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