Moraxella catarrhalis strains, including those harboring beta-lactamases, were susceptible to tested fluoroquinolones (sparfloxacin, ciprofloxacin), amoxicillin/clavulanic acid, and newer oral cephalosporins . Sparfloxacin was very active against oxacillin-susceptible Staphylococcus aureus (MIC90, 0.12 microgram/mL; 96-97% susceptible), Klebsiella spp . (MIC90 0.12 microgram/mL), and other tested enteric bacilli (92-95% susceptible) . Comparisons between the broth microdilution MIC and disk diffusion interpretive results demonstrated excellent intermethod susceptibility category agreement (> 95%) using current sparfloxacin breakpoints, but some compounds (cefpodoxime disk diffusion tests for S . aureus) may require modifications . These results demonstrate that new Gram-positive focused fluoroquinolones (sparfloxacin) possess an excellent in vitro activity and spectrum against pathogens that cause respiratory tract infections . This spectrum of activity includes strains resistant to other antimicrobial classes, including the oral cephalosporins, macrolides, amoxicillin/clavulanic acid, and earlier fluoroquinolones (ciprofloxacin, ofloxacin) . Overall, sparfloxacin inhibited 89% to nearly 100% of the isolates (species variable) tested against those species against which it has Food and Drug Administration indications for clinical use.

Clin Otolaryngol, 1998 Apr, 23(2), 181 - 5
Bacteriology of normal and diseased tonsils assessed by fine-needle aspiration: Haemophilus influenzae and the pathogenesis of recurrent acute tonsillitis; Gaffney RJ et al.; The pathogenesis of recurrent tonsillitis is largely unknown . Selection of appropriate antibiotic therapy for patients with recurrent tonsillitis is difficult because of the limitations of traditional methods of sampling tonsillar microflora and the increasing incidence of beta-lactamase producing bacteria in the tonsil . In addition, little attention has been paid to the bacteriology of normal tonsils . The tonsil core bacteria was assessed in 124 patients with recurrent acute tonsillitis . Fifty-five of these patients were randomly selected for fine-needle aspiration which revealed a similar profile of bacteria in 85% . Fine-needle aspiration of 10 normal tonsils found few pathogens; the predominant organisms being normal flora . No Haemophilus influenzae were detected in this control group . This study demonstrates the accuracy of fine-needle aspiration in identifying tonsil core bacteriology and its suitability in the clinical setting . It reports on the flora of normal healthy tonsils and it highlights the association between H . influenzae and recurrent acute tonsillitis.

Gene, 1998 Mar 27, 210(1), 117 - 25
Cloning and expression of Staphylococcus aureus and Treptococcus pyogenes murD genes encoding uridine diphosphate N-acetylmuramoyl-L-alanine:D-glutamate ligases; El-Sherbeini M et al.; Bacterial UDP-N-acetylmuramyl-L-alanine:D-glutamate ligase (MurD), a cytoplasmic peptidoglycan biosynthetic enzyme, catalyzes the ATP-dependent addition of D-glutamate to an alanyl residue of the UDP-N-acetylmuramyl-L-alanine precursor, generating the dipeptide . The murD gene was cloned from both Staphylococcus aureus and Streptococcus pyogenes . Sequence analysis of the S . aureus murD gene revealed an open reading frame of 449 amino acids . The deduced aa sequence of S . aureus MurD is highly homologous to MurD from Escherichia coli, Haemophilus influenzae, Bacillus subtilis and St . pyogenes . Recombinant MurD protein from both S . aureus and St . pyogenes was separately overproduced in E . coli and purified as His-tagged fusion . Both recombinant enzymes catalyzed the ATP-dependent addition of D-glutamate to the precursor sugar peptide.

Chest, 1998 May, 113(5), 1195 - 200
Comparative study of the clinical presentation of Legionella pneumonia and other community-acquired pneumonias; Sopena N et al.; The aim of this study was to compare the clinical, biological, and radiologic features of presentation in the emergency ward of community-acquired pneumonia (CAP) by Legionella pneumophila (LP) and other community-acquired bacterial pneumonias to help in early diagnosis of CAP by LP . Three hundred ninety-two patients with CAP were studied prospectively in the emergency department of a 600-bed university hospital . Univariate and multivariate analyses were performed to compare epidemiologic and demographic data and clinical, analytical, and radiologic features of presentation in 48 patients with CAP by LP and 125 patients with CAP by other bacterial etiology (68 by Streptococcus pneumoniae, 41 by Chlamydia pneumoniae, 5 by Mycoplasma pneumoniae, 4 by Coxiella burnetii, 3 by Pseudomonas aeruginosa, 2 by Haemophilus influenzae, and 2 by Nocardia species . Univariate analysis showed that CAP by LP was more frequent in middle-aged, male healthy (but alcohol drinking) patients than CAP by other etiology . Moreover, the lack of response to previous beta-lactamic drugs, headache, diarrhea, severe hyponatremia, and elevation in serum creatine kinase (CK) levels on presentation were more frequent in CAP by LP, while cough, expectoration, and thoracic pain were more frequent in CAP by other bacterial etiology . However, multivariate analysis only confirmed these differences with respect to lack of underlying disease, diarrhea, and elevation in the CK level . We conclude that detailed analysis of features of presentation of CAP allows suspicion of Legionnaire's disease in the emergency department . The initiation of antibiotic treatment, including a macrolide, and the performance of rapid diagnostic techniques are mandatory in these cases.

J Comp Pathol, 1998 Apr, 118(3), 231 - 43
Ultrastructural study of porcine alveolar macrophages infected in vitro with porcine reproductive and respiratory syndrome (PRRS) virus, with and without Haemophilus parasuis; Segales J et al.; Two experiments were designed to study ultrastructural changes in porcine alveolar macrophages (PAM) inoculated with porcine reproductive and respiratory syndrome (PRRS) virus (experiment 1) and with PRRS virus and Haemophilus parasuis (experiment 2) . In both experiments, the viral infectious dose represented a "multiplicity of infection" of 1 . Viral infection alone induced minimal ultrastructural changes at this dose, consisting only of an increase in lysosome numbers . Mixed viral and bacterial infection induced the production of greatly increased numbers of phagosomes and phagolysosomes . The PAM were of low efficacy in phagocytizing H . parasuis . PRRS virus infection had only a minimal effect on the phagocytosis of H . parasuis by PAM . It is suggested that the virus induces PAM activation rather than PAM destruction.

Trop Med Int Health, 1998 Mar, 3(3), 205 - 9
Bacteria and viruses that cause respiratory tract infections during the pilgrimage (Haj) season in Makkah, Saudi Arabia; El-Sheikh SM et al.; OBJECTIVE: To determine the incidence and type of RTI-causing bacteria and viruses during a period of epidemic infections . METHOD: A total of 395 sputum specimens and 761 throat swabs were collected during the 1991 and 1992 pilgrimage seasons (Haj to Makkah Al-Mukarama, Saudi Arabia) from patients referred to one hospital and three dispensaries with symptoms of respiratory tract infections . All 761 throat swabs of both Haj seasons were also screened for the presence of viral pathogens with monoclonal antibodies specific for 7 viruses known to cause respiratory infections . RESULTS: Bacterial pathogens were detected in 118 (29.9%) specimens . During the 1991 Haj season Haemophilus influenzae was the most frequent bacterial pathogen detected (10%), followed by Klebsiella pneumoniae (5.2%), Streptococcus pneumoniae (4.8%), Staphylococcus aureus (3.8%) and Streptococcus pyogenes (2.4%) . In the 1992 Haj season Klebsiella pneumoniae was predominant (15.1%), followed by Haemophilus influenzae and Streptococcus pneumoniae (12.3%) . Screening of all sputum specimens for acid-fast bacteria showed that the overall incidence rate of tuberculosis was 1% . Cultures from the 761 throat swabs were largely negative for bacteria except for Streptococcus pyogenes isolated from 7 patients . Viruses were detected in 148 (19.5%) specimens with influenza A and adenovirus being the most common viruses . CONCLUSION: The pattern of virus prevalence in the 1991 and 1992 pilgrimage seasons was identical: influenza A and adenovirus predominated . Thus these two viruses should be targeted in future prophylactic measures.

Pol Merkuriusz Lek, 1998 Feb, 4(20), 78 - 80
{Resistance of Haemophilus sp . to antibiotics}; Janicka G et al.; The present study was undertaken to determine the in vitro drug resistance of Haemophilus influenzae (68 isolates) and H . parainfluenzae (17 isolates) . The tests susceptibility to Ampicillin, Amoxicilin/Clavulanic Acid, Cefaclor, Cefuroxime, Cotrimoxazole, Aztreonam, Ceftriaxone, Tetracycline, Ciprofloxacin, Rifampicin and Chloramphenicol were performed with a standard disk-diffusion method . The NCCLS methodology and susceptibility interpretative criteria were applied as described by the disk manufacturer . Beta-lactamase production was detected with nitrocefin impregnated disk (Cefinase, BBL Microbiology System) . Resistance in nosocomially acquired Haemophilus isolates to several antibiotics was observed . Of the Haemophilus isolates 28.2% were Ampicillin in resistant, all were susceptible to the combination of Amoxicillin/Clavulanic acid . The Ampicillin-resistant strains were beta-lactamase producers . We observed the high resistance (70.1%) to Tetracycline and (28.2%) to SXT (Cotrimoxazole) . All isolates of Haemophilus were susceptible to Ciprofloxacin . The low resistance percentages to Rifampin (1.2%), Aztreonam (3.5%) and Chloramphenicol (3.5%) was observed.

J Paediatr Child Health, 1998 Apr, 34(2), 175 - 8
Documentation of children's vaccination status in child care centres in Victoria; Thompson SC et al.; OBJECTIVE: To assess the record-keeping of child care centres in Victoria with respect to children's vaccination status . METHODOLOGY: A random sample of 113 centres from a list of over 800 registered Victorian child care centres received a mailed questionnaire on characteristics and policies of the centre, including documentation of attending children's vaccination status . RESULTS: The response rate was 86.7%; more than 95% of centres had children under two in care . Only 85% of centres kept any record of children's immunisation status, with smaller centres and class 2 centres (occasional care centres) significantly less likely to keep records . Records were updated irregularly . Fewer than half of the centres studied kept a record of whether children had been vaccinated against Haemophilus influenzae type b . CONCLUSIONS: Despite the importance of documenting children's vaccination status, many child care centres have failed to develop adequate systems to record or update records of children's immunisation . This makes exclusion during outbreaks of a vaccine-preventable disease difficult . The ongoing process of accreditation may eventually improve performance, but both legislation and a better educational strategy to improve practice in this area are needed.

J Paediatr Child Health, 1998 Apr, 34(2), 109 - 11
Geographic differences in bacterial meningitis: less may be as interesting as more; McIntyre P; Many reports in the last decade have described populations with a high incidence of bacterial meningitis, especially amongst indigenous groups in industrialised countries, such as North American Eskimos and Apache Indians and Australian Aborigines, particularly with meningitis due to Haemophilus influenzae type b (Hib) . Lack of evidence that invasive Hib disease, including meningitis, is a significant health problem has been attributed to lack of appropriate data, either due to lack of laboratory and clinical facilities, such as in most less industrialised countries, or lack of study . Host differences in immune response, though known to be important for individual susceptibility to Hib disease and bacterial meningitis, have not been thought important on a population level . Good quality epidemiologic data now available from Hong Kong and Japan, based on sound laboratory methods, have shown bacterial meningitis, particularly due to Hib and Neisseria meningitidis, to be significantly less common than in predominantly Caucasian populations in various industrialised countries . Differences in host immune response to these capsular polysaccharides seems the most likely explanation for this observation . It is interesting that other immunologically mediated disorders such as Kawasaki disease and systemic lupus erythematosis have a relatively high incidence in Sino Japanese populations, lending plausibility to inherited differences in immune response as a mechanism for these observations.

Sex Transm Dis, 1998 May, 25(5), 237 - 42
A serosurvey of Haemophilus ducreyi, syphilis, and herpes simplex virus type 2 and their association with human immunodeficiency virus among female sex workers in Lagos, Nigeria; Dada AJ et al.; BACKGROUND AND OBJECTIVES: To determine the prevalence rates of serological reactivity of Haemophilus (H.) ducreyi, Treponema pallidum, and herpes simplex virus type 2 (HSV-2) antibodies among female sex workers (FSWs) and their association with human immunodeficiency virus (HIV) antibody status . STUDY DESIGN: Cross-sectional, standard serological assays were used for syphilis, HSV-2 and HIV; a modified enzyme-linked immunosorbent assay (ELISA) was used to detect specific anti-H . ducreyi immunoglobulin (Ig) G and IgA antibodies . RESULTS: Seroprevalence rates were 86% for anti-H . ducreyi IgG and 69% for anti-H . ducreyi IgA; 4% for rapid plasma reagin (RPR) and Treponema palladium hemagglutination assay (TPHA) confirmed syphilis; 59% for HSV-2; 12% for HIV-1 and 2% for HIV-2 . Lower-class FSWs were significantly more likely than upper-class FSWs to be H . ducreyi seropositive (IgG: OR = 42.7; IgA: OR = 7.6) and have current or past syphilis infection (RPR: OR = 3.5; RPR and TPHA: OR = 4.5) . The presence of syphilis increased significantly with older age (P-trend < 0.001) . Non-Nigerian FSWs had significantly higher reactivity to chancroid (IgG: OR = 3.5; IgA: OR = 1.8) and borderline reactivity to syphilis (RPR: OR = 1.6; TPHA: OR = 2.0) . A history of sex with non-Nigerian Africans was significantly associated with chancroid reactivity and borderline significant with syphilis serostatus . H . ducreyi seropositivity was significantly more likely in FSWs with HSV-2 (OR = 2.4) and syphilis (OR = 5.6) . Chancroid and HSV-2 antibodies were also more common in HIV-infected FSWs . CONCLUSION: The prevalence of H . ducreyi antibodies is the highest rate that has been reported . Our findings underscore the importance of an effective program to control GUDs as part of the strategy to prevent the potentially explosive spread of HIV in NigeriaPIP: Cross-sectional standard serologic assays were used to determine the prevalence of Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus type 2 (HSV-2) antibodies among 796 female commercial sex workers from Lagos, Nigeria, and their association with HIV antibody status . The seroprevalence rates were 86% for anti-H . ducreyi IgG and 69% for anti-H . ducreyi IgA, 4% for rapid plasma reagin and Treponema palladium hemagglutination assay confirmed syphilis, 59% for HSV-2, 12% for HIV-1, and 2% for HIV-2 . Lower-class sex workers were significantly more likely than upper-class sex workers to be H . ducreyi-positive and to have current or past syphilis infection . The presence of syphilis increased significantly with older age . Non-Nigerian sex workers had significantly higher reactivity to chancroid and borderline reactivity to syphilis . A history of sex with non-Nigerian Africans was significantly associated with chancroid reactivity and borderline significant with syphilis serostatus . H . ducreyi seropositivity was significantly more likely in female sex workers with HSV-2 and syphilis . Chancroid and HSV-2 antibodies were also more common in HIV-infected sex workers . The high prevalence of H . ducreyi antibodies detected in this study underscores the importance of an effective program to control genital ulcerative disease as part of the strategy to prevent the spread of HIV in Nigeria .

Acta Otolaryngol, 1998 Mar, 118(2), 211 - 5
Changes in mucosal goblet cell density in acute otitis media caused by non-typeable Haemophilus influenzae; Caye-Thomasen P et al.; The correlation between secretory otitis media and increased goblet cell density in the middle ear mucosa is well established . Previous studies have shown that a single episode of acute otitis media caused by Streptococcus pneumoniae is followed by increased goblet cell density for a period of at least 6 months, conceivably predisposing a subsequent development of secretory otitis media . In this study, 25 rat middle ears were inoculated with non-typeable Haemophilus influenzae in order to determine the effect on mucosal goblet cell density . Five rats were killed on days 4, 8, 16, 60 and 180 postinoculation, followed by dissection, staining and whole-mount embedding of the middle ear mucosae . The goblet cell density was determined in 24 well-defined localities . Compared with 25 normal middle ears, the goblet cell density was significantly increased in almost all localities, at all days on which the animals were killed . Thus, increased goblet cell density and enlargement of mucosal areas containing goblet cells persisted 6 months after the acute incident . The induced increase of goblet cell density was higher than the increase following inoculation of S . pneumoniae . We conclude that acute otitis media caused by non-typeable H . influenzae is followed by a longstanding increase in mucosal secretory capacity, likely to predispose a subsequent development of secretory otitis media.

Acta Otolaryngol, 1998 Mar, 118(2), 206 - 10
Aerobic and anaerobic bacteriology of otorrhea associated with tympanostomy tubes in children; Brook I et al.; The microbiology of in 55 ear aspirates obtained from 34 children with chronic otorrhea was studied . Aspiration of the middle ear exudate was done immediately following removal of tympanostomy tube (TT) . The middle ear aspirates and swab specimens of the external auditory canals were cultured for aerobic and anaerobic bacteria . Sixty-five isolates were recovered only from the middle ears, 73 only from the external ear canals, and 73 were present at both sites . Analysis of the 138 middle ear isolates demonstrated the recovery of aerobic bacteria only in 28 patients (50%), anaerobes only in seven (13%), and both aerobes and anaerobes in 20 (36%) . There were 77 aerobic and 61 anaerobic isolates . Commonly recovered aerobes were Pseudomonas aeruginosa (17 isolates), Staphylococcus aureus (11), Proteus sp . (7), Moraxella catarrhalis (6), Klebsiella pneumoniae (5) and non-typable Haemophilus influenzae (5) . Commonly isolated anaerobes were Peptostreptococcus sp . (25 isolates), Prevotella sp . (10), Bacteroides sp . (8) and Fusobacterium sp . (6) . Pseudomonas aeruginosa and S . aureus were more often isolated in children older then 6 years . These findings demonstrate the polymicrobial bacteriology of TT-related otorrhea in children . Specimens collected from the external auditory canals can be misleading . Reliable information can be obtained from the ear exudes when collected through the TT or through the open perforation after their removal.

Ann Allergy Asthma Immunol, 1998 Apr, 80(4), 357 - 62
Open-label assessment of levofloxacin for the treatment of acute bacterial sinusitis in adults; Sydnor TA et al.; PURPOSE: To evaluate the efficacy and safety of levofloxacin (500 mg orally once daily for 10 to 14 days) in treating adult outpatients with acute bacterial sinusitis . PATIENTS AND METHODS: A total of 329 patients enrolled in the study at 24 centers . All patients had a pre-therapy Gram's stain and culture of sinus exudate obtained by antral puncture or nasal endoscopy . Clinical response was assessed on the basis of signs and symptoms and sinus radiograph or computed tomography results . Microbiologic cure rates were determined on the basis of presumed plus documented eradication of the pre-therapy pathogen(s) . RESULTS: The most common pathogens were Haemophilus influenzae, Streptococcus pneumoniae, Staphylococcus aureus, and Moraxella catarrhalis . Of 300 clinically evaluable patients, 175 (58%) were cured and 90 (30%) were improved at the post-therapy evaluation, resulting in a clinical success rate of 88% . Thirty-five patients (12%) clinically failed treatment . The microbiologic eradication rate (presumed plus documented) among 138 microbiologically evaluable patients was 92% . Microbiologic eradication rates (presumed plus documented) of the most common pathogens ranged from 93% (M . catarrhalis) to 100% (S . pneumoniae) at the post-therapy visit . All but one of the 265 patients who were cured or improved at post-therapy returned for a long-term follow-up visit; 243 (92%) remained well 4 to 6 weeks after therapy; and 21 (8%) had a relapse of symptoms . Adverse events considered to be related to levofloxacin administration were reported by 29 patients (9%) . The most common drug-related adverse events were diarrhea, flatulence, and nausea; most adverse events were mild to moderate in severity . CONCLUSION: The results of this study indicate that levofloxacin 500 mg once daily is an effective and safe treatment for acute bacterial sinusitis.

Eur Respir J, 1998 Jan, 11(1), 86 - 90
Effect of salmeterol on Haemophilus influenzae infection of respiratory mucosa in vitro; Dowling RB et al.; Haemophilus influenzae is a common bacterial pathogen causing human respiratory tract infections . We have previously shown that the beta2-agonist salmeterol reduces damage to the respiratory mucosa caused by Pseudomonas aeruginosa in vitro . We have now investigated the effect of salmeterol on H . influenzae infection of adenoid tissue in an organ culture by scanning electron microscopy . Tissue was preincubated with or without salmeterol (4x10(-7)M), prior to infection with H . influenzae and incubated for 12 or 24 h . Infected organ cultures had increased epithelial damage and decreased numbers of both ciliated and unciliated cells at 12h, which were significantly different (p < or = 0.01) from the controls at 24 h . Salmeterol (4x10(-7)M) significantly (p < or = 0.03) reduced damage and loss of ciliated cells in infected organ cultures at both 12 and 24, and significantly (p < or = 0.03) reduced loss of unciliated cells at 24 h . Salmeterol had no effect on the density of bacteria adhering to each individual mucosal feature or the total number of bacteria adhering to the organ culture . These results suggest that salmeterol protects the respiratory epithelium against Haemophilus influenzae-induced damage . The mechanism of salmeterol cytoprotection and its potential clinical relevance remain to be investigated.

Protein Expr Purif, 1998 Apr, 12(3), 295 - 304
Expression in Escherichia coli of the putative N-acetylneuraminate lyase gene (nanA) from Haemophilus influenzae: overproduction, purification, and crystallization; Lilley GG et al.; The cloning and expression of the Haemophilus influenzae gene, nanA, for the putative N-acetylneuraminate lyase enzyme, also known as N-acetylneuraminic acid aldolase or sialic acid aldolase, are reported . The gene was isolated from ATCC type strain 49247 and cloned into the Escherichia coli expression vector pKKtac, which contained the strong tac promoter . Gene expression was compared with the homologous E . coli npl gene coding for the lyase . Purification protocols for the products of the nanA and npl genes are presented . Activity analysis showed that the nanA gene product is a sialic acid aldolase with more than threefold greater specific activity (6.9 IU/mg) than the enzyme from E . coli (</=2 IU/mg) . A method for the provision of lyase orthorhombic crystals is reported . These crystals diffract to better than 2.0 A, which paves the way to the solution of the enzyme's three-dimensional structure .

Genome Res, 1998 Mar, 8(3), 203 - 10
Reconstruction of amino acid biosynthesis pathways from the complete genome sequence; Bono H et al.; The complete genome sequence of an organism contains information that has not been fully utilized in the current prediction methods of gene functions, which are based on piece-by-piece similarity searches of individual genes . We present here a method that utilizes a higher level information of molecular pathways to reconstruct a complete functional unit from a set of genes . Specifically, a genome-by-genome comparison is first made for identifying enzyme genes and assigning EC numbers, which is followed by the reconstruction of selected portions of the metabolic pathways by use of the reference biochemical knowledge . The completeness of the reconstructed pathway is an indicator of the correctness of the initial gene function assignment . This feature has become possible because of our efforts to computerize the current knowledge of metabolic pathways under the KEGG project . We found that the biosynthesis pathways of all 20 amino acids were completely reconstructed in Escherichia coli, Haemophilus influenzae, and Bacillus subtilis, and probably in Synechocystis and Saccharomyces cerevisiae as well, although it was necessary to assume wider substrate specificity for aspartate aminotransferases.

Mol Biol Evol, 1998 May, 15(5), 583 - 9
The frequency distribution of gene family sizes in complete genomes; Huynen MA et al.; We compare the frequency distribution of gene family sizes in the complete genomes of six bacteria (Escherichia coli, Haemophilus influenzae, Helicobacter pylori, Mycoplasma genitalium, Mycoplasma pneumoniae, and Synechocystis sp . PCC6803), two Archaea (Methanococcus jannaschii and Methanobacterium thermoautotrophicum), one eukaryote (Saccharomyces cerevisiae), the vaccinia virus, and the bacteriophage T4 . The sizes of the gene families versus their frequencies show power-law distributions that tend to become flatter (have a larger exponent) as the number of genes in the genome increases . Power-law distributions generally occur as the limit distribution of a multiplicative stochastic process with a boundary constraint . We discuss various models that can account for a multiplicative process determining the sizes of gene families in the genome . In particular, we argue that, in order to explain the observed distributions, gene families have to behave in a coherent fashion within the genome; i.e., the probabilities of duplications of genes within a gene family are not independent of each other . Likewise, the probabilities of deletions of genes within a gene family are not independent of each other.

J Antimicrob Chemother, 1998 Mar, 41 Suppl B, 75 - 80
Comparison of roxithromycin with cefixime in the treatment of adults with community-acquired pneumonia; Salvarezza CR et al.; This study compared the efficacy and tolerability of once-daily dosing with either roxithromycin or cefixime in previously healthy adult patients aged between 18 and 60 with markers of uncomplicated community-acquired pneumonia (CAP) in three outpatient clinics in an open, randomized study . Sixty patients were enrolled: 17 males and 13 females received roxithromycin 300 mg once daily for 8-10 days and 22 males and eight females received 400 mg cefixime once daily for the same period . All patients were assessed clinically, radiologically and bacteriologically before inclusion, immediately after the study and approximately 1 month later . The most common pathogen isolated from sputum was Streptococcus pneumoniae (in 26 (43%) of 60 patients), with mixed organisms isolated from the sputum of 18 (30%) of 60 patients . Staphylococcus aureus, Haemophilus influenzae or Moraxella catarrhalis occurred in 11/60 patients, and atypical pathogens were detected by serology in 7/26 cases in the roxithromycin group and 3/23 in the cefixime group . The severity of infection was rated as mild to moderate at the beginning of the trial . At the end of the study treatment period, clinical cure rates were 30/30 (100%) for roxithromycin and 28/30 (94%) for cefixime, with one patient on cefixime being classed as a partial responder and one patient being classed as a failure and withdrawn . However, radiological abnormalities persisted in three patients on roxithromycin and one on cefixime . Of the 59 patients who completed the study, none required further antibiotic therapy . No abnormal laboratory parameters or adverse events were reported in either group . Roxithromycin at a daily dose of 300 mg was an effective and well-tolerated treatment for the empirical treatment of mild to moderate CAP in this group of patients.

Microbiology, 1998 Apr, 144 ( Pt 4), 1013 - 9
The rrs (16S)-rrl (23S) ribosomal intergenic spacer region as a target for the detection of Haemophilus ducreyi by a heminested-PCR assay; Gu XX et al.; The intergenic spacer region between the rrs and rrl ribosomal RNA genes of Haemophilus ducreyi was analysed and the DNA sequence was used for the selection of specific PCR primers . A highly sensitive and specific heminested-PCR assay for the identification of H . ducreyi was developed . The assay showed a sensitivity of 96% on genital ulcer specimens from patients with clinically diagnosed chancroid, compared with a sensitivity of 56% for culture methods . These results indicate that this PCR assay has the potential to become an accurate and easy reference method for the detection of H . ducreyi.

Microbiology, 1998 Apr, 144 ( Pt 4), 975 - 83
Comparative analysis of Pseudomonas aeruginosa penicillin-binding protein 7 in the context of its membership in the family of low-molecular-mass PBPs; Song J et al.; The Pseudomonas aeruginosa pbpG gene encoding penicillin-binding protein 7, a homologue of the Escherichia coli gene encoding a DD-endopeptidase, was cloned and sequenced, pbpG was located immediately downstream of the phenylalanine hydroxylase (phh) operon . DNA sequencing revealed an open reading frame of 936 bp (starting with a GTG codon) which encodes a protein of 34,115 Da . N-terminal amino acid sequencing confirmed the presence of a cleavable N-terminal signal peptide of 23 amino acids . Verification that the protein is a penicillin-binding protein was directly demonstrated by labelling with 125I-labelled penicillin X . Inactivation of P . aeruginosa pbpG by interposon mutagenesis resulted in no obvious phenotypic changes, but when P . aeruginosa PbpG was overexpressed in E . coli using a T7 expression system, cell lysis resulted . P . aeruginosa PbpG resembled E . coli PbpG in being associated with the membrane fraction . Two additional members of the PbpG subfamily were identified in the database . P . aeruginosa PbpG shows 63% identity with E . coli penicillin-binding protein 7 (PbpG) and 60% identity with Vibrio cholerae PbpG, but only 23% identity with Haemophilus influenzae PbpG . The PbpG subfamily and three other subfamilies constituting the low-molecular-mass PBP protein family were analysed by multiple alignment of 26 sequences . PbpG exhibited the consensus motifs of other penicillin-binding proteins . Ten anchor residues were identified that are conserved at the family level within the superfamily of serine-active-site penicillin-interacting proteins.

Neurologia, 1998 Feb, 13(2), 92 - 3
{Haemophilus influenzae type B meningitis: typical and atypical presentation}; Sanchez JM et al.; We present 2 cases of Haemophilus influenzae meningitis . The first is a patient with atypical simptomatology: abdominal pain, fever and two days later pain in the back of his legs . Abdominal pathology was not found . The cerebrospinal fluid (CSF) showed polymorphonuclear cells, hyperproteinorachia and lowered glucose . CSF culture revealed Haemophilus influenzae, blood culture was sterile . The second had suffered surgery at maxilar and ethmoid sinuses four years before, and unknown germ meningitis 6 months before . Haemophilus influenzae was isolated from CSF cultures and CSF rhinorrhea was detected by isotopic cisternography.

J Antimicrob Chemother, 1998 Mar, 41(3), 411 - 5
Antimicrobial susceptibility of community-acquired lower respiratory tract bacterial pathogens isolated in the UK during the 1995-1996 cold season; Felmingham D et al.; The antimicrobial susceptibility of 1078 isolates of Haemophilus influenzae, 348 Streptococcus pneumoniae and 258 Moraxella catarrhalis was determined . Overall 15.1% of H . influenzae produced beta-lactamase; 98.8% were susceptible to co-amoxiclav, 85.8% to cefaclor, 96% to clarithromycin and 100% to ciprofloxacin . The majority (94.2%) of M . catarrhalis produced beta-lactamase . The overall prevalence of low-level penicillin resistance (MIC = 0.12-1 mg/L) amongst isolates of S . pneumoniae was 3.4% and that of high-level resistance (MIC > or = 2 mg/L) was 3.7% . Most (96.3%) of the isolates of S . pneumoniae were susceptible to amoxycillin (MIC < or = 0.5 mg/L), 96% to cefaclor (MIC < or = 8 mg/L), 90.7% to clarithromycin (MIC < or = 0.25 mg/L) and 89% to ciprofloxacin (MIC < or = 1 mg/L).

Med J Aust, 1998 Apr 6, 168(7), 344 - 8
Should third-generation cephalosporins be the empirical treatment of choice for severe community-acquired pneumonia in adults?
Paterson DL, Playford EG.
The choice of empirical treatment for community-acquired pneumonia (CAP) is highly controversial . Our survey of 42 Australian emergency department doctors showed that monotherapy with a third-generation cephalosporin was the preferred regimen for severe CAP (14/42; 33%) . We argue that cheaper regimens with a narrower spectrum are likely to be just as effective as third-generation cephalosporins and will have fewer adverse effects on the microbial ecology of hospitals . We suggest penicillin or ampicillin (to cover pneumococci--even if penicillin "resistant"--and Haemophilus influenzae), plus a macrolide (e.g., azithromycin or erythromycin; to cover Legionella and other "atypical" pathogens), plus a single large dose of an aminoglycoside (e.g., gentamicin; to cover gram-negative bacilli such as Klebsiella pneumoniae) as empirical therapy for severe CAP.

Biochem J, 1998 May 15, 332 ( Pt 1), 223 - 30
Enzyme-mediated cytosine deamination by the bacterial methyltransferase M.MspI; Zingg JM et al.; Most prokaryotic (cytosine-5)-DNA methyltransferases increase the frequency of deamination at the cytosine targeted for methylation in vitro in the absence of the cofactor S-adenosylmethionine (AdoMet) or the reaction product S-adenosylhomocysteine (AdoHcy) . We show here that, under the same in vitro conditions, the prokaryotic methyltransferase, M.MspI (from Moraxella sp.), causes very few cytosine deaminations, suggesting a mechanism in which M.MspI may avoid enzyme-mediated cytosine deamination . Two analogues of AdoMet, sinefungin and 5'-amino-5'-deoxyadenosine, greatly increased the frequency of cytosine deamination mediated by M.MspI presumably by introducing a proton-donating amino group into the catalytic centre, thus facilitating the formation of an unstable enzyme-dihydrocytosine intermediate and hydrolytic deamination . Interestingly, two naturally occurring analogues, adenosine and 5'-methylthio-5'-deoxyadenosine, which do not contain a proton-donating amino group, also weakly increased the deamination frequency by M.MspI, even in the presence of AdoMet or AdoHcy . These analogues may trigger a conformational change in the enzyme without completely inhibiting the access of solvent water to the catalytic centre, thus allowing hydrolytic deamination of the enzyme-dihydrocytosine intermediate . Under normal physiological conditions the enzymes M.HpaII (from Haemophilus parainfluenzae), M . HhaI (from Haemophilus hemolytica) and M.MspI all increased the in vivo deamination frequency at the target cytosines with comparable efficiency.

Pediatr Infect Dis J, 1998 Apr, 17(4), 309 - 12
A polymerase chain reaction for the diagnosis of Haemophilus influenzae type b disease in children and its evaluation during a vaccine trial; Hassan-King M et al.; BACKGROUND: Determination of the etiology of pneumonia in young children is difficult because blood culture, the usual method of diagnosis, is positive in only a small proportion of cases . For this reason vaccine trials that include bacterial pneumonia as an endpoint must be large . OBJECTIVES: To determine whether a diagnostic test based on a polymerase chain reaction could be used as an alternative to conventional blood culture for diagnosis of invasive Haemophilus influenzae type b (Hib) infections in young children investigated during the course of a large vaccine trial . METHODS: DNA was extracted from blood culture supernatants and probed for the presence of Hib DNA with a PCR assay with primers derived from the cap gene locus of Hib . Results of the PCR assay were compared with those obtained by conventional culture techniques . RESULTS: Blood cultures were obtained from 1544 children with suspected pneumonia, meningitis or septicemia and from 31 healthy control children who were contacts of cases . Blood culture supernatants were tested for Hib DNA in the PCR test . The sensitivity and specificity of a positive PCR test in blood culture supernatant as against culture of Hib from any normally sterile site were 100 and 99%, respectively . Eleven children had positive Hib PCR tests on blood culture supernatants but were negative by culture . In one of these cases Hib was isolated from a lung aspirate and in two other patients H . influenzae strains other than Hib were obtained from the cerebrospinal fluid . Eight of these 11 children were in the control group . When the results of the PCR assay were used to determine vaccine efficacy, a value of 86% was obtained compared with a figure of 95% obtained when conventional culture techniques were used . CONCLUSIONS: An Hib PCR assay on blood culture supernatants proved to be sensitive and specific for the diagnosis of Hib disease in children . The distribution of PCR-positive, culture-negative cases between Hib-vaccinated and control groups paralleled that of culture-positive cases, suggesting that most of these children had been infected with Hib . A trial of a highly efficacious vaccine provides a novel way for evaluating new diagnostic tests for which there is no standard diagnostic test of 100% reliability.

Pediatr Infect Dis J, 1998 Apr, 17(4), 294 - 304
Clinical acceptability and immunogenicity of a pentavalent parenteral combination vaccine containing diphtheria, tetanus, acellular pertussis, inactivated poliomyelitis and Haemophilus influenzae type b conjugate antigens in two-, four- and six-month-old Chilean infants; Lagos R et al.; BACKGROUND: In recent years additional parenteral vaccines have been recommended for routine immunization of infants in the US and elsewhere . The ability to administer multiple vaccines as a single injection without unacceptably increasing reactogenicity or decreasing immunogenicity of any component would offer many practical advantages . METHODS: A randomized, open, controlled trial was conducted to assess the tolerance profile and immunogenicity, as well as to identify potential antigenic interferences, resulting from administration of a parenteral combination vaccine for infants . The vaccine contains diphtheria and tetanus toxoids, acellular pertussis antigens (DTaP), enhanced inactivated poliovirus (eIPV) and Haemophilus influenzae type b-tetanus toxoid conjugate (PRP-T) . Infants (n=711) were randomly assigned to receive 1 of 5 regimens as the primary series at 2, 4 and 6 months of age, by group: (1) DTaP plus oral polio vaccine (OPV); (2) DTaP plus eIPV (separate injections); (3) DTaP-eIPV combined as a single injection; (4) DTaP-eIPV combined, plus a separate injection of PRP-T; or (5) DTaP-eIPV combined and reconstituting PRP-T, as a single injection . At 3, 5 and 7 months Groups 1, 2 and 3 received PRP-T . At 12 months all infants received a booster dose of DTaP reconstituting PRP-T as a single injection, plus a separate injection of measles, mumps and rubella vaccine . Groups 2, 3, 4 and 5 received OPV at 7 months, and all infants received OPV at 13 months . Serum immune responses were measured to the primary series at 2 and 7 months and to the booster dose at 12 and 13 months . RESULTS: Reaction rates were similar among groups . In the primary series combining eIPV with DTaP decreased geometric mean titers (GMTs) to diphtheria, tetanus and pertussis . In addition concomitant PRP-T (either simultaneous or combined) with DTaP-eIPV lowered anti-PRP and further decreased tetanus GMTs . Nonetheless in 100% of infants protective titers were achieved against diphtheria and tetanus (>0.01 IU/ml each) and against the poliovirus types 1, 2 and 3 after eIPV (Groups 2 to 5); 99% of infants (Groups 4 and 5) had protective titers against PRP (> or = 0.15 microg/ml) . After boosting with DTaP/PRP-T decreased GMTs to diphtheria and PRP antigens were observed in the groups that received DTaP and eIPV combined . Nonetheless protective titers to diphtheria, tetanus and PRP occurred consistently . In contrast concomitant PRP-T with DTaP-eIPV enhanced the pertussis GMTs . CONCLUSIONS: We conclude that combined DTaP, eIPV and PRP-T in a single injection is well-tolerated and elicits an acceptable immune response to each component.

Pediatr Infect Dis J, 1998 Apr, 17(4), 271 - 7; discussion 277-9
A survey about management of febrile children without source by primary care physicians; Wittler RR et al.; BACKGROUND: The management of young children with fever without source is controversial, and differences between physician specialties have been noted previously . The emergence of penicillin-resistant Streptococcus pneumoniae, the sharp decline in invasive Haemophilus influenzae infections in immunized populations and publication of practice guidelines have potentially altered physician practices . OBJECTIVE: To determine the present practice preferences of pediatricians, family medicine physicians (FP) and emergency medicine physicians (EP) . METHODS: We mailed a checklist survey to 1600 randomly selected pediatricians, family medicine practitioners (FP) and emergency medicine physicians (EP) in the United States and replicated the methodology of a 1991/1992 survey . Physicians were asked about their evaluation and management of children of various ages (3 weeks, 7 weeks, 4 months and 16 months) with fever without source . RESULTS: Most primary care physicians would admit the 3- and 7-week-old infants . For the 4-month-old infant 59% of EP, 45% of pediatricians and 28% of FP would give empiric antibiotic(s) as an outpatient (P=0.005 for FP compared with pediatricians and P=0.02 for EP compared with pediatricians) . The majority of physicians would manage the 16-month-old child as an outpatient without antibiotic therapy . Ceftriaxone was the preferred antibiotic for outpatient empiric therapy . There was a 3-fold increase (28% vs . 9%) for pediatricians in the use of empiric outpatient antibiotics for the 7-week-old infant in the present survey compared with the 1991/1992 survey . CONCLUSIONS: Physicians in the United States generally agree in their management of the young febrile infant, but with increasing patient age there is considerable variation . FP were the least aggressive in their evaluation and EP were the most aggressive.

J Clin Microbiol, 1998 May, 36(5), 1185 - 8
Two-step PCR-based assay for identification of bacterial etiology of otitis media with effusion in infected Lebanese children; Matar GM et al.; We developed and evaluated a two-step PCR-based assay with universal primers and genus- or species-specific primers for the detection of the most prevalent bacterial etiologies of otitis media with effusion (OME) in children from Lebanese hospitals . These etiologies included Haemophilus, Streptococcus, and Moraxella (Branhamella) catarrhalis, which were detected in middle-ear effusion (MEE) samples taken from children with OME . A total of 47 MEE samples were aspirated from 36 patients during insertion of a tympanostomy tube performed particularly for OME . The duration of effusion in all patients was > or =2 months . DNA was extracted from MEE samples, and PCR was initially done with DNA extracts by using the universal primers RW01 and DG74, which flank an approximately 370-bp fragment found in the 16S rRNA gene of all bacterial species . For the identification of specific bacteria, we used in three separate reaction mixtures the following genus- or species-specific primers: (i) a Haemophilus-specific probe (probe RDR125) as a primer along with DG74, (ii) a Streptococcus-specific primer (primer STR1; designed by us) along with DG74, and (iii) an M . catarrhalis-specific primer pair (primer pair MCA1-MCA2) . Thirty-five MEE samples (74.5%) gave the expected 370-bp band, indicating the presence of bacterial DNA in the tested samples . Of the 35 PCR-positive samples tested, 33 (94.3%) were positive for Haemophilus, 3 (8.6%) were positive for Streptococcus, and 10 (28.6%) were positive for M . catarrhalis . Ten samples (28.6%) exhibited a mixed infection and were positive for both Haemophilus and M . catarrhalis . Culture was simultaneously performed for all 47 MEE samples . Ten of the 47 MEE samples (21.3%) exhibited bacterial growth . These 10 were PCR positive for bacterial DNA . The remaining 25 PCR-positive samples were negative by culture, thus showing about 53% discordance between PCR results and those of culture . The PCR assay proved to be more sensitive than culture, more rapid, less cumbersome, and more cost-effective than the available PCR-Southern hybridization-based assays.

J Bacteriol, 1998 May, 180(9), 2549 - 55
Regulation of a new cell wall hydrolase gene, cwlF, which affects cell separation in Bacillus subtilis; Ishikawa S et al.; Bacillus subtilis produces a 35-kDa cell wall hydrolase, CwlF, during vegetative growth . The CwlF protein was extracted from B . subtilis cwlB sigD mutant cells and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . N-terminal amino acid sequencing revealed that its sequence is completely identical to that of the internal region of the papQ gene product . Disruption of the papQ gene in the B . subtilis chromosome led to the complete loss of CwlF, indicating that papQ is identical to cwlF . CwlF exhibits high sequence similarity to the p60 proteins of Listeria species, NlpC proteins of Escherichia coli and Haemophilus influenzae, and Enp2 protein of Bacillus sphaericus . The beta-galactosidase activity of the cwlF-lacZ transcriptional fusion and Northern blot analysis of the cwlF gene indicated that the gene is expressed as a monocistronic operon during the exponential growth phase, and primer extension analysis suggested that the cwlF gene is transcribed mainly by EsigmaA RNA polymerase and weakly by EsigmaH RNA polymerase . While the cells of the cwlF-deficient mutant were about twice as long as those of the wild-type strain, the cwlF sigD double mutant cells exhibited extraordinary microfiber formation, in contrast to the filamentation of the sigD mutant . The CwlF production was not affected by the pleiotropic mutations flaD1 and degU32(Hy), which endow cells with the ability of extensive filamentation.

Infect Immun, 1998 May, 66(5), 2272 - 8
Potential of a novel protein, OMP26, from nontypeable Haemophilus influenzae to enhance pulmonary clearance in a rat model; Kyd JM et al.; A major outer membrane protein band of approximately 25 to 27 kDa is commonly observed in strains of Haemophilus influenzae . This study has investigated the potential of a 26-kDa protein (OMP26) from nontypeable H . influenzae (NTHI) as a vaccine candidate . OMP26 was used to immunize rats via intestinal Peyer's patches, followed by an intratracheal boost . Immunization was found to significantly enhance bacterial clearance following pulmonary challenge with both the homologous NTHI strain and a different NTHI strain . Significant levels of anti-OMP26 were found in the serum and bronchoalveolar lavage from immunized rats, and isotypes of immunoglobulin G (IgG) were also measured in serum . Analysis of IgG isotypes present in serum following OMP26-immunization suggest that predominantly a T-helper 1-type response was induced . The OMP26 protein was amino-terminally sequenced and found to have no homology with the P5 of H . influenzae type b P5 or the fimbrin protein of NTHI, both can migrate upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis at similar molecular masses but OMP26 has 100% homology with a segment of the H . influenzae Rd genome . The results of this study suggest that OMP26 may be a suitable vaccine candidate against NTHI infection and warrants continued investigation and characterization.

Infect Immun, 1998 May, 66(5), 2093 - 8
Reduced response to multiple vaccines sharing common protein epitopes that are administered simultaneously to infants; Dagan R et al.; The plethora of newly discovered vaccines implies that, in the future, many vaccines will have to be administered simultaneously to infants . We examined the potential interference with the immune response of several coadministered vaccines containing the same protein component, namely, tetanus toxoid (TT) . Infants simultaneously receiving a tetravalent pneumococcal vaccine conjugated to TT (PncT) and a diphtheria-tetanus-pertussis-poliovirus-Haemophilus influenzae type b-tetanus conjugate vaccine showed significantly lower anti-H . influenzae type b polysaccharide (polyribosylribitol phosphate {PRP}) antibody concentrations than those receiving either a tetravalent pneumococcal vaccine conjugated to diphtheria toxoid or placebo . A dose range study showed that anti-PRP antibody concentrations were inversely related to the TT content of the PncT vaccines administered in infancy . Postimmunization antitetanus antibody concentrations were also affected adversely as the TT content of the coadministered vaccines was increased . This phenomenon, which we believe derives from interference by a common protein carrier, should be taken into account when the introduction of an immunization program including multiple conjugate vaccines is considered.

Infect Immun, 1998 May, 66(5), 1973 - 80
Nasopharyngeal colonization with nontypeable Haemophilus influenzae in chinchillas; Yang YP et al.; Colonization of the nasopharynx by a middle ear pathogen is the first step in the development of otitis media in humans . The establishment of an animal model of nasopharyngeal colonization would therefore be of great utility in assessing the potential protective ability of candidate vaccine antigens (especially adhesins) against otitis media . A chinchilla nasopharyngeal colonization model for nontypeable Haemophilus influenzae (NTHI) was developed with antibiotic-resistant strains . This model does not require coinfection with a virus . There was no significant difference in the efficiency of NTHI colonization between adult (1- to 2-year-old) and young (2- to 3-month-old) animals . However, the incidence of middle ear infection following nasopharyngeal colonization was significantly higher in young animals (83 to 89%) than in adult chinchillas (10 to 30%) . Chinchillas that had recovered either from a previous middle ear infection caused by NTHI or from an infection by intranasal inoculation with NTHI were completely protected against nasopharyngeal colonization with a homologous strain and were found to be the best positive controls in protection studies . Systemic immunization of chinchillas with inactivated whole-cell preparations significantly protected animals not only against homologous NTHI colonization but also partially against heterologous NTHI infection . In all protected animals, significant serum anti-P6 and anti-HMW antibody responses were observed . The outer membrane P6 and high-molecular-weight (HMW) proteins appear to be promising candidate vaccine antigens to prevent nasopharyngeal colonization and middle ear infection caused by NTHI.

Infect Immun, 1998 May, 66(5), 1891 - 7
Synthesis and characterization of lipooligosaccharide-based conjugates as vaccine candidates for Moraxella (Branhamella) catarrhalis; Gu XX et al.; Moraxella (Branhamella) catarrhalis is an important cause of otitis media and sinusitis in children and of lower respiratory tract infections in adults . Lipooligosaccharide (LOS) is a major surface antigen of the bacterium and elicits bactericidal antibodies . Treatment of the LOS from strain ATCC 25238 with anhydrous hydrazine reduced its toxicity 20,000-fold, as assayed in the Limulus amebocyte lysate (LAL) test . The detoxified LOS (dLOS) was coupled to tetanus toxoid (TT) or high-molecular-weight proteins (HMP) from nontypeable Haemophilus influenzae through a linker of adipic acid dihydrazide to form dLOS-TT or dLOS-HMP . The molar ratios of dLOS to TT and HMP conjugates were 19:1 and 31:1, respectively . The antigenicity of the two conjugates was similar to that of the LOS, as determined by double immunodiffusion . Subcutaneous or intramuscular injection of both conjugates elicited a 50- to 100-fold rise in the geometric mean of immunoglobulin G (IgG) to the homologous LOS in mice after three injections and a 350- to 700-fold rise of anti-LOS IgG in rabbits after two injections . The immunogenicity of the conjugate was enhanced by formulation with monophosphoryl lipid A plus trehalose dimycolate . In rabbits, conjugate-induced antisera had complement-mediated bactericidal activity against the homologous strain and heterologous strains of M . catarrhalis . These results indicate that a detoxified LOS-protein conjugate is a candidate for immunization against M . catarrhalis diseases.

Proc Natl Acad Sci U S A, 1998 Mar 31, 95(7), 3720 - 5
Strand compositional asymmetry in bacterial and large viral genomes; Mrazek J et al.; Several bacterial genomes exhibit preference for G over C on the DNA leading strand extending from the origin of replication to the ter-region in the genomes of Escherichia coli, Mycoplasma genitalium, Bacillus subtilis, and marginally in Haemophilus influenzae, Mycoplasma pneumoniae, and Helicobacter pylori . Strand compositional asymmetry is not observed in the cyanobacterium Synechocystis sp . genome nor in the archaeal genomes of Methanococcus jannaschii, Methanobacterium thermoautotrophicum, and Archaeoglobus fulgidus . A strong strand compositional asymmetry is observed in beta-type but not alpha- or gamma-type human herpesviruses featuring G > C downstream of oriL and C > G upstream of oriL . Dinucleotide relative abundances (i.e., dinucleotide representations normalized by the component nucleotide frequencies) are consonant with respect to the leading and lagging strands . Strand compositional asymmetry may reflect on differences in replication synthesis of the leading versus lagging strand, on differences between template and coding strand associated with transcription-coupled repair mechanisms, on differences in gene density between the two strands, on differences in residue and codon biases in relation to gene function, expression level, or operon organization, or on differences in single or context-dependent base mutational rates . The absence of strand asymmetry in the archaeal genomes may reflect the presence of multiple origins of replication.

Scand J Infect Dis, 1997, 29(6), 559 - 63
Antibiotic resistance in Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes in respiratory tract infections in outpatients; Henning C et al.; Sensitivity patterns of Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes were studied prospectively in an outpatient population seeking medical advice for respiratory tract infections (RTI) in the Southern parts of Stockholm . In total, 3,214 nasopharyngeal and 1,907 throat swabs were cultured during January-February 1996 . 32% of the patients had received antibiotics during the previous year . Reduced penicillin sensitivity in S . pneumoniae was rare (1.3%) and only seen in patients treated with antibiotics during the previous 4 months . Beta-lactamase production in H . influenzae was found in 13.4% of patients who had been treated with antibiotics during the last 4 months and in 7.9% of the others . No resistance (< 1%) to erythromycin was seen in S . pyogenes . In this population-based surveillance, the levels of resistance in common respiratory tract pathogens were thus low and correlated to previous antibiotic treatment . Strict indications for antibiotic treatment in uncomplicated RTI are advocated to maintain a low resistance rate . Penicillin is still the drug of choice in patients without frequent recurrences of RTI in a setting similar to the one studied.

Scand J Infect Dis, 1997, 29(6), 555 - 8
Carriage of multiresistant Streptococcus pneumoniae among children attending day-care centres in the Stockholm area; Christenson B et al.; To determine the prevalence of the asymptomatic carriage of drug-resistant Streptococcus pneumoniae (DRSP) by children attending day-care centres in the Stockholm area, nasopharyngeal swabs were cultured from 1129 children and 308 day-care personnel in 36 day-care centres during a 3-week period, from March to April 1995 . Approximately 36% of the children were asymptomatic carriers of S . pneumoniae sensitive to penicillin and other antibiotics . The highest prevalence of nasopharyngeal carriage was found in the 2-year-old group (50%), whereas among the 4-year-old children colonization was observed in 42%, and among the 7-year-old children 21% were asymptomatic carriers of penicillin-sensitive S . pneumoniae . In 2 day-care centres, 4 and 5 children, respectively, were found to have DRSP strains in the nasopharynx . The same serotype of DRSP strain was found among the children attending the same day-care centre . During the same period, none of the staff were found to harbour DRSP in the nasopharynx, but 3% were asymptomatic carriers of penicillin-sensitive S . pneumoniae . The patterns of nasopharyngeal colonization by Haemophilus influenzae, Moraxella catarrhalis and Group A streptococci were also studied in 635 children during the same period . 42% of the nasal cultures yielded Moraxella, 32% H . influenzae and 2% Streptococcus pyogenes.

J Accid Emerg Med, 1998 Mar, 15(2), 72 - 6
The early management of meningococcal disease; Hodgetts TJ et al.; Meningococcal disease is a fulminant infection with an overall mortality of 8% . Mortality is significantly increased with meningococcal septicaemia, particularly when there has been a delay in the diagnosis . The trend from 1985 to 1995 has been an increase in incidence of this disease, and the relative importance of meningococcal disease has also increased following a fall in the incidence of invasive Haemophilus influenzae disease with childhood immunisation . The management of such cases can be complex and time critical . Patients with meningococcal septicaemia often require aggressive resuscitation, including airway support, intravenous colloid, and parenteral antibiotics; hypoglycaemia is also commonly seen, and inotropes may be needed to support the circulation . We examine the treatment strategies in the early management of meningococcal disease and provide an algorithm for use by ambulance personnel, general practitioners, accident and emergency clinicians, and paediatricians . The objective of this algorithm is to ensure that an optimally resuscitated patient is delivered to the definitive care facility.

Vaccine, 1998 Apr, 16(6), 637 - 42
Effect of combination with an acellular pertussis, diphtheria, tetanus vaccine on antibody response to Hib vaccine (PRP-T); Bell F et al.; Acellular pertussis vaccines provide protection against whooping cough with few adverse effects . Their introduction to routine immunisation programmes would be facilitated by their incorporation with other routinely administered vaccines . 262 infants were immunised with an acellular pertussis vaccine containing pertussis toxin and filamentous haemagglutinin, combined with diphtheria and tetanus toxoids . This vaccine was mixed with Haemophilus influenzae type b tetanus toxoid vaccine (PRP-T) so that infants received a single injection at age 2, 3 and 4 months . One month after the third dose the geometric mean titre of Hib IgG antibody was 0.48 microgram ml-1 . Eighty-two percent of infants achieved a titre of 0.15 microgram ml-1, with only 27% achieving 1.0 microgram ml-1 . This combination vaccine induced low Hib antibody responses when compared to other studies in which PRP-T was mixed with acellular or whole-cell pertussis vaccines . The combined vaccine did, however, appear to prime a subset of 35 infants for response to a fourth dose of PRP-T at 13 months of age, with a rise in GMT from 0.21 microgram ml-1 to 36.6 micrograms ml-1 . These data have important implications for the introduction of combination acellular pertussis vaccines.

Vaccine, 1998 Apr, 16(6), 576 - 85
Safety and immunogenicity of a combined five-component pertussis-diphtheria-tetanus-inactivated poliomyelitis-Haemophilus B conjugate vaccine administered to infants at two, four and six months of age; Mills E et al.; Safety, immunogenicity and lot consistency of five-component pertussis combination vaccine (CPDT-IPV//PRP-T) in infants were compared to that of whole cell pertussis combination vaccine (DPT-IPV//PRP-T), as were separate and combined injections of CPDT-IPV and PRP-T . No significant differences in adverse event rates were observed between lots of CPDT-IPV//PRP-T or between separate or combined injections of CPDT-IPV and PRP-T . Minor differences in antibody responses were observed between lots of component pertussis vaccine . Higher concentrations of diphtheria and tetanus antitoxins were induced by separate than by combined injection of CPDT-IPV and PRP-T, but no other differences in immunogenicity were observed . Adverse reactions were more than twice as frequent after whole cell than after component pertussis vaccines . Antibody responses to pertussis toxoid, filamentous hemagglutin and pertactin were significantly greater after component vaccines, while the response to type 3 poliovirus was higher after whole cell vaccine . No significant differences were observed for other vaccine components . CPDT-IPV//PRP-T was safe and immunogenic in infants . Antibody results were similar to those observed in a Swedish field trial that demonstrated CPDT to be 85% effective in preventing clinical pertussis.

J Paediatr Child Health, 1998 Feb, 34(1), 95 - 6
Meningitis due to Haemophilus influenzae type f; Pincus DR et al.; OBJECTIVE: To describe a case of Haemophilus influenzae type f (Hif) meningitis occurring in the H . influenzae type b (Hib) vaccine era . RESULTS: Successful treatment of a case of Hif meningitis in a previously vaccinated 3-year-old girl is described . The outcome was complicated by deafness . No underlying immunosuppression was demonstrated . CONCLUSIONS: Despite the great success of Hib vaccines in reducing invasive disease due to H . influenzae, cases of H . influenzae meningitis continue to occur, caused by less common encapsulated serotypes . Whether there will be an increase in the number of these cases in the vaccine era is unknown and infection due to non-b serotypes requires close monitoring.

Int J Circumpolar Health, 1998 Jan, 57(1), 32 - 9
Nasopharyngeal bacteria found on blood agar plates from healthy children in Greenland; Homoe P et al.; We have systematically studied the aerobic nasopharyngeal bacteria isolated from swabs by unselective subculturing on 5% horse blood agar and chocolate agar in 70 healthy children aged 0-1, 3-5 and 8 years in Nuuk and Sisimiut, Greenland . The purpose was to provide a basis for a better understanding of the infectious pathology and blind antibiotic treatment against potential pathogens thereby improving standard antimicrobial treatment of upper respiratory tract infections (URTI) and otitis media (OM) among children in Greenland . The study serves also as a baseline for future microbiological and immunological research projects . The children were clinically examined for any infectious diseases and a medical history was obtained which allowed for selection of children without a history of severe clinical infection . Nasopharyngeal swabs obtained via the oral route were instantly spread on 5% blood agar and chocolate agar culture plates and incubated aerobically . Subsequently, potentially pathogenic as well as non-pathogenic bacteria were identified by conventional methods . Healthy children in Greenland carry grossly the same aerobic bacterial flora as children in other parts of the world but potentially pathogenic bacteria were found in very high frequency (94%) . Staphylococcus aureus, Streptococcus pneumoniae and Moraxella catarrhalis were found in higher frequencies in the youngest children . Haemophilus influenzae non-b was found in high frequencies in all age groups (67-76%) . H . influenzae type b was carried by 11.4% . Group A streptococci were found more frequently in older children and in children from Sisimiut . Of M . catarrhalis strains 88% produced beta-lactamase . Neisseria meningitidis, Mycoplasma pneumoniae and chlamydiae were not detected at all . The high carrier frequency of potentially pathogenic bacteria in healthy children in Greenland may be related to the high frequency of URTI's and episodes of OM among children in Greenland.

Clin Otolaryngol, 1998 Feb, 23(1), 63 - 6
Bacteraemia during tonsillectomy: a study of the factors involved and clinical implications; Soldado L et al.; Post-tonsillectomy bacteremia is a well-recognized aetiological factor in streptococcal endocarditis, and prophylactic penicillin has been recommended to reduce its incidence in susceptible patients undergoing tonsillectomy . Recent studies have shown a change in the microflora and an increase in the number of penicillin-resistant organisms in the tonsils of patients undergoing tonsillectomy . The aim of this study was to assess the incidence of post-tonsillectomy bacteraemia, to identify the micro-organisms associated with it and to review the suitability of penicillin in prophylactic regimens . The relationship between positive blood cultures and several clinical parameters such as fever, vomiting, pharyngeal discomfort, or dysphagia was also analysed . Of the 102 patients included in the study, 41 (40.1%) had positive post-tonsillectomy blood cultures . Haemophilus influenzae were isolated from 23 (56%) of the positive cultures and Streptococcus viridans in 15 (36.5%) . Twenty-five per cent of H . influenzae produced beta-lactamase and only 30% of streptococci of the viridans group were penicillin-sensitive . Positivity of the blood cultures was not related to fever, discomfort, surgical technique, type of tonsil, or any of the parameters studied . Bacteraemia seems to be related to traction of the tonsil before dissection rather than direct spread of bacteria into the opened vessels . Using a beta-lactamase stable antibiotic instead of penicillin for prophylaxis would be more appropriate.

Int J Tuberc Lung Dis, 1998 Jan, 2(1), 2 - 4
Acute respiratory infections: the forgotten pandemic . Communiqué from the International Conference on Acute Respiratory Infections, held in Canberra, Australia, 7-10 July 1997; IgA1 protease production by bacteria colonizing the upper respiratory tract; Universitatsklinik fur Kinder und Jugendliche, Erlangen, GermanyThirty-eight clinical isolates of Haemophilus influenzae and ten clinical isolates of Streptococcus pneumoniae were examined for IgA1 protease production . A suspension of surface material of each individual strain was incubated with human secretory IgA; IgA1 cleavage products were detected by SDS-PAGE and immunoblotting . The high incidence of IgA1 protease-positive strains (68.4% of the examined H . influenzae and 100% of the examined S . pneumoniae strains) confirms that IgA1 protease activity is a frequent characteristic of these two species . Yet the presence of this enzyme is, if at all, only a minor decisive factor for the induction of symptomatic infections of the upper respiratory tract by IgA1 protease-positive bacteria.

Minerva Med, 1998 Jan-Feb, 89(1-2), 15 - 22
{Correlation between pulmonary pharmacokinetics and pharmacodynamics support the hypothesis of the usefulness of ceftazidime at a single 1g daily dose in the treatment of bacterial exacerbation of chronic obstructive bronchopneumonia with moderate functional damage}; Cazzola M et al.; INTRODUCTION AND BACKGROUND: Experimental studies have shown that cephalosporins have an antibacterial effect in vivo even when their levels are above MIC for only 40-50% of dosing intervals, whereas maximum killing is obtained when concentrations are above MIC for 60-70% of the time . Since most patients treated with antibiotics have neutrophils and other natural defence mechanisms, it is likely that a bacteriostatic effect should be sufficient to induce an effective therapeutic response . METHODS: Given that in the potential sites of lung infection ceftazidime reaches significantly higher levels than the MIC of the most commonplace respiratory pathogens, even 8-12 hours after the administration of 1 g i.m., the authors evaluated the efficacy of treatment of renewed acute episodes of COPD using this antibiotic at a dose of 1 g once a day . In order to do this, 20 outpatients were enrolled in the study, half of whom presented moderate bronchial obstruction (FEV1 = 50-70% of theoretical) whereas the remainder presented marked bronchial obstruction (FEV1 = < 50% of theoretical) . RESULTS: The 10 patients with moderate obstruction at the time of enrollment, who presented Haemophilus influenzae, Streptococcus pneumoniae or Moraxella catarrhalis as causal agents in the sputum (Escherichia coli was only isolated in one patient), showed a marked improvement following treatment with 1 g ceftazidime one a day . A real or presumed eradication of the causal microorganism was observed in all subjects . Treatment with ceftazidime at the dose of 1 g/die once a day was much less effective in patients with marked bronchial obstruction . Treatment was successful in 7 out of 10 subjects, but 2 of them relapsed within 2 weeks . In this second group, Pseudomonas aeruginosa was found in the sputum of 3 patients; one of the patients showed a persistence of the bacterium after ceftazidime treatment, and another presented reinfection 12 days after the end of treatment . The two patients in whom Staphylococcus aureus was isolated did not benefit from ceftazidime treatment at this dosage . One subject who initially presented Streptococcus pneumoniae in his sputum and was then thought to have recovered, underwent a new acute episode caused by Moraxella catarrhalis 2 weeks after the suspension of ceftazidime treatment . CONCLUSIONS: The therapeutic responses observed during this study suggest the possibility of using ceftazidime in a single daily dose of 1 g i.m . to treat those patients with exacerbations of COPD who only present moderately impaired functional symptoms . On the contrary, this type of therapeutic approach must be used with extreme caution in subjects with marked functional damage, although a satisfactory clinical response may be obtained in some cases . However, the small number of patients included in this study does not allow firm conclusions to be drawn . Only a study involving a larger group of patients could provide the necessary information to confirm the hypothesis for treatment put forward by the authors.

Biochem J, 1998 May 1, 331 ( Pt 3), 897 - 904
Periplasmic nitrate-reducing system of the phototrophic bacterium Rhodobacter sphaeroides DSM 158: transcriptional and mutational analysis of the napKEFDABC gene cluster; Reyes F et al.; The phototrophic bacterium Rhodobacter sphaeroides DSM 158 is able to reduce nitrate to nitrite by means of a periplasmic nitrate reductase which is induced by nitrate and is not repressed by ammonium or oxygen . Recently, a 6.8 kb PstI DNA fragment carrying the napABC genes coding for this periplasmic nitrate-reducing system was cloned {Reyes, Roldan, Klipp, Castillo and Moreno-Vivian (1996) Mol . Microbiol . 19, 1307-1318} . Further sequence and genetic analyses of the DNA region upstream from the napABC genes reveal the presence of four additional nap genes . All these R . sphaeroides genes seem to be organized into a napKEFDABC transcriptional unit . In addition, a partial open reading frame similar to the Azorhizobium caulinodans yntC gene and the Escherichia coli yjcC and yhjK genes is present upstream from this nap gene cluster . The R . sphaeroides napK gene codes for a putative 6.3 kDa transmembrane protein which is not similar to known proteins and the napE gene codes for a 6.7 kDa transmembrane protein similar to the Thiosphaera pantotropha NapE . The R . sphaeroides napF gene product is a 16.4 kDa protein with four cysteine clusters that probably bind four {4Fe-4S} centres . This iron-sulphur protein shows similarity to the NapF and NapG proteins of E . coli and Haemophilus influenzae . Finally, the napD gene product is a 9.4 kDa soluble protein which is also found in E . coli and T . pantotropha . The 5' end of the nap transcript has been determined by primer extension, and a sigma70-like promoter has been identified upstream from the napK gene . The same transcriptional start site is found for cells growing aerobically or anaerobically with nitrate . Different mutant strains carrying defined polar and non-polar insertions in each nap gene were constructed . Characterization of these mutant strains demonstrates the participation of the nap gene products in the periplasmic nitrate reduction in R . sphaeroides.

Antimicrob Agents Chemother, 1998 Apr, 42(4), 785 - 8
In vivo activity of HSR-903, a new fluoroquinolone, against respiratory pathogens; Yoshizumi S et al.; The in vivo activity of HSR-903, a new fluoroquinolone, against major bacteria which cause respiratory tract infections was evaluated . HSR-903 was active against experimental respiratory tract infections in mice challenged with penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae and Haemophilus influenzae strains . Treatment with HSR-903 reduced the bacterial numbers in infected murine lungs . In accord with the pulmonary clearance results, the rates of survival for mice treated with HSR-903, sparfloxacin, levofloxacin, ciprofloxacin, and benzylpenicillin were 50, 30, 10, 0, and 0%, respectively, 14 days after being infected with penicillin-resistant S . pneumoniae . A pharmacokinetic study with pneumonic mice showed that the levels of HSR-903 in the lungs were seven to eight times higher than those in the plasma . These results indicate that clinical studies of HSR-903 against respiratory tract infections may be warranted.

Antimicrob Agents Chemother, 1998 Apr, 42(4), 772 - 8
Effect of dirithromycin on Haemophilus influenzae infection of the respiratory mucosa; Rutman A et al.; Macrolides have properties other than their antibiotic action which may benefit patients with airway infections . We have investigated the effect of dirithromycin (0.125 to 8.0 microg/ml) on the interaction of Haemophilus influenzae with respiratory mucosa in vitro using human nasal epithelium, adenoid tissue, and bovine trachea . Dirithromycin did not affect the ciliary beat frequency of the nasal epithelium or the transport of mucus on bovine trachea, but dirithromycin (1 microg/ml) did reduce the slowing of the ciliary beat frequency and the damage to the nasal epithelium caused by H . influenzae broth culture filtrate . Amoxicillin (2 microg/ml) did not reduce the effects of the H . influenzae broth culture filtrate . H . influenzae infection of the organ cultures for 24 h caused mucosal damage and the loss of ciliated cells . Bacteria adhered to damaged epithelium and to a lesser extent to mucus and unciliated cells . Incubation of H . influenzae with dirithromycin at sub-MICs (0.125 and 0.5 microg/ml) prior to infection of the organ cultures did not reduce the mucosal damage caused by bacterial infection . By contrast, incubation of adenoid tissue with dirithromycin (0.125 to 1.0 microg/ml) for 4 h prior to assembling the organ culture reduced the mucosal damage caused by subsequent H . influenzae infection by as much as 50% . The number of bacteria adherent to the mucosa was reduced, although the tissue that had been incubated with dirithromycin (0.125 and 0.5 microg/ml) did not inhibit bacterial growth . This was achieved by a reduction in the amount of damaged epithelium to which H . influenzae adhered and a reduction in the density of bacteria adhering to mucus . We conclude that dirithromycin at concentrations achievable in vivo markedly reduces the mucosal damage caused by H . influenzae infection due to a cytoprotective effect.

Antimicrob Agents Chemother, 1998 Apr, 42(4), 729 - 33
Cefepime versus ceftriaxone for empiric treatment of hospitalized patients with community-acquired pneumonia . The Cefepime Study Group; Zervos M et al.; Effective empiric treatment of pneumonia requires antibiotic coverage against gram-negative and gram-positive pathogens, including drug-resistant isolates . We compared the safety and efficacy of intravenous (i.v.) cefepime (2 g administered every 12 h) to those of i.v . ceftriaxone (1 g administered every 12 h) for the empiric treatment of hospitalized patients with community-acquired pneumonia . Of the 115 patients randomized to the study, 86 (cefepime recipients, n = 40; ceftriaxone recipients, n = 46) were evaluated for clinical efficacy (clinically evaluated patients) . Favorable clinical outcomes (cure or improvement) were comparable among clinically evaluated patients in the cefepime and ceftriaxone treatment arms (95.0 versus 97.8%, respectively; 95% confidence interval for treatment difference {data for ceftriaxone group minus data for cefepime group}, -5.1 to +10.8%) . The most common bacteria isolated from patients in both treatment groups were Streptococcus pneumoniae, Haemophilus influenzae, and Staphylococcus aureus . In clinically evaluated patients with a microbiologic response, all (100%) of the 32 pathogens from cefepime-treated patients and 97.4% (38 of 39) of the pathogens from ceftriaxone-treated patients were eradicated (documented or presumed eradication) . The one persistent infection in the ceftriaxone group was caused by Pseudomonas fluorescens . Both treatments were well tolerated . Our data thus suggest that cefepime and ceftriaxone have comparable safety and efficacy for the treatment of pneumonia in hospitalized patients.

Jpn J Antibiot, 1998 Jan, 51(1), 1 - 10
{Antimicrobial activities of cefcapene against clinical isolates from respiratory tract infections of outpatients}; Ishihara R et al.; In order to evaluate antimicrobial activity of cefcapene (CFPN), minimum inhibitory concentrations (MICs) of CFPN and reference drugs were determined against clinical isolates from respiratory tract infection of out patients that were obtained in our laboratory from January to June of 1997 . The results are summarized as follows; 1 . The MIC90 of CFPN against penicillin (PC)-susceptible Streptococcus pneumoniae (PSSP) was equal to those of benzylpenicillin (PCG), ampicillin (ABPC) and cefditoren (CDTR), and was lower than those of cefaclor (CCL), cefdinir (CFDN) and erythromycin (EM) . 2 . The MIC90 of CFPN against PC-intermediate S . pneumoniae (PISP)/PC-resistant S . pneumoniae (PRSP) was equal to that of CDTR, and was lower than those of PCG, ABPC, CCL, CFDN and EM . CFPN showing strong antimicrobial activities against PISP . 3 . CFPN showed strong antimicrobial activities against beta-lactamase producing and non-producing Haemophilus influenzae . The MIC90 of CFPN was stronger than those of ABPC, CCL, CFDN and EM, and was approximately equal to that of CDTR . CFPN also showed strong antimicrobial activities against strains which did not produce any beta-lactamase and were resistant to CCL with MIC of > or = 25 micrograms/ml . 4 . Antimicrobial activities of CFPN against Moraxella subgenus Branhamella catarrhalis was stronger than that of ABPC and CCL, though the MIC90 of CFPN was rather high, 3.13 micrograms/ml . 5 . CFPN showed strong antimicrobial activities against PISP and beta-lactamase producing H . influenzae, and also against the CCL-resistant H . influenzae indicative mutations of penicillin-binding proteins (PBPs) . From those results, cefcapen-pivoxil was found to be clinically effective against community acquired respiratory tract infection.

IEEE Trans Biomed Eng, 1998 Apr, 45(4), 429 - 39
Systematic method for determining intravenous drug treatment strategies aiding the humoral immune response; Rundell AE et al.; This paper delineates a systematic method for determining "optimal" intravenous drug delivery strategies for patients having illnesses that primarily evoke a humoral immune response and are treatable by antibiotics . The method derives from a nonlinear, distributed predator-prey model that captures the dominant antigen and antibody interaction . This model is developed from relevant physiology, past predator-prey-type modeling work, available data, and pertinent parameter identification . Embedding this predator-prey model into a larger class of uncertain systems, by a finite dimensional approximation and a transformation to a linear fractional representation, enables the application of robust control based on linear matrix inequality optimization techniques . The optimization problem is solved by minimizing an upper bound on a measure of the total drug delivered subject to patient recovery (stability to healthy equilibrium state) . Specifically, the paper addresses the treatment of Haemophilus influenzae through modeling, controller development, and simulations of infected adult patients subjected to typical and proposed intravenous antibiotic treatments . Through simulations the proposed intravenous drug strategy shortens patient recovery time, lowers peak drug concentrations and decreases the total drug administered when compared to standard antibiotic strategies.

J Bacteriol, 1998 Apr, 180(8), 2087 - 92
The periplasmic, group III catalase of Vibrio fischeri is required for normal symbiotic competence and is induced both by oxidative stress and by approach to stationary phase; Visick KL et al.; The catalase gene, katA, of the sepiolid squid symbiont Vibrio fischeri has been cloned and sequenced . The predicted amino acid sequence of KatA has a high degree of similarity to the recently defined group III catalases, including those found in Haemophilus influenzae, Bacteroides fragilis, and Proteus mirabilis . Upstream of the predicted start codon of katA is a sequence that closely matches the consensus sequence for promoters regulated in Escherichia coli by the alternative sigma factor encoded by rpoS . Further, the level of expression of the cloned katA gene in an E . coli rpoS mutant is much lower than in wild-type E . coli . Catalase activity is induced three- to fourfold both as growing V . fischeri cells approach stationary phase and upon the addition of a small amount of hydrogen peroxide during logarithmic growth . The catalase activity was localized in the periplasm of wild-type V . fischeri cells, where its role could be to detoxify hydrogen peroxide coming from the external environment . No significant catalase activity could be detected in a katA null mutant strain, demonstrating that KatA is the predominately expressed catalase in V . fischeri and indicating that V . fischeri carries only a single catalase gene . The catalase mutant was defective in its ability to competitively colonize the light organs of juvenile squids in coinoculation experiments with the parent strain, suggesting that the catalase enzyme plays an important role in the symbiosis between V . fischeri and its squid host.

Dev Biol Stand, 1998, 92, 79 - 87
PCPP as a parenteral adjuvant for diverse antigens; Payne LG et al.; The adjuvanticity of the phosphazene polymer, poly{di(carboxylatophenoxy) phosphazene} (PCPP) was examined with a diverse collection of immunogens . PCPP proved to be a potent adjuvant for trivalent influenza virus vaccine, tetanus toxoid, hepatitis B surface antigen, herpes simplex virus glycoprotein gD2 and the capsular polysaccharide, polyribosylribitolphosphate, from Haemophilus influenzae type b . Taken together these results clearly demonstrate the general utility of PCPP as an adjuvant . Furthermore, PCPP was a superior adjuvant at least with TT compared to similar negatively charged polyanions, polymethylacrylic acid and polyacrylic acid.

Dev Biol Stand, 1998, 92, 63 - 78
Biodegradable polymer microspheres as vaccine adjuvants and delivery systems; Gupta RK et al.; Though vaccination has been the most cost-effective way of controlling infectious diseases, the logistics of delivering at least two to three doses of conventional vaccines for primary immunization to achieve protection are difficult and compliance is frequently inadequate, particularly in developing countries . In recent years biodegradable polymer microspheres have received much attention for the purposes of controlled release of antigens, (i) to reduce the number of doses needed for primary immunization to as few as a single dose and (ii) to target an antigen to microfold cells on mucosal surfaces after oral administration or to antigen-presenting cells after parenteral inoculations . A variety of vaccine antigens have been encapsulated in microspheres usually composed of poly (lactic/glycolic) acid (PLGA) . Based on the size of the microspheres, molecular weight of polymer and ratio of lactic to glycolic acid in the polymer, the antigen may be targeted to various cells of the immune system or it may form a depot at the site of injection, allowing the slow release of the antigen for extended periods . Additionally, another adjuvant may be incorporated inside microspheres together with the antigen, further enhancing or modulating the immune response to the desired type . The major problems in developing controlled-release vaccines include instability of vaccine antigens during micro-encapsulation, storage and subsequent hydration . We encapsulated tetanus toxoid (TT) and Haemophilus influenzae type b capsular polysaccharide conjugated to TT (Hib-T) inside PLGA microspheres and evaluated the antibody levels in mice . A single injection of these micro-encapsulated vaccines elicited high antibody levels which persisted for several months . The antibody levels were similar or superior to those elicited by conventional formulations of AIPO4-adsorbed TT or soluble Hib-T conjugate vaccine.

Acta Microbiol Immunol Hung, 1997, 44(4), 355 - 9
A new selective method for isolation of Haemophilus species; Csukas Z et al.; Chocolate agar with teicoplanin disk (30 micrograms) was used for the isolation of Haemophilus strains . Fifty strains of 3 Haemophilus species grew as well in the inhibition zones of teicoplanin disks as on teicoplanin containing selective plates, whereas Gram-positive bacteria failed to form colonies . This selective method proved especially advantageous when Haemophilus strains were isolated from mixed bacterial cultures of 665 specimens.

Int J Antimicrob Agents, 1997 Jan, 9(3), 141 - 6
A short (3-day) course of azithromycin tablets versus a 10-day course of amoxycillin-clavulanic acid (co-amoxiclav) in the treatment of adults with lower respiratory tract infections and effects on long-term outcome; Hoepelman IM et al.; The efficacy and safety of a 3-day regimen of azithromycin prescribed in the new tablet form and of a 10-day regimen of amoxycillin clavulanic acid (co-amoxiclav, Augmentin) were compared in patients with acute lower respiratory tract infections . Of the 144 enrolled patients, 123 had a Type 1 acute exacerbation of chronic bronchitis (AECB), three patients had pneumonia, and 18 had purulent bronchitis . Treatment was successful, defined as cure or major improvement on day 14, in 59/62 (95%) patients in the azithromycin treatment group compared with 54/61 (90%) patients in the co-amoxiclav . At 30 days, the incidence of success was 77% (48/62) in the azithromycin treated group, compared with 66% (40/61) of co-amoxiclav-treated patients . At 60 days, incidences were 66% (41/62) and 59% (36/61), respectively . Several pathogens were isolated: Haemophilus influenzae in 21 patients (minimum inhibitory concentration (MIC) range for azithromycin 0.12-4 mg/l; co-amoxiclav 0.25-4 mg/l); Streptococcus pneumoniae in nine (MIC azithromycin < or = 0.06 > or = 256 mg/l; co-amoxiclav < or = 0.06-1 mg/l); and Moraxella catarrhalis in 11 (MIC azithromycin < or =0.06-2 mg/l; co-amoxiclav < or = 0.06-0.5 mg/l) . Microbiological response rates were comparable . A significant correlation between clinical and microbiological cure was found (p = 0.02, power 0.6) . In 15 (10%) patients, positive serology for viruses or atypical pathogens was found . In the co-amoxiclav-treatment group, 24 patients had mild adverse events (12 diarrhoea), compared with 27 treated with azithromycin (p = 0.47) . It is concluded that a 3-day regimen of azithromycin prescribed as tablets is as clinically and microbiologically effective as a 10-day regimen of co-amoxiclav in the treatment of acute lower respiratory tract infections . Moreover, since the percentage of viral infections was low and a significant correlation between microbiological and clinical cure was found, this study shows that clinical symptoms can be used to establish which patients with AECB (Type 1) should be treated with antimicrobial agents.

Chest, 1998 Apr, 113(4 Suppl), 249S - 255S
The value of antibiotics and the outcomes of antibiotic therapy in exacerbations of COPD; Grossman RF; COPD is the fifth leading cause of death in the United States, and acute respiratory infections account for a significant proportion of all primary care visits . Approximately one half of all exacerbations of COPD can be attributed to bacterial infection, and antibiotic therapy has been demonstrated to improve clinical outcomes and hasten clinical and physiologic recovery . The major pathogen continues to be Haemophilus influenzae, and resistance to beta-lactam antibiotics such as ampicillin can be expected in 20 to 40% of isolated strains . Certain high-risk patients, in whom the cost of clinical treatment failure is high, can be identified by simple clinical criteria . Patients with significant cardiopulmonary comorbidity, frequent purulent exacerbations of COPD, advanced age, generalized debility, malnutrition, chronic corticosteroid administration, long duration of COPD, and severe underlying lung function tend to fail therapy with older drugs, such as ampicillin, and early relapse can be expected . Treatment directed toward resistant pathogens with potent bactericidal drugs may be expected to lead to improved clinical outcomes and overall lower costs, particularly if hospital admissions and respiratory failure can be prevented . Future studies examining the role of antibiotics should enroll these high-risk patients to determine if new therapies have significant clinical, quality-of-life, and economic advantages over older agents.

Chest, 1998 Apr, 113(4 Suppl), 242S - 248S
The role of infection in COPD; Wilson R; Clinical studies of acute exacerbations of COPD are difficult because of the heterogeneous nature of COPD, diffuse symptoms that can vary spontaneously, and difficulties in defining clinical response both in the short and long term . The role of bacterial infection, and thus use of antibiotics, in COPD is controversial . The available evidence shows that bacterial infection has a significant role in acute exacerbations, but its role in disease progression is less certain . Upper respiratory tract commensals, such as nontypable Haemophilus influenzae, cause most bronchial infections by exploiting deficiencies in the host defenses . Some COPD patients are chronically colonized by bacteria between exacerbations, which represents an equilibrium in which the numbers of bacteria are contained by the host defenses but not eliminated . When an exacerbation occurs, this equilibrium is upset and bacterial numbers increase, which incites an inflammatory response . Neutrophil products can further impair the mucosal defenses, favoring the bacteria, but if the infection is overcome, symptoms resolve . However, if the infection persists, chronic inflammation may cause lung damage . About half of exacerbations involve bacterial infection, but these patients are not easy to differentiate from those who are uninfected, which means that antibiotics have to be given more often than is strictly necessary . Further research is needed to characterize those patients in whom bacterial infection has a more important role.

Pediatrics, 1998 Apr, 101(4 Pt 1), 617 - 9
Acute otitis media in children with bronchiolitis; Andrade MA et al.; OBJECTIVE: We investigated the prevalence and the etiology of acute otitis media (AOM) in children with bronchiolitis to determine whether AOM in such children is due entirely or mainly to respiratory syncytial virus (RSV), in which case routine antimicrobial treatment would not be appropriate . METHODS: The study group consisted of children aged 2 to 24 months with bronchiolitis . In patients with AOM at entry, nasal washings for RSV enzyme-linked immunosorbent assay were obtained, and Gram-stained smear, bacterial culture, and reverse transcriptase polymerase chain reaction to detect the presence of RSV were performed on middle-ear aspirates . Patients without AOM were reevaluated at 48 to 72 hours, 8 to 10 days, and 18 to 22 days . RESULTS: Forty-two children with bronchiolitis were enrolled . Sixty-two percent had AOM at entry or developed AOM within 10 days . An additional 24% had or eventually developed otitis media with effusion . Only 14% remained free of both AOM and otitis media with effusion throughout the 3-week observation period . All patients with AOM had 1 or more bacterial pathogens isolated from one or both middle-ear aspirates . Of 33 middle-ear aspirates, Streptococcus pneumoniae was isolated in 15, Haemophilus influenzae in 8, Moraxella catarrhalis in 8, and Staphylococcus aureus in 2 . Two middle-ear aspirates yielded 2 pathogens each; 2 aspirates had no growth . RSV was identified in 17 (71%) of 24 patients with AOM . CONCLUSION: Bacterial AOM is a complication in most children with bronchiolitis . Accordingly, in patients with bronchiolitis and associated AOM, antimicrobial treatment is indicated.

Pediatrics, 1998 Apr, 101(4 Pt 1), 604 - 11
Safety and immunogenicity of heptavalent pneumococcal vaccine conjugated to CRM197 in United States infants; Rennels MB et al.; OBJECTIVE: To determine the safety and immunogenicity of heptavalent pneumococcal saccharide vaccine (serotypes 4, 6B, 9V, 14, 18C, 19F, 23F) individually conjugated to CRM197 (PNCRM7), administered at 2, 4, 6, and 12 to 15 months of age . DESIGN: Two hundred twelve healthy 2-month-old infants were equally randomized to receive four consecutive doses of PNCRM7 or an investigational meningococcal group C conjugate vaccine, which served as a control . Concomitantly administered routine vaccines were oral polio vaccine and combined diphtheria toxoid, tetanus toxoid, and whole cell pertussis vaccine/Haemophilus influenzae type b vaccine consisting of capsular oligosaccharides conjugated to CRM197 (DTP/HbOC) at 2, 4, and 6 months, and either measles-mumps-rubella vaccine or HbOC at 12 to 15 months . Active safety surveillance was conducted for 3 days after each dose . Antibody concentrations to each of the 7 pneumococcal serotypes were measured by enzyme-linked immunosorbent assay prevaccination, after doses two and three, prebooster, and postbooster . RESULTS: Significantly fewer children experienced local reactions at the PNCRM7 injection site than at the DTP/HbOC site . There was no increase in the incidence or severity of local reactions at the PNCRM7 site with increasing doses of vaccine . Mild to moderate postvaccination fever was common in both the PNCRM7 and control vaccine groups, however DTP/HbOC was administered concurrently . All 7 vaccine serotypes were immunogenic . The kinetics of the immune responses were serotype-specific . After three doses of PNCRM7, between 92% to 100% of children had >/=0.15 microg/mL of antibody, and 51% to 90% achieved a level of >/=1 microg/mL against specific serotypes . A booster dose of PNCRM7 resulted in a brisk anamnestic response to all 7 vaccine serotypes, demonstrating effective stimulation of T-cell memory by the primary series of vaccinations . CONCLUSION: Primary immunization followed by a booster dose of PNCRM7 seemed to be acceptably safe and resulted in significant rises in antibody to all 7 serotypes . Implications . Studies to assess vaccine efficacy of PNCRM7 for prevention of systemic disease, nasopharyngeal colonization, and acute otitis media are in progress . If PNCRM7 proves to be protective, there is the potential to prevent up to 85% of invasive pneumococcal disease occurring in US children.

Pediatrics, 1998 Apr, 101(4 Pt 1), 597 - 603
Three-year follow-up of vaccine response in extremely preterm infants; Khalak R et al.; OBJECTIVE: To assess whether the adequate antibody response observed in former extremely premature infants after the primary series of immunizations is sustained after the first booster vaccines . SUBJECTS AND METHODS: Sixteen former extremely premature (<29 weeks, <1000 g at birth) and 17 former full-term (>37 weeks) infants had sera obtained for antibody titer measurement at 3 to 4 years of age . All had received the primary series and first booster vaccines for diphtheria, pertussis, tetanus, polio, and Haemophilus influenzae type b . Twelve preterm and 14 full-term children had completed the hepatitis B vaccine series . RESULTS: At 3 to 4 years of age, former preterm and full-term children had similar geometric mean titer (GMT) values of antibodies to tetanus, diphtheria, and pertussis . Preterm children had a lower GMT value of Haemophilus polyribosylribitol phosphate (PRP) antibody than did full-term children (0.99 vs 3.06 microg/mL) . Fifty percent of preterm and 88% of full-term children had PRP antibody >1.0 microg/mL; 100% of preterm and 94% of full-term children had anti-PRP titers >0.15 microg/mL . GMT values of neutralizing antibodies to polio serotypes 1 and 2 were similar, with 94% to 100% of both groups above protective levels (>/=1:8) . The difference in GMT values of polio serotype 3 approached significance (29 vs 73); fewer preterm children had protective titer values (75% vs 100%) . Among children vaccinated against hepatitis B, 75% of preterm and 71% of full-term children were protected (10 mIU/mL) . CONCLUSIONS: Preterm children immunized at the recommended chronological ages displayed antibody responses similar to those for full-term children for most immunizing antigens . Responses to PRP and polio serotype 3 were less robust than those of full-term children.

Pharmacotherapy, 1998 Mar-Apr, 18(2), 345 - 57
Rifampin, a useful drug for nonmycobacterial infections; Vesely JJ et al.; Rifampin has clinical efficacy against a wide variety of organisms, including Staphylococcus aureus, Legionella pneumophila, group A Streptococcus, Brucella sp, Haemophilus influenzae, and Neisseria meningitidis, as well as in vitro activity against penicillin-resistant Streptococcus pneumoniae, Neisseria gonorrhoeae, Chlamydia trachomatis, Haemophilus ducreyi, and many gram-negative rods . Rifampin is a useful drug for several types of bacterial infections because of its broad spectrum of activity, excellent tissue penetration, and low side effect profile . In combination with other antibiotics, it may be effective when conventional therapies are not.

J Korean Med Sci, 1998 Feb, 13(1), 60 - 4
The causative organisms of bacterial meningitis in Korean children, 1986-1995; Kim KH et al.; Bacterial meningitis remains a serious cause of morbidity and mortality in childhood . Epidemiologic investigations have shown variability in disease risks among different populations and races . In Korea, however, basic epidemiologic information on bacterial meningitis in children is limited . The main purpose of this study was to analyze bacteriologically proven meningitis cases in terms of the relative frequency of causative organisms, mortality rate, and age distribution beyond the neonatal period . Data was obtained from the hospital records who had been diagnosed with bacterial meningitis at 13 general or university hospitals from 1986 through 1995 . The patients had at least one positive CSF culture for bacteria . Of 140 cases of CSF culture-proven bacterial meningitis, 46.4% was < or =1 year, 62.1% was < or =2 years, 81.4% was < or =5 years cumulatively . Streptococcus pneumoniae was the most common bacteria responsible for 48 (35.0%) of all cases regardless of age, followed by Haemophilus influenzae for 48 (34.3%) and Neisseria meningitidis for 8 (6.4%) patients . The case fatality rate was 20.0%, 17.1%, and 16.7% for N . meningitidis, S . pneumoniae, and H . influenzae, respectively . In conclusion, the most common organisms of culture-proven bacterial meningitis in the last 10 years have been S . pneumoniae, H . influenzae, and N . meningitidis in order of frequency . Further study should be extended to nation-wide epidemiologic evaluation to show the incidence of bacterial meningitis caused by these three important organisms.

J Cardiovasc Surg (Torino), 1998 Feb, 39(1), 113 - 6
Empyema in children; Sarihan H et al.; Empyemas develop following bacterial pneumonias, thoracic trauma and surgery which are still among the common diseases, causing illness and death throughout the developing world . With the advent of potent antibiotics the mortality of empyema has been drastically reduced . In this study 52 patients (29 boys and 23 girls) with thoracic empyema were evaluated retrospectively . In this series the causes of empyema were postpneumonic in 50 patients, esophageal anastomotic leak in one patient, and thoracic trauma in one patient . The diagnosis was suspected clinically and by the finding of a pleural effusion on chest roentgenogram . Definitive diagnosis was confirmed by pleural aspiration which pus was obtained . Responsible organisms included; Staphylococcus aureus, Streptococcus pneumonia, Haemophilus influenza, pseudomonas, and Klebsiella . The most common is Staphylococcus aureus . The patients were treated in various ways; 14 patients were treated with antibiotics and thoracentesis, 38 patients were treated with a closed tube thoracostomy . Eight of 38 patients had the chest tube converted to an open empyema tubes for long term management . Fourteen of 38 patients developed abcess formation . Nine of 14 patients were treated with computed tomography guided catheter placement, five patients encountered thoracotomy and decortication . In this article, appropriate treatment and result of long-term follow-up of empyema were evaluated.

Eur J Pediatr, 1998 Mar, 157(3), 208 - 14
Immunogenicity and reactogenicity of a Haemophilus influenzae type b tetanus conjugate vaccine when administered separately or mixed with concomitant diphtheria-tetanus-toxoid and acellular pertussis vaccine for primary and for booster immunizations; Schmitt HJ et al.; With an increasing number of new vaccines available for routine childhood immunization, combination vaccines are needed in order to maintain or achieve a high compliance with recommended immunization programmes . In a prospective, randomized, comparative, multi-centre study, 822 healthy infants were enrolled to receive three doses of either a candidate or a commercially available Haemophilus influenzae type b (Hib) vaccine concomitantly with diphtheria-, tetanus- acellular pertussis (DTaP) vaccine . Study subjects were randomly allocated to one of the following groups: (1) separate, or (2) mixed injection of DTaP and candidate Hib vaccine, or (3) separate injection of DTaP and commercial Hib vaccine . One year later the first 189 study subjects received either separate or mixed injections of the same Hib and DTaP vaccines as booster doses . Evaluation of reactogenicity was based on diary cards completed by parents . Immunogenicity was documented by measuring IgG antibody concentrations in serum samples taken before and 4 weeks after primary and booster vaccination . No serious adverse events occurred and most local and systemic reactions were mild to moderate . Booster doses were more reactogenic than primary doses with all groups . Antibody concentrations against pertussis antigens were similar to those seen with DTaP alone . All but one subject had protective antibody concentrations against diphtheria and tetanus . Primary immune response to the Hib vaccine was significantly lower in the group receiving the mixed Hib-DTaP vaccine, however, > or = 95% of vaccinees had anti-Hib antibody concentrations > or = 0.15 microg/ml and there was a marked booster response (> 100-fold) in all groups . CONCLUSIONS: Mixing DTaP and Hib vaccines for primary immunization caused a decrease in anti-Hib antibody response, although after primary immunization as after booster doses, all subjects showed antibody concentrations considered to be protective for invasive Hib disease . Mixing of the vaccines did not result in increased reactogenicity.

Trop Med Int Health, 1998 Feb, 3(2), 95 - 9
Immunogenicity of Haemophilus influenzae-diphtheria CRM197 protein conjugate vaccine (HbOC) in Libyan infants; Hadida M et al.; The immunogenicity of the HbOC, a Haemophilus influenzae type b conjugate vaccine, was evaluated in a randomized clinical trial of Arab children resident in Tripoli . The HbOC vaccine was given as part of a three-dose series at 2, 3 and 4 months of age together with hepatitis B, OPV and DPT to 90 children . Anti-H . influenzae antibody levels were compared with 81 infants receiving hepatitis B, OPV and DPT but not the HbOC vaccine . The immunogenicity and safety of HbOC was as high as that observed in industrialised countries . There were no major complications, and fever and temporary local discomfort were observed in fewer than approximately 2% of the infants . Infants receiving the HbOC vaccine had an increase in Hib antibodies with only one dose . Geometric mean anti-Hib antibody levels were 0.41, 1.36 and 2.91 mg/ml after one, two and three doses . After two doses, all children had antibody levels above 0.20 mg/ml and the lowest antibody concentration was 0.80 mg/ml . Antibody levels in our children are similar to those observed in Europe and the USA and it is thus likely that HbOC will provide good clinical protection in this population . As most of the children develop antibody titres above or near 1 mg/ml, it is likely that they are protected even with two doses of the vaccine . The anti-Hib antibody levels observed are similar to those in studies from Europe where hepatitis B vaccine is not routinely given.

J Child Neurol, 1998 Mar, 13(3), 113 - 9
Acute-phase neurologic complications of Haemophilus influenzae type b meningitis: association with developmental problems at school age; Taylor HG et al.; The purposes of this study were to describe the incidence of acute-phase neurologic complications in a sample of 126 children with Haemophilus influenzae type b meningitis, and to determine if these complications were associated with higher rates of learning and behavior problems at school age . Risks were assessed by comparing rates of adverse psychoeducational outcomes in the 53 children in the sample with complications to corresponding outcome rates in the 67 children who were free of neurologic complications and who did not have abnormal electroencephalograms (EEGs) or computed tomographic (CT) scans . Comparisons were made by means of logistic regression analysis . Twenty-nine children (23% of the sample) had seizures, 16 (13%) were comatose or obtunded, 15 (12%) had sensorineural hearing loss, 8 (6%) had hemiparesis, and 7 (6%) had cranial nerve deficits other than hearing loss . Relative to children without complications, those with complications had higher rates of grade repetition and substandard performance on neuropsychological and achievement testing . Adverse outcomes, however, consisted primarily of more subtle cognitive and learning problems; only two of the children in the sample obtained prorated IQ scores below 70 . Sequelae were associated with persistent neurologic deficits and bilateral hearing loss, as well as with transient symptoms including seizures, coma, and hemiparesis . While study findings argue against adverse consequences for the vast majority of children treated for this disease, the results clarify learning and behavior outcomes and indicate which children are at greatest risk.

Mol Microbiol, 1998 Mar, 27(5), 1021 - 9
Identification of the Chi site of Haemophilus influenzae as several sequences related to the Escherichia coli Chi site; Sourice S et al.; The Escherichia coli Chi site 5'-GCTGGTGG-3' modulates the activity of the powerful dsDNA exonuclease and helicase RecBCD . Genome sequence analyses revealed that Chi is frequent on the chromosome and oriented with respect to replication on the E . coli genome . Chi is also present much more frequently than predicted statistically for a random 8-mer sequence . Although it is assumed that Chi is ubiquitous, there is virtually no proof that its features are conserved in other microorganisms . We therefore identified and analysed the Chi sequence of an organism for which the full genome sequence was available, Haemophilus influenzae . The biological test we used is based on our finding that rolling circle plasmids provide a specific substrate for RecBCD analogues in different microorganisms . Unexpectedly, several related sequences, corresponding to 5'-GNTGGTGG-3' and 5'-G(G/C)TGGAGG-3', showed Chi activity . As in E . coli, the H . influenzae Chi sites are frequent on the genome, which is in keeping with the need for frequent Chi sites for dsDNA break repair of chromosomal DNA . Although statistically over-represented, this feature is less marked than that of the E . coli Chi site . In contrast to E . coli, the H . influenzae Chi motifs are only slightly oriented with respect to the replication strand . Thus, although Chi appears to have a highly conserved biological role in attenuating exonuclease activity, its sequence characteristics and statistical representation on the genome may differ according to the particular features of the host.

J Infect Dis, 1998 Apr, 177(4), 1112 - 5
Antibody avidity as a surrogate marker of successful priming by Haemophilus influenzae type b conjugate vaccines following infant immunization; Goldblatt D et al.; Evaluation of the new generation of conjugate vaccines is hampered by the absence of reliable surrogate markers of immunologic memory . Memory responses are characterized by rapid production of relatively high-avidity antibody; thus, a solid-phase ELISA was adapted for the measurement of anti-Haemophilus influenzae type b (Hib) IgG avidity . In a cohort of infants vaccinated at 2, 3, and 4 months of age with Hib conjugate vaccines, avidity increased in the period following vaccination, while antibody titer fell . After a booster dose at 1 year of age, both antibody titer and avidity increased . In a cohort with anti-Hib IgG <1.0 microg/mL following primary immunization, antibody avidity after booster was low, indicating an absence of priming . Antibody avidity may help distinguish, in persons with low antibody titers, between those who are primed for memory and those who are not.

Microb Pathog, 1998 Feb, 24(2), 75 - 87
Biochemical evidence for a conserved interaction between bacterial transferrin binding protein A and transferrin binding protein B; Fuller CA et al.; As an adaptation to the iron-restricted environment of the host, some bacterial pathogens possess iron acquisition pathways mediated by surface receptors that specifically bind transferrin from the host . The receptor is composed of two receptor proteins, TbpA and TbpB, which are both capable of binding to transferrin . Previous studies have demonstrated that affinity isolation of TbpB from Neisseria meningitidis or Haemophilus influenzae with immobilized human transferrin required the homologous TbpA, implicating a TbpA-TbpB interaction . In this study, we demonstrated that TbpA from either species can facilitate isolation of either TbpB, indicating that the TbpA-TbpB interaction is conserved within these species . Extension of these studies to veterinary pathogens in which a TbpA-Tf complex is used to affinity isolate heterologous TbpBs, demonstrated an interaction between the receptor proteins from N . meningitidis and Actinobacillus pleuropneumoniae . Further delineation of the TbpA-TbpB-transferrin interaction with recombinant chimeric N . meningitidis/A . pleuropneumoniae TbpBs has identified a region encoded by the first 1/4 of the tbpB gene which is involved in Tf binding .

J Antimicrob Chemother, 1998 Feb, 41(2), 253 - 8
Effects of cefixime or co-amoxiclav treatment on nasopharyngeal carriage of Streptococcus pneumoniae and Haemophilus influenzae in children with acute otitis media; Dabernat H et al.; A multicentre, open-label, randomized study was performed in 501 out-patients with acute otitis media, aged 6-36 months, to study the impact of treatment with either cefixime suspension 8 mg/kg/day bd or co-amoxiclav suspension 80 mg/kg/day tds for 10 days on nasopharyngeal carriage of Streptococcus pneumoniae and Haemophilus influenzae . Of 426 patients with nasopharyngeal cultures at entry to the trial, end of treatment and at follow-up visit (35 days after inclusion), significant changes in carriage of S . pneumoniae were observed . The proportion of penicillin-resistant S . pneumoniae was higher in the samples taken at the end of treatment and follow-up than in those taken at inclusion, while the total number of children with this microorganism was lower . The difference at the end of treatment was greater with co-amoxiclav than with cefixime . For H . influenzae the resistance rate remained steady while the number of children with this microorganism decreased . At follow-up there was no significant difference between the two groups in terms of nasopharyngeal positive culture for S . pneumoniae or H . influenzae . Despite these differences, successful clinical responses were similar at the end of treatment and at follow-up.

J Antimicrob Chemother, 1998 Feb, 41(2), 207 - 14
Sensitivity to sparfloxacin and other antibiotics, of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis strains isolated from adult patients with community-acquired lower respiratory tract infections: a European multicentre study . SPAR Study Group . Surveillance Programme of Antibiotic Resistance; Richard MP et al.; A survey of resistance to sparfloxacin was carried out in ten European countries, namely Slovakia, France, Germany, Great Britain, Hungary, the Republic of Ireland, Italy, The Netherlands, Portugal and Spain . Respiratory samples were collected from 4297 patients with lower respiratory tract infections and cultured for the presence of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis . Altogether 2101 strains were isolated and tested for their susceptibility to sparfloxacin, ciprofloxacin, erythromycin, tetracycline and penicillin G (S . pneumoniae) or amoxycillin (H . influenzae and M . catarrhalis) . Each country tested strains using methods commonly used in that country, and with breakpoints selected based on those used in that country . Penicillin resistance in pneumococci was seen in those countries in which it had been reported previously, namely Spain, France and Hungary . Only four strains of pneumococci were resistant to sparfloxacin (MIC > or = 2 mg/L), while ciprofloxacin-resistant strains were isolated more frequently, particularly in the Republic of Ireland and Hungary . Almost all of the strains of H . influenzae tested were resistant to erythromycin, (MIC50 > or = 4 mg/L), but all strains were highly sensitive to sparfloxacin (MIC90 < or = 0.06 mg/L) . The number of strains of H . influenzae producing beta-lactamase varied between countries, whereas most strains of M . catarrhalis produced beta-lactamase . In M . catarrhalis, erythromycin and tetracycline resistance was rare, but sensitivity to amoxycillin varied . Sparfloxacin was particularly active against H . influenzae and M . catarrhalis, and was the most active compound tested . Overall, the activity of sparfloxacin was greater than that of ciprofloxacin against all three pathogens, and resistance to it was rare.

Rev Neurol, 1998 Jan, 26(149), 34 - 7
{Epidemiology of Haemophilus influenzae type b, Neisseria meningitidis, and Streptococcus pneumoniae in children in the Valencia Community, Spain . Acute diseases study group}; Morant A et al.; OBJECTIVE: To asses the incidence of Haemophilus influenzae type b (Hib), Neisseria meningitidis and Streptococcus pneumoniae meningitis in children of de Valencian Community (VC), Spain, and to describe the microbiologic characteristics . MATERIAL AND METHODS: Prospective surveillance system with paediatrician and microbiologist participation of all public hospitals of the VC . Cases are children less than 15 with clinical meningitis and with isolation of Hib, N . meningitidis or S . pneumoniae from CSF of blood . RESULTS: From 1st December 1995 to 30th November 1996, 51 cases were declared, 33.3% were Hib, 49.0% N . meningitidis and 17.7% S . pneumoniae . The annual incidence of meningitis was 7.6 cases/100,000 < 15 years, 20.5/100,000 < 5 years and 56.2/100,000 < 1 year . 84.3% of the cases occurred in children younger than 5 . S . pneumoniae had the highest mortality . CONCLUSIONS: Hib is a frequent cause of meningitis in spite of that one third of children are vaccinated . 43% of the N . meningitidis isolated in meningitis are serogroup C . S . pneumoniae meningitis are more frequent in children less than one, and has a high mortality rate.

Medicina (B Aires), 1997, 57(2), 191 - 9
{Etiology of acute lower respiratory tract infections among children younger than 5 years old in Santa Fe}; Sequeira MD et al.; The etiology of acute lower respiratory tract infections (ARI) and nasopharyngeal bacterial carriage in children less than 5 years old living in Santa Fe city, Argentina, was studied . A total of 518 children were included in the study: 450 suffering from ARI and 68 asymptomatic children . Blood samples, pleural effusions and nasopharyngeal secretions (NS) were obtained from children for bacterial isolations . NS was also used for fluorescent antibody techniques, and serum samples were employed for detecting IgM anti Chlamydia trachomatis . A bacterial pathogen was isolated from blood in 6.2% (14/224) of the children with ARI . A total of 11 Streptococcus pneumoniae (five of them oxacillin resistant), two Haemophilus influenzae and one Staphylococcus aureus strains were isolated . The most frequently detected pathogen in the ARI group was respiratory syncytial virus (RSV) . It was found in 23.3% (105/450) of the children with ARI . Among children with risk of Chlamydia trachomatis infection, 24% presented high titters of specific IgM antibodies . Main bacteria carried in NS in the ARI group were H.influenzae (31.6%) and S . pneumoniae (23.4%) while viridans streptococci (26.5%), H.influenzae (23.5%) and Moraxella catarrhalis (22.1%) were more frequently isolated from controls . The most common pneumococcal types were 14 and 7 and the main type of H.influenzae was b biotype I . During the period of this study, the susceptibility of the pneumococcal isolates to oxacillin decreased from 60% to 50.8%, and the H.influenzae susceptibility to ampicillin fell from 92.3% to 79% . All the H.influenzae type b isolations were susceptible to ampicillin.

J Pediatr Orthop, 1998 Mar-Apr, 18(2), 262 - 7
Increasing prevalence of Kingella kingae in osteoarticular infections in young children; Lundy DW et al.; Sixty children younger than 3 years with culture-positive hematogenous septic arthritis and acute/subacute osteomyelitis treated between 1990 and 1995 were reviewed to identify the infecting organism . Gram-positive bacteria were identified in 47 (78.3%) patients, and gram-negative organisms were identified in 13 (21.7%) patients . Haemophilus influenzae was cultured in none of the cases of septic arthritis and in only one (1.6%) case of acute osteomyelitis . Kingella kingae was cultured in 10 (16.7%) cases, with all of these patients between the ages of 10.5 and 23.5 months . Routine immunization of infants against H . influenzae has caused a change in the historically reported bacteria of bone and joint infections in children younger than 3 years . Haemophilus influenzae has lost its predominance as the most commonly identified gram-negative pathogen, and in this study, has been replaced by K . kingae.

Drugs, 1998 Mar, 55(3), 347 - 66
Meningococcal vaccines . Current status and future possibilities; Peltola H; Meningococcal disease causes great emotion and anxiety in the families and caregivers of patients . Numbers of such patients are usually small in industrialised countries, unlike those in many regions--especially in subsahelian Africa . Vaccines have been tried for more than 80 years; at present there are available polysaccharide vaccines against groups A, C, Y and W135, and a protein-based vaccine against group B . A property common to all is their relative efficacy (75 to 100%) at school age and after, and an acceptably short persistence of antibodies . Small children pose the major challenge, in whom there is essentially evidence of clinical protection only against group A and C diseases . With vaccines against other serogroups protection is possible, but not yet proven in controlled clinical studies . The search is on for help from various modifications, including the conjugation technique, to transform the independent nature of polysaccharide response towards T cell dependence, as was done earlier in Haemophilus influenzae type b vaccines . First trials along this path are encouraging although, again, group B meningococci pose special problems . The next few years will probably see a new generation of meningococcal vaccines . Generally speaking, the incidence of meningococcal disease is too low to indicate vaccinations for the whole population, or even children, but some risk groups and epidemics are important exceptions . To date, bivalent group A + C or tetravalent group A + C + Y + W135 polysaccharides, or an outer membrane protein-based group B vaccine, are the products to be used when the indications, that may vary from country to country, are considered met . A strong herd immunity effect, demonstrated with group A and C vaccinations, facilitates extinction of an epidemic since large-scale vaccinations can be restricted only in the major risk groups, children and in various schools . Prompt intervention demands, however, a functioning mechanism which detects very early on a pending epidemic . Unfortunately, such a mechanism is often lacking in countries often hit by this deadly disease.

Curr Opin Pediatr, 1998 Feb, 10(1), 13 - 8
Bacterial meningitis and meningococcal infection; Booy R et al.; Now that invasive disease caused by Haemophilus influenzae type b has been largely controlled in the developed world, the principal bacterial pathogen causing meningitis in most countries is the meningococcus . Serogroup B and C strains predominate in most industrialised countries in contrast to the situation in sub-Saharan Africa, where serogroup A meningococcal disease is periodically a problem of massive epidemic proportions . This review focuses on developments in our understanding of the diagnosis and treatment of meningococcal disease, of host factors important in susceptibility to, and severity of, this infection, and on advances which may eventually lead to its prevention.

Infect Immun, 1998 Apr, 66(4), 1752 - 4
Aging and the immune response to the Haemophilus influenzae type b capsular polysaccharide: retention of the dominant idiotype and antibody function in the elderly; Lucas AH et al.; Anti-Haemophilus influenzae b polysaccharide (Hib PS) antibodies elicited in elderly subjects following conjugate vaccination expressed a light-chain variable-region (VL)-associated idiotype and had functional activities similar to those previously observed in children and younger adults . These findings indicate that advanced age is not accompanied by shifts in the major VL component of the Hib PS-specific repertoire or by diminution of the protective function of antibodies.

Infect Immun, 1998 Apr, 66(4), 1622 - 31
Copy number of pilus gene clusters in Haemophilus influenzae and variation in the hifE pilin gene; Read TD et al.; Brazilian purpuric fever (BPF)-associated Haemophilus influenzae biogroup aegyptius strain F3031 contains two identical copies of a five gene cluster (hifA to hifE) encoding pili similar to well-characterized Hif fimbriae of H . influenzae type b . HifE, the putative pilus tip adhesin of F3031, shares only 40% amino acid sequence similarity with the same molecule from type b strains, whereas the other four proteins have 75 to 95% identity . To determine whether pilus cluster duplication and the hifE(F3031) allele were special features of BPF-associated bacteria, we analyzed a collection of H . influenzae strains by PCR with hifA- and hifE-specific oligonucleotides, by Southern hybridization with a hifC gene probe, and by nucleotide sequencing . The presence of two pilus clusters was limited to some H . influenzae biogroup aegyptius strains . The hifE(F3031) allele was limited to H . influenzae biogroup aegyptius . Two strains contained one copy of hifE(F3031) and one copy of a variant hifE allele . We determined the nucleotide sequences of four hifE genes from H . influenzae biogroup aegyptius and H . influenzae capsule serotypes a and c . The predicted proteins produced by these genes demonstrated only 35 to 70% identity to the three published HifE proteins from nontypeable H . influenzae, serotype b, and BPF strains . The C-terminal third of the molecules implicated in chaperone binding was the most highly conserved region . Three conserved domains in the otherwise highly variable N-terminal putative receptor-binding region of HifE were similar to conserved portions in the N terminus of Neisseria pilus adhesin PilC . We concluded that two pilus clusters and hifE(F3031) were not specific for BPF-causing H . influenzae, and we also identified portions of HifE possibly involved in binding mammalian cell receptors.

Rev Neurol, 1997 Dec, 25(148), 1886 - 9
{Subdural effusion in type B Haemophilus influenzae meningitis . A study of 38 cases}; Fernandez-Jaen A et al.; OBJECTIVE: Studying clinical, laboratory and radiologic findings, as well as outcome, observed in patients with meningitis caused by Hib, and its relationship with subdural effusion . MATERIAL AND METHODS: Retrospective study of 38 meningitis caused by Hib . Patients were aged between 3 months and 5 years . Imaging was performed in 26 cases (68%): CT in 21 children (55%) and cranial sonography in 11 cases (29%) . EEG was made in 29 patients (76%) and auditory-evoked potentials in 13 (34%) . The mean follow-up period after discharge was 24 months . RESULTS: Sixty-six per cent were male and 34% female . Eight cases had subdural effusion . These patients showed higher white cell counts in blood and CSF, higher levels of proteins in CSF, and lower levels of glucose in the same medium . They also had seizures before or during hospitalization, with higher frequency than those without subdural effusion (50% vs 26%) as well as more prolonged fever (127 vs 73 hours) . No specific treatment was required in any case . CONCLUSIONS: Subdural effusion is one of the most frequent complications observed in meningitis . Patients frequently present more important clinical and laboratory alterations . This finding is not related with neurologic sequelae and they resolve spontaneously with time.

Antimicrob Agents Chemother, 1998 Feb, 42(2), 319 - 24
Antimicrobial susceptibility of Haemophilus influenzae in the respiratory tracts of patients with cystic fibrosis; Moller LV et al.; We analyzed the antimicrobial susceptibilities of Haemophilus influenzae isolates from 157 sputum specimens prospectively collected from 39 cystic fibrosis (CF) patients during a 2-year study . These isolates were characterized by random amplified polymorphic DNA analysis and major outer membrane protein (MOMP) analysis to identify H . influenzae strains and MOMP variants and to assess their persistence in the respiratory tract . Among the 247 H . influenzae isolates, 16 (6.5%) produced beta-lactamase . The 231 beta-lactamase-negative isolates represented 85 H . influenzae strains, 61 MOMP variants derived from 27 of these strains, and 85 persistent isolates identical to strains or MOMP variants . All beta-lactamase-negative isolates were tested for susceptibility to ampicillin, amoxicillin-clavulanic acid, cefuroxime, cefotaxime, cefaclor, imipenem, tetracycline, and trimethoprim-sulfamethoxazole by disk diffusion testing . Eleven (13%) H . influenzae strains, 18 (30%) MOMP variants, and 30 (35%) persistent isolates were resistant to one or more of the antibiotics tested . Antimicrobial susceptibility was decreased among MOMP variants and persistent isolates compared to nonpersistent H . influenzae strains, and changes in susceptibility occurred irrespective of MOMP variation . We conclude that the decreased antimicrobial susceptibility of H . influenzae during persistence contributes to the poor eradication of H . influenzae from the respiratory tracts of CF patients.

Enferm Infecc Microbiol Clin, 1997 Nov, 15(9), 462 - 7
{Acute epiglottitis caused by Haemophilus influenzae type b in children: presentation of 21 cases}; del Carmen Otero M et al.; BACKGROUND: The aim of this paper is to review the epidemiological, clinical and therapeutical characteristics of Haemophilus influenzae type b (Hib) epiglottitis in children at a time when an efficient and safe vaccination is available . METHODS: The clinical histories of 21 children admitted to Children's Hospital La Fe (1971-1996) with a clinical diagnosis of epiglottitis and isolation of the microorganism in blood cultures (20 cases) and surface culture of the epiglotis (one case) are reviewed . RESULTS: The annual average was 4/100,000 children under 5 years of age . Evolution prior to diagnosis was > 12 hours in 52.4% of the cases . More males were affected (52.4% vs 47.6%) . All the children except one (95.2%) were under 5 years of age; 81% were under 3 years of age and 1 child was 6 years and 8 months old . Respiratory distress (100%) and fever > or = 38 degrees C (85.7%) were the most common clinical manifestations . General health was affected in 71.4% of the cases and 66.7% had leucocytosis on admission . The clinical diagnosis was confirmed by direct visualization of the epiglotis in 76.1% of the cases . Hib was isolated in blood culture in 20 cases (95.2%) . The strains produced beta-lactamases and were ampicillin-resistant in 57.1% . 19 children (90.5%) required endotracheal intubation . Initial empiric antibiotic therapy was third generation cephalosporins (cefotaxime or ceftriaxone) alone or combined with ampicillin . One child died (4.8%) . CONCLUSIONS: Pediatricians must still be aware of this serious infection in order to diagnosis and treat it as early as possible.

Can J Microbiol, 1998 Jan, 44(1), 91 - 4
Functional characterization of the Haemophilus influenzae 4.5S RNA; Jenkins GS et al.; The putative 4.5S RNA of Haemophilus influenzae was identified in the genome by computer analysis, amplified by the polymerase chain reaction, and cloned . We have determined that this putative 4.5S RNA will complement an Escherichia coli strain conditionally defective in 4.5S RNA production . The predicted secondary structures of the molecules were quite similar, but Northern analysis showed that the H . influenzae RNA was slightly larger than the E . coli RNA . The H . influenzae gene encoding this RNA is the functional homolog of the ffs gene in E . coli.

Clin Ther, 1998 Jan-Feb, 20(1), 141 - 55
Efficacy and tolerability of twice-daily ciprofloxacin 750 mg in the treatment of patients with acute exacerbations of chronic bronchitis and pneumonia; Pryka R et al.; In a review of the US Bayer ciprofloxacin (CIP) database, an analysis was undertaken to summarize the effectiveness and tolerability of CIP 750 mg BID in the treatment of patients with acute exacerbations of chronic bronchitis (AECB) and pneumonia . In five controlled studies, comparator (COMP) agents included ampicillin, intravenous cefuroxime/cefaclor, and other unspecified agents . Primary efficacy end points were clinical success (resolution plus improvement) and bacteriologic eradication at the end of therapy . The incidence of adverse events for CIP 750 mg BID was compared with that for COMP and with that in the CIP 500-mg-BID AECB and pneumonia clinical trials database . In five uncontrolled studies, 443 patients received CIP 750 mg BID; in 5 controlled trials comprising 344 patients, 169 received CIP 750 mg BID and 175 received COMP . Clinical success for CIP was 93% (368/396) and 99% (160/162), respectively, in the uncontrolled and controlled studies versus 98% (156/160) for COMP agents . Corresponding bacteriologic eradication rates for CIP 750-mg-BID-treated patients were 77% (273/356) and 95% (122/128), respectively, and 77% (96/125) for COMP agents . Overall bacteriologic eradication by organism for CIP 750 mg BID included Streptococcus pneumoniae 96% (51/53), Haemophilus influenzae 98% (92/94), Haemophilus parainfluenzae 100% (56/56), Moraxella catarrhalis 100% (14/14; 13 of 14 organisms were isolated in patients with AECB), and Pseudomonas aeruginosa 66% (135/204) . Drug-related adverse events were reported in 113 (26%) CIP 750-mg-BID-treated patients in uncontrolled trials and in 62 (37%) CIP 750-mg-BID- and 61 (35%) COMP-treated patients in controlled trials . In the combined data from the CIP 750-mg-BID uncontrolled and controlled trials, adverse events occurred with similar frequency compared with COMP except for nausea (CIP 10%, COMP 7%) and diarrhea (CIP 3%, COMP 13%) . In conclusion, CIP 750 mg BID provided excellent clinical success rates in the treatment of patients with AECB and pneumonia . CIP 750 mg BID was well tolerated compared with the COMP agents administered.

Clin Diagn Lab Immunol, 1998 Mar, 5(2), 219 - 24
Blinded multiplex PCR analyses of middle ear and nasopharyngeal fluids from chinchilla models of single- and mixed-pathogen-induced otitis media; Bakaletz LO et al.; Multiplex PCR analyses for both bacterial and viral pathogens were conducted in a blinded manner on 33 archival specimens, of known culture status, procured from chinchilla models of both single- and mixed-pathogen-induced otitis media and from a pediatric patient . These specimens had been maintained at -70 degrees C for up to 6 years . Experimental specimens evaluated included middle-ear effusions, nasopharyngeal lavage fluids and middle-ear lavage fluids from animals which were immunologically naive, sham-immunized or actively immunized with nontypeable Haemophilus influenzae antigens . Sampling times used ranged from the day of bacterial or viral challenge to 42 days after challenge . Initial PCR analyses of the 33 specimens matched the traditional culture data in 24 instances (73%), correctly identifying nontypeable H . influenzae, Moraxella catarrhalis, Streptococcus pneumoniae, or adenovirus as the causative agent . A PCR-positive signal for the microbe(s) inoculated was also obtained in four animal model specimens (12%) which were culture negative . One of two culture-negative human effusions was also PCR positive . Thus, overall, results obtained by blinded PCR were 85% concordant with traditional culture methods or correctly indicated the specific pathogen introduced in four specimens that were sterile . In no instance was a false-positive signal obtained for any of the five etiologic agents being evaluated . We conclude that the multiplex PCR analyses are rapid and accurate methodologies when they are used to retrospectively evaluate diverse archival specimens of limited volume from experimental models of otitis media.

ORL J Otorhinolaryngol Relat Spec, 1998 Jan-Feb, 60(1), 35 - 41
Haemophilus influenzae and Streptococcus pyogenes group A challenge induce a Th1 type of cytokine response in cells obtained from tonsillar hypertrophy and recurrent tonsillitis; Agren K et al.; We have previously shown that tonsil tissue both from children with tonsillar hypertrophy and recurrent tonsillitis is colonized and invaded by Haemophilus influenzae and Streptococcus pyogenes group A . In order to evaluate if these bacteria are involved in the immunopathogenesis of these two conditions, tonsillar cells from both groups were stimulated in vitro with intact, heat-inactivated H . influenzae or S . pyogenes A . The immunoreactivity was evaluated by assessing the induction of cytokine production (IL-1 alpha, IL-1 beta, TNF-alpha, IL-6, IL-8, IL-2, IFN-gamma, IL-4, TNF-beta and IL-10), which was detected at the single-cell level . All cytokines studied except IL-4 were induced in both groups after stimulation with H . influenzae or S . pyogenes A . The dominating cytokines were IL-1 beta, IFN-gamma and TNF-beta . No major differences in the cytokine pattern or number of cytokine-producing cells were noticed between the two patient cohorts after H . influenzae stimulation . Activation by S . pyogenes A bacteria gave rise to higher frequencies of IFN-gamma- and TNF-beta-synthesizing cells in the recurrent tonsillitis group . The incidence of CD4-, CD8-positive T cells and CD40-positive B cells was comparable between the two groups while the MAC-387-positive macrophages were significantly higher in the recurrent tonsillitis groups . In conclusion, a Th1 type of cytokine response was found in both groups following stimulation with H . influenzae or S . pyogenes A.

Antimicrob Agents Chemother, 1998 Mar, 42(3), 589 - 95
Prevalence of antimicrobial-resistant pathogens in middle ear fluid: multinational study of 917 children with acute otitis media; Jacobs MR et al.; The management of acute otitis media is complicated by the emergence of resistance to beta-lactam and other antibiotics among common pathogens . We conducted a large, international study of infants and children with acute otitis media to identify pathogens and susceptibility patterns . During the winter of 1994 to 1995, middle ear fluid samples were collected from 917 patients with acute otitis media in Bulgaria, the Czech Republic, Hungary, Romania, Slovakia, Israel, and the United States . A single reference laboratory performed in vitro susceptibility testing . Pathogens were isolated from 62% of the patients . For Streptococcus pneumoniae (30% of the patients), untypeable Haemophilus influenzae (17%), and Moraxella catarrhalis (4%), there was significant variation among geographic regions (P < 0.001) . The composite susceptibilities of these three organisms to amoxicillin ranged from 62% in the United States to 89% in Eastern and Central Europe; the corresponding susceptibilities to amoxicillin-clavulanate ranged from 90% in Israel to 95% in Eastern and Central Europe . beta-Lactamase was produced by 31 and 100% of the isolates of H . influenzae and M . catarrhalis, respectively . More isolates of S . pneumoniae were susceptible to amoxicillin (90%) or amoxicillin-clavulanate (90%) than to penicillin (70%; P = 0.002) . The prevalence of resistant S . pneumoniae was highest in patients less than 12 months of age . S . pneumoniae, H . influenzae, and M . catarrhalis remain the most important bacterial pathogens in patients with acute otitis media; however, their prevalence is variable and resistance patterns are changing.

Am J Respir Crit Care Med, 1998 Mar, 157(3 Pt 1), 950 - 6
Haemophilus influenzae in lung explants of patients with end-stage pulmonary disease; Moller LV et al.; In order to determine the presence and distribution of Haemophilus influenzae in lung tissue sections, we obtained lung explants from 49 lung transplant recipients with cystic fibrosis (CF) (n = 16), chronic obstructive pulmonary disease (COPD) including emphysema (n = 16), bronchiectasis (n = 5), pulmonary hypertension (n = 9), Langerhans cell histiocytosis (n = 1), and idiopathic pulmonary fibrosis (n = 2) . Analysis was done by selective culturing, immunoperoxidase (IP) staining, and by polymerase chain reaction (PCR) . H . influenzae was cultured from specimens of the lung explants from one CF and one COPD patient . IP staining of tissue sections was positive in 24 patients (10 CF patients, eight COPD patients, two bronchiectasis patients, and four patients with noninfectious pulmonary diseases) . IP-positive tissue sections were PCR-positive, and IP-negative sections were PCR-negative . H . influenzae was more frequently detected in tissue sections of lung explants from CF and COPD patients than from patients with bronchiectasis or noninfectious pulmonary diseases . H . influenzae was diffusely present in the epithelium, the submucosa of the bronchi, the bronchioles, the interstitium, and the alveolar epithelium . H . influenzae was localized extracellularly alone and in bacterial clusters, and was also associated with macrophages in CF patients . The results of this study demonstrate that H . influenzae is often present in the lungs of patients with end-stage pulmonary disease, especially CF and COPD patients . H . influenzae is diffusely present in the respiratory epithelium and subepithelial layers of the lungs of these patients.

J Exp Med, 1998 Feb 16, 187(4), 631 - 40
Phosphorylcholine on the lipopolysaccharide of Haemophilus influenzae contributes to persistence in the respiratory tract and sensitivity to serum killing mediated by C-reactive protein; Weiser JN et al.; Haemophilus influenzae undergoes phase variation in expression of the phosphorylcholine (ChoP) epitope, a structure present on several invasive pathogens residing in the human respiratory tract . In this study, structural analysis comparing organisms with and without this epitope confirmed that variants differ in the presence of ChoP on the cell surface-exposed outer core of the lipopolysaccharide . During nasopharyngeal carriage in infant rats, there was a gradual selection for H . influenzae variants that express ChoP . In addition, genotypic analysis of the molecular switch that controls phase variation predicted that the ChoP+ phenotype was predominant in H . influenzae in human respiratory tract secretions . However, ChoP+ variants of nontypable H . influenzae were more sensitive to the bactericidal activity of human serum unrelated to the presence of naturally acquired antibody to ChoP . Serum bactericidal activity required the binding of C-reactive protein (CRP) with subsequent activation of complement through the classical pathway . Results of this study suggested that the ability of H . influenzae to vary expression of this unusual bacterial structure may correlate with its ability both to persist on the mucosal surface (ChoP+ phenotype) and to cause invasive infection by evading innate immunity mediated by CRP (ChoP- phenotype).

Microb Pathog, 1997 Dec, 23(6), 327 - 33
Effect of Haemophilus somnus on nitric oxide production and chemiluminescence response of bovine blood monocytes and alveolar macrophages; Gomis SM et al.; Haemophilus somnus is able to survive and multiply in bovine blood monocytes (BBM) and alveolar macrophages (BAM), but the mechanisms used by H . somnus to evade killing mechanisms of bovine mononuclear phagocytes are not completely understood . To study the bactericidal ability of bovine mononuclear phagocytes following interaction with H . somnus, in vitro assay systems were developed to detect the luminol-dependent chemiluminescence response (LDCL) and nitric oxide (NO) production of BBM and BAM . Live logarithmically growing or stationary phase H . somnus inhibited the LDCL of BBM and BAM costimulated with opsonized Staphylococcus aureus . Inhibition of the LDCL response of BBM and BAM was not mediated by live H . somnus opsonized with hyperimmune serum, or by killed bacteria . H . somnus stimulated both BBM and BAM to produce NO at levels comparable with Escherichia coli lipopolysaccharide . While NO was being produced, viable H . somnus could still be isolated from the cell cultures . The ability of H . somnus to inhibit LDCL of both BBM and BAM, and resistance to NO killing may be an important mechanism that contributes to survival of the organism following ingestion by bovine mononuclear phagocytes .

Chest, 1998 Mar, 113(3 Suppl), 205S - 210S
How do we achieve cost-effective options in lower respiratory tract infection therapy?
Grossman RF.
Acute bronchitis and acute exacerbations of chronic bronchitis, common illnesses encountered by general and family physicians, account for approximately 14 million physician visits per year . The pattern of antibiotic prescribing for these infections varies from country to country, but there is no clear rationale for these antimicrobial choices . A recent meta-analysis of all randomized, placebo-controlled trials of patients treated with antibiotics for acute exacerbations of chronic bronchitis concluded that a small but statistically significant improvement could be expected in antibiotic-treated patients . Haemophilus influenzae is the most commonly isolated organism from sputum in patients with acute exacerbations of chronic obstructive lung disease but other Haemophilus species, Streptococcus pneumoniae, and Moraxella catarrhalis may also be found . High-risk patients can be defined as being elderly, with significant impairment of lung function, having poor performance status with other comorbid conditions, having frequent exacerbations, and often requiring oral corticosteroid medication . Well-defined clinical trials measure efficacy of a drug but not the effectiveness in a real world situation . Future studies of new antimicrobials should examine their efficacy in patients with an increased risk of true bacterial infection.

Acta Biochim Pol, 1997, 44(3), 491 - 504
Why a "benign" mutation kills enzyme activity . Structure-based analysis of the A176V mutant of Saccharomyces cerevisiae L-asparaginase I; Bonthron DT et al.; A conservative and apparently harmless A176V mutation in intracellular S . cerevisiae L-asparaginase (ScerAI) completely abolishes the enzyme activity . Sequence and structural comparisons with type II bacterial L-asparaginases show that the mutated residue is in a very conservative region and plays a vital role in the cohesion of functional tetramers of these enzymes through participation in side-chain...main-chain (Ser) Oy...O (Ala) hydrogen bonds across the tetramer interface . The fact that bacterial L-asparaginases of type I show less conservation in this region suggests that they may have different quaternary structure while adopting the subunit fold and intimate dimer architecture of type II enzymes . A comparison of all available sequences of microbial L-asparaginases confirms that separate intra- and extra-cellular enzymes evolved in prokaryotes and eukaryotes independently . However, an analysis of the available complete genome sequences reveals a surprising fact that Haemophilus influenzae possesses only a type II asparaginase while the archaebacterium Methanococcus jannaschii has a type I gene, but not a type II.

Gene, 1998 Jan 30, 207(2), 251 - 7
The sodA gene of Haemophilus ducreyi encodes a hydrogen peroxide-inhibitable superoxide dismutase; San Mateo LR et al.; Haemophilus ducreyi is the etiologic agent of the sexually transmitted disease chancroid, an ulcerative condition implicated in increased HIV transmission . There is increasing evidence for the roles of oxidative stress proteins including superoxide dismutase enzymes in the survival and persistence of pathogenic organisms within the host . The sodA gene of Haemophilus ducreyi was isolated from a genomic plasmid library on the basis of its ability to rescue the hydrogen peroxide hypersensitivity of an Escherichia coli sodA sodB strain . The H . ducreyi SodA protein also complemented the aerobic growth defect of the E . coli sodA sodB strain in minimal medium . The deduced amino-acid sequence of the H . ducreyi sodA gene product is 74 and 70% identical to the Mn-SODs of Haemophilus influenzae and E . coli, respectively . However, unlike Mn-SODs, the H ducreyi SodA protein was inhibited by hydrogen peroxide in native gels stained for SOD activity.

J Otolaryngol, 1998 Feb, 27(1), 10 - 6
Ciprofloxacin versus cefuroxime axetil in the treatment of adult patients with acute bacterial sinusitis; Klein GL et al.; OBJECTIVE: This study compared the use and efficacy of ciprofloxacin to cefuroxime axetil for adult patients with acute bacterial sinusitis . METHOD: We conducted a prospective, randomized, double-blind pilot study of oral ciprofloxacin (500 mg twice daily) versus cefuroxime axetil (250 mg twice daily) for 2 to 3 weeks in the treatment of adult patients with a clinical diagnosis of acute bacterial maxillary sinus infections or acute exacerbation of chronic bacterial sinusitis . Patients with microbiologically and radiologically confirmed sinusitis infection composed the efficacy population . RESULTS: Of the 83 patients enrolled, 13 of 42 (31%) ciprofloxacin- and 19 of 41 (46%) cefuroxime axetil-treated patients had a respiratory pathogen isolated from a sinus aspiration . The most frequent pretherapy isolated included Haemophilus influenzae (11), streptococcus species (20), staphylococcus species (7), Proteus mirabilis (3), and Neisseria sicca (3) . At the end of therapy, clinical resolution or improvement in efficacy-valid patients was achieved in 12 (100%) ciprofloxacin-treated patients and in 14 (74%) cefuroxime axetil recipients . The five (26%) cefuroxime axetil clinical failures were due to development of superinfection . Bacteriologic eradication occurred in 12 (100%) and 14 (100%) ciprofloxacin and cefuroxime axetil patients, respectively . Similar clinical and bacteriologic response rates were observed at the 2- to 4-week follow-up . Among 83 intent-to-treat patients, 19 (45%) ciprofloxacin and 14 (34%) cefuroxime axetil patients had drug-related adverse events . The most common adverse event in both treatment groups was gastrointestinal . CONCLUSION: This pilot study suggests that ciprofloxacin is efficacious in the management of acute bacterial sinusitis.

J Antimicrob Chemother, 1997 May, 39 Suppl A, 69 - 73
Activity of quinupristin/dalfopristin and its components against Haemophilus influenzae; Jorgensen JH et al.; Quinupristin/dalfopristin is an injectable streptogramin antibiotic that is constituted in a 30:70 (w/w) ratio of the two components . Quinupristin and dalfopristin are thought to act synergically by binding to two separate sites on the bacterial 50S ribosomal subunit . The in-vitro activities of the two components separately and together in different ratios were determined for a collection of 100 Haemophilus influenzae strains representing various antimicrobial resistance phenotypes . The NCCLS microdilution susceptibility testing procedure incorporating Haemophilus test medium (HTM) broth was used to determine MICs of quinupristin, dalfopristin and seven other antimicrobial agents . The MIC50 and MIC90 values were 4 and 8, 4 and 16, and 64 and 128 mg/L for quinupristin/dalfopristin (30:70), dalfopristin and quinupristin, respectively . MICs of quinupristin and dalfopristin were also determined in Mueller-Hinton lysed horse blood broth and by HTM agar dilution testing . Compared with HTM broth-derived results, the MICs of quinupristin/dalfopristin and its components were the same or one dilution higher in lysed horse blood and HTM agar incubated in air, and were equivalent or one dilution lower in HTM agar incubated in a CO2 atmosphere . The MICs of quinupristin and dalfopristin separately or together were directly proportional to erythromycin MICs, but were otherwise unaffected by any of the resistance mechanisms represented in these strains . MICs of quinupristin and dalfopristin combined in ratios of 10:90, 70:30 and 90:10 did not differ significantly from those of the 30:70 ratio . Thus, unlike the synergic activity noted against Gram-positive bacteria, the activity of quinupristin/dalfopristin against H . influenzae appears to be due almost entirely to the dalfopristin component of the combination.

J Clin Microbiol, 1998 Mar, 36(3), 818 - 9
Haemophilus parainfluenzae liver abscess after successful liver transplantation; Friedl J et al.; Haemophilus parainfluenzae was isolated from a bile specimen and from an aspirate of a liver abscess in a 58-year-old liver-transplanted woman that was indicative of an invasion of the graft by an ascending route . Drug therapy, immunosuppression, rejection therapy, and Roux-en-Y choledochojejunostomy may have contributed to the septic course . Interdisciplinary cooperation was instrumental in diagnosis and successful management in this case.

Bull Soc Pathol Exot, 1997, 90(5), 339 - 41
{Infection profile in sickle cell anemia}; Diop S et al.; Our objectives were to determine aetiology and localisation of infection in sickle cell anaemia patients . The origin of fever was malaria in 47% of cases, 50% of bacterial infections and 3% of viral infections . Respiratory infections were concerned in 61% of cases, versus 24% of osteomyelitis . Salmonella were found in 37%, Haemophilus (16%), Staphylococcus (14%), Streptococcus (10%) and Pneumococcus (9%) . We found more bacterial infection in anaemic forms (SS and SFA2) and more bacterial infection anemic forms (SC, SAFA2) . In view of these findings, we preconize malaria prophylaxis and vaccination against Salmonella, Haemophilus and Pneumococcus in sickle cell anemia patients.

Electrophoresis, 1997 Dec, 18(15), 2968 - 77
Large-scale identification of proteins of Haemophilus influenzae by amino acid composition analysis; Fountoulakis M et al.; Two-dimensional protein maps of microorganisms are useful tools for elucidation and detection of target proteins, a process essential in the development of new pharmaceutical products . We applied amino acid composition analysis, following separation by two-dimensional gel electrophoresis, for large-scale identification of proteins of Haemophilus influenzae . H . influenzae is a bacterium of pharmaceutical interest of which the entire genome, comprising approximately 1700 open reading frames, has been sequenced . For amino acid analysis, we used both precolumn derivatization of amino acids followed by reversed-phase chromatography of the derivatized residues and post-column derivatization of the residues previously separated on an ion exchanger . The composition analyses derived from both methods allowed the identification of 110 protein spots . The proteins were identified using the AACompldent software on the ExPASy server accessible via the World Wide Web with a success rate of 52% . In some cases, introduction of the analysis data of 12 residues was sufficient for a correct identification . Proteins which contained an unusually high percentage of one residue could be unambiguously identified . Amino acid composition analysis proved to be an error-robust, efficient method for protein identification . The method can be practically established in every biochemical laboratory and, complementary to mass spectrometry, represents an important analytical tool for the mapping of the proteomes of organisms of interest.

J Pediatr Ophthalmol Strabismus, 1998 Jan-Feb, 35(1), 38 - 40
Lacrimal-duct-probing-induced bacteremia: should children with congenital heart defects receive antibiotic prophylaxis?
Eippert GA, Burnstine RA, Bates JH.
PURPOSE: To determine the incidence of bacteremia following lacrimal-duct probing in children, and to evaluate the potential need for antibiotic prophylaxis in children who are at an increased risk of infectious endocarditis (IE) . METHODS: In a prospective study beginning in October 1994, 40 consecutive children requiring lacrimal-duct probing performed by a single pediatric ophthalmologist received preoperative lacrimal and blood cultures followed by postoperative probe-induced transient bacteremia that was defined as a negative preoperative blood culture, followed by a positive postoperative blood culture of the same bacteria identified in the positive lacrimal culture . RESULTS: The overall infection rate as described above was 7 of 40 children (17.5%) with a 95% confidence interval of 7.3% to 32.8% . Of these, four children had positive postoperative cultures for Haemophilus influenzae (10.0%) and 3 were positive for Streptococcus pneumoniae (7.5%) One child had a negative preoperative blood and lacrimal culture with a postoperative blood culture positive for Streptococcus viridans . CONCLUSIONS: This study shows a significant incidence of lacrimal-probe-induced bacteremia with organisms that have been documented as etiological agents for IE in children . IE, although less common in children, remains a serious, potentially life threatening infection with high mortality . Although lacrimal-duct probing has never been clearly associated with documented endocarditis, it is the authors' recommendation that it be prudent for patients who are at known high risk for endocarditis to receive SBE prophylaxis considering the low cost/benefit ratio.

Proc Natl Acad Sci U S A, 1998 Feb 17, 95(4), 1647 - 52
Abundant microsatellite polymorphism in Saccharomyces cerevisiae, and the different distributions of microsatellites in eight prokaryotes and S . cerevisiae, result from strong mutation pressures and a variety of selective forces; Field D et al.; We examined the distributions of short tandemly repeated DNAs (microsatellites) in nine complete microbial genomes (Saccharomyces cerevisiae, Archaeoglobus fulgidus, Escherichia coli, Haemophilus influenzae, Helicobacter pylori, Methanococcus jannaschii, Mycoplasma pneumoniae, M . genitalium, and Synechocystis PCC6803.) These repeats contribute differently to the global features of these genomes, and we explore the evolutionary implications of these differences by empirical examination of length polymorphisms at 20 long triplet-repeats repeats in S . cerevisiae, and by comparison of observed and expected repeat distributions . All of a sample of 20 microsatellites found in S . cerevisiae are highly polymorphic in length, suggesting that mutation pressure overcomes overall selection for small genome size that will tend to shorten or eliminate unnecessary DNA . By comparison, prokaryotes have fewer long repeats than expected, except for a few statistically improbable repeats that appear to function in gene regulation . Finally, we find that in all these genomes there is an excess of repeats shorter than those traditionally considered to be microsatellites . This finding suggests that even in prokaryotes these repeats are being generated by mutational pressures . These results have important potential implications for understanding genome stability and evolution in these microbial species.

Pharmacol Res, 1997 Dec, 36(6), 481 - 4
Bacterial adherence in smokers and non-smokers; Piatti G et al.; Bacterial adherence is thought to be a first important step in the pathogenesis of infection . It is now recognized that bacteria bind to and colonize mucosal surfaces in a highly selective manner via a lock- and key mechanism with complementary receptors on the mucosal surfaces of the host . We studied adherence to buccal cells of a panel of potential respiratory pathogens as Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae and Pseudomonas aeruginosa in 18 smokers and 18 healthy non-smokers . Our results show an increased pneumococcal adherence in smokers compared to that of non-smokers and this may explain the role of smoking as a risk factor in the susceptibility to bacterial pneumonia . The other bacterial species tested do not differ in their adhesion values and probably require previous damage of the mucosa before adhesion 1997 The Italian Pharmacological Society.

Microb Pathog, 1997 Nov, 23(5), 273 - 84
Characterization of the Pasteurella haemolytica transferrin receptor genes and the recombinant receptor proteins; Ogunnariwo JA et al.; The tbpA and tbpB genes encoding the transferrin receptor proteins, TbpA and TbpB, from Pasteurella haemolytica A1 were cloned, sequenced and expressed in Escherichia coli . The genes were organized in a putative operon arrangement of tbpB- tbpA . The tbpB gene was preceded by putative promoter and regulatory sequences, and followed by a 96 base pair intergenic sequence in which no promoter regions were found, suggesting that the two genes are coordinately transcribed . The deduced amino acid sequences of the TbpA and TbpB proteins had regions of homology with the corresponding Neisseria meningitidis, N . gonorrhoeae, Haemophilus influenzae and Actinobacillus pleuropneumoniae Tbp and Lbp proteins . The intact tbpB gene was expressed in a T7 expression system and the resulting recombinant TbpB protein retained the functional bovine transferrin binding characteristics . The availability of the recombinant TbpB enabled us to demonstrate its specificity for ruminant transferrins, its ability to bind both the C-and N-terminal lobes of bovine transferrin and its preference for the iron-loaded form of this protein . Several attempts at expressing the cloned tbpA gene were unsuccessful, suggesting that the product of the gene may be toxic to E . coli .

Infection, 1998 Jan-Feb, 26(1), 68 - 75
Randomized comparison of once-daily ceftibuten and twice-daily clarithromycin in the treatment of acute exacerbation of chronic bronchitis; Ziering W et al.; In an evaluator-blind, parallel-group, multicenter study, the efficacy and tolerability of ceftibuten 400 mg capsules once daily were compared with clarithromycin 500 mg twice daily for 7-14 days in the treatment of 309 patients with acute exacerbation of chronic bronchitis (AECB) . Clinical (n = 262) and microbiological (n = 71) assessments were conducted before treatment, during days 4-6 of treatment, and at 0-6 and 7-21 days after treatment . Clinical efficacy success rates (cure/improvement) at the end of treatment (0-6 days) were 91.0% for ceftibuten and 93.0% for clarithromycin . In the intent-to-treat population, the overall clinical assessment showed a success rate of 77.6% (121/156) in the ceftibuten group and 78.4% (120/153) in the clarithromycin group (95% confidence interval, -10.8 to +9.0%) . One patient in each of the ceftibuten and clarithromycin groups had a microbiological relapse and became a treatment failure . The overall success rate was 84.3% for ceftibuten and 86.7% for clarithromycin (C.I . -11.7%, +6.9) . Overall eradication of the target pathogens (Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae) was 84.8% for ceftibuten and 89.5% for clarithromycin . Eradication rates for ceftibuten at 0-6 days post treatment were 95.2% (H . influenzae), 87.5% (M . catarrhalis), and 100% (S . pneumoniae), compared with 85.7%, 100% and 100%, respectively, for clarithromycin . Significantly fewer patients in the ceftibuten group experienced treatment-related adverse events than in the clarithromycin group (5.3 vs 21.9%; p < 0.001) . This difference was due to a large number of patients in the clarithromycin group reporting taste perversion (12.6%) or gastrointestinal adverse events (9.9%) . Given its tolerability and efficacy profiles, and the advantage of once-daily administration, ceftibuten provides a rational alternative for the treatment of AECB.

An Esp Pediatr, 1997 Sep, 47(3), 263 - 8
{Review of the data on incidence of invasive disease and meningitis caused by Haemophilus influenzae in children under 5 years of age in Spain}; Guallar-Castillon P et al.; OBJECTIVE: The object of this study was to summarize the available information on the incidence of invasive disease and meningitis caused by Haemophilus influenzae in Spain from 1985 to 1995, evaluating potential geographic differences . METHODS: Systematic and exhaustive literature searches of computerized databases, manual review of obtained references, revision of relevant Spanish journals and proceedings of congresses and direct consultation with experts were carried out . Yearly incidence rates were estimated per 100,000 children less than 5 years of age, by year and geographical region . RESULTS: We encountered 16 studies, one covering the entire Spanish population and the rest limited to Catalonia, the Basque Country, the Valencian Community, Andalucia, Navarra or the Community of Madrid . In 1994, the incidence of invasive H . influenzae disease varied from 8.4 cases per 100,000 children under 5 years of age in Navarra to 26.3 cases per 100,000 children under 5 years of age in the Basque Country . CONCLUSIONS: Available data do not permit the nationwide estimation of the incidence rate, although it is possible to appreciate that important geographic differences exist . It is remarkable the lack of specific information for the majority of Spanish regions, as well as the lack of data to assess temporal trends.

Arch Dis Child, 1997 Dec, 77(6), 488 - 92
Antibody persistence and Haemophilus influenzae type b carriage after infant immunisation with PRP-T; Heath PT et al.; OBJECTIVES: To assess the persistence of serum Haemophilus influenzae type b antibodies and the prevalence of H influenzae type b carriage in a group of preschool age children previously vaccinated in infancy . DESIGN: Names were randomly selected from immunisation records . Families were visited on five occasions over a period of 12 months and throat swabs were taken from all family members present, with blood obtained from children at the first and last visits . RESULTS: One hundred and fifty three children at a median age of 3.6 years had a geometric mean titre (GMT) of 1.06 micrograms/ml (95% CI 0.80 to 1.38) . Eight per cent had an undetectable antibody concentration, received a booster dose of plain PRP vaccine, and responded with concentrations > 2 micrograms/ml . GMT at 4.5 years of age was 0.89 microgram/ml (0.69 to 1.16) . Twelve children who had been exposed to H influenzae had a GMT of 4.7 v 0.8 micrograms/ml for those without exposure . CONCLUSIONS: Accelerated immunisation against H influenzae without a second year booster results in persistence of satisfactory serum concentrations of antibody to 4.5 years of age . In those with undetectable antibody, immunological memory may still be present.

Ther Umsch, 1998 Jan, 55(1), 8 - 12
{Routine vaccination in childhood}; Desgrandchamps D; Some major modifications of the recommended Swiss vaccination schedule have been introduced in spring 1996: 1 . The vaccination against pertussis using the new acellular vaccines, which cause fewer side-effects . This allows the administration of a fourth and a fifth dose at the age of 2 and 4-7 years, respectively . 2 . The consideration of combination vaccines, which contain the vaccinations against diphtheria, tetanus and pertussis as well as the vaccine against invasive diseases caused by Haemophilus influenzae type b . 3 . In order to increase the individual vaccination protection, a second dose of MMR vaccine is recommended, preferably at the age of 4-7 years . A general recommendation for hepatitis B vaccination during adolescence may be expected in 1998.

Rinsho Byori, 1998 Jan, 46(1), 17 - 25
{Haemophilus parainfluenzae antigens in IgA nephropathy}; Suzuki S et al.; IgA nephropathy (IgAN), a common glomerular disease, is characterized by the presence of IgA deposits, predominantly in the glomerular mesangium, and by mesangial proliferative glomerulonephritis (GN) . Concerning its pathogenesis, several investigators suggest that the deposited IgA is an antibody to viral, bacterial, or dietary antigens . Thus the antibody is probably produced as part of the specific host immune response to various environmental antigens . Such reports strengthen the possibility of a relationship between mucosal immunity and the pathogenesis of IgAN . Nevertheless, attempts to isolate a specific IgA-circulating immune complex associated antigen in patients with IgAN have been unsuccessful . We have showed that such mucosal infections as pharyngitis are often associated with the acute onset of IgAN . Then IgAN is an immune complex disease that is caused by a poor mucosal immune response to environmental antigens to which the patient has been chronically exposed . We observed that Haemophilus parainfluenzae (HP) is more commonly isolated from the pharynx of patients with IgAN than from those with other diseases . We have also identified the glomerular deposition of outer membranes of HP antigens (OMHP) and an increased serum concentration of IgA antibodies against OMHP in patients with IgAN . Further studies will be necessary to determine whether the association of OMHP antigens in the glomeruli and IgA antibody against OMHP antigens in the sera of patients with IgAN can be confirmed in other parts of the world and whether this association is important in the pathogenesis of IgAN . Nevertheless, the demonstration of glomerular deposition of OMHP antigens and of IgA antibody against OMHP in sera indicates a potential new avenue of investigation into the elusive cause of IgAN.

Indian Pediatr, 1997 Sep, 34(9), 779 - 83
Immunogenicity study of Haemophilus influenzae type B conjugate vaccine in Indian infants; Phadke MA et al.; OBJECTIVE: To assess the immunogenicity in Indian infants to Haemophilus influenzae b oligosaccharide conjugate vaccine (HbOC) . DESIGN: Prospective multicenter study . SETTING: Pediatric Out Patient Department of general hospitals in Pune and Mumbai . SUBJECTS: 124 full term healthy infants brought for routine DPT/OPV immunization . METHODS: Infants were administered 3 doses of 0.5 ml of HbOC, on the same day as their DPT/OPV immunization, injected intramuscularly on the limb opposite to that where DPT vaccine was administered . Data on local reactions and general symptoms was collected for three days after every dose . The children had their blood collected for assay of anti PRP (polyribosil ribitol phosphate) antibody titers, along with the first injection and one month after the third injection . One hundred and three infants completed the study protocol with two blood collections . RESULTS: The initial geometric mean titers (GMT) of 0.124 mcg/ml rose by 37 times to 4.552 mcg/ml . Ninety eight children (95.1%) had a final titer of > or = 0.15 mcg/ml, the minimum level associated with protection, and 77 children (74.8%) had a final level of > or = 1.0 mcg/ml, a level associated with long term protection . CONCLUSION: HbOC is immunogenic in Indian infants when used as per the locally recommended DPT/OPV immunization schedule.

Pediatrics . 1998 Mar;101(3):E4.
Blood cultures in the evaluation of children with cellulitis; Sadow KB et al.; OBJECTIVE: To evaluate the yield of blood cultures obtained from immunocompetent children admitted for cellulitis in the post-Haemophilus influenzae type b (Hib) vaccine era and to determine whether these cultures are cost-effective . DESIGN: Retrospective case series . SETTING: Urban pediatric emergency department . Study Population . Patients 2 days to 22 years of age admitted with cellulitis from 1994 through 1995 . MEASUREMENTS AND RESULTS: Of 381 patients identified, 266 (70%) had blood cultures and 243 of these children were enrolled . Data recorded include demographics, immunization status, initial clinical appearance, antibiotic pretreatment, preexisting illness, location and precipitating cause of cellulitis, white blood cell count, and band-to-neutrophil ratio (BNR) . Blood cultures were categorized as positive, negative, or contaminant . Five cultures (2%) were positive, and 13 (5.4%) were contaminants . The positive blood cultures grew streptococcus and staphylococcus organisms, and none of the children were bacteremic with H influenzae . All patients with group A beta-hemolytic streptococcus had active varicella . The mean age was lower (26 vs 75 months) in those with a positive blood culture, and mean BNR was higher (0.32 vs 0.07) . Patient management did not change for bacteremic patients with uncomplicated cellulitis . All repeat cultures were negative . The cumulative charge for all blood cultures was $50 986 . CONCLUSIONS: Blood cultures are not cost-effective and are more frequently contaminated than positive in the evaluation of a patient with uncomplicated cellulitis . Since introduction of the H influenzae type b vaccine, the most common organisms are streptococci . Using a BNR = 0.20 as a threshold for sending blood cultures, we would have missed one positive culture, but would have avoided blood cultures in 213 patients (88%) with an estimated savings of $42 850.

Pediatrics . 1998 Mar;101(3):E3.
Interleukin-6, C-reactive protein, and abnormal cardiorespiratory responses to immunization in premature infants; Pourcyrous M et al.; OBJECTIVE: We report our experience with routine immunization of 89 premature infants in the neonatal intensive care unit because 1) a substantial number of them developed abnormal clinical signs, and 2) all but one of those who received diphtheria, tetanus, and whole-cell pertussis (DTwP) vaccine responded with elevations of interleukin-6 (IL-6) and C-reactive protein (CRP) concentrations that are otherwise characteristic of bacterial disease . METHODOLOGY: We hypothesized that the elevated IL-6 and CRP levels were solely a response to immunization and that treatment with antibiotics was not necessary . We performed this study in two consecutive parts . In part 1, we prospectively evaluated 79 consecutive premature infants who were immunized with DTwP, Haemophilus b conjugate vaccine, hepatitis B vaccine, and inactivated polio vaccine, (Hib, HBV, and IPV) . IL-6 and CRP were determined before immunization and every 12 hours on three occasions after immunization . In part 2, we studied an additional 10 infants who received acellular pertussis vaccine (DTaP) and who, 2 days later, received Hib, HBV, and IPV immunization simultaneously . We followed the same schedule of IL-6 and CRP determinations as in part 1 . RESULTS: In part 1, 24 infants (30%) developed abnormal cardiorespiratory signs within 24 hours after immunization . CRP and IL-6 values rose to abnormal levels after immunization in all but one infant; that infant was later shown to have a T-cell abnormality . In part 2, 3 infants had abnormal cardiorespiratory signs after simultaneous immunization with Hib, HBV, and IPV, but not after DTaP . IL-6 and CRP levels remained normal in all 10 infants . CONCLUSIONS: Part 1 demonstrates clearly the temporal relationship between IL-6 and CRP increments after DTwP, Hib, HBV, and IPV vaccines . In part 2 (DTaP was substituted for DTwP), there were no elevations of IL-6 or CRP, thus indicating that whole-cell pertussis component of DTwP was responsible for IL-6 and CRP elevations . Abnormal cardiorespiratory signs occurred frequently after immunizations in part 1, but they were unrelated to the magnitude of IL-6 and CRP elevations . The frequency of cardiorespiratory difficulty and its occasional severity suggest a need to monitor premature infants for approximately 48 hours after routine immunization.

Eur J Biochem, 1998 Jan 15, 251(1-2), 54 - 64
Overexpression in Escherichia coli of the rnf genes from Rhodobacter capsulatus--characterization of two membrane-bound iron-sulfur proteins; Jouanneau Y et al.; The rnf genes of Rhodobacter capsulatus, essential for nitrogen fixation, are thought to encode a system for electron transport to nitrogenase . In the present study, we have attempted to overexpress the rnf genes in Escherichia coli to investigate the molecular properties of the corresponding proteins . Corrections were made to the published DNA sequence of the rnf operon, resulting in the identification of two genes, rnfG and rnfH . The rnfABCDGEH operon thus comprises seven genes and shows similarities in gene arrangement and deduced protein sequences to homologous regions in the genomes of Haemophilus influenzae and E . coli . Four of the rnf gene products were found to be similar in sequence to components of an Na+-dependent NADH:ubiquinone oxidoreductase from Vibrio alginolyticus . Three of the rnf genes were successfully overexpressed in E . coli as His-tagged polypeptides, whereas the products of rnfA, rnfD and rnfE, predicted to be transmembrane proteins, could not be stably maintained in E . coli . The rnfB and rnfC gene products were isolated as two brown proteins with apparent molecular-mass values of 25 kDa and 55 kDa, respectively . RnfB was shown to contain one {2Fe-2S} cluster, based on absorption spectrophotometry, EPR spectroscopy and iron content . Recombinant RnfC contained at least one iron-sulfur cluster, most likely of the {4Fe-4S} type . Unambiguous identification of the prosthetic groups was, however, precluded by the extreme instability of this protein . In R . capsulatus, RnfB and RnfC were found by immunoblot analysis to be tightly bound to the membrane, despite their hydrophilic character . The RnfB and RnfC proteins were absent in mutant strains bearing insertions at various positions within the rnfABCDGEH operon, suggesting that their stability depends on the cosynthesis of the other rnf gene products . We observed that iron limitation during growth resulted in a decrease both in the cellular content of RnfB and in the level of transcription of the rnfABCDGEH operon, indicating that the expression of this operon is regulated as a function of iron availability.

Mol Biol Evol, 1998 Jan, 15(1), 17 - 27
The evolutionary relationships between the two bacteria Escherichia coli and Haemophilus influenzae and their putative last common ancestor; de Rosa R et al.; We have tried to approach the nature of the last common ancestor to Haemophilus influenzae and Escherichia coli and to determine how each bacterium could have diverged from this putative organism . The approach used was exhaustive analysis of the homologous proteins coded by genes present in these bacteria, using as criteria for sequence relatedness an alignment of at least 80 amino acid residues and a PAM distance (number of accepted point mutations per 100 residues separating two sequences) below 250 . Evolutionarily significant similarities were found between 1,345 H . influenzae proteins (85% of the total genome) and 3,058 E . coli . proteins (75% of the total genome), many of them belonging to families of various sizes (from 666 doublets to 35 large groups of more than 10 members) . Nearly all the genes found by this approach to be duplicated in both bacteria were already duplicated in their last common ancestor . This was deduced from (1) the comparison of the respective distributions of evolutionary distances between orthologs (genes separated only by speciation events) and paralogs (genes duplicated in the same genome) and (2) the analysis of the phylogenetic trees reconstructed for each family of paralogs containing at least two members belonging to each bacterium . The distributions of the different categories of homologs show a significant loss of paralogous genes in H . influenzae (reduction proportional to the genome size), of many sequences which are still present in one copy in E . coli, and of some entire gene families . Phylogenetic trees also confirmed this recent loss of paralogous genes in H . influenzae . Thus, the genome size of the last common ancestor of these two bacteria would have been close to that of present-day E . coli, and the evolution of H . influenzae toward a parasitic life led to an important decrease in its genome size by some mechanism of streamlining . During this recent evolution, the memory of the gene order present in the last common ancestor has been blurred, but a few short conserved chromosomal fragments can still be detected in present-day E . coli and H . influenzae.

Arch Pediatr, 1997 Nov, 4(11), 1119 - 24
{Virus-bacteria co-infections}; Floret D; Influenza is the best known model of bacterial-viral co-infection . Epidemics of influenza result in an increased hospital admission rate for bacterial pneumonia due to pneumococcus, Haemophilus influenzae and Staphylococcus aureus . Similarly, an increased incidence of meningococcal diseases, particularly severe forms, follows the influenza outbreaks, with a two week delay . Though the precise mechanism is not known, the depression of host's phagocytes bactericidal activity by the influenza virus seems to be involved . An increased incidence of invasive group A beta hemolytic streptococcal infections, particularly necrotizing fasciitis and toxic shock syndrome, is also observed in relation with chickenpox . The reason for this association is unclear and appears not to be limited to the disruption of the cutaneous barrier which leads to the cutaneous infections in this disease . Bacterial-viral co-infection is not a justification for a systematic antibiotic prescription in viral diseases . Severe bacterial disease will be best prevented through viral immunization, thus encouraging the development of viral vaccines and immunization campaigns.

Mol Microbiol, 1998 Feb, 27(3), 617 - 30
Secretion of the Haemophilus influenzae HMW1 and HMW2 adhesins involves a periplasmic intermediate and requires the HMWB and HMWC proteins; St Geme JW 3rd et al.; Non-typable Haemophilus influenzae is a common cause of human disease and initiates infection by colonizing the upper respiratory tract . The non-typeable H . influenzae HMW1 and HMW2 non-pilus adhesins mediate attachment to human epithelial cells, an essential step during colonization . In order to facilitate interaction with host cells, HMW1 and HMW2 are localized on the surface of the organism in a process that involves cleavage of a 441-amino-acid N-terminal fragment . In the present study, we investigated the pathway for the secretion of HMW1 and HMW2 . Cell fractionation experiments and cryoimmunoelectron microscopy demonstrated that a periplasmic intermediate occurs, suggesting involvement of the Sec machinery . Additional analysis revealed that, ultimately, the proteins are partially released from the surface of the organism . Studies with Escherichia coli harbouring plasmid subclones extended earlier findings and suggested that the secretion of HMW1 requires accessory proteins designated HMW1B and HMW1C, while the secretion of HMW2 requires proteins called HMW2B and HMW2C . Further analysis established that HMW1B/HMW1C and HMW2B/HMW2C are interchangeable, an observation consistent with the high degree of homology between HMW1B and HMW2B and between HMW1C and HMW2C . Additional studies of the hmw1 locus indicated that HMW1B is located in the outer membrane and serves to translocate HMW1 across the outer membrane . In the absence of HMW1B, HMW1 remains unprocessed and is degraded in the periplasmic space, at least in part by the DegP protease . Mutagenesis of an HMW1 N-terminal motif shared with other secreted proteins resulted in diminished processing and extracellular release, suggesting interaction of this motif with the HMW1B protein . Continued investigation of the HMW1 and HMW2 adhesins may provide general insights into protein secretion and bacterial pathogenesis.

Aust N Z J Public Health, 1997 Dec, 21(7), 735 - 8
Fragmentation of scheduled visits and missed doses among infants receiving multiple injected vaccines; Ferson MJ et al.; A retrospective cohort study was used to determine the extent to which immunisation visits due in the first year of life are split into separate visits . A one-month birth cohort of infants registered in early childhood health centres in the former Eastern Sydney Health Area was followed up when the infants were 8 to 11 months of age . A telephone questionnaire sought documented dates of each dose in the primary series of diphtheria-tetanus-pertussis (DTP), Haemophilus influenzae type b (Hib) and hepatitis B (HBV) vaccination . Of the 141 subjects, 130 had received all due doses of DTP and Hib vaccines and 63 (45 per cent) had been enrolled in the neonatal hepatitis B program . Infants in the latter group received the first DTP-Hib dose on average one week later than did those not in the hepatitis B program (DTP, P = 0.016; Hib, P = 0.047) . The greatest percentage of missed DTP or Hib doses occurred in infants not receiving HBV vaccination (7.1 per cent of doses) or those high-risk infants enrolled in the neonatal hepatitis B program (2.9 per cent) . Overall, 12 infants had 28 (6.9 per cent) of the 404 possible scheduled visits fragmented into two separate visits . In all cases, parents reported that this was at the suggestion of the general practitioner . We found no greater likelihood of fragmentation for infants who had also received hepatitis B vaccine . Only 17 infants (29 per cent) had received the third hepatitis B vaccine and DTP doses at the same visit, as recommended . These findings confirm anecdotal reports of fragmentation of scheduled visits and missed doses for infants due to receive multiple injections, and some delay in uptake among those receiving hepatitis B vaccine . Universal infant hepatitis B immunisation should not be considered until combination vaccines (which should also include a Hib component) become available in Australia.

Int J Clin Pract, 1997 Sep, 51(6), 353 - 4
Doctors' knowledge of post-splenectomy prophylaxis; Palejwala AA et al.; A questionnaire was sent to 160 hospital doctors and 200 general practitioners about post-splenectomy prophylaxis during the month of September 1995 . A total of 118 questionnaires were returned (43% hospital doctors and 25% GPs) . Most doctors (99%) knew that splenectomised patients were at risk of pneumococcal infection; 72% of hospital doctors and 50% of GPs knew of the risk of Haemophilus influenza infection but only 50% of hospital doctors and 33% of GPs knew of the same risks involving meningococcal infection and malaria . Half of the GPs were not aware of H . influenzae type B (Hib) vaccine for prophylaxis and 85% of these and 50% of hospital doctors did not know prophylaxis should be lifelong . There was no significant difference in the knowledge between hospital doctors and GPs.

Arch Microbiol, 1998 Jan, 169(1), 71 - 5
Functional expression in Escherichia coli of the Haemophilus influenzae gene coding for selenocysteine-containing selenophosphate synthetase; Wilting R et al.; The selenophosphate synthetases from several organisms contain a selenocysteine residue in their active site where the Escherichia coli enzyme contains a cysteine . The synthesis of these enzymes, therefore, depends on their own reaction product . To analyse how this self-dependence is correlated with the selenium status, e.g . after recovery from severe selenium starvation, we expressed the gene for the selenocysteine-containing selenophosphate synthetase from Haemophilus influenzae (selDHI) in an E . coli DeltaselD strain . Gene selDHI gave rise to a selenium-containing gene product and also supported - via its activity - the formation of E . coli selenoproteins . The results provide evidence either for the suppression of the UGASec codon with the insertion of an amino acid allowing the formation of a functional product or for a bypass of the selenophosphate requirement . We also show that the selenocysteine synthesis and the insertion systems of the two organisms are fully compatible despite conspicuous differences in the mRNA recognition motif.

Arch Microbiol, 1998 Jan, 169(1), 1 - 9
A 71-kDa protein from Halobacterium salinarium belongs to a ubiquitous P-loop ATPase superfamily with head-rod-tail structure; Ruepp A et al.; The nucleotide sequence of a genomic fragment from Halobacterium salinarium containing an open reading frame encoding a protein with a calculated molecular mass of 71 kDa was determined . Database searches revealed that this protein, Hp71, has similarities to eukaryotic cytoskeletal proteins . Heterologous production of Hp71 in Escherichia coli allowed the isolation of anti-Hp71 antibodies . The antibodies were used (1) to verify the production of Hp71 in H . salinarium and (2) to determine its cytoplasmic localization by immune electron microscopy . Homologous overproduction of Hp71 in H . salinarium and heterologous production in Haloferax volcanii resulted in modifications of cell morphology from rods to extended rods, and from pleiomorphic cells to rods, respectively . Structure prediction methods indicated that Hp71 has a head-rod-tail configuration, including an N-terminal domain with a nucleotide binding motif (P-loop), and an extended discontinuous coiled-coil domain of 330 amino acids . To identify related proteins, the complete genomes of Haemophilus influenzae, Mycoplasma genitalium, and Methanococcus jannaschii were searched for deduced proteins with extended coiled-coil domains . Only one or two proteins were found for each organism, showing that Hp71 is one of only a few prokaryotic intracellular proteins with extended coiled-coil domains . The phenotype upon overproduction and the similarity of Hp71 to the SMC superfamily of P-loop head-rod-tail proteins (named after SMC1, which is involved in the "stability of minichromosomes" in yeast) indicate that Hp71 might be involved in cytoskeleton formation and/or chromosome partitioning in H . salinarium.

Infect Immun, 1998 Mar, 66(3), 1252 - 7
GroEL heat shock protein of Haemophilus ducreyi: association with cell surface and capacity to bind to eukaryotic cells; Frisk A et al.; The Haemophilus ducreyi homolog of GroEL, a 58.5-kDa heat shock protein (Hsp), is a dominant protein produced not only in response to heat stress but also under in vitro growth conditions . Extracellular localization of the 58.5-kDa Hsp was investigated by whole-cell enzyme-linked immunosorbent assay (ELISA) and immunoelectron microscopy and in supernatants of washed bacteria by immunoblotting with a Haemophilus ducreyi GroEL-specific mouse monoclonal antibody (BB11) . To investigate binding of the Hsp to eukaryotic cells, the 58.5-kDa Hsp was purified by ion-exchange and size exclusion chromatography; incubated with HEp-2 cells, HeLa cells, and human fibroblasts; and then analyzed by immunoblotting . Direct involvement of the 58.5-kDa Hsp in the adherence of H . ducreyi to HEp-2 cells was investigated by using an inhibition assay . An epitope of the 58.5-kDa Hsp was detected by whole-cell ELISA on all of the strains tested, suggesting that it is associated with the cell surface . This was also supported by immunoelectron microscopy results . In supernatants of washed bacteria, the 58.5-kDa Hsp was detected by immunoblotting after 10 h of cultivation . The 58.5-kDa Hsp bound to the eukaryotic cells tested but exerted only limited (about 20%) inhibition of H . ducreyi adherence to HEp-2 cells . These results demonstrate that the 58.5-kDa Hsp of H . ducreyi is associated with the bacterial surface, binds to eukaryotic cells, and partially influences H . ducreyi adherence to HEp-2 cells, indicating possible involvement of the 58.5-kDa Hsp in the attachment of bacteria to host cells and to each other.

Infect Immun, 1998 Mar, 66(3), 899 - 906
The Haemophilus influenzae HtrA protein is a protective antigen; Loosmore SM et al.; The htrA gene from two strains of nontypeable Haemophilus influenzae has been cloned and sequenced, and the encoded approximately 46-kDa HtrA proteins were found to be highly conserved . H . influenzae HtrA has approximately 55% identity with the Escherichia coli and Salmonella typhimurium HtrA stress response proteins, and expression of the H . influenzae htrA gene was inducible by high temperature . Recombinant HtrA (rHtrA) was expressed from E . coli, and the purified protein was found to have serine protease activity . rHtrA was found to be very immunogenic and partially protective in both the passive infant rat model of bacteremia and the active chinchilla model of otitis media . Immunoblot analysis indicated that HtrA is antigenically conserved in encapsulated and nontypeable H . influenzae species . Site-directed mutagenesis was performed on the htrA gene to ablate the endogenous serine protease activity of wild-type HtrA, and it was found that eight of nine recombinant mutant proteins had no measurable residual proteolytic activity . Two mutant proteins were tested in the animal protection models, and one, H91A, was found to be partially protective in both models . H91A HtrA may be a good candidate antigen for a vaccine against invasive H . influenzae type b disease and otitis media and is currently in phase I clinical trials.

Clin Exp Immunol, 1998 Feb, 111(2), 237 - 42
Subclass distribution of IgA antibodies in saliva and serum after immunization with Haemophilus influenzae type b conjugate vaccines; Kauppi-Korkeila M et al.; IgA subclass distribution of antibodies against capsular polysaccharide (PS) of Haemophilus influenzae type b (Hib) was studied in saliva and serum samples of children vaccinated with two (n = 58) or three doses (n = 53) of Hib vaccine . One month after the second dose of Hib conjugate vaccine, at 7 months old, 40% of the children had IgA1 and 41% had IgA2 anti-Hib PS antibodies in saliva . One month after the third dose, at 15-25 months old, IgA1 was the predominating subclass; 72% of the children had IgA1, 26% had IgA2 anti-Hib PS in saliva . The mean concentration of IgA1 anti-Hib PS, expressed as optical density (OD) values, was significantly higher after three doses (OD 80.7) than after two doses (OD 18.9) . The mean concentration of IgA2 did not change significantly after the third dose (OD 23.8 after two doses, OD 18.1 after three doses) . In serum, IgA1 anti-Hib PS predominated both after two (17% had IgA1, none had IgA2) and three doses (72% had IgA1, 4% had IgA2) of Hib vaccine . In conclusion, both IgA1 and IgA2 anti-Hib PS were found in saliva of immunized children after two doses of Hib conjugate vaccine, whereas the third vaccine dose induced a shift towards IgA1 anti-Hib PS dominance in saliva.

Mol Microbiol, 1998 Jan, 27(2), 391 - 404
Periplasmic copper-zinc superoxide dismutase protects Haemophilus ducreyi from exogenous superoxide; San Mateo LR et al.; Haemophilus ducreyi causes chancroid, a sexually transmitted genital ulcer disease implicated in increased heterosexual transmission of HIV . As part of an effort to identify H . ducreyi gene products involved in virulence and pathogenesis, we created random TnphoA insertion mutations in an H . ducreyi 35000 library cloned in Escherichia coli . Inserts encoding exported or secreted PhoA fusion proteins were characterized by DNA sequencing . One such clone encoded a Cu-Zn superoxide dismutase (SOD) enzyme . The Cu-Zn SOD was periplasmic in H . ducreyi and accounted for most of the detectable SOD activity in whole-cell lysates of H . ducreyi grown in vitro . To investigate the function of the Cu-Zn SOD, we created a Cu-Zn SOD-deficient H . ducreyi strain by inserting a cat cassette into the sodC gene . The wild-type and Cu-Zn SOD null mutant strains were equally resistant to excess cytoplasmic superoxide induced by paraquat, demonstrating that the Cu-Zn SOD did not function in the detoxification of cytoplasmic superoxide . However, the Cu-Zn SOD null strain was significantly more susceptible to killing by extracellular superoxide than the wild type . This result suggests that the H . ducreyi Cu-Zn SOD may play a role in bacterial defence against oxidative killing by host immune cells during infection.

J Antimicrob Chemother, 1997 Dec, 40 Suppl A, 73 - 81
Randomized, double-blind, comparative study of grepafloxacin and amoxycillin in the treatment of patients with community-acquired pneumonia; O'Doherty B et al.; This randomized, multicentre, double-blind, double-dummy study assessed the efficacy and safety of 7 or 10 day regimens of grepafloxacin, 600 mg od, compared with amoxycillin, 500 mg tds, in the treatment of community-acquired pneumonia (CAP) . A total of 264 patients were recruited at 43 centres (127 received grepafloxacin and 137 received amoxycillin), of whom 207 patients (78%) completed the study . Clinical and microbiological efficacy were assessed at the end-of-treatment visit (3-5 days after the last dose) and at the follow-up visit (28-42 days after the last dose) . At follow-up, patients in the evaluable population treated with grepafloxacin demonstrated a clinical response rate (76%; 87/114) equivalent to that seen with amoxycillin (74%, 85/111, 95% CI = -12%, 10%) while, in the intent-to-treat population with a documented bacterial pathogen, the clinical success rate in the grepafloxacin group (78%, 29/37) was significantly higher than in the amoxycillin group (58%, 28/48), 95% CI = 2%, 43%) . In patients from the evaluable population in whom the pathogens were documented the clinical success rate favoured grepafloxacin, compared with amoxycillin (79%, 26/33 versus 63%, 26/42, respectively; 95% CI = -5.2%, 38.1%) . Microbiological eradication with grepafloxacin was statistically superior to amoxycillin in the evaluable population; the success rate was 89% (32/36) in the grepafloxacin group compared with 71% (32/45) for the amoxycillin group (95% CI = 2%, 37%) . The pathogens most commonly isolated from patients were Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae . The success rates for infections caused by S . pneumoniae and H . influenzae at follow-up were higher with grepafloxacin than with amoxycillin . Grepafloxacin was well tolerated, with a safety profile comparable to that of amoxycillin . The therapeutic judgement of patients and investigators at the patient's last visit, as well as the assessment of individual respiratory signs and symptoms, yielded comparable results with both treatments . The results of this study indicate that grepafloxacin, 600 mg od for 7-10 days, is equivalent to or better than amoxycillin, 500 mg tds for 7-10 days in achieving a successful clinical and microbiological response in the treatment of patients with CAP.

J Antimicrob Chemother, 1997 Dec, 40 Suppl A, 63 - 72
Randomized, double-blind study of grepafloxacin versus amoxycillin in patients with acute bacterial exacerbations of chronic bronchitis; Langan CE et al.; This randomized, multicentre, double-blind, double-dummy study compared the efficacy and safety of grepafloxacin and amoxycillin in acute bacterial exacerbations of chronic bronchitis (ABECB) . Patients were randomized to receive grepafloxacin 400 mg or 600 mg od, or amoxycillin 500 mg tds, for 7 or 10 days . The trial recruited 656 patients, of whom 566 (86%) completed the study . Clinical success rates at the 2 week follow-up visit for the population evaluable for clinical efficacy were 82% (165/202 patients) in the grepafloxacin 400 mg group, 85% (175/206) in the grepafloxacin 600 mg group and 85% (172/203 patients) in the amoxycillin group . The 95% confidence interval confirmed the equivalence of the two grepafloxacin doses and amoxycillin, with no significant difference between the grepafloxacin groups . The microbiological success rates at follow-up showed equivalence between the grepafloxacin 400 mg and amoxycillin groups: 86% (144/168 isolates) and 83% (162/195), respectively . The grepafloxacin 600 mg group achieved a statistically significantly higher eradication rate (92%, 150/164; 95% CI 2.0%, 16.1%) than the amoxycillin group in the follow-up assessment for microbiological and clinical efficacy (evaluable population) . There was no significant difference between the two grepafloxacin treatment groups (95% CI -13.3%, 0.9%; P= 0.087) . All three treatment regimens successfully eradicated the pathogens most commonly isolated during the study, including Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae . Grepafloxacin had a good safety profile, comparable to that of amoxycillin, although grepafloxacin 600 mg was associated with a higher incidence of nausea, dyspepsia and taste perversion than amoxycillin . It can be concluded that grepafloxacin 400 mg or 600 mg od is as effective as amoxycillin 500 mg tds in the treatment of ABECB.

J Antimicrob Chemother, 1997 Dec, 40 Suppl A, 27 - 30
The activity of grepafloxacin against respiratory pathogens in the UK; Wise R et al.; The in-vitro activity of grepafloxacin was compared with that of other antimicrobials against respiratory tract pathogens collected from 15 UK laboratories over the winter of 1995-96 . Penicillin-resistant Streptococcus pneumoniae was not encountered, but macrolide resistance was seen in approximately 10% of strains . Grepafloxacin (MIC90 0.25 mg/L) was four- to eight-fold more active than ciprofloxacin . Twelve percent of Haemophilus influenzae were beta-lactamase producers, macrolides were relatively inactive yet fluoroquinolones were highly active . Moraxella catarrhalis were highly susceptible to fluoroquinolones and macrolides . The activity of grepafloxacin against respiratory tract pathogens should make it a useful agent in the treatment of infections at this site.

Semin Oncol, 1998 Feb, 25(1), 98 - 106
The infectious complications of chronic lymphocytic leukemia; Morrison VA; Infectious complications continue to have a major impact on the clinical course of patients with chronic lymphocytic leukemia despite advances in therapeutic approaches to this disease and supportive care . Although the pathogenesis of infection in these patients is multifactorial, systemic hypogammaglobulinemia is the major immune defect accounting for the increased risk of infection . Despite common knowledge of systemic immune defects in this population, information regarding mucosal immune function is minimal . In patients treated with conventional alkylating agents, infections commonly occur at mucosal sites, especially the respiratory tract, and organisms such as Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa are frequent isolates . The use of purine analogues as fludarabine has resulted in a change in this spectrum of infection, with the appearance of opportunistic infections caused by Pneumocystis, Listeria, Mycobacterium tuberculosis, Nocardia, Candida, Aspergillus, and herpesviruses . Further knowledge of the impact of chemotherapy on immune function, and of the immune defects in these patients, both inherent to the primary disease process and therapy-related, will aid in the formulation of better prophylactic and therapeutic interventions to reduce the risk of infection and improve the ultimate outcome of patients with chronic lymphocytic leukemia.

Nucleic Acids Res, 1998 Jan 15, 26(2), 544 - 8
Microbial gene identification using interpolated Markov models; Salzberg SL et al.; This paper describes a new system, GLIMMER, for finding genes in microbial genomes . In a series of tests on Haemophilus influenzae , Helicobacter pylori and other complete microbial genomes, this system has proven to be very accurate at locating virtually all the genes in these sequences, outperforming previous methods . A conservative estimate based on experiments on H.pylori and H . influenzae is that the system finds >97% of all genes . GLIMMER uses interpolated Markov models (IMMs) as a framework for capturing dependencies between nearby nucleotides in a DNA sequence . An IMM-based method makes predictions based on a variable context; i.e., a variable-length oligomer in a DNA sequence . The context used by GLIMMER changes depending on the local composition of the sequence . As a result, GLIMMER is more flexible and more powerful than fixed-order Markov methods, which have previously been the primary content-based technique for finding genes in microbial DNA.

Ann Pharmacother, 1998 Jan, 32(1), S27 - 30
Novel cost-effective approaches to the treatment of community-acquired infections; Schwarzmann SW; OBJECTIVE: To review approaches to the diagnosis and treatment of patients with community-acquired infections . INTRODUCTION: Dramatic changes in the antibiotic susceptibility of pathogens commonly associated with community-acquired infections have occurred during the past decade . DISCUSSION: Changes in the antibiotic sensitivity profile of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis that have occurred over the past several years have required modifications in empiric antibiotic selections for infections due to these pathogens . The most profound changes have occurred with S . pneumoniae, which has shown significant resistance to beta-lactams by means of alteration of one or more of the five important penicillin-binding proteins . Many of these organisms have become resistant to other classes of antibiotics; some are sensitive only to vancomycin . H . influenzae and M . catarrhalis have developed resistance primarily by production of beta-lactamase . CONCLUSIONS: The antibiotic selection process for the treatment of community-acquired pneumonia relates to the site of infection and, in many cases, the in vitro sensitivity testing results or known patterns in a given geographic area.

Viral Immunol, 1997, 10(4), 197 - 206
Correlation of in vitro T-cell and B-cell function with responses to childhood vaccines in children with human immunodeficiency virus infection; Peters VB et al.; We analyzed T-cell responses to mitogens and antigens and B-cell differentiation in response to T-cell dependent (TCD) and independent stimuli in 22 human immunodeficiency virus (HIV)-infected children (1 to 9 years of age) according to the presence of protective humoral immunity at a mean time of 18 months after vaccination with Haemophilus influenzae type b, hepatitis B, diphtheria, and tetanus vaccines . The 17 vaccine responders had a mean of 3.2 responses . However, their antibody levels were lower compared with healthy children . The 5 nonresponders had a mean of 0.84 responses . There were no significant differences between responders and nonresponders regarding age, Centers for Disease Control and Prevention (CDC) disease class, CDC immunologic class, serum immunoglobulin (Ig) levels, or in the use of antiretroviral therapy . However, responders tended to have higher age-adjusted absolute CD4 cell counts than nonresponders (p = 0.07) . Nonetheless, there was no correlation between antibody levels and age-adjusted CD4 counts for each of the 4 TCD vaccines . Responders had conserved lymphoproliferative responses to mitogens and to candida antigen; 7 (41%) had normal responses to tetanus antigen . While nonresponders had some conserved responses to mitogens, only 1 had a response to antigen . Thirteen responders (77%) and only 1 nonresponder (20%) had normal responses to at least 2 of the 3 mitogens and 1 of the 2 antigens (p = 0.04) . Although defects in B-cell differentiation were detected in both groups, they were profound and generalized in the nonresponders . Fourteen responders (82%) had at least 1 normal B-cell response compared with none of the 5 nonresponders (p = 0.002) . There were no correlations between normal lymphoproliferative responses and age, CD4 counts, serum immunoglobulin G (IgG) levels, or the use of antiretroviral therapy . Immunologic function is important in the evaluation of HIV-infected children.

Acta Otorrinolaringol Esp, 1997 Aug-Sep, 48(6), 479 - 82
{The clinical importance of bacteremia during tonsillectomy}; Soldado L et al.; INTRODUCTION: Post-tonsillectomy bacteraemia is a well recognized aetiological factor in streptococcal endocarditis . Prophylactic penicillin has been recommended to reduce its incidence in susceptible patients undergoing tonsillectomy . Recent studies have shown a change in the microflora and an increase in the number of penicillin-resistant organisms in the tonsils of patients undergoing tonsillectomy . OBJECTIVE: The aim of this study was to assess the incidence of post-tonsillectomy bacteraemia, identify the associated organisms, and review the suitability of penicillin in prophylactic regimens . MATERIAL AND METHODS: 102 children were included . Blood culture samples were taken after removal of the first tonsil, which was randomly electrodissected or blunt dissected . Statistical analysis was performed using ANOVA . RESULTS: Of the 102 patients included in the study, 41 (40.1%) had positive post-tonsillectomy blood cultures . Haemophilus influenzae was isolated from 23 (56%) of the positive cultures and Streptococcus viridans from 15 (36.5%) . Twenty-five percent of H . influenzae and 50% of the viridans group produced beta-lactamase . CONCLUSIONS: A beta-lactamase stable antibiotic would be more appropriate than penicillin for prophylaxis during tonsillectomy.

Pediatr Infect Dis J, 1998 Jan, 17(1), 23 - 8
Epidemiology of invasive pneumococcal disease in the Western Region, The Gambia; Usen S et al.; BACKGROUND: Streptococcus pneumoniae is a major cause of morbidity and mortality in young children in the developing world . The recent development of pneumococcal polysaccharide/protein conjugate vaccines may make possible prevention of this infection . However, little is known about the epidemiology of invasive pneumococcal disease in children in the developing world . OBJECTIVES: To determine the incidence and epidemiologic features of invasive pneumococcal disease in children resident in a semiurban area of The Gambia . METHOD: The study was part of a large trial of an Haemophilus influenzae type b vaccine that recruited 42 848 children at the age of 2 months during the period March, 1993, to October, 1995 . Follow-up of study children continued until December 31, 1995; therefore the first children to enter the trial were followed for 2.5 years and the last for just a few months . During the period of surveillance, 2256 children were investigated for possible invasive pneumococcal disease when they presented to a hospital or health center . RESULTS: We detected 110 cases of pneumococcal disease . Pneumonia was the most common form of invasive pneumococcal disease observed (75.5% of patients) . The incidence of pneumococcal disease was 224 {95% confidence interval (CI) 171, 277} per 100,000 child years among children ages 2 to 11 months, 139 (95% CI 93, 184) per 100,000 among children ages 12 to 23 months and 82 (95% CI 21, 143) per 100,000 among children ages 24 to 35 months . Pneumococci of serogroups 14, 6, 5, 23, 19, 46 and 2 were isolated most frequently . Susceptibility to pneumococcal disease was not increased significantly among Haemophilus influenzae type b-vaccinated children . CONCLUSIONS: The pneumococcus is a major cause of bacterial infection in The Gambia . A proposed nine-valent pneumococcal conjugate vaccine for developing countries containing conjugates of serogroups 1, 4, 5, 6, 9, 14, 18, 19 and 23 would cover 74% of cases of invasive pneumococcal disease in children resident in the Western Region of The Gambia.

Am J Ophthalmol, 1998 Feb, 125(2), 261 - 3
Extrusion of an anterior chamber intraocular lens complicated by Haemophilus endophthalmitis; Lewis JM et al.; PURPOSE: To describe a case of late postoperative endophthalmitis in a patient who had undergone anterior chamber intraocular lens implantation 6 years previously . METHOD: Onset, course, and outcome of the patient's eye disease are presented . RESULTS: The patient was initially examined with erosion of the eyewall, extrusion of the lens haptic, and endophthalmitis . Intraocular cultures disclosed the pathogenic organism to be Haemophilus influenzae . CONCLUSION: Anterior chamber intraocular lens haptic extrusion can have grave consequences, including endophthalmitis and severe visual loss.

Oral Microbiol Immunol, 1997 Jun, 12(3), 174 - 7
Cloning and sequencing of part of the S10 operon from Actinobacillus actinomycetemcomitans FDC Y4; Hayashida H et al.; We have cloned and sequenced the 5.2 kb EcoRI fragment that contained part of the S10 operon from Actinobacillus actinomycetemcomitans FDC Y4 . The order of the ribosomal protein genes was identical to that of the S10 operon of Haemophilus influenzae and Escherichia coli . The deduced amino acid sequences of ribosomal proteins in this operon displayed significant homologies (65.3%-100%) to those of H . influenzae, E . coli, Yersinia enterocolitica and Yersinia pseudotuberculosis . Phylogenetic trees obtained for these ribosomal proteins were similar to that obtained for 16S rRNA.

J Clin Microbiol, 1998 Feb, 36(2), 539 - 42
Viruses and bacteria in the etiology of the common cold; Makela MJ et al.; Two hundred young adults with common colds were studied during a 10-month period . Virus culture, antigen detection, PCR, and serology with paired samples were used to identify the infection . Viral etiology was established for 138 of the 200 patients (69%) . Rhinoviruses were detected in 105 patients, coronavirus OC43 or 229E infection was detected in 17, influenza A or B virus was detected in 12, and single infections with parainfluenza virus, respiratory syncytial virus, adenovirus, and enterovirus were found in 14 patients . Evidence for bacterial infection was found in seven patients . Four patients had a rise in antibodies against Chlamydia pneumoniae, one had a rise in antibodies against Haemophilus influenzae, one had a rise in antibodies against Streptococcus pneumoniae, and one had immunoglobulin M antibodies against Mycoplasma pneumoniae . The results show that although approximately 50% of episodes of the common cold were caused by rhinoviruses, the etiology can vary depending on the epidemiological situation with regard to circulating viruses . Bacterial infections were rare, supporting the concept that the common cold is almost exclusively a viral disease.

J Infect, 1997 Nov, 35(3), 304 - 8
Haemophilus aphrophilus bacteraemia complicated with vertebral osteomyelitis and spinal epidural abscess in a patient with liver cirrhosis; Hung CC et al.; Haemophilus aphrophilus is rarely implicated as an aetiology of spinal epidural abscess . A 73-year-old woman with liver cirrhosis who developed H . aphrophilus bacteraemia complicated with vertebral osteomyelitis and spinal epidural abscess is presented . Without surgical decompression, she was successfully treated with cefotaxime for 3 weeks, followed by maintenance with ciprofloxacin for another 10 weeks . The clinical features of eight previously reported cases of vertebral osteomyelitis without epidural abscess due to H . aphrophilus are reviewed.

Diagn Microbiol Infect Dis, 1997 Dec, 29(4), 249 - 57
Surveillance of antimicrobial resistance in Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis in the United States in 1996-1997 respiratory season . The Laboratory Investigator Group; Thornsberry C et al.; A U.S . surveillance study of antimicrobial resistance in respiratory tract pathogens in the respiratory season (1996-1997) is reported that included 11,368 isolates from 434 institutions in 45 states and the District of Columbia . beta-lactamase was produced by 33.4% of Haemophilus influenzae and 92.7% of Moraxella catarrhalis . Of the 9,190 Streptococcus pneumoniae isolates tested, 33.5% were not susceptible to penicillin (MIC > or = 0.12 microgram/mL), with 13.6% having high-level resistance (MICs > 1 microgram/mL) . For H . influenzae, the most active antimicrobials (based on percent of strains susceptible) were levofloxacin (100%) and ceftriaxone (99.9%); the least active were ampicillin (67.2%) and clarithromycin (58.1%) . For M . catarrhalis, the most active drugs were amoxicillin-clavulanate, ceftriaxone, and levofloxacin (100%); the least active was ampicillin . The order of the activity of the drugs against S . pneumoniae were levofloxacin (97.3%) > ceftriaxone (87.1%) > amoxicillin-clavulanate (81.7%) = clarithromycin (80.9%) > cefuroxime (74.5%) > penicillin (66.5%) . The activity of the beta-lactams and clarithromycin against isolates of S . pneumoniae was closely associated with the resistance to penicillin . Levofloxacin was more active against S . pneumoniae overall, because it exhibited no cross-resistance . These data indicate that the incidence of beta-lactamase production in H . influenzae (33.4%) and M . catarrhalis (92.7%) is similar to other recent studies, and that the incidence of penicillin-intermediate and -resistant S . pneumoniae is increasing, particularly the high-level penicillin-resistant (MICs > 1 microgram/mL) strains, which were often multi-resistant.

N Engl J Med, 1997 Oct 2, 337(14), 970 - 6
Bacterial meningitis in the United States in 1995 . Active Surveillance Team; Schuchat A et al.; BACKGROUND: Before the introduction of the conjugate vaccines, Haemophilus influenzae type b was the major cause of bacterial meningitis in the United States, and meningitis was primarily a disease of infants and young children . We describe the epidemiologic features of bacterial meningitis five years after the H . influenzae type b conjugate vaccines were licensed for routine immunization of infants . METHODS: Data were collected from active, population-based surveillance for culture-confirmed meningitis and other invasive bacterial disease during 1995 in laboratories serving all the acute care hospitals in 22 counties of four states (total population, more than 10 million) . The rates were compared with those for 1986 obtained by similar surveillance . RESULTS: On the basis of 248 cases of bacterial meningitis in the surveillance areas, the rates of meningitis (per 100,000) for the major pathogens in 1995 were Streptococcus pneumoniae, 1.1; Neisseria meningitidis, 0.6; group B streptococcus, 0.3; Listeria monocytogenes, 0.2; and H . influenzae, 0.2 . Group B streptococcus was the predominant pathogen among newborns, N . meningitidis among children 2 to 18 years old, and S . pneumoniae among adults . Pneumococcal meningitis had the highest case fatality rate (21 percent) and in 36 percent of cases was caused by organisms that were not susceptible to penicillin . From these data, we estimate that 5755 cases of bacterial meningitis were caused by these five pathogens in the United States in 1995, as compared with 12,920 cases in 1986, a reduction of 55 percent . The median age of persons with bacterial meningitis increased greatly, from 15 months in 1986 to 25 years in 1995, largely as a result of a 94 percent reduction in the number of cases of H . influenzae meningitis . CONCLUSIONS: Because of the vaccine-related decline in meningitis due to H . influenzae type b, bacterial meningitis in the United States is now a disease predominantly of adults rather than of infants and young children.

J Photochem Photobiol B, 1997 Oct, 40(3), 204 - 8
Photodynamic destruction of Haemophilus parainfluenzae by endogenously produced porphyrins; van der Meulen FW et al.; Bacterial resistance against antibiotic treatment is becoming an increasing problem in medicine . Therefore methods to destroy microorganisms by other means are being investigated, one of which is photodynamic therapy (PDT) . It has already been shown that a variety of Gram-positive and Gram-negative bacteria can be killed in vitro by PDT using exogenous sensitizers . An alternative method of photosensitizing cells is to stimulate the production of endogenous sensitizers . The purpose of this study was to investigate the bactericidal efficacy of PDT for Haemophilus parainfluenzae with endogenously produced porphyrins, synthesized in the presence of delta-aminolaevulinic acid (delta-ALA) . H . parainfluenzae incubated with increasing amounts of delta-ALA showed decreased survival after illumination with 630 nm light . No photodynamic effect on the bacterial viability was found when H . parainfluenzae was grown without added delta-ALA . H . influenzae, grown in the presence of delta-ALA, but not capable of synthesizing porphyrins from delta-ALA, was not affected by PDT . Of the range of incident wavelengths, 617 nm appeared to be the most efficient in killing the bacteria . Spectrophotometry of the bacterial porphyrins demonstrated that the maximum fluorescence occurred at approximately 617 nm, with a much lower peak around 680 nm . We conclude that a substantial killing of H . parainfluenzae by PDT in vitro after endogenous sensitization with delta-ALA can be achieved.

J Bacteriol, 1998 Feb, 180(3), 746 - 8
A new transformation-deficient mutant of Haemophilus influenzae Rd with normal DNA uptake; Gwinn ML et al.; Haemophilus influenzae Rd is a gram-negative natural transformer . A mutant strain, RJ248, that has normal DNA uptake and translocation but whose transformation frequency is 300 times lower than that of wild-type H . influenzae and whose phage recombination is 8 times lower was isolated . The affected gene, comM, is induced during competence development in wild-type H . influenzae but not in RJ248.

Rev Med Chir Soc Med Nat Iasi, 1996 Jul-Dec, 100(3-4), 128 - 34
{The postantibiotic effect of azithromycin on respiratory pathogens}; Diculencu D et al.; Azithromycin has in vitro activity which includes important respiratory pathogens and is successful in treatment of respiratory tract infections . We assessed postantibiotic effect (PAE) of azithromycin against 3 stains of Streptococcus pneumoniae, 2 strains of Haemophilus influenzae and 2 strains of Moraxella catarrhalis . The strains were exposed for 2 hours to an azithromycin concentration of 0.5 mg/L (maximum serum concentration achieved by azithromycin after the usual dosing regimen) . A stationary phase inoculum of 1 x 10(6)-5 x 10(6) UFC/ml in IsoSensitest Broth with 5% lysed horse blood and 20 mg/L NAD was used and shaken for the duration of the experiment . Antibiotic was neutralised by dilution 1:1000 into pre-warmed medium . Viable counts were determined before and after antibiotic exposure and then hourly for 7 hours by Miles and Misra method . The experiment was performed in triplicate . Even at such low concentration as that achieved in serum, azithromycin exhibits a PAE of 119 min for pneumococci, 130 min for haemophili and 155 min for moraxellae, fact which could allow the use of usual oral regimen in bacteraemic respiratory infection, as well.

Infect Immun, 1998 Feb, 66(2), 656 - 63
Comparative analysis of Haemophilus influenzae hifA (pilin) genes; Clemans DL et al.; Adherence of Haemophilus influenzae to epithelial cells plays a central role in colonization and is the first step in infection with this organism . Pili, which are large polymorphic surface proteins, have been shown to mediate the binding of H . influenzae to cells of the human respiratory tract . Earlier experiments have demonstrated that the major epitopes of H . influenzae pili are highly conformational and immunologically heterogenous; their subunit pilins are, however, immunologically homogenous . To define the extent of structural variation in pilins, which polymerize to form pili, the pilin genes (hifA) of 26 type a to f and 16 nontypeable strains of H . influenzae were amplified by PCR and subjected to restriction fragment length polymorphism (RFLP) analysis with AluI and RsaI . Six different RFLP patterns were identified . Four further RFLP patterns were identified from published hifA sequences from five nontypeable H . influenzae strains . Two patterns contained only nontypeable isolates; one of these contained H . influenzae biotype aegyptius strains F3031 and F3037 . Another pattern contained predominantly H . influenzae type f strains . All other patterns were displayed by a variety of capsular and noncapsular types . Sequence analysis of selected hifA genes confirmed the 10 RFLP patterns and showed strong identity among representatives displaying the same RFLP patterns . In addition, the immunologic reactivity of pili with antipilus antisera correlated with the groupings of strains based on hifA RFLP patterns . Those strains that show greater reactivity with antiserum directed against H . influenzae type b strain M43 pili tend to fall into one RFLP pattern (pattern 3); while those strains that show equal or greater reactivity with antiserum directed against H . influenzae type b strain Eagan pili tend to fall in a different RFLP pattern (pattern 1) . Sequence analysis of representative HifA pilins from typeable and nontypeable H . influenzae identified several highly conserved regions that play a role in bacterial pilus assembly and other regions with considerable amino acid heterogeneity . These regions of HifA amino acid sequence heterogeneity may explain the immunologic diversity seen in intact pili.

Infect Immun, 1998 Feb, 66(2), 406 - 17
The fimbria gene cluster of nonencapsulated Haemophilus influenzae; Geluk F et al.; The occurrence of fimbria gene clusters in nonencapsulated Haemophilus influenzae strains from chronic bronchitis patients (n = 58), patients with acute otitis media (n = 13), and healthy carriers (n = 12) was determined by DNA hybridization and PCR, based on sequences of fimbriate H . influenzae type b . Although an average of 18% of all nonencapsulated strains had a fimbria gene cluster consisting of hifA to hifE inserted in the chromosome between purE and pepN, differences in the frequency of fimbria cluster-positive strains were observed, depending on the source of isolates . The compositions of the fimbria gene clusters of seven strains from chronic bronchitis patients and one strain from an otitis media patient were analyzed in more detail . After enrichment for fimbria expression, the promoter of the gene cluster contained 10 TA repeats (n = 2), leading to optimal positioning between the -10 and -35 promoter regions . The promoter regions of five fimbria-negative strains were sequenced; four were found to have nine TA repeats, and one had only four TA repeats . The protein sequence of three ganglioside GM1-specific HifA adhesins consisted of conserved regions intermingled with regions of sequence diversity . hifA appeared to be flanked by intergenic regions that varied between strains and contained both direct and inverted DNA repeats . Since noncoding DNA between hifA and purE has not been found in H . influenzae type b, these DNA sequences are probably not essential for fimbria expression . An analysis of strains lacking the gene cluster revealed the presence of similar sequences in 13 of 15 strains from chronic bronchitis patients, 5 of 5 strains from otitis media patients, and 3 of 5 strains from healthy carriers . The lengths of these intergenic regions were the same for multiple isolates of strains obtained during persistent infections . The presence or absence and the composition of the fimbria gene cluster and other sequences between the flanking genes purE and pepN suggest that the fimbria gene cluster was originally contained on a mobile element.

FEMS Immunol Med Microbiol, 1997 Nov, 19(3), 231 - 6
Characterisation of an outer membrane protein of Moraxella catarrhalis; Mathers KE et al.; To elucidate potential vaccine antigens, Moraxella catarrhalis outer membrane proteins (OMPs) were studied . We have previously shown an OMP to be a target for human IgG and have now further characterised this OMP which appears to have a molecular mass of 84 kDa and to be distinct from the 81-kDa OMP, CopB . Human transferrin was shown to bind the 84-kDa OMP alone . N-terminal sequencing of this OMP and purified M . catarrhalis transferrin binding protein B (TbpB) revealed homology both with each other and with the TbpB of Haemophilus influenzae and Neisseria meningitidis . Adsorption of human anti-serum with purified TbpB from two M . catarrhalis strains abolished or reduced binding of IgG to the 84-kDa OMP from three M . catarrhalis isolates . IgG binding to CopB was unaffected . It is clear that the 84-kDa OMP is distinct from CopB and is a likely homologue of TbpB.

FEMS Microbiol Lett, 1998 Jan 1, 158(1), 57 - 60
Construction of antibiotic resistance cassettes with multiple paired restriction sites for insertional mutagenesis of Haemophilus influenzae; Whitby PW et al.; Insertional mutagenesis of cloned genes coupled with site specific recombination into the genome of the parent organism is an ideal method for characterizing gene function . In this paper we describe the production and utility of two antibiotic resistance cassettes for use in Haemophilus influenzae . The mutagenic elements encode resistance to chloramphenicol or spectinomycin . Multiple paired restriction enzyme sites bound both cassettes . Use of these constructs to create mutants in H . influenzae demonstrated that the cassettes are readily incorporated into the genome in single copy and allow easy detection of mutant constructs . The insertions are stable following repeated in vitro passage . In addition, the elements are compatible with each other and allow the construction of multiple mutations within a single strain.

Vet Microbiol, 1997 Nov, 58(2-4), 261 - 76
Characterization of H . parasuis periplasmic nucleotide pyrophosphatase as a potential target enzyme for inhibition of growth; Wise DJ et al.; The periplasmic nucleotide pyrophosphatase from Haemophilus parasuis was purified 750-fold to electrophoretic homogeneity through salt fractionation and ion-exchange and affinity chromatography . The purified enzyme was monomeric with an apparent M(r) of 70,000 and catalyzed the hydrolysis of the pyrophosphate bond of NAD to yield NMN and AMP as products . The enzyme exhibited negative cooperativity in the hydrolysis of a number of pyridine dinucleotides and structurally-related pyrophosphate compounds as indicated by biphasic double-reciprocal plots and Hill coefficients of 0.5 . The kinetic parameters, K(m) and Vm, determined titrimetrically and analyzed through computer programs, were used to compare the relative effectiveness of dinucleotides containing nitrogen bases other than nicotinamide or adenine to that of NAD . Effective substrate-competitive inhibition of the pyrophosphatase was observed with purine and pyrimidine nucleoside diphosphates in the low micromolar concentration range . Although less effective, N1-alkylnicotinamide chlorides also inhibited competitively with respect to the substrate, NAD . In addition to being an effective inhibitor of the purified enzyme, adenosine diphosphate also inhibited growth of H . parasuis at a low micromolar concentration . This inhibition of growth correlates well with inhibition of the periplasmic pyrophosphatase which is supported by the fact that adenosine diphosphate does not effectively inhibit growth when the pyrophosphatase is by-passed by growth on nicotinamide mononucleotide . These observations are all consistent with the periplasmic nucleotide pyrophosphatase being essential for the growth of the organism on NAD and therefore, a very important enzyme with respect to the pathogenesis of the organism . 3-Aminopyridine mononucleotide, which also inhibited growth of H . parasuis at a low micromolar concentration, did not effectively inhibit the purified pyrophosphatase and a different target enzyme needs to be considered to explain growth inhibition by this derivative.

Spine, 1997 Dec 1, 22(23), 2763 - 5
Spinal abscess of Haemophilus paraphrophilus . A case report; Samuel W et al.; STUDY DESIGN: A case of paraspinal abscess formation from Haemophilus paraphrophilus is presented . OBJECTIVES: To describe a case of paraspinal abscess formation from H . paraphrophilus, a fastidious commensal organism of the mouth and pharynx . A precise bacteriologic identification can be difficult; techniques for such identification are discussed . SUMMARY OF BACKGROUND DATA: Spinal abscess caused by H . paraphrophilus is unusual and can be very difficult to diagnose . METHODS: The etiology, clinical presentation, technical examinations, and treatment are reviewed . RESULTS: Prolonged antibiotic treatment was curative, although surgery was considered . CONCLUSIONS: Bacteriologic diagnoses in these rare infections are difficult . Antibiotic therapy was curative in the patient described.

Clin Infect Dis, 1997 Dec, 25(6), 1378 - 84
Administration of combined diphtheria and tetanus toxoids and pertussis vaccine, hepatitis B vaccine, and Haemophilus influenzae type b (Hib) vaccine to infants and response to a booster dose of Hib conjugate vaccine; Pichichero ME et al.; We compared antibody levels following separate but simultaneous administration of diphtheria and tetanus toxoids with acellular pertussis vaccine (DTaP) containing pertussis toxoid, filamentous hemagglutinin, and pertactin (PRN); hepatitis B vaccine; and Haemophilus influenzae type b polysaccharide (polyribosylribitol phosphate; PRP) vaccine conjugated to tetanus toxoid (PRP-T) with those following administration of a combination of a DTaP-hepatitis B vaccine-PRP-T to infants at 2, 4, and 6 months of age . The antibody response to a booster dose of PRP conjugate vaccine (CRM197-OS) in infants with low (< 1 microgram/mL) or undetectable (< 0.10 microgram/mL) postpriming levels of antibody to PRP was also studied . Antibody levels were quantitated before and after dose 3 by enzyme-linked immunosorbent assay, radioimmunoassay, or neutralization assay . Seroresponse rates were not different between the two vaccine groups except for rates of response to PRP . There was a trend that levels of antibody to all the antigens included in the combination vaccine were lower than those of antibody to antigens in separate vaccines; for levels of antibody to diphtheria toxoid (P = .001), PRN (P < .0001), and PRP (P < .0001), the differences were significant . Despite low or undetectable postpriming levels of antibody to PRP, high-titered (geometric mean concentration, 9.02 micrograms/mL; range, 1.0-81.5 micrograms/mL), immunoglobulin G-predominant antibody to PRP was produced following a booster dose of CRM197-OS, a finding consistent with a memory response.

JAMA, 1998 Jan 28, 279(4), 296 - 9
Evidence of bacterial metabolic activity in culture-negative otitis media with effusion; Rayner MG et al.; CONTEXT: Otitis media with effusion (OME) can lead to significant hearing loss in children . Although previous studies have shown that bacterial DNA is present in a significant percentage of effusions sterile by culture, whether the DNA represents viable organisms or "fossilized remains" is unknown . OBJECTIVE: To determine if bacterial messenger RNA (mRNA), as detected by a reverse transcriptase-polymerase chain reaction (RT-PCR)-based assay, is present in chronic pediatric middle ear effusions that contain bacterial DNA but are sterile by standard cultural methods . Bacterial mRNAs have a half-life measured in seconds to minutes; therefore, detection of bacteria-specific mRNAs would be evidence that metabolically active organisms are present . DESIGN: Blinded comparative study . PATIENTS: A total of 93 effusions from pediatric outpatients seen for myringotomy and tube placement for chronic (>3 months) OME (median age of children, 17 months) . SETTING: Tertiary care pediatric hospital . MAIN OUTCOME MEASURES: Percentage of positive test results for RT-PCR-based assays compared with culture for Haemophilus influenzae and concordance between RT-PCR and PCR-based findings for bacterial nucleic acids . RESULTS: Eleven (11.8%) of the 93 specimens tested positive by culture, PCR, and RT-PCR for H influenzae . A total of 29 specimens (31.2%) were positive by PCR but negative by culture for H influenzae . All 29 specimens were positive by RT-PCR for H influenzae-specific mRNA . CONCLUSIONS: The RT-PCR-based assay system can detect the presence of bacterial mRNA in a significant percentage of culturally sterile middle ear effusions, establishing the presence of viable, metabolically active, intact organisms in some culture-negative OME.

MMWR Morb Mortal Wkly Rep, 1998 Jan 16, 47(1), 8 - 12
Recommended childhood immunization schedule--United States, 1998; Efficacy and safety of a 10-day course of 400 or 600 milligrams of grepafloxacin once daily for treatment of acute bacterial exacerbations of chronic bronchitis: comparison with a 10-day course of 500 milligrams of ciprofloxacin twice daily; Veterans Affairs Outpatient Clinic, Boston, Massachusetts 02114, USAA randomized, prospective, double-blind, double-dummy, multicenter study investigated the efficacy and safety of 10 days of oral therapy with grepafloxacin at 400 mg once daily, grepafloxacin at 600 mg once daily, or ciprofloxacin at 500 mg twice daily in 624 patients with acute bacterial exacerbations of chronic bronchitis . At the end of treatment, clinical success (cure or improvement) was achieved for 93% (140 of 151), 88% (137 of 156), and 91% (145 of 160) of patients in the groups receiving grepafloxacin at 400 mg, grepafloxacin at 600 mg, and ciprofloxacin, respectively (clinically evaluable population) . At follow-up (14 to 28 days posttreatment), the clinical success rates were 87% (124 of 143), 81% (122 of 151), and 80% (123 of 154) in the groups receiving grepafloxacin at 400 mg and 600 mg and ciprofloxacin, respectively . A total of 379 pathogens were isolated from 290 patients, with the most common isolates being Moraxella catarrhalis (21%), Staphylococcus aureus (20%), Haemophilus influenzae (18%), and Streptococcus pneumoniae (7%) . For the evaluable population, successful bacteriologic response was obtained at the end of treatment for 96% (92 of 96), 98% (87 of 89), and 92% (82 of 90) of patients receiving grepafloxacin at 400 mg, grepafloxacin at 600 mg, and ciprofloxacin, respectively, and was maintained in 86% (82 of 95), 88% (78 of 89), and 82% (69 of 84) of patients, respectively, at follow-up . All pretreatment S . pneumoniae isolates were susceptible to grepafloxacin, but two strains were resistant to ciprofloxacin . All treatments were well tolerated, with the most frequently reported drug-related adverse events being nausea, taste perversion, and headache . All drug-related adverse events in the grepafloxacin groups were mild or moderate in severity . This study demonstrates that 10-day courses of grepafloxacin given at 400 or 600 mg once daily were as effective, clinically and bacteriologically, as ciprofloxacin given at 500 mg twice daily for the treatment of acute bacterial exacerbations of chronic bronchitis.

Acta Paediatr Jpn, 1997 Dec, 39(6), 676 - 80
Immunogenicity and safety of Haemophilus influenzae type b capsular polysaccharide tetanus conjugate vaccine (PRP-T) presented in a dual-chamber syringe with DTP; Kanra G et al.; Separate injections of Haemophilus influenzae type b capsular polysaccharide-tetanus conjugate (PRP-T) vaccine and diphtheria-tetanus-pertussis (DTP) reconstitution of freeze-dried PRP-T vaccine with liquid DTP vaccine have been shown to be safe and immunogenic in infants . The present study was conducted to test the safety and immunogenicity of the liquid combination vaccine administered to young infants in the dual-chamber syringe . The study was a monocenter, open clinical trial of 3 month-old infants receiving PRP-T and DTP vaccines in the dual-chamber syringe reconstituted prior to injection . Healthy infants were immunized according to a 3, 4 and 5 months-of-age schedule . The vaccine was administered in a dual-chamber syringe, ready to use with two chambers . The proximal chamber contained freeze-dried PRP-T and the distal chamber contained liquid combination-vaccine DTP . The freeze-dried PRP-T vaccine was reconstituted with the liquid DTP vaccine in the same unidose dual-chamber syringe (0.5 mL) and was injected intramuscularly into the deltoid region . Blood sampling was performed prior to vaccination at 3 months of age and after the third vaccination at 6 months . The primary end-point was the serological response to PRP-T vaccine as expressed by the percentage of infants with an antibody titer greater than or equal to 1 microgram/mL . The reactogenicity was expressed as the percentage of reported local and systemic reactions . A total of 108 infants were included in the study and received the dual-chamber syringe vaccine . After the third injection, all the infants had a PRP antibody titer greater than or equal to 0.15 microgram/mL and 94.4% of infants had a PRP antibody titer greater than or equal to 1 microgram/mL; the pertussis agglutinin titers were over the threshold 40 and 80 in all infants and 98.1% were over the threshold 320 . After the third injection, all the infants had diphtheria antibody titers greater than 0.1 IU/mL and 83.3% had titers greater than 1 IU/mL; all the infants had tetanus antibody titers greater than 0.1 IU/mL and 97.2% had results over 1 IU/mL . Thirty-seven infants (34.6%) had local reactions and 64.5% had systemic reactions . The dual-chamber syringe may reduce the cost of vaccine delivery, as well as the workload, and increase the vaccine acceptability and coverage rate of vaccines.

An Med Interna, 1997 Nov, 14(11), 554 - 8
{Moraxella (Branhamella) catarrhalis: its isolation in the respiratory secretions of adult patients}; Ferrer Marcelles A et al.; OBJECTIVE: We have designed a retrospective study in order to know the clinical significance of the isolation of Moraxella (Branhamella) catarrhalis (MC) in respiratory specimens of adult hospitalized patients . METHODS: We performed a Gram stain and culture on blood-agar, MacConkey media and quantitative culture in chocolate-agar to all respiratory samples . In patients with a clinical diagnosis of pneumonia BCYE-alpha was added . During 2 years (1992-1993) MC was isolated in respiratory specimens from 52 patients . We revised the clinical history of all these patients . RESULTS: MC was isolated in 60 respiratory specimens (sputum and/or tracheobronchial aspirates) from 52 patients . The Gram stain showed gram-negative cocci in 77% and gram-positive cocci in 17% of the cases . MC grew in pure culture in 28 specimens (46.6%) . In 23% of cases MC was isolated with Streptococcus pneumoniae and in 21% with Haemophilus influenzae . Fifty-two stocks (86.6%) produced beta-lactamase . Twelve patients had a clinical diagnosis of pneumonia, 8 of them had an underlying chronic respiratory disease . Other 24 patients with an underlying chronic respiratory disease had a bronchial infection as a cause of exacerbation of their respiratory disease . Seven patients without an underlying chronic respiratory disease had a clinical episode of acute bronchitis . Finally, in 9 patients the isolation of MC was considered a colonization . CONCLUSIONS: In 17% cases MC was identified as a gram-positive cocci in the Gram stain, which may cause false diagnosis . The etiological importance of MC in episodes of acute exacerbation of patients with an underlying chronic respiratory disease is high.

Pediatrics, 1998 Feb, 101(2), 242 - 9
Modulation of the immune system by human milk and infant formula containing nucleotides; Pickering LK et al.; OBJECTIVE: To determine whether human milk and nucleotides added to infant formula at levels present in human milk enhance development of the immune system during infancy . METHODS: A 12-month, controlled, randomized and blinded, multisite feeding trial was conducted on two infant formulas: iron-fortified, milk-based control formula (Control) or the same formula fortified with nucleotides (Nucleotide) . The level (72 mg/L) and ratio of individual nucleotides selected were patterned after those available in human milk . A third group fed human milk exclusively for 2 months and then human milk or Similac with iron until 12 months of age also was studied . Response to immunizations was chosen to assess development of the immune system . Infants followed the immunization schedule recommended by the American Academy of Pediatrics in 1991 . OUTCOME VARIABLES: Antibody responses were determined at 6, 7, and 12 months of age to Haemophilus influenzae type b polysaccharide (Hib), to diphtheria and tetanus toxoids, and to oral polio virus (OPV) immunizations . RESULTS: Of 370 full-term, healthy infants enrolled, 311 completed the study (107 Control, 101 Nucleotide, 103 human milk/Similac with iron) . Intake, tolerance, and growth of infants were similar in all three groups . Compared with the Control group 1 month after the third immunization (7 months of age), the Nucleotide group had a significantly higher Hib antibody concentration (geometric mean concentrations of 7.24 vs 4.05 micrograms/mL, respectively), and a significantly higher diphtheria antibody concentration (geometric mean of 1.77 vs 1.38 U/mL) . The significantly higher Hib antibody response in the Nucleotide group persisted at 12 months . The antibody responses to tetanus and OPV were not enhanced by nucleotide fortification . There also was an effect of breastfeeding on immune response . Infants who breastfed had significantly higher neutralizing antibody titers to polio virus than either formula-fed group (1:346 vs 1:169 and 1:192 in the Control and Nucleotide groups, respectively) at 6 months of age . CONCLUSION: Infant formula fortified with nucleotides enhanced H influenzae type b and diphtheria humoral antibody responses . Feeding human milk enhanced antibody responses to OPV . Dietary factors play a role in the antibody response of infants to immunization.

Ann Otol Rhinol Laryngol, 1998 Jan, 107(1), 10 - 6
Analysis of adult otitis media: polymerase chain reaction versus culture for bacteria and viruses; Liederman EM et al.; Recent studies using the polymerase chain reaction (PCR) have identified bacterial and viral genomic sequences in culture-negative pediatric middle ear effusions . To evaluate this technique in adults, 19 effusions were analyzed to compare bacterial and viral culture and PCR detection of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, and adenovirus . Effusions from 4 subjects positive for human immunodeficiency virus (HIV) were analyzed by PCR for HIV virus . Three of 19 effusions were culture-positive for bacteria, and 0 of 19 for viruses . Fifteen of 19 effusions were PCR-positive for bacterial genomic sequences, and 0 of 19 for adenovirus . Thirteen of 15 PCR-positive specimens demonstrated S pneumoniae, 5 of 15 H influenzae, and 0 of 13 M catarrhalis . All 4 effusions from HIV-positive subjects were PCR-positive for HIV . No effusion was culture-positive and PCR-negative . These results confirm that culture-negative middle ear effusions contain genomic sequences from bacterial pathogens . Finding of HIV RNA and DNA in effusion from HIV-positives suggests replicating virus in this fluid.

Scand J Infect Dis, 1997, 29(5), 485 - 9
Influence of age, gender and smoking on Streptococcus pneumoniae, Haemophilus influenzae and Moraxella (Branhamella) catarrhalis antibody titres in an elderly population; Kurtti P et al.; The purpose of this study was to investigate the association of serum antibody levels to Streptococcus pneumoniae, Haemophilus influenzae and Moraxella (Branhamella) catarrhalis with age, gender and smoking in an elderly population . The study population comprised all the inhabitants aged 65 years or over in a rural municipality in south-western Finland . Serum samples were obtained from 1,174 out of a total of 1,360 subjects . Bacterial antibodies were measured by enzyme immunoassay (EIA) using pneumolysin and whole bacterial cells of H . influenzae and M . catarrhalis (mixture of 10 different strains for both) as antigens . The main findings were as follows: (i) antibody levels generally decreased with increasing age both in men and in women; (ii) antibody titres against H . influenzae and M . catarrhalis were higher in men than in women; and (iii) antibody titres to H . influenzae and M . catarrhalis, but not to S . pneumoniae, were significantly higher in smokers than in non-smokers . These data suggest that antibody-mediated protection against respiratory pathogens may be impaired in the elderly, leading to a higher susceptibility to respiratory tract infections, that the exposure to H . influenzae and M . catarrhalis may be higher in men than in women, and that smokers have more respiratory infections or colonization due to these 2 bacteria than do non-smokers.

Planta Med, 1997 Dec, 63(6), 508 - 10
Antibacterial activity of extracts and constituents of Pelargonium sidoides and Pelargonium reniforme; Kayser O et al.; The antibacterial activity of extracts and isolated constituents (scopoletin, umckalin, 5,6,7-trimethoxycoumarin, 6,8-dihydroxy-5,7-dimethoxycoumarin, (+)-catechin, gallic acid and its methyl ester) of Pelargonium sidoides and Pelargonium reniforme (Geraniaceae), plant species used in folk medicine by the Southern African native population, was evaluated against 8 microorganisms, including 3 Gram-positive (Staphylococcus aureus, Streptococcus pneumoniae, and beta-hemolytic Streptococcus 1451) and 5 Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Haemophilus influenzae) . Minimum inhibitory concentrations (MICs) varied with the preparation of the extracts and microorganisms tested, from about 0.6 mg/ml for aqueous phases to over 10 mg/ml for crude Pelargonium extracts . With the exception of the ineffective (+)-catechin, all the potentially active compounds exhibited antibacterial activities with MICs of 200-1000 micrograms/ml . The results provide for a rational basis of the traditional use of the titled Pelargonium species.

Gene, 1997 Dec 19, 204(1-2), 185 - 94
Creation of an isogenic P1-deficient mutant of Haemophilus influenzae biogroup aegyptius; Segada LM et al.; Haemophilus influenzae biogroup aegyptius, the causative agent of Brazilian purpuric fever (BPF), expresses a heat-modifiable 48 kDa outer membrane protein, P1, which is conserved in most Brazilian case-clone isolates . To study the role of P1 in pathogenesis of BPF we constructed via homologous recombination an isogenic P1-deficient mutant of H . influenzae biogroup aegyptius . The procedure involved a modification of Hererot's method for development of competence . Modifications included variations in the growth conditions, use of cAMP, specific characteristics of the donor DNA, and antibiotic selection . P1-deficient mutants were confirmed by SDS-PAGE, loss of reactivity with a specific monoclonal antibody on Western blot, restriction analysis and Southern blot . Our results establish the first successful transformation of homologous DNA into H . influenzae biogroup aegyptius.

Res Vet Sci, 1997 Sep-Oct, 63(2), 145 - 9
Evaluation of a competitive immunoassay for the detection of bovine haptoglobin; McNair J et al.; A competitive immunoassay to quantify the serum concentration of bovine haptoglobin (hp) using time resolved fluorescence was compared with an indirect method of hp assay (haemoglobin binding assay), using sera taken from healthy animals (n = 158), animals experimentally infected with Haemophilus somnus (n = 10) and from sick animals requiring veterinary treatment (n = 440) . Upper limits of normality (for normal animals) were tentatively established for the immunoassay (2.1 micrograms ml-1) and for the haemoglobin binding assay (103 micrograms ml-1) . The competitive immunoassay detected elevated hp in 62.5 per cent of field sera by comparison with only 19.2 per cent using the conventional haemoglobin binding assay . Serum albumin concentration did not correlate with hp although concentrations of globulins and copper did correlate . However, these parameters (serum globulin and copper) were found to be insensitive markers of inflammatory disease.

J Pediatr, 1997 Dec, 131(6), 869 - 73
Effect of nationwide vaccination of 3-month-old infants in The Netherlands with conjugate Haemophilus influenzae type b vaccine: high efficacy and lack of herd immunity; van Alphen L et al.; OBJECTIVE: The effect of nationwide vaccination in The Netherlands with conjugate Haemophilus influenzae type b vaccine on the incidence of H . influenzae meningitis was assessed in the first 3 years after the introduction of vaccination to the birth cohort at 3 months of age . STUDY DESIGN: Children in The Netherlands born after April 1, 1993, were vaccinated at the age of 3, 4, 5, and 11 months . Children older than 3 months at the inception of the H . influenzae type b vaccination program were not immunized . The number of cases among the vaccine cohort was compared with the number in a historical control group of children born in the period April 1, 1990, and April 1, 1993 . RESULTS: The total number of patients with meningitis caused by H . influenzae type b reached a low, but constant level, that was expected for absence of herd immunity . Among children in the vaccine era group 22 cases occurred, whereas among the historical control group (prevaccine era) 342 cases were found . In the vaccine era cohort, 2 patients with H . influenzae type b meningitis had been vaccinated three times, 13 received one or no vaccine dose because of their age, and 7 were not vaccinated for religious or logistic reasons . The number of cases among nonvaccinated children older than 3 years and the number of H . influenzae meningitis cases caused by strains other than type b did not change . CONCLUSIONS: Conjugate H . influenzae type b vaccine prevents H . influenzae type b meningitis very effectively (99.4%) in children vaccinated twice or more . To reach rapid prevention of all H . influenzae type b disease simultaneous introduction of H . influenzae type b vaccination of children at various ages is recommended.

Nat Med, 1998 Jan, 4(1), 88 - 91
Enhancement of T cell-independent immune responses in vivo by CD40 antibodies; Dullforce P et al.; In this report we describe a potentially powerful method for vaccinating infants against encapsulated bacterial pathogens such as Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis . High levels of antibody directed against the polysaccharides of the bacterial capsule are normally protective . Unfortunately, the capsular polysaccharides are T cell-independent antigens (TI); lacking T-cell help, they induce only weak, predominantly IgM antibody responses, with infants responding especially poorly . T-cell help, given to B cells during responses to protein antigens, causes stronger antibody responses and isotype switching to the IgG isotypes . T-cell help is mainly mediated through ligation of the B-cell surface antigen, CD40, by its cognate T-cell ligand, CD154 . Here we show that administering anti-CD40 monoclonal antibody to mice, along with pneumococcal polysaccharide, provides a substitute for T-cell help and results in the generation of strong, isotype-switched antibody responses, which are protective . The work points the way toward a possible effective and inexpensive means of protecting susceptible groups against important bacterial pathogens.

Pediatr Infect Dis J, 1997 Dec, 16(12), 1135 - 40
A glycoprotein pneumococcal conjugate vaccine primes for antibody responses to a pneumococcal polysaccharide vaccine in Gambian children; Obaro SK et al.; BACKGROUND: Streptococcus pneumoniae is a major cause of acute respiratory infections and acute bacterial meningitis in children . Pneumococcal polysaccharide vaccines are poorly immunogenic in this highly vulnerable group, but protein polysaccharide conjugate vaccines are likely to be more effective . OBJECTIVES: To determine whether immunization of infants with a pneumococcal conjugate vaccine induces immunologic memory . METHODS: Eighty-four Gambian children, who had been vaccinated previously with two or three doses of a pentavalent pneumococcal conjugate vaccine (CRM197) or with a Haemophilus influenzae type b (Hib) conjugate vaccine were immunized when approximately 2 years old with a 23-valent pneumococcal polysaccharide vaccine, and a blood sample was obtained 10 days later . Pneumococcal antibody titers in prevaccination and postvaccination sera were measured by enzyme-linked immunosorbent assay and by an opsonophagocytic assay . RESULTS: On revaccination with a pneumococcal polysaccharide vaccine, children who had previously received pneumococcal conjugate vaccine had higher antibody concentrations to each of the five polysaccharide components of the conjugate vaccine than did control children . For type 6B polysaccharide, which is poorly immunogenic in young children, postvaccination antibody concentrations were 0.37, 27.6 and 50.9 microg/ml in children who had received no previous pneumococcal immunization or two or three doses of conjugate vaccine, respectively . Type 14 antibodies produced after revaccination were of high avidity and had opsonic activity . CONCLUSION: Vaccination of young infants with two or three doses of a pneumococcal conjugate vaccine primes the immune system to respond strongly and rapidly on subsequent exposure to pneumococcal polysaccharide.

Pediatr Infect Dis J, 1997 Dec, 16(12), 1122 - 30
Haemophilus influenzae type b-specific antibody in infants after maternal immunization; Englund JA et al.; OBJECTIVE: To study the kinetics of Haemophilus influenzae type b (Hib)-specific antibody in infants born to mothers immunized with an Hib polysaccharide or one of two Hib conjugate vaccines . STUDY DESIGN: Serum antibody to the polyribosylribitol (PRP) moiety of Hib was measured by radioimmunoassay and enzyme-linked immunosorbent assay at birth and at 2 and 6 months of age in infants born to women immunized with Hib polysaccharide or conjugate vaccine (PRP-D and HbOC) . A subset of infants > or = 6 months of age was immunized with Hib conjugate vaccine after licensure of this vaccine for infants . A comparison group of 18 infants born to unimmunized women received the same Hib conjugate vaccine on a similar schedule . RESULTS: Total PRP antibody concentrations were 1.50, 14.4 and 20.4 microg/ml in 2-month-old infants born to mothers immunized with polysaccharide, PRP-D and HbOC vaccines, respectively, and 2.54, 1.35 and 2.46 microg/ml in 6-month-old infants . Infants born to mothers immunized with polysaccharide vaccine had significantly less PRP antibody at 2 months of age but similar antibody concentrations at 6 months of age . Persistence or increases in total PRP antibody during 6 months were noted in 21 of 47 (44.6%) study infants . A subset of study and comparison infants was immunized with a mean of 2.6 doses of Hib vaccines between 6 months and 2 years of age, and all infants had total PRP antibody concentrations > or = 0.15 microg/ml . CONCLUSION: Conjugate Hib vaccines administered during the last trimester of pregnancy resulted in significantly higher PRP antibody titers in infants at birth and 2 months of age than did polysaccharide vaccine . A subset of infants born to immunized mothers was subsequently immunized with Hib conjugate vaccine and had antibody concentrations similar to those in infants born to nonimmunized women.

Pediatr Infect Dis J, 1997 Dec, 16(12), 1113 - 21
Safety and immunogenicity of a combined pentavalent diphtheria, tetanus, acellular pertussis, inactivated poliovirus and Haemophilus influenzae type b-tetanus conjugate vaccine in infants, compared with a whole cell pertussis pentavalent vaccine; Dagan R et al.; BACKGROUND: We compared the safety and immunogenicity of two combined diphtheria-tetanus-pertussis-inactivated poliovirus vaccines containing either acellular (Pa, SmithKline Beecham Biologicals) or whole cell (Pw, Pasteur Merieux Connaught) pertussis components, mixed with a Haemophilus influenzae type b polysaccharide polyribosylribitol phosphate-tetanus conjugate vaccine in an open, randomized study in healthy infants . DESIGN: The combined vaccines were given at 2, 4, 6 and 12 months of age, and serum samples were obtained at ages 2, 6, 7, 12 and 13 months . Adverse events were obtained by diary cards . RESULTS: The Pa group (n = 101) had a clearly lower incidence of both local and systemic adverse events than the Pw group (n = 100) . Immunogenicity was comparable for the diphtheria and tetanus components, but significantly superior for pertussis toxin, filamentous hemagglutinin, pertactin and polioviruses 1, 2 and 3 in the Pa group . Both groups had an appropriate response with regard to H . influenzae type b polysaccharide polyribosylribitol phosphate, but the dynamics of the response were significantly different: geometric mean concentrations (micrograms per ml) after the second, third and booster doses were 1.27, 5.06 and 23.12 in the Pa group and 2.72, 6.66 and 13.59 in the Pw group, respectively (P = 0.0002 after second dose; P = 0.0005 after booster) . CONCLUSION: The presently studied diphtheria, tetanus, acellular pertussis-H . influenzae b vaccine conjugated to tetanus toxoid combination was at least as immunogenic as the diphtheria, tetanus, whole cell pertussis-H . influenzae b vaccine conjugated to tetanus toxoid combination, with a significantly better safety profile . This is of obvious importance in countries where inactivated poliovirus vaccine is part of the routine infant immunization programs.

Infect Immun, 1998 Jan, 66(1), 364 - 8
Prevalence and distribution of the hmw and hia genes and the HMW and Hia adhesins among genetically diverse strains of nontypeable Haemophilus influenzae; St Geme JW 3rd et al.; Nontypeable Haemophilus influenzae is a common cause of human disease and initiates infection by colonizing the upper respiratory tract . In previous work we identified high-molecular-weight adhesins referred to as HMW1 and HMW2, expressed by nontypeable strain 12, and determined that most strains of nontypeable H . influenzae express one or two antigenically related proteins . More recently, we determined that some strains lack HMW1- and HMW2-like proteins and instead express an adhesin called Hia . In the present study, we determined the prevalence and distribution of the hmw and hia genes in a collection of 59 nontypeable strains previously characterized in terms of genetic relatedness . Based on Southern analysis, 47 strains contained sequences homologous to the hmw1 and hmw2 genes and nine strains contained homologs to hia . No strain harbored both hmw and hia, and three strains harbored neither . Although the hmw and hia genes failed to define distinct genetic divisions, the hmw-deficient strains formed small clusters or lineages within the larger population structure . Additional analysis established that the IS1016 insertion element was uniformly absent from strains containing hmw sequences but was present in two-thirds of the hmw-deficient strains . As IS1016 is associated with the capsule locus (cap) in most encapsulated strains of H . influenzae, we speculate that hmw-deficient nontypeable strains evolved more recently from an encapsulated ancestor.

Infect Immun, 1998 Jan, 66(1), 151 - 60
Role of the Haemophilus ducreyi Ton system in internalization of heme from hemoglobin; Elkins C et al.; By cloning into Escherichia coli and construction of isogenic mutants of Haemophilus ducreyi, we showed that the hemoglobin receptor (HgbA) is TonB dependent . An E . coli hemA tonB mutant expressing H . ducreyi hgbA grew on low levels of hemoglobin as a source of heme only when an intact H . ducreyi Ton system plasmid was present . In contrast, growth on heme by the E . coli hemA tonB mutant expressing hgbA was observed only at high concentrations of heme, was TonB independent, and demonstrated that H . ducreyi HgbA was not sufficient to function as a typical TonB-dependent heme receptor in E . coli . Allelic replacement of the wild-type H . ducreyi exbB, exbD, and tonB loci with the exbB, exbD, and tonB deletion resulted in an H . ducreyi isogenic mutant unable to utilize hemoglobin but able to utilize hemin at the same levels as the parent strain to fulfill its heme requirement . This finding confirms the TonB dependence of HgbA-mediated hemoglobin utilization and suggests that uptake of hemin in H . ducreyi is TonB independent . Additionally, the H . ducreyi Ton system mutant synthesized increased amounts of HgbA and other heme-regulated outer membrane proteins, consistent with derepression of these proteins due to lower intracellular heme and/or iron concentrations in the mutant . Sequencing of the Ton system genes revealed that the arrangement of the genes was exbB exbD tonB . The proximity and structure of these genes suggested that they are transcribed as an operon . This arrangement, as well as the DNA and deduced amino acid sequences of these H . ducreyi genes, was most similar to those from other pasteurellae.

J Bacteriol, 1998 Jan, 180(1), 107 - 18
The tryptophanase gene cluster of Haemophilus influenzae type b: evidence for horizontal gene transfer; Martin K et al.; Among strains of Haemophilus influenzae, the ability to catabolize tryptophan (as detected by indole production) varies and is correlated with pathogenicity . Tryptophan catabolism is widespread (70 to 75%) among harmless respiratory isolates but is nearly universal (94 to 100%) among strains causing serious disease, including meningitis . As a first step in investigating the relationship between tryptophan catabolism and virulence, we have identified genes in pathogenic H . influenzae which are homologous to the tryptophanase (tna) operon of Escherichia coli . The tna genes are located on a 3.1-kb fragment between nlpD and mutS in the H . influenzae type b (Eagan) genome, are flanked by 43-bp direct repeats of an uptake signal sequence downstream from nlpD, and appear to have been inserted as a mobile unit within this sequence . The organization of this insertion is reminiscent of pathogenicity islands . The tna cluster is found at the same map location in all indole-positive strains of H . influenzae surveyed and is absent from reference type d and e genomes . In contrast to H . influenzae, most other Haemophilus species lack tna genes . Phylogenetic comparisons suggest that the tna cluster was acquired by intergeneric lateral transfer, either by H . influenzae or a recent ancestor, and that E . coli may have acquired its tnaA gene from a related source . Genomes of virulent H . influenzae resemble those of pathogenic enterics in having an island of laterally transferred DNA next to mutS.

J Med Liban, 1997, 45(1), 40 - 2
Post immunization Hib antigen detection in the CSF of a patient with meningococcal meningitis; Samaha AN et al.; We report a case of meningococcal meningitis where the cerebrospinal fluid was negative for Neisseria meningitidis but positive for Haemophilus influenzae type b by rapid antigen detection test . We believe that this was due to prior immunization with Haemophilus influenzae type b vaccine . We recommend caution in interpretation of the rapid antigen detection tests especially in patients who had been vaccinated against organisms screened by these tests.

Electrophoresis, 1997 Oct, 18(11), 2085 - 90
Effect of protein application mode and acrylamide concentration on the resolution of protein spots separated by two-dimensional gel electrophoresis; Langen H et al.; Two-dimensional gel electrophoresis separates large numbers of proteins in two steps on the basis of differences in their pIs and molecular masses . The separation is usually performed on immobilized pH gradient strips, followed by gradient polyacrylamide gels separating proteins with molecular masses between 5-200 kDa . For the first-dimensional separation the protein samples are usually applied near one end of the strip . Using total soluble protein extracts of the bacterium Haemophilus influenzae, we found that simultaneous sample application at both the basic and the acidic ends of the strip resulted in detection of more and stronger protein spots in comparison with sample application at one end only . Because many proteins of an organism have similar pI and Mr values, an overlapping of protein spots is frequently observed in the second-dimensional separation . The soluble protein fraction of H . influenzae was further separated on gels of constant acrylamide concentration between 7.5% and 15.0% . We found that for proteins of molecular mass within certain ranges, the gels of homogeneous acrylamide concentration provided more efficient spot separation than the gradient gels . The observed improvements in spot resolution may be useful in the characterization of proteins from other organisms or cell lines.

J Mol Biol, 1997 Dec 12, 274(4), 562 - 76
A structural census of genomes: comparing bacterial, eukaryotic, and archaeal genomes in terms of protein structure; Gerstein M; Representative genomes from each of the three kingdoms of life are compared in terms of protein structure, in particular, those of Haemophilus influenzae (a bacteria), Methanococcus jannaschii (an archaeon), and yeast (a eukaryote) . The comparison is in the form of a census (or comprehensive accounting) of the relative occurrence of secondary and tertiary structures in the genomes, which particular emphasis on patterns of supersecondary structure . Comparison of secondary structure shows that the three genomes have nearly the same overall secondary-structure content, although they differ markedly in amino acid composition . Comparison of super-secondary structure, using a novel "frequent-words" approach, shows that yeast has a preponderance of consecutive strands (e.g . beta-beta-beta patterns), Haemophilus, consecutive helices (alpha-alpha-alpha), and Methanococcus, alternating helix-strand structures (beta-alpha-beta) . Yeast also has significantly more helical membrane proteins than the other two genomes, with most of the differences concentrated in proteins containing two transmembrane segments . Comparison of tertiary structure (by sequence matching and domain-level clustering) highlights the substantial duplication in each genome (approximately 30% to 50%), with the degree of duplication following similar patterns in all three . Many sequence families are shared among the genomes, with the degree of overlap between any two genomes being roughly similar . In total, the three genomes contain 148 of the approximately 300 known protein folds . Forty-five of these 148 that are present in all three genomes are especially enriched in mixed super-secondary structures (alpha/beta) . Moreover, the five most common of these 45 (the "top-5") have a remarkably similar super-secondary structure architecture, containing a central sheet of parallel strands with helices packed onto at least one face and beta-alpha-beta connections between adjacent strands . These most basic molecular parts, which, presumably, were present in the last common ancestor to the three Kingdoms, include the TIM-barrel, Rossmann, flavodoxin, thiamin-binding, and P-loop-hydrolase folds.

Genomics, 1997 Dec 1, 46(2), 200 - 16
Analysis of protein domain families in Caenorhabditis elegans; Sonnhammer EL et al.; The Caenorhabditis elegans genome sequencing project has completed over half of this nematode's 100-Mb genome . Proteins predicted in the finished sequence have been compiled and released in the data-base Wormpep . Presented here is a comprehensive analysis of protein domain families in Wormpep 11, which comprises 7299 proteins . The relative abundance of common protein domain families was counted by comparing all Wormpep proteins to the Pfam collection of protein families, which is based on recognition by hidden Markov models . This analysis also identified a number of previously unannotated domains . To investigate new apparently nematode-specific protein families, Wormpep was clustered into domain families on the basis of sequence similarity using the Domainer program . The largest clusters that lacked clear homology to proteins outside Nematoda were analyzed in further detail, after which some could be assigned a putative function . We compared all proteins in Wormpep 11 to proteins in the human, Saccharomyces cerevisiae, and Haemophilus influenzae genomes . Among the results are the estimation that over two-thirds of the currently known human proteins are likely to have a homologue in the whole C . elegans genome and that a significant number of proteins are well conserved between C . elegans and H . influenzae, that are not found in S . cerevisiae.

Protein Sci, 1997 Dec, 6(12), 2525 - 37
A flavodoxin that is required for enzyme activation: the structure of oxidized flavodoxin from Escherichia coli at 1.8 A resolution; Hoover DM et al.; In Escherichia coli, flavodoxin is the physiological electron donor for the reductive activation of the enzymes pyruvate formate-lyase, anaerobic ribonucleotide reductase, and B12-dependent methionine synthase . As a basis for studies of the interactions of flavodoxin with methionine synthase, crystal structures of orthorhombic and trigonal forms of oxidized recombinant flavodoxin from E . coli have been determined . The orthorhombic form (space group P2(1)2(1)2(1), a = 126.4, b = 41.10, c = 69.15 A, with two molecules per asymmetric unit) was solved initially by molecular replacement at a resolution of 3.0 A, using coordinates from the structure of the flavodoxin from Synechococcus PCC 7942 (Anacystis nidulans) . Data extending to 1.8-A resolution were collected at 140 K and the structure was refined to an Rwork of 0.196 and an Rfree of 0.250 for reflections with I > 0 . The final model contains 3,224 non-hydrogen atoms per asymmetric unit, including 62 flavin mononucleotide (FMN) atoms, 354 water molecules, four calcium ions, four sodium ions, two chloride ions, and two Bis-Tris buffer molecules . The structure of the protein in the trigonal form (space group P312, a = 78.83, c = 52.07 A) was solved by molecular replacement using the coordinates from the orthorhombic structure, and was refined with all data from 10.0 to 2.6 A (R = 0.191; Rfree = 0.249) . The sequence Tyr 58-Tyr 59, in a bend near the FMN, has so far been found only in the flavodoxins from E . coli and Haemophilus influenzae, and may be important in interactions of flavodoxin with its partners in activation reactions . The tyrosine residues in this bend are influenced by intermolecular contacts and adopt different orientations in the two crystal forms . Structural comparisons with flavodoxins from Synechococcus PCC 7942 and Anaebaena PCC 7120 suggest other residues that may also be critical for recognition by methionine synthase.

Clin Ther, 1997 Sep-Oct, 19(5), 989 - 1001
Efficacy of ciprofloxacin and clarithromycin in acute bacterial exacerbations of complicated chronic bronchitis: interim analysis . Bronchitis Study Group; Anzueto A et al.; In a multicenter, community-based study involving more than 300 primary care physicians in the United States, the efficacy and safety of ciprofloxacin and clarithromycin were compared in the treatment of patients with complicated or severe acute bacterial exacerbations of chronic bronchitis (i.e., those who had failed previous antibiotic treatment within the prior 2 to 4 weeks; those with susceptibility data suggestive of a resistant pathogen; those having three or more acute exacerbations of chronic bronchitis {AECB} within the past year; and those having three or more comorbid conditions) . Patients were randomized to either ciprofloxacin (CIP) 750 mg BID or clarithromycin (CLR) 500 mg BID, both administered for 10 days; all patients were treated on an outpatient basis . Clinical response at the end of therapy was the primary efficacy variable . An interim analysis was performed on the results from 743 patients (369 CIP, 374 CLR) with clinical and bacteriologic evidence of a bronchopulmonary infection who had completed an ongoing study as of the end of May 1997 . Three hundred nine pathogens were isolated before therapy, including Haemophilus spp (75 isolates), Moraxella catarrhalis (67 isolates), Staphylococcus aureus (55 isolates), and Streptococcus pneumoniae (23 isolates) . Seven hundred eighteen patients (97%) were included in the efficacy-valid population . Clinical success at the end of therapy was observed in 90% (272 of 302) and 88% (274 of 313) of efficacy-valid patients treated with CIP and CLR, respectively (95% confidence interval {CI} = -2.4 to 7.6) . Corresponding rates for the intent-to-treat population were also 90% (283 of 314) and 88% (281 of 321), respectively (95% CI = -2.3 to 7.5) . The bacteriologic response for efficacy-valid patients at the end of therapy was 98% (119 of 122) for CIP-treated and 93% (103 of 111) for CLR-treated patients (95% CI = -0.8 to 10.2) . The eradication rates for the three most commonly isolated gram-negative pathogens were 100% for CIP-treated and 95% for CLR-treated patients and 96% each for the two most commonly isolated gram-positive organisms . Superinfections due to respiratory tract pathogens were more common in the CLR group (10 organisms) than in the CIP group (4 organisms) . Seventy-four (20%) CIP-treated and 62 (17%) CLR-treated patients reported 118 and 103 respective study-emergent adverse events . Headache, abdominal pain, diarrhea, nausea, and vomiting in CIP-treated patients and diarrhea, nausea, vomiting, and taste perversion in CLR-treated patients were the most commonly reported adverse events . Treatment of patients with complicated or severe AECB with CIP 750 mg BID was associated with rates of clinical success and bacteriologic eradication similar to those with CLR.

Clin Ther, 1997 Sep-Oct, 19(5), 975 - 88
Efficacy and safety of grepafloxacin 600 mg daily for 10 days in patients with community-acquired pneumonia; Topkis S et al.; The efficacy and safety of grepafloxacin in treating patients with community-acquired pneumonia (CAP) was assessed in an open-label, noncomparative study . Patients (N = 273) received grepafloxacin 600 mg QD for 10 days . A total of 237 patients (87%) completed the study . In assessable patients, the clinical success rate at follow-up (4 to 6 weeks after the last dose) was 89% (211/238 patients) . In microbiologically assessable patients, the eradication rate at follow-up was 95% (86/91 isolates) . Grepafloxacin was highly effective in the treatment of bacterial CAP caused by Streptococcus pneumoniae (irrespective of penicillin susceptibility), Haemophilus influenzae, Haemophilus parainfluenzae, Moraxella catarrhalis, and Staphylococcus aureus and in the therapy of atypical pneumonia caused by Mycoplasma pneumoniae and Legionella pneumophila . Grepafloxacin was well tolerated, with the most frequently reported drug-related adverse events being taste perversion and nausea . Grepafloxacin 600 mg QD for 10 days was highly effective and well tolerated in the treatment of patients with CAP.

Prep Biochem Biotechnol, 1997 Nov, 27(4), 253 - 69
Purification and characterization of a 31-kilodalton iron-regulated periplasmic protein from Pasteurella haemolytica A1; Tabatabai LB et al.; A prominent iron-regulated periplasmic protein was purified from Pasteurella haemolytica grown in an iron-deficient chemically defined medium . The protein was purified by anion exchange chromatography and appeared as a single band by SDS-PAGE with a molecular weight of 32,000 . A yield of five mg was obtained from 91 mg of protein extract . The iron-regulated protein existed as a monomer in the native state with an average molecular weight of 29,877 as determined by analytical ultracentrifugation . The protein had a molecular weight of 30,880 as determined by matrix-assisted laser desorption mass spectrometry, hence the protein is referred to as the 31 kDa protein . Isoelectric focusing showed four bands with pIs of 7.15, 6.8, 6.6, and 5.9 . The secondary structure of the protein was determined by circular dichroism and contained 16% alpha-helical structure . The N-terminal sequence, EPFKVVTTFTVIQDIAQNVAGDKAT, showed a 95% identity with the 31 kDa iron-binding protein from Haemophilus influenzae . Isolation and characterization of iron-regulated proteins are of particular interest because of their potential roles in iron assimilation and microbial virulence.

Antibiot Khimioter, 1997, 42(10), 29 - 32
{Efficacy and tolerance of coamoxiclav in community-acquired lower respiratory tract infections in children}; Blokhin BM et al.; Amoxyclav (amoxycillin/potassium clavulanate, A/PC) was used in the treatment of 55 children with acute bronchitis and pneumonia . The drug was administered in a dose of 20-40 mg/kg body weight a day in 3 portions . The treatment course was 4 to 10 days . The treatment was performed under careful clinicoroent-genologic control . The clinical picture of the disease in the children was characterized by a moderate process which made it possible to treat the children as outpatients . The clinical efficacy amounted to 90.5 per cent . The isolates of Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus and Haemophilus influenzae proved to be susceptible to A/PC . It may be used as the 1st class agent in the treatment of children with lower respiratory tract infection.

Przegl Epidemiol, 1997, 51(3), 303 - 7
{Etiologic factors for purulent meningitis in children . Twenty years of personal observations}; Patrzalek M et al.; Etiological agents in purulent meningitis cases hospitalized during two 9 years periods: 1977-1985 and 1988-1996 were compared . While the percentage of cases caused by Neisseria meningitidis was similar in the two analyzed periods (55.7% i 66.5%, respectively), the percentage of cases caused by Streptococcus pneumoniae decreased from 33.5% to 8.4% and the percentages of cases caused by Haemophilus influenzae type b increased from 0.6% to 14.2% . The analysis of the sensitivity of isolated strains of H . influenzae type b shows the occurrence of strains resistant to ampicillin and chloramphenicol.

Dtsch Tierarztl Wochenschr, 1997 Sep, 104(9), 374 - 8
{Bronchoalveolar lavage on pig breeding farms}; Delbeck F et al.; In 182 pigs lung lavage was performed using a endotracheal tube and a catheter . The collected bronchoalveolar lavage fluid (BALF) was examined microbiologically . With decreasing numbers alpha-hamolytic Streptococci, Bordetella bronchiseptica, Haemophilus parasuis, Pasteurella multocida were cultured . Actinobacillus pleuropneumoniae was isolated from 3 BALFs . In one farm piglets were lavaged routinely for monitoring of the lung health status.

J Antimicrob Chemother, 1997 Oct, 40(4), 579 - 82
Failure of Neisseria gonorrhoeae to grow in the ATB NH susceptibility test system; Young H et al.; The ATB NH system designed for antibiotic susceptibility testing of Neisseria gonorrhoeae, Neisseria meningitidis and Haemophilus influenzae was evaluated using 94 clinical isolates of gonococci representing a wide variety of serovar/auxotype strains . Using the manufacturer's automated system 55% of the clinical isolates failed to grow, compared with a 33% failure rate for manual processing and visual reading . Growth failure was significantly higher with 1A isolates (73% automated and 69% manual) than with 1B isolates (49% automated and 25% manual) . The higher failure rate of 1A isolates correlated with multiple auxotrophy . The inability of the ATB NH system to support the growth of common serovar/auxotypes makes the ATB NH system unsuitable for antibiotic susceptibility testing of N . gonorrhoeae.

J Antimicrob Chemother, 1997 Oct, 40(4), 573 - 7
Concentrations of levofloxacin (HR 355) in the respiratory tract following a single oral dose in patients undergoing fibre-optic bronchoscopy; Andrews JM et al.; Concentrations of levofloxacin were measured in bronchial biopsies, alveolar macrophages (AM), epithelial lining fluid (ELF) and serum following a single oral dose . Concentrations were measured by a microbiological assay method . A total of 35 patients undergoing fibre-optic bronchoscopy were studied . Mean serum, AM, ELF and biopsy concentrations were as follows . 0.5 h: 4.73 mg/L, 19.1 mg/L, 4.74 mg/L and 4.3 mg/kg; 1 h: 6.6 mg/L, 32.5 mg/L, 10.8 mg/L and 8.3 mg/kg; 2 h: 4.9 mg/L, 41.9 mg/L, 9.0 mg/L and 6.5 mg/kg; 4 h: 4.1 mg/L, 27.7 mg/L, 10.9 mg/L and 6.0 mg/kg; and 6-8 h: 4.0 mg/L, 38.4 mg/L, 9.6 mg/L and 4.0 mg/kg respectively . Mean serum and AM concentrations at 12-24 h were 1.2 and 13.9 mg/L respectively (concentrations in biopsy and ELF were only measurable in three of the six patients) . These concentrations exceed the MIC90s of the common respiratory pathogens, Haemophilus influenzae (0.015 mg/L), Moraxella catarrhalis (0.06 mg/L) and Streptococcus pneumoniae (1 mg/L) and suggest that levofloxacin should be efficacious in the treatment of community- and hospital-acquired respiratory infection.

Mol Microbiol, 1997 Nov, 26(3), 505 - 18
Structural determinants of processing and secretion of the Haemophilus influenzae hap protein; Hendrixson DR et al.; Haemophilus influenzae elaborates a surface protein called Hap, which is associated with the capacity for intimate interaction with cultured epithelial cells . Expression of hap results in the production of three protein species: outer membrane proteins of approximately 155 kDa and 45 kDa and an extracellular protein of approximately 110 kDa . The 155 kDa protein corresponds to full-length mature Hap (without the signal sequence), and the 110 kDa extracellular protein represents the N-terminal portion of mature Hap (designated Haps) . In the present study, we examined the mechanism of processing and secretion of Hap . Site-directed mutagenesis suggested that Hap is a serine protease that undergoes autoproteolytic cleavage to generate the 110 kDa extracellular protein and the 45 kDa outer membrane protein . Biochemical analysis confirmed this conclusion and established that cleavage occurs on the bacterial cell surface . Determination of N-terminal amino acid sequence and mutagenesis studies revealed that the 45 kDa protein corresponds to the C-terminal portion of Hap, starting at N1037 . Analysis of the secondary structure of this protein (designated Hap beta) predicted formation of a beta-barrel with an N-terminal transmembrane alpha-helix followed by 14 transmembrane beta-strands . Additional analysis revealed that the final beta-strand contains an amino acid motif common to other beta-barrel outer membrane proteins . Upon deletion of this entire C-terminal consensus motif, Hap could no longer be detected in the outer membrane, and secretion of Haps was abolished . Deletion or complete alteration of the final three amino acid residues had a similar but less dramatic effect, suggesting that this terminal tripeptide is particularly important for outer membrane localization and/or stability of the protein . In contrast, isolated point mutations that disrupted the amphipathic nature of the consensus motif or eliminated the C-terminal tryptophan had no effect on outer membrane localization of Hap or secretion of Haps . These results provide insight into a growing family of Gram-negative bacterial exoproteins that are secreted by an IgA1 protease-like mechanism; in addition, they contribute to a better understanding of the structural determinants of targeting of beta-barrel proteins to the bacterial outer membrane.

Clin Diagn Virol, 1997 Nov, 8(3), 209 - 17
Molecular epidemiology of rabies epizootics in Texas; Rohde RE et al.; BACKGROUND: Texas is in the midst of two independent epizootics of rabies, involving coyotes (Canis latrans) and domestic dogs (Canis familiaris) in southern Texas and grey foxes (Urocyon cinereoargenteus) in west central Texas . The domestic dog/coyote (DDC) and grey for (TF) rabies virus variants cannot be differentiated by antigenic typing with currently available monoclonal antibodies . These two variants also cannot be distinguished from a third variant, Sonora dog (SD) rabies, that is not enzootic in Texas, but occasionally occurs in animals along the western border with Mexico . OBJECTIVES: To determine a method for the differentiation of the DDC . TF and SD variants, which is essential for epidemiologic monitoring of the Oral Rabies Vaccination Program (ORVP), a program instituted to control rabies in coyotes and grey foxes in Texas . STUDY DESIGN: Primers complementary to nucleoprotein sequence of either the DDC or TF rabies virus permit specific reverse transcription and amplification by polymerase chain reaction . In addition, general primers, which recognize a broad range of rabies variants, used in conjunction with a restriction digest for the differentiation of DDC, TF of SD rabies virus were investigated . RESULTS AND CONCLUSIONS: Of 122 specimens tested with specific primers . 111 (91%) were specifically identified as either DDC (33 samples) or TF (78 samples) . Overly stringent conditions, enzyme inhibitors, or limiting RNA may account for the 11 non-amplifications . Amplification of RNA under less stringent conditions, with primers recognizing a broad range of rabies variants followed by digestion with either restriction enzyme Desulfovibrio desulfuricans I (Dde I) or Haemophilus influenzae Rf . (HinfI), was used to identify the 11 isolates that did not amplify with specific primers (6 DDC, 4 TF and 1 SD) . In addition to these 11 isolates, the less stringent method of amplification, followed by enzyme digestion has identified a total of 125 additional specimens (26 DDC, 94 TF and 5 SD) that were not tested by variant-specific amplification . These data provide a means to track the spread of the different rabies virus variants and allow the ORVP to plan its vaccine disbursement by defining the two epizootic boundaries.

Eur J Clin Microbiol Infect Dis, 1997 Oct, 16(10), 767 - 9
In vitro activity of the new ketolide HMR 3004 compared to an azalide and macrolides against Streptococcus pneumoniae and Haemophilus influenzae; Barry AL et al.; The purpose of this study was to compare the in vitro activity of a new ketolide, HMR 3004 (RU64004), to that of three macrolides and one azalide against 608 Streptococcus pneumoniae and 202 Haemophilus influenzae . Macrolide-resistant pneumococci were susceptible to HMR 3004, even if they were resistant to clindamycin . Against Haemophilus influenzae, HMR 3004 and azithromycin were nearly identical in potency; the macrolides were 8- to 16-fold less active . HMR 3004 may be useful for treating respiratory tract infections if sufficient concentrations can be achieved at the local sites of infection.

Eur J Clin Microbiol Infect Dis, 1997 Oct, 16(10), 750 - 3
Analysis of isolates recovered from multiple sites of the nasopharynx of children colonized by nontypeable Haemophilus influenzae; Bernstein JM et al.; To determine whether the nasopharynx of children is colonized by a single or multiple strains of nontypeable Haemophilus influenzae, cultures were obtained from six nasopharyngeal sites in five children . For each child, all isolates yielded identical polymerase chain reaction fingerprints . The results indicate that these children were colonized in the nasopharynx with a single strain of nontypeable Haemophilus influenzae at one time.

Genomics, 1997 Nov 15, 46(1), 24 - 36
Comparison of DNA sequences with protein sequences; Pearson WR et al.; The FASTA package of sequence comparison programs has been expanded to include FASTX and FASTY, which compare a DNA sequence to a protein sequence database, translating the DNA sequence in three frames and aligning the translated DNA sequence to each sequence in the protein database, allowing gaps and frameshifts . Also new are TFASTX and TFASTY, which compare a protein sequence to a DNA sequence database, translating each sequence in the DNA database in six frames and scoring alignments with gaps and frameshifts . FASTX and TFASTX allow only frameshifts between codons, while FASTY and TFASTY allow substitutions or frameshifts within a codon . We examined the performance of FASTX and FASTY using different gap-opening, gap-extension, frameshift, and nucleotide substitution penalties . In general, FASTX and FASTY perform equivalently when query sequences contain 0-10% errors . We also evaluated the statistical estimates reported by FASTX and FASTY . These estimates are quite accurate, except when an out-of-frame translation produces a low-complexity protein sequence . We used FASTX to scan the Mycoplasma genitalium, Haemophilus influenzae, and Methanococcus jannaschii genomes for unidentified or misidentified protein-coding genes . We found at least 9 new protein-coding genes in the three genomes and at least 35 genes with potentially incorrect boundaries.

J Acquir Immune Defic Syndr Hum Retrovirol, 1997 Dec 1, 16(4), 293 - 300
The association of herpes simplex virus type 2 (HSV-2), Haemophilus ducreyi, and syphilis with HIV infection in young men in northern Thailand; Nelson KE et al.; To evaluate the association between sexually transmitted diseases that commonly may cause genital ulceration and prevalent and incident HIV infections, we conducted three case control studies in a cohort of 21-year-old male military conscripts in northern Thailand . The men were evaluated at baseline in 1991 and semiannually until their discharge 2 years later . Serologic evidence of infection with herpes simplex virus type 2 (HSV-2), Haemophilus ducreyi, and HIV were more frequent at baseline in 83 men with a history of genital ulcer than in 97 men without such a history . Seropositivity to H . ducreyi (odds ratio {OR} = 3.46), HSV-2 (OR = 3.83), and syphilis (OR = 1.53) were more common in HIV-positive than HIV-negative men . Men (N = 45) who seroconverted to HIV while in the military were more often seropositive for H . ducreyi and HSV-2 before HIV seroconversion and also were more likely to seroconvert to HSV-2 and H . ducreyi during the same interval as their HIV seroconversion compared with men who remained HIV-negative . These data suggest that HSV-2 and H . ducreyi may be both markers for high-risk sexual behavior and risk factors for HIV infection among young men in ThailandPIP: Three case-control studies conducted in 1991-93 in a cohort of 21-year-old male military conscripts in northern Thailand investigated the association between HIV infection and three sexually transmitted diseases (STDs) commonly associated with genital ulceration: herpes simplex virus type 2 (HSV-2), Haemophilus ducreyi, and Treponema pallidum . The studies compared 83 men with a history of genital ulcer disease (GUD) at baseline and 97 men without such a history, 103 men who were HIV-positive at baseline and 110 randomly selected HIV-negative conscripts, and 45 men who seroconverted to HIV while they were in the military and 124 men who remained HIV-negative throughout military service . The first study detected a significant dose-response association between number of commercial sex worker visits in the past year, lifetime number of sexual partners, and a history of GUD . Among men with GUD at baseline compared with those without GUD, the odds ratios were 2.52 for HSV-2, 2.02 for H . ducreyi, 0.97 for syphilis, and 2.14 for HIV . In the second study, HIV-infected men were significantly more likely than HIV-negative men to have antibodies to HSV-2 and H . ducreyi and a history of syphilis or gonorrhea . In the third study, men who converted to HIV were significantly more likely to have antibodies to H . ducreyi and HSV-2 at the visit before seroconversion than those who remained HIV-negative . Two independent predictors of seroconversion--HSV-2 seropositivity before conversion and 10 or more lifetime sexual partners--were identified . In northern Thailand, GUD, especially H . ducreyi and HSV-2, may be both a marker for increased HIV risk and a cofactor for HIV transmission .

J Paediatr Child Health, 1997 Oct, 33(5), 422 - 5
Young Malaysian children with lower respiratory tract infections show low incidence of chlamydial infection; Ngeow YF et al.; OBJECTIVE: The incidence of Chlamydia pneumoniae and Chlamydia trachomatis infection was studied among infants and young children admitted to hospital for the management of lower respiratory tract infections, over a 12 month period . METHODOLOGY: Respiratory secretions were examined for chlamydiae by cell culture, enzyme-linked immunosorbent assay and polymerase chain reaction-enzyme immunoassay . Sera were tested by micro-immunofluorescence for chlamydial IgG, IgM and IgA . Other bacterial and viral pathogens were also looked for by standard cultural and serological methods . RESULTS: Of 87 patients aged 2 months-3 years, an aetiologic diagnosis was made in 41 (47.1%) . C . pneumoniae and C . trachomatis were each detected in 1 (1.2%) of the patients . Among common bacterial pathogens, Haemophilus influenzae (13.8%) and Streptococcus pneumoniae (8.1%) were the most frequently identified . Respiratory viruses and elevated Mycoplasma pneumoniae antibodies were found in 10.3% and 9.1% of patients, respectively . CONCLUSION: Chlamydiae are infrequent causes of community-acquired acute lower respiratory tract infections in infants and very young children in Malaysia.

J Paediatr Child Health, 1997 Oct, 33(5), 418 - 21
Incidence of apnoea and bradycardia in preterm infants following DTPw and Hib immunization: a prospective study; Botham SJ et al.; OBJECTIVE: To evaluate the incidence and severity of apnoea and bradycardia in hospitalized preterm infants following immunization at 2 months of age, and identify risk factors . METHODOLOGY: A prospective study of 98 preterm infants, of gestational age 24-31 weeks, immunized at approximately 2 months post natal age with diphtheria-tetanus-whole cell pertussis vaccine (DTPw) in the neonatal intensive care unit (NICU) at King George V Hospital Sydney . Half the infants also received Haemophilus influenzae type b conjugate vaccine (Hib) simultaneously . All infants were monitored for apnoea and bradycardia in the 24 h periods pre- and post immunization . RESULTS: Only one infant had apnoea and/or bradycardia pre-immunization compared with 17 post immunization . For 12 infants these events were brief, self-limiting and not associated with desaturations (oxygen saturation < 90%) . However, for five infants (30%) these events were associated with oxygen desaturation and two of these infants required supplemental oxygen . The group that had apnoea and/or bradycardia and the group that did not were not significantly different in terms of gestational age, birth weight and other variables . Infants who received Hib together with DTPw were less likely to have apnoea and/or bradycardia than those given DTPw alone . CONCLUSION: When considering immunization for preterm infants, the benefits of early immunization must be balanced against the risk of apnoea and bradycardia . We recommend that the cardio-respiratory function of hospitalized infants born at less than 31 weeks gestation be monitored for 48 h post immunization.

J Paediatr Child Health, 1997 Oct, 33(5), 413 - 7
Primary course immunogenicity and reactogenicity of a new diphtheria-tetanus-whole cell pertussis vaccine (DTPw); Nolan T et al.; OBJECTIVE: To establish safety, immunogenicity, and batch stability of a reformulated whole cell pertussis based diphtheriatetanus-whole cell pertussis (DTP) vaccine (nDTPw) compared to the currently marketed Australian DTPw vaccine (Triple Antigen) in a three dose 2, 4 and 6 month primary immunization course . Reformulation was necessary to make the DTPw vaccine suitable for combination with hepatitis B and Haemophilus influenzae b vaccines . METHODS: Double blind randomized controlled trial in suburban Melbourne in 812 healthy infants recruited through maternal and child health centres, of whom 208 received Triple Antigen and 604 received nDTPw . RESULTS: Results for both reactogenicity and immunogenicity were similar and were not significantly different for the three batches of nDTPw . No new, serious, or unexpected adverse effect was recorded . Nearly twice as many nDTPw infants experienced no general reaction to the third dose (18%) compared to Triple Antigen (11%, P = 0.06) . An elevated temperature (> or = 38 degrees C axillary) occurred in about three out of 10 babies overall, with rates being slightly higher for both vaccines after the second vaccination . Local reaction rates were significantly less common for nDTPw on days 2 and 3 following each of the three vaccinations . After dose three, 30% of nDTPw subjects experienced no local reaction compared to 20% of Triple Antigen subjects (P = 0.015, 95% Cl on difference 2%, 19%) . Swelling after doses one and three occurred in 30% and 24% of Triple Antigen subjects, compared to 23% (P = 0.07, 95% Cl diff 0%, 13%) and 14% (P = 0.017, 95% Cl diff 2%, 17%) of nDTPw subjects . Tenderness after doses two and three occurred in 80%, and 78% of Triple Antigen subjects, compared to 71% (P = 0.04, 95% Cl diff 1%, 17%) and 68% (P = 0.025, 95% Cl diff 2%, 19%) of nDTPw subjects . There was a significantly higher post immunization diphtheria antitoxin GMT (2.73 IU/mL) for Triple Antigen compared to nDTPw (1.89 IU/mL; P = 0.02), although no subject in either vaccine group had a tetanus or diphtheria antibody titre less than six times the protective level of 0.01 IU/mL following immunization . The nDTPw post immunization GMTs were significantly higher for Agg2, Fha, and pertactin compared to Triple Antigen geometric mean titres (GMTs) . CONCLUSION: nDTPw is a safe and immunogenic vaccine when compared to Triple Antigen . The reformulated vaccine is an acceptable replacement for the currently marketed formulation, and for evaluation as a component of future combination vaccines.

Clin Immunol Immunopathol, 1997 Dec, 85(3), 236 - 45
Cytokine mRNA profiles during the course of experimental Haemophilus influenzae bacterial meningitis; Diab A et al.; Intraperitoneal inoculation of Haemophilus influenzae type b (Hib) to 3-week-old Sprague-Dawley rats resulted in nonlethal meningitis with high levels of leukocytes in the cerebrospinal fluid (CSF) and positive bacterial culture . Using in situ hybridization, levels of cytokine mRNA-expressing cells were determined in the brain, CSF, and spleen from Hib-inoculated and uninfected control rats . IFN-gamma, IL-1 beta, IL-4, IL-6, IL-10, IL-12, and TNF-alpha mRNA levels were elevated at 12 hr postinoculation (pi) in spleen and CSF . At this time point, strong expression of IL-6 and TGF-beta was detected in the brain, and also of IL-10 at 48 hr while IFN-gamma and IL-12 were expressed at very low levels throughout the observation time . Delayed cytokine induction occurred in CSF compared to spleen and brain . TGF-beta was high in CSF at 48 hr, and some elevation of IL-1 beta, IL-6, IL-10, TNF-alpha, IFN-gamma, and IL-12 was evident at 72 hr pi . This may suggest measures that promote production of TGF-beta and/or IL-10 should be evaluated in treatment of bacterial meningitis.

Changgeng Yi Xue Za Zhi, 1997 Sep, 20(3), 246 - 50
Fatal Haemophilus influenzae pneumonia: two cases report; Wong KS et al.; Haemophilus influenzae is a common cause of acute childhood pneumonia . Most Haemophilus pneumonia generally follow a benign course with occasional complications of pleural effusion, pneumothorax or pneumatocele . Deaths following invasive Haemophilus pneumonia have rarely been reported in children older than 3 years of age . We report 2 deaths in children presenting with fulminant pneumonia, complicated by sepsis and adult respiratory distress syndrome despite vigorous antibiotic therapy and full resuscitative measures.

Jpn J Antibiot, 1997 Sep, 50(9), 768 - 75
{Recent trends in incidence of respiratory tract pathogens and antimicrobial susceptibilities of Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis isolated in 1994 and 1995}; Nishioka K et al.; The incidence of pathogenic bacteria in respiratory tract infections in 1994 and 1995 was investigated using quantitative cultures of sputa from patients with the infections in our department . Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis were isolated at high rates (70.5% in 1994 and 73.8% in 1995) from the specimens of out-patients, and the incident rates were similar to the past data . The antimicrobial susceptibilities of these three pathogens were examined with the agar dilution method . The incidence of penicillin (Pc) resistant S . pneumoniae against which MIC of Pc-G was higher than 0.125 microgram/ml was markedly increased from 24% in 1994 to 34.9% in 1995 . Most of the Pc resistant isolates were also resistant to other antibiotics including erythromycin, minocycline and tosufloxacin . Serotype of strains against which MIC of Pc-G was higher than 1.0 microgram/ml was 19 . The ratios of beta-lactamase-producing strains among H . influenzae isolated in 1994 and 1995 were 20 and 15.8%, respectively, which were slightly higher than those in the past . One quinolone resistant strain was isolated in this study . Although the ratio of beta-lactamase-producing strains among M . catarrhalis was as high (96.7%) as in the past, no increased resistance against the drugs examined was observed.

Jpn J Antibiot, 1997 Sep, 50(9), 756 - 67
{Clinical and bacteriological effects of cefetamet pivoxil against community-acquired respiratory tract infections . Part II}; 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 . That method was approximately equal to that of investigation in 1994 . 1 . Of the 431 respiratory tract infection cases that were treated with CEMT-PI according to a same protocol at a total of 41 institutions in Tokyo, Kanagawa-ken, Saitama-ken and Chiba-ken from January to the beginning of March 1996 . Outpatients accounted for 98.1% of the subjects . Regarding genders to patients, slightly more females (52.6%) than males were included . Diagnoses given to these patients included pharyngo-laryngitis (53.5%), tonsillitis (20.4%) and acute bronchitis (19.1%) . 2 . We investigated clinical efficacy rates (the ratio of those excellent + good) classified by diseases . The improvement rates of pharyngo-laryngitis, tonsillitis and acute bronchitis were more than 85.0% . Other cases were small in number . That of chronic bronchitis-acute increasing change for the worse was 66.7%, pneumonia was 50.0% and bronchiectasis infection was 16.7% . It was not studied that clinical efficacy rates among those who were treated with 1 CEMT-PI tablet twice and among those who were given 2 tablets twice were significant level . 3 . 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 (MICs) and investigation of beta-lactamase production were conducted all together at section of studies, Tokyo Clinical Research Center . Suspected causative bacteria were detected from 274 (63.6%) cases . They included 88 strains of Haemophilus influenzae, 47 strains of Streptococcus pneumoniae, 42 strains of Streptococcus pyogenes, 20 strains of Moraxella subgenus Branhamella catarrhalis and 17 strains of Klebsiella pneumoniae subsp . pneumoniae . Suspected causative bacteria classified by diseases were S . pyogenes (tonsillitis), S . pneumoniae (acute bronchitis and secondary infection of chronic respiratory infection) and H . influenzae (pharyngo-laryngitis), and the detection frequency of those was high . The clinical efficacies (the ratio of improvement) classified by suspected causative bacteria were 84.4% against organism that was indicating CEMT and were 69.2% against organism that was not indicating CEMT.

Infect Immun, 1997 Dec, 65(12), 5017 - 27
Variable number of tandem repeats in clinical strains of Haemophilus influenzae; van Belkum A et al.; An algorithm capable of identifying short repeat motifs was developed and used to screen the whole genome sequence available for Haemophilus influenzae, since some of these repeats have been shown to affect bacterial virulence . Various di- to hexanucleotide repeats were identified, confirming and extending previous findings on the existence of variable-number-of-tandem-repeat loci (VNTRs) . Repeats with units of 7 or 8 nucleotides were not encountered . For all of the 3- to 6-nucleotide repeats in the H . influenzae chromosome, PCR tests capable of detecting allelic polymorphisms were designed . Fourteen of 18 of the potential VNTRs were indeed highly polymorphic when different strains were screened . Two of the potential VNTRs appeared to be short and homogeneous in length; another one may be specific for the H . influenzae Rd strain only . One of the primer sets generated fingerprint-type DNA banding patterns . The various repeat types differed with respect to intrinsic stability as well . It was noted for separate colonies derived from a single clinical specimen or strains passaged for several weeks on chocolate agar plates that the lengths of the VNTRs did not change . When several strains from different patients infected during an outbreak of lung disease were analyzed, increased but limited variation was encountered in all VNTR sites analyzed . One of the 5-nucleotide VNTRs proved to be hypervariable . This variability may reflect the molecular basis of a mechanism used by H . influenzae bacteria to successfully colonize and infect different human individuals.

Infect Immun, 1997 Dec, 65(12), 5010 - 6
The acylated form of protein D of Haemophilus influenzae is more immunogenic than the nonacylated form and elicits an adjuvant effect when it is used as a carrier conjugated to polyribosyl ribitol phosphate; Akkoyunlu M et al.; The nonacylated form of protein D (PDm) of Haemophilus influenzae has been shown to induce the production of antibodies that are bactericidal to homologous and heterologous nontypeable H . influenzae (NTHi) strains . In this study, immunization of rats with lipoprotein D (LPD) induced higher levels of anti-protein D immunoglobulin G and A serum antibodies than immunization with PDm, and the bactericidal activities of sera from LPD-immunized rats were greater than those of sera from PDm-immunized rats . Immunization with LPD or PDm did not prevent the development of acute otitis media (AOM) when rats were challenged with 10(4) CFU of an NTHi strain . However, on the eighth day of bacterial challenge, 50% (5 of 10) of LPD-immunized rats had recovered from otitis media and 30% (3 of 10) had negative middle ear cultures, whereas only 30% (3 of 10) of PDm-immunized rats had recovered, though none was culture positive . Immunization with an inactivated homologous bacterial strain elicited 70% protection (i.e., 7 of 10 rats) in the rat otitis media model . LPD and PDm were also conjugated to the H . influenzae type b (Hib) capsular polysaccharide, polyribosyl ribitol phosphate (PRP), to test protein D-conjugated PRP vaccine's potential for protection against Hib infection . When two LPD-conjugated and two PDm-conjugated PRP vaccines, each containing a different protein concentration, and a tetanus toxoid-conjugated vaccine (ACT-HIB) were tested in the experimental model of rat otitis induced with a Hib strain (Minn A), both of the LPD-conjugated and one of the PDm-conjugated vaccines induced significant protection from AOM, the level of protection being highest in animals given the vaccine with the highest LPD content . Sera from these rats also manifested the highest anti-PRP and anti-LPD antibody levels and the highest bactericidal activities against a Hib strain and an NTHi strain.

Infect Immun, 1997 Dec, 65(12), 4918 - 25
A strategy for rational design of fully synthetic glycopeptide conjugate vaccines; Chong P et al.; The present study describes a strategy to rationally design fully synthetic glycopeptide conjugate vaccines . Glycopeptide immunogens were constructed by coupling synthetic oligosaccharides comprising repeating units of synthetic 3-beta-D-ribose-(1-1)-D-ribitol-5-phosphate (sPRP) to synthetic peptides containing potent T-helper cell determinants and B-cell epitopes of the Haemophilus influenzae type b (Hib) outer membrane proteins (OMPs) P1, P2, and P6 . Rabbit immunogenicity studies revealed that some of these fully synthetic glycoconjugates were capable of eliciting high titers of both anti-PRP and anti-OMP immunoglobulin G antibodies . In addition, we systematically investigated the factors which could influence their immunogenicity . We observed that the magnitude of the anti-PRP antibody response markedly depended on the relative spatial orientation of sPRP and T-cell epitopes, the anti-PRP antibody response was enhanced when a multiple antigenic peptide was used as a carrier, the anti-PRP antibody response was optimal for three PRP repeating units, and lipidation of peptide-PRP conjugates had a minimal effect on the magnitude of the anti-PRP antibody response . The results of this study clearly demonstrate that coupling a carbohydrate hapten to a peptide can provide T-cell help and convert it into a T-cell-dependent antigen . The antisera raised against these conjugates were also found to be protective against Hib infection in the infant rat model of bacteremia.

J Bacteriol, 1997 Dec, 179(23), 7315 - 20
In vitro Tn7 mutagenesis of Haemophilus influenzae Rd and characterization of the role of atpA in transformation; Gwinn ML et al.; Haemophilus influenzae Rd is a gram-negative bacterium capable of natural DNA transformation . The competent state occurs naturally in late exponential growth or can be induced by a nutritional downshift or by transient anaerobiosis . The genes cya, crp, topA, and sxy (tfoX) are known to function in the regulation of competence development . The phosphoenolpyruvate:carbohydrate phosphotransferase system functions to maintain levels of cyclic AMP necessary for competence development but is not directly involved in regulation . The exact signal(s) for competence and the genes that mediate the signal(s) are still unknown . In an effort to find additional regulatory genes, H . influenzae Rd was mutated by using an in vitro Tn7 system and screened for mutants with a reduced ability to induce the competence-regulatory gene, comA . Insertions in atpA, a gene coding for the alpha subunit of the F1 cytoplasmic domain of the ATP synthase, reduce transformation frequencies about 20-fold and cause a significant reduction in expression of competence-regulatory genes, while the expression of constitutive competence genes is only minimally affected . In addition, we found that an insertion in atpB, which encodes the a subunit of the F0 membrane-spanning domain, has a similar effect on transformation frequencies.

J Clin Pathol, 1997 Sep, 50(9), 765 - 8
Haemophilus influenzae: an underrated cause of vulvovaginitis in young girls; Cox RA; AIMS: To establish the common pathogens associated with infective vulvovaginitis in young girls in the local population and to determine current management of this condition . METHODS: A prospective laboratory based survey was carried out over 19 months . A questionnaire was then sent to local general practitioners and hospital doctors . RESULTS: One hundred and six swabs were received during the study period of which 43 (40.5%) yielded organisms recognised as causes of vulvovaginitis . The most common pathogen was group A beta haemolytic streptococcus (19), with Haemophilus influenzae the second most common (11) . Candida was isolated on nine occasions . The users' questionnaire had an overall response rate of 52% . Forty one per cent of respondents nominated candida as the most common cause of this condition . Forty six per cent were aware that beta haemolytic streptococci caused juvenile vulvovaginitis, but only four (3.6%) knew that H influenzae was a possible pathogen . The most popular agent for empirical treatment of vulvovaginitis was topical clotrimazole cream, although 24 respondents (22%) prescribed antibiotics that are active against both group A beta haemolytic streptococci and H influenzae . CONCLUSIONS: Although H influenzae is the second most common infective cause of juvenile vulvovaginitis in the local population, most doctors managing these patients were unaware of its importance and may not be prescribing appropriate empirical treatment.

Eur Respir J, 1997 Oct, 10(10), 2327 - 33
Iron-binding proteins in sputum of chronic bronchitis patients with Haemophilus influenzae infections; Vogel L et al.; Airway inflammation during infection is associated with increased transudation of serum proteins and increased production of protein by the airway epithelium . We therefore, assessed whether Haemophilus influenzae infections in patients with chronic bronchitis are associated with increased levels of transferrin and lactoferrin in the sputum compared to uninfected patients . Sputum sol phase and serum samples from 14 infected and 13 uninfected patients with chronic bronchitis and from 12 bronchial asthma patients were included in the study . The median Q-values (the concentration in sputum sol phase/the concentration in serum) x 10(3) of transferrin appeared increased in chronic bronchitis patients with an H . influenzae infection (26.0, n=13) compared to uninfected controls (9.5, n=11) and bronchial asthma patients (4.5, n=6) . The ratio of the Q(transferrin)/Q(albumin) was >1 in infected chronic bronchitis patients, indicating local production of transferrin . Growth of H . influenzae was stimulated more in sputum from infected and uninfected patients with chronic bronchitis than in sputum from patients with bronchial asthma . The concentrations of lactoferrin were not significantly different in infected (n=14) and uninfected (n=13) chronic bronchitis patients and bronchial asthma patients (n=12) (median 137.4, 84.6, 87.1 mg x L(-1), respectively) . We conclude that in patients with chronic bronchitis with Haemophilus influenzae infections, the levels of transferrin are increased and the levels of lactoferrin are not associated with infections.

Eur Respir J, 1997 Oct, 10(10), 2319 - 26
Persisting Haemophilus influenzae strains induce lower levels of interleukin-6 and interleukin-8 in H292 lung epithelial cells than nonpersisting strains; Bresser P et al.; Nonencapsulated Haemophilus influenzae strains isolated from patients with chronic bronchitis can be divided into those that persist in the lower respiratory tract and those that do not . We tested the hypothesis that persisting and nonpersisting strains differ in the extent to which they activate epithelial cells to produce two potent inflammatory mediators, interleukin (IL)-6 and IL-8 . A suspension of 10(7) and 10(8) colony forming units (cfu) x mL(-1) of H . influenzae, persisting and nonpersisting, induced a dose- and time-dependent production of IL-6 and IL-8 by the human pulmonary mucoepidermoid carcinoma-derived cell line H292, but levels of IL-6 were lower after exposure to persisting H . influenzae (p<0.05) . IL-8 production showed a similar trend (p<0.02; analysis of variance) . H . influenzae bacteria that adhered to H292 cells were equally distributed over persisting and nonpersisting isolates and induced IL-6 and IL-8 levels similar to their nonadhering counterparts . The difference between persisting and nonpersisting H . influenzae was not due to cytotoxic, antimetabolic or antiproliferative effects on H292 cells . Furthermore, pre-exposure of cells to persisting and nonpersisting isolates did not block subsequent IL-1beta-induced IL-6 production . We conclude that persisting clinical isolates induce less interleukin-6 and interleukin-8 in H292 cells than nonpersisting isolates, probably because they excrete lower amounts of a stimulus of H292 cells . The stimulus is heat stable, hydrophilic and nonproteinous and probably not lipopolysaccharide alone . These findings support the suggestion that some strains of Haemophilus influenzae that persist in the airways of patients, may do so because they induce only a weak inflammatory response.

Antimicrob Agents Chemother, 1997 Nov, 41(11), 2582 - 5
In vitro and in vivo antibacterial activities of CS-940, a new fluoroquinolone, against isolates from patients with respiratory infections; Miyazaki S et al.; We compared the in vivo and in vitro activities of CS-940, a new fluoroquinolone, with those of a group of other drugs . The activities of CS-940 against gram-positive cocci and gram-negative rods, including methicillin-susceptible Staphylococcus aureus and penicillin-resistant Streptococcus pneumoniae, were comparable to those of tosufloxacin, with MICs at which 90% of the strains were inhibited (MIC90s) of 0.5 microg/ml or less . Against methicillin-resistant S . aureus, CS-940 was as active as tosufloxacin, with a MIC90 of 16 microg/ml . The efficacy of CS-940 against murine respiratory infections due to S . pneumoniae or Haemophilus influenzae was better than those of tosufloxacin and sparfloxacin . The efficacy of oral doses of CS-940 reflected not only potent in vitro activity but also a high transmigration ratio from the bloodstream to lung tissues.

Antimicrob Agents Chemother, 1997 Nov, 41(11), 2522 - 6
Studies of the killing kinetics of benzylpenicillin, cefuroxime, azithromycin, and sparfloxacin on bacteria in the postantibiotic phase; Odenholt I et al.; Most antibiotics are known to be incapable of killing nongrowing or slowly growing bacteria with few exceptions . Bacterial cell division is inhibited during the postantibiotic phase (PA phase) after short exposure to antibiotics . Only scarce and conflicting data are available concerning the ability of antibiotics to kill bacteria in the PA phase . The aim of the present study was to investigate the killing effect of four different antibiotics on bacteria in the PA phase . A postantibiotic effect (PAE) was induced by exposing Streptococcus pyogenes and Haemophilus influenzae to 10x MICs of benzylpenicillin, cefuroxime, sparfloxacin, and azithromycin . The bacteria were thereafter reexposed to a 10x MIC of the same antibiotic used for the induction of the PAE at the beginning of and after 2 and 4 h in the PA phase . Due to a very long PAE, the bacteria in PA phase induced by azithromycin were also exposed to 10x MICs after 6 and 8 h . A previously unexposed culture exposed to a 10x MIC was used as a control . The results seem to be dependent on both the antibiotic used and the bacterial species . The antibiotics exhibiting a fork bactericidal action gave significantly reduced killing of the bacteria in PA phase (cefuroxime with S . pyogenes, P < 0.01, and sparfloxacin with H . influenzae, P < 0.001), which was restored at 4 h for cefuroxime with S . pyogenes . There was a tendency to restoration of the bactericidal activity also with sparfloxacin and H . influenzae, but there was still a significant difference in killing between the control and the test bacteria in PA phase at 4 h . However, in the combinations with a lesser bactericidal effect (benzylpenicillin with S . pyogenes and sparfloxacin with S . pyogenes), there was no difference in killing between the control and the test bacteria in PA phase . Azithromycin induced long PAEs in both S . pyogenes and H . influenzae and exhibited a slower bactericidal action on both the control and the bacteria in PA phase especially at the end of the PAE, when the killing was almost bacteriostatic . Our findings in this study support the concept that a long interval (> 12 h) between doses of azithromycin, restoring full bactericidal action, may be beneficial to optimize efficacy of this drug but is not necessary for the other antibiotics evaluated, since the bactericidal effect seems to be restored already at 4 h.

Biochim Biophys Acta, 1997 Sep 4, 1348(1-2), 228 - 35
Phosphatidylserine synthase from yeast; Yamashita S et al.; Whereas mammalian cells produce PS by a base exchange reaction from preexisting phospholipids, yeast cells synthesize PS from CDP-diacylglycerol and serine by the PS synthase reaction . Yeast PS synthase was purified to homogeneity and shown to have a molecular mass of 23 kDa . The activity is dependent on either Mg2+ or Mn2+ and Triton X-100 . The enzyme specifically transfers the phosphatidyl group from CDP-diacylglycerol or dCDP-diacylglycerol to L-serine, but not to threonine, cysteine and ethanolamine . The PSS/CHO1 gene encoding the enzyme was cloned by the complementation of the choline auxotrophic pss/cho1 mutant . The deduced protein comprises 279 amino acids with a calculated molecular mass of 30,804 . The primary translate undergoes proteolytic processing to the enzymatically more active 23-kDa enzyme . The deduced amino acid sequence contains several putative membrane-spanning regions and resembles that of the Bacillus subtilis enzyme, but not those of the E . coli and Haemophilus influenzae enzymes . The sequence also contains the local, conserved region found in enzymes catalyzing the transfer of the phosphoalcohol moiety from CDP-alcohol, such as PI synthase, cholinephosphotransferase and phosphatidylglycerolphosphate synthase . The activity of PS synthase is maximal in the exponential phase, but decreases when cells enter the stationary phase . The enzyme is phosphorylated at a single serine residue by cyclic AMP-dependent protein kinase with a 60-70% decrease in enzymatic activity, but the primary translation product is not phosphorylated . PS synthase is inhibited by CTP, probably due to the chelation of the divalent cations, Mg2+ and Mn2+, and also by sphingoid bases, such as sphinganine and phytosphingosine . Phosphatidate, phosphatidylcholine and phosphatidylinositol are stimulatory, whereas cardiolipin and diacylglycerol are inhibitory . The expression of yeast PS synthase is transcriptionally repressed by myo-inositol and choline in a coordinate manner with other phospholipid-synthesizing enzymes . The upstream regulatory region of the PSS/CHO1 gene responsible for the myo-inositol-choline regulation was identified . An octameric sequence, CATRTGAA (R = A or G), plays an important role in the conferral of the myo-inositol-choline transcriptional regulation.

FEMS Microbiol Lett, 1997 Nov 1, 156(1), 165 - 70
Human lactoferrin receptor activity in non-encapsulated Haemophilus influenzae; Vogel L et al.; Since the ability of bacteria to compete with lactoferrin for iron contributes to the pathogenesis of mucosal infections, the presence of lactoferrin receptor activity in non-encapsulated Haemophilus influenzae was investigated . The growth of 18 H . influenzae isolates from the sputum samples of chronic bronchitis patients and of six of seven H . influenzae throat isolates from healthy adults was stimulated by iron saturated human lactoferrin . Apo-lactoferrin did not stimulate the growth of H . influenzae . Human lactoferrin binding to iron limited bacteria was detected for 16 H . influenzae strains from chronic bronchitis patients and for five of seven isolates from healthy adults . We conclude that the majority of H . influenzae isolates tested bind human lactoferrin and that the iron from lactoferrin is used for growth.

Biochem Biophys Res Commun, 1997 Oct 29, 239(3), 835 - 9
Isolation of ALU1-P gene encoding a protein with aluminum tolerance activity from Arthrobacter viscosus; Jo J et al.; We isolated a DNA fragment (ALU1-P) encoding a protein with an activity of aluminum tolerance from an Al tolerant soil microorganism, Arthrobacter viscosus . This microorganism was isolated from acidic tea field soils . The cloned DNA is composed of 1090 nucleotides, which has one open-reading frame without any stop codon . However, when the DNA fragment was transferred into Escherichia coli, a microorganism susceptible to Al toxicity, it endowed E . coli with Al tolerance . The deduced amino acid sequence of the DNA showed 65% identity with the protein of YbaX gene in Escherichia coli, and 51.1% identity with YB91 Haein hypothetical protein of HI1191 gene in Haemophilus influenzae . The ALU1-P gene in the expression vector produced a protein of 192 amino acids deriving a molecular weight of 21.3 kDa by using the stop codon in vector . The ALU1-P gene is a new one that has the characteristic of Al tolerance.

Mol Microbiol, 1997 Sep, 25(5), 979 - 87
Characterization of a ferric-binding protein mutant in Haemophilus influenzae; Kirby SD et al.; Ferric-binding proteins (FbpA) have been implicated in the transferrin receptor-mediated iron acquisition pathways of Haemophilus influenzae and Neisseria spp . These proteins are believed to function by shuttling iron from outer membrane transferrin receptors to a specific inner membrane permease complex . However, the role of these proteins has not been conclusively resolved, as attempts at creating isogenic mutants in the fbpA genes of both species have been unsuccessful, prompting the hypothesis that FbpA may play a critical role in H . influenzae and Neisseria spp . This study describes the construction and characterization of an H . influenzae isogenic fbpA mutant . It is demonstrated that this mutant is deficient in its ability to use human transferrin as a sole iron source, even though the strain is still competent for binding human transferrin . It is also demonstrated that this mutant is impaired in its ability to use ferric citrate as an iron source, and grows at a reduced rate relative to wild type in broth supplemented with protoporphyrin rather than haemin.

J Med Microbiol, 1997 Oct, 46(10), 883 - 5
Cefsulodin chocolate blood agar: a selective medium for the recovery of Haemophilus influenzae from the respiratory secretions of patients with cystic fibrosis; Smith A et al.; A modified chocolate blood agar medium incorporating cefsulodin, a semi-synthetic cephalosporin, was developed and compared with non-selective chocolate blood agar and selective haemin-bacitracin blood agar for the routine isolation of Haemophilus influenzae from the respiratory secretions of patients with cystic fibrosis . The results showed that cefsulodin chocolate blood agar improved the recovery rate of H . influenzae in this group of patients . The medium was stable on storage for 10 days at 4 degrees C.

Acta Paediatr Jpn, 1997 Oct, 39(5), 541 - 5
Nasopharyngeal colonization with Haemophilus influenzae type b among infants and children in Japan; Kuroki H et al.; Healthy carriers of Haemophilus influenzae type b (Hib) play an important role in the spread of invasive Hib disease . The aim of the present study was to estimate Hib colonization among infants and children in Japan . Specimens from throat and nasopharyngeal cultures were obtained by thorough swabbing of both tonsils and the posterior pharynx . Specimens were inoculated on Hib antiserum agar . This was prepared with Levinthal base and Hib antiserum . Conventional methods were used concomitantly . Four of 474 infants from 1-48 months of age (0.84%) had Hib cultured from their nasopharynx . The carriage rate in 1-12 months old infants was 0.62% (2/322 cases), and that in 13-48 month old children was 1.32% (2/152 cases) . Five of 167 (3.0%) 13-year-old children, and five of 154 (3.2%) 9-year-old children were asymptomatic carriers . Thirty-five of 104 household contacts of a patient with invasive Hib disease (33.6%) had Hib colonization . The carriage rate in healthy Japanese children may not be different from that in the USA prior to the availability of the conjugate Hib vaccine . The Hib carriage rate in household contacts of patients with invasive Hib disease was higher than in healthy children (P < 0.005) . Our results suggest the possibility of an outbreak of invasive Hib disease in Japan.

Curr Opin Pulm Med, 1996 May, 2(3), 228 - 35
Prevention of community-acquired and nosocomial pneumonia; Simberkoff MS et al.; Pneumonia is an important cause of morbidity and mortality in the United States . The provision of effective prophylaxis for pneumonia has become a major goal for both public health officials and individual physicians . Prophylaxis for community-acquired pneumonia is pathogen-specific and is directed toward the most common microorganisms that cause it . The 23-valent pneumococcal polysaccharide vaccine; the trivalent influenza vaccine; the Haemophilus b conjugate vaccine; and either trimethoprim-sulfamethoxazole, dapsone, or aerosolized pentamidine are recommended to prevent Streptococcus pneumoniae, influenza viruses, H . influenzae type b, and Pneumocystis carinii respectively . Except for the microorganisms listed above, the prevention of nosocomial pneumonia is not pathogen-specific . Rather, prevention of nosocomial pneumonia requires the use of infection control procedures, including patient and staff education; isolation of patients with highly contagious respiratory pathogens; vigorous hand washing; cleaning and sterilizaton of respiratory equipment; and use of sterile water in nebulizers and humidifiers . It also requires procedures to limit pooling and aspiration of secretions, such as positioning and rotation of the bed-bound patient; frequent suctioning of respiratory secretions using gloves and sterile suction catheters; and limiting enteral alimentation . Finally, selective decontamination of the digestive tract may be considered for intubated patients.

Curr Opin Pulm Med, 1996 May, 2(3), 181 - 5
Infective pathogenesis and outcomes in chronic bronchitis; Ball P; Bacterial infection is an important cause of exacerbations of chronic bronchitis, which can precipitate both direct toxic and host-mediated inflammatory bronchial epithelial damage . Repeated episodes, especially when caused by Haemophilus influenzae (the major pathogen) may be associated with more rapid deterioration in respiratory function . Criteria to identify patients with more severe disease and those who require either hospitalization or intensive care have been developed and can improve mortality . Antibiotic therapy has an accepted place in management, and predictors of the outcome of exacerbations have now been developed that allow severity staging of individual episodes . Nevertheless, despite the recognition of excessive failure rates due to increasing bacterial resistance and poor respiratory kinetics, many patients continue to receive empiric therapy with amoxicillin and other oral beta-lactams . Less empiric management guidelines, produced in Europe and North America and based on such criteria, now provide a framework for rational prescribing . Perhaps more importantly, it is now possible to design and perform placebo- and comparator-controlled studies of new antibacterials, such as the fluoroquinolones and azalides, with improved potency and kinetic properties . This will enable both an assessment of their place in the management of exacerbations in at-risk patients with severe disease and an evaluation of their longer-term role in the prevention of a decline in respiratory function consequent upon repeated and ineffectively treated exacerbations.

Epidemiol Infect, 1997 Oct, 119(2), 167 - 74
Antibody levels against Streptococcus pneumoniae and Haemophilus influenzae type b in a population of splenectomized individuals with varying vaccination status; Konradsen HB et al.; In order to determine antibody levels against Streptococcus pneumoniae (pneumococcus) and Haemophilus influenzae type b (Hib) in a population of splenectomized subjects, 561 persons in a Danish county, splenectomized between 1984 and 1993 were identified . Two hundred and thirty-five were alive and 149 participated in the study . Each person donated a blood sample for antibody determination by ELISA . Though vaccine coverage among the 149 persons was 91% only 52% had 'protective' levels of pneumococcal antibodies . Despite recommendations for regular follow-up on pneumococcal antibody levels this had only been carried out in 4% of the subjects . Splenectomized subjects who needed pneumococcal revaccination were significantly more likely to have received their initial vaccination less than 14 days before or after splenectomy, as recommended, than those not requiring revaccination . Therefore, the timing of initial pneumococcal vaccination in relation to splenectomy seems to be important . All persons had Hib antibody levels higher than 0.15 microgram/ml and 60% had levels higher than 1 microgram/ml, which are the levels thought to provide short term and long term protection, respectively . In total, 37% of the 149 persons tested had pneumococcal and Hib antibody levels thought to correlate with protection from serious infections.

Nat Struct Biol, 1997 Nov, 4(11), 919 - 24
Structure of Haemophilus influenzae Fe(+3)-binding protein reveals convergent evolution within a superfamily; Bruns CM et al.; The first crystal structure of the iron-transporter ferric ion-binding protein from Haemophilus influenzae (hFBP), at 1.6 A resolution, reveals the structural basis for iron uptake and transport required by several important bacterial pathogens . Paradoxically, although hFBP belongs to a protein superfamily which includes human transferrin, iron binding in hFBP and transferrin appears to have developed independently by convergent evolution . Structural comparison of hFBP with other prokaryotic periplasmic transport proteins and the eukaryotic transferrins suggests that these proteins are related by divergent evolution from an anion-binding common ancestor, not from an iron-binding ancestor . The iron binding site of hFBP incorporates a water and an exogenous phosphate ion as iron ligands and exhibits nearly ideal octahedral metal coordination . FBP is highly conserved, required for virulence, and is a nodal point for free iron uptake in several Gram-negative pathogenic bacteria, thus providing a potential target for broad-spectrum antibacterial drug design against human pathogens such as H . influenzae, Neisseria gonorrhoeae, and Neisseria meningitidis.

Scand J Infect Dis, 1997, 29(4), 387 - 92
Recurrence of pneumonia in middle-aged and elderly adults after hospital-treated pneumonia: aetiology and predisposing conditions; Hedlund J et al.; In order to investigate the predisposing conditions and aetiologic agents in patients with recurrent pneumonia, we prospectively studied 653 immunocompetent patients, 50-85 years of age, who had been treated in hospital for community-acquired pneumonia . After an average patient follow-up period of 32 months, 11 variables were examined for association with the following end points: death, recurrence of pneumonia and recurrence of pneumococcal pneumonia . During the follow-up period there were 171 episodes of pneumonia in 115 of the 653 patients, and 52 deaths (all causes) . Multivariate analysis showed that age, male sex, congestive heart failure and presence of other chronic diseases were significantly associated with higher mortality . Age and chronic pulmonary disease were associated with recurrence of pneumonia . The major aetiologic agents were Streptococcus pneumoniae (26%), Haemophilus influenzae (11%) and Moraxella catarrhalis (6%) . We conclude that pneumonia recurrences are common in middle-aged and elderly patients after treatment in hospital for community-acquired pneumonia . The recurrence risk is higher in elderly patients, and in those with chronic pulmonary diseases . Given the prominence of H . influenzae and M . catarrhalis found in the present study, these organisms should always be considered when choosing the initial antibiotic in patients with recurrent pneumonia.

J Infect Dis, 1997 Nov, 176(5), 1253 - 9
Bacteriologic response to oral cephalosporins: are established susceptibility breakpoints appropriate in the case of acute otitis media?
Dagan R, Abramson O, Leibovitz E, Greenberg D, Lang R, Goshen S, Yagupsky P, Leiberman A, Fliss DM.
Bacteriologic response to cefuroxime axetil and cefaclor administered for 10 days was evaluated in acute otitis media (AOM) in patients aged 6-36 months . Middle ear fluid culture was obtained by tympanocentesis before treatment, on day 4 or 5 after initiation of treatment, and if clinical relapse occurred before day 17 . Bacteriologic failure was observed in 32% of patients receiving cefaclor versus 15% of patients receiving cefuroxime axetil (P = .009) . Failure rates increased with increasing MIC: For Streptococcus pneumoniae, 0.5 microg/mL (established as cutoff value for cefuroxime by the National Committee for Clinical Laboratory Standards {NCCLS}) discriminated between success and failure . For Haemophilus influenzae, high failure rates were observed for cefaclor, even with low MICs (< or = 1.0 microg/mL), and with both drugs they tended to increase with increasing MIC, even for values below the cutoff suggested by the NCCLS (8.0 and 4.0 microg/mL for cefaclor and cefuroxime, respectively) . Thus, for AOM caused by H . influenzae, lower susceptibility cutoff levels for MICs should be established.

J Infect Dis, 1997 Nov, 176(5), 1247 - 52
Human immune response to nontypeable Haemophilus influenzae in chronic bronchitis; Yi K et al.; Nontypeable Haemophilus influenzae (NTHI) causes recurrent respiratory tract infections in patients with chronic bronchitis . To elucidate the human immune response to NTHI, sera from 2 patients with exacerbations of chronic bronchitis due to NTHI were characterized . Both patients developed new bactericidal antibodies following infection . Immunoblot assays with homologous strains revealed antibodies to many antigens, with minimal difference between pre- and postexacerbation sera . By contrast, whole cell radioimmunoprecipitation, which detects antibodies exclusively to epitopes exposed on the bacterial surface, revealed that both patients made new antibodies to a limited number of antigens following infection, including P2, the major outer membrane protein of NTHI . Adsorption experiments showed that strain-specific, surface-exposed epitopes on the P2 molecule are targets for bactericidal antibodies . These results indicate that new bactericidal antibodies following infection by NTHI recognize antigenically heterogeneous surface-exposed epitopes on P2 and other surface proteins of NTHI.

J Infect Dis, 1997 Nov, 176(5), 1239 - 46
The effect of interleukin-10 on meningeal inflammation in experimental bacterial meningitis; Paris MM et al.; Interleukin-10 (IL-10) is a cytokine with antiinflammatory effects . In a rabbit model of meningitis, IL-10 was given intracisternally or intravenously to evaluate the impact on inflammation induced by lipooligosaccharide (LOS), Haemophilus influenzae type b (Hib), or Listeria monocytogenes . Intracisternal IL-10 in concentrations >1 microg significantly reduced tumor necrosis factor-alpha (TNF-alpha) and lactate values in cerebrospinal fluid (CSF) . Intravenous IL-10 (1 mg/kg) in two doses after intracisternal LOS significantly reduced CSF TNF-alpha and lactate . When Hib was used, animals were treated with ceftriaxone and dexamethasone with or without IL-10 (1 mg/kg) . TNF-alpha was significantly reduced in animals treated with IL-10, dexamethasone, or both compared with levels in rabbits receiving ceftriaxone alone . Comparable results were obtained when L . monocytogenes was inoculated and animals were treated with ampicillin with or without IL-10, dexamethasone, or nothing . In conclusion, IL-10 modulates CSF TNF-alpha concentrations in experimental LOS, Hib, or L . monocytogenes meningitis . The maximal inhibitory effect was seen when IL-10 and dexamethasone were combined.

Gene, 1997 Oct 15, 199(1-2), 49 - 56
Sequence of the ponA gene and characterization of the penicillin-binding protein 1A of Pseudomonas aeruginosa PAO1; Handfield J et al.; The nucleotide sequence of the ponA gene encoding the high molecular-mass penicillin-binding protein 1A (PBP1A) of Pseudomonas aeruginosa (Pa) PAO1 was determined and characterized . The predicted PBP1A protein of 822 amino acids (aa) has a calculated molecular mass of 91.2 kDa corresponding to the size of the protein expressed in vitro and in vivo . A penicillin-binding (PB) assay showed that the Pa ponA gene product covalently binds penicillin . The deduced PBP1A aa sequence has features typical of class-A high-molecular-mass PBPs: a highly hydrophobic N-terminus portion containing a potential transmembrane segment which might anchor the protein to the cytoplasmic membrane; an N-terminal module with the conserved boxes 1 (E86D(DN)F(AN)H(Y)G), 2 (G117GS(T)I(TM)Q), 3 (R139K(IN)E(ILL)AL) and 4 (R221R(NW)IL); a PB module with the conserved boxes 5 (S461SFK), (S520RN) and (K695TG); an internal extension at aa 297-407 between the N-terminal and PB modules; and a C-extension at the end of the PB module at aa 742 to 822 . The highest percentage of similarity (62.8%) was found with the class A high-molecular-mass PBP1A of Escherichia coli (Ec) and Haemophilus influenzae . The observed extensive homology in the modular design of the Pa PBP1A with the bifunctional Ec PBP1A suggests structural and functional relationships between these proteins and refutes the proposed correspondence between Pa PBP1A and Ec PBP1B.

Clin Infect Dis, 1997 Oct, 25(4), 915 - 7
Invasive Haemophilus influenzae type b (Hib) infection in an adult patient with a selective deficiency of antibody to the Hib capsular polysaccharide; Follin P et al.; A 31-year-old woman without any underlying disease contracted severe invasive Haemophilus influenzae type b (Hib) infection but developed no antibodies to the Hib capsular polysaccharide . Serum immunoglobulin levels were normal, but she had an isolated deficiency of antibody to Hib . Subsequently, immunization with a tetanus toxoid-conjugated Hib vaccine induced only a minimal response . However, she had a protective level of antibody (> 1 microgram/mL) after the fifth vaccination.

Microbiologia, 1997 Sep, 13(3), 309 - 14
Molecular basis of antimicrobial resistance in non-typable Haemophilus influenzae; Sanchez L et al.; Strains of the facultative anaerobe Haemophilus influenzae, both type b and non typable strains, are frequently multiresistant . The measurement of the antibiotic permeability of Haemophilus influenzae outer membrane (OM) shows that antibiotics can cross through the OM easily . Thus, enzymatic activity or efflux pumps could be responsible for multiresistance . An efflux system closely related to AcrAB of Escherichia coli is present in Haemophilus influenzae . However, their role in multiresistance seems irrelevant . Classical mechanisms such as plasmid exchange seems to be playing a major role in the multidrug resistance in Haemophilus influenzae.

Infect Immun, 1997 Nov, 65(11), 4696 - 700
Transcription of genes encoding iron and heme acquisition proteins of Haemophilus influenzae during acute otitis media; Whitby PW et al.; Unencapsulated Haemophilus influenzae is the second most common etiologic agent of otitis media in children . H . influenzae requires heme for aerobic growth in vitro and is able to utilize hemoglobin and complexes of heme-hemopexin, heme-albumin, and hemoglobin-haptoglobin and ferritransferrin as sources of iron and heme in vitro . Several of the acquisition mechanisms have been characterized and been shown to be heme repressible in vitro . However, little is known about the expression of heme and/or iron acquisition mechanisms during infections in the middle ear . This study was performed to determine if the genes encoding heme and iron acquisition proteins are transcribed during in vivo growth and to compare these findings with those for samples grown in vitro . Reverse transcriptase PCR (RT-PCR) was used to analyze total RNA fractions derived from in vitro- and in vivo-grown H . influenzae . Genes encoding the transferrin-binding proteins TbpA and TbpB, the 100-kDa hemopexin-binding protein HxuA, and the hemoglobin-binding protein HgpA were transcribed during otitis media . Twelve middle ear fluid samples were analyzed by blind RT-PCR to determine the transcriptional status of these genes in H . influenzae during otitis media . Five isolates had transcripts corresponding to tbpA, tbpB, and hxuA . The presence of hgpA transcripts was variable, depending on the presence of hgpA in the genome of the H . influenzae isolate . Samples without H . influenzae gene transcripts contained other etiologic agents commonly causing otitis media . These data demonstrate that H . influenzae iron and/or heme acquisition genes are transcribed during otitis media and suggest that the microenvironment during acute otitis media starves H . influenzae of heme.

Infect Immun, 1997 Nov, 65(11), 4675 - 81
Phase variation and conservation of lipooligosaccharide epitopes in Haemophilus somnus; Inzana TJ et al.; The bovine-specific pathogen Haemophilus somnus is capable of undergoing structural and antigenic phase variation in its lipooligosaccharide (LOS) components after in vivo and in vitro passage . However, commensal isolates from the reproductive tract have not been observed to vary in phase (T . J . Inzana, R . P . Gogolewski, and L . B . Corbeil, Infect . Immun . 60:2943-2951, 1992) . We now report that specific monoclonal antibodies (MAbs) to the LOSs of Haemophilus aegyptius, Neisseria gonorrhoeae, and Haemophilus influenzae, as well as H . somnus, reacted with some phase-variable epitopes in H . somnus LOS . All reactive MAbs bound to LOS components of about 4.3 kDa in the same H . somnus isolates, including a non-phase-varying strain . Following in vitro passage of a clonal variant of strain 738 that was nonreactive with the MAbs, 11.8% of young colonies shifted to a reactive phenotype . A digoxigenin-labelled 5'-CAATCAATCAATCAATCAATCAATCAAT-3' oligonucleotide probe hybridized to genomic DNA from strain 738 but did not react with DNA from a non-phase-varying strain . Sequence analysis of the gene containing 5'-CAAT-3' tandem sequences revealed 48% amino acid homology with the lex-2B gene-encoded protein of H . influenzae type b . Our results indicate that some LOS epitopes are conserved between H . somnus and other Haemophilus and Neisseria species, that LOS phase variation may occur at a high rate in some strains of H . somnus, and that phase variation may, in part, be due to 5'-CAAT-3' tandem sequences present in H . somnus genes.

Infect Immun, 1997 Nov, 65(11), 4606 - 14
Use of a novel approach, termed island probing, identifies the Shigella flexneri she pathogenicity island which encodes a homolog of the immunoglobulin A protease-like family of proteins; Rajakumar K et al.; The she gene of Shigella flexneri 2a, which also harbors the internal enterotoxin genes set1A and set1B (F . R . Noriega, GenBank accession no . U35656, 1995) encodes a homolog of the virulence-related immunoglobulin A (IgA) protease-like family of secreted proteins, Tsh, EspC, SepA, and Hap, from an avian pathogenic Escherichia coli, an enteropathogenic E . coli, S . flexneri 5, and Haemophilus influenzae, respectively . To investigate the possibility that this locus was carried on a larger deletable element, the S . flexneri 2a YSH6000T she gene was insertionally disrupted by allelic exchange using a Tn10-derived tetAR(B) cassette . Then, to detect loss of the she locus, the tetracycline-resistant derivative was plated onto fusaric acid medium to select for tetracycline-sensitive revertants, which were observed to arise at a frequency of 10(-5) to 10(-6) . PCR and pulsed-field gel electrophoresis analysis confirmed loss of the she::tetAR(B) locus in six independent tetracycline-sensitive isolates . Sample sequencing over a 25-kb region flanking she identified four insertion sequence-like elements, the group II intron-like sequence Sf.IntA, and the 3' end of a second IgA protease-like homolog, sigA, lying 3.6 kb downstream and in an orientation inverted with respect to she . The deletion was mapped to chromosomal NotI fragment A and determined to have a size of 51 kb . Hybridization with flanking probes confirmed that at least 17.7 kb of the 51-kb deletable element was unique to the seven she+ strains investigated, supporting the conclusion that she lay within a large pathogenicity island . The method described in this study, termed island probing, provides a useful tool to further the study of pathogenicity islands in general . Importantly, this approach could also be of value in constructing safer live attenuated bacterial vaccines.

Infect Immun, 1997 Nov, 65(11), 4488 - 93
Detoxified lipooligosaccharide from nontypeable Haemophilus influenzae conjugated to proteins confers protection against otitis media in chinchillas; Gu XX et al.; Detoxified-lipooligosaccharide (dLOS)-protein conjugates from nontypeable Haemophilus influenzae (NTHi) elicited a significant rise of anti-LOS antibodies with bactericidal activity in rabbits (X.-X . Gu, C.-M . Tsai, T . Ueyama, S . J . Barenkamp, J . B . Robbins, and D . J . Lim, Infect . Immun . 64:4047-4053, 1996) . In this study, we evaluated whether vaccination with the conjugates would protect against NTHi otitis media in chinchillas . Fifty-eight chinchillas received three subcutaneous or intramuscular injections of dLOS-conjugated tetanus toxoid, dLOS-conjugated high-molecular-weight proteins from NTHi, or saline (control) in Freund's adjuvant and then were challenged by intrabullar inoculation with 140 CFU of NTHi . All vaccinated animals responded with elevated serum titers of anti-LOS antibody, and 49% (19 of 39) demonstrated bactericidal activity against the homologous strain . Otitis media with culture-positive NTHi effusions developed in all 19 controls and 56% (22 of 39) of the vaccinated animals during a period of 21 days (P < 0.001) . Bacterial counts of the middle ear effusions were lower in the vaccine groups than in the controls (P < 0.01) . The incidences of infection in the unchallenged ear or inner ear were 26 or 28% in the vaccine groups and 53 or 58% in the controls (P < 0.05) . The signs of infection observed by otoscopy were less severe in the vaccine groups than in the controls . There was no significant difference between the two vaccine groups . These data indicate that active immunization with LOS-based conjugates reduces the incidence of NTHi-induced otitis media.

Infect Immun, 1997 Nov, 65(11), 4452 - 9
Comparative analysis of immunoglobulin A1 protease activity among bacteria representing different genera, species, and strains; Reinholdt J et al.; Immunoglobulin A1 (IgA1) proteases cleaving human IgA1 in the hinge region are produced constitutively by a number of pathogens, including Haemophilus influenzae, Neisseria meningitidis, Neisseria gonorrhoeae, and Streptococcus pneumoniae, as well as by some members of the resident oropharyngeal flora . Whereas IgA1 proteases have been shown to interfere with the functions of IgA antibodies in vitro, the exact role of these enzymes in the relationship of bacteria to a human host capable of responding with enzyme-neutralizing antibodies is not clear . Conceivably, the role of IgA1 proteases may depend on the quantity of IgA1 protease generated as well as on the balance between secreted and cell-associated forms of the enzyme . Therefore, we have compared levels of IgA1 protease activity in cultures of 38 bacterial strains representing different genera and species as well as strains of different pathogenic potential . Wide variation in activity generation rate was found overall and within some species . High activity was not an exclusive property of bacteria with documented pathogenicity . Almost all activity of H . influenzae, N . meningitidis, and N . gonorrhoeae strains was present in the supernatant . In contrast, large proportions of the activity in Streptococcus, Prevotella, and Capnocytophaga species was cell associated at early stationary phase, suggesting that the enzyme may play the role of a surface antigen . Partial release of cell-associated activity occurred during stationary phase . Within some taxa, the degree of activity variation correlated with degree of antigenic diversity of the enzyme as determined previously . This finding may indicate that the variation observed is of biological significance.

Infect Immun, 1997 Nov, 65(11), 4431 - 5
Evaluation of the virulence of nontypeable Haemophilus influenzae lipooligosaccharide htrB and rfaD mutants in the chinchilla model of otitis media; DeMaria TF et al.; Considerable evidence has implicated nontypeable Haemophilus influenzae (NTHi) lipooligosaccharide (LOS) in the pathogenesis of otitis media (OM); however, its exact role has not been conclusively established . Recently, two NTHi LOS-deficient mutants have been created and described . Strain 2019-DK1, an rfaD gene mutant, expresses a truncated LOS consisting of only three deoxy-D-manno-octulosonic acid residues, a single heptose, and lipid A . Strain 2019-B29, an isogenic htrB mutant, possesses an altered oligosaccharide core and an altered lipid A . Each strain's ability to colonize the nasopharynx and to induce OM subsequent to transbullar inoculation was evaluated in the chinchilla model . Nasopharyngeal colonization data indicate that the parent strain and both mutants are able to colonize the nasopharynx and exhibit comparable clearance kinetics . Compared with the parent and each other, however, the mutants demonstrated marked differences in virulence regarding their relative abilities to induce OM and persist in the middle ear post-transbullar inoculation . Strain B29 required a 3-log-greater dose to induce OM than the parent strain and did not exhibit evidence of sustained multiplication but persisted for the same duration as the parent . Conversely, strain-DK1, even when inoculated at a dose 4 logs greater than the parent dose, was eliminated from the middle ear 72 h after challenge . A comparison of the relative pathogenicities of these isolates provides the opportunity to address fundamental questions regarding the contribution of LOS to pathogenesis issues at the molecular level . Specifically, the impact of these LOS gene disruptions on OM pathogenesis can be defined and may thus provide potential new targets for future protection and intervention strategies.

Infect Immun, 1997 Nov, 65(11), 4389 - 94
Genetic and biochemical analyses of Actinobacillus pleuropneumoniae urease; Bosse JT et al.; The urease gene cluster from the virulent Actinobacillus pleuropneumoniae serotype 1 strain CM5 was cloned and sequenced . The urease activity was associated with a 6.3-kbp region which contains eight long open reading frames (ORFs) . The structural genes, ureABC, are separated from the accessory genes, ureEFGD, by a 615-bp ORF of unknown function, ureX . Homologies were found with the structural and accessory urease gene products of Haemophilus influenzae and, to a lesser extent, with those of other organisms . The urease enzyme subunits had predicted molecular masses of 61.0, 11.3, and 11.0 kDa, and the size of the holoenzyme was estimated to be 337 +/- 13 kDa by gel filtration chromatography . Urease activity was maximal but unstable at 65 degrees C . In cell lysates, the A . pleuropneumoniae urease was stable over a broad pH range (5.0 to 10.6) and the optimal pH for activity was 7.7 . The Km was 1.5 +/- 0.1 mM urea when it was assayed at pH 7.7 . The low Km suggests that this enzyme would be active in the respiratory tract environment, where urea levels should be similar to those normally found in pig serum (2 to 7 mM).

J Bacteriol, 1997 Nov, 179(21), 6855 - 7
The acrAB homolog of Haemophilus influenzae codes for a functional multidrug efflux pump; Sanchez L et al.; Disruption of gene HI0894 or HI0895 in Haemophilus influenzae Rd, homologs of Escherichia coli acrAB multidrug efflux genes, caused hypersusceptibility to erythromycin, rifampin, novobiocin, and dyes such as ethidium bromide and crystal violet and increased accumulation of radioactive erythromycin, showing that these genes are expressed and contribute to the baseline level resistance of this organism through active drug efflux . The gene disruption did not produce detectable changes in susceptibility to several other antibiotics, possibly because rapid influx of small antibiotic molecules through the large H . influenzae porin channels counterbalances their efflux.

Trends Genet, 1997 Oct, 13(10), 399 - 404
Haemophilus influence: the impact of whole genome sequencing on microbiology; Tang CM et al.; The publication of the Haemophilus influenzae genome sequence in 1995 was a landmark in microbiological research . It has changed our understanding of the prokaryotic world, and will influence the approach and focus of research on microorganisms over the next few years . In this article we outline what has been learned from this and other genome sequencing projects, and discuss some of the potential avenues of investigation that will follow in the 'post-genome era'.

Acta Clin Belg, 1997, 52(4), 204 - 6
Neonatal sepsis due to nonencapsulated Haemophilus influenzae biotype IV; Van Deynse E et al.; A case report of a newborn with sepsis due to nontypable H.Influenzae biotype IV is presented . There were no prematurity nor maternal obstetrical complications involved . The child however suffered from severe respiratory distress . With the aspiration of secretions, the resuscitation with mask oxygen and the empirically started combination of ampicillin and cefotaxime, his condition rapidly improved.

J Clin Microbiol, 1997 Nov, 35(11), 2854 - 8
Use of multiplex PCR for simultaneous detection of four bacterial species in middle ear effusions; Hendolin PH et al.; A multiplex PCR procedure was developed for the simultaneous detection of Alloiococcus otitidis, Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae in middle ear effusions (MEEs) from patients with chronic otitis media with effusion . The bacterial 16S rRNA gene was chosen as the target, and the procedure used one common lower primer and four species-specific upper primers . The reaction was optimized by changing the primer concentrations to yield equal amounts of amplification products . The specificity of the reaction was verified with various bacterial species found in the nasopharynx . The performance of the procedure was examined with 25 MEE specimens, and the results were compared to those obtained by conventional culture methods . A detection level of 10 bacterial cells/reaction for each of the study organisms was achieved . By conventional culture methods, 8 (32%) of the specimens showed growth of one of the study organisms . In contrast, 21 (84%) of the specimens tested positive by the multiplex PCR . None of the culture-positive specimens were PCR negative, whereas three (12%) of the PCR-positive specimens tested positive for two of the four study organisms . Thus, the multiplex PCR method improves the detection rate significantly compared to that of the conventional culture method.

Diagn Microbiol Infect Dis, 1997 Sep, 29(1), 33 - 8
In vitro activity of trimethoprim alone compared with trimethoprim-sulfamethoxazole and other antimicrobials against bacterial species associated with upper respiratory tract infections; Eliopoulos GM et al.; Trimethoprim-sulfamethoxazole has been used to treat various respiratory tract infections . Nevertheless, for many patients, intolerance of the sulfonamide component precludes use of this combination . This study examined the activity of trimethoprim alone in comparison to that of trimethoprim-sulfamethoxazole and other antimicrobials against bacterial species implicated in respiratory tract infections . For Haemophilus influenzae, minimal inhibitory concentrations of trimethoprim were equal to or one dilution greater than those of trimethoprim-sulfamethoxazole, with 56 of 58 strains inhibited by the former at < or = 0.25 microgram/ml . All oxacillin-susceptible Staphylococcus aureus and 96.7% of Streptococcus pyogenes were inhibited by trimethoprim < or = 2 micrograms/ml . In contrast, only 50% of Streptococcus pneumoniae were inhibited by this concentration of trimethoprim, whereas 93.3% were susceptible to the combination at < or = 2/38 micrograms/ml . All oxacillin-resistant S . aureus and all Moraxella catarrhalis were resistant to trimethoprim, although many of the former and all of the latter were susceptible to trimethoprim-sulfamethoxazole.

Acta Otolaryngol, 1997 Sep, 117(5), 681 - 8
Cochlear changes following destruction of semicircular canal in healthy and previously toxin-exposed rats . An electrophysiological and morphological investigation; Stenqvist M et al.; One group of Sprague-Dawley rats (group A, n = 6) was treated by instilling Pseudomonas aeruginosa exotoxin A (PaExoA), and another (group B, n = 6) treated similarly with Haemophilus influenzae type b endotoxin (HiBEndo) . In group A a 20 dB hearing loss was observed, predominantly in the high-frequency region, which was reversible within 1 month . In group B no significant hearing impairment was noted . Between 1 and 6 months later, the lateral and posterior semicircular canals (SCCs) were ablated unilaterally . Control rats (group C, n = 8) were subjected to ablation only . All rats were cochleotomized contralaterally prior to labyrinthine surgery . Frequency-specific evoked potential testing at 2-31.5 kHz tone bursts was performed before and directly after surgery, 6, 24 and 48 hours and 1, 4 and 16 weeks postoperatively . After surgery in 18 rats, thresholds rose immediately, predominantly at 2, 4 and 6 kHz, followed by varying degrees of recovery . Greatest immediate postoperative hearing loss was observed in group A; no rat recovered completely and two rats showed severe permanent threshold elevation . All group B rats recovered completely, except one showing moderate threshold impairment . No permanent hearing loss was observed in group C . This study shows that destruction of SCCs in rats does not necessarily cause permanent hearing loss, even if the fluid spaces are not sealed off . However, previous exposure of the middle ear to PaExoA (but not HiBEndo) renders the cochlea more vulnerable and can result in persistent hearing loss.

Pediatr Infect Dis J, 1997 Oct, 16(10), 959 - 62
Concentration of cefuroxime in middle ear effusion of children with acute otitis media; Thoroddsen E et al.; BACKGROUND: Antibiotic concentrations in serum and middle ear effusion are important in determining therapeutic success in acute otitis media . For beta-lactams the most relevant pharmacokinetic index for clinical efficacy is the time for which serum concentrations exceed the minimum inhibitory concentration (MIC) of the pathogen, which should be at least 40 to 50% of the dosing interval . METHODS: In this open, single center study, the concentration of cefuroxime achieved in the serum and middle ear effusion of pediatric acute otitis media patients with purulent effusion was assessed between 2 and 5 h after a single oral dose of 15 mg/kg cefuroxime axetil suspension . RESULTS: Serum concentrations of cefuroxime ranged from 2.8 to 7.3 microg/ml and were consistent with the results of previous pharmacokinetic study . These results show that serum concentrations of cefuroxime remain above the MIC90 (2.0 microg/ml) for Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis for at least 5 h (42%) of the 12-h dosing interval . Cefuroxime was detected in 14 of 17 (82%) middle ear effusion samples and ranged from 0.2 to 3.6 microg/ml, indicating that cefuroxime penetrates well into the middle ear . CONCLUSIONS: Cefuroxime is well-absorbed and penetrates well into the middle ear after oral administration of cefuroxime axetil suspension.

Clin Ther, 1997 Jul-Aug, 19(4), 617 - 25; discussion 603
Pharmacologic and clinical comparison of cefaclor in immediate-release capsule and extended-release tablet forms; Cole P; A new controlled-delivery, extended-release 500-mg formulation of cefaclor that is administered twice daily may improve patient compliance compared with the older, immediate-release 250-mg formulation that is administered three times daily . When the extended-release tablet is administered with food, peak plasma cefaclor concentrations are achieved about 2.5 hours after the dose compared with about 1 hour after a dose with the immediate-release capsule . However, the two formulations have an equivalent extent of absorption and equivalent pharmacokinetics after absorption . Cefaclor has retained its excellent in vitro activity against the pathogens most commonly associated with acute exacerbations of chronic bronchitis, namely, Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae . Clinical trials show similar efficacy of the two formulations in patients with acute exacerbations of chronic bronchitis caused by these organisms, but for this indication a 7-day regimen may be used with the extended-release 500-mg formulation compared with a 10-day dosing regimen with the 250-mg capsule . The shorter course of treatment with the new formulation may also improve patient compliancePublication Types:

	Review
	Review, Tutorial

What Is Water Purification?, What Is Genetic Engineering?, What Is Biotechnology?, What Is Listeria Monocytogenes?, What Is Nitrification?, a, Microbe, r, Microbes, c, Microbiology, n, Microorganisms, n, Bacterium, s, Bacillus, r, S. cerevisiae, c, Salmonella typhimurium, n, P. putida, a, Salmonella typhimurium, n, Micrococci, e, Campylobacter, r, Escherichia coli, n, Antimicrobials, c, B. anthracis, o, Botulism, c, Escherichia coli, r, S. cerevisiae, s, Salmonella typhimurium, s, Penicillin, n, Antibiotics, n, Bacillus anthracis, a, Microorganism, a, Multidrug resistant, s, Microorganisms, o, Pasteurella

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