Mem Inst Oswaldo Cruz, 2004 Nov, 99(7), 687 - 9 Epub 2005 Jan 12. Naturally acquired immunity to Haemophilus influenzae type B in healthy Cuban children; Torano Peraza G et al.; We have evaluated the prevalence of antibody to immunogenicity of Haemophilus influenzae type b (Hib) in a group of 4 to 5 years old healthy children, who were too old to be included in the first vaccinated cohort when Hib vaccination begun in Cuba in 1999 . Serum capsular polysaccharide specific IgG antibody concentrations were measured in 974 healthy children, between February and May 2002 . The prevalence of Hib nasopharyngeal carriage was also estimated . The majority of children (99.7%) had more than 1 microg/ml of antibody . The preliminary report of the nasopharyngeal cultures was positive for H . influenzae in 16 children, but in only one was confirmed as Hib after serotyping (0.1% Hib nasopharyngeal carrier) . These results provide evidence that in Cuba the natural active immunity to Hib can be acquired at an early age. Chest, 2005 Jan, 127(1), 197 - 204 Antibody Production Deficiency With Normal IgG Levels in Bronchiectasis of Unknown Etiology; Vendrell M et al.; BACKGROUND: No defined cause of bronchiectasis is currently found in approximately 50% of cases . Bronchiectasis is a common long-term complication in patients with primary hypogammaglobulinemia . STUDY OBJECTIVES: To ascertain whether antibody production deficiency with normal total serum IgG levels is associated with bronchiectasis . DESIGN: Antibody response to a pneumococcal unconjugate vaccine and an Haemophilus influenzae type b conjugate vaccine was prospectively studied in all consecutive adult patients with bronchiectasis of unknown etiology who were assessed in our chest outpatient clinic from January 1994 to October 2001 . Serum-specific antibodies were measured by enzyme-linked immunosorbent assay, and the results were compared with those obtained in a healthy adult control group . Antibody production deficiency was defined as a failure to respond to either vaccine . RESULTS: One hundred seven patients were included in the study (mean age, 46.3 years) . Antibody production deficiency was diagnosed in 12 patients (11%) . A significantly higher incidence of otitis media, lower serum IgG(2) subclass levels, and lower preimmunization antibody levels to Streptococcus pneumoniae and H influenzae type b were observed in patients with antibody production deficiency . The probability of antibody production deficiency in patients with a history of otitis media was 20%, 26% in those with low IgG(2) subclass levels, and 58% in those with both a history of otitis media and low IgG2 subclass levels . CONCLUSIONS: Antibody production deficiency with normal IgG levels may be associated with bronchiectasis, making it advisable to evaluate the antibody response to both the H influenzae and pneumococcal vaccines in patients with bronchiectasis of unknown etiology, particularly in those with a history of otitis media, low IgG(2) subclass levels, and low levels of baseline specific antibodies. Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 449 - 56 Multilocus sequence phylogenetic study of the genus Haemophilus with description of Haemophilus pittmaniae sp . nov; Norskov-Lauritsen N et al.; The phylogeny of human isolates of Haemophilus species was estimated based on partial sequences of four separate housekeeping genes . The clustering of each set of sequences was in accordance with speciation of the strains with few exceptions: of 108 gene fragments examined, only three appeared to have been subject to recombination events across the species barrier . Housekeeping gene similarity supported previous DNA-DNA hybridization data for the genus rather than the phylogeny inferred from 16S rRNA gene sequence comparison . The similarity of sequences of Haemophilus parainfluenzae with those of Haemophilus influenzae suggested preservation of the former species in the genus Haemophilus . Three strains representing a novel taxon were unique with respect to the four investigated gene loci . 16S rRNA gene sequence analysis suggested that this taxon belonged to the Parainfluenzae cluster . DNA-DNA hybridization data supported this generic placement . Nine strains of the novel taxon were available for analysis . They were distinct from representatives of all current species of the genus Haemophilus by conventional phenotypic characterization . Genotypic and phenotypic data show that the strains merit recognition as a novel species of Haemophilus . The name Haemophilus pittmaniae sp . nov . is proposed, with HK 85(T) (=CCUG 48703(T)=NCTC 13334(T)) as the type strain. Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 353 - 62 Reclassification of Pasteurella gallinarum, {Haemophilus} paragallinarum, Pasteurella avium and Pasteurella volantium as Avibacterium gallinarum gen . nov., comb . nov., Avibacterium paragallinarum comb . nov., Avibacterium avium comb . nov . and Avibacterium volantium comb . nov; Blackall PJ et al.; This paper describes a phenotypic and genotypic investigation of the taxonomy of {Haemophilus} paragallinarum, Pasteurella gallinarum, Pasteurella avium and Pasteurella volantium, a major subcluster within the avian 16S rRNA cluster 18 of the family Pasteurellaceae . An extended phenotypic characterization was performed of the type strain of {Haemophilus} paragallinarum, which is NAD-dependent, and eight NAD-independent strains of {Haemophilus} paragallinarum . Complete 16S rRNA gene sequences were obtained for one NAD-independent and four NAD-dependent {Haemophilus} paragallinarum strains . These five sequences along with existing 16S rRNA gene sequences for 11 other taxa within avian 16S rRNA cluster 18 as well as seven other taxa from the Pasteurellaceae were subjected to phylogenetic analysis . The analysis demonstrated that {Haemophilus} paragallinarum, Pasteurella gallinarum, Pasteurella avium and Pasteurella volantium formed a monophyletic group with a minimum of 96.8 % sequence similarity . This group can also be separated by phenotypic testing from all other recognized and named taxa within the Pasteurellaceae . As both genotypic and phenotypic testing support the separate and distinct nature of this subcluster, the transfer is proposed of Pasteurella gallinarum, {Haemophilus} paragallinarum, Pasteurella avium and Pasteurella volantium to a new genus Avibacterium as Avibacterium gallinarum gen . nov., comb . nov., Avibacterium paragallinarum comb . nov., Avibacterium avium comb . nov . and Avibacterium volantium comb . nov . The type strains are NCTC 1118(T) (Avibacterium gallinarum), NCTC 11296(T) (Avibacterium paragallinarum), NCTC 11297(T) (Avibacterium avium) and NCTC 3438(T) (Avibacterium volantium) . Key characteristics that separate these four species are catalase activity (absent only in Avibacterium paragallinarum) and production of acid from galactose (negative only in Avibacterium paragallinarum), maltose (negative only in Avibacterium avium) and mannitol (negative in Avibacterium gallinarum and Avibacterium avium). Nucleic Acids Res, 2005 Jan 14, 33(1), 400 - 8 Print 2005. Destabilization of tetranucleotide repeats in Haemophilus influenzae mutants lacking RnaseHI or the Klenow domain of PolI; Bayliss CD et al.; A feature of Haemophilus influenzae genomes is the presence of several loci containing tracts of six or more identical tetranucleotide repeat units . These repeat tracts are unstable and mediate high frequency, reversible alterations in the expression of surface antigens . This process, termed phase variation (PV), enables H.influenzae to rapidly adapt to fluctuations in the host environment . Perturbation of lagging strand DNA synthesis is known to destabilize simple sequence repeats in yeast and Escherichia coli . By using a chromosomally located reporter construct, we demonstrated that the mutation of an H.influenzae rnhA (encoding RnaseHI) homologue increases the mutation rates of tetranucleotide repeats approximately 3-fold . Additionally, deletion of the Klenow domain of DNA polymerase I (PolI) resulted in a approximately 35-fold increase in tetranucleotide repeat-mediated PV rates . Deletion of the PolI 5'>3' exonuclease domain appears to be lethal . The phenotypes of these mutants suggest that delayed or mutagenic Okazaki fragment processing destabilizes H.influenzae tetranucleotide repeat tracts. Nucleic Acids Res . 2005 Jan 13;33(1):e6. Detection and characterization of horizontal transfers in prokaryotes using genomic signature; Dufraigne C et al.; Horizontal DNA transfer is an important factor of evolution and participates in biological diversity . Unfortunately, the location and length of horizontal transfers (HTs) are known for very few species . The usage of short oligonucleotides in a sequence (the so-called genomic signature) has been shown to be species-specific even in DNA fragments as short as 1 kb . The genomic signature is therefore proposed as a tool to detect HTs . Since DNA transfers originate from species with a signature different from those of the recipient species, the analysis of local variations of signature along recipient genome may allow for detecting exogenous DNA . The strategy consists in (i) scanning the genome with a sliding window, and calculating the corresponding local signature (ii) evaluating its deviation from the signature of the whole genome and (iii) looking for similar signatures in a database of genomic signatures . A total of 22 prokaryote genomes are analyzed in this way . It has been observed that atypical regions make up approximately 6% of each genome on the average . Most of the claimed HTs as well as new ones are detected . The origin of putative DNA transfers is looked for among approximately 12 000 species . Donor species are proposed and sometimes strongly suggested, considering similarity of signatures . Among the species studied, Bacillus subtilis, Haemophilus Influenzae and Escherichia coli are investigated by many authors and give the opportunity to perform a thorough comparison of most of the bioinformatics methods used to detect HTs. Vaccine, 2005 Jan 26, 23(10), 1294 - 300 A recombinant P4 protein of Haemophilus influenzae induces specific immune responses biologically active against nasopharyngeal colonization in mice after intranasal immunization; Hotomi M et al.; Outer membrane protein P4, together with P6, is highly conserved among all typeable and nontypeable strains of Haemophilus influenzae (H . influenzae) . Thus, the protein is an attractive antigen for the inclusion in a vaccine against nontypeable H . influenzae (NTHi) . However, the ability of P4 to induce antibodies protective against NTHi infections is still controversial . In this study, we investigated the specific mucosal immune responses against NTHi induced by intranasal immunization with the lipidated form of recombinant P4 protein (rP4) and non-fatty acylated recombinant P6 protein (rP6) with or without cholera toxin (CT) in BALB/c mice model . Intranasal immunization with either rP4+CT, a mixture of rP4 and rP6+CT, or rP4 and rP6 without CT elicited anti-rP4 specific IgG antibody in serum of mice . Intranasal immunization with either rP4+CT or a mixture of rP4, rP6+CT elicited anti-rP4 specific IgA antibody in nasopharyngeal washing (NPW), while intranasal immunization with rP4 and rP6 without CT did not induced anti-rP4 specific IgA antibody responses in NPWs . Sera from mice intranasally immunized with rP4+CT and a mixture of rP4, rP6+CT also showed bactericidal activity . Significant clearance of NTHi in nasopharynx was seen 3 days after the inoculation of live NTHi in mice intranasally immunized with rP4+CT . The current findings suggested that P4 would be a useful antigen as the component of the vaccine to induce protective immune responses against NTHi . The use of an intranasal vaccine composed of the different surface protein antigens is an attractive strategy for the development of a vaccine against NTHi. Microb Pathog, 2005 Jan, 38(1), 23 - 32 Epub 2004 Dec 19. Bovine platelets activated by Haemophilus somnus and its LOS induce apoptosis in bovine endothelial cells; Kuckleburg CJ et al.; Haemophilus somnus is a bacterial pathogen that causes respiratory disease and vasculitis in cattle . Thrombotic meningoencephalitis (TME) and other severe forms of H . somnus-mediated vascular disease are characterized histopathologically by vasculitis, thrombosis, and infiltration of polymorphonuclear cells . It has been reported previously that activated human platelets express CD40L, FasL and P-selectin (CD62P) . We hypothesized that if these surface markers are up-regulated on bovine platelets after in vitro exposure to H . somnus and its lipooligosaccharide (LOS), they might contribute to endothelial cell damage . Using flow cytometry, we demonstrated low baseline expression of these molecules by bovine platelets and increased expression following in vitro stimulation with ADP, H . somnus or H . somnus LOS . H . somnus stimulated platelets were capable of causing apoptosis in endothelial cells as measured by Hoechst-33342 staining and caspase-3 activity . If these events occur in vivo, they might promote vascular damage and endothelial cell apoptosis, leading to the development of vasculitis and thrombosis that characterize bovine H . somnus infection. J Antimicrob Chemother . 2005 Jan 13; {Epub ahead of print} Antimicrobial susceptibility of community-acquired respiratory tract pathogens in the UK during 2002/3 determined locally and centrally by BSAC methods; Morrissey I et al.; To determine the antimicrobial susceptibility of Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae causing community-acquired lower respiratory tract infection in the UK during 2002/2003 and to compare susceptibilities determined locally by disc diffusion with agar dilution MICs determined at a central laboratory . H . influenzae, M . catarrhalis and S . pneumoniae were isolated in 30 laboratories and susceptibility determined locally by the BSAC standardized disc diffusion method . At a central laboratory, isolates were re-identified, tested for beta-lactamase production (H . influenzae and M . catarrhalis only) and MICs determined using the BSAC agar dilution method . Five hundred and eighty-one H . influenzae, 269 M . catarrhalis and 519 S . pneumoniae were collected . Over 93% of M . catarrhalis and nearly 15% of H . influenzae were beta-lactamase positive rendering these sub-populations resistant to aminopenicillins . Overall, the antibacterial susceptibility rates for the isolates were high . However, macrolides showed poor activity against H . influenzae (0.86-1.38% susceptible by disc or MIC methods) and, compared with other antimicrobials, against S . pneumoniae (approximately 88% susceptible) . Between 84% and 95% of H . influenzae, M . catarrhalis and S . pneumoniae were susceptible to cefuroxime but all isolates were susceptible to cefotaxime . Eighty-five percent of H . influenzae were susceptible to trimethoprim . The fluoroquinolones were very active against the isolates, with moxifloxacin showing lower MICs than levofloxacin against S . pneumoniae . Susceptibility determined locally by disc diffusion was in general agreement with that determined centrally by agar dilution MIC testing . However, there was one inconsistency with H . influenzae where disc diffusion indicated 22.9% and 46.8% resistance to clarithromycin and erythromycin, respectively but by MIC, only 0.9% and 6.9% were resistant, respectively . Rates of resistance within community-acquired respiratory tract isolates were relatively low in the UK, in agreement with other studies . Moxifloxacin was the only antibacterial with over 99% isolates susceptible for each of the three pathogens investigated where breakpoints are available . The comparison between disc susceptibility testing and MIC determination using BSAC methods indicated generally good correlation but has highlighted a methodological problem with macrolides against H . influenzae in particular. Lancet, 2005 Jan 1, 365(9453), 43 - 52 Incidences of vaccine-preventable Haemophilus influenzae type b pneumonia and meningitis in Indonesian children: hamlet-randomised vaccine-probe trial; Gessner BD et al.; BACKGROUND: Most studies of Haemophilus influenzae type b (Hib) disease in Asia have found low rates, and few Asian countries use Hib vaccine in routine immunisation programmes . Whether Hib disease truly is rare or whether many cases remain undetected is unclear . METHODS: To estimate incidences of vaccine-preventable Hib pneumonia and meningitis among children younger than 2 years in Lombok, Indonesia, during 1998-2002, we undertook a hamlet-randomised, controlled, double-blind vaccine-probe study (818 hamlets) . Children were immunised (WHO schedule) with diphtheria, tetanus, pertussis (DTP) or DTP-PRP-T (Hib conjugate) vaccine . Vaccine-preventable disease incidences were calculated as the difference in rates of clinical outcomes between DTP and DTP-PRP-T groups . Analyses included all children who received at least one vaccine dose . FINDINGS: We enrolled 55073 children: 28147 were assigned DTP-PRP-T and 26926 DTP . The proportion of pneumonia outcomes prevented by vaccine ranged from less than 0 to 4.8% . Calculated incidences of vaccine-preventable Hib disease (per 10(5) child-years of observation) for outcome categories were: substantial alveolar consolidation or effusion, less than zero (-43 {95% CI -185 to 98}); all severe pneumonia, 264 (95% CI less than zero to 629); all clinical pneumonia, 1561 (270 to 2853); confirmed Hib meningitis, 16 (1.4 to 31); meningitis with cerebrospinal-fluid findings consistent with a bacterial aetiology, 67 (22 to 112); and admission for suspected meningitis or presenting to a clinic with convulsions, 158 (42 to 273) . INTERPRETATION: Hib vaccine did not prevent the great majority of pneumonia cases, including those with alveolar consolidation . These results do not support a major role for Hib vaccine in overall pneumonia-prevention programmes . Nevertheless, the study identified high incidences of Hib meningitis and pneumonia; inclusion of Hib vaccine in routine infant immunisation programmes in Asia deserves consideration. Arch Ophthalmol, 2005 Jan, 123(1), 39 - 44 Penetration pharmacokinetics of topically administered 0.5% moxifloxacin ophthalmic solution in human aqueous and vitreous; Hariprasad SM et al.; OBJECTIVE: To investigate the penetration of 0.5% moxifloxacin hydrochloride into the aqueous and vitreous after topical administration in humans . METHODS: A prospective, nonrandomized study of 20 patients scheduled for vitrectomy surgery between September 1 and December 31, 2003 . Aqueous and vitreous samples were obtained and analyzed after topical administration of 0.5% moxifloxacin hydrochloride, every 2 hours (q2h) or every 6 hours (q6h), for 3 days before surgery . Assays were performed using high-performance liquid chromatography . RESULTS: Mean +/- SD moxifloxacin concentrations in the q2h group for the aqueous (n = 9) and vitreous (n = 10) were 2.28 +/- 1.23 and 0.11 +/- 0.05 microg/mL, respectively . Mean +/- SD moxifloxacin concentrations in the q6h group for the aqueous (n = 10) and vitreous (n = 9) were 0.88 +/- 0.88 and 0.06 +/- 0.06 microg/mL, respectively . The minimum inhibitory concentration for 90% of isolates (MIC(90)) was far exceeded in the aqueous for a wide spectrum of key pathogens, whereas it was not exceeded in the vitreous for several organisms . However, the minimum inhibitory concentration for 50% of the isolates was exceeded in the q2h vitreous group for Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae, Bacillus cereus, and other gram-negative pathogens . CONCLUSIONS: The Endophthalmitis Vitrectomy Study revealed that 94.2% of isolates from postoperative endophthalmitis are gram-positive pathogens . Moxifloxacin has a spectrum of coverage that appropriately encompasses the most common organisms in endophthalmitis . The pharmacokinetic findings of this investigation show that relatively high aqueous levels can be achieved after topical administration . Further studies will help define the precise role of 0.5% moxifloxacin ophthalmic solution in the treatment of or prophylaxis against intraocular infections. Yi Chuan, 2003 Mar, 25(2), 189 - 94 {Identification of Streptococcus Species and Haemophilus influenzae by Direct Sequencing of PCR Products from 16S~23S rRNA Itergenic Sacer Rgions.}; Lu XX et al.; To facilitate species level identification of bacteria without the requirement of presumptive identification,the paper describes a rapid identification method of bacteria by amplification and direct sequencing 16S~23S rDNA intergenic spacer regions (ISR) of the pathogens which cause the upper respiratory tract infective disease by Streptococcus and Haemophilus.Three pairs of primer targeting conserved sequences flanking the 3' end of 16S and the 5'end of 23S rRNA were used to amplify 16S~23S rRNA ISR of 7 streptococcus strains and 8 Haemophilus strains.The PCR products were separated by 1% agarose gel electrophoresis and the polymorphisms fragments were purified with the Wizard PCR Min-Prep Kit (Promega) and Protocol-SK131(Sangon).The nucleotide sequences of ISR inserts were determined by using the XEQTM DTCS KitjjTerminator Cycle Sequencing and a CEQTM 2000XL DNA Analysis system (Backman Coulter) automatic DAN sequencer.Then those sequences were compared with known seqnences on the GenBank.The alignment of nucleotide sequence,evolutionary distances and phylogenetic tress were analyzed by software DANMAN version 4.0.The PCR products were showed polymorphism patterns with agarose gel.One band was contained in streptococcus genus.The significant variation was found among the spacer sequences of different species in Streptococcus with the lengths of the spacer varying from 269 to 446bp.All the ISR of the streptococcal species had a tRNA Ala gene in the spacer and the sequence identities varied from 78 to 88% within genera.It was found that some spacer sequence blocks were highly conserved between operons of a genome,whereas the presence of others was variable,three regions showed significant spatial variation.Most of the differences between the sequences came from several bases insertions/deletions and substitutions.There are two major bands in the Haemophilus biotypes(515 and 884bp),the small ISR amplicon contained one tDNA coding for tRNA(Glu).In contrast to the large one contained two tRNA genes coding for tRAN(Ala) and tRNA(Ile).Two regions of repeating motifs with only A or T were present in higher copy numbers between tRAN(Ala) and tRNA(Ile).The phylogenetic trees varied from 97.5 to 98.8%.The PCR and direct sequencing of 16S~23S rRAN ISR were successful in the pathogen species identification. Zh Mikrobiol Epidemiol Immunobiol, 2004 Nov-Dec, (6), 92 - 4 {Etiological structure of respiratory infections in different variants of acute bronchitis}; Bacteremia among children admitted to a rural hospital in Kenya; Centre for Geographic Medicine Research (Coast), Kilifi, Kenya . jberkley@kilifi.mimcom.net BACKGROUND: There are few epidemiologic data on invasive bacterial infections among children in sub-Saharan Africa . We studied every acute pediatric admission to a rural district hospital in Kenya to examine the prevalence, incidence, types, and outcome of community-acquired bacteremia . METHODS: Between August 1998 and July 2002, we cultured blood on admission from 19,339 inpatients and calculated the incidence of bacteremia on the basis of the population served by the hospital . RESULTS: Of a total of 1783 infants who were under 60 days old, 228 had bacteremia (12.8 percent), as did 866 of 14,787 children who were 60 or more days of age (5.9 percent) . Among infants who were under 60 days old, Escherichia coli and group B streptococci predominated among a broad range of isolates (14 percent and 11 percent, respectively) . Among infants who were 60 or more days of age, Streptococcus pneumoniae, nontyphoidal salmonella species, Haemophilus influenzae, and E . coli accounted for more than 70 percent of isolates . The minimal annual incidence of community-acquired bacteremia was estimated at 1457 cases per 100,000 children among infants under a year old, 1080 among children under 2 years, and 505 among children under 5 years . Of all in-hospital deaths, 26 percent were in children with community-acquired bacteremia . Of 308 deaths in children with bacteremia, 103 (33.4 percent) occurred on the day of admission and 217 (70.5 percent) within two days . CONCLUSIONS: Community-acquired bacteremia is a major cause of death among children at a rural sub-Saharan district hospital, a finding that highlights the need for prevention and for overcoming the political and financial barriers to widespread use of existing vaccines for bacterial diseases . J Immunol, 2005 Jan 15, 174(2), 907 - 17 Viral Delivery of an Epitope from Haemophilus influenzae Induces Central Nervous System Autoimmune Disease by Molecular Mimicry; Croxford JL et al.; Multiple sclerosis (MS) is an autoimmune CNS demyelinating disease in which infection may be an important initiating factor . Pathogen-induced cross-activation of autoimmune T cells may occur by molecular mimicry . Infection with wild-type Theiler's murine encephalomyelitis virus induces a late-onset, progressive T cell-mediated demyelinating disease, similar to MS . To determine the potential of virus-induced autoimmunity by molecular mimicry, a nonpathogenic neurotropic Theiler's murine encephalomyelitis virus variant was engineered to encode a mimic peptide from protease IV of Haemophilus influenzae (HI), sharing 6 of 13 aa with the dominant encephalitogenic proteolipid protein (PLP) epitope PLP(139-151) . Infection of SJL mice with the HI mimic-expressing virus induced a rapid-onset, nonprogressive paralytic disease characterized by potent activation of self-reactive PLP(139-151)-specific CD4(+) Th1 responses . In contrast, mice immunized with the HI mimic-peptide in CFA did not develop disease, associated with the failure to induce activation of PLP(139-151)-specific CD4(+) Th1 cells . However, preinfection with the mimic-expressing virus before mimic-peptide immunization led to severe disease . Therefore, infection with a mimic-expressing virus directly initiates organ-specific T cell-mediated autoimmunity, suggesting that pathogen-delivered innate immune signals may play a crucial role in triggering differentiation of pathogenic self-reactive responses . These results have important implications for explaining the pathogenesis of MS and other autoimmune diseases. BMC Bioinformatics . 2005 Jan 5;6(1):2 {Epub ahead of print} Visualization of comparative genomic analyses by BLAST score ratio; Rasko DA et al.; BACKGROUND: The first microbial genome sequence, Haemophilus influenzae, was published in 1995 . Since then, more than 400 microbial genome sequences have been completed or commenced . This massive influx of data provides the opportunity to obtain biological insights through comparative genomics . However few tools are available for this scale of comparative analysis . RESULTS: The BLAST Score Ratio (BSR) approach, implemented in a Perl script, classifies all putative peptides within three genomes using a measure of similarity based on the ratio of BLAST scores . The output of the BSR analysis enables global visualization of the degree of proteome similarity between all three genomes . Additional output enables the genomic synteny (conserved gene order) between each genome pair to be assessed . Furthermore, we extend this synteny analysis by overlaying BSR data as a color dimension, enabling visualization of the degree of similarity of the peptides being compared . CONCLUSIONS: Combining the degree of similarity, synteny and annotation will allow rapid identification of conserved genomic regions as well as a number of common genomic rearrangements such as insertions, deletions and inversions . The script and example visualizations are available at: http://www.microbialgenomics.org/BSR/. Otolaryngol Head Neck Surg, 2005 Jan, 132(1), 37 - 42 Intracranial complications of otitis media: 15 years of experience in 33 patients; Penido Nde O et al.; Objectives Complications in the central nervous system (CNS) from acute otitis media (AOM) and chronic otitis media (COM) are becoming fewer, although they still represent a challenge for early recognition, adequate treatment, and satisfactory results . This retrospective study analyzed clinical data and therapeutic options in 33 patients with intracranial involvement resulting from otitis media . Important clinical features of intracranial complications and the sequence of the most efficient therapeutic maneuvers are discussed . Methods Charts of six patients with AOM and 27 patients with COM associated with CNS complications were analyzed for clinical presentation, imaging, and therapeutic approach . Results Ages ranged from 6 months to 79 years, with no gender predilection . Persistent fever, headache, and purulent otorrhea were the main symptoms . Proteus mirabilis , Enterococcus , and Pseudomonas aeruginosa were the most common microorganisms in COM, and Pneumococus and Haemophilus were the most common microorganisms in AOM . Nineteen patients (58%) presented with more than 1 CNS complication, resulting in a total of 56 complications, including 26 cases of otogenic brain abscess, 21 cases of meningitis, 5 cases of lateral sinus thromboses, two cases of subdural empyemas, 1 case of epidural empyema, and 1 case of meningocele . Surgical interventions included craniotomy and drainage of the abscess (n = 17), open mastoidectomy with abscess drained through the mastoid (n = 10), open mastoidectomy alone (n = 8), and closed mastoidectomy (n = 2) . Twelve patients who underwent craniotomy had subsequent mastoidectomy for recurrent abscess . At the 6-month, 66% of patients presented without sequelae, 24% presented with sequelae, and 9% died . Conclusion Early identification and prompt clinical and surgical intervention with mandatory drainage of the middle ear (primary disease), was essential for better outcome. CMAJ, 2005 Jan 4, 172(1), 53 - 6 Invasive Haemophilus influenzae type b infections in vaccinated and unvaccinated children in Canada, 2001-2003; Scheifele D et al.; BACKGROUND: Although vaccination of infants against Haemophilus influenzae type b (Hib) invasive infections is effective and has been routinely available in Canada since 1992, cases of the disease continue to occur . We were interested in determining whether recent cases of Hib infection reflected progressive loss of protection with time since vaccination, increasing nonacceptance of vaccination or a deleterious effect of coadministration of recently introduced vaccines such as those for pneumococcal and meningococcal conjugates and hepatitis B . We report on the causes of Hib infections among vaccinated and unvaccinated children between 2001 and 2003 in Canada . METHODS: Through our established network of 12 pediatric tertiary care hospitals we actively searched for cases in each centre by reviewing daily admissions and laboratory reports, visiting the wards and checking discharge diagnosis codes . Culture-confirmed cases were summarized by nurse monitors using a standardized reporting system . RESULTS: We identified 29 cases during the 3 years: 16 in 2001, 10 in 2002 and 3 in 2003 . Half of the 29 patients had meningitis . Hib infection was more common among children less than 6 months of age (11 cases) and in boys (20 cases) . Two deaths occurred (7% case-fatality ratio) . A total of 20 children had received no or incomplete primary vaccination because of parental refusal (7 cases), because they were too young to have completed the primary series (11 cases, including 1 in which parental refusal was also a factor) or because of delays in completing the primary series (2 cases); the vaccination history was uncertain in the remaining case . Infection despite primary vaccination occurred in 9 children: 2 previously healthy children and 7 who were immunocompromised or who had a predisposing condition . None of the cases identified in 2003 involved children who had received any of the newly introduced vaccines . INTERPRETATION: Invasive Hib infections remain rare in Canada, with most cases occurring in children too young to have completed the primary series . Protection after vaccination appears to extend into later childhood and does not appear to be diminished by coadministration of newer infant vaccines. Protein Sci . 2005 Jan 4; {Epub ahead of print} NMR structure of HI0004, a putative essential gene product from Haemophilus influenzae, and comparison with the X-ray structure of an Aquifex aeolicus homolog; Yeh DC et al.; The solution structure of the 154-residue conserved hypothetical protein HI0004 has been determined using multidimensional heteronuclear NMR spectroscopy . HI0004 has sequence homologs in many organisms ranging from bacteria to humans and is believed to be essential in Haemophilus influenzae, although an exact function has yet to be defined . It has a alpha-beta-alpha sandwich architecture consisting of a central four-stranded beta-sheet with the alpha2-helix packed against one side of the beta-sheet and four alpha-helices (alpha1, alpha3, alpha4, alpha5) on the other side . There is structural homology with the eukaryotic matrix metalloproteases (MMPs), but little sequence similarity except for a conserved region containing three histidines that appears in both the MMPs and throughout the HI0004 family of proteins . The solution structure of HI0004 is compared with the X-ray structure of an Aquifex aeolicus homolog, AQ_1354, which has 36% sequence identity over 148 residues . Despite this level of sequence homology, significant differences exist between the two structures . These differences are described along with possible functional implications of the structures. Zhonghua Er Ke Za Zhi, 2004 Nov, 42(11), 854 - 8 {Serotypes and antibiotics-resistance patterns of 247 strains of Haemophilus Influenzae isolated from children in Hangzhou}; Hua CZ et al.; OBJECTIVE: To investigate the serotypes and antibiotics-resistance patterns of Haemophilus influenzae isolated from children in Hangzhou . METHODS: Isolates were identified with api-NH card . Serotypes were determined with slide agglutination method . The sensitivities of 13 antibiotics against 247 strains of Haemophilus influenzae were determined in vitro with Kirby-Bauer diffusion methods and MICs of ampicillin were determined with E-test . Nitrocefin test was used to detect beta-lactamase . RESULTS: Of the 247 strains isolated from children during the period from August 2001 to July 2002, 153 strains were non-typable, while 94 strains (38.1%) were typable and 90.4% and 1.1% of them belonged to type d and type b, respectively . Higher incidence of typable Haemophilus influenzae was found in male than in female children and the difference was significant (chi(2) = 5.30, P < 0.05), while between upper and lower respiratory tract infected children the difference was not statistically significant (chi(2) = 3.60, P > 0.05) . Forty-one isolates (16.6%) were beta-lactamase-positive and 14 strains could not grow on medium in antibiotics sensitivity test . Of all 233 isolates tested successfully, 85.4% were susceptible to ampicillin, and the sensitivity rate to cefaclor, ceftriaxone, cefotaxime, imipenem, rifampin, clarithromycin, and chloramphenicol were as high as 98.7%, 99.6%, 99.6%, 99.6%, 98.7%, 91.0%, and 90.6%, respectively . All strains were sensitive to amoxicillin/clavulanic acid, ampicillin/sulbactan and ofloxacin, while 107 strains (45.9%) were resistant to trimethoprim-sulfamethoxazole, followed by that of tetracycline (14.6%) . Resistance to ampicillin and trimethoprim-sulfamethoxazole in typable isolates was statistically significantly higher than in non-typable strains . Twenty-six strains (10.5%) were multi-resistant isolates and the multi-resistance rate in beta-lactamase-positive strains were significantly higher than that in beta-lactamase-negative strains (chi(c)(2) = 146.8, P < 0.001) . CONCLUSION: Non-typable Haemophilus influenzae was the most common type in clinical strains isolated from children in Hangzhou, while type d was the overwhelming type and type b was uncommon in typable isolates . Incidence of typable isolates was higher in male than in female children, and it was apt to intergrow with other species of pathogenic bacteria . The proportion of beta-lactamase-positive strains was not high and ampicillin or other beta-lactam actibiotics were still the treatment of choice for infections with Haemophilus influenzae. Drugs, 2005, 65(2), 229 - 55 Advances in pneumococcal vaccines : advantages for infants and children; Bernatoniene J et al.; The introduction of Haemophilus influenzae type b (Hib) vaccine into the universal immunisation schedules of many industrialised countries and the subsequent remarkable decline in the incidence of invasive Hib disease has further highlighted the impact of invasive pneumococcal diseases . Streptococcus pneumoniae is now the leading cause of bacterial meningitis in children in many settings and a leading cause of vaccine-preventable bacterial disease in children worldwide.The currently marketed 23-valent pneumococcal polysaccharide vaccine provides large serotype coverage at a relatively low cost . However, it is not efficacious in young children . Pneumococcal conjugate vaccines (PCVs) are highly effective in preventing invasive disease in infants and young children, with favourable safety and immunogenicity profiles . These vaccines have also shown efficacy in reducing cases of non-invasive disease (i.e . otitis media), nasopharyngeal acquisition of vaccine-specific serotypes of S . pneumoniae, and protection against pneumococcal disease caused by resistant strains . However, PCV contains a limited number of pneumococcal serotypes and, given adequate ecological pressure, replacement disease by non-vaccine serotypes remains a threat, particularly in areas with very high disease burden . Furthermore, although capsular-specific antibodies have been shown to be highly protective, it remains unclear what concentration of these serotype-specific antibodies protect against disease and, more recently, it has become clear that opsonic activity and avidity of these antibodies are more critical determinants of protection than concentration . Therefore, monitoring disease burden and defining immune correlates of protection after widespread use of conjugate vaccines are crucial for the evaluation of these new generation vaccines . Furthermore, a need exists to develop pneumococcal vaccines with lower cost and larger serotype coverage.Development of one or more protein vaccines that might be easier and, thus, less expensive to manufacture, and which might provide protection against multiple serotypes, is in progress . This article reviews the current state of pneumococcal disease and pneumococcal vaccines in clinical use. Arch Pathol Lab Med, 2005 Jan, 129(1), 78 - 81 Validation of a rapid diagnostic strategy for determination of significant bacterial counts in bronchoalveolar lavage samples; Laupland KB et al.; CONTEXT: Bacterial cultures of bronchoscopic samples require 1 to 2 days for results to be available for use in clinical decisions . We developed a rapid diagnostic testing strategy that is highly sensitive for screening bacteria in bronchoalveolar lavage (BAL) samples, with results available within hours of collection . OBJECTIVE: To validate the ability of a bacterial adenosine triphosphate (ATP) assay and routine Gram stain microscopy to detect significant bacterial counts in BAL samples . DESIGN: Four hundred seventy-seven BAL samples from 319 patients suspected of having pneumonia were tested using a rapid diagnostic strategy, consisting of Gram stain and a bacterial ATP assay . Rapid results were compared with quantitative cultures with a positive cutoff of 10(4) CFU/mL or higher . RESULTS: Significant bacterial counts were identified in 107 samples (22%) . The most common etiologic agents were Staphylococcus aureus (25%), Haemophilus influenzae (17%), and Streptococcus pneumoniae (12%) . The rapid test results were false negative in 5 cases (S aureus in 2, both Klebsiella pneumoniae and S aureus in 1, and Stenotrophomonas maltophilia and S pneumoniae in 1 case each) . The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the rapid diagnostic strategy were 95.3%, 54.9%, 37.9%, 97.6%, and 63.9%, respectively . CONCLUSION: A negative result with this rapid diagnostic testing strategy rules out significant bacterial counts in BAL samples with a high degree of certainty and may allow use of narrow-spectrum antimicrobial agents or withholding of empiric antimicrobial therapy in patients suspected of having ventilator-associated pneumonia. Arch Pathol Lab Med, 2005 Jan, 129(1), 100 - 3 Haemophilus influenzae lymphadenopathy in a patient with agammaglobulinemia: clinical-histologic-microbiologic correlation and review of the literature; Black C et al.; Agammaglobulinemia is the most common primary immunodeficiency, with an incidence of approximately 1 in 250,000 males in the United States . These patients are at risk for frequent recurrent infections, which may become fatal if untreated . Patients have increased susceptibility to encapsulated pyogenic bacteria . Haemophilus influenzae is second only to Streptococcus pneumoniae as the bacteria most frequently implicated in infections in these patients . We present a case involving an adolescent boy with X-linked agammaglobulinemia and H influenzae cervical adenopathy, confirmed twice by culture . We correlate the clinical, microbiologic, and histologic findings . Owing to the severity of infections in this population, surgical intervention is more common than in the immunocompetent population . This description may help the pathologist in considering a differential diagnosis when examining a diagnostic lymph node biopsy in these patients. Kansenshogaku Zasshi, 2004 Nov, 78(11), 943 - 51 {Antimicrobial susceptibility testing of clinical isolates of Haemophilus influenzae derived from Japanese children}; Hoshino T et al.; We examined the antimicrobial susceptibility testing of 1,024 Haemophilus influenzae isolates recovered from Japanese children . Percent distribution of beta-lactamase-negative ampicillin (ABPC)--resistant strain (BLNAR; ABPC-MIC > or = 4 microg/ml) was 15.0% . Prevalence of BLNAR increased remarkably from 12.7 to 22.1% in recent 4 years . Furthermore, the frequency of highly resistant strains (ABPC-MIC > or = 8 microg/ml) increased from 28.6 to 56.9% among BLNAR . Decreased susceptibility for cephems was observed in non-BLNAR strains including serotype b strain isolated from the cerebrospinal fluid. Int J Pediatr Otorhinolaryngol, 2005 Jan, 69(1), 69 - 74 Nasopharyngeal aerobic bacterial flora and Staphylococcus aureus nasal carriage in deaf children; Harputluoglu U et al.; OBJECTIVE:: To determine the nasopharyngeal aerobic bacterial flora and Staphylococcus aureus nasal carriage in deaf children and the role of flora in deafness . STUDY DESIGN:: A prospective, controlled study . METHODS:: Nasopharyngeal and nasal swabs were collected from 87 deaf children with acquired etiology at Zonguldak primary school for the deaf and 56 healthy children . The children with genetic base (syndromic or nonsyndromic, familial or sporadic, AD, AR or X-linked recessive), and also with the history of drug exposure, head trauma, birth trauma, prematurity, hyperbilirubinemia and the viral diseases with high fever (like mumps and measles) were excluded from the study . Swabs were inoculated on to a variety of bacteriological culture media, which were then incubated in an appropriate atmosphere . Colonisation of Group A beta hemolytic streptococcus, Streptococcus pneumoniae, Haemophilus influenzae, Neisseria menengitidis, Moraxella catarrhalis and S . aureus in upper respiratory tract were investigated . Antimicrobial susceptibility testing of the isolates were determined according to National committee for clinical laboratory standards (NCCLS) guidelines . RESULTS:: Although, the rates of colonization of the nasopharyngeal aerobic bacteria and nasal S . aureus did not differ significantly between deaf children and normal healthy subjects, less colonization rates were found in deaf children than normal healthy subjects . S . aureus was isolated from 18 (20.7%) deaf children . All S . aureus isolates from deaf children were susceptible to oxacillin . Penicillin susceptibility rate was 22.2% . CONCLUSION:: It is considered that nasopharyngeal and nasal colonizations of deaf children with potentially pathogenic aerobic bacterial flora is not a significant risk factor for acquired infections when compared with healthy children. Pediatr Infect Dis J, 2004 Dec, 23(12), 1142 - 52 Haemophilus influenzae: a significant pathogen in acute otitis media; Leibovitz E et al.; Haemophilus influenzae is a major pathogen in acute otitis media (AOM) causing disease that is not clinically distinguishable from that caused by Streptococcus pneumoniae . AOM caused by H . influenzae is particularly associated with older age and recurrent disease . Antibiotics differ in their ability to eradicate H . influenzae from the middle ear space . In the United States, widespread pneumococcal vaccination has increased the importance of H . influenzae as a major therapeutic challenge in the treatment of AOM. Pediatr Infect Dis J, 2004 Dec, 23(12), 1109 - 15 Immunogenicity and reactogenicity of a three-dose primary vaccination course with a combined diphtheria-tetanus-acellular pertussis-hepatitis B-inactivated polio-haemophilus influenzae type b vaccine coadministered with a meningococcal C conjugate vaccine; Tejedor JC et al.; AIM: To evaluate the immunogenicity and safety of 3 doses of the combined diphtheria-tetanus toxoids-acellular pertussis-hepatitis B virus-inactivated poliovirus/Haemophilus influenzae type b (DTPa-HBV-IPV/Hib) vaccine (Infanrix hexa) when coadministered with a CRM197-conjugated meningococcal C vaccine (Meningitec) at different injection sites during the same visit or during separate visits . METHODS: Healthy infants were randomized in an open randomized multicenter study to receive either the DTPa-HBV-IPV/Hib and meningococcal C conjugate vaccines during the same vaccination visit at 2, 4 and 6 months of age (coadministration group) or the DTPa-HBV-IPV/Hib vaccine at 2, 4 and 6 months of age and the meningococcal C conjugate vaccine at 3, 5 and 7 months of age (separate administration group) . RESULTS: The immunogenicity analysis included 452 infants, 228 in the coadministration group and 224 in the separate administration group . One month after primary vaccination, 99.1% of subjects in both groups achieved anti-polyribosylribitol phosphate antibody concentrations >/=0.15 microg/mL . The vaccine response against pertussis antigens was at least 99.1% in both groups . For all other DTPa-HBV-IPV/Hib vaccine antigens, at least 97.8% of all subjects of both groups were seroprotected . In addition, 99.5% of all subjects had meningococcal C bactericidal antibody titers >/=1/8 and 99.1% >/=1/128 . Coadministration of both vaccines did not result in an increased local or general reactogenicity compared with separate administration . CONCLUSION: Coadministration of the combined DTPa-HBV-IPV/Hib vaccine and the meningococcal C conjugate vaccine during the same vaccination visit was immunogenic and safe. Leuk Lymphoma, 2004 Dec, 45(12), 2451 - 4 Similar humoral immunity parameters in chronic lymphocytic leukemia patients independent of v(h) gene mutation status; Sinisalo M et al.; Chronic lymphocytic leukemia (CLL) is a clonal B-cell disorder, which has recently been divided into 2 subtypes based on the somatic hypermutation status of the immunoglobulin heavy chain (IgV(H)) genes . In patients with unmutated tumor cells the survival time is approximately half of that in mutated cases, but the reason for this difference is poorly understood . Since infections are the major cause of mortality in CLL, we investigated the effect of the mutation status on host immunity and proneness to infections in patients with CLL . As expected, the disease progression seemed to be faster and the disease more advanced (Binet B and C) among unmutated patients than in the mutated ones . Surprisingly, no differences in humoral immunity {immunoglobulin G (IgG), IgM, IgA, IgG subclasses, anti-ABO blood group antibodies and mannan-binding lectin (MBL)} or immune responses (Haemophilus influenzae serotype b conjugate vaccination) were detected between these 2 patient groups . Furthermore, UM-patients were not more prone to infections compared to M-patients, and therapy had no impact on the incidence and pattern of infections in either of the patient groups . The current findings within this patient cohort reveal that the worse outcome in the unmutated subgroup is not caused by more severe defects in immunity and increased susceptibility to infections when compared with the hypermutated group . It is thus conceivable that active immunization procedures such as vaccination can successfully be applied on patients with unmutated IgV(H) gene and advanced disease stage. Med Mal Infect, 2004 Nov, 34(11), 493 - 8 {Prevention and infection in adults patients with hyposplenism}; Chanet V et al.; People with asplenia are at risk for infections due to many causative agents, mainly Streptococcus pneumoniae . Among adults, splenectomy is the most frequent etiology of hyposplenism followed with chronic hematological and connective diseases . Physiopathology of the immunologic impairment due to hyposplenia is multifactorial . Physicians and even patients must be aware of overwhelming sepsis occurring on these conditions . The prognosis of these life-threatening infections is related to the precocity of the treatment onset . These infections, mainly due to S . pneumoniae (50-90% of cases) could be prevented with appropriate precautions . Patients presenting with asplenia must be largely vaccinated against these infectious agents: S . pneumoniae, Haemophilus influenzae b, and possibly Neisseiria meningitidis . Oral phenoxymethylpenicillin seems to be the simplest chemoprophylaxis (despite the global increase of pneumococcal strains with reduced susceptibility) . Duration of treatment following splenectomy is discussed: The French medicine agency (AFSSAPS) recommends a 2-year treatment after surgery and for patients having functional hyposplenism (persistency of Howell-Jolly bodies) and/or associated immunodeficiency . Despite these prevention policies, the patient must be informed of the risk of very severe infection. Med Mal Infect, 2004 Feb, 34(2), 97 - 101 {Epidemiology of Haemophilus influenzae strains identified in 2001 in France, and assessment of their susceptibility to beta-lactams}; Dabernat H et al.; The aim of this study was to describe the epidemiology of H . influenzae strains collected in 2001 at the National Reference Center and to evaluate their susceptibility to beta-lactams . METHODS: The demographic characteristics were recorded for each strain, then were determined their capsular serotyping (slide agglutination with specific antisera), as well as their beta-lactamase production (chromogenic cephalosporin test, Nitrocefin), and their MICs (agar dilution method on Haemophilus Test Medium) for amoxicillin (AMX), co-amoxiclav (AMC), cefpodoxime (CPD), cefaclor (CEC), cefuroxime (CXM), and cefotaxime (CTX) . RESULTS: 41.3% of the 752 strains were identified in bronchial secretions, 20.6% in conjunctivitis, 11.3% in otitis media, and 11% in blood cultures . 96.3% of the strains were not capsulated and 3.7% were of type b, d, e or f . 33.8% of the strains were beta-lactamase producers (TEM type), 45.8% of these were identified in otitis pus and 27.7% in bronchial secretions . One hundred and forty-two strains (18.9%) presented reduced susceptibility to beta-lactams (modification of target) associated or not with bla+ . MICs 50/90 against bla+ strains were: AMX 1/32, AMC 0.12/1, CTX 0.007/0.03, CPD 0.03/0.12, CEC 1/64, CXM 0.25/1 . Against low BLNAR and bla+ strains, MICs 50/90 were: AMX 2/32, AMC 0.25/2, CTX 0.015/0.06, CPD 0.06/0.25, CEC 4/64, CXM 0.25/4 . And against low BLNAR strains MICs 50/90 were: AMX 0.25/8, AMC 0.25/8, CTX 0.015/0.12, CPD 0.06/0.50, CEC 4/32, CXM 0.25/4 . CONCLUSIONS: Both bla+ and modifications of PBP are widespread among strains isolated in France . CTX, and CPD remain the most active compounds whatever the resistance mechanisms. Infect Immun, 2005 Jan, 73(1), 609 - 11 Development of a chinchilla model to allow direct, continuous, biophotonic imaging of bioluminescent nontypeable Haemophilus influenzae during experimental otitis media; Novotny LA et al.; We transformed a nontypeable Haemophilus influenzae clinical isolate with a plasmid containing the luxCDABE operon driven by the H . influenzae outer membrane protein P2 promoter . Herein, we demonstrate the ability to detect bioluminescence and to monitor infection within the nasopharynges, eustachian tubes, and middle ears of chinchillas after intranasal and transbullar challenges. Infect Immun, 2005 Jan, 73(1), 599 - 608 A mutation in the sap operon attenuates survival of nontypeable Haemophilus influenzae in a chinchilla model of otitis media; Mason KM et al.; Bacteria have evolved strategies to resist killing by antimicrobial peptides (APs), important effectors of innate immunity . The sap (sensitivity to antimicrobial peptides) operon confers resistance to AP-mediated killing of Salmonella . We have recently shown that sapA gene expression is upregulated in the middle ear in a chinchilla model of nontypeable Haemophilus influenzae (NTHI)-induced otitis media . Based on these findings, we constructed an NTHI strain containing a Lux reporter plasmid driven by the sapA promoter and demonstrated early yet transient expression of the sap operon within sites of the chinchilla upper airway upon infection . We hypothesized that the sap operon products mediate NTHI resistance to APs . In order to test this hypothesis, we constructed a nonpolar mutation in the sapA gene of NTHI strain 86-028NP, a low-passage-number clinical isolate . The sapA mutant was approximately eightfold more sensitive than the parent strain to killing by recombinant chinchilla beta-defensin 1 . We then assessed the ability of this mutant to both colonize and cause otitis media in chinchillas . The sapA mutant was significantly attenuated compared to the parent strain in its ability to survive in both the nasopharynx and the middle ear of the chinchilla . In addition, the mutant was impaired in its ability to compete with the parent strain in a dual-strain challenge model of infection . Our results indicate that the products of the sap operon are important for resisting the activity of APs and may regulate, in part, the balance between normal carriage and disease caused by NTHI. Infect Immun, 2005 Jan, 73(1), 342 - 51 The cytolethal distending toxin induces receptor activator of NF-kappaB ligand expression in human gingival fibroblasts and periodontal ligament cells; Belibasakis GN et al.; Actinobacillus actinomycetemcomitans is associated with localized aggressive periodontitis, a disease characterized by rapid loss of the alveolar bone surrounding the teeth . Receptor activator of NF-kappaB Ligand (RANKL) and osteoprotegerin (OPG) are two molecules that regulate osteoclast formation and bone resorption . RANKL induces osteoclast differentiation and activation, whereas OPG blocks this process by acting as a decoy receptor for RANKL . The purpose of this study was to investigate the effect of A . actinomycetemcomitans on the expression of RANKL and OPG in human gingival fibroblasts and periodontal ligament cells . RANKL mRNA expression was induced in both cell types challenged by A . actinomycetemcomitans extract, whereas OPG mRNA expression remained unaffected . Cell surface RANKL protein was also induced by A . actinomycetemcomitans, whereas there was no change in OPG protein secretion . A cytolethal distending toxin (Cdt) gene-knockout strain of A . actinomycetemcomitans did not induce RANKL expression, in contrast to its wild-type strain . Purified Cdt from Haemophilus ducreyi alone, or in combination with extract from the A . actinomycetemcomitans cdt mutant strain, induced RANKL expression . Pretreatment of A . actinomycetemcomitans wild-type extract with Cdt antiserum abolished RANKL expression . In conclusion, A . actinomycetemcomitans induces RANKL expression in periodontal connective tissue cells . Cdt is crucial for this induction and may therefore be involved in the pathological bone resorption during the process of localized aggressive periodontitis. Antimicrob Agents Chemother, 2005 Jan, 49(1), 309 - 15 In vitro activities of novel 2-fluoro-naphthyridine-containing ketolides; Abbanat D et al.; In vitro activities of erythromycin A, telithromycin, and two investigational ketolides, JNJ-17155437 and JNJ-17155528, were evaluated against clinical bacterial strains, including selected common respiratory tract pathogens . Against 46 macrolide-susceptible and -resistant Streptococcus pneumoniae strains, the MIC(90) (MIC at which 90% of the isolates tested were inhibited) of the investigational ketolides was 0.25 microg/ml, twofold lower than that of telithromycin and at least 64-fold lower than that of erythromycin A . Against erm(B)-containing pneumococci, the MIC(90) of all the ketolides was 0.06 microg/ml . The MIC(90) of the investigational ketolides against mef(A)-containing pneumococci or pneumococci with both mef(A) and erm(B) was 0.25 microg/ml, two-and fourfold lower, respectively, than that of telithromycin . In contrast, the MICs of the investigational ketolides against macrolide-resistant S . pneumoniae strains with ribosomal mutations were similar to or, in some cases, as much as eightfold higher than those of telithromycin . Against Haemophilus influenzae, MICs of all the ketolides were < or =2 microg/ml . Against three Moraxella catarrhalis isolates, the MIC of the ketolides was 0.25 microg/ml . The ketolides inhibited in vitro protein synthesis, with 50% inhibitory concentrations ranging from 0.23 to 0.27 microM . In time-kill studies against macrolide-susceptible and erm- or mef-containing pneumococci, the ketolides were bacteriostatic to slowly bactericidal, with 24-h log(10) decreases ranging from 2.0 to 4.1 CFU . Intervals of postantibiotic effects for the ketolides against macrolide-susceptible and -resistant S . pneumoniae were 3.0 to 8.1 h. Genet Mol Res, 2004 Sep 30, 3(3), 421 - 31 Update of microbial genome programs for bacteria and archaea; Celestino PB et al.; Since the Haemophilus influenzae genome sequence was completed in 1995, 172 other prokaryotic genomes have been completely sequenced, while 508 projects are underway . Besides pathogens, organisms important in several other fields, such as biotechnology and bioremediation, have also been sequenced . Institutions choose the organisms they wish to sequence according to the importance that these species represent to them, the availability of the microbes, and based on the similarity of a species of interest with others that have been sequenced previously . Improvements in sequencing techniques and in associated methodologies have been achieved; however, scientists need to continue working on the development of this field . In Brazil, a multicentered, centrally coordinated and research-focused network was adopted and successfully used for the sequencing of several important organisms . We analyzed the current status of microbial genomes, the trends for criteria used to choose new sequencing projects, the future of microbial sequencing, and the Brazilian genome network. Clin Infect Dis, 2005 Jan 1, 40(1), 17 - 25 Epub 2004 Dec 08. Bacterial meningitis in Burkina Faso: surveillance using field-based polymerase chain reaction testing; Parent du Chatelet I et al.; BACKGROUND: In addition to frequent epidemics of group A meningococcal disease, endemic bacterial meningitis due mostly to Neisseria meningitidis, pneumococcus, and Haemophilus influenzae type b is a serious problem in sub-Saharan Africa . The improved ability to identify the etiologic agent in cases of bacterial meningitis will facilitate more rapid administration of precise therapy . METHODS: To describe the epidemiology of bacterial meningitis and evaluate the usefulness of field-based polymerase chain reaction (PCR) testing, we implemented population-based meningitis surveillance in Burkina Faso during 2002-2003 by use of PCR, culture, and antigen detection tests . RESULTS: Among persons aged 1 month to 67 years, the incidences of meningococcal meningitis, pneumococcal meningitis, and Haemophilus influenzae type b meningitis were 19 cases (n=179), 17 cases (n=162), and 7.1 cases (n=68) per 100,000 persons per year, respectively . Of the cases of meningococcal meningitis, 72% were due to N . meningitidis serogroup W135 . Pneumococcal meningitis caused 61% of deaths and occurred in a seasonal pattern that was similar to that of meningococcal meningitis . Of cases of pneumococcal meningitis and N . meningitidis serogroup W135 meningitis, 71% occurred among persons >2 years of age . Most patients, regardless of the etiology of their illness and the existence of an epidemic, received short-course therapy with oily chloramphenicol . Compared with culture as the gold standard, the sensitivity and specificity of PCR in the field were high; this result was confirmed in Burkina Faso and Paris . CONCLUSIONS: Precise and rapid identification of etiologic agents is critical for improvement in the treatment and prevention of meningitis, and, thus, PCR should be considered for wider use in Africa . Vaccines against Streptococcus pneumoniae, N . meningitidis (including serogroup W135), and H . influenzae type b all will have a major impact on the bacterial meningitis burden . Antibiotic recommendations need to consider the importance of S . pneumoniae, even during the epidemic season. J Infect Chemother, 2004 Dec, 10(6), 352 - 8 Antimicrobial therapy in community-acquired pneumonia among emergency patients in a university hospital in Japan; Kawai S et al.; As antimicrobial therapy for pneumonia has not been well established in Japan, this study was designed to obtain a more definitive standard for antimicrobial treatment of this condition . Two hundred and thirty-one emergency patients admitted to Kyorin University Hospital between January 1998 and December 2000 were retrospectively analyzed in respect to their age, underlying disease, causative organism, and primary treatment with antimicrobial agent . Furthermore, the severity and prognosis were analyzed for those patients who had not responded to initial treatment with antimicrobial agents . The majority of the patients were elderly (over 65 years old; mean overall age 66.7 +/- 15.2 years) and had severe pneumonia; underlying diseases were recognized at a high rate in patients with severe pneumonia (P < 0.05) and in those classified as elderly (P < 0.0001) . The most common underlying conditions in elderly patients were respiratory, cardiovascular (P < 0.01), and cerebrovascular (P < 0.05) diseases . The most common causative organisms were Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae, and Mycoplasma pneumoniae . In patients with severe pneumonia, S . aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa were identified as the most common causative organisms . Complications associated with antimicrobial treatment were observed in those patients with K . pneumoniae isolates who also had severe pneumonia and were frequently treated with penicillin . Furthermore, increased mortality rates were observed in patients not responding well to the initial treatment with antimicrobial agents . Thus, the selection of appropriate initial antimicrobial agents is an important factor affecting the prognosis of patients with community-acquired pneumonia. Paediatr Drugs, 2004, 6(6), 333 - 346 Acute Hematogenous Osteomyelitis in Children : Recognition and Management; Steer AC et al.; Acute hematogenous osteomyelitis is most common in children and has the potential to cause life-long musculoskeletal deformities . Most cases are caused by Staphylococcus aureus . Haemophilus influenzae type b (Hib) is now rare in countries that routinely use the Hib vaccine . Although magnetic resonance imaging is the preferred modality in localized disease, scintigraphy is often preferred as the first line of investigation because it helps to clarify the location of infection and exclude the presence of multifocal disease . Where the presentation is typical, there is no underlying disease, there is a low prevalence of community-acquired methicillin-resistant S . aureus (CA-MRSA), and there is a good response to antibacterial therapy, a diagnostic bone aspirate or biopsy is not necessary . The first-line antibacterial choice in most circumstances is a beta-lactamase-resistant penicillin . If CA-MRSA is suspected, the first-line options include clindamycin, the addition of an aminoglycoside or, rarely, vancomycin . In most patients, the total duration of therapy can be substantially shorter than the traditional 6 weeks, and oral therapy can be commenced after a brief course of intravenous antibacterials . We recommend 3 days of intravenous therapy followed by 3 weeks of high-dose oral antibacterials, provided there is no underlying illness, the presentation is typical and acute, and there has been a good response to treatment initially . Any deviation from this requires more intensive confirmation of the diagnosis (with imaging and/or biopsy or aspiration), and prolongation of intravenous therapy and total duration of treatment . Close monitoring and follow-up for at least 2 years are advised to detect complications. J Health Popul Nutr, 2004 Sep, 22(3), 275 - 85 Evaluation of serogroup A meningococcal vaccines in Africa: a demonstration project; Soriano-Gabarro M et al.; Endemic and epidemic meningococcal disease constitutes a major public-health problem in African countries of the 'meningitis belt' where incidence rates of the disease are many-fold higher (up to 25 cases per 100,000 population) than those in industrialized countries, and epidemics of meningococcal disease occur with rates as high as 1,000 cases per 100,000 people . Using the precedent established during the licensing of conjugate vaccines against Haemophilus influenzae type b and serogroup C meningococci and components of currently-licensed meningococcal polysaccharide vaccines, new meningococcal conjugate vaccines will likely be licensed using immunological endpoints as surrogates for clinical protection . Post-licensure evaluation of vaccine effectiveness will, therefore, be of increased importance . One vaccine being developed is the serogroup A meningococcal (Men A) conjugate vaccine produced by the Meningitis Vaccine Project (MVP), a partnership between the World Health Organization and the Program for Applied Technology in Health . This vaccine will likely be the first meningococcal conjugate vaccine introduced on a large scale in Africa . This paper summarizes the general steps required for vaccine development, reviews the use of immunogenicity criteria as a licensing strategy for new meningococcal vaccines, and discusses plans for evaluating the impact of a meningococcal A conjugate vaccine in Africa . Impact of this vaccine will be measured during a vaccine-demonstration project that will primarily measure the effectiveness of vaccine . Other studies will include evaluations of safety, vaccine coverage, impact on carriage and herd immunity, and prevention-effectiveness studies. J Health Popul Nutr, 2004 Sep, 22(3), 257 - 67 Use of vaccine trials to estimate burden of disease; Mulholland EK; Vaccine trials, the most informative way of determining the efficacy of a vaccine, can also provide valuable information about the burden of disease . The burden of Haemophilus influenzae type b (Hib) remains a major barrier to the use of Hib vaccines, especially in Asia . Recent studies in Indonesia and Bangladesh have used vaccine-trial designs, with known effective vaccines, to estimate the vaccine-preventable burden of Hib disease in those communities . New vaccines against pneumonia and diarrhoeal diseases are usually directed at only one of various causes of the syndrome . In the case of pneumonia, it is very difficult to determine the aetiology in most cases, so the vaccine trial offers a means of determining the burden of vaccine-preventable diseases . This is particularly important for pneumococcal vaccines as serotype replacement may reduce the effectiveness of the vaccines in the field . This approach would underestimate disease burden if vaccines were found to have an impact on syndromes other than those against which they are directed, and might lead to errors in estimation if there were erroneous assumptions about the efficacy of the vaccine against the condition under investigation. J Health Popul Nutr, 2004 Sep, 22(3), 246 - 56 Introducing new vaccines in developing countries: concepts and approaches to estimating burden of Haemophilus influenzae type b-associated disease; Kilgore PE et al.; In the past 30 years, great strides have been made in immunizing infants and children routinely in developing countries under the Expanded Programme on Immunization . Despite this, the introduction of Haemophilus influenzae type b (Hib) vaccines has progressed rather slowly compared to previously-introduced vaccines for infant immunizations . This slower uptake has been attributed partly to the need for data on the burden of invasive Hib disease . To understand this need, conceptual underpinnings and prerequisites were explored for Hib disease-burden studies . Methodological approaches were also reviewed for conducting Hib disease-burden studies that may be considered in developing countries . Potential studies span a range of designs that provide varying levels of clinical, laboratory and epidemiologic evidence of the burden of invasive Hib disease . Carefully-conducted studies can lay the foundation for complementary studies of long-term disability due to invasive Hib disease, national economic analysis, and field evaluations of vaccine . Studies done in collaboration with national agencies and clinical investigators will maximize study value and provide critical data for national decision-makers who make choices regarding the introduction of Hib vaccines. Chemotherapy, 2004 Dec, 50(6), 265 - 75 Epub 2004 Dec 08. In vitro antibacterial activities of new fluoroquinolones against clinical isolates of haemophilus influenzae with ciprofloxacin-resistance-associated alterations in GyrA and ParC; Yoshizumi S et al.; BACKGROUND: The in vitro antimicrobial activities of new fluoroquinolones were tested against quinolone-resistant Haemophilus influenzae of clinical isolates . METHODS: The nucleotide sequences of the gyrA and parC genes from three ciprofloxacin-resistant strains of Haemophilus influenzae (MIC, 1.56-6.25 microg/ml) were determined . The gyrase was purified from the clinical isolates, and the inhibitory activities of quinolones against the enzyme were tested . RESULTS: These strains possessed at least one amino acid substitution in each of the GyrA (asparagine at residue 88 (Asp-88) to Tyr, Ser-84 to Leu or Ser-84 to Leu and Asp-88 to Asn) and ParC (Glu-88 to Lys) . The antibacterial activity of olamufloxacin against the resistant strains was most potent compared with other quinolones, and the inhibitory activities correlated with quinolone resistance of these strains . CONCLUSIONS: These results warrant the clinical effects of new types of fluoroquinolones, such as olamufloxacin, against respiratory tract and otolaryngology infections caused by ciprofloxacin-resistant H . influenzae . 2004 S . Karger AG, Basel. Treat Respir Med, 2004, 3(5), 269 - 77 Short-course therapy for acute sinusitis: how long is enough? Elies W, Huber K. This review examines the issues surrounding short-course antibiotic therapy of acute sinusitis . Acute bacterial sinusitis is a common community-acquired infection defined as inflammation of one or more paranasal sinuses, most often the maxillary sinus . It is estimated that 0.5-5% of colds are complicated by acute sinusitis . Up to 1 in 20 upper respiratory tract infections is complicated by bacterial sinusitis, most often caused by Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Staphylococcus aureus . Early diagnosis and appropriate antibiotic therapy, in combination with agents that relieve nasal congestion, are important factors in preventing suppurative complications . Left untreated, it could lead to the development of chronic sinusitis or epidural or subdural empyema, brain abscess, or cavernosus sinus thrombosis . Isolation of the causal organism is often lacking in the community setting . Empiric antibiotic therapy should provide adequate coverage against the most important pathogens . Guidelines from different specialist societies based on current scientific knowledge are helpful in making the decision on which drug to use . Recommendations for duration of treatment of acute sinusitis are inconsistent between different guidelines but usually a 10- to 14-day treatment course is recommended.Recognition that the 10- to 14-day duration of therapy is not derived from a strong scientific or medical rationale has led some clinicians to call for shortening the duration of antibiotic therapy for patients with upper respiratory tract infections.Accumulating evidence suggests that short-course (< or =5 days) antibiotic therapy may have equivalent or superior efficacy compared with traditional longer (10-14 days) therapies and offers a number of advantages . Results of a number of clinical trials investigating 5-day therapy with oral cephalosporins, new quinolones or ketolides in acute (presumed) bacterial sinusitis in comparison with traditional 10-day treatment courses have been published demonstrating equivalent efficacy of 5-day and 10-day regimens.The evidence reviewed in this article strongly supports reduction of the traditional 10-day course of antibacterial therapy to a 5-day course for uncomplicated acute maxillary sinusitis in adults . Further research related to the duration of antibacterial therapy for sinusitis is needed in children and in adult patients with frontal, ethmoidal and sphenoidal sinusitis. Int J Clin Pract, 2004 Nov, 58(11), 1045 - 7 Haemophilus influenzae pericarditis with tamponade as the initial presentation of systemic lupus erythematosus; Yeh YH et al.; Although cardiac tamponade is an important and emergent complication of systemic lupus erythematosus (SLE), purulent pericarditis is rare despite the high frequency of pericardial effusion in SLE . We describe the first SLE case of Haemophilus influenzae type-f pericarditis with cardiac tamponade with SLE as the initial presentation . The pathophysiology and therapy are discussed. Int J Infect Dis, 2005 Jan, 9(1), 27 - 36 PROTEKT 1999-2000: a multicentre study of the antimicrobial susceptibility of respiratory tract pathogens in Japan; Inoue M et al.; DESIGN:: A six-centre study in Japan during the winter of 1999-2000 assessed the in vitro activity of >20 antimicrobial agents against the common respiratory pathogens Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae, and Moraxella catarrhalis . The minimum inhibitory concentrations (MIC) of each antimicrobial was determined against these isolates using National Committee for Clinical Laboratory Standards (NCCLS) methodology . RESULTS:: Among S . pneumoniae isolates, 44.5% were penicillin resistant . The macrolide resistance rate was 77.9% with 90.5% of penicillin-resistant strains also being macrolide resistant . Resistance mechanisms in macrolide-resistant isolates were identified as mef(A) or erm(B) in 42.5% and 52.5%, respectively . Of the fluoroquinolone-resistant isolates (1.3%), most were also penicillin and macrolide resistant . All strains were inhibited by telithromycin at </=1mg/L . Among S . pyogenes isolates, erythromycin resistance was 17.5% overall but showed considerable variation among the six centres . For H . influenzae, 8.5% produced beta-lactamase and a single beta-lactamase-negative, ampicillin-resistant isolate (0.36%) was obtained, and there was no fluoroquinolone resistance . All isolates were susceptible to telithromycin . Most antimicrobials showed good activity against M . catarrhalis, although 96.7% were beta-lactamase positive . CONCLUSION:: The prevalence of antimicrobial resistance to macrolides, penicillin and the fluoroquinolones among the common respiratory pathogens is high in Japan. Pediatr Infect Dis J, 2004 Oct, 23(10), 944 - 50 Four and one-half-year follow-up of the effectiveness of diphtheria-tetanus toxoids-acellular pertussis/Haemophilus influenzae type b and diphtheria-tetanus toxoids-acellular pertussis-inactivated poliovirus/H . influenzae type b combination vaccines in Germany; Kalies H et al.; BACKGROUND: Recently an increase in the number of invasive Haemophilus influenzae type b (Hib) cases was observed in the United Kingdom, which coincided with a temporary change from diphtheria-tetanus toxoids-wild-type pertussis to diphtheria-tetanus toxoids-acellular pertussis (DTaP) Hib vaccines . A study in Germany based on approximately 2 years of follow-up, estimated vaccine effectiveness (VE) of DTaP/Hib and DTaP-inactivated poliovirus/Hib combination vaccines against invasive Hib disease to be high . OBJECTIVES: To assess VE of DTaP-containing Hib vaccines against Hib in Germany with the use of extended follow-up of case surveillance and vaccine uptake . SUBJECTS AND METHODS: Cases with confirmed systemic Hib infections in children born between June 1, 1996 and December 31, 1998 were ascertained by a nationwide active surveillance system from January 1998 through June 2002 . A representative subcohort of 667 children born in the same time frame was randomly sampled in a nationwide vaccine coverage survey . VE was determined with a case-cohort approach of Cox regression with time-dependent covariates . RESULTS: Thirty-six cases of Hib disease were reported . Of these, 10 were vaccinated with DTaP-containing Hib vaccines only and 20 were not vaccinated . Of the 10 vaccinated cases, 4 had received an incomplete primary series (1-2 doses), and 6 had received the full primary series (3 doses), 3 of whom also received the booster dose . VE of combination vaccines against invasive Hib infection was 89.6% {95% confidence interval (CI), 67.0-96.7} for an incomplete primary series, 96.7% (95% CI 87.7-99.1) for a full primary series and 98.5% (95% CI 94.5-99.6) for a booster dose (irrespective of priming) . CONCLUSION: Hib combination vaccines containing acellular pertussis antigens continue to be highly effective in Germany. J Trop Pediatr . 2004 Dec 15; {Epub ahead of print} Long-term Outcome for Children with Bacterial Meningitis in Rural Papua New Guinea; Wandi F et al.; This study was undertaken to evaluate the long-term neurological outcome for survivors of bacterial meningitis in rural Papua New Guinea . Children who were discharged from Nonga Base Hospital in Rabaul with a diagnosis of bacterial meningitis between 1992 and 2000 were evaluated in their home villages or on review at hospital . Neurological and developmental complications were documented . The outcomes for 80 of 121 eligible children were determined; eight had died following hospital discharge and 41 were lost to follow-up . Major neurological sequalae were found in 50 (63 per cent) of surviving children, and 27 (34 per cent) had multiple severe complications . In rural Papua New Guinea meningitis causes high rates of mortality and severe long-term disability in a high proportion of survivors . High-level resistance to chloramphenicol is likely to be part of the reason for this, but widespread availability of third-generation cephalosporins for the treatment of meningitis, although urgently required, will not overcome the other problems of delayed presentation with established complications . There is a need for the introduction of conjugate Haemophilus influenzae vaccine, and affordable vaccination strategies against Streptococcus pneumoniae . Richer countries could sponsor these vaccines in developing countries, and apply pressure on vaccine producers to lower the costs. J Infect Dis, 2005 Jan 1, 191(1), 58 - 64 Epub 2004 Dec 01. Suppression and modulation of cellular and humoral immune responses to Haemophilus influenzae type B (Hib) conjugate vaccine in hib-diphtheria-tetanus toxoids-acellular pertussis combination vaccines: a study in a rat model; Mawas F et al.; We assessed a rat model to evaluate the immunogenicity of Haemophilus influenzae type b (Hib) conjugate vaccines and the effect on Hib immunogenicity of combining 2 Hib vaccines (Hib-tetanus toxoid {TT}-A and Hib-TT-B) with diphtheria-TT-acellular pertussis (DTaP)(3) or DTaP(5)/inactivated poliovirus (IPV) vaccines . Rats were immunized subcutaneously with Hib alone or with Hib and DTaP-based vaccines; anti-Hib capsular polysaccharide IgG, poly-ribosyl-ribitol-phosphate (PRP), IgG subclass, and cellular immune responses were evaluated . Results showed a significant reduction in the antibody response to PRP when Hib-TT-A was administered in combination with DTaP(3) and showed changes in the anti-PRP IgG subclass distribution between the separate and combination groups . However, combining Hib-TT-B with DTaP(5)/IPV did not reduce the anti-PRP antibody response . These results suggest that the model can predict the effect of combined administration of Hib and DTaP vaccines on Hib immunogenicity and would be suitable for preclinical studies of mechanisms of interference in Hib/DTaP vaccines. Indian J Med Res, 2004 Nov, 120(5), 489 - 94 Antimicrobial susceptibility of bacterial pathogens in the oropharynx of healthy school children in Turkey; Gazi H et al.; BACKGROUND & OBJECTIVES: Information on oropharyngeal carriage rates of Streptococcus pneumoniae, Haemophilus influenzae, Streptococcus pyogenes and Moraxella catarrhalis and their resistance pattern in healthy school children in Turkey is lacking . The present study was undertaken to determine the carriage rates and antimicrobial resistance of these bacterial pathogens in such children aged 6-14 yr in Manisa, Turkey . METHODS: A total of 1022 children were included from nine schools selected randomly from 32 schools . Throat swabs were cultured for bacteria which were identified using standard microbiological methods . Antimicrobial susceptibility was determined as per National Committee for Clinical Laboratory Standards guidelines . RESULTS: Of the 1022 children 240 (23.4%) harboured S . pneumoniae, 162 (15.8%) H . influenzae, 30 (2.9%) S . pyogenes and 82 (8%) M . catarrhalis in their oropharynx . For S . pneumoniae overall 17.9 per cent of the isolates were intermediately and 7 per cent were resistant to penicillin and resistance to erythromycin trimethoprim-sulphamethoxasole (TMP/SMX), and chloramphenicol was 13.7, 9.1 and 1.6 per cent, respectively . Ampicillin resistance observed in 20.9 per cent of H . influenzae isolates was associated with the presence of beta-lactamase, except two isolates interpreted as beta-lactamase-negative ampicillin resistant strains . Resistance of H . influenzae to TMP/SMX, chloramphenicol, azithromycin, cefaclor and amoxicillin/clavulanic acid was 14.2, 2.4, 1.8, 1.2 and 1.2 per cent, respectively . M.catarrhalis isolates produced beta-lactamase in 80.5 per cent of the cases and all were susceptible to macrolides and clavulanic acid/amoxicillin combination; the highest rate of resistance of 17 per cent was for TMP/SMX . One (3.3%) isolate of S . pyogenes was resistant to macrolides tested . INTERPRETATION & CONCLUSION: Our data shows that upper respiratory tract of about 50 per cent children was colonized with respiratory pathogens . There is a need for surveillance of nasopharyngeal carriage of resistant strains in healthy school children. J Med Microbiol, 2005 Jan, 54(Pt 1), 83 - 5 Antimicrobial resistance in the nasopharyngeal flora of children with acute otitis media and otitis media recurring after amoxicillin therapy; Brook I et al.; The objective of this study was to investigate the antimicrobial susceptibility of the organisms isolated from the nasopharynx of children who presented with acute otitis media (AOM) or otitis media that recurred after amoxicillin therapy . Nasopharyngeal cultures obtained from 72 patients, 40 with AOM and 32 with recurrent otitis media (ROM), were analysed . Thirty-six potentially pathogenic organisms were recovered in 34 (85 %) of the children from the AOM group, and 42 were isolated from 29 (91 %) of the children from the ROM group . The organisms isolated were Streptococcus pneumoniae (n = 26), Haemophilus influenzae non-type b (n = 22), Moraxella catarrhalis (n = 13), Streptococcus pyogenes (n = 8) and Staphylococcus aureus (n = 9) . Resistance to the eight antimicrobial agents used was found in 37 instances in the AOM group as compared to 99 instances in the ROM group (P < 0.005) . The difference between AOM and ROM was significant with Streptococcus pneumoniae resistance to amoxicillin (P < 0.005), to amoxicillin/clavulanate (P < 0.005), to trimethoprim/sulfamethoxazole (P < 0.01), to cefixime (P < 0.01) and to azithromycin (P < 0.01), and for H . influenzae resistance to amoxicillin (P < 0.025) . These data illustrate the higher recovery rate of antimicrobial-resistant Streptococcus pneumoniae and H . influenzae from the nasopharynx of children who had otitis media that recurred after amoxicillin therapy than those with AOM. Curr Eye Res, 2004 Oct-Nov, 29(4-5), 245 - 51 Non-steroidal anti inflammatory agents decrease bacterial colonisation of contact lenses and prevent adhesion to human corneal epithelial cells; Bandara BM et al.; Purpose . To investigate non-steroidal anti-inflammatory agents (NSAIDs), salicylic acid, sodium diclofenac and ketorolac for inhibition of bacterial colonization of contact lenses (CL) and human corneal epithelial cells (HCE) . Methods . CLs pre-colonised with Pseudomonas aeruginosa, Haemophilus influenzae, Staphylococcus epidermidis and Streptococcus pneumoniae were exposed overnight to NSAIDs and the number of viable bacteria on the CLs were calculated . Cytotoxicity of NSAIDs to HCE cells was evaluated with the MTT assay . Viable counts were used to measure the adhesion of P . aeruginosa and S . epidermidis to HCE cells in the presence of the least cytotoxic NSAID . Results . All NSAIDs significantly decreased bacterial colonization of CLs in a dose-dependent manner . Salicylic acid (100 mM) completely inhibited colonisation of all species tested and was the least cytotoxic . Salicylic acid also prevented adhesion of P . aeruginosa and S . epidermidis to HCE (60% and 58% inhibition at 60 mM at 2 hours) . Conclusions . Salicylic acid demonstrated potential as a compound for incorporation into anti-bacterial strategies to prevent bacterial contamination of contact lenses . This study highlighted the potential for NSAIDs as anti-bacterial agents and indicates that this class of compound should be investigated for other suitable candidates. ORL J Otorhinolaryngol Relat Spec, 2004, 66(5), 233 - 40 Genotyping of Streptococcus pneumoniae and Haemophilus influenzae isolated from paired middle ear fluid and nasopharynx by pulsed-field gel electrophoresis; Hotomi M et al.; Twenty-eight isolates of Streptococcus pneumoniae and 30 isolates of Haemophilus influenzae from paired nasopharynx and middle ear fluids of 21 children with acute otitis media (AOM) were evaluated to determine genotypes by polymerase chain reaction and pulsed-field gel electrophoresis (PFGE) . Among the 28 isolates of S . pneumonaie, 21 isolates (75.0%) possessed mutations in the pbp1a,pbp2x, and pbp2b genes, and 7 isolates (25%) had mutations in the pbp2x gene . Nineteen isolates (67.9%) expressed the mefE gene, and 5 isolates (17.9%) possessed the ermB gene . Among the 30 isolates of H . influenzae, 5 isolates (16.7%) had mutations in pbp3 genes, 3 isolates (10.0%) produced beta-lactamase, and 2 (6.7%) isolates possessed mutations both in the pbp3 gene and the beta-lactamase gene . Ten out of the 14 pairs (71.4%) of the restriction fragment patterns of S . pneumoniae from paired nasopharynx and middle ear fluids were indistinguishable following PFGE analysis . The same patterns were identified among 5 children of unrelated families . The restriction fragment patterns of H . influenzae isolated by PFGE were also indistinguishable in 13 out of the 15 pairs (86.7%) of nasopharynx and middle ear fluids . The genetic similarity between nasopharyngeal and middle ear isolates suggests that the causative bacteria migrate from the nasopharynx into the middle ear cavity via the Eustachian tube . Some resistant strains might be prevalent . In children with AOM, the nasopharynx could have been colonized by a virulent strain of bacteria that replaced the benign, commensal bacteria and then progressed to the middle ear, where they caused AOM . Microbiology, 2004 Dec, 150(Pt 12), 3935 - 45 Production and characterization of murine monoclonal antibodies against Haemophilus parasuis and study of their protective role in mice; Tadjine M et al.; Monoclonal antibodies (MAbs) against Haemophilus parasuis were obtained by the fusion of SP2/0-Ag14 murine myeloma cells and spleen cells from BALB/c mice immunized with a whole-bacterial-cell suspension (WC) of H . parasuis strain SW124 (serotype 4) . Two MAbs showing strong reactivity in ELISA were further characterized using SDS-PAGE and Western-blot assays . Different treatments of the WC indicated that MAbs 4D5 and 4G9 identified epitopes of proteinic and polysaccharidic nature, respectively . Electron microscopic examination revealed that, unlike the proteinic epitopes, the lipopolysaccharidic epitopes were exposed on the surface of the cell . Using coagglutination, Western-blot and dot-blot assays it was found that both MAbs recognized common epitopes of all the reference strains and field isolates of H . parasuis . None of the other bacteria tested reacted with the MAbs . These results indicated that both the proteinic and polysaccharidic antigens carried species-specific epitopes . It is suggested that these MAbs may potentially be useful for identification of H . parasuis isolates as well as for developing serological diagnostic tools . MAbs 4D5 and 4G9 were unable to kill H . parasuis in vitro in the presence of complement . However, an enhanced bacterial clearance from blood was observed in mice inoculated with either of the MAbs . Highly significant protection was observed in mice using MAb 4G9 . This is believed to be the first report of MAbs capable of identifying common species-specific antigens of H . parasuis and of their implication in protection against challenge infection in mice. Microbiology, 2004 Dec, 150(Pt 12), 3923 - 33 Identification of a haem-utilization protein (Hup) in Haemophilus influenzae; Morton DJ et al.; Haemophilus influenzae has an absolute growth requirement for a porphyrin source . This growth requirement can be satisfied in vitro by haem, haemoglobin or the haemoglobin-haptoglobin, haem-haemopexin and haem-albumin complexes . A family of proteins, termed the Hgp proteins, which are essential for utilization of the haemoglobin-haptoglobin complex, has previously been identified . A strain lacking the Hgp proteins also has a residual ability to utilize haemoglobin, indicating that additional moieties contribute to haemoglobin utilization . Using a haemoglobin affinity method an approximately 105 kDa protein was isolated . Mutation of the identified gene in an Hgp null background reduced the ability of the mutant strain to utilize haemoglobin in vitro . The mutation also resulted in a reduced ability to utilize haem, haem-haemopexin, haem-albumin and haemoglobin-haptoglobin, thus identifying a general haem-utilization protein (Hup) in Haemophilus influenzae. Virology, 2005 Jan 5, 331(1), 6 - 19 Functional comparison of the transposition core machineries of phage Mu and Haemophilus influenzae Mu-like prophage Hin-Mu reveals interchangeable components; Saariaho AH et al.; Bacteriophage Mu uses DNA transposition for propagation and is a model for transposition studies in general . Recent identification of Mu-like prophages within bacterial genomes offers new material for evolutionary and comparative functional studies . One such prophage, Hin-Mu of Haemophilus influenzae Rd, was studied for its transpositional properties . The components of its transposition core machinery, the encoded transposase (MuA(Hin)) and the transposase binding sites, were evaluated for functional properties by sequence comparisons and DNase I footprinting . Transpositional activity of Hin-Mu was examined by in vitro assays directly assessing the assembly and catalytic function of the transposition core machinery . The Hin-Mu components readily assembled catalytically competent protein-DNA complexes, transpososomes . Thus, Hin-Mu encodes a functional transposase and contains critical transposase binding sites . Despite marked sequence differences, components of the Hin-Mu and Mu transposition core machineries are partially interchangeable, reflecting both conservation and flexibility in the functionally important regions within the transpososome structure. Biochemistry, 2004 Dec 14, 43(49), 15534 - 9 Chemical mechanism of the serine acetyltransferase from Haemophilus influenzae; Johnson CM et al.; The pH dependence of kinetic parameters was determined in both reaction directions to obtain information about the acid-base chemical mechanism of serine acetyltransferase from Haemophilus influenzae (HiSAT) . The maximum rates in both reaction directions, as well as the V/K(serine) and V/K(OAS), decrease at low pH, exhibiting a pK of approximately 7 for a single enzyme residue that must be unprotonated for optimum activity . The pH-independent values of V(1)/E(t), V(1)/K(serine)E(t), V/K(AcCoA)E(t), V(2)/E(t), V(2)/K(OAS)E(t), and V/K(CoA)E(t) are 3300 +/- 180 s(-1), (9.6 +/- 0.4) x 10(5) M(-1) s(-1), 3.3 x 10(6) M(-1) s(-1), 420 +/- 50 s(-1), (2.1 +/- 0.5) x 10(4) M(-1) s(-1), and (4.2 +/- 0.7) x 10(5) M(-1) s(-1), respectively . The K(i) values for the competitive inhibitors glycine and l-cysteine are pH-independent . The solvent deuterium kinetic isotope effects on V and V/K in the direction of serine acetylation are 1.9 +/- 0.2 and 2.5 +/- 0.4, respectively, and the proton inventories are linear for both parameters . Data are consistent with a single proton in flight in the rate-limiting transition state . A general base catalytic mechanism is proposed for the serine acetyltransferase . Once acetyl-CoA and l-serine are bound, an enzymic general base accepts a proton from the l-serine side chain hydroxyl as it undergoes a nucleophilic attack on the carbonyl of acetyl-CoA . The same enzyme residue then functions as a general acid, donating a proton to the sulfur atom of CoASH as the tetrahedral intermediate collapses, generating the products OAS and CoASH . The rate-limiting step in the reaction at limiting l-serine levels is likely formation of the tetrahedral intermediate between serine and acetyl-CoA. Med Oral Patol Oral Cir Bucal, 2004, 9 Suppl, 44 - 51; 37-43 Bacterial endocarditis prophylaxis; Blanco-Carrion A; Bacterial endocarditis (BE) is a disease resulting from the association of morphological alterations of the heart and bacteraemia originating from different sources that at times can be indiscernible (infectious endocarditis) . It is classified on the basis of the morphological alteration involved, depending on the clinical manifestations and course of illness, which varies according to the causative microorganism and host conditions (for example, it is characteristic in I.V . drug users) . The most common microorganisms involved are: Streptococcus viridans (55%), Staphylococcus aureus (30%), Enterococcus (6%) and HACEK bacteria (corresponding to the initials: Haemophilus, Actinobacillus, Cardiobacterium, Eikenella and Kingella), although on occasions it can also be caused by fungi . The oral microbiological flora plays a very important role in the aetiopathogenesis of BE, given that the condition may be of oral or dental origin . This paper will deal with the prevention of said bacteraemia . Prophylaxis will be undertaken using amoxicillin or clindamycin according to action protocols, with special emphasis placed on oral hygiene in patients with structural defects of the heart. J Allergy Clin Immunol, 2004 Dec, 114(6), 1456 - 62 Nuclear factor kappaB essential modulator-deficient child with immunodeficiency yet without anhidrotic ectodermal dysplasia; Niehues T et al.; BACKGROUND: Amorphic mutations in the X-linked nuclear factor kappaB essential modulator ( NEMO ) gene cause Incontinentia pigmenti, which is lethal in hemizygous male patients . Hypomorphic NEMO mutations in male patients lead to anhidrotic ectodermal dysplasia (EDA) with immunodeficiency . OBJECTIVE: To report the clinical features of a child bearing a NEMO mutation who displayed an immunodeficiency without EDA . METHODS: Documentation of clinical care, chart review, standard immunologic and microbiological laboratory techniques, mutation analysis of the NEMO gene . RESULTS: Since the age of 15 months, the patient had Mycobacterium avium disease, beginning with multiple adenitis, later followed by disseminated osteomyelitis and dermatitis . In addition, Haemophilus influenzae and Streptococcus pneumoniae infections led to bronchiectasis . An immunologic work-up revealed a low production of IFN-gamma by PBMCs associated with a hyper-IgM phenotype . Despite treatment using repeated cycles of a 4-drug antimycobacterial regimen, continuous subcutaneous IFN-gamma, repeated antibiotic treatment, and intravenous immunoglobulin substitution, the boy remained chronically ill . At the age of 12 years, the disease was complicated by severe autoimmune hemolytic anemia and eventually fatal herpes simplex virus 1 encephalitis despite high-dose acyclovir therapy . Although he did not present any sign of EDA, a novel type of disease-causing hypomorphic NEMO mutation (110-111insC in exon 2) was identified . CONCLUSION: This case demonstrates that patients hemizygous for NEMO mutations can present with an immunodeficiency without EDA . An investigation of NEMO should thus be undertaken in selected children with immunodeficiency despite the lack of EDA. Pediatr Pulmonol, 2005 Feb, 39(2), 127 - 34 Medical management of parapneumonic pleural disease; Barnes NP et al.; Considerable heterogeneity exists in the management of parapneumonic pleural disease . A randomized controlled trial (RCT) demonstrated the effectiveness of small-catheter drainage with fibrinolysis, but surgical devotees suggest this may only be applicable to "early" cases . We examined evidence-based medical management in "all-comers." We performed a retrospective database analysis of the management of all children with complex pleural effusion admitted to the John Radcliffe Hospital over the 7-year period 1996-2003 . One hundred and ten children were admitted . Ten were excluded as they were part of a multicenter RCT and had received intrapleural saline instead of urokinase . Of the remaining 100, 51 were female and 49 male . Median age on admission was 5.8 years (range, 0.3-16.5) . Symptoms preadmission averaged 11 days, with December the most common month for presentation . Ninety-six underwent chest ultrasound, confirming an effusion in all, described as loculated/septated (68) or echogenic (11) . In 17 cases, no specific comment was made regarding the nature of the fluid seen on ultrasound . Ninety-five had subsequent chest tube drainage and then received intrapleural fibrinolysis with urokinase . An etiological organism was identified in 21 cases (21%) (Streptococcus pneumoniae in 10, group A Streptococcus in 5, Staphylococcus aureus in 4, Haemophilus influenzae in 1, and coliform in 1) . In a further 9 cases (9%), Gram-positive organisms were seen on pleural fluid microscopy, but did not grow on culture . Two (2%) required surgery due to the persistence of symptoms and an inadequate response to medical management . Median duration of admission was 7 days (range, 2-21 days); median duration of stay from intervention was 5 days (range, 2-19 days) . At median follow-up of 8 weeks (range, 3-20 weeks), all children were symptom-free, with minimal pleural thickening on chest X-ray . In conclusion, antibiotic therapy with chest drain insertion and intrapleural urokinase is effective in treating complex parapneumonic effusion and is associated with a good long-term outcome . (c) 2004 Wiley-Liss, Inc. J Oral Maxillofac Surg, 2004 Dec, 62(12), 1545 - 50 Microbiology and management of peritonsillar, retropharyngeal, and parapharyngeal abscesses; Brook I; This review describes the microbiology, diagnosis, and management of peritonsillar, retropharyngeal, and parapharyngeal abscesses in children . Predominant anaerobic organisms isolated in peritonsillar, lateral pharyngeal, and retropharyngeal abscesses are Prevotella, Porphyromonas, Fusobacterium and Peptostreptococcus spp.; aerobic organisms are group A streptococcus ( Streptococcus pyogenes ), Staphylococcus aureus and Haemophilus influenzae . Anaerobic bacteria can be isolated from most abscesses whenever appropriate techniques for their cultivation have been used, while S . pyogenes is isolated in only about one third of cases . More than two thirds of deep neck abscesses contain beta-lactamase producing organisms . Management of tonsillar, peritonsillar, and retropharyngeal abscesses is similar . Systemic antimicrobial therapy should be given in large doses whenever the diagnosis is made . However, when pus is formed, antimicrobial therapy is effective only in conjunction with adequate surgical drainage . Untreated abscesses can rupture spontaneously into the pharynx, causing catastrophic aspiration . Other complications are extension of infection laterally to the side of the neck or dissection into the posterior mediastinum through facial planes and the prevertebral space . Death can occur from aspiration, airway obstruction, erosion into major blood vessels, or extension to the mediastinum. Fitoterapia, 2004 Dec, 75(7-8), 733 - 6 Antibacterial activity of Achillea clavennae essential oil against respiratory tract pathogens; Skocibusic M et al.; The essential oil of Achillea clavennae was investigated for its antibacterial activity against some respiratory tract pathogens . Maximum activity was observed against Klebsiella pneumoniae and penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae . The oil also exhibited strong activity against Gram (-) Haemophilus influenzae and Pseudomonas aeruginosa . Gram (+) Streptococcus pyogenes was the most resistant to the oil. Ann Otol Rhinol Laryngol, 2004 Nov, 113(11), 902 - 5 Effect of antimicrobial therapy with amoxicillin and cefprozil on bacterial interference and beta-lactamase production in the adenoids; Brook I et al.; To compare the effects on the bacterial flora of the adenoids of antimicrobial therapy with amoxicillin or cefprozil, we enrolled in a prospective randomized study 60 children scheduled for elective adenoidectomy because of recurrent otitis media . They were randomized before surgery into 3 groups of 20 patients each: a no-therapy group, and groups with 10 days of either amoxicillin or cefprozil therapy . Core adenoid materials were quantitatively cultured for aerobic and facultative bacteria . The in vitro ability of alpha-hemolytic streptococci (AHS) to inhibit the growth of non-type B Haemophilus influenzae and Streptococcus pneumoniae was determined . The number of organisms in adenoids obtained from patients treated with either antibiotic was reduced as compared to that in adenoids from controls . However, in patients treated with amoxicillin, a significant decline in the number of AHS, and an increase (in Staphylococcus aureus) or no change in the number of other beta-lactamase-producing bacteria (BLPB) was noted . In contrast, in those treated with cefprozil, no change was noted in the frequency of recovery of AHS, and the number of BLPB decreased . Interfering AHS were more often recovered in patients treated with cefprozil . We conclude that the adenoids contain more interfering AHS after therapy with a second-generation oral cephalosporin (cefprozil) than after amoxicillin therapy . This study suggests a potential beneficial effect of using an antimicrobial that selectively spares interfering AHS while inhibiting BLPB. East Mediterr Health J, 2003 Jan-Mar, 9(1-2), 208 - 14 Bone infection; Mousa HA; Osteomyelitis, or bone infection, affects all age groups and develops from various sources including haematogenously from distant infection foci, from external sources such as post-operative or post-traumatic wound infections and from adjoining soft tissue infections . Staphylococcus aureus, Streptococcus pyogenes and Haemophilus influenzae are the most common pathogens of haematogenous osteomyelitis . Aerobic and facultative gram-negative bacteria have emerged as significant pathogens in some types of osteomyelitis while anaerobic bacteria are increasingly recognized as potential pathogens in non-haematogenous osteomyelitis . The emergence of antibiotic resistance is of increasing concern, although improvements in radiologic imaging, antibiotic treatment and heightened awareness have led to earlier detection such that long-term sequelae and morbidity are now primarily due to delays in diagnosis and inadequate treatment. Antimicrob Agents Chemother, 2004 Dec, 48(12), 4766 - 77 Population pharmacokinetics and pharmacodynamics of garenoxacin in patients with community-acquired respiratory tract infections; Van Wart S et al.; Garenoxacin (T-3811ME, BMS-284756) is a novel, broad-spectrum des-F(6) quinolone currently under study for the treatment of community-acquired respiratory tract infections . This analysis assessed garenoxacin population pharmacokinetics and exposure-response relationships for safety (adverse effects {AE}) and antimicrobial activity (clinical cure and bacteriologic eradication of Streptococcus pneumoniae and the grouping of Haemophilus influenzae, Haemophilus parainfluenzae, and Moraxella catarrhalis) . Data were obtained from three phase II clinical trials of garenoxacin administered orally as 400 mg once daily for 5 to 10 days for the treatment of community-acquired pneumonia, acute exacerbation of chronic bronchitis, and sinusitis . Samples were taken from each patient before drug administration, 2 h following administration of the first dose, and on the day 3 to 5 visit . Individual Bayesian estimates of the fu (fraction unbound), the Cmax, and the fu for the area under the concentration-time curve from 0 to 24 h (fu AUC(0-24)) were calculated as measurements of drug exposure by using an ex vivo assessment of average protein binding . Regression analysis was performed to examine the following relationships: treatment-emergent AE incidence and AUC(0-24), Cmax, or patient factors; clinical response or bacterial eradication and drug exposure (fu Cmax/MIC, fu AUC(0-24)/MIC, and other exposure covariates); or disease and patient factors . Garenoxacin pharmacokinetics were described by a one-compartment model with first-order absorption and elimination . Clearance was dependent on creatinine clearance, ideal body weight, age, obesity, and concomitant use of pseudoephedrine . The volume of distribution was dependent on weight and gender . Patients with mild or moderate renal dysfunction had, on average, approximately a 16 or 26% decrease in clearance, respectively, compared to patients of the same gender and obesity classification with normal renal function . AE occurrence was not related to garenoxacin exposure . Overall, clinical cure and bacterial eradication rates were 91 and 90%, respectively, for S . pneumoniae and 93 and 92%, respectively, for the grouping of H . influenzae, H . parainfluenzae, and M . catarrhalis . The fu AUC(0-24)/MIC ratios were high (>90% were >200), and none of the pharmacokinetic-pharmacodynamic exposure measurements indexed to the MIC or other factors were significant predictors of clinical or bacteriologic response . Garenoxacin clearance was primarily related to creatinine clearance and ideal body weight . Although garenoxacin exposure was approximately 25% higher for patients with moderate renal dysfunction, this increase does not appear to be clinically significant as exposures in this patient population were not significant predictors of AE occurrence . Garenoxacin exposures were at the upper end of the exposure-response curves for measurements of antimicrobial activity, suggesting that 400 mg of garenoxacin once daily is a safe and adequate dose for the treatment of the specified community-acquired respiratory tract infections. Antimicrob Agents Chemother, 2004 Dec, 48(12), 4532 - 41 PnuC and the utilization of the nicotinamide riboside analog 3-aminopyridine in Haemophilus influenzae; Sauer E et al.; The utilization pathway for the uptake of NAD and nicotinamide riboside was previously characterized for Haemophilus influenzae . We now report on the cellular location, topology, and substrate specificity of PnuC . pnuC of H . influenzae is only distantly related to pnuC of Escherichia coli and Salmonella enterica serovar Typhimurium . When E . coli PnuC was expressed in an H . influenzae pnuC mutant, it was able to take up only nicotinamide riboside and not nicotinamide mononucleotide . Therefore, we postulated that PnuC transporters in general possess specificity for nicotinamide riboside . Earlier studies showed that 3-aminopyridine derivatives (e.g., 3-aminopyridine adenine dinucleotide) are inhibitory for H . influenzae growth . By testing characterized strains with mutations in the NAD utilization pathway, we show that 3-aminopyridine riboside is inhibitory to H . influenzae and is taken up by the NAD-processing and nicotinamide riboside route . 3-Aminopyridine riboside is utilized effectively in a pnuC+ background . In addition, we demonstrate that 3-aminopyridine adenine dinucleotide resynthesis is produced by NadR . 3-Aminopyridine riboside-resistant H . influenzae isolates were characterized, and mutations in nadR could be detected . We also tested other species of the family Pasteurellaceae, Pasteurella multocida and Actinobacillus actinomycetemcomitans, and found that 3-aminopyridine riboside does not act as a growth inhibitor; hence, 3-aminopyridine riboside represents an anti-infective agent with a very narrow host range. Kansenshogaku Zasshi, 2004 Oct, 78(10), 891 - 7 {The prevalence of beta-lactamase negative ampicillin resistant Haemophilus influenzae in Mie Prefecture}; Maruyama T et al.; Equivalent MIC breakpoints to detect beta-lactamase negative ampicillin resistant Haemophilus influenzae (BLNAR) were controversial . We studied the relationship of drug resistance with gene alterations in 74 clinical isolates of H . influenzae . Out of 74 isolates, 26 showed MIC of ampicillin (ABPC) > or = 1 microg/ml . All isolates, except one, with MIC of ABPC > or = 4 microg/ml were found to produce beta-lactamase, while all 19 isolates with MIC of ABPC at 1 or 2 microg/ml were non-producing . Twenty-six ABPC resistant isolates were subjected to the analysis of genes involved in the drug resistance such as pbp3-1 pbp3-2, and TEM by the Haemophilus influenzae gene detection kit (Wakunaga Pharmaceutical Co., Ltd.) according to the supplier's instructions . Three (21.4%) of 14 beta-lactamase non-producing isolates with ABPC-MIC of 1 microg/ml had mutations of pbp3-1 gene, while all 5 non-producing isolates with ABPC-MIC of 2 microg/ml showed mutations of both pbp3-1 and pbp3-2 genes . A