Drugs, 1996 Aug, 52(2), 254 - 75 A cellular pertussis vaccine (Infanrix-DTPa; SB-3) . A review of its immunogenicity, protective efficacy and tolerability in the prevention of Bordetella pertussis infection; Patel SS et al.; SB-3 (Infanrix-DTPa) is one of a new generation of vaccines for immunisation against pertussis (whooping cough), diphtheria and tetanus . It is a 3-component (pertussis toxin, filamentous haemagglutinin and pertactin) chemically inactivated acellular pertussis pertussis-diphtheria-tetanus toxoid (DTaP) vaccine, and it differs from conventional whole-cell pertussis-diphtheria-tetanus toxoid (DTwP) vaccines in that it comprises inactivated purified Bordetella pertussis antigens rather than whole cells of the bacillus . SB-3, like a number of other DTaP vaccines, elicits a similar or more often, a significantly greater immune response than various DTwP vaccines in healthy infants and young children . initial data from comparative studies indicate that SB-3 also remains immunogenic when given in combination with hepatitis B vaccine or concurrently administered with Haemophilus influenzae type b (HbOC) conjugate vaccine . A combination of SB-3 and H . influenzae type b tetanus (PRP-T) conjugate vaccine results in lower anti-PRP antibody response than when both vaccines are administered concurrently . Data from two large, multicentre, German and Italian studies in infants indicate that the protective efficacy of SB-3 against pertussis was significantly better than one DTwP (DTwP-CON) but similar to another one (DTwP-BW) under investigation . Compared with another DTaP vaccine (BIO-3), SB-3 was just as protective . Overall, the data from these 2 studies indicate that primary vaccination with SB-3 provides effective protection against pertussis, even under the stringent conditions of a household contact with typical pertussis . As the other DTaP vaccines, SB-3 is better tolerated than DTwP vaccines, with a significantly lower incidence of common adverse events such as local reactions (swelling, pain and a erythema), irritability, fever, persistent crying and local tenderness . Clinical experience with SB-3 thus far indicates that, like other DTaP vaccines, it is associated with significantly fewer common (non-serious) adverse events than DTwP vaccines . Less clear is whether it has any advantage over DTwP vaccines with respect to protective efficacy or over other DTaP vaccines with respect to tolerability and protective efficacy . Nevertheless, the available data support the use of SB-3 for infant immunisation, as well as providing a suitable basis for the development of new combination vaccines. J Clin Microbiol, 1996 Aug, 34(8), 2030 - 2 Detection of Haemophilus influenzae and Streptococcus pneumoniae DNA in blood culture by a single PCR assay; Hassan-King M et al.; A multiplex PCR assay was developed to screen blood cultures from children in The Gambia with suspected pneumonia for the simultaneous detection of Haemophilus influenzae type b and Streptococcus pneumoniae isolates . Analysis of 295 blood cultures showed that PCR detected the organisms in all samples positive by culture in two samples infected with H . influenzae type b and four samples infected with S . pneumoniae that were culture negative, indicating that this method is sensitive for detecting these organisms in blood cultures. J Clin Microbiol, 1996 Aug, 34(8), 1970 - 4 Reevaluation of interpretive criteria for Haemophilus influenzae by using meropenem (10-microgram), imipenem (10-microgram), and ampicillin (2- and 10-microgram) disks; Zerva L et al.; A collection of 300 Haemophilus influenzae clinical strains was used to assess in vitro susceptibility to carbapenems (meropenem, imipenem) by MIC and disk diffusion methods and to compare disk diffusion test results with two potencies of ampicillin disks (2 and 10 micrograms) . The isolates included ampicillin-susceptible or- intermediate (167 strains), beta-lactamase-positive (117 strains), and beta-lactamase-negative ampicillin-resistant (BLNAR; 16 strains) organisms . Disk diffusion testing was performed with 10-micrograms meropenem disks from two manufacturers . Meropenem was highly active against H . influenzae strains (MIC50, 0.06 microgram/ml; MIC90, 0.25 microgram/ml; MIC50 and MIC90, MICs at which 50 and 90%, respectively, of strains are inhibited) and was 8- to 16-fold more potent than imipenem (MIC50, 1 microgram/ml; MIC90, 2 micrograms/ml) . Five non-imipenem-susceptible strains were identified (MIC, 8 micrograms/ml), but the disk diffusion test indicated susceptibility (zone diameters, 18 to 21 mm) . MIC values of meropenem, doxycycline, ceftazidime, and ceftriaxone for BLNAR strains were two- to fourfold greater than those for other strains . The performance of both meropenem disks was comparable and considered acceptable . A single susceptible interpretive zone diameter of > or = 17 mm (MIC, < = or 4 micrograms/ml) was proposed for meropenem . Testing with the 2-micrograms ampicillin disk was preferred because of an excellent correlation between MIC values and zone diameters (r = 0.94) and superior interpretive accuracy with the susceptible criteria at > or = 17 mm (MIC, < or = 1 microgram/ml) and the resistant criteria at < or = 13 mm (MIC, > or = 4 micrograms/ml) . Among the BLNAR strains tested, 81.3% were miscategorized as susceptible or intermediate when the 10-micrograms ampicillin disk was used, while the 2-micrograms disk produced only minor interpretive errors (12.5%) . Use of these criteria for testing H . influenzae against meropenem and ampicillin should maximize reference test and standardized disk diffusion test performance with the Haemophilus Test Medium . The imipenem disk diffusion test appears compromised and should be used with caution for detecting strains for which imipenem MICs are elevated. J Clin Microbiol, 1996 Aug, 34(8), 1926 - 9 Variation in metabolic enzyme activity of persistent Haemophilus influenzae in respiratory tracts of patients with cystic fibrosis; Moller LV et al.; Haemophilus influenzae organisms were isolated from sputum specimens prospectively collected from 40 patients with cystic fibrosis during 2 years to study variations in the metabolic enzyme activities of persistent H . influenzae strains as determined by biotyping . In total, 97 distinct H . influenzae strains without variations in their major outer membrane protein (MOMP) patterns and 73 MOMP variants derived from 30 of these distinct strains were obtained . Twelve distinct strains and 42 MOMP variant strains were isolated at multiple time points during the study period, indicating the persistence of these strains . Among the 54 persistent H . influenzae strains, 22 (41%) strains with stable MOMP compositions showed random variations in biotypes . In 39 of 103 (38%) H . influenzae strains, biotype changes coincided with MOMP variations . Biotype variations were the result of both the loss and the acquisition of enzyme activities . The results of the study indicate that changes in metabolic enzyme activity occur randomly during the persistence of H . influenzae organisms in cystic fibrosis patients, irrespective of MOMP variations. J Ark Med Soc, 1996 Aug, 93(3), 137 - 8 Invasive non-typeable Haemophilus influenzae diseases in children; Schutze GE et al.; The current approach to patients with invasive non-typeable H . influenza disease is based upon past experience with the type b strains . In areas where clinicians cannot obtain typing information in a timely manner, issues concerning treatment and prophylaxis should be approached as if the patients were infected with a type b strain . This approach will not change until further information becomes available on invasive non-typeable H . influenzae infections in children. Infect Immun, 1996 Aug, 64(8), 3161 - 7 Cloning, sequencing, expression, and protective capacity of the oma87 gene encoding the Pasteurella multocida 87-kilodalton outer membrane antigen; Ruffolo CG et al.; Membrane proteins of Pasteurella multocida have been shown previously to elicit protective immunity . We have identified an 87-kDa outer membrane antigen, Oma87, which is present in all 16 serotypes of P . multocida . The gene encoding this protein was cloned and sequenced and found to have significant similarity to the D15 protective surface antigen of Haemophilus influenzae . Oma87 was localized to the outer membrane of the cell, and proteinase K treatment suggested that the protein is surface exposed . Native and recombinant Oma87 were strongly immunostained by convalescent-phase antiserum, indicating that the protein is expressed in vivo . Specific Oma87 antiserum protected mice against homologous, lethal P . multocida challenge . These results suggest that Oma87 is a protective outer membrane antigen of P . multocida. Infect Immun, 1996 Aug, 64(8), 3134 - 41 Cloning of a DNA fragment encoding a heme-repressible hemoglobin-binding outer membrane protein from Haemophilus influenzae; Jin H et al.; Haemophilus influenzae is able to use hemoglobin as a sole source of heme, and heme-repressible hemoglobin binding to the cell surface has been demonstrated . Using an affinity purification methodology, a hemoglobin-binding protein of approximately 120 kDa was isolated from H . influenzae type b strain HI689 grown in heme-restricted but not in heme-replete conditions . The isolated protein was subjected to N-terminal amino acid sequencing, and the derived amino acid sequence was used to design corresponding oligonucleotides . The oligonucleotides were used to probe a Southern blot of EcoRI-digested HI689 genomic DNA . A hybridizing band of approximately 4.2 kb was successfully cloned into pUC19 . Using a 1.9-kb internal BglII fragment of the 4.2-kb clone as a probe, hybridization was seen in both typeable and nontypeable H . influenzae but not in other bacterial species tested . Following partial nucleotide sequencing of the 4.2-kb insert, a putative open reading frame was subcloned into an expression vector . The host Escherichia coli strain in which the cloned fragment was expressed bound biotinylated human hemoglobin, whereas binding of hemoglobin was not detected in E . coli with the vector alone . In conclusion, we hypothesize that the DNA fragment encoding an approximately 120-kDa heme-repressible hemoglobin-binding protein mediates one step in the acquisition of hemoglobin by H . influenzae in vivo. Infect Immun, 1996 Aug, 64(8), 3032 - 7 Identification of surface-exposed B-cell epitopes on high molecular-weight adhesion proteins of nontypeable Haemophilus influenzae; Barenkam SJ et al.; We previously reported that two surface-exposed high-molecular-weight proteins, HMW1 and HMW2, expressed by a prototypic strain of nontypeable Haemophilus influenzae (NTHI) mediate attachment to human epithelial cells . These proteins are members of a family of highly immunogenic proteins common to most nontypeable Haemophilus strains . We also reported that immunization with an HMW1-HMW2 mixture modified the course of disease in an animal model of otitis media, suggesting the potential usefulness of these proteins as NTHI vaccine components . Identification of surface-accessible B-cell epitopes could be important to efforts to develop recombinant or synthetic peptide vaccines based upon these high-molecular-weight proteins . Thus, the purpose of the present study was to identify surface-accessible epitopes on the HMW1 and HMW2 proteins by using monoclonal antibodies (MAbs) and to determine the prevalence of these epitopes among the high-molecular-weight proteins expressed by heterologous nontypeable Haemophilus strains . MAbs were generated by immunizing mice with high-molecular-weight proteins purified from prototype strains and were screened by immunoelectron microscopy (IEM) for the ability to recognize surface epitopes . Two MAbs, designated AD6 and 10C5, that recognized surface epitopes by IEM were recovered . In order to map the epitopes recognized by these two MAbs, we constructed a set of HMW1 and HMW2 recombinant fusion proteins using the pGEMEX vectors and examined the reactivity of the MAbs with these fusion proteins . MAb AD6 recognized an epitope in both HMW1 and HMW2 which mapped to the last 75 amino acids at the carboxy termini of the two proteins . When examined for reactivity with heterologous strains, MAb AD6 recognized high-molecular-weight proteins in 75% of 125 unrelated nontypeable Haemophilus strains and, in addition, reacted with three of three such strains when examined by IEM . MAb 10C5 recognized an epitope that mapped to a 155-amino-acid segment near the carboxy terminus of HMW1 . This epitope was adjacent to but distinct from the AD6 epitope and was absent from HMW2 . The 10C5 epitope was expressed by 40% of the AD6 reactive strains . Identification of shared surface-exposed epitopes on the high-molecular-weight adhesion proteins suggests the possibility of developing recombinant or synthetic peptide-based vaccines protective against disease caused by the majority of NTHI strains. J Allergy Clin Immunol, 1996 Aug, 98(2), 451 - 9 Antibody response to unconjugated Haemophilus influenzae b and pneumococcal polysaccharide vaccines in children with recurrent infections; Raby R et al.; BACKGROUND: Increasingly, antibody testing is being used to evaluate the status of humoral immunity in patients with recurrent infection and suspected immunodeficiency . In the past, we had been impressed that immunization with unconjugated Haemophilus influenzae b (uHib) vaccine provided useful information about the ability to produce antibody to polysaccharides and that the use of pneumococcal polysaccharide (PPS) vaccine frequently produced results that were difficult to interpret . OBJECTIVE: The study was carried out to compare antibody responsiveness to vaccination with uHib with the response seen after PPS vaccination . METHODS: Twenty children (ages, 2 to 13 years; 11 male) who were referred to our immunology clinic because of recurrent infections were immunized with both uHib vaccine and PPS vaccine . Nine children had previously received conjugated Hib vaccine . RESULTS: All 20 children either responded with a twofold or greater increase in antibody titer after uHib vaccine or had preimmunization antibody concentrations of greater than 400 nanograms antibody nitrogen per milliliter (ng Ab N/ml) . All of the children responded to PPS-3 with postimmunization antibody concentrations greater than 400 ng Ab N/ml . Three children had an increase in titer to PPS-7 of less than twofold, seven did not have a twofold increase in titer to PPS-9, and 15 had an increase in titer to PPS-14 of less than twofold . CONCLUSION: Unconjugated Hib vaccine is a potent immunogen in children over 2 years of age . Prior immunization with the conjugate vaccine did not prevent a response to unconjugated vaccine . Unconjugated Hib vaccine appears to be at least as immunogenic as PPS-3 when used as an assessment vaccine for evaluating antibody responsiveness. J Bacteriol, 1996 Aug, 178(15), 4445 - 52 A new gene locus of Bordetella pertussis defines a novel family of prokaryotic transcriptional accessory proteins; Fuchs TM et al.; Recently, a novel type of regulatory mutation causing differential effects on the expression of virulence genes due to a slight overexpression of the RNA polymerase alpha subunit (RpoA) was found in Bordetella pertussis (N . H . Carbonetti, T . M . Fuchs, A . A . Patamawenu, T . J . Irish, H . Deppisch, and R . Gross, J . Bacteriol . 176:7267-7273, 1994) . To gather information on the molecular events behind this phenomenon, we isolated suppressor mutants of the RpoA-overexpressing strains after random mutagenesis . Genetic characterization of these suppressor strains revealed the existence of at least three distinct groups of dominant alleles . Mutations occurred either in the rpoA locus itself, in the bvg locus, or in unknown gene loci . One mutant of the latter group was further characterized . By the introduction of a cosmid library containing genomic B . pertussis DNA into this suppressor strain, we isolated a cosmid which suppressed the phenotype of the suppressor strain, thus restoring the negative effect on transcription of the ptx and cya toxin genes . Mutagenesis of the cosmid with Tn5 led to the identification of the gene locus responsible for this phenomenon . Its DNA sequence revealed the presence of an open reading frame (ORF) consisting of 2,373 bp coding for a hypothetical 86-kDa protein with extensive sequence similarities to ORFs with not yet identified functions of Escherichia coli, Haemophilus influenzae, and Neisseria meningitidis . The new gene, termed tex, for toxin expression, seems to be an essential factor for B . pertussis, as it cannot be deleted from the bacterial chromosome . All members of this new protein family show significant sequence similarities with the mannitol repressor protein MtlR and with the presumptive RNA-binding domains of the Pnp and ribosomal S1 proteins of E . coli in their N- and C-terminal parts, respectively . These sequence similarities and the fact that the tex gene was isolated by virtue of its effects on gene expression in B . pertussis indicate that the members of this new protein family may play an important role in the transcription machinery of prokaryotic organisms. Arch Pediatr Adolesc Med, 1996 Aug, 150(8), 863 - 6 Immunization status as determined by patients' hand-held cards vs medical records; Fierman AH et al.; OBJECTIVE: To determine whether patients' hand-held immunization cards provide accurate assessments of immunization status when compared with their corresponding medical records . SETTING: Urban hospital emergency department immunization program . DESIGN: Comparison of 2 criterion standards . PATIENTS: Children aged 4 months to 6 years who presented consecutively with their immunization cards and received routine care in the hospital's pediatric clinic . SELECTION: Of 673 eligible patients seen in the immunization program from November 1992 to October 1993, 140 were randomly selected for comparison of immunization card and medical record immunization dates; in addition, all 123 eligible patients seen between August and October 1994 were selected . Of the total of 263 children, medical records for 257 (98%) were available for review . The dates of diphtheria-tetanus-pertussis, polio, measles-mumps-rubella, and Haemophilus influenzae type b immunization from immunization cards and medical records were recorded, as were patient age, sex, and ethnicity . Immunization card-medical record immunization date pairs were compared . Each immunization card and medical record was categorized as up to date, due for immunization, or delayed 2 months or more for any immunization at the time of the visit . RESULTS: In 218 (85%) of 257 cases, the immunization card and medical record immunization dates were identical (McNemar test, P = .63) . The immunization card and medical record agreed that patients were due for immunization in 91 cases and agreed that patients were not due for immunization in 138 cases (kappa = 0.77; 95% confidence interval, 0.70-0.85) . The immunization card and medical record agreed that patients were delayed for 1 or more immunizations in 51 cases and agreed that patients were not delayed in 187 cases (kappa = 0.79; 95% confidence interval, 0.71-0.88) . CONCLUSION: The hand-held immunization card is a suitable alternative to the medical record when the need for immunization is assessed or when rates of immunization delay in populations are determined. Arch Pediatr Adolesc Med, 1996 Aug, 150(8), 838 - 41 Response of human immunodeficiency virus-exposed and -infected infants to Haemophilus influenzae type b conjugate vaccine; Rutstein RM et al.; OBJECTIVE: To evaluate the response of human immunodeficiency virus (HIV)-infected and -exposed infants to the primary series and booster dose of Haemophilus influenzae type b (Hib) conjugate vaccine . DESIGN: Retrospective study . PATIENTS AND SETTING: The HIV-exposed and -infected infants who were attending the Special Immunology Family Clinic at The Children's Hospital of Philadelphia (Pa) . MAIN OUTCOME MEASURES: Geometric mean antibody titers (GMTs) to Hib polyribosyl ribitol phosphate capsular antigen were assessed after the primary series and again after the 15-month booster doses . In addition, the percentages of patients who responded with polyribosyl ribitol phosphate antibody levels greater than both 0.15 and 1.0 mg/L were compared between groups . RESULTS: After the 3-dose primary series, the GMTs were lower in the HIV-infected infants compared with those in the HIV-exposed, uninfected infants (0.86 vs 2.30, P = .02) . Forty-six percent of the HIV-infected infants mounted a response ( > 1.0 mg/L) compared with that in 79% of the HIV-exposed infants (P = .05) . Among the HIV-infected infants, there was no difference in the GMTs based on CD4+ cell counts or HIV-related symptoms . After the 15-month booster dose, the GMTs were not significantly different in the HIV-infected and -exposed infants . As a group, the HIV-infected infants responded to the booster dose with a 2-fold increase in the GMTs, and significantly more of these infants had antibody concentrations above 1.0 mg/L compared with their response to the primary series (62% vs 38%, P = .02) . CONCLUSIONS: Most of the HIV-infected infants responded to the primary series of Hib conjugate vaccine with antibody concentrations greater than 0.15 mg/L, but the GMTs were significantly lower than those in the uninfected infants . The primary series of Hib conjugate vaccine appeared to be capable of inducing specific immunologic memory in the HIV-infected infants . The HIV-infected infants had a significant response to a booster dose of Hib conjugate vaccine, as measured by using the GMTs and the percentage of infants with antibody concentrations greater than 1.0 mg/L . The duration of protective titers will need to be followed in this population of patients who are at a high risk for serious bacterial disease. J Infect Dis, 1996 Aug, 174(2), 427 - 30 An immunohistochemical analysis of naturally occurring chancroid; King R et al.; Haemophilus ducreyi is a major cause of genital ulcer disease in many developing countries and is associated with augmented transmission of human immunodeficiency virus (HIV) . However, the mechanisms through which H . ducreyi produces ulceration are poorly understood . The characteristics of the host response to H . ducreyi and the pathobiology of its potential contribution to increased HIV susceptibility are not known . Chancroid ulcer biopsies from 8 patients were analyzed histologically and immunohistochemically . All biopsies had perivascular and interstitial mononuclear cell infiltrates that extended deep into the dermis . The infiltrate, which contained macrophages and CD4 and CD8 lymphocytes, was consistent with a delayed hypersensitivity type cell-mediated immune response . The recruitment of CD4 T lymphocytes and macrophages may in part explain the facilitation of HIV transmission in patients with chancroid. Arch Ophthalmol, 1996 Aug, 114(8), 943 - 9 Endophthalmitis after filtering surgery with mitomycin; Greenfield DS et al.; OBJECTIVE: To identify the incidence, causative organisms, and clinical outcomes of eyes with bleb-associated endophthalmitis after glaucoma filtering procedures with adjunctive mitomycin . METHODS: Retrospective analysis of 773 consecutive eyes that underwent glaucoma filtering surgery at the Bascom Palmer Eye Institute, Miami, Fla . The course of 609 eyes from 485 patients with a minimum of 3 months of follow-up were reviewed . RESULTS: Mean follow-up was 16.0 +/- 11.5 months (range, 3-48 months) . Of the 609 eyes, 13 (2.1%) developed bleb-associated endophthalmitis an average of 18.5 +/- 13.2 months after surgery (range, 1-45 months) . The incidence of bleb-associated endophthalmitis was significantly greater after inferior trabeculectomy (7.8% per patient-year) than after superior trabeculectomy (1.3% per patient-year) by Kaplan-Meier estimates (P = .02, log rank test) . The cumulative incidence was 13% for inferior limbal blebs and 1.6% for superior limbal blebs . Nine (69.2%) of the 13 eyes were culture positive . Streptococcus sanguis and Haemophilus influenzae (6/13 {46.2%}) were the most frequent causative organisms . The mean increase in intraocular pressure after endophthalmitis treatment was 1.2 mm Hg, with a mean decrease in visual acuity of 1.42 logMAR units . Eight (61.5%) of the 13 eyes had a final acuity of 20/400 or better . CONCLUSIONS: The incidence of bleb-associated endophthalmitis after guarded filtering surgery performed with adjunctive mitomycin is higher than the reported rate in eyes undergoing filtering surgery without the use of antifibrotic agents (0.2%-1.5%) . Inferior limbal trabeculectomy carries the highest risk of infection . Eyes with mitomycin blebs maintained excellent filtration capacity . However, after treatment of the infection, the visual outcomes were generally poor. FEMS Microbiol Lett, 1996 Jul 15, 141(1), 89 - 95 Identification of the pdxK gene that encodes pyridoxine (vitamin B6) kinase in Escherichia coli K-12; Yang Y et al.; We isolated a miniTn10(Cmr) insertion mutant lacking pyridoxine (PN) kinase and cloned the structural gene, designated pdxK, by complementation . P1 transduction and PCR mapping and DNA sequence analysis showed that pdxK was adjacent to the crr sugar transport gene (53.95 min) . Growth properties of pdxK::miniTn10 mutants supported the hypotheses that PN kinase, which also phosphorylates pyridoxal (PL) and pyridoxamine (PM) in vitro, functions solely in the B6-vitamer salvage pathway and that E . coli contains an additional PL kinase . The amino acid sequence of PdxK has signature motifs of the PfkB superfamily of carbohydrate kinases, which includes phosphofructokinases and ribokinases, and suggests that three unidentified ORFs of Salmonella typhimurium, Haemophilus influenzae, and Saccharomyces cerevisiae correspond to PN/PL/PM kinases. Nucleic Acids Res, 1996 Jul 15, 24(14), 2760 - 6 Cloning and analysis of the genes encoding the type IIS restriction-modification system HphI from Haemophilus parahaemolyticus; Lubys A et al.; The genomic region encoding the type IIS restriction-modification (R-M) system HphI (enzymes recognizing the asymmetric sequence 5'-GGTGA-3'/5'-TCACC-3') from Haemophilus parahaemolyticus were cloned into Escherichia coli and sequenced . Sequence analysis of the R-M HphI system revealed three adjacent genes aligned in the same orientation: a cytosine 5 methyltransferase (gene hphIMC), an adenine N6 methyltransferase (hphIMA) and the HphI restriction endonuclease (gene hphIR) . Either methyltransferase is capable of protecting plasmid DNA in vivo against the action of the cognate restriction endonuclease . hphIMA methylation renders plasmid DNA resistant to R.Hindill at overlapping sites, suggesting that the adenine methyltransferase modifies the 3'-terminal A residue on the GGTGA strand . Strong homology was found between the N-terminal part of the m6A methyltransferasease and an unidentified reading frame interrupted by an incomplete gaIE gene of Neisseria meningitidis . The HphI R-M genes are flanked by a copy of a 56 bp direct nucleotide repeat on each side . Similar sequences have also been identified in the non-coding regions of H.influenzae Rd DNA . Possible involvement of the repeat sequences in the mobility of the HphI R-M system is discussed. Riv Eur Sci Med Farmacol, 1996 Jul-Aug, 18(4), 163 - 7 Hemophilus influenzae type b meningitis: pediatric overview; Catania S et al.; The authors valued the incidence and clinical therapeutic aspects of Haemophilus influenzae type b (Hib) meningitis in children . They report a retrospective study, in children, with diagnosis of acute purulent meningitis, from January 1982 to December 1994, aged between 1 month and 14 years . Particular attention was direct to Haemophilus influenzae type b meningitis (20 cases) . The incidence rate of Hib meningitis in the overall cases (89) was 22.47% (20), while among children younger than 5 years Hib was the most frequently pathogen isolated (20/58-34.47%) . In 1/4 of cases, particularly in children younger than 1 years, exordium was aspecific and unclear . At admission culture and examination of Cerebrospinal Fluid (CFS) have been done . CFS was cultured on blood agar and chocolate plates . A latex agglutination test was used for rapid detection of the bacterial antigens . In some cases we looked for bacterial antigens in urine . 20% of children had complications and 10% had sequelae (1 years of follow-up) . We didn't have any dead . Antibiotic treatment was principally with Ampicillin, Cephalosporin and Chloramphenicol . The results of this study confirm the Hib gravity and suggest that the administration of conjugate vaccine against Hib to all living in Italy is justified. Bacteriol Virusol Parazitol Epidemiol, 1996 Jul-Dec, 41(3-4), 135 - 40 {Pharyngitis produced by Arcanobacterium haemolyticum}; Dorobat O et al.; Arcanobacterium haemolyticum (A.h.) was recovered from 0.43% of throat cultures of 3715 patients with sore throat, scarlatina and various skin rash . In a 57.90% patients A.h . was the only bacterial pathogen isolated, while in the remainder beta-haemolytic streptococci, Haemophilus and S . aureus were also detected . beta-haemolytic streptococci were much more frequent than A.h . in the throat cultures (26.83%) . All A.h . strains were of the smooth type based on colony morphology and the ability to ferment sucrose . The strains were susceptible to penicillins, cephalosporins, erytromycin and resistant to trimethoprim-sulphamethoxazole . Pharyngeal injection in 89.47% and exanthem in 78.90% were the most common signs present to the patients. Rev Argent Microbiol, 1996 Jul-Sep, 28(3), 111 - 7 {Utilization of different microbiological markers in the study of Haemophilus influenzae}; Azahares Romero LE et al.; The present study includes 178 Haemophilus influenzae strains isolated in different pediatric hospitals from Havana, Cuba, during 1991-1994, associated to divers infections (meningitis, respiratory sepsis, primary bacteremia) . A combination of various typing and subtyping methods was used as epidemiological markers: serotyping (slide agglutination with diagnostical serum a-f and latex agglutination), biotyping according to Killian's procedures (by determination of indole production, urease and ornithine decarboxylase activity), subtyping by fermentative profiles according to Roberts' methods (glucose, maltose, xylose and fructose) and outer membrane protein profile subtyping (vesicles extraction by a modified Barenkamp's method, analysis by lineal and gradient SDS-PAGE and assessment according to our own classification system) . Serotype b was identified in 89.3%, biotype I was the most frequent (79.1%), other biotypes (II, III, IV and V) were also identified . Fermentative profile D (glucose, maltose, xylose and fructose positive) was the most frequent (52.8%) while profile G (glucose, maltose, xylose positive and fructose negative) represented 20.2% . Other known profiles were present . PA2 (33.7%) was the most frequent OMP subtype . Even though 11 different protein subtypes were found, the 77.5% of the strains were located in only three OMP electrophoretic subtypes (PA2, PC1, LA2). Emerg Infect Dis, 1996 Jul-Sep, 2(3), 176 - 82 Conjugate vaccines and the carriage of Haemophilus influenzae type b; Barbour ML; Pharyngeal carriage of Haemophilus influenzae type b (Hib) is important in the transmission of Hib organisms, the pathogenesis of Hib disease, and the development of immunity to the bacterium . The remarkable success of current vaccination programs against Hib has been due in part to the effect of conjugate Hib vaccines in decreasing carriage of Hib . This review explores evidence for this effect, and discusses the possible mechanisms of the mucosal influence of Hib conjugate vaccines. Int J STD AIDS, 1996 Jul, 7(4), 269 - 75 Sexually transmitted diseases: a survey of case management in Malawi; Chilongozi DA et al.; A national survey of sexually transmitted disease (STD) case management was carried out at 39 health care facilities in Malawi in 1994 . Fifty-four health care providers were observed managing 150 patients presenting with selected STD syndromes and 103 providers were interviewed . STD case management was assessed by calculation of WHO/GPA prevention indicators (PIs) from observation data . The overall rate for PI-6, which measures correct assessment and treatment of STD patients was 11% (81% for history taking, 46% in physical examination, and 13% correct antibiotic treatment according to national guidelines) . The score for PI-7, which measures overall patient counselling was 29% (65% for partner notification and 40% for condom advice) . Although Haemophilus ducreyi is at least as common as Treponema pallidum as the causative agent for genital ulcers, only 16% of patients with genital ulcers were treated effectively for chancroid vs 56% for syphilis . Female patients received less comprehensive care than male STD patients . Only 20% of STD patients were offered condoms . Overall, the survey results support the policy decision to adopt syndromic management of STDs, and provide baseline information for planning and evaluation of a national control programme. Eur J Clin Microbiol Infect Dis, 1996 Jul, 15(7), 556 - 60 Prospective study of epidemiology and prognostic factors in community-acquired pneumonia; Gomez J et al.; Of 342 patients with community-acquired pneumonia, 100 were diagnosed etiologically . In these patients, disease epidemiology, prognostic factors, and influence of antibiotic treatment were analyzed prospectively . Fifty-two patients were treated with a broad-spectrum antibiotic (ceftriaxone), and 48 received a medium-spectrum antibiotic (cefuroxime); some patients in each group also received erythromycin . Streptococcus pneumoniae was the most frequently isolated microorganism (43%), followed by Chlamydia pneumoniae (21%), Haemophilus influenzae (19%), and Mycoplasma pneumoniae (11%) . Factors significantly associated with increased mortality were initially critical or poor clinical condition, involvement of two or more lobules, and complications . Prior administration of antibiotics was predictive of penicillin and erythromycin resistance in Streptococcus pneumoniae, but had no effect on the course of the disease . Eight patients died, 89 were cured, and three had recurrences; there was no significant difference in outcome between treatment groups, regardless of whether patients also received erythromycin . Increased knowledge of epidemiological, predictive, and prognostic factors can significantly improve early diagnosis of community-acquired pneumonia and facilitate the choice of appropriate antibiotic treatment, thereby helping to reduce morbidity and mortality. J Antimicrob Chemother, 1996 Jul, 38 Suppl A, 155 - 70 The Alexander Project: using in-vitro susceptibility data for choosing empirical therapy in LRTI; Gruneberg RN; An international collaborative survey of susceptibility in community-acquired lower respiratory tract infection pathogens collected > 6000 strains from six countries during 1992 and 1993 . The four major pathogens were Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Staphylococcus aureus . MICs of 15 antibiotics were determined and sensitivity interpretations applied using breakpoints based on those of the NCCLS . This analysis highlighted some anomalies, notably for beta-lactams against S . pneumoniae and macrolides against H . influenzae, where apparent sensitivity proportions did not accord with the distribution of MICs . Further analyses were undertaken in order to rank the antibiotics in order of potential usefulness for empirical treatment of LRTI: these included in-vitro potency (mode MIC and MIC90) and a pharmacodynamic comparison, using the ratio Cmax (free drug): MIC90 . According to study breakpoints, the most active agents overall were, for S . pneumoniae, cefuroxime, clarithromycin, ofloxacin and chloramphenicol; for H . influenzae, azithromycin, amoxycillin/ clavulanate, cefixime, ceftriaxone, quinolones and doxycycline . However, other analyses suggested that the most active agents overall were, for S . pneumoniae, amoxycillin (+/- clavulanate) and ceftriaxone, and, for H . influenzae, quinolones, ceftriaxone, cefixime and amoxycillin/clavulanate . Overall, the antimicrobial agents with the greatest potential usefulness for empirical treatment were amoxycillin/ clavulanate, ceftriaxone, cefuroxime, ofloxacin and co-trimoxazole . The choice of empirical therapy depends upon local epidemiology and clinician choice, but the Project data may be of value in the decision-making process. J Antimicrob Chemother, 1996 Jul, 38 Suppl A, 141 - 54 Relevance of the Alexander Project: pharmacodynamic considerations; Drusano GL et al.; Application of pharmacodynamic principles for interpretation of data generated by the Alexander Project is possible for beta-lactam, quinolone and macrolide antibiotics . For beta-lactams, the time that serum concentrations remain above the MIC of the pathogen (T > MIC) is the parameter most closely linked with outcome . It has been shown that T > MIC need be only 50-60% of a dose interval . Since the MIC has the greatest influence on this parameter, a conservative estimate of activity would use the MIC90 . The only beta-lactam antibiotics in the Alexander Project for which T > MIC90 for the four major pathogens (Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Staphylococcus aureus) exceeded 50% of the dose interval were amoxycillin/clavulanate (500/125 mg) and ceftriaxone . For macrolides, T > MIC is relevant for erythromycin and clarithromycin, but not azithromycin, for which AUC is the parameter most closely linked to outcome . Erythromycin, clarithromycin and azithromycin showed efficacy against M . catarrhalis only at MIC90 . Quinolones (ciprofloxacin and ofloxacin), for which AUC is also the relevant pharmacodynamic parameter, had the greatest activity against H . influenzae and M . catarrhalis at MIC90, but were less effective against S . pneumoniae and S . aureus . Susceptibility data such as those provided by the Alexander Project can aid clinicians in choosing appropriate treatment for LRTI based on pharmacodynamic principles. J Antimicrob Chemother, 1996 Jul, 38 Suppl A, 133 - 40 The clinical relevance of in-vitro resistance to penicillin, ampicillin, amoxycillin and alternative agents, for the treatment of community-acquired pneumonia caused by Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis; Klugman KP; The documentation of antimicrobial resistance in respiratory pathogens, contained within the Alexander Project, does not necessarily translate into clinical resistance in the treatment of primary community-acquired pneumonia . There is, in particular, little evidence that penicillin resistance in pneumococci is clinically relevant for the treatment of pneumonia, and there is further evidence that the production of beta-lactamase by Haemophilus influenzae may not always be clinically relevant within this setting . beta-Lactamase producing H . influenzae and Moraxella catarrhalis should probably be treated with alternative agents when they cause exacerbations of chronic bronchitis . More studies are required to define the clinical breakpoints of macrolide and co-trimoxazole resistance in the treatment of pneumonia. J Antimicrob Chemother, 1996 Jul, 38 Suppl A, 117 - 32 Trends in antibiotic resistance of respiratory pathogens: an analysis and commentary on a collaborative surveillance study; Baquero F; The evolution of antibiotic resistance was studied among common respiratory tract pathogens in five countries of the European Union and in the USA during 1992-1993 . The data obtained from a collaborative surveillance study were submitted to population analysis, to detect possible shifts in antibiotic susceptibility and, therefore, associated mechanisms of resistance . Among the emerging haemophilus influenzae phenotypes were isolates that did not correspond to the beta-lactamase negative, amino-penicillin resistant (BLNAR) phenotype, but were beta-lactamase producers showing low level ceftriaxone resistance (early extended spectrum beta-lactamases?) amoxycillin susceptible strains with low level ceftriaxone resistance (PBP modification?) and isolates with high-level fluoroquinolone resistance . Moraxella catarrhalis resistance to ceftriaxone erythromycin or fluoroquinolones was noted . The quantitative evolution of antibiotic resistance may reach saturation in some countries with a very high proportion of resistant strains, for example, Spain and France . Qualitatively, resistant strains may be selected that have broader or more effective mechanisms of resistance, particularly under the recently introduced pressure of more active antibiotics of the same family . In countries with modest levels of antibiotic resistance (UK, Germany, Italy), attention should be paid to the misuse of antibiotics with a propensity to select low-level resistant strains . In this respect, the relative prescribing of aminopenicillins and oral cephalosporins in the UK (a high ratio and low prevalence of Streptococcus pneumoniae) and resistance to penicillin in the USA (a low ratio and high prevalence of resistance) is of potential importance. J Antimicrob Chemother, 1996 Jul, 38 Suppl A, 59 - 69 Antimicrobial resistance among lower respiratory tract isolates of Haemophilus influenzae: results of a 1992-93 western Europe and USA collaborative surveillance study . The Alexander Project Collaborative Group; Doern GV; During 1992 and 1993, 2718 respiratory tract isolates of Haemophilus influenzae were obtained from two study centres in each of five West European countries and five study centres in the USA . beta-Lactamase production was assessed and MICs of 14 antimicrobial agents determined in a single co-ordinating laboratory using a broth microdilution method in Mueller-Hinton-lysed horse blood medium . The prevalence of strains producing beta-lactamase varied between 0 and 37.9% . In general, the highest prevalence was in study centres from Spain and the USA with slightly lower rates observed in France and the UK . Only a single confirmed beta-lactamase-negative, ampicillin resistant strain was recovered during the entire study . Erythromycin resistance, defined as an MIC of > or = 4.0 mg/L, was noted in 57.5% of isolates . Among the other antimicrobials tested, resistance rates > or = 1.0% were observed only with cefaclor (3.7%), chloramphenicol (1.4%) and co-trimoxazole (2.5%) . In no case, was the prevalence of resistance or beta-lactamase production significantly greater in 1993 than in 1992. J Antimicrob Chemother, 1996 Jul, 38 Suppl A, 1 - 57 A multicentre collaborative study of the antimicrobial susceptibility of community-acquired, lower respiratory tract pathogens 1992-1993: the Alexander Project; Felmingham D et al.; The Alexander Project is a unique, international, collaborative antimicrobial susceptibility surveillance study of bacterial pathogens causing community-acquired lower respiratory tract infection . Fifteen centres, ten in the European Union (EU) and five in the USA, each submitted up to 400 isolated per year for 2 years (1992 and 1993) to a central laboratory for re-identification and determination of MICs of 15 antimicrobials using the Sensititre microbroth incorporation technique . Of the total of 6385 isolates collected, Haemophilus influenzae (2718), Streptococcus pneumoniae (1856) and Moraxella catarrhalis (818) were the most frequently identified pathogens . Staphylococcus aureus (690) . Haemophilus parainfluenzae (183) and Klebsiella pneumoniae (120) were identified less commonly . High-level penicillin resistance in S . pneumoniae (MIC > or = 2 mg/L) was found in 222 isolates, an overall prevalence of 12% which varied from < 1% in Germany, Italy, UK and two of the five USA centres, to 3.8-40.4% in the remainder, with the highest prevalence found in France and Spain . Intermediate penicillin resistance (MIC 0.12-1 mg/L) was identified in 228 isolates of S . pneumoniae, an overall prevalence of 12.3%, with individual centre prevalence varying widely (EU, 0-52.3%; USA, 0-20.9%) and not always following that of high-level resistance . Resistance to other, unrelated, antimicrobials, except notably the fluoroquinolones, was strongly associated with beta-lactam resistance . beta-lactamase production was detected in 492 isolates of H . influenzae, an overall prevalence of 18.1% . Rates of detection varied widely between centres from 1.4% in Weingarten, Germany in 1993 to 38.5% in Barcelona, Spain in 1992 . In general, the prevalence of beta-lactamase production was higher and less variable in USA centres than in those of the EU . beta-Lactamase was detected consistently in the majority of isolates of M . catarrhalis with an overall prevalence of 81.7% . Virtually no other resistance phenotype was recognised in this species . Of the 690 collected, most isolates of S . aureus produced beta-lactamase with rates of detection varying from 52.2%-89.1% . Isolates from two centres, Genoa, Italy in 1992 and Paris, France in 1993, were associated with a high prevalence of methicillin-resistance (34.8% and 43.8%, respectively) . Combined isolates of H . parainfluenzae and K . pneumoniae accounted for only 4.7% of the total collection . Although the current data are insufficient to allow analysis of trends in resistance, the study participants have continued to collect further isolates in 1994 and 1995 which will be reported in the future. J Vet Diagn Invest, 1996 Jul, 8(3), 332 - 6 In vitro activity of ceftiofur and its primary metabolite, desfuroylceftiofur, against organisms of veterinary importance; Salmon SA et al.; Ceftiofur (XNL) and its primary metabolite, desfuroylceftiofur (DXNL), were evaluated for in vitro activity against 539 isolates from veterinary sources . Actinobacillus pleuropneumoniae, Pasteurella spp., Haemophilus somnus, Salmonella spp., Escherichia coli, staphylococci, and streptococci were tested . Overall, XNL and DXNL were equivalent in activity against the gram-negative organisms with all minimum inhibitory concentrations (MICs) within 1 serial dilution . Against the staphylococci, MIC difference of 2-3 serial dilutions were detected with an MIC90 for XNL and DXNL of 1.0 and 4.0-8.0 micrograms/ml, respectively . Although the MIC90 obtained for Streptococcus suis for each compound was within 1 dilution, the MIC values against individual strains were 2-3 dilutions greater for DXNL than for XNL . The MICs obtained with the bovine and equine streptococci for DXNL (MIC90 = 0.03 microgram/ml) were 5 serial dilutions higher than those obtained for XNL (MIC90 < or = 0.0019) . Although DXNL was less active than XNL against the streptococci, these differences were not clinically important because both XNL and DXNL were highly active for these bacteria . Although these differences are of little importance with the streptococci, they may have important implications for susceptibility testing of the staphylococci . In conclusion, with the exception of the staphylococci, both XNL and DXNL were highly active against the organisms tested, with MICs for both compounds several fold lower than plasma levels achieved during dosing of XNL. Mol Microbiol, 1996 Jul, 21(1), 147 - 58 Purification and characterization of the integrase from the Haemophilus influenzae bacteriophage HP1; identification of a four-stranded intermediate and the order of strand exchange; Hakimi JM et al.; The integrase encoded by the temperate phage HP1 promotes the site-specific recombination between DNA sites on its genome (the attP site) and on the genome of the host Haemophilus influenzae (the attB site) . The protein has been overproduced in Escherichia coli, and purified to apparent homogeneity . HP1 integrase promotes recombination of supercoiled attP-containing molecules with linear segments with attB sites . Reaction was enhanced by spermidine and by the bacterial DNA-bending protein integration host factor . The rate of recombination showed complex and related dependence upon the integrase concentration and the concentration of the supercoiled attP substrate . These relationships probably originate from the need to assemble a multi-protein complex on the attP DNA . The reaction promoted by HP1 integrase produced a four-stranded initial reaction product in which one pair of DNA strands had undergone transfer while the other pair remained intact . This four-stranded component was produced more rapidly than any product, and its steady-state level was proportional to the overall rate of reaction . This component had the kinetic and structural properties of an intermediate in the recombination reaction . The existence of this intermediate was used to determine that the two strand exchanges required for recombination of the duplex substrates proceed in a defined order. Mol Microbiol, 1996 Jul, 21(1), 21 - 31 Influence of pili, fibrils, and capsule on in vitro adherence by Haemophilus influenzae type b; St Geme JW 3rd et al.; Haemophilus influenzae type b is an encapsulated bacterium that initiates infection by colonizing the upper respiratory epithelium . In vitro studies indicate that H . influenzae type b is capable of expressing two morphologically distinct filamentous adhesive structures, referred to as pili and fibrils, respectively . In this study, we examined adherence to a variety of human epithelial-cell types and demonstrated that pili and fibrils have separate cellular binding specificities . In addition, we found that capsular material inhibits fibril recognition of the host-cell surface . This inhibitory effect was reduced when bacteria were grown to stationary phase, reflecting diminished encapsulation . However, when growth medium was supplemented with Mg2+, stationary-phase organisms were relatively heavily encapsulated and non-adherent . These observations suggest that encapsulation can be modulated in response to growth phase or environmental signals . It is possible that encapsulation is down-modulated early in the infectious process in order to avoid interfering with colonization . In contrast, encapsulation may be up-modulated between hosts and during bacteremia, where it appears to confer a selective advantage . We speculate that this model may also apply to other encapsulated pathogens. Mol Microbiol, 1996 Jul, 21(1), 13 - 9 An isogenic haemolysin-deficient mutant of Haemophilus ducreyi lacks the ability to produce cytopathic effects on human foreskin fibroblasts; Palmer KL et al.; The haemolysin of Haemophilus ducreyi is the newest member of the Proteus/Serratia family of pore-forming toxins . In order to assess the role of the haemolysin in virulence, we constructed an isogenic haemolysin-deficient mutant of H . ducreyi strain 35000 This strain, designated 35000-3, lacks detectable haemolytic activity . We tested H . ducreyi strains 35000 and 35000-3 for their cytopathic activity against human foreskin fibroblasts (HFFs) . We observed strong cytopathic activity when strain 35000 was co-cultured with HFFs . In contrast, cytopathic activity was not observed when strain 35000-3 was co-cultured with HFF cells . We also analysed the isogenic pair of H . ducreyi strains for cytopathic activity against HeLa cells and the keratinocyte cell line HaCaT . Strains 35000 and 35000-3 were strongly cytotoxic when co-cultured with HeLa cells . HaCaT monolayers were slightly damaged by cocultivation with strain 35000-3 but this damage was much less than that observed when HaCaT cells were cocultured with strain 35000 . These results indicate that the H . ducreyi haemolysin is responsible for the previously observed cytotoxic activity against HFF cells and is partially responsible for the activity observed with HaCaT cells . The haemolysin, however, is not responsible for the activity observed with HeLa cells. CLAO J, 1996 Jul, 22(3), 213 - 4 Haemophilus influenzae central corneal ulcer associated with cosmetic lens wear; Leahey AB et al.; PURPOSE: We report a case of keratitis caused by Haemophilus influenzae involving daily wear contact lens use . METHODS: After positive cultures from corneal scrapings and contact lenses, the patient was diagnosed with Haemophilus influenzae keratitis . RESULTS: After 16 days of treatment with topical ciprofloxacin and trimethoprim sulfate-polymyxin B sulfate, the keratitis resolved . CONCLUSIONS: Cultures will help identify rare corneal pathogens such as Haemophilus influenzae and help in directing appropriate treatment. Vet Microbiol, 1996 Jul, 51(1-2), 95 - 104 Evaluation of a PCR for detection of Actinobacillus pleuropneumoniae in mixed bacterial cultures from tonsils; Gram T et al.; A PCR for the detection of Actinobacillus pleuropneumoniae was evaluated . All of 102 field, isolates of A, pleuropneumoniae reacted in the PCR by amplification of a 985 bp product . No PCR amplification product was observed when examining strains of A . ureae, A . capsulatus, A . hominis, A . equuli, A, rossii, A . suis, Escherichia coli, Bordetella bronchiseptica . Streptococcus suis, Pasteurella haemolytica, Pasteurella multocida, Haemophilus parasuis, Haemophilus taxon Minor group, Haemophilus taxon D/E and haemophilus taxon F . Amplification of a 985 bp product was, however, observed when testing strains of A . lignieresii . The lower detection limit of the PCR test was 10(3) A . pleuropneumoniae CFU/PCR test tube and was not affected by addition of 10(6) E . coli CFU/PCR test tube . Mixed bacterial cultures from tonsils of 101 pigs from 9 different herds were tested by culture and by PCR using four different bacteriological media . While 65% reacted positive in the PCR only 23% were positive by culture, thereby suggesting a superior sensitivity of the PCR test to that of culture . The use of selective media, large inoculum and incubation for 48 h gave the highest number of positive PCR reactions from mixed bacterial cultures . Tonsil cultures from 50 pigs from an A . pleuropneumoniae-negative herd did not react in the PCR . The results show that PCR on mixed bacterial cultures from tonsils may be a highly sensitive method for the detection of A . pleuropneumoniae in pig herds. Turk J Pediatr, 1996 Jul-Sep, 38(3), 289 - 93 Incidence of H . influenzae in a day-care center; Akcakaya N et al.; In this study nasopharyngeal haemophilus influenzae flora of healthy children in a day-care center in istanbul were analyzed . Nasopharyngeal cultures of 168 children between two and five years of age were obtained between December 1, 1992 and April 1, 1993 and investigated . H . influenzae was isolated in 104 cultures . H . influenzae type b (Hib), type f and H . parainfluenzae were found 87 children (51.8%), 15 children (8.9%) and one child (0.6%), respectively, while non-typable H . influenzae was discovered in one child (0.6%) . Hib, which is the cause of invasive H . influenzae infection in childhood, was evaluated with respect to age; its incidence was found to be highest in two and three-year-old children, and reduced in children older than four years of age . Although Hib was seen in 518 percent of normal children in the day-care center, invasive Hib disease was not seen in any of those children . Therefore, these children have considered carrier of Hib without clinical manifestations. Clin Infect Dis, 1996 Jul, 23(1), 107 - 13 Community-acquired pneumonia in a cohort of former injection drug users with and without human immunodeficiency virus infection: incidence, etiologies, and clinical aspects; Boschini A et al.; Although the association among bacterial pneumonia, human immunodeficiency virus (HIV) infection, and injection-drug use seems to have been well established, accurate estimates of the risk of community-acquired pneumonia among HIV-positive and HIV-negative injection-drug users (IDUs) are still needed . To estimate the incidence of pneumonia in a community of former IDUs, we followed 4,236 persons between 1991 and 1994; 1,114 (26.3%) were HIV-positive and 3,122 (73.7%) were HIV-negative . All patients were evaluated for pneumonia by standard criteria, a serum sample was obtained from each participant at least once a year, and laboratory values were monitored . Overall, 149 episodes of pneumonia occurred among HIV-positive patients and 61 among HIV-negative patients; incidence rates were 90.5 and 14.2 (per 1,000 person-years), respectively . The most common etiologic agents were Streptococcus pneumoniae, Chlamydia pneumoniae, and Haemophilus influenzae . Among the HIV-positive former IDUs, there was a 1.37-fold increase in the relative risk of pneumonia for every decrease of 100/mm3 in the CD4 cell count (95% confidence interval, 1.16-1.61) . The incidence of community-acquired pneumonia was markedly higher among HIV-positive participants than among HIV-negative ones, a finding similar to that concerning the general population. Nihon Kyobu Shikkan Gakkai Zasshi, 1996 Jul, 34(7), 759 - 64 {Prospective study of the etiology of community-acquired pneumonia among patients in a general hospital}; Ishida T et al.; We prospectively studied the etiology of community-acquired pneumonia among all patients who were admitted to our hospital from July 1994 to June 1995 . Tests for microbial pathogens including Chlamydia spp . and Legionella spp . were performed and diagnoses were made with strict criteria . A total 110 patients with 111 episodes of pneumonia were evaluated, and a pathogen was identified in 61 episodes (55%) . The most common pathogen was Streptococcus pneumoniae (18%), followed by Haemophilus influenzae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycoplasma pneumoniae, and Chlamydia spp . Infection with Legionella pneumophila was not found . Dual pathogens were identified in five episodes . Few prospective studies of the etiology of community-acquired pneumonia have been done in Japan . To prepare guidelines for the management of community-acquired pneumonia in Japan, a national study of the etiology of pneumonia is necessary. Antimicrob Agents Chemother, 1996 Jul, 40(7), 1741 - 4 Ciprofloxacin-resistant Haemophilus influenzae strains possess mutations in analogous positions of GyrA and ParC; Georgiou M et al.; The nucleotide sequences of the quinolone resistance-determining regions of the gyrA and parC genes from five ciprofloxacin-resistant strains of Haemophilus influenzae (MICs, 2 to 32 micrograms/ml) isolated from patients with cystic fibrosis and three ciprofloxacin-susceptible strains of H . influenzae (MICs, < or = 0.1 micrograms/ml) were determined . Four of the five resistant strains possessed at least one amino acid substitution in each of the GyrA and ParC fragments studied . The mutations identified in GyrA were a serine at residue 84 (Ser-84) to Leu or Tyr and Asp-88 to Asn or Tyr . ParC mutations were in positions exactly analogous to those identified in GyrA, namely, Ser-84 to Ile and Glu-88 to Lys . The Glu-88 to Lys ParC substitution was identified only in high-level ciprofloxacin-resistant strains . These mutations have been shown to be the origin of the observed resistance after transformation into ciprofloxacin-susceptible H . influenzae isolates . These results suggest that H . influenzae isolates require at least one amino acid substitution in both GyrA and ParC in order to attain significant levels of resistance to quinolones. Chemotherapy, 1996 Jul-Aug, 42(4), 240 - 7 Antibiotic susceptibility of clinical isolates of Streptococcus pneumoniae; Traub WH et al.; Ninety-three representative, recent clinical isolates of Streptococcus pneumoniae were examined for susceptibility to 9 antimicrobial drugs utilizing Mueller-Hinton agar (MHA) enriched with sheep blood and a hypercapnic atmosphere of incubation . One isolate was resistant to penicillin G (minimum inhibitory concentration, MIC = 2 micrograms/ml) and 6 isolates were of intermediate susceptibility to penicillin G (MICs = 0.125-0.25 microgram/ml) . The penicillin-G-resistant isolate was also resistant to cefuroxime (MIC = 4 micrograms/ml) and of intermediate susceptibility to cefotaxime (MIC = 1 microgram/ml) . This isolate was resistant to chloramphenicol (MIC = 16 micrograms/ml) as well . All 93 isolates were susceptible to teicoplanin and vancomycin . Two isolates each were resistant (MICs = 16 micrograms/ml) or moderately susceptible (MICs = 8 micrograms/ml) to chloramphenicol . Eight isolates were resistant to doxycycline (MICs > or = 8 micrograms/ml), whereas 2 isolates were of intermediate susceptibility to this antibiotic (MICs = 4 micrograms/ml) . Three isolates were resistant to erythromycin (MICs > or = 4 micrograms/ml), and 2 isolates showed reduced susceptibility to erythromycin (MICs = 2 micrograms/ml) . Chocolatized MHA antagonized the activity of teicoplanin and vancomycin against pneumococcal isolates . Haemophilus test and Wilkins-Chalgren media failed to support optimal growth of all pneumococcal isolates. Drugs, 1996 Jul, 52(1), 125 - 58 Cefuroxime axetil . A review of its antibacterial activity, pharmacokinetic properties and therapeutic efficacy; Perry CM et al.; Cefuroxime axetil is an oral cephalosporin which is rapidly hydrolysed to the active parent compound, cefuroxime . Cefuroxime has a broad spectrum of in vitro antibacterial activity which encompasses methicillin-sensitive staphylococci and the common respiratory pathogens Streptococcus pneumoniae, Haemophilus influenzae, Moraxella (Branhamella) catarrhalis and group A beta-haemolytic streptococci . Cefuroxime has broad spectrum activity against the beta-lactamase positive respiratory pathogens H . influenzae and M . catarrhalis; it is also active against penicillin-susceptible and -intermediate strains of S . pneumoniae . In clinical trials, cefuroxime axetil (administered twice daily) has been evaluated in the treatment of upper and lower respiratory tract infections and has demonstrated similar efficacy to established antibacterial agents, including amoxicillin/clavulanic acid and cefaclor . Five days' treatment with cefuroxime axetil was recently shown to be as effective as 10 days' treatment with either cefuroxime axetil or amoxicillin/clavulanic acid in patients with acute otitis media or acute bronchitis . Cefuroxime axetil was at least as effective as phenoxymethylpenicillin (penicillin V) in the treatment of patients with group A beta-haemolytic streptococcal tonsillopharyngitis . A number of studies have evaluated the efficacy of cefuroxime axetil as the oral component of intravenous to oral sequential therapy in hospitalised patients with lower respiratory tract infection . In each study patients received parenteral cefuroxime for approximately 2 days followed by cefuroxime axetil for 5 to 10 days . In comparative studies, cefuroxime sequential therapy was as effective as amoxicillin/ clavulanic acid sequential therapy and full courses of parenteral cefuroxime, cefotiam or cefoperazone . Adults with urinary tract infections and skin infections were also effectively treated with cefuroxime axetil, as were adults and adolescents with early stage lyme disease . Cefuroxime axetil is associated with a low incidence of adverse events, with gastrointestinal disturbances being the most frequently observed . Thus, cefuroxime axetil is an effective and convenient treatment for a wide range of infections and may be considered a therapeutic option when empirical treatment of community-acquired infections is required . Moreover, given the promising results of several intravenous/oral sequential treatment studies, cefuroxime axetil may also become established as an oral component of sequential treatment regimens. Endocrinology, 1996 Jul, 137(7), 2851 - 8 A disulfide bonding interaction role for cysteines in the extracellular domain of the thyrotropin-releasing hormone receptor; Cook JV et al.; The roles of disulfide and sulfhydryl groups in the specific binding of TRH to its receptor have been examined . In all TRH receptors (TRH-Rs) isolated from different species so far, there are only two extracellular cysteine residues (Cys98 in the extracellular loop between transmembrane helices 2 and 3 and Cys179 in the extracellular loop between transmembrane helices 4 and 5) that are in positions homologous to cysteine residues in other G protein-coupled receptors . Another Cys (Cys100) is located in close proximity to Cys98 at the interface between the first extracellular loop and third transmembrane domain . To assess the role of these TRH-R Cys residues in disulfide bonding interactions, they were mutated to either Ser or Ala . Six mutant receptors (Cys98Ser, Cys98Ala, Cys179Ser, Cys179Ala, Cys100Ser, and Cys100Ala) were expressed in COS-1 cells and tested for their ability to bind TRH and to activate total inositol phosphate (IP) formation . TRH-R mutants Cys100Ser and Cys100Ala showed TRH binding affinities and IP activation similar to the wild-type (WT) . In contrast, mutants Cys98Ser, Cys98Ala, Cys179Ser, and Cys179Ala showed no high affinity TRH binding . The potencies of Cys98Ala and Cysl79Ala as measured by IP stimulation were decreased by four orders of magnitude when compared with WT . Cys98Ser potency decreased by five orders of magnitude, whereas Cys179Ser showed no IP production . Northern blotting confirmed expression of all the mutant TRH-Rs at the messenger RNA (mRNA) level . An epitope tag derived from the Haemophilus influenza hemagglutinin protein was incorporated at the NH2 termini of the TRH-R WT and TRH-R Cys mutants to allow the independent assessment of cell surface expression of receptor protein . TRH-R mutants that failed to show receptor binding (Cys98Ser, Cys98Ala, Cys179Ala) showed WT levels of cell surface receptor expression, indicating that loss of receptor binding in these mutants is not attributable to loss of receptor expression . In contrast, cell surface expression of Cysl79Ser, which showed no ligand induced IP stimulation, could not be detected . Dithiothreitol, a disulfide bond reducing agent, and p-chloromercuribenzoic acid (p-CMB), a sulfhydryl blocking compound, reduced specific TRH binding in a dose-dependent manner . The inhibition of binding by dithiothreitol implies that the integrity of a disulfide bond is important for TRH binding to its receptor . The dramatic inhibition of TRH binding by p-CMB indicates that free sulfhydryl groups are also associated with the binding of the ligand to its receptor . This study presents evidence that a disulfide bond exists between Cys98 and Cys179 which is essential for maintaining the receptor in the correct conformation for ligand binding . Cys100 is not thought to have a disulfide bonding interaction role . Results obtained after chemical modification have shown that free sulfhydryl groups within the TRH-R may also have a role in ligand interactions. Am J Emerg Med, 1996 Jul, 14(4), 421 - 4 Epiglottitis in adults; Carey MJ; Acute epiglottitis has become a disease of adults, probably as a result of immunization of children against Haemophilus influenzae . This article is a review of the literature on epiglottitis, including signs and symptoms, investigation, differential diagnosis, and treatment in the emergency department . The microbiology is discussed and the importance of prophylaxis in exposed persons is stressed. J Pediatr, 1996 Jul, 129(1), 72 - 80 Early-onset sepsis in very low birth weight neonates: a report from the National Institute of Child Health and Human Development Neonatal Research Network; Stoll BJ et al.; OBJECTIVE: Early-onset sepsis (occurring within 72 hours of birth) is included in the differential diagnosis of most very low birth weight (VLBW) neonates . To determine the current incidence of early-onset sepsis, risk factors for disease, and the impact of early-onset sepsis on subsequent hospital course, we studied a cohort of 7861 VLBW neonates (401 to 1500 gm) admitted to the 12 National Institute of Child Health and Human Development (NICHD) Neonatal Research Network centers during a 32-month period (1991-1993) . METHODS: The NICHD Neonatal Research Network maintains a prospectively collected registry on all VLBW neonates born or cared for at participating centers . Data from this registry were analyzed retrospectively . RESULTS: Blood culture-proven early-onset sepsis was uncommon, occurring in only 1.9% of VLBW neonates . Group B streptococcus was the most frequent pathogen associated with early-onset sepsis (31%), followed by Escherichia coli (16%) and Haemophilus influenzae (12%) . Decreasing gestational age was associated with increased rates of infection . Antibiotic therapy for suspected sepsis is frequently initiated at birth in VLBW neonates . Almost half of the infants in this cohort were considered to have clinical sepsis and continued to receive antibiotics for 5 or more days, despite a negative blood culture result in 98% of cases . These findings underscore the difficulty of ruling out sepsis in the symptomatic immature neonate and the special concern for culture-negative clinical sepsis in the face of maternal antibiotic use . Neonates with early-onset sepsis were significantly more likely to have subsequent comorbidities, including severe intraventricular hemorrhage, patent ductus arteriosus, and prolonged assisted ventilation . Although 26% of VLBW neonates with early-onset sepsis died, only 4% of the 950 deaths that occurred in the first 72 hours of life were attributed to infection . For those infants discharged alive, early-onset sepsis was associated with a significantly prolonged hospital stay (86 vs 69 days; p <0.02) . CONCLUSIONS: Early-onset sepsis remains an important but uncommon problem among VLBW preterm infants . Improved diagnostic strategies are needed to enable the clinician to distinguish between the infected and the uninfected VLBW neonate with symptoms and to target continued antibiotic therapy to those who are truly infected. Infect Immun, 1996 Jul, 64(7), 2627 - 34 Cloning and characterization of the catalase gene of Neisseria gonorrhoeae: use of the gonococcus as a host organism for recombinant DNA; Johnson SR et al.; The structural gene for the catalase of Neisseria gonorrhoeae was cloned into a Kat- strain of that organism by using a recombinant vector derived from one of the beta-lactamase-specifying plasmids found in that organism . The kat gene was then successfully subcloned into both pUC8 and pGB2, transformed into Escherichia coli, and shown to complement the E . coli katE mutants UM2 and UMRl . The gene was subsequently mutagenized and returned to the gonococcus to generate a Kat- strain that was phenotypically identical to the strain originally used to clone the gene . The sequence of the gene and the derived amino acid sequence showed that the gonococcal kat gene closely resembles the hktE gene of Haemophilus influenzae . The sequence of the promoter region of the gonococcal kat gene is unusual and may explain the extremely high, loosely regulated expression of the gene. Br J Obstet Gynaecol, 1996 Jul, 103(7), 664 - 9 Bacterial 16S rDNA polymerase chain reaction in the detection of intra-amniotic infection; Jalava J et al.; OBJECTIVE: Bacterial polymerase chain reaction (PCR) was used to detect early subclinical intraamhiotic infection . We used universal primers which amplify a DNA fragment of 16S ribosomal DNA (rDNA) from all known bacteria and sequenced the positive samples to identify the bacterial species . DESIGN: Transabdominally obtained amniotic fluid samples from 20 pregnant women with prelabour rupture of the fetal membranes (PROM), showing no signs of clinical infection, and 16 control samples were analysed with universal bacterial PCR . In addition, routine bacterial culture and amniotic fluid glucose were studied . RESULTS: Out of 20 PROM patients, five were positive in the PCR . PCR detected Ureaplasma urealyticum in two cases, Haemophilus influenzae in one case, Streptococcus oralis in one case and Fusobacterium sp . in one case . Only two of these were positive in a routine bacterial culture . Both were multibacterial infections, which caused discrepancies between the PCR and culture results . Two patients developed infectious complications: both were identified with the PCR assay . Amniotic fluid glucose was lower in PCR positive patients compared with PCR negative patients . CONCLUSION: Bacterial 16S rDNA PCR, in properly controlled conditions, promises to be a fast and reliable test for early intra-amniotic infection especially concerning Ureaplasma urealyticum. Lett Appl Microbiol, 1996 Jul, 23(1), 47 - 8 Anomalous but helpful findings from the BBL Crystal ID kit with Haemophilus spp; Hamilton-Miller JM et al.; Fourteen strains of Haemophilus (12 H . influenzae, 1 H . parainfluenzae and 1 H . aphrophilus) were processed in BBL Crystal ID Enteric/Nonfermenter, API 20E and API 20NE kits, to determine whether the BBL kit misidentifies, as API kits may do, Haemophilus spp . as Pasteurella spp . The 13 H . influenzae and H . parainfluenzae strains produced uninterpretable colour reactions in the Crystal kit, thus signalling that an inappropriate species had been tested . On the other hand, the API kits (especially 20NE) often confidently "identified' Haemophilus spp . as Pasteurella spp., giving no warning that this was a misidentification. J Biol Chem, 1996 Jun 28, 271(26), 15373 - 80 Purification, cloning, and expression of a cytidine 5'-monophosphate N-acetylneuraminic acid synthetase from Haemophilus ducreyi; Tullius MV et al.; An N-acetylneuraminic acid cytidylyltransferase (EC 2.7.7.43) (CMP-NeuAc synthetase) was isolated from a Haemophilus ducreyi strain 35000 cell lysate and partially characterized . The enzyme catalyzes the reaction of CTP and NeuAc to form CMP-NeuAc, which is the nucleotide sugar donor used by sialyltransferases . Previous studies have shown that the outer membrane lipooligosaccharides of H . ducreyi contain terminal sialic acid attached to N-acetyllactosamine and that this modification is likely important to its pathogenesis . Therefore, to investigate the role of sialic acid in H . ducreyi pathogenesis, the gene encoding the CMP-NeuAc synthetase was cloned using degenerate oligonucleotide probes derived from NH2-terminal sequence data, and the nucleotide sequence was determined . The derived amino acid sequence of the CMP-NeuAc synthetase gene has homology to other CMP-NeuAc synthetases and to a lesser extent to CMP-2-keto-3-deoxy-D-manno-octulosonic acid synthetases . The gene was cloned into a T7 expression vector, the protein expressed in Escherichia coli, and purified to apparent homogeneity by anion exchange, Green 19 dye, and hydrophobic interaction chromatography . The final step yielded 20 mg of pure protein/liter of culture . The protein has a predicted molecular mass of 25440.6 Da, which was confirmed by electrospray mass spectrometry (Mexpt = 25439.9 +/- 1.4 Da) . The enzyme appears to exist as a dimer by size exclusion chromatography . In contrast to other bacterial CMP-NeuAc synthetases, the H . ducreyi enzyme exhibited a different substrate specificity, being capable of also using N-glycolylneuraminic acid as a substrate. MMWR Morb Mortal Wkly Rep, 1996 Jun 21, 45(24), 508 - 13 National, state and urban area vaccination coverage levels among children aged 19-35 months--United States, July 1994-June 1995; The complete nucleotide sequence of bacteriophage HP1 DNA; Department of Biochemistry, The Johns Hopkins University School of Hygiene and Public Health, Baltimore, MD 21205, USAThe complete nucleotide sequence of the temperate phage HP1 of Haemophilus influenzae was determined . The phage contains a linear, double-stranded genome of 32 355 nt with cohesive termini . Statistical methods were used to identify 41 probable protein coding segments organized into five plausible transcriptional units . Regions encoding proteins involved in recombination, replication, transcriptional control, host cell lysis and phage production were identified . The sizes of proteins in the mature HP1 particle were determined to assist in identifying genes for structural proteins . Similarities between HP1 coding sequences and those in databases, as well as similar gene organizations and control mechanisms, suggest that HP1 is a member of the P2-like phage family, with strong similarities to coliphages P2 and 186 and some similarity to the retronphage Ec67. J Immunol Methods, 1996 Jun 14, 193(1), 1 - 7 Simple determination of polysaccharide specific antibodies by means of chemically modified ELISA plates; Zielen S et al.; A new ELISA technique using Nunc CovaLink NH microtiter plates has been developed to measure anticapsular polysaccharide specific antibodies . Capsular polysaccharide (PS) of Haemophilus influenzae type b (PRP) and pneumococcal antigens types 3, 6, 8, 14, 19, 23 were immobilized on CovaLink NH . These are modified plates with secondary amino groups bound to their surface which, in the presence of a water-soluble carbodiimide as coupling reagent, facilitate the direct binding of polysaccharides . We compared the binding characteristics of PS antigens to CovaLink NH and a conventional polystyrene ELISA plate . Checkerboard titration of PS antigens between 0.04-30 micrograms/ml clearly demonstrated that with Covalink NH optimal binding of a pooled serum from immunized donors was achieved for all PS antigens tested at a concentration of 1 microgram/ml, while binding of PS to the conventional plate was rather poor even at concentrations of 30 micrograms/ml . The CVs for the ELISA ranged from 1.1 to 2.8% for intra-assay comparisons and from 3.6 to 7.3% for inter-assay comparisons . In addition, when PRP-IgG antibodies were determined with the CovaLink NH ELISA and compared with the Farr assay an acceptable correlation ( r = 0.89, p < 0.0001) was obtained . The technique described provides a simple and sensitive tool for evaluating specific immunity to PS antigens. Gene, 1996 Jun 12, 172(1), 71 - 3 Determination of the cos sequence of the mature genome of S2/HP1 type B bacteriophage of Haemophilus influenzae; Skowronek K et al.; The cos region of Haemophilus influenzae phage HP1/S2 type B has been cloned and its nucleotide (nt) sequence determined . The nt sequence of the cohesive ends (cos) and whole cos site of type-B phage have been compared to corresponding sequences of the HP1c1 phage . The results of a search for symmetry elements and IHF-binding sites in this region are presented. Minerva Pediatr, 1996 Jun, 48(6), 235 - 44 {Haemophilus influenzae disease in childhood . Comment about case reports of meningitis}; Ranno O et al.; The authors describe a series of Haemophilus influenzae meningitis in childhood, obtained with a retrospective analysis of the cases of bacterial meningitis admitted to Isolamento Pediatrico department of "A . Gemelli" Polyclinic in Rome, from January 1, 1970 to December 31, 1994 . Haemophilus influenzae resulted the second agent in frequency (first was Neisseria meningitidis) . Main features were: no patient was older than 5 years, and most of them were less than 2 years old; clinical feature was aspecific in the first year of life, it was typical of bacterial meningitis in older children; blood culture and detection of bacterial antigens in cerebrospinal fluid (CSF) were useful for etiological diagnosis, supporting CSF culture; clinical course was characterized by many complications, but no case was lethal and incidence of sequelae at discharge was low; C reactive protein was effective as index of inflammation and as indicator of arising complications; chosen antibiotics were efficacious, but frequency of antibiotic resistance, especially to beta-lactams, was found to be increasing; results of dexamethasone therapy were not of univocal interpretation . The authors are in favour of spreading of vaccination against Haemophilus influenzae in Italy, too, in order to eradicate this disease, as experiences in other countries are successful, and of setting up of the combined vaccines, in order to increase parents' compliance to vaccinal practices. Tohoku J Exp Med, 1996 Jun, 179(2), 111 - 21 The incidence of respiratory tract pathogens and antimicrobial susceptibilities of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella (Branhamella) catarrhalis isolated between 1990 and 1993; Nishioka K et al.; Using a quantitative culture of sputum, the incidence of pathogenic bacteria in respiratory infection in our laboratory between 1990 and 1993 were investigated . While Haemophilus influenzae, Streptococcus pneumoniae and Moraxella (Branhamella) catarrhalis were isolated at high rates (67-78%) from the specimens of outpatients throughout the study period, the incidence of S pneumoniae has increased gradually . The antimicrobial susceptibilities of these three pathogens were examined with the agar dilution method . A marked increase of penicillin (PC) resistant S . pneumoniae (MIC > or = 0.1 microgram/ml) was observed with a resistance rate of 2.1% in 1990 and 25% in 1993 . Resistance to erythromycin (EM, MIC > or = 1.56 micrograms/ml) was 8.5% in 1990 but then increased to 34% in 1992 . Most of the PC resistant isolates were resistant to multidrugs such as EM, minocycline and clindamycin . The MICs of all beta-lactams examined for S . pneumoniae increased along with the MICs of PC, though the level varied between drugs . The rates of beta-lactamase positive H . influenzae gradually decreased, being 14.3% in 1990 and 7.4% in 1993, whereas those of M . (B) catarrhalis were consistently high (> 90%) every year . In addition to beta-lactamase production, the emergence of strains of H . influenzae and M . (B) catarrhalis resistant to new quinolone drugs should be noted. Vaccine, 1996 Jun, 14(9), 905 - 9 The use of Haemophilus influenzae type b-tetanus toxoid conjugate vaccine mixed with diphtheria-tetanus-pertussis vaccine in Gambian infants; Mulholland EK et al.; In preparation for an efficacy trial of PRP-T Haemophilus influenzae type b conjugate vaccine, 251 Gambian infants were randomized to receive three doses of PRP-T and diphtheria-tetanus-pertussis (DTP) vaccines at 2, 3 and 4 months of age, either by separate injections, or combined in the same syringe . One month after the third dose, there was no difference between anti-PRP levels in those infants who received the vaccines separately (GMT 5.83 micrograms ml-1), and those who received the vaccines combined (GMT 5.57 micrograms ml-1) . The proportions achieving levels of 1.0 microgram ml-1 were 89% and 92% in the "separate" and "combined" vaccine groups, respectively . There were no significant differences between groups in levels of antibody to diphtheria or tetanus . Geometric mean titres of antibody directed against pertussis antigens in the "separate" and "combined" groups were as follows: pertussis toxin 14.2 and 13.1 ELISA units (EU) ml-1; filamentous haemagglutinin 12.2 and 9.7 EU ml-1; pertactin 17.2 and 9.0 EU ml-1 (P < 0.05), fimbrial 2/3 antigens 449 and 364 EU ml-1 . The combination of PRP-T and DTP in the syringe prior to administration is safe and immunogenic . The lower levels of anti-pertussis antibody are of unknown clinical significance. Eur J Clin Microbiol Infect Dis, 1996 Jun, 15(6), 472 - 7 Modified latex agglutination test for rapid detection of Streptococcus pneumoniae and haemophilus influenzae in cerebrospinal fluid and direct serotyping of Streptococcus pneumoniae; Singhal A et al.; A modified latex agglutination test was designed and evaluated for the rapid detection of Streptococcus pneumoniae and haemophilus influenzae type b capsular antigens, and for direct serotyping of Streptococcus pneumoniae in the cerebrospinal fluid . Reagents were prepared by sensitizing latex particles with Omniserum (against 83 capsular serotypes of pneumococci) and Haemophilus influenzae type b burro antiserum . For serotyping reagents, latex particles were similarly coated with nine pneumococcal pool (a to I) antisera and 46 individual pneumococcal serogroup/serotype specific antisera . The test was performed on cerebrospinal fluid from 298 patients with suspected meningitis . Serotyping was done directly on untreated cerebrospinal fluid samples showing positive reactions with the Omniserum reagent . Pneumococcal or Haemophilus influenzae type b antigens were detected in 41 patients; in 32 of these the etiology was established by culture and in 2 by smear examination . Five of the remaining seven cases were judged clinically and by cytological examination of cerebrospinal fluid to have partially treated bacterial meningitis . In two cases the test was false positive . The overall sensitivity and specificity of the latex agglutination test for the detection of Streptococcus pneumoniae and Haemophilus influenzae type b antigens was 100% and 96.8% respectively . The commonest pneumococcal serotypes were type 1 (30%), types 6 and 19 (10% each) . The latex agglutination test is rapid and simple to perform, yielding serotype data directly by testing of cerebrospinal fluid. Zentralbl Bakteriol, 1996 Jun, 284(1), 47 - 51 A comparative study of the major outer membrane proteins of the avian haemophili and Pasteurella gallinarum; Hartmann L et al.; A polyclonal antibody prepared against the 35 kDa outer membrane protein (a putative porin) of Pasteurella (P.) multocida revealed binding to the 36 kDa major outer membrane protein (major Omp) of Haemophilus (H.) paragallinarum, to the 38 kDa major Omp of P.gallinarum, to the 39 kDa major Omp of P.volantium and to the 38.5 kDa major Omp of P . avium in immunoblotting studies . Comparison of N-terminal amino acid sequences also confirmed the relationship between the major Omps of most of the members of the family Pasteurellaceae. J Antimicrob Chemother, 1996 Jun, 37 Suppl C, 115 - 24 Efficacy, safety and tolerability of azithromycin versus roxithromycin in the treatment of acute lower respiratory tract infections; Laurent K; In an open, multicentre study, the clinical and bacteriological efficacy, safety and tolerance of azithromycin and roxithromycin were compared in a total of 204 adults with acute lower respiratory tract infections (LRTIs) {acute bronchitis, acute infectious exacerbations of chronic bronchitis (AIECBs), or pneumonia} . Following treatment with 500 mg/day azithromycin administered orally once daily for 3 days, a satisfactory clinical response of cure or improvement was recorded in 91/99 (91.9%) evaluable patients at the post-therapy evaluation (day 10-14) . Of the 94 evaluable patients treated with roxithromycin (150 mg given orally twice daily for 10 days), 82 (87.2%) were classified as cured or improved at post-therapy . The main pathogens isolated before treatment were Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus species, Haemophilus influenzae and Moraxella catarrhalis . In the 46 azithromycin-treated patients evaluated both clinically and bacteriologically, 92.0% of pathogens were eradicated; H . influenzae persisted in one azithromycin-treated patient with acute bronchitis who was classed as clinically improved . In the roxithromycin group, 81.1% of the pathogens were eradicated in 35 patients; S . aureus persisted in one clinically cured patient with acute bronchitis, and H . influenzae persisted in one patient with AIECB and one with pneumonia, and Haemophilus species in one with AIECB, who were all classified as clinically improved . Azithromycin was well tolerated with a lower incidence of adverse events than that recorded in the roxithromycin treatment group . Treatment was not discontinued due to adverse events in any of the azithromycin-treated patients, whereas two roxithromycin-treated patients were withdrawn from treatment due to vomiting and/or dyspepsia. J Antimicrob Chemother, 1996 Jun, 37 Suppl C, 103 - 13 A randomized, comparative study to evaluate the efficacy and tolerability of a 3-day course of azithromycin versus a 10-day course of co-amoxiclav as treatment of adult patients with lower respiratory tract infections; Zachariah J; Clinical and bacteriological efficacy and tolerability of azithromycin (500 mg once daily for 3 days) and those of a 10-day regimen of co-amoxiclav (37 mg three times daily) were evaluated in a large-scale, double-blind comparative study of 369 patients (> or = 18 years old) with acute lower respiratory tract infections . After treatment, 165/173 (95%) azithromycin- and 166/173 (96%) co-amoxiclav-treated patients had responded satisfactorily (cure or improvement) . Baseline pathogens (mainly Streptococcus pneumoniae and Haemophilus influenzae) were eradicated in 82/82 (100%) azithromycin- and 73/74 (99%) co-amoxiclav-treated patients who were bacteriologically assessable . Adverse events, which were predominantly of mild to moderate severity and mostly affected the gastrointestinal system, were recorded in 13/186 (7%) azithromycin- and 19/183 (10%) co-amoxiclav-treated patients . Only two (1%) azithromycin-treated patients discontinued treatment due to adverse events compared with eight (4%) who received co-amoxiclav . The results show that azithromycin at a dose of 500 mg once daily for 3 days is an effective and safe alternative to a 10-day, three-times-daily course of co-amoxiclav in the treatment of lower respiratory tract infections in adults. J Antimicrob Chemother, 1996 Jun, 37 Suppl C, 93 - 101 Once-daily, 3-day azithromycin versus a three-times-daily, 10-day course of co-amoxiclav in the treatment of adults with lower respiratory tract infections: results of a randomized, double-blind comparative study; Gris P; A 3-day regimen of azithromycin (500 mg once daily) and a 10-day regimen of co-amoxiclav (625 mg three times daily) were compared in a double-blind study of 67 patients with acute infectious exacerbations of chronic bronchitis (AIECBs, n = 54), acute bronchitis (n = 7), or pneumonia (n = 6) . In patients treated with azithromycin, satisfactory clinical responses (cure or improvement) were seen in 24/28 (86%) patients with AIECBs, 2/4 (50%) with acute bronchitis and 2/2 (100%) with pneumonia . Responses were satisfactory in 24/26 (92%), 4/4 (100%) and 4/4 (100%) patients, respectively, receiving co-amoxiclav . Streptococcus pneumoniae and Haemophilus influenzae were the commonest pathogens isolated at baseline . At the end of treatment, baseline pathogens were eradicated in 9/10 microbiologically-assessable patients treated with azithromycin and in 10/10 treated with co-amoxiclav . Adverse events related or possibly related to treatment occurred in five patients in each treatment group; the majority of these events affected the gastrointestinal system . One patient in each treatment group discontinued therapy because of adverse events . The study, therefore, demonstrates that 500 mg azithromycin administered once daily for 3 days is as efficacious and well tolerated as co-amoxiclav given three times daily for 10 days in the domiciliary treatment of adults with acute lower respiratory tract infections. J Antimicrob Chemother, 1996 Jun, 37 Suppl C, 9 - 19 Correlation of increased azithromycin concentrations with phagocyte infiltration into sites of localized infection; Girard AE et al.; Azithromycin reaches high concentrations in phagocytic and other host cells, suggesting that they may transport this agent to specific sites of infection . Models of localized infection (Haemophilus influenzae middle ear infection in gerbils, Streptococcus pyogenes implanted contaminated paper disc and Streptococcus pneumoniae pneumonia in mice) that induced severe inflammatory response after challenge were used to explore this hypothesis . Animals were given a single 100 or 50 mg/kg po dose of azithromycin at various times from 2 to 120 h following introduction of a pathogen or sterile medium . When azithromycin was given during a period of little or no inflammation, there was marginal difference between concentrations found in infected or non-infected sites (bulla, disc, lung) . However, when the compound was given during a period of inflammation, considerably higher drug concentrations were found in infected sites than in non-infected sites at 5-24 h after dosing (0.38-0.44 mg/c compared with 0.07-0.14 mg/L of bulla wash; 1.01-1.75 micrograms compared with < or = 0.01-0.03 microgram at the disc site; 1.72-5.28 mg/kg compared with 0.7-1.53 mg/kg of lung) . When the observation periods were extended to include 48, 56 or 96 h after dosing, the ratio of azithromycin infection site concentration: serum concentration steadily increased with time in all model systems (middle ear, implanted disc and pneumonia), reflecting the maintenance of concentrations at the sites of infection, while serum concentrations declined . Bioassay of cell pellets and supernatants, obtained from pooled bulla washes of gerbils treated with azithromycin during a period of inflammation, revealed that cellular components accounted for about 75% of the azithromycin detected . These data show that increased azithromycin concentrations occur at sites of localized infection . This correlates with the presence of inflammation and is associated with the cellular components of the inflammatory response . Therefore, phagocytes may be important vehicles for delivering azithromycin to and sustaining azithromycin concentrations at sites of infection. J Antimicrob Chemother, 1996 Jun, 37 Suppl C, 1 - 8 Azithromycin--review of key chemical, pharmacokinetic and microbiological features; Lode H et al.; One of the chemical features that distinguishes the 15-membered ring azalide azithromycin from the 14-membered ring macrolide compound erythromycin is the former's increased stability at acid pH . Azithromycin also differs pharmacokinetically from erythromycin, an important feature being azithromycin's ability to achieve high tissue concentrations, with the agent being delivered to the sites of infection by direct uptake and by targeted delivery via phagocytes . High tissue concentrations are maintained for prolonged periods because of azithromycin's long half-life, leading to once-daily dosing for 3 or 5 days . Notable microbiological features of azithromycin are in-vitro activity against many pyogenic bacteria (e.g . Neisseria gonorrhoeae and Moraxella catarrhalis), as well as organisms against which beta-lactam antibiotics are usually ineffective (e.g . Legionella and Chlamydia spp.), organisms that are resistant to benzylpenicillin and erythromycin (e.g . Haemophilus influenzae) and organisms for which satisfactory therapy is limited (e.g . Toxoplasma gondii and the Mycobacterium avium-intracellulare complex) . These properties of azithromycin suggest that it might be a useful agent for the treatment of a wide range of bacterial infections. Vaccine, 1996 Jun, 14(8), 811 - 6 Safety and immunogenicity of a recombinant hepatitis B vaccine administered to infants at 2, 4 and 6 months of age . The Kaiser-UCLA Vaccine Study Group; Greenberg DP et al.; A recombinant hepatitis B vaccine was administered to over 5000 infants in a prospective, randomized and blinded study . Infants were given either recombinant hepatitis B vaccine (Engerix-B, SmithKline Beecham Pharmaceuticals, 10 micrograms dose-1) or a Haemophilus influenzae type b (Hib) conjugate vaccine at 2, 4 and 6 months of age simultaneously with diphtheria-tetanus-pertussis and oral polio vaccines . Adverse reactions were ascertained by parental reports and interviews, and review of medical records . Blood specimens collected from 269 infants given hepatitis B vaccine were assayed for antibody to hepatitis B surface antigen (anti-HBs) by enzyme immunoassay . Infants given hepatitis B vaccine experienced low rates of adverse reactions that were similar or lower than the rates in infants given Hib conjugate vaccine . The geometric mean anti-HBs concentrations were 9.6 mIU ml-1 after one dose, 333 mIU ml-1 after two doses and 1812 mIU ml-1 after three doses (99% had levels > or = 10 mIU ml-1) . Antibody responses to diphtheria and tetanus toxoids were unaffected by simultaneous administration of hepatitis B or Hib conjugate vaccine . Engerix-B vaccine was safe and immunogenic when given with other routine childhood immunizations at 2, 4 and 6 months of age, and should provide long-term protection against hepatitis B virus infection. J Vet Med Sci, 1996 Jun, 58(6), 559 - 61 Pathogenicity of Haemophilus parasuis serovars 4 and 5 in contact-exposed pigs; Amano H et al.; The pathogenicities of Haemophilus parasuis strains SW124 (serovar 4) and Nagasaki (serovar 5) were examined by contact-exposure of specific pathogen-free (SPF) pigs . Ten pigs were divided into three groups . Two of four pigs in the first group were inoculated intranasally (IN) with 2 x 10(8) CFU of strain SW124, and the other two pigs were mingled with these IN-exposed ones . All the four pigs were subclinically infected in this group . The four pigs of the second group were likewise exposed to strain Nagasaki (two IN-inoculated pigs with 3 x 10(8) CFU of strain Nagasaki) . All four pigs in this group died of Glasser's disease . Two pigs kept as controls showed neither abnormality nor positive H . parasuis isolation. Mol Microbiol, 1996 Jun, 20(6), 1273 - 86 A novel DnaJ-like protein in Escherichia coli inserts into the cytoplasmic membrane with a type III topology; Clarke DJ et al.; We describe a novel Escherichia coli protein, DjlA, containing a highly conserved J-region motif, which is present in the DnaJ protein chaperone family and required for interaction with DnaK . Remarkably, DjlA is shown to be a membrane protein, localized to the inner membrane with the unusual Type III topology (N-out, C-in) . Thus, DjlA appears to present an extremely short N-terminus to the periplasm and has a single transmembrane domain (TMD) and a large cytoplasmic domain containing the C-terminal J-region . Analysis of the TMD of DjlA and recently identified homologues in Coxiella burnetti and Haemophilus influenzae revealed a striking pattern of conserved glycines (or rarely alanine), with a four-residue spacing . This motif, predicted to form a spiral groove in the TMD, is more marked than a repeating glycine motif, implicated in the dimerization of TMDs of some eukaryotic proteins . This feature of DjlA could represent a promiscuous docking mechanism for interaction with a variety of membrane proteins . DjlA null mutants can be isolated but these appear rapidly to accumulate suppressors to correct envelope and growth defects . Moderate (10-fold) overproduction of DjlA suppresses a mutation in FtsZ but markedly perturbs cell division and cell-envelope growth in minimal medium . We propose that DjlA plays a role in the correct assembly, activity and/or maintenance of a number of membrane proteins, including two-component signal-transduction systems. Clin Exp Allergy, 1996 Jun, 26(6), 665 - 76 Bronchial inflammation and the common cold: a comparison of atopic and non-atopic individuals; Trigg CJ et al.; BACKGROUND: Cold virus infections are associated with asthma attacks and with increased bronchial responsiveness even in normal subjects . Possible mechanisms include epithelial damage, interaction with adhesion molecules or with T-helper cell subsets . OBJECTIVE: To determine whether colds increase lower airway inflammation, comparing atopic with non-atopic normal subjects . METHODS: Thirty healthy volunteers (15 atopic) took part . Baseline tests included viral serology, microbiological culture and polymerase chain reaction for rhinovirus infection (HRV-PCR), histamine bronchial provocation and bronchoscopy . Twenty subjects (eight atopic) underwent repeat tests when they developed a cold . RESULTS: Forced expiratory volume in one second (FEV1) was significantly lower during colds (-0.19 L {95% confidence interval -0.10, -0.29}, P = 0.0004) and there was a significant increase in bronchial responsiveness (+0.62 doublings of the dose-response slope {+0.24, +1.00}, P = 0.003) . Eight subjects (two atopic) had a diagnosed viral infection: two HRV, three coronavirus (HCV), one HRV + HCV, one parainfluenza III (PI) and one respiratory syncytial virus (RSV) (also Haemophilus influenzae) . In biopsies, during colds, total eosinophils (EGI+) increased significantly (geometric mean 6.73-fold {1.12,40.46}, P = 0.04) . Activated eosinophils (EG2+) only increased significantly in the subgroup without diagnosed viral infection and particularly in atopic rhinitics . T-suppressor (CD8+) cells also increased significantly (median + 178.3 cells mm2, P = 0.004) . Epithelial expression of intercellular adhesion molecule-1 (ICAM-1) expression increased in four atopic rhinitics during colds . Bronchial washings showed a significant increase in neutrophils (GM 1.53-fold {1.04,2.25}, P = 0.02) . CONCLUSION: Lower airway inflammation was present in atopic and non-atopic normal subjects with colds . Atopic subjects differed in that they were less likely to have positive virological tests and were more likely to show activated eosinophilia in the lower airway, despite a similar spectrum of symptoms. J Chemother, 1996 Jun, 8(3), 193 - 9 In vitro activity of cefdinir against respiratory pathogens isolated in Sicily with reference to beta-lactamase production; Blandino G et al.; The in vitro activity of cefdinir (CI-983, FK-482), an orally absorbed aminothiazolyl cephalosporin, was evaluated against all 287 strains of Haemophilus influenzae, Haemophilus parainfluenzae, Moraxella catarrhalis, Streptococcus pneumoniae and Streptococcus pyogenes in comparison with cefaclor, cefuroxime, amoxicillin, amoxicillin-clavulanic acid, erythromycin and cotrimoxazole . The bactericidal activity of cefdinir, cotrimoxazole, amoxicillin-clavulanic acid and erythromycin was determined against H . influenzae, M . catarrhalis and S . pneumoniae . With the exception of one beta-lactamase negative ampicillin-resistant strain of H . influenzae (resistant to all antibiotics tested), no resistance to cefdinir was observed (MIC < or = 1 mg/l) . Cefdinir was active against H . influenzae, H . parainfluenzae and M . catarrhalis regardless of whether or not they produced beta-lactamase . In general, the inhibitory concentrations of cefdinir against H . influenzae, H . parainfluenzae and M . catarrhalis were similar to those of amoxicillin/clavulanic acid, one or two dilutions lower than those of cefuroxime and four dilutions lower than those of cefaclor and cotrimoxazole . Against S . pneumoniae and S . pyogenes cefdinir had the same activity as cefuroxime and amoxicillin but was more effective than the other antibiotics tested . Kinetic studies showed that cefdinir was rapidly bactericidal at concentrations 2 and 4 times the minimum inhibitory concentration (MIC): a reduction of 99.9% in CFU values was generally observed after 6-8 h. J Chemother, 1996 Jun, 8(3), 178 - 87 Comparison of the serum-supplemented Todd-Hewitt and the new Haemophilus test media for broth microdilution susceptibility testing of Streptococcus pneumoniae; Roger M et al.; Horse serum-supplemented Todd-Hewitt broth (STH) in use at Hopital Ste-Justine for the last 12 years was compared to the recently proposed Haemophilus test medium (HTM), for broth microdilution susceptibility testing of Streptococcus pneumoniae . One hundred and twenty S . pneumoniae isolates from pediatric clinical specimens were used in this study . In general, the minimum inhibitory concentrations (MICs) in STH for 15 antimicrobial agents were quite comparable to those determined in HTM but tended to be higher . Drugs which generated MICs within +/- 1 log2 concentration differences in both media included penicillin, ampicillin, oxacillin, cefuroxime, cefotaxime, cefixime, clindamycin, chloramphenicol, trimethoprim-sulfamethoxazole, rifampin, ciprofloxacin and vancomycin . Cefaclor and tetracycline MICs tended to be > or = 2 log2 dilutions higher with STH for most of the isolates tested, while erythromycin MICs were often 2 log2 dilutions lower with STH than with HTM . Despite some differences in MICs noted above, few very major (0.4%), major (0.2%) and minor interpretive category errors (4.4%) were observed . The visual reading of the MICs for most of the 120 clinical isolates tested was generally easier in STH which was superior in supporting best the bacterial growth as detected by spectrophotometry . The risk of false susceptibility is thus decreased by using STH rather than HTM; furthermore, STH is free of the technical problems of the lysed horse blood Mueller-Hinton (LHB-MH) recommended by the NCCLS. J Chemother, 1996 Jun, 8(3), 171 - 7 Brodimoprim: effects of subminimal inhibitory concentrations on virulence traits of respiratory and urinary tract pathogens, and on plasmid transfer and stability; Marchese A et al.; The effects of brodimoprim, a new trimethoprim analogue, on several virulence traits of respiratory and urinary tract pathogens exposed to sub-lethal levels of the drug was studied . Adherence to tracheal epithelial cells was inhibited by brodimoprim in Klebsiella pneumoniae (41-67% reduction), Moraxella catarrhalis (87-90%) and Haemophilus influenzae (0-53%), while in Streptococcus pneumoniae binding was unaffected . With buccal epithelial cells the comparison between treated and control bacteria indicated statistically significant reduction in adherence with both S.pneumoniae and H.influenzae, (P < 0.015) . With M.catarrhalis and Streptococcus pyogenes only marginal changes were detected (P > 0.05) . Exoenzyme and capsule production were assessed in at least three isolates of diverse respiratory pathogens grown in the presence of sub-lethal levels of the new agent . The drug affected protease and beta-hemolysin (alpha-toxin) production in both oxacillin-susceptible and -resistant S.aureus . On the contrary, synthesis of lipase, DNase, coagulase, and beta-lactamase (S.aureus), pneumolysin (S.pneumoniae), streptolysin S, DNase, and protease (S.pyogenes), capsule (K.pneumoniae, H.influenzae and S.pneumoniae), and beta-lactamase (K.pneumoniae, H.influenzae and M.catarrhalis) were not inhibited by subminimal inhibitory concentrations (sub-MICs) of the drug . Finally, motility was blocked in urinary pathogens E.coli, P.mirabilis and P.aeruginosa, while in this latter microorganism pigment production was also affected . High molecular weight low-copy F'lac, and low molecular weight high-copy pHSG298 plasmids were eliminated from E.coli treated with sub-MIC concentrations of brodimoprim . The incidence and cured cells ranged from 9% for F'lac to 23% for pHSG298 . F'lac transfer was also inhibited by the drug . When conjugation was carried out with bacteria exposed to brodimoprim (5XMIC), a reduction (50%) in the number of recombinants was noted in comparison to the control . The fact that brodimoprim interferes with the expression of some virulence traits, in particular with adherence, at sub-MIC levels may assist the drug in eradicating respiratory pathogens from the epithelial lining, thus diminishing the probability of reinfection. Bone Marrow Transplant, 1996 Jun, 17(6), 1149 - 55 Haemophilus influenzae type b (HIB)-conjugate immunization before bone marrow harvest in autologous bone marrow transplantation; Molrine DC et al.; Immune reconstitution following autologous bone marrow transplantation (ABMT) is characterized by defects in B cell and T cell function and loss of specific antibody . In the late post-transplant period, patients are at risk for infections with polysaccharide encapsulated organisms and respond poorly to polysaccharide vaccines . We examined whether immunizing ABMT patients before bone marrow (BM) harvest enhanced the early recovery of specific antibody . Twelve patients were immunized before BM harvest with Haemophilus influenzae type b (HIB)-conjugate, tetanus toxoid and polysaccharide pneumococcal vaccines . Forty-one comparable ABMT patients not immunized prior to BM harvest were also studied . Following ABMT, both groups of patients were immunized with HIB-conjugate and tetanus toxoid vaccines at 3, 6, 12 and 24 months and with pneumococcal vaccine at 12 and 24 months . Patients immunized before BM harvest had higher HIB antibody concentrations during the first 2 years post-transplant, the differences reaching significance at 3 months (P = 0.0001) and following the 24-month dose (P = 0.048) . Tetanus toxoid antibody concentrations were also significantly higher at 3 months (P = 0.001) and 6 months (P = 0.032) in patients immunized before BM harvest . There were no differences in pneumococcal antibody concentrations between the two groups . Immunization of patients before bone marrow harvest resulted in higher anti-HIB antibody concentrations following ABMT and may be an effective strategy to prevent infectious complications. Int J Pediatr Otorhinolaryngol, 1996 Jun, 36(1), 1 - 12 Effect of Haemophilus influenzae type b conjugate vaccine in combination with peroral immunization with Escherichia coli on experimental otitis media; Melhus A et al.; The protective ability of a conjugated Haemophilus influenzae type b vaccine, ACT-HIB, used singly or in combination with orally administered Escherichia coli, was investigated in a rat model for acute otitis media . The humoral response to ACT-HIB was also analyzed . The study demonstrated that ACT-HIB vaccination resulted in a prompt antibody response, and that ACT-HIB was efficient in preventing middle ear infections caused by Haemophilus influenzae type b . The efficiency increased if the vaccine was combined with Escherichia coli . The results suggest that Escherichia coli could possibly be useful in the future as a vaccine vehicle, and since Haemophilus influenzae acute mastoiditis seems to be almost exclusively due to serotype b, the incidence of this infection may be reduced with the conjugated Haemophilus influenzae type b vaccines. Ann Trop Paediatr, 1996 Jun, 16(2), 169 - 72 Chloramphenicol-resistant bacterial meningitis in Malawi; Graham SM et al.; Three recent cases of chloramphenicol-resistant pyogenic meningitis are reported from Malawi . The implications of the emergence of antibiotic-resistant strains of Streptococcus pneumoniae and Haemophilus influenzae type b causing childhood meningitis are discussed. Ann Trop Paediatr, 1996 Jun, 16(2), 103 - 11 Haemophilus influenzae type b disease in the western region of The Gambia: background surveillance for a vaccine efficacy trial; Adegbola RA et al.; In preparation for a field trial of an Haemophilus influenzae type b conjugate vaccine in the Western Region of The Gambia, a 3-year prospective study was undertaken to determine the incidence of Hib disease and the vaccination status of affected children . One hundred and eighty-two children with invasive Hib disease were found; 141 (77%) had meningitis, 31 (17%) pneumonia and 10 (6%) other forms of invasive disease . The estimated annual incidence rates for all invasive Hib diseases were 274 and 73 per 100,000 in children aged < 1 and < 5 years, respectively . For meningitis, the rate was 222 per 100,000 per year in children aged < 1 year . Children with meningitis were significantly younger than those with pneumonia (median age 7 months, interquartile range {IQR} 5-9, vs 12 months, IQR 6-15 (P = 0.002)) and younger than those with other forms of Hib disease . Of 142 children for whom vaccination status was known, 18 had received no DPT, 36 had received one, 40 had received two and 48 had received three doses . This study confirmed the high incidence of systemic Hib disease among Gambian children and the need to vaccinate at an early age . It provided the background epidemiological data required for the successful planning of an Hib vaccine trial which is now in progress. Clin Infect Dis, 1996 Jun, 22(6), 1069 - 76 Invasive disease due to Haemophilus influenzae serotype f: clinical and epidemiologic characteristics in the H . influe