Haemophilus

Commun Dis Intell, 1997 Feb 20, 21(4), 41 - 6
Epidemiology and prevention of pneumococcal disease; McIntyre P; There are comparatively little data on the incidence and morbidity from pneumococcal disease in Australia and elsewhere . Available data suggest that the overall incidence of invasive pneumococcal disease in Australia is comparable with similar populations . Very high rates are reported in Central Australian Aborigines, similar to invasive Haemophilus influenzae type b (Hib) disease . Disease incidence is probably greatly underestimated by case ascertainment from sterile site isolates alone . New diagnostic methods, such as serology to detect components of the pneumococcal cell wall, promise to significantly enhance detection of pneumococci as a cause of pneumonia, especially in childhood, but are epidemiologic rather than clinical tools . Resistance to penicillin and other antibiotics is an increasing problem worldwide, promoted by excessive antibiotic use, especially in children . This has focused attention on vaccine prevention . Fortunately, antibiotic-resistant pneumococci appear to belong to a limited range of serotypes, those commonly colonising children, in all areas so far studied . If conjugate pneumococcal vaccines prove to eradicate carriage, in a similar fashion to conjugate Hib vaccines, vaccination may be the major weapon against the spread of antibiotic-resistant pneumococcal infection . Conjugate pneumococcal vaccines are now in large scale efficacy trials, with outcomes of bacteraemia (California) and otitis media (Finland) . Results of these trials are eagerly awaited.

FEMS Microbiol Lett, 1997 Feb 15, 147(2), 233 - 8
Characterization of a novel transporter family that includes multiple Escherichia coli gluconate transporters and their homologues; Peekhaus N et al.; The nucleotide sequences of seven Escherichia coli genes that encode members of the gluconate permease (GntP) family have recently become available . These genes include gntP, gntU, gntW, ORf449, dsdX, and ORFo454 . The deduced amino acid sequences of all seven E . coli genes are homologous to the gntP gene products from Bacillus subtilis and B . licheniformis as well as two additional gene products from Haemophilus influenzae . These 11 proteins are not demonstrably homologous to members of the major facilitator superfamily or other recognized permease families . Four of the E . coli gluconate transporter genes have been cloned and shown to encode gluconate transporters with apparent affinities ranging from 6 to 212 microM . These studies serve to characterize a novel family of bacterial permeases.

MMWR Morb Mortal Wkly Rep, 1997 Feb 7, 46(5), 107 - 9
FDA approval for infants of a Haemophilus influenzae type b conjugate and hepatitis B (recombinant) combined vaccine.
{Acute pericarditis in childhood . The 9-year experience of a tertiary referral center}
Farinha NJ, Bartolo A, Trindade L, Vaz T, Monterroso J, Areias JC, Santos NT.

Servico de Cardiologia Pediatrica, Departamento de Pediatria, Hospital S . Joao, PortoSixteen children, aged 7 months to 12 years, with acute pericarditis, admitted between 1985 and 1993 to a tertiary referral centre were analyzed retrospectively for their presentation, etiology, work-up, management and prognosis . It was found that most of the presenting signs were not specific and were often related to associated diseases such as respiratory tract infections . In 50% of the cases a cause was not found, the others had viral infections (12.5%), tuberculosis (12.5%), Haemophilus influenzae infection (6.25%), Toxocara canis infection (6.25%) and collagen diseases (12.5%) . In eight cases non-steroidal anti-inflammatory drugs associated with steroids were given, 7 patients received non-steroidal anti-inflammatory drugs and 1 steroids . The mean follow-up time was 3 years (1 to 5) . Six patients had one or more relapses . Five of the 6 patients with relapses were in the group which received steroids . The two patients with tuberculosis underwent pericardiectomy . One child died due to complex heart disease and the remaining 15 were cured . It was concluded that in pericarditis an extensive work-up may not reveal the major etiologies and that long term prognosis is good.

Acta Med Port, 1997 Feb-Mar, 10(2-3), 129 - 38
{Anti-hepatitis B and anti-Haemophilus influenzae type b vaccines}; Dias PG; The epidemiologic aspects of hepatitis B are analysed . The author proposes the vaccination of all children, risk groups and teenagers . Also the high incidence of Haemophilus influenzae infections in Portugal justifies the use of an anti-Hib vaccine administered to all children under 5 years, preferentially in the first year of life.

Pneumologie, 1997 Feb, 51(2), 99 - 103
{Differential cytology of bronchoalveolar lavage fluid in healthy children in comparison with children colonized by lung-specific pathogens}; Ahrens P et al.; BAL is increasingly used in the assessment of pulmonary diseases, however, the role in children remains to be defined . Therefore bronchoalveolar lavage (BAL) was performed on 21 healthy children in order to enable quantification and characterization of the alveolar cell populations . Reference values for cellular constituents of BAL fluid using rigid bronchoscopes are lacking-even though in children still 90% of bronchoscopies are done by rigid instruments and BAL is performed using a catheter leading into the lower lobe . The children, 12 months to 14 years of age, were lavaged adjusted to body weight with three 1 ml/kg aliquots of normal saline . Recovery was 31 +/- 12% . Each aliquot was analysed separately . The number of total cell count per milliliter was 29.8 x 10(4) (range 0.6-161 x 10(4) . Percentages of different cell types of the third simple were (mean): macrophages 88.5, lymphocytes 6.4, neutrophils 4.8, eosinophils 0.1, basophils 0% . Significantly more neutrophils and less lymphocytes were found in the first as compared to the second and third sample . No age dependency was observed for either cell type . The comparison to cellularity of BAL-fluid with proven evidence of Adenovirus, Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae type B showed a marked influence of viral and bacterial germs . The study provides the first reference data on BAL differential cytology in healthy children using rigid bronchoscopy.

Oral Microbiol Immunol, 1997 Feb, 12(1), 27 - 32
Myeloperoxidase isoform activities released by human neutrophils in response to dental and periodontal bacteria; Miyasaki KT et al.; Myeloperoxidase is an enzyme released by polymorphonuclear leukocytes which has been used, experimentally, as an indicator of periodontal disease activity when measured in gingival crevicular fluid . There are three myeloperoxidase isoforms: MPO I, MPO II and MPO III . We examined the activities of myeloperoxidase isoforms released by neutrophils in response to serum-opsonized Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, Eikenella corrodens, Capnocytophaga sputigena and Streptococcus sanguis . Isoform activities were determined using intermediate-pressure liquid chromatography and microenzyme assay . A . actinomycetemcomitans stimulated higher levels of myeloperoxidase release than any other oral bacteria unless pre-opsonized with serum (or protein-A-purified immunoglobulin) from an individual with localized juvenile periodontitis . Most oral bacteria stimulated the release of all myeloperoxidase isoforms with a profile enriched in MPO I and diminished in MPO III . Exceptionally, serum-opsonized A . actinomycetemcomitans stimulated myeloperoxidase isoform release in proportion to the neutrophil granule constituency with or without localized juvenile periodontitis serum pre-opsonization . Because myeloperoxidase isoform profiles reflect how neutrophils were stimulated, isoform analysis may refine future diagnostic tests based upon myeloperoxidase.

Vaccine, 1997 Feb, 15(3), 325 - 8
Factors associated with superior antibody responses to a single dose of Haemophilus influenzae type b-tetanus toxoid conjugate vaccine administered to Chilean infants at 2 months of age; Levine OS et al.; The anticapsular antibody response of Chilean infants to a single dose of Haemophilus influenzae type b capsular polysaccharide-tetanus toxoid conjugate, vaccine is substantially higher than that observed among infants of similar age from the USA . Comparison of selected demographic and environmental factors indicates that low maternal education and a greater number of persons in the home are significantly associated with the superior responder phenotype . High anticapsular antibody responses were associated with high antibody responses to tetanus toxoid, the carrier protein in this conjugate, but not to diphtheria toxoid . These data suggest that environmental factors may enhance the magnitude of the primary antibody response to PRP-T vaccine.

Vaccine, 1997 Feb, 15(3), 295 - 300
Safety and immunogenicity of an acellular pertussis diphtheria tetanus vaccine given as a single injection with Haemophilus influenzae b conjugate vaccine; Halperin SA et al.; To determine if an acellular pertussis-diphtheria-tetanus vaccine could be combined with a Haemophilus influenzae b conjugate vaccine as a single injection, we randomized 468 children between 17 and 21 months of age previously immunized with three doses of each vaccine to receive a five-component acellular pertussis vaccine combined with diphtheria and tetanus toxoids, and Haemophilus influenzae b-tetanus toxoid conjugate vaccine either as separate injections on separate days, separate injections on the same day, or as a single combined injection . Rates of adverse reactions were similar amongst the groups except for increased injection site tenderness (29.2% vs 17%, RR 1.76, 95% CI 1.09-2.85) and fussiness (36.4% vs 23.3%, RR 1.59, 95% CI 1.05-2.41) in the group given the combined injection rather than separate injections on separate days . Antibody levels against the capsular polysaccharide of H . influenzae b after the single combined injection (47.1 micrograms ml-1) were lower than after separate injections on the same day (66.0 micrograms ml-1; P < 0.05) but higher than when the injections were administered on separate days (28.4 micrograms ml-1; P < 0.001) . We conclude that the five-component acellular pertussis vaccine is safe and immunogenic when combined with diphtheria and tetanus toxoids and H . influenzae b-tetanus toxoid conjugate vaccine in children receiving the fourth dose of the immunization series.

Acta Paediatr Jpn, 1997 Feb, 39(1), 44 - 7
Rapid, sensitive and specific diagnosis of Bordetella pertussis using the polymerase chain reaction; Aoyama T et al.; Use of a repetitive DNA sequence of Bordetella pertussis allowed successful detection of the organism by the polymerase chain reaction (PCR) . The method was highly sensitive, being able to detect B . pertussis in specimens containing only a few cells . It was also highly specific, with no amplification of specimens containing other organisms, for example Haemophilus influenzae or Neisseria, being observed . A diagnosis could be made within 1 day . The PCR assay was also evaluated in clinical specimens . Among 47 nasopharyngeal specimens obtained from 24 patients with laboratory-confirmed pertussis, 27 were positive by PCR and 19 by culture . In particular, all three bronchial aspirates from one patient with pertussis were positive by PCR, but only one showed positive on culture . Eleven specimens from parapertussis patients and 65 specimens from patients without pertussis tested negative . It was concluded that this newly developed PCR method for the diagnosis of pertussis was more rapid and sensitive than the usual culture method . Polymerase chain reaction could have a major impact on the treatment and control of this infection and would be a useful tool for studying the pathogenesis of B . pertussis infection.

Sex Transm Dis, 1997 Feb, 24(2), 64 - 8
Iron and virulence of Haemophilus ducreyi in a primate model; Sturm AW; BACKGROUND AND OBJECTIVES: The pathogenesis of chancroid is currently unclear . There are discrepancies between the virulence of Haemophilus ducreyi in the human and animal experiments . Iron availability influences the virulence of many pathogens . The objective of this study was to investigate the role of iron in virulence of H . ducreyi . STUDY DESIGN: Intradermal inoculation of the primate Macaca mulatta was studied as an animal model for pathogenesis of chancroid . This model then was used to study the influence of iron availability on virulence of H . ducreyi . Eleven strains of H . ducreyi with proven avirulence in the rabbit intradermal test and one virulent control strain were used . Two of the strains had been shown to cause ulcers in a human inoculation experiment . Strains were suspended in fluid with different iron compounds before intradermal injection . Other monkeys were treated with injectable iron before inoculation with bacteria suspended . RESULTS: All but one of the test strains produced a chancroid-like ulcer from which H . ducreyi could be isolated . The minimal ulcerative dose was 10(5) cfu . Iron in the injection fluid decreased this dose 10-fold . In animals pretreated with intramuscular iron, the minimal ulcerative dose decreased to 10(4) cfu and all eleven strains became ulcerative . CONCLUSIONS: The availability of iron increases virulence of H . ducreyi . The iron pretreated primate model is useful for the study of virulence factors of H . ducreyi because of its ability to produce lesions with strains that avirulent in other animal models but pathogenic to humans.

J Chemother, 1997 Feb, 9(1), 44 - 50
Comparative trial of 3 days of azithromycin versus 10 days of clarithromycin in the treatment of children with acute otitis media with effusion; Arguedas A et al.; The authors compared the efficacy, safety and tolerance of azithromycin and clarithromycin in pediatric patients with acute otitis media . A randomized, open clinical trial was performed comparing azithromycin and clarithromycin in children aged 6 months to 12 years of age with acute otitis media with effusion . Patients were allocated to azithromycin at 10 mg/kg once daily for 3 days or to clarithromycin at 15 mg/kg day divided into two equal doses for 10 days . Clinical examinations and tympanometric evaluations were performed at baseline, day 3-5, day 10-14, day 28-30 and day 50-60 . Tympanocentesis fluid cultures were collected at enrollment and urine and blood samples were obtained at baseline and day 10-14 . Of 100 patients enrolled, 97 were considered evaluable . The most common middle ear pathogens were Streptococcus pneumoniae (60%), Haemophilus influenzae (15%) and Staphylococcus aureus (13%) . Fifty patients (100%) treated with azithromycin and 45 (95.7%) patients treated with clarithromycin had a satisfactory clinical response . Rates of persistence of middle ear effusion and possible drug related side effects were comparable . Based on the efficacy and safety results, azithromycin for 3 days and clarithromycin for 10 days are considered to represent an attractive alternative for the treatment of children with acute otitis media.

J Antibiot (Tokyo), 1997 Feb, 50(2), 162 - 8
Structure-activity relationships of carbapenem compounds to anti-Haemophilus influenzae activity and affinity for penicillin-binding proteins . Effect of 1 beta-methyl group and C-2 side chain; Kanazawa K et al.; The anti-H . influenzae activity of meropenem (1a) was much higher than those of imipenem (4) . panipenem (2b) and biapenem (7) . To clarify the major structural features responsible for the anti-H . influenzae activity of carbapenem compounds, the structure-activity relationship to the anti-H . influenzae activity was investigated . The anti-H . influenzae activities of meropenem (1a) and 1 beta-methyl-panipenem (2a) were much higher than those of desmethyl-meropenem (1b) and panipenem (2b) . respectively . Two carbapenems (5, 6) and imipenem (4), that have a strong basic C-2 side chain, showed lower anti-H . influenzae activity than meropenem (1a) having a weakly basic C-2 side chain and N-acetyl thienamycin (3) having a neutral C-2 side chain, respectively . As a result, we found that the introduction of the 1 beta-methyl group or the reduction of the basicity (cationic character) of the C-2 side chain increased the antimicrobial activity and bactericidal activity of carbapenems against H . influenzae due to their increased affinity for PBP-4 and PBP-5.

Pathology, 1997 Feb, 29(1), 72 - 5
In vitro susceptibility patterns of nonserotypable Haemophilus influenzae from patients with chronic bronchitis; Butt HL et al.; The antimicrobial susceptibility patterns of 76 nonserotypable Haemophilus influenzae (biotypes I-IV) from patients with chronic bronchitis were compared against ten orally administered antimicrobial agents . In addition the sputum ampicillin concentrations one hour after standard therapy were determined in five patients with chronic bronchitis . Ampicillin resistance was demonstrated in one strain (biotype IV) which produced beta-lactamase and two strains (biotype II) with innate resistance (MIC = 4 mg/l) . Resistance to trimethoprim, chloramphenicol, ciprofloxacin and cefaclor was not detected . The incidence of resistance to tetracycline was 0.5% and cephalexin 13.2% . A high incidence of resistance to erythromycin (95%) was noted . There was no association between resistance and biotype of nonserotypable H . influenzae . The sputum ampicillin concentrations from four out of five patients given standard antibiotic doses were shown to be sufficient to inhibit the growth of the majority of nonserotypable H . influenzae strains one hour after treatment . This study shows that the incidence of nonserotypable H . influenzae resistant to ampicillin is low in this community but that resistance levels to erythromycin, commonly prescribed for the management of acute bronchitis, are high . Regular sensitivity screens are important in monitoring the value of various antibiotic regimens in the management of acute bronchitis.

Aust N Z J Med, 1997 Feb, 27(1), 59 - 61
Community acquired pneumonia in north eastern Australia--a hospital based study of aboriginal and non-aboriginal patients; Thompson JE; BACKGROUND: Although the most important organism causing community acquired pneumonia continues to be Streptococcus pneumoniae, the frequency of other pathogens varies considerably across the world . AIMS: To look for the causes of community acquired pneumonia (CAP) in north east Australia . To examine the differences between Aboriginal and non-Aboriginal subjects with emphasis on prevalence, risk factors, causative organisms and prognosis . METHODS: The records of 200 consecutive patients admitted to Cairns Hospital during 1992 with a diagnosis of pneumonia were studied . RESULTS: Aboriginal patients made up half the numbers admitted, but only 13% of the population . More Aboriginal adults died than non-Aboriginal adults, but no child died . Heavy alcohol use, chronic lung disease and diabetes mellitus were the commonest risk factors for Aborigines (alcohol, chronic lung disease and cancer for non-Aboriginal subjects) . S . pneumoniae, followed by Haemophilus influenzae were the most common pathogens found . Both were sensitive to amoxycillin . The yield of definite pathogens was low . Staphylococcal and melioidosis pneumonia were confined to Aboriginal patients . Chlamydia, legionella and Mycoplasma pneumoniae were not identified.

J Trop Pediatr, 1997 Feb, 43(1), 10 - 2
Spectrum of Haemophilus influenzae type b disease in children at a university hospital in Riyadh, Saudi Arabia; Abdullah AM et al.; During a period of 5 years, 42 cases of invasive Haemophilus influenzae type b (Hib) disease were studied . An outstanding finding in our patients was occult bacteraemia, which was detected in 13 (31 per cent) patients . Other clinical conditions encountered were nine (21 per cent) patients with pneumonia, nine (21 per cent) with meningitis, six (14 per cent) with cellulitis, three (7 per cent) with arthritis, one (2 per cent) with epiglottitis, and one (2 per cent) with urinary tract infection . The mean age of children was 21 months (range 1-156 months); the majority (62 per cent) belonged to the age group 7-18 months . There were 24 females and 18 males . Eighty-one per cent of these patients were Saudi nationals . Five isolates (12 per cent) of Hib were resistant to ampicillin and similar numbers were resistant to chlorampenicol . Twenty-five children (60 per cent) were treated with ampicillin, nine (21 per cent) with chlorampenicol and eight (19 per cent) with ceftriaxone . All patients made complete recovery.

J Formos Med Assoc, 1997 Feb, 96(2), 110 - 5
Safety and immunogenicity of a conjugated Haemophilus influenzae type B polysaccharide-Neisseria meningitidis outer membrane protein vaccine; Yeh SJ et al.; This study was performed to determine the immunogenicity and safety of a conjugate Haemophilus influenzae type b (Hib) vaccine (PedvaxHIB:Merck Sharp & Dohme Inc, West Point, PA, USA) in Taiwanese children . There were two groups in this study . Group A included 56 infants at 2 months of age, who received an initial dose of Hib conjugate vaccine at age 2 months and 4 months . Another booster dose was given between 12 and 15 months of age . Group B included 58 children, aged 11 to 14 months, who were vaccinated at the time of enrollment and 2 months after the first vaccination . The results showed that this Hib conjugate vaccine was highly immunogenic with 100% and 86% of vaccinees in group A developing anti-Hib polyribosylribitol phosphate (PRP) antibody titers above 0.15 microgram/mL and 1 microgram/mL, respectively, after two vaccinations . A slightly better response rate was observed in group B, with 100% and 97% of vaccinees developing antibody titers above 0.15 microgram/mL and 1 microgram/ml, respectively, after two vaccinations . A significant booster effect was seen in group A; the proportion of subjects with antibody titers above 1 microgram/mL increased from 38% to 95%, and the geometric mean titer increased from 0.68 microgram/mL to 11.92 micrograms/mL . Adverse reactions were uncommon . Mild reactions consisted of low-grade fever and local discomfort . We conclude that the Hib conjugate vaccine is highly immunogenic and safe when given to Taiwanese children as young as 2 months of age.

Enferm Infecc Microbiol Clin, 1997 Feb, 15(2), 65 - 9
{Maternal-fetal vertical transmission of Haemophilus influenzae and Haemophilus parainfluenzae}; Garcia E et al.; BACKGROUND: Maternal-fetal transmission of Haemophilus influenzae (HI) and Haemophilus parainfluenzae (HPI) as well as characteristics of the infection in the neonate have been poorly described . METHODS AND RESULTS: From 1982 to 1994 18 HI and 12 HPI from vaginal secretion of 30 women obtained intrapartum or immediately post partum were isolated . The following was observed in the 18 patients in whom HI was isolated: 1 abortion, 2 antepartum death, 1 post partum death, and 14 live neonates from whom 10 were suspect a infection with 5 cases being confirmed with isolation of HI in blood culture . Mortality was 40% . The rate of vertical transmission of HI was 80% (8/10) and neonatal sepsis 50% (5/10) (71.4% of the infants whose mothers did not receive intrapartum antibiotics and 0% in those who did) . Twenty-seven point seven percent of the strains of HI were beta-lactamase producers with II and III being the most frequent biotypes observed . Of the 12 patients from whom HPI was isolated the following was reported: 1 antepartum death, and 12 live neonates of whom 10 were suspect a infection, but no case was confirmed because HPI was not isolated from blood culture . The vertical transmission of HPI was 20% (2/10) . CONCLUSIONS: The isolation of HI in the vagina of pregnant women should be considered as being potentially very virulent since despite a low prevalence, the rate of vertical transmission and, overall, neonatal sepsis, is high.

J Antimicrob Chemother, 1997 Feb, 39(2), 209 - 16
Anti-CD11b monoclonal antibody in an infant rat model of Haemophilus influenzae type b sepsis and meningitis; Tan TQ et al.; Five-day-old infant rats were injected intraperitoneally (i.p.) with anti-CD11b monoclonal antibody (1 B6) at a dose of 2 mg/kg or phosphate-buffered saline (PBS) either 1 h before or 3 or 24 h after inoculation with 10(5) cfu Haemophilus influenzae type b (Hib) . When administered 1 h before infection, 23% of the 1B6- versus 17% of the PBS-treated rats and 87% of the 1B6- versus 83% of the PBS-treated animals died at 24 and 48 h, respectively . There was a similar mortality for 1B6 or PBS treatment at 3 h after infection . Thirteen of 15 (87%) 1B6 animals versus 16/17 (94%) PBS animals had positive CSF cultures at 48 h . No differences in mortality were observed in separate experiments where animals received 1B6 or PBS 3 or 24 h after infection with Hib and were treated with a single ampicillin dose (100 mg/kg) 24 h after infection . The median CSF white blood cell count/mm3 was 5627 and 4860 for the animals with meningitis receiving 1B6 and PBS, respectively, although the 1B6-treated animals had a lower percentage of polymorphonuclear cells in the CSF (P = 0.05) . Histologic examination of the meninges, choroid plexus and cochlea showed a slight decrease in the numbers of inflammatory cells in animals treated with 1B6 . 1B6 did not change the incidence of meningitis and only slightly decreased the degree of inflammation within the central nervous system, although animals treated with 1B6 have an altered CSF leucocyte response with the presence of more mononuclear cells as opposed to polymorphonuclear cells in their CSF . 1B6 may play a role in inhibiting neutrophil emigration to sites of inflammation within the central nervous system but is not beneficial in decreasing mortality in an infant rat model of H . influenzae type b sepsis and meningitis.

Arch Dis Child, 1997 Feb, 76(2), 129 - 33
Evaluation of a protocol for selective empiric treatment of fever without localising signs; Browne GJ et al.; A protocol for management of young febrile children at risk for bacteraemia has been used at Westmead Hospital, a university based hospital in the western Sydney region, since early 1994 . Implementation of the protocol was retrospectively evaluated for the 12 month period 1 June 1994 to 31 May 1995, using the emergency department log book as the primary data source . Altogether 498 children, aged from 3 months to 3 years, with a fever > or = 39.5 degrees C were identified over this period, of whom 291 were admitted to hospital because of evidence of sepsis or identified focal infection and 207 children without focal infection were observed in the short stay annexe of the emergency department . Fifty children, considered at high risk of bacteraemia because of a total white cell count > or = 20 x 10(9)/1 received empiric antibiotic treatment with ceftriaxone, of whom 19 subsequently had proved bacteraemia and another 10 had focal infection identified during observation in the short stay annexe . Bacteraemia was due to Streptococcus pneumoniae in 16 cases and Haemophilus influenzae type b in three . No adverse events occurred at follow up . Use of a management protocol and selection on higher white cell count criterion than previously recommended by US centres resulted in restriction of empiric antibiotic treatment to a small proportion of young febrile children presenting to a busy emergency department of whom 38% were bacteraemic.

Vaccine, 1997 Feb, 15(2), 209 - 19
Persistence of antibodies to meningococcal IgA1 protease versus decay of antibodies to group A polysaccharide and Opc protein; Thiesen B et al.; Sera were taken over a 5 year period from Gambian children vaccinated in 1983, when aged 1-4 years, with A + C meningococcal capsular polysaccharide, ELISA tests were devised to determine the concentrations of immunoglobulin A, G and M reacting with A polysaccharide and of IgG reacting with Opc protein, IgA1 protease and an internal 104 mer peptide derived from IgA1 protease . Vaccination resulted in a brief rise of antibodies to A polysaccharide followed by decline to pre-immunization levels . IgM levels were very high even before vaccination . Antibodies to Opc protein stimulated by natural exposure also declined over the 5 year period . In contrast, antibodies stimulated by natural exposure to IgA1 protease or to the internal peptide remained constant or increased (final geometric mean level of 47 micrograms IgG ml-1) . We speculate that healthy carriage of Neisseria meningitidis or Haemophilus influenzae is responsible for this increase in IgG concentration.

Vaccine, 1997 Feb, 15(2), 149 - 54
Immunogenicity and safety of a liquid combination of DTP-PRP-T {corrected} vs lyophilized PRP-T reconstituted with DTP; Amir J et al.; The immunogenicity and safety of a combined diphtheria, tetanus, pertussis and Haemophilus influenzae type b-tetanus conjugate vaccine (DTP-PRP-T) was compared to the same combination obtained by the reconstitution of H . influenzae type b-tetanus conjugate vaccine lyophilized (PRP-T) with liquid diphtheria-tetanus-pertussis vaccine (DTP) . Two hundred and sixty-two healthy infants were randomized to receive a intramuscular injection of 0.5 ml of one of the above combination vaccines at 2, 4 and 6 months of age, and a subgroup of 134 infants received a booster dose at 12 months . Serum antibody levels to each vaccine component were measured at ages 2, 6, 7, 12 and 13 months . Systemic and local reactions were assessed during the first 3 days after each injection by diary cards distributed to the parents . After the third dose and booster administered at 12 months of age, significant equivalence between the groups was observed, and the geometric mean titer of anti H . influenzae type b capsular polysaccharide (Hib-CP) antibodies were 5.9 and 32.6 micrograms ml-1 for the liquid combination group and 5.8 and 19.4 for the lyophilized group, respectively . After the third dose, anti-Hib-PC antibody levels of > or = 1.0 microgram ml-1 and 0.15 microgram ml-1 were seen in 94% and 100%, respectively, of the liquid combination group and 90 and 99%, respectively of the lyophilized group . After the booster dose, levels of > or = 1.0 microgram ml-1 were observed in 100% and 93.5% of the liquid combination group and the lyophilized combination group, respectively . Systemic and local reactions to the vaccination were generally mild and did not differ significantly between the groups . We conclude that the liquid combination of DTP-PRP-T is safe and at least as immunogenic as the lyophilized preparation . This liquid preparation, like other combined vaccines may be helpful for planning vaccination programs with a reduced number of injections.

Pediatr Emerg Care, 1997 Feb, 13(1), 16 - 8
Aerobic and anaerobic microbiology of bacterial tracheitis in children; Brook I; STUDY OBJECTIVES: Establish the aerobic and anaerobic microbiology of bacterial tracheitis in children . METHOD: Retrospective review of specimens obtained from 14 children with bacterial tracheitis that were cultured for aerobic and anaerobic bacteria . RESULTS: A total 30 bacterial isolates were recovered, 17 aerobic and facultative anaerobic and 13 anaerobic . Aerobic bacteria only were present in six (43%) specimens, anaerobes only in three (21%), and mixed aerobic and anaerobic flora in five (36%) . Polymicrobial flora was recovered in 10 of the 14 specimens . The predominant organisms were Staphylococcus aureus (5 isolates), Haemophilus influenzae type b (4), Peptostreptococcus sp . (4), pigmented Prevotella and Porphyromonas (4), Fusobacterium sp . (2), and Moraxella catarrhalis (2) . Two organisms that were also isolated from the tracheal aspirates were recovered from the blood of two patients (one each of H . influenzae and Prevotella intermedia) . Eleven beta-lactamase-producing organisms were isolated from nine patients . These included all isolates of S . aureus and M . catarrhalis, and two each of H . influenzae and Prevotella sp . CONCLUSIONS: These data confirm the predominance of S . aureus and H . influenzae in causing bacterial tracheitis in children and suggest a potential role for anaerobic bacteria.

Acta Med Okayama, 1997 Feb, 51(1), 33 - 7
Clinical effect of low-dose, long-term roxithromycin chemotherapy in patients with chronic sinusitis; Kimura N et al.; We studied the clinical efficacy of roxithromycin (RXM) administered at the daily dosage of one tablet (150 mg) for 3 months in 30 patients with chronic sinusitis . The effectiveness of this drug was evaluated on a four-point scale . Subjective and objective symptoms disappeared or decreased markedly, especially postnasal drip and nature of discharge in 80 percent or more of the patients . All symptoms significantly decreased (P < 0.001; headache P < 0.05), except for the sensation of foul odor . Symptoms improved even in those cases in which Haemophilus influenzae was detected . It is suggested that RXM produce some clinically beneficial effect through an immunological and or anti-inflammatory mechanisms in addition to its antibiotic effect.

Eur J Biochem, 1997 Feb 1, 243(3), 701 - 7
Structural studies of the cell-envelope oligosaccharide from the lipopolysaccharide of Haemophilus influenzae strain RM.118-28; Risberg A et al.; The structure of the oligosaccharide part of the Haemophilus influenzae RM.118-28 lipopolysaccharide (LPS) has been investigated . The oligosaccharide was obtained from the LPS by mild acid hydrolysis followed by gel-permeation chromatography, and was studied by methylation analysis, NMR spectroscopy and mass spectrometry . The structure of the major compound, which is a hexasaccharide, is proposed as follows . {formula: see text} In the structure, Kdo is 3-deoxy-D-manno-octulosonic acid, PEtn is phosphoethanolamine, PCho is phosphocholine and L,D-Hep is L-glycero-D-manno-heptose . Electrospray-ionization mass spectrometry on O-deacylated LPS obtained after treatment with anhydrous hydrazine gave evidence for the presence of two minor compounds, which show additional substitution of the main structure with phosphate and PEtn, respectively . These substitutions have not been localized.

Otolaryngol Head Neck Surg, 1997 Feb, 116(2), 175 - 80
Middle ear mucin glycoprotein: purification and interaction with nontypable Haemophilus influenzae and Moraxella catarrhalis; Reddy MS et al.; Nontypable Haemophilus influenzae and Moraxella catarrhalis are important pathogens in children and adults . The mechanisms of their adherence to the epithelial cell surface and colonization are not clear . For the pathogen to adhere to the epithelial cell, it must first attach to and penetrate the mucus barrier . Mucin glycoproteins of the mucus layer generally are thought to be involved in bacterial attachment . To understand the precise mechanisms of middle ear mucin-bacterial interactions, we used an overlay binding assay with a highly purified middle ear mucin and outer membrane proteins of both nontypable H . influenzae and M . catarrhalis . Outer membrane proteins P2 and P5 were identified as the major components that medicate the binding between nontypable H . influenzae and human middle ear mucin . Moreover, the 57 kDa protein, CD, of the outer membrane protein of M . catarrhalis was found to be the only protein binding human middle ear mucin . Finally, it appears that a protein-oligosaccharide interaction is responsible for binding because asialo-mucin does not bind to either of the bacteria . Knowledge of the specific bacterial-mucin interaction may provide an understanding of the bacterial-epithelial cell colonization . Conversely, comprehension of this interaction between bacteria and purified mucin may be a strategy to prevent colonization of potential pathogens that cause otitis media and sinusitis in children.

Postgrad Med, 1997 Feb, 101(2), 191 - 2, 195-6, 199-200
Haemophilus influenzae infections in adults: a pathogen in search of respect; Strausbaugh LJ; Despite the success of Haemophilus influenzae type b vaccines in preventing bacterial disease in children, H influenzae remains a common pathogen in adult patients in the United States and Europe . At least half of invasive H influenzae infections are caused by nontypable strains . The spectrum of diseases includes sinusitis, pneumonia, otitis media, epiglotitis, and meningitis . An etiologic diagnosis is most reliably established by positive cultures from a normally sterile site . Although resistance to ampicillin and amoxicillin has steadily increased in clinical H influenzae isolates during the past two decades, a variety of other antimicrobial agents are available for the treatment of infections caused by this bacterium.

Mol Microbiol, 1997 Feb, 23(3), 569 - 77
Characterization of monoclonal antibodies recognizing three distinct, phosphorylated carbohydrate epitopes in the lipopolysaccharide of the deep rough mutant I-69 Rd-/b+ of Haemophilus influenzae; Rozalski A et al.; Monoclonal antibodies against the lipopolysaccharide (LPS) of the deep rough mutant I-69 Rd-/b+ of Haemophilus influenzae were obtained after immunization of mice with sheep erythrocytes which had been coated with de-O-acylated LPS . Characterization of antibodies was performed by enzyme immuno assay (EIA) using LPS or neoglycoconjugates containing partial structures of LPS as solid-phase antigens and by haemagglutination with sheep erythrocytes coated with de-O-acylated LPS . Binding data were confirmed by EIA inhibition experiments using deacylated LPS or synthetic partial structures thereof . Three antibodies were specific for 3-deoxy-D-manno-octulopyranosonic acid- (Kdo) 5-phosphate, one for Kdo-4-phosphate, and one required, in addition to a Kdo-phosphate, parts of the phosphorylated glucosamine backbone of lipid A . All antibodies also bound in (i) Western blots to bacterial whole-cell lysates or isolated LPS separated by SDS-PAGE, (ii) bacterial colony blots, and (iii) immunofluorescence with live bacteria . The latter result indicated that Kdo-4- and Kdo-5-phosphate are synthesized by the bacteria and are not the result of phosphate migration.

Br J Oral Maxillofac Surg, 1997 Feb, 35(1), 43 - 5
Selective use of tracheostomy in surgery for head and neck cancer: an audit; Crosher R et al.; This retrospective study was designed to define the role of tracheostomy in the operative treatment of patients with cancers of the head and neck . The subjects were 51 patients who underwent neck dissection with resection of the tumour and repair during the period January 1992-December 1994, out of a total of 109 patients who were treated for cancers of the head and neck during that time . Three patients required tracheostomies, two of which were done preoperatively, and one immediately postoperatively for respiratory distress . There were no operative deaths . Morbidity included wound infection (n = 2), chest infection caused by Haemophilus influenzae (n = 1), transient fever associated with blood transfusion (n = 5), and transient fever of no obvious cause (n = 3) . Median hospital stay was 10 days (range 4-38) . Patients undergoing operations for cancers of the head and neck do not require routine tracheostomy . Further research on how to select patients who will need tracheostomy is necessary and is being done.

Pediatr Infect Dis J, 1997 Feb, 16(2 Suppl), S21 - 4
Comparison of the efficacy, safety and acceptability of cefixime and amoxicillin/clavulanate in acute otitis media; Gooch WM 3rd et al.; BACKGROUND: Use of a beta-lactamase stable antibiotic is called for in cases of acute otitis media (AOM) likely to be caused by beta-lactamase-producing Haemophilus influenzae or Moraxella catarrhalis . Two beta-lactamase-stable agents commonly used for empirical treatment of AOM are amoxicillin/clavulanate and cefixime . METHODS: A multicenter, randomized clinical trial compared cefixime (CFX; 8 mg/kg once daily) with amoxicillin/clavulanate (A/C; 40 mg/kg/day in three divided doses) for the treatment of children with AOM . Three hundred thirteen children were randomly assigned to a 10-day course of either CFX (n = 158) or A/C (n = 155) . Based on history, physical examinations and otoscopic and tympanometric assessments, clinical responses were evaluated as cure, improvement, failure, relapse or nonevaluable . Compliance and patient/parent acceptability were also analyzed . RESULTS: Overall favorable clinical responses (cure plus improvement) were comparable post-therapy for the two treatments (CFX = 76%; A/C = 77%) . Significant differences in response rates for both treatments were noted among different geographic regions, with the highest response rates observed in the Northeast and South . Acceptability of CFX was significantly better than that of A/C (P = 0.0001), and the adverse experience rate was lower (P = 0.001) . The most frequently reported adverse experiences were diarrhea (CFX 15.2%, A/C 29.7%) and vomiting (CFX 3.2%, A/C 10.32%) . Relapse rates were 26% for CFX and 29% for A/C . CONCLUSION: This study demonstrated that CFX has comparable clinical efficacy and a better adverse events profile than A/C when used to treat AOM of childhood.

Pediatr Infect Dis J, 1997 Feb, 16(2 Suppl), S17 - 20
Conjunctivitis in infants and children; Wald ER; BACKGROUND: Acute bacterial conjunctivitis, chiefly affecting young children, is generally caused by one of three common pathogens, Haemophilus influenzae, Streptococcus pneumoniae and adenovirus . H . influenzae is the most prevalent causative organism . Some patients presenting initially with conjunctivitis subsequently develop acute otitis media, with H . influenzae the most common etiologic agent of what has been termed the "conjunctivitis-otitis" syndrome . Optimal treatment of acute conjunctivitis would result in a clinical and bacteriologic cure of the conjunctivitis and the prevention of the development of otitis media . Application of topical antimicrobials at 4- to 6-h intervals a day for 1 week to 10 days results in the resolution of the conjunctivitis . However, topical treatment is difficult to administer to toddlers and does not effectively reduce the potential for the development of otitis media . Compared with topical and placebo treatment, oral antibiotics effective against H . influenzae have proved to be most effective in preventing the otitis media associated with conjunctivitis . CONCLUSION: Acute bacterial conjunctivitis is a common disorder in children < 6 years of age . H . influenzae is the most common etiology of the "conjunctivitis-otitis syndrome." The optimal treatment for this condition is still being studied.

Pediatr Infect Dis J, 1997 Feb, 16(2 Suppl), S12 - 6
Using antibiotic concentrations in middle ear fluid to predict potential clinical efficacy; Harrison CJ; BACKGROUND: The effectiveness of an antibiotic in eradicating an infection within an anatomic compartment is related to both its availability (penetration) and the susceptibility of the causative pathogen . Antimicrobial penetration can be assessed by measuring concentrations in serum or tissue . However, to assess antibiotic effectiveness in the treatment of otitis media, both antibiotic concentrations and pathogen profiles in middle ear fluid (MEF) should be considered . Tympanocentesis is required to access fluid in the middle ear compartment . This paper considers data on (MEF) antibiotic concentrations obtained with this procedure . METHODS: Data on MEF antibiotic concentrations were considered in relation to the antibiotic susceptibility profiles of the more common otitis media pathogens . RESULTS: Amoxicillin reaches MEF concentrations that exceed MICs for penicillin-susceptible and penicillin relatively resistant Streptococcus pneumoniae, but not for beta-lactamase-producing Haemophilus influenzae . Several beta-lactamase-stable drugs, e.g . cefixime and amoxicillin/clavulanate, however, achieve MEF concentrations that easily exceed their respective MIC90s for both beta-lactamase-producing and nonproducing H . influenzae but at ordinary doses may not exceed MIC90s for penicillin relatively resistant S . pneumoniae . CONCLUSION: Ongoing observation of organisms and their antibiotic resistance patterns in patients failing initial therapy of acute otitis media indicate that stability to beta-lactamase remains an essential quality for effective second line therapies . However, when possible tympanocentesis with culture is the ideal method of targeting specific therapy for patients failing multiple consecutive antibiotic regimens.

Pediatr Infect Dis J, 1997 Feb, 16(2 Suppl), S5 - 8
Role of nontypeable Haemophilus influenzae in pediatric respiratory tract infections; Klein JO; BACKGROUND: Nontypeable strains of Haemophilus influenzae may spread contiguously from the upper respiratory tract and cause sinusitis, otitis media and pneumonia . Unlike H . influenzae type b these strains rarely invade the bloodstream to cause widespread infections . These strains are primary pathogens of acute otitis media, sinusitis and the conjunctivitis-otitis syndrome . In developing countries these strains are also responsible for many cases of pediatric pneumonia . Currently approximately 30% of nontypeable H . influenzae strains are beta-lactamase-positive and can inactivate susceptible penicillins, including penicillins G and V, ampicillin and amoxicillin . Most second generation oral cephalosporins are active against beta-lactamase-producing H . influenzae . Some third generation oral cephalosporins, e.g . cefixime, however, have particularly good efficacy against H . influenzae . Sulfonamides and chloramphenicol are generally effective as well . CONCLUSION: Until a vaccine that provides adequate prophylaxis against nontypeable H . influenzae is developed, clinicians should be guided by knowledge of bacterial susceptibility and microbiologic efficacy in choosing appropriate agents for treating pediatric infections likely to involve this common pathogen.

FEMS Microbiol Lett, 1997 Feb 1, 147(1), 11 - 6
Cloning and characterization of the groE locus from Actinobacillus pleuropneumoniae; Vezina G et al.; A 4.4-kb DNA fragment was cloned from Actinobacillus pleuropneumoniae (strain 4074, serotype 1) by genetic complementation with Escherichia coli groES-groEL mutant strains . Sequence analysis of this fragment revealed a purine nucleoside phosphorylase (DeoD)-encoding gene homolog (deoD), heat-shock response-encoding genes for the small (groES) and large subunits (groEL) and a partial open reading frame encoding an alcohol dehydrogenase homolog (adhE) . The predicted amino-acid sequence of groES and groEL genes showed extensive sequence identity (80-95%) with other Pasteurellaceae . The gene organization surrounding the groE locus was different from that of Haemophilus infuenzae . When expressed in E . coli, groES-groEL genes were capable of complementing the growth of a lambda lytic phage, indicating a structural as well as functional conservation.

Am J Hematol, 1997 Feb, 54(2), 131 - 8
Infectious morbidity in long-term survivors of allogeneic marrow transplantation is associated with low CD4 T cell counts; Storek J et al.; Survivors of allogeneic marrow transplants are immunodeficient for at least 1 year after grafting . Multiple defects of immunity have been found; however, it is not known which defect primarily accounts for the high infectious morbidity of these patients . Twenty-nine allograft recipients who were in complete remission of the original disease were examined for the following parameters of immunity at 1 year after transplant: infection score (gauging the number and severity of infections within the 6 months prior to the annual exam), serum total IgM, IgG, and IgA, anti-Haemophilus influenzae IgG, anti-Streptococcus pneumoniae IgG, skin test reactivity, and the blood counts of B cells, CD4+ T cells, CD8+ T cells, and their subsets . THe only parameter inversely correlated with the infection score was CD4+ T cell count (P = 0.005 in univariable analysis, P = 0.06 in multivariable analysis) . We conclude that infectious morbidity of long-term transplant survivors is related to the reconstitution of CD4+ T cells.

Antimicrob Agents Chemother, 1997 Feb, 41(2), 454 - 9
In vitro evaluation of a novel ketolide antimicrobial agent, RU-64004; Jamjian C et al.; Ketolides, a novel macrolide subclass, possess a mode of action that is similar to that of structurally related macrolide-lincosamide-streptogramin (MLS) compounds . By using reference in vitro tests, the in vitro activity of RU-64004 was compared to those of six other MLS compounds against more than 800 clinical pathogens, including 356 gram-positive organisms . The spectrum of activity of the ketolide was most similar to that of clindamycin versus staphylococci and streptococci and superior to those of all macrolides tested against oxacillin-resistant staphylococci and vancomycin-resistant (vanA, vanB, and vanC) enterococcal isolates . The activity of the ketolide was greater than those of the macrolides, azalides, or clindamycin tested against vancomycin-susceptible enterococci (MICs at which 90% of isolates are inhibited {MIC90S}, 0.25 to 4 micrograms/ml), penicillin-resistant pneumococci (MIC90, 0.25 micrograms/ml), and most beta-hemolytic streptococci . All Streptococcus pneumoniae and beta-hemolytic streptococcus strain were inhibited by ketolide concentrations of < or = 0.25 micrograms/ml . Against 165 erythromycin-resistant strains, RU-64004 inhibited (MICs, < or = 0.5 micrograms/ml) approximately one-third of staphylococci, all streptococci, and slightly more than one-half of the enterococci . Quinupristin-dalfopristin (a streptogramin combination) was active against all tested isolates with the exception of non-Enterococcus faecium enterococci, against which the ketolide exhibited greater potency (MIC50S, 0.03 to 2 micrograms/ml) . The ketolide was also active against Haemophilus influenzae (MIC90, 2 micrograms/ml), Moraxella catarrhalis (MIC90, 0.12 micrograms/ml), pathogenic Neisseria spp . (MIC90, 0.5 micrograms/ml), and many gram-positive anaerobes (MIC90, 0.5 micrograms/ml) . RU-64004 may enhance the role of macrolide drugs in the treatment of some serious infections caused by MLS-resistant gram-positive organisms.

Antimicrob Agents Chemother, 1997 Feb, 41(2), 292 - 7
Antibiotic resistance among clinical isolates of Haemophilus influenzae in the United States in 1994 and 1995 and detection of beta-lactamase-positive strains resistant to amoxicillin-clavulanate: results of a national multicenter surveillance study; Doern GV et al.; A total of 1,537 clinical isolates of Haemophilus influenzae were recovered in 30 U.S . medical center laboratories between 1 November 1994 and 30 April 1995 and were characterized in a central laboratory with respect to serotype and beta-lactamase production and the in vitro activities of 15 oral antimicrobial agents . Overall, 36.4% of the isolates were found to produce beta-lactamase . The rank order of activity of six cephalosporins on the basis of MICs was cefixime > cefpodoxime > cefuroxime > loracarbef > or = cefaclor > cefprozil . On the basis of current National Committee for Clinical Laboratory Standards (NCCLS) breakpoints ages of isolates found to be resistant or intermediate to these agents were as follows: 0.1, 0.3, 6.4, 16.3, 18.3, and 29.8, respectively (National Committee for Clinical Laboratory Standards . Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically, 4th ed . M7-A4, 1995) . Azithromycin was, on a weight basis, the most potent of the macrolides tested in this study, followed by erythromycin and then clarithromycin . Azithromycin was typically fourfold more active than erythromycin, which was, in turn, slightly more active than clarithromycin . However, when compared on the basis of the frequency of resistance determined by using current NCCLS breakpoints, there was essentially no difference between azithromycin and clarithromycin, i.e., 0.5 and 1.9%, respectively (P = 0.086) . Interpretive breakpoints for erythromycin MIC tests versus H . influenzae have not been developed . Resistance to other non- beta-lactam agents was variable, as follows: trimethoprim-sulfamethoxazole, 9.0%; chloramphenicol, 0.2%; tetracycline, 1.3%; and rifampin, 0.3% . Two conspicuous findings in this study were the identification of 39 strains H . influenzae that were beta-lactamase negative but ampicillin intermediate or resistant (BLNAR) and, even more surprisingly, 17 beta-lactamase-positive isolates that were resistant to amoxicillin-clavulanate (BLPACR) . Strains of H . influenzae in the first group have heretofore been very uncommon; organisms in the second group have not previously been described in the literature . The percentages of all study isolates comprised of BLNAR and BLPACR organisms were 2.5 and 1.1, respectively . Overall resistance to ampicillin was thus 38.9%, and that to amoxicillin-clavulanate was 4.5%.

Soc Sci Med, 1997 Feb, 44(4), 441 - 54
STDS in women attending family planning clinics: a case study in Addis Ababa; Duncan ME et al.; For cultural reasons modern contraception has been slow to gain acceptance in Ethiopia . Knowledge about contraception and abortion is still limited in many family and community settings in which it is socially disapproved . By 1990 only 4% of Ethiopian females aged 15-49 used contraception . Little is known of sexually transmitted disease (STD) prevalence in family planning (FP) attenders in Africa in general and Ethiopia in particular, even though attenders of family planning clinics (FPCs) are appropriate target groups for epidemiological studies and control programmes . A study of 2111 women of whom 542 (25.7%) attended FPCs in Addis Ababa showed utilisation rates to be highest in women who were: Tigre (33%) or Amhara (31%), aged 20-34 years (30%), age 16 or older at first marriage/coitus (28%:38% in those first married after 25 years); who had a monthly family income of 10 Ethiopian Birr (EB) or more (33%:36% for those with income 100-500 EB), three or more children (37%), more than five lifetime husbands/sexual partners (39%); or were bargirls (73%) or prostitutes (43%) . The seroprevalence rates for all STDs, higher in FPC attenders compared with other women, were syphilis (TPHA) 39%, Neisseria gonorrhoeae 66%, genital chlamydia 64%, HSV-2 41%, HBV 40% and Haemophilus ducreyi 20% . Only 4% of FPC attenders had no serological evidence of STD: 64% were seropositive for 3 or more different STD . Clinical evidence of pelvic inflammatory disease (PID) was also more common in the FPC attenders (54%), 37% having evidence of salpingitis . The FPC provides a favourable setting for screening women likely to have high seroprevalence of STD, who for lack of symptoms will not attend either an STD clinic nor a hospital for routine check up . We recommend that measures be taken to adequately screen, treat and educate FPC attenders, their partners, and as appropriate and when possible their clients, in an attempt to control STDs and ultimately HIV in the community . Social, economic and cultural factors in the occurrence of STDs, prostitution, family planning and modern contraception coverage in Ethiopia are identified and deficiencies of current programmes briefly discussed with the objective of targeting services more effectively.

Infect Immun, 1997 Feb, 65(2), 651 - 60
Identification of tandem genes involved in lipooligosaccharide expression by Haemophilus ducreyi; Stevens MK et al.; A transposon insertion mutant of Haemophilus ducreyi 35000 possessing a truncated lipooligosaccharide (LOS) failed to bind the LOS-specific monoclonal antibody 3E6 (M . K . Stevens, L . D . Cope, J . D . Radolf, and E . J . Hansen, Infect . Immun . 63:2976-2982, 1995) . This transposon was found to have inserted into the first of two tandem genes and also caused a deletion of chromosomal DNA upstream of this gene . These two genes, designated lbgA and lbgB, encoded predicted proteins with molecular masses of 25,788 and 40,236 Da which showed homology with proteins which function in lipopolysaccharide biosynthetic in other gram-negative bacteria . The tandem arrangement of the lbgA and lbgB genes was found to be conserved among H . ducreyi strains . Isogenic LOS mutants, constructed by the insertion of a cat cartridge into either the lbgA or the lbgB gene, expressed an LOS phenotype indistinguishable from that of the original transposon-derived LOS mutant . The wild-type LOS phenotype could be restored by complementation with the appropriate wild-type allele . These two LOS mutants proved to be as virulent as the wild-type parent strain in an animal model . A double mutant with a deletion of the lbgA and lbgB genes yielded equivocal results when its virulence was tested in an animal model.

J Bacteriol, 1997 Feb, 179(3), 805 - 12
Neisseria meningitidis tonB, exbB, and exbD genes: Ton-dependent utilization of protein-bound iron in Neisseriae; Stojiljkovic I et al.; We have recently cloned and characterized the hemoglobin (Hb) receptor gene, hmbR, from Neisseria meningitidis . To identify additional proteins that are involved in Hb utilization, the N . meningitidis Hb utilization system was reconstituted in Escherichia coli . Five cosmids from N . meningitidis DNA library enabled a heme-requiring (hemA), HmbR-expressing mutant of E . coli to use Hb as both porphyrin and iron source . Nucleotide sequence analysis of DNA fragments subcloned from the Hb-complementing cosmids identified four open reading frames, three of them homologous to Pseudomonas putida, E . coli, and Haemophilus influenzae exbB, exbD, and tonB genes . The N . meningitidis TonB protein is 28.8 to 33.6% identical to other gram-negative TonB proteins, while the N . meningitidis ExbD protein shares between 23.3 and 34.3% identical amino acids with other ExbD and TolR proteins . The N . meningitidis ExbB protein was 24.7 to 36.1% homologous with other gram-negative ExbB and TolQ proteins . Complementation studies indicated that the neisserial Ton system cannot interact with the E . coli FhuA TonB-dependent outer membrane receptor . The N . meningitidis tonB mutant was unable to use Hb, Hb-haptoglobin complexes, transferrin, and lactoferrin as iron sources . Insertion of an antibiotic cassette in the 3' end of the exbD gene produced a leaky phenotype . Efficient usage of heme by N . meningitidis tonB and exbD mutants suggests the existence of a Ton-independent heme utilization mechanism . E . coli complementation studies and the analysis of N . meningitidis hmbR and hpu mutants suggested the existence of another Hb utilization mechanism in this organism.

J Cell Biol, 1997 Jan 27, 136(2), 271 - 85
A novel membrane-associated metalloprotease, Ste24p, is required for the first step of NH2-terminal processing of the yeast a-factor precursor; Fujimura-Kamada K et al.; Many secreted bioactive signaling molecules, including the yeast mating pheromones a-factor and alpha-factor, are initially synthesized as precursors requiring multiple intracellular processing enzymes to generate their mature forms . To identify new gene products involved in the biogenesis of a-factor in Saccharomyces cerevisiae, we carried out a screen for MA Ta-specific, mating-defective mutants . We have identified a new mutant, ste24, in addition to previously known sterile mutants . During its biogenesis in a wild-type strain, the a-factor precursor undergoes a series of COOH-terminal CAAX modifications, two sequential NH2-terminal cleavage events, and export from the cell . Identification of the a-factor biosynthetic intermediate that accumulates in the ste24 mutant revealed that STE24 is required for the first NH2-terminal proteolytic processing event within the a-factor precursor, which takes place after COOH-terminal CAAX modification is complete . The STE24 gene product contains multiple predicted membrane spans, a zinc metalloprotease motif (HEXXH), and a COOH-terminal ER retrieval signal (KKXX) . The HEXXH protease motif is critical for STE24 activity, since STE24 fails to function when conserved residues within this motif are mutated . The identification of Ste24p homologues in a diverse group of organisms, including Escherichia coli, Schizosaccharomyces pombe, Haemophilus influenzae, and Homo sapiens, indicates that Ste24p has been highly conserved throughout evolution . Ste24p and the proteins related to it define a new subfamily of proteins that are likely to function as intracellular, membrane-associated zinc metalloproteases.

MMWR Morb Mortal Wkly Rep, 1997 Jan 24, 46(3), 62 - 4
Antibiotic resistance among nasopharyngeal isolates of Streptococcus pneumoniae and Haemophilus influenzae--Bangui, Central African Republic, 1995; Synthesis of L-glycero-D-manno-heptopyranose-containing oligosaccharide structures found in lipopolysaccharides from Haemophilus influenzae; Department of Organic Chemistry, Arrhenius Laboratory, University of Stockholm, SwedenSyntheses are described of the tetrasaccharide 2-(4-trifluoroacetamidophenyl)ethyl O-(beta-D-galactopyranosyl)-(1-->2)-O- (L-glycero-alpha-D-manno-heptopyranosyl)-(1-->2)-O-(L-glycero-alpha-D- manno-heptopyranosyl)-(1-->3)-L-glycero-alpha-D-manno-heptopyranoside (20) and the three trisaccharides 2-(4-trifluoroacetamidophenyl)ethyl O-(L-glycero-alpha-D-manno-heptopyranosyl)-(1-->2)-O-(L-glycero-alpha-D- manno-heptopyranosyl)-(1-->3)-L-glycero-alpha-D-manno-heptopyranoside (17), 2-(4-trifluoroacetamidophenyl)ethyl O-(beta-D-glucopyranosyl)-(1-->4)- O-(beta-D-glucopyranosyl)-(1-->4)-L-glycero-alpha-D-manno-heptopyrano side (5), and 2-(4-trifluoro-acetamidophenyl)ethyl O-(beta-D-galactopyranosyl)-(1-->4)-O-(beta-D-glucopyranosyl)-(1-->4)- L-glycero-alpha-D-manno-heptopyranoside (8), corresponding to structures found in the lipooligosaccharides of Haemophilus influenzae.

MMWR Morb Mortal Wkly Rep, 1997 Jan 17, 46(2), 35 - 40
Recommended childhood immunization schedule--United States, 1997; Bacterial findings in acute maxillary sinusitis--European study; Department of Otorhinolaryngology, Tampere University Hospital, FinlandBacteriology of acute maxillary sinusitis was studied in 569 patients in 16 centers of 6 countries located throughout Europe during 1992-1994 by ENT specialists . Patients with symptoms of acute sinusitis lasting less than 3 weeks with ongoing purulent nasal discharge were included . Diagnosis was verified by sinus x-ray or ultrasonography and a positive aspiration finding in maxillary sinus puncture . One or more pathogens were isolated from the maxillary sinus aspirates of 375 (66%) patients . Fifty-six percent of patients harboured 1 pathogen and 10% multiple pathogenic organisms, respectively . Haemophilus influenzae was the most common pathogen isolated (148 isolates), occurring as a single pathogen in 14% of the patients . The occurrence of H . influenzae was highest in Finnish military hospital patients (43-48%), as compared with the non-military Finnish patients (9-11%) or to patients from other European centers (mean 13%) . H . influenzae was more frequently beta-lactamase positive in other European centers (22%) than in Finnish centers (7%) . Streptococcus pneumoniae was the most common pathogen isolated in other European centers (20%) but second most common in Finnish centers (13%) . Moraxella catarrhalis occurred at quite similar frequency among Finnish centers (9-14%), but clearly less often in other centers (mean 4%) . S . aureus, which in acute maxillary sinusitis is regarded as a contaminant from the nasal cavity, was more prevalent in other European centers (12%) than in Finnish centers (4%) . In patients with acute maxillary sinusitis reliable bacteriological samples should be taken by antral aspiration directly from the diseased sinus.

Acta Otolaryngol Suppl, 1997, 529, 162 - 4
Bacteriological evaluation of 194 adult patients with acute frontal sinusitis and findings of simultaneous maxillary sinusitis; Antila J et al.; In a prospective study bacteriological findings from frontal and maxillary sinus secretes were analyzed . Frontal sinus trephination was performed to 103 and maxillary irrigation to 192 patients . Main pathogens in the frontal as well as in the maxillary sinus secretes were Haemophilus influenzae and Streptococcus pneumoniae . beta-lactamase production of H . influenzae was found in 17% of frontal and in 16% of maxillary sinus samples . Bacteriological findings were supported by the staining results . The portion of anaerobic pathogens in cultures were poor, though Bacteroides fragilis antigen was detected in 8 frontal and 6 maxillary sinus samples . The acute frontal sinusitis should be treated with respective antibiotics . Complications seem not to develop in the acute phase and the disease can primarily be treated conservatively . It should however be controlled properly to avoid prolonged disease and late complications.

Acta Otolaryngol Suppl, 1997, 529, 144 - 7
Do simple laboratory tests help in etiologic diagnosis in acute maxillary sinusitis?
Savolainen S, Jousimies-Somer H, Karjalainen J, Ylikoski J.
The aim of the present study was to evaluate whether the results of such simple hematologic tests as erythrocyte sedimentation rate (ESR), white blood cell count (WBC) and C-reactive protein (CRP) could give any useful information about the causative agents in 176 patients with acute maxillary sinusitis (AMS) . The great majority of tests (82%) showed values which were within normal limits . This may be due to the fact that 22% of AMS cases were culture negative for bacteria and about 60% of culture positive cases had Haemophilus influenzae as the etiologic agent . Significantly raised test values were seen in connection with Streptococcus pyogenes in the majority of cases, less frequently with Streptococcus pneumoniae and rarely with Haemophilus influenzae . We conclude that none of the evaluated routine blood tests are particularly sensitive indicators of the specific etiology of AMS in general . However, elevated CRP values (> 40 mg/l) associated with AMS should alert the physician to the suspicion of Streptococcus pyogenes or Streptococcus pneumoniae in etiology, since both, if left untreated, may lead to sinus empyema . With CRP or other evaluated tests Haemophilus influenzae- or Branhamella catarrhalis-positive AMS cannot be distinguished from a purely viral disease.

Acta Otolaryngol Suppl, 1997, 529, 19 - 21
Are there microbiological markers to predict recurrent acute otitis media?
Rynnel-Dagoo B.
The nasopharynx is a natural reservoir for several bacterial species, including Streptococcus pneumoniae and Haemophilus influenzae . Carriership is a potential mechanism for pathogenicity, since bacteria might invade the Eustachian tube and cause middle ear disease . Whether the pattern of nasopharyngeal colonization is different in infection prone vs healthy children is still a matter of controversy . In several studies it has been shown that H . influenzae is carried significantly more often in otitis-prone children compared with healthy control children . Colonization with H . influenzae in young children may be regarded as a candidate for a microbiological marker for recurrent episodes of acute otitis media.

SAAS Bull Biochem Biotechnol, 1997, 10, 49 - 58
Structure of the Haemophilus influenzae uvr-1+ gene: homology with other uvrC-like genes and characterization of the Haemophilus influenzae uvr-1 and uvr-2 mutations; Gottschalk VA et al.; A 3.9 kb Haemophilus influenzae genomic DNA fragment was cloned in plasmid pUC9 that partially complemented the ultraviolet light sensitivity (UVs) of Escherichia coli uvrC mutant hosts . This fragment also complemented the UVs of H . influenzae uvr-1 (DB112) and uvr-2 (DB116) mutants . It genetically transformed the latter mutants to wild type UV resistance . The nucleotide (nt) sequence of this fragment revealed 3 open reading frames (ORFs) . ORF2, now designated uvr-1+ (1746 nt), shows significant similarity in both the nt and amino acid (aa) composition to 7 complete proven or putative uvrC gene sequences . Computer analysis of the DNA sequence revealed several possible regulatory motifs 5' to uvr-1+, including a putative LexA-binding site as an inverted SOS box, located within the 3' region of ORF1, (extensive homology to the E . coli CMP-KDO synthetase gene), upstream of uvr-1+ . Further computer analysis has also predicted that the four putative active site amino acids, located in the C-terminal half of each protein, are each situated in an area of secondary structure that are highly conserved.

Dev Biol Stand, 1997, 89, 379 - 89
Are serological responses to acellular pertussis antigens sufficient criteria to ensure that new combination vaccines are effective for prevention of disease?
Granoff DM, Rappuoli R.
Combining several vaccines in a single formulation can alleviate the increasing complexity of the paediatric vaccination schedule . However, vaccine antigens that are highly effective when administered singly may lose potency in combination; consequently, the efficacy of each component must be established for any new proposed combination vaccine . When the serological response to a vaccine correlates clearly with clinical efficacy, the efficacy of that component in a combination can be inferred from immunogenicity studies . Poliovirus, tetanus and diphtheria toxoids, Haemophilus influenzae type b and Neisseria meningitidis, and hepatitis B vaccines, can all be assessed in this manner . Unfortunately, the antibody titres induced by acellular pertussis vaccines do not correlate with vaccine efficacy . Thus, although diphtheria-tetanus-acellular pertussis (DTaP) vaccine has been considered a prime building block in the development of new combination vaccines, modifying DTaP by the addition of new vaccine components may decrease the ability of the vaccine to protect against pertussis without a change in serum antibody response . For this reason, immunogenicity is not an adequate or safe basis for licensing combination vaccines containing acellular pertussis . Development and licensing of new combination vaccines containing components with serological correlates of clinical efficacy can proceed more rapidly than DTaP-based combinations and should be pursued.

Indian Pediatr, 1997 Jan, 34(1), 9 - 15
Haemophilus influenzae type b vaccine in India: need and timing, immunogenecity and tolerance; Acharya D et al.; OBJECTIVE: (i) To assess the natural immunity and susceptibility to Haemophilus influenzae type b (Hib) infections in children in India . (ii) To study the immunogenecity and tolerance of Hib vaccine (ACTHIB) in young infants . DESIGNS: (i) Cross sectional study . (ii) Prospective trial . SETTING: Well baby and immunization clinics . METHODS: (i) PRP antibody titers against Hib estimated in 172 healthy infants and children aged 1 month to 10 years . (ii) Antibody titres estimated before and after ACTHIB vaccine given with primary immunization (age group 6 to 8 weeks) in 50 babies . RESULTS: (i) Naturally protective levels of Hib antibodies found in less than 20% of infants under one year, but in over 80% above 4 years . (ii) Seroconversion after ACTHIB vaccination was 100% with very high protective levels . There were no significant adverse reactions . CONCLUSIONS: ACTHIB vaccine proved to be safe and highly immunogenic . As susceptibility to Hib is highest in the first year of life, the vaccine should be recommended in the primary immunization schedule (combined with DPT) . The very high titers achieved suggest the possibility of decreasing the number of doses or the amount of antigen to reduce the prevalent high cost.

Ciba Found Symp, 1997, 207, 223 - 31; discussion 231-7
Epidemiological factors influencing the emergence of antimicrobial resistance; Cohen ML; Antimicrobial resistance is becoming an important public health problem for both hospital- and community-acquired infections . In the hospital, infections caused by drug-resistant Staphylococcus aureus, Mycobacterium tuberculosis, enterococci, and a variety of Gram-negative rods are resulting in increased morbidity, mortality and costs, in part because of prolonged hospitalization and the use of more expensive antimicrobial agents . Drug-resistant, community-acquired infections are also causing important problems in both the developed and the developing world . Although the relative importance of specific pathogens varies with the geographical area, community-acquired pathogens including Salmonella, Shigella, Neisseria gonorrhoeae, Haemophilus influenzae and Streptococcus pneumoniae are causing both sporadic cases and outbreaks of drug-resistant illness . The emergence of antimicrobial resistance is being attributed to a series of societal, technological, environmental and microbial changes . These include increasing populations of susceptible hosts, international travel and commerce, changes in technology and industry, microbial adaptation and change, and the breakdown of public health measures . Addressing emerging problems and antimicrobial resistance will require enhanced surveillance, prudent use of existing antimicrobial drugs, development of new antimicrobial agents, increased emphasis on infection control and hygienic practices, effective disease control programs, better use of existing vaccines, and development of more and better vaccines.

Fundam Clin Pharmacol, 1997, 11(1), 90 - 5
Pharmacokinetics and pharmacodynamics of two oral forms of cefuroxime axetil; Garraffo R et al.; Cefuroxime axetil is a cefuroxime ester that can be administered by mouth . Two dosage forms (tablets and granules) have been developed for oral administration . We evaluated the pharmacokinetics and pharmacodynamics of these forms in an open cross-over study involving 12 healthy volunteers receiving single doses of 250 mg . The bioavailability of the two forms was different, the observed peak concentration and time-concentration curve values of the tablet form being, respectively, 39 and 27% higher than those of the granule form . However, ex vivo studies of serum bactericidal activity against Streptococcus pneumoniae showed no significant differences between the two formulations . This is in keeping with the fact that the bactericidal activity of samples from only six subjects gave evaluable data for Haemophilus influenzae; although small differences were found between the two formulations, further investigations are required . The pharmacodynamic approach is becoming an essential element in determining the equivalence of antibiotic dosage forms.

Respiration, 1997, 64(3), 224 - 8
Hospitalized patients with community-acquired pneumonia in Hong Kong: a randomized study comparing imipenem/cilastatin and ceftazidime; Ho A et al.; The aetiology and outcome of hospitalized patients with moderate to severe community-acquired pneumonia (CAP) were evaluated in 60 adult patients (38 male 22 female, mean age 68.4 years) . They were randomized for treatment with either ceftazidime or imipenem/cilastatin intravenously for 7 days . Bacteriological diagnoses were made in 25 cases (41.6%): Streptococcus pneumoniae (5), Haemophilus influenzae (5), Pseudomonas spp . in particular Pseudomonas aeruginosa (8), Staphylococcus aureus (4), Chlamydia spp . (2), Mycobacterium tuberculosis (2) and Moraxella catarrhalis (3); mixed organisms were found in 4 patients . Forty-two patients (70%) responded satisfactorily to the regimens with improvement in sputum purulence cough and dyspnoea scores; there was no difference in response between the two groups . Sixteen patients (26.6%) underwent bronchoscopy on day 4 because of inadequate response to the antibiotics regimens, and 9 of them (15%) required a modification of the initial treatment with addition of erythromycin in 5 patients vancomycin in 1 cloxacillin in 1 and antituberculous drugs in 2 . Three out of the 60 patients (5%) died of pulmonary sepsis: the aetiological agents were M . tuberculosis in one, Pseudomonas spp./methicillin-resistant S . aureus in another, but were not identified in the third . We conclude that treatment with either ceftazidime or imipenem/cilastatin was efficacious for moderate to severe CAP in Hong Kong.

Enferm Infecc Microbiol Clin, 1997 Jan, 15(1), 5 - 9
{Comparative study of the protective role of the vaccine against Haemophilus influenzae type b and partial splenectomy in an experimental model}; Perez Maestu R et al.; BACKGROUND: Although the vaccine against Haemophilus influenzae type b has demonstrated its efficacy in children, this bacteria continues to be a cause or overwhelming sepsis in splenetomized patients . METHODS: To compare the degree of protection provided by partial splenectomy and the conjugated diptheria toxoid vaccine against H . influenzae type b, the bacteremia generated 24, 48 and 96 hours after intravenous inoculation with this bacteria at concentrations of 5 x 10(14), 5 x 10(13) and 5 x 10(12) colony-forming units was assessed in rats subjected to total splenectomy, with and without previous vaccination, to partial splenectomy or to sham operation . RESULTS: With respect to both the proportion of positive blood cultures and the concentration of H . influenzae in the blood stream, the sham-operated control rats and those vaccinated prior to splenectomy presented similar behaviors; the widest differences with respect to the latter two groups were found in the nonvaccinated total splenectomy rats, and the results with the partial splenectomy group fell between these two extremes . CONCLUSIONS: The protection of H . influenzae type b vaccine against infection is equivalent to that provided by the intact spleen for inoculum concentrations of 5 x 10(13) colony-forming units and less . Thus, the possibility of its systematic use in asplenic patients should be considered . The protection provided by the remnant spleen following 50% splenectomy is intermediate between those observed with eusplenia and with asplenia . These results suggest that when partial splenectomy is to be performed in normal spleen, the attempt should be made to conserve as much of the organ as possible.

Diagn Microbiol Infect Dis, 1997 Jan-Feb, 27(1-2), 35 - 9
The antibiotic treatment of acute otitis media and sinusitis in children; Cohen R; The development of resistance among the bacterial pathogens causing acute otitis media and sinusitis in children is causing considerable concern . Although normally a mild infection, acute otitis media can produce serious complications with sequelae that can have long-lasting effects . High levels of resistance are now being seen in the three principal pathogens . Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis . Most clinical trials comparing the efficacy of different antibiotics have failed to show differences in clinical efficacy . To overcome this problem, alternative methods of assessing antibiotics have been developed including the "in-vivo sensitivity test" proposed by Howie and retrospective analyses of treatment failures . The treatment of sinusitis is complicated by the difficulty in establishing a clear differential diagnosis . To date, no definitive trials of bacteriologic efficacy in children have been published.

Diagn Microbiol Infect Dis, 1997 Jan-Feb, 27(1-2), 21 - 8
Can antimicrobial activity be sustained? An appraisal of orally administered drugs used for respiratory tract infections; Jones RN; Respiratory tract infections (RTIs) represent a major cause of illness worldwide . Therefore, it is of great concern that common RTI pathogens have become increasingly resistant to many of the antimicrobial agents used for therapy . For example, Haemophilus influenzae and Moraxella catarrhalis have become resistant to beta-lactam drugs by producing efficient beta-lactamases (> 35 and 90% of strains, respectively) . More recently, pneumococci have become more resistant through the mechanism of altered penicillin-binding proteins (PBPs) . The rate of penicillin nonsusceptible isolates has risen to > 25% in the United States (1994-1995) . It is important to monitor the resistance characteristics of such pathogens and, if possible, to use regionally acquired data to guide empiric selection of therapeutic agents for RTIs . Currently, some antimicrobials remain effective against the majority of these three bacterial species, as exemplified by amoxicillin/clavulanic acid . Furthermore, amoxicillin alone seems to possess greater inhibition than other orally administered beta-lactams at clinically achievable concentrations against pneumococci with altered PBPs . It is critical that steps are taken to limit resistance problems, particularly through; 1) education of prescribers and the public; 2) initiation of the development of novel drugs with alternative modes of action or stability to existing resistance mechanisms; and 3) by continuing to generate quality susceptibility testing data to guide empiric chemotherapy against bacterial pathogens causing RTI.

J Infect, 1997 Jan, 34(1), 69 - 74
HACEK group endocarditis at the Riyadh Armed Forces Hospital; el Khizzi N et al.; INTRODUCTION: Fastidious Gram-negative organisms classified as the HACEK group (Haemophilus spp., Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens and Kingella spp.) are rare causes of infective endocarditis . CASE REPORT: In this series, we report six cases of endocarditis in Saudi patients occurring between 1990 and 1994 in our hospital, caused by two of the organisms in the HACEK group, i.e . Cardiobacterium hominis and Actinobacillus actinomycetemcomitans . The clinical features, predisposing factors and treatment of the patients are briefly described . The microscopic and cultural characteristics of the organisms are described together with the laboratory methods of diagnosis . MAJOR FINDINGS: HACEK endocarditis was frequently associated with prosthetic heart valves or structural heart abnormalities . Dental caries or periodontal disease seems to be a predisposing factor . The prognosis of HACEK endocarditis is very good as clinical and bacteriological cure were achieved with antibiotic therapy in all our cases except one who required mitral valve replacement . Contrary to previous reports we did not find all the organisms sensitive to penicillin and aminoglycosides . However, all our isolates were sensitive to amoxycillin, cefuroxime, ceftriaxone and ciprofloxacin . CONCLUSIONS: Laboratory diagnosis of HACEK group of organisms requires a high index of suspicion and should be suspected in cases of endocarditis in which fastidious Gram-negative coccobacilli are isolated which fail to grow on MacConkey agar . Empiric therapy should be started in suspected cases with second generation cephalosporins or with ciprofloxacin, until antibiotic sensitivity results become available . Antibiotic therapy should be continued for 4-6 weeks . Progress and outcome was very good in the series.

J Infect, 1997 Jan, 34(1), 41 - 8
The epidemiology of community-acquired pneumonia among hospitalized adults; Porath A et al.; OBJECTIVE: To identify and characterize the aetiological agents of community-acquired pneumonia (CAP) among hospitalized patients, as an aid in therapeutic decision-making . METHOD: A prospective 1 year study of all patients hospitalized with CAP in the Negev region of Israel . The aetiology was determined by blood and pleural fluid cultures, and specific serological testing for pathogenic agents . Eighty-nine percent of the patients underwent follow-up for a month after discharge . RESULTS: The study included 346 patients (53% males, mean age 49.3 +/- 19.5, range 17-94) . A single aetiologic agent was identified in 146 patients (42.2%), multiple agents were found in 133 (38.4%), and no agent was identified in 67 (19.4%) . Among the common pathogens were Pneumococcus sp . in 148 patients (42.8%) . Mycoplasma pneumoniae (101, 29.2%), Chlamydia pneumoniae (62, 17.9%), Legionella sp . (56, 16.2%), viruses (35, 10.1%), Coxiella burnetti (20, 5.8%) . Haemophilus influenzae (19, 5.5%), and other bacteria (21, 6.1%) . Approximately 70% of the patients infected with M . pneumoniae and C . burnetti were younger than 45 years (P < 0.05) . In contrast, about 50% of the patients with C . pneumoniae (TWAR) were over the age of 65 (P = 0.03) . The presence of comorbidity was associated with a greater frequency of bacterial aetiologies (57% vs . 44%, P = 0.02), and fewer infections with M . pneumoniae (15% vs . 36%, P = 0.0004), or C . burnetii (2% vs . 8%, P = 0.02) . Specific causative agents were associated with specific seasons: viruses between December and April (P = 0.03), and Legionella sp . from July to October (P = 0.003) . In contrast, no seasonal variation was associated with pneumococcus, M . pneumoniae, or C . pneumoniae (TWAR) . CONCLUSIONS: Patients are hospitalized with CAP throughout the year . Since the pathogen is usually unknown at hospitalization, epidemiological data is important for choosing medication . The findings of this study point to the importance of macrolides alone or in combination with cephalosporins, as the treatment of choice for patients in our region.

Pediatrics . 1997 Jan;99(1):E4.
Increasing immunization: a Medicaid managed care model; Browngoehl K et al.; OBJECTIVE:To evaluate the impact of an immunization outreach program on immunization rates . SETTING: A Pennsylvania independent practice association model managed care organization (100% Medicaid) . DESIGN: Retrospective cohort study (N = 2511) of children 30 to 35 months of age from two age cohorts that compared immunization rates for Advisory Committee on Immunization Practices schedules for diphtheria-tetanus-pertussis, oral polio vaccine, measles-mumps-rubella, and Haemophilus influenza type b . An evaluation of the outreach component of the program compared treatment and nontreatment subgroups of one age cohort (N = 1002) . INTERVENTION: The immunization program targeted approximately 19 000 members from birth to 6 years of age . The program components included computerized tracking and reminders, member and provider education, provider incentives, member incentives, and home visiting outreach . RESULTS: Data indicate that the treatment group has higher completed immunization rates at 35 months of age than does the control group . Furthermore, data show that members with home visits have significantly higher completed immunization rates than do other members . The corresponding comparisons for age-appropriate immunizations by 24 months indicate a nonsignificant trend of increased rates . CONCLUSION: The data provide evidence supporting a correlation between comprehensive strategies (computerized tracking, member and provider education and incentives, and home visiting) and increased immunization rates . Those individuals who received home visits were more likely to complete an immunization series by 35 months of age than those who did not . However, within the Mercy Health Plan program, age-appropriate immunizations are not significantly affected by home-visiting outreach.

Arch Pediatr, 1997 Jan, 4(1), 52 - 60
{Role of corticoids in purulent meningitis in children: analysis of literature studies}; Jacqz-Aigrain E et al.; Steroid therapy, in combination with antibiotics for bacterial meningitis in paediatric patients remains controversial . Steroids, and primarily dexamethasone a very potent anti-inflammatory agent, regulate the liberation of various cytokines and inflammatory mediators such as prostaglandins, released during bacterial meningitis and leading to long term complications . Several clinical trials studying infants and children with bacterial meningitis due to Haemophilus influenzae have evaluated the beneficial effects of the administration of dexamethasone at the onset of antibiotherapy and demonstrated that dexamethasone reduced the risk of acquired sensorineural deafness (bilateral moderate or more severe hearing loss) and the incidence of neurological sequelae . Limited information is available for the other bacterial meningitis, although meningococcal meningitis will become more frequent with the use of effective anti-Haemophilus vaccines . In addition some Streptococcus pneumoniae are now resistant to third generation cephalosporins and the use of dexamethasone in that case may be at risk . Finally, no evidence is available for an effective role for dexamethasone in neonatal bacterial meningitis, although it is quite often administered in that age group.

Glycoconj J, 1997 Jan, 14(1), 13 - 7
Chemical synthesis of Haemophilus influenzae glycopeptide conjugates; Kandil AA et al.; A simple procedure for conjugating synthetic fragments of the capsular polysaccharide of Haemophilus influenzae type b, poly-3-beta-D-ribose-(1,1)-D-ribitol-5-phosphate (sPRP) to linear peptides is described . The procedure consists of (i) reacting the amino group of amino-heptyl sPRP with m-maleimidobenzoyl-N-hydroxysuccinimide (MBS) in phosphate buffer, pH 7.5; (ii) selectively coupling the MBS-modified sPRP to the sulfhydryl group of the cysteine residue of peptides containing functional T-helper cell epitope(s) . The glycopeptide conjugates were purified by gel filtration chromatography, biochemically characterized, and elicited protective level of anti-PRP antibody responses in rabbits.

S Afr Med J, 1997 Jan, 87(1), 51 - 6
Epidemiology of post-neonatal bacterial meningitis in Cape Town children; Hussey G et al.; Bacterial meningitis is a major cause of childhood morbidity and mortality in South Africa . However, comprehensive regional or national epidemiological data, essential for rational public health interventions, are lacking . The purpose of this 1-year prospective study, from 1 August 1991 to 31 July 1992, was to define the magnitude of the problem of childhood bacterial meningitis in Cape Town . The study group consisted of all children, aged > 1 month to < 14 years, who presented with proven bacterial meningitis at all the hospitals in the Cape Town metropolitan area . During the year 201 cases were identified: 101 (50.2%) were due to Neisseria meningitidis, 74 (36.8%) were due to Haemophilus influenzae and 26 (12.9%) were due to Streptococcus pneumoniae . The overall incidence rate (95% confidence interval) for children less than 14 years, 5 years and 1 year was 34 (30-40), 76 (65-88) and 257 (213-309) per 100,000 children, respectively . The rate was highest in black infants, 416 (316-545)/100,000 . This was 2 times greater than the rate in coloured infants and about 4.5 times greater than the rate in white infants . The median age of all the children was 10 months . The ages of children with haemophilus and pneumococcal meningitis were similar, 9 and 7.5 months respectively (P = 0.43), while children with meningococcal meningitis were significantly older (22 months) than the others (P < 0.01) . The overall case fatality rate was 5%, and 12.9% of survivors had significant neurological sequelae (disability) on discharge.

Eur J Epidemiol, 1997 Jan, 13(1), 73 - 7
Incidence of invasive Haemophilus influenzae type b disease in Italian children; Tozzi AE et al.; To estimate the incidence of Haemophilus influenzae type b (Hib) invasive disease in Italian infants we performed a prospective study in a cohort of newborns enrolled for a randomized trial on safety and efficacy of three pertussis vaccines and followed for onset of serious disease or pertussis . The overall cumulative incidence observed in 15,601 children was 51.3/100,000 for all invasive Hib infections and 38.4/100,000 for Hib meningitis, over 27 months of observation . The incidence density of all invasive Hib disease was 28.7/100,000 person-years, while meningitis occurred with an incidence of 21.5/100,000 person-years . Among the eight cases detected, six were meningitis, one sepsis, and one cellulitis . The child with sepsis died . The incidence and epidemiology of invasive Hib disease in Italy are comparable to those reported from other European countries . Cost-benefit analyses are needed for planning Italian vaccination policy.

J Clin Pathol, 1997 Jan, 50(1), 72 - 4
Endocarditis caused by Haemophilus parainfluenzae identified by 16S ribosomal RNA sequencing; Das I et al.; Haemophilus parainfluenzae, a human commensal, is an infrequent cause of serious disease . A case of endocarditis caused by this organism in a five year old boy with complex congenital heart disease is reported . The course of this disease was very aggressive, leading to heart failure, disseminated intravascular coagulation and multiorgan failure in spite of appropriate antibiotics and surgical intervention . The difficulties in the detection and identification of H parainfluenzae using conventional culture based technology, and the potential role of molecular techniques, are highlighted.

Retina, 1997, 17(1), 28 - 32
Intravenous gentamicin and ceftazidime in penetrating ocular trauma: a swine model; Schech JM et al.; PURPOSE: A swine model of ocular trauma was used to determine the extent of penetration of systematically administered gentamicin and ceftazidime to the vitreous cavity of traumatized and normal eyes . METHODS: Forty-six pigs received a scleral laceration to the right eye and then underwent surgical repair . Thirty-six animals received intravenous gentamicin, and 10 pigs were given ceftazidime . RESULTS: The level of gentamicin in the vitreous of traumatized and nontraumatized eyes did not achieve the minimum inhibitory concentrations (MIC) for Pseudomonas and Haemophilus species despite multiple and large intravenous doses . Ceftazidime concentrations in traumatized eyes were above the minimum inhibitory concentration for these organisms . CONCLUSIONS: Ceftazidime achieves an intravitreal concentration 33 times the minimum inhibitory concentration of Haemophilus species and twice that of Pseudomonas species in traumatized eyes after systemic administration, a finding that lends support to its use as a prophylactic agent in the management of penetrating ocular trauma . Gentamicin does not appear to enter the traumatized eye at appreciable levels after systemic administration, and, therefore, it is not recommended for use as a prophylactic agent in the management of penetrating ocular trauma.

Vaccine, 1997 Jan, 15(1), 20 - 4
Vaccination coverage of 2-year-old children and immunization practices--Canada, 1994; Duclos P; The purpose of the study was to assess vaccination coverage of 2-year-olds and estimate immunization practices in Canada using a postal survey with a panel of 862 households . RIM weighting was used to provide national estimates . 95% CIs were calculated using the binomial method . Completed questionnaires were received for 534 children born between February 1991 and January 1992 . Thirty nine percent of children had received all/most of their vaccinations at a public health clinic . Overall 5.6% parents indicated they had their child immunized because it was mandatory and 7.4% because of requirements for school, pre-school, nursery or day care attendance . A total of 78.7% of parents remembered receiving information on benefits of immunizations and 97% on the risks/side-effects . By their second birthday, 83.7% (95% CI 80.3-86.7) of 2-year-olds had received complete immunization against poliomyelitis; 97.2% (95% CI 95.4-98.4), at least one dose of a measles containing vaccine; 86.5% (95% CI 83.3-89.3), at least one dose of Haemophilus influenzae type b vaccine; and 81.6% (95% CI 78.1-84.8), at least, four doses of diphtheria, tetanus and pertussis vaccine.

Acta Otolaryngol, 1997 Jan, 117(1), 109 - 12
The effect of antibiotic treatment on the release of endotoxin during nontypable Haemophilus influenzae-induced otitis media in the chinchilla; Jauris-Heipke S et al.; The gram negative bacteria, nontypable Haemophilus influenzae (NTHi) was used to induce otitis media in a total of 18 chinchillas . Three days post-inoculation, three cohorts of 6 chinchillas each were treated daily for four days with either ceftriaxone, chloramphenicol, or diluent without antibiotics . Middle ear fluid (MEF) was obtained daily, assayed for endotoxin content by means of the chromogenic limulus amebocyte lysate assay, and concentration of the NTHi/mL MEF determined by standard plate count . The endotoxin concentration per mL MEF from both the antibiotic treated cohorts decreased during the observation period, but increased in the MEF of the untreated control group . The data indicate that, unlike the dramatic increase in endotoxin concentration, after antibiotic treatment in the cerebrospinal fluid (CSF) during experimental Haemophilus influenzae-induced meningitis, there is no demonstrable sustained release of endotoxin in the middle ear subsequent to antibiotic treatment during experimental otitis media.

Microb Pathog, 1997 Jan, 22(1), 39 - 46
Attachment of Haemophilus ducreyi to human foreskin fibroblasts involves LOS and fibronectin; Alfa MJ et al.; Haemophilus ducreyi is the causative agent of the genital ulcer disease Chancroid . Chancroid has been shown to increase the risk of heterosexual transmission of HIV . Little is known regarding the attachment or localization of this organism to human cells in either the dermal or epidermal layer . In this study the attachment of H . ducreyi to human foreskin fibroblast (HFF) cells was further characterized . Attachment was mediated by more than one mechanism . Proteinase K treatment but not trypsinization of H . ducreyi significantly reduced attachment suggesting protein involvement . In addition, purified lipooligosaccharide (LOS) was able to inhibit attachment in a dose dependent manner . It appeared that the organism binds to fibronectin in the extracellular matrix of HFF cells, since competition studies using fibronectin showed that it was able to significantly reduce attachment in a dose dependent manner whereas collagen did not . We hypothesize that the attachment of H . ducreyi involves both a protein mediator of attachment (likely pili) as well as LOS and that one or both of these bacterial components interacts with fibronectin in the extracellular matrix to mediate attachment to HFF cells.

Microb Pathog, 1997 Jan, 22(1), 13 - 21
Modulation of phagocytic function of bovine mononuclear phagocytes by Haemophilus somnus; Gomis SM et al.; The interactions between bovine mononuclear cells and Haemophilus somnus are known to be complex . To study this interaction, a flow cytometric assay was developed to assess the effect of H . somnus on phagocytosis of killed opsonized Staphylococcus aureus by bovine alveolar macrophages and blood monocytes . Using this in vitro system, it was found that log phase H . somnus significantly inhibited the phagocytosis of killed opsonized S . aureus by bovine alveolar macrophages obtained both from healthy calves and from cattle experimentally infected with H . somnus . However, killed log-phase H . somnus, in vitro passaged and stationary phase H . somnus had no effect on the phagocytic activity of these cells . In contrast to bovine alveolar macrophages, blood monocytes showed a significant increase in their phagocytic activity following in vitro exposure to either log or stationary phase H . somnus . Using a lypophilic, non-toxic fluorophore PKH2 to label live H . somnus, it was possible to simultaneously measure the uptake of both S . aureus and H . somnus . Stationary and log phase H . somnus were taken up by macrophages equally well, even though phagocytosis of S . aureus was inhibited by only log phase H . somnus . These results demonstrate the ability of H . somnus to modulate bovine mononuclear phagocytic function which might contribute towards the pathogenesis of bovine hemophilosis.

Microbiology, 1997 Jan, 143 ( Pt 1), 171 - 4
A Bacillus cereus member of the SNF2 family; Lindback T et al.; The complete sequence of a Bacillus cereus member of the SNF2 family of putative helicases showed conservation of all seven motifs typical of this family . Bcsnf predicted a protein of 1064 aa where the conserved SNF2 domain was located at the carboxy terminus, whereas the 633 amino-terminal aa showed no homology to any protein in the databases . A putative transcriptional start was identified by primer extension, indicating that Bcsnf is not a part of a larger operon . No phenotypical changes were observed after insertional inactivation of Bcsnf . The completely sequenced genomes of Mycoplasma genitalium and Haemophilus influenzae contain one ORF each with similarity to the SNF2 family: MG018 and HI0616, respectively . A phylogenetic tree of the SNF2 family showed that BcSNF and MG018 were most closely related, and appeared closer to the eukaryotic members of the SNF2 family than to the two other bacterial members of the family, HepA from Escherichia coli and HI0616.

Nucleic Acids Res, 1997 Jan 1, 25(1), 246 - 7
The translational signal database, TransTerm: more organisms, complete genomes; Dalphin ME et al.; TransTerm is a database of initiation and termination sequence contexts from more than 250 organisms listed in GenBank, including the four complete genomes:Haemophilus influenzae, Methanococcus jannaschii, Mycoplasma genitalium,and Saccharomyces cerevisiae . For the current release, more than 60 000 coding sequences were analysed . The tabulated data include initiation and termination contexts organised by species along with quantitative parameters about individual coding sequences (length, %GC, GC3, Nc and CAI) . There are also tables of initiation- and termination-region nucleotide-frequencies, codon usage tables and summaries of stop signal usage . TransTerm is available on the World Wide Web at: tml

Nucleic Acids Res, 1997 Jan 1, 25(1), 37 - 8
The metabolic pathway collection: an update; Selkov E et al.; The Metabolic Pathway Collection from EMP is an extraction of data from the larger Enzymes and Metabolic Pathways database (EMP) . This extraction has been made publicly available in the hope that others will find it useful for a variety of purposes . The original release in October 1995 contained 1814 distinct pathways . The current collection contains 2180 . Metabolic reconstructions for the first completely sequenced organisms-Haemophilus influenzae,Mycoplasma genitalium,Saccharomyces cerevisiaeandMethanococcus janaschii-are all included in the current release . All of the pathways in the collections are available as ASCII files in the form generated by the main curator, Evgeni Selkov . In addition, we are offering a more structured encoding of a subset of the collection; our initial release of this subcollection includes all of the pathways inMycoplasma genitalium, and we ultimately intend to offer the entire collection in this form as well.

Heart Lung, 1997 Jan-Feb, 26(1), 80 - 2
Haemophilus influenzae sepsis resulting from pneumonia; Marinella MA; Haemophilus influenzae is a pleomorphic gram-negative bacterium that causes a myriad of infections in both adults and children . The organism frequently causes respiratory infections in patients with obstructive lung disease but may on occasion cause invasive infections including pneumonia with bacteremia . We report the case of a patient with underlying lung disease and metastatic malignancy in whom sepsis related to pneumonia caused by H . influenzae developed.

J Mol Evol, 1997 Jan, 44(1), 66 - 73
Conserved clusters of functionally related genes in two bacterial genomes; Tamames J et al.; An approach for genome comparison, combining function classification of gene products and sequence comparison, is presented . The genomes of Haemophilus influenzae and Escherichia coli are analyzed, and all genes are classified into nine major functional classes, corresponding to important cellular processes . To study gene order relationships and genome organization in the two bacteria, we performed statistics on neighboring pairs of genes . To estimate the significance of the observations, a statistical model based on binomial distributions has been developed . Significant patterns of gene order are observed within, as well as between, the two bacterial genomes: Functionally related genes tend to be neighbors more often than do unrelated genes . Some of these groups represent well-known operons, but additional gene clusters are identified . These clusters correspond to genomic elements that have been conserved during bacterial evolution . In addition to nearest-neighbor relationships, the method is also useful to study the relative direction of transcription in genomes, which is also highly conserved between homologous gene pairs . This new approach combines the high-level description of molecular function with pair statistics that express genome organization . It is expected to complement traditional methods of sequence analysis in the study of genomic structure, function, and evolution.

Eur J Pediatr, 1997 Jan, 156(1), 18 - 24
Evidence for induction of polysaccharide specific B-cell-memory in the 1st year of life: plain Haemophilus influenzae type b-PRP (Hib) boosters children primed with a tetanus-conjugate Hib-DTPa-HBV combined vaccine; Zepp F et al.; The lack of an adequate immune response to the major polysaccharide of the Haemophilus influenzae type b (Hib) capsule (polyribosyl ribitol phosphate) (PRP) in very young infants (< 18 months) can be overcome by conjugating PRP to a T-cell dependent carrier protein . We studied whether administration of a tetanus-PRP conjugate vaccine reconstituted with a diphtheria-tetanus-acellular pertussis-hepatitis B (DTPa-HBV) vaccine as a three dose primary course at 3, 4 and 5 months of age induced PRP-specific immunological memory, by examining the anti-PRP response to a dose of unconjugated PRP given with the DTPa-HBV booster approximately 1 year later . The unconjugated PRP elicited protective anti-PRP antibody levels (> or = 0.15 microgram/ml) in all but 3 of the 369 vaccinees, including 13 infants who failed to demonstrate a measurable immune response after the primary course . In a sub-cohort of 54 subjects all had anti-PRP levels > or = 0.5 microgram/ml within 7-14 days of the booster showing a rapid anamnestic type response . Both primary and booster responses were predominantly IgGl indicating a T-cell dependent response . The DTPa-HBV components elicited protective anti-diphtheria, anti-tetanus and anti-HBs antibody levels in > or = 98.5% of vaccinees, and immune responses to each of the acellular pertussis vaccine components in 92.3%-97.3% of subjects . CONCLUSION: The tetanus-PRP conjugate vaccine not only elicited a good primary humoral response, but also induced immunological memory so that the infants were able to mount a large and rapid immune response to subsequent exposure to plain PRP, indicating that protection against circulating wild-type Hib had been generated . Successful induction of immunological memory occurred even when there was no measurable humoral anti-PRP response to the primary course . Tetanus-PRP conjugate vaccine can be used in combination with DTPa-HBV vaccine, when administered separately or as a single injection in the same syringe, in primary immunisation schedules at 3, 4 and 5 months of age.

Pediatr Infect Dis J, 1997 Jan, 16(1), 91 - 5; discussion 95-6, 123-6
Pathogenesis of bacterial bronchitis in cystic fibrosis; Smith A; OBJECTIVE: To describe the current understanding of the pathogenesis of infections in cystic fibrosis (CF) . SUMMARY: The key element in the pathogenesis of infections in CF is the abnormal mucin resulting from a defective chloride channel . Abnormal mucin and the lack of hydration of respiratory secretions entraps bacteria, permitting colonization and subsequent infection . "Normal" physiologic insults, such as microaspiration, and pollution evoke mucin secretion with bacteria-mucin aggregates causing infection of small airways . Microcolonies of pulmotropic bacteria (i.e . Haemophilus and Staphylococcus) in the trapped mucin cause mucosal injury which predisposes the patient to Pseudomonas aeruginosa infection . Eventually there is obstruction of medium-sized and small airways by inflammatory exudate and mucus . Ultimately these chronic endobronchial bacterial infections cause significant loss of pulmonary function with morbidity and a decrease in the life span of patients with CF . Although antibiotic therapy is beneficial for the management of bacterial respiratory tract infections in CF patients, gene therapy may provide the ultimate cure . CONCLUSION: New treatment strategies will emerge as the pathogenesis of cystic fibrosis is better elucidated.

J Infect Dis, 1997 Jan, 175(1), 205 - 9
New murine model of bronchopneumonia due to cell-bound Haemophilus influenzae; Miyazaki S et al.; This murine model of nontypeable (unencapsulated) Haemophilus influenzae (NTHI) bronchopneumonia used organisms bound to mouse fetal lung (MFL) cells as an inoculum . Pretreatment of the mice with 40 microL of 1% formalin 3 days before intranasal instillation of the bacteria was necessary to allow infection . The number of NTHI recovered from the lungs plus trachea on day 7 after instillation was >100 times the number originally inoculated . Later, however, the number of recovered bacteria diminished gradually, and by day 14 it was almost identical to the original inoculum size . Serum IgM also peaked on day 7 after infection, after which IgG increased while IgM decreased . Histologically, bronchoalveolar infiltration of neutrophils was observed on day 3 after inoculation and continued at least for the following 4 days . The present experiment demonstrates that MFL cells can protect bacteria that have invaded the cells from the opsonizing and killing activities of host humoral defense mechanisms.

J Infect Dis, 1997 Jan, 175(1), 77 - 83
Receptor affinity purification of a lipid-binding adhesin from Haemophilus influenzae; Busse J et al.; Thirteen clinical strains of Haemophilus influenzae, including types b, d, and untypeable, in vitro specifically recognize phosphatidylethanolamine (PE), gangliotetraosylceramide, gangliotriosylceramide (Gg3), sulfatoxygalactosylceramide, and to a lesser extent sulfatoxygalactosylglycerol . A PE affinity matrix was used to purify an adhesin of approximately 46 kDa from both type b and untypeable H . influenzae . This adhesin was a potent inhibitor of H . influenzae Gg3 and PE binding in vitro, and polyclonal antibodies specific for this protein prevented the attachment of H . influenzae Gg3 and PE and cultured HEp-2 epithelial cells in vitro.

Infect Immun, 1997 Jan, 65(1), 261 - 6
Human Fab fragments specific for the Haemophilus influenzae b polysaccharide isolated from a bacteriophage combinatorial library use variable region gene combinations and express an idiotype that mirrors in vivo expression; Reason DC et al.; To determine whether the human antibody (Ab) repertoire to the Haemophilus influenzae type b capsular polysaccharide (Hib PS) could be studied at the molecular level with phage display technology, we constructed a phage Fab library by using peripheral blood from a vaccinated adult . Phage were selected based on Hib PS binding . Two distinct Hib PS-specific phage clones were identified whose Fab fragments used the same V(H) region paired with two different V(L) regions . The V(L) regions were derived from two independent rearrangements of the A2c gene with Jkappa1, and both contained a nontemplated arginine codon at the V-Jkappa junction . The two A2 V gene segments differed from the A2c germ line sequence in 0 and 5 bases . The V(H) region consisted of the V(H)26 gene segment having 98% identity to the germline nucleotide sequence, a D region of 9 bases, and J(H)4b1 . Usage of V(H)26 in combination with A2 V regions containing a junctional arginine is a predominant configuration of naturally occurring Hib PS-specific Abs . Liquid- and solid-phase assays showed that phage-derived Fab reacted with Hib PS and expressed HibId-1, an idiotype associated with the kappaII-A2 V region . These findings extend the database of V region polymorphisms that can contribute to the Hib PS repertoire and demonstrate that Hib PS-specific Fab fragments isolated from combinatorial phage libraries use V gene combinations which mirror the natural repertoire.

Infect Immun, 1997 Jan, 65(1), 150 - 5
Importance of an immunodominant surface-exposed loop on outer membrane protein P2 of nontypeable Haemophilus influenzae; Yi K et al.; Nontypeable Haemophilus influenzae (NTHI) frequently causes recurrent infections of the respiratory tract in humans . Previous indirect evidence suggested that a strain-specific immune response occurs following infection and that this immune response is directed at an immunodominant epitope on the bacterial surface . To test this hypothesis, mice and rabbits were immunized with whole cells of a strain of NTHI and the antiserum was characterized to identify the antigens to which antibodies were directed . All animals made a prominent antibody response to the loop 5 region of the P2 molecule, which is the major outer membrane protein . Rabbit serum showed complement-dependent bactericidal activity . Adsorption of the immune serum with the loop 5 fusion peptide removed bactericidal activity and also abolished reactivity to P2 detected by an immunoblot assay, an enzyme-linked immunosorbent assay, and a radioimmunoprecipitation assay . These data indicate that immunization with whole cells of NTHI results in a prominent antibody response which is directed at epitopes on the loop 5 region of the P2 molecule . Thus, a strain-specific immune response to NTHI occurs as a result of the expression of an immunodominant epitope on the P2 molecule.

J Clin Microbiol, 1997 Jan, 35(1), 125 - 31
Interpretive criteria and quality control parameters for testing of susceptibilities of Haemophilus influenzae and Streptococcus pneumoniae to trimethoprim and trimethoprim-sulfamethoxazole . The Antimicrobial Susceptibility Testing OC Group; Fuchs PC et al.; Two hundred twenty-eight strains of Haemophilus influenzae and 234 strains of Streptococcus pneumoniae were tested by broth microdilution and disk diffusion methods for susceptibility to trimethoprim (TMP) and TMP-sulfamethoxazole (SMX) to evaluate proposed criteria . Data are presented to support the proposed TMP MIC breakpoints of < or = 2.0 micrograms/ml for susceptibility and > or = 4.0 micrograms/ml for resistance for both species and TMP-SMX MIC breakpoints of < or = 2.0-38 micrograms/ml for susceptibility and > or = 4.0-76 micrograms/ml for resistance . Corresponding zone diameter breakpoints for H . influenzae for both drugs are proposed: < or = 10 mm = resistant; > or = 16 mm = susceptible . A 10-laboratory study documented reproducibility of such tests with standard control strains . The following control limits are proposed for tests of H . influenzae ATCC 49247 against TMP; MIC, 0.12 to 0.5 microgram/ml; zone diameter, 27 to 33 mm . The current limits for TMP-SMX were confirmed . For tests of S . pneumoniae ATCC 49619, MICs of TMP were 1.0 to 4.0 micrograms/ml and the current TMP-SMX MIC range was confirmed . Disk susceptibility tests of either drug against pneumococci were not reproducible, and consequently neither quality control limits nor interpretive criteria could be established . Endpoint interpretation and lot-to-lot variability in Mueller-Hinton agars were significant factors leading to interlaboratory variability.

FEMS Immunol Med Microbiol, 1996 Dec 31, 16(3-4), 173 - 81
Characterization of monoclonal antibodies that recognize common epitopes located on O antigen of lipopolysaccharide of serotypes 1, 9 and 11 of Actinobacillus pleuropneumoniae; Rodriguez Barbosa JI et al.; Seven murine monoclonal antibodies (mAbs) against serotype 1 of Actinobacillus (Haemophilus) pleuropneumoniae (reference strain Shope 4074) were produced and characterized . All hybridomas secreting mAbs were reactive with whole-cell antigens from reference strains of serotypes 1, 9 and 11, except for mAb 5D6 that failed to recognized serotype 9 . They did not react with other taxonomically related Gram-negative organisms tested . The predominant isotype was immunoglobulin (Ig) M, although IgG2a, IgG2b, and IgG3 were also obtained . The epitopes identified by these mAbs were resistant to proteinase K treatment and boiling in the presence of sodium dodecyl sulfate and reducing conditions; however, they were sensitive to sodium periodate treatment . Enhanced chemiluminescence-immunodetection assay showed that mAbs could be divided in two groups according to the patterns of immunoreaction observed . Group 1 (mAbs 3E10, 4B7, 9H5 and 11C3) recognized a ladder-like banding profile consistent with the O antigen of lipopolysaccharide (LPS) from smooth strains . Group II (mAbs 3B10 and 9H1) recognized a long smear of high molecular weight which ranged from 60 to 200 kDa . The mAbs were tested against 96 field isolates belonging to serotypes 1, 5, 9, 11 and 12, which had previously been classified by a combination of serological techniques based on polyclonal rabbit sera (counterimmunoelectrophoresis, immunodiffusion and coagglutination) . The panel of mAbs identified all isolates of serotypes 9 and 11, but only 66% of those belonging to serotype 1 . This may suggest the existence of antigenic heterogeneity among isolates of A . pleuropneumoniae serotype 1 . These mAbs reacted with epitopes common to serotypes 1, 9 and 11 of Actinobacillus pleuropneumoniae which were located on the O antigen of LPS.

Proc Natl Acad Sci U S A, 1996 Dec 24, 93(26), 15086 - 91
Identification of a novel selD homolog from eukaryotes, bacteria, and archaea: is there an autoregulatory mechanism in selenocysteine metabolism?
Guimaraes MJ, Peterson D, Vicari A, Cocks BG, Copeland NG, Gilbert DJ, Jenkins NA, Ferrick DA, Kastelein RA, Bazan JF, Zlotnik A.
Escherichia coli selenophosphate synthetase (SPS, the selD gene product) catalyzes the production of monoselenophosphate, the selenium donor compound required for synthesis of selenocysteine (Sec) and seleno-tRNAs . We report the molecular cloning of human and mouse homologs of the selD gene, designated Sps2, which contains an in-frame TGA codon at a site corresponding to the enzyme's putative active site . These sequences allow the identification of selD gene homologs in the genomes of the bacterium Haemophilus influenzae and the archaeon Methanococcus jannaschii, which had been previously misinterpreted due to their in-frame TGA codon . Sps2 mRNA levels are elevated in organs previously implicated in the synthesis of selenoproteins and in active sites of blood cell development . In addition, we show that Sps2 mRNA is up-regulated upon activation of T lymphocytes and have mapped the Sps2 gene to mouse chromosome 7 . Using the mouse gene isolated from the hematopoietic cell line FDCPmixA4, we devised a construct for protein expression that results in the insertion of a FLAG tag sequence at the N terminus of the SPS2 protein . This strategy allowed us to document the readthrough of the in-frame TGA codon and the incorporation of 75Se into SPS2 . These results suggest the existence of an autoregulatory mechanism involving the incorporation of Sec into SPS2 that might be relevant to blood cell biology . This mechanism is likely to have been present in ancient life forms and conserved in a variety of living organisms from all domains of life.

J Theor Biol, 1996 Dec 21, 183(4), 455 - 69
Analysis of genomic patchiness of Haemophilus influenzae and Saccharomyces cerevisiae chromosomes; Lio P et al.; We have analysed some aspects of the primary structure of the chromosome of the prokaryote Haemophilus influenzae and of the eukaryote Saccharomyces cerevisiae that share the same G + C content . In particular, we have investigated genomic patchiness over the gene size level (10 Kb) and that patchiness due to long homogenous tracts . Long polypurine and polypyrmidine tracts that are largely over-represented in S . cerevisiae chromosomes and under-represented in H . influenzae, are responsible for a large fraction of long correlation signals . Generating mechanisms of long homogenous tracts are DNA replication slippage and duplication events that appear to be linked processes driving chromosome primary structure evolution.

Lancet, 1996 Dec 21-28, 348(9043), 1688 - 92
Randomised trial of the effect of co-administration with acellular pertussis DTP vaccine on immunogenicity of Haemophilus influenzae type b conjugate vaccine; Eskola J et al.; BACKGROUND: Inclusion of new vaccines in vaccination programmes for children would be easier if they could be combined with existing vaccines . Vaccines containing acellular pertussis in the diphtheria/tetanus/pertussis (DTP-a) combination are expected to replace the conventional whole-cell vaccines (DTP-w) . We tested the immunogenicity and safety of a combination of DTP-a with the Haemophilus influenzae type b (Hib) conjugate of Hib capsular polysaccharide and tetanus toxid (PRP-T), and inactivated poliovirus vaccine (i.p.v.) . METHODS: 120 infants were enrolled and randomised to four groups to receive DTP-a at ages 2, 4, and 6 months . At 4 and 6 months they also received Hib conjugate and i.p.v., either as separate injections or mixed with DTP-a . All injections were given intramuscularly in the anterolateral area of the thigh . Any reactions after each vaccination were noted by the parents . EIA was used to measure titres of diphtheria, tetanus, and pertussis antibodies, RIA for Hib anticapsular antibodies, and microneutralisation assay for poliovirus antibodies from serum samples collected at the ages of 2, 4, 6, and 7 months . FINDINGS: There were 30 infants in each group . Only mild adverse events were reported . There was a tendency towards slightly lower concentrations of filamentous haemagglutinin, tetanus, and poliovirus 1 antibodies when the vaccines were mixed . However, there was a more pronounced difference (p = 4 x 10(-6)) in Hib antibodies between groups receiving Hib capsular polysaccharide mixed with DTP-a (geometric mean concentrations 0.37 microgram/mL and 0.56 microgram/mL) compared with groups receiving the vaccines separately (3.10 micrograms/mL and 3.94 micrograms/mL) . INTERPRETATION: Administration of premixed DTP-a, Hib conjugate, and i.p.v . affect the immune response significantly . The mechanism of this interference is not clear . The immunogenicity of all antigens must be tested before new combinations can be accepted for vaccination programmes for infants.

FEMS Microbiol Lett, 1996 Dec 15, 145(3), 325 - 31
The identification a novel gene required for lipopolysaccharide biosynthesis by Haemophilus influenzae RM7004, using transposon Tn916 mutagenesis; High NJ et al.; Mutagenesis with the transposon Tn916 was used as a strategy to identify genes required for synthesis of the Gal alpha (1-4) beta Gal component of Haemophilus influenzae strain RM7004 lipopolysaccharide . Insertion of Tn916 into an open reading frame (ORF) encoding a protein with 75% homology to the Escherichia coli methionine related protein (Mrp) is described . Mutations in mrp resulted in loss of reactivity with monoclonal antibody (mAb) 4C4, which recognises Gal alpha (1-4) beta Gal, and expression of LPS with a different electrophoretic profile to that of wild-type RM7004 . An unexpected feature of this mutation was that it appeared to influence the number of copies of 5'-CAAT-3' present in lic2A, a gene which is also required for biosynthesis and phase variable expression of the Gal alpha (1-4) beta Gal LPS epitope.

Gene, 1996 Dec 12, 183(1-2), 35 - 40
Cloning and sequencing of the gene encoding the Cu,Zn-superoxide dismutase of Haemophilus ducreyi; Stevens MK et al.; The sodC gene of Haemophilus ducreyi was cloned and sequenced . The deduced amino acid sequence of this protein exhibited 71.6% identity and 81.8% similarity to the H . influenzae and H . parainfluenzae copper (Cu), zinc (Zn)-superoxide dismutase (SOD) enzymes . This gene was localized to a 2.2-kb H . ducreyi chromosomal DNA insert in plasmid pHdSOD . SOD activity was expressed in cell-free extracts of Escherichia coli containing the recombinant plasmid pHdSOD and was localized to the periplasmic space . The Cu,Zn-SOD produced by the H . ducreyi sodC gene did not complement the aerobic growth defect of an E . coli SOD-deficient mutant.

Vet Rec, 1996 Dec 7, 139(23), 561 - 3
Otitis in a weaned pig: a new pathological role for Actinobacillus (Haemophilus) pleuropneumoniae; Duff JP et al.; A weaned pig exhibiting clinical signs characteristic of a vestibular syndrome (abnormal head tilt and abnormal gait including a tendency to circle) was examined as a representative example of a condition which, in a 12-month period, affected approximately 400 other pigs on the farm of origin . The condition caused significant morbidity and financial loss but the directly attributable mortality was negligible . Post mortem examination revealed a severe suppurative, left-sided otitis media and interna . Mixed growths of bacteria, including Actinobacillus pleuropneumoniae, were isolated from the pus from the middle ear . Otitis caused by mixed bacterial infections has been described previously in pigs and cattle but infection of the porcine middle ear cavity by A pleuropneumoniae is considered to be a new pathological role for this organism . Other findings in this case indicated that otitis media had followed from bacterial colonisation of the auditory (Eustachian) tube . The condition was effectively controlled by prophylactic antibiotic therapy . The authors consider that vestibular syndrome due to otitis media and interna, whether caused by infection with A pleuropneumoniae or not, is probably a relatively common condition in England but its significance may be underestimated and for this reason it may be underrecorded.

Int J Pediatr Otorhinolaryngol, 1996 Dec 5, 38(1), 41 - 51
Airway endoscopy in the interdisciplinary management of acute epiglottitis; Damm M et al.; INTRODUCTION: Acute epiglottitis (AE) continues to cause life-threatening airway obstruction in children . The aim of this retrospective study was to evaluate deficiencies in the management of AE, to clarify the role of airway endoscopy, and to establish the cause of death in this disease . MATERIAL: The subjects of our retrospective study were 24 children with AE treated in the paediatric intensive care unit (PICU) of the University of Cologne between 1980 and 1994 . The records of all patients were reviewed . RESULTS: Laryngoscopy with fibreoptic or small rigid endoscopes allowed accurate diagnosis of AE, particularly in patients presented with atypical clinical signs . Furthermore, endoscopic information of the degree of inflammation were helpful in the critical decision, whether artificial airway was required . The number of patients managed without intubation increased (8% vs . 45%) since airway endoscopy became available in 1989 . No evidence of beta-lactamase-producing strains of Haemophilus influenzae was observed . The most remarkable finding was the high mortality in AE (12.5%) . Affected children were admitted in poor post-hypoxia conditions following outdoor cardiorespiratory arrest . CONCLUSION: The most decisive factor to decrease mortality seems to be timely appropriate presentation at referring centres, if AE is suspected or stridor remains unclear . Fibreoptic airway endoscopy represents a major step forward in the management of acute epiglottitis, and this requires close interdisciplinary collaboration between paediatricians, anaesthetists and otorhinolaryngologists.

Antimicrob Agents Chemother, 1996 Dec, 40(12), 2884 - 6
Prevalence of antimicrobial resistance among 723 outpatient clinical isolates of Moraxella catarrhalis in the United States in 1994 and 1995: results of a 30-center national surveillance study; Doern GV et al.; Seven hundred twenty-three isolates of Moraxella catarrhalis obtained from outpatients with a variety of infections in 30 medical centers in the United States between 1 November 1994 and 30 April 1995 were characterized in a central laboratory . The overall rate of beta-lactamase production was 95.3% . When the National Committee for Clinical Laboratory Standards MIC interpretive breakpoints for Haemophilus influenzae were applied, percentages of strains found to be susceptible to selected oral antimicrobial agents were as follows: azithromycin, clarithromycin, and erythromycin, 100%; tetracycline and chloramphenicol, 100%; amoxicillin-clavulanate, 100%; cefixime, 99.3%; cefpodoxime, 99.0%; cefaclor, 99.4%; loracarbef, 99.0%; cefuroxime, 98.5%; cefprozil, 94.3%; and trimethoprim-sulfamethoxazole, 93.5%.

Antimicrob Agents Chemother, 1996 Dec, 40(12), 2796 - 801
Postantibiotic and post-beta-lactamase inhibitor effects of amoxicillin plus clavulanate; Thorburn CE et al.; The postantibiotic effect (PAE) of amoxicillin-clavulanate was studied for strains of Staphylococcus aureus, Haemophilus influenzae, Moraxella catarrhalis, Klebsiella pneumoniae, and Escherichia coli . A PAE of approximately 2 h was seen for beta-lactamase-positive and -negative strains of S . aureus following 2 h of exposure to twice the MIC and did not increase at 16 times the MIC . The PAE observed with H . influenzae was clearly related to the growth rate of the organism . A PAE of 0.8 h was found for amoxicillin (four times the MIC) against a beta-lactamase-negative strain of H . influenzae (generation time, 26.3 min) and a PAE of 1.74 h was found for amoxicillin-clavulanate (twice the MIC) against a beta-lactamase-positive strain (generation time, 32.2 min) . When the beta-lactamase-positive strain was growing more slowly (generation time, 120 min), the PAE of amoxicillin-clavulanate increased to > 3.32 h . The PAE of amoxicillin-clavulanate at 2/1 micrograms/ml on a beta-lactamase-producing strain of M . catarrhalis was > 2.9 h, and, as expected, the PAEs of twice the MIC on K . pneumoniae and E . coli were generally short (< 1 h) . The post-beta-lactamase inhibitor effect (PLIE), determined after removal of only clavulanate, was also examined for beta-lactamase-positive strains . This was more prolonged (approximately 3 to 4 h) than the corresponding PAE for S . aureus, H . influenzae, and M . catarrhalis . The PLIE was related to the amount of beta-lactamase produced and required the presence of amoxicillin in the initial exposure period . These data may have implications for reducing the dosage of amoxicillin-clavulanate.

Hiroshima J Med Sci, 1996 Dec, 45(4), 113 - 7
Involvement of bacterial antigens in immunoglobulin A nephropathy; Hirabayashi A et al.; To investigate the involvement of bacterial antigens in Immunoglobulin A (IgA) nephropathy, we measured IgA, IgG and IgM antibodies to gram-negative Escherichia coli (E.coli) and Haemophilus influenzae (H.influenzae) by ELISA in 24 patients (11 males and 13 females) with IgA nephropathy and 22 normal controls (11 males and 11 females) . The titers of IgA and IgM antibodies for E.coli and H.influenzae were significantly higher in the IgA nephropathy group than in the controls . In addition, IgA and IgM antibody titers for E.coli and H.influenzae showed a significant positive correlation with serum IgA and IgM levels . These findings suggest that subclinical infection by these bacteria stimulates IgA production and that this may be a factor in the development and progression of IgA nephropathy.

Minerva Pediatr, 1996 Dec, 48(12), 551 - 69
{Current problems in the treatment of bacterial meningitis}; Principi N; This paper on bacterial meningitis looks at aspects inherent in the aetiology and mechanisms underlying neurological damage and pharmacological treatment . Streptococcus pneumoniae, Haemophilus influenzae type b and Neisseria meningitidis are the pathogens most commonly responsible and are able to colonise the host's respiratory mucosae, invade the vascular space, cross the haematoliquoral barrier and survive in the cerebrospinal fluid . The presence of germs in the subarachnoid spaces leads to the onset of inflammation and neurological damage . The most often used pharmacological treatments include, apart from antibiotics, anti-inflammatory drugs (although we have clinical data for corticosteroids only), pentoxyphillin and monoclonal antibodies . Initially empiric, antibiotic therapy is based on the use of drugs that act against the probable pathogenic agents, are capable of surmounting the haematoliquoral barrier and are well tolerated . Prior to the Eighties, the antibiotic of choice was ampicillin associated or otherwise with aminoglycosides . Subsequently, the availability of new drugs (cefotaxime and ceftriaxone) and the appearance of resistance led to changes in therapeutic protocols . Of the carbapenemics, wide spectrum antibiotics with high resistance to beta lactamase, imipenem /cilastatin proved effective although there was a high risk of inducing convulsions in patients with previous neurological damage or kidney failure . Meropenem was able to surmount the haematoliquoral barrier in sufficient concentrations and was well tolerated in patients with prior neurological changes . It has proved effective in clinical studies carried out up to the present.

Int J Epidemiol, 1996 Dec, 25(6), 1280 - 5
Risk factors for invasive Haemophilus influenzae disease among children 2-16 years of age in the vaccine era, Switzerland 1991-1993 . The Swiss H . Influenzae Study Group; Muhlemann K et al.; BACKGROUND: Continued surveillance, and detailed investigation of direct and indirect effects of conjugated vaccines and risk factors for invasive H.influenzae serotype B (Hib) disease in the vaccine era are important . METHODS: 143 cases with invasive disease between 1991 and 1993 aged 2-16 years were selected retrospectively from a large incidence trend study . Controls (n = 336) were recruited from local vital registries and matched to cases for age, gender, and residence . Hib vaccination histories among study subjects and their siblings and other sociodemographic variables were obtained by questionnaires completed by the parents of these children . Adjusted odds ratio (OR) estimates were calculated by conditional logistic regression analysis . RESULTS: Most vaccinated subjects had received the Polysaccharide-Diphtheria Toxoid vaccine and estimated vaccine efficacy was high (95%; 95% confidence interval {CI} 60-99%) . Also, the results suggested that protection afforded by vaccination against Hib extended to the family members of vaccinated children . School attendance was found to be protective against invasive Hib disease (OR:0.33; CI:1.2-14.4) . Cases more often than controls reported suffering from asthma and allergies (OR:4.8; CI:1.2-19.4) . CONCLUSIONS: Post-licensure vaccine efficacy is high among children > or = 2 years of age . The observed association between asthma and epiglottitis is novel and deserves further investigation.

Comput Appl Biosci, 1996 Dec, 12(6), 519 - 24
Correspondence discriminant analysis: a multivariate method for comparing classes of protein and nucleic acid sequences; Perriere G et al.; This report describes two applications of a multivariate method for studying classes of nucleotide or protein sequences: correspondence discriminant analysis (CDA) . The first example is the discrimination between Escherichia coli proteins according to their subcellular location (membrane, cytoplasm and periplasm) . The high resolution of the method made it possible to predict the subcellular location of E.coli proteins for whom this information is not known . The second example is discrimination between the coding sequences of leading and lagging strands in four bacteria: Mycoplasma genitalium, Haemophilus influenzae, E.coli and Bacillus subtilis . The programs used for computing the analysis are integrated in a publicly available package that runs on MacOS 7.x or Windows 95 operating systems biomserv.univ-lyonl.fr/ADE-4.html) . These programs are also accessible through our World Wide Web server biomserv.univ-lyonl.fr/Net Mul.html).

J Trop Pediatr, 1996 Dec, 42(6), 318 - 23
Variation in the pattern of bacterial infection in patients with sickle cell disease requiring admission; Akuse RM; In order to determine the prevalence and pattern of bacterial infections in children with sickle cell disease (SCD) admitted with acute illness, a prospective study of 304 sicklers aged 3 months to 15 years was carried out over 1 year in the emergency pediatric unit of ABU Teaching Hospital Kaduma . Initial blood and urine cultures were obtained in all cases and other cultures were performed as determined by the patients' clinical condition . Almost 60 per cent of the patients had positive bacterial cultures with gram negative organisms accounting for 55 per cent of them, but the single most predominant organism isolated was Staphylococcus aureus . Of note was the strikingly low isolation rate of Streptococcus pneumoniae, Haemophilus influenzae and Neisseria meningitides . Possible reasons for this difference from the majority of reports from the western World are advanced and the implications discussed, especially as regards vaccination programmes in sickle cell disease and initial antibiotic treatment of those with acute illness.

Microbiology, 1996 Dec, 142 ( Pt 12), 3505 - 13
Conservation and antigenic cross-reactivity of the transferrin-binding proteins of Haemophilus influenzae, Actinobacillus pleuropneumoniae and Neisseria meningitidis; Holland J et al.; Haemophilus influenzae acquires iron from the iron-transporting glycoprotein transferrin via a receptor-mediated process . This involves two outer-membrane transferrin-binding proteins (Tbps) termed Tbp1 and Tbp2 which show considerable preference for the human form of transferrin . Since the Tbps are attracting considerable attention as potential vaccine components, we used transferrin affinity chromatography to examine their conservation amongst 28 H . influenzae type b strains belonging to different outer-membrane-protein subtypes as well as six non-typable strains . Whole cells of all type b and non-typable strains examined bound human transferrin; whilst most strains possessed a Tbp1 of approximately 105 kDa, the molecular mass of Tbp2 varied from 79 to 94 kDa . Antisera raised against affinity-purified native H . influenzae Tbp1/Tbp2 receptor complex cross-reacted on Western blots with the respective Tbps of all the Haemophilus strains examined . When used to probe Neisseria meningitidis Tbps, sera from each of four mice immunized with the Haemophilus Tbp1/2 complex recognized the 68 kDa Tbp2 of N . meningitidis strain B16B6 but not the 78 kDa Tbp2 of N . meningitidis strain 70942 . Serum from one mouse also reacted weakly with Tbp1 of strain B16B6 . Apart from a weak reaction with the Tbp2 of a serotype 5 strain, this mouse antiserum failed to recognize the Tbps of the porcine pathogen A . pleuropneumoniae . However, a monospecific polyclonal antiserum raised against the denatured Tbp2 of Neisseria meningitidis B16B6 recognized the Tbps of all Haemophilus and Actinobacillus strains examined . Since H . influenzae forms part of the natural flora of the upper respiratory tract, human sera were screened for the presence of antibodies to the Tbps . Sera from healthy adults contained antibodies which recognized both Tbp1 and Tbp2 from H . influenzae but not N . meningitidis . Convalescent sera from meningococcal meningitis patients contained antibodies which, on Western blots, recognized the Tbps2s of both pathogens . These data demonstrate the existence of shared epitopes on the Tbps of H . influenzae, N . meningitidis and A . pleuropneumoniae despite their transferrin species specificity.

Curr Opin Genet Dev, 1996 Dec, 6(6), 757 - 62
Complete genome sequences of cellular life forms: glimpses of theoretical evolutionary genomics; Koonin EV et al.; The availability of complete genome sequences of cellular life forms creates the opportunity to explore the functional content of the genomes and evolutionary relationships between them at a new qualitative level . With the advent of these sequences, the construction of a minimal gene set sufficient for sustaining cellular life and reconstruction of the genome of the last common ancestor of bacteria, eukaryotes, and archaea become realistic, albeit challenging, research projects . A version of the minimal gene set for modern-type cellular life derived by comparative analysis of two bacterial genomes, those of Haemophilus influenzae and Mycoplasma genitalium, consists of approximately 250 genes . A comparison of the protein sequences encoded in these genes with those of the proteins encoded in the complete yeast genome suggests that the last common ancestor of all extant life might have had an RNA genome.

Mol Microbiol, 1996 Dec, 22(5), 951 - 65
Use of the complete genome sequence information of Haemophilus influenzae strain Rd to investigate lipopolysaccharide biosynthesis; Hood DW et al.; The availability of the complete 1.83-megabase-pair sequence of the Haemophilus influenzae strain Rd genome has facilitated significant progress in investigating the biology of H.influenzae lipopolysaccharide (LPS), a major virulence determinant of this human pathogen . By searching the H . influenzae genomic database, with sequences of known LPS biosynthetic genes from other organisms, we identified and then cloned 25 candidate LPS genes . Construction of mutant strains and characterization of the LPS by reactivity with monoclonal antibodies, PAGE fractionation patterns and electrospray mass spectrometry comparative analysis have confirmed a potential role in LPS biosynthesis for the majority of these candidate genes . Virulence studies in the infant rat have allowed us to estimate the minimal LPS structure required for intravascular dissemination . This study is one of the first to demonstrate the rapidity, economy and completeness with which novel biological information can be accessed once the complete genome sequence of an organism is available.

Microb Pathog, 1996 Dec, 21(6), 463 - 70
The effect of mutations in genes required for lipopolysaccharide synthesis on Haemophilus influenzae type b colonization of human nasopharyngeal tissue; Jackson AD et al.; Haemophilus influenzae type b (Hib) is an upper respiratory tract commensal that can cause invasive disease, particularly in young children . Lipopolysaccharide (LPS) has been implicated as a major virulence determinant of Hib, and changes in LPS structure may influence bacterial interactions with the respiratory mucosa . We have examined the effect of variations in LPS on the interaction of Hib with human nasal turbinate tissue maintained in an organ culture model with an air-interface, by using isogenic derivatives of strains RM153 (Eagan) and RM7004 expressing truncated LPS due to mutations in genes contained within the chromosomal loci lic1 and lic2 (lic1lic2) or in the galE and galK genes (galEK) . Tissue was infected with an inoculating dose of 2.3-3.3 x 10(6) colony forming units (cfu) in 2 microliters of PBS and maintained for 24 h . By scanning electron microscopy the percentage of the organ culture surface exhibiting epithelial damage increased from 5.3 +/- 1.4 in controls to 12.5 +/- 6.4-26.3 +/- 9.1 following infection, with no significant difference between parent strains and their derivatives . There was significant bacterial tropism for mucus, and to a lesser extent damaged cells, which was not influenced by the LPS phenotype . All strains caused separation of epithelial cells, adhered to non-luminal cell surfaces, and invaded the epithelium intercellularly . We conclude that Hib associated with mucus and damaged epithelium, and infrequently with normal epithelium, but changes in the LPS phenotype did not affect the interaction between Hib and the mucosal surface of human nasal turbinate tissue.

Am J Respir Crit Care Med, 1996 Dec, 154(6 Pt 1), 1788 - 93
Inhibition by dextran of Pseudomonas aeruginosa adherence to epithelial cells; Barghouthi S et al.; Adherence of Pseudomonas aeruginosa to cells of the respiratory tract of patients with cystic fibrosis (CF) appears to be a necessary precondition for colonization and infection . To date no effective anti-adhesive strategy has been devised for preventing P . aeruginosa infection in these vulnerable hosts . The purpose of these studies was to evaluate the potential for preventing adhesion of P . aeruginosa to epithelial cells with dextran . Dextran (3,000-70,000 MW) inhibited adhesion of P . aeruginosa to buccal and A549 pulmonary epithelial cells; the 3,000 MW compound, at 10 mM was most inhibitory . Adhesion was inhibited optimally at pH 7.4 and was independent of charge; dextran and dextran sulfate were equally inhibitory . Dextran was most inhibitory if added to the epithelial cells before the P . aeruginosa; adhesion was reversed only minimally by adding dextran after the bacteria were bound . The inhibitory effect appeared to be nonspecific because other neutral polysaccharides (glycogen and mannan) were also inhibitory, dextran blocked attachment of other respiratory tract pathogens (Staphylococcus aureus, Group A streptococcus, and Haemophilus influenzae), and because dextran did not bind specifically to bacteria or to epithelial cells . Dextran is an inexpensive and nontoxic agent and may be useful in patients with CF to prevent colonization and infection with P . aeruginosa.

Pediatr Infect Dis J, 1996 Dec, 15(12), 1097 - 101
Persistence of antibody responses to Haemophilus influenzae type b polysaccharide conjugate vaccine in children with vertically acquired human immunodeficiency virus infection; Gibb D et al.; BACKGROUND: Recurrent bacterial sepsis is common in pediatric HIV infection and immunization against Haemophilus influenzae type b (Hib) is recommended . Long term persistence of anti-Hib antibody and the need for, or timing of, a booster dose has not been adequately studied . METHODS: Immunogenicity during a 12-month period following immunization with Hib-tetanus conjugate vaccine (ACT-HIB; Merieux) was evaluated in 48 vertically HIV-infected children and 36 uninfected children, born to HIV-positive mothers . A titer of anti-Hib polysaccharide antibody of > or = 0.15 microgram/ml was considered to indicate short term and > or = 1 microgram/ml long term protection . RESULTS: At 1 month postvaccination 36 (100%) uninfected and 42 (88%) HIV-infected children achieved titers of > or = 1 microgram/ml . However, by 1 year titers had dropped below this value in 18 (43%) infected compared with only 4 (11%) uninfected children (chi square, 9.7; P = 0.002) . Although the rate of fall of antibody titer was greater in uninfected than in infected children, this was no longer the case after adjustment for the 1-month postimmunization titer . The rate of antibody titer decline was not significantly related to HIV disease status or to either the age-related CD4 count at the time of immunization or the change in age-adjusted CD4 count during the 12 months after immunization . CONCLUSIONS: Not only was the initial antibody response to Hib conjugate vaccine decreased in children with HIV infection and AIDS but also 1 year later only 57% of the initial responders had persisting titers above the level associated with long term protection . The need for reimmunization of children with HIV infection against Hib requires further evaluation.

J Bacteriol, 1996 Dec, 178(23), 6960 - 7
Cloning and genetic characterization of Helicobacter pylori catalase and construction of a catalase-deficient mutant strain; Odenbreit S et al.; The N-terminal sequence of a protein, originally described as an adhesin of Helicobacter pylori, was used in an oligonucleotide-based screening procedure of an H . pylori plasmid library in Escherichia coli . Five independent plasmid clones were isolated, all mapping to the same chromosomal region and encoding the H . pylori catalase . The gene, designated katA, comprises 1,518 nucleotides and encodes a putative protein of 505 amino acids with a predicted Mr of 58,599 . A second open reading frame, orf2, encoding a putative 32,715-Da protein of unknown function, follows katA . The transcriptional start site of katA mRNA was determined, but no typical consensus promoter sequence was present . A potential binding site for the Fur protein is located upstream of katA . When introduced into the catalase-deficient E . coli double-mutant UM255, the cloned gene readily complemented E . coli for catalase activity . H . pylori KatA is highly homologous to catalases in both prokaryotes and eukaryotes, with the highest homology being shown to Bordetella pertussis (64.9%), Bacteroides fragilis (59.8%), and Haemophilus influenzae (57.9%) catalases . Transposon insertion mutants were generated in three independent H . pylori strains by TnMax5-mediated transposon shuttle mutagenesis . In contrast to the wild-type strains, no significant catalase-specific enzymatic activity could be detected in the mutant strains, consistent with the fact that no additional katA-homologous gene copies were found in the H . pylori chromosome . No significant difference between wild-type and mutant strains for binding to epithelial cells was apparent, suggesting that KatA is not involved in H . pylori adhesion . The cloning and genetic characterization of katA are essential steps for further investigation of the role of catalase in the defense of H . pylori against oxygen-dependent killing mechanisms by polymorphonuclear granulocytes, a process not well understood for this chronically persisting pathogen.

Ann Emerg Med, 1996 Dec, 28(6), 617 - 20
Changing bacteriology of periorbital cellulitis; Schwartz GR et al.; STUDY OBJECTIVE: Bacteremic periorbital cellulitis has traditionally been associated with Haemophilus influenzae infection, and the recommended diagnostic evaluation in young children includes blood culture and cerebrospinal fluid (CSF) analysis . The objectives of this study were to examine in pediatric patients with periorbital cellulitis (1) the prevalence of H influenzae bacteremia in the era of vaccination for H influenzae type B (HIB) and (2) the yield of routine CSF analysis . METHODS: This was a retrospective case series of children aged 2 months to 17 years with a final discharge diagnosis of periorbital cellulitis who were treated from 1986 through 1994 at an urban university referral hospital . The prevalence of bacteremia and meningitis was obtained from chart review . RESULTS: Forty-nine children were enrolled, of whom 3 were treated as outpatients . The mean age was 36 months (median, 19 months) . A blood culture was obtained for 92% of the patients . Five patients (10%; 95% confidence interval, 3% to 22%) had a positive blood culture (four streptococcal species, one H influenzae) . The patient with H influenzae bacteremia was treated in 1987 and had not had the HIB vaccine . CSF was analyzed for 41% of the patients, and none had an abnormal cell count or a positive culture . Sinusitis was diagnosed radiographically in 19% of the subjects . CONCLUSION: Streptococcal organisms are the most common cause of bacteremia associated with periorbital cellulitis in the post-HIB vaccination era . H influenzae bacteremia is now a rare occurrence . Meningitis is uncommon, and lumbar puncture may not be mandatory in well-appearing children . Sinusitis is common and was probably underdiagnosed in our series because most children were not evaluated radiographically . Outpatient management may be indicated in selected children.

Clin Infect Dis, 1996 Dec, 23(6), 1305 - 7
Pott's puffy tumor due to Haemophilus influenzae: case report and review; Verbon A et al.; Osteomyelitis of the frontal bone may be associated with a purulent collection under the periosteum, causing swelling and edema over the forehead, a condition known as Pott's puffy tumor . We describe an 83-year-old man with a Pott's puffy tumor due to Haemophilus influenzae that was successfully treated with surgery and antibiotics . A review of 22 cases of Pott's puffy tumor shows that this condition usually occurs in children, is spread from frontal or ethmoid sinusitis, and is usually due to streptococci, staphylococci, or anaerobes . Suppurative complications such as epidural, subdural, and intracerebral abscesses are common . Only seven cases of Pott's puffy tumor in adults have been reported, and only one of these cases was caused by H . influenzae . Surgical drainage and antibiotic therapy remain standard therapy for this condition.

Clin Infect Dis, 1996 Dec, 23(6), 1240 - 5
Serological study of responses to selected pathogens causing respiratory tract infection in the institutionalized elderly; Orr PH et al.; In a prospective 2-year study, serological responses to selected pathogens were analyzed in 224 episodes of fever attributable to respiratory tract infection (51.8%) or of unknown source (48.2%) in 131 residents of two long-term-care facilities . A serological response was identified in 45 episodes (20.1%): Chlamydia pneumoniae (14 episodes), Haemophilus influenzae type b (1), influenza virus type A (14), respiratory syncytial virus (RSV;2), parainfluenza virus type 3 (7), C . pneumoniae and H . influenzae (3), C . pneumoniae and influenza virus type A (2), C . pneumoniae and RSV (1), and C . pneumoniae and parainfluenza virus type 3 (1) . No serological responses to Chlamydia psittaci, Chlamydia trachomatis, parainfluenza virus types 1 and 2, influenza virus type B, or Mycoplasma pneumoniae were seen . Vaccination did not affect the duration of fever in those residents with serologically confirmed influenza A . Serologically confirmed C . pneumoniae infection was detected in 9.4% of all febrile episodes . Serological responses to a second agent were detected in 33% of the patients with C . pneumoniae infections, and these dual infections were associated with an underlying malignancy (P = .02) . C . pneumoniae should be recognized as a potential pathogen when choosing empirical antimicrobial therapy for respiratory tract infection in residents of long-term-care facilities.

Infect Immun, 1996 Dec, 64(12), 5187 - 92
Immunization with outer membrane protein P6 from nontypeable Haemophilus influenzae induces bactericidal antibody and affords protection in the chinchilla model of otitis media; DeMaria TF et al.; The role of nontypeable Haemophilus influenzae (NTHi) outer membrane protein (OMP) P6 in the pathogenesis of otitis media (OM) has not been defined . OMPs, fimbriae, pili, and lipooligosaccharide are several types of surface antigens of NTHi that are currently being evaluated as potential vaccine candidates . P6 is antigenically conserved among both nontypeable and type b H . influenzae strains and elicits bactericidal as well as protective antibodies; however, initial evaluation of a vaccine mixture of P6 combined with other NTHi OMPs failed to induce bactericidal antibody or protection in the chinchilla model of OM . We undertook an assessment of the ability of immunization with isolated P6 lipoprotein alone to confer protection . Chinchillas were immunized with P6 and challenged 10 days after the final immunization with either 3 x 10(3) CFU of NTHi delivered directly into the middle ear to induce OM or 5 x 10(8) CFU of NTHi delivered intranasally to establish nasopharyngeal colonization . All immunized animals responded with elevated serum titers of anti-P6 antibody, which also demonstrated bactericidal activity against homologous as well as a heterologous NTHi isolate . By 14 days post-transbullar challenge, the number of chinchillas with middle ear fluid and the incidence of NTHi culture-positive middle ear fluids were reduced 48 and 51%, respectively, in the P6-immunized chinchillas relative to the sham-immunized cohort . Nasopharyngeal colonization levels were comparable in the two cohorts . These data demonstrate that active immunization with P6 results in the production of NTHi-specific bactericidal antibody in the chinchilla and also affords a reduction in the incidence of NTHi-induced OM; however, parenteral immunization does not appear to affect the extent or duration of nasopharyngeal colonization by NTHi.

J Infect Dis, 1996 Dec, 174(6), 1345 - 7
Long-term persistence of ciprofloxacin-resistant Haemophilus influenzae in patients with cystic fibrosis; Campos J et al.; Ciprofloxacin has been a major advance in the treatment of chronic respiratory infections . Three patients with cystic fibrosis and colonized by 5 nontypeable Haemophilus influenzae strains exhibiting low- (MIC, 2 microg/mL) and high-level ciprofloxacin resistance (MICs, 16-32 microg/mL) are described . The patients had received several courses of ciprofloxacin . These MICs represent a decrease in ciprofloxacin susceptibility of 200-3200 times . Molecular epidemiologic methods demonstrated that 2 patients were chronically colonized by their own ciprofloxacin-resistant strains for > or = 15-17 months . Three strains showed simultaneous resistance to ampicillin and chloramphenicol by enzyme inactivation, and 2 had ampicillin resistance without beta-lactamase activity . These data suggest that the emergence and long-term persistence of ciprofloxacin-resistant H . influenzae in patients with cystic fibrosis can be a consequence of antibiotic treatment.

J Infect Dis, 1996 Dec, 174(6), 1341 - 4
No adverse impact on protection against pertussis from combined administration of Haemophilus influenza type b conjugate and diphtheria-tetanus toxoid-pertussis vaccines in the same syringe; Levine OS et al.; To assess whether combining a Haemophilus influenzae type b conjugate vaccine (PRP-T) and diphtheria-tetanus toxoid-pertussis (DTP) vaccine in a single syringe would impact adversely the antibody response and clinical protection conferred by pertussis vaccine, surveillance and a nested serosurvey were conducted among infants in a large-scale evaluation of PRP-T in Santiago . Infants received either combined PRP-T/DTP or DTP only at 2, 4, and 6 months of age . At 8 months, pertussis agglutinin, anti-pertussis toxin, and anti-filamentous hemagglutinin antibody levels in the PRP-T/DTP (137.7, 23.1, and 12.2, respectively) and DTP (142.9, 20.6, and 13.0, respectively) groups were comparable . The incidence of pertussis was similar among infants assigned to health centers administering combined PRP-T/DTP and those administering DTP alone (13.1 vs . 12.2 cases/10(5) child-years) . Combined PRP-T/DTP vaccine did not diminish protection against pertussis.

J Infect Dis, 1996 Dec, 174(6), 1337 - 40
Mechanism of antibody-mediated reduction of nasopharyngeal colonization by Haemophilus influenzae type b studied in an infant rat model; Kauppi-Korkeila M et al.; The mechanism of antibody-mediated reduction of Haemophilus influenzae type b (Hib) carriage was studied in the infant rat colonization model . Monoclonal Hib polysaccharide (PS) antibody (MAb) given intranasally or intraperitoneally and human secretory anti-Hib PS IgA given intranasally inhibited colonization by Hib during the entire follow-up period (2-48 h after challenge) but did not affect colonization by Hi, a noncapsulated variant of Hib . F(ab')2 fragments, prepared from the MAb or from human serum anti-Hib IgG reduced Hib colonization as efficiently as the uncleaved molecules . Complement depletion by cobra venom treatment had no effect on the antibody-mediated reduction of Hib colonization . These results indicate that Fc-mediated activities of immunoglobulins are not essential in the reduction of Hib colonization . Instead, antibodies to Hib most likely reduce colonization by a direct effect on growth of the bacteria or their adherence to the nasopharyngeal mucosa.

Gene, 1996 Nov 28, 181(1-2), 77 - 83
Sequence and transcriptional analysis of clpX, a class-III heat-shock gene of Bacillus subtilis; Gerth U et al.; The nucleotide sequence of clpX, which is localized between the tig (trigger factor) and the lon (ATP-dependent protease) genes at 245 degrees on the standard Bacillus subtilis (Bs) genetic map, was determined . The putative clpX gene codes for a 46-kDa protein of 421 amino acid (aa) residues . A comparison of the deduced aa sequence with those of the recently described bacterial clpX gene products from Synechocystis sp., Escherichia coli (Ec), Haemophilus influenzae and Azotobacter vinelandii revealed strong similarities . However, in contrast to Ec, clpX and clpP of Bs are located at different loci on the chromosome and are transcribed as monocistronic genes . A heat-inducible sigma A-like promoter was mapped upstream of the clpX structural gene, but no CIRCE element, characteristic of class-I heat-shock genes (e.g., groESL and dnaK), was found between the transcriptional and translational start sites . Although the majority of the heat-inducible general stress genes in Bs are under the control of the alternative sigma factor, sigma B, the heat induction of clpX appears to be sigma B-independent . The latter indicates that clpX belongs to class-III heat-inducible genes.

Gene, 1996 Nov 21, 180(1-2), 49 - 56
Identification of a gene, pilF, required for type 4 fimbrial biogenesis and twitching motility in Pseudomonas aeruginosa; Watson AA et al.; Many bacterial pathogens produce a class of surface structures called type 4 fimbriae . In Pseudomonas aeruginosa these fimbriae are responsible for adhesion and translocation across host epithelial surfaces . We have identified a novel gene involved in the complex process of type 4 fimbrial biogenesis . This gene, termed pilF, is located on SpeI fragment S at 30 min on the P . aeruginosa genomic map, which is the sixth region on the chromosome shown to contain a fimbrial-associated gene . The PilF protein has a predicted M(r) of 22402, and together with a highly homologous upstream ORF shares a chromosomal arrangement similar to that found in Haemophilus influenzae . A pilF mutant is blocked in the export/assembly of the fimbrial subunit PilA, and accumulates this protein in the membrane fraction . Complementation studies indicate that the cloned pilF gene is able to restore the expression of surface fimbriae, twitching motility and susceptibility to fimbrial-specific bacteriophage.

MMWR Morb Mortal Wkly Rep, 1996 Nov 15, 45(45), 993 - 5
FDA approval of a Haemophilus b Conjugate Vaccine combined by reconstitution with an acellular pertussis vaccine.
{Vaccination of premature infants}
Kristensen K, Lausen BF.

Paediatrisk klinik 2, Juliane Marie Centret, Rigshospitalet, KobenhavnThe article gives a review of the literature concerning immunization of preterm infants . The initial response to diphtheria-tetanus-whole-cell-pertussis vaccine and inactivated polio vaccine may be impaired in preterms, but after two and three doses the responses to these vaccines is comparable to those of term infants of the same chronological age . Likewise, the response to oral polio vaccine is normal and independent of gestational age after two and three doses in preterm infants . After two doses, the response to Haemophilus influenzae type b conjugate vaccine is impaired in preterm infants with very low gestational age, but comparable to that of term infants in preterms with higher gestational ages . After three doses the response to this vaccine is normal and unaffected by gestational age in preterms . Recombinant hepatitis b vaccine induces a reduced response in preterm infants even after three doses, and they may therefore need an extra dose of this vaccine . Studies of diphtheria-tetanus-whole-cell-pertussis vaccine and Haemophilus influenzae type b conjugate vaccine in preterm infants do not suggest that adverse effects occur more often or are more severe than those seen in term infants . The literature provides no background for postponement of immunizations in preterm infants.

Braz J Med Biol Res, 1996 Nov, 29(11), 1485 - 90
Monoclonal antibodies against conserved epitopes of a 46-kDa protein from Neisseria meningitidis; Schuch MC et al.; The purpose of the present study was to generate monoclonal antibodies (mAbs) against conserved epitopes of B meningococcus which could be applicable to the immunoscreening of bacterial meningitis . Three mAbs reactive to a 46-kDa protein conserved in eight sero-groups and several sero(sub)types of Neisseria meningitidis were selected for the present study . No reaction was detected with whole-cell lysates of Staphylococcus aureus . Streptococcus pneumoniae, Haemophilus influenzae type b or Escherichia coli . Two of these mAbs recognized 46-kDa epitopes in four other Neisseria spp, and the third, MC3.13, cross-reacted only with N . lactamica . All mAbs reacted with whole-cell lysates from a N . meningitidis mutant strain lacking the class 1 outer membrane protein (43-47 kDa) . Immunoelectron microscopy revealed a cytoplasmic location for the 46-kDa protein . The MC3.13 monoclonal antibody is potentially applicable to a rapid screening of bacterial meningitis.

Acta Otorrinolaringol Esp, 1996 Nov-Dec, 47(6), 491 - 4
{Acute epiglottitis in the adult}; Lahoz Zamarro MT et al.; Epiglottitis is a well-defined syndrome caused by Haemophilus influenzae type B in children . It also may affect adults and has an unpredictable clinical course, sometimes complicated with airway obstruction . Two cases of adult epiglottitis are reported . The first responded favorably to medical treatment and the second required emergency airway surgery . Indicators of poor prognosis are described . Epiglottitis should be considered in the differential diagnosis as a potentially lethal cause of sore throat.

An Esp Pediatr, 1996 Nov, 45(5), 487 - 92
{Serum antibodies against Haemophilus influenzae in children from the province of Valladolid (Spain)}; Blanco Quiros A et al.; OBJECTIVE: Invasive infections by Haemophilus influenzae type b (Hib) in children are increasing in Spain . Moreover, they are becoming more resistant to antibiotics . Currently, effective vaccines against Hib are available and their inclusion in the routine schedule of immunizations is discussed . Knowledge of the childhood serology appears to be useful in decision making . PATIENTS AND METHODS: Two hundred and thirty-five normal children, between 1 month and 15 years of age, were studied . Thirty-six adults sera were also included as controls . Polysaccharide of Hib was conjugated with tyramine and used to coat polystyrene microtitration plates . The determination of antibodies was performed by ELISA and the results are expressed in optical density units . The mean level of antibodies present in 6 sera from a patient with Bruton syndrome (17 uu), just before degammaglobulinotherapy, was considered as the limit of protection . RESULTS: The antibody titer correlated with age (p < 0.001) . The levels in children younger than 3 years (25.7 +/- 32.5) were lower than in older children (p < 0.001) . Of the children younger than 3 years of age, 53% did not reach the protective level of antibodies, compared to 15.2% of those between 3-6 years and 22.7% of children between 6 and 14 years . Whereas the mean antibody level increased with age in positive sera, the percentage of seropositive children decreased after 3 years of age . CONCLUSIONS: The number of children spontaneously protected against Hib is low (47%) until 3 years of age . Nevertheless, this number increases rapidly after this age . Immunization must be restricted to this age group.

Jpn J Antibiot, 1996 Nov, 49(11), 1013 - 23
{A clinical evaluation of azithromycin in the treatment of pediatric infection}; Furukawa S et al.; Fine granule preparation of azithromycin (AZM), a new macrolide antibiotic, was given to treat various infections in pediatric patients . Efficacies of AZM in a total of 21 patients (tonsillitis in six, bronchitis in five, pneumonia in five, impetigo contagiosa in three, staphylococcal scalded skin syndrome in one and bacterial enterogastritis in one) were rated "excellent" in 11 patients and "good" in eight . The remaining two cases were not included in the evaluation . AZM eradicated all strains of infection-causative bacteria identified in the 21 patients: Staphylococcus aureus in two, Streptococcus pneumoniae in four, Moraxella (Branhamella) catarrhalis in four, Haemophilus influenzae in six, Haemophilus parainfluenzae in three and Mycoplasma pneumoniae in one . One patient complained of mild diarrhea, while two patients showed increases in eosinophils as abnormal laboratory changes.

Jpn J Antibiot, 1996 Nov, 49(11), 1004 - 12
{Clinical study of a macrolide antibiotic, azithromycin, in pediatric patients}; Kobayashi Y et al.; Azithromycin (AZM), 10% fine granules or 100 mg capsules, were given orally to 27 children with various pediatric infections . The results of the study are shown below . 1 . Pharmacokinetic investigation . We studied plasma and urinary concentrations after 100 mg AZM capsules were given . One patient received 8.3 mg/kg of AZM once a day for 3 days, and AZM concentration in plasma was 0.033 microgram/ml 48 hours after the final dosing . Doses of 8.3 and 12.5 mg/kg body weight of AZM were respectively given to two patients once daily for 3 days . As a result, AZM concentrations in urine during a period between 96 and 120 hours post-dosing were 1.67 and 4.53 micrograms/ml, respectively, and urinary excretion rate in 120 hours after the first dosing was 10.54% in the patient that was given 12.5 mg/kg . 2 . Clinical investigation . Clinical efficacies were examined in 24 patients . Excellent results were obtained in 7 patients, good results in 14 patients, hence the clinical efficacy rate was 87.5% . Bacteriologically, Haemophilus influenzae strains isolates from 2 patients were eradicated in 1 and decreased in the other . Safety was evaluated in 26 patients . An adverse reaction was observed in 1 patient (urticaria) . Abnormal laboratory test results were observed in 2 patients, decreased WBC in 1 and elevation of eosinophils in the other . The above results suggest that AZM is a useful oral antibiotic for pediatric patients with infection with susceptible organisms.

Jpn J Antibiot, 1996 Nov, 49(11), 981 - 93
{Pharmacokinetic and clinical evaluation of azithromycin using fine granules or capsules in the pediatric patients}; Toyonaga Y et al.; Azithromycin (AZM), a new macrolide antibiotic, in fine granules and in capsules was studied for pharmacokinetic and clinical evaluations . 1 . Antibacterial activities . MIC profile of AZM was as follows: 0.78 approximately 1.56 micrograms/ml against Staphylococcus aureus, < or = 0.025 approximately 0.10 microgram/ml against Streptococcus pyogenes, 0.10 approximately 0.39 and 6.25 micrograms/ml against Streptococcus pneumoniae, < or = 0.025 approximately 0.39 microgram/ml against Moraxella(Branhamella) catarrhalis, 0.39 approximately 3.13 micrograms/ml against Haemophilus influenzae, and 0.20 approximately 6.25 micrograms/ml against Haemophilus parainfluenzae . 2 . Absorption and excretion . The elimination half-life of AZM after its administration at 10 mg/kg/day for three days was 28.1 approximately 46.1 hours . The cumulative urinary excretion rate in the first 120 hours after start of treatment was 4.01 approximately 8.47% . 3 . Clinical evaluation . AZM was given to 76 pediatric patients to treat following infections: pharyngitis in seven, tonsillitis in 11, bronchitis in 11, pneumonia in 19, Mycoplasma pneumonia in eight, scarlet fever in 13, infective enteritis in one, SSTI in four, and otitis media in two . Effectiveness of AZM was assessed in 75 patients and the drug was rated "excellent" or "good" in 71 resulting in an efficacy rate of 94.7%, 87.0% of the 46 cases indicated that AZM had eradicated bacteria identified before the treatment . One patient complained of moderate diarrhea which disappeared after treatment of anti-diarrheic . Abnormal laboratory changes were reported in 12 patients in the following: decreased leukocytes in eight, increased eosinophils in two, increased platelet count in one, and increased GPT in one . All cases of abnormality was deemed mild in severity and clinically insignificant.

Rev Prat, 1996 Nov 1, 46(17), 2099 - 103
{Antibiotic strategy in lower respiratory infections in children}; Gendrel D; Antibiotic treatment of community-acquired lower respiratory tract infections in children remains widely empiric since the determination of the causative agent is difficult and rarely done . Antibiotic therapy is not necessary in all cases since viral infections are frequent . Antibiotic choice must take in account the possibility of pneumococcal infections . Pneumococci are not the most frequent causative agent but always provoke severe infections with a risk of death and amoxicillin is the first-line choice, with high doses (100 mg/kg/d) because of the emergence of strains with a reduced sensitivity to penicillin . In younger children, Haemophilus influenzae infections are frequent and clavulanic acid must be associated . When intravenous route is required, 3rd generation cephalosporins are useful . The clinical failure of beta-lactams antibiotics is widely due, after the age of two years, to a Mycoplasma pneumoniae infection and a macrolide is required as second-line choice . Markers specific of viral infection are needed in children to reduce antibiotic prescription.

Rev Prat, 1996 Nov 1, 46(17), 2056 - 61
{Epidemiology of acute bronchopulmonary infections in children}; Garcia J; In infants and young children acute lower respiratory infection is the most common cause of morbidity and death especially in developing countries . Factors that contribute to the increased susceptibility to respiratory pathogens include young age, season, sex, indoor pollution, large family size, malnutrition, low immunocompetence, socioeconomic disadvantage . The epidemiology of acute respiratory infections in childhood seems similar worldwide . In all countries, respiratory syncytial virus, parainfluenzae virus 1 and 3 influenzae A and B viruses and adenovirus are reported to be the main causes of acute respiratory infections . Six microorganisms are responsible of 90% of documented acute bacterial pulmonary infections, Streptococcus pneumoniae, Mycoplasma pneumoniae, Chlamydia pneumoniae, Chlamydia trachomatis, Haemophilus influenzae, Staphylococcus . Mixed viral and bacterial infections occur frequently (30%) . The role of respiratory viruses in predisposing to colonization and invasion of bacterial organisms has often been suggested . In recent years acquired resistance against antibiotic for H . influenzae and S . pneumoniae has emerged.

Microbiology, 1996 Nov, 142 ( Pt 11), 3047 - 56
The 25 degrees-36 degrees region of the Bacillus subtilis chromosome: determination of the sequence of a 146 kb segment and identification of 113 genes; Yamane K et al.; We determined a 146 kb contiguous sequence at the 25 degrees-36 degrees region of the Bacillus subtilis chromosome containing the amyE-srfA segment . Among the 113 ORFs identified, 33 are already known . functions were assigned to 38 ORFs by a search of non-redundant protein sequence data banks and those of 16 ORFs were suggested through significant similarity with reported sequences . The amino acid sequences of 13 of the ORfs were similar to proteins of unknown function of Escherichia coli, Haemophilus influenzae and other species . We did not find similarities for 29 ORFs to any known proteins . The 146 kb region is rich in enzymes (35 ORFs) related to the metabolism of low molecular mass compounds and five genes for surfactin production occupy about 26 kb of the region.

J Antimicrob Chemother, 1996 Nov, 38(5), 881 - 4
Clinical features of patients with beta-lactamase producing Haemophilus influenzae isolated from sputum; Johnson SR et al.; Ampicillin resistance amongst isolates of Haemophilus influenzae is of increasing concern but its clinical impact is unclear . We performed a retrospective study of 34 hospital patients with lower respiratory tract infection (LRTI) caused by beta-lactamase positive (beta+) H . influenzae and compared these with 34 control patients with LRTI caused by beta-lactamase negative (beta-) strains . Chronic obstructive pulmonary disease was the most common underlying condition in both groups (beta+ 23/24, beta- 25/34) . A recent course of antibiotics, especially ampicillin/amoxycillin, was significantly (P < 0.05) more common in the beta+ group (beta+ 17/34, beta- 3/34) . Other pathogens were isolated more frequently in the beta+ group (beta+ 10/34, beta- 4/34) . There was no difference in outcome between the two groups.

J Antimicrob Chemother, 1996 Nov, 38(5), 849 - 52
Comparison of the Etest with agar dilution for antimicrobial susceptibility testing of Haemophilus ducreyi; Lagergard T et al.; Two methods for determining the sensitivity of Haemophilus ducreyi to antimicrobial agents were compared; an agar dilution method and the Etest . The MICs of seven antimicrobial agents; penicillin V, ampicillin, trimethoprim, tetracycline, chloramphenicol, erythromycin and ciprofloxacin were determined for 20 strains of H . ducreyi . The MICs determined with the Etest and with the agar dilution method, showed 86% agreement within one two-fold dilution . The Etest is a good alternative method to the agar dilution technique for antimicrobial susceptibility testing of H . ducreyi . Owing to its simplicity and good reproducibility, the test is well suited for routine use in clinical microbiology laboratories in developing countries.

J Antimicrob Chemother, 1996 Nov, 38(5), 777 - 86
Antimicrobial susceptibility of Haemophilus influenzae, Neisseria meningitidis and Streptococcus pneumoniae isolates causing meningitis in The Netherlands, 1993-1994; Enting RH et al.; The increasing antimicrobial resistance among pathogens frequently isolated from patients with bacterial meningitis formed the rationale to perform a surveillance study to determine the prevalence of resistance in The Netherlands . Haemophilus influenzae strains (n = 316) isolated from cerebrospinal fluid (CSF), 1125 meningococcal strains isolated from blood or CSF and 398 pneumococcal strains isolated from CSF in 1993 and 1994 were tested by the Etest for susceptibility to commonly prescribed antibiotics for the treatment of community-acquired meningitis . In H . influenzae strains ampicillin-resistance occurred in 7.0%, resistance to chloramphenicol in 2.2%, and resistance to both antibiotics in 0.9% . The prevalence of intermediate penicillin-resistance in meningococci was 3.3% . Resistance to rifampicin was rarely found (0.1%) . Intermediate penicillin-resistance in pneumococci was found in only 0.5% of isolates . All 1839 isolates were susceptible to ceftriaxone . Based on these results, we conclude that empirical therapy of childhood community-acquired bacterial meningitis with amoxycillin and chloramphenicol is no longer justified in children who have not been vaccinated against H . influenzae type b . In vaccinated or older children and adults, amoxycillin is a rational choice for empirical treatment of meningitis . The prophylactic use of rifampicin in contacts of patients with meningococcal disease is still applicable.

Chemotherapy, 1996 Nov-Dec, 42(6), 465 - 7
Effect of phagocytosis of bacteria on the uptake of sparfloxacin by human neutrophils; Garcia I et al.; The effect of phagocytosis of Escherichia coli, Staphylococcus aureus and Haemophilus influenzae on the intracellular penetration of sparfloxacin into human polymorphonuclear neutrophils (PMN) was evaluated . The intracellular concentration of sparfloxacin was similar in PMN containing intracellular bacteria and in resting PMN {cellular to extracellular ratio (C/ E) > 8.0} . The uptake of sparfloxacin by human PMN significantly decreased during phagocytosis of opsonized H . influenzae but still reached intracellular concentrations five times greater than the extracellular ones . This effect was not observed with E . coli and S . aureus.

Clin Nephrol, 1996 Nov, 46(5), 287 - 95
Role of IgA, IgG, and IgM antibodies against Haemophilus parainfluenzae antigens in IgA nephropathy; Suzuki S et al.; We have recently demonstrated glomerular deposition of outer membranes of Haemophilus parainfluenzae (HP) antigens (OMHP) and the presence of IgA antibody against OMHP in patients with IgA nephropathy (IgA-N) . In this study, we analyzed IgA-, IgG-, and IgM-classes of antibodies against OMHP, and the relationship between these antibodies and renal lesions in IgA-N . The subjects included 44 patients with IgA-N and 62 patients with outer glomerular diseases (OGD); the latter group consisted of 23 patients with predominantly IgG or IgM deposits and small amounts of IgA in the mesangium (group A), and 39 with IgG or IgM deposits without IgA (group B) . IgA, IgG, and IgM antibodies against OMHP in patients sera were detected by enzyme-linked immunosorbent assay (ELISA) . Immunoblotting demonstrated that the IgA, IgG, and IgM antibodies against OMHP in the sera of IgA-N patients bound to components of OMHP with molecular weights of 19.5, 30, and 40.5 kD . The amino acid compositions of these three OMHP components were similar to those reported for the outer membrane protein (OMP) P6 precursor, OMP P5, and OMP P2 (porin) of Haemophilus influenzae . Both IgA-N and group A patients, (i.e . those with IgA-related renal disease), demonstrated a significantly higher level of IgA antibodies against OMHP than did group B patients . However, only IgA-N patients revealed a significant correlation between the IgA-antibody titer and degree of glomerular changes . IgA-N patients with macroscopic hematuria or arterio(lo)sclerosis had a significantly higher IgA antibody titer than other IgA-N patients . There was no relationship between renal lesions and IgG or IgM antibody titers in any group . These findings suggest that IgA antibodies against OMHP are significantly increased in patients with IgA-related renal disease compared to those without mesangial IgA deposits and that a significant relationship between these antibodies and renal lesions exists only in patients with IgA-N.

Arch Pediatr, 1996 Nov, 3(11), 1084 - 90
{Purulent pericarditis in children: a 15 year-experience}; Thebaud B et al.; BACKGROUND: Purulent pericarditis is rare in developed countries, but its prognosis is regarded as serious . Early diagnosis and appropriate treatment should prevent complications . PATIENTS: Among the 119 pericarditis without congenital cardiopathy admitted in our pediatric cardiology unit between 1979 and 1994, 19 were purulent . The mean age of these 13 boys and six girls was 3 years (range: 3 months to 10 years) . Symptoms always pointed to a chest disease whether pericarditis occurred first (n = 13) or it complicated evolution of a known infectious process (n = 6) . Tamponnade was present in seven infants upon admission and required urgent pericardiocentesis or drainage . An extrapericardial infectious site was found in 11 cases: six pulmonary infections, three osteomyelitis, one cellulitis and one sinusitis . An organism was isolated in 17 cases, 14 times in the pericardial fluid, eight times on blood culture . The identified bacteria were: Staphylococcus aureus (n = 6), Haemophilus influenzae (n = 4), Streptococcus A (n = 3), Streptococcus pneumoniae (n = 3), Meningococcus (n = 1) . Treatment consisted of intra-venous antibiotics associated 15 times to surgical drainage of the pericardium . One infant had no pericardiocentesis and no drainage because he presented late with constrictive pericarditis and needed pericardectomy . All infants healed but four developed contrictive pericarditis and required pericardectomy; none of these four patients had early drainage (two had no drainage at all) . Pericardectomy, carried out between 2 and 6 months after the beginning of pericarditis with adiastolic signs and pericardial thickening, permitted healing in all cases and disappearance of all cardiac symptoms . CONCLUSIONS: Early diagnosis and treatment of purulent pericarditis, especially early pericardial drainage, are the best ways of avoiding constriction.

Am J Trop Med Hyg, 1996 Nov, 55(5), 516 - 20
The immunogenicity of three Haemophilus influenzae type B conjugate vaccines after a primary vaccination series in Philippine infants; Capeding MR et al.; Serum antibody responses to three Haemophilus influenzae type b (Hib) capsular polysaccharide-protein conjugate vaccine (PRP-OMP, PRP-T, and HbOC) were evaluated in 174 Philippine infants after a primary vaccination series . Children were randomized to receive one of the Hib vaccines (Hib groups) or into a control group . Vaccination was carried out at six, 10 and 14 weeks of age based on the local Expanded Program of Immunization schedule . Sera were collected at six weeks of age for the Hib groups and one month after the third dose for all subjects . Anti-Hib concentrations were determined by the Farr-type radioimmunoassay . There were no significant differences (P = 0.3626) in the prevaccination anti-Hib geometric mean concentration (GMC) among the three Hib groups . Differences in the GMC after the primary series of three doses were significant (P < 0.0001); GMC was highest for PRP-T (6.62 micrograms/ml), followed by HbOC (1.9 micrograms/ml), then PRP-OMP (1.06 micrograms/ml), and lowest for the control group (0.11 microgram/ml) . We conclude that all three Hib conjugate vaccines (PRP-T, HbOC, and PRP-OMP) were immunogenic after three primary doses among Philippine infants.

Mol Microbiol, 1996 Nov, 22(3), 497 - 507
PhhR, a divergently transcribed activator of the phenylalanine hydroxylase gene cluster of Pseudomonas aeruginosa; Song J et al.; Pseudomonas aeruginosa was recently found to possess a cluster of structural genes encoding phenylalanine hydroxylase (PhhA), carbinolamine dehydratase (PhhB), and aromatic aminotransferase (PhhC) . We now report the presence, in the flanking upstream region, of a divergently transcribed gene (phhR) encoding an activator protein . Inactivation of phhR markedly reduced expression of the structural genes . PhhR belongs to the large prokaryote family of sigma 54 enhancer-binding proteins, and activation of the phh operon by PhhR in P . aeruginosa required rpoN . The closest homologues of PhhR are the TyrR proteins from Escherichia coli and Haemophilus influenzae . E . coli TyrR is an unusual member of the homologue family in that the transcriptional units regulated by tyrR are driven by sigma 70 promoters . P . aeruginosa phhR was able to replace E . coli tyrR as a repressor of the aroF-tyrA operon (but not as an activator of mtr) in the heterologous E . coli system . Two regions that resemble E . coli TyrR boxes were identified in the intervening region between phhR and phhA . We propose that one or both boxes may be the target of PhhR acting as an autogenous repressor at a sigma 70 promoter in one direction . In the other direction, one or both boxes may be the upstream activator sequence targeted by PhhR to facilitate expression of the phh operon from a sigma 54 promoter . The phh operon was strongly induced in fructose- or glucose-based minimal medium by L-phenylalanine . Inactivation of phhR in P . aeruginosa abolished ability to utilize either L-phenylalanine or L-tyrosine as a sole source of carbon for growth.

Mol Microbiol, 1996 Nov, 22(3), 437 - 47
A third family of allelic hsd genes in Salmonella enterica: sequence comparisons with related proteins identify conserved regions implicated in restriction of DNA; Titheradge AJ et al.; Salmonella enterica serovar blegdam has a restriction and modification system encoded by genes linked to serB . We have cloned these genes, putative alleles of the hsd locus of Escherichia coli K-12, and confirmed by the sequence similarities of flanking DNA that the hsd genes of S . enterica serovar blegdam have the same chromosomal location as those of E . coli K-12 and Salmonella enterica serovar typhimurium LT2 . There is, however, no obvious similarity in their nucleotide sequences, and while the gene order in S . enterica serovar blegdam is serB hsdM, S and R, that in E . coli K-12 and S . enterica serovar typhimurium LT2 is serB hsdR, M and S . The hsd genes of S . enterica serovar blegdam identify a third family of serB-linked hsd genes (type ID) . The polypeptide sequence predicted from the three hsd genes show some similarities (18-50% identity) with the polypeptides of known and putative type I restriction and modification systems; the highest levels of identity are with sequences of Haemophilus influenzae Rd . The HsdM polypeptide has the motifs characteristic of adenine methyltransferases . Comparisons of the HsdR sequence with those for three other families of type I systems and three putative HsdR polypeptides identify two highly conserved regions in addition to the seven proposed DEAD-box motifs.

Microb Pathog, 1996 Nov, 21(5), 343 - 56
Selective adherence of non-typeable Haemophilus influenzae (NTHi) to mucus or epithelial cells in the chinchilla eustachian tube and middle ear; Miyamoto N et al.; Frozen sections of chinchilla Eustachian tube (ET) and middle ear mucosa were incubated with either FITC-labeled non-typeable Haemophilus influenzae (NTHi) or Bordetella pertussis . The number of bacteria adherent to "roof" vs "floor" regions was compared for each of three anatomic portions of the ET and for middle ear epithelium noting whether bacteria adhered to mucus or to epithelial cells . NTHi strains adhered significantly greater to mucus in the ET lumen whereas B . pertussis preferentially adhered to epithelial cells lining the ET (P < or = 0.05) . A non-fimbriated isogenic mutant of NTHi adhered significantly less to mucus than the parental isolate at all sites of the ET floor (P < or = 0.05) . Isolated fimbrin protein adhered to ET mucus and blocked adherence of whole organisms . Treatment with the mucolytic agent N-acetyl-L-cysteine resulted in significantly reduced adherence of NTHi to mucus (P < or = 0.001) and eliminated the ability to detect binding of isolated fimbrin protein . N-acetyl-L-cysteine treatment did not affect adherence of either B . pertussis or NTHi to epithelial cells . These data indicated that NTHi may mediate ascension of the ET from the nasopharynx primarily via adherence to and growth in mucus overlying the floor region of the tubal lumen . The OMP P5-homologous fimbriae were shown to contribute to this binding.

Bone Marrow Transplant, 1996 Nov, 18(5), 961 - 7
A comparison of early and late vaccination with Haemophilus influenzae type b conjugate and pneumococcal polysaccharide vaccines after allogeneic BMT; Parkkali T et al.; Forty-five adult allogeneic BMT recipients were randomized to receive Haemophilus influenzae type b (Hib)-diphtheria toxoid conjugate vaccine (PRP-D) at 6 and pneumococcal polysaccharide vaccine (Pnc PS) at 8 months (early group; n = 23) or at 18 and 20 months (late group; n = 22) after BMT, respectively . Serum antibody concentrations against Hib and Pnc PS types with varying immunogenicity (serotypes 3, 6B, and 19F) were measured by ELISA . The responses at 1 month after vaccination with PRP-D and Pnc PS were poor and similar in the two vaccination groups, except that two-fold responses in the concentration of antibodies to the most immunogenic Pnc serotype 3 occurred more frequently in the late group . In the late group recipients, the antibody responses elicited by Pnc serotypes 3 and 19F correlated with the concentrations of the corresponding antibodies in the donor . This study on allogeneic BMT recipients shows that vaccination at 6-8 months after BMT with one dose of PRP-D and Pnc PS vaccine is as immunogenic as vaccination at 18-20 months after BMT, and suggests that donor immunity to Pnc PS affects recipient responses to Pnc PS vaccination . Consequently, as among allogeneic BMT recipients the greatest risk for infections by encapsulated bacteria occurs during the first 1-2 years after BMT, Hib and Pnc vaccines should not be given later than 6-8 months post-BMT.

Nucleic Acids Res, 1996 Nov 1, 24(21), 4263 - 72
Frequent oligonucleotides and peptides of the Haemophilus influenzae genome; Karlin S et al.; The complete Haemophilus influenzae genome (1.83 Mb, Rd strain) provides opportunities for characterizing global genomic inhomogeneities and for detecting important sequence signals . Along these lines, new methods for identifying frequent words (oligonucleotides and/or peptides) and their distributions are applied to the H.influenzae genome with some comparisons and contrasts made with frequent words of other bacterial genomes . Three major classes of frequent oligonucleotides stand out: (i) oligos related to the familiar uptake signal sequences (USSs), AAGTGCGGT (USS+) and its inverted complement (USS-), (ii) multiple tetranucleotide iterations and (iii) intergenic dyad sequences (ISDs) found as AAGCCCACCCTAC and its dyad form . The USS+ and USS- occur in almost equal counts, are remarkably evenly spaced around the genome, and appear predominantly in the same reading frame of protein coding domains (USS+ translated to Ser-Ala-Val, USS- translated to Thr-Ala-Leu) . These observations suggest that USSs contribute to global genomic functions, for example, in replication and/or repair processes, or as membrane attachment sites, or as sequences helping to pack DNA . The long tetranucleotide iterations, virtually unique to H.influenzae (i.e., unknown in other prokaryotes), through polymerase slippage during replication and/or homologous recombination may produce subpopulations expressing alternative proteins . The 13 bp frequent IDS words, invariably intergenic, occur mostly in clusters and provide potential for complex secondary structures suggesting that these sequences may be important signals for regulating the activity of their flanking genes . The frequent oligopeptides of H.influenzae are principally of two kinds--those induced by oligonucleotide frequent words (USSs, tetranucleotide iterations), and those associated with ATP or GTP binding sites that are generally composed of three motifs: the A-box which contributes to delineating the binding pocket; the B-box which functions in hydrolysis; and the C-box whose function is unknown . The A-box occurs fairly universally in prokaryotes and eukaryotes . The B- and C-motifs appear to be specialized to various functional groups (e.g., transport, recombination, chaperone activity) . Other putative motifs correspond to homologs of Escherichia coli motifs, for example, are associated with proteins of transcriptional processing, aminoacyl-tRNA synthetases and proteins functioning in electron transfer.

Nucleic Acids Res, 1996 Nov 1, 24(21), 4146 - 51
Statistical evidence for a biochemical pathway of natural, sequence-targeted G/C to C/G transversion mutagenesis in Haemophilus influenzae Rd; Merkl R et al.; Markov chain analysis of the Haemophilus influenzae Rd genome reveals striking under-representation of three palindromic tetranucleotide strings (CCGG, GGCC and CATG), accompanied by over-representation of six tetranucleotide strings that are derived from the former by exchanging strand location of the two residues making up a G/C nucleotide pair at the terminal palindrome position . Constraints are outlined for a molecular model able to explain the phenomenon as the result of sequence-targeted, enzyme-driven G/C to C/G transversion mutagenesis . Possible participation in the process by components of known DNA mismatch repair or restriction/modification systems (in particular, cytosine methylation) is discussed . The effect widens the spectrum of enzyme-driven, specific mutagenesis beyond the formerly described C/G to T/A transition (VSP repair of Escherichia coli) . Potential evolutionary benefits of enzymatic pathways of specific mutagenesis can be envisioned.

J Bacteriol, 1996 Nov, 178(22), 6564 - 70
Duplication of pilus gene complexes of Haemophilus influenzae biogroup aegyptius; Read TD et al.; Brazilian purpuric fever (BPF) is a recently described pediatric septicemia caused by a strain of Haemophilus influenzae biogroup aegyptius . The pilus specified by this bacterium may be important in BPF pathogenesis, enhancing attachment to host tissue . Here, we report the cloning of two haf (for H . influenzae biogroup aegyptius fimbriae) gene clusters from a cosmid library of strain F3031 . We sequenced a 6.8-kb segment of the haf1 cluster and identified five genes (hafA to hafE) . The predicted protein products, HafA to HafD, are 72, 95, 98, and 90% similar, respectively, to HifA to HifD of the closely related H . influenzae type b pilus . Strikingly, the putative pilus adhesion, HifE, shares only 44% identity with HafE, suggesting that the proteins may differ in receptor specificity . Insertion of a mini-gammadelta transposon in the hafE gene eliminated hemadsorption . The nucleotide sequences of the haf1 and haf2 clusters are more than 99% identical . Using the recently published sequence of the H . influenzae Rd genome, we determined that the haf1 complex lies at a unique position in the chromosome between the pmbA gene and a hypothetical open reading frame, HI1153 . The location of the haf2 cluster, inserted between the purE and pepN genes, is analogous to the hif genes on H . influenzae type b . BPF fimbrial phase switching appears to involve slip-strand mispairing of repeated dinucleotides in the pilus promoter . The BPF-associated H . influenzae biogroup aegyptius pilus system generally resembles other H . influenzae, but the possession of a second fimbrial gene cluster, which appears to have arisen by a recent duplication event, and the novel sequence of the HafE adhesin may be significant in the unusual pathogenesis of BPF.

Clin Infect Dis, 1996 Nov, 23(5), 1081 - 90
High rate of nasopharyngeal carriage of potential pathogens among children in Greenland: results of a clinical survey of middle-ear disease; Homoe P et al.; Nasopharyngeal and middle-ear colonization with bacteria and viruses, including Mycoplasma pneumoniae and chlamydiae, was investigated in a survey of 54 children with acute otitis media (AOM) and 201 control children without AOM in Greenland . In total, 98% with AOM and 91% without AOM carried potentially pathogenic bacteria in the nasopharynx . Two or more potentially pathogenic species were carried by 78% with AOM and 57% without AOM . Haemophilus influenzae was found in 92% and 77%, respectively, but only Streptococcus pneumoniae was found significantly more often in the nasopharynx of children with AOM than in age-matched controls (P < .03) . The two species were found in 22 of 24 ear-discharge specimens . Nine children (three with AOM) had chlamydiae in the nasopharynx, and seven of them reported rhinitis . Enteroviruses or rhinoviruses were detected in 23 nasopharyngeal specimens from 39 children with AOM, in 13 such specimens from 39 children without AOM (P = .040), and in 4 of 14 ear-discharge specimens . The potentially pathogenic load in the nasopharynx was massive, suggesting an association with the high prevalence of otitis media among children in Greenland.

Clin Infect Dis, 1996 Nov, 23(5), 1069 - 74
Resistance patterns of Streptococcus pneumoniae and Haemophilus influenzae isolates recovered in Egypt from children with pneumonia . The Antimicrobial Resistance Surveillance Study Group; Ostroff SM et al.; Treatment of childhood pneumonia in developing countries requires knowledge of susceptibility patterns for Streptococcus pneumoniae and Haemophilus influenzae . Between October 1991 and April 1993, a surveillance survey of antimicrobial resistance was performed at two fever hospitals in Egypt; nasopharyngeal swab and blood specimens obtained from 1,635 children with pneumonia were cultured for these organisms . Susceptibility testing of these organisms was performed . At least one of these organisms was isolated from nasopharyngeal swab specimens from 73% of the children; 3.7% of blood cultures were positive . For S . pneumoniae strains, 70.9% of nasopharyngeal isolates were calculated to be susceptible to penicillin vs . 77.6% of blood isolates; the percentages of isolates susceptible to co-trimoxazole were 73.0% and 75.0%, respectively . For H . influenzae strains, 93.0% of nasopharyngeal isolates were calculated to be susceptible to ampicillin vs . 100% of blood isolates; the percentages of isolates susceptible to co-trimoxazole were 84.9% and 100%, respectively . Although most S . pneumoniae and H . influenzae strains associated with childhood pneumonia in Cairo were susceptible to penicillins and co-trimoxazole, antimicrobial resistance did not occur.

J Med Microbiol, 1996 Nov, 45(5), 380 - 2
Inhibition of growth of fungi pathogenic to man by Stenotrophomonas maltophilia; Kerr JR; Ten strains of Stenotrophomonas maltophilia were tested by a cross-streaking method for in-vitro inhibition of 10 strains of fungi known to infect man; these were Candida krusei, C . keyfr, C . guillermondii, C . tropicalis, C . lusitaniae, C . parapsilosis, C . pseudotropicalis, C . albicans, C . glabrata and Aspergillus fumigatus . Significant inhibition of fungal growth was observed with all 10 strains of S . maltophilia, but not with Escherichia coli NCTC 10418, Staphylococcus aureus NCTC 6571 and Haemophilus influenzae NCTC 11931.

J Med Microbiol, 1996 Nov, 45(5), 372 - 5
Microbiology of the transition from acute to chronic maxillary sinusitis; Brook I et al.; Repeated aspirations of sinus secretions by endoscopy was performed in five patients over a period of 34-50 days and, ultimately, surgical drainage was done in three who presented with acute maxillary sinusitis that did not respond to antimicrobial therapy and became chronic . The aspirates were cultured for aerobic and anaerobic bacteria . Most of the bacteria isolated from the first culture were aerobic or facultative bacteria: Streptococcus pneumoniae (three isolates), Haemophilus influenzae non-type-b (two) and Moraxella catarrhalis (one) . Three of these cultures yielded bacteria that were resistant to the antimicrobial agents prescribed for treatment . Failure to respond to therapy was associated with the emergence of resistant aerobic and anaerobic bacteria in subsequent aspirates . These organisms included Fusobacterium nucleatum, pigmented Prevotella and Porphyromonas spp . and Peptostreptococcus spp . Eradication of the infection was achieved in all instances following the administration of antimicrobial agents effective against these bacteria, and in three instances by surgical drainage . This study illustrates the microbial dynamics of maxillary sinusitis that did not respond to antimicrobial therapy.

Pediatrics, 1996 Nov, 98(5), 898 - 904
Interchangeability of Haemophilus influenzae type b vaccines in the primary series: evaluation of a two-dose mixed regimen; Bewley KM et al.; OBJECTIVE: To evaluate two- or three-dose "mixed" regimens of Haemophilus influenzae type b conjugate vaccines in the priming series . DESIGN: Two randomized clinical trials with 140 and 181 infants, respectively . SETTING: Private practices in New Orleans and Chicago . METHODS: In trial I, infants received one of four regimens . Two were recommended regimens for polyribosylribitol phosphate (PRP)-meningococcal protein conjugate (M) and PRP-tetanus toxoid conjugate (T) . Two mixed regimens consisted of M at 2 months followed by two doses of T or PRP-diphtheria toxoid conjugate (D) at 4 and 6 months . Trial II consisted of three groups . Two were recommended regimens for M and T . The third was a two-dose mixed regimen consisting of M at 2 months and T at 4 months . Parents were interviewed and instructed to record side effects after each vaccination . Serum was assayed for H influenzae type b anticapsular antibody (anti-PRP) . RESULTS: Minor differences in safety profiles likely reflected alpha error . In trial I, M (lot 0884T, one of several known to have had decreased immunogenicity), probably primed for substantial increase in serum antibody when D or T was given at 4 and 6 months . In trial II, infants who received the two-dose mixed regimen (M from immunogenic lot 0116W at 2 months and T at 4 months) had a significantly higher mean area under the curve than recipients of the three-dose TTT regimen when antibody concentration was plotted against age, although the geometric mean anti-PRP antibody concentration for the MT-recipients was significantly lower at 7 months . CONCLUSIONS: M used in trial I may have primed infants despite poor immunogenicity . The two-dose mixed regimen (MT-) in trial II produced a mean anti-PRP antibody concentration with higher sustained anti-PRP concentrations from 2 to 7 months, as judged by the area under the curve, but a lower mean anti-PRP antibody concentration at 7 months.

J Clin Microbiol, 1996 Nov, 34(11), 2770 - 7
On the nature and use of randomly amplified DNA from Staphylococcus aureus; van Leeuwen W et al.; Various DNA-based methods have been introduced to genetically type Staphylococcus aureus strains but not a single technique is universally applicable . In order to search for DNA probes suitable for differentiating strains, randomly amplified polymorphic DNA patterns were generated for 243 S . aureus strains and a single isolate of Staphylococcus intermedius . All fingerprints were examined for unique amplicons, and the nature of 42 of these DNA fragments was investigated . Partial DNA sequences were determined, and several homologies were discovered with known S . aureus sequences (plasmid pSH6 DNA with insertion sequences, agrA and agrB sequences, hld genes, the gene for 23S rRNA, the lysyl tRNA synthetase gene, and the threonyl tRNA synthetase gene) and with genes from other species (Haemophilus influenzae bexA and Bacillus subtilis spoF and ctrA) . Thirty fragments were of previously unknown origin . In Southern blots containing Eco RI-digested DNA from S . aureus strains and the S . intermedius strain, nine probes demonstrated the capacity to differentiate strains on the basis of the presence or absence of the sequence element in the staphylococcal genome involved . The remainder of the probes displayed restriction fragment length polymorphisms (n = 12), hybridized in a homogeneously positive fashion (n = 13) or hybridized only with their source strains (n = 8) (four of the latter were specific to S . intermedius) . Three of the nine strain-specific probes were overlapping, and two of the others were found to display a high level of inconsistency among epidemiologically related strains . Thus, five strain-specific probes remained that, in a 5-digit typing system, accurately distinguished epidemiologically related and unrelated strains of S . aureus . We conclude that application of strain-specific DNA probes, selected on the basis of differing randomly amplified polymorphic DNA patterns, promises to become a technically simple, robust, and reproducible tool that may significantly facilitate the study of the epidemiology of S . aureus infections.

J Clin Microbiol, 1996 Nov, 34(11), 2760 - 5
Clonality of multidrug-resistant nontypeable strains of Haemophilus influenzae; Fuste MC et al.; The genetic structure of a population of multidrug-resistant nontypeable (unencapsulated) Haemophilus influenzae strains isolated at a hospital in Barcelona, Spain, was investigated by using multilocus enzyme electrophoresis to determine the allelic variation in 15 structural loci . In our study we have also included some antimicrobial agent-susceptible strains isolated at the same hospital . All enzymes were polymorphic for two to eight electromorphs, and the analysis revealed 43 distinct electrophoretic types among the 44 isolates . The mean genetic diversity of the entire population was 0.55 . Multilocus linkage disequilibrium analysis of the isolates revealed a strong association between alleles, suggesting little possibility of recombination . Furthermore, the dendrogram and the allele mismatch distribution are typical of a population with no extensive genetic mixing.

J Infect Dis, 1996 Nov, 174 Suppl 3, S302 - 5
Analysis of Haemophilus influenzae type B conjugate and diphtheria-tetanus-pertussis combination vaccines; Eskola J; Combined vaccines consisting of diphtheria (D), tetanus (T), whole cell pertussis (Pw), and Haemophilus influenzae type b (Hib) conjugates are safe and immunogenic; however, there has been a tendency toward lower antibody responses in persons receiving mixtures of DTPw and Hib vaccines . Possible explanations are physical or chemical incompatibilities or immunologic interference . Mixtures of DT-acellular pertussis (Pa) and Hib conjugates have been evaluated as booster vaccinations in children, and preliminary data indicate good safety and acceptable immunogenicity . A study evaluating the combined administration of DTPa, Hib conjugate, and inactivated polio vaccine in infants is ongoing in Finland . Preliminary data suggest that the combination is safe . When the first serum samples were assayed, no major differences in pertussis, diphtheria, and tetanus antibodies were found among children given the vaccines mixed or separately . Hib and polio antibody results are not yet available.

J Neurosurg, 1996 Nov, 85(5), 949 - 52
Septic cavernous sinus thrombosis following transsphenoidal craniotomy . Case report; Sadun F et al.; The authors present a case of late-onset cavernous sinus thrombosis in a 74-year-old man who had undergone transsphenoidal craniotomy for a pituitary macroadenoma 9 weeks previously . The patient developed headache, rapidly progressive ophthalmoplegia, and signs of orbital congestion . After 2 days of ineffective broad spectrum antibiotic therapy he underwent a second transsphenoidal craniotomy for abscess drainage . Intraoperative cultures grew 4+ non-hemolytic Streptococcus, 4+ Staphylococcus coagulase negative, and 4+ Haemophilus influenzae . The patient was maintained on intravenous antibiotic therapy for the following 6 weeks, resulting in a complete clinical recovery . To the authors' knowledge, this is the first report of a septic cavernous sinus thrombosis following a transsphenoidal craniotomy.

J Bacteriol, 1996 Nov, 178(21), 6366 - 8
Role of the two-component signal transduction and the phosphoenolpyruvate: carbohydrate phosphotransferase systems in competence development of Haemophilus influenzae Rd; Gwinn ML et al.; Haemophilus influenzae Rd becomes competent for transformation by nutritional downshift or transient anaerobic growth through a process that requires cyclic AMP receptor protein and adenylate cyclase . Insertion mutations in crr or ptsI of the phosphoenolpyruvate:carbohydrate phosphotransferase system lowered transformation frequencies, and the effect was reversed by the addition of cyclic AMP . However, insertions into H . influenzae homologs of two-component signal transduction genes had no effect on competence.

J Bacteriol, 1996 Nov, 178(21), 6281 - 7
Characterization of the genetic locus encoding Haemophilus influenzae type b surface fibrils; St Geme JW 3rd et al.; Haemophilus influenzae is a common gram-negative pathogen that initiates infection by colonizing the upper respiratory tract epithelium . In previous work, we reported the isolation of a locus involved in expression of short, thin surface fibrils by H . influenzae type b and presented evidence that surface fibrils promote attachment to human epithelial cells . In the present study, we determined that the fibril locus is composed of one long open reading frame, designated hsf, which encodes a protein (Hsf) with a molecular mass of approximately 240 kDa . The derived amino acid sequence of the hsf product demonstrated 81% similarity and 72% identity to a recently identified nontypeable H . influenzae adhesin referred to as Hia . In experiments with a panel of eight cultured cell lines, the Hsf and Hia proteins were found to confer the same binding specificities, suggesting that hsf and hia are alleles of the same locus . Southern analysis and mutagenesis studies reinforced this conclusion . Further investigation revealed that an hsf homolog is ubiquitous among encapsulated H . influenzae strains and is present in a subset of nontypeable Haemophilus strains as well . We speculate that the hsf gene product plays an important role in the process of respiratory tract colonization by H . influenzae.

J Immunol, 1996 Nov 1, 157(9), 4016 - 27
The progeny of a single virgin B cell predominates the human recall B cell response to the capsular polysaccharide of Haemophilus influenzae type b; Barington T et al.; Restricted V region diversity is a key feature of Abs to many haptens and simple polysaccharides . Two possible mechanisms exist: 1) selection of many clonally unrelated B cells using very similar or identical VDJ and VJ rearrangements; and 2) selection of a heavily expanded progeny of few virgin B cells . How many virgin B cells eventually give rise to the total Ab response to a simple Ag is a fundamental immunologic question . In this report, we address this question in human adults by analyzing the rearranged VkappaJkappa genes of B cells responding to a single dose of the capsular polysaccharide of Haemophilus influenzae type b coupled to tetanus toxoid . We combined affinity purification of circulating vaccine-induced Ab-secreting cells with PCR amplification of cDNA followed by cloning and sequencing . Forty-eight and 42 kappa VJ gene transcripts were analyzed from two adults, respectively . Both individuals used extremely restricted repertoires with >90% of the cells using a single Vkappa gene rearranged to a single Jkappa gene . Despite the fact that the Ab responses engaged high numbers of Ab-secreting cells, analysis of the many shared, somatically acquired mutations showed that the majority of the cells originated from a common virgin B cell . Kinetic considerations implied that an extremely selected population of hypermutated memory B cells must have existed in these individuals before the first systemic immunization with the Ag . A possible role for the mucosal immune system in the priming and selection of these cells is proposed.

Infect Immun, 1996 Nov, 64(11), 4834 - 7
Cloning, nucleotide sequence, and expression of the gene coding for a ribosome releasing factor-homologous protein of Brucella melitensis; Vizcaino N et al.; The gene coding for a Brucella melitensis cytosoluble protein (CP24) that is immunogenic in infected sheep and a major component of brucellin INRA was cloned and sequenced . As in Brucella cells, CP24 was located in the cytoplasm of recombinant Escherichia coli . The amino acid sequence predicted from the cloned gene revealed 48 and 46% identity with the ribosome releasing factor, a protein factor required for release of the 70S ribosome from the mRNA, of E . coli and Haemophilus influenzae Rd, respectively . Sera from naturally infected sheep and sheep experimentally infected with B . melitensis H38 showed antibody reactivity against recombinant CP24.

Infect Immun, 1996 Nov, 64(11), 4769 - 75
Effect of amplification of the Cap b locus on complement-mediated bacteriolysis and opsonization of type b Haemophilus influenzae; Noel GJ et al.; Amplification of the Cap b locus of Haemophilus influenzae occurs frequently in clinical isolates and has been proposed to be a mechanism by which this organism evades host defense . To determine if amplification of this locus affected complement fixation, in vitro studies to determine complement-mediated bacteriolysis and complement-mediated opsonization of an isogenic set of organisms containing 2, 3, and 4 copies of the Cap b locus were performed . Organisms containing 4 copies of the Cap b locus were significantly more resistant to antibody-dependent, classical complement pathway-directed bacteriolysis than were organisms containing 2 copies . Organisms containing 3 copies of this locus exhibited intermediate susceptibility to lysis . Complement-mediated opsonization of these organisms was assessed by determining the degree of binding of bacteria to murine or human macrophages or to nonphagocytic cells transfected with the genes for human Mac-1, the complement receptor type 3 . In all three assay systems, organisms containing 4 copies of the Cap b locus bound less well than did organisms containing 2 copies of this locus . Consistent with their decreased susceptibility to lysis and opsonization, organisms with 4 copies of the Cap b locus fixed less C3 than did organisms containing 2 copies . These data demonstrate that amplification of the Cap b locus is associated with decreased susceptibility to complement-mediated lysis and decreased complement-mediated opsonization and suggest that amplification is used by these pathogens to increase their resistance to complement-dependent host defense mechanisms {correction of mecanisms}.

Infect Immun, 1996 Nov, 64(11), 4673 - 9
Fine mapping of outer membrane protein P2 antigenic sites which vary during persistent infection by Haemophilus influenzae; Duim B et al.; Antigenic drift of the major outer membrane protein (MOMP) P2 of nonencapsulated Haemophilus influenzae as observed during persistent infections in patients with chronic bronchitis was mimicked in a rabbit model in which H . influenzae persisted in subcutaneous cages . The antigenic drift resulted from amino acid substitutions in potentially surface-exposed loops of MOMP P2 . Since in a rabbit model the appearance of antigenic variants was associated with the presence of strain-specific bactericidal antibodies (L . Vogel, B . Duim, F . Geluk, P . Eijk, H . Jansen, J . Dankert, and L . van Alphen, Infect . Immun . 64:980-986, 1996), we determined the epitope specificities of these bactericidal antibodies . The eight loops of MOMP P2 of H . influenzae d1 were separately expressed as fusion proteins with glutathione S-transferase . Sera of rabbits persistently infected with H . influenzae reacted with the loop 5 and loop 6 fusion proteins in immunoblotting and enzyme-linked immunosorbent assay . For fine mapping of the epitopes with pepscan analysis, overlapping synthetic peptides consisting of 12 amino acids were made . Rabbit sera contained antibodies reacting with peptides derived from loop 5 and peptides containing amino acids of the side of loop 6 . In addition, MOMP P2 variant-specific reactions with the amino acids located at the tip of loop 6 were detected . The rabbit sera showed variant-specific complement-dependent bactericidal activities, which were eliminated by affinity chromatography with fusion proteins of loop 6 but not of loop 5 . We conclude that, during persistence of H . influenzae in rabbits, variant-specific bactericidal antibodies are elicited to the variable tip of MOMP P2 loop 6.

Infect Immun, 1996 Nov, 64(11), 4618 - 29
Lack of expression of the global regulator OxyR in Haemophilus influenzae has a profound effect on growth phenotype; Maciver I et al.; A pBR322-based library of chromosomal DNA from the nontypeable Haemophilus influenzae TN106 was screened for the expression of transferrin-binding activity in Escherichia coli . A recombinant clone expressing transferrin-binding activity contained a 3.7-kb fragment of nontypeable H . influenzae DNA . Nucleotide sequence analysis of this insert revealed the presence of two complete open reading frames encoding proteins of approximately 26 and 34 kDa . Mini-Tn10kan transposon mutagenesis at different sites within the open reading frame encoding the 34-kDa protein resulted in the abolition of transferrin-binding activity in the recombinant E . coli clone . The deduced amino acid sequence of the 34-kDa protein had 70% identity with the OxyR protein of E . coli; this latter macromolecule is a member of the LysR family of transcriptional activators . When a mutated H . influenzae oxyR gene was introduced into the chromosome of the wild-type H . influenzae strain by allelic exchange, the resulting oxyR mutant still exhibited wild-type levels of transferrin-binding activity but was unable to grow on media containing the heme precursor protoporphyrin IX (PPIX) in place of heme . This mutant also exhibited reduced growth around disks impregnated with heme sources . Supplementation of the PPIX-based growth media with catalase or sodium pyruvate resulted in normal growth of the H . influenzae oxyR mutant . Provision of the wild-type H . influenzae oxyR gene in trans also permitted the growth of this mutant on a PPIX-based medium . Exogenously supplied catalase restored the growth of this mutant with heme sources to nearly wild-type levels . These results indicate that expression of a wild-type OxyR protein by H . influenzae is essential to allow this organism to protect itself against oxidative stresses in vitro.

Infect Immun, 1996 Nov, 64(11), 4586 - 92
Biological activity of serum antibodies to a nonacylated form of lipoprotein D of Haemophilus influenzae; Akkoyunlu M et al.; Protein D, a surface-exposed 42-kDa membrane lipoprotein, is well conserved among both type b and nontypeable Haemophilus influenzae strains, and it is considered a vaccine against H . influenzae infections . Here, we report the large-scale purification of a nonacylated form of protein D (PDm) from the periplasmic space of Escherichia coli overexpressing PDm . Screening of human sera for levels of antibodies to PDm demonstrated that the immunoglobulin G (IgG) antibody level is above background levels in infants less than 6 months of age . Following a drop to background values in the age group 6 months to 1 year, IgG antibody levels start to increase, together with IgA antibody levels, after 1 year of age . The first appearance of serum IgM antibodies is in 6-month- to 1-year-old infants whose IgG antibody levels have dropped to the postnatal background level . Affinity-purified antibodies from humans and from PDm-immunized rats detected epitopes of protein D which are normally exposed on the bacterial surface . Affinity-isolated human anti-PDm antibodies eluted in acidic buffer were not bactericidal against H . influenzae . Loss of bactericidal activity may occur in this buffer, as was demonstrated in pooled human sera with high bactericidal activity after incubation in the same buffer . Hyperimmunization of rats with PDm induced high levels of serum IgG and IgA antibodies against PDm and significant bactericidal activity against homologous and heterologous H . influenzae strains.

J Mol Evol, 1996 Nov, 43(5), 536 - 40
A modular domain of NifU, a nitrogen fixation cluster protein, is highly conserved in evolution; Hwang DM et al.; hnifU, a gene exhibiting similarity to nifU genes of nitrogen fixation gene clusters, was identified in the course of expressed sequence tag (EST) generation from a human fetal heart cDNA library . Northern blot of human tissues and polymerase chain reaction (PCR) using human genomic DNA verified that the hnifU gene represented a human gene rather than a microbial contaminant of the cDNA library . Conceptual translation of the hnifU cDNA yielded a protein product bearing 77% and 70% amino acid identity to NifU-like hypothetical proteins from Haemophilus influenzae and Saccharomyces cerevisiae, respectively, and 40-44% identity to the N-terminal regions of NifU proteins from several diazatrophs (i.e., nitrogen-fixing organisms) . Pairwise determination of amino acid identities between the NifU-like proteins of nondiazatrophs showed that these NifU-like proteins exhibited higher sequence identity to each other (63-77%) than to the diazatrophic NifU proteins (40-48%) . Further, the NifU-like proteins of non-nitrogen-fixing organisms were similar only to the N-terminal region of diazatrophic NifU proteins and therefore identified a novel modular domain in these NifU proteins . These findings support the hypothesis that NifU is indeed a modular protein . The high degree of sequence similarity between NifU-like proteins from species as divergent as humans and H . influenzae suggests that these proteins perform some basic cellular function and may be among the most highly conserved proteins.

J Mol Evol, 1996 Nov, 43(5), 453 - 9
Transkingdom transfer of the phosphoglucose isomerase gene; Katz LA; Previous analysis of the gene encoding phosphoglucose isomerase (Pgi) suggests that this gene may have been transferred between a eukaryote and a bacterium . However, excluding the alternative hypothesis of ancient gene duplication has proven difficult because of both insufficient sampling of taxa and an earlier misidentification of a bacterial Pgi sequence . This paper presents a phylogenetic analysis of published complete Pgi sequences together with analysis of new partial Pgi sequences from six species of bacteria . The data identify a group of bacterial Pgi sequences, including sequences from Escherichia coli and Haemophilus influenzae, which are more closely related to eukaryotic Pgi sequences than to other bacterial sequences . The topology of gene trees constructed using several different methods are all consistent with the hypothesis of lateral gene transfer and not ancient gene duplication . Furthermore, an estimate of a molecular clock for Pgi dates the divergence of the E . coli and H . influenzae sequences from the animal sequences to between 470 and 650 million years ago, well after other estimates of the divergence between eukaryotes and bacteria . This study provides the most convincing evidence to date of the transkingdom transfer of a nuclear gene.

Gene, 1996 Oct 31, 178(1-2), 89 - 96
The restriction-modification system of Pasteurella haemolytica is a member of a new family of type I enzymes; Highlander SK et al.; Genes encoding the type I restriction-modification (R-M) system of the bovine pathogen, Pasteurella haemolytica, have been identified immediately downstream of a locus that encodes a transcriptional activator of P . haemolytica leukotoxin expression . Type I enzymes are encoded by three genes called hsdM, hsdS and hsdR, and have fallen into three groups, called Ia, Ib and Ic . HsdS provides a sequence recognition function which in concert with HsdM forms an active methyltransferase (MTase) . Inclusion of the HsdR subunit in the complex creates an active restriction endonuclease (ENase) capable of cleaving unmethylated target DNA . The P . haemolytica hsdMSR genes were mapped using transposon Tn10d-Cam insertions, and bacteriophage restriction and modification assays in Escherichia coli . We determined the nucleotide sequences of hsdM, hsdS and hsdR, and observed that the deduced amino acid (aa) sequences were very similar to predicted R-M subunits in the respiratory pathogen, Haemophilus influenzae . Phylogenetic comparisons of all known Hsd aa sequences placed the P . haemolytica and H . influenzae proteins into a new group which we labeled the Type Id R-M family . Expression of the P . haemolytica R-M genes in E . coli was inefficient and is likely to be a consequence of the unusual codon usage in P . haemolytica genes.

Gene, 1996 Oct 31, 178(1-2), 75 - 81
Isolation and characterization of the Haemophilus influenzae tolQ, tolR, tolA and tolB genes; Sen K et al.; The tolQ, R, A and B genes have been isolated from the DNA of Haemophilus influenzae and sequenced . The deduced amino acid (aa) sequence of the H . influenzae TolQ, TolR, TolA and TolB show 67, 63, 41 and 62% identity with Escherichia coli TolQRAB proteins, respectively . These four proteins are involved in transport of colicins and phages across the cell envelope . The translational stop codon of TolB (the last gene in the cluster) is 23 bases upstream of the start codon of the P6 lipoprotein gene . Primer extension and Northern blot analysis revealed that the start of the P6 transcript is within the tolB gene . Nucleotide sequence (nt) analysis of the entire tolQRABP6 region shows a transcriptional terminator immediately downstream of the P6 gene . The tolQRABP6 gene cluster of H . influenzae may thus constitute an operon.

Stat Med, 1996 Oct 30, 15(20), 2235 - 52
A statistical model of transmission of Hib bacteria in a family; Auranen K et al.; The simultaneous estimation of family and community transmission rates as well as cure rates from panel data in a recurrent Hib (Haemophilus influenzae type b bacteria) infection is considered . An individual-based stationary Markov process model with constant hazards in two age groups is applied to describe recurrent asymptomatic Hib infection in a family with small children . The problem of estimation is solved in terms of the Bayesian posterior of the model parameters . The model is used to predict prevalence and incidence of Hib carriage in families as a function of the family size and age structure.

MMWR Morb Mortal Wkly Rep, 1996 Oct 25, 45(42), 901 - 6
Progress toward elimination of Haemophilus influenzae type b disease among infants and children--United States, 1987-1995; Isolation and characterization of the Haemophilus influenzae uvrA gene; Edward Mallinckrodt Department of Pediatrics, Washington University School of Medicine, St . Louis, MO 63110, USAThe uvrA gene Haemophilus influenzae (Hi) was cloned and sequenced . Analysis of the deduced amino acid sequence revealed 81% identity and 90% similarity with the Escherichia coli UvrA protein . Consistent with a role of Hi uvrA in DNA repair, a Hi uvrA mutant exhibited increased sensitivity of UV irradiation . Furthermore, Hi uvrA was able to complement a mutation in the E . coli uvrA locus.

Gene, 1996 Oct 17, 176(1-2), 125 - 9
Cloning and sequencing of a Haemophilus ducreyi fur homolog; Carson SD et al.; Iron regulation in a growing number of bacterial species is being attributed to the presence of a fur (ferric uptake regulation) regulatory system . In the presence of iron, Fur acts as a classical negative regulator, binding conserved sequences within the promoter of iron-repressible genes and blocking transcription . Western blot analysis utilizing Escherichia coli Fur antisera detected a band of approximately 17 kDa in soluble extracts of Haemophilus ducreyi . Additionally, Southern blot hybridization of the H . ducreyi chromosome with a meningococcal fur probe indicated that H . ducreyi might contain a fur homolog . This putative fur homolog was cloned into the E . coli vector pACYC184 . This clone was capable of repressing expression of a normally Furregulated lacZ fusion in the fur-background of E . coli strain H1780 . The deduced amino acid sequence shows H . ducreyi fur to be 54% identical and 73% similar to E . coli fur, containing putative DNA-binding and metal-binding domains . These data demonstrate that H . ducreyi has a functional fur system.

Proc Natl Acad Sci U S A, 1996 Oct 15, 93(21), 11913 - 8
Haemophilus influenzae pili are composite structures assembled via the HifB chaperone; St Geme JW et al.; Haemophilus influenzae is a Gram-negative bacterium that represents a common cause of human disease . Disease due to this organism begins with colonization of the upper respiratory mucosa, a process facilitated by adhesive fibers called pili . In the present study, we investigated the structure and assembly of H . influenzae pili . Examination of pili by electron microscopy using quick-freeze, deep-etch and immunogold techniques revealed the presence of two distinct subassemblies, including a flexible two-stranded helical rod comprised of HifA and a short, thin, distal tip structure containing HifD . Genetic and biochemical studies demonstrated that the biogenesis of H . influenzae pili is dependent on a periplasmic chaperone called HifB, which belongs to the PapD family of immunoglobulin-like chaperones . HifB bound directly to HifA and HifD, forming HifB-HifA and HifB-HifD complexes, which were purified from periplasmic extracts by ion-exchange chromatography . Continued investigation of the biogenesis of H . influenzae pili should provide general insights into organelle development and may suggest novel strategies for disease prevention.

Gene, 1996 Oct 10, 175(1-2), 137 - 41
Cloning and expression of genes encoding lipid A biosynthesis from Haemophilus influenzae type b; Servos S et al.; Genes similar to Escherichia coli lpx genes (encoding enzymes required for the biosynthesis of lipid A) have been cloned from Haemophilus influenzae type b using a hybridisation-based strategy . The derived amino acid sequences are highly homologous to their E . coli counterparts . The genes appear in the same order in both E . coli and H . influenzae, but the intergenic regions differ . H . influenzae lpxA and lpxB have been expressed in E . coli minicells and they encode proteins of the predicted sizes . Both H . influenzae lpxA and lpxB are able to complement temperature-sensitive mutants in the equivalent genes in E . coli . This provides evidence that the genetic manipulation of lpx genes to generate altered lipid A molecules may be possible.

JAMA, 1996 Oct 9, 276(14), 1181 - 5
The 1996 Albert Lasker Medical Research Awards . Prevention of systemic infections, especially meningitis, caused by Haemophilus influenzae type b . Impact on public health and implications for other polysaccharide-based vaccines; Robbins JB et al.; The development of Haemophilus influenzae type b (Hib) conjugate vaccines has led to the virtual elimination of systemic infections caused by that pathogen, has provided insights into the pathogenesis of and immunity to other capsulated bacteria, and has contributed to the development of new vaccines . Meningitis, a common and serious infection of children, and other infections caused by Hib have been virtually eliminated in countries that have achieved widespread vaccination with Hib conjugates, including the United States, Canada, the United Kingdom, Iceland, Scandinavia, France, and Germany . Hib conjugates have also been shown to be highly effective in developing countries . The principles derived from the use of these vaccines, along with studies of other capsulated pathogens, should allow the rapid inclusion of new polysaccharide-based conjugates into routine vaccination schedules of infants, and should help to realize further reductions in serious systemic infectious diseases.

Enferm Infecc Microbiol Clin, 1996 Oct, 14(8), 483 - 5
{Urinary tract infection caused by Haemophilus spp . in pediatrics: a rarely studied disease}; Galan F et al.; BACKGROUND: Haemophilus influenzae and Haemophilus parainfluenzae have been, in very seldom times, described as a cause of urinary tract infection (UTI), mainly on adults . Their real incidence has been scarcely studied . In view of this situation, we have investigated the cases occurred during the year of 1994 in our hospital, in Cadiz (Spain) . METHODS: A total of 16,446 urine cultures from in-patients and ambulatory patients were processed, including the chocolate agar medium in the cultures . RESULTS: Urinary infection by Haemophilus supposed an incidence of 0.27% with respect to all positive urine cultures, and a 0.88% of the paediatric ones . H . influenzae was isolated in 7 occasions, and H . parainfluenzae in 3 of them . The 80% of the cases of UTI corresponded to paediatric patients . CONCLUSIONS: It would be convenient the inclusion of chocolate agar medium in the urine cultures for determined patients, chiefly on children.

An Esp Pediatr, 1996 Oct, 45(4), 380 - 5
{Pneumonia due to Haemophilus influenzae type b in children: presentation of 24 cases}; Otero Reigada MC et al.; OBJECTIVE: Due to the higher incidence in invasive infections, the changes observed in the sensitivity of this microorganism during the last decade and the seriousness of pulmonary infections in children, the objective of this paper is to review the clinical and epidemiological characteristics of Haemophilus influenzae type b (Hib) pneumonia, as well as to give an update of treatment . PATIENTS AND METHODS: Twenty-four children diagnosed between 1973 and 1992 are reviewed using the following criteria: clinical manifestations of pneumonia, radiology and isolation of the microorganism . RESULTS: The annual average was 7.8 cases/100,000 children under 5 years of age . More females were affected (58.3% vs 41.6%) . Of the cases studied, 58.3% of the children were under one year and 83.3% under 2 years of age . Fever was the most common clinical manifestation (95.8%) . Blood cultures were positive in 21 cases (87.5%); pleural effusion culture contributed to the diagnosis in two other cases (8.3%) and deep bronchial aspirate in one other case . Of the Haemophilus influenzae type b strains 62.5% were resistant to ampicillin and 100% were sensitive to second and third generation cephalosporins . Three infants with chronic illness died (12.5%) . CONCLUSIONS: Hib pneumonia is an infection usually affecting children younger than 2 years of age . Its clinical manifestations are indistinguishable from those of other microorganisms . Blood culture is the most useful technique to confirm the diagnosis . The increase in resistance to classical treatment (ampicillin) obliges us to consider amoxicillin-clavulanic as the selected empirical treatment . Second and third generation cephalosporins may be considered a good alternative . There is a high mortality in children with chronic illness.

Vaccine, 1996 Oct, 14(15), 1471 - 8
Modulation of antigen-specific T and B cell responses influence bacterial clearance of non-typeable Haemophilus influenzae from the lung in a rat model; Kyd JM et al.; This study investigates antigen-specific B and T cell responses following mucosal immunization with the major outer membrane protein, P2, from non-typeable Haemophilus influenzae (NTHi) and the role of these responses in bacterial clearance following pulmonary challenge . Modification of the immunization preparation by the inclusion of sodium dodecyl sulphate (SDS) with the adjuvant, incomplete Freund's, differentially affects the T and B cell responses to the P2 antigen . Rats received an intra-Peyer's patch-immunization with P2 with or without the inclusion of 1% (w/v) SDS, were boosted via an intratracheal administration of P2 alone on day 14, and challenged with live NTHi in the lungs on day 21 . There were significant differences in the rate of bacterial clearance between the different P2-immunized groups and the non-immune group . The inclusion of SDS with P2 resulted in enhanced bacterial clearance . This clearance corresponded to an enhancement of P2-specific lymphocyte proliferation by CD4+ T helper cells but a decrease (reduced approximately 75%) in anti-P2 IgG and IgA in both serum and bronchoalveolar lavage washings . P2-specific IgM levels were not altered . IgG subclass analysis indicated that the inclusion of SDS had caused a significant reduction in IgG2a and an increase in IgG1 . The data indicates that the magnitude of antibody levels to P2 may not be as important as T cell responses in enhancing clearance of NTHi in the lung, in vivo, and that immunization targeting enhancement of antigen-specific T cells may be important to inducing effective immunity to NTHi.

Vaccine, 1996 Oct, 14(15), 1399 - 406
Study of booster doses of two Haemophilus influenzae type b conjugate vaccines including their interchangeability; Scheifele D et al.; A prospective, two center study of 319 children was undertaken to assess responses to booster immunization in healthy 18-month-olds who completed primary immunization 12 months earlier with Haemophilus influenzae type b (Hib) conjugate vaccine (either HbOC or PRP-T) . Interchangeability of these products as boosters was also assessed, using combination products containing diphtheria, pertussis and tetanus components . The study was randomized and evaluator blinded . Sera obtained prior to booster vaccination and 4 weeks later were tested for anti-PRP using an IgG-specific enzyme immunoassay . Pre-immunization anti-PRP levels were low: unmeasurable levels (< 0.06 microgram ml-1) were present in 47% of those primed with HbOC and 35% of those primed with PRP-T (P < 0.05) . Most children responded strongly to booster vaccination . Interchanging the products had no detrimental effect . Children who lacked measurable antibody prior to re-vaccination had significantly weaker responses 4 weeks afterward . Earlier Hib booster vaccination should be considered in Canada to avoid low anti-PRP trough levels . Extended follow-up of antibody levels and vaccine protective efficacy is also advisable given the observed heterogeneity of responses to booster vaccination.

Turk J Pediatr, 1996 Oct-Dec, 38(4), 407 - 12
Microorganisms involved in acute bacterial meningitis in children and the role of Haemophilus influenzae; Kanra G et al.; Acute bacterial meningitis (ABM) is an important cause of mortality and neurological damage in children . Documentation of the etiological agents is very important both for the treatment of patients and for prophylactic approaches . H.influenzae, N.meningitidis and S.pneumoniae are the three major pathogens involved in ABM . In Turkey for many years H.influenzae has not been isolated in cerebrospinal fluid (CSF) specimens . In order to show the bacteria involved in ABM in our hospital and to see the role of H.influenzae, we investigated the CSF of 59 patients with bacterial meningitis using Gram and Wayson stains, culture and latex agglutination techniques . The agents were determined in 38 (64.4%) specimens by using culture positivity in 30 (50.8%), and latex or stain positivity in eight (13.6%) specimens . The microorganisms causing ABM included S.pneumoniae (25.6%), gram-negative enteric bacillI (17.9%), N.meningitis (12.8%), alpha hemolytic streptococci (10.3%), H.influenzae (10.3%), nonfermentative gram-negative bacillI (5.1%), candida spp . (5.1%), group B streptococci (2.6%), coagulase negative staphylococci (2.6%), S.aureus (2.6%) and pseudomonas spp . (5.1%) . In this study it has been shown that H.influenzae can cause ABM in Turkish children . Multicentric studies from different parts of Turkey will be helpful in showing the real incidence in our country.

Jpn J Antibiot, 1996 Oct, 49(10), 939 - 46
{Clinical studies on azithromycin in pediatrics}; Okamoto T et al.; Fine granules or capsules of azithromycin (AZM) were given to 32 pediatric patients for the treatment of the following diseases: pharyngitis in three cases; tonsillitis in one; bronchitis in six; pneumonia in six; mycoplasmal pneumonia in 14; pertussis and enteritis in one, each . Effectiveness of AZM was evaluated in 30 cases and the drug was rated "excellent" in 18 patients, "good" in 11 and "fair" in one, resulting in a total efficacy rate of 96.7% . Three strains of bacteria were isolated from 3 patients as the causative organisms including: Streptococcus pneumoniae, Haemophilus influenzae and Haemophilus parainfluenzae, from three different patients, respectively . One patient complained of mild diarrhea, another patient mild urticaria . Abnormal laboratory test results were reported as follows: one patient showed a slight decrease in leukocyte count, three patients showed slight increases in eosinophils, and one patient had slight elevations in GOT and GPT . The above results suggest that AZM is a useful antibiotic drug in the treatment of pediatric patients with various bacterial infections.

Jpn J Antibiot, 1996 Oct, 49(10), 926 - 31
{Therapeutic efficacy of azithromycin in pediatrics}; Haruta T et al.; Azithromycin (AZM), a newly developed azalide antibiotic, was administered at a standard dose of 10 mg/kg once daily for 3 days to pediatric patients with bacterial infections and the therapeutic efficacy of AZM was investigated . 1 . A total of 12 patients with the following diseases was evaluated: pharyngitis in two, tonsillitis in four, bronchitis in one, Mycoplasma pneumonia in one, scarlet fever in two and enteritis in two . The drug was rated "excellent" in eight cases and "good" in four . 2 . Eleven strains were isolated from patients: five strains of Streptococcus pyogenes, four strains of Haemophilus influenzae, and two strains of Haemophilus parainfluenzae . Isolated bacteria were eradicated in eight strains and persisting in one, resulting in 88.9% in eradication rate . No follow-up examinations in post-treatment were performed in two cases . 3 . No adverse reaction was reported, while one case of eosinophilia was noted as an abnormal laboratory test value . 4 . As far as compliance is concerned, patients claimed that the formulation of the drug is "easy to take" or "ordinary" . With the results presented as above, we have concluded that AZM is a useful antibiotic in pediatric patients with bacterial infections.

Diagn Microbiol Infect Dis, 1996 Oct, 26(2), 95 - 8
Nontypeable Haemophilus influenzae susceptibility: effect of inoculum size and beta-lactamase production; Gould JM et al.; The activities of cefixime, cefpodoxime, cefprozil, cefuroxime, loracarbef, and amoxicillin/clavulanate against 72 clinical isolates of nontypeable Haemophilus influenzae were determined by using an agar dilution method . The effects of beta-lactamase production and bacterial inoculum size were investigated . All antimicrobials exhibited a significant inoculum effect, demonstrating the importance of accurately determining inoculum size in the performance of antimicrobial susceptibility testing of H . influenzae.

Arch Dis Child, 1996 Oct, 75(4), 298 - 303
Combined diphtheria, tetanus, pertussis, and Haemophilus influenzae type b vaccines for primary immunisation; Bell F et al.; A total of 146 infants were immunised at ages 2, 3, and 4 months with a combined diphtheria, tetanus, pertussis (DTP)--Haemophilus influenzae type b (Hib) tetanus toxoid conjugate (PRP-T) vaccine (Pasteur Merieux) to assess the antibody response and adverse events associated with immunisation . Adverse events, including fever, were recorded by parents in a diary for three days following each injection . Blood was taken before the first immunisation and four weeks after the third immunisation to assess antibody response . Data were compared with those from historical controls who had received DTP and PRP-T vaccines by separate injection . The combined vaccine was well tolerated . Rates of local and general reactions were similar to those reported for infants immunised by separate injection . All infants achieved protective antibody titres (> 0.01 IU/ml) for diphtheria and tetanus; 98% acquired Hib (PRP) antibody > 0.15 microgram/ml and 82.5% > 1.0 microgram/ml . Pertussis antibody titres (pertussis toxin, filamentous haemagglutinin, total agglutinins, and agglutinins 2 and 3) showed appreciable rise following immunisation . DTP and PRP-T vaccines provide similar antibody responses and adverse effects whether mixed in the same syringe or administered by separate injection . The vaccines could be combined for use in the United Kingdom primary immunisation schedule.

APMIS, 1996 Oct, 104(10), 709 - 17
Local and systemic antibody levels against protein D of Haemophilus influenzae following immunization and infection in rats; Akkoyunlu M et al.; Outer membrane proteins of Haemophilus influenzae are considered as possible vaccine candidates against non-typable H . influenzae (NTHi), a major cause of respiratory infections . Here, we study local and systemic antibody responses to protein D, a well-conserved 42-kDa membrane protein, following local and systemic immunization, and experimental acute otitis media (AOM) with NTHi and H . influenzae type b (Hib) in rats . Animals that were challenged and rechallenged in the middle ear with Hib strain Minn A or NTHi strain 1161 developed IgG and IgA antibodies in serum but not in middle ear lavage (MEL) material or saliva . In contrast, following per oral immunization with NTHi strain 772 and Escherichia coli JM83 (pHIC348) (containing protein D gene) and, to a lesser degree after intranasal inoculation of NTHi strain 772, high saliva IgA antibodies to protein D developed, but there was no rise in antibodies to protein D in the MEL material or the sera of these animals . These results show that protein D can elicit different systemic and local antibody responses depending on the site of delivery and the form of administration . Furthermore, experimental AOM with NTHi and Hib induces systemic IgG and IgA antibodies to protein D but fails to induce a mucosal immune response.

Thorax, 1996 Oct, 51(10), 1010 - 6
Community acquired pneumonia: aetiology and usefulness of severity criteria on admission; Neill AM et al.; BACKGROUND: Community acquired pneumonia remains an important cause of hospital admission and carries an appreciable mortality . Criteria for the assessment of severity during admission have been developed by the British Thoracic Society (BTS) . A study was performed to determine the sensitivity and specificity of a severity rule based on a modification of the BTS prognostic rules applied on admission, to compare severity as assessed by medical staff with the modified rule, and to determine the microbiological cause of community acquired pneumonia in Christchurch . METHODS: A 12 month study of all adults admitted to Christchurch Hospital with community acquired pneumonia was undertaken . Three hundred and sixteen consecutive patients with suspected community acquired pneumonia were screened for inclusion . Variables obtained from the history, examination, investigations, and initial treatment were examined for association with mortality . RESULTS: Two hundred and fifty five patients met the inclusion criteria . Their mean age was 58 years (range 18-97) . A microbiological diagnosis was made in 181 cases (71%), Streptococcus pneumonia (39%), Mycoplasma pneumoniae (16%), Legionella species (11%), and Haemophilus influenzae (11%) being the most commonly identified organisms . Patients had a 36-fold increased risk of death if any two of the following were present on admission: respiratory rate > or = 30/min, diastolic BP < or = 60 mm Hg, urea > 7 mmol/l, or confusion . The severity rule identified 19 of the 20 patients who died and six of eight patients admitted to the intensive care unit as having life threatening community acquired pneumonia . The sensitivity of the modified rule for predicting death was 0.95 and the specificity 0.71 . In 47 cases (21%) the clinical team appeared to underestimate the severity of the illness . CONCLUSIONS: The organisms responsible for community acquired pneumonia in Christchurch are similar to those reported from other centres except for Legionella species which were more common than in most studies . The modification of the BTS prognostic rules applied as a severity indicator at admission performed well and could be incorporated into management guidelines.

J Antibiot (Tokyo), 1996 Oct, 49(10), 1036 - 43
Preparation and in vitro antibacterial activity of 9-O-glycosyloxime derivatives of erythromycin A, a new class of macrolide antibiotics; Grandjean C et al.; 9-O-Glycosyloxime derivatives of erythromycin A have been synthesized and their in vitro antibacterial activity compared with that of erythromycin A (1) . This new class of macrolide antibiotics showed reduced antibacterial spectrum . However, some derivatives were as or more active than erythromycin A (1) against strains, responsible for respiratory track infections, such as Haemophilus influenzae, Moraxella catarrhalis or Streptococcus pneumoniae.

Eur J Clin Microbiol Infect Dis, 1996 Oct, 15(10), 820 - 3
Meningitis due to enterotoxigenic Bacteroides fragilis; Aucher P et al.; An enterotoxigenic strain of Bacteroides fragilis was the sole organism isolated from the cerebrospinal fluid of a two-and-a-half-month neonate who had a medullary-colonic fistula as part of a complex congenital malformation, but no brain abscess . A rapid latex particle agglutination test for detection of bacterial antigen was positive for Haemophilus influenzae type b, suggesting that Bacteroides fragilis and Haemophilus influenzae type b might share some capsular antigens . In order to determine the role of the enterotoxin with respect to virulence of the strain, antibodies to a 20 kDa protein were sought in the patient's serum, but Western blot of the culture supernatant revealed only antibodies to a 45 kDa bacterial protein . The patient was successfully treated with metronidazole and imipenem.

J Antimicrob Chemother, 1996 Oct, 38(4), 713 - 8
In-vitro activity of teicoplanin and ceftriaxone in combination against pathogens involved in paediatric bone and joint infections; Lhopital S et al.; We investigated the combination of teicoplanin and ceftriaxone for its bactericidal activity against Staphylococcus aureus, Haemophilus influenzae type b and Streptococcus pneumoniae isolated from bone and joint infections in children . An increase in bactericidal activity was observed against isolates of S . aureus and H . influenzae when the antibiotics were tested at fractional MICs whereas indifference was observed at their MICs . Similar results were obtained at fractional MICs against S.pneumoniae, but the bactericidal activity fell by more than 1 x log 10 cfu/mL when the antibiotics were tested at or above their MICs.

J Antimicrob Chemother, 1996 Oct, 38(4), 679 - 89
Analysis of oral antibiotic treatment that failed to prevent the development of Haemophilus influenzae meningitis: consequences on mortality; Dabernat H et al.; One hundred and four children suffering from Haemophilus influenzae meningitis were studied in a multi-centre study in France between October 1991 and June 1993 . The clinical and bacteriological findings were similar to those usually found in this condition . Fifty three per cent of the H . influenzae strains cultured produced beta-lactamase and 98% were of type b . Forty-six children who had been prescribed oral antibiotics before hospitalisation were analysed in this study . This analysis took into account the nature of the antibiotic and whether it was prescribed as the treatment of first or second intention . Treatment failures, defined according to pre-established criteria, were found to be independent of whether or not the incriminated strain was beta-lactamase producing and whether or not it was sensitive in vitro to the antibiotic prescribed . None of the 46 children who had received pre-hospital oral antibiotics died while three of the 58 who had received pre-hospital oral antibiotics died . However, it is difficult to establish in our study a clear relationship between the reduction of mortality and previous oral antibiotic treatment in bacteriologically proven cases of H . influenzae meningitis.

J Antimicrob Chemother, 1996 Oct, 38(4), 627 - 39
A multinational European survey on the in-vitro activity of rufloxacin and other comparative antibiotics on respiratory and urinary bacterial pathogens; Schito GC et al.; The antibacterial activity of rufloxacin was confirmed against a large number of respiratory and urinary tract pathogens collected in five European countries . In terms of both MIC90 values and percentages of susceptible isolates found, this new quinolone showed useful in-vitro activity against Mycoplasma catarrhalis, Haemophilus influenzae and Klebsiella pneumoniae, with a large proportion of Staphylococcus aureus also covered, while, as expected Streptococcus pneumoniae and Streptococcus pyogenes were not included in its antibacterial spectrum . Rufloxacin was comparable with the other antibiotics against these pathogens with the exception of streptococci . Against these microorganisms, beta-lactams were the most active agents . Against the urinary pathogens the in-vitro activity of rufloxacin is similar to that of norfloxacin although this latter drug is more active in terms of MIC90 values and in percentages of strains inhibited . In many cases more isolates were susceptible to the remaining comparator agents than to rufloxacin . However, there were no significant differences between the numbers of microorganisms inhibited by the various drugs and rufloxacin . The findings of this survey do not seem to modify the general picture which emerged in previous studies and further confirms its useful spectrum in different geographic settings.

J Antimicrob Chemother, 1996 Oct, 38(4), 615 - 25
Antimicrobial resistance and epidemiological study of Haemophilus influenzae strains isolated in Portugal . The Multicentre Study Group; Bajanca-Lavado MP et al.; In the course of a multicentric surveillance study, nine laboratories sent 375 isolates of Haemophilus influenzae to the Sector de Resistencia aos Antibioticos (SRA) from the National Institute of Health in Lisbon, between 1 January and 31 December 1992 . The majority of the H . influenzae isolates were from the respiratory tract (84.8%); only 5.1% were of invasive origin . Overall resistance for ampicillin was 11.7%, tetracycline 3.7%, and chloramphenicol 2.4% . All isolates tested were fully susceptible to cefotaxime, ceftriaxone, ciprofloxacin and rifampicin . Multiresistance was rare, occurring only in 2.4% of the isolates, although 50% of the ampicillin resistant strains had at least one additional resistance marker . Forty two isolates (11.2%) produced a TEM-1 type beta-lactamase, as shown by isoelectric focusing . beta-lactamase production was not detected in two of the ampicillin resistant strains . Fifteen of the 42 beta-lactamase producing strains (35.7%) contained detectable DNA plasmid: nine harboured large plasmids with an apparent molecular mass of 45 or 54 kb depending on their resistance phenotype and six harboured a small plasmid of 5 kb . In order to study transfer of resistance in both ampicillin and multiresistant strains conjugation experiments were performed for 14 isolates, seven of which harboured a large plasmid and seven had no detectable plasmid DNA . All 14 transferred their resistance phenotype but only a single large plasmid could be demonstrated in ten transconjugants . Restriction endonuclease analysis of plasmids from six representative transconjugants, isolated in different hospitals, revealed that there was no dissemination of a single R plasmid, which suggests an independent process of acquisition of resistance genes.

J Trop Pediatr, 1996 Oct, 42(5), 256 - 61
Pediatric meningitis in the Western Cape Province of South Africa; Donald PR et al.; During the 9 years 1985-1993 a prospective survey of all cases of meningitis in children < 13 years of age presenting to our hospital in the Western Cape Province of South Africa was carried out . Two-thousand-nine-hundred-and-twenty cases of meningitis were identified . The commonest form of bacterial meningitis was tuberculous meningitis (TBM) diagnosed in 282 children (mean age 2.94 years) . N . meningitidis identified in 220 children (mean age 2.87 years), Haemophilus influenzae in 156 children (mean age 1.15 years) and S . pneumoniae in 106 children (mean age 2.14) were the next commonest causes of bacterial meningitis diagnosed . One-hundred-and-eighteen cases of bacterial meningitis were confirmed in infants < 1 month of age and the commonest bacteria identified were group B beta-haemolytic Streptococcus in 27, E . coli in 21, Klebsiella species in 11, and Candida species in 15 neonates . The emergence of TBM as the predominant cause of bacterial meningitis in childhood at our hospital is probably a reflection of the worsening tuberculosis situation in the Western Cape Province of South Africa.

Infect Immun, 1996 Oct, 64(10), 4339 - 44
Prolonged and preferential production of polymeric immunoglobulin A in response to Streptococcus pneumoniae capsular polysaccharides; Johnson S et al.; Streptococcus pneumoniae is an invasive mucosal pathogen for which host defense is dependent on capsular polysaccharide-specific antibody . Capsule-specific immunoglobulin G (IgG), IgM, and IgA are produced following pneumococcal vaccination and infection . Serum IgA has two molecular forms, polymeric and monomeric . These forms may modulate the avidity of antigen binding and evolve over time as the immune response matures . Therefore, we sequentially characterized the molecular forms of serum IgA to three serotypes of pneumococcal capsular polysaccharides (types 8, 12F, and 14) after pneumococcal vaccination and after natural infection with type 14 S . pneumoniae . Although typically the form of IgA in antigen-specific systemic responses to protein antigens is predominantly polymeric in sera of patients shortly after exposure and shifts to the monomeric form in sera obtained several weeks later, the form of IgA in response to each pneumococcal capsular polysaccharide remained predominantly polymeric 1 month after natural infection and up to I year following vaccination . In contrast, IgA to pneumococcal cell wall polysaccharide was both polymeric and monomeric . Moreover, the form of IgA in response to polyribosyl-ribitol-phosphate (PRP), the capsular polysaccharide of Haemophilus influenzae type b, was predominantly monomeric in the sera of 8 of 10 subjects tested 1 to 3 months after vaccination with either PRP alone or the diphtheria toxoid conjugate of PRP . We conclude that systemic responses to pneumococcal capsular polysaccharides are distinct in the production of predominantly polymeric IgA over time . The persistence of polymeric IgA may facilitate binding and clearance of pneumococci from the systemic circulation or reflect limited maturation of the immune response to pneumococcal capsular polysaccharides.

Infect Immun, 1996 Oct, 64(10), 4047 - 53
Synthesis, characterization, and immunologic properties of detoxified lipooligosaccharide from nontypeable Haemophilus influenzae conjugated to proteins; Gu XX et al.; Nontypeable Haemophilus influenzae (NTHi) is an important cause of otitis media in children and of pneumonitis in adults with depressed resistance . Lipooligosaccharide (LOS) is a major surface antigen of NTHi and elicits bactericidal and opsonic antibodies . We prepared detoxified LOS (dLOS) protein conjugates from NTHi for use as experimental vaccines . LOS from NTHi 9274 was treated with anhydrous hydrazine and had its toxicity reduced to clinically acceptable levels . dLOS was bound to tetanus toxoid (TT) or high- molecular-weight proteins (HMPs) from NTHi through a linker of adipic acid dihydrazide to form dLOS-TT or dLOS-HMP . The molar ratio of the dLOS to protein carriers ranged from 26:1 to 50:1 . The antigenicity of the conjugates was similar to that of the LOS alone as determined by double immunodiffusion . Subcutaneous or intramuscular injection of the conjugates elicited a 28- to 486-fold rise in the level of immunoglobulin G antibodies in mice to the homologous LOS after two or three injections and a 169- to 243-fold rise in the level of immunoglobulin G antibodies in rabbits after two injections . The immunogenicity of the conjugates in mice and rabbits was enhanced by formulation with monophosphoryl lipid A plus trehalose dimycolate . In rabbits, conjugate-induced LOS antibodies induced complement-mediated bactericidal activity against the homologous strain 9274 and prototype strain 3189 . These results indicate that a detoxified LOS-protein conjugate is a candidate vaccine for otitis media and pneumonitis caused by NTHi.

J Vet Med Sci, 1996 Oct, 58(10), 1007 - 9
Characterization and pathogenicity of Haemophilus paragallinarum serotype C in local chickens of Taiwan; Lin JA et al.; Two strains (Strains 39 & B) of Haemophilus paragallinarum were isolated and identified as serotype C by dot-blotting analysis in Taiwan in 1994 . To identify their virulence and pathogenicity in Taiwanese local chickens, local chickens were inoculated via nostril with the isolates, and examined for 14 days . Clinical signs of swollen face, and nasal discharge were first seen at 24 hr after inoculation, and the infectivity of Strains 39 and B at 14 days after inoculation among the inoculated chickens was 50.0-71.4% and 55.6-77.8%, respectively . From the results, we concluded that the two bacterial isolates were pathogenic to local chickens with a maximum morbidity of 77.8%.

Clin Infect Dis, 1996 Oct, 23(4), 723 - 8
Severe community-acquired pneumonia in the elderly: epidemiology and prognosis . Study Group for Severe Community-Acquired Pneumonia; Rello J et al.; Ninety-five patients with severe community-acquired pneumonia (SCAP) who were > or = 65 years of age were studied prospectively . A definite pathogen was identified in 37 cases (38.9%) and was most commonly Streptococcus pneumoniae, Haemophilus influenzae, or another gram-negative bacillus . The overall death rate was 40% . Eighty-three patients required mechanical ventilation and 40 needed vasoactive drugs . Multivariate analysis showed that the risk of death was higher in cases involving rapid radiological spread (relative risk {RR} = 6.99; 95% confidence interval (95% CI) = 1.54-31.70), shock (RR = 6.70; 95% CI = 2.13-21.02), previous steroid treatment or immunosuppression (RR = 5.50; 95% CI = 0.77-39.10), acute renal failure (RR = 3.88; 95% CI = 1.30-11.59), or an APACHE II score of > 22 on admission (RR = 2.25; 95% CI = 0.73-6.95) . We conclude that SCAP in elderly patients is associated with high mortality, but it is inappropriate to withhold intensive care on account of age . The presence of complications and the severity of illness at initial presentation were the major variables affecting outcome . Except for immunosuppression, comorbidities did not seem to influence outcome . Finally, our data reinforce the current American Thoracic Society guidelines concerning therapy for patients with SCAP.

J Clin Microbiol, 1996 Oct, 34(10), 2613 - 5
Ribosomal DNA typing of Haemophilus ducreyi strains: proposal for a novel typing scheme; Pillay A et al.; Local strains of Haemophilus ducreyi were characterized by ribosomal DNA fingerprinting in order to develop a typing scheme for this organism . Chromosomal DNAs from 30 strains were extracted, and restriction fragments were separated by agarose gel electrophoresis after cleavage with BglII, HindIII, HindII, and BstEII . Of the 30 local strains studied, nine BglII, eight HindIII, six HindII, and five BstEII ribotype patterns were observed . In order to develop a typing scheme, we correlated the ribotype patterns obtained with each of the four enzymes for each strain . By this scheme, 13 ribotypes designated b to n were defined among the local strains . This study demonstrates that there is a diversity of ribotypes among H . ducreyi strains in our area, which emphasizes that there are many strains responsible for chancroid . This novel typing scheme provides a useful tool for further epidemiological studies of chancroid.

J Clin Microbiol, 1996 Oct, 34(10), 2483 - 92
GlpQ: an antigen for serological discrimination between relapsing fever and Lyme borreliosis; Schwan TG et al.; Tick-borne relapsing fever is caused by numerous Borrelia species maintained in nature by Ornithodoros tick-mammal cycles . Serological confirmation is based on either an immunofluorescence assay or an enzyme-linked immunosorbent assay using whole cells or sonicated Borrelia hermsii as the antigen . However, antigenic variability of this bacterium's outer surface proteins and antigens shared with the Lyme disease spirochete (B . burgdorferi), may cause both false-negative and false-positive results when testing sera of patients suspected to have either relapsing fever or Lyme disease . To develop a specific serological test for relapsing fever, we created a genomic DNA library of B . hermsii, screened transformed Escherichia coli cells for immunoreactivity with high-titered (> or = 1:2,048) human anti-B . hermsii antiserum, and selected an immunoreactive clone (pSPR75) expressing a 39-kDa protein . DNA sequencing, subcloning, and serum adsorption experiments identified the immunoreactive protein as a homolog of GlpQ, a glycerophosphodiester phosphodiesterase identified previously in E . coli, Haemophilus influenzae, and Bacillus subtilis . Serum samples from humans and mice infected with B . hermsii or other species of relapsing fever spirochetes contained antibodies recognizing GlpQ, whereas serum samples from Lyme disease and syphilis patients were nonreactive . Serologic tests based on this antigen will identify people exposed previously to relapsing fever spirochetes and help clarify the distribution of relapsing fever and Lyme disease in situations in which the occurrence of their causative agents is uncertain.

J Clin Microbiol, 1996 Oct, 34(10), 2426 - 31
Haemophilus influenzae adherent to contact lenses associated with production of acute ocular inflammation; Sankaridurg PR et al.; Ten episodes of adverse responses to contact lens wear, including contact lens-induced acute red eye (CLARE), in which Haemophilus influenzae was isolated from contact lenses and/or from one of the external ocular sites at the time of the event, are described . All episodes occurred in patients wearing disposable hydrogel lenses on a 6-night extended-wear schedule . Two of the patients had recurrent episodes . H . influenzae was usually isolated in large numbers, and other bacteria or fungi colonizing the contact lens or the external ocular surface were usually present in low numbers . Those patients who were colonized with H . influenzae were more than 100 times as likely to have had a CLARE or infiltrative response than those subjects who were not colonized with this bacterium . H . influenzae colonization of the contact lens and eye may be subsequent to colonization of the nasopharynx because four of the seven patients presented with fever at the time of the event, with concurrent upper respiratory tract infection . Contact lens wearers should be made aware of the potential risk of CLARE associated with the wearing of contact lenses for extended periods during and subsequent to upper respiratory tract infection.

Am J Respir Crit Care Med, 1996 Oct, 154(4 Pt 2), S192 - 6
Molecular determinants of the interaction between Haemophilus influenzae and human cells; Geme JW 3rd; Haemophilus influenzae is a human-specific pathogen that must colonize the human upper respiratory tract to avoid extinction . On occasion, organisms penetrate the epithelial barrier and cause bacteremic disease or spread within the respiratory tract to produce localized disease . Attachment to host epithelium is fundamental to the process of colonization and to the pathogenesis of disease . Accordingly, H . influenzae has evolved to express a number of factors that promote interaction with human epithelial cells . Our current understanding of H . influenzae type b and nontypable H . influenzae adhesins is reviewed in this report . In addition, models are proposed for the interrelationship of these molecules.

Am J Respir Crit Care Med, 1996 Oct, 154(4 Pt 2), S187 - 91
Aspects on the interaction of Streptococcus pneumoniae and Haemophilus influenzae with human respiratory tract mucosa; Hakansson A et al.; Haemophilus influenzae and Streptococcus pneumoniae are common causes of respiratory tract infections . H . influenzae attach to receptor epitopes in mucins and in epithelial cell membranes . Attachment is followed by an epithelial cell cytokine response . Secreted cytokines then initiate inflammation, upset the integrity of the mucosal barrier, and lead to disease . S . pneumoniae do not bind to mucins but attach to respiratory tract epithelial cells . Attachment is increased by viral infection of the epithelial cells . Unlike H . Influenzae, S . pneumoniae induce apoptosis in epithelial cells, thus disrupting the mucosal barrier . Attachment and persistence is counterbalanced by antiadhesive as well as bactericidal molecules in secretions such as human milk . These examples illustrate the balance between host defenses and microbial virulence as it has coevolved to maintain the health of the respiratory mucosa.

Chest, 1996 Oct, 110(4), 972 - 6
Detection of Mycoplasma pneumoniae by polymerase chain reaction in lung aspirates from patients with community-acquired pneumonia; Falguera M et al.; STUDY OBJECTIVE: This study was designed to evaluate the usefulness of polymerase chain reaction (PCR) to detect Mycoplasma pneumoniae DNA in samples obtained by transthoracic needle aspiration (TNA) . DESIGN: Prospective study of cases . SETTING: A university hospital in Lleida, Spain . PATIENTS: A total of 101 unselected patients, admitted between January 1993 and March 1994 in the emergency department, with a clinical and radiologic picture of community-acquired pneumonia, and without contraindications for TNA application . INTERVENTIONS: Patients were studied with conventional diagnostic techniques for community-acquired pneumonia . In addition, a sample obtained by TNA was processed by the following methods: culture in standard media, culture in selective media for Legionella, detection of capsular antigens for Streptococcus pneumoniae and Haemophilus influenzae, and detection of M pneumoniae specific genome by PCR . RESULTS: Serologic data were not available in eight patients and were excluded from this analysis . M pneumoniae PCR amplification was possible in eight cases, well correlated with serologic responses indicating current infection . Samples from ten additional patients, negative by PCR, were found to be demonstrative of recent M pneumoniae infection by serologic study . Finally, in all the remaining 75 cases, including the 59 patients for whom a different microbial diagnosis was established, M pneumoniae PCR test gave negative results . CONCLUSION: This study indicates that PCR, applied to samples obtained by TNA, appears to be a moderately sensitive and highly specific method for rapid detection of M pneumoniae lung infection.

Nucleic Acids Res, 1996 Oct 1, 24(19), 3756 - 62
Identification of new RNA modifying enzymes by iterative genome search using known modifying enzymes as probes; Gustafsson C et al.; The complete nucleotide sequences of the Haemophilus influenzae and Mycoplasma genitalium genomes and the partially sequenced Escherichia coli chromosome were analyzed to identify open reading frames (ORFs) likely to encode RNA modifying enzymes . The protein sequences of known RNA modifying enzymes from three families--m5U methyltransferases, psi synthases and 2'-O methyltransferases--were used as probes to search sequence databases for homologs . ORFs identified as homologous to the initial probes were retrieved and used as new probes against the databases in an iterative manner until no more homologous ORFs could be identified . Using this approach, we have identified two new m5U methyltransferases, seven new psi synthases and four new 2'-O methyltransferases in E . coli . Many of the ORFs found in E.coli have direct genetic counterparts (orthologs) in one or both of H.influenzae and M.genitalium . Since there is a near-complete knowledge of RNA modifications in E.coli, functional activities of the proteins encoded by the identified ORFs were proposed based on the level of conservation of the ORFs and the modified nucleotides.

Proc Natl Acad Sci U S A, 1996 Oct 1, 93(20), 11121 - 5
DNA repeats identify novel virulence genes in Haemophilus influenzae; Hood DW et al.; The whole genome sequence (1.83 Mbp) of Haemophilus influenzae strain Rd was searched to identify tandem oligonucleotide repeat sequences . Loss or gain of one or more nucleotide repeats through a recombination-independent slippage mechanism is known to mediate phase variation of surface molecules of pathogenic bacteria, including H . influenzae . This facilitates evasion of host defenses and adaptation to the varying microenvironments of the host . We reasoned that iterative nucleotides could identify novel genes relevant to microbe-host interactions . Our search of the Rd genome sequence identified 9 novel loci with multiple (range 6-36, mean 22) tandem tetranucleotide repeats . All were found to be located within putative open reading frames and included homologues of hemoglobin-binding proteins of Neisseria, a glycosyltransferase (IgtC gene product) of Neisseria, and an adhesin of Yersinia . These tetranucleotide repeat sequences were also shown to be present in two other epidemiologically different H . influenzae type b strains, although the number and distribution of repeats was different . Further characterization of the IgtC gene showed that it was involved in phenotypic switching of a lipopolysaccharide epitope and that this variable expression was associated with changes in the number of tetranucleotide repeats . Mutation of IgtC resulted in attenuated virulence of H . influenzae in an infant rat model of invasive infection . These data indicate the rapidity, economy, and completeness with which whole genome sequences can be used to investigate the biology of pathogenic bacteria.

Laryngoscope, 1996 Oct, 106(10), 1287 - 91
Unique case presentations of acute epiglottic swelling and a protocol for acute airway compromise; Parsons DS et al.; Acute epiglottitis is a well-described life-threatening disease . Since the generalized use of the Haemophilus influenzae type B (HIB) vaccine, presentations of this disorder have decreased dramatically in children . Presentations of this and other acute epiglottic swelling can vary remarkably and may easily be misdiagnosed by physicians who have little or no experience with the acutely obstructed airway . Early suspicion and a proper evaluation is mandatory to prevent a life-threatening crisis . Six patients are presented with unusual presentations of acute epiglottic swelling from differing etiologies; these include the following: case 1, recurrent epiglottitis; case 2, chronic epiglottitis; case 3, traumatic epiglottitis; case 4, caustic ingestion; and cases 5 and 6, simultaneous infection of family members . Because the incidence of acute epiglottitis is decreasing, it has become rare at most institutions . To help primary care and emergency room physicians, a formal written protocol should be available at medical facilities that might be faced with patients presenting with acute airway obstruction . An "acute airway obstruction protocol" used successfully for the last decade is offered . Use of such a written document might be life-saving for patients with impending upper airway obstruction . The otolaryngologist is a key member of the recommended multidisciplinary team.

J Bacteriol, 1996 Oct, 178(19), 5741 - 7
Cloning and characterization of two Serratia marcescens genes involved in core lipopolysaccharide biosynthesis; Guasch JF et al.; Bacteriocin 28b from Serratia marcescens binds to Escherichia coli outer membrane proteins OmpA and OmpF and to lipopolysaccharide (LPS) core (J . Enfedaque, S . Ferrer, J . F . Guasch, J . Tomas, and M . Reque, Can . J . Microbiol . 42:19-26, 1996) . A cosmid-based genomic library of S . marcescens was introduced into E . coli NM554, and clones were screened for bacteriocin 28b resistance phenotype . One clone conferring resistance to bacteriocin 28b and showing an altered LPS core mobility in polyacrylamide gel electrophoresis was found . Southern blot experiments using DNA fragments containing E . coli rfa genes as probes suggested that the recombinant cosmid contained S . marcescens genes involved in LPS core biosynthesis . Subcloning, isolation of subclones and Tn5tac1 insertion mutants, and sequencing allowed identification of two apparently cotranscribed genes . The deduced amino acid sequence from the upstream gene showed 80% amino acid identity to the KdtA protein from E . coli, suggesting that this gene codes for the 3-deoxy-manno-octulosonic acid transferase of S . marcescens . The downstream gene (kdtX) codes for a protein showing 20% amino acid identity to the Haemophilus influenzae kdtB gene product . The S . marcescens KdtX protein is unrelated to the KdtB protein of E . coli K-12 . Expression of the kdtX gene from S . marcescens in E . coli confers resistance to bacteriocin 28b.

J Bacteriol, 1996 Oct, 178(19), 5610 - 4
Mutation and Mutagenesis of thiol peroxidase of Escherichia coli and a new type of thiol peroxidase family; Cha MK et al.; A novel thioredoxin-linked thiol peroxidase (Px) from Escherichia coli has been reported previously (M . K . Cha, H . K . Kim, and I . H . Kim, J . Biol . Chem . 270:28635-28641, 1995) . In an attempt to perform physiological and biochemical characterizations of the thiol Px, a thiol Px null (tpx) mutant and a functional-residue mutant of thiol Px were produced . The tpx mutant was viable in aerobic culture but grew more slowly than the wild-type cells . The difference in growth rate became more pronounced when oxidative-stress-inducing reagents, such as peroxides and paraquat, were added to the cultures . The viability of the individual tpx mutant under oxidative stress was much lower than that of wild-type cells . tpx mutants growing aerobically respond to paraquat with a sixfold greater induction of Mn-superoxide dismutase than that of the wild-type cells . The deduced amino acid sequence of the thiol Px was found to be from 42 to 72% identical to the sequences of proteins from Haemophilus influenzae (ToxR regulon), Vibrio cholerae (ToxR regulon), and three kinds of streptococci (coaggregation-mediating adhesins), suggesting that they all belong to a new thiol Px family . Alignment of the amino acid sequences of the thiol Px family members showed that one cysteine, which corresponds to Cys-94 in E . coli thiol Px, is perfectly conserved . The substitution of serine for this cysteine residue resulted in complete loss of Px activity . These results suggest that the members of the thiol Px family, including E . coli thiol Px, have a functional cysteine residue and function in vivo as peroxidases.

Proc Natl Acad Sci U S A, 1996 Sep 17, 93(19), 10268 - 73
A minimal gene set for cellular life derived by comparison of complete bacterial genomes; Mushegian AR et al.; The recently sequenced genome of the parasitic bacterium Mycoplasma genitalium contains only 468 identified protein-coding genes that have been dubbed a minimal gene complement {Fraser, C.M., Gocayne, J.D., White, O., Adams, M.D., Clayton, R.A., et al . (1995) Science 270, 397-403} . Although the M . genitalium gene complement is indeed the smallest among known cellular life forms, there is no evidence that it is the minimal self-sufficient gene set . To derive such a set, we compared the 468 predicted M . genitalium protein sequences with the 1703 protein sequences encoded by the other completely sequenced small bacterial genome, that of Haemophilus influenzae . M . genitalium and H . influenzae belong to two ancient bacterial lineages, i.e., Gram-positive and Gram-negative bacteria, respectively . Therefore, the genes that are conserved in these two bacteria are almost certainly essential for cellular function . It is this category of genes that is most likely to approximate the minimal gene set . We found that 240 M . genitalium genes have orthologs among the genes of H . influenzae . This collection of genes falls short of comprising the minimal set as some enzymes responsible for intermediate steps in essential pathways are missing . The apparent reason for this is the phenomenon that we call nonorthologous gene displacement when the same function is fulfilled by nonorthologous proteins in two organisms . We identified 22 nonorthologous displacements and supplemented the set of orthologs with the respective M . genitalium genes . After examining the resulting list of 262 genes for possible functional redundancy and for the presence of apparently parasite-specific genes, 6 genes were removed . We suggest that the remaining 256 genes are close to the minimal gene set that is necessary and sufficient to sustain the existence of a modern-type cell . Most of the proteins encoded by the genes from the minimal set have eukaryotic or archaeal homologs but seven key proteins of DNA replication do not . We speculate that the last common ancestor of the three primary kingdoms had an RNA genome . Possibilities are explored to further reduce the minimal set to model a primitive cell that might have existed at a very early stage of life evolution.

FEMS Microbiol Lett, 1996 Sep 15, 143(1), 1 - 6
Identification of a locus involved in the utilization of iron by Actinobacillus pleuropneumoniae; Chin N et al.; The cloned afu locus of Actinobacillus pleuropneumoniae restored the ability of an Escherichia coli K-12 mutant (aroB) to grow on iron-limited media . DNA sequence analysis of the fragment showed that there are three genes designated afuA, afuB and afuC (Actinobacillus ferric uptake) that encode products similar to the SfuABC proteins of Serratia marcescens, the HitABC proteins of Haemophilus influenzae, the FbpABC proteins of Neisseria gonorrhoeae and the YfuABC proteins of Yersinia enterocolitica . The three genes encode a periplasmic iron-binding protein (AfuA), a highly hydrophobic integral cytoplasmic membrane protein with two consensus permease motifs (AfuB) and one hydrophilic peripheral cytoplasmic membrane protein with Walker ATP-binding motifs (AfuC), respectively . This system has been shown to constitute a periplasmic binding protein-dependent iron transport system in these organisms . The afuABC operon is locating approximately 200 bp upstream of apxIC gene, but transcribed in opposite direction to the ApxI-toxin genes.

Schweiz Med Wochenschr, 1996 Sep 7, 126(36), 1524 - 9
{HIV infection and splenectomy: 3 cases and literature review}; Greub G et al.; 3 splenectomized patients infected by the human immunodeficiency virus (HIV) are described . They all presented with more than 500 CD4/mm3 but, surprisingly, with a CD4 percentage below 15, positive p24 antigenemia and a CD4/CD8 ratio below 0.24 . 2 patients had repeated episodes of oropharyngeal candidiasis while their CD4 counts exceeded 800/mm3 . These episodes suggested the presence of a certain degree of immuno-suppression and prompted us to introduce anti-HIV therapy . 2 patients also presented with a pulmonary infection, due to Klebsiella pneumoniae and Haemophilus influenzae respectively . The third patient had septicemia due to Streptococcus pneumoniae type 22, despite vaccination and a CD4 count above 700/mm3 . In splenectomized HIV-infected patients the number of CD4 lymphocytes should be interpreted with caution, as this number increases after splenectomy . The CD4 percentage and CD4/CD8 ratio correlated better with the clinical stage of HIV infection and gave more valuable indications as to the degree of immunosuppression . A possible correlation between viremia and the number of CD4 lymphocytes in this subset of patients remains to be established . In HIV-infected patients, infections due to S . pneumoniae, H . influenzae, S . aureus and enteric gram-negative bacteria are frequent . After splenectomy, susceptibility to encapsulated bacteria increases even in HIV-negative patients . Early vaccination against the main strains of S . pneumoniae is essential, as vaccinal response is uncertain in patients with less than 400 CD4/mm3.

FEBS Lett, 1996 Sep 2, 392(3), 304 - 8
Purification and refolding of recombinant Haemophilus influenzae type b porin produced in Bacillus subtilis; Dahan D et al.; The major diffusion channel in the outer membrane of Haemophilus influenzae type b (Hib) is porin (341 amino acids; Mr 37 782) . The Hib porin gene was cloned and overexpressed in Bacillus subtilis . Recombinant Hib porin (Bac porin), having aggregated into inclusion bodies, was purified under denaturing conditions and subsequently refolded . To compare Bac porin that is intrinsically devoid of lipooligosaccharides versus native Hib porin, the properties of Bac porin were assessed by the following four criteria: circular dichroism spectroscopy, channel formation in planar bilayers, resistance to trypsin digestion and formation of the conformational epitope recognized by an anti-Hib porin monoclonal antibody . We conclude that in the absence of lipooligosaccharides, Bac porin was refolded into a functional form which closely resembled the structure of Hib porin.

Vaccine, 1996 Sep, 14(13), 1239 - 42
Priming with diphtheria-tetanus-pertussis vaccine enhances the response to the Haemophilus influenzae type b tetanus conjugate vaccine in infancy; Kurikka S; Haemophilus influenzae type b (Hib) capsular polysaccharide (PS) conjugated to tetanus toxoid (PRP-T) was given at 4 and 6 months of age and anti-Hib PS antibody response to the first and second dose of PRP-T was compared in groups that received diphtheria-tetanus-pertussis (DTP) vaccine either simultaneously with PRP-T (34 infants) or separately at 3, 4 and 5 months of age (49 infants) . The geometric mean anti-Hib PS antibody concentration after the first dose of PRP-T given at 4 months of age was eightfold higher if the infants primed with DTP at 3 months of age than if the first dose of DTP was given together with the first dose of PRP-T (0.81 microgram ml-1 vs 0.11 microgram ml-1) . The positive influence of DTP priming was seen also after the second dose of PRP-T given at 6 months of age (7.55 micrograms ml-1 vs 3.45 micrograms ml-1).

Vaccine, 1996 Sep, 14(13), 1219 - 22
Association of placental transfer of anti-Haemophilus influenzae type b polysaccharide antibodies with their V regions; Park MK et al.; Immunization of mothers during pregnancy may be an effective means of providing protection to infants during the first months of life against many pathogens . Previous studies have identified factors that influence the transfer of immunoglobulin across the placenta, including the time of vaccination during pregnancy and isotypes of specific immunoglobulins . By studying antibodies to Haemophilus influenzae type b polysaccharide (Hib-PS) in 26 pairs of maternal-cord sera obtained from unimmunized healthy women and 22 pairs of maternal-cord sera from women immunized with one of three different Hib vaccines, we have found that the immunoglobulin transfer is also dependent on the V region of antibodies . Anti-Hib-PS derived from the V kappa II gene "A2" was transferred about ten times more efficiently to the fetus than other anti-Hib-PS antibodies (20% vs 1-2%) . It was found that antibodies derived from the A2 V kappa gene are primarily IgG whereas other antibodies are preferentially associated with the IgM isotype . The potential association between the antibody V region with preferential placental transfer should be considered for future studies involving maternal immunization.

Diagn Microbiol Infect Dis, 1996 Sep, 26(1), 39 - 42
In vitro activity of sanfetrinem (GV104326), a new trinem antimicrobial agent, versus Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis; Doern GV et al.; Sanfetrinem, formerly GV104326, is a new trinem antimicrobial agent with extensive in vitro activity for a variety of different bacteria . The activity of sanfetrinem was determined using a broth microdilution MIC method versus a large number of clinical isolates of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis obtained in a recent 30-center United States surveillance study . The sanfetrinem MIC50 and MIC90 values for these three organism groups were 0.015 and 0.5 microgram/ml, 0.25, and 0.5 microgram/ml, and 0.015 and 0.03, respectively.

Rev Med Brux, 1996 Sep, 17(4), 250 - 2
{Vaccination of children in 1996}; Levy J; Immunization against Haemophilus influenzae b and hepatitis B during infancy, as well as the administration of a second dose of measles-mumps-rubella vaccine around the age of 12, are the significant additions brought to the childhood immunization program in recent years . The availability in the near future of the acellular pertussis vaccines illustrates the efforts made to reduce the side effects associated with the use of some vaccines . The high cost of these acellular vaccines, together with the absence of combined Haemophilus influenzae or hepatitis B vaccines, represent their current limitations.

Ann Acad Med Singapore, 1996 Sep, 25(5), 761 - 2
Haemophilus parainfluenzae infective endocarditis; Lee P et al.; Gram-negative endocarditis was uncommon in the past, accounting for 1% to 3% of cases . With the advent of antibiotics, immunosuppressive agents and narcotic abuse, the number has increased to 5% to 10% in the native valves and as high as 17% in the prosthetic valves, with Haemophilus species as the commonest aetiological agent, accounting for about 1% of the cases . We report a case of Haemophilus parainfluenzae endocarditis in a 39-year-old man who presented with heart failure and persistent fever . Echocardiography showed bi-leaflet mitral valve prolapse and severe mitral regurgitation . A small vegetation was seen at the flail anterior valve leaflet . He responded well to 4 weeks of intravenous ampicillin at 9 g/day and 2 weeks of gentamicin at 4 mg/kg/day, and subsequently underwent valve replacement.

Eur J Clin Microbiol Infect Dis, 1996 Sep, 15(9), 761 - 3
Inoculum effect and bactericidal activity of cefditoren and other antibiotics against Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis; Soriano F et al.; The inoculum effect on minimum inhibitory and minimum bactericidal concentrations of cefditoren, benzylpenicillin, ampicillin, cefotaxime, ceftriaxone, and meropenem against six clinical isolates each of Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis was studied using inocula of approximately 10(4) to 10(5) and 10(7) to 10(8) cfu/ml . Vancomycin was also studied against Streptococcus pneumoniae . The inoculum effect was observed only with benzylpenicillin and ampicillin against five of six strains of Haemophilus influenzae . All antibiotics tested were bactericidal.

Kansenshogaku Zasshi, 1996 Sep, 70(9), 947 - 54
{A clinical study of chronic lower respiratory infection with Haemophilus influenza by transtracheal aspiration}; Tsujimoto M et al.; We have performed a clinical study on chronic lower respiratory tract infection (CLRTI) with Haemophilus influenzae (H . influenzae) by transtracheal aspiration (TTA) and analyzed clinical factors of the acute exacerbation . In 40 episodes (38 cases) of H . influenzae isolated from CLRTI, monobacterial infection with H . influenzae were 21 episodes and polymicrobial infection were 19 episodes . We classified the disease into acute exacerbated (27 episodes) and stable (13 episodes) phase and the former episodes were divided into bronchitis type (7 episodes) and pneumonia type (20 episodes) . Polymicrobial infections were seen more in the pneumonia type (13 episodes) than in the bronchitis type (2 episodes) . The principal organism detected with H . influenzae were alpha-Streptococcus and Neisseria sp . in the bronchitis type and S . pneumoniae in the pneumonia type . The acute exacerbated cases were divided into the following 4 patterns; 1 . polymicrobial infection with continuous infection of P . aeruginosa, 2 . monomicrobial infection after acute upper respiratory tract infection, 3 . polymicrobial infection with S . pneumoniae after continuous infection of H . influenzae, 4 . bacterial replacement by P . aeruginosa after acute exacerbation . The results of the study suggests that polymicrobial infection is an important chronic lower respiratory tract infection when caused H . influenzae.

Changgeng Yi Xue Za Zhi, 1996 Sep, 19(3), 286 - 90
Bacterial tracheitis: a case report; Wong KS et al.; An 18-month-old girl was brought to the emergency room of Chang Gung Children's Hospital with inspiratory stridor, suprasternal retractions and imminent respiratory failure . Despite orotracheal intubation, persistent poor air-entry was noted . Flexible bronchoscopy via the endotracheal tube showed a copious amount of mucopurulent secretions in the tracheobronchial tree without any foreign bodies . With vigorous suctioning and antibiotic treatment, she had a rapid recovery . Tracheal aspirates showed a growth of Haemophilus influenzae . Cefamandole was used with good response . In conclusion, although bacterial tracheitis is an uncommon obstructive upper airway disease in children, using a bronchoscope to diagnose and to guide specific therapy can decrease the morbidity and mortality.

Onderstepoort J Vet Res, 1996 Sep, 63(3), 217 - 26
Changes in the incidences of the different serovars of Haemophilus paragallinarum in South Africa: a possible explanation for vaccination failures; Bragg RR et al.; Infectious coryza remains an important disease in the poultry industry despite the long-term and widespread use of vaccines against its causative agent, Haemophilus paragallinarum, in South Africa . In order to detect antigenic changes between populations of H . paragallinarum isolated before the use of vaccines against infectious coryza in this country, and field isolates obtained after the introduction of infectious coryza vaccines, 106 different NAD-dependent isolates (of which 93 were identified as H . paragallinarum) from 63 different farms, and dating from 1972 to March 1995, were identified by means of rabbit antisera against serogroups A, B and C . Serogroup C isolates show weaker cross-protection, requiring the further subdivision of this serogroup into its four different serovars . The percentages of the different serovars obtained in the 1970s, confirmed previously published data on South African isolates . A tendency towards a decrease in the number of serogroup A and serovar C-2 isolates, and an increase in the percentage of serovar C-3 isolates, was noted among isolates of the 1980s . These changes were markedly enhanced in the isolates obtained from 1990 to March 1995 . The percentage of serogroup A isolates decreased significantly from 34% in the 1970s to only 5% in the 1990s, and that of serovar C-2 isolates, from 31-18%, while the abundance of serovar C-3 isolates increased significantly from 31% in the 1970s to 73% in the 1990s . Serogroup B remained more or less constant and never reached more than 10% of the population . These results indicate the need for the incorporation of serovar C-3 in a vaccine for use in South Africa, particularly in those areas of the country from which isolates were collected during this study . Some of the NAD-dependent isolates obtained from poultry in South Africa between 1970 and 1995, were biochemically identified as Pasteurella avium and P . volantium . As H . avium has been subdivided and reclassified into the genus Pasteurella, this represents the first report of the identification of P . avium and P . volantium in South Africa.

Acta Otolaryngol, 1996 Sep, 116(5), 766 - 73
Persistence of nontypeable Haemophilus influenzae in adenoid macrophages: a putative colonization mechanism; Forsgren J et al.; That nontypeable H . influenzae (NTHI) can reside intracellularly in human adenoid tissue has been suggested by use of in situ hybridization of a fluorescein labelled 16S rRNA-targeted oligonucleotide probe (FISH) . Adenoid tissues from 43 children operated on in a clinically infection-free interval were investigated . FISH revealed H . influenzae in macrophage-like cells, located subepithelially in the crypts in all 43 adenoids . Furthermore, H . influenzae was detected in 22/22 adenoids using immunohistochemistry with the monoclonal antibody MAHI-3 recognizing a conserved H . influenzae LPS inner-core region . FISH and staining with monoclonal antibodies against immunophenotypic markers were performed simultaneously in order to characterize the cellular interrelations in this microenvironment . The findings of widespread presence of H . influenzae in cells of which some strongly expressed the CD14 marker of the monocyte/macrophage lineage may correspond to an important aspect of the colonization mechanisms whereby NTHI persists in the nasopharynx of children.

Ann Trop Paediatr, 1996 Sep, 16(3), 193 - 8
Dexamethasone adjunctive therapy for bacterial meningitis in children: a retrospective study in Brazil; Macaluso A et al.; The clinical records of 87 children with bacterial meningitis treated with antibiotics (group 1) and of 92 treated with antibiotics plus dexamethasone (group 2), admitted to the Instituto Materno Infantil de Pernambuco, Recife, Brazil over 2 consecutive years (1991 and 1992), were analysed . There were no significant differences between treatment groups regarding characteristics on admission except that group 1 were younger . The overall case fatality rate was 19%, with 14% in group 2 and 24% in group 1 (p = 0.09) . Rate of discharge without sequelae was 70% in the steroid-treated children and 56% in children treated with antibiotics alone (p = 0.07) . Among children aged 6-59 months, those treated with dexamethasone compared with those treated with antibiotics alone had a better case fatality rate (11% vs 25%; p = 0.05) and a better rate of discharge without sequelae (73% vs 52%; p = 0.02) . Among the cases with a CSF culture positive for Haemophilus influenzae, 77% were discharged without sequelae in group 2 compared with 51% in group 1 (p = 0.03) . The addition of dexamethasone to standard antibiotic treatment improves the outcome of children between 6 and 59 months of age admitted to hospital with a diagnosis of bacterial meningitis.

J Antimicrob Chemother, 1996 Sep, 38(3), 363 - 86
Susceptibility testing of Haemophilus influenzae--an international collaborative study in quality assessment; Yeo SF et al.; In order to compare the prevalence of antibiotic resistance in different geographical areas, it is necessary to ensure that agreement is achieved between laboratories on the assignment of strains to 'susceptible' and 'resistant' categories . An international quality assessment study, involving 15 laboratories in eight countries, was performed to investigate the standard of performance of the susceptibility testing of Haemophilus influenzae . One hundred and fifty strains of H . influenzae were distributed from the London Hospital Medical College (LHMC) to all laboratories who were asked to test the susceptibility of the strains to ampicillin, chloramphenicol, tetracycline, trimethoprim, cephalosporins and ciprofloxacin . Laboratories were also asked to provide the details of methodology to test the susceptibility . Significant discrepancy between the LHMC and the participating laboratories appeared in the detection of resistance to ampicillin (especially beta-lactamase-negative strains resistant to ampicillin) as well as the assignment of susceptibility and resistance to chloramphenicol, tetracycline and trimethoprim . Often these reflected the use of inappropriate breakpoints which led to erroneous assignment of susceptibility . Other variations including disc content, medium and supplement, inoculum as well as failure to measure zone sizes properly also led to some repeating anomalies.

Mol Microbiol, 1996 Sep, 21(5), 941 - 52
Regulation of competence development and sugar utilization in Haemophilus influenzae Rd by a phosphoenolpyruvate:fructose phosphotransferase system; Macfadyen LP et al.; Changes in intracellular cAMP concentration play important roles in Haemophilus influenzae, regulating both sugar utilization and competence for natural transformation . In enteric bacteria, cAMP levels are controlled by the phosphoenolpyruvate:glycose phosphotransferase system (PTS) in response to changes in availability of the preferred sugars it transports . We have demonstrated the existence of a simple PTS in H . influenzae by several methods . We have cloned the H . influenzae ptsI gene, encoding PTS Enzyme I; genome analysis locates it in a pts operon structurally homologous to those of enteric bacteria . In vitro phosphorylation assays confirmed the presence of functional PTS components . A ptsI null mutation reduced fructose uptake to 1% of the wild-type rate, and abolished fructose fermentation even when exogenous cAMP was provided . The ptsI mutation also prevented fermentation of ribose and galactose, but utilization of these cAMP-dependent sugars was restored by addition of cAMP . In wild-type cells the non-metabolizable fructose analogue xylitol prevented fermentation of these sugars, confirming that the fructose PTS regulates cAMP levels . Development of competence under standard inducing conditions was reduced 250-fold by the ptsI mutation, unless cells were provided with exogenous cAMP . Competence is thus shown to be under direct nutritional control by a fructose-specific PTS.

Semin Respir Infect, 1996 Sep, 11(3), 139 - 47
Antimicrobial therapy of pneumonia in infants and children; Harris JA; To provide optimal management for the child with community-acquired pneumonia, the clinician must take multiple factors into consideration . The etiology of pneumonia is difficult to determine and initial choice of therapy is based on the frequency of pathogens in various age groups, local antibiotic resistance patterns of the organisms, clinical presentation, and epidemiological data . Streptococcus pneumoniae and Haemophilus influenzae remain the most common bacterial pathogens outside the newborn period . Increasing numbers of multidrug-resistant strains of S pneumoniae in the United States and Europe, the decline in H influenzae type b because of current vaccination strategies, and increasing recognition of nontypeable H influenzae as etiologic agents of pneumonia have prompted reconsideration of the drug of choice . Amoxicillin and its derivatives or oral cephalosporins are the drugs of choice for initial therapy for mild to moderate disease . For severe disease or if beta-lactamase producing organisms are a concern, extended spectrum cephalosporins are indicated . Pneumococcal pneumonia unresponsive to penicillin therapy may warrant the use of extended spectrum cephalosporins or vancomycin . For older children in whom mycoplasma is a significant cause of pneumonia, the new macrolides have provided additional options for the clinician . Azithromycin and clarithromycin are efficacious, well tolerated, and require less frequent dosing intervals . The introduction of ceftriaxone, a third-generation cephalosporin with a broad spectrum of activity and prolonged half-life, allows once-a-day intramuscular therapy that can be administered on an outpatient basis . With the availability of parenteral outpatient therapy, hospital admission is no longer required for the treatment of most cases of serious community-acquired pneumonia.

Eur Respir J, 1996 Sep, 9(9), 1913 - 22
Bacterial-induced release of inflammatory mediators by bronchial epithelial cells; Khair OA et al.; This review focuses on bacterial induction and release of inflammatory cytokines and adhesion molecules by human bronchial epithelial cells, with special reference to Haemophilus influenzae, a pathogen commonly associated with chronic bronchitis . Studies investigating the mechanisms underlying bacterial colonization of the airways and bacterial-induced chronic airway inflammation have suggested that these are likely to involve localization of bacteria to the site(s) of infection in the respiratory tract and induction of a local airway inflammation resulting in the initiation of epithelial damage . We have hypothesized that the gross airway epithelial damage observed in chronic infective lung disease is an indirect consequence of proteolytic enzymes and toxic oxygen radicals generated by large numbers of neutrophils infiltrating the airways . Furthermore, the infiltration and activation of the neutrophils is a consequence of increased release of proinflammatory mediators from the host respiratory epithelium, induced by bacterial products, such as endotoxin . This hypothesis is based on studies which have demonstrated that the concentrations of circulating cytokines, such as interleukin (IL)-8 and tumour necrosis factor-alpha (TNF-alpha), which have profound effects on neutrophil activity, are increased in endotoxaemia and that airway epithelial cells are a rich source of these cytokines . Support for this hypothesis is provided by studies of cultured human bronchial epithelial cells incubated either in the absence or presence of purified endotoxin preparations from nontypable and type b H . influenzae strains which have demonstrated that these endotoxins lead to significantly increased expression and/or release of proinflammatory mediators, including IL-6, IL-8, TNF-alpha and intercellular adhesion molecule-1 (ICAM-1) . Treatment of the cells with steroids can downregulate the expression and/or release of these inflammatory mediators . Additionally, these studies have demonstrated that culture medium collected from endotoxin-treated cultures, 24 h after treatment, significantly increases neutrophil chemotaxis and adhesion to human endothelial cells in vitro.

Antimicrob Agents Chemother, 1996 Sep, 40(9), 2131 - 6
Genetic characterization of trimethoprim resistance in Haemophilus influenzae; de Groot R et al.; We previously demonstrated that trimethoprim (Tmp) resistance in Haemophilus influenzae is mediated by chromosomally encoded dihydrofolate reductase (DHFR) with a modified primary structure and distinct kinetic properties . To gain insight into the relationship of the DHFR structure and the level of Tmp resistance that it confers on the host bacterium, we cloned and characterized the folH genes of one Tmp-susceptible and two Tmp-resistant H . influenzae strains . Differences were observed between Tmp-susceptible and Tmp-resistant isolates both in the promoter region and in the coding sequences . The effect of differences between H . influenzae folH genes on Tmp susceptibility was investigated in Escherichia coli . Various folH gene hybrids were constructed, and their influence on Tmp susceptibility was determined . Resistance in E . coli mediated by folH from H . influenzae strain R1047 was associated with alterations in the promoter and the central part of folH . In contrast, the E . coli Tmp resistance phenotype associated with the folH gene of H . influenzae R1042 was characterized by alterations in one or more of three amino acid residues at the C-terminal part of the protein . These data indicate that Tmp resistance is not only related to alterations in the promoter region of the folH gene and the Tmp binding domains at the N-terminal and central part of DHFR . Alterations in the C-terminal part may also cause Tmp resistance, probably as a result of a change in secondary structure and the subsequent loss of Tmp binding affinity.

Pediatr Infect Dis J, 1996 Sep, 15(9 Suppl), S30 - 7
Properties of azithromycin that enhance the potential for compliance in children with upper respiratory tract infections; Powers JL; BACKGROUND: Azithromycin, the prototypical azalide antibiotic, has a wide spectrum of activity that is characterized by resistance to beta-lactamase-producing microbes and efficacy against Gram-positive and Gram-negative pathogens, including Haemophilus influenzae . Tissue-directed pharmacokinetics include tissue concentrations up to 100-fold higher than those in plasma and a tissue half-life of up to 4 days . Pharmacokinetics of azithromycin permits a reduction in dosage frequency and duration while maintaining efficacy comparable to that of conventional 7- to 10-day three or four times daily regimens . Dosage interval, duration of treatment, side effects and palatability can affect compliance and thus clinical outcome . Compliance among children is important in light of the high incidence of community-acquired infections such as otitis media and streptococcal pharyngitis . OBJECTIVE: To compare the flavor, taste acceptability and color preference of oral antibiotic suspensions given to children . METHODS: The taste and acceptability of the oral suspension form of azithromycin vs . cefixime, cefpodoxime proxetil, cefprozil, clarithromycin or loracarbef were rated by children during blinded taste tests and with acceptability/ preference questionnaires . RESULTS: Analysis of the mean acceptability/ preference rating from 769 children demonstrated that the flavor of azithromycin was rated significantly higher than that of cefpodoxime (4.3 vs . 2.8), cefprozil (4.0 vs . 3.4) and clarithromycin (4.3 vs . 2.7) and was comparable to that of cefixime (4.0 vs . 4.2) and loracarbef (4.4 vs . 4.5) . A greater percentage of children preferred the taste of azithromycin to that of cefpodoxime (90.0% vs . 5.2%), cefprozil (63.0% vs . 33.1%) and clarithromycin (89.0% vs . 11.0%) . The taste of azithromycin was not preferred to that of cefixime (39.0% vs . 53.9%) or loracarbef (36% vs . 58.5%) . CONCLUSIONS: The efficacy and safety of azithromycin in otitis media and streptococcal pharyngitis, the simple dosing regimen and a highly palatable oral suspension formulation should increase compliance among pediatric patients and thereby improve clinical outcomes.

Pediatr Infect Dis J, 1996 Sep, 15(9 Suppl), S15 - 9
A multicenter, open label trial of azithromycin vs . amoxicillin/ clavulanate for the management of acute otitis media in children; Aronovitz G; OBJECTIVE: This randomized, open label study compared the efficacy and safety of a 5-day course of once daily azithromycin to those of a 10-day course of three times daily amoxicillin/ clavulanate . METHODS: One hundred sixty-nine children with confirmed acute bacterial otitis media were randomized to treatment with 10 mg/kg of azithromycin oral suspension on Day 1, followed by 5 mg/kg once daily for the next 4 days or 40 mg/kg/ day of amoxicillin/clavulanate suspension in three divided doses for 10 days . Clinical efficacy and safety were evaluated on Days 11 and 30 . RESULTS: Analysis of evaluable patients 11 days after the start of treatment demonstrated that 87.8% of patients treated with azithromycin and 100.0% of the patients treated with amoxicillin/ clavulanate were cured or improved . Presumed bacteriologic eradication of the baseline pathogens Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pyogenes was comparable in the two groups: 87.8% in patients treated with azithromycin; and 100.0% in patients receiving amoxicillin/clavulanate . At Day 30, 82.2% of patients treated with azithromycin and 80.0% of patients treated with amoxicillin/clavulanate were cured or improved . Presumed bacteriologic eradication of the base-line pathogens was similar in the two groups: 82.2% in patients treated with azithromycin; and 81.1% in patients receiving amoxicillin/clavulanate . Relapses occurred in 5.1% of patients receiving azithromycin and 21.1% of patients taking amoxicillin/clavulanate (P = 0.047) . Treatment-related side effects occurred in 3.5% of the azithromycin patients compared with 31.0% of amoxicillin/clavulanate patients (P < 0.001) . CONCLUSIONS: Azithromycin was comparable to amoxicillin/clavulanate in achieving clinical cure or improvement and presumed eradication of baseline pathogens in pediatric patients with acute otitis media . Azithromycin was significantly better tolerated and was associated with fewer relapses than seen after amoxicillin/clavulanate therapy.

Pediatr Infect Dis J, 1996 Sep, 15(9 Suppl), S10 - 4
A multicenter, open label trial of azithromycin for the treatment of children with acute otitis media; McCarty J; OBJECTIVE: In this multicenter, open label trial the investigators evaluated the efficacy and safety of azithromycin suspension administered once daily for 5 days for the treatment of clinically and bacteriologically established acute otitis media . METHODS: Two hundred eligible children with acute otitis media from 10 US centers were treated with 10 mg/kg of azithromycin oral suspension on Day 1, followed by 5 mg/kg once daily for the next 4 days . Tympanocentesis and subsequent culture of middle ear effusion were performed at baseline . Clinical efficacy was evaluated on Days 6, 11 and 30 . RESULTS: Analysis of clinical efficacy in evaluable patients 11 days after the initiation of therapy showed that the rate of satisfactory responses (cured or improved) ranged from 79.6 to 82.4% in patients infected with Streptococcus pneumoniae, Haemophilus influenzae, or Moraxella catarrhalis . Satisfactory clinical response at Day 30 was reported in 70% of evaluable patients, and eradication of S . pneumoniae, H . influenzae and M . catarrhalis was presumed in 64 to 73% . Relapses occurred in 14% of the evaluable patients . Among the treated patients 8.5% reported mild or moderate side effects . CONCLUSION: Azithromycin is an effective, safe and well-tolerated treatment for children with acute otitis media.

Pediatr Infect Dis J, 1996 Sep, 15(9 Suppl), S3 - 9
Incidence of antibiotic-resistant Streptococcus pneumoniae and beta-lactamase-positive Haemophilus influenzae in clinical isolates from patients with otitis media; McLinn S et al.; BACKGROUND: The prevalence of penicillin-resistant Streptococcus pneumoniae and beta-lactamase-producing Haemophilus influenzae in otitis media infections is increasing; emergence of these pathogens has complicated treatment . OBJECTIVES: To evaluate the incidence of penicillin resistance and the in vitro activity of amoxicillin/clavulanate, cefaclor, loracarbef, cefixime, trimethoprim/sulfamethoxazole, azithromycin and clarithromycin in S . pneumoniae isolates . The in vitro activity of azithromycin, clarithromycin and cefaclor was also evaluated in beta-lactamase-positive and -negative isolates of H . influenzae . METHODS: Bacterial isolates of S . pneumoniae and H . influenzae were obtained by tympanocentesis and subsequent culture of middle ear effusion from children with acute otitis media enrolled in a multicenter trial . Susceptibility to test agents was assessed by disk diffusion and broth dilution techniques with criteria established by the National Committee for Clinical Laboratory Standards . RESULTS: Nineteen (31%) of the 61 S . pneumoniae isolates were resistant to penicillin . A significantly lower percentage of the S . pneumoniae isolates were resistant to azithromycin (16%) and clarithromycin (11%) than to penicillin, amoxicillin/ clavulanate, cefaclor, loracarbef or cefixime (31% in all cases) . Azithromycin was also more active than cefaclor and significantly more active than clarithromycin against the 55 H . influenzae isolates . CONCLUSIONS: The susceptibility of resistant and nonresistant strains of S . pneumoniae to azithromycin and clarithromycin and of isolates of H . influenzae to azithromycin, coupled with penetration of azithromycin into the middle ear, may provide a significant advantage in the treatment of otitis media.

Microbiology, 1996 Sep, 142 ( Pt 9), 2587 - 94
Organization of the Haemophilus ducreyi 35000 chromosome; Hobbs MM et al.; A physical and rudimentary genetic map of the Haemophilus ducreyi strain 35000 genome was constructed . Pulsed-field gel electrophoresis was used to separate restriction fragments of H . ducreyi DNA digested with Sfil, I-Ceul, or Sfil plus I-Ceul . The sizes of the fragments were determined, and the circular chromosome was estimated to be 1757 kbp . The six I-Ceul fragments and four Sfil fragments were ordered into macrorestriction maps using Southern blot hybridization with random H . ducreyi clones as probes . It was shown that both H . ducreyi and the distantly related Haemophilus influenzae have six rrn operons marked by the locations of the I-Ceul sites . However, the two species displayed distinct I-Ceul restriction patterns . A second H . ducreyi strain, CIP542, displayed an identical I-Ceul pattern to that of H . ducreyi 35000, but Sfil digests of teh two strains were distinct . The orientation of the six rrn operons was determined and thirteen identified H . ducreyi genes positioned on the map of strain 35000.

Microbiology, 1996 Sep, 142 ( Pt 9), 2449 - 52
Molecular analysis of a new insertion sequence from Actinobacillus (Haemophilus) actinomycetemcomitans FDC Y4; Hayashida H et al.; We have found a new insertion sequence (IS), designated ISAa1, downstream of the S10 operon in Actinobacillus (Haemophilus) actinomycetemcomitans FDC Y4 . ISAa1, the first IS element characterized in this organism, is 705 bp long and lacks terminal inverted repeats . This element displayed significant homology with IS200 . Hybridization patterns of genomic DNA of seven A . actinomycetemcomitans strains with an internal ISAa1 probe varied depending on the serotypes, suggesting that ISAa1 might be a useful tool for epidemiological studies.

Clin Exp Immunol, 1996 Sep, 105(3), 537 - 43
Anti-inflammatory properties of human serum IgA: induction of IL-1 receptor antagonist and Fc alpha R (CD89)-mediated down-regulation of tumour necrosis factor-alpha (TNF-alpha) and IL-6 in human monocytes; Wolf HM et al.; A deregulated expression and/or release of large amounts of inflammatory cytokines such as IL-1 and TNF-alpha accounts for most pathophysiological events in a variety of systemic inflammatory diseases, the effect being mediated by the interaction of these cytokines with their respective receptors . IL-1 receptor antagonist (IL-1Ra), mainly produced by monocytes/macrophages, is an inhibitor of IL-1 activity . The present study shows that human serum IgA induces significant IL-1Ra release in human peripheral blood mononuclear cells and adherent monocytes . IgA induced higher levels of IL-1Ra than Haemophilus influenzae type b (Hib) expressing lipopolysaccharide (LPS), purified LPS or phorbol myristate acetate (PMA), without induction of IL-1 beta release, and even inhibited LPS-induced IL-1 beta release . Induction of IL-1Ra by IgA could be detected both at the mRNA and protein levels in resting and activated monocytes . Ligation of Fc alpha R with MoAb My-43 or treatment with human serum IgA induced protein tyrosine phosphorylation in human monocytes, and herbimycin A, a specific inhibitor of protein tyrosine kinase activity, inhibited IgA-induced IL-1Ra production, suggesting that Fc alpha R-mediated induction of tyrosine phosphorylation is required for the IgA-induced stimulation of IL-1Ra release . In addition, triggering of Fc alpha R with MoAb specifically down-regulated TNF-alpha and IL-6 release in human monocytes activated with Hib . By the induction of IL-1Ra and down-regulation of the release of inflammatory cytokines such as IL-1 beta, TNF-alpha and IL-6, interaction of IgA with human monocytes may actively contribute to the regulation of the inflammatory response.

Clin Exp Immunol, 1996 Sep, 105(3), 422 - 8
Expression of Haemophilus influenzae type b idiotype 1 on naturally acquired antibodies; Ulanova M et al.; The Chinese population in Hong Kong has a low incidence of invasive Haemophilus influenzae type b(Hib) disease, as well as carriage of the microorganism . Likely stimuli for the natural antibodies to Hib, which might protect against Hib infection, are cross-reactive antigens of bacteria like Escherichia coli K 100 . Our aim was to determine the isotype and idiotype distribution and cross-reactivity of natural antibodies against Hib capsular polysaccharide (CP) in healthy Hong Kong Chinese . Titration of 20 sera by ELISA showed IgG antibodies reacting with Hib CP in all individuals . The antibodies were mainly IgG2, and their avidity index ranged widely . Isoelectric focusing (IEF) combined with immunoblotting showed patterns of IgG2 antibody clones against the CP of Hib and E . coli K 100 which were similar in 10 cases . Absorption with Hib CP only eliminated some bands in two sera . Absorption with K 100 CP did not remove any anti-Hib CP bands . In three sera additional clones of antibodies reacting to K 100 CP only, disappeared after absorption with this CP . Spectrotypic analyses of IgG antibodies reacting with anti-Hib idiotype 1 (Id-1) revealed stronger IEF patterns with bands in differing locations compared with anti-Hib CP antibodies . The strong reactivity of serum IgG, IgA and IgM antibodies with monoclonal anti-Hib Id-1 was confirmed by ELISA . This reactivity was not abolished after absorption of the sera with either Hib CP, or K 100 CP . The data indicate a high prevalence of Id-1 among Hong Kong Chinese . However, only one individual had Id-1 antibodies specific for Hib CP, judging from absorption experiments . Others had much lower activity of Id-1 anti-Hib CP antibodies compared with the total IgG Id-1, suggesting that Hong Kong subjects have Id-1-positive antibodies in their serum which are not specific for Hib CP . This is consistent with the nature of Id-1, which is a marker of A2VL region usage rather than a marker of a Hib CP paratope . We suggest that natural antibodies reacting with Hib CP in healthy Hong Kong Chinese are the product of exposure to some cross-reactive antigen(s), different from both Hib and E . coli K 100 CP.

Endocrinology, 1996 Sep, 137(9), 4037 - 45
Constitutive nuclear localization of Janus kinases 1 and 2; Lobie PE et al.; Both GH and the GH receptor have been reported to undergo rapid nuclear translocation . Janus kinases (JAK) 1 and 2 have been implicated in GH receptor signaling, and both of these kinases are phosphorylated by GH stimulation . In this report, we have investigated the subcellular distribution of JAK1 and JAK2 . Both JAK1 and JAK2 exhibit a nucleocytoplasmic distribution by immunocytochemistry in unstimulated serum deprived CHO cells stably transfected with rat GH receptor complementary DNA (cDNA) . The nucleocytoplasmic localization of JAK2 was verified by immunogold electron microscopy in both rat liver hepatocytes and CHO cells stably transfected with rat GH receptor cDNA . Nucleocytoplasmic localization of JAK2 was also verified by transient tranfection of CHO cells with a Haemophilus influenzae haemagglutinin (HA) epitope tagged JAK2 expression plasmid and subsequent localization of HA immunoreactivity . Western blot analysis of purified nuclear extracts revealed the presence of immunoreactive JAK1 at 130 kDa and immunoreactive JAK2 at 128 kDa . No change in the nuclear content of JAK1 or JAK2 was observed upon ligand stimulation of GH receptor cDNA transfected cells with 100 nM human GH for 5, 10, 15, 30, or 60 min . GH stimulation caused, however, the appearance of tyrosine phosphorylated 42- and 44-kDa proteins as well as tyrosine phosphorylated JAK2 in the nucleus . The constitutive nuclear localization of the Janus Kinases is suggestive of a novel nuclear role for JAK family members, in addition to their described cytosolic function and presents an interesting challenge to the subcellular site of hormone action.

J Bacteriol, 1996 Sep, 178(17), 5086 - 91
The C terminus of the AddA subunit of the Bacillus subtilis ATP-dependent DNase is required for the ATP-dependent exonuclease activity but not for the helicase activity; Haijema BJ et al.; Comparison of subunit AddA of the Bacillus subtilis AddAB enzyme, subunit RecB of the Escherichia coli RecBCD enzyme, and subunit RecB of the Haemophilus influenzae RecBCD enzyme revealed several regions of homology . Whereas the first seven regions are common among helicases, the two C-terminally located regions are unique for RecB of E . coli and H . influenzae and AddA . Deletion of the C-terminal region resulted in the production of an enzyme which showed moderately impaired levels of ATP-dependent helicase activity, whereas the ATP-dependent exonuclease activity was completely destroyed . The mutant enzyme was almost completely capable of complementing E . coli recBCD and B . subtilis addAB strains with respect to DNA repair and homologous recombination . These results strongly suggest that at least part of the C-terminal region of the AddA protein is indispensable for exonuclease activity and that, in contrast to the exonuclease activity, the helicase activity of the addAB gene product is important for DNA repair and homologous recombination.

J Bacteriol, 1996 Sep, 178(17), 5071 - 9
Three transport systems for the osmoprotectant glycine betaine operate in Bacillus subtilis: characterization of OpuD; Kappes RM et al.; The accumulation of the osmoprotectant glycine betaine from exogenous sources provides a high degree of osmotic tolerance to Bacillus subtilis . We have identified, through functional complementation of an Escherichia coli mutant defective in glycine betaine uptake, a new glycine betaine transport system from B . subtilis . The DNA sequence of a 2,310-bp segment of the cloned region revealed a single gene (opuD) whose product (OpuD) was essential for glycine betaine uptake and osmoprotection in E . coli . The opuD gene encodes a hydrophobic 56.13-kDa protein (512 amino acid residues) . OpuD shows a significant degree of sequence identity to the choline transporter BetT and the carnitine transporter CaiT from E . coli and a BetT-like protein from Haemophilus influenzae . These membrane proteins form a family of transporters involved in the uptake of trimethylammonium compounds . The OpuD-mediated glycine betaine transport activity in B . subtilis is controlled by the environmental osmolarity . High osmolarity stimulates de novo synthesis of OpuD and activates preexisting OpuD proteins to achieve maximal glycine betaine uptake activity . An opuD mutant was constructed by marker replacement, and the OpuD-mediated glycine betaine uptake activity was compared with that of the previously identified multicomponent OpuA and OpuC (ProU) glycine betaine uptake systems . In addition, a set of mutants was constructed, each of which synthesized only one of the three glycine betaine uptake systems . These mutants were used to determine the kinetic parameters for glycine betaine transport through OpuA, OpuC, and OpuD . Each of these uptake systems shows high substrate affinity, with Km values in the low micromolar range, which should allow B . subtilis to efficiently acquire the osmoprotectant from the environment . The systems differed in their contribution to the overall glycine betaine accumulation and osmoprotection . A triple opuA, opuC, and opuD mutant strain was isolated, and it showed no glycine betaine uptake activity, demonstrating that three transport systems for this osmoprotectant operate in B . subtilis.

Infect Immun, 1996 Sep, 64(9), 3703 - 12
Identification of an outer membrane protein involved in utilization of hemoglobin-haptoglobin complexes by nontypeable Haemophilus influenzae; Maciver I et al.; A recombinant plasmid containing a 6.5-kb fragment of nontypeable Haemophilus influenzae (NTHI) chromosomal DNA was shown to confer a hemoglobin-haptoglobin-binding phenotype on Escherichia coli . Use of a mini-Tn10kan transposon for random insertion mutagenesis of this recombinant plasmid allowed localization of the NTHI DNA responsible for this hemoglobin-haptoglobin-binding phenotype to a 3.5-kb PstI-XhoI fragment within the 6.5-kb NTHI DNA insert . When this mutagenized NTHI DNA fragment was used to transform the wild-type NTHI strain, the resultant kanamycin-resistant mutant exhibited significantly decreased abilities to bind hemoglobin-haptoglobin and utilize it as a source of heme for aerobic growth in vitro . This mutant also lacked expression of a 115-kDa outer membrane protein that was present in the wild-type parent strain . Transformation of this mutant with wild-type NTHI chromosomal DNA restored the abilities to bind and utilize hemoglobin-haptoglobin and to express the 115-kDa outer membrane protein . Nucleotide sequence analysis of the relevant NTHI DNA revealed the presence of a gene, designated hhuA, that encoded a predicted 117,145-Da protein . The HhuA protein exhibited features typical of a TonB-dependent outer membrane receptor and had significant identity with the hemoglobin receptors of both Haemophilus ducreyi and Neisseria meningitidis.

Infect Immun, 1996 Sep, 64(9), 3666 - 72
DNA sequence analysis and restriction fragment length polymorphisms of the P1 gene of Haemophilus influenzae biogroup aegyptius associated with Brazilian purpuric fever; Reed RB et al.; Brazilian purpuric fever (BPF) is a fulminant pediatric disease caused by specific strains of Haemophilus influenzae biogroup aegyptius . A conserved epitope on the P1 protein of strains of H . influenzae biogroup aegyptius is seen on most virulent isolates . The P1 protein from a Brazilian case-clone strain of H . influenzae biogroup aegyptius was analyzed by cloning and sequencing the gene . Three major variable regions are present within the P1 gene of the BPF clone in an architecture similar to that of the previously sequenced P1 genes from H . influenzae . The DNA sequence data of the P1 gene provided information for restriction fragment length polymorphism analyses among strains of H . influenzae biogroup aegyptius . Using PCR for amplification of the P1 gene, we found that AlwI restriction of this gene allowed for a highly accurate segregation of virulent strains of H . influenzae biogroup aegyptius associated with BPF . The strong association of virulent phenotypes with specific AlwI restriction patterns of the P1 gene provides a basis for the convenient and accurate identification of strains of H . influenzae biogroup aegyptius which cause BPF.

FEBS Lett, 1996 Aug 26, 392(2), 184 - 8
The monofunctional glycosyltransferase of Escherichia coli is a member of a new class of peptidoglycan-synthesising enzymes; Di Berardino M et al.; Using conserved fingerprints in the glycosyltransferase (GTase) domain of high-molecular-weight penicillin-binding proteins (PBP), a gene (mgt) encoding a putative monofunctional glycosyltransferase has been identified in Haemophilus influenzae and in other bacteria} species . Here we report the cloning of the homologous Escherichia coli gene and show that the solubilised membrane fraction of E . coli cells overexpressing the mgt gene contain a significantly increased peptidoglycan synthesis activity . In contrast to the high-molecular-weight PBPs, this activity is not inhibited by Flavomycin.

FEMS Microbiol Lett, 1996 Aug 15, 142(1), 11 - 7
Occurrence of {copper, zinc}-cofactored superoxide dismutase in Pasteurella haemolytica and its serotype distribution; Lainson FA et al.; Fifty-two ovine strains of Pasteurella haemolytica and P . trehalosi representing serotypes 1-16 were examined for the presence of {copper, zinc}superoxide dismutase DNA sequences . This was done using a combination of polymerase chain reaction with degenerate primers based on the sequence of the {Cu,Zn}superoxide dismutase gene (sodC) in related species and Southern hybridization using a fragment of sodC from P . haemolytica A2 serotype as a probe . Both detection methods identified a fragment of the sodC gene in 9/9 strains of P . haemolytica serotype 2 examined and in 5/8 strains of serotype 7 . No evidence of this gene was found in any other serotype of P . haemolytica or in any P . trehalosi serotype . Comparison of DNA sequence showed near identity between sodC from the A2 and A7 serotypes of P . haemolytica and substantial similarity (70%) to sodC previously sequenced in P . multocida, Haemophilus parainfluenzae and H . influenzae . Analysis by gel electrophoresis of the superoxide dismutase activity present in cell lysates showed that one or more superoxide dismutase is present in all serotypes . However, cyanide-inhibitable activity, corresponding to {Cu,Zn}superoxide dismutase, was detected only in those strains of serotypes A2 and A7 which showed evidence of the sodC gene fragment.

Pulm Pharmacol, 1996 Aug, 9(4), 223 - 30
The effect of bacterial toxins on levels of intracellular adenosine nucleotides and human ciliary beat frequency; Kanthakumar K et al.; Toxins that slow ciliary beat are virulence determinants of bacteria that infect or invade ciliated epithelial surfaces . We have previously shown that the effect of the Pseudomonas aeruginosa toxin pyocyanin on ciliary beat is associated with a fall in intracellular cAMP and ATP . We have now investigated whether reduction in intracellular adenosine nucleotides might be a common mechanism of action of other bacterial toxins which slow ciliary beat . Two other P . aeruginosa toxins, 1-hydroxyphenazine (1-HP) and rhamnolipid, and two Haemophilus influenzae fractions produced by gel filtration of broth cultures were tested . The effect on human nasal epithelium ciliary beat frequency (CBF), and intracellular cAMP and ATP were measured, and the effect of two pharmacological agents, dibutyryl cAMP and salmeterol, on these changes was assessed . 1-HP, rhamnolipid and the two H . influenzae fractions slowed CBF before there was significant release of lactate dehydrogenase from the cells . The toxins also caused a fall in intracellular cAMP and ATP . Dibutyryl cAMP and salmeterol at the concentrations used do not increase baseline CBF, but diminished the fall in CBF and intracellular adenosine nucleotides . The cAMP and ATP levels in these studies were combined with those previously obtained with pyocyanin . there was a good correlation between cAMP and ATP levels and CBF . Bacterial toxins which slow CBF may act by causing a fall in intracellular adenosine nucleotides, and agents which stimulate cAMP may prevent toxin-induced slowing of ciliary beat.

Jpn J Antibiot, 1996 Aug, 49(8), 800 - 7
{Clinical evaluation of sulbactam/cefoperazone for lower respiratory tract infections . Correlation between the efficacy of sulbactam/cefoperazone and beta-lactamase}; Koga H et al.; The efficacy and safety of sulbactam/cefoperazone (SBT/CPZ) were evaluated in 42 patients with respiratory tract infections, including pneumonia (29 patients) and lower respiratory tract infections (5 patients) . Overall clinical efficacy rates (excellent + good) were 79% in pneumonia and 80% in respiratory tract infections in 34 patients evaluated for clinical efficacy . It was excellent that the clinical efficacy rate was 92% in mild and moderate pneumonia . Pathogens isolated from sputa were 31 strains, including 8 strains of Pseudomonas aeruginosa, 7 of Streptococcus pneumoniae, 3 of Staphylococcus aureus and 3 of Haemophilus influenzae . Since the isolates were eradicated in 18 strains, replaced in 3, unchanged in 2 and unknown in 8, the overall eradication rate was 91% . The eradication rates were 89% in beta-lactamase producing strains and 100% in beta-lactamase positive sputum, and excellent or good in 19 (83%) of 23 patients with beta-lactamase negative sputum . The eradication rate was 88% in 5 patients with beta-lactamase positive sputum . One patient experienced a moderate rash . Abnormal laboratory test values were observed in 10 patients (26.3%), but these abnormalities were mild and transient . These results suggested that SBT/CPZ was effective and safe for the treatment of respiratory tract infections caused by beta-lactamase producing as well as beta-lactamase non-producing bacteria.

Arch Pediatr, 1996 Aug, 3(8), 775 - 81
{Trial of prevention of severe Haemophilus influenzae type b infections and trial of tolerance, after PRP-T vaccination, in the Val-de-Marne region}; Boucher J et al.; BACKGROUND: Protein-polysaccharide conjugate H influenzae vaccine is now routinely recommended for infants . To assess the vaccine's protective efficacy against invasive H influenzae infections and its safety, we conducted a study in the Val-de-Marne area of France . METHODS: From April 1991 to April 1993, 22,443 children less than 5 years of age were given PRP-T vaccine . Infants less than 6 months were given three doses whereas those between 6 and 12 months received only two doses, and children over 1 year of age received one dose . According to the infant's DTP-IPV vaccination status, PRP-T was administered alone or reconstituted extemporaneously with DTP-IPV . The immunogenicity of the conjugate vaccine was assessed after three doses in 100 infants under the age of 6 months . RESULTS: The PRP-T vaccine administered alone was safe . The reactions were more frequent when PRP-T vaccine was combined with DTP-IPV vaccine but they were comparable in frequency and severity to those observed after DTP-IPV vaccination . Before 1992, 18 Hib infections were reported each year in the Val-de-Marne region . During the study, only three Hib infections were reported each year . CONCLUSIONS: The fall in incidence of Hib infections, greater than expected, suggests a widespread immune effect of the vaccine, possibly due to a decrease in Hib nasopharyngeal carriage . The antibody titres to each component of vaccine were comparable to those observed in previous clinical infant studies.

Singapore Med J, 1996 Aug, 37(4), 374 - 7
Severe community-acquired pneumonia in Singapore; Lee KH et al.; BACKGROUND: There has been no previous study documenting the aetiology and prognosis of severe community-acquired pneumonia in Singapore . Patients with severe community-acquired pneumonia (SCAP) needing admission to a medical intensive care unit in Singapore were studied retrospectively . METHODS: All patients admitted to the medical intensive care unit at National University Hospital from June 1991 to February 1993 with a diagnosis of community-acquired pneumonia were entered into the study . All patients had blood cultures, sputum cultures, serologies for Legionella and mycoplasma drawn on admission . APACHE II scores were determined prospectively . RESULTS: Fifty-nine consecutive cases from June 1991 to February 1993 were identified with a mean age of 61 (SD 17) years . Nearly all the cases needed mechanical ventilation (90%) and overall mortality was 63% . An aetiological agent was identified in the majority of cases (68%), with Klebsiella pneumoniae being the most common agent (9 cases, 15%) . Haemophilus influenzae and Streptococcus pneumoniae were identified in 8% (5 cases) and 5% (3 cases) of cases respectively . Pseudomonas pseudomallei was identified in 4 cases (7%) with a 100% mortality . Overall, gram-negative organisms were identified in 47% of cases . APACHE II was significantly higher in non-survivors . Age, creatinine levels, and the presence of bacteraemia were not prognostic features . CONCLUSION: SCAP in Singapore carries a high mortality with the predominance of gram-negative organisms . Empiric antibiotics should include gram-positive and gram-negative coverage with specific coverage for Pseudomonas pseudomallei.

Diagn Microbiol Infect Dis, 1996 Aug, 25(4), 213 - 7
Antimicrobial resistance issues of the future; Craig WA; Increasing antimicrobial resistance among respiratory pathogens has the potential to reduce the efficacy of standard dosage regimens for many oral drugs . The goal of antimicrobial therapy is to maximize bactericidal activity . The duration of time that serum concentrations exceed the MIC is the pharmacokinetic/pharmacodynamic parameter that determines efficacy for beta-lactams, macrolides, and trimethoprim/sulfamethoxazole . Studies in animal models suggest that serum levels of beta-lactams need to exceed the MIC for about half of the dosing interval to obtain maximum antimicrobial efficacy . Studies in children with acute otitis media also demonstrate that serum concentrations need to exceed the MIC for 40% or more of the dosing interval to obtain bacteriologic cure in over 85% of patients . With the oral beta-lactams used against penicillin-resistant Streptotoccus pneumoniae, this goal is obtained only with amoxicillin and amoxicillin/clavulanate . For Haemophilus influenzae, several beta-lactams including cefixime, cefpodoxime, and amoxicillin/clavulanate provide serum levels with the longest durations above the MIC . Antimicrobial resistance has also stimulated the search for new potent antimicrobials, altered but effective dosing regimens, and resistance control measures, such as the prudent use, optimal infection control practices, and vaccines to reduce colonization and subsequent infection.

Diagn Microbiol Infect Dis, 1996 Aug, 25(4), 183 - 90
A multicenter study of the antimicrobial susceptibility of community-acquired lower respiratory tract pathogens in the United States, 1992-1994 . The Alexander Project; Washington JA; A multicenter, collaborative study was performed over a three-year period (1992-1994) to determine the antimicrobial susceptibilities of isolates of Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae from community-acquired lower respiratory tract infections . Isolates were collected from five geographically separated medical centers in the United States and sent to a central laboratory for antimicrobial susceptibility testing . Of 350, 536, and 372 isolates of H . influenzae collected from the five centers in 1992, 1993, and 1994, 26.3%, 28.2%, and 30.1%, respectively, were beta-lactamase-positive . All isolates of H . influenzae remained susceptible to ceftriaxone, doxycycline, ciprofloxacin, and ofloxacin over the three-year period . Between 95 and 100% of isolates of H . influenzae remained susceptible to amoxicillin-clavulanic acid cefixime, clarithromycin, and chloramphenicol over this same period . The prevalence of beta-lactamase-positive isolates of M . catarrhalis increased from 92.1% in 1992 to 93.8% in 1993 and to 96.5% in 1994; however, isolates of this species were highly susceptible to amoxicillin-clavulanic acid, the cephalosporins, the macrolides, the fluoroquinolones, chloramphenicol, doxycycline, and trimethoprim-sulfamethoxazole . The prevalence of penicillin-intermediate isolates of S . pneumoniae decreased from 16% in 1992 to 8.2% in 1994, whereas that of penicillin-resistant isolates increased from 5.6% to 10.9% in the same period . Ceftriaxone susceptibility declined from 95.2% to 88.4% over the three years, whereas chloramphenicol susceptibility declined from 98.4% to 90.5% and that of trimethoprim-sulfamethoxazole declined from 85.6% to 68.7% . Macrolide activity remained unchanged.

Diagn Microbiol Infect Dis, 1996 Aug, 25(4), 169 - 81
Results of the Alexander Project: a continuing, multicenter study of the antimicrobial susceptibility of community-acquired lower respiratory tract bacterial pathogens; Gruneberg RN et al.; In 1992, an ongoing, international multicenter study was established to investigate the antimicrobial susceptibility of community-acquired lower respiratory tract bacterial pathogens: the Alexander Project . Isolates cultured from patients living in geographically separated areas, ten in the European Union (EU) and five in the United States (US), were collected and tested using standard methods in a central laboratory . A total of 4,155 isolates of Haemophilus influenzae was collected during the period 1992-1994 . beta-lactamase production was the principal mechanism of resistance observed with overall rates in the US (1992 = 26.3%; 1993 = 28.2%; and 1994 = 30.1%) generally twice those seen in the EU (1992 = 12.3%; 1993 = 14.4%; and 1994 = 15.5%) . Chloramphenicol resistance was generally low except in Spanish centers where rates ranging from 4.0 to 15.9% were observed during the study period . One thousand one hundred ninety-three isolates of Moraxella catarrhalis were tested . beta-lactamase production was the only mechanism of resistance of any importance detected, with the vast majority of isolates producing the enzyme . Two thousand eight hundred twenty-nine isolates of Streptococcus pneumoniae were tested . French and Spanish centers provided isolates with the highest rates of either low-level (intermediate) or high-level penicillin resistance, which in 1994 ranged from 10.2 to 31.4% and 30.4 to 40.1% for each resistance category, respectively . With the exception of the fluoroquinolones, rates of resistance to other antimicrobials including the macrolides, doxycycline, chloramphenicol, and trimethoprim/sulfamethoxazole were high, generally, in centers with a high prevalence of penicillin resistance . However, in some centers (Toulouse, France and Genoa, Italy) this association was not complete for the macrolides.

J Clin Pharm Ther, 1996 Aug, 21(4), 255 - 60
Guidelines for antimicrobial prophylaxis; Nahata MC; Antimicrobials are frequently used to prevent infections . Principles of prophylaxis, and antimicrobial prophylaxis in surgery, tuberculosis, acquired immunodeficiency syndrome, influenza A, traveller's diarrhoea, malaria, recurrent otitis media, Haemophilus influenzae type b infection, pertussis, rheumatic fever, and urinary tract infection are described . Various strategies to improve the prophylactic use of antibiotics are discussed . Collaborative efforts among health care disciplines are needed to assure optimal antimicrobial prophylaxis . This should maximize efficacy and minimize adverse effects, the development of bacterial resistance and associated costs.

East Afr Med J, 1996 Aug, 73(8), 527 - 32
Post-endemic acute bacterial meningitis in Sudanese children; Ahmed AA et al.; The study describes an 18-month surveillance of the epidemiological, clinical and microbiological features of childhood acute bacterial meningitis (ABM) seven months after the end of a major epidemic of Neisseria meningitidis (MC) in Khartoum . A total of 125 children, aged one month to 14 years, who were admitted with a provisional diagnosis of meningitis/meningoencephalitis to the Children's Emergency Hospital (CEH) in Khartoum, Sudan, were prospectively enrolled in the study . Bacterial meningitis was diagnosed by direct microscopy (DM), culture or a recently introduced ELISA assay (EIA-test) in 56 children . Haemophilus influenzae (HI) and MC were the commonest causative bacteria (each accounting for 38%) and were followed by Streptococcus pneumoniae (PNC, 23%) . There was a relative decrease in the proportion of HI which was found to be the leading causative bacteria in a previous study done in the same hospital during endemic situations . This was accounted for by a relative excess of MC during the post-epidemic period . Molecular analysis of two MC strains revealed that clone III-I of serogroup A (that caused an intercontinental wave of MC disease between 1983 and 1990) was still prevalent . The case fatality rate was 28.6% which is higher than that reported in Sudan (18.6%) during endemic situations; but comparable to the mortality in other African countries . Of the prognostic factors on admission, low systolic blood pressure (< 70 mmHg), hyperpyrexia (temperature > 40 degrees C) and light to deep coma correlated significantly with a fatal outcome.

Int J STD AIDS, 1996 Aug-Sep, 7(5), 324 - 32
STD care in Zambia: an evaluation of the guidelines for case management through a syndromic approach; Hanson S et al.; Clinical diagnosis of STDs is unreliable and therefore constitutes a poor basis for choice of treatment . A syndromic approach has been suggested to increase effectiveness of treatment in resource poor settings . Algorithms for the treatment of STD syndromes were evaluated . A total of 436 patients were followed; cure rates were defined and estimated for genital ulcer disease (GUD), urethral and vaginal discharge . Cure rates for the discharge syndromes were high, 97-98%, for both sexes . The cure rate for GUD was 83% for female and 69% for male patients . A higher prevalence of syphilis in the female study population probably contributed to this . It is likely that a large proportion of the treatment failures were due to decreased susceptibility of Haemophilus ducreyi to trimethoprim-sulpha . The determination of cure rates met with a number of methodological problems . This makes it difficult to evaluate the algorithms as part of routine activities, as suggested earlier by WHO.

Kansenshogaku Zasshi, 1996 Aug, 70(8), 808 - 14
{A clinical features of acute respiratory infection with Haemophilus influenzae by transtracheal aspiration}; Tsujimoto M et al.; We performed a clinical study of 26 cases (27 episodes) of acute respiratory infection with H . influenzae by trans tracheal aspiration (TTA) from May 1987 to April 1995 . 15 episodes (14 cases) were bronchitis and 12 episodes (12 cases) were pneumonia . 8 episodes were monomicrobial infection and 19 episodes were polymicrobial infection . Compared to the group of patients of monomicrobial infection, the number of elderly patients and the levels of WBC and CRP were higher in the group of patients of monomicroibal infection . In bronchitis cases, monomicrobial infections of H . influenzae were 7 episodes and polymicrobial infection containing H . influenzae were 8 episodes . In the latter group, PaO2 level was lower and CRP was higher on average . All patients recovered, but the period for treatment was longer in the latter group . In the pneumonia group, only one episode was monomicrobal infection and 11 episodes were polymicrobial infection . Inspite of treatment, one patient died . It was considered that polymicrobial infection was an important factor of acute respiratory infection with H . influenzae.

Kansenshogaku Zasshi, 1996 Aug, 70(8), 784 - 91
{Study on septicaemia in infants and children in the past 20 years . Part 2 . An analysis of factors that prescribe for the prognosis}; Sato T et al.; Underlying diseases, complications, clinical findings, and laboratory findings were evaluated in 158 cases of septicaemia admitted to Jikei University Hospital from 1975 to 1994, in order to conjectured factors that prescribe for the prognosis . 50% of the patients had underlying diseases . Malignancy including leukaemia (31 cases, 39.2%) was the most common underlying disease, followed by low birth weight infant (17 cases, 21.5%), aplastic anemia (9 case, 11.4%), and congenital heart disease (7 cases, 8.9%) . The death rate for patients with underlying disease (27.8%) was significantly greater than the mortality for normal patients with septicaemia (8.9%) (p < 0.05) . Meningitis (24.7%) was the most common complication, followed by DIC (19.6%), shock (15.2%), and pneumonia (10.8%) . The mortality rate of septicaemia complicated by shock was 66.7% (p < 0.01), and that complicated by DIC was 45.2% (p < 0.01) . The mortality rate for patients with the clinical findings of respiratory distress, cough, abdominal distention, cyanosis, splenomegaly, or peripheral coldness was more than 40% and significantly greater (p < 0.01) . Mortality rate in patients with granulocyte counts of < 4.000/mm3, platelet counts of < 5 x 10(4)/ mm3, total protein of < 5.0 g/dl, or ESR of < 20 mm/hr were significantly greater (p < 0.01) than those in patients with normal laboratory findings . Coincidence rate of blood and stool cultures was 57.9% for E . coli, and 28.6% for Klebsiella sp., and that of blood and throat cultures was more than 30% for Pseudomonas sp., Haemophilus influenzae, and Staphylococcus aureus . In the study of antimicrobial susceptibility for microorganisms isolated, the number of drug resistant S . aureus had increased in the last 10 years.

J Formos Med Assoc, 1996 Aug, 95(8), 599 - 604
Invasive Haemophilus influenzae diseases and purulent meningitis in Taiwan; Wang CH et al.; We conducted a 3-year Taiwan-wide hospital-based survey of invasive Haemophilus influenzae infections in children less than 15 years of age . From January 1992 to December 1994, 105 cases (57 boys, 48 girls) were reported . Seventy-three patients (69.5%) had meningitis and 32 patients had other diseases (12 pneumonia, 10 sepsis, 7 cellulitis, 3 arthritis) . Fourteen patients (13%) died, all of whom had meningitis or sepsis . Among the 63 patients who survived meningitis, 17 (27%) had neurologic sequelae and eight (47%) had hearing impairment . The number of cases of H . influenzae meningitis (30%) and other H . influenzae diseases (29%) peaked in children between 6 and 12 months of age . Patients with invasive infections (82%) and meningitis (73%) were younger than 24 months of age . Only 12 patients (11%) were older than 5 years of age and four had underlying diseases . The annual incidence of invasive H . influenzae infections in children less than 5 years old was 1.9 per 100,000 per year . During the same period a survey of purulent meningitis in children younger than 15 years of age was also conducted in 20 hospitals . A total of 198 patients, in whom the causative organisms were identified, were included; 94 patients were 2 months of age or under and the most frequent pathogen was group B streptococci (35 cases, 37%) . Among the 104 patients who were older than 2 months of age, H . influenzae was the leading cause (38 cases, 37%) . In conclusion, invasive H . influenzae type b (Hib) diseases exist in Taiwan but have an incidence lower than in Western countries . Hib meningitis is still the most common cause of purulent meningitis in children in Taiwan and is an important cause of mortality and morbidity . Continuous active surveillance of invasive H . influenzae infections is suggested to determine the best time to introduce an Hib conjugate vaccine in Taiwan.

Arch Dis Child, 1996 Aug, 75(2), 159 - 61
Antibody responses to Haemophilus influenzae type b conjugate vaccine in sickle cell disease; Goldblatt D et al.; OBJECTIVE: To investigate the immunogenicity of Haemophilus influenzae type b (Hib) conjugate vaccines in children with sickle cell disease . DESIGN: Open study . SETTING: Haemoglobinopathy clinic . SUBJECTS: Children with homozygous haemoglobin SS disease (HbSS), sickle haemoglobin C disease (HbSC), and sickle-beta thalassaemia disease (HbS-beta Thal) . INTERVENTIONS: Children over the age of 2 years received a single dose of Hib-tetanus toxoid conjugate vaccine (PRP-T) . MAIN OUTCOME MEASURES: Antibody response to Hib polysaccharide (PRP) approximately one month after vaccination . RESULTS: 77 children over the age of 2 years were studied,, 55 with HbSS, 16 with HbSC, and six with HbS-beta Thal . Before vaccination, 44% had anti-PRP IgG titres less than the level associated with long term protection (1.0 microgram/ml) . After a single dose of PRP-T all children mounted an antibody titre > 1 microgram/ml . Geometric mean anti-PRP IgG titre achieved postvaccination (45.2 micrograms/ml 95% confidence interval (CI) 31.6 to 64.8) was comparable to that of a healthy population . Children with HbSC, however, had a significantly higher antibody titre postvaccination (91.1 micrograms/ml; 95% CI 32.7 to 254.4) than the children with HbSS (36.7 micrograms/ml; 95% CI 25.1 to 52.9) . CONCLUSIONS: Children with a diagnosis of sickle cell disease who are over the age of 2 years make a vigorous antibody response to a single dose of PRP-T vaccine and hence we suggest unimmunised individuals in this group should receive a single dose of a Hib conjugate vaccine.

J Otolaryngol, 1996 Aug, 25(4), 249 - 56
Microbiology and management of sinusitis; Brook I; PURPOSE: The pathophysiology, microbiology, and treatment of acute and chronic sinusitis are reviewed . MATERIALS AND METHODS: The current literature is reviewed . REVIEW: Sinusitis generally develops as a complication of viral or allergic inflammation of the upper respiratory tract . Bacterial pathogens contribute to the inflammatory process-Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis are predominant in acute sinusitis, while anaerobic bacteria and Staphylococcus aureus are predominant in chronic sinusitis . Pseudomonas aeruginosa has emerged as a potential pathogen in the immunocompromised patients and in those who have nasal tubes or catheters . Many of these organisms recovered from sinusitis became resistant to penicillins either through the production of beta-lactamase (H . influenzae, M . catarrhalis, S . aureus, Fusobacterium sp., and Prevotella sp.) or through changes in the penicillin-binding protein (S . pneumoniae) . The pathogenicity of beta-lactamase-producing bacteria is expressed directly through their ability to cause infections, and indirectly through the production of beta-lactamase . CONCLUSIONS: The indirect pathogenicity is conveyed not only by surviving penicillin therapy, but also by "shielding" penicillin-susceptible pathogens from the drug . The direct and indirect virulent characteristics of these bacteria require the administration of appropriate antimicrobial therapy directed against all pathogens in mixed infections . Proper therapy, with antimicrobial and decongestants combined when indicated with surgical drainage and correction of pathology, constitutes the cornerstone of management of sinusitis.

Protein Sci, 1996 Aug, 5(8), 1704 - 18
Topology prediction for helical transmembrane proteins at 86% accuracy; Rost B et al.; Previously, we introduced a neural network system predicting locations of transmembrane helices (HTMs) based on evolutionary profiles (PHDhtm, Rost B, Casadio R, Fariselli P, Sander C, 1995, Protein Sci 4:521-533) . Here, we describe an improvement and an extension of that system . The improvement is achieved by a dynamic programming-like algorithm that optimizes helices compatible with the neural network output . The extension is the prediction of topology (orientation of first loop region with respect to membrane) by applying to the refined prediction the observation that positively charged residues are more abundant in extra-cytoplasmic regions . Furthermore, we introduce a method to reduce the number of false positives, i.e., proteins falsely predicted with membrane helices . The evaluation of prediction accuracy is based on a cross-validation and a double-blind test set (in total 131 proteins) . The final method appears to be more accurate than other methods published: (1) For almost 89% (+/-3%) of the test proteins, all HTMs are predicted correctly . (2) For more than 86% (+/-3%) of the proteins, topology is predicted correctly . (3) We define reliability indices that correlate with prediction accuracy: for one half of the proteins, segment accuracy raises to 98%; and for two-thirds, accuracy of topology prediction is 95% . (4) The rate of proteins for which HTMs are predicted falsely is below 2% (+/-1%) . Finally, the method is applied to 1,616 sequences of Haemophilus influenzae . We predict 19% of the genome sequences to contain one or more HTMs . This appears to be lower than what we predicted previously for the yeast VIII chromosome (about 25%).

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