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