Haemophilus

Diagn Microbiol Infect Dis, 1997 Aug, 28(4), 205 - 8
Vertebral osteomyelitis: nontypeable beta-lactamase-negative Haemophilus influenzae in an adult: case report; Personius CD et al.; Adult pyogenic vertebral osteomyelitis due to Haemophilus influenzae is exceedingly rare . After a search of the literature, we deemed our case to be the seventh case of H . influenzae pyogenic osteomyelitis . Vertebral osteomyelitis in itself is a rarity . The most common organisms associated with vertebral osteomyelitis are Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa . Of the six previously reported cases of adult pyogenic vertebral osteomyelitis due to H . influenzae, four of the six cases were caused by Type B H . influenzae, one case was attributed to Type C, and the other strain was not typed.

Arch Microbiol, 1997 Nov, 168(5), 345 - 54
Molybdate transport and regulation in bacteria; Grunden AM et al.; Molybdate is transported in bacteria by a high-affinity transport system composed of a periplasmic binding protein, an integral membrane protein, and an energizer protein . These three proteins are coded by modA, modB, and modC genes, respectively . The ModA, ModB, and ModC proteins from various organisms (Escherichia coli, Haemophilus influenzae, Azotobacter vinelandii, and Rhodobacter capsulatus) are very similar . The lowest Km value reported for molybdate in the molybdate transport process is approximately 50 nM . In a mod mutant, molybdate is transported by the sulfate transport system or by a nonspecific anion transporter . Molybdate transport is tightly coupled to utilization in E . coli and Klebsiella pneumoniae, while other dinitrogen-fixing organisms appear to have a molybdenum storage protein . In all organisms studied so far, molybdate transport genes are regulated by a repressor protein, ModE . The ModE-molybdate complex binds to the sequences TAYAT (Y = T or C) in the operator/ promoter region in E . coli and prevents transcription of the modABCD operon . The ModE-molybdate complex binds to DNA as a homodimer in E . coli and possibly in other organisms as well . In R . capsulatus, however, two ModE homologues (MopAB proteins) are required for repression.

Pharmacotherapy, 1997 Sep-Oct, 17(5), 985 - 9
Microbiologic effect of bovine cerebrospinal fluid and azithromycin against Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae; Destache CJ et al.; Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of azithromycin against reference strains of Streptococcus pneumoniae ATCC 49619, Neisseria meningitidis ATCC 13090, and Haemophilus influenzae ATCC 49247 were determined by the macrodilution broth method with and without 10% bovine cerebrospinal fluid (CSF) supplementation . The MICs and MBCs were within one to two dilutions for N . meningitidis and S . pneumoniae, and no difference was observed for H . influenzae . Time-kill curves demonstrated enhanced killing by azithromycin when 10% bovine CSF was added to media for N . meningitidis . The minimum azithromycin concentration for a greater than 3 log10 reduction in inoculum with bovine CSF was 0.03 microg/ml and without CSF was 0.12 microg/ml, a 3-fold difference . Killing was not significantly different for either H . influenzae nor S . pneumoniae.

Zentralbl Veterinarmed B, 1997 Sep, 44(7), 405 - 13
Association of tumour necrosis factor and acute phase reactant changes with post arrival disease in swine; Harding JC et al.; Severe weight loss in the absence of respiratory, enteric or systemic clinical disease or gross pathologic lesions is often observed when immunologically naive boars are placed in conventional health swine facilities . Affected animals develop this weight loss in spite of receiving pre-entry vaccinations against common swine pathogens, such as Haemophilus parasuis or Mycoplasma hyopneumoniae . In many cases, the weight loss is non-responsive to long term antibiotic therapy . In order to determine the relationships between the severity of post arrival weight loss and disease and its potential immunological or physiological indicators, tumour necrosis factor (TNF) and acute phase reactant levels were correlated with the clinical status in immunologically naive boars following their transfer to a conventional facility . Boars had higher TNF (P < 0.0001) and plasma protein (P = 0.0054) levels and decreased zinc (P = 0.0004) levels during periods of clinical sickness . Likewise, peak and average plasma TNF, serum haptoglobin, and serum zinc were correlated indicating a prolonged stress or pathogenic insult (r = 0.89, P < 0.0001 for TNF; r = 0.67, P = 0.01 for haptoglobin; r = 0.73, P = 0.005 for zinc) . An acute phase response, a systemic TNF increase and the development of a lymphopenia were observed in post arrival disease in swine . This is the first time cytokines and acute phase reactants have been investigated in a field study involving immunologically naive or high health swine.

J Paediatr Child Health, 1997 Aug, 33(4), 296 - 9
Meningitis in Hong Kong children, with special reference to the infrequency of haemophilus and meningococcal infection; Sung RY et al.; OBJECTIVE: To study the epidemiologic and aetiologic features of meningitis in children in Hong Kong . METHODOLOGY: A retrospective study of 85 children resident in the New Territory East region of Hong Kong admitted to a teaching Hospital because of meningitis during a 9 year period . RESULTS: Mycobacterium tuberculosis was the most common aetiological agent accounting for 13 cases (15.3%) . Other bacteria accounted for 41 cases (48%); among these one fifth were caused by Haemophilus influenzae type b . The overall admission rates for tuberculous meningitis in Chinese children were 0.76/100,000 (95% CI 0.25-1.78) and 0.42/100,000 (CI 0.19-0.8) per year, respectively, for under 5 year olds and under 15 year olds . The overall annual incidence rates of bacterial meningitis other than tuberculous were 5.2/100,000 (CI 3.72-7.43) and 1.6/100,000 (CI 1.14-2.29) for Chinese children under 5 years and under 15 years, respectively . The annual incidence of H . influenzae meningitis in Chinese children under 5 years old was low at 1.1/100,000 (0.43-2.2) . All five cases of meningococcal meningitis were in Vietnamese children (under 5 years of age incidence: 13.0/100,000 per year, CI 4.2-30.3) . There were no cases of meningococcal meningitis in Chinese children during the 9 year period . CONCLUSION: M . tuberculosis was the most common aetiological agent of meningitis in Hong Kong children . The incidence of haemophilus or meningococcal meningitis was very low.

Pediatr Infect Dis J, 1997 Sep, 16(9), 863 - 70
Vaccine antigen interactions after a combination diphtheria-tetanus toxoid-acellular pertussis/purified capsular polysaccharide of Haemophilus influenzae type b-tetanus toxoid vaccine in two-, four- and six-month-old infants; Pichichero ME et al.; OBJECTIVE: The safety and immunogenicity of a diphtheria-tetanus toxoid-acellular pertussis vaccine (DTaP; Trepedia)/Haemophilus influenzae b polysaccharide (PRP-T; ActHib) combined vaccine (TriHibir; Pasteur Merieux Connaught) was compared with DTaP and PRP-T given at the same visit but at separate sites in a prospective multicenter, open label trial . METHODS: Infants were randomized to four groups (three consistency lots of DTaP/PRP-T vs . one of the consistency lots given as separate vaccines); injections were administered at 2, 4 and 6 months of age . Pre-Dose 1 and post-Dose 3 sera were assayed for antibody titers against all antigens . Reactions to the vaccinations were assessed by parent questionnaire for 30 days after each injection visit . RESULTS: Four hundred eighty-five infants were enrolled; 296 evaluable infants were included in the DTaP/PRP-T group compared with 70 infants in the DTaP and PRP-T vaccine group . Infants who received the combined vaccine had higher post-Dose 3 geometric mean antibody titers to diphtheria antitoxin (P < 0.01) and pertussis filamentous hemagglutinin (P < 0.05) and lower geometric mean antibody titers to tetanus antitoxin (P < 0.05) and Haemophilus influenzae b (Hib) polysaccharide (PRP) (P < 0.05) . The geometric mean anti-PRP antibody titer in the DTaP/PRP-T group was 4.3 micrograms/ml compared with 7.0 micrograms/ml in the separate vaccine group (P < 0.05), and the percentage of infants with antibody titers > or = 0.15 and 1 microgram/ml were, respectively, 95 and 86%, whereas they were 100% for both titers in the separate vaccines group . DTaP/ PRP-T vaccine given concomitantly or 1 month apart from hepatitis B vaccine and oral poliomyelitis vaccine caused no significant differences in immunogenicity or safety . The safety assessments for the DTaP/PRP-T vaccine showed no consistent differences in systemic or local injection site reactions compared with DTaP and PRP-T administered separately . CONCLUSION: Although the antibody responses to tetanus and Hib polysaccharide in the evaluated DTaP/PRP-T combined vaccine were significantly lower than those seen after separate DTaP and PRP-T administration, the combined vaccine elicited an immune response against diphtheria, tetanus, pertussis and Haemophilus influenzae b likely to confer protection.

Pediatr Infect Dis J, 1997 Sep, 16(9), 858 - 62
Chlamydia pneumoniae in acute otitis media; Block SL et al.; BACKGROUND: Aerobic bacterial pathogens are recovered from 65 to 85% of patients with acute otitis media (AOM) . Although Chlamydia pneumoniae is a common pathogen of pediatric pneumonia, it has rarely been cultured from children with chronic otitis media and its role in AOM is unknown . METHODS: We cultured for C . pneumoniae in tympanocentesis aspirates and nasopharyngeal swabs from 101 consecutive, otherwise healthy children with AOM or refractory AOM . A control group of 50 similarly aged, healthy children was evaluated for nasopharyngeal carriage of C . pneumoniae . Specimens were also evaluated by PCR for C . pneumoniae . RESULTS: C . pneumoniae was recovered by tympanocentesis in 8 (8%) of 101 children with AOM . Among the 8 children with C . pneumoniae-positive-AOM, 5 had C . pneumoniae detected by PCR in middle ear fluid, none had C . pneumoniae recovered by nasopharyngeal culture or PCR and 5 were younger than 16 months . C . pneumoniae was the sole pathogen isolated in 2 patients . Copathogens included beta-lactamase-positive positive Haemophilus influenzae (2), beta-lactamase positive Moraxella catarrhalis (1), penicillin-resistant Streptococcus pneumoniae (2) and penicillin-susceptible S . pneumoniae (1) . C . pneumoniae was recovered from nasopharyngeal culture in 2 additional patients with C . pneumoniae-negative AOM and in none of 50 healthy control children, although 2 controls were positive by PCR from the nasopharynx . CONCLUSIONS: This is the first study to report the isolation of C . pneumoniae in middle ear fluid of children with AOM.

Pediatr Infect Dis J, 1997 Sep, 16(9), 852 - 8
Nasopharyngeal colonization in Costa Rican children during the first year of life; Vives M et al.; BACKGROUND: The establishment of the nasopharyngeal flora was followed in Costa Rican children from birth to 1 year of age . METHODS: Nasopharyngeal cultures were obtained at 1 (n = 413), 3 (n = 393), 6 (n = 376) and 12 months (n = 356) of age from children representative of the population in the Puriscal district . Weekly cultures were obtained from a subcohort of these children (n = 101) . Mother-infant diads (n = 95) and preschool children (n = 208) attending day-care centers were also studied . RESULTS: The estimated proportion of colonized children in the population differed markedly depending on the frequency of culture . Quarterly cultures showed a slow increase in carrier rates from 3.9% for Haemophilus influenzae, 3.1% for Streptococcus pneumoniae and 6.5% for Moraxella catarrhalis at 1 month of age to 10.1% carrying H . influenzae and 19.4% carrying S . pneumoniae by the end of the first year . By quarterly culture the proportion of children colonized at least once was 36% for S . pneumoniae, 26% for H . influenzae and 28% for M . catarrhalis . In contrast weekly sampling showed that 95 to 100% of the children were colonized at least once during the first year of life with H . influenzae, S . pneumoniae or M . catarrhalis . Nasopharyngeal carriage of H . influenzae, S . pneumoniae and M . catarrhalis was low in the mothers, and very few mother-infant pairs carried identical bacteria at the same time . In contrast carrier rates were high in the siblings attending day care (H . influenzae 27.9%, S . pneumoniae 39.4%, both organisms 26.6%) . Infants with siblings had significantly higher bacterial carriage at all ages than infants without siblings . CONCLUSIONS: Quarterly nasopharyngeal cultures showed that Costa Rican infants acquire their nasopharyngeal flora at a rate comparable with that for infants in developed countries and that siblings are an important source of the bacteria . Weekly samplings showed that virtually all children were colonized at least once during the first year of life.

Int J Immunopharmacol, 1997 Mar, 19(3), 181 - 6
Kinetics of specific salivary IgA responses in man after oral challenge by ribosomal immunostimulant; Kolopp-Sarda MN et al.; The kinetics of specific IgA mucosal responses was assessed in 12 healthy volunteers over 3 weeks of treatment by oral administration of an immunostimulant, Ribomunyl, composed of ribosomes from the four bacteria Streptococcus pyogenes, Streptococcus pneumoniae, Klebsiella pneumoniae and Haemophilus influenzae . The levels of IgA specific for these four bacteria increased after each immunization and, after the third week of treatment, were significantly higher than baseline day 0 values . This study demonstrates that oral ribosomal immunostimulation results in the production of specific salivary antibodies liable to recognize whole bacteria antigens, and therefore likely to confer protection . The kinetic analysis performed also demonstrates the rapidity of specific mucosal immune responses after oral stimulation in man, a feature still seldom explored.

Indian J Biochem Biophys, 1997 Feb-Apr, 34(1-2), 124 - 30
The minimal cellular genome of mycoplasma; Razin S; The mycoplasmas are the smallest and simplest self-replicating organisms, being built of a plasma membrane, ribosomes, and a circular double-stranded DNA molecule-the typical prokaryotic genome . The idea of using mycoplasmas as models for defining in molecular terms the entire machinery of a living cell was raised by Morowitz in 1984 . The goal has been to prove the dogma of the completeness of molecular biology, that is, that the logic of life is finite, relatively simple and subject to full exploration . The recent complete sequencing of the genome of the human pathogen Mycoplasma genitalium brings us much closer to achieving this goal . The M . genitalium genome is only 580 kb long and contains only 470 predicted coding sequences(genes), as compared with 1727 in Haemophilus influenzae and about 4000 in E . coli . Thus, M . genitalium is apparently the simplest organism capable of independent life with a minimal set of genes . The drastic economization in genetic information must be associated with the parasitic mode of life of the mycoplasmas . Mycoplasmas evolved by reductive evolution from Gram-positive bacteria with low G + C genomes . During evolution the mycoplasmas have lost the cell wall and many biosynthetic systems involved in synthesis of macromolecule building blocks provided by their host . Thus, the M . genitalium genome carries only one gene involved in amino acid biosynthesis, and very few genes for vitamin and nucleic acid precursors; the lack of genes involved in fatty acid biosynthesis, leads to dependence on exogenous fatty acids, enabling the introduction of controlled variations in membrane acyl chains and the use of mycoplasmas as models in studying membrane fluidity . Moreover, the dependence of mycoplasmas on exogenous cholesterol for growth was exploited to show the role of cholesterol as a buffer of membrane fluidity . The mycoplasma genome carries the minimal set of energy metabolism genes, being content with a restricted supply of ATP needed for their parasitic mode of life . Being limited by a single permeability barrier enabled the saving of a considerable number of transport system genes . Nevertheless, these minimal organisms were shown to carry all the essential genes needed for DNA replication, transcription and translation, but even here gene saving is expressed in a minimal number of rRNA and tRNA genes . A genomic price had been paid to maintain parasitism, so that a significant number of mycoplasmal genes is devoted to adhesins, attachment organelles and variable membrane surface antigens directed towards evasion of the host immune system.

J Mol Evol, 1997 Nov, 45(5), 467 - 72
Conserved gene clusters in bacterial genomes provide further support for the primacy of RNA; Siefert JL et al.; Five complete bacterial genome sequences have been released to the scientific community . These include four (eu)Bacteria, Haemophilus influenzae, Mycoplasma genitalium, M . pneumoniae, and Synechocystis PCC 6803, as well as one Archaeon, Methanococcus jannaschii . Features of organization shared by these genomes are likely to have arisen very early in the history of the bacteria and thus can be expected to provide further insight into the nature of early ancestors . Results of a genome comparison of these five organisms confirm earlier observations that gene order is remarkably unpreserved . There are, nevertheless, at least 16 clusters of two or more genes whose order remains the same among the four (eu)Bacteria and these are presumed to reflect conserved elements of coordinated gene expression that require gene proximity . Eight of these gene orders are essentially conserved in the Archaea as well . Many of these clusters are known to be regulated by RNA-level mechanisms in Escherichia coli, which supports the earlier suggestion that this type of regulation of gene expression may have arisen very early . We conclude that although the last common ancestor may have had a DNA genome, it likely was preceded by progenotes with an RNA genome.

Pediatr Med Chir, 1997 May-Jun, 19(3), 211 - 4
{Subdural effusion during bacterial meningitis}; Calabri G et al.; Twelve children of age ranged from 4 to 34 months with Haemophilus influenzae type b meningitis treated at Meyer Hospital of Florence, were retrospectively reviewed . Eight patients had subdural effusion demonstrated with TC, RM and transfontanellar ultrasonography . All patients are cured without sequelae.

Kansenshogaku Zasshi, 1997 Sep, 71(9), 890 - 4
{An epidemiological investigation for gram-positive coccus, especially PRSP, in Kinki area}; Tsunoda T et al.; An epidemiological investigation for penicillin-resistant Streptococcus pneumonia (PRSP) was performed at 18 medical institutes in Kinki area by the questionnaire from Kinki Infection Working Group 1995 . This investigation was the first report that was performed for a long term (one year) and a large area . The most frequent specimen was sputum from out-patients (50.3%) and inpatients (48.8%), and especially from spinal fluid of 3 cases were detected . Polymicrobial infection with more than 3 pathogens was 15.7%, and it was more frequent than MRSA previously investigated . Simultaneous pathogens detected with PRSP were Candida species, Haemophilus influenzae and Staphylococcus aureus . In terms of chemosusceptibility, VCM (100%), FMOX (97.9%), IPM/CS (85.9%), CEZ (93.4%) and CDTR-PI were determined to be high by sensitive . However, the sensitivity of CCL, which was one of the most common antibiotics, was only 37.7%.

S Afr Med J, 1997 Aug, 87(8 Suppl), 1052 - 4
Bacterial meningitis in children at Kalafong Hospital, 1990-1995; Grobler AC et al.; OBJECTIVES: To determine the bacteriological profile, morbidity, mortality and factors which contributed to the outcome . DESIGN: Retrospective descriptive . SETTING: Kalafong Hospital, Pretoria . SUBJECTS: Children older than 1 month and younger than 13 years admitted between January 1990 and December 1995 with culture-proven bacterial meningitis . MAIN OUTCOME MEASURES: Causative bacteria, antibiotic sensitivity, age, clinical features and short-term morbidity and mortality . RESULTS: Sixty-one patients had culture-proven bacterial meningitis . In 57% Haemophilus influenzae, in 33% Streptococcus pneumoniae and in 1.6% (1 patient) Neisseria meningitidis were cultured (5 patients yielded other organisms) . Some H . influenzae cultures showed resistance to cefotaxime and ampicillin and some S . pneumoniae cultures to penicillin . The mean age of all patients was 18 months . Twenty-three per cent of the patients had consulted between 1 and 5 health workers and were then self-referred . Fever, cough and vomiting were the most common complaints . On examination, fever and pneumonia were the most common findings apart from neurological system involvement . Twelve patients (20%) died and 46% were discharged with obvious neurological complications . Morbidity and mortality were the highest for S . pneumoniae meningitis . Bad prognostic features were delayed referral, S . pneumoniae meningitis, convulsions and abnormal neurological signs on admission . CONCLUSION: H . influenzae was the most common cause of childhood bacterial meningitis at Kalafong Hospital, with N . meningitidis meningitis being very rare . Morbidity and mortality were very high, compared with outcome in developed countries . Delayed referral seems to be an important factor in predicting an adverse outcome.

J Antimicrob Chemother, 1997 Sep, 40(3), 387 - 92
Increasing antimicrobial resistance in Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis in Finland; Manninen R et al.; Respiratory and otitis isolates of 807 Streptococcus pneumoniae, 816 Haemophilus influenzae and 446 Moraxella catarrhalis were collected from 21 clinical microbiology laboratories for antimicrobial susceptibility testing in 1995 . After a period of relative stability in 1981 and 1987-1990, beta-lactamase production increased in H . influenzae . Among middle ear isolates from children under 6 years, beta-lactamase production increased from 8% to 24% in H . influenzae and from 81% to 96% in M . catarrhalis since the survey in 1987-1990 . 1.2% of S . pneumoniae were penicillin-resistant and 4.2% intermediately resistant; 5 years earlier among otitis isolates of children only 1.7% intermediate resistance was found . Ampicillin resistance was seen among 1.9% of non-beta-lactamase-producing strains of H . influenzae . Resistance to trimethoprim-sulphamethoxazole occurred in 9.4% of S . pneumoniae, 7.4% of H . influenzae and 0.7% of M . catarrhalis . Frequencies of azithromycin resistance were 3.0% in S . pneumoniae and 1.6% in H . influenzae, and those of tetracycline resistance were 6.7% in S . pneumoniae and 1.2% in H . influenzae.

Am Fam Physician, 1997 Oct 1, 56(5), 1355 - 62
Issues in the management of bacterial meningitis; Tunkel AR et al.; Acute bacterial meningitis is associated with significant morbidity and mortality despite the availability of effective antimicrobial therapy . The emergence of antibiotic-resistant bacterial strains in recent years has necessitated the development of new strategies for empiric antimicrobial therapy for bacterial meningitis . Specifically, the emergence of strains of Streptococcus pneumoniae that are resistant to penicillin and the cephalosporins have led to empiric therapy for patients with pneumococcal meningitis consisting of vancomycin plus a third-generation cephalosporin pending susceptibility testing . Third-generation cephalosporins are also effective as empiric therapy against other pathogens that cause community-acquired bacterial meningitis, with the exception of Listeria monocytogenes, for which ampicillin or penicillin G is the antimicrobial agent of choice . Adjunctive dexamethasone should be administered to infants and children with suspected or proven Haemophilus influenzae type b meningitis to reduce audiologic and neurologic sequelae; administration concomitant with or just before the first dose of the antimicrobial agent is optimal for best results.

Clin Microbiol Rev, 1997 Oct, 10(4), 585 - 96
Prospects for the development of fungal vaccines; Deepe GS Jr; In an era that emphasizes the term "cost-effective," vaccines are the ideal solution to preventing disease at a relatively low cost to society . Much of the previous emphasis has been on childhood scourges such as measles, mumps, rubella, poliomyelitis, and Haemophilus influenzae type b . The concept of vaccines for fungal diseases has had less impact because of the perceived limited problem . However, fungal diseases have become increasingly appreciated as serious medical problems that require recognition and aggressive management . The escalation in the incidence and prevalence of infection has prompted a renewed interest in vaccine development . Herein, I discuss the most recent developments in the search for vaccines to combat fungal infections . Investigators have discovered several inert substances from various fungi that can mediate protection in animal models . The next challenge will be to find the suitable mode of delivery for these immunogens.

Mol Biol Evol, 1997 Oct, 14(10), 1062 - 74
A comparative study of duplications in bacteria and eukaryotes: the importance of telomeres; Coissac E et al.; The genomes of three bacteria (Haemophilus influenzae, Mycoplasma genitalium, and Escherichia coli) and two eukaryotes (Saccharomyces cerevisiae and Caenorhabditis elegans) were compared . The distribution of their putative open reading frames (ORFs) was studied, and several conclusions were drawn: (1) All of these genomes, even the smallest, exhibit a significant proportion (7%-30%) of duplicated ORFs . This proportion is a function of genome size and appears unrelated to the bacteria/eukaryote division . (2) Some of these ORFs constitute families of up 20 or more members . (3) The levels of sequence similarity within these families are highly variable and their distribution is different among bacteria and eukaryotes . (4) In yeast, there are topological relationships between members of the same family . The paired ORFs are frequently in the same orientation with regard to their respective telomeres and located at comparable distances from them.

Infection, 1997 Sep-Oct, 25(5), 298 - 302
Immunogenicity and reactogenicity of HbOC vaccine administered simultaneously with acellular pertussis vaccine (DTaP) into either arms or thighs of infants; Schmitt HJ et al.; To evaluate the reactogenicity and immunogenicity of a Haemophilus influenzae type b conjugate vaccine (HbOC) and of a tricomponent acellular pertussis vaccine (DTaP) when injected simultaneously into either contralateral arms or into contralateral thighs, 110 infants were enrolled to receive three doses of DTaP at 3, 4, and 5 months and two HbOC doses at 3 and 5 months of age . Administration of either of the two vaccines into arms was associated with significantly more local side effects than administration into thighs . There was no difference in geometric mean concentration (GMC) values for any of the four vaccine antigens between subjects who had been vaccinated into arms or thighs . After immunization, all children had protective antibody titers to diphtheria toxin . While post vaccination the mean anti-tetanus toxoid GMC was > or = 1.25 IU/ml, there was no significant rise as compared to the GMC before vaccination . GMCs of antibodies against the various pertussis antigens were similar to those observed before with the same DTaP vaccine . The simultaneous administration of DTaP and HbOC was safe and immunogenic irrespective of the site of vaccine administration, but significantly more local reactions occurred when vaccines were injected into arms.

Otolaryngol Head Neck Surg, 1997 Sep, 117(3 Pt 1), 263 - 7
Increased secretory capacity of the middle ear mucosa after acute otitis media caused by Haemophilus influenzae type B; Caye-Thomasen P et al.; Secretory otitis media is associated with a highly increased goblet cell density of the middle ear mucosa . Previous studies have shown that a single episode of experimental acute otitis media caused by Streptococcus pneumoniae or nontypeable Haemophilus influenzae is followed by increased goblet cell density for a period of at least 6 months . This condition may create a predisposition for subsequent development of secretory otitis media . We inoculated the middle ears of 25 rats with type B H . influenzae to determine the effect of the bacteria on mucosal secretory capacity . Five rats were euthanized 4, 8, 16, 60, and 180 days after inoculation, followed by dissection, staining, and whole-mount embedding of the middle ear mucosa . The goblet cell density was determined in 24 well-defined localities . Compared with that of 25 normal middle ears, the goblet cell density was significantly increased in almost all counting localities on all days of euthanasia . Thus increased goblet cell density and enlargement of mucosal areas containing goblet cells persisted for 6 months after the acute incident . Inoculation of type B H . influenzae induced an increase of goblet cell density that was higher than the increase after inoculation of S . pneumoniae or nontypeable H . influenzae . We conclude that experimental acute otitis media caused by type B H . influenzae is followed by a longstanding increase of mucosal secretory capacity, which is likely to induce a subsequent development of secretory otitis media.

Antimicrob Agents Chemother, 1997 Oct, 41(10), 2149 - 58
Antibacterial activity of RU 64004 (HMR 3004), a novel ketolide derivative active against respiratory pathogens; Agouridas C et al.; The antibacterial activity of RU 64004, a new ketolide, was evaluated against more than 600 bacterial strains and was compared with those of various macrolides and pristinamycin . RU 64004 had good activity against multiresistant pneumococci, whether they were erythromycin A resistant or not, including penicillin-resistant strains . RU 64004 inhibited 90% of pneumococci resistant to erythromycin A and penicillin G at 0.6 and 0.15 microg/ml, respectively . Unlike macrolides, RU 64004 did not induce the phenotype of resistance to macrolides-lincosamides-streptogramin B . Its good antibacterial activity against multiresistant pneumococci ran in parallel with its well-balanced activity against all bacteria involved in respiratory infections (e.g., Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pyogenes) . In contrast to all comparators (14- and 16-membered-ring macrolides and pristinamycin), RU 64004 displayed high therapeutic activity in animals infected with all major strains, irrespective of the phenotypes of the strains . The results suggest that RU 64004 has potential for use in the treatment of infections caused by respiratory pathogens including multiresistant pneumococci.

Eur Arch Otorhinolaryngol, 1997, 254(8), 372 - 5
Effectiveness of ribosomal fractions of Klebsiella pneumoniae, Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae and the membrane fraction of Kp (Ribomunyl) in the prevention of clinical recurrences of infectious rhinitis . Results of a multicenter double-blind placebo-controlled study; Serrano E et al.; A multicenter, double-blind, placebo-controlled study was conducted to investigate the efficacy of an immunostimulant, Ribomunyl, in the prevention of recurrences of infectious rhinitis in adults . This trial involved 327 patients (168 Ribomunyl treated and 159 placebo cases) with an average of 4.3 +/- 1.8 rhinitis episodes per patient recorded during the year preceding the study . The main criterion of efficacy was the cumulative number of recurrences of infectious rhinitis during a 6-month follow-up period, as analyzed by standard tests . An additional analysis of relative risk of recurrences used multivariate failure for time data . Ribomunyl was effective throughout the study period, starting from the first month of treatment: a mean of 1.0 +/- 1.1 recurrences was recorded in the Ribomunyl group as compared to 1.5 +/- 1.4 recurrences in the placebo group; this indicated one-third fewer infections (P = 0.001) . The protective effect of Ribomunyl on the relative risk for recurrences was estimated to be 0.58 by multivariate analysis (95% CI: 0.43-0.78, P = 0.0001) . Analysis of secondary criteria also favored Ribomunyl: 38.5% less antibiotic courses per patient (0.8 +/- 1.3 vs 1.3 +/- 1.6; P = 0.002) and the number of days with antibiotics (5.6 +/- 9.3 vs 9.1 +/- 12.1; P = 0.002).

Clin Infect Dis, 1997 Aug, 25(2), 206 - 10
Mycobacterium xenopi infection in patients with human immunodeficiency virus infection; el-Helou P et al.; Mycobacterium xenopi is one of the most frequently isolated nontuberculous mycobacteria in Ontario, Canada . We reviewed the records of 28 human immunodeficiency virus (HIV)-infected patients from whom M . xenopi was isolated between 1982 and 1995 . M . xenopi was recovered from respiratory specimens from 24 patients, most of whom had clinical and radiographic evidence of pulmonary disease . However, coexistent pulmonary infection due to other pathogens was found in 17 patients: Pneumocystis carinii (9 patients), cytomegalovirus (5), Haemophilus influenzae (2), Mycobacterium avium complex (2), Streptococcus pneumoniae (1), Staphylococcus aureus (1), Aspergillus species (1), and Histoplasma capsulatum (1) . Three patients had bacteremia with M . xenopi, including two patients with pulmonary infection . Two of the bacteremic patients had chronic fever and a wasting syndrome . Twenty-one (75%) of the 28 patients were thought to be colonized, and seven patients (25%; of whom four had CD4 cell counts of < or = 50/mm3) were thought to have significant infection due to M . xenopi . Sixteen patients died, but in no case was death attributable to M . xenopi infection . In a region where M . xenopi is a relatively common mycobacterial isolate, the organism frequently colonizes HIV-infected patients . Significant disease occurs in those patients with more advanced HIV infection.

Southeast Asian J Trop Med Public Health, 1997 Mar, 28(1), 91 - 8
Safety and immunogenicity of a Haemophilus influenzae type B polysaccharide-tetanus toxoid conjugate vaccine combined with diphtheria, tetanus and pertussis vaccines in Thai infants; Chotpitayasunondh T et al.; A randomized, open, multicenter trial was conducted to determine the safety and immunogenicity of a Haemophilus influenzae type b polysaccharide-tetanus toxoid (PRP-T) conjugate vaccine combined with tetanus, diphtheria and pertussis (DTP) vaccine in 271 Thai infants born to mothers immunized against tetanus during pregnancy . Infants were immunized at approximately 2, 4 and 6 months of age with these vaccines . To determine if elevated levels of anti-tetanus toxin antibodies suppressed the anti-PRP antibody response, a second group of infants were immunized with PRP complexed with outer membrane proteins of Neisseria meningitidis (Pedvax HIB) in one limb at 2 and 4 months of age and DTP vaccine in the other limb at 2, 4 and 6 months of age . A third group of infants received only DTP vaccine at 2, 4 and 6 months of age . The occurrence of both local and systemic adverse reactions were comparable in all 3 groups . The geometric mean anti-tetanus antibody titer was > 1 IU/ml at baseline . Approximately 1 month after the administration of the third dose of vaccine, 98.5%, 99.3% and 9.7% of the children immunized with DTP+Pedvax HIB, DTP-PRP-T or DTP possessed > or = 0.15 microgram of anti-PRP antibody per ml . No child in the DTP group achieved > or = 1 microgram/ml while 74.2% and 89.3% did so after immunization with DTP+Pedvax HIB, or DTP-PRP-T, respectively (p < 0.05) . Immune responses to diphtheria, tetanus and pertussis antigens were similar in all vaccine groups . These results demonstrate that elevated tetanus antibody titers do not diminish the anti-PRP antibody response following immunization with a PRP-T conjugate combined with DTP vaccine.

Gene, 1997 Sep 1, 196(1-2), 139 - 44
The relationship between HP1 and S2 bacteriophages of Haemophilus influenzae; Skowronek K et al.; Comparison of the nucleotide sequences of the left arms of two Haemophilus influenzae phages, S2 and HP1 is presented . They exhibit a characteristic mosaic pattern of homologous and non-homologous regions . The homology extends over the attP site and int, orf 5 to 9, rep and the 3' part of cI genes . Two major non-homologous regions were detected . One is found between the int and cI genes; the other spans the region of promoters and the cox gene . Variations in the region of the promotors which is involved in the choice between a lysogenic and a lytic pathway and some divergences in the cI coding sequences are probably responsible for the observed immunity differences between the two phages . Distinctions in the distribution of consensus sequences for an integration host factor (IHF) and integrase-binding sites and promoters are described . These data offer an explanation of the relationship between three types of S2/HP1 phages . It allows in turn a final settlement of the nomenclature variation in the literature . The results presented, which are similar to those obtained for other phage groups, suggest that the mosaic structure of phage genomes is a normal outcome of phage divergence.

Nucleic Acids Res, 1997 Oct 15, 25(20), 4147 - 52
The Neisseria gonorrhoeae S.NgoVIII restriction/modification system: a type IIs system homologous to the Haemophilus parahaemolyticus HphI restriction/modification system; Gunn JS et al.; Strains of Neisseria gonorrhoeae possess numerous restriction-modification (R-M) systems . One of these systems, which has been found in all strains tested, encodes the S . NgoVIII specificity (5'TCACC 3') R-M system . We cloned two adjacent methyltransferase genes (dcmH and damH), each encoding proteins whose actions protect DNA from digestion by R.HphI or R.Ngo BI (5'TCACC 3') . The damH gene product is a N 6-methyladenine methyltransferase that recognizes this sequence . We constructed a plasmid containing multiple copies of the S.NgoVIII sequence, grew it in the presence of damH and used the HPLC to demonstrate the presence of N 6-methyladenine in the DNA . A second plasmid, containing overlapping damH and Escherichia coli dam recognition sequences in combination with various restriction digests, was used to identify which adenine in the recognition sequence was modified by damH . The predicted dcmH gene product is homologous to 5-methylcytosine methyltransferases . The products of both the dcmH and damH genes, as well as an open reading frame downstream of the damH gene are highly similar to the Haemophilus parahaemolyticus hphIMC , hphIMA and hphIR gene products, encoding the Hph I Type IIs R-M system . The S.NgoVIII R-M genes are flanked by a 97 bp direct repeat that may be involved in the mobility of this R-M system.

Infect Immun, 1997 Oct, 65(10), 4250 - 7
Haemophilus somnus immunoglobulin binding proteins and surface fibrils; Corbeil LB et al.; The high-molecular-weight (HMW) immunoglobulin binding proteins (IgBPs) of Haemophilus somnus and a 76-kDa surface protein (p76) are found in serum-resistant virulent strains but not in several serum-sensitive strains from asymptomatic carriers . For the first time, p76 was shown to be an IgBP also . This was done by competitive inhibition studies with affinity-purified antidinitrophenol (anti-DNP) and DNP to ensure that binding was not antigen specific . The HMW IgBPs, but not the p76 IgBP, were partially purified from concentrated culture supernatant in detergent by fluid-phase liquid chromatography with a gel filtration column . Membrane extraction studies showed that p76 predominated in the Sarkosyl-soluble fraction of the bacterial cell pellet . Since integral outer membrane (OM) proteins are Sarkosyl insoluble, this is consistent with our previous finding that implicated p76 as a peripheral OM protein . The HMW IgBPs were found predominantly in the Sarkosyl-soluble fraction of the culture supernatant . This suggests that they were not integral membrane proteins and that their presence in the supernatant was not due to OM blebbing . We then showed that two IgBP-positive serum-resistant virulent strains have a surface fibrillar network but that two IgBP-negative serum-sensitive H . somnus strains from asymptomatic preputial carriers do not . Fibrils on the surfaces of IgBP+ strains bound gold-labelled bovine immunoglobulin G2 (IgG2) anti-DNP, indicating that these fibrils have IgG2 binding activity . Therefore, this study shows that H . somnus has two IgBPs, including a peripheral membrane protein and a fibrillar surface network.

Mol Microbiol, 1997 Aug, 25(4), 619 - 37
Comparison of archaeal and bacterial genomes: computer analysis of protein sequences predicts novel functions and suggests a chimeric origin for the archaea; Koonin EV et al.; Protein sequences encoded in three complete bacterial genomes, those of Haemophilus influenzae, Mycoplasma genitalium and Synechocystis sp., and the first available archaeal genome sequence, that of Methanococcus jannaschii, were analysed using the BLAST2 algorithm and methods for amino acid motif detection . Between 75% and 90% of the predicted proteins encoded in each of the bacterial genomes and 73% of the M . jannaschii proteins showed significant sequence similarity to proteins from other species . The fraction of bacterial and archaeal proteins containing regions conserved over long phylogenetic distances is nearly the same and close to 70% . Functions of 70-85% of the bacterial proteins and about 70% of the archaeal proteins were predicted with varying precision . This contrasts with the previous report that more than half of the archaeal proteins have no homologues and shows that, with more sensitive methods and detailed analysis of conserved motifs, archaeal genomes become as amenable to meaningful interpretation by computer as bacterial genomes . The analysis of conserved motifs resulted in the prediction of a number of previously undetected functions of bacterial and archaeal proteins and in the identification of novel protein families . In spite of the generally high conservation of protein sequences, orthologues of 25% or less of the M . jannaschii genes were detected in each individual completely sequenced genome, supporting the uniqueness of archaea as a distinct domain of life . About 53% of the M . jannaschii proteins belong to families of paralogues, a fraction similar to that in bacteria with larger genomes, such as Synechocystis sp . and Escherichia coli, but higher than that in H . influenzae, which has approximately the same number of genes as M . jannaschii . Certain groups of proteins, e.g . molecular chaperones and DNA repair enzymes, thought to be ubiquitous and represented in the minimal gene set derived by bacterial genome comparison, are missing in M . jannaschii, indicating massive non-orthologous displacement of genes responsible for essential functions . An unexpectedly large fraction of the M . jannaschii gene products, 44%, shows significantly higher similarity to bacterial than to eukaryotic proteins, compared with 13% that have eukaryotic proteins as their closest homologues (the rest of the proteins show approximately the same level of similarity to bacterial and eukaryotic homologues or have no homologues) . Proteins involved in translation, transcription, replication and protein secretion are most closely related to eukaryotic proteins, whereas metabolic enzymes, metabolite uptake systems, enzymes for cell wall biosynthesis and many uncharacterized proteins appear to be 'bacterial' . A similar prevalence of proteins of apparent bacterial origin was observed among the currently available sequences from the distantly related archaeal genus, Sulfolobus . It is likely that the evolution of archaea included at least one major merger between ancestral cells from the bacterial lineage and the lineage leading to the eukaryotic nucleocytoplasm.

J Periodontol, 1997 Sep, 68(9), 842 - 50
The effect of smoking on serum IgG2 reactive with Actinobacillus actinomycetemcomitans in early-onset periodontitis patients; Tangada SD et al.; High titers of serum IgG2 reactive with Actinobacillus actinomycetemcomitans are present in early-onset periodontitis (EOP) patients and it appears that anti-A . actinomycetemcomitans may be protective . Smoking is associated with increased periodontal disease severity in generalized early-onset periodontitis (G-EOP) patients, but is not associated with periodontal disease severity in patients with localized juvenile periodontitis (LJP) . Furthermore, smoking is associated with reduced serum IgG2 levels in black patients with G-EOP but not in those with LJP . Based on this selective effect of smoking, we hypothesized that smoking would be associated with a reduction of specific IgG2 reactive with A . actinomycetemcomitans in black G-EOP patients but not black LJP patients . In addition, we examined IgG2 responses to carbohydrate antigens from non-periodontal pathogens including Haemophilus influenzae b oligosaccharide antigen (Hib) and the Streptococcus pneumoniae antigen phosphocholine (PC) . Smoking status was assessed from serum cotinine levels, and IgG2 specific for A . actinomycetemcomitans, Hib, and PC was assessed by ELISA . Our study revealed that smoking was correlated with a dramatic reduction in serum IgG2 anti-A . actinomycetemcomitans in G-EOP smokers but not in LJP smokers . In contrast, anti-Hib IgG2 and anti-PC IgG2 were not affected in either G-EOP or LJP patients . In short, these results indicate that smoking is associated with a reduction in serum IgG2 anti-A . actinomycetemcomitans in black G-EOP subjects, but IgG2 reactive with other antigens may not be reduced in G-EOP smokers.

Clin Infect Dis, 1997 Sep, 25(3), 600 - 6
Antibody response to Haemophilus influenzae type b vaccine in relation to the number of CD4+ T lymphocytes in adults infected with human immunodeficiency virus; Kroon FP et al.; A prospective study of antibody production by adults infected with human immunodeficiency virus (HIV) after vaccination with tetanus toxoid-conjugated Haemophilus influenzae type b (Hib) vaccine was conducted . Concentrations of antibodies to the two immunogenic components of the vaccine (i.e., polyribosylribitolphosphate {PRP} and tetanus toxin) were determined . Individuals were divided into three groups according to the CD4+ T lymphocyte count: group 1, < or =100 x 10(6)/L; group 2, 101-300 x 10(6)/L; and group 3, >300 x 10(6)/L . After vaccination, concentrations of IgM and IgG antibodies to PRP were significantly lower in group 1 than in the other patient groups and controls . A CD4+ T lymphocyte count of <100 x 10(6)/L and an impaired proliferative response of lymphocytes to monoclonal antibody to CD3 were independently associated with a less than threefold increase in concentrations of IgG antibody to PRP . Analysis of IgG subclasses demonstrated that the production of IgG1 antibodies was predominantly affected . Postvaccination concentrations of antibody to tetanus toxin were significantly lower in group 1 than in group 3 and controls . Both prevaccination and postvaccination concentrations of antibody to tetanus toxin were not correlated with the magnitude of the response of antibody to PRP . We conclude that HIV-infected individuals with CD4+ T Iymphocyte counts of <100 x 10(6)/L demonstrate an impaired antibody response after vaccination with conjugated Hib vaccine.

J Virol, 1997 Oct, 71(10), 7579 - 85
Cleavage of influenza A virus H1 hemagglutinin by swine respiratory bacterial proteases; Callan RJ et al.; Cleavage of influenza A virus hemagglutinin (HA) is required for expression of fusion activity and virus entry into cells . Extracellular proteases are responsible for the proteolytic cleavage activation of avirulent avian and mammalian influenza viruses and contribute to pathogenicity and tissue tropism . The relative contributions of host and microbial proteases to cleavage activation in natural infection remain to be established . We examined 23 respiratory bacterial pathogens and 150 aerobic bacterial isolates cultured from the nasal cavities of pigs for proteolytic activity . No evidence of secreted proteases was found for the bacterial pathogens, including Haemophilus parasuis, Pasteurella multocida, Actinobacillus pleuropneumoniae, Bordetella bronchiseptica, and Streptococcus suis . Proteolytic bacteria were isolated from 7 of 11 swine nasal samples and included Staphylococcus chromogenes, Staphylococcus hyicus, Aeromonas caviae, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Enterococcus sp . Only P . aeruginosa secreted a protease, elastase, that cleaved influenza virus HA . However, compared to trypsin, the site of cleavage by elastase was shifted one amino acid in the carboxy-terminal direction and resulted in inactivation of the virus . Under the conditions of this study, we identified several bacterial isolates from the respiratory tracts of pigs that secrete proteases in vitro . However, none of these proteolytic isolates demonstrated direct cleavage activation of influenza virus HA.

FEMS Microbiol Lett, 1997 Sep 15, 154(2), 311 - 6
Importance of the galE gene on the virulence of Pasteurella multocida; Fernandez de Henestrosa AR et al.; The galE gene of Pasteurella multocida has been isolated by complementing galE-defective mutants of Salmonella typhimurium with a plasmid library of this organism . The complete nucleotide sequence of the P . multocida galE gene consists of 1017 nucleotides, encoding a predicted polypeptide of 339 amino acids . The deduced amino acid sequence displayed the highest identity (85%) to the GalE protein of Haemophilus influenzae . However, the gene organization surrounding the galE locus was different from that of H . influenzae . A galE-defective mutant of P . multocida was obtained by replacement of the active galE gene by a copy inactivated in vitro . The resulting galE mutant was highly attenuated as seen in a biological test carried out in a mouse model.

FEMS Microbiol Lett, 1997 Sep 15, 154(2), 303 - 10
Identification of the Treponema pallidum subsp . pallidum glycerophosphodiester phosphodiesterase homologue; Stebeck CE et al.; To identify potential opsonic targets of Treponema pallidum subsp . pallidum, a treponemal genomic expression library was constructed and differentially screened with opsonic and non-opsonic T . pallidum antisera . This method identified an immunoreactive clone containing an open reading frame encoding a 356 residue protein . Nucleotide sequence analysis demonstrated the translated protein to be a homologue of glycerophosphodiester phosphodiesterase, a glycerol metabolizing enzyme previously identified in Haemophilus influenzae, Escherichia coli, Bacillus subtilis and Borrelia hermsii . Sequence alignment analyses revealed the T . pallidum and H . influenzae enzymes share a high degree of amino acid sequence similarity (72%), suggesting that in T . pallidum this molecule may be surface exposed and involved in IgD binding as is the case with its counterpart in H . influenzae.

FEMS Microbiol Lett, 1997 Sep 15, 154(2), 251 - 8
Transformation of a virulence associated gene of Haemophilus somnus into a strain lacking the gene; Sanders JD et al.; The role of a 76 kDa surface antigen (p76) of Haemophilus somnus in virulence was investigated . The p76 gene from a virulent isolate of H . somnus (strain 2336) was introduced into an asymptomatic carrier strain (129Pt) lacking this gene . This was accomplished by the development of a system for genetic exchange in H . somnus . The cloned p76 gene was inserted into the broad host range vector pLS88, electroporated into H . influenzae for modification and then into the H . somnus strain 129Pt . The recombinant plasmid was characterized from selected transformants and expression of the p76 protein was demonstrated by Western immunoblotting . However, transformants were not serum resistant and surface exposure of the recombinant protein could not be detected, suggesting that additional genetic elements might be required for export.

J Pediatr Ophthalmol Strabismus, 1997 Sep-Oct, 34(5), 293 - 6
Periorbital and orbital cellulitis in the Haemophilus influenzae vaccine era; Barone SR et al.; BACKGROUND: Prior to development of the Haemophilus-influenzae vaccine, a significant number of periorbital and orbital infections were associated with H influenzae type b bacteremia . The incidence of invasive H influenzae type b disease has decreased dramatically since the introduction of these vaccines . The effect of the H influenzae type b vaccine on the microbiology of periorbital and orbital infections has not been examined . METHODS: Charts of 134 hospitalized patients with the discharge diagnosis of periorbital or orbital cellulitis from 1985 to 1995 were reviewed . RESULTS: In contrast with studies performed in the pre-vaccine era, H influenzae type b was not a significant pathogen . The pathogen was isolated from only 2 of 133 blood cultures performed, and was not isolated from any of the 101 patients discharged after July 1987 . Sinusitis, however, remains an important cause of these eye infections . Of patients who underwent radiographic evaluation, sinusitis was present in 96% of children with orbital cellulitis and 81% of patients with periorbital cellulitis . CONCLUSIONS: H influenzae type b is no longer a significant pathogen in periorbital or orbital cellulitis and management should be modified accordingly.

Behring Inst Mitt, 1997 Feb, (98), 191 - 6
Bacterial ghosts as multifunctional vaccine particles; Szostak MP et al.; Expression of cloned PhiX174 gene E in Gram-negative bacteria results in lysis of the bacteria by formation of an E-specific transmembrane tunnel structure built through the cell envelope complex . Bacterial ghosts have been produced from a variety of bacteria including Escherichia coli . Salmonella typhimurium, Salmonella enteritidis, Vibrio cholerae, Klebsiella pneumoniae, Actinobacillus pleuropneumoniae, Haemophilus influenzae, Pasteurella haemolytica, Pasteurella multocida, and Helicobacter pylori . Such ghosts are used as non-living candidate vaccines and represent an alternative to heat or chemically inactivated bacteria . In recombinant ghosts, foreign proteins can be inserted into the inner membrane prior to E-mediated lysis via specific N-, or C-, or N- and C-terminal anchor sequences . The export of proteins into the periplasmic space or the expression of recombinant S-layer proteins vastly extents the capacity of ghosts or recombinant ghosts as carriers of foreign epitopes or proteins . Oral, aerogenic or parenteral applications of (recombinant) ghosts in experimental animals induced specific humoral and cellular immune responses against bacterial and target components including protective mucosal immunity . The most relevant advantage of ghosts and recombinant bacterial ghosts as immunogens is that no inactivation procedures that denature relevant immunogenic determinants are employed in the production of ghosts used as vaccines or as carriers of relevant antigens . The inserted target antigens into the inner membrane or into S-layer proteins are not limited in size.

Antimicrob Agents Chemother, 1997 Sep, 41(9), 1991 - 5
Pharmacokinetics of florfenicol in cerebrospinal fluid and plasma of calves; de Craene BA et al.; Florfenicol, a fluorinated analog of thiamphenicol, is of great value in veterinary infectious diseases that formerly responded favorably to chloramphenicol . In view of the treatment of meningitis in calves, we studied its pharmacokinetics in the cerebrospinal fluid (CSF) and plasma of six animals . To this end, a new high-performance liquid chromatography method was developed which, unlike previous ones, uses solid-phase instead of double-phase extraction to isolate the drug . After a single intravenous dose of 20 mg/kg of body weight, a maximum concentration in CSF of 4.67 +/- 1.51 microg/ml (n = 6) was reached, with a mean residence time of 8.7 h . The decline of florfenicol in both CSF and plasma fitted a biexponential model with elimination half-lives of 13.4 and 3.2 h, respectively . Florfenicol penetrated well into CSF, as evidenced from an availability of 46% +/- 3% relative to plasma . The levels remained above the MIC for Haemophilus somnus over a 20-h period . Our results provide evidence indicating the effectiveness of florfenicol in the treatment of bacterial meningitis of calves.

Antimicrob Agents Chemother, 1997 Sep, 41(9), 1979 - 84
Amoxicillin treatment of experimental acute otitis media caused by Haemophilus influenzae with non-beta-lactamase-mediated resistance to beta-lactams: aspects of virulence and treatment; Melhus A et al.; Through alterations primarily in the penicillin-binding proteins, a non-beta-lactamase-mediated resistance to beta-lactams has evolved in Haemophilus influenzae . The virulence of these chromosomally changed strains has been questioned . To ascertain whether these alterations involve a reduction in virulence of H . influenzae and whether they could be advantageous for the bacterium during amoxicillin treatment of acute otitis media, a total of 70 Sprague-Dawley rats were challenged with a susceptible recipient strain or a genetically similar resistant transformant strain . Antibiotic therapy was started on day 3 after inoculation, and the animals were monitored by daily otomicroscopy and analysis of bacterial samples from middle ear effusions obtained on day 8, the last day of observation . The animals were also sacrificed on days 4 and 8 and after 2 months for morphological examination . Compared with the susceptible recipient strain, recovery from infections caused by the resistant transformant strain was delayed, and the late structural changes were more severe in the animals challenged with the latter strain . The results of the study indicate that chromosomal alterations mediating a relatively low level of resistance to beta-lactams may be advantageous for H . influenzae during antibiotic treatment of a local infection in the rat, and the alterations may occur without any significant loss of virulence.

Antimicrob Agents Chemother, 1997 Sep, 41(9), 1965 - 72
A multicenter, randomized study comparing the efficacy and safety of intravenous and/or oral levofloxacin versus ceftriaxone and/or cefuroxime axetil in treatment of adults with community-acquired pneumonia; File TM Jr et al.; Five hundred ninety patients were enrolled in a prospective, multicenter, randomized trial comparing the efficacy and safety of 7 to 14 days of levofloxacin treatment with that of ceftriaxone and/or cefuroxime axetil in the management of community-acquired pneumonia in adults . Patients received either intravenous and/or oral levofloxacin (500 mg once daily) or the comparative agents, parenteral ceftriaxone (1 to 2 g once to twice daily) and/or oral cefuroxime axetil (500 mg twice daily) . Erythromycin or doxycycline could be added to the comparator arm at the investigator's discretion . The decision to use an intravenous or oral antimicrobial agent for initial therapy was made by the investigator . Clinical and microbiological evaluations were completed at the baseline, during treatment, 5 to 7 days posttherapy, and 3 to 4 weeks posttherapy . Four hundred fifty-six patients (226 given levofloxacin and 230 administered ceftriaxone and/or cefuroxime axetil) were evaluable for clinical efficacy . Streptococcus pneumoniae and Haemophilus influenzae were isolated in 15 and 12%, respectively, of clinically evaluable patients . One hundred fifty atypical pathogens were identified: 101 were Chlamydia pneumoniae, 41 were Mycoplasma pneumoniae, and 8 were Legionella pneumophila . Clinical success at 5 to 7 days posttherapy was superior for the levofloxacin group (96%) compared with the ceftriaxone and/or cefuroxime axetil group (90%) (95% confidence interval {CI} of -10.7 to -1.3) . Among patients with typical respiratory pathogens who were evaluable for microbiological efficacy, the overall bacteriologic eradication rates were superior for levofloxacin (98%) compared with the ceftriaxone and/or cefuroxime axetil group (85%) (95% CI of -21.6 to -4.8) . Levofloxacin eradicated 100% of the most frequently reported respiratory pathogens (i.e., H . influenzae and S . pneumoniae) and provided a >98% clinical success rate in patients with atypical pathogens . Both levofloxacin and ceftriaxone-cefuroxime axetil eradicated 100% of the S . pneumoniae cells detected in blood culture . Drug-related adverse events were reported in 5.8% of patients receiving levofloxacin and in 8.5% of patients administered ceftriaxone and/or cefuroxime axetil . Gastrointestinal and central and peripheral nervous system adverse events were the most common events reported in each treatment group . In conclusion, these results demonstrate that treatment with levofloxacin is superior to ceftriaxone and/or cefuroxime axetil therapy in the management of community-acquired pneumonia in adults.

JAMA, 1997 Sep 17, 278(11), 925 - 31
Dexamethasone as adjunctive therapy in bacterial meningitis . A meta-analysis of randomized clinical trials since 1988; McIntyre PB et al.; OBJECTIVE: To evaluate the effectiveness of dexamethasone in bacterial meningitis in the subcategories of causative organism and timing and nature of antibiotic therapy . DATA SOURCES: MEDLINE, HEALTHLINE, and AIDSLINE were searched with the Medical Subject Headings "dexamethasone" and "meningitis" in any language . Bibliographies, conference abstracts, and the authors of identified studies were consulted . STUDY SELECTION: Randomized, concurrently controlled trials of dexamethasone therapy in childhood bacterial meningitis published from 1988 to November 1996 were selected . Of 16 studies identified, 5 were not eligible . DATA EXTRACTION: Data were extracted by means of standard outcomes in a protocol sent to all principal authors . DATA SYNTHESIS: Random-effects meta-analysis models were used to obtain summary estimates . As the incidence of severe hearing loss differed significantly by organism among control subjects, organism-specific estimates were used . In Haemophilus influenzae type b meningitis, dexamethasone reduced severe hearing loss overall (combined odds ratio {OR}, 0.31; 95% confidence interval {CI}, 0.14-0.69) . Similar ORs were obtained after studies were stratified by the timing of administration of dexamethasone (before or with antibiotics vs later) or by type of antibiotic (cefuroxime vs other) . In pneumococcal meningitis, only studies in which dexamethasone was given early suggested protection, which was significant for severe hearing loss (combined OR, 0.09; 95% CI, 0.0-0.71) and approached significance for any neurological or hearing deficit (combined OR, 0.23; 95% CI, 0.04-1.05) . For all organisms combined, the pooled OR suggested protection against neurological deficits other than hearing loss but was not significant (OR, 0.59; 95% CI, 0.34-1.02) . Outcomes were similar in studies that used 2 vs more than 2 days of dexamethasone therapy . Adverse effects were not significantly increased with dexamethasone except for secondary fever . The incidence of gastrointestinal tract bleeding increased with longer duration of dexamethasone treatment (0.5% in controls, 0.8% with 2 days of treatment, 3.0% with 4 days of treatment) . CONCLUSIONS: The available evidence on adjunctive dexamethasone therapy confirms benefit for H influenzae type b meningitis and, if commenced with or before parenteral antibiotics, suggests benefit for pneumococcal meningitis in childhood . Limiting dexamethasone therapy to 2 days may be optimal.

J Antimicrob Chemother, 1997 Aug, 40(2), 179 - 87
Susceptibility testing of macrolide and lincosamide antibiotics according to DIN guidelines . Deutsches Institut für Normung; Schmalreck AF et al.; The in-vitro activity of azithromycin, clarithromycin, erythromycin, josamycin, midekamycin, roxithromycin and clindamycin against 674 Gram-negative and Gram-positive clinical isolates, including methicillin-resistant Staphylococcus aureus, was determined by agar dilution, microdilution and agar diffusion with Mueller-Hinton medium according to the Deutsches Institut fur Normung (DIN) 58940 guidelines . The results obtained by regression analysis and the error-rate-bounded method of Metzler-DeHaan indicate that common interpretative criteria (breakpoints) for test discs may be assigned to susceptible/resistant Gram-positive strains for all antibiotics tested . The following tentative DIN values are suggested for 15 microg macrolide discs: for susceptible Gram-positive and Gram-negative strains, an inhibition zone diameter (IZD) of > or = 26 mm at a corresponding MIC of < or = 2 mg/L; for resistant Gram-positive strains, an IZD of < or = 21 mm; for resistant Gram-negative strains, an IZD of < or = 19 mm at a corresponding MIC of > or = 8 mg/L . For Haemophilus influenzae only, breakpoints for azithromycin are suggested with IZDs of > or = 21 mm for susceptible and < or = 18 mm for resistant.

Electrophoresis, 1997 Aug, 18(8), 1472 - 82
Development of a Haemophilus two-dimensional protein database; Cash P et al.; Members of the Haemophilus genus are responsible for various human infections including respiratory infections and meningitis . The complete nucleotide sequence of the Rd strain of Haemophilus influenzae has been reported and represents a valuable resource to investigate gene expression within this bacterial group . We described previously the application of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) to characterise the proteins of Haemophilus influenzae (Cash et al., Electrophoresis 1995, 16, 135-148) . We have extended these data with comparative studies of the proteins from other members of the Haemophilus genus (specifically H . parainfluenzae, H . haemolyticus and H . parahaemolyticus) to identify homologous proteins and, by extension, the genes encoding them, among these bacteria . The proteins extracted from each of these bacterial isolates were compared by coelectrophoresis to the 2-D protein profile of the reference nontypable strain of H . influenzae (HI-64443) used as the basis for the 2-D protein database . A composite reference 2-D protein profile of HI-64443 was derived from three independent analyses of the soluble bacterial proteins . Between 21% and 37% of the HI-64443 proteins from the reference 2-D protein profile comigrated with proteins in the other isolates from the Haemophilus genus . This compared with 62% and 64% comigration when HI-64443 was compared with the Eagan and Rd strains of H . influenzae, respectively . The 2-D protein profile of the Rd strain of H . influenzae was compared to that of HI-64443 by coelectrophoresis; 64% of the proteins detected for the Rd strain comigrated with proteins found for HI-64443 when analysed in parallel . The capacity of 2-D PAGE to investigate global interactions of gene expression was applied to the analysis of superoxide dismutase (SOD) expression in H . influenzae strain Eagan . A "knock-out" mutant in the sodA gene which encodes {Mn}-SOD was characterised with respect to protein synthesis compared to the parental isolate . From these analyses, the primary product of sodA was provisionally identified as a protein with a molecular mass of 25500 Da and an estimated pI of 6.55 . Quantitative changes in the expression of two other proteins in the SOD mutant were detected by comparison with the parental isolate . These data are discussed in relation to the development of a 2-D protein database for H . influenzae and related bacteria to investigate genome homologies and gene expression.

Electrophoresis, 1997 Aug, 18(8), 1410 - 7
Evaluation of algorithms used for cross-species proteome characterisation; Cordwell SJ et al.; The ability to effectively search databases for the identification of protein spots from two-dimensional electrophoresis gels has become an essential step in the study of microbial proteomes . A variety of analytical techniques are currently being employed during protein characterisation . A number of algorithms used to search databases, accessible via the World Wide Web, depend upon information concerning N- and C-terminal microsequence, amino acid composition, and peptide-mass fingerprinting . The effectiveness of nine such algorithms, as well as COMBINED (software developed in this laboratory for identifying proteins across species boundaries) was examined . Fifty-four ribosomal proteins from the Mycoplasma genitalium genome, and 72 amino acyl tRNA synthetases from the Haemophilus influenzae, M . genitalium and Methanococcus jannaschii genomes were chosen for study . These proteins were selected because they represent a wide range of sequence identities across species boundaries (22.7-100% identity), as detected by standard sequence alignment tools . Such sequence variation allowed for a statistical comparison of algorithm success measured against published sequence identity . The ability of analytical techniques used in protein characterisation and associated database query programs to detect identity at the functional group level was examined for proteins with low levels of homology at the gene/protein sequence level . The significance of these theoretical data manipulations provided the means to predict the utility of data acquired experimentally for non-sequence-dependent software in proteome analysis . The data obtained also predicted that 'sequence tagging' of peptide fingerprints would need to be accompanied by at least 11-20 residues of amino acid sequence for it to be widely used for protein characterisation across species boundaries.

Electrophoresis, 1997 Aug, 18(8), 1314 - 34
Identifying the major proteome components of Haemophilus influenzae type-strain NCTC 8143; Link AJ et al.; With the completion of the Haemophilus influenzae Rd genomic sequence, we know the identity of most of the theoretical proteins in the proteome of this bacterium . However, the most abundant components of the actual proteome are unknown . Using mass spectrometry and two-dimensional gel electrophoresis (2-DE), we sequenced and analyzed the most abundant proteins observed in the ATCC reference strain of H . influenzae, NCTC 8143 (303 of approximately 400 Coomassie-stained 2-DE spots) . To automate the identification of 2-DE spots, we coupled a liquid autosampler to a microcolumn liquid chromatography electrospray ionization tandem mass spectrometer capable of identifying 22 spots per day . From the 303 sequenced spots, we identified 263 unique proteins . Most of the abundant proteins lie in an isoelectric point range of pH 4-7 and a molecular mass range of 10-100 kDa . Of the observed proteins, the most abundant is the outer membrane protein P2 . Based on variety and abundance, proteins involved in energy metabolism and macromolecular synthesis are the dominant classes of proteins . Unexpectedly, tryptophanase was identified as a highly abundant protein in the strain NCTC 8143 whose sequence is not present in the genome of the Rd strain . By searching the tandem mass spectra against the translated genomic sequence, we identified several proteins which were not annotated in the genomic sequence . Surprisingly, 22% of the identified 2-DE spots represent isoforms in which gene products with the same primary sequence have different observed pI and M(r), indicating that these proteins are post-translationally processed . Although most proteins' predicted and observed isoelectric points and molecular masses show reasonable concordance, the observed values for several proteins deviate significantly from the predicted values . These anomalies may represent either highly processed proteins or misinterpretations of the genomic sequence . Using the technology developed in this project, the protein expression of other strains of H . influenzae grown under different environmental conditions can be compared to identify differences in their proteomes.

Arch Ophthalmol, 1997 Sep, 115(9), 1124 - 8
Outpatient treatment of bleb infection; Chen PP et al.; OBJECTIVE: To determine the outcome of outpatient treatment of bleb infection after trabeculectomy . DESIGN: Retrospective review . SETTING: University referral center . PATIENTS: Twelve eyes with bleb infection and without clinical vitreous involvement of 11 patients treated as out-patients from January 1994 to May 1996 . INTERVENTIONS: After culture of the bleb surface, treatment consisted of intensive topical antibiotics alone (9 eyes), combined with subconjunctival antibiotic injection (1 eye), oral antibiotics (1 eye), or both (1 eye) . Five eyes received topical corticosteroids . MAIN OUTCOME MEASURES: Visual acuity, intraocular pressure, and bleb characteristics . RESULTS: All eyes responded to treatment within 24 to 48 hours, and recovered visual acuity to within 1 line of the preinfection level . Intraocular pressure continued to be controlled after resolution of infection in 11 eyes (92%); 1 bleb failed and medical therapy was resumed . Bleb leaks were noted during treatment of infection in 11 eyes, and 5 eyes (42%) underwent bleb revision . Organisms cultured from the bleb surface included Staphylococcus aureus (8 eyes), Haemophilus influenzae (2 eyes), and Streptococcus (1 eye) . Outpatient treatment costs were estimated to be 78% less than inpatient treatment costs, per 24 hours . CONCLUSION: Outpatient treatment of bleb infection was efficacious in this group of patients.

Lijec Vjesn, 1997 Feb, 119(2), 72 - 6
{New vaccines for wide usage}; Mravunac B et al.; In the introduction achievements of obligatory applied vaccines are described . Data on new vaccines for wide application are presented: acellular pertussis vaccine, Haemophilus influenzae b vaccine, hepatitis B vaccine and varicella (zoster) vaccine . For each vaccine data on immunity, protection and side effects are presented . Indications (epidemiological, illness severity) justifying vaccination as a method of protection from infection with a distinctive causative agent are presented . Antigen structure is given for each vaccine . Finally the form of application and age of primovaccination and revaccination are given . The conclusion is that these vaccines give high immunity and protection like those already in wide (obligatory) usage, and have less side effects.

Ann Pharmacother, 1997 Sep, 31(9), 1022 - 33
Ceftibuten: a new expanded-spectrum oral cephalosporin; Guay DR; OBJECTIVE: To review the antimicrobial activity, pharmacokinetics, clinical efficacy, and tolerability of ceftibuten, a new expanded-spectrum oral cephalosporin . DATA SOURCES: Literature was identified by a MEDLINE search (January 1983-June 1996) of the medical literature, review of English-language literature and bibliographies of these articles, and data on file . STUDY SELECTION: Clinical efficacy data were selected from all published and unpublished trials and abstracts that mentioned ceftibuten . Additional information concerning in vitro susceptibility, safety, chemistry, and pharmacokinetic profile of ceftibuten also was reviewed . DATA SYNTHESIS: Ceftibuten, an oral expanded-spectrum cephalosporin, has a broad spectrum of activity against many gram-negative and selected gram-positive organisms, including Streptococcus pneumoniae, Streptococcus pyogenes, Moraxella catarrhalis, and Haemophilus influenzae . Ceftibuten is stable to hydrolysis by many common beta-lactamases . Ceftibuten is rapidly and almost completely absorbed from the gastrointestinal tract and is primarily eliminated renally as unchanged drug . The elimination half-life of ceftibuten is slightly longer than 2 hours . Efficacy has been demonstrated in a number of clinical trials in adults and children with upper and lower respiratory tract infections (e.g., acute otitis media, pharyngitis, sinusitis, bronchitis) and urinary tract infections . The adverse effect profile is equal to that of comparator agents . CONCLUSIONS: Ceftibuten is an alternative to other antimicrobial agents with convenient once-daily dosing in the treatment of upper and lower respiratory tract infections . Similar to other oral expanded-spectrum cephalosporins, ceftibuten has antimicrobial activity against common pathogens of the respiratory tract and is stable in the presence of many beta-lactamases . The clinical choice of an oral expanded-spectrum cephalosporin will be based on patient acceptance, frequency of administration, and cost.

Rev Pneumol Clin, 1997, 53(3), 138 - 43
{Bacterial infectious agents implicated in lower respiratory tract infections in general practice}; Vernejoux JM et al.; The consensus of the French Society of Infectious Diseases established in 1991 states that Streptococcus pneumoniae and Haemophilus influenzae are the main causal agents of community-acquired lower airway infections and that antibiotics constitute the "prudent" solution in case of acute bronchitis which persists more than one week or in case of pneumonia in "fragile" at-risk adults . The efficacy of these "probabilistic" recommendations depends on the epidemiology of the infectious agents . The objective of this study was to identify the causal germs in lower airway infections and determine their sensitivity to the antibiotics recommended in the consensus statement . The study was conducted from December to March, in 1992 and 1993 . Expectoration samples were obtained from 111 cases including 29 patients with chronic bronchitis . Seventy different strains were isolated including 24 strains of H . influenzae (3 betalactamase producers), 15 strains of S . pneumoniae (1 with reduced sensitivity to peni G: MIC = 1 mu/ml), 9 strains of S . aureus (2 methicillin resistant), and 8 strains of Branhamella catarrhalis (6 betalactamase producers) . The number of positive serologies was very low: 5 Chlamydiae pneumoniae, 2 Chlamydiae trachomatis and 1 Mycoplasma pneumoniae . In conclusion H . influenzae is the most frequent germ; S . pneumoniae infections with reduced peni-G sensitivity and atypical germs are uncommon . The consensus recommendations appear to be adapted to the bacterial flore causing community-acquired lower airway infection in healthy and at-risk subjects.

Diagn Microbiol Infect Dis, 1997 Jul, 28(3), 105 - 12
Evaluation of Haemophilus influenzae isolates with elevated MICs to amoxicillin/clavulanic acid; Jacobs MR et al.; A 1994 to 1995 national Haemophilus influenzae surveillance study of 1910 strains showed that 13 strains (0.7%) were resistant to amoxicillin/clavulanic acid (MIC, > or = 8/4 micrograms/ml) . These and other selected strains were investigated further in this study . Susceptibility of the surveillance study strains was determined with the commercial microdilution trays used in the original study and in triplicate with reference broth microdilution trays prepared by the investigators, as well as by Etest and disk diffusion . Amoxicillin/clavulanic acid resistance was confirmed for only one of the surveillance study strains . This strain produced double zones of growth with Etest and disk-diffusion methods, with the double zone containing spheroplasts . When the amoxicillin/clavulanic acid MICs of all beta-lactamase positive strains were compared, MIC results obtained with surveillance study trays and the Etest were one to two dilutions higher than MICs obtained with reference trays . The distribution and modal amoxicillin/clavulanic acid MICs of beta-lactamase-positive and -negative strains was essentially the same for a comparison group of strains using reference trays, in contrast to a fourfold higher modal MIC for beta-lactamase-positive strains using surveillance study reagents and strains . These differences in MICs could be attributed to variations in inoculum, the presence of spheroplasts, and/or a difference in potency of amoxicillin and/or clavulanic acid in the tray and Etest reagents used . Methods for assessing the adequacy of the clavulanic acid content are not adequate, amoxicillin control values and a beta-lactamase-positive H . influenzae control strain are required.

J Immunol Methods, 1997 Jul 14, 205(2), 191 - 200
Detection of meningitis antigens in buffer and body fluids by ultrasound-enhanced particle agglutination; Jenkins P et al.; The standard test card agglutination of antibody-coated latex by Neisseria meningitidis . Streptococcus group B, Haemophilus influenzae type b and Streptococcus pneumoniae antigens has been compared with a technique involving local concentration of the coated latex in an ultrasonic standing wave . The detection of positive control antigen was enhanced, compared with the test-card procedure, over a 16 to 64 fold range on exposure to ultrasound . Sample filtration eliminated non-specific agglutination on ultrasonic exposure of latex in control serum, urine or concentrated urine . Tests of meningitis patient body fluids showed increased detection of antigen with ultrasound for CSF (11/14 > 7/14) serum (8/13 > 3/13) and concentrated urine (8/17 > 2/17) compared to test card assays . The ultrasound detection of antigen in serum or concentrated urine was comparable to that achieved with CSF on test cards . Serum dilution experiments showed that ultrasound could detect antigen in serum over a 1000 fold concentration range.

J Dent Res, 1997 Sep, 76(9), 1538 - 47
Extracellular 37-kDa antigenic protein from Actinobacillus actinomycetemcomitans induces TNF-alpha, IL-1 beta, and IL-6 in murine macrophages; Tani Y et al.; The extracellular antigens of Actinobacillus actinomycetemcomitans Y4 (serotype b) contain a 37-kDa protein which is a major target for IgGs from patients suffering from severe alveolar bone loss . Since the 37-kDa protein has not been studied sufficiently, our investigation focused on its characteristics, e.g., its localization, specificity, and whether it directly stimulates macrophages to produce cytokines . The 37-kDa protein was purified from the culture supernatant of the Y4 strain by means of chromatofocusing and gel filtration . The 37-kDa protein is a unique glycoprotein which forms immune complexes with monoclonal antibodies against rhamnose-fucose polysaccharide . Patients with A . actinomycetemcomitans-associated periodontitis had higher antibody titers to the purified 37-kDa protein than healthy subjects (p < 0.001) . Anti-37-kDa protein antibodies recognized a 37-kDa band in the cytosolic, ribosomal, and total membrane fractions from Y4 cells . Extracellular substances from other strains of A . actinomycetemcomitans (serotypes a and c) also reacted in the Western blots, but Haemophilus spp . or several periodontopathic bacteria did not . These results suggested that the 37-kDa protein is a cytosolic protein that is passed through the cell membrane, and its protein portion is specific for A . actinomycetemcomitans but common to serotypes . This protein induced Il-1 beta, Il-6, and TNF-alpha release from murine macrophages . The Il-6-inducing activity of the 37-kDa protein was higher than that of LPS . These findings suggested that the 37-kDa protein which is released from live cells plays a role in A . actinomycetemcomitans-associated periodontitis, as antigen inducing the release of inflammatory cytokines which are associated with alveolar bone loss.

Proc Natl Acad Sci U S A, 1997 Sep 16, 94(19), 10313 - 8
The identification of CD4+ T cell epitopes with dedicated synthetic peptide libraries; Hiemstra HS et al.; For a large number of T cell-mediated immunopathologies, the disease-related antigens are not yet identified . Identification of T cell epitopes is of crucial importance for the development of immune-intervention strategies . We show that CD4+ T cell epitopes can be defined by using a new system for synthesis and screening of synthetic peptide libraries . These libraries are designed to bind to the HLA class II restriction molecule of the CD4+ T cell clone of interest . The screening is based on three selection rounds using partial release of 14-mer peptides from synthesis beads and subsequent sequencing of the remaining peptide attached to the bead . With this approach, two peptides were identified that stimulate the beta cell-reactive CD4+ T cell clone 1c10, which was isolated from a newly diagnosed insulin-dependent diabetes mellitus patient . After performing amino acid-substitution studies and protein database searches, a Haemophilus influenzae TonB-derived peptide was identified that stimulates clone 1c10 . The relevance of this finding for the pathogenesis of insulin-dependent diabetes mellitus is currently under investigation . We conclude that this system is capable of determining epitopes for (autoreactive) CD4+ T cell clones with previously unknown peptide specificity . This offers the possibility to define (auto)antigens by searching protein databases and/or to induce tolerance by using the peptide sequences identified . In addition the peptides might be used as leads to develop T cell receptor antagonists or anergy-inducing compounds.

Int J STD AIDS, 1997 Sep, 8(9), 585 - 8
Immunohistochemical investigations of genital ulcers caused by Haemophilus ducreyi; Abeck D et al.; To gain information on the specific composition of the inflammatory infiltrate of genital ulcers caused by Haemophilus ducreyi, biopsies of 6 genital ulcers which were diagnosed as chancroid on clinical and microbiological grounds were subjected to immunohistochemical investigations after conventional haematoxylineosin staining . A variety of antibodies reactive against B- and T-cells, plasma cells and granulocytes were used with each tissue sections . The lymphocytic infiltrate of chancroid ulcers consisted of both B- and T-lymphocytes and showed a cluster-like formation . B-lymphocytes were preferentially localized perivascularly in the middle layer, T-lymphocytes mainly in the deep layer of the inflamed oedematous tissue . Results stress the importance of both B- and T-cell mediated immune responses in Haemophilus ducreyi infectionPIP: Chancroid, the most prevalent form of genital ulcer disease in developing countries, increases the risk of HIV transmission . Use of monoclonal antibodies against leukocyte differentiation antigens enabled analysis of the composition of the inflammatory infiltrate of genital ulcers . In this study, biopsies of six genital ulcers caused by Haemophilus ducreyi were examined immunohistochemically . In each case, staining revealed a superficial necrotic layer of polymorphonuclear leukocytes with fibrin and erythrocytes at the base of the ulcer, a middle layer of the edematous inflammatory dermis with prominent blood vessels and vascular thrombi, and a deep layer of an inflammatory infiltrate of plasma cells and lymphocytes . The lymphocytic infiltrate of chancroid ulcers consisted of both B- and T-lymphocytes and showed a cluster-like formation . B-lymphocytes were preferentially localized perivascularly in the middle layer, while T-lymphocytes tended to be located in the deep layer of the inflamed edematous tissue . These findings provide further evidence of the importance of the involvement of T-cells in the local immune clearance of H . ducreyi . Future studies should investigate lymphocyte secretions detected in genital ulcers caused by H . ducreyi to gain more information on the role of the cellular immune mechanisms in the disease .

J Med Microbiol, 1997 Sep, 46(9), 763 - 72
Source of variation detected in ribotyping patterns of Haemophilus influenzae: comparison of traditional ribotyping, PCR-ribotyping and rDNA restriction analysis; Jordens JZ et al.; The pattern of EcoRI restriction fragments of chromosomal DNA that hybridize with a probe for genes encoding 16S and 23S rRNA is highly discriminatory for non-capsulate Haemophilus influenzae (NCHI) . The source of variation detected by these probe-based ribotyping patterns was investigated by restriction analysis of rRNA operon (rrn) amplification products from nine representative strains . Digestion of rrn amplification products with EcoRI indicated one conserved EcoRI site within 16S rDNA and no EcoRI sites within the 16S-23S intergenic spacer region of the nine strains, and an EcoRI site at the 5' end of 23S rDNA from seven of the nine strains . Comparison of the EcoRI ribotyping patterns obtained with separate probes for 16S and 23S rDNA showed more variation with the 23S probe indicating variation in EcoRI sites downstream from the operon . Restriction analyses of 16S and 23S rDNA amplification products with AluI, HhaI, HaeIII and TaqI divided the nine 'traditional' ribotypes into a maximum of three and eight groups, respectively . Similar analyses of the 16S-23S intergenic regions (PCR-ribotyping) failed to distinguish any of the nine representative strains . Therefore, there is probably insufficient variation within the operon for it to form a good target for PCR-based typing methods . In contrast, 'traditional' ribotyping with cDNA from 16S plus 23S rRNA detects restriction site differences in the sequences flanking the operon, which show considerably more variation between strains . 'Traditional' ribotyping should therefore remain the standard for characterising NCHI in epidemiological investigations.

Ann Allergy Asthma Immunol, 1997 Aug, 79(2), 145 - 50
Specific antibody responses to diphtheria/tetanus revaccination in children evaluated for immunodeficiency; McCusker C et al.; BACKGROUND: Assaying specific antibody levels against well-defined antigens such as diphtheria (D), tetanus (T), and more recently Haemophilus is used as one indicator of humoral immune reactivity when evaluating patients for immunodeficiency . The nature of the response to booster vaccine in this group of patients is not well defined . OBJECTIVE: To define the response to D/T booster vaccination in patients with nonprotective antibody levels in order to distinguish immunocompetent from immunodeficient children . METHODS: Patients between the ages of 16 months and 17 years referred for possible immunodeficiency were assessed for specific antibody levels as part of a standard immunologic evaluation . Twenty-six previously immunized patients had antibody titers less than or equal to 0.2 IU against D and/or T or another abnormal vaccine response . All of these patients received boosters of diphtheria and tetanus vaccine (D2T5) . Diphtheria and tetanus antibody levels were assayed 4 weeks following booster vaccination . RESULTS: Of the twenty-six subjects, a subset of patients (6) failed to show significant elevations in specific-serum antibody titers to diphtheria and/or tetanus and were thus labeled nonresponders . These patients were retrospectively compared with their responder counterparts examining specific antibody titers pre-immunization and post-immunization, serum immunoglobulins, and clinical presentation . The groups showed no significant difference in baseline specific antibody measures but following re-immunization responders showed a 31.34-fold and 22.33-fold increase in D and T antibody levels, respectively . In contrast, nonresponders produced only a 2.62-fold to D and 6.15-fold increase to T (all group comparisons P < .05) . Clinical presentation also tended to be more severe in the nonresponder group . CONCLUSIONS: These data stress the importance of specific antibody titers pre-immunization and post-immunization in the assessment of immunodeficiency states, and emphasize the different characteristics of responses between diphtheria and tetanus toxoids . The ability to achieve the minimum protective antibody level does not necessarily denote immune competence . Serum immunoglobulin levels and baseline antibody titers are insufficient for the functional assessment of the immune response . The ability to generate antibody responses following booster vaccination is a more complete measure of overall immune competence and should be considered when evaluating patients for replacement immunoglobulin therapy.

Arch Intern Med, 1997 Sep 8, 157(16), 1869 - 73
Haemophilus species bacteremia in adults . The importance of the human immunodeficiency virus epidemic; Munoz P et al.; BACKGROUND: Until the late 1970s, invasive infections caused by Haemophilus species were thought to occur mainly in children and only infrequently in adults . OBJECTIVE: To report the largest series to date of Haemophilus species bacteremia (HB) from a single center . DESIGN: Retrospective . SETTING: Large, tertiary care, general teaching hospital . METHODS: We reviewed the charts of adult patients with HB detected from January 1, 1986, to December 31, 1994 . Haemophilus strains were serotyped, and the antimicrobial resistance pattern was analyzed . RESULTS: One hundred sixteen patients had HB (0.26 cases per 1000 admissions) . Thirty-eight children and 16 adults were excluded . Human immunodeficiency virus (HIV) infection was the most common underlying condition (n = 18 {29%}), followed by malignant neoplasms (n = 12 {19%}) and chronic obstructive pulmonary disease (n = 12 {19%}) . Prevalence in HIV-positive patients was 5 cases per 1000 admissions vs 0.2 cases per 1000 admissions in HIV-negative patients . Infection was nosocomial in 16 patients (26%) . Focal diseases were pneumonia in 41 patients (66%), cholangitis in 5 patients (8%), endocarditis in 3 patients (5%), meningitis and septic arthritis each in 1 patient (2%), and primary bacteremia in 9 patients (14%) . The HIV-positive patients were significantly younger and presented more frequently with pneumonia (P < .05) . Overall, 14 patients died (22%) . Bacteremia was polymicrobial in 11 patients (18%) . Haemophilus influenzae was isolated in 53 patients (85%) . Rates of antimicrobial resistance were 11% to chloramphenicol sodium succinate, 48% to ampicillin sodium, 78% to erythromycin stearate, 76% to combined sulfamethoxazole and trimethoprim, 15% to rifampin, and 57% to clarithromycin . CONCLUSIONS: Infection with HIV has become the most common underlying disease in adults with HB in our hospital . Therapeutic approaches must take into account the high rate of antimicrobial resistance.

JAMA, 1997 Sep 3, 278(9), 705 - 11
Developing curricula to promote preventive medicine skills . The Teaching Immunization for Medical Education (TIME) Project . TIME Development Committee; Zimmerman RK et al.; CONTEXT: Vaccines are underused in the United States, resulting in needless morbidity . Many experts have concluded that clinician education is critical to increasing the nation's vaccination rates . OBJECTIVE: To develop and evaluate case-based curricular materials on immunizations that promote preventive medicine skills . DESIGN: Before-and-after trial of an educational intervention . SETTING AND PARTICIPANTS: Medical schools and primary care residency programs from 20 institutions across the United States participated in the Teaching Immunization for Medical Education (TIME) project . INTERVENTION: A multidisciplinary team developed learning objectives, abstracted clinical cases, and created case-based modules that use contextual learning and small-group interaction to solve clinical and public health problems . The case-based methods are multistation clinical teaching scenarios (MCTS) and problem-based learning (PBL) . MAIN OUTCOME MEASURES: Knowledge gained by learners from pretest to posttest and the overall ratings of the sessions by learners and facilitators based on evaluation questionnaires . RESULTS: Pretest and posttest results were obtained on a total of 1122 learners for all modules combined . For the MCTS method, mean scores increased from the 10-item pretest to the posttest by 3.1 items for measles, 3.8 for influenza, 1.8 for hepatitis B, 3.9 for pertussis, 1.9 for adult vaccination, 1.9 for childhood vaccination, and 2.6 for Haemophilus influenzae type b (P<.01 for each) . For the PBL method, mean scores increased by 3.4 items for measles, 3.3 for influenza, 2.6 for hepatitis B, and 2.5 for pertussis (P<.01 for each) . Most learners (MCTS, 98%; PBL, 89%) and most facilitators (MCTS, 97%; PBL, 100%) rated the sessions overall as very good or good . CONCLUSIONS: Use of TIME modules increases knowledge about immunizations, an essential step to improving vaccination practices of future clinicians . Given the realities of decreased faculty time and budgets, educators face major challenges in developing case-based curricula that prepare learners for the 21st century . Nationally tested libraries of cases such as the TIME modules address this dilemma.

J Bacteriol, 1997 Sep, 179(17), 5521 - 33
Mutation of the htrB gene in a virulent Salmonella typhimurium strain by intergeneric transduction: strain construction and phenotypic characterization; Sunshine MG et al.; The htrB gene product of Haemophilus influenzae contributes to the toxicity of the lipooligosaccharide . The htrB gene encodes a 2-keto-3-deoxyoctulosonic acid-dependent acyltransferase which is responsible for myristic acid substitutions at the hydroxy moiety of lipid A beta-hydroxymyristic acid . Mass spectroscopic analysis has demonstrated that lipid A from an H . influenzae htrB mutant is predominantly tetraacyl and similar in structure to lipid IV(A), which has been shown to be nontoxic in animal models . We sought to construct a Salmonella typhimurium htrB mutant in order to investigate the contribution of htrB to virulence in a well-defined murine typhoid model of animal pathogenesis . To this end, an r- m+ galE mutS recD strain of S . typhimurium was constructed (MGS-7) and used in inter- and intrastrain transduction experiments with both coliphage P1 and Salmonella phage P22 . The Escherichia coli htrB gene containing a mini-Tn10 insertion was transduced from E . coli MLK217 into S . typhimurium MGS-7 via phage P1 and subsequently via phage P22 into the virulent Salmonella strain SL1344 . All S . typhimurium transductants showed phenotypes similar to those described for the E . coli htrB mutant . Mass spectrometric analysis of the crude lipid A fraction from the lipopolysaccharide of the S . typhimurium htrB mutant strain showed that for the dominant hexaacyl form, a lauric acid moiety was lost at one position on the lipid A and a palmitic acid moiety was added at another position; for the less abundant heptaacyl species, the lauric acid was replaced with palmitoleic acid.

J Bacteriol, 1997 Sep, 179(17), 5482 - 93
TRAP transporters: a new family of periplasmic solute transport systems encoded by the dctPQM genes of Rhodobacter capsulatus and by homologs in diverse gram-negative bacteria; Forward JA et al.; The dct locus of Rhodobacter capsulatus encodes a high-affinity transport system for the C4-dicarboxylates malate, succinate, and fumarate . The nucleotide sequence of the region downstream of the previously sequenced dctP gene (encoding a periplasmic C4-dicarboxylate-binding protein) was determined . Two open reading frames (ORFs) of 681 bp (dctQ) and 1,320 bp (dctM) were identified as additional dct genes by insertional mutagenesis and complementation studies . DctQ (24,763 Da) and DctM (46,827 Da) had hydropathic profiles consistent with the presence of 4 and 12 potential transmembrane segments, respectively, and were localized in the cytoplasmic membrane fraction after heterologous expression of the dctQM ORFs in Escherichia coli . DctP, DctQ, and DctM were found to be unrelated to known transport proteins in the ABC (ATP-binding cassette) superfamily but were shown to be homologous with the products of previously unidentified ORFs in a number of gram-negative bacteria, including Bordetella pertussis, E . coli, Salmonella typhimurium, Haemophilus influenzae, and Synechocystis sp . strain PCC6803 . An additional ORF (rypA) downstream of dctM encodes a protein with sequence similarity to eukaryotic protein-tyrosine phosphatases, but interposon mutagenesis of this ORF did not result in a Dct- phenotype . Complementation of a Rhizobium meliloti dctABD deletion mutant by heterologous expression of the dctPQM genes from R . capsulatus demonstrated that no additional structural genes were required to form a functional transport system . Transport via the Dct system was vanadate insensitive, and in uncoupler titrations with intact cells, the decrease in the rate of succinate transport correlated closely with the fall in membrane potential but not with the cellular ATP concentration, implying that the proton motive force, rather than ATP hydrolysis, drives uptake . It is concluded that the R . capsulatus Dct system is a new type of periplasmic secondary transporter and that similar, hitherto-unrecognized systems are widespread in gram-negative bacteria . The name TRAP (for tripartite ATP-independent periplasmic) transporters is proposed for this new group.

J Bacteriol, 1997 Sep, 179(17), 5326 - 32
Fatty acid biosynthesis in Pseudomonas aeruginosa: cloning and characterization of the fabAB operon encoding beta-hydroxyacyl-acyl carrier protein dehydratase (FabA) and beta-ketoacyl-acyl carrier protein synthase I (FabB); Hoang TT et al.; The Pseudomonas aeruginosa fabA and fabB genes, encoding beta-hydroxyacyl-acyl carrier protein dehydratase and beta-ketoacyl-acyl carrier protein synthase I, respectively, were cloned, sequenced, and expressed in Escherichia coli . Northern analysis demonstrated that fabA and fabB are cotranscribed and most probably form a fabAB operon . The FabA and FabB proteins were similar in size and amino acid composition to their counterparts from Escherichia coli and to the putative homologs from Haemophilus influenzae . Chromosomal fabA and fabB mutants were isolated; the mutants were auxotrophic for unsaturated fatty acids . A temperature-sensitive fabA mutant was obtained by site-directed mutagenesis of a single base that induced a G101D change; this mutant grew normally at 30 degrees C but not at 42 degrees C, unless the growth medium was supplemented with oleate . By physical and genetic mapping, the fabAB genes were localized between 3.45 and 3.6 Mbp on the 5.9-Mbp chromosome, which corresponds to the 58- to 59.5-min region of the genetic map.

Infect Immun, 1997 Sep, 65(9), 3794 - 8
Heterogeneity of antibodies reactive with the dominant antigen of Actinobacillus actinomycetemcomitans; Nakashima K et al.; The serotype b-specific carbohydrate antigen (SbAg) of Actinobacillus actinomycetemcomitans Y4 is reported to be the O antigen of lipopolysaccharide, and the highest titers of serum antibody reactive with A . actinomycetemcomitans in early-onset periodontitis (EOP) patients bind SbAg . These high titers of serum antibody reactive with SbAg are associated with a lesser extent and severity of periodontal disease . The aim of this study was to determine if a limited number of genes code for anti-SbAg antibodies as has been shown for immunoglobulin G (IgG) reactive with the type b polysaccharide from Haemophilus influenzae . Serum IgG reactive with the SbAg was prepared from 20 high-titer EOP patients by affinity chromatography . The IgG subclass concentrations were determined, and heterogeneity was analyzed by isoelectric focusing (IEF) . IgG2 was the dominant subclass (83% of total IgG) in the anti-SbAg IgG fraction and represented an average of 1.33% of total serum IgG2 . The IgG2 reactive with SbAg was isolated from the affinity-purified IgG fraction by affinity chromatography with protein A and subclass-specific monoclonal antibodies . On IEF gels, only 4 to 20 bands were observed in the anti-SbAg IgG fractions, indicating limited heterogeneity . N-terminal amino acid sequence analysis of eight representative anti-SbAg IgG2 preparations indicated that variable heavy and light chains consisted largely of V(H)III and V(kappa)II, respectively . However, a significant fraction of anti-SbAg may use V(H) and V(lambda) genes with blocked N termini . In short, these findings indicate that IgG reactive with SbAg is very much like the antibody reactive with H . influenzae type b polysaccharide . Similarities include IgG2 dominance, limited bands on IEF gels, supporting an oligoclonal response, and use of genes from V(H)III and V(kappa)II regions.

Infect Immun, 1997 Sep, 65(9), 3701 - 7
Outer membrane protein D15 is conserved among Haemophilus influenzae species and may represent a universal protective antigen against invasive disease; Loosmore SM et al.; We have cloned and sequenced the d15 gene from two strains of Haemophilus influenzae type b (Hib) and two strains of nontypeable H . influenzae (NTHI) . The nucleotide and deduced protein sequences of d15 are highly conserved, with only a small variable region identified near the carboxyl terminus of the protein . Analysis of upstream sequences revealed that the H . influenzae d15 gene may be part of a large potential operon of closely spaced open reading frames, including one with significant homology to the Escherichia coli cds gene encoding CDP-diglyceride synthetase . Southern blot analysis demonstrated that the d15 gene is also present in H . influenzae types a, c, d, e, and f and in Haemophilus parainfluenzae . A recombinant D15 (rD15) protein was expressed in good quantity in E . coli from the inducible T7 promoter, and monospecific anti-rD15 antibodies were raised . Immunoblot analysis of H . influenzae serotypes a, b, c, d, e, and f, NTHI, and H . parainfluenzae lysates revealed that they all expressed a cross-reactive D15-like protein . Purified rD15 was found to be highly immunogenic in mice, guinea pigs, and rabbits, and passive transfer of anti-rD15 antibodies protected infant rats from challenge with H . influenzae type b or type a in infant rat models of bacteremia . Thus, D15 is a highly conserved antigen that is protective in animal models and it may be a useful component of a universal subunit vaccine against Haemophilus infection and disease.

Microb Pathog, 1997 Sep, 23(3), 157 - 66
Adherence of non-typeable Haemophilus influenzae promotes reorganization of the actin cytoskeleton in human or chinchilla epithelial cells in vitro; Holmes KA et al.; Non-typeable Haemophilus influenzae (NTHi) are opportunistic mucosal pathogens which adhere to epithelial cells via a variety of non-specific and specific interactions . Several adhesins have been identified and while the complimentary receptor(s) for each of these adhesins has not yet been fully characterized, it is widely accepted that adherence is an absolute prerequisite for disease . Several reports have indicated that NTHi can also be internalized and reside intracellularly . For this to occur, NTHi must be taken up by mucosal epithelial cells lining the respiratory tract . We have noted, by TEM, that adherent NTHi overlie an electron dense area in the cell membrane of human epithelial cells which is associated with a localized complex assembly of cytoskeletal fibers in the eukaryotic cytoplasm . We thus examined the potential involvement of cytoskeletal actin in this phenomenon via FITC-phalloidin labeling of respiratory tract epithelial cells which had been incubated with several clinical isolates of NTHi . Strong punctate fluorescence was coincident with adherent NTHi to both human oropharyngeal and chinchilla middle ear epithelial cells . This reactivity was similar to the discrete fluorescent spots observed with enteropathogenic Escherichia coli which were adhered to HeLa cells . In contrast, none of the NTHi isolates tested induced actin polymerization in cells of endothelial origin . While the exact mechanisms involved are yet to be elucidated, our data indicated that actin nucleation was coincident with NTHi adherence.

Wien Klin Wochenschr, 1997 Aug 8, 109(14-15), 551 - 6
Comparative genomics of mycoplasmas; Razin S; The mycoplasmas are the smallest and simplest self-replicating organisms . The goal of defining in molecular terms the entire machinery of a living cell by using mycoplasmas as models was put forward by Harold Morowitz in 1984 . The recent complete sequencing of the genomes of the human pathogens Mycoplasma genitalium and Mycoplasma pneumoniae brings us much closer to achieving this goal . The M . genitalium genome contains only 479 predicted protein coding sequences (genes) and that of M . pneumoniae 677, as compared with 1703 in Haemophilus influenzae and about 4000 in E . coli . Thus, M . genitalium is apparently the simplest organism capable of independent life with a minimal set of genes . The drastic economization in genetic information must have been associated with the parasitic mode of life of the mycoplasmas . During their reductive evolution from Gram-positive bacteria the mycoplasmas have lost the cell wall and many biosynthetic systems involved in synthesis of macromolecule building blocks provided by their host . Thus, the M . genitalium and M . pneumoniae genomes do not carry any gene involved in amino acid biosynthesis, and very few genes for vitamin, nucleic acid precursor and fatty acid biosynthesis . The mycoplasma genomes carry a minimal set of energy metabolism genes, being content with a restricted supply of ATP needed for their parasitic mode of life . Nevertheless, these minimal organisms carry the essential genes for DNA replication, transcription and translation, but even here gene saving is expressed by a minimal number of rRNA and tRNA genes . A genomic price had been paid to maintain parasitism, so that a significant number of mycoplasmal genes is devoted to adhesins, attachment organelles and variable membrane surface antigens directed towards evasion of the host immune system.

Mol Cell Probes, 1997 Aug, 11(4), 297 - 308
Rapid, sensitive, microbial detection by gene amplification using restriction endonuclease target sequences; Metherell LA et al.; The use of primers synthesized to eight class II restriction endonuclease target sequences, from Haemophilus parainfluenzae, Escherichia coli, Staphylococcus aureus, Salmonella infantis, Rhodobacter sphaeroides, Klebsiella pneumoniae, Bacillus amyloliquefaciens and Proteus vulgaris for single and multiplex PCR identification of the organisms is discussed . Results indicate that the method is sensitive and specific enough to detect single cells and attogram amounts of target DNA . It has also been demonstrated that the primers can be used in whole cell PCR for identification and whole cell PCR product recovery could be enhanced by the addition of gelatin or DMSO to PCR reaction mixtures . Other results have indicated that the method can be used for the definite identification of specific individuals present in mixed cultures or suspensions of organisms . The applicability of the method for detection of a specific strain within a group of closely related organisms has also been investigated and for that sequence/organism the results suggest that the proposed method is indeed very specific and discriminative . It is suggested that as more information becomes available regarding such sequences and their distribution, this approach could form the basis of a widescale, rapid, simple and cheap identification and/or typing system for bacteria.

Clin Exp Immunol, 1997 Aug, 109(2), 233 - 41
Haemophilus influenzae and Streptococcus pneumoniae induce different intracerebral mRNA cytokine patterns during the course of experimental bacterial meningitis; Diab A et al.; Using in situ hybridization with radiolabelled oligonucleotide probes, we studied the mRNA expression of IL-1beta, IL-4, IL-6, IL-10, IL-12, tumour necrosis factor-alpha (TNF-alpha), TNF-beta, interferon-gamma (IFN-gamma), and transforming growth factor-beta (TGF-beta) in the brain during the lethal course of experimental meningitis in a rat model inoculated intracisternally with Haemophilus influenzae type b (Hib) or Streptococcus pneumoniae and in uninfected control rats inoculated with the same volume of PBS . The production of IL-1beta, IL-4, IL-6 and IFN-gamma was also evaluated by immunohistochemistry . In the brain of Hib-inoculated rats, there was marked mRNA expression of IL-1beta, IL-6, TNF-alpha, IL-12 and IFN-gamma . IL-1beta, IL-6 and TNF-alpha were up-regulated throughout the observation period at 2, 8 and 18 h post-inoculation (p.i.), with similar patterns of induction . The Th1 cytokines IFN-gamma and TNF-beta were up-regulated within 8 h p.i . IL-10 and TGF-beta were down-regulated at 18 h p.i., while IL-4 was not detected . In contrast, the brain of S . pneumoniae-inoculated rats showed lower levels of IL-1beta, IL-6 and TNF-alpha, but higher levels of TNF-beta and detectable mRNA expression of IL-4 when compared with Hib-inoculated rats . IL-12, IFN-gamma, IL-10 and TGF-beta exhibited similar patterns of induction in the brains of Hib- and S . pneumoniae-inoculated rats . At 18 h p.i., immunohistochemistry showed similar patterns of IL-1beta, IL-4, IL-6 and IFN-gamma as mRNA expression in the brains of Hib- and S . pneumoniae-inoculated rats . The differences of cytokine profiles induced by the two bacterial strains may imply that different immunomodulating approaches should be considered, depending on etiology.

Microbiology, 1997 Aug, 143 ( Pt 8), 2783 - 95
Genomic rearrangements during evolution of the obligate intracellular parasite Rickettsia prowazekii as inferred from an analysis of 52015 bp nucleotide sequence; Andersson JO et al.; In this study a description is given of the sequence and analysis of 52 kb from the 1.1 Mb genome of Rickettsia prowazekii, a member of the alpha-Proteobacteria . An investigation was made of nucleotide frequencies and amino acid composition patterns of 41 coding sequences, distributed in 10 genomic contigs, of which 32 were found to have putative homologues in the public databases . Overall, the coding content of the individual contigs ranged from 59 to 97%, with a mean of 81% . The genes putatively identified included genes involved in the biosynthesis of nucleotides, macromolecules and cell wall structures as well as citric acid cycle component genes . In addition, a putative identification was made of a member of the regulatory response family of two-component signal transduction systems as well as a gene encoding haemolysin . For one gene, the homologue of metK, an internal stop codon was discovered within a region that is otherwise highly conserved . Comparisons with the genomic structures of Escherichia coli, Haemophilus influenzae and Bacillus subtilis have revealed several atypical gene organization patterns in the R . prowazekii genome . For example, R . prowazekii was found to have a unique arrangement of genes upstream of dnaA in a region that is highly conserved among other microbial genomes and thought to represent the origin of replication of a primordial replicon . The results presented in this paper support the hypothesis that the R . prowazekii genome is a highly derived genome and provide examples of gene order structures that are unique for the Rickettsia.

Eur Respir J, 1997 Aug, 10(8), 1754 - 60
Effect of sputum bacteriology on the quality of life of patients with bronchiectasis; Wilson CB et al.; Bronchiectatic patients have impaired health-related quality of life (QoL) and are prone to chronic lower respiratory tract infections . We have investigated whether impaired QoL is related to sputum bacteriology . Eighty seven patients with non-cystic fibrosis (non-CF) bronchiectasis, in a stable phase of their illness, completed three QoL measures, underwent a computed tomography (CT) scan and lung function tests, and provided a fresh sputum sample for microscopy and culture . The QoL of patients colonized by Pseudomonas aeruginosa (Pa group) was significantly worse than all other patients grouped together (non-Pa group), and specifically those infected by Haemophilus influenzae (Hi group) or who had no bacterial growth (NG group) (p<0.05), but not those infected by other bacterial species (O group) . The Pa group had worse lung function, but no significant differences were found between the groups for forced expiratory volume in one second (FEV1) and peak expiratory flow rate . The Pa group had significantly worse bronchiectasis scores than the O, NG and non-Pa groups, but not the Hi group . There were no significant differences between the groups with respect to the number of infective exacerbations in the last year, but the Pa group had significantly more hospital admissions . Patients infected by P . aeruginosa for more than 3 yrs had significantly worse FEV1 (p<0.03) and bronchiectasis scores (p<0.05) than those infected with P . aeruginosa for less time, but not significantly worse QoL . We conclude that, overall, patients infected with P . aeruginosa have worse quality of life, and that P . aeruginosa is associated with a greater extent of disease and worse lung function . Although patients infected with H . influenzae had extensive bronchiectasis their quality of life was better than the P . aeruginosa infected group.

Pediatr Infect Dis J, 1997 Aug, 16(8), 787 - 94
Immunogenicity and reactogenicity of a booster dose of diphtheria, tetanus, acellular pertussis and inactivated poliomyelitis vaccines given concurrently with Haemophilus type b conjugate vaccine or as pentavalent vaccine; Begue P et al.; BACKGROUND: The high reactogenicity of whole cell pertussis vaccines discourages their use in children . While acellular pertussis vaccines are less reactogenic, their use in diphtheria-tetanus-acellular pertussis (DTPa)-based combinations with Haemophilus influenzae type b conjugate vaccine (Hib) and poliomyelitis vaccines must be evaluated . OBJECTIVES: To assess the immunogenicity and reactogenicity of a booster dose of diphtheria, tetanus, acellular pertussis and inactivated poliomyelitis vaccine (DTPa-IPV) given concurrently with Hib as a mixed vaccine or as separate injections, in comparison with a whole cell pertussis vaccine (DTPw-IPV-Hib) . MATERIALS AND METHODS: This open, randomized, controlled study involved 145 healthy children ages 15 to 24 months . Immunogenicity was evaluated for all vaccine antigens, and reactogenicity was assessed with diary cards . RESULTS: Serum antibody responses to all vaccine antigens was at least as good as that observed with the pentavalent whole cell vaccine . DTPa-IPV was well-tolerated and less reactogenic than the DTPw-IPV vaccine . Administration of DTPa-IPV and Hib vaccine either separately or mixed did not alter the immunogenicity or reactogenicity profiles . CONCLUSION: DTPa-IPV vaccine, either separately or mixed with Hib vaccine, was at least as immunogenic and less reactogenic than the DTPw-Hib vaccine . Mixing DTPa-IPV and Hib vaccines did not alter the safety profile when compared with separate injections of both vaccines . A mixed DTPa-IPV-Hib vaccine can be recommended for routine use as a booster dose in primed children.

Pediatr Infect Dis J, 1997 Aug, 16(8), 780 - 7
Haemophilus influenzae type b disease and vaccination in Latin America and the Caribbean; Peltola H; OBJECTIVE: Conjugate vaccines are highly effective in preventing Haemophilus influenzae type b (Hib) diseases . Data on the epidemiology of Hib disease in Latin America and the Caribbean are not easily accessible, are incomplete and are deserving of critical analysis . METHODS: Relevant information in Spanish, English and Portuguese from over 20 regions in 16 countries was reviewed, with special attention to all Hib diseases and to children < 5 years because of their proneness to disease . Total number of cases and deaths were estimated from the incidence rates obtained from 10 studies . RESULTS: Taking into account variation between among countries, the overall incidence of Hib meningitis at age 0 to 4 years was estimated as 35 per 100,000, which would imply 20,000 cases annually in the region . An estimated rate of 60 per 100,000 for all Hib disease suggests 33,000 cases per annum . In all age groups at least 40,000 annual cases and 5000 deaths occurred annually . Given that the true role of Hib pneumonia is unknown because of data are much lacking for nonbacteremic cases, these figures are probably underestimations . Because large scale vaccination programs are being implemented in only 3 countries, the impact on Hib epidemiology has been minimal . CONCLUSIONS: Hib diseases are common in Latin America and the Caribbean, and use of conjugate vaccines is minor . Concurrently with better epidemiologic studies, their use should be extended, preferably in combination with other vaccines . Making Hib diseases notifiable would be one step for better recognition of their importance.

Ann Otol Rhinol Laryngol, 1997 Aug, 106(8), 619 - 23
Role of adenoids in the pathogenesis of otitis media: a bacteriologic and immunohistochemical analysis; Linder TE et al.; Adenoidectomy is frequently performed in children suffering from recurrent or chronic otitis media with effusion and is thought to produce a long-term effect in preventing further episodes of otitis media . Bacteriologic analysis of adenoids from 60 patients revealed a significantly elevated colonization rate of middle ear pathogens in children with a present or previous history of ear disease compared to children with adenoidal hypertrophy only . The predominant pathogen was nontypeable Haemophilus influenzae, followed by Streptococcus pneumoniae and Moraxella catarrhalis . Quantitative analysis did not demonstrate a bacterial overload in the otitis group . Lectin histochemical analysis of the adenoids revealed no significant differences between the three groups; nevertheless, colonization with S pneumoniae demonstrated an increased labeling pattern with succinylated wheat germ agglutinin, indicating the exposure of N-acetyl-glucosamine as part of its own receptor structure . On the basis of these results, we support the concept of adenoidectomy in order to remove a bacterial focus; however, we could not verify the hypothesis of bacterial overgrowth in the nasopharynx.

J Chemother, 1997 Aug, 9(4), 279 - 84
A five-day course of dirithromycin in the treatment of acute exacerbation of severe chronic obstructive pulmonary disease; Cazzola M et al.; Since dirithromycin persists at high concentrations in the lung for at least 3 days following the last dose of a 5-day course, we evaluated the clinical efficacy and tolerance of a 5-day course of dirithromycin in 20 patients with acute exacerbation of severe chronic obstructive pulmonary disease, treated with a total dose of 2.5 g dirithromycin (500 mg once-daily for 5 days) in an open, non-comparative study . Patients were assessed before therapy and after 5 (last administration), 10 (post-therapy) and 20 (late post-therapy) days . Pathogen elimination or presumed elimination was seen in 18/20 patients at the post-therapy visit and at the late post-therapy visit, but two Haemophilus influenzae out 5 were isolated in sputum after 10 days and only one after 20 days (Pseudomonas aeruginosa was the other pathogen) . Dirithromycin was well-tolerated and only 2 patients reported mild gastrointestinal pain . This study shows that a 5-day dirithromycin therapy provides a convenient and efficient dosage regimen in acute exacerbation of chronic bronchitis . Notwithstanding its poor in vitro activity against H . influenzae, dirithromycin was fairly active against this microorganism in vivo.

Clin Orthop, 1997 Aug, (341), 128 - 33
Decline of bone and joint infections attributable to haemophilus influenzae type b; Bowerman SG et al.; Haemophilus influenzae has been a major cause of infectious diseases in children and has been attributed as a significant cause of septic arthritis and osteomyelitis in children . With the advent of widespread vaccination, the incidence of Haemophilus influenzae meningitis and other infections has been well documented . This is thought to be the first report that documents the effect of vaccination on bone and joint infections . One hundred sixty-five cases of acute hematogenous osteomyelitis or septic arthritis treated at the Department of Orthopaedics at Vanderbilt University in the years before and after the advent of the Haemophilus influenzae vaccine to assess whether vaccination affected the incidence of these diseases . The data indicate that the Haemophilus influenzae vaccine has reduced to near 0 the incidence of bone and joint infections because of Haemophilus influenzae . These findings suggest that coverage of Haemophilus influenzae as part of the empiric antibiotic coverage may be no longer needed in the management of acute hematogenous osteomyelitis and septic arthritis in children.

J Bacteriol, 1997 Aug, 179(16), 5062 - 71
Characterization of a transposon Tn916-generated mutant of Haemophilus ducreyi 35000 defective in lipooligosaccharide biosynthesis; Gibson BW et al.; To define the role of the surface lipooligosaccharide (LOS) of Haemophilus ducreyi in the pathogenesis of chancroid, Tn916 mutants of H . ducreyi 35000 defective in expression of the murine monoclonal antibody (MAb) 3F11 epitope on H . ducreyi LOS were identified by immunologic screening . One mutant, designated 1381, has an LOS which lacks the MAb 3F11 epitope and migrates with an increased mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The gene disrupted by the Tn916 element in strain 1381 was identified by cloning the sequences flanking the Tn916 element . The sequences were then used to probe a lambda DASHII genomic library . In strain 1381, Tn916 interrupts a gene which encodes an open reading frame (ORF) with an Mr of 40,246 . This ORF has homology to the product of the rfaK gene of Escherichia coli . The major LOS glycoform produced by strain 1381 was analyzed by using a combination of mass spectrometry, linkage and composition analysis, and 1H nuclear magnetic resonance spectroscopy . The major LOS species was found to terminate in a single glucose attached to the heptose (L-glycero-D-manno-heptose, or Hep) trisaccharide core . In the wild-type strain 35000, glucose serves as the acceptor for the addition of the D-glycero-D-manno-heptose (or DDHep), which extends to form the mature branch of the H . ducreyi LOS . This mature oligosaccharide is in turn partially capped by the addition of sialic acid (NeuAc), i.e., NeuAc2 alpha-->3Gal beta1-->4GlcNAc beta1-->3Gal beta1-->4DDHep alpha1-->6Glc beta1 (W . Melaugh et al., Biochemistry 33:13070-13078, 1994) . Since this LOS terminates prior to the addition of the branch DD-heptose, this gene is likely to encode the D-glycero-D-manno-heptosyltransferase . Strain 1381 exhibits a significant reduction in adherence to and invasion of primary human keratinocytes . This defect was complemented by the cloned heptosyltransferase gene, indicating that the terminal portion of the LOS oligosaccharide plays an important role in adherence to human keratinocytes.

J Bacteriol, 1997 Aug, 179(16), 4977 - 84
A Salmonella typhimurium genetic locus which confers copper tolerance on copper-sensitive mutants of Escherichia coli; Gupta SD et al.; Three distinct clones from a Salmonella typhimurium genomic library were identified which suppressed the copper-sensitive (Cu(s)) phenotype of cutF mutants of Escherichia coli . One of these clones, pCUTFS2, also increased the copper tolerance of cutA, -C, and -E mutants, as well as that of a lipoprotein diacylglyceryl transferase (lgt) mutant of E . coli . Characterization of pCUTFS2 revealed that the genes responsible for suppression of copper sensitivity (scs) reside on a 4.36-kb DNA fragment located near 25.4 min on the S . typhimurium genome . Sequence analysis of this fragment revealed four open reading frames (ORF120, ORF627, ORF207, and ORF168) that were organized into two operons . One operon consisted of a single gene, scsA (ORF120), whereas the other operon contained the genes scsB (ORF627), scsC (ORF207), and scsD (ORF168) . Comparison of the deduced amino acid sequences of the predicted gene products showed that ScsB, ScsC, and ScsD have significant homology to thiol-disulfide interchange proteins (CutA2, DipZ, CycZ, and DsbD) from E . coli and Haemophilus influenzae, to an outer membrane protein (Com1) from Coxiella burnetii, and to thioredoxin and thioredoxin-like proteins, respectively . The two operons were subcloned on compatible plasmids, and complementation analyses indicated that all four proteins are required for the increased copper tolerance of E . coli mutants . In addition, the scs locus also restored lipoprotein modification in lgt mutants of E . coli . Sequence analyses of the S . typhimurium scs genes and adjacent DNAs revealed that the scs locus is flanked by genes with high homology to the cbpA (predicted curved DNA-binding protein) and agp (acid glucose phosphatase) genes of E . coli located at 22.90 min (1,062.07 kb) and 22.95 min (1,064.8 kb) of the E . coli chromosome, respectively . However, examination of the E . coli chromosome revealed that these genes are absent at this locus and no evidence has thus been obtained for the occurrence of the scs locus elsewhere on the genome.

Pediatr Nephrol, 1997 Aug, 11(4), 443 - 6
Immunization practices in children with renal disease: a report of the North American Pediatric Renal Transplant Cooperative Study; Furth SL et al.; To determine the current immunization recommendations of practicing pediatric nephrologists, a questionnaire was sent to the members of the North American Pediatric Renal Transplant Cooperative Society . Sixty-two percent of the centers responded . The results of the survey suggest that although consensus for approaching immunization does exist, recommendations do vary from center to center . Virtually all centers recommend standard vaccines {DTP, oral poliovirus (OPV), hepatitis B (Hep B), and Haemophilus influenzae B (Hib)} for their renal insufficiency and dialysis patients . Despite the fact that they are not infectious, standard killed vaccines (DTP, Hep B, Hib) are recommended less frequently for transplanted patients (86%) than their renal insufficiency (98%) and dialysis (near 100%) counterparts . Additionally, OPV and measles/mumps/rubella (MMR), both live viral vaccines, are rarely recommended post transplant . Almost 90% of centers recommend the use of influenza vaccine, while only 60% of centers recommend pneumococcal vaccine for children with renal disease . Over 70% of centers recommend the newly licensed varicella vaccine for patients on dialysis and those with renal insufficiency . Between 5% and 12% of centers recommend live viral vaccines, including OPV, MMR, and varicella vaccine, for immunosuppressed patients post renal transplant.

Antimicrob Agents Chemother, 1997 Aug, 41(8), 1770 - 4
Cloning and sequencing of a novel gene (recG) that affects the quinolone susceptibility of Staphylococcus aureus; Niga T et al.; In a study of the quinolone resistance genes in Staphylococcus aureus, a recG homolog was cloned as a gene affecting quinolone susceptibility . Sequencing analysis revealed that the gene consists of 2,061 nucleotides and encodes a 686-amino-acid polypeptide, which shows 38, 39, and 50% amino acid identity with the RecGs of Escherichia coli, Haemophilus influenzae, and Streptococcus pneumoniae, respectively . Seven helicase motifs are well conserved in the gene product . A plasmid carrying the gene complemented a recG-deficient mutant of E . coli with respect to mitomycin hypersusceptibility, demonstrating that the gene product is functionally equivalent to E . coli RecG . These results indicate that the gene is the recG gene of S . aureus . S . aureus RCM101 (recG::Tn551), designated S . aureus 3f33, is four to eight times more susceptible to quinolones than the parent strain, RCM101 . The transformation of strain 3f33 with a plasmid carrying the S . aureus recG gene made it as quinolone resistant as strain RCM101 . These results suggest that the recG gene is involved in the repair of DNA damage resulting from quinolone treatment in S . aureus.

FEMS Microbiol Lett, 1997 Aug 1, 153(1), 105 - 10
Degradation studies on Escherichia coli capsular polysaccharides by bacteriophages; Nimmich W; The serologically and structurally related Eschrichia coli capsular polysaccharides (K antigens) K13, K20, and K23 were found to be depolymerized by the bacteriophages phi K13 and phi K20 to almost similar oligomer profiles as shown by polyacrylamide gel electrophoresis . The phage-polysaccharide interactions were followed by an increase of reducing 2-keto-3-deoxyoctulosonic acid due to a phage-associated glycanase that catalyzed the hydrolytic cleavage of common beta-ketopyranosidic 2-keto-3-deoxyoctulosonic acid linkages . The related E . coli K antigens K18, K22, and K100 as well as the Haemophilus influenzae type b capsular polysaccharide were degraded by bacteriophage phi K100 with different efficacy . It is suggested that phi K100 enzymatically cleaves ribitol-5-phosphate bonds as the only structural feature present in all the polysaccharides investigated.

Microb Pathog, 1997 Aug, 23(2), 119 - 26
Kinetics of the ascension of NTHi from the nasopharynx to the middle ear coincident with adenovirus-induced compromise in the chinchilla; Miyamoto N et al.; To determine the kinetics of ascension of the eustachian tube (ET) by non-typeable Haemophilus influenzae (NTHi) in situ from the nasopharynx to the middle ear using an experimental model of otitis media (OM), we examined snap-frozen sections of chinchilla ET and middle ear mucosa for adherent bacteria over a 14 day time period . Via fluorescent- and transmission electron-microscopy, we found that NTHi preferentially adhered not to the epithelial cells but to the mucus in the ET and gradually ascended this tubal organ, reaching the middle ear approximately 10 days after intranasal inoculation of adenovirus-infected animals . The number of NTHi adherent to mucus at the pharyngeal portion of the ET increased significantly in the first 4 days after inoculation of the nares whereas the number of adherent bacteria in both the mid and tympanic portions of the ET increased more gradually over time . NTHi were not observed in the middle ear until approximately 7-10 days after inoculation of the nares which was coincident with the onset of clinical signs of OM . These data confirmed our earlier in vitro investigation which suggested that adherence to and growth within stagnant mucus within a ET compromised by adenovirus was a possible mechanism by which NTHi, resident in the nasopharynx, might gain access to the middle ear and induce OM.

Anal Biochem, 1997 Aug 1, 250(2), 228 - 36
Direct and indirect methods for molar-mass analysis of fragments of the capsular polysaccharide of Haemophilus influenzae type b; D'Ambra A et al.; Two methods are described for direct molar-mass measurement of low-molar-mass fragments obtained by oxidative cleavage of the capsular polysaccharide of Haemophilus influenzae type b . Absolute molar masses were determined by size-exclusion chromatography (SEC) with detection by multiangle laser-light-scattering photometry (MALLS) and differential refractometry (RI) . The end-group structure of the polysaccharide fragments allowed the direct measurement of average chain length by quantitative 1H NMR, from which molar masses were derived . Variation between the molar masses obtained by the two methods ranged from 5 to 7% . When molar masses estimated by indirect methods were compared to SEC-MALLS/RI data, significant deviations were observed . Analysis by SEC with secondary calibration with dextran standards gave molar masses that exceeded the SEC-MALLS/RI data by as much as 2.5-fold . Molar masses estimated by a combination of colorimetric assays varied from the SEC-MALLS/RI data by as much as 50% . These results demonstrated the applicability and superior accuracy of the direct methods of molar-mass determination of the polysaccharide fragments.

Anal Biochem, 1997 Aug 1, 250(2), 153 - 6
Identification of proteins by matrix-assisted laser desorption ionization-mass spectrometry following in-gel digestion in low-salt, nonvolatile buffer and simplified peptide recovery; Fountoulakis M et al.; Matrix-assisted laser desorption ionization-mass spectrometry is an efficient analytical method for large-scale identification of proteins separated by two-dimensional polyacrylamide gel electrophoresis . Following in-gel digestion, the salt present in the peptide extracts is usually removed by chromatography prior to analysis . Desalting is a labor-intensive and time-consuming step, limiting the total number of samples that can be processed daily . We improved the daily sample output by performing the in-gel protein digestion in low-salt, nonvolatile buffer and simplifying the recovery of the generated peptides, collecting them in a small volume by sonication . This technique is routinely used for identification of proteins of Haemophilus influenzae and human brain . The methodology described facilitates the analytical process and allows the analysis of hundreds of proteins per day . Furthermore, it represents an essential step toward process automation.

Arch Biochem Biophys, 1997 Aug 1, 344(1), 176 - 83
Purification and kinetic characterization of Haemophilus parasuis malate dehydrogenase; Wise DJ et al.; Haemophilus parasuis malate dehydrogenase ((S)-malate:NAD+ oxidoreductase; EC 1.1.1.37) isolated from cell sonicates was purified 584-fold to electrophoretic homogeneity with a 19% recovery and a specific activity of 222 units/mg protein . SDS-polyacrylamide gel electrophoresis and molecular exclusion chromatography indicated the purified enzyme to be a dimer composed of 34,600 molecular weight subunits . Kinetic parameters for all four substrates in the forward and reverse reactions indicated a sequential mechanism for this enzymic process . Product and dead-end inhibition studies were consistent with an ordered bi-bi mechanism in which NAD is the first substrate bound to the enzyme and NADH the second product released . Protection against thermodenaturation of the enzyme by NAD and not by malate was supportive of this mechanism . A pronounced product inhibition by NADH (K(i) = 9.0 microM) was observed . Although NADP did not serve as a coenzyme, a number of analogs of NAD structurally altered in the nitrogen base moieties were observed to function as coenzymes in the oxidation of malate catalyzed by the purified malate dehydrogenase . Coenzyme-competitive inhibition of the malate dehydrogenase was observed with five adenosine derivatives and six structural analogs of NAD . Of the NAD analogs studied as inhibitors, 3-pyridylcarbinol adenine dinucleotide was the most effective (K(i) = 18 microM) . Although inhibition of growth of H . parasuis by this analog was observed, it was less effective (K(i) = 136 microM) than the inhibition of the purified dehydrogenase.

J Bacteriol, 1997 Aug, 179(15), 4959 - 61
A Bacillus subtilis gene encoding a protein similar to nucleotide sugar transferases influences cell shape and viability; Price KD et al.; Bacillus subtilis gene ypfP, which is located at 196 degrees on the genetic map, shows similarity to both the monogalactosyldiacylglycerol synthase gene of Cucumis sativus, which encodes a galactosyltransferase, and the murG genes of B . subtilis, Escherichia coli, Haemophilus influenzae, and Synechocystis sp . strain PCC6803, which encode N-acetylglucosaminyltransferases involved in peptidoglycan biosynthesis . Cells containing a null mutation of ypfP are shorter and rounder than wild-type cells during growth in Luria-Bertani medium and glucose minimal medium . In addition, the mutant cells preferentially undergo lysis when grown on solid Luria-Bertani medium.

J Bacteriol, 1997 Aug, 179(15), 4949 - 52
Characterization of a periplasmic protein involved in iron utilization of Actinobacillus actinomycetemcomitans; Willemsen PT et al.; The periodontopathic bacterium Actinobacillus actinomycetemcomitans possesses a 35-kDa periplasmic iron-repressible protein . Its regulation is mediated by the Fur protein, as was inferred from the Fur-binding consensus sequence at the -35 position of the gene for the 35-kDa protein and from the relaxed expression of the gene in a mutant with an altered Fur-binding sequence . The 35-kDa protein, designated AfuA, has strong homology to HitA and FbpA of Haemophilus influenzae and Neisseria meningitidis, respectively, which serve as periplasmic iron transport proteins.

J Bacteriol, 1997 Aug, 179(15), 4937 - 41
Cloning and expression of the two genes coding for L-serine dehydratase from Peptostreptococcus asaccharolyticus: relationship of the iron-sulfur protein to both L-serine dehydratases from Escherichia coli; Hofmeister AE et al.; The structural genes sdhA and sdhB, coding for the alpha- and beta-subunits of the {4Fe-4S} cluster containing L-serine dehydratase from Peptostreptococcus asaccharolyticus, have been cloned and sequenced . Expression of modified sdhB together with sdhA in Escherichia coli led to overproduction of active His6-tagged L-serine dehydratase . E . coli MEW22, deficient in the L-serine dehydratase L-SD1, was complemented by this sdhBA construct . The derived amino acid sequence of SdhBA shares similarities with both monomeric L-serine dehydratases, L-SD1 and L-SD2, from E . coli and with a putative L-serine dehydratase from Haemophilus influenzae, which suggests that these three enzymes are also iron-sulfur proteins.

J Bacteriol, 1997 Aug, 179(15), 4815 - 20
The Haemophilus influenzae dprABC genes constitute a competence-inducible operon that requires the product of the tfoX (sxy) gene for transcriptional activation; Karudapuram S et al.; We previously showed that dprA is required for efficient processing of linear DNA during cellular transformation in Haemophilus influenzae . In this study the transcriptional regulation of dprA and two downstream genes, dprB and dprC, is examined . We demonstrate by Northern blot analysis that the dprABC genes are transcriptionally coregulated and competence inducible . We used primer extension analysis to map the transcriptional start site of dprA and of rec-2, another transformation gene involved in DNA processing . Based upon these results, we were able to identify a 26-bp dyad symmetry element immediately upstream of the -35 regions of the predicted promoters of dprA, rec-2, and two other transformation genes, comA and pilA . Finally, using transcriptional fusions of dprA to the Escherichia coli lacZ gene, we show that expression of dprA::lacZ requires tfoX and that the presence of multiple copies of tfoX abolishes the temporal regulation of dprA, resulting in its constitutive expression.

J Bacteriol, 1997 Aug, 179(15), 4768 - 77
Putative evolutionary origin of plasmids carrying the genes involved in leucine biosynthesis in Buchnera aphidicola (endosymbiont of aphids); van Ham RC et al.; An 8.5-kb plasmid encoding genes (leuABCD) involved in leucine biosynthesis and a small plasmid of 1.74 kb of yet unknown function were found in the intracellular symbiont, Buchnera aphidicola, of two divergent aphid species, Thelaxes suberi and Tetraneura caerulescens, respectively . The leuABCD-carrying plasmid (pBTs1) was amplified from total aphid DNA by inverse long PCR, using outwardly oriented oligonucleotide primers specific to leuA . The resulting 8.2-kb PCR fragment as well as the 1.74-kb plasmid (pBTc1) were cloned and sequenced . pBTs1 differed from a previously described B . aphidicola plasmid (pRPE) of the aphid Rhopalosiphum padi by the presence of a small heat shock gene (ibp) and in the order of the leuABCD and repA genes . Comparison of both leucine plasmids to the small plasmid pBTc1 revealed extensive similarity with respect to putative replication functions as well as in the presence of a highly conserved open reading frame that was found to be homologous to Escherichia coli YqhA and Haemophilus influenzae HI0507 and which may encode an integral membrane protein . The three B . aphidicola plasmids most likely evolved from a common ancestral replicon, which in turn may be distantly related to IncFII plasmids . Phylogenetic affiliations of the B . aphidicola strains of the two aphid species were assessed by sequencing of their 16S rRNA genes . Evaluation of the distribution of the leuABCD-encoding plasmids within a phylogenetic framework suggests independent origins for pBTs1 and pRPE from an ancestral replicon resembling pBTc1 . The implications for symbiotic essential amino acid biosynthesis and provisioning are discussed.

J Infect Dis, 1997 Aug, 176(2), 445 - 55
Infant immunization with pneumococcal CRM197 vaccines: effect of saccharide size on immunogenicity and interactions with simultaneously administered vaccines; Daum RS et al.; Six pentavalent pneumococcal conjugate vaccines (Pn-CRM197) were evaluated among 400 infants . The vaccines differed in saccharide chain length (oligosaccharide {OS} or polysaccharide {PS}) and saccharide quantity (0.5, 2, or 5 microg) . Subjects were randomized into groups 1-6 (Pn-CRM197 recipients) or 7 (controls) for immunization at 2, 4, and 6 months of age . Pn-CRM197 were well tolerated and elicited mean antibody concentrations that exceeded those in controls for all 5 capsular serotypes . PS formulations were generally more immunogenic than their OS counterparts . For PS vaccines, a dose-response was documented (5 microg > 2 microg > 0.5 microg), but the differences between the 5- and 2-microg formulations were insignificant . The mean anti-PRP antibody concentration was significantly higher among Pn-CRM197 recipients . It is concluded that PS vaccines are more immunogenic than OS vaccines . The improved immunogenicity from Haemophilus type b oligosaccharide conjugate (HbOC) vaccine when given with Pn-CRM197 suggests that a decreased dose of HbOC vaccine may be sufficient to elicit protection.

Infect Immun, 1997 Aug, 65(8), 3496 - 9
Purification and identification of Haemophilus ducreyi cytotoxin by use of a neutralizing monoclonal antibody; Purven M et al.; Haemophilus ducreyi produces a cytotoxin responsible for the killing of cultured human epithelial cells . Cytotoxin-neutralizing antibodies were detected in the majority of sera from patients with culture-proven chancroid, and a significantly higher level of such antibodies in patients than in blood donors was noted both in areas where the disease is endemic and those where it is not . We produced neutralizing monoclonal antibodies (MAbs) in mice with a crude osmotic preparation of the cytotoxin . These antibodies, with high capacity to neutralize cytotoxicity, were used for purification and identification of the cytotoxin . Purification was performed by a two-step procedure which included Sephacryl S-200 filtration followed by immunoaffinity chromatography . The purification resulted in poor cytotoxin protein recovery and contamination with MAbs from the affinity column . The results of the gel filtration experiments and immunoblotting indicate that the active cytotoxin consists of a single, small protein with an approximate molecular mass of 20 kDa . Cytotoxins from different strains seem to have the same or similar epitopes . The cytotoxin protein was not detected in preparations from nontoxic strains . The N-terminal amino acid sequence of the 20-kDa band was E-S-N-P-D-P-T-T-Y-P-D-V-E-L-S-P-P-P . This sequence does not resemble that of any currently known bacterial protein.

Infect Immun, 1997 Aug, 65(8), 3300 - 3
Potential role of autoantibodies in the regulation of cytokine responses during bacterial infections; Bakhiet M et al.; An immunoregulatory mechanism involving release of neutralizing autoantibodies (Aabs) to self cytokines during bacterial infections is presented herein . Intraperitoneal inoculation of Haemophilus influenzae type b into Sprague-Dawley rats resulted in a self-limiting meningitis . High levels of cells expressing mRNA for gamma interferon (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) were detected 12 to 48 h postinoculation (p.i.) in splenocytes, and large numbers of IFN-gamma-secreting cells were present in the spleen on day 3 p.i . These levels were undetectable at days 9 and 14 p.i . Increased titers of Aabs of immunoglobulin G (IgG) isotypes to both cytokines were observed, with a peak at day 7 p.i . and with very low levels at day 30 . Upon reinoculation with H . influenzae type b at day 30, regeneration of Aabs was recorded 7 days later (i.e., at day 37) . To elucidate their regulatory importance, Aabs dose-dependently inhibited IFN-gamma production by splenocytes, IFN-gamma-induced major histocompatibility complex expression by peritoneal macrophages, and TNF-alpha-induced thymocyte proliferation . To control the specificity of these Aabs, Fab fragments of purified serum Igs from day p.i . exhibited binding and neutralizing effects . Furthermore, preincubation of the sera with a cytokine inhibited the binding and neutralization effects of that particular cytokine, but not those of any other cytokine . Aab-producing B cells were cloned, and their supernatants had similar effects . Our data suggest a role for autoimmunity in cytokine regulation and suggest that a maintained balance of this mechanism may protect from sequelae.

Infect Immun, 1997 Aug, 65(8), 3267 - 70
The baboon as a nonhuman primate model for assessing the effects of maternal immunization with Haemophilus influenzae type b polysaccharide vaccines; Shearer MH et al.; These studies were performed to assess the utility of the baboon as a nonhuman primate model to evaluate vaccines for use in humans . Specifically, we examined the antibody response of baboons immunized during the third trimester of pregnancy with Haemophilus influenzae type b (Hib) polyribosylribitol phosphate (PRP) conjugate and unconjugated polysaccharide vaccines . Some of the vaccinated mothers failed to respond to a single immunization with unconjugated Hib PRP . Specific Hib PRP immunoglobulin G (IgG) but not IgM antibodies cross the baboon placenta and are detected in the offspring . Higher-titer baboon anti-Hib PRP did not express two previously defined cross-reactive human anti-Hib PRP idiotypes and was biased towards lambda light-chain expression . Spectrotype analysis indicated that baboon anti-Hib PRP was restricted in heterogeneity and oligoclonal.

Infect Immun, 1997 Aug, 65(8), 3151 - 8
Structurally defined epitopes of Haemophilus ducreyi lipooligosaccharides recognized by monoclonal antibodies; Ahmed HJ et al.; By use of enzyme-linked immunosorbent assay and immunoblotting techniques, the migration patterns and binding epitopes of lipooligosaccharides (LOS) from 10 Haemophilus ducreyi strains were investigated with two monoclonal antibodies (MAbs), MAHD6 and MAHD7, raised against LOS from H . ducreyi ITM 2665 . Closely related LOS, with defined structures, from Haemophilus influenzae, Bordetella pertussis, Aeromonas spp., and synthetic glycoproteins were also included in the analyses . The MAbs bound to conserved epitopes of LOS exposed on the surface of H . ducreyi . The MAb MAHD6 reacted with 8 of the 10 LOS from H . ducreyi but with none of the other Haemophilus or Bordetella spp . with structurally defined LOS . It is suggested that MAb MAHD6 binds to a LOS epitope (-DD-Hepp-1-->6-beta-D-Glcp-) . This LOS epitope is not present in the hexasaccharide structure of LOS from H . ducreyi ITM 4747 (E . K . H . Schweda, A . C . Sundstrom, L . M . Eriksson, J . A . Jonasson, and A . A . Lindberg, J . Biol . Chem . 269:12040-12048, 1994) . Because MAb MAHD6 reacts with the epitope mentioned above, it also discriminates between the two LOS structures, the hexasaccharide group and the nonasaccharide group, of H . ducreyi strains . MAb MAHD7 recognizes the common conserved inner core region of the LOS because it reacts with all H . ducreyi strains and with LOS with minor components in the inner core epitope structure . Rabbit polyclonal sera raised against the LOS from strains CCUG 4438 and CCUG 7470 were tested with the 10 LOS from the H . ducreyi strains . The antiserum to CCUG 7470 reacted with all H . ducreyi strains as did MAb MAHD7, whereas the antiserum to CCUG 4438 reacted with only its homologous strain and strain ITM 4747 . Also, the LOSs of our reference strains CCUG 4438 and CCUG 7470 were structurally analyzed by use of sugar analyses and electrospray ionization-mass spectrometry . The hexasaccharide and nonasaccharide structures obtained from LOS of strains CCUG 4438 and CCUG 7470 were identical to the described LOS structures from H . ducreyi ITM 4747 and ITM 2665, respectively . In conclusion, the MAb MAHD6 recognizes an epitope present in the nonasaccharide LOS group, whereas the MAb MAHD7 recognizes a conserved epitope on LOS of H . ducreyi, which is present in all strains of H . ducreyi tested . Two major groups of oligosaccharides were distinguished by their LOS structures and the reactivity of monoclonal as well as polyclonal antibodies . The majority of H . ducreyi strains possess a nonasaccharide structure of LOS.

N Z Med J, 1997 Jul 25, 110(1048), 270 - 2
Vaccine adverse events reported in New Zealand 1990-5; Mansoor O et al.; AIM: New Zealand monitors vaccine safety through vaccinator reports of adverse events following immunisation . The rate of reporting for the commonly used vaccines during 1990-5 are presented . During this time new vaccines were added to the immunisation schedule, enabling comparison of reporting rates . METHOD: The number of events were obtained from the CARM database and a rate calculated based on estimated vaccine use . RESULTS: Injection site reaction with adult tetanus-diphtheria vaccine was most commonly reported (68/100,000); a rate five times higher than with tetanus vaccine . There were also more reported reactions from diphtheria-tetanus-pertussis-Haemophilus influenzae type b (DTPH) than DTP vaccine, with 'abnormal crying' the commonest reaction reported after 29/100,000 doses of DTPH and 3/100,000 doses of DTP . The next commonest reaction for DTPH was injection site reaction (25/100,000) which compares to 17/100,000 doses of DTP . For the other inactivated vaccines, more reports were made for fever following Hib (16/100,000), than for hepatitis B (2/100,000) or influenza (1/100,000) . The most common reports following measles-mumps-rubella (MMR) vaccine were rash (17/100,000), fever (12/100,000), and injection site inflammation (5/100,000) . There were very few reports following polio vaccine, with rash, fever and headache all reported at less than 1/100,000 doses . CONCLUSION: Although only a proportion of events are reported, the picture presented here confirms the overall safety of vaccines and the value of the adverse event monitoring system . Monitoring vaccine adverse events is an essential part of the immunisation programme.

MMWR Morb Mortal Wkly Rep, 1997 Jul 25, 46(29), 665 - 71
Status report on the Childhood Immunization Initiative: reported cases of selected vaccine-preventable diseases--United States, 1996; Neuraminidase (sialidase) activity of Haemophilus parasuis; Department of Veterinary Pathobiology MC-002, College of Veterinary Medicine, University of Illinois, Urbana 61802, USA . clichten@uiuc.edu

Neuraminidase (sialidase), a potential virulence factor in bacteria, was demonstrated in Haemophilus parasuis, an invasive swine pathogen, but not in four other pathogens of the Pasteurellaceae family: H . influenzae, H . somnus, H . paragallinarum, or Actinobacillus pleuropneumoniae . H . parasuis neuraminidase had an acidic pH optimum and a specificity for several substrates also cleaved by other bacterial neuraminidases . Similar to the neuraminidase of Pasteurella multocida, H . parasuis neuraminidase was cell associated and did not require divalent cations for activity . Exogenous sialic acid added to growth medium of H . parasuis was cleared after a lag of about 10 h and these cultures grew to a greater final density than cultures without added sialic acid, indicating that exogenous sialic acid is metabolized . The role of sialidase in providing nutrients to H . parasuis may be an important factor in its obligate parasitism.

J Infect, 1997 Jul, 35(1), 85 - 6
Haemophilus segnis cholecystitis: a case report and literature review; Carson HJ et al.; Haemophilus segnis is a normal commensal of the human oropharynx which is occasionally associated with appendicitis, endocarditis or pancreatic abscess . Haemophilus segnis in the gall-bladder from a 58-year-old white female was recently encountered . The patient recovered from surgery without incident . This case is reported because the gall-bladder is now another site which has become infected with this organism . In order to provide guidance to physicians when H . segnis organism is identified, microbiologists should be aware of its behaviour in different sites.

J Infect, 1997 Jul, 35(1), 41 - 6
Procalcitonin as a marker of bacterial sepsis in patients infected with HIV-1; Gerard Y et al.; Procalcitonin (ProCT) is a recently described marker of severe sepsis . It was decided to assess the value of proCT as a marker of secondary infection in patients infected with HIV-1 . ProCT plasma levels were measured by immunoluminometric assay in a prospective study in 155 HIV-infected individuals: 102 asymptomatic and 53 with lever or suspected secondary infections . The baseline plasma level of ProCT was low (0.5 ng/ml +/- 0.37), even in the latest stages of the disease, and did not differ from the values of healthy subjects (0.54 ng/ml +/- 0.08) . EDTA-treated whole blood was collected from patients before starting specific antimicrobial therapy . No elevation of ProCT level was detected in HIV-infected patients with evolving secondary infections including PCP (n = 4), cerebral toxoplasmosis (n = 4), viral infections (n = 9), mycobacterial infections (n = 5), localized bacterial (n = 12) and fungal infections (n = 4), malignancies (n = 3), and in various associated infectious and non-infectious febrile events (n = 13) . All these plasma values were lower than 2.1 ng/ml . In contrast, high ProCT plasma levels were detected in one HIV-infected patient with a septicaemic Haemophilus influenzae infection (16.5 ng/ml) and another one with a septicaemic Pseudomonas aeruginosa infection (44.1 ng/ ml), ProCT values decreased rapidly under appropriate therapy . ProCT seems to be a specific marker of bacterial sepsis in HIV-infected patients, as no increase in other secondary infections could be detected in those patients . A rapid determination of ProCT level could be useful to confirm or refute bacterial sepsis for a better management of febrile HIV-infected patients.

Kekkaku, 1997 Jul, 72(7), 435 - 42
{Epidemiological evaluation of mycobacteria isolates in one city hospital: reports from the hospital microbiology laboratory}; Tazawa S et al.; The frequency of mycobacteria isolated from patient's specimens at Showa University Fujigaoka Hospital was investigated . By fitting a polynominal curve (degree = 3) of the annual frequency of culture-positive Mycobacterium tuberculosis (1977 through 1995), it was noted that the frequency had not changed since 1977 . The patients in the 40s or older and 60s or older comprised 74 and 38%, respectively . Of 104 patients diagnosed as tuberculosis (between 1993 and 1995), 43 (41%) were compromised hosts with the following underlying diseases: kidney disease; diabetes mellitus; malignant tumor; respiratory disease; Behcet's disease; ophthalmosarcoidosis; multiple arthritis; Hashimoto's disease . This suggested that these compromised hosts are at high risk of onset and relapse of tuberculosis, and occasionally the doctor's or patient's delay was seen during the diagnostic process . By fitting a polynominal curve (degree = 3) of the annual frequency of culture-positive atypical mycobacteria (1977 through 1995), it was noted that the frequency had increased since 1981 . The patients in the 40s or older and 60s or older comprised 88 and 60%, respectively . Between 1982 and 1994, we encountered 46 cases of atypical mycobacteriosis of the lung: 37 M . avium complex (MAX) diseases; 7 M . kansasii diseases; one M . chelonae disease; one unidentified disease involving Runyon Group II mycobacterium . Eight involved patients with bronchiectasia (5 cases), diabetes mellitus (2 cases), or leukaemia (one case) . Haemophilus influenzae, Pseudomonas aeruginosa, and Moraxella catarrhalis at more than 10(7) CFU per ml of sputum were isolated from 6 patients diagnosed with MAC or M . kansasii lung diseases, suggesting the possibility of mixed infections . M . tuberculosis and atypical mycobacterium (15 cases), and two different atypical mycobacteria (16 cases) were isolated from the same or different specimens of the same patients at the same or different times . However, the pathogenicity of these mycobacteria remained unknown, because atypical mycobacteria are non-pathogenic in many cases . The above findings suggested that the environment fit for the mycobacteria growth in human body has gradually been formed associating with aging, lung-lesion, and decline of immune capacity.

Am J Physiol, 1997 Jul, 273(1 Pt 2), H380 - 6
Role of leukocytes in cerebral autoregulation and hyperemia in bacterial meningitis in rabbits; Slater AJ et al.; The effect of leukocytes on regional cerebral blood flow (rCBF) and cerebrovascular autoregulation in experimental meningitis was determined in rabbits . Four groups of animals were studied . Cerebrospinal fluid (CSF) leukocyte migration was prevented in two groups by pretreatment with 1.5 mg/kg of IB4, a monoclonal antibody directed against CD11/18 leukocyte adhesion receptors . Intracisternal inoculation was performed with saline (control and control-IB4 groups) or Haemophilus influenzae type b (Hib and Hib-IB4 groups) . Eighteen hours later, rCBF was determined with radiolabeled microspheres . Autoregulation was assessed by graded hemorrhagic hypotension . Compared with untreated meningitis (Hib group), IB4-pretreated meningitis (Hib-IB4 group) was associated with a reduced CSF leukocyte count (1,980 +/- 880 vs . 200 +/- 110 cells/microliter; P < 0.05) and an elevated CSF colony count (2.87 +/- 0.08 vs . 5.63 +/- 0.72 log10colony-forming units/ml; P < 0.05) . Compared with control, baseline CBF was elevated in both untreated and IB4-pretreated meningitis (51 +/- 2, 54 +/- 2, 66 +/- 5, and 102 +/- 17 ml.100 g-1.min-1 in control, control-IB4, Hib, and Hib-IB4 groups, respectively) . The degree of hyperemia in meningitis was related to the CSF colony count, with a high CBF occurring in animals with high colony counts . During hypotension, CBF remained at or above baseline in the Hib group and both control groups, indicating preservation of cerebrovascular autoregulation in untreated Hib meningitis . In the Hib-IB4 group, the elevated baseline CBF was not maintained during hypotension, falling to 51% of baseline at a cerebral perfusion pressure of 30 mmHg and indicating impairment of cerebrovascular autoregulation . These results suggest that CSF leukocytes are not primarily responsible for the hyperemic response in Hib meningitis . Cerebral hyperemia may be induced either directly by bacterial components or indirectly by components of the inflammatory cascade that precede CSF leukocyte migration.

J Vet Diagn Invest, 1997 Jul, 9(3), 237 - 43
Immunohistochemical detection of Haemophilus parasuis serovar 5 in formalin-fixed, paraffin-embedded tissues of experimentally infected swine; Segales J et al.; An avidin-biotin complex immunohistochemistry technique was developed to detect Haemophilus parasuis serovar 5 in experimentally infected 18-21-day-old conventional pigs, using a rabbit polyclonal antiserum . Seven of 10 intratracheally inoculated animals developed a low to medium degree of fibrinous polyserositis; meninges and pleura were the most severely affected areas . Haemophilus parasuis was recovered from 9 of 10 pigs; in 2 of them H . parasuis was isolated from tracheal swabs only . Positive immunohistochemistry results, mainly observed as free bacteria or bacteria within inflammatory cell cytoplasm in the fibrinopurulent exudate, were observed in 8 of 10 animals . Cross-reactivity with Actinobacillus pleuropneumoniae was detected but not with other gram-positive and gram-negative bacteria tested . This immunohistochemistry technique seemed to be at least as sensitive as microbiologic cultures and could be useful in studies of pathogenesis and retrospective diagnosis . However, cross-reactivity with A . pleuropneumoniae means that positive immunohistochemistry results in lung tissue from field cases would be dubious.

Can J Vet Res, 1997 Jul, 61(3), 207 - 13
Bovine IgG2a antibodies to Haemophilus somnus and allotype expression; Corbeil LB et al.; Bovine IgG2a has been implicated in protection against pyogenic infections, including those caused by Haemophilus somnus . To further investigate the role of IgG2a in defense against H . somnus, IgG1 and IgG2a antibodies were purified from antiserum against an immunodominant 40 kDa outer membrane protein (p40) of H . somnus, which was previously shown to passively protect calves against H . somnus pneumonia . The passive protective capacity of anti-p40 IgG1 or IgG2a was evaluated in vivo in calves . Purified anti-p40 IgG1 or IgG2a was incubated with H . somnus for 15 min before intrabronchial inoculation of calves . Bacteria incubated with anti-p40 IgG1 or IgG2a were inoculated into one caudal lung lobe and bacteria incubated with IgG1 or IgG2a from the respective preimmunization serum were inoculated into the contralateral lobe . The volumes of pneumonia in the right and left lungs were determined 24 h later . The difference in volume of pneumonia with H . somnus preincubated in IgG1 pre- and postimmunization anti p40 was less (16 cm3, P = 0.298) than the difference in volume of pneumonia with H . somnus preincubated in IgG2a pre- and postimmunization anti p40 (30 cm3, P = 0.146) . Although the differences in lesion size between pre- and postimmunization serum were not statistically significant, the trend suggests IgG2a may be more protective than IgG1 . To examine this further, the peptide specificity of these IgG1 and IgG2a antibodies to p40 was examined . After limited proteolysis of p40, IgG2a antibodies reacted with 2 peptides not recognized by IgG1 antibodies . Other peptides were recognized by both isotypes . Since these studies suggested that IgG2a may be important in protection against infection, we then investigated some aspects of the role of the 2 IgG2a allotypes, A1 and A2 . In retrospective studies of age differences in expression of IgG2a allotypes, no heterozygotes were detected in calves of 60 d old or less, and fewer heterozygotes were detected in calves 61-120 d old than in cattle older than 270 d (P < 0.01) . In a subsequent prospective study of the time course of allotype expression, Holstein calves shown to be heterozygotes expressed the IgG2aA1 allotype early but the IgG2aA2 allotype was not usually detected until 3 to 4 mo of age . Thus, both the retrospective and the prospective studies showed age related differences in expression of the IgG2aA1 and A2 allotypes . This could have implication in protection.

Can J Vet Res, 1997 Jul, 61(3), 200 - 6
Interaction of Haemophilus parasuis with nasal and tracheal mucosa following intranasal inoculation of cesarean derived colostrum deprived (CDCD) swine; Vahle JL et al.; Twenty-three cesarean derived, colostrum deprived pigs were obtained at 5 wk of age and inoculated intranasally with either 1.4 x 10(8) colony forming units of Haemophilus parasuis or sterile phosphate buffered saline . Pigs were euthanized at 4, 8, 12, 18, 26, or 36 h post-inoculation and tissues from the oropharynx and respiratory tract were obtained for qualitative bacterial culture, immunohistochemistry for H . parasuis antigens, and light and transmission electron microscopy . Haemophilus parasuis was consistently isolated from the nasal cavity (17/17, 100%) and trachea (13/17, 76%) and rarely isolated from the lung (3/17, 18%) and blood stream (1/17, 6%) of infected pigs . Antigens of H . parasuis were sporadically detected on the nasal mucosa (6/17, 35%) and trachea (8/17, 47%) . Light microscopic lesions included submucosal and intraepithelial infiltrates of neutrophils and infrequent, patchy loss of cilia . Ultrastructural changes in nasal mucosal epithelial cells included cell protrusion, loss of cilia, and dilation of the cytocavitary network . Bacteria were infrequently identified and were either within an amorphous material at the apical surface of the cilia or were between individual cilia . These results suggest H . parasuis associates with the nasal mucosa and can induce a suppurative rhinitis with nasal mucosal epithelial cell degeneration . This process may represent an initial event in the pathogenesis of H . parasuis infection of swine.

Pediatr Infect Dis J, 1997 Jul, 16(7), 675 - 9
Inactivated poliovirus vaccine alone or sequential inactivated and oral poliovirus vaccine in two-, four- and six-month-old infants with combination Haemophilus influenzae type b/hepatitis B vaccine; Halsey NA et al.; BACKGROUND: Advisory committees have recommended the increased use of inactivated poliovirus vaccine (IPV) for children . OBJECTIVES: The purpose of this study was to assess the safety and immunogenicity of three schedules using IPV administered with diphtheria and tetanus toxoids and whole cell pertussis vaccines in a dual-chambered syringe . Children also received a combination of Haemophilus influenzae type b (Hib) and hepatitis B vaccines (COMVAX) . METHODS: All infants (n = 295) received IPV and COMVAX at 2 and 4 months of age and IPV, oral poliovirus vaccine (OPV) or both vaccines at 6 months and OPV at 15 months of age . RESULTS: After two doses of IPV 96 to 100% of infants had antibodies to poliomyelitis viruses types 1, 2 and 3, and after a third dose of vaccine (IPV or OPV) all but one child had antibodies to all three poliovirus types . After two doses of COMVAX 89 and 96% of children had protective levels of antibody to Hib and hepatitis B, respectively . CONCLUSIONS: IPV is highly immunogenic in a two-dose schedule . Administration of a third dose of IPV or a dose of OPV at 6 months of age is highly effective . Simultaneous administration of a combination H . influenzae type b/hepatitis B vaccine did not interfere with the response to IPV.

Pediatr Infect Dis J, 1997 Jul, 16(7), 662 - 7
Safety and immunogenicity of concurrent administration of measles-mumps-rubella-varicella vaccine and PedvaxHIB vaccines in healthy children twelve to eighteen months old . The MMRV Study Group; Reuman PD et al.; OBJECTIVE: To determine the safety and immunogenicity of concurrent administration of measles-mumps-rubella-varicella vaccine (MMRV) and PedvaxHIB (Haemophilus influenzae type b conjugate vaccine) vs . M-M-R II and PedvaxHIB followed by an optional dose of VARIVAX 6 weeks later . DESIGN: Healthy children, 12 to 18 months of age, were randomly assigned to two groups to receive (1) MMRV and PedvaxHIB given concurrently or (2) M-M-R II and PedvaxHIB followed by an optional dose of VARIVAX 6 weeks later . SUBJECTS: The study group included 294 healthy children, ages 12 to 18 months, with a negative history of measles, mumps, rubella and varicella . MAIN OUTCOME MEASURES: The seroconversion rate and magnitude of antibody responses when MMRV was given concurrently with PedvaxHIB compared with the antibody responses when VARIVAX was given 6 weeks after M-M-R II and PedvaxHIB . RESULTS: Healthy children, 12 to 18 months of age, who received MMRV and PedvaxHIB concurrently showed immune responses similar to those in the control group who received M-M-RII vaccine with PedvaxHIB followed by VARIVAX 6 weeks later . Antibody titers for varicella were significantly lower when MMRV was administered than when varicella vaccine was given separately (0.712-fold difference, P = 0.028) . No vaccine-related serious adverse reactions were reported, and no clinically significant differences were seen in the safety profiles of the two treatment groups . CONCLUSIONS: There were no statistically significant differences in the seroconversion rates between the two treatment groups for any of the antigens tested at 6 weeks and 1 year . Significantly lower geometric mean titers for varicella were noted in the group who received MMRV compared to VARIVAX given alone . Six-week seroconversion rates, persistence of immune responses at 1 year and the frequency of local and systemic reactions were comparable when MMRV was administered with PedvaxHIB compared with M-M-R II and PedvaxHIB followed by VARIVAX 6 weeks later.

Bone Marrow Transplant, 1997 Jul, 20(1), 33 - 8
Antibody responses to tetanus toxoid and Haemophilus influenzae type b conjugate vaccines following autologous peripheral blood stem cell transplantation (PBSCT); Chan CY et al.; Accelerated granulocyte and platelet recovery following peripheral blood stem cell transplantation (PBSCT) are well documented . We hypothesize that functional immunity may also be enhanced in PBSCT and performed a phase II trial of immunizations in patients with lymphoma undergoing autologous transplantation with peripheral blood stem cells or bone marrow . Seventeen BMT and 10 PBSCT recipients were immunized at 3, 6, 12, and 24-months post-transplantation with Haemophilus influenzae type b (HIB)-conjugate and tetanus toxoid (TT) vaccines . IgG anti-HIB and anti-TT antibody concentrations were measured and compared between the two groups . Geometric mean IgG anti-HIB antibody concentrations were significantly higher for PBSCT recipients compared to BMT recipients at 24 months post-transplantation (11.3 micrograms/ml vs 0.93 microgram/ml, P = 0.051) and following the 24 month immunization (66.2 micrograms/ml vs 1.30 micrograms/ml, P = 0.006) . Similar results were noted for IgG anti-TT antibody with significantly higher geometric mean antibody concentrations in the PBSCT group at 24 months post-transplantation (182 micrograms/ml vs 21.6 micrograms/ml, P = 0.039) . Protective levels of total anti-HIB antibody were achieved earlier in PBSCT recipients compared with those of BMT recipients . PBSCT recipients had higher antigen-specific antibody concentrations following HIB and TT immunizations . These results suggest enhanced recovery of humoral immunity in PBSCT recipients and earlier protection against HIB with immunization.

Protein Expr Purif, 1997 Jul, 10(2), 237 - 46
Expression, purification, and functional analysis of the TyrR protein of Haemophilus influenzae; Zhu Q et al.; The gene that was inferred to encode the TyrR protein of Haemophilus influenzae Rd was synthesized by polymerase chain reaction and inserted into a T7-based expression vector . Methods were developed to overexpress the TyrR protein of H . influenzae in Escherichia coli and to purify the protein on a large scale . Both in vitro and in vivo functional comparisons of the H . influenzae and E . coli TyrR proteins were carried out . The TyrR protein of H . influenzae was able to bind in vitro to an operator target upstream of the aroF-tyrA gene of E . coli . In the presence of {gamma-S}ATP, the DNA binding ability of the H . influenzae TyrR protein was drastically reduced . Despite the much shorter peptide chain length (318 amino acid residues vs 513), the TyrR protein of H . influenzae was as active in repressing the aroF promoter as the TyrR protein of E . coli . Repression by both proteins was enhanced in the presence of tyrosine; however, the transcriptional activation function associated with the TyrR protein of E . coli could not be detected when the H . influenzae TyrR protein was expressed in E . coli . By computer analysis, at least five operator targets for TyrR were identified within the genomic DNA of H . influenzae . These observations show that the assignment of function to the tyrR gene of H . influenzae was correctly made . Further studies of the H . influenzae TyrR protein in comparison to its E . coli counterpart should provide valuable mechanistic information on transcriptional regulation in this system.

Cornea, 1997 Jul, 16(4), 400 - 5
Ocular flora of patients with AIDS compared with those of HIV-negative patients; Gritz DC et al.; PURPOSE: To determine whether there are quantitative or qualitative differences in the ocular flora of patients with acquired immunodeficiency syndrome (AIDS) compared with human immunodeficiency virus (HIV)-negative patients . METHODS: Forty patients with AIDS and 42 HIV-negative controls were sex and age matched . All subjects had a detailed anterior segment examination, including Schirmer's test, rose bengal staining, and quantitative cultures of the conjunctiva and lids . Statistical evaluation of the relation between AIDS, keratoconjunctivitis sicca (KCS), and ocular flora was performed . RESULTS: No differences were observed in the types or numbers of organisms isolated from the conjunctiva or lids of patients with AIDS and HIV-negative subjects . Ocular flora was not influenced by use of systemic antibiotics, level of immunosuppression as measured by CD4 lymphocyte counts, KCS, or other ocular-surface disease . One AIDS patient was colonized by large numbers of Haemophilus influenzae OU with minimal clinical signs of inflammation or infection . CONCLUSION: There do not appear to be any differences in the ocular flora of HIV-negative patients and patients with AIDS . Presence of KCS and level of immunosuppression do not appear to affect the ocular flora in patients with AIDS.

Clin Exp Immunol, 1997 Jul, 109(1), 67 - 72
Antipolysaccharide antibodies in 450 children with otitis media; Misbah SA et al.; We have measured antibodies to pneumococcal and Haemophilus polysaccharides in a prospective study of 450 children aged 2-16 years with otitis media requiring grommets (ear tubes) . Pneumococcal antibody levels were significantly higher in the 2-6 year (P < 0.004) and 7-10 year (P < 0.04) study groups in comparison with age-matched controls . There was no difference in Haemophilus antibody levels between the study and control group children for the age groups 2-6 years and 11-16 years . Haemophilus antibody levels were significantly lower in the 7-10 year (P < 0.003) group in comparison with age-matched controls . Eighty-eight out of 450 (19.6%) children had pneumococcal antibody levels below the 25th percentile . Nineteen out of 88 (21.6%) children with pneumococcal antibody levels below the 25th centile were test immunized with 23 valent Pneumococcal polysaccharide and unconjugated Haemophilus type b capsular polysaccharide . Of these 19 children (aged 4-11 years), five mounted suboptimal responses to both polysaccharide antigens, whilst one child failed to respond to Haemophilus polysaccharide alone . There was no significant difference in the prevalence of IgG subclass deficiency between the normal responders and poor responders to immunization (P = 0.12) . We found no evidence of specific polysaccharide antibody deficiency in the vast majority of the 450 children studied . However, the significance of poor antibody responses to test immunization in a small minority of children with otitis media is unclear . Long-term follow up of these children is required to determine whether poor immunization responses herald the development of frank antibody deficiency.

Antimicrob Agents Chemother, 1997 Jul, 41(7), 1594 - 7
In vitro activity of BAY 12-8039, a novel 8-methoxyquinolone, compared to activities of six fluoroquinolones against Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis; Brueggemann AB et al.; The in vitro activity of a novel 8-methoxyquinolone, BAY 12-8039, against recent clinical isolates of Streptococcus pneumoniae (n = 404), Haemophilus influenzae (n = 330), and Moraxella catarrhalis (n = 250) was evaluated . Activity was compared to those of six other fluoroquinolones: ciprofloxacin, clinafloxacin, levofloxacin, ofloxacin, sparfloxacin and trovafloxacin . BAY 12-8039 and clinafloxacin had the highest levels of activity against S . pneumoniae, both with a MIC at which 90% of the isolates were inhibited (MIC90) of 0.06 microg/ml . Trovafloxacin and sparfloxacin were the next most active agents versus S . pneumoniae (MIC90s = 0.12 microg/ml) . No differences in activity against penicillin-susceptible, -intermediate, or -resistant strains of S . pneumoniae were noted for any of the fluoroquinolones tested . MIC90s for the seven fluoroquinolones ranged from 0.008 to 0.06 microg/ml versus H . influenzae and from 0.008 to 0.12 microg/ml for M . catarrhalis . The MICs for two strains of S . pneumoniae and one strain of H . influenzae were noted to be higher than those for the general population of organisms for all of the fluoroquinolones tested . Finally, the activity of BAY 12-8039 versus S . pneumoniae was found to be diminished when MIC determinations were performed with incubation of agar dilution plates or broth microdilution trays in 5 to 7% CO2 versus ambient air.

Antimicrob Agents Chemother, 1997 Jul, 41(7), 1579 - 83
Comparison of cefdinir and cefaclor in treatment of community-acquired pneumonia; Drehobl M et al.; Six hundred ninety patients were enrolled in a multicenter, randomized, double-blind trial comparing the efficacy and safety of cefdinir with those of cefaclor in the treatment of community-acquired pneumonia . Patients received either 10 days of treatment with cefdinir (n = 347) at 300 mg twice daily or 10 days of treatment with cefaclor (n = 343) at 500 mg three times daily . Microbiological assessments were performed on sputum specimens obtained at admission and at the two posttherapy visits, if available . Respiratory tract pathogens were isolated from 538 (78%) of 690 patient admission sputum specimens, with the predominant pathogens being Haemophilus parainfluenzae, Haemophilus influenzae, Streptococcus pneumoniae, and Staphylococcus aureus . The microbiological eradication rates at the test-of-cure visit were 92% (238 of 260 pathogens) and 93% (245 of 264 pathogens) for the evaluable patients treated with cefdinir and cefaclor, respectively . A satisfactory clinical response (cure plus improvement) was achieved in 89% (166 of 187) and 86% (160 of 186) of the evaluable patients treated with cefdinir and cefaclor, respectively . Except for the incidence of diarrhea, adverse event rates while on treatment were equivalent between the two treatment groups . Diarrhea incidence during therapy was higher for patients treated with cefdinir (13.7%) than for patients treated with cefaclor (5.3%) . These results indicate that cefdinir is effective and safe in the treatment of patients with pneumonia.

Antimicrob Agents Chemother, 1997 Jul, 41(7), 1517 - 20
Comparative effectiveness and safety of cefdinir and amoxicillin-clavulanate in treatment of acute community-acquired bacterial sinusitis . Cefdinir Sinusitis Study Group; Gwaltney JM Jr et al.; Cefdinir is an extended-spectrum oral cephalosporin that is active against pathogens commonly seen in acute community-acquired bacterial sinusitis (ACABS), including Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis . Two randomized, investigator-blind, multicenter trials (one in the United States and one in Europe) compared two dosage regimens of cefdinir (600 mg once a day for 10 days and 300 mg twice a day for 10 days) to amoxicillin-clavulanate (A-C) (500 mg three times a day for 10 days) for adult and adolescent patients with ACABS . Twelve hundred twenty-nine patients entered the U.S . study, 698 with antral puncture; 569 patients entered the European study, all with antral puncture . Clinical response (cure or improvement) was determined 7 to 14 days and 3 to 5 weeks posttherapy . Microbiologic eradication rates were determined 10 to 30 days posttherapy in a subset of patients who underwent pre- and posttherapy sinus aspirate culture . Rates of adverse events and treatment discontinuations due to adverse events were examined . Cefdinir, given once or twice daily, was as effective clinically (approximately 90% cure rate) as amoxicillin-clavulanate given three times daily in the treatment of ACABS . Microbiologic eradication rates were also similar in the three groups . The major side effect was mild diarrhea, occurring in approximately 20% of each group . Cefdinir caused fewer adverse events requiring treatment discontinuation.

J Bacteriol, 1997 Jul, 179(13), 4227 - 31
Identification of two minor subunits in the pilus of Haemophilus influenzae; McCrea KW et al.; Haemophilus influenzae type b (Hib) organisms produce pili, which mediate attachment to human cells and are multimeric structures composed of a 24-kDa subunit called pilin or HifA . Although pili from other organisms contain additional proteins accessory to pilin, no structural components other than pilin have been identified in Hib pili . Previous analysis of a Hib pilus gene cluster, however, suggested that two genes, hifD and hifE, may encode additional pilus subunits . To determine if hifD and hifE encode pilus components, the genes were overexpressed in Escherichia coli and the resulting proteins were purified and used to raise polyclonal antisera . Antisera raised against C-terminal HifD and HifE fragments reacted with H . influenzae HifD and HifE proteins, respectively, on Western immunoblots . Western immunoblot analysis of immunoprecipitated Hib pili demonstrated that HifD and HifE copurified with pili . In enzyme-linked immunosorbent assays, antisera raised against a recombinant HifE protein that contained most of the mature protein reacted more to piliated Hib than to nonpiliated Hib or to a mutant containing a hifE gene insertion . Immunoelectron microscopy confirmed that the HifE antiserum bound to pili and demonstrated that the antiserum bound predominantly to the pilus tips . These data indicate that HifD and HifE are pilus subunits . Adherence inhibition studies demonstrated that the HifE antiserum completely blocked pilus-mediated hemagglutination, suggesting that HifE mediates pilus adherence.

Arch Med Res, 1997 Summer, 28(2), 195 - 203
Bacterial resistance to antibiotics in acute respiratory infections (ARIs); Lopez-Antunano FJ et al.; In this review article, we make suggestions on how to approach the increasing problem worldwide of bacterial acute respiratory infections resistant to antibiotics . After a brief description of the main mechanisms of bacterial resistance, i.e., enzymatic inactivation by beta-lactamases, reduction in the permeability of the outer membrane and the development of PBPs that have decreased affinity for the antibiotic, we analyze documented experiences on the response to different groups of antibiotics (beta- lactam antibiotics, cephalosporins, carbapenems and quinolones), of the most commonly isolated bacteria from invasive respiratory infections (Haemophilus influenzae, Streptococcus pneumoniae and Moraxela (Branhamella) catarrhalis . Antimicrobial agent susceptibility in vivo and in vitro testing and the correlation of their results provide the basic information for the adoption of adequate policies and strategies for better use of antibiotics in bacterial respiratory infections; proper surveillance would allow to make intelligent changes in such a policy . Standardized recommendations for clinical practice on the use of antibiotics could be misleading, iatrogenic, and could complicate the resistance problem . To prevent and control the rise and spread of bacterial resistance, an interdisciplinary approach is needed.

Infect Immun, 1997 Jul, 65(7), 2700 - 6
Role of bacterial Mn-cofactored superoxide dismutase in oxidative stress responses, nasopharyngeal colonization, and sustained bacteremia caused by Haemophilus influenzae type b; D'Mello RA et al.; Haemophilus influenzae type b, a causative agent of bacterial sepsis and meningitis in young children, contains a single superoxide dismutase (SOD), a cytoplasmic MnSOD . To study the role of this enzyme, a chromosomal sodA::lacZ mutant (M-2) was constructed . M-2 had an increased sensitivity towards oxygen and the redox-active agent paraquat . A 3.4-fold increase in sodA-lacZ expression was found in M-2 grown with oxygen supply rates between 3 and 36 mmol of O2/liter/h . In similar experiments with the wild type, assaying SodA activity, a 3.1-fold increase was found . Both the wild type and M-2 grew best at the lowest oxygen supply rate tested, consistent with the notion that H . influenzae prefers a more anaerobic environment . In the infant rat model of infection, the ability of M-2 to colonize the nasopharynx was found to be impaired, but its ability to cause invasive disease was unaffected . This suggests that after invasion, the growth disadvantage imposed by a SodA- phenotype is not limiting.

Infect Immun, 1997 Jul, 65(7), 2593 - 8
Generation of targeted nonpolar gene insertions and operon fusions in Pasteurella haemolytica and creation of a strain that produces and secretes inactive leukotoxin; Fedorova ND et al.; An efficient method for targeted gene inactivation and generation of chromosomal gene fusions in Pasteurella haemolytica has been devised and used to create an lktC::cat operon fusion by allelic exchange at the leukotoxin gene cluster (lktCABD) . A copy of the lktC gene was insertionally inactivated by using a nonpolar, promoterless cat cassette and then delivered into P . haemolytica on a shuttle vector . Plasmid incompatibility was used to detect clones where double recombination events had occurred at the chromosomal locus . The insertion in lktC did not affect expression of the downstream genes, and the mutant strain secreted an antigenic proleukotoxin that was neither leukotoxic nor hemolytic . Expression of the lktC gene in trans restored the wild-type phenotype, confirming that LktC is required for activation of the proleukotoxin to the mature leukotoxin . Construction of the lktC::cat operon fusion allowed us to quantitate leukotoxin promoter activity in P . haemolytica and to demonstrate that transcription was maximal during early logarithmic growth phase but was reduced following entry into late logarithmic phase . This allelic exchange system should be useful for future genetic studies in P . haemolytica and could potentially be applied to other members of Haemophilus-Actinobacillus-Pasteurella family, where genetic manipulation is limited.

J Biol Chem, 1997 Jun 27, 272(26), 16555 - 63
A mono-functional 3-deoxy-D-manno-octulosonic acid (Kdo) transferase and a Kdo kinase in extracts of Haemophilus influenzae; White KA et al.; Lipopolysaccharide of Haemophilus influenzae contains a single 3-deoxy-D-manno-octulosonic acid (Kdo) residue, linked to the 6' position of lipid A . In Escherichia coli and related organisms, a Kdo disaccharide is attached to lipid A . In previous studies, we cloned the gene (kdtA) encoding the E . coli Kdo transferase and demonstrated that homogeneous preparations of KdtA polypeptide catalyzed the attachment of both Kdo groups to the precursor, lipid IVA . E . coli KdtA produced only traces of mono-glycosylated product . We now show that a single Kdo is transferred to lipid IVA in extracts of H . influenzae . The mono-functional Kdo transferase of H . influenzae is membrane-bound, and the reaction is dependent upon a CMP-Kdo-generating system, as in E . coli . The specific activity of Kdo transfer to lipid IVA is 0.5-1 nmol/min/mg in H . influenzae membranes . Utilizing solubilized H . influenzae membranes, milligram quantities of Kdo-lipid IVA were prepared for analysis . Matrix-assisted laser desorption/ionization mass spectrometry revealed a parent ion (M - H)- at m/z 1626.0, consistent with the addition of a single Kdo moiety . Like lipid IVA, Kdo-lipid IVA was an excellent substrate for the bi-functional Kdo transferase of E . coli . In membranes of H . influenzae, but not E . coli, Kdo-lipid IVA was further phosphorylated in the presence of ATP, yielding a mono-phosphorylated Kdo-lipid IVA with a parent ion (M - H)- at m/z 1703.9 . The identification of the mono-functional H . influenzae Kdo transferase, which is encoded by a KdtA homologue that displays 50% identity to its E . coli counterpart, should facilitate the mechanistic dissection of more complex multi-functional Kdo transferases, like those of E . coli and Chlamydia trachomatis.

Commun Dis Intell, 1997 Jun 26, 21(13), 173 - 6
The decline of Haemophilus influenzae type b disease in Australia; Herceg A; Between July 1993 and June 1996, there were 412 cases of invasive Haemophilus influenzae type b (Hib) disease reported to the Hib Case Surveillance Scheme, 71% in children under the age of five years . Meningitis was the most frequent illness reported, followed by epiglottitis, septicaemia and pneumonia . There were 18 deaths . Thirty-four cases were classified as vaccine failures . The number of vaccine failures increased over time and the total number of cases of Hib disease fell, consistent with an increase in Hib vaccine coverage . Based on an estimated vaccine coverage of 50% in April 1995, the vaccine efficacy for all vaccines in the period was estimated to be 89% . Invasive Hib is a serious illness of childhood which is being significantly reduced by the use of Hib vaccines, and has the potential to be eliminated from this country . Vaccination providers should aim to immunise all children against Hib disease on time and according to the National Health and Medical Research Council Standard Vaccination Schedule.

FEBS Lett, 1997 Jun 16, 409(3), 357 - 60
Bacterial aspartic proteinases; Hill J et al.; Regions of genomic DNA encoding putative aspartic proteinase domains were amplified by PCR from the bacterial species, Escherichia coli and Haemophilus influenzae . Expression of each of these DNA fragments resulted in the accumulation of the corresponding recombinant proteins in insoluble aggregates . Each recombinant protein was solubilised, refolded and shown to be able to cleave synthetic peptides that have been extensively used previously as substrates for aspartic proteinases of vertebrate, fungal and retroviral origin . Each activity was completely blocked by the diagnostic aspartic proteinase inhibitor, acetyl-pepstatin . This is thus the first report demonstrating unequivocally that aspartic proteinases may be present in bacteria.

Nucleic Acids Res, 1997 Jun 15, 25(12), 2430 - 9
Avoidance of palindromic words in bacterial and archaeal genomes: a close connection with restriction enzymes; Gelfand MS et al.; Short palindromic sequences (4, 5 and 6 bp palindromes) are avoided at a statistically significant level in the genomes of several bacteria, including the completely sequenced Haemophilus influenzae and Synechocystis sp . genomes and in the complete genome of the archaeon Methanococcus jannaschii . In contrast, there is only moderate avoidance of palindromes in the small genome of the bacterium Mycoplasma genitalium and no detectable avoidance in the genomes of chloroplasts and mitochondria . The sites for type II restriction-modification enzymes detected in the given species tend to be among the most avoided palindromes in a particular genome, indicating a direct connection between the avoidance of short oligonucleotide words and restriction-modification systems with the respective specificity . Palindromes corresponding to sites for restriction enzymes from other species are also avoided, albeit less significantly, suggesting that in the course of evolution bacterial DNA has been exposed to a wide spectrum of restriction enzymes, probably as the result of lateral transfer mediated by mobile genetic elements, such as plasmids and prophages . Palindromic words appear to accumulate in DNA once it becomes isolated from restriction-modification systems, as demonstrated by the case of organellar genomes . By combining these observations with protein sequence analysis, we show that the most avoided 4-palindrome and the most avoided 6-palindrome in the archaeon M.jannaschii are likely to be recognition sites for two novel restriction-modification systems.

Proc Natl Acad Sci U S A, 1997 Jun 10, 94(12), 6571 - 6
Vaccination against colonizing bacteria with multiple serotypes; Lipsitch M; Conjugate vaccines protect vaccinated individuals against both disease from and nasopharyngeal carriage of Streptococcus pneumoniae and Haemophilus influenzae . Protection is specific to the capsular serotype(s) included in the vaccine . This specificity has raised concern that vaccination against particular ("targeted") serotypes may cause an increase in carriage of (and diseases attributable to) nontargeted serotypes . I analyzed a mathematical model designed to predict the factors affecting, and the expected extent of, such replacement in the host population . The conditions for competitive exclusion and coexistence of serotypes under mass vaccination are derived, and the equilibrium carriage of target and nontarget serotypes is determined under various ecological and epidemiological conditions . The eradication threshold for a target serotype in the presence of competing, nontarget serotypes is always lower for serotype-specific than for bivalent vaccines . In a two-serotype model, the increase in the prevalence of any single nontargeted serotype due to vaccination will not exceed the total reduction in prevalence of a targeted serotype . However, if three or more serotypes interact epidemiologically, vaccination against one type may increase carriage of a second more than it decreases carriage of the first . Carriage of a second serotype against which the vaccine offers only partial protection may initially increase and then decrease as a function of vaccine coverage . I discuss the extent to which these theoretical results can account for existing data on serotype replacement after vaccination against H . influenzae and their implications for vaccine policy.

Vaccine, 1997 Jun, 15(9), 976 - 87
Effect of lipid modification on the physicochemical, structural, antigenic and immunoprotective properties of Haemophilus influenzae outer membrane protein P6; Yang YP et al.; The outer membrane lipoprotein, P6 of Haemophilus influenzae was studied to determine the importance of the native palmitoyl moiety on its physicochemical and immunological properties . A recombinant P6 (rP6) molecule devoid of lipidation signal sequence was expressed in Escherichia coli and its properties were compared to those of the palmitylated protein purified from H . influenzae . The isoelectric point of rP6 was more acidic than that of the native protein and also exhibited less secondary structure than P6 as judged by circular dichroism . However, both forms of P6 induced identical P6-specific antibody titers in guinea pigs when Freund's adjuvant was used . These antisera reacted with a panel of overlapping P6 peptides in a comparable manner and in addition, rabbit antisera raised against the P6 peptides reacted equally well with P6 and rP6 . Furthermore, all human convalescent sera tested exhibited similar anti-P6 and anti-rP6 antibody titers . However, rP6 was less immunogenic than P6 when administered either without adjuvant or in alum and when tested in competitive inhibition studies with anti-P6 antibodies, was a less effective inhibitor than native P6, suggesting a diminution in some of the antigenic activity of rP6 . In spite of these differences, rP6 was capable of eliciting a protective antibody response against live H . influenzae type b challenge in a modified infant rat model of bacteremia . These findings demonstrate that the non-fatty acylated rP6 could possibily be substituted for native P6 in a vaccine against H . influenzae.

Vaccine, 1997 Jun, 15(9), 955 - 61
Relative immunogenicity and efficacy of two synthetic chimeric peptides of fimbrin as vaccinogens against nasopharyngeal colonization by nontypeable Haemophilus influenzae in the chinchilla; Bakaletz LO et al.; The OMP P5-homologous fimbriae of nontypeable Haemophilus influenzae (NTHi) are an adhesin and a virulence factor for otitis media in chinchilla models . We synthesized two peptides (LB1 and LB2) which incorporate determinants of the fimbrial subunit co-linearly synthesized with a "promiscuous" T-cell epitope from the fusion protein of measles virus . Sera obtained from immunized rabbits and chinchillas demonstrated significant reciprocal titers against both the homologous peptide and isolated fimbrial protein . Antisera also immunolabeled native fimbriae of whole unfixed NTHi . Immunization with LB1 or fimbrin resulted in elimination of NTHi from the chinchilla nasopharynx 2-3 weeks earlier than controls, respectively.

Med Microbiol Immunol (Berl), 1997 Jun, 186(1), 63 - 73
The lppC gene of Streptococcus equisimilis encodes a lipoprotein that is homologous to the e (P4) outer membrane protein from Haemophilus influenzae; Gase K et al.; We report the cloning, sequencing, and analysis of a novel chromosomal gene of Streptococcus equisimilis strain H46A that codes for a membrane lipoprotein, designated LppC . The lppC gene is located 3' adjacent to, and co-oriented with, the unrelated gapC gene that encodes the previously characterized glyceraldehyde-3-phosphate dehydrogenase . Sequencing of lppC revealed an 855-bp open reading frame that predicted a 32.4-kDa polypeptide possessing a potential lipoprotein signal sequence and modification site (VTGC) . Signal sequence processing of LppC synthesized in the homologous host or expressed from plasmid pLPP2 in Escherichia coli was sensitive to globomycin, a selective inhibitor of lipoprotein-specific signal peptidase II . Subcellular localization of LppC using polyclonal antibodies raised to the hexahistidyl-tagged protein proved LppC to be tightly associated with the cytoplasmic membrane of S . equisimilis and with the outer membrane of E . coli JM109 (pLPP2) . Southern, Northern and Western analyses indicated that lpp was conserved in S . pyogenes, and transcribed independently of gap as monocistronic 0.9-kb mRNA from a sigma 70-like consensus promoter . Database searches found homology of LppC to the hel gene-encoded outer membrane protein e (P4) from Haemophilus influenzae to which it exhibits 58% sequence similarity . However, unlike the hel gene, lppC was unable to complement hemA mutants of E . coli for growth on hemin as sole porphyrin source in aerobic conditions . Furthermore, neither the wild type nor an lppC insertion mutant of S . equisimilis could grow on hemin in iron-limited medium . These results, together with findings indicating that S . equisimilis H46A had no absolute requirement for iron, led us to conclude that lppC, in contrast to hel, is not involved in hemin utilization and has yet to be assigned a function.

Acta Paediatr Jpn, 1997 Jun, 39(3), 317 - 21
The changing trend in the pattern of infective etiologies in childhood acute lower respiratory tract infection; Chong CY et al.; The etiologic agents causing acute lower respiratory tract infection (LRTI) in hospitalized children were compared for 1995 and 1988 . Between May 1994 to April 1995, 397 children were admitted to Tan Tock Seng Hospital for acute LRTI compared to 240 children in 1988 . The following criteria for LRTI were used: (i) age less than 12 years with a community-acquired LRTI; (ii) presence of cough or fever of less than 2 weeks' duration; and (iii) presence of tachypnea, chest retractions or pulmonary infiltrates on chest X-ray . Sputum cultures were considered suitable for culture if there were less than 25 epithelial cells per low power field . Moraxella catarrhalis was considered only if heavy growth of more than 3+ was seen . Etiological agents were found in about 70% of patients in both studies . Viruses constituted 41.3% of the etiologic agents in 1995 but constituted only 28% in 1988; 36% had a bacterial etiology in 1995 compared to 15% in 1988 . The most common bacteria in 1995 was M . catarrhalis (34.7%) followed by non-type B Haemophilus influenzae (33%) . In contrast, in 1988, Mycoplasma (33%) was the predominant organism followed by H . influenzae (17%) and M . catarrhalis (11.4%) . The increased incidence of M . catarrhalis could be due to antibiotic selection . A mixed viral-bacterial etiology was found in 12.3% of the 1995 cohort . The majority of the bacteria were positive by sputum cultures; only 4 (3.3%) had positive blood cultures . No penicillin resistance was detected in 1988; however, in 1995, penicillin resistance was found in 17% of the Streptococcus pneumoniae, 38.5% of H . influenzae and 83% of M . catarrhalis . It was also found that 30% of the S . pneumoniae were also resistant to erythromycin, and 23% were resistant to sulfamethoxaxole-trimethoprim; 5% of the H . influenzae had multiple resistance to erythromycin, sulfamethoxazole-trimethoprim and chloramphenicol . Among those patients with antibiotic resistance, 30% had received prior antibiotics of which 18% had had two or more antibiotics, frequently erythromycin or amoxycillin/ampicillin . Judicious use of antibiotics is required to check the rising trend of antibiotic resistance.

Electrophoresis, 1997 Jun, 18(7), 1193 - 202
Two-dimensional map of Haemophilus influenzae following protein enrichment by heparin chromatography; Fountoulakis M et al.; Two-dimensional gel electrophoresis separates several hundred protein molecules in one single experiment and is efficiently used to study the products expressed by different genomes . Low-copy-number gene products are invisible on a stained two-dimensional map and must be enriched such that sufficient amounts are present for visualization and identification . We investigated the enrichment of proteins of the bacterium Haemophilus influenzae by chromatography on immobilized heparin which has affinity for growth and protein biosynthesis factors . Total soluble proteins of the microorganism were fractionated on Heparin-Actigel which resulted in enrichment of approximately 160 proteins . The eluates, representing about 40% of the applied proteins, were analyzed by two-dimensional gel electrophoresis and the protein spots were characterized by amino acid composition analysis and matrix-assisted laser desorption ionization mass spectrometry . The proteins enriched by chromatography on the heparin gel were not exclusively low-copy-number gene products and they did not exclusively belong to one single class of proteins . The proteins that bound to the heparin gel are indicated in a two-dimensional protein map which includes more than 110 newly identified proteins.

Electrophoresis, 1997 Jun, 18(7), 1184 - 92
From genome to proteome: protein map of Haemophilus influenzae; Langen H et al.; High-resolution two-dimensional (2-D) polyacrylamide gel electrophoresis allows the separation of complex biological mixtures (i.e., several hundred proteins from a bacterial cell lysate) in a single experiment . In this report proteins from Haemophilus influenzae were separated by 2-D gels and analyzed by peptide mass fingerprinting and/or amino acid analysis . By comparing the peptide mass profiles and the amino acid composition with the Haemophilus influenzae database, 119 protein spots were identified . The combination of amino acid analysis and peptide mass fingerprinting is a powerful tool for a rapid and economical identification of a large number of proteins resolved by 2-D gels . Studies on gene regulation and changes of protein expression upon drug treatment require quick and serial analysis techniques to efficiently identify potential new drug targets.

J Vet Med Sci, 1997 Jun, 59(6), 451 - 5
Effects on endotoxin pathogenicity in pigs with acute septicemia of Haemophilus parasuis infection; Amano H et al.; Changes of endotoxin in plasma, and the response of the coagulation system and blood cells in septicemia of Haemophilus parasuis infection were examined by inoculation with H . parasuis in specific pathogen-free (SPF) pigs . Eight pigs were inoculated intratracheally with 10(5), 10(6) and 10(7) colony formation units (CFU) of the strain Nagasaki (serovar 5) . All pigs died 28 to 42 hr after inoculation . Haematologically, severe leukopenia occurred 24 hr post inoculation (hpi) until death . Glucose concentration decreased from 24 hpi to death . In the coagulation system, decrease of platelet counts, prolongation of prothrombin time, activated partial thromboplastin time, and increase of fibrinogen-fibrin degradation products were observed in all inoculated pigs . Endotoxin was detected in the plasma of all the inoculated pigs from 16 hpi to death, and its concentration rose dramatically just before death . H . parasuis was re-isolated from the blood of all inoculated pigs from 16 hpi to death, and also from almost all organs and body fluids of the pigs . The pigs had microthrombi in the kidney, liver and lungs, and many also had pneumonia, meningitis and serositis . H . parasuis antigen was detected in the lesions by the immunoperoxidase technique . The results indicated that disseminated intravascular coagulation (DIC) and endotoxin shock involved aggravation of clinical signs and death on the pigs induced to septicemia of H . parasuis.

Immunology, 1997 Jun, 91(2), 193 - 6
Effect of lisofylline and pentoxifylline on the bacterial-stimulated production of TNF-alpha, IL-1 beta IL-10 by human leucocytes; van Furth AM et al.; The present study concerns the effect of the xanthine derivates lisofylline (LSF) and pentoxifylline (PTX) on the production of pro-inflammatory cytokines tumour-necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) and the de-activating cytokine interleukin-10 (IL-10) by human leucocytes during stimulation with lipopolysaccharide (LPS), heat-killed Gram-negative bacteria (GNB) or Gram-positive bacteria (GPB) . The production of TNF-alpha and IL-1 beta by leucocytes stimulated with LPS, Haemophilus influenzae type b (Hib) or Streptococcus pneumoniae was inhibited by both drugs . The production of IL-10 by leucocytes stimulated with LPS and Hib was inhibited by both xanthine derivates only at 48 hr . However, incubation of leucocytes with S . pneumoniae in the presence of LSF or PTX stimulated the production of IL-10 about four- and twofold at 24 hr and 48 hr, respectively . In all instances, the extent of inhibition or enhancement of cytokine production by LSF or PTX was equal . The divergent effects of xanthine derivates on the IL-10 production indicate the existence of distinct intracellular pathways depending on whether leucocytes are stimulated by GPB or GNB.

J Antimicrob Chemother, 1997 Jun, 39(6), 797 - 802
Concentrations of trovafloxacin in bronchial mucosa, epithelial lining fluid, alveolar macrophages and serum after administration of single or multiple oral doses to patients undergoing fibre-optic bronchoscopy; Andrews JM et al.; Concentrations of trovafloxacin were measured in serum, alveolar macrophages, epithelial lining fluid and bronchial mucosa following single and multiple oral doses . Concentrations were determined using a microbiological assay method . There were 18 subjects in the single dose and nine subjects in the multiple dose groups . After single dosing, mean concentrations in serum, alveolar macrophages, epithelial lining fluid and bronchial mucosa at 6, 12 and 24 h were as follows: 6 h, 1.41 mg/L, 19.06 mg/L, 3.01 mg/L and 1.52 mg/kg; 12 h, 0.85 mg/L, 16.22 mg/L, 4.8 mg/L and 1.01 mg/kg; 24 h, 0.37 mg/L, 10.23 mg/L, 0.93 mg/L, and no measurable concentration, respectively . After multiple dosing (approximately 6 h post-dose) the corresponding concentrations were 1.47 mg/L, 34.3 mg/L, 10.21 mg/L and 1.67 mg/kg, respectively . These concentrations exceed the MIC90s for the common respiratory pathogens, Haemophilus influenzae 0.06 mg/L, Moraxella catarrhalis 0.008 mg/L and Streptococcus pneumoniae 0.12 mg/L and suggest that trovafloxacin should be efficacious in the treatment of community- and hospital-acquired respiratory infections.

J Antimicrob Chemother, 1997 Jun, 39(6), 763 - 9
Immune response to Fusobacterium nucleatum, Prevotella intermedia and other anaerobes in children with acute tonsillitis; Brook I et al.; The number of aerobic and anaerobic bacteria was determined in the saliva of 20 children with acute group A beta-haemolytic streptococcal (GABHS) pharyngo-tonsillitis, and 20 with acute non-GABHS tonsillitis . Antibody titres to four Gram-negative anaerobic bacilli that reside in the oropharynx (Fusobacterium nucleatum, Prevotella intermedia, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans) were determined in these and 20 control patients . An average of 8.8 aerobic and anaerobic isolates per patient saliva specimens were found during the acute tonsillitis stage in both groups, and 6.9 (in GABHS tonsillitis) and 5.6 (in non-GABHS tonsillitis) 5-6 weeks later . There were 10- to 1000-fold more bacteria in the acute stages of the inflammation in both GABHS and non-GABHS groups . These bacteria were Staphylococcus aureus, Haemophilus influenzae, Moraxella catarrhalis, Peptostreptococcus spp., F . nucleatum, Prevotella spp . and Porphyromonas spp . Significantly higher antibodies levels to F . nucleatum and P . intermedia were found in the second serum sample of patients with non-GABHS pharyngo-tonsillitis (P < 0.001) and GABHS tonsillitis (P < 0.05), as compared with their first sample or the levels of antibodies in controls . The increase in the number of several aerobic and anaerobic bacteria during acute tonsillitis and the increase in antibody levels to F . nucleatum and P . intermedia, known oral pathogens, may suggest a possible pathogenic role for these organisms in acute non-GABHS and GABHS tonsillitis.

Pediatr Emerg Care . 1997 Jun;13(3):221.
Uvulitis caused by anaerobic bacteria; Brook I; OBJECTIVE: I present two children with bacteremic uvulitis due to anaerobic bacteria . RESULTS: Fusobacterium nucleatum was recovered from the blood, and Haemophilus influenzae type b was recovered from a surface uvular culture of one patient . beta-Lactamase-producing Prevotella intermedia was isolated for the blood of the other patient . Both patients responded to parenteral, followed by oral, antimicrobial therapy . CONCLUSIONS: These findings illustrate the need to send blood cultures for both aerobic and anaerobic bacteria in patients with uvulitis.

Eur J Biochem, 1997 Jun 1, 246(2), 320 - 7
Comparison of the diphtheria mutant toxin, CRM197, with a Haemophilus influenzae type-b polysaccharide-CRM197 conjugate by optical spectroscopy; Crane DT et al.; A Haemophilus influenzae type-b capsular polysaccharide-CRM197 protein conjugate vaccine was compared with unconjugated CRM197 and diphtheria toxin, its parent molecule . Using CD and fluorescence spectroscopy, it has been possible to observe differences in structure and stability to pH and temperature due to the G52-->E mutation in CRM197 and the 'glycosylation' of CRM197 in the conjugate . CRM197 resembles the 'open' conformation of diphtheria toxin {Blewitt, M . G., Chung, L . A . & London, E . (1985) Biochemistry 24, 5458-5464} and the attachment of poly(ribosyl-ribitol phosphate) carbohydrate chains results in a still 'more open' state, although only a small decrease in the amount of ordered structure was observed . Fluorescence spectra of gel-filtration column fractions of the conjugate suggest that material of higher apparent molecular size is in the 'more open' conformation . Conjugated CRM197 begins unfolding at slightly lower temperatures (25-35 degrees C) than native material (> 35 degrees C) . In the conjugate, tryptophan residues are more accessible to the non-ionic fluorescence quencher acrylamide at 35 degrees C . The conformational change observed at pH4-6 for diphtheria toxin is also observed for CRM197, but in the conjugate begins at higher pH . This may result from the presence of charged oligosaccharide residues on the surface or the conjugation methods used . The consequences of these changes in conformation and solution behaviour of the carrier protein in terms of its ability to induce a protective, T-cell-dependent response to H . influenzae polysaccharide remain to be determined.

J Pediatr, 1997 Jun, 130(6), 923 - 30
Decline of pediatric admissions with Haemophilus influenzae type b in New York State, 1982 through 1993: relation to immunizations; Liptak GS et al.; OBJECTIVES: To evaluate the impact of vaccination for Haemophilus influenzae type b (Hib) on pediatric hospital admissions in New York State, and to identify risk factors in children who continue to be admitted for Hib invasive disease . METHODS: Retrospective review of hospitalizations in New York state from 1982 through 1993 and a survey of immunization records from physician offices in Monroe Country, New York . RESULTS: In 1982, 769 children were admitted to New York state hospitals for Hib-related conditions; by 1993, this had decreased to 133 . Significant declines during the study period occurred in the age-adjusted admission rates for Hib meningitis, septicemia, pneumonia, and epiglottitis, but not for arthritis and osteomyelitis . In 1993 alone, 712 admissions, 18 deaths, and 135 episodes of morbidity were avoided . Since 1991, the rates of admissions for Hib-related conditions have remained fairly constant . Minority subjects continue to be twice as likely as white subjects to be admitted for invasive Hib disease (0.44 vs 0.17/100,000) . Children living in urban Rochester also are more likely to be admitted and less likely to be completely immunized against Hib (61%) than those living in suburban areas (82%) . CONCLUSIONS: Although Hib vaccine has had a major impact on hospital admissions for Hib-related conditions, the goal of completely eliminating Hib disease will require programs targeted at groups at high risk, such as minorities and those living in cities.

Microbiology, 1997 Jun, 143 ( Pt 6), 1861 - 6
Nucleotide sequence and analysis of the phoB-rrnE-groESL region of the Bacillus subtilis chromosome; Sadaie Y et al.; A 36 kb sequence of the phoB-rrnE-groESL region of the Bacillus subtilis chromosome at around 55 degrees has been determined . The sequenced region contains 36 ORFs including the phoB and groESL genes, and the whole rrnE operon . The phoB gene is transcribed in the direction opposite to that of chromosome replication, while most ORFs, including groESL and the rrnE operon, are transcribed in the same direction . Two newly identified tRNA genes upstream of the rrnE operon were those for Arg-tRNA and Gly-tRNA . The sequenced region contains an operon consisting of genes for degradation and uptake of mannan . The rrnE operon and its downstream ORFs are well conserved among Mycoplasma genitalium, Haemophilus influenzae, Synechocystis sp . and Methanococcus jannaschii . delta H consensus sequences are present in the promoter regions of three ORFs, including groESL.

Nippon Rinsho, 1997 Jun, 55(6), 1580 - 7
{Pathogenesis of IgA nephropathy: role of outer membranes of Haemophilus parainfluenzae antigens}; Suzuki S et al.; IgA nephropathy is characterized by IgA deposits, predominantly in the glomerular mesangium and mesangial proliferative glomerulonephritis . Concerning its pathogenesis, several investigators suggest that the deposited IgA is an antibody to viral, bacterial, or dietary antigens . Such reports strengthen the possibility of a relationship between mucosal immunity and the pathogenesis of IgA nephropathy . We previously observed that Haemophilus parainfluenzae (HP) is more commonly isolated from the pharynx of patients with IgA nephropathy than from those with other diseases . We have also identified the glomerular deposition of the outer membranes of HP antigens (OMHP) and an increased serum concentration of IgA antibodies against OMHP in patients with IgA nephropathy . These findings suggest that HP has a role in the etiology of IgA nephropathy.

Clin Infect Dis, 1997 Jun, 24(6), 1087 - 94
Haemophilus endocarditis: report of 42 cases in adults and review . Haemophilus Endocarditis Study Group; Darras-Joly C et al.; To define the clinical, microbiological, and therapeutic characteristics of haemophilus endocarditis, we reviewed the charts of 42 adults with haemophilus endocarditis (native valve disease, 37; prosthetic valve disease, five) who were followed up between 1983 and 1995 in France . The mean duration of symptoms before diagnosis was 34 days . The causative Haemophilus species were as follows: H . parainfluenzae (26 adults), H . aphrophilus (9), H . paraphrophilus (4), and H . influenzae (3) . According to the Duke criteria, 38 cases of endocarditis were definitive and four were possible . Thirty-nine patients received combination antibacterial therapy and three received therapy with a beta-lactam agent alone (mean duration, 46 days) . Arterial embolism occurred in 15 patients . Cardiac surgery was indicated for 18 patients; 16 of these surgeries were performed within 3 months . Two patients died of heart failure . In conclusion, haemophilus endocarditis is rare and is mainly due to H . parainfluenzae . Although surgery is often necessary, haemophilus endocarditis has a favorable prognosis.

Pediatr Infect Dis J, 1997 Jun, 16(6), 593 - 9
Safety and immunogenicity of a bivalent Haemophilus influenzae type b/hepatitis B vaccine in healthy infants . Hib-HB Vaccine Study Group; West DJ et al.; OBJECTIVE: To assess the safety, tolerability and immunogenicity of COMVAX, a liquid, bivalent Haemophilus influenzae type b-hepatitis B vaccine, containing the polyribosylribitol phosphate (PRP)-Neisseria meningitidis outer membrane protein complex conjugate used in the Hib vaccine, PedvaxHIB, and the yeast-derived hepatitis B surface antigen (HBsAg) used in the HB vaccine, RECOMBIVAX HB . DESIGN: Eight hundred eighty-two healthy infants, approximately 2 months of age, were enrolled in an open, multicenter (n = 11) clinical trial and randomized to receive either COMVAX (7.5 micrograms of PRP/5 micrograms of HBsAg in 0.5 ml) or concurrent injections of the liquid formulation of PedvaxHIB (P) (7.5 micrograms of PRP in 0.5 ml) and RECOMBIVAX HB (R) (5 micrograms of HBsAg in 0.5 ml) at 2, 4 and 12 or 15 months of age . Safety and tolerability were monitored after each injection . The serum concentrations of anti-PRP and anti-HBs were determined at the time of each vaccination, 2 months after the second vaccination and 1 month after the third vaccination . RESULTS: COMVAX was well-tolerated and proved to be immunologically comparable with a series of concomitant P+R injections . There were no serious adverse experiences attributable to the study vaccines . The most commonly reported nonserious adverse experiences were all events prelisted on diary cards given to parents . These included generally mild and transient signs of inflammation at the injection site (pain/ soreness, erythema, swelling/induration), somnolence and irritability . Because children are at peak risk of invasive Hib disease during the first year of life, 6 months of age (2 months after the second dose of vaccine) was designated the time of primary interest with regard to the development of anti-PRP . At that time 94.8% of the infants given COMVAX had > 0.15 microgram/ml of anti-PRP and 72.4% had > 1.0 microgram/ ml, with a geometric mean concentration (GMC) of 2.5 micrograms/ml, compared with 95.2%, 76.3% and 2.8 micrograms/ml, respectively, in recipients of P+R . The third injection given at 12 or 15 months of age induced a secondary rise in antibody . The proportions with > 0.15 microgram/ml and > 1.0 microgram/ml of anti-PRP increased to 99.3 and 92.6%, respectively, and the GMC rose to 9.5 micrograms/ml among COMVAX recipients, compared with 98.9%, 92.3% and 10.2 micrograms/ml in children given concurrent injections of P+R . In contrast to Hib few infants in countries with low endemicity of HBV infection are at near term risk of exposure to virus . Consequently the anti-HBs response after the last dose of vaccine was designated the outcome of primary interest . At 13 to 16 months of age (1 month after the third dose of vaccine) 98.4% of children given COMVAX had a protective anti-HBs concentration of > or = 10 mIU/ml with a GMC of 4468 mIU/ml, compared with 100% and a GMC of 6944 mIU/ml among children given P+R . CONCLUSIONS: COMVAX is well-tolerated by healthy infants and can induce immunity against invasive Hib disease and HBV infection using only three injections compared with six injections if separate courses of monovalent PedvaxHIB and RECOMBIVAX HB are given.

J Bacteriol, 1997 Jun, 179(12), 3851 - 7
Role of the nhaC-encoded Na+/H+ antiporter of alkaliphilic Bacillus firmus OF4; Ito M et al.; Application of protoplast transformation and single- and double-crossover mutagenesis protocols to alkaliphilic Bacillus firmus OF4811M (an auxotrophic strain of B . firmus OF4) facilitated the extension of the sequence of the previously cloned nhaC gene, which encodes an Na+/H+ antiporter, and the surrounding region . The nhaC gene is part of a likely 2-gene operon encompassing nhaC and a small gene that was designated nhaS; the operon is preceded by novel direct repeats . The predicted alkaliphile NhaC, based on the extended sequence analysis, would be a membrane protein with 462 amino acid residues and 12 transmembrane segments that is highly homologous to the deduced products of homologous genes of unknown function from Bacillus subtilis and Haemophilus influenzae . The full-length version of nhaC complemented the Na+-sensitive phenotype of an antiporter-deficient mutant strain of Escherichia coli but not the alkali-sensitive growth phenotypes of Na+/H+-deficient mutants of either alkaliphilic B . firmus OF4811M or B . subtilis . Indeed, NhaC has no required role in alkaliphily, inasmuch as the nhaC deletion strain of B . firmus OF4811M, N13, grew well at pH 10.5 at Na+ concentrations equal to or greater than 10 mM . Even at lower Na+ concentrations, N13 exhibited only a modest growth defect at pH 10.5 . This was accompanied by a reduced capacity to acidify the cytoplasm relative to the medium compared to the wild-type strain or to N13 complemented by cloned nhaC . The most notable deficiency observed in N13 was its poor growth at pH 7.5 and Na+ concentrations up to 25 mM . During growth at pH 7.5, NhaC is apparently a major component of the relatively high affinity Na+/H+ antiport activity available to extrude the Na+ and to confer some initial protection in the face of a sudden upshift in external pH, i.e., before full induction of additional antiporters . Consistent with the inference that NhaC is a relatively high affinity, electrogenic Na+/H+ antiporter, N13 exhibited a defect in diffusion potential-energized efflux of 22Na+ from right-side-out membrane vesicles from cells that were preloaded with 2 mM Na+ and energized at pH 7.5 . When the experiment was conducted with vesicles loaded with 25 mM Na+, comparable efflux was observed in preparations from all the strains.

Microb Pathog, 1997 Jun, 22(6), 321 - 9
Detection of the phosphorylcholine epitope in streptococci, Haemophilus and pathogenic Neisseriae by immunoblotting; Kolberg J et al.; The phosphorylcholine (PC) determinant in Streptococcus pneumoniae is known to be linked to the cell wall polysaccharides (C-Ps) and to the lipoteichoic acid (LTA) (Forssman antigen) of the plasma membrane . Western blotting with two PC specific murine monoclonal antibodies (MAbs) designated 145,F-2 (IgM) and 147,A-1 (IgA) showed a similar ladder-like pattern for all examined strains of S . pneumoniae and Streptococcus mitis . Purified antigens run in parallel indicated that this ladder pattern is due to the PC of LTA . Unlike other techniques, Western blotting thus enables the identification of only one of the streptococcal structures carrying the PC epitope . Gram-negative organisms were also examined, and six of 11 Haemophilus influenzae strains reacted with the MAbs . For this species, unlike the streptococci, only one fast moving band was detected . Analyses by thin-layer chromatography (TLC) detected the PC epitope in lipopolysaccharide (LPS) fraction from H . influenzae . Some strains of the Neisseriaceae family were also positive by Western blotting, but TLC and immunostaining did not detect the PC determinant in LPS.

J Infect Dis, 1997 Jun, 175(6), 1440 - 5
Relationship between nasopharyngeal colonization and the development of otitis media in children . Tonawanda/Williamsville Pediatrics; Faden H et al.; Streptococcus pneumoniae, nontypeable Haemophilus influenzae, and Moraxella catarrhalis are the predominant bacteria associated with otitis media . A cohort of 306 infants was followed from birth through 12 months to determine frequency and duration of colonization and risk of acute otitis media (AOM) and otitis media with effusion (OME) . M . catarrhalis was the most common bacterium isolated . Infants colonized at < or = 3 months of age were at increased risk of AOM and OME . Early colonization with M . catarrhalis revealed the greatest risk (relative risk {RR} = 1.24), especially for OME (RR = 1.57) . There was a strong relationship between the frequency of colonization and OM (r = .37, P < .001,) for each pathogen . Although S . pneumoniae, nontypeable H . influenzae, and M . catarrhalis are part of the normal nasopharyngeal flora during infancy, an increased rate of colonization may identify a subpopulation of children that is at increased risk of OM.

J Infect Dis, 1997 Jun, 175(6), 1390 - 5
Comparison of enzyme immunoassays for antibodies to Haemophilus ducreyi in a community outbreak of chancroid in the United States; Chen CY et al.; The performance of two EIAs (adsorption EIA and lipooligosaccharide {LOS} EIA) that detect antibodies to Haemophilus ducreyi was evaluated with serum specimens obtained from 163 patients (96 with genital ulcer disease {GUD}) . Paired serum specimens (initial and follow-up) were obtained from 52 of the GUD patients . By use of initial serum specimens from 82 GUD patients whose etiologic agents for their ulcers had been identified, the adsorption EIA had a sensitivity and specificity for chancroid of 53% and 71%, while the LOS EIA had a sensitivity and specificity of 48% and 89%, respectively . Sensitivity and specificity of the adsorption EIA increased to 78% and 84%, respectively, when the results of follow-up serum specimens were used to calculate optimal performance . The proportion of patients testing positive for H . ducreyi who had anti-H . ducreyi IgG antibodies, as determined by adsorption EIA, increased with the duration of infection, thus limiting the role of EIAs in the diagnosis of chancroid.

Drugs, 1997 Jun, 53(6), 973 - 1004
Clarithromycin . A review of its efficacy in the treatment of respiratory tract infections in immunocompetent patients; Langtry HD et al.; Clarithromycin is a broad spectrum macrolide antibacterial agent active in vitro and effective in vivo against the major pathogens responsible for respiratory tract infections in immunocompetent patients . It is highly active in vitro against pathogens causing atypical pneumonia (Chlamydia pneumoniae, Mycoplasma pneumoniae and Legionella spp.) and has similar activity to other macrolides against Staphylococcus aureus . Streptococcus pyogenes, Moraxella catarrhalis and Streptococcus pneumoniae . Haemophilus influenzae is susceptible or intermediately susceptible to clarithromycin alone, but activity is enhanced when the parent drug and metabolite are combined in vitro . Absorption of clarithromycin is unaffected by food . More than half of an oral dose is systemically available as the parent drug and the active 14-hydroxy metabolite . Pharmacokinetics are nonlinear, with plasma concentrations increasing in more than proportion to the dosage . First-pass metabolism results in the rapid appearance of the active metabolite 14-hydroxy-clarithromycin in plasma . Clarithromycin and its active metabolite are found in greater concentrations in the tissues and fluids of the respiratory tract than in plasma . Dosage adjustments are required for patients with severe renal failure, but not for elderly patients or those with hepatic impairment . Drug interactions related to the cytochrome P450 system may occur with clarithromycin use . In addition to the standard immediate-release formulation for administration twice daily, a modified-release formulation of clarithromycin is now available for use once daily . In dosages of 500 to 1000 mg/day for 5 to 14 days, clarithromycin was as effective in the treatment of community-acquired upper and lower respiratory tract infections in hospital and community settings as beta-lactam agents (with or without a beta-lactamase inhibitor), cephalosporins and most other macrolides . Clarithromycin was similar in efficacy to azithromycin in comparative studies and is as effective as and better tolerated than erythromycin . Adverse events are primarily gastrointestinal in nature, but result in fewer withdrawals from therapy than are seen with erythromycin . Clarithromycin provides similar clinical and bacteriological efficacy to that seen with beta-lactam agents, cephalosporins and other macrolides . It offers a cost-saving alternative to intravenous erythromycin use in US hospitals and is available in both once-daily and twice-daily formulations . The spectrum of activity of clarithromycin against common and emerging respiratory tract pathogens may make it suitable for use in the community as empirical therapy of respiratory tract infections in both children and adults.

Mayo Clin Proc, 1997 Jun, 72(6), 532 - 42
Infective endocarditis due to unusual or fastidious microorganisms; Berbari EF et al.; Infective endocarditis due to fastidious microorganisms is commonly encountered in clinical practice . Some organisms such as fungi account for up to 15% of cases of prosthetic valve infective endocarditis, whereas organisms of the HACEK group (Haemophilus parainfluenzae, H . aphrophilus, and H . paraphrophilus, Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens, and Kingella kingae) cause 3% of community-acquired cases of infective endocarditis . Special techniques are necessary to identify these microorganisms . A history of contact with mammals or birds may suggest infection caused by Coxiella burnetii (Q fever), Brucella species, or Chlamydia psittaci . A nosocomial cluster of postsurgical infective endocarditis may be caused by Legionella species or Mycobacterium species . If risk factors that are commonly associated with fungal infections (cardiac surgical treatment, prolonged hospitalization, indwelling central venous catheters, and long-term antibiotic use) are present, fungal endocarditis is possible . Patients with endocarditis and a history of periodontal disease or dental work in whom routine blood cultures are negative might have infection due to nutritionally variant streptococci or bacteria of the HACEK group . Communication between the microbiologist and the clinician is of crucial importance for identification of these microorganisms early during the course of the infection before complications such as embolization or valvular failure occur . In this article, we review the microbiologic and clinical features of these organisms and provide recommendations for diagnosis and treatment.

Antimicrob Agents Chemother, 1997 Jun, 41(6), 1377 - 9
Pharmacodynamic properties of BAY 12-8039 on gram-positive and gram-negative organisms as demonstrated by studies of time-kill kinetics and postantibiotic effect; Boswell FJ et al.; Time-kill kinetics of BAY 12-8039 were studied at two inocula against three strains each of Bacteroides fragilis, Escherichia coli, Staphylococcus aureus, Haemophilus influenzae, and Streptococcus pyogenes . The postantibiotic effects of BAY 12-8039 were studied on three strains each of E . coli, S . aureus, H . influenzae, Streptococcus pyogenes, and Streptococcus pneumoniae . The pharmacodynamic data demonstrated that BAY 12-8039 has marked activity against gram-positive and gram-negative organisms (under both anaerobic and aerobic conditions) and anaerobes . BAY 12-8039 also exhibited a postantibiotic effect of >1 h for all strains except one E . coli strain.

J Bacteriol, 1997 Jun, 179(11), 3670 - 5
Plasmid maintenance functions of the large virulence plasmid of Shigella flexneri; Radnedge L et al.; The large virulence plasmid pMYSH6000 of Shigella flexneri contains a replicon and a plasmid maintenance stability determinant (Stb) on adjacent SalI fragments . The presence of a RepFIIA replicon on the SalI C fragment was confirmed, and the complete sequence of the adjacent SalI O fragment was determined . It shows homology to part of the transfer (tra) operon of the F plasmid . Stb stabilizes a partition-defective P1 miniplasmid in Escherichia coli . A 1.1-kb region containing a homolog of the F trbH gene was sufficient to confer stability . However, the trbH open reading frame could be interrupted without impairing stability . Deletion analysis implicated the involvement of two small open reading frames, STBORF1 and STBORF2, that fully overlap trbH in the opposite direction . These open reading frames are closely related to the vagC and vagD genes of the Salmonella dublin virulence plasmid and to open reading frame pairs in the F trbH region and in the chromosomes of Dichelobacter nodosus and Haemophilus influenzae . Stb appears to promote better-than-random distribution of plasmid copies and is a plasmid incompatibility determinant . The F homolog does not itself confer stability but exerts incompatibility against the activity of the Stb system . Stb is likely to encode either an active partition system or a postsegregational killing system . It shows little similarity to previously studied plasmid stability loci, but the genetic organization of STBORF1 and STBORF2 resembles that of postsegregational killing mechanisms.

Infect Immun, 1997 Jun, 65(6), 2491 - 6
Identification and characterization of a DNA region involved in the export of capsular polysaccharide by Actinobacillus pleuropneumoniae serotype 5a; Ward CK et al.; Actinobacillus pleuropneumoniae synthesizes a serotype-specific capsular polysaccharide that acts as a protective barrier to phagocytosis and complement-mediated killing . To begin understanding the role of A . pleuropneumoniae capsule in virulence, we sought to identify the genes involved in capsular polysaccharide export and biosynthesis . A 5.3-kb XbaI fragment of A . pleuropneumoniae serotype 5a J45 genomic DNA that hybridized with DNA probes specific for the Haemophilus influenzae type b cap export region was cloned and sequenced . This A . pleuropneumoniae DNA fragment encoded four open reading frames, designated cpxDCBA . The nucleotide and predicted amino acid sequences of cpxDCBA contained a high degree of homology to the capsule export genes of H . influenzae type b bexDCBA, Neisseria meningitidis group B ctrABCD, and, to a lesser extent, Escherichia coli K1 and K5 kpsE and kpsMT . When present in trans, the cpxDCBA gene cluster complemented kpsM::TnphoA or kpsT::TnphoA mutations, determined by enzyme immunoassay and by restored sensitivity to a K5-specific bacteriophage . A cpxCB probe hybridized to genomic DNA from all A . pleuropneumoniae serotypes tested, indicating that this DNA was conserved among serotypes . These data suggest that A . pleuropneumoniae produces a group II family capsule similar to those of related mucosal pathogens.

Infect Immun, 1997 Jun, 65(6), 2413 - 9
Alterations in levels of DnaK and GroEL result in diminished survival and adherence of stressed Haemophilus ducreyi; Parsons LM et al.; Haemophilus ducreyi is a hemin-requiring bacterium causing the genital ulcer disease chancroid . Previously we demonstrated that the heat shock protein GroEL was immunogenic and possibly highly expressed in a mammalian host . The present study was initiated to (i) determine the relative amounts of GroEL expressed by H . ducreyi during in vitro exposure to stresses and (ii) evaluate whether a high level of GroEL is directly or indirectly required for survival and adherence of stressed H . ducreyi . Using scanning densitometry of sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles, we found that H . ducreyi expressed high basal levels of GroEL, averaging fivefold greater than in Escherichia coli . These high GroEL levels increased up to twofold upon exposure of the organism to heat shock or high levels of hydrogen peroxide and during adherence to two human genital cell lines . Furthermore, when the gene for DnaK was present on a multicopy plasmid in H . ducreyi, a 1.8-fold increase in DnaK and a 2.3-fold reduction in GroEL were seen . These results suggest that DnaK serves as a negative modulator of H . ducreyi GroEL . Subsequently we found that H . ducreyi with lower GroEL had diminished ability to survive when challenged by heat and oxidative stresses . In addition, the long, parallel chains characteristic of virulent strains of H . ducreyi were absent when GroEL was lowered, so that fewer bacterial cells adhered to the human cells . These results suggest that the unusually high basal levels of GroEL are involved, either directly or indirectly, in the survival, chaining, and adherence of H . ducreyi in the presence of the combined stresses of the host environment.

Pediatrics, 1997 Jun, 99(6), 765 - 73
Vaccine Safety Datalink project: a new tool for improving vaccine safety monitoring in the United States . The Vaccine Safety Datalink Team; Chen RT et al.; OBJECTIVE: To fill the large "gaps and limitations" in current scientific knowledge of rare vaccine adverse events identified in recent reviews of the Institute of Medicine . METHODS: Computerized information on immunization, medical outcomes, and potential confounders on more than 500 000 children 0 to 6 years of age is linked annually at several health maintenance organizations to create a large cohort for multiple epidemiologic studies of vaccine safety . RESULTS: Analysis of 3 years of follow-up data shows that 549 488 doses of diphtheria-tetanus-pertussis (DTP) and 310 618 doses of measles-mumps-rubella (MMR) vaccines have been administered to children in the study cohort . Analyses for associations between vaccines and 34 medical outcomes are underway . Screening of automated data shows that seizures are associated with receipt of DTP on the same day (relative risk {RR}, 2.1; 95% confidence interval {CI}, 1.1 to 4.0) and 8 to 14 days after receipt of MMR (RR, 3.0; 95% CI, 2.1 to 4.2) . The diversity of vaccination exposures in this large cohort permits us to show that an apparent association of seizures 8 to 14 days after Haemophilus influenzae type b vaccine (RR, 1.6; 95% CI, 1.2 to 2.1) was attributable to confounding by simultaneous MMR vaccination; the association disappears with appropriate adjustment (RR, 1.0; 95% CI, 0.7 to 1.4) . CONCLUSION: Preliminary design, data collection, and analytic capability of the Vaccine Safety Datalink project has been validated by replication of previous known associations between seizures and DTP and MMR vaccines . The diversity in vaccine administration schedules permits potential disentangling of effects of simultaneous and combined vaccinations . The project provides a model of public health-managed care collaborations in addition to an excellent infrastructure for safety and other studies of vaccines.

J Clin Microbiol, 1997 Jun, 35(6), 1630 - 2
Rapid identification of Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, and Haemophilus paraphrophilus by restriction enzyme analysis of PCR-amplified 16S rRNA genes; Riggio MP et al.; Restriction enzyme analysis of PCR-amplified 16S rRNA genes was used to distinguish among clinical isolates of Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, and Haemophilus paraphrophilus which were originally identified by conventional phenotypic methods . This PCR-based method is a reliable and rapid alternative to conventional methods for identification of these bacterial species.

J Clin Microbiol, 1997 Jun, 35(6), 1517 - 20
Outbreak of amoxicillin-resistant Haemophilus influenzae type b: variable number of tandem repeats as novel molecular markers; van Belkum A et al.; An outbreak caused by amoxicillin-resistant Haemophilus influenzae type b was noted among patients suffering from chronic obstructive pulmonary disease . Since infections were clustered in time and place, an ongoing outbreak was suspected . The spread of the strain and the course of the outbreak could be followed by random amplification of polymorphic DNA (RAPD) analysis of the different bacterial isolates . In addition, studies were aimed at the determination of length polymorphism in regions of repetitive DNA . By PCR-mediated amplification of variable number of tandem repeat regions (VNTRs), additional insight into the genome composition of the epidemic strain was gained . Our results show that VNTRs comprising repeat units that are 3, 5, or 6 nucleotides in length provided stable genetic markers that can be used for molecular typing of H . influenzae type b . VNTRs built from tetranucleotide units, however, appear to be hypervariable and not suited for epidemiological studies . The observed variability in this latter class of VNTRs might be reminiscent of the bacterium's capacity to deal with unfavorable host factors.

J Clin Microbiol, 1997 Jun, 35(6), 1311 - 5
Variability in susceptibilities of Haemophilus influenzae to clarithromycin and azithromycin due to medium pH; Nilius AM et al.; The National Committee for Clinical Laboratory Standards (NCCLS) methods for susceptibility testing of Haemophilus influenzae in Haemophilus test medium allow a pH range of 7.2 to 7.4 . However, it is known that bacteria may appear to be less susceptible to macrolides at lower pHs . Forty-four strains of H . influenzae were tested for their susceptibilities to clarithromycin and azithromycin by the disk diffusion and broth microdilution methods . The isolates appeared to be less susceptible at pH 7.2 than at pH 7.4 by both methods . Clarithromycin was less active at pH 7.2 against 43% of the isolates by the disk diffusion method and against 52% of the isolates by the broth microdilution method . Similarly, azithromycin was less active at pH 7.2 against 41 and 45% of the isolates by the disk diffusion and broth microdilution methods, respectively . Forty-two isolates were classified as clarithromycin susceptible and all isolates were classified as azithromycin susceptible by the disk diffusion method, regardless of the medium pH . However, only 21 isolates were clarithromycin susceptible at pH 7.2 and 34 isolates were susceptible at pH 7.4 by the broth microdilution method, even though quality control results indicated valid testing at both pHs . This study indicated that the results of tests of the susceptibility of H . influenzae with clarithromycin and azithromycin are highly dependent on the pH of the medium . Test results and their interpretations varied even when the medium pH was within the NCCLS-approved range and, coupled with the current NCCLS breakpoint of 8 microg/ml in the case of clarithromycin, may explain some of the observed discordances between the disk diffusion and broth microdilution methods.

J Biol Chem, 1997 May 23, 272(21), 13614 - 21
Porins of Haemophilus influenzae type b mutated in loop 3 and in loop 4; Srikumar R et al.; Porin (341 amino acids; mass of 37,782 Da) in the outer membrane of Haemophilus influenzae type b (Hib) permits diffusion into the periplasm of small solutes up to a molecular mass of 1400 Da . Molecular modeling of Hib porin identified its structural similarities to OmpF of Escherichia coli and disclosed for Hib porin a shorter length of loop 3 and a longer length of loop 4 . By site-directed mutagenesis of the porin gene ompP2, mutant porins were constructed to contain 6 or 12 amino acid deletions either in loop 3 or in surface-exposed loop 4 . Wild type Hib porin and mutant porins were expressed in a nontypeable H . influenzae strain deleted for the ompP2 gene . The mutant porins were purified and reconstituted into planar bilayers, tested for channel formation and compared with wild type Hib porin . Mutant Haemophilus porin possessing a 6-amino acid deletion in loop 3 displayed a broad distribution of single channel conductance values, while deletion of 12 amino acids from the same loop destabilized the porin channel . By comparison, deletion of 6 or of 12 amino acids from loop 4 of Hib porin resulted in an increased single channel conductance (1.15 and 1.05 nanosiemens, respectively) compared with wild type Hib porin (0 . 85 nanosiemens) . The C3 epitope of the poliovirus VP1 capsid protein was inserted either into loop 3 or into loop 4 of Hib porin . By flow cytometry, the C3 epitope was detected as surface-exposed in strains expressing C3 insertion in loop 4; in strains expressing C3 insertion in loop 3, the epitope was inaccessible . We propose that loop 4 of Hib porin, although surface-accessible, is oriented toward the central axis of the pore and that deletions in this loop increase the single channel conductance by widening the pore entrance.

Gene, 1997 May 20, 191(1), 57 - 60
Sequence variation in the hpd gene of nonencapsulated Haemophilus influenzae isolated from patients with chronic bronchitis; Duim B et al.; The molecular diversity of protein D of nonencapsulated Haemophilus influenzae strains isolated from persistently infected patients with chronic bronchitis was studied by sequencing the hpd gene of four independently obtained isolates . The nucleotide (nt) sequences of the hpd genes of two strains were identical . The other two hpd sequences showed nt substitutions which were mostly synonymous . As a consequence the deduced amino acid (aa) sequences differed from the consensus sequence only by a few aa . No changes in the hpd genes were observed among the four variants of the four strains persisting in chronic bronchitis patients for 9, 11, 8 and 3 months, respectively, although variation in their major outer membrane proteins P2 and P5 occurred . We conclude that the hpd gene is conserved during chronic infections of nonencapsulated H . influenzae.

Biochemistry, 1997 May 6, 36(18), 5509 - 21
Membrane localization, topology, and mutual stabilization of the rnfABC gene products in Rhodobacter capsulatus and implications for a new family of energy-coupling NADH oxidoreductases; Kumagai H et al.; The rnf genes in Rhodobacter capsulatus are unique nitrogen fixation genes that encode potential membrane proteins (RnfA, RnfD, and RnfE) and potential iron-sulfur proteins (RnfB and RnfC) . In this study, we first analyzed the localization and topology of the RnfA, RnfB, and RnfC proteins . By activity and immunoblot analysis of expression of translational fusions to Escherichia coli alkaline phosphatase, RnfA protein was shown to span the chromatophore membrane with its odd-numbered hydrophilic regions exposed to periplasm . By alkaline treatment of membrane fractions and following immunoblot analysis using antibodies against recombinant proteins expressed in E . coli, both RnfB and RnfC proteins were revealed to situate at the periphery of the chromatophore membranes . Second, mutual interaction of the Rnf proteins was analyzed by immunochemical determinations of RnfB and RnfC proteins in rnf mutants and their complemented derivatives . The contents in cellular fractions indicated that RnfB and RnfC stabilize each other and that the presence of RnfA is necessary for stable existence of both proteins . These results support a hypothesis that the Rnf products are subunits of a membrane complex . Finally, we detected homologs of rnf genes in Haemophilus influenzae and Vibrio alginolyticus by data base searches and in E . coli by cloning of a fragment of an rnfA homolog followed by a data base search . Close comparisons revealed that RnfC has potential binding sites for NADH and FMN which are similar to those found in proton-translocating NADH:quinone oxidoreductases and that RnfA, RnfD, and RnfE show similarity to subunits of sodium-translocating NADH:quinone oxidoreductases . We predict that the putative Rnf complex represents a novel family of energy-coupling NADH oxidoreductases.

Tierarztl Prax, 1997 May, 25(3), 226 - 32
{Examination of primary SPF swine after experimental infection with Haemophilus parasuis . Clinical symptoms, changes in hematological parameters and in the parameters of the cerebrospinal fluid}; Wiegand M et al.; The objectives of this work were to cause the Glasser's disease (GD) in primary specific pathogen free piglets after experimental infection, to observe the clinical symptoms and to examine the influence of the infection on the haematological parameters . GD was caused by experimental infection of Haemophilus parasuis in seven to eight weeks old specific pathogen free piglets . In relation to the infection route the morbidity was high (83-100%) and 20% of the infected piglets died . Based on the physical examination fever, respiratory distress, cramps and paralysis were observed which are typical for GD . Arthritis and nerval symptoms are also typical but less common in Glasser's disease . PCV was significantly decreased and WBC significant increased before the piglets were euthanatized.

Radiol Med (Torino), 1997 May, 93(5), 532 - 8
{Diagnostic imaging and therapeutic implications in lung infections in patients with HIV-1 infection}; Carella E et al.; We studied retrospectively 132 episodes of infectious pneumonias in 89 patients examined from 1990 to 1995 . Pneumocystis carinii was found to be the most common cause of pneumonia (33 patients) . The other causes were: Streptococcus pneumoniae (15), Mycobacterium tuberculosis (14), Pseudomonas aeruginosa (8), Staphylococcus aureus (5), Cytomegalovirus (4), Haemophilus influentiae (4), Mycobacterium avium intracellulare (2), Klebsiella pneumoniae (2), E . coli (2), Serratia marcescens (1) . No etiologic agent was found in 40 cases . We stress the need of a more frequent use of invasive diagnostic procedures in the study of focal lung consolidations because this radiologic sign is highly aspecific and may be caused by too many different pathogenic agents, needing different therapies-i.e., Streptococcus pneumoniae (15 cases), Pseudomonas aeruginosa (8), Staphylococcus aureus (5), Klebsiella pneumoniae (2), Escherichia coli (2), Pneumocystis carinii, Serratia marcescens and Haemophilus influentiae (1) . Since there is an increase in mortality among patients treated with empiric antibiotic therapy, we stress the need of the routinary use of bronchoalveolar lavage in HIV+ patients with lung consolidation to perform specific therapy . Moreover, Pneumocystis carinii is by far the most frequent cause of diffuse interstitial infiltrates, and PCP has very suggestive clinical (dyspnea), radiologic (diffuse perihilar interstitial infiltrates; ground glass opacities; pneumatoceles) and laboratory (CD3+CD4 < 200/mcl; LDH > 600 UI/dl; PO2 < 70 mmHg) patterns, always related to the discovery of Pneumocystis carinii in escreatum . Thus, we decided to treat 15 patients with specific therapy for Pneumocystis carinii pneumonia with the above diagnostic algorithm, obtaining in all of them complete clinical and radiologic recovery . To conclude, in critical patients, invasive procedures should be performed only in the cases in which PCP is clinically improbable.

Radiol Med (Torino), 1997 May, 93(5), 520 - 6
{Slow-resolution pneumonia: radiologic and tomodensitometric features}; Polverosi R et al.; To assess the radiologic findings of unresolving pneumonias (radiologic signs not normalized at the end of the fourth week of antibiotic therapy), the radiologic (192) and CT examinations (33) of 50 patients were reviewed, for a total of 64 pulmonary parenchymal lesions . These lesions presented as masses (3; 4.6%), parenchymal thickenings with aerial bronchogram (41; 63.7%), sometimes escavated (4; 10%), alveolar ground glass alterations (4; 6.2%), more frequent on the right side (49; 73%), mostly in the upper lobe (21; 32.8%) . Pleural effusion was demonstrated in 16 patients (32%) and lymphadenopathies were shown in 8 patients (8%) . When possible, the responsible germ was isolated with microbiological expectoration test (14; 35.3%), bronchoscopy associated with bronchoalveolar lavage (7; 37.5%) and needle biopsy in the mass (2; 5%) . The germs were Candida (9), haemophilus influentiae (1), Klebsiella (1), pneumococcus (1), Pseudomonas (3), Staphylococcus aureus (4), streptococcus alpha hemoliticus (6) . There are no typical radiologic and CT findings for the described lesions in different-patients and different germs can produce the same changes . Therefore, pulmonary inflammations must be followed to their complete resolution, to rule out the chance of a neoplastic process (obstructive pneumonia) . The first radiologic control must be made at least 15 days after the beginning of therapy for the anatomo-pathologic processes of recovery to show radiographically . CT can be fundamental in the differential diagnosis with neoplasm, in unresolving pneumonia, to study the tracheobronchial tree, to study lymphadenopathies and to guide needle biopsy.

J Chemother, 1997 May, 9 Suppl 3, 38 - 44
Relevance of pharmacokinetics and pharmacodynamics in the selection of antibiotics for respiratory tract infections; Drusano GL et al.; The pharmacodynamic principles that link the concentrations of antibiotics within body systems and their effects have been elucidated only recently . Animal work, now confirmed by clinical studies, has shown that for beta-lactam antibiotics, the time that the serum concentration exceeds the minimum inhibitory concentration (MIC) value of the pathogen is a key parameter in predicting a successful clinical and bacteriological outcome . The situation with the macrolides is less clear; time above MIC is the dynamic variable likely to be most closely linked to efficacy for erythromycin and clarithromycin but for azithromycin it appears to be the area under the plasma concentration-time curve: MIC ratio . Different antibiotics are appropriate for the key pathogens in community-acquired respiratory tract infections . For Streptococcus pneumoniae, amoxycillin/clavulanate is effective with varying dosage regimens providing around 40% time above the MIC90, in contrast to the oral cephalosporins and the macrolides for which serum concentrations do not exceed the MIC90 . For Haemophilus influenzae, amoxycillin/clavulanate and cefixime are suitable antibiotics whereas macrolides have limited activity . With the exception of amoxycillin, all the beta-lactam and macrolide antibiotics reviewed here perform better against Moraxella catarrhalis than against the other two principal community-acquired respiratory tract pathogens and there is a wide choice of appropriate agents . Knowledge of the pharmacodynamically-linked variables for different antibiotics allows optimization of dosage regimens and direct comparisons across agents for the same variables.

J Chemother, 1997 May, 9 Suppl 3, 18 - 28
Trends in the activity of macrolide and beta-lactam antibiotics and resistance development . Alexander Project Group; Schito GC et al.; The Alexander Project is an ongoing international multicenter study monitoring trends in the antimicrobial susceptibilities of community-acquired lower respiratory tract (LRT) pathogens . In 1995, 4011 isolates were collected . The incidence of beta-lactamase-positive Haemophilus influenzae was 28.4% in the United States and 15.4% in Europe, and the incidence of beta-lactamase-positive Moraxella catarrhalis has risen to > 90% in Europe and the United States . The incidence of penicillin-resistant Streptococcus pneumoniae is higher in Europe (24.9%) than the Unites States (12.3%) . For the majority of centers, there is a marked association between penicillin and macrolide resistance in S . pneumoniae with erythromycin, azithromycin and clarithromycin exhibiting MIC90s of > or = 32 mg/l against penicillin-resistant strains . For Toulouse and Genoa, at least, the high levels of macrolide resistance may be attributable to high macrolide usage . Ceftriaxone and amoxycillin/clavulanate are the most potent agents for empirical therapy, with MIC90s of < or = 2 mg/l against all three principal pathogens . The majority of oral agents studied are active against > 90% H . influenzae and M . catarrhalis and > 80% S . pneumoniae on breakpoint criteria . However, on the basis of the time above MIC criteria for the beta-lactam and macrolide agents tested, only amoxycillin/clavulanate and the parenteral agent ceftriaxone can be recommended for empirical therapy of LRT infections caused by these pathogens.

J Chemother, 1997 May, 9 Suppl 3, 10 - 7
Respiratory tract infection: epidemiology and surveillance; Jacobs MR; Streptococcus pneumoniae and Haemophilus influenzae are the key pathogens implicated in bacterial infections of the upper and lower respiratory tract . Choice of empiric oral antimicrobial chemotherapy is guided by the clinical presentation, severity of the infection and epidemiological knowledge . beta-Lactams and the macrolides are the two major groups of antibiotics used to treat respiratory tract infections . The prevalence of penicillin-resistant strains of S . pneumoniae is increasing world-wide (up to 30% in the USA), as is the prevalence of beta-lactamase-producing strains of H . influenzae . Macrolide resistance in S . pneumoniae is increasing and is absolute, and some of the macrolides have only limited activity against H . influenzae . Knowledge of local and global antibiotic resistance patterns should be used as the key to directing empiric choice of antibiotic treatment.

J Chemother, 1997 May, 9 Suppl 3, 5 - 9
Empiric therapy in lower respiratory tract infection--an ongoing challenge; Geddes AM; Lower respiratory tract infections are a common cause of morbidity and mortality . The pattern of pneumonia is altering, owing to changes in a number of influencing factors . These include patient characteristics, such as an aging population, increased immunosuppression and chronic disease, and changes in medical practice . There is also an increasing level of resistance to antimicrobial agents by organisms such as Streptococcus pneumoniae and Haemophilus influenzae, the pathogens most commonly associated with community-acquired pneumonia . In the management of pneumonia, it is important to be able to differentiate between atypical and typical pneumonia in the clinical setting and to grade the severity of the infection . Currently, there are no internationally agreed treatment recommendations for pneumonia . The role of antimicrobial agents in acute exacerbations of chronic bronchitis is still a controversial issue.

Vet Microbiol, 1997 May, 56(1-2), 125 - 34
Analysis of Haemophilus parasuis by multilocus enzyme electrophoresis; Blackall PJ et al.; The diversity among 40 Australian isolates and eight reference strains of Haemophilus parasuis was examined using multilocus enzyme electrophoresis . Thirty-four electrophoretic types (ETs) were recognized with a mean diversity per locus of 0.405 . One Australian isolate was located in an ET separated by a considerable distance (> 0.8) from the rest of the isolates, suggesting that it may represent a different species or subspecies . The remaining 33 ETs formed two distinct divisions (A and B), separated from each other by a distance of 0.506 . All 12 Australian isolates of serovar 5 plus the two reference strains for this serovar were included in Division A . The only other isolates present in this Division were Australian isolates of serovars 4 and 13 and two nontypeable isolates . Division B contained a diverse range of serovars-Australian isolates of serovars 1, 2, 7/10, 9 and 13 as well as the reference strains for serovars 1, 2, 3, 4, 8 and 9 . These results supported other studies which demonstrated considerable diversity, and indicate that the population of H . parasuis may contain more than a single species or subspecies . There was considerable diversity even amongst isolates of the same serovar, indicating that serotyping is not a particularly suitable technique for strain typing in epidemiological studies.

Eur J Clin Microbiol Infect Dis, 1997 May, 16(5), 391 - 4
Comparison of agar dilution, broth dilution, disk diffusion, and the E-test for susceptibility testing of penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae; Thorvilson J et al.; An evaluation to determine the optimal methods for the in vitro susceptibility testing of 41 clinical isolates and the ATCC 49619 strain of Streptococcus pneumoniae to penicillin was undertaken . No very major or major interpretive errors were observed with the following test methods and media: agar dilution using either Mueller-Hinton medium with lysed horse blood or Haemophilus test medium; broth dilution using cation-adjusted Mueller-Hinton medium with lysed horse blood, Haemophilus test medium, or Todd-Hewitt medium; and the epsilo-meter test (E-test) using agar containing Mueller-Hinton medium and 5% sheep blood . The disk diffusion method using agar containing Mueller-Hinton medium and 5% sheep blood agar was an effective screening method, requiring confirmation by a dilution susceptibility test method.

Eur J Clin Microbiol Infect Dis, 1997 May, 16(5), 377 - 80
Invasive Haemophilus influenzae in the Republic of Ireland; Moloney AC et al.; Prior to the general availability of Haemophilus influenzae type b vaccine in the Republic of Ireland, a two-year study of the epidemiology of invasive Haemophilus influenzae disease was carried out . Of 137 invasive strains of Haemophilus influenzae examined in a central laboratory, 94.2% were serotype b and 90.5% were biotype I . Seventeen percent of serotype b strains produced beta-lactamase, and 2.3% were resistant to both ampicillin and chloramphenicol . The majority of serotype b strains were electrophoretic types of the electrophoretic 12 clone family, principally 12.5 . Meningitis was the most common infection caused by serotype b . The study data extend the current knowledge of strains of Haemophilus influenzae causing invasive disease in the Republic of Ireland.

Jpn J Antibiot, 1997 May, 50(5), 421 - 59
{Susceptibilities of bacteria isolated from patients with respiratory infectious diseases to antibiotics (1995)}; Ikemoto H et al.; The bacteria isolated from the patients with lower respiratory tract infections were collected by institutions located throughout Japan, since 1981 . Ikemoto et al . have been investigating susceptibilities of these isolates to various antibacterial agents and antibiotics, and characteristics of the patients and isolates from them each year . Results obtained from these investigations are discussed . In 23 institutions around the entire Japan, 567 strains of presumably etiological bacteria were isolated mainly from the sputa of 459 patients with lower respiratory tract infections during the period from October 1995 to September 1996 . MICs of various antibacterial agents and antibiotics were determined against 74 strains of Staphylococcus aureus, 82 strains of Streptococcus pneumoniae, 104 strains of Haemophilus influenzae, 85 strains of Pseudomonas aeruginosa (non-mucoid strains), 18 strains of Pseudomonas aeruginosa (mucoid strains), 52 strains of Moraxella subgenus Branhamella catarrhalis, 25 strains of Klebsiella pneumoniae etc., and the drug susceptibilities of these strains were assessed except for those strains that died during transportation . 1) S . aureus . S . aureus strains for which MICs of oxacillin (MPIPC) were higher than 4 micrograms/ml (methicillin-resistant S . aureus) accounted for 52.7% . Arbekacin (ABK) showed the most highest activity against S . aureus with MIC80 of 0.5 micrograms/ml . Vancomycin (VCM) showed the next highest activity with MIC80 of 1 microgram/ml . These drugs showed the high activities against MRSA with MIC80S of 1 microgram/ml . 2) S . pneumoniae . Most of drugs tested showed potent activities against S . pneumoniae . Imipenem (IPM) and panipenem (PAPM), carbapenems, showed the most potent activity with MIC80S of 0.063 microgram/ml . Cefotaxime (CTX), cefmenoxime (CMX) and cefpirome (CPR) of cephems showed the next most potent activities with MIC80S of 0.25 microgram/ml . Erythromycin (EM) and clindamycin (CLDM) showed low activities with MIC80S 128 micrograms/ml or high . Among these strains, however, 48.8% and 65.9% of respective strains were quite toward sensitive these agents with MICs of 0.063 microgram/ml . 3) H . influenzae . The activities of all drugs were potent against H . influenzae test with all MICs at 4 micrograms/ml or below . Cefotiam (CTM), CMX, cefditoren (CDTR) and ofloxacin (OFLX) showed the most potent activity with MIC90S to 0.063 microgram/ml . 4) P . aeruginosa . (mucoid strains) IPM and tobramycin (TOB) showed the most potent activity against P . aeruginosa (mucoid strains) with MIC80S of 1 microgram/ml . Ceftazidime (CAZ), cefsulodin (CFS) and carumonam (CRMN) showed next potent activity, with MIC80S of 2 micrograms/ml . The MIC80S of the other drugs ranged from 4 micrograms/ml to 32 micrograms/ml . 5) P . aeruginosa (non-mucoid strains) . TOB and ciprofloxacin (CPFX) showed the most potent activities against P . aeruginosa (non-mucoid strains) with MIC80S of 1 microgram/ml . The MIC80 of ampicillin (ABPC) was 128 micrograms/ml in 1994, it was 16 micrograms/ml in 1995 . 6) K . pneumoniae . All drugs except ABPC were active against K . pneumoniae . CPR and CRMN showed the most potent activities against K . pneumoniae with MIC80S of 0.063 microgram/ml . The MIC80S of the other drugs ranged from 0.125 microgram/ml to 2 micrograms/ml . 7) M . (B.) catarrhalis . Against M . (B.) catarrhalis, all the drugs showed good activities with MIC80S at 4 micrograms/ml or below . And MICs of all strains were 8 micrograms/ml or below . IPM, OFLX and minocycline (MINO) showed the most potent activity with MIC80S of 0.063 microgram/ml . Also, we investigated year to year changes in the characteristics of patients, their respiratory infectious diseases, and the etiology . Patients' backgrounds were examine for 567 isolates from 459 cases . The examination of age distribution found that the proportion of patients with ages over 60 years was 66.3% of all the patients showing a slight increase over that in 1994 . Proportion of differe

Kansenshogaku Zasshi, 1997 May, 71(5), 430 - 6
{The evaluation of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) level in peripheral blood of patients with chronic lower respiratory tract infection}; Tsujimoto M et al.; In the present study, we assessed the serum level of IL-6 and TNF-alpha by ELISA in patients with chronic lower respiratory tract infection . The serum levels of IL-6 and TNF-alpha of patients in acute exacerbation phase are higher than that of in stable phase . We also classified patients in acute exacerbation phase into two groups according to the microorganism of persistent infection . The serum level of IL-6 and TNF-alpha in the patients with persistent infection with Pseudomonas aeruginosa were higher than that with Haemophilus influenzae . Moreover, the serum level of IL-6 and TNF-alpha were found to be related with malnutrition which assessed by clinical indices such as the serum level of albumin and cholinesterase . The present result suggests that IL-6 and TNF-alpha may have relationship with not only inflammation in airway but also indices of nutrition in patients with chronic lower respiratory tract infection.

Arzneimittelforschung, 1997 May, 47(5), 674 - 7
Cefodizime once daily in the treatment of lower respiratory tract infections; Piovano CF et al.; Cefodizime (CAS 69739-16-8, HR 221) is a new third-generation cephalosporin with pharmacokinetic properties that make it suitable for once-daily administration in the treatment of lower respiratory tract infections (LRTI) . Ninety-nine adult hospitalized patients (66 males, 33 females, median age 57.5 years) received a once-daily injection of 2 g cefodizime for LRTI . Median treatment duration was 8 days . Forty-two patients received cefodizime intravenously and 57 intramuscularly . Indications for treatment were as follows; primary lobar pneumonia (n = 36), bronchopneumonia (n = 14), secondary pneumonia (n = 3), aspiration pneumonia (n = 5), acute exacerbation of chronic bronchitis (n = 21), and of bronchiectasis (n = 9) and acute purulent bronchitis (n = 11) . General condition was good in 29 patients and poor in 58; 12 patients were critically ill . The following pathogens were isolated at baseline (source: bronchial secretions, sputum or blood): S . pneumoniae (n = 47), Haemophilus spp . (n = 17), M . catarrhalis (n = 6), Streptococcus spp . (n = 9), Staphylococcus spp . (n = 5), Klebsiella spp . (n = 4), Pseudomonas spp . (n = 1), A . calcoaceticus (n = 1) and anaerobic organisms (n = 7) . Fifty-nine patients were evaluable for bacteriological response and 82 for clinical response . Bacteriological outcome was satisfactory in 29/30 patients having LRTI with parenchymal involvement (97%) and in 29/29 patients without parenchymal involvement (100%) . Clinical cure was achieved in 41/43 evaluable patients with parenchymal involvement (95%) and in 37/39 patients without parenchymal involvement (95%) in the per-protocol analysis and in 54/58 patients (93%) and 37/41 patients (93%), respectively, in the clinical intention-to-treat analysis . Three of the patients with an unsatisfactory clinical response died of infection during the study . Cefodizime was well tolerated . Adverse reactions--all of mild intensity--were tachycardia, lumbalgia and dizziness, each occurring in one patient . Cefodizime 2 g once daily either i.m . or i.v . was effective in the treatment of lower respiratory tract infections in hospitalized patients.

Acta Otolaryngol, 1997 May, 117(3), 396 - 405
The tympanic membrane and middle ear mucosa during non-typeable Haemophilus influenzae and Haemophilus influenzae type b acute otitis media: a study in the rat; Magnuson K et al.; Non-typeable Haemophilus influenzae (NTHi) and encapsulated Haemophilus influenzae type b (Hib) were inoculated into the middle ears of Sprague-Dawley rats . Tympanic membrane (TM) status was assessed otomicroscopically and specimens from various middle ear areas were prepared for light microscopy at various times during the acute phase and up to 6 months after inoculation . Irrespective of bacteria strain, acute otitis media (AOM) was present in all ears 4 days after inoculation . The Hib-infected ears showed initially a severe course of AOM, but all were otomicroscopically resolved by day 12, at which time a few NTHi-inoculated ears still exhibited middle ear effusion . The TMs infected with Hib had normalized without scar formation, whereas NTHi induced a persistent thickening of the TMs in half of all cases . The middle ear mucosa of NTHi-infected ears initially showed vigorous activity among the goblet cells, but the mucosa normalized after the acute phase . Hib, by contrast, induced prominent changes in the middle ear mucosa . Initially, no goblet cell granules or ciliated cells could be observed in the mucosa . Later on, the epithelium contained large, active goblet cells . Glands appeared beneath the mucosa which persisted as streaks of epithelial cells throughout the study period . The findings show that NTHi and Hib both induce AOM but with differing clinical courses, and affect different targets in the middle ear.

Acta Otolaryngol, 1997 May, 117(3), 352 - 7
Electrophysiological effects of multiple instillation of Haemophilus influenzae type b endotoxin on the inner ear; Stenqvist M et al.; An analysis of auditory brainstem response (ABR) thresholds and ABR-based frequency tuning curves was performed in 15 Sprague-Dawley rats exposed to Haemophilus influenzae type b endotoxin; 5 microg/50 microl toxin was instilled every second day, altogether five times, into the middle ear cavity through a small perforation in the tympanic membrane . ABR was measured 48 h after the second application and 24 h, 48 h, 5 days and 10 days after the fifth instillation . Five applications of toxin had no statistical effect on ABR thresholds and no changes in TC configuration were observed . It is concluded that Haemophilus influenzae type b endotoxin, instilled repeatedly through the tympanic membrane into the middle ear, does not affect cochlear electrophysiology.

Mol Microbiol, 1997 May, 24(4), 767 - 78
EspP, a novel extracellular serine protease of enterohaemorrhagic Escherichia coli O157:H7 cleaves human coagulation factor V; Brunder W et al.; In this study, we identified and characterized a novel secreted protein, the extracellular serine protease EspP, which is encoded by the large plasmid of enterohaemorrhagic Escherichia coli (EHEC) O157:H7 . The corresponding espP gene consists of a 3900 bp open reading frame that is able to encode a 1300-amino-acid protein . EspP is synthesized as a large precursor which is then processed at the N- and C-termini during secretion . It can be grouped into the autotransporter protein family . The deduced amino acid sequence of EspP showed homology to several secreted or surface-exposed proteins of pathogenic bacteria, in particular EspC of enteropathogenic E . coli and IgA1 proteases from Neisseria spp . and Haemophilus influenzae . Hybridization experiments and immunoblot analysis of clinical EHEC isolates showed that EspP is widespread among EHEC of the serogroup O157 and that it also exists in serogroup 026 . A specific immune response against EspP was detected in sera from patients suffering from EHEC infections . Functional analysis showed that EspP is a protease capable of cleaving pepsin A and human coagulation factor V . Degradation of factor V could contribute to the mucosal haemorrhage observed in patients with haemorrhagic colitis.

Exp Neurol, 1997 May, 145(1), 253 - 7
Blood-brain barrier permeability during the development of experimental bacterial meningitis in the rat; Kim KS et al.; In an attempt to examine whether routes of bacterial entry into the central nervous system have any bearing on subsequent changes in blood-brain barrier permeability, we examined cerebrospinal fluid (CSF) penetration of circulating 125I-albumin in two different models of experimental meningitis due to K1 Escherichia coli, type III group B streptococcus, or Haemophilus influenzae type b in infant rats: hematogenous meningitis subsequent to subcutaneous inoculation of bacteria vs meningitis induced by direct inoculation of bacteria into the CSF via the cisterna magna . In the model of hematogenous meningitis, the mean CSF penetration was significantly greater in animals with H . influenzae type b meningitis than in those with meningitis due to K1 E . coli or type III group B streptococcus . In contrast, the mean CSF penetration was significantly enhanced in all animals with meningitis induced by intracisternal inoculation regardless of infecting pathogens . Tumor necrosis factor activity in CSF appeared to correlate with the functional penetration of circulating albumin across the blood-brain barrier in both models of experimental meningitis . These findings suggest that the alterations of blood-brain barrier permeability during development of experimental meningitis may vary for different models of inducing meningitis and that the mechanisms responsible for these different permeability changes may be multifactorial.

Eur J Biochem, 1997 May 1, 245(3), 790 - 6
Purification and characterization of flavoproteins and cytochromes from the yellow bioluminescence marine bacterium Vibrio fischeri strain Y1; Petushkov VN et al.; Several flavoproteins and cytochromes that occur as major components in extracts of the yellow bioluminescence Y1 strain of the marine bacterium Vibrio fischeri have been purified and characterized with respect to their mass (SDS/PAGE and matrix-assisted laser-desorption/ionization MS), chromatographic properties, N-terminal sequence, and spectroscopy (absorption, fluorescence emission and anisotropy decay) . The investigated proteins were as follows: yellow fluorescence protein (YFP) with bound riboflavin, FMN or 6,7-dimethyl-8-ribityllumazine; a blue fluorescence protein (BFP) with bound 6,7-dimethyl-8-ribityllumazine, riboflavin, or 6-methyl-7-oxo-8-ribityllumazine; thioredoxin reductase with FAD as ligand; and two c-type diheme cytochromes, c551 and c554 . We present evidence that the riboflavin-bound YFP has an N-terminal sequence corresponding to that published for the dimeric YFP . We show that an equilibrium replacement of the riboflavin can be made with excess lumazine derivative and that lumazine-bound YFP has different bioluminescence properties to those of the lumazine protein from Photobacterium leiognathi . BFP is a different protein again, and in the bacterial lysate it occurs in multiple forms, ligated to either riboflavin, lumazine, or the 7-oxolumazine derivative . The N-terminal sequence for BFP shows similarities to those of the YFP proteins and to lumazine protein and riboflavin synthase from Photobacterium . BFP in any form has no bioluminescence or riboflavin-synthase activity . A 70-kDa fluorescent flavoprotein with FAD as ligand has an N-terminal sequence highly similar to those of thioredoxin reductases from Haemophilus influenzae and Escherichia coli . Cytochrome contaminations in previous preparations of YFP have been removed and are identified as the two c-type cytochromes c551 and c554 . Both inhibit the NADH-induced bioluminescence in the reductase/luciferase system with the luciferases from P . leiognathi and V . fischeri . The N-terminal amino acid sequence of the cytochrome (c551) corresponds to a diheme cytochrome c4 . The spectral properties of c554 are similar to those of other c5 cytochromes, and both c554 and c551 have absorption spectra similar to those of the respective cytochromes from the gram-negative bacteria Pseudomonas and Azotobacter.

Microbiology, 1997 May, 143 ( Pt 5), 1709 - 16
Molecular and immunological characterization of OprL, the 18 kDa outer-membrane peptidoglycan-associated lipoprotein (PAL) of Pseudomonas aeruginosa; Lim A Jr et al.; Immunological screening of a Pseudomonas aeruginosa cosmid library led to the identification of clones producing an 18 kDa outer-membrane protein . This protein reacted in Western blots with a polyclonal antiserum against outer-membrane proteins of P . aeruginosa and with a monoclonal antibody (MA1-6) specific for OprL, the peptidoglycan-associated outer-membrane lipoprotein (PAL) . Sequencing of pOML7, a subclone expressing oprL, revealed an ORF of 504 bp encoding a polypeptide with a typical lipoprotein signal recognition sequence . Another ORF was found upstream of oprL, with homology to the TolB protein of Escherichia coli and Haemophilus influenzae . Downstream of oprL, a second ORF, of 321 bp, was found (orf2), encoding a protein with a signal peptide and with no homology with proteins of known biological function . After the stop codon of orf2, a rho-independent terminator sequence was detected which is part of the P . aeruginosa PAO1 insertion element IS222 . OprL showed homologies with all known PALs from Gram-negative bacteria, especially in the C-terminal part . mAb MA1-6 reacted with P . aeruginosa cells in immunofluorescence, and with E . coli cells expressing oprL, which had an abnormal, elongated morphology, an indication that production of the protein perturbed the division process.

FEMS Microbiol Lett, 1997 May 1, 150(1), 19 - 26
Elemental iron does repress transferrin, haemopexin and haemoglobin receptor expression in Haemophilus influenzae; Hasan AA et al.; The iron repressible nature of Haemophilus influenzae transferrin binding proteins suggests a regulatory role for elemental iron in their expression . The existence of a Haemophilus ferric uptake repressor (Fur) binding motif identified in the promoter region of both tbpA and tbpB further supports this hypothesis . However, a recent study using brain heart infusion growth medium suggested that transferrin binding protein synthesis in H . influenzae was haem-rather than iron-regulated . The present study re-investigates this observation and using a chemically defined medium, we demonstrate that elemental iron haem or protoporphyrin IX can each regulate Haemophilus influenzae transferrin, haemopexin and haemoglobin receptor expression.

Eur Respir J, 1997 May, 10(5), 1137 - 44
Bacterial colonization of distal airways in healthy subjects and chronic lung disease: a bronchoscopic study; Cabello H et al.; In contrast to the healthy population, distal airway bacterial colonization may occur in patients with chronic lung diseases, who often have altered pulmonary defences . However, the information dealing with this issue is insufficient and is based mainly on nonspecific samples, such as sputum cultures . Using quantitative cultures of bronchoscopic protected specimen brush (PSB) and bronchoalveolar lavage (BAL) samples, we studied the bacterial colonization of distal airways in 16 healthy subjects, 33 patients with bronchogenic carcinoma, 18 with chronic obstructive pulmonary disease (COPD), 17 with bronchiectasis, and 32 with a long-term tracheostomy due to laryngeal carcinoma . All patients were without exacerbation, and free from antibiotic treatment at least 1 month before the study protocol . Thresholds for quantitative cultures to define colonization were > or = 10(2) colony-forming units (cfu) x mL(-1) for PSB and > or = 10(3) cfu x mL(-1) for BAL . Only one healthy subject was colonized by a potential pathogenic microorganism (PPM) (Staphylococcus aureus 4x10(2) cfu x mL(-1) in a PSB culture) . Colonization was observed in 14 (42%) bronchogenic carcinoma patients (19 non-PPMs, and 10 PPMs); in 15 (83%) COPD patients (22 non-PPMs and 7 PPMs); in 15 (88%) bronchiectasis patients (20 non-PPMs and 13 PPMs); and in 15 (47%) long-term tracheostomy patients (5 non-PPMs and 13 PPMs) . The two most frequent non-PPMs isolated in all groups studied were Streptococcus viridans and Neisseria spp . Haemophilus spp., Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis were the most frequent PPMs isolated in bronchogenic carcinoma, COPD, bronchiectasis and long-term tracheostomized patients, respectively . Pseudomonas aeruginosa colonization was infrequent in all the groups . Our results show that distal airway bacterial colonization is a frequent feature in stable patients with chronic lung diseases and also in patients with long-term tracheostomy . However, the pattern of colonization differs among groups studied . The knowledge of different colonization patterns may be important for future antibiotic prophylactic strategies and for the empirical antibiotic regimens when exacerbations occur in these patients.

Pediatr Infect Dis J, 1997 May, 16(5), 471 - 8
Cefprozil treatment of persistent and recurrent acute otitis media; Pichichero ME et al.; OBJECTIVE: We identified the pathogens causing persistent and recurrent acute otitis media (AOM) and the clinical efficacy of cefprozil as treatment . STUDY DESIGN: This was a noncomparative, open label multicenter trial . Children ages 6 months to 12 years with signs and symptoms of AOM and evidence of middle ear effusion, as confirmed by pneumatic otoscopy or tympanometry, underwent tympanocentesis and subsequent treatment with cefprozil (15 mg/kg given twice daily) for 10 days . Patients with recurrent otitis media or failure of previous antibiotic therapy or prophylaxis were particularly sought for the study . RESULTS: Two hundred sixty-two (99%) of 265 enrolled children were considered evaluable . The median age of the study group was 1 year . Ninety-eight (37%) of the children had a history (within 30 days) of prior antibiotic use . Ninety-seven (37%) met our definition of recurrent AOM, 48 (18%) met our definition of persistent AOM and 132 (50%) children had 3 or more previous episodes of acute otitis media within 12 months before study . Eighty-two (31%) of the enrollment tympanocentesis had no growth, 150 (57%) had a single bacterial pathogen and 29 (11%) had multiple bacterial pathogens . Of the 93 Streptococcus pneumoniae pretreatment isolates, 50 (54%) were penicillin-susceptible, 12 (13%) were penicillin-intermediate resistant and 31 (33%) were penicillin-resistant . Of the 75 Haemophilus influenzae pretreatment isolates, 42 (56%) produced beta-lactamase as did 4 (27%) of the 15 Moraxella catarrhalis strains . A satisfactory clinical response by pathogen was found in 75% (70 of 93) with S . pneumoniae, 75% (56 of 75) with H . influenzae and 93% (13 of 14) with M . catarrhalis; the response with single pathogen infections was higher than those with multiple pathogens (118 of 150 (78%) and 17 of 29 (59%), respectively; P = 0.03) . The response for patients with isolates of S . pneumoniae that were penicillin-susceptible, -intermediate or -resistant were 39 of 50 (78%), 11 of 12 (92%) and 21 of 31 (68%), respectively . Older children had a satisfactory clinical outcome more frequently than younger children (P < 0.001), and the response to therapy varied for persistent, recurrent and recently untreated AOM (P < 0.01) . CONCLUSION: Persistent and recurrent AOM involves the same pathogens as recently untreated AOM but bacteria with reduced antibiotic susceptibility may be more frequently present . This noncomparative study suggests that cefprozil 30 mg/kg/day given in two divided doses for 10 days may be effective in the treatment of children with persistent and recurrent AOM.

Sex Transm Dis, 1997 May, 24(5), 267 - 71
Antibody to Haemophilus ducreyi among trucking company workers in Kenya; Rakwar J et al.; BACKGROUND AND OBJECTIVES: To determine the prevalence, correlates, and incidence of Haemophilus ducreyi antibodies, a cohort of East African trucking company employees was evaluated . STUDY DESIGN: Human immunodeficiency virus (HIV)-1-seronegative men working in six trucking companies in Mombasa, Kenya, were evaluated with a questionnaire and serologic testing for antibodies to H . ducreyi and other sexually transmitted pathogens . Men who were initially H . ducreyi seronegative were retested at 1 year of follow-up . RESULTS: The H . ducreyi seroprevalence among 501 men at enrollment was 26.5% . Seropositivity was significantly associated with older age, married status, years of active sex life, number of sex partners in the past year, history of unprotected sex with a prostitute in the past year, and history of alcohol intake (all P values < 0.01) . Occupational travel for more than 14 days per month was also significantly associated with H . ducreyi seropositivity (odds ratio {OR} 2.1, 95% confidence interval {CI} 1.3-3.2) . Using multivariate analysis, H . ducreyi seropositivity was independently associated with age, married status, history of sex with a prostitute, and history of alcohol intake . Presence of H . ducreyi antibodies was significantly associated with seropositivity to the other major genital ulcerative pathogens, Treponema pallidum (OR 4.3, 95% CI 2.2-8.3), herpes simplex virus type 2 (OR 4.9, 95% CI 2.0-11.5), and Chlamydia trachomatis (OR 3.2, 95% CI 1.5-6.9) . These associations remained significant after adjusting for demographic and exposure variables . The incidence of seroconversion to H . ducreyi antibodies was 3.6 per 100 person years . CONCLUSIONS: Serologic evidence of H . ducreyi infection was common among male trucking company employees . H . ducreyi seropositivity is an objective marker of high-risk behavior and is associated with serologic evidence of other ulcerative sexually transmitted diseasesPIP: A prospective cohort study of 501 human immunodeficiency virus (HIV)-negative male trucking company employees from Kenya revealed high rates of infection with Haemophilus ducreyi, the causative agent of chancroid . At enrollment in March 1993, the seroprevalence of H ducreyi antibodies was 26.5% . Also detected were high rates of herpes simplex virus-2 (49%), Chlamydia trachomatis (41%), and syphilis (8%) . Of the 368 men who were seronegative at enrollment, 241 were re-evaluated after 12 months of follow-up . There were 9 seroconversions (3.6/100 person years) . Sexual contact with a prostitute in the preceding year was reported by 33% of truckers and only a third of these encounters involved condom use . Ever-use of condoms was reported by only 51% . H ducreyi seropositivity was significantly and positively associated with older age, occupational travel for more than 2 weeks per month, history of sex with a prostitute, high number of sex partners in the past year, unprotected sex with a prostitute in the past year, alcohol drinking, and infection with other sexually transmitted diseases . The significant association of H ducreyi and seropositivity to syphilis, herpes simplex virus-2, and C trachomatis (odds ratios: 4.3, 4.9, and 3.2, respectively) raises the possibility that a genital ulcer increases the likelihood of infection with a second ulcerative pathogen . Overall, these findings suggest that the seroprevalence of H ducreyi may be used as an indicator of the extent of high-risk sexual risk behavior in a population, as well as an objective end point for measuring the efficacy of behavioral interventions in communities where the HIV seroincidence is too low to serve this purpose .

J Pediatr, 1997 May, 130(5), 746 - 51
Apnea after immunization of preterm infants; Sanchez PJ et al.; OBJECTIVE: To determine the frequency of adverse reactions, particularly the occurrence of apnea, among preterm infants after immunization with diphtheria and tetanus toxoids and whole cell pertussis vaccine adsorbed (DTP) and Haemophilus influenzae type b conjugate (HibC) vaccine in the neonatal intensive care unit . STUDY DESIGN: After the occurrence of apnea in two preterm infants following immunization with DTP and HibC, a prospective surveillance of 97 preterm infants younger than 37 weeks of gestation who were immunized with DTP (94 also received HibC at the same time) in the neonatal intensive care unit was performed to assess the frequency of adverse reactions and in particular, the occurrence of apnea . For each infant, data were recorded for a 3-day period before and after receipt of the immunization . RESULTS: The majority of preterm infants tolerated immunizations with DTP and HibC without ill effects . However, 12 (12%) infants experienced a recurrence of apnea, and 11 (11%) had at least a 50% increase in the number of apneic and bradycardic episodes in the 72 hours after immunization . This occurred primarily among smaller preterm infants who were immunized at a lower weight (p = 0.01), had experienced more severe apnea of prematurity (p = 0.01), and had chronic lung disease (p = 0.03) . CONCLUSION: The temporal association observed between immunization of preterm infants and a transient increase or recurrence of apnea after vaccination merits further study . Cardiorespiratory monitoring of these infants after immunization may be advisable.

J Bacteriol, 1997 May, 179(9), 3058 - 60
The ispB gene encoding octaprenyl diphosphate synthase is essential for growth of Escherichia coli; Okada K et al.; The Escherichia coli ispB gene encoding octaprenyl diphosphate synthase is responsible for the synthesis of the side chain of isoprenoid quinones . We tried to construct an E . coli ispB-disrupted mutant but could not isolate the chromosomal ispB disrupted mutant unless the ispB gene or its homolog was supplied on a plasmid . The chromosomal ispB disruptants that harbored plasmids carrying the ispB homologs from Haemophilus influenzae and Synechocystis sp . strain PCC6803 produced mainly ubiquinone 7 and ubiquinone 9, respectively . Our results indicate that the function of the ispB gene is essential for normal growth and that this function can be substituted for by homologs of the ispB gene from other organisms that produce distinct forms of ubiquinone.

J Bacteriol, 1997 May, 179(9), 2892 - 9
Identification and characterization of ssb and uup mutants with increased frequency of precise excision of transposon Tn10 derivatives: nucleotide sequence of uup in Escherichia coli; Reddy M et al.; A Lac+ papillation assay was used to identify mutants (tex) of Escherichia coli that exhibit an increased frequency of precise excision of a lacZ::Tn10dKan insertion . Three tex strains had suffered mutations in the gene (ssb) encoding the essential single-stranded DNA-binding protein SSB, which resulted in the following alterations in the 177-residue protein: G4D; L10F, P24S; and V102M . The phenotypes of these ssb mutants indicated that they were largely unaffected in other functions mediated by SSB, such as DNA replication, recombination, and repair . Strains with multicopy ssb+ exhibited a decreased frequency of Tn10dKan precise excision . Three other tex mutants had insertion mutations in the locus designated uup at 21.75 min on the linkage map . The nucleotide sequence of uup was determined, and the gene was inferred to encode a 625-amino-acid hydrophilic protein that belongs to the superfamily of ABC-domain proteins (with two pairs of the Walker A and B motifs), which are postulated to be involved in coupling ATP hydrolysis with other biological processes . The uup gene product shares extensive homology with the deduced sequences of two proteins of Haemophilus influenzae . The uup gene is also situated immediately upstream of (and is transcribed in the same direction as) the paraquat-inducible SoxRS-regulated pqi-5 gene, two reported promoters for which are situated within the uup coding sequence.

Infect Immun, 1997 May, 65(5), 1729 - 33
Brief heat shock treatment induces a long-lasting alteration in the glycolipid receptor binding specificity and growth rate of Haemophilus influenzae; Hartmann E et al.; After brief heat shock treatment, clinical strains of nontypeable Haemophilus influenzae show a long-lasting change in the binding specificity for glycolipids and a markedly increased growth rate in vitro . Non-heat-shocked H . influenzae specifically binds to phosphatidylethanolamine (PE), gangliotetraosylceramide (Gg4), and gangliotriosylceramide (Gg3) and binds minimally to sulfatoxygalactosylceramide (SGC; also called sulfatide) . After a 5-min heat shock at 42 degrees C, strains of H . influenzae showed a marked increase in binding to SGC and acquired the ability to bind to sulfatoxygalactosylglycerol (SGG) in thin-layer chromatography overlays . Additionally, heat-shocked H . influenzae cells showed an increased growth rate (twofold) . Increased sulfatide binding and growth rate were retained for approximately 60 generations, after which the heat-shocked organisms reverted to their original glycolipid binding pattern (i.e., PE, Gg3, and Gg4) and growth rate . Such organisms could then be reexposed to heat, and the heat shock phenotype would be reestablished . After exposure of the organisms to brief heat shock, Western blotting of a surface extract of H . influenzae with anti-bovine-brain hsp-70 monoclonal antibody showed an increase in two protein bands at 82 and 60 kDa . This antibody was a potent inhibitor of the binding of heat-shocked H . influenzae to SGC and SGG but had no effect on PE, Gg3, or Gg4 binding in vitro . In contrast, an antibody against an H . influenzae PE-Gg3-Gg4-binding adhesin that was recently identified (J . Busse, E . Hartmann, and C . A . Lingwood, J . Infect . Dis . 175:77-83, 1996) selectively inhibited the organism's binding to PE and Gg3 . This indicates that cell surface hsp-70-related heat shock proteins can mediate H . influenzae attachment to sulfoglycolipids following heat shock . We suggest that such increased binding to sulfated glycolipids may be a response to fever following H . influenzae infection in humans.

Infect Immun, 1997 May, 65(5), 1710 - 5
MF59 adjuvant enhances antibody responses of infant baboons immunized with Haemophilus influenzae type b and Neisseria meningitidis group C oligosaccharide-CRM197 conjugate vaccine; Granoff DM et al.; The ability of the adjuvant MF59 to enhance the immunogenicity of polysaccharide-protein conjugate vaccines was investigated in infant baboons . MF59 consists of stable droplets (<250 nm) of the metabolizable oil squalene and two surfactants, polyoxyethylene sorbitan monooleate and sorbitan trioleate, in an oil-in-water emulsion . In humans, MF59 is well tolerated and enhances the immunogenicity of recombinant protein subunit or particle vaccines . Its effect on the immunogenicity of polysaccharide-protein conjugate vaccines is unknown . Baboons 1 to 4 months of age were immunized intramuscularly with Neisseria meningitidis group C and Haemophilus influenzae type b (Hib) oligosaccharide-CRM197 conjugate vaccines . The lyophilized vaccines were reconstituted with phosphate-buffered saline (PBS), Al(OH)3 (alum), or MF59 . Groups of five animals each were given three injections of the respective formulations, with one injection every 4 weeks . Four weeks after each immunization, the MF59 group had up to 7-fold-higher geometric mean anticapsular-antibody titers than the alum group and 5- to 10-fold-higher N . meningitidis group C bactericidal-antibody titers . Twenty-one weeks after the third immunization, the MF59 group still showed 5- to 10-fold-higher anticapsular-antibody titers . The antibody responses of the animals given the vaccines reconstituted with PBS were low at all times measured . Both the MF59 and alum groups, but not the PBS group, showed booster antibody responses to unconjugated Hib and N . meningitidis group C polysaccharides, results consistent with induction of memory B cells . Thus, MF59 may be useful for accelerating and augmenting immunity to polysaccharide-protein conjugate vaccines in infants.

Infect Immun, 1997 May, 65(5), 1695 - 700
Specific binding of Haemophilus influenzae to minor gangliosides of human respiratory epithelial cells; Fakih MG et al.; Gangliosides are sialylated glycosphingolipids that serve as receptors for various bacteria . To investigate endogenous gangliosides of human respiratory epithelial cells as potential receptors for Haemophilus influenzae, three strains, including nontypeable H . influenzae (NTHI) 1479, and isogenic fimbriated (f+) and nonfimbriated (f0) H . influenzae type b 770235, were 3H labeled and overlaid on two-dimensional thin-layer chromatography (TLC) plates containing either purified HEp-2 gangliosides or murine brain gangliosides . NTHI 1479 bound exclusively to two distinct minor ganglioside doublets, with mobilities near that of GM1 . These minor gangliosides comprised only 14.2 and 9.4% of the total, respectively . NTHI 1479 also bound to a distinct ganglioside of human macrophages whose chromatographic mobilities closely resemble those of one of the NTHI-binding gangliosides of HEp-2 cells . H . influenzae type b 770235 f+ and f0 each bound to a different minor HEp-2 ganglioside doublet, with proportionately weaker affinity for a major ganglioside doublet . Remarkably, none of the three strains bound to any murine brain gangliosides . Moreover, when 80 to 90% of sialic acid residues were enzymatically removed from HEp-2 gangliosides, NTHI 1479 binding was proportionately impaired, compared with untreated controls . Our findings support a role for specific gangliosides of specific cells as receptors for H . influenzae strains . Our findings further demonstrate that individual minor gangliosides possess unique biological properties.

J Clin Microbiol, 1997 May, 35(5), 1077 - 9
Comparison of four different sampling methods for detecting pharyngeal carriage of Streptococcus pneumoniae and Haemophilus influenzae in children; Rapola S et al.; Samples from 96 children with acute respiratory infection were obtained simultaneously with nasal, nasopharyngeal, and oropharyngeal swabs and by nasopharyngeal aspiration and were cultured on chocolate and blood agar plates . The rates of isolation of Streptococcus pneumoniae and Haemophilus influenzae detected by the four sampling methods were compared . Nasopharyngeal aspirates were optimal for the detection of both S . pneumoniae (isolation rate, 33%) and H . influenzae (isolation rate, 31%) . When a nasopharyngeal aspirate is not available, such as for healthy children or children with no obtainable secretions, the nasopharyngeal swab seems optimal for the detection of both S . pneumoniae and H . influenzae among children younger than 13 months of age . Among older children, similarly, the nasopharyngeal swab seems optimal for the detection of S . pneumoniae; however, for H . influenzae, the oropharyngeal swab seems optimal.

Lancet, 1997 Apr 26, 349(9060), 1197 - 202
Vaccine failures after primary immunisation with Haemophilus influenzae type-b conjugate vaccine without booster; Booy R et al.; BACKGROUND: Diseases of early childhood associated with Haemophilus influenzae type b (Hib) can now be prevented by vaccination . The rapid implementation of routine infant vaccination with Hib polysaccharide-tetanus protein conjugate (PRP-T) vaccine has allowed us to assess whether an accelerated 2, 3, and 4 month schedule can protect in the longer term without a booster dose and whether carrier priming influences protective efficacy . The degree of protection afforded by a catch-up programme with Hib oligosaccharide conjugate (HbOC) for older children was also assessed . METHODS: Paediatricians and microbiologists in the UK were asked to report all cases of invasive H influenzae infection in children who had received at least one dose of Hib-conjugate vaccine . Serum samples from convalescent children were obtained and the isolate was verified . Efficacy was estimated by comparing observed rates of Hib disease in those who had been vaccinated with rates predicted by age adjustment of disease rates from the prevaccine era . FINDINGS: Of 164 reports of invasive infection between Oct 1, 1992, and Oct 1, 1995, 43 were considered true vaccine failures . The estimated overall efficacy for three doses of PRP-T was 98.1% (95% CI 97.3-98.7%) . Efficacy in infants aged 5-11 months was 99.1%, 12-23 months 97.3%, and 24-35 months 94.7% . In infants aged 3-11 months, who received their first dose of PRP-T after tetanus toxoid vaccination, disease was unlikely from 1 week after one dose of PRP-T vaccine (88.6% protection in the second to fourth weeks {66.8-97.7%}) . The disease rate in vaccinated infants aged 2 months has declined year on year . In children aged 13 months to 2 years given HbOC, as a catch-up vaccine, the estimated efficacy was 94.0% (84.7-98.4%) . INTERPRETATION: A high degree of efficacy has been observed with PRP-T vaccine given as a three-dose schedule in infancy and with HbOC as a single dose in older children . Efficacy of PRP-T appears to be enhanced by carrier priming . Although with increasing age there was a small decline in efficacy of PRP-T, Hib disease is now close to being eliminated in the UK, and we suggest that a booster is not necessary in the second year of life.

Lancet, 1997 Apr 26, 349(9060), 1191 - 7
Randomised trial of Haemophilus influenzae type-b tetanus protein conjugate vaccine {corrected} for prevention of pneumonia and meningitis in Gambian infants; Mulholland K et al.; BACKGROUND: In developing countries, pneumonia and meningitis due to Haemophilus influenzae type b (Hib) are common in children under age 12 months and the mortality from meningitis is high . Protein-polysaccharide conjugate vaccines have brought Hib disease under control in industrialised countries . We did a double-blind randomised trial in The Gambia to assess the efficacy of a Hib conjugate vaccine for the prevention of meningitis, pneumonia, and other invasive diseases due to Hib . METHODS: Between March, 1993, and October, 1995, 42,848 infants were randomly allocated the conjugate vaccine Hib polysaccharide tetanus protein (PRP-T) mixed with diphtheria-tetanus-pertussis vaccine (DTP), or DTP alone at age 2 months, 3 months, and 4 months . Children who presented with signs of invasive Hib were investigated by blood culture and, where appropriate, by lumbar puncture, chest radiograph, or percutaneous lung aspirate . Children were followed up for between 5 and 36 months . FINDINGS: The median ages at which children received the study vaccine were 11 weeks, 18 weeks, and 24 weeks . 83% of children enrolled received all three doses of vaccine . 17 cases of culture-positive Hib pneumonia, 28 of Hib meningitis, and five of other forms of invasive Hib disease were detected amongst the study children . The efficacy of the vaccine for the prevention of all invasive disease after three doses was 95% (PRP-T vaccinees 1, controls 19 {95% CI 67-100}), for the prevention of Hib pneumonia after two or three doses, 100% (vaccinees 0, controls 10 {55-100}), and for the prevention of radiologically defined pneumonia at any time after enrollment, 21.1% (PRP-T vaccinees 198, controls 251 {4.6-34.9}) . INTERPRETATION: PRP-T conjugate Hib vaccine prevented most cases of meningitis and pneumonia due to Hib in Gambian infants . The reduction in the overall incidence of radiologically defined pneumonia in PRP-T vaccinees suggests that about 20% of episodes of pneumonia in young Gambian children are due to Hib . The introduction of Hib vaccines into developing countries should substantially reduce childhood mortality due to pneumonia and meningitis.

FEBS Lett, 1997 Apr 21, 407(1), 32 - 6
Scavengase p20: a novel family of bacterial antioxidant enzymes; Wan XY et al.; A novel antioxidant enzyme designated scavengase p20 was identified in various pathogenic bacteria through database searching for sequences strikingly homologous to a recently discovered Escherichia coli thiol peroxidase p20 . The direct biochemical evidence for the existence of scavengase p20 in Haemophilus influenzae, Streptococcus pneumoniae and Helicobacter pylori was provided by protein microsequencing and by in vitro assays for antioxidant activities . Overlapping genes encoding scavengase p20 and superoxide dismutase were recognized in H . pylori and their functional implications are discussed.

Cell, 1997 Apr 18, 89(2), 227 - 37
Molecular organization in site-specific recombination: the catalytic domain of bacteriophage HP1 integrase at 2.7 A resolution; Hickman AB et al.; HP1 integrase promotes site-specific recombination of the HP1 genome into that of Haemophilus influenzae . The isolated C-terminal domain (residues 165-337) of the protein interacts with the recombination site and contains the four catalytic residues conserved in the integrase family . This domain represents a novel fold consisting principally of well-packed alpha helices, a surface beta sheet, and an ordered 17-residue C-terminal tail . The conserved triad of basic residues and the active-site tyrosine are contributed by a single monomer and occupy fixed positions in a defined active-site cleft . Dimers are formed by mutual interactions of the tail of one monomer with an adjacent monomer; this orients active-site clefts antiparallel to each other.

Biochem J, 1997 Apr 15, 323 ( Pt 2), 365 - 70
Cloning, sequencing and expression of rat liver 3-phosphoglycerate dehydrogenase; Achouri Y et al.; Rat liver d-3-phosphoglycerate dehydrogenase was purified to homogeneity and digested with trypsin, and the sequences of two peptides were determined . This sequence information was used to screen a rat hepatoma cDNA library . Among 11 positive clones, two covered the whole coding sequence . The deduced amino acid sequence (533 residues; Mr 56493) shared closer similarity with Bacillus subtilis 3-phosphoglycerate dehydrogenase than with the enzymes from Escherichia coli, Haemophilus influenzae and Saccharomyces cerevisiae . In all cases the similarity was most apparent in the substrate- and NAD+-binding domains, and low or insignificant in the C-terminal domain . A corresponding 2.1 kb mRNA was present in rat tissues including kidney, brain and testis, whatever the dietary status, and also in livers of animals fed a protein-free, carbohydrate-rich diet, but not in livers of control rats, suggesting transcriptional regulation . The full-length rat 3-phosphoglycerate dehydrogenase was expressed in E . coli and purified . The recombinant enzyme and the protein purified from liver displayed hyperbolic kinetics with respect to 3-phosphoglycerate, NAD+ and NADH, but substrate inhibition by 3-phosphohydroxypyruvate was observed; this inhibition was antagonized by salts . Similar properties were observed with a truncated form of 3-phosphoglycerate dehydrogenase lacking the C-terminal domain, indicating that the latter is not implicated in substrate inhibition or in salt effects . By contrast with the bacterial enzyme, rat 3-phosphoglycerate dehydrogenase did not catalyse the reduction of 2-oxoglutarate, indicating that this enzyme is not involved in human D- or L-hydroxyglutaric aciduria.

Proc Natl Acad Sci U S A, 1997 Apr 15, 94(8), 4056 - 61
A diffusible cytotoxin of Haemophilus ducreyi; Cope LD et al.; Little is known about the virulence mechanisms employed by Haemophilus ducreyi in the production of genital ulcers . This Gram-negative bacterium previously has been shown to produce a soluble cytotoxic activity that kills HeLa and HEp-2 cells . We have now identified a cluster of three H . ducreyi genes that encode this cytotoxic activity . The predicted proteins encoded by these genes are most similar to the products of the Escherichia coli cdtABC genes that comprise the cytolethal distending toxin (CDT) of this enteric pathogen . Eleven of 12 H . ducreyi strains were shown to possess this gene cluster and culture supernatants from these strains readily killed HeLa cells . The culture supernatant from a single strain of H . ducreyi that lacked these genes was unable to kill HeLa cells . When the H . ducreyi cdtABC gene cluster was cloned into E . coli, culture supernatant from the recombinant E . coli clone killed HeLa cells . A monoclonal antibody that neutralized this soluble cytotoxic activity of H . ducreyi was shown to bind to the H . ducreyi cdtC gene product . This soluble H . ducreyi cytotoxin may play a role in the development or persistence of the ulcerative lesions characteristic of chancroid.

JAMA, 1997 Apr 9, 277(14), 1148 - 51
A national survey of immunization practices following allogeneic bone marrow transplantation; Henning KJ et al.; OBJECTIVES: To describe the frequency and patterns of use of routine childhood and hepatitis B, pneumococcal, influenza, and meningococcal vaccines following allogeneic bone marrow transplantation (BMT) . DESIGN, SETTING, AND PARTICIPANTS: Survey of all US transplantation centers participating in the National Marrow Donor Program (NMDP) during 1994 . MAIN OUTCOME MEASURES: Use, timing, and total doses of selected vaccines given to patients younger than 7 years and patients aged 7 years or older following allogeneic BMT . RESULTS: Of 66 centers associated with the NMDP, 45 (68%) responded . A total of 97% of centers performing transplants on patients younger than 7 years and 88% of centers performing transplants on patients aged 7 years or older gave either the diphtheria-tetanus vaccine or the diphtheria-tetanus-pertussis vaccine compared with 77% and 58% usage, respectively, of Haemophilus influenza type b conjugate vaccine (P=.03 and .003, respectively) . Centers were more likely to administer inactivated poliovirus and measles-mumps-rubella vaccines to patients younger than 7 years than to the older age group (94% vs 73% for poliovirus, P=.02; and 94% vs 70% for measles-mumps-rubella, P=.01) . About one half of centers routinely administer hepatitis B vaccine and approximately three quarters immunize with pneumococcal and influenza vaccines . Few programs, regardless of age of bone marrow recipient, use multiple vaccine (> or =2) doses . The number of schedules reported for specific vaccines varied widely (3-11 schedules per vaccine) . CONCLUSIONS: Despite convincing evidence that patients lose protective antibodies to vaccine-preventable diseases following allogeneic BMT and accumulating data showing the safety and efficacy of many vaccines after BMT, vaccines are underutilized and schedules vary widely at US transplant centers . National guidelines for optimal doses and timing of vaccines after BMT are warranted.

Eur J Epidemiol, 1997 Apr, 13(3), 317 - 21
Pattern of bacterial meningitis in Italy, 1994; Salmaso S et al.; During 1994, 603 cases of bacterial meningitis were reported in Italy . Seventy-five percent of cases with determined etiology was due to three agents: Neisseria meningitidis (33.4%), Streptococcus pneumoniae (23.4%) and Haemophilus influenzae (18.6%) . The majority of cases due to N . meningitidis and H . influenzae occurred in subjects below five years of age (35.7% and 84.8%, respectively) while S . pneumoniae accounted for 52.8% of meningitis cases in subjects older than 44 year of age . The estimated incidence of N . meningitidis on the national population in 1994 was 0.27 per 100,000 . Serogroup B accounted for 62.5% of the serotyped isolates, group C for 23.1%, group A for 7.2%, group W135 for 3.6%, group Y for 1.8% . All tested meningococcal strains were susceptible to penicillin as well as to rifampin . Incidence of meningococcal meningitis in 1994 has been low suggesting that its relative importance compared to other bacteria causing meningitis is likely to change in the future . Therefore, extended surveillance on bacterial meningitis by other etiological agents has to be maintained and implemented in order to undertake the appropriate control measures and evaluate their effect.

Vet Microbiol, 1997 Apr, 55(1-4), 247 - 57
Porcine reproductive and respiratory syndrome virus (PRRSv) interaction with Haemophilus parasuis; Solano GI et al.; The interaction of bacteria and virus has been well demonstrated in the pathogenesis of respiratory disease in swine . The interaction between porcine respiratory and reproductive syndrome virus (PRRSv) and Haemophilus parasuis has not been studied . We initiated studies to evaluate a possible effect of the PRRSv on the pathogenesis of polyserositis caused by H . parasuis . A group of 30 three week old piglets were distributed in 4 groups . Group I (10 pigs) was inoculated with PRRSv and H . parasuis . Group II (10 pigs) was inoculated with H . parasuis alone . Group III (5 pigs) was inoculated with virus alone and group IV (5 pigs) was inoculated with culture media . Lesions consisted of a severe fibrinous polyserositis affecting 7 of 10 animals in group II and a mild fibrinous pleuritis in 1 of 10 animals of group I . Three of ten animals dually infected with the two agents died during the course of the study . These animals had pulmonary congestion and focal lung hemorrhages . No other animals died from other groups . Group III and IV had no macroscopic lesions . Microscopically group III had interstitial pneumonia . Immunomodulating virus effect may explain the differences in terms of lesions severity between groups I and II . Septic shock was suspected as cause of sudden death.

Vet Microbiol, 1997 Apr, 55(1-4), 187 - 96
General overview of PRRSV: a perspective from the United States; Zimmerman JJ et al.; Four years after the report of its discovery, porcine reproductive and respiratory syndrome virus (PRRSV) continues to challenge swine producers, veterinary practitioners, and animal health researchers in the United States . The prevalence of infection is high--60% to 80% of herds is a reasonable estimate--but the clinical effects of infection vary widely among farms . In many herds, infection is unapparent and productivity seemingly unaffected . Some infected herds report occasional respiratory disease outbreaks in young pigs, or periodic outbreaks of reproductive disease, and a few herds experience severe, chronic disease problems, particularly in young pigs . In these herds, secondary infections with viral or bacterial pathogens, particularly Salmonella choleraesuis, Streptococcus suis, or Haemophilus parasuis typically occur concurrently with PRRSV infections . Understanding why some herds undergo devastating episodes of clinical disease and others show no apparent effects is central to solving the problem of clinical PRRS for swine producers . Understanding the ecology and epidemiology of PRRSV is the key to preventing and controlling PRRSV in the future . The objective of this article is to review recent developments in these areas.

Emerg Infect Dis, 1997 Apr-Jun, 3(2), 179 - 82
Invasive Haemophilus influenzae type b disease in elderly nursing home residents: two related cases; Heath TC et al.; We investigated two fatal cases of invasive Haemophilus influenzae type b (Hib) infection in a community nursing home in western Sydney, Australia . Two elderly women had lived in the same room, and the onset of their illness was 5 days apart . Hib isolates from blood cultures showed identical profiles by pulsed field gel electrophoresis . These findings suggest that Hib infection was transmitted within this nursing home . Serious Hib disease may be underrecognized in this setting . Continued surveillance and serotyping of invasive H . influenzae disease is essential for identifying groups at increasing risk that may benefit from immunization against Hib.

Arch Pediatr, 1997 Apr, 4(4), 325 - 30
{Otorrhea on transtympanic aerator}; Essaadi M et al.; BACKGROUND: Purulent otorrhea is the most common complication of tympanostomy tubes . POPULATION AND METHODS: Results of culture of otorrhea in 33 consecutive cases were compared to two similar studies performed 4 and 8 years ago in the same institution . RESULTS: The most frequent organisms were Pseudomonas aeruginosa, Haemophilus influenzae, Staphylococcus aureus and Streptococcus pneumoniae (45, 24, 21 and 15% of the cases, respectively) . Such flora resembles that of external otitis and chronic otitis media (P aeruginosa and S pneumoniae) . CONCLUSION: Sensitivity of the organisms encountered in these otorrheas favors the use of topical drops rather than oral antibiotics as the first choice of treatment.

Vaccine, 1997 Apr-May, 15(6-7), 775 - 81
Immunogenicity and safety of Haemophilus influenzae type b polysaccharide-Neisseria meningitidis conjugate vaccine in 7.5 micrograms liquid formulation: a comparison of three lots with the 15.0 micrograms lyophilized formulation . Study Group for 7.5 micrograms Liquid PedvaxHIB; Mendelman PM et al.; We conducted a multicenter, single-blind, randomized comparisons of the immunogenicity and safety of three manufacturing-scale lots of 7.5 micrograms liquid Haemophilus influenzae type b polysaccharide- Neisseria meningitidis conjugate vaccine (PRP-OMPC) and a single lot of 15.0 micrograms lyophilized PRP OMPC . A total of 908 infants were entered into the study . Each infant received two primary injections intramuscularly 2 months apart beginning at age 2-6 months and a booster injection at 12-15 months . Blood samples for serology were obtained before each injection and 1 month after the second and the booster dose . Immune responses were measured by radioimmunoassay . Approximately 80% of the infants achieved a titer > 1.0 micrograms ml-1 after the second primary dose of all four lots tested: the geometric mean titer (GMT) was ca 3 micrograms ml-1 for each vaccine group . After the booster dose, more than 90% of infants from each vaccine group had a titer > 1.0 microgram ml-1;GMTs ranged from 8 to 10 micrograms ml-1 . No serious vaccine-associated adverse reactions were reported . Thus the 7.5 liquid PRP OMPC vaccine was at least as immunogenic and well tolerated as the 15.0 micrograms lyophilized vaccine.

Int J Epidemiol, 1997 Apr, 26(2), 443 - 50
Protective effect of breastfeeding on invasive Haemophilus influenzae infection: a case-control study in Swedish preschool children; Silfverdal SA et al.; BACKGROUND: In Orebro County a 2.5-fold increase in the incidence of Haemophilus influenzae (HI) meningitis was found between 1970 and 1980, an observation that initiated the present study . MATERIALS AND METHODS: In order to search for associations between morbidity in invasive HI infection and possible risk factors, a case-control study was conducted over a 6-year period from 1987 to 1992, before general Hib vaccination was introduced in Sweden . Fifty-four cases with invasive HI infection 139 matched controls were studied for possible risk factors such as day-care outside the home, short duration of breastfeeding, passive smoking, low socioeconomic level of the household, many siblings in the family, allergy, frequent, infections, repeated antibiotic treatments and immunoglobulin deficiency . RESULTS: Multivariate analysis showed a significant association between invasive HI infection and two independent factors, i.e . short duration (< 13 weeks) of exclusive breastfeeding, odds ratio (OR) 3.79 (95% confidence interval {CI} 1.6-8.8) and history of frequent infections, OR 4.49 (95% CI : 1.0-21.0) . For the age at onset 12 months or older, the associations were stronger, OR 7.79 (95% CI : 2.4-26.6) and 5.86 (95% CI : 1.1-30.6), respectively . When breastfeeding duration in weeks was analysed as a continuous variable the OR was 0.95 (95% CI : 0.92-0.99), indicating a decreased risk with each additional week . Increased OR were observed for other risk factors as well but not of the magnitude found for short duration of breastfeeding . DISCUSSION: The association of decreased risk for invasive HI infection and long duration of breastfeeding was persisting beyond the period of breastfeeding itself . This finding supports the hypothesis of a long-lasting protective effect of breastfeeding on the risk for invasive HI infection . CONCLUSION: A decreased risk for invasive HI infection with long duration of breastfeeding was found . Our results do have implications for strategies in breastfeeding promotion, especially in countries where Hib vaccination is too costly and not yet implemented.

Arch Dis Child, 1997 Apr, 76(4), 334 - 6
Respiratory morbidity from lymphocytic interstitial pneumonitis (LIP) in vertically acquired HIV infection; Sharland M et al.; The aim of the study was to define the respiratory morbidity caused by lymphocytic interstitial pneumonitis (LIP) in children with vertically acquired HIV infection . A retrospective case note review was performed on 95 children attending three London hospitals . Clinical and radiological evidence of LIP, acute lower respiratory tract infections, and chronic lung disease was obtained using a structured protocol . A diagnosis of LIP had been made in 33%, and an acute admission due to acute lower respiratory tract infection had occurred in 42% of all children (despite 99% taking regular cotrimoxazole prophylaxis) . Admission rates because of acute lower respiratory tract infection were significantly higher in the LIP group (0.38 admissions/child year) than in the non-LIP group (0.17 admissions/child year) (p = 0.0002) . Encapsulated bacteria (Streptococcus pneumoniae, Haemophilus influenzae) were most frequently isolated . Improved methods of prevention of acute lower respiratory tract infection may help to reduce the severe respiratory morbidity seen in children with LIP and HIV infection.

Diagn Microbiol Infect Dis, 1997 Apr, 27(4), 139 - 45
Influence of pH on the antimicrobial activity of clarithromycin and 14-hydroxyclarithromycin against Haemophilus influenzae using an in vitro pharmacodynamic model; Garrison MW et al.; Clarithromycin activity can be influenced by the pH of the surrounding environment . Evidence supports a reduced pH of middle ear fluid (MEF) and lung tissues in patients with otitis media and pneumonia, respectively . To evaluate the influence of pH on clarithromycin activity, an in vitro pharmacodynamic chamber model (PDCM) was used to generate bacterial time-kill curves for clarithromycin and a 2:1 ratio of clarithromycin and 14-hydroxyclarithromycin (HC) against Haemophilus influenzae at three different pH values: 7.2, 6.8, 6.4 . Concentrations observed in MEF and lung tissues were simulated for clarithromycin alone and clarithromycin plus HC . Differences in activity at each pH were identified by comparing initial kill curve slopes and total log reduction . Experiments with amoxicillin-clavulanate were conducted as a reference . In simulated MEF regimens at pH 7.2, activity of clarithromycin alone improved by adding HC (additional 2 log10 reduction at 8 h); however, at pH values of 6.8 and 6.4, kill curves resembled growth controls . In simulated lung regimens, differences between clarithromycin alone and clarithromycin plus HC were insignificant; both produced a 2 log10 reduction at pH 7.2, and activity dramatically dropped to < 0.4 log10 as pH declined . In contrast, amoxicillin-clavulanate consistently produced a 3 log10 reduction over each pH value with more rapid initial kill relative to all clarithromycin regimens . These findings suggest the activity of clarithromycin against H . influenzae may be significantly compromised in respiratory tract infections involving a reduced pH . Trials with emphasis on clinical outcomes analysis will assist further in determining the significance of these experimental findings.

Zentralbl Veterinarmed B, 1997 Apr, 44(2), 73 - 86
Characterization of a large transferrin-binding protein from Actinobacillus pleuropneumoniae serotype 7; Wilke M et al.; The binding of transferrin at the surface of Actinobacillus pleuropneumoniae (A . pp.) is mediated by two proteins of approximately 60 and 100 kDa . The 60 kDa protein has been shown to be highly divergent among different serotypes and to induce a serotype-specific protective immune response . In this study we have characterized the 100 kDa transferrin-binding protein of A . pp . serotype 7 and designated it as TfbB . The tfbB gene was found to be located immediately downstream of the tfbA gene . It was cloned and sequenced, and antibodies raised against the isolated recombinant protein detected, with a constant intensity, a 100 kDa protein in A . pp . serotypes 2, 4, 6, 7, 8, 9, 10 and 11, and a polypeptide of approximately 103 kDa in serotypes 1, 3, 5A and 12 . In addition, comparative analysis of the deduced amino acid sequence showed more than 40% identity with the large transferrin-binding proteins of Neisseria meningitidis and Haemophilus influenzae . The TfbB protein was expressed in E . coli outer membranes in a conformation eliciting porcine transferrin-specific binding activity . Sera of pigs immunized with these TfbB-containing E . coli membranes recognized functional membrane-associated TfbB protein whereas no such reaction was observed upon immunization with isolated recombinant TfbB protein . A preliminary animal experiment showed that TfbB-containing outer membrane preparations from recombinant E . coli can reduce significantly the mortality of an A.pp . infection with the homologous strain.

Dtsch Tierarztl Wochenschr, 1997 Apr, 104(4), 150 - 3
Association of Pasteurella haemolytica, Pasteurella multocida and Haemophilus somnus with pneumonia in calves; Haziroglu R et al.; Pathological and bacteriological observations were made on 100 (0.98%) pneumonic lungs of 10140 slaughtered beef calves during March 1995-June 1996 period . Gross lesions were mainly lobular and occasionally lobar pattern and, were frequently observed in the pars cranialis of lobus cranialis dexter . In histological examination, proliferative-exudative pneumonia was observed in 79 cases, and proliferative pneumonia alone in 21 cases . In bacteriological examination, Pasteurella haemolytica . Pasteurella multocida and Haemophilus somnus were isolated from 42.8 and 10 of pneumonic lungs respectively . In 7 cases, P . haemolytica and H . somnus were isolated from the same sample P . haemolytica and P . multocida were also found in the same sample in 2 cases . There was a close relation among these organisms and exudative inflammation (P < 0.01).

J Antimicrob Chemother, 1997 Apr, 39(4), 499 - 508
Atypical pneumonia in the Nordic countries: aetiology and clinical results of a trial comparing fleroxacin and doxycycline . Nordic Atypical Pneumonia Study Group; Ragnar Norrby S; Community-aquired pneumonia caused by atypical bacteria or viruses was studied in a double-blind trial comparing fleroxacin 400 mg od and doxycycline 100 mg bd for 10 days . The aetiology was confirmed in 258 of 411 cases (66%), of which 133 were caused by Mycoplasma spp., Chlamydia spp . or Legionella spp.; 30 patients had viral infection, nine had pneumococcal or Haemophilus influenzae infection and 93 had mixed aetiology . In intention-to-treat analyses clinical response rates in fleroxacin-treated patients were 86% (157/182) and 75% (137/182) 2-8 days and 3-5 weeks after therapy, respectively . Corresponding results with doxycycline were 93% (177/191) and 85% (162/190), respectively . Differences between treatments seemed to be due to the lower activity of fleroxacin compared with doxycycline against mycoplasma and pneumococci . Drug-related adverse events were reported in 39% of 204 fleroxacin patients and in 34% of 207 doxycycline patients . The null hypothesis that fleroxacin was <15% inferior to doxycycline was accepted at early follow-up but rejected at later review.

J Paediatr Child Health, 1997 Apr, 33(2), 138 - 41
The immunogenicity of Haemophilus influenzae: meningococcal protein conjugate vaccine in Polynesian and non-Polynesian New Zealand infants; Ameratunga SN et al.; OBJECTIVE: To examine the comparative immunogenicity of the Haemophilus influenzae type b-meningococcal protein (PRP-OMP) conjugate vaccine in Polynesian and non-Polynesian New Zealand infants . METHODOLOGY: Fifty-six Polynesian and 53 non-Polynesian infants aged 2-7 months recruited from primary health care settings in Auckland received a two-dose primary series of PRP-OMP . A sub-sample of 83 participants received a booster dose of PRP-OMP at 12-16 months of age . Anti-PRP antibody concentrations were measured in pre- and post-vaccination blood samples . RESULTS: Antibody responses consistent with long-term protection (> or = 1.00 microgram/mL) were observed in 72, 85 and 95% of children following the first, second and booster doses . CONCLUSIONS: Despite differences in disease epidemiology, PRP-OMP was highly immunogenic in Polynesian and non-Polynesian infants.

FEMS Immunol Med Microbiol, 1997 Apr, 17(4), 235 - 42
Distribution, cloning, characterisation and mutagenesis of sodC, the gene encoding copper/zinc superoxide dismutase, a potential determinant of virulence, in Haemophilus ducreyi; Langford PR et al.; The sodC gene encoding the periplasmic enzyme copper/zinc superoxide dismutase (CuZnSOD) has been cloned from Haemophilus ducreyi, the causative agent of the genital ulcer disease, chancroid . Examination of a collection of diverse strains indicates that it is present throughout the species . Cloned sodC has been expressed in E . coli and shown to encode active enzyme . Insertional mutagenesis was used to construct a non-functional version of the gene . This has been transferred into the chromosome of the parent H . ducreyi strain by electroporation and homologous recombination, in preparation for studies of the role of this enzyme in the interactive biology of the organism with its host, perhaps in protecting bacteria from superoxide radicals and their reactive progeny generated by neutrophils in the context of host defence.

Microbiology, 1997 Apr, 143 ( Pt 4), 1423 - 31
Differences in genetic diversity of nonecapsulated Haemophilus influenzae from various diseases; van Alphen L et al.; Genetic relationships among 80 isolates of nonencapsulated Haemophilus influenzae recovered from different disease types were determined by multilocus enzyme electrophoresis (MEE) at 13 enzyme loci in an attempt to assess the association between multilocus genotype and disease . The isolates were obtained from 15 patients with meningitis, 10 with otitis media, 19 with chronic bronchitis, 20 with cystic fibrosis, and 16 were obtained from healthy carriers . The 80 isolates were assigned to 69 electrophoretic types (ETs) falling into 5 groups . Isolates from each disease entity were represented by a variety of genotypes; however, cluster analysis from a matrix of genetic distances between ETs revealed that the ETs of the otitis media and meningitis isolates were all clustered within a genetic distance of 0.55 (group I) . In addition, no genotypes were shared between H . influenzae carrier isolates and isolates from cases of disease, H . influenzae isolates from healthy individuals were distributed significantly differently from those from chronic bronchitis meningitis and otitis media patients . The genetic diversity (H) of carrier strains was greatest, although not statistically different from that of isolates from patients with disease . It was concluded that the genetic distribution of acute disease isolates is not random over the five ET groups, although the genetic diversity within the groups is not different . The effect of bacterial persistence in the host on the genetic diversity of H . influenzae is discussed.

Microbiology, 1997 Apr, 143 ( Pt 4), 1287 - 97
Structure and gene-polypeptide relationships of the region encoding glycerol diffusion facilitator (glpF) and glycerol kinase (glpK) of Pseudomonas aeruginosa; Schweizer HP et al.; The glycerol facilitator is one of the few known examples of bacterial solute transport proteins that catalyse facilitated diffusion across the cytoplasmic membrane . A second protein, glycerol kinase, is involved in entry of external glycerol into cellular metabolism by trapping glycerol in the cytoplasm as sn-glycerol 3-phosphate . Evidence is presented that glycerol transport in Pseudomonas aeruginosa is mediated by a similar transport system . The genes encoding the glycerol facilitator, glpF, and glycerol kinase, glpK, were isolated on a 4.5 kb EcoRI fragment from a chromosomal mini-library by functional complementation of an Escherichia coli glpK mutant after establishing a map of the chromosomal glpFK region with the help of a PCR-amplified glpK segment . The nucleotide sequence revealed that glpF is the promoter-proximal gene of the glpFK operon . The glycerol facilitator and glycerol kinase were identified in a T7 expression system as proteins with apparent molecular masses of 25 and 56 kDa, respectively . The identities of the glycerol facilitator and glycerol kinase amino acid sequences with their counterparts from Escherichia coli were 70 and 81%, respectively; this similarity extended to two homologues in the genome sequence of Haemophilus influenzae . A chromosomal delta glpFK mutant was isolated by gene replacement . This mutant no longer transported glycerol and could no longer utilize it as sole carbon and energy source . Two ORFs, orfX and orfY, encoding a putative regulatory protein and a carbohydrate kinase of unknown function, were located upstream of the glpFK operon.

Eur J Pediatr, 1997 Apr, 156(4), 288 - 91
A retrospective epidemiological study of bacterial meningitis in an urban area in Belgium; van Hoeck KJ et al.; In order to obtain epidemiological data on the incidence of bacterial meningitis (BM) before the systematic introduction of vaccination against Haemophilus influenzae type b, a retrospective study of 124 children with proven BM was performed in an urban area in Belgium . N . meningitidis was the most prevalent cause, followed by H . influenzae and S . pneumoniae . Over a period of 6 years the incidence of BM increased ten fold, mainly due to an increase in N . meningitidis . The median age of the children with BM was 17 months and 35% of those with H . influenzae were younger than 1 year . Significant risk factors for BM as a whole were: age under 1 year, male gender, non-Caucasian descent and winter time . These findings may have implications for future vaccination policy in Belgium . CONCLUSION: Future vaccination schemes in Belgium should take into account than N . meningitis was the prevalent cause of bacterial meningitis and that certain factors increase the risk for developing bacterial meningitis.

Eur J Pediatr, 1997 Apr, 156(4), 282 - 7
Haemophilus influenzae b-vaccination: the urgency for timely vaccination; von Kries R et al.; In Germany the annual number of systemic Haemophilus influenzae cases in unvaccinated children aged 3-60 months has recently been exceeded by the number of cases in children vaccinated at least once with the PRP-D, HbOC or OMP vaccines, which until 1995 have almost exclusively been used for H . influenzae b (Hib) vaccination . Most of the vaccinated children however could already have had more vaccinations at onset of disease . How much does an age-related suboptimal vaccination status increase the risk for systemic H . influenzae infections? A case control study was performed in West Germany . Cases with systemic H . influenzae infections were ascertained between 7/92 and 8/ 94 with an ongoing active hospital surveillance programme . Six age-matched population controls per case were recruited at random . Only vaccinated cases and controls were included in the study . The main exposure analysed in this study was suboptimal vaccination at censoring; for censoring ages (age at disease onset in cases and corresponding age in matched controls) > 6 months: one vaccination in 1st year only; > 18 months: two (three for combined vaccines with Hib + DT or DPT in one syringe) vaccinations in the 1st year of life but no booster vaccination . Suboptimal vaccination for age increased the risk for systemic H . influenzae infections by a factor of 4.74 (95%-CI 2.17-10.34) . Following adjustment for confounders the odds ratio was 4.39 (95%-CI 1.74-11.07) . Subgroup analyses showed that this risk was not related to the type of vaccine used . The risk for "no booster vaccination" in children aged > 18 months appeared even greater than the risk associated with one vaccination in the 1st year only . CONCLUSIONS: On schedule and complete Hib vaccinations are essential for an optimal effectiveness of Hib vaccination programmes . Booster vaccinations between 12 and 18 months are important if the PRP-D, HbOC and OMP vaccines are used for primary vaccination.

Infect Immun, 1997 Apr, 65(4), 1468 - 74
Nonencapsulated Haemophilus influenzae in Aboriginal infants with otitis media: prolonged carriage of P2 porin variants and evidence for horizontal P2 gene transfer; Smith-Vaughan HC et al.; Aboriginal infants in the Northern Territory of Australia experience recurrent otitis media from an early age . Nonencapsulated Haemophilus influenzae (NCHi) colonization of the nasopharynx initially occurs within weeks of birth, persists throughout infancy and most of childhood, and contributes to otitis media . We established previously that the high carriage rates of NCHi in these infants result from concurrent and successive colonization with multiple strains, with sequential elimination of dominant strains . We have now sequenced loops 4, 5, and 6 of the NCHi P2 porin gene and characterized several strains with prolonged carriage times . Furthermore, despite a wide diversity of P2 gene sequences, we have four examples of P2 gene identity for strains with different genetic backgrounds as characterized by PCR ribotyping and randomly amplified polymorphic DNA typing, which leads us to suggest that the P2 gene has been transferred between strains . We also discuss the possibility that the paradoxical observation of cocolonization and prolonged carriage of P2-identical strains is related to immune suppression or tolerance in the host.

Infect Immun, 1997 Apr, 65(4), 1377 - 86
Identification of the ADP-L-glycero-D-manno-heptose-6-epimerase (rfaD) and heptosyltransferase II (rfaF) biosynthesis genes from nontypeable Haemophilus influenzae 2019; Nichols WA et al.; Haemophilus influenzae is an important human pathogen . The lipooligosaccharide (LOS) of H . influenzae has been implicated as a virulence determinant . To better understand the assembly of LOS in nontypeable H . influenzae (NtHi), we have cloned and characterized the rfaD and rfaF genes of NtHi 2019, which encode the ADP-L-glycero-D-manno-heptose-6-epimerase and heptosyltransferase II enzymes, respectively . This cloning was accomplished by the complementation of Salmonella typhimurium lipopolysaccharide (LPS) biosynthesis gene mutants . These deep rough mutants are novobiocin susceptible until complemented with the appropriate gene . In this manner, we are able to use novobiocin resistance to select for specific NtHi LOS inner core biosynthesis genes . Such a screening system yielded a plasmid with a 4.8-kb insert . This plasmid was able to complement both rfaD and rfaF mutants of S . typhimurium . The LPS of these complemented strains appeared identical to the wild-type Salmonella LPS . The genes encoding the rfaD and rfaF genes from NtHi 2019 were sequenced and found to be similar to the analogous genes from S . typhimurium and Escherichia coli . The rfaD gene encodes a polypeptide of 35 kDa and the rfaF encodes a protein of 39 kDa, as demonstrated by in vitro transcription-translation studies . Isogenic mutants which demonstrated truncated LOS consistent with inner core biosynthesis mutants were constructed in the NtHi strain 2019 . Primer extension analysis demonstrated the presence of a strong promoter upstream of rfaD but suggested only a very weak promoter upstream of rfaF . Complementation studies, however, suggest that the rfaF gene does have an independent promoter . Mass spectrometric analysis shows that the LOS molecules expressed by H . influenzae rfaD and rfaF mutant strains have identical molecular masses . Additional studies verified that in the rfaD mutant strain, D-glycero-D-manno-heptose is added to the LOS molecule in place of the usual L-glycero-D-manno-heptose . Finally, the genetic organizations of the inner core biosynthesis genes of S . typhimurium, E . coli, and several strains of H . influenzae were examined, and substantial differences were uncovered.

Infect Immun, 1997 Apr, 65(4), 1351 - 6
Molecular variation in the major outer membrane protein P5 gene of nonencapsulated Haemophilus influenzae during chronic infections; Duim B et al.; During the course of persistent infections by nonencapsulated Haemophilus influenzae in patients with chronic bronchitis, the major outer membrane protein (MOMP) P5 varies in molecular weight . The nature of this variability was determined by DNA sequence analysis of the P5 gene from five different H . influenzae strains and their seven MOMP P5 variants which were isolated from patients with chronic infections of the lower respiratory tract . Analysis of the P5 sequence data from the different strains revealed four well-defined, heterogeneous regions . These regions of variable sequence appeared to correspond to the regions of the gene encoding the putative surface-exposed loops of MOMP P5 . The MOMP P5 variants with alterations in MOMP P5 were shown to result from DNA point mutations and codon deletions . In addition, in three variants derived sequentially from one H . influenzae strain, a frameshift mutation resulted in the formation of a stop codon in the region encoding the signal sequence of the MOMP P5 gene . Strikingly, all nucleotide substitutions in the MOMP P5 loop regions of variants were nonsynonymous, suggesting that variants with alterated amino acid compositions of the surface-exposed parts of MOMP P5 obtained a selective advantage during persistence of the infection by nonencapsulated H . influenzae in chronic bronchitis patients.

Pediatr Infect Dis J, 1997 Apr, 16(4 Suppl), S97 - 102
Combination vaccines consisting of acellular pertussis vaccines; Edwards KM et al.; Combination vaccines consisting of multiple vaccine antigens delivered in a single injection simplify vaccine administration . Combining multiple antigens into one injection, however, presupposes that the administration of multiple vaccines in combination will not reduce the safety and immunogenicity of the component vaccines . New generation vaccines seek to combine many more antigens from multiple, different pathogens, making them difficult to study in controlled, double blind, randomized clinical trials because of the number of study arms required for complete evaluation . Methods to simplify studies of combination vaccines include building on the framework of earlier studies to calculate sample size and reduce the number of control arms, standardizing serologic assays and assessment of adverse reactions and determining serologic correlates of protection to minimize the need for multiple efficacy studies and to facilitate evaluation of immunogenicity studies . Preliminary data indicate that immune responses to combination vaccines including diphtheria-tetanus-acellular pertussis, hepatitis B and inactivated poliovirus are comparable with those seen when the vaccines are administered separately . In contrast studies of combination vaccines that include Haemophilus influenzae type b Hib antigens show a diminished Hib antibody response, although the clinical relevance of this lowered antibody response has not yet been determined . Numerous small safety studies of combined vaccines have not found evidence of increased adverse reactions.

Pediatr Infect Dis J, 1997 Apr, 16(4), 449 - 56
Causative pathogens, antibiotic resistance and therapeutic considerations in acute otitis media; Block SL; Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis are the most frequently isolated pathogens in patients with acute otitis media (AOM) . Other potential causative pathogens include Streptococcus pyogenes in older children and Chlamydia pneumoniae in younger children . The recent emergence of penicillin-resistant S . pneumoniae and the increasing frequency of beta-lactamase-producing strains of M . catarrhalis and H . influenzae are creating concerns regarding the use of amoxicillin as traditional first line empiric therapy for AOM in younger children . Both the in vitro antibiotic activity against these more resistant causative pathogens and the antibiotic concentrations achieved in middle ear fluid must be considered when selecting antibiotics for treatment of refractory AOM . The newer macrolides, azithromycin and clarithromycin, provide reasonable in vitro coverage against penicillin-resistant S . pneumoniae and beta-lactamase-producing H . influenzae, although azithromycin is more active against the latter . Both drugs also achieve notably higher, sustained concentrations in middle ear fluid than do beta-lactam antibiotics . Thus the newer macrolides represent important new rational alternatives for the management of AOM.

Pediatr Infect Dis J, 1997 Apr, 16(4), 432 - 7
Microbiologic activity of the newer macrolide antibiotics; McCracken GH Jr; In vitro susceptibility testing has demonstrated good activity of the azalide azithromycin and the macrolide clarithromycin against Gram-positive and -negative pathogens as well as atypical organisms involved in the etiology of upper and lower respiratory tract infections . One difference between these drugs in terms of their antimicrobial spectrum is the activity of azithromycin against Haemophilus influenzae . This organism is 2 to 8 times more susceptible in vitro to azithromycin than to clarithromycin or to erythromycin, the prototypical macrolide antibiotic . A principal concern in the management of respiratory tract infections today is the emergence of penicillin-resistant strains of Streptococcus pneumoniae . Both azithromycin and clarithromycin are active against penicillin-susceptible S . pneumoniae, although the activity of azithromycin is somewhat less than that of erythromycin and clarithromycin . Results of susceptibility testing of resistant organisms have varied among centers; in some areas all of the intermediately and some of the highly penicillin-resistant S . pneumoniae isolates are susceptible to the newer macrolides, whereas in other areas they are not . High tissue antibiotic concentrations achieved with these drugs may contribute to their effectiveness against some of the resistant S . pneumoniae isolates.

Pediatr Infect Dis J, 1997 Apr, 16(4), 386 - 90
Microbiology of recurrent parotitis; Giglio MS et al.; BACKGROUND: Infantile chronic recurrent parotitis (ICRP) is characterized by episodes of recurrent swelling of the parotid gland with decreased salivary flow and purulent secretion . The etiology of this little unknown clinical condition has been attributed to multiple causes such as canalicular system malformations, ascending bacterial infection, hyposialia, parotitis sequelae, viral infections and immunologic disorders, among others . METHODS: We studied the types (with counts) of microorganisms involved in ICRP . Saliva samples were obtained from 56 patients and 20 controls, inoculated onto enriched media and incubated under aerobic and anaerobic conditions . Antimicrobial susceptibility and serotyping of the isolated organisms isolated were performed . RESULTS: Of 57 saliva samples from ICRP patients, 52 (91%) were culture-positive . The most frequently isolated microorganisms were Streptococcus pneumoniae and Haemophilus influenzae . Thirteen of twenty (65%) samples were also culture-positive, mostly for viridans streptococci . However, colony counts were lower than in clinical samples (P < 0.004) . Approximately one-third of S . pneumoniae strains resistant or moderately resistant to penicillin, and all H . influenzae strains were susceptible to all of the antimicrobials tested . CONCLUSIONS: S . pneumoniae or H . influenzae were isolated in high concentrations in IRCP cases but not in controls, suggesting that these microorganisms may have a role in the development of this clinical entity . Quantitative cultures are very important in assessment of the pathogenic role of these microorganisms in patients but not in controls.

J Bacteriol, 1997 Apr, 179(7), 2267 - 73
aarC, an essential gene involved in density-dependent regulation of the 2'-N-acetyltransferase in Providencia stuartii; Rather PN et al.; The 2'-N-acetyltransferase {AAC(2')-Ia} in Providencia stuartii has a dual function where it is involved in the acetylation of peptidoglycan and certain aminoglycosides . A search for negative regulators of the aac(2')-Ia gene has resulted in the identification of aarC . A missense allele (aarC1) resulted in an 8.9-fold increase in beta-galactosidase accumulation from an aac(2')-lacZ transcriptional fusion . Northern blot analysis demonstrated an increase in aac(2')-Ia mRNA accumulation that was specific to cells at high density . In addition, the aarC1 allele also resulted in a substantial increase in the expression of aarP, a transcriptional activator of the aac(2')-Ia gene . The wild-type aarC gene was isolated by complementation and encodes a predicted protein of 365 amino acids with a molecular mass of 39,815 Da . The predicted AarC protein exhibited 88% amino acid homology to the previously identified GcpE protein of Escherichia coli and 86% homology to a gene product from Haemophilus influenzae . The E . coli gcpE gene was able to functionally complement the aarC1 allele in P . stuartii . The aarC1 allele was identified as a T to G transversion that resulted in a valine to glycine substitution at position 136 in the AarC protein . The aarC gene appears to be essential for cell viability as construction of a disrupted copy (aarC::lacZ) was possible only in cells that carried an episomal copy of aarC or gcpE.

J Bacteriol, 1997 Apr, 179(7), 2238 - 46
Sequence analysis and characterization of a 40-kilodalton Borrelia hermsii glycerophosphodiester phosphodiesterase homolog; Shang ES et al.; We report the purification, molecular cloning, and characterization of a 40-kDa glycerophosphodiester phosphodiesterase homolog from Borrelia hermsii . The 40-kDa protein was solubilized from whole organisms with 0.1% Triton X-100, phase partitioned into the Triton X-114 detergent phase, and purified by fast-performance liquid chromatography (FPLC) . The gene encoding the 40-kDa protein was cloned from a B . hermsii chromosomal DNA lambda EXlox expression library and identified by using affinity antibodies generated against the purified native protein . The deduced amino acid sequence included a 20-amino-acid signal peptide encoding a putative leader peptidase II cleavage site, indicating that the 40-kDa protein was a lipoprotein . Based on significant homology (31 to 52% identity) of the 40-kDa protein to glycerophosphodiester phosphodiesterases of Escherichia coli (GlpQ), Bacillus subtilis (GlpQ), and Haemophilus influenzae (Hpd; protein D), we have designated this B . hermsii 40-kDa lipoprotein a glycerophosphodiester phosphodiesterase (Gpd) homolog, the first B . hermsii lipoprotein to have a putative functional assignment . A nonlipidated form of the Gpd homolog was overproduced as a fusion protein in E . coli BL21(DE3)(pLysE) and was used to immunize rabbits to generate specific antiserum . Immunoblot analysis with anti-Gpd serum recognized recombinant H . influenzae protein D, and conversely, antiserum to H . influenzae protein D recognized recombinant B . hermsii Gpd (rGpd), indicating antigenic conservation between these proteins . Antiserum to rGpd also identified native Gpd as a constituent of purified outer membrane vesicles prepared from B . hermsii . Screening of other pathogenic spirochetes with anti-rGpd serum revealed the presence of antigenically related proteins in Borrelia burgdorferi, Treponema pallidum, and Leptospira kirschneri . Further sequence analysis both upstream and downstream of the Gpd homolog showed additional homologs of glycerol metabolism, including a glycerol-3-phosphate transporter (GlpT), a glycerol-3-phosphate dehydrogenase (GlpD), and a thioredoxin reductase (TrxB).

MMWR Recomm Rep, 1997 Mar 28, 46(RR-7), 1 - 25
Pertussis vaccination: use of acellular pertussis vaccines among infants and young children . Recommendations of the Advisory Committee on Immunization Practices (ACIP)
Purification and characterisation of a plasminogen-binding protein from Haemophilus influenzae . Sequence determination reveals identity with aspartase.
Department of Clinical Chemistry, Karolinska Hospital, Stockholm, SwedenPlasminogen binding proteins have been described both for Gram positive and Gram negative bacteria . In the present work we describe the purification and characterization of a plasminogen binding protein from Haemophilus influenzae (strain HI-23459) . Bacteria were sonicated in order to solubilize plasminogen-binding proteins . The supernatant was subjected to affinity chromatography on plasminogen kringle-4 fragment bound to Sepharose 4B and subsequently processed by ion-exchange chromatography on DEAE-Sepharose CL-6B . Characterization of the protein by SDS-PAGE displayed a single band with a molecular mass of about 55,000, both prior to and after reduction . The purified protein stimulates tPA (tissue plasminogen activator) catalysed plasminogen activation by a factor of approximately 300, mainly due to a decrease in K(m) . Antibodies were raised in rabbits and used in quantitative and qualitative analysis . However, using a FITC-conjugate we failed to demonstrate the presence of the purified protein on the surface of intact bacteria . The corresponding gene was isolated from a lambda EMBL3 phage library prepared from chromosomal DNA from the same H . influenzae strain, using an oligonucleotide probe based on the NH2-terminal amino acid sequence . An open reading frame corresponding to 472 amino acid was found . The amino acid sequence of the translated gene demonstrates 97% identity with the recently published sequence from aspartate ammonia lyase (aspartase) from H . influenzae . Enzymatic analysis of the purified protein revealed a high aspartase activity.

Arch Microbiol, 1997 Mar 7, 167(2/3), 151 - 9
The alternative sigma factor sigmaB in Staphylococcus aureus: regulation of the sigB operon in response to growth phase and heat shock
Kullik I I, Giachino P.
In the human pathogen Staphylococcus aureus, many proteins involved in the infection process are preferentially produced during the stationary growth phase . Using a DNA probe corresponding to the Bacillus subtilis gene encoding the stationary-phase sigma factor SigB (sigmaB), we identified a gene in S . aureus with similarity to B . subtilis sigB . The sigB region was mapped on the SmaI I fragment of the S . aureus chromosome and contains a total of six open reading frames (orf1-6) . The deduced amino acid sequences of orf2, orf3, orf4, and orf5 show 64, 67, 71, and 77% similarity to the B . subtilis proteins RsbU, RsbV, RsbW, and SigB, respectively, with SigB representing the sigma factor and the Rsb proteins representing regulators of sigma B . Furthermore, the relative position of the corresponding genes is conserved in B . subtilis, which strongly suggests that we identified the sigB operon of S . aureus, encoding an alternative sigma factor in this organism . The proposed gene products of the two remaining open reading frames show 48-62% similarity to the PemK, ChpAK, and ChpBK growth inhibitors of Escherichia coli (ORF1) and 61% similarity to the ribosomal protein S1 of Haemophilus influenzae (ORF6) . Northern blot analysis of the sigB region in S . aureus revealed that four different transcripts are expressed under different conditions of growth phase and stress . These results indicate a complex transcriptional regulation that differs between S . aureus and B . subtilis.

Int J Pediatr Otorhinolaryngol, 1997 Mar 6, 39(2), 119 - 31
A resolved pneumococcal infection protects against nontypeable Haemophilus influenzae: an evaluation of different routes of whole cell immunization in protection against experimental acute otitis media; Melhus A et al.; A conferred cross-protection between Haemophilus influenzae type b (Hib) and nontypeable H . influenzae (NTHi) was demonstrated in a previous study of experimental recurrent otitis media . To explore cross-protection further, and to compare oral administration of whole cells with two more conventional routes for vaccination against acute otitis media (AOM), a total number of 79 rats were immunized perorally, subcutaneously and intrabullarly with H . influenzae or pneumococci and thereafter challenged in the middle ear with NTHi or Hib 4 or 9 weeks later . Otomicroscopic changes, bacterial cultures, and serum IgG antibody levels were monitored . The study demonstrated that while peroral administration did not elicit any protection, a resolved pneumococcal AOM could reduce the susceptibility to reinfection with NTHi . In the latter case no cross-reacting antibodies were detected, but the protective rate was 50% or more, and it was comparable with that found after subcutaneous or intrabullar immunization with homologous NTHi or Hib strains . The results suggest that the protection of the rat middle ear mucosa may involve unspecific responses.

J Public Health Manag Pract, 1996 Fall, 2(4), 31 - 6
Overcoming barriers and reaping the benefits of surveillance for infectious diseases: the New Mexico perspective; Sewell CM; Surveillance for disease is one of the cornerstones of public health practice in the United States . Surveillance, particularly for infectious diseases, has allowed the detection of outbreaks and provided for the long-term monitoring of disease incidence . In New Mexico, acquired immunodeficiency syndrome (AIDS) surveillance is characterized as one of the most comprehensive surveillance systems for an infectious disease to be found anywhere . The success of this system is largely a result of state and federal resources and a good partnership with the AIDS/human immunodeficiency virus health care providers . Surveillance for Haemophilus influenzae type b (Hib) has demonstrated a remarkable decline in disease incidence in the state especially since the use of second generation Hib capsular polysaccharide conjugate vaccine . In contrast, surveillance for hepatitis A has demonstrated a significant public health problem that is largely not being addressed by current control measures.

Onderstepoort J Vet Res, 1997 Mar, 64(1), 57 - 63
Effects of growth conditions and incubation times on the expression of antigens of Haemophilus paragallinarum which are detected by monoclonal antibodies; Bragg RR et al.; Haemophilus paragallinarum causes infectious coryza in poultry, and a panel of monoclonal antibodies (Mabs) were established, which detect surface antigens of this bacterium . It was postulated that these Mabs could be used to detect antigenic differences between strains of H . paragallinarum used in infectious coryza (IC) vaccines, and isolates made from the field, from poultry vaccinated against IC . It has previously been reported that in South Africa there are three different Mab patterns that have been common to H . paragallinarum isolates for the past three decades . The effects of different growth conditions such as duration of incubation, inoculum size, levels of NAD or NaCl in the medium, and the pH of the medium on these Mab patterns were investigated . It was found that many different factors appear to influence the expression of the antigens detected by the panel of Mabs . It was found that at different stages during the growth cycles, the isolates could be classified into different Mab groups . It was also found that alteration of the inoculum size resulted in Mab-pattern switches . Addition of extra NaCl to the medium, in order to slow the growth rate, was found to result in Mab-pattern switches . pH was found to have significant effects on the levels of expression of the antigens detected by the Mabs, although these changes did not result in Mab-pattern switches . The effects of pH were also found to be highly strain dependent . The use of NAD, rather than sterile chicken serum, in the medium did not significantly alter the levels of expression of these antigens . Alterations of the growth conditions greatly affected the levels of expression of the antigens detected by the Mabs, and were highly strain dependent . It was not possible to predict the effects of a particular growth condition on a particular strain or isolate of H . paragallinarum.

Arch Pediatr, 1997 Mar, 4(3), 213 - 8
{Bacteriological study in acute otitis media}; Le Bideau M et al.; AIM: A prospective study on bacteriological epidemiology in acute otitis media was conducted in a pediatric hospital emergency service from January 1993 to October 1995 . PATIENTS: One hundred and fifty-eight children, aged 6 months to 6 years, with an acute otitis media were included . Culturing and cleansing of the ear canal and tympanocentesis for aspiration and culture of the secretions were performed in 118 children (46 of whom had received antibiotics before for 48 hours) . MAIN RESULTS: Middle ear aspirates were sterile in 35% of the children who had not received antibiotics and in 64% of those already treated . Bacteria in middle ear were predominantly Haemophilus influenzae and Streptococcus pneumoniae . Fifty-nine percent of S pneumoniae strains were penicillin-resistant; however, they were responsible for clinical failure in only 8% of cases . No Staphylococcus strains, commensal of the ear canal, could be considered as pathogenic for the middle ear . CONCLUSION: The preciseness with which secretions of middle ear are aspirated reduces the risk of contamination and comparison of ear canal and middle ear cultures allows to identify them . The high ratio of sterile middle ear aspirates after antibiotic treatment raises the question if other factors are responsible for persistent symptoms . The existence of penicillin-resistant S pneumoniae must be known to adjust treatment.

Ann Trop Paediatr, 1997 Mar, 17(1), 5 - 8
The increasing burden of disease in Bangladeshi children due to Haemophilus influenzae type b meningitis; Saha SK et al.; A laboratory-based study of diagnosed bacterial meningitis in the national paediatric hospital identified 852 cases of meningitis in the 8-year period 1987-1994 . There were 587 culture-positive cases, of which Haemophilus influenzae (47%) and Streptococcus pneumoniae (32%) accounted for 80% . H . influenzae showed a remarkable increase of 700% during the study period . Most of the H . influenzae cases (90%) occurred in the 1st 2 years of life . Analysis of culture-negative specimens by antigen detection in the last 2 years also revealed the predominance of H . influenzae (71.4%) and S . pneumoniae (22.4%) . Typing of H . influenzae isolates during this time showed that 98% of the strains were type b . This retrospective hospital-based study indicated a pronounced increase in the incidence of H . influenzae meningitis and strongly supports the need for large-scale Hib vaccination for young children . However, such nationwide intervention will probably need to be based on a prospective on a prospective and population-based surveillance of H . influenzae infections.

Electrophoresis, 1997 Mar-Apr, 18(3-4), 432 - 42
Probing protein function using a combination of gene knockout and proteome analysis by mass spectrometry; Dainese P et al.; Recently the determination of the genome sequences of three procaryotes (Haemophilus influenzae, Methanococcus jannaschii and Mycoplasma genitalium) as well as the first eucaryotic genome (Saccharomyces cerevisiae) were completed . Between 40-60% of the genes were found to code for proteins to which no function could be assigned . We describe an approach which combines proteome analysis (mapping of expressed proteins isolated by two-dimensional polyacrylamide gel electrophoresis to the genome) with genetic manipulations to study the complex pattern of protein regulation occurring in Escherichia coli in response to sulfate starvation . We have previously described the upregulation of eight spots on two-dimensional (2-D) gels in response to sulfate starvation and the assignment of six of these to entries in the E . coli genome sequence (Quadroni et al., Eur . J . Biochem . 1996, 239, 773-781) . Here we describe the identification of the remaining two proteins which are encoded in a sulfate-controlled operon in the 21.5' region of the E . coli genome . Upregulated protein spots were cut from multiple 2-D gels collected and run on a modified funnel gel to concentrate the proteins and remove the sodium dodecyl sulfate before digestion . The peptide masses obtained from the digests were used to search the SwissProt database or a six-frame translation of the EMBL DNA database using a peptide mass fingerprinting algorithm . A digest can be reanalyzed after deuterium exchange to obtain a second, orthogonal data set to increase the confidence level of protein identification . The digests of the remaining unidentified proteins were used for peptide fragment generation using either post-source decay in a matrix-assisted laser desorption ionization (MALDI) time-of-flight mass spectrometer or collision-induced dissociation (CID) coupled mass spectrometry (MS/MS) with triple stage quadrupole or ion trap mass spectrometers . The spectra were used as peptide fragment fingerprints to search the SwissProt and EMBL databases.

Diagn Microbiol Infect Dis, 1997 Mar, 27(3), 75 - 83
A 1994-95 survey of Haemophilus influenzae susceptibility to ten orally administered agents . A 187 clinical laboratory center sample in the United States; Jones RN et al.; During August, 1994 to April, 1995, a total of 2278 clinical isolates of Haemophilus influenzae were obtained from 187 clinical laboratories in the United States (U.S.) . The vast majority of these isolates (75%) were from respiratory sites, and the remaining organisms were from blood, ear, eye, and spinal fluid sources . The overall rate of beta-lactamase production and ampicillin resistance was 36% . The antimicrobial susceptibility of isolates was determined by reference broth microdilution testing against ten orally administered agents . MIC values were compared according to 12 geographical regions, inpatient or outpatient status, gender, and eight age groupings . Modest and occasionally significant differences were observed: 1) greater numbers of beta-lactamase-producing strains among outpatients, in males, in the mid-Atlantic region, and in children < or = 12 years of age; 2) lower prevalence of beta-lactamase-producing isolates in the Southeast and Pacific regions; 3) cefaclor, cefprozil, and loracarbef activity was lowest among the younger children (< or = six years); and 4) macrolide in vitro efficacy was lowest in patients > 50 years of age and in three eastern regions . Overall, more than 99% of the strains were susceptible to amoxicillin/clavulanic acid, cefixime, and cefpodoxime (e.g., widest potential clinical use) . Susceptibilities using National Committee for Clinical Laboratory Standards (NCCLS) breakpoint criteria for the other agents were: 96.6% to cefuroxime, 86.5% to loracarbef, 84.0% to clarithromycin, 81.8% to cefaclor, and 80.7% to cefprozil . Non-beta-lactamase mechanisms of resistance to ampicillin were rare (0.2%) or episodic and were attributed to altered penicillin-binding proteins . Although there is an increased prevalence of beta-lactamase production among H . influenzae isolates compared to prior years, four beta-lactams remain highly active (> 95% susceptibility) against contemporary strains of H . influenzae . Other monitored compounds seem to have declined in spectrum and surveillance trials for resistance among H . influenzae isolates should continue in an effort to identify trends in the U.S.

Br J Rheumatol, 1997 Mar, 36(3), 370 - 3
Bacterial joint infections in England and Wales: analysis of bacterial isolates over a four year period; Ryan MJ et al.; Data from 1158 cases of septic arthritis reported to the Public Health Laboratory Service (PHLS) Communicable Disease Control Centre (CDSC) from England and Wales over a 4 yr period (January 1990 December 1993) are presented . Reports where a bacterial organism was isolated from synovial fluid, or where an organism was isolated from blood cultures where a diagnosis of septic arthritis was reported, were examined . Reports of infection were more common in children (12.7% of infections were in the under 10 age group) and the elderly (54.7% aged 60 or over), and were higher in males in all age groups except in the elderly . The most common causative organisms remain staphylococcal and streptococcal species, comprising 40.6% (470) and 28% (324) of cases, respectively . The most common streptococci seen were Streptococcus pneumoniae and Lancefield group A beta-haemolytic Streptococcus organisms, 60.8% (197/324), although group B, C and G organisms accounted for 33.6% of streptococcal isolates (109/324) . Haemophilus influenzae septic arthritis is not exclusive to children as 23.2% (16-69) of cases occurred over the age of 15 . A total of 48% (635) of isolates were identified from both synovial fluid and blood cultures, 32.6% (378) from joint fluid alone and 12.5% (146) from blood cultures . Although this study excludes cases of septic arthritis where no organism was isolated, it presents important bacteriological information from a large number of isolates from England and Wales over a 4 yr period . Risk factors identified include a joint prosthesis, joint disease/connective tissue disorder . immunosuppression and diabetes.

Am J Rhinol, 1997 Mar-Apr, 11(2), 139 - 43
Correlation of middle meatal and maxillary sinus cultures in acute maxillary sinusitis; Vaidya AM et al.; Cultures obtained from the middle meatus have been used frequently in the past to direct therapy in patients with acute maxillary sinusitis . However, no convincing data have been published to indicate that middle meatal cultures accurately represent the bacterial flora within the maxillary sinus . The hypothesis of this experiment is that bacteria obtained by directed middle meatal cultures qualitatively and quantitatively correlate with cultures taken by maxillary sinus puncture . Acute sinusitis was induced by injecting 10(8) colony-forming units of bacteria directly into the maxillary sinuses of rabbits in which the ostia were occluded with cotton packs . Eight animals were injected with Staphylococcus aureus, eight with Haemophilus influenzae, and eight with Streptococcus pneumoniae . The packs were removed after 3 days, and specimens were obtained from the middle meatus in the region of the maxillary sinus ostium, and from the maxillary sinus, 1 day later . The contralateral maxillary sinuses of six of the animals were injected with normal saline and served as controls . There was a 100% correlation rate between cultures of specimens obtained from the maxillary sinus and from the middle meatus in all 24 animals . In addition, the quantitative counts from the middle meatus and the maxillary sinus correlated . Control animals showed no bacterial growth from either the middle meatus or the maxillary sinus . These results show that, in an animal model of acute sinusitis, cultures of specimens from the middle meatus reflect the contents of the maxillary sinus.

J Med Microbiol, 1997 Mar, 46(3), 204 - 7
Pyrolysis mass spectrometry in epidemiological and population genetic studies of Haemophilus influenzae; Leaves NI et al.; Haemophilus influenzae serotype b (Hib) vaccines have reduced the amount of invasive Hib disease in immunised infants . However, Hib disease remains in unvaccinated infants and adults and non-capsulate H . influenzae (NCHi) still causes infections, including outbreaks of respiratory disease . Characterisation of strains and the bacterial population as a whole is therefore necessary to detect outbreaks of infection with NCHi or changes in the population, for example, to vaccine-resistant clones of Hib . The rapid, simple and objective technique of pyrolysis mass spectrometry (PMS) was investigated as an alternative to current complex, subjective methods . PMS was compared with ribotyping and multilocus enzyme electrophoresis (MLEE) for population genetic analyses of Hib and with ribotyping and protein profiling for epidemiological analyses of NCHi . PMS clustered all the isolates of Hib together whereas MLEE and ribotyping distinguished certain clones - this is probably because the three methods examine different (and unrelated) characteristics of the organisms . The PMS results were essentially similar to those from ribotyping and protein profiling for the epidemiological analyses of outbreaks of NCHi disease . Therefore, PMS is probably unsuitable for comparisons of Hib populations but it is a useful addition to the arsenal of techniques for the characterisation of NCHi.

Otolaryngol Head Neck Surg, 1997 Mar, 116(3), 363 - 71
Immunoglobulin G subclass response to major outer membrane proteins of nontypable Haemophilus influenzae in children with acute otitis media; Bernstein JM et al.; Children with acute otitis media as the result of nontypable Haemophilus influenzae often develop serum bactericidal and/or opsonic IgG antibodies to this organism during convalescence . Outer membrane proteins appear to be the principal targets for such antibodies . In this study we characterized the IgG subclass responses to major outer membrane proteins of nontypable H . influenzae in otitis-prone children in whom this organism had colonized . Three of the major outer membrane proteins (P2, P5, and P6) were isolated from the homologous nontypable H . influenzae strain recovered from the middle ear at the time of acute infection . Sera were obtained during the acute phase and at 1 and 6 months thereafter . The outer membrane proteins, which were isolated by preparative sodium dodecylsulfate-polyacrylamide gel electrophoresis, were used as test antigens in a quantitative IgG subclass enzyme immunoassay . The results of this analysis indicate that the temporal characteristics and distribution of IgG subclass antibodies were found to differ for each of the outer membrane proteins . Moreover, substantial variation between patients was observed with respect to both temporal characteristics and subclass distribution of the IgG response to the three outer membrane proteins . Significantly, sera from two of three otitis-prone subjects contained detectable levels of IgG antibody to the conserved P6 outer membrane protein at the time of acute infection, with serum from one subject also containing detectable levels of IgG3 antibody to this same protein . Nevertheless, the organism persisted in the middle ears of these patients . The results of this study indicate that otitis-prone children manifest a highly variable IgG subclass response to both conserved (P6) and variable (P2) outer membrane proteins of nontypable H . influenzae . Further study is required to ascertain whether these IgG subclass antibodies are biologically efficacious and whether otitis-prone children possess the immunologic maturity to respond to nontypable H . influenzae outer membrane protein-based vaccines in a predictable manner.

Otolaryngol Head Neck Surg, 1997 Mar, 116(3), 301 - 7
Adenoid bacteriology and sinonasal symptoms in children; Lee D et al.; OBJECTIVE: To study the correlation between sinonasal symptoms in children and the prevalence of bacterial pathogens in the adenoid core . DESIGN: Prospective cross-sectional survey . SETTING: Tertiary referral center . PATIENTS: Consecutive sample of 84 children, aged 2 to 12 years, scheduled for adenoidectomy between July 1995 and November 1995 . A blocked recruitment scheme was used to ensure a balanced distribution of sinonasal symptoms in the study sample . METHODS: A caregiver-completed quality-of-life survey was used to measure the frequency and severity of baseline sinonasal, ear, obstructive, and behavioral symptoms . Standard microbiologic techniques were used for qualitative and quantitative adenoid bacteriology . OUTCOME MEASURES: Pathogenic and nonpathogenic bacteria isolated, percentage of specimens with no growth, colony-forming units of pathogens per gram of adenoid tissue, dominant bacterial organism, and number of pathogenic species per adenoid with concentration greater than 10(5) colony-forming units . RESULTS: One or more bacterial pathogens were recovered from core samples of all adenoids, with a concentration greater than 10(5) colony-forming units in 31 specimens (26%) . Haemophilus influenzae, group A beta-hemolytic streptococcus, and Staphylococcus aureus were encountered most often . Multivariate analysis revealed a significant correlation of sinonasal infection symptom scores with colony-forming units of adenoid core pathogens (R2 = 0.48, p < 0.0001), adjusted for the confounding effects of nasal obstructive symptoms and adenoid size (specimen weight) . CONCLUSIONS: Sinonasal infectious symptoms explain 48% of the variability in quantitative bacteriology of the adenoid core, independent of adenoid size . Although longitudinal studies are required, our results support a potential role for adenoidectomy in the management of refractory pediatric sinusitis.

Arzneimittelforschung, 1997 Mar, 47(3), 299 - 302
Cefodizime given in single or divided doses for the treatment of lower respiratory tract infection; Palombini BC et al.; The safety and efficacy of two dose regimens of cefodizime (CAS 69739-16-8, HR 221) in hospitalized patients with lower respiratory tract infections were assessed in two consecutive studies . Sputum bacteriology, chest X-ray and a safety laboratory check were performed at baseline and after therapy . In order to compensate for the lack of a double-blind design the evaluation was conducted as a clinical intention-to-treat analysis . 32 patients (16 males, 16 females, mean age: 64 years) were admitted to study A and 42 subjects (30 males, 12 females, mean age: 66 years) to study B . The dosage regimens of cefodizime were 1 g b.i.d . (median 7 days) in study A and 2 g once daily (median 6 days) in study B . Parenchymal involvement was confirmed by chest X-ray in 56% of the patients in study A and 64% in study B, the remainder patients had acute exacerbations of chronic bronchitis with reasonable evidence of bacterial infection . The most frequent pathogens were Streptococcus pneumoniae and Haemophilus spp . The clinical cure rate was 97% in study A and 88% in study B . Eradication rates were 100% and 94%, respectively . No superinfection occurred . No adverse reactions were observed . In conclusion, a single daily injection of cefodizime 2 g achieved similar clinical and bacteriological cure rates to the standard dose regimen of 1 g b.i.d.

J Antimicrob Chemother, 1997 Mar, 39(3), 415 - 8
Pharmacodynamic properties of faropenem demonstrated by studies of time-kill kinetics and postantibiotic effect; Boswell FJ et al.; The pharmacodynamic properties of faropenem, a new oral penem antibiotic, were investigated by studying time-kill kinetics and postantibiotic effect . Time-kill kinetics were employed against strains of Bacteroides fragilis, Escherichia coli, Staphylococcus aureus, Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pyogenes . The postantibiotic effects of faropenem were studied using strains of E . coli, S . aureus, H . influenzae and Streptococcus pneumoniae . The time-kill kinetic data demonstrated that faropenem has bactericidal activity . Faropenem exhibited a significant postantibiotic effect against all strains except H . influenzae.

J Antimicrob Chemother, 1997 Mar, 39(3), 411 - 4
Multicentre in-vitro evaluation of the susceptibility of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis to ciprofloxacin, clarithromycin, co-amoxiclav and sparfloxacin; Hoogkamp-Korstanje JA et al.; Seven laboratories, including a reference laboratory, tested the susceptibility of Moraxella catarrhalis, Streptococcus pneumoniae and Haemophilus influenzae strains to ciprofloxacin, clarithromycin, co-amoxiclav and sparfloxacin with the Etest . A total of 976 strains were collected . The results with ciprofloxacin and sparfloxacin were consistent for all laboratories, while those with clarithromycin and co-amoxiclav were not . The agreement between Etest MICs and broth microdilution was: ciprofloxacin and sparfloxacin, >95%; clarithromycin for all species, 71-85%; co-amoxiclav for H . influenzae, 31% . MIC90 values (broth dilution, mg/L) for M . catarrhalis, S . pneumoniae and H . influenzae were: sparfloxacin, 0.06, 0.5, 0.03; ciprofloxacin, 0.12, 2.0, 0.03; co-amoxiclav, 0.25, 0.25, 0.25; clarithromycin 0.25, 0.25 and 16.

J Antimicrob Chemother, 1997 Mar, 39(3), 303 - 7
A molecular analysis of Greek and UK Haemophilus influenzae conjugative resistance plasmids; Dimopoulou ID et al.; Antibiotic resistance in Haemophilus influenzae has been associated with the presence of large, chromosomally integrated, conjugative plasmids . The plasmids of 10 beta-lactamase-positive, ampicillin-resistant strains, two from the UK and eight from Greece, were investigated . Plasmids were detected and isolated after transfer to a rec-deficient recipient . Purified whole plasmid was used as probe . In addition a 12 kb PstI fragment containing the putative point of recircularization in one plasmid, p1056, was cloned and used as a probe . All plasmids shared a high degree of sequence homology suggesting that plasmids of diverse geographical origin are highly related . All plasmids also shared sequence homology with the 12 kb PstI fragment containing the point of recircularization, suggesting that the sequences involved in excision and recircularization are conserved.

Pharmacotherapy, 1997 Mar-Apr, 17(2), 235 - 41
Pharmacodynamic activity of five oral cephalosporins against Haemophilus influenzae; Stein GE et al.; STUDY OBJECTIVES: To determine the time above minimum inhibitory concentration (T > MIC) and serum bactericidal activity of five oral cephalosporins against two strains of Haemophilus influenzae . DESIGN: Randomized, crossover study . SETTING: University-associated research center . SUBJECTS: Ten healthy volunteers . INTERVENTIONS: Each subject received a single dose of cefpodoxime 200 mg, cefuroxime 500 mg, cefaclor 500 mg, cefprozil 500 mg, or loracarbef 400 mg each week for 5 weeks . Blood for serum levels was obtained at time zero and 1, 2, 3, 4, 6, 8, and 12 hours after each dose . MEASUREMENTS AND MAIN RESULTS: Cefpodoxime produced serum concentrations above the MIC for more than 90% of the time for both beta-lactamase-negative and -positive strains of H . influenzae . Moreover, it had serum bactericidal activity for 12 hours against both isolates . Cefuroxime was the second most active cephalosporin, with serum concentrations above the MIC of both isolates for 60% of the time . Cefuroxime provided serum bactericidal activity for 12 hours against the beta-lactamase-negative strain and 6 hours against the beta-lactamase-positive strain of H . influenzae . Even though the T > MIC was less than 50% of the study period for the other cephalosporins, all but cefaclor provided serum bactericidal activity for 12 hours against the beta-lactamase-negative isolate . Cefaclor provided measurable serum bactericidal activity for only 3 hours . The duration of serum bactericidal activity of cefprozil, loracarbef, and cefaclor against the beta-lactamase-positive isolate was 4, 2, and 0 hours, respectively . CONCLUSION: Cefpodoxime was the most active cephalosporin studied based on T > MIC and serum bactericidal activity against isolates of H . influenzae.

Microbiology, 1997 Mar, 143 ( Pt 3), 899 - 907
Molecular genetic analysis of the region containing the essential Pseudomonas aeruginosa asd gene encoding aspartate-beta-semialdehyde dehydrogenase; Hoang TT et al.; asd mutants of Gram-negative and some Gram-positive bacteria have an obligate requirement for diaminopimelic acid (DAP), an essential constituent of the cell wall of these organisms . In environments deprived of DAP, for example mammalian tissues, they will undergo lysis . This was previously exploited to develop vaccine strains of Salmonella typhimurium and cloning vectors containing asd as an in vivo selectable marker . As a first step for development of such systems for Pseudomonas aeruginosa, the asd gene from wild-type strain PAO1 was cloned by a combined approach of PCR amplification from chromosomal DNA, construction of mini-libraries and by complementation of an Escherichia coli delta asd mutant . The nucleotide sequence of a 2433 bp Smal-Nsil fragment was determined . This fragment contained the C-terminal 47 nucleotides of leuB, encoding 3-isopropylmalate dehydrogenase; asd, encoding aspartate-beta-semialdehyde dehydrogenase (Asd); and orfA, whose product showed similarity to the Asd proteins from Vibrio spp . By subcloning, asd was localized to a 1.24 kb DNA fragment which in an E . coli T7 expression system strongly expressed a 40,000 Da protein . The amino acid sequence was deduced from the DNA sequence . A comparison of the Asd proteins from P . aeruginosa, E . coli and Haemophilus influenzae revealed greater than 63% identity, demonstrating the conserved nature of Asd in Gram-negative bacteria, and defined the active-site-containing consensus sequence GGNCTVXMLMXXXLGLF as a possible signature motif . Chromosomal delta asd mutants were isolated . They were auxotrophic for DAP, lysine, methionine and threonine, and lysed in the absence of DAP . Genetic analyses indicated that orfA probably is naturally frame-shifted and does not contribute to the Asd phenotype . By PFGE, the asd gene was mapped to between coordinates 1.89 and 2.15 Mbp, or 37-40 min, on the 5.9 Mbp P . aeruginosa chromosome.

Pediatr Infect Dis J, 1997 Mar, 16(3 Suppl), S60 - 4
Empiric antibiotic selection criteria for respiratory infections in pediatric practice; Pichichero ME; BACKGROUND: Respiratory infections in children may occur as a consequence of resistant bacterial pathogens . Streptococcus pneumoniae organisms resistant to penicillin, trimethoprim/sulfamethoxazole and macrolides are increasingly prevalent . Amoxicillin- and macrolide-resistant Haemophilus influenzae and Moraxella (Branhamella) catarrhalis are also more commonly seen . Traditional agents such as amoxicillin and trimethoprim/sulfamethoxazole remain acceptable choices for most children with respiratory infections because currently most patients are not infected by resistant pathogens and there is a high spontaneous cure rate associated with these infections . OBJECTIVE: To analyze the criteria for the selection of extended spectrum antimicrobials as empiric therapy for respiratory infections . DISCUSSION: When an extended spectrum antimicrobial is appropriate for empiric therapy, selection should be based on: (1) efficacy; (2) adverse event profile; and (3) compliance-enhancing features (dosing with meals, once or twice daily administration, good palatability in suspension, shortened course of therapy and affordability) . A new agent, ceftibuten, has recently joined other extended spectrum cephalosporins and newer macrolides (clarithromycin and azithromycin) as a choice to be considered for empiric therapy for respiratory infections . These antimicrobials are differentiated from each other and traditional agents by differences in activity in vitro against penicillin-resistant pneumococci, relative beta-lactamase stability against Gram-negative bacteria and pharmacodynamic properties . When resistant organisms are isolated or suspected in community-acquired respiratory infections, cautious use of newer antibiotics may have to be considered.

Drugs, 1997 Mar, 53(3), 349 - 57
Choosing the right macrolide antibiotic . A guide to selection; Charles L et al.; Macrolide antibiotics have proven to be valuable alternatives to penicillins and cephalosporins for the treatment of a number of infections . Currently, a number of macrolides are available . When choosing a particular macrolide, the types of organisms causing the infection, the tolerability of the drug, convenience of dosing and possible drug interactions all must be taken into account . Erythromycin, azithromycin and clarithromycin are equally effective against most gram-positive organisms . However, clarithromycin and azithromycin have much better activity against Haemophilus influenza and Moraxella catarrhalis . Thus, these 2 drugs are better choices for the treatment of community-acquired pneumonia . However, the low serum concentrations of azithromycin may be a problem in patients with bacteraemia associated with with community-acquired pneumonia . Clarithromycin appears to be effective for the treatment and prophylaxis of Mycobacterium avium complex (MAC) in patients with AIDS, while azithromycin appears to be effective for prophylaxis . Treatment of MAC with azithromycin is currently undergoing study . Although clarithromycin is the macrolide of choice for the treatment of Helicobacter pylori, azithromycin is the preferred macrolide for the treatment of Chlamydia trachomatis infections . The major factor limiting the use of azithromycin and clarithromycin has been their cost . However, these drugs may be cost effective if compliance is improved due to better tolerability and more convenient dosing regimens.

J Bacteriol, 1997 Mar, 179(6), 1872 - 9
Cloning and characterization of bacteriophage-like DNA from Haemophilus somnus homologous to phages P2 and HP1; Pontarollo RA et al.; In an attempt to identify and characterize components of a heme uptake system of Haemophilus somnus, an Escherichia coli cosmid library of H . somnus genomic DNA was screened for the ability to bind hemin (Hmb+) . The Hmb+ phenotype was associated with a 7,814-bp HindIII fragment of H . somnus DNA that was subcloned and sequenced . Thirteen open reading frames (orfs) were identified, all transcribed in one direction, and transposon mutagenesis identified orf7 as the gene associated with the Hmb+ phenotype . Orf7 (178 amino acids) has extensive homology with the lysozymes of bacteriophages P-A2, P21, P22, PZA, phi-29, phi-vML3, T4, or HP1 . The orf7 gene complemented the lytic function of the K gene of phage P2 and the R gene of phage lambda . A lysozyme assay using supernatants from whole-cell lysates of E . coli cultures harboring plasmid pRAP501 or pGCH2 (both of which express the orf7 gene product) exhibited significant levels of lysozyme activity . The orf6 gene upstream of orf7 has the dual start motif common to the holins encoded by lambdoid S genes, and the orf6 gene product has significant homology to the holins of phages HP1 and P21 . When expressed from a tac promoter, the orf6 gene product caused immediate cell death without lysis, while cultures expressing the orf7 gene product grew at normal rates but lysed immediately after the addition of chloroform . Based on this data, we concluded that the Hmb+ phenotype was an artifact resulting from the expression of cloned lysis genes which were detrimental to the E . coli host . The DNA flanking the cloned lysis genes contains orfs that are similar to structural and DNA packaging genes of phage P2 . Polyclonal antiserum against Orf2, which is homologous to the major capsid precursor protein (gpN) of phage P2, detected a 40,000-M(r) protein expressed from pRAP401 but did not detect Orf2 in H . somnus, lysates . The phage-like DNA was detected in the serum-susceptible preputial strains HS-124P and HS-127P but was absent from the serum-resistant preputial strains HS-20P and HS-22P . Elucidation of a potential role for this cryptic prophage in the H . somnus life cycle requires more study.

Clin Diagn Lab Immunol, 1997 Mar, 4(2), 168 - 72
Characterization of specific immunoglobulin G (IgG) and its subclasses (IgG1 and IgG2) against the 23-valent pneumococcal vaccine in a healthy adult population: proposal for response criteria; Rodrigo MJ et al.; The aim of the study was to standardize an enzyme-linked immunosorbent assay (ELISA) method for the quantification of immunoglobulin G (IgG) and its subclasses (IgG1 and IgG2) against the 23-valent pneumococcal vaccine and to establish the criteria for a normal response to the vaccine . Forty healthy individuals (20 women and 20 men; mean age, 29 years) were studied . All were vaccinated with the 23-valent pneumococcal vaccine; blood samples were drawn just prior to and 3 weeks after immunization . Quantification of specific IgG and its subclasses was performed by an ELISA with the vaccine as the antigen . The linearity of the ELISA method was demonstrated by the similar slopes of the linear regression lines generated from the titration of sera with different antibody concentrations . The specificity of the antibodies against the vaccine was demonstrated by (i) an absorption test with pneumococcal vaccine, (ii) a cross-reactivity experiment with Haemophilus influenzae type b polysaccharide, and (iii) affinity chromatography with protein A-Sepharose . Response to the vaccine was defined by using the lower level of the 90% probability interval (one-tailed) for postimmunization-specific IgG, IgG1, and IgG2 . By using this cutoff, responders were considered to be those with an absolute increase in antibody titers higher than 395 arbitrary units/ml for IgG, 0.350 A450 units for IgG1, and 0.314 A450 units for IgG2 . Overall, 20 (50%) subjects had IgG, IgG1, and IgG2 responses, 9 (22.5%) had IgG and IgG2 responses, 4 (10%) had IgG1 responses, 3 (7.5%) had IgG and IgG1 responses, and 4 (10%) were nonresponders . Ninety percent of our population responded to the 23-valent pneumococcal vaccine . Up to 10% of healthy individuals may respond to an IgG subclass without significant increases in total IgG titers . The ELISA method that is described may be useful for evaluating the specific antibody response against polysaccharides.

J Bacteriol, 1997 Mar, 179(5), 1764 - 73
The major outer membrane protein of Haemophilus ducreyi consists of two OmpA homologs; Klesney-Tait J et al.; The major outer membrane protein (MOMP) of Haemophilus ducreyi is an OmpA homolog that migrates on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels as three species with apparent molecular weights ranging from 37,000 to 43,000 . Monoclonal antibodies directed against this macromolecule were used to identify recombinant clones containing fragments of the gene encoding this protein . Nucleotide sequence analysis of these fragments confirmed that the MOMP encoded by the intact gene (momp) was a member of the OmpA family of outer membrane proteins . Construction of an isogenic H . ducreyi mutant unable to express the MOMP led to the discovery of a second outer membrane protein which migrated at the same rate on SDS-PAGE gels as the MOMP . N-terminal amino acid sequence analysis of this second protein revealed that its N terminus was nearly identical to that of the MOMP and also had homology with members of the OmpA family . Nucleotide sequence analysis of the region downstream from the momp gene revealed the presence of a partial open reading frame encoding a predicted OmpA-like protein . A modification of anchored PCR technology was used to obtain the nucleotide sequence of this downstream gene which was shown to encode a second OmpA homolog (OmpA2) . The N-terminal amino acid sequence of OmpA2 was identical to that of the OmpA-like protein detected in the momp mutant . The H . ducreyi MOMP and OmpA2 proteins, which comigrated on SDS-PAGE gels and which were encoded by the tandem arranged momp and ompA2 genes, were 72% identical.

J Bacteriol, 1997 Mar, 179(5), 1684 - 9
Cloning and characterization of the haemocin immunity gene of Haemophilus influenzae; Murley YM et al.; The bacteriocin haemocin is produced by most type b strains of Haemophilus influenzae, including strains of diverse genetic lineage, and is toxic to virtually all nontypeable H . influenzae strains . An H . influenzae transformant bearing a plasmid with a 1.5-kbp chromosomal fragment capable of conferring haemocin immunity on a haemocin-susceptible H . influenzae mutant was selected by using partially purified haemocin . Deletional and site-directed mutagenesis localized the haemocin immunity gene to the 3' open reading frame (ORF) within this chromosomal fragment . Subcloning of this ORF demonstrated that it was sufficient to confer haemocin immunity on wild-type haemocin-susceptible H . influenzae strains as well as haemocin-susceptible strains of Escherichia coli . This ORF, designated hmcl, encodes a 105-amino-acid protein with an estimated molecular mass of 12.6 kDa . Primer extension analysis revealed a putative transcriptional start site 34 bp upstream of the start codon, and the presence of a promoter immediately upstream of hmcI was confirmed by cloning the gene into a promoterless chloramphenicol acetyltransferase vector . To characterize the hmcI gene product, a His-HmcI fusion protein was constructed.

J Infect Dis, 1997 Mar, 175(3), 583 - 9
Comparison of clinical diagnosis and standard laboratory and molecular methods for the diagnosis of genital ulcer disease in Lesotho: association with human immunodeficiency virus infection; Morse SA et al.; A multiplex polymerase chain reaction (M-PCR) assay for Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus (HSV) was compared with clinical and standard laboratory methods for the diagnosis of genital ulcer disease (GUD) in 105 patients; 36% were human immunodeficiency virus (HIV)-seropositive . Chancroid (80%), syphilis (8%), and genital herpes (8%) were the most frequent diagnoses . H . ducreyi and HSV were isolated from ulcers of 43% and 18% of patients, respectively; in 35%, all cultures were negative and the laboratory diagnosis indeterminate . M-PCR detected H . ducreyi, T . pallidum, and HSV in 56%, 23%, and 26% of patients, respectively; (no definitive diagnosis, 6%) . The proportion of patients with more than one agent was 4% by culture and 17% by M-PCR (P = .002) . Resolved sensitivities of M-PCR for H . ducreyi and HSV cultures were 95% and 93%, respectively . The sensitivities of H . ducreyi and HSV cultures were 75% and 60%, respectively . HSV, detected in 47% of specimens from HIV-infected versus 16% from HIV-uninfected patients (P < .001), may be emerging as a more frequent cause of GUD.

Infect Immun, 1997 Mar, 65(3), 943 - 50
Decoration of lipopolysaccharide with phosphorylcholine: a phase-variable characteristic of Haemophilus influenzae; Weiser JN et al.; Choline, although not a nutritional requirement for Haemophilus influenzae, is taken up from the growth medium and incorporated into its lipopolysaccharide (LPS) . Incorporated choline is in the form of phosphorylcholine (ChoP) based on the reactivity with the monoclonal antibody with specificity for this structure, TEPC-15 . Incorporation of {3H}choline from the growth medium and expression of the TEPC-15 epitope undergo high-frequency phase variation, characteristic of other LPS structures in this species . The expression and phase variation of ChoP require a previously identified locus involved in LPS biosynthesis, lic1 . The first gene in lic1, licA, contains a translational switch based on variation in the number of intragenic tandem repeats of the sequence 5'-CAAT-3' . The full-length LicA polypeptide resembles choline kinases of eucaryotes, suggesting that the pathway for choline incorporation into the H . influenzae glycolipid has similarities to the pathway for choline incorporation in eucaryotic lipid synthesis . The display of ChoP, a host-like structure, renders the organism more rather than less susceptible to the bactericidal activity of human serum . The increased serum sensitivity of variants with ChoP correlates with higher serum immunoglobulin G titers to LPS containing this structure . ChoP appears to be a cell surface feature common to a number of pathogens of the human respiratory tract, including Streptococcus pneumoniae and mycoplasmas . In the case of H . influenzae, its primary contribution to pathogenesis does not appear to be antigenic variation to evade host humoral clearance mechanisms.

Infect Immun, 1997 Mar, 65(3), 877 - 81
Intrathecal production of interleukin-12 and gamma interferon in patients with bacterial meningitis; Kornelisse RF et al.; To assess the role of interleukin-12 (IL-12) and gamma interferon (IFN-gamma) in children with bacterial meningitis, bioactive IL-12 (p70) and the inactive subunit p40 and IFN-gamma were measured in serum and cerebrospinal fluid (CSF) from 35 children with bacterial meningitis and 10 control subjects . The production of IFN-gamma is induced by IL-12 with tumor necrosis factor alpha (TNF-alpha) as a costimulator and inhibited by IL-10 . CSF concentrations of IL-12 p40 as well as those of IFN-gamma were markedly elevated, whereas IL-12 p70 was hardly detectable . Detectable CSF levels of IFN-gamma correlated positively with IL-12 p40 (r = 0.40, P = 0.02) and TNF-alpha (r = 0.46, P = 0.04) but not with IL-6, IL-8, or IL-10 . In contrast to CSF levels of TNF-alpha, IL-12, and IL-10, those of IFN-gamma were significantly higher in patients with pneumococcal meningitis than in children with meningitis caused by Haemophilus influenzae and Neisseria meningitidis, presumably because of a high CSF TNF-alpha/IL-10 ratio in the former . We suggest that IL-12- and TNF-alpha-induced IFN-gamma production may contribute to the natural immunity against microorganisms in the CSF compartment during the acute phase of bacterial meningitis.

Biochemistry, 1997 Feb 25, 36(8), 2091 - 103
Structure of the variable and conserved lipopolysaccharide oligosaccharide epitopes expressed by Haemophilus influenzae serotype b strain Eagan; Masoud H et al.; Lipopolysaccharide (LPS) is a major virulence determinant of Haemophilus influenzae . The organism is capable of expressing a heterogeneous population of LPS which exhibits extensive antigenic diversity among multiple oligosaccharide (OS) epitopes . Structural elucidation of variable and conserved OS epitopes of H . influenzae serotype b strain Eagan was determined by the application of high-field NMR techniques and MS-based methods on oligosaccharides obtained from LPS samples by a deacylation strategy . LPS extracted by the hot aqueous phenol method gave complex electrophoretic patterns consisting of at least six low-molecular mass bands . Electrospray ionization-mass spectrometry of O-deacylated LPS revealed a series of related structures differing in the number of hexose residues as well as subpopulations of glycoforms containing additional phosphoethanolamine (PEA) groups . It was demonstrated that the LPS contains a conserved PEA-substituted, heptose-containing trisaccharide inner core moiety attached via a KDO 4-phosphate unit to a lipid A component . Tandem MS experiments unambiguously established the presence of a KDO 4-pyrophosphoethanolamine unit in the subpopulation of LPS containing additional PEA groups . The occurrence of LPS containing this structural feature was found to be dependant on the isolation procedure used . Each heptose of the common inner core element L-alpha-D-Hepp(1-->2)-L-alpha-D-Hepp(1-->3)-L-alpha-D-Hep p(1-->5)-alpha-KDO is substituted by a hexose residue with further chain elongation from the central unit . The structures of the major glycoforms containing four (three Glcs and one Gal), five (three Glcs and two Gals), and six (three Glcs and three Gals) hexoses were determined in detail . The Hex6 glycoform contains the terminal structure, alpha-D-Galp(1-->4)-beta-D-Galp(1-->4)-beta-D-Glc, providing, for the first time, definitive structural evidence for the expression of the Pk-blood group antigen in H . influenzae LPS . Moreover, an analogue of the Hex4 glycoform was identified in which the third heptose residue carries phosphate at 0-4.

Arch Intern Med, 1997 Feb 24, 157(4), 425 - 30
Acute bacterial meningitis in adults . A 20-year overview; Sigurdardottir B et al.; BACKGROUND: Most clinical overviews of acute bacterial meningitis have either focused on children or all age groups combined, although the disease poses serious problems in the adult population . OBJECTIVE: To study the clinical and microbiological features of adult bacterial meningitis in Iceland, as a representative of the average European or North American community . PATIENTS AND METHODS: Data on a total of 132 cases in 127 patients (age, > or = 16 years) who were diagnosed as having acute bacterial meningitis in Iceland during the years 1975 to 1994 were collected from patient and laboratory records . Complete hospital records were found for 119 of the 132 cases identified . RESULTS: The annual incidence was 1.7/100,000 to 7.2/ 100,000 inhabitants (mean, 3.8/100,000) . The most common causative organisms were Neisseria meningitidis (56%), Streptococcus pneumoniae (20%), Listeria monocytogenes (6%), and Haemophilus influenzae (5%) . Neisseria meningitidis caused 93% of the infections in the 16- to 20-year-old age group, but it caused only 25% of the infections in patients aged 45 years or older . Listeria monocytogenes caused 14% of these cases . Cases of nosocomial and recurrent meningitis were rare . A significant underlying illness or condition was present in 39% of the patients . The mean mortality was 19.7%, and it did not change during the study period . CONCLUSIONS: In a study that involved all adult patients with bacterial meningitis in a single country for 2 decades, meningococci and pneumococci were the most frequent causative agents . However, meningococci were responsible for only one fourth of the cases among adult patients aged 45 years or older, most of these cases were caused by pneumococci and Listeria . Despite modern medical developments, approximately 20% of adult patients with bacterial meningitis died.

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