Haemophilus

Int J Infect Dis, 2003 Jun, 7(2), 143 - 51
Facilitating the WHO expanded program of immunization: the clinical profile of a combined diphtheria, tetanus, pertussis, hepatitis B and Haemophilus influenzae type b vaccine; Aristegui J et al.; BACKGROUND: Vaccines are important weapons in the fight against infectious diseases . The World Health Organization (WHO) Expanded Program on Immunization (EPI) has been extended to include recommendations for hepatitis B and Haemophilus influenzae type b (Hib) vaccinations . The WHO has recommended that combined vaccines be used where possible, to reduce the logistic costs of vaccine delivery . This paper reviews the efficacy, safety and cost-effectiveness of Tritanrix-HB/Hib, the only commercially available combined diphtheria, tetanus, whole cell pertussis, hepatitis B and conjugated Hib vaccine . METHODS: The immunogenicity and reactogenicity results of five published clinical trials involving Tritanrix-HB/Hib in a variety of immunization schedules and countries were reviewed . Based on these data and cost-effectiveness studies, an assessment of its suitability for use in national immunization programs was made . RESULTS: Tritanrix-HB/Hib has shown excellent immunogenicity in clinical trials using a variety of schedules, with no reduced immunogenicity observed for any of the components of the combined vaccine . It has similar reactogenicity to DTPw vaccines alone . Pharmacoeconomic analyses have shown combined DTP-HB/Hib vaccines to be cost-effective compared to separate vaccines . CONCLUSIONS: Replacement of DTPw vaccination by Tritanrix-HB/Hib can be done without modifying the existing national immunization programs . This should facilitate widespread coverage of hepatitis B and Hib vaccinations and their rapid incorporation into the EPI.

Int J Infect Dis, 2003 Jun, 7(2), 109 - 12
Childhood bacterial meningitis in Bulgaria: a population-based retrospective study in six regions during 1992-96; Kojouharova M et al.; OBJECTIVES: We conducted a study to provide information on the importance of bacterial pathogens causing childhood meningitis in Bulgaria . METHODS: A 5-year population-based retrospective survey for bacterial meningitis in children <5 years of age was performed at all hospitals in the six largest regions of Bulgaria . RESULTS: There were 297 cases of meningitis reported, of which 211 (71.0%) were classified as bacterial in origin . The most common causes were Neisseria meningitidis (49 cases) and Haemophilus influenzae type b (Hib) (44 cases), accounting for 36% and 32% of etiologically confirmed cases . Thirty-one cases (70.5%) of Hib meningitis occurred in children <2 years of age, and 26 (59.1%) occurred in children 6-23 months of age . Average annual incidence rates of Hib meningitis based on the population of children <5 years of age for each region ranged from 1.3 to 9.8 per 100,000 (mean 5.9/100,000) . CONCLUSION: The estimated incidence rates from this study were similar to those reported from southern European and Mediterranean countries . Further studies are planned to provide information on appropriate strategies for preventing childhood meningitis in Bulgaria.

Int J Infect Dis, 2003 Mar, 7 Suppl 1, S27 - 31
Implications for antimicrobial prescribing of strategies based on bacterial eradication; Klugman KP; Antimicrobial prescribing in respiratory tract infection is generally empirical . Agents that do not eradicate the key bacterial respiratory pathogens (Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis) provide suboptimal therapy . A recent paper developed by a multidisciplinary, multinational group presented a consensus on the principles that should underpin appropriate antimicrobial prescribing . In summary, in order to ensure clinical success and minimize the threat of resistance, empirical therapy should avoid unnecessary and inappropriate use of antimicrobials, deliver the right agent at the right dose and duration, and rapidly eradicate the pathogen at the site of infection . Accurate diagnosis is essential to ensure that only bacterial infections are treated with antibacterial agents . The application of pharmacokinetic/pharmacodynamic (PK/PD) principles to both new and existing antimicrobials allows the prediction of bacteriologic efficacy . Applying these principles when prescribing therapy can help in reducing the potential for the selection and spread of resistance . Local resistance patterns and the bacteriologic/clinical impact of resistance should also be considered . The use of antimicrobials with optimal PK/PD characteristics may be more cost-effective than allowing the possibility of resistance-induced failure . Changing prescribing habits without taking all these factors into account may increase the incidence of unfavorable patient outcomes and the cost of treatment, with more referrals and hospitalizations . Changes in prescribing habits should be considered carefully, to avoid unintended negative consequences . It is the responsibility of physicians to ensure that each prescription is necessary and will maximize the potential for clinical cure, but there is also a collective responsibility to sustain the diversity of antimicrobial therapy via appropriate formularies, guidelines and licensing, reduced over-the-counter availability, and continued research and development through academia and industry . To maximize clinical cure and minimize the emergence and spread of resistance, antimicrobial prescribing should maximize bacterial eradication, and clinical drug evaluation needs to be brought into line with this need.

Int J Infect Dis, 2003 Mar, 7 Suppl 1, S21 - 6
Achieving bacterial eradication using pharmacokinetic/pharmacodynamic principles; Dagan R; Evidence from studies in otitis media indicates that antimicrobials and dosing regimens that have equivalent bacteriologic efficacy against susceptible pathogens can have significantly different bacteriologic success rates against resistant strains of the same species . Unlike macrolide and fluoroquinolone resistance, penicillin resistance can be overcome in Streptococcus pneumoniae by increasing the dose, and hence increasing the time for which the serum concentrations are above the MIC . The new clinical formulation of extra-strength amoxicillin-clavulanate provides 90 mg/kg per day amoxicillin plus 6.4 mg/kg per day clavulanate (14:1) divided every 12 h, compared with 45/6.4 mg/kg per day b.i.d . with conventional dosing . The pharmacokinetic/pharmacodynamic (PK/PD) profiles of extra-strength amoxicillin-clavulanate predict that the new formulation will be more effective than the conventional formulation against S . pneumoniae with elevated amoxicillin MICs and against Haemophilus influenzae . In an open-label, non-comparative study in children with acute otitis media, the extra-strength formulation had high bacteriologic success rates against the major respiratory pathogens, including penicillin-resistant S . pneumoniae . The development of new antimicrobial agents and formulations should be aimed at meeting PK/PD parameters predictive of bacterial eradication of both susceptible and resistant strains.

Acta Paediatr, 2003 May, 92(5), 541 - 5
Immunogenicity and safety in infants of a DTwPHib full liquid vaccine; Botet Asensi FI et al.; AIM: Combining paediatric vaccines is a rational solution to reduce the number of injections during a single clinical visit, to maintain parents' compliance and to extend vaccine coverage . Different diphtheria, tetanus and whole cell pertussis (DTwP)-containing combination vaccines are licensed and used world-wide . This study assessed the immunogenicity and safety in infants of a combined diphtheria-tetanus-whole cell pertussis-Haemophilus influenzae type b-CRM197 conjugate full liquid vaccine . METHODS: The safety and efficacy of a combined ready-to-use liquid vaccine containing diphtheria and tetanus toxoids, cell suspension of Bordetella pertussis and H . influenzae type b-CRM197 conjugate vaccine (DTwPHib) were assessed in infants eligible for the local Expanded Programme on Immunization (EPI) in Valencia, Spain . The comparative group received separate injections of reference vaccines DTwP + Hib . RESULTS: Local and systemic reactions and adverse events were generally mild and similar in the two groups . DTwPHib elicited anti-PRP antibody titres > or = 0.15 microg ml(-1) in 97% and DTwP + Hib in 94% of infants . Furthermore, 89% of DTwPHib and 78% of DTwP + Hib recipients attained anti-PRP antibody titres > or = 1.0 microg ml(-1), signifying long-term protection . The anti-PRP geometric mean titre was significantly higher in the combined DTwPHib vaccine group (6.65 vs 3.57 microg ml(-1)) . In both groups, 99% of infants achieved protective (> or = 0.01 IU ml(-1)) anti-diphtheria antibody levels and all children achieved protective (> or = 0.1 IU ml(-1)) anti-tetanus antibody levels . DTwPHib caused a > or = 2-fold increase in anti-pertactin antibody titres in 91% and a > or = 4-fold increase in 82% of recipients . The corresponding proportions in the DTwP + Hib group were 95% and 90% . DTwPHib induced a > or = 2-fold increase in anti-Aggl2 and 3 antibody levels in 79% and a > or = 4-fold increase in 73% of recipients . The corresponding proportions among DTwP + Hib infants were 85 and 82% . CONCLUSION: Overall, the combined liquid vaccine DTwPHib is a safe and effective immunogenic vaccine for EPI use in infants.

Mikrobiyol Bul, 2003 Jan, 37(1), 19 - 25
{In vitro sensitivity to antimicrobial agents of Haemophilus influenzae strains isolated from clinical specimens}; Budak F et al.; In-vitro activity of several antimicrobial agents were evaluated against Haemophilus influenzae (n:127) isolated from clinical specimens within the years 2000 and 2002 in a children's hospital . Antimicrobial susceptibility tests were performed by disk diffusion method according to the recommendations of the NCCLS standards and beta-lactamase activity was investigated by nitrocefin test . Resistance rates for the antibiotics were as follows: ampicillin 5.6%, trimethoprim-sulfamethoxazole (TMP/SMX) 27.5%, tetracycline 8.6%, clarithromycin 7%, chloramphenicol 4.7% . Five isolates (3.9%) were found multiple resistant . Beta-lactamase activities were detected in all of the ampicillin resistant isolates . None of the isolates were resistant to ampicillin-sulbactam, cefaclor, cefuroxime, cefprozil, cefpodoksim, cefotaxime, meropenem or ciprofloxacin . According to these results, resistance to ampicillin is low in our H . influenzae isolates compared to other European countries but resistance to TMP/SMX is very high and this finding has to be taken into account in the empirical therapy of community acquired respiratory tract infections.

Bone Marrow Transplant, 2003 Jul, 32(2), 225 - 9
Polysaccharide antibody responses are impaired post bone marrow transplantation for severe combined immunodeficiency, but not other primary immunodeficiencies; Slatter MA et al.; Established treatment of severe combined immunodeficiencies (SCID) and other primary immunodeficiencies (PID) is bone marrow transplantation (BMT) . Normal lymphocyte numbers and protein antigen responses are present within 2 years of BMT, polysaccharide antibody responses appear last . Streptococcus pneumoniae infection causes significant morbidity and mortality post-BMT . Previous studies have shown good protein antigen responses post-BMT for SCID and PID, but had not examined the polysaccharide responses . We retrospectively analysed pneumococcal polysaccharide (PPS) responses in our patient series.In total, 22 SCID and 12 non-SCID PID were evaluated, all >2 years post BMT: 17 SCID, 12 PID received chemotherapy conditioning; 17 SCID, three PID had T-cell depleted (TCD) BMT, others had nonconditioned whole marrow BMT . All had normal Haemophilus influenza B and tetanus antibody responses . Of 22 SCID, 13 vs 11/12 PID responded to PPS vaccine (P=0.05) . There was no association with donor age, GvHD, B-cell chimerism, or IgG2 level . Fewer TCD marrow recipients responded to PPS (P=0.04) . Analysis of the SCID group showed no association of PPS response with type of marrow received . This is the first study to specifically examine PPS antibody responses following SCID and PID BMT . Pneumococcal conjugate vaccine antibody responses should be examined in these children.

Pediatrics, 2003 Jul, 112(1 Pt 1), e39 - 45
The immunization delivery effectiveness assessment score: a better immunization measure?
Glauber JH.
BACKGROUND: Childhood immunization measures, such as the Health Employer Data Information Set (HEDIS) or the National Immunization Survey, assess the percentage of children up-to-date for a specified series of vaccinations . In particular, the HEDIS assesses immunization delivery to children enrolled in managed care organizations (MCO) . Such measures do not assess the timeliness of immunization delivery with reference to recommended age standards . To achieve maximal protection against vaccine-preventable diseases, children should receive all immunizations within recommended age intervals-fully "on-time." OBJECTIVE: The Immunization Delivery Effectiveness Assessment (IDEA) is a novel immunization measure that assesses, on a continuous scale, the timeliness of administration of each vaccination with reference to recommended age intervals . Specifically we ask: 1) Do absolute immunization rates differ between HEDIS and IDEA? 2) Does relative MCO performance differ when assessed by the 2 performance measures? 3) How well do MCOs perform relative to the standard of fully on-time immunization? The health services implications of using the timeliness standard to assess childhood immunization delivery is discussed . METHODS: A vaccine-dose IDEA score was developed for each of the 14 vaccination events in the 4:3:1:3:3 combination series (4 diphtheria-pertussis-tetanus:3 polio:1 measles-mumps-rubella:3 Haemophilus influenzae type B:3 hepatitis B) . Assessing the actual age of administration with reference to the recommended age of administration generates the vaccine-dose IDEA score . A child's composite IDEA score is obtained by averaging the 14 vaccine-dose IDEA scores . These composite IDEA scores, when averaged among children sampled within the MCO, constitute the MCO's immunization score . SETTING: Retrospective analysis of childhood immunization datasets from a convenience sample of 6 MCOs in 5 states . RESULTS: HEDIS rates ranged from 57% to 75% . IDEA scores ranged from 80% to 90% . Relative rankings of MCO immunization performance were different using HEDIS rates and IDEA scores, respectively . At most, 16% of children in any of these MCOs received all of their immunizations fully on-time . From 47% to 77% of children experienced at least 3 delayed immunizations . CONCLUSIONS: An immunization measure based on timeliness of administration yields both absolute and relative differences in MCO childhood immunization performance when compared with HEDIS rates . By assessing delivery of each component vaccination, the IDEA score permits more detailed analysis of immunization patterns within an MCO and focuses improvement efforts.

Pediatrics, 2003 Jul, 112(1 Pt 1), 193 - 8
Immunization of preterm and low birth weight infants . American Academy of Pediatrics Committee on Infectious Diseases; Saari TN; American Academy of Pediatrics Committee on Infectious Diseases; Preterm (PT) infants are at increased risk of experiencing complications of vaccine-preventable diseases but are less likely to receive immunizations on time . Medically stable PT and low birth weight (LBW) infants should receive full doses of diphtheria, tetanus, acellular pertussis, Haemophilus influenzae type b, hepatitis B, poliovirus, and pneumococcal conjugate vaccines at a chronologic age consistent with the schedule recommended for full-term infants . Infants with birth weight less than 2000 g may require modification of the timing of hepatitis B immunoprophylaxis depending on maternal hepatitis B surface antigen status . All PT and LBW infants benefit from receiving influenza vaccine beginning at 6 months of age before the beginning of and during the influenza season . All vaccines routinely recommended during infancy are safe for use in PT and LBW infants . The occurrence of mild vaccine-attributable adverse events are similar in both full-term and PT vaccine recipients . Although the immunogenicity of some childhood vaccines may be decreased in the smallest PT infants, antibody concentrations achieved usually are protective.

J Bacteriol, 2003 Jul, 185(14), 4152 - 62
Crystal structures of active fully assembled substrate- and product-bound complexes of UDP-N-acetylmuramic acid:L-alanine ligase (MurC) from Haemophilus influenzae; Mol CD et al.; UDP-N-acetylmuramic acid:L-alanine ligase (MurC) catalyzes the addition of the first amino acid to the cytoplasmic precursor of the bacterial cell wall peptidoglycan . The crystal structures of Haemophilus influenzae MurC in complex with its substrate UDP-N-acetylmuramic acid (UNAM) and Mg(2+) and of a fully assembled MurC complex with its product UDP-N-acetylmuramoyl-L-alanine (UMA), the nonhydrolyzable ATP analogue AMPPNP, and Mn(2+) have been determined to 1.85- and 1.7-A resolution, respectively . These structures reveal a conserved, three-domain architecture with the binding sites for UNAM and ATP formed at the domain interfaces: the N-terminal domain binds the UDP portion of UNAM, and the central and C-terminal domains form the ATP-binding site, while the C-terminal domain also positions the alanine . An active enzyme structure is thus assembled at the common domain interfaces when all three substrates are bound . The MurC active site clearly shows that the gamma-phosphate of AMPPNP is positioned between two bound metal ions, one of which also binds the reactive UNAM carboxylate, and that the alanine is oriented by interactions with the positively charged side chains of two MurC arginine residues and the negatively charged alanine carboxyl group . These results indicate that significant diversity exists in binding of the UDP moiety of the substrate by MurC and the subsequent ligases in the bacterial cell wall biosynthesis pathway and that alterations in the domain packing and tertiary structure allow the Mur ligases to bind sequentially larger UNAM peptide substrates.

J Bacteriol, 2003 Jul, 185(14), 4144 - 51
The crystal structure of shikimate dehydrogenase (AroE) reveals a unique NADPH binding mode; Ye S et al.; Shikimate dehydrogenase catalyzes the NADPH-dependent reversible reduction of 3-dehydroshikimate to shikimate . We report the first X-ray structure of shikimate dehydrogenase from Haemophilus influenzae to 2.4-A resolution and its complex with NADPH to 1.95-A resolution . The molecule contains two domains, a catalytic domain with a novel open twisted alpha/beta motif and an NADPH binding domain with a typical Rossmann fold . The enzyme contains a unique glycine-rich P-loop with a conserved sequence motif, GAGGXX, that results in NADPH adopting a nonstandard binding mode with the nicotinamide and ribose moieties disordered in the binary complex . A deep pocket with a narrow entrance between the two domains, containing strictly conserved residues primarily contributed by the catalytic domain, is identified as a potential 3-dehydroshikimate binding pocket . The flexibility of the nicotinamide mononucleotide portion of NADPH may be necessary for the substrate 3-dehydroshikimate to enter the pocket and for the release of the product shikimate.

J Bacteriol, 2003 Jul, 185(14), 4031 - 7
Crystal structure of the YchF protein reveals binding sites for GTP and nucleic acid; Teplyakov A et al.; The bacterial protein encoded by the gene ychF is 1 of 11 universally conserved GTPases and the only one whose function is unknown . The crystal structure determination of YchF was sought to help with the functional assignment of the protein . The YchF protein from Haemophilus influenzae was cloned and expressed, and the crystal structure was determined at 2.4 A resolution . The polypeptide chain is folded into three domains . The N-terminal domain has a mononucleotide binding fold typical for the P-loop NTPases . An 80-residue domain next to it has a pronounced alpha-helical coiled coil . The C-terminal domain features a six-stranded half-barrel that curves around an alpha-helix . The crablike three-domain structure of YchF suggests the binding site for a double-stranded nucleic acid in the cleft between the domains . The structure of the putative GTP-binding site is consistent with the postulated guanine specificity of the protein . Fluorescence measurements have demonstrated the ability of YchF to bind a double-stranded nucleic acid and GTP . Taken together with other experimental data and genomic analysis, these results suggest that YchF may be part of a nucleoprotein complex and may function as a GTP-dependent translation factor.

FEMS Immunol Med Microbiol, 2003 Jul 15, 37(2-3), 185 - 92
Construction of recombinant S-layer proteins (rSbsA) and their expression in bacterial ghosts--a delivery system for the nontypeable Haemophilus influenzae antigen Omp26; Riedmann EM et al.; This study has investigated the feasibility of a combination of recombinant surface layer (S-layer) proteins and empty bacterial cell envelopes (ghosts) to deliver candidate antigens for a vaccine against nontypeable Haemophilus influenzae (NTHi) infections . The S-layer gene sbsA from Bacillus stearothermophilus PV72 was used for the construction of fusion proteins . Fusion of maltose binding protein (MBP) to the N-terminus of SbsA allowed expression of the S-layer in the periplasm of Escherichia coli . The outer membrane protein (Omp) 26 of NTHi was inserted into the N-terminal and C-terminal regions of SbsA . The presence of the fused antigen Omp26 was demonstrated by Western blot experiments using anti-Omp26 antisera . Electron microscopy showed that the recombinant SbsA maintained the ability to self-assemble into sheet-like and cylindrical structures . Recombinant E . coli cell envelopes (ghosts) were produced by the expression of SbsA/Omp26 fusion proteins prior to gene E-mediated lysis . Intraperitoneal immunization with these recombinant bacterial ghosts induced an Omp26-specific antibody response in BALB/c mice . These results demonstrate that the NTHi antigen, Omp26, was expressed in the S-layer self-assembly product and this construct was immunogenic for Omp26 when administered to mice in bacterial cell envelopes.

J Endotoxin Res, 2003, 9(3), 131 - 44
The lipo-oligosaccharides of Haemophilus influenzae: an interesting array of characters; Swords WE et al.; The composition of the lipo-oligosaccharide (LOS) of Haemophilus influenzae is highly variable, especially in the oligosaccharide region . Many of the biosynthetic and transferase genes involved in LOS biosynthesis vary in seemingly random fashion by means of polymerase stuttering within redundant sequences in the 5'-portion of the genes . This results in a heterogeneous population of individual bacteria expressing literally thousands of LOS glycoforms . The simultaneous variation in the expression and structural context of a large number of individual carbohydrate and lipid structures within the LOS yields a diverse array of LOS glycoforms . The expression of glycoforms that mimic host structures may allow the organism to evade innate defenses and to manipulate host cell biology . We review how this randomly generated bacterial combinatorial chemistry results in the production of a large number of carbohydrate structures, in essentially any conceivable structural context, some of which allow the organism to utilize host cell receptors . By generating a diverse population of bacteria expressing different LOS glycoforms, discrete H . influenzae subpopulations may be adapted for survival of different environmental stresses within the airways . Thus, H . influenzae utilizes a simple and efficient "Monte Carlo" strategy for achieving maximal variation in cell surface structures, which allow the organism to adapt efficiently to environmental stresses with a small genome.

Pediatr Int, 2003 Jun, 45(3), 314 - 8
Effect of aluminum adjuvants on safety and immunogenicity of Haemophilus influenzae type b-CRM197 conjugate vaccine; Kanra G et al.; OBJECTIVE: The present study was carried out to evaluate the safety and immunogenicity of the Haemophilus influenzae type b-CRM197 (Hib-CRM197) conjugate vaccine in relation to the change of adjuvant from aluminum hydroxide to aluminum phosphate (AlPO4) . METHODS: The present study was a clinical phase II, observer-blind, randomized, multicenter, controlled study . Subjects were healthy infants aged 6-12 weeks, eligible for expanded program of immunization (EPI) routine vaccination and admitted to Hacettepe University Department of Social Pediatrics and Gulveren Health Center, Ankara . A total of 520 healthy infants were randomized in a 2:2:1 ratio to receive at either Chiron Hib/AlPO4 vaccine or VaxemHib (aluminum hydroxide adjuvant) vaccine or HibTiter (no adjuvant) . Vaccines were administered simultaneously with routine diphtheria, tetanus and pertussis (DTaP) and oral polio vaccine (OPV) vaccines at 2, 4 and 6 months of age . Blood samples for anti-plain polysaccharide (PRP) antibody measurement were collected before the first vaccination and 1 month after the last vaccination . After each vaccination parents filled out a diary for 7 days . RESULTS: Out of 520 subjects enrolled, 514 received three doses and were included for safety analysis . Local and systemic reactions occurred with low and similar frequencies in all groups . Only erythema was more common in Chiron Hib/AlPO4 vaccine (19, 10, 11% in Chiron Hib/AlPO4, VaxemHib and HibTiter, respectively, P < 0.05) . Nine serious adverse events were reported in seven cases of which none were related to vaccines . A total of 504 subjects were included in the immunogenicity analysis . The three vaccines were highly immunogenic and equivalent in terms of percentage of acquisition of long-term protective levels . The anti-PRP geometric mean titers were 9.9, 8.3 and 5.14 micro g/mL, respectively (P < 0.05) . CONCLUSIONS: The use of aluminum compounds adjuvants in Hib-CRM197 conjugate vaccines does not impact the safety profile, while it does increase the magnitude of anti-PRP antibody titers.

Lancet, 2003 Jun 21, 361(9375), 2139 - 48
Bacterial meningitis in children; Saez-Llorens X et al.; This review comprises aspects of the epidemiology, microbiology, pathophysiology, clinical manifestations, diagnosis, management, prognosis, and prevention of bacterial meningitis, with emphasis on the paediatric population . The beginning of this millennium has witnessed the virtual disappearance of Haemophilus invasive disease in some countries, emergence of pneumococcal strains that are resistant to multiple antibiotics, isolation of pneumococci with tolerance to vancomycin, outbreaks and clusters of meningococcal meningitis in several geographical areas, and intense research in development of effective conjugate pneumococcal and meningococcal vaccines . Bacterial meningitis has become an uncommon disease in the developed world . Unfortunately, because of limited economic resources and poor living conditions, many developing countries are still affected by the devastating consequences of this life-threatening systemic infection . Basic and clinical research is needed to discover new antimicrobial and anti-inflammatory agents to improve outcome from disease . Novel strategies are needed to distribute and implement effective vaccines worldwide to prevent bacterial meningitis.

J Infect Dis, 2003 Jul 1, 188(1), 114 - 7 Epub 2003 Jun 23.
Horizontal transfer of the gene encoding outer membrane protein P2 of nontypeable Haemophilus influenzae, in a patient with chronic obstructive pulmonary disease; Hiltke TJ et al.; An adult with chronic obstructive pulmonary disease was monitored prospectively for 2 years . Nontypeable Haemophilus influenzae was isolated from sputum cultures at 22 of 23 monthly clinic visits . Analysis of the isolates, by pulsed-field gel electrophoresis (PFGE), revealed that the patient was colonized by 3 different strains during the 2-year period . The gene encoding outer-membrane protein (OMP) P2, ompP2, was amplified from sputum samples and selected strains obtained from this patient . Analysis of the ompP2 sequences, in combination with the PFGE patterns, indicated that ompP2 horizontal transfer between 2 strains occurred in the respiratory tract, between clinic visits 13 and 14 . Observation of ompP2 horizontal transfer in the human respiratory tract has important implications for both the understanding of ompP2 diversity among strains and the future design of OMP P2-based vaccines.

J Infect Dis, 2003 Jul 1, 188(1), 74 - 80 Epub 2003 Jun 23.
Implications of Haemophilus influenzae biogroup aegyptius hemagglutinins in the pathogenesis of Brazilian purpuric fever; Barbosa SF et al.; Brazilian purpuric fever (BPF) is an acute disease caused by Haemophilus influenzae biogroup aegyptius; it is characterized by fever, purpura, and hypotensive shock and is usually fatal . The factors responsible for bacterial virulence and pathogenesis are poorly known . Hemagglutinins have been frequently associated with bacterial virulence, and, in the present study, hemagglutinating activity was detected in extracellular products from H . influenzae biogroup aegyptius strains isolated from patients with BPF . A 60-kilodalton (kDa) molecule absorbable by human O-type erythrocytes was identified by an immunoblot assay; a corresponding fraction was chromatographically purified, and its pathogenic effect was evaluated . Rabbits injected intravenously with either the whole bacterial extracellular product or the 60-kDa fraction showed reactions similar to those seen in patients with BPF: purpura, congestion, and fibrin thrombi in the inner organs . We suggest that this hemagglutinating factor is one of the major pathogenic components of BPF.

Mol Microbiol, 2003 Jul, 49(1), 241 - 9
Species specificity in the activation of Xer recombination at dif by FtsK; Yates J et al.; In Escherichia coli, chromosome dimers are resolved to monomers by the addition of a single cross-over at a specific locus on the chromosome, dif . Recombination is performed by two tyrosine recombinases, XerC and XerD, and requires the action of an additional protein, FtsK . We show that Haemophilus influenzae FtsK activates recombination by H . influenzae XerCD at H . influenzae dif . However, it cannot activate recombination by E . coli XerCD . Reciprocally, E . coli FtsK cannot activate recombination by the H . influenzae recombinases at H . influenzae dif . We took advantage of this species specificity to gain further insight into the mechanism of activation of Xer recombination at dif by FtsK . We mapped the region of FtsK implicated in species specificity to the extreme 140-amino-acid C-terminal residues of the protein . Our results suggest that FtsK interacts directly with XerCD in order to activate recombination at dif.

Oral Microbiol Immunol, 2003 Aug, 18(4), 256 - 9
Characterization and expression of adjacent proline iminopeptidase and aspartase genes from Eikenella corrodens; Selby T et al.; Two adjacent genes involved in nitrogen metabolism from Eikenella corrodens, with a potential role in pathogenesis, were studied . Proline iminopeptidase (Pip) activity, which may be essential for energy production and protection against host immune mechanisms, is exhibited by E . corrodens . Analysis of Pip-expressing clones revealed an ORF of 939 bases with a predicted amino acid sequence identity of 67% to the Pip of Neisseria gonorrhoea . 200 bp downstream from pip, an ORF of 1395 bases, encoding a protein with 87% identity to a putative aspartase from the Neisseria meningitidis genome sequence, was identified . Enzymatic function was confirmed with a complemented Escherichia coli aspartase deficient mutant . The E . corrodens aspartase was found to be 77% identical to the Haemophilus influenzae aspartase sequence, which was originally identified on the basis of its ability to bind plasminogen . However, the E . corrodens aspartase had no such activity . Southern hybridization indicated both genes to be single copy and conserved within the genomes of a diverse panel of E . corrodens isolates from health and disease.

J Laryngol Otol, 2003 Jun, 117(6), 449 - 53
Increased antimicrobial resistance in organisms recovered from otitis media with effusion; Brook I et al.; Previous studies concerning the microbiology of otitis media with effusion (OME) did not correlate the past use of antimicrobial agents with the recovered organism's antimicrobial susceptibility . A retrospective analysis of cultures obtained from aspirates of 129 children with OME was performed . The study identified the isolated organisms and determined their susceptibility to the most recently administered antimicrobials . Bacterial growth was noted in 58 (45 per cent) patients . Aerobic organisms only were recovered in 37 aspirates (63 per cent of the culture-positive aspirates); anaerobic bacteria in seven (12 per cent); and mixed aerobic and anaerobic bacteria in 14 (24 per cent) . A total of 92 bacterial isolates were recovered, accounting for 1.6 isolates per specimen (1.1 aerobes and 0.5 anaerobes) . There were a total of 66 aerobic isolates, including Haemophilus influenzae non type-b (20 isolates), Streptococcus pneumoniae (17), and Staphylococcus spp . (seven) . Twenty-six anaerobes were recovered, including Peptostreptococcus spp . and Prevotella spp . (eight each) and Propionibacterium acnes (four) . Resistance to the antimicrobial used was found in 60 (65 per cent) isolates, recovered from 41 (71 per cent) of the patients . Of the 41 patients in whom resistance was detected, 37 (90 per cent) had been treated within three months of culture and four (10 per cent) had completed treatment more than three months before the cultures were taken (p < 0.01) . The highest rate of recovery of resistant organisms was following trimethoprim-sulfamethoxazole (96 per cent), amoxycillin (71 per cent), and azithromycin (56 per cent) . Of the patients treated with amoxycillin, H influenzae predominated . S pneumoniae was recovered from four of the seven (57 per cent) after trimethoprim-sulfamethoxazole, four of 14 (29 per cent) following amoxycillin, and three of 11 (27 per cent) after azithromycin . The data illustrate the relationship between resistance to the antimicrobials given to children and their recovery from the middle ear of patients with OME.

Rev Prat, 2003 May 1, 53(9), 985 - 8
{Acute laryngeal dyspnea}; Cros AM et al.; Laryngeal dyspnea is a life-threatening emergency situation . The diagnosis is clinical and made from the association of: inspiratory bradypnea, intercostal and sus-sternal inspiratory depression, with or without stridor . The aetiologies are most often laryngeal tumours or inflammatory oedema; incidence of epiglottitis has decreased due to vaccine against Haemophilus influenzae . Airway obstruction due to foreign body includes acute laryngeal dyspnea and reflex paroxysmal coughing without fever . Management of a laryngeal dyspnea depends on the aetiology and the severity of clinical symptoms . Medical treatment associates racemic epinephrine aerosol, steroids, and oxygenation . In the presence of severe dyspnea, intubation after anaesthetising the patient and positive pressure ventilation is required.

Respir Med, 2003 Jun, 97(6), 625 - 33
Telithromycin in the treatment of community-acquired pneumonia: a pooled analysis; Hagberg L et al.; The efficacy of telithromycin has been assessed in six Phase III studies involving adults with mild to moderate community-acquired pneumonia (CAP) with a degree of severity compatible with oral therapy . Patients received telithromycin 800 mg once daily for 7-10 days in three open-label studies (n=870) and three randomized, double-blind, comparator-controlled studies (n=503) . Comparator antibacterials were amoxicillin 1000 mg three-times daily, clarithromycin 500 mg twice daily and trovafloxacin 200 mg once daily . Clinical and bacteriological outcomes were assessed 7-14 days post-therapy . Among telithromycin-treated patients, per-protocol clinical cure rates were 93.1 and 91.0% for the open-label and comparative studies, respectively . Telithromycin treatment was as effective as the comparator agents . High eradication and clinical cure rates were observed for infections caused by key pathogens: Streptococcus pneumoniae including isolates resistant to penicillin G and/or erythromycin A (95.4%), Haemophilus influenzae (89.5%) and Moraxella catarrhalis (90%) . Telithromycin was also highly effective in patients with infections caused by atypical and/or intracellular pathogens and those at increased risk of morbidity . Telithromycin was generally well tolerated . Telithromycin 800 mg once daily for 7-10 days offers a convenient and well-tolerated first-line oral therapy for the empirical treatment of mild to moderate CAP.

Health Bull (Edinb), 2000 Sep, 58(5), 390 - 5
An audit of the vaccination and antibiotic prophylaxis practices amongst patients splenectomised in Lothian; Pickering J et al.; OBJECTIVE: To evaluate current post-splenectomy immunisation and antibiotic prophylaxis practices amongst patients splenectomised in Lothian between 1st January 1989 and 31st December 1997 . DESIGN: Retrospective cross-sectional study of the general practitioners of patients recorded as having had a splenectomy operation . SETTING: Lothian Health Board (population 771,000) . SUBJECTS: Asplenic patients recorded on Scottish Morbidity Records as having had a splenectomy in Lothian between 1st January 1989 and 31st December 1997 . RESULTS: Eighty point six percent of the patients were vaccinated against Streptococcus pneumoniae, 65.9% were vaccinated against Haemophilus influenzae, and 48.2% were vaccinated against Neisseria meningitidis; 74.8% of patients were being prescribed long-term antibiotic prophylaxis . However, only 37.4% were both vaccinated and prescribed prophylaxis as recommended, in Department of Health guidelines . CONCLUSIONS: The vaccination levels seen in this survey are higher than levels reported in previously published surveys in the UK . Reported rates of post splenectomy vaccination against S . pneumoniae in both the UK and North America in 1993 ranged from 10% to 36% . However, there still remains a substantial proportion of splenectomised patients in Lothian, including some of those with the highest risk of infection, who are not vaccinated or prescribed prophylaxis as recommended.

Diagn Microbiol Infect Dis, 2003 Jun, 46(2), 147 - 50
Re-evaluation of quality control guidelines for gatifloxacin and garenoxacin (BMS284756) when susceptibility testing Haemophilus influenzae and Streptococcus pneumoniae; Anderegg TR et al.; An eight laboratory M23-A2 quality control (QC) study was performed for the re-evaluation of gatifloxacin, a new fluoroquinolone, using disk diffusion tests against Streptococcus pneumoniae ATCC 49619 . The study also re-evaluated garenoxacin, a novel investigational des-F(6)-quinolone, using disk diffusion tests against S . pneumoniae ATCC 49619 and broth MIC for Hemophilus influenzae ATCC 49247 . The gatifloxacin zone diameter results for S . pneumoniae did not indicate a need for QC range modification (26-34 mm; 98.3% of results); however, the garenoxacin zone diameters did demonstrate a need for a minor modification (1 mm; 26-34 mm; 96.3% of reported results) . The broth MIC results for H . influenzae showed that 83.1% of the results were at the upper limit of the current range (0.008 microg/ml) published by the National Committee for Clinical Laboratory Standards (NCCLS) . The proposed correction could either be 0.002-0.015 microg/mlor 0.004-0.015 microg/ml, each encompassing 100.0% of reported results (prior and current studies) . All MIC results for control drugs, levofloxacin and moxifloxacin (disks) or gatifloxacin and clarithromycin (MIC), were within published NCCLS ranges.

Braz J Infect Dis, 2003 Feb, 7(1), 44 - 61
Antibacterial resistance of community-acquired respiratory tract pathogens recovered from patients in Latin America: results from the PROTEKT surveillance study (1999-2000); Mendes C et al.; PROTEKT (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin) is a global surveillance study established in 1999 to monitor antibacterial resistance of respiratory tract organisms . Thirteen centers from Argentina, Brazil and Mexico participated during 1999-2000; they collected 1806 isolates (Streptococcus pneumoniae 518, Haemophilus influenzae 520, Moraxella catarrhalis 140, Staphylococcus aureus 351, S . pyogenes 277) . Overall, 218 (42.1%) of the S . pneumoniae isolates had reduced susceptibility to penicillin, 79 (15.3%) were penicillin-resistant and 79 (15.3%) were erythromycin-resistant . Mexico had the highest prevalence of penicillin (76.5%) and erythromycin (31.2%) resistance . Of 77 erythromycin-resistant S . pneumoniae tested for resistance genotype, 43 possessed mef(A), 33 possessed erm(B) and 1 possessed both erm(B) and mef(A) mechanism . All S . pneumoniae isolates were fully susceptible to telithromycin, linezolid, teicoplanin and vancomycin . Among H . influenzae isolates, 88 (16.9%) produced beta-lactamase, ranging from 11% (Brazil) to 24.5% (Mexico) . Among M . catarrhalis isolates, 138 (98.6%) produced beta-lactamase . Twenty-four (8.7%) of the S . pyogenes isolates were erythromycin-resistant; resistance being attributable to mefA (n=18), ermTR (n=5) and ermB (n=1) . All H . influenzae, M . catarrhalis and S . pyogenes were fully susceptible to telithromycin . Methicillin resistance was found in 26.5% of the S . aureus isolates (Argentina 15%; Mexico 20%; Brazil 31.3%) . Telithromycin was effective against 97.7% of methicillin-susceptible isolates . PROTEKT confirms that antibacterial resistance is an emerging problem in Latin America . The previously reported high levels of pneumococcal resistance to the beta-lactam and macrolides were exceeded . New agents that do not induce resistance or that exert low selective pressure, e.g . telithromycin, are essential to safeguard future antibacterial efficacy.

Commun Dis Intell, 2003, 27 Suppl, S61 - 6
SENTRY Antimicrobial Surveillance Program Asia-Pacific region and South Africa; Bell J et al.; The SENTRY Antimicrobial Surveillance Program was initiated in January 1997 and was designed to monitor the predominant pathogens and antimicrobial resistance for both nosocomial and community-acquired infections globally by using validated, reference-quality identification and susceptibility testing methods performed in a central laboratory . Consecutive bacterial or fungal isolates, deemed clinically significant by local criteria, are forwarded to the local reference laboratory from various study objectives . The major objectives include blood stream infections, community-acquired respiratory tract infections (Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis), pneumonias in hospitalised patients, skin and soft tissue infections, and urinary tract isolates from hospitalised patients . In 2001, special objectives were introduced to examine gastroenteritis pathogens and beta-haemolytic streptococcal isolates . Over 22 nations participate in SENTRY surveillance globally . The Women's and Children's Hospital, Adelaide has been the reference centre for the Asia-Pacific region and South Africa since 1998, and three other Australian institutions, from Brisbane, Perth, and Adelaide, are part of the global network . All isolates received from our region are tested against up to 29 antimicrobial agents using custom-made broth microdilution panels . The data generated from SENTRY allows Australia to compare our antimicrobial resistance patterns and trends with our regional neighbours.

Otol Neurotol, 2003 May, 24(3), 353 - 7
Detection of Chlamydia pneumoniae in cholesteatoma tissue: any pathogenetic role?
Ronchetti F, Ronchetti R, Guglielmi F, Chiappini I, Contini C, Filipo R, Santino I, Cerruto R, Bernardeschi D, Barbara M.
BACKGROUND: Acquired cholesteatoma is a complication of chronic otitis media that is usually associated with an intense local inflammatory reaction . Cholesteatoma probably arises from epithelial migration close to an ongoing host inflammatory response attributable to a chronic bacterial infection . Chlamydia pneumoniae is an intracellular microorganism associated with several pathologic conditions originally considered noninflammatory, including asthma, atherosclerosis, and Alzheimer disease . To investigate a possible relationship between C . pneumoniae and the development of cholesteatoma, tissue was studied in three different layers by polymerase chain reaction analysis . The results were compared with those relative to other two common middle-ear pathogens, Mycoplasma pneumoniae and Haemophilus influenzae . METHODS: Cholesteatoma specimens were collected from 32 patients undergoing middle ear surgery . A series of 5 microm-thick specimens were obtained at three different tissue levels, internal (matrix), intermediate (perimatrix), and external (granulation tissue), and processed by polymerase chain reaction for detection of C . pneumoniae, H . influenzae, and M . pneumoniae . Fragmentation and polymerase chain reaction amplification were carried out using two substantially different techniques.RESULTS: C . pneumoniae was detected with either polymerase chain reaction techniques in the internal layers in 16 of the 32 cholesteatomas (50%), associated with a positive finding in the intermediate layer in two cases and in the external layer in one case . Four specimens contained H . influenzae, always in the external layer, whereas none contained M . pneumoniae . CONCLUSIONS: The close relationship between cholesteatoma and C . pneumoniae demonstrated by the findings of this study could suggest a direct cause and effect link between the pathogen action and the clinical manifestations . Otherwise, a facilitated colonization by C . pneumoniae and chronic pathology of the ear could both take origin from a peculiar immunologic background of the host.

Vaccine, 2003 Jul 4, 21(23), 3330 - 4
Comparative reactogenicity and parental acceptability of pertussis vaccines administered into the ventrogluteal area and anterolateral thigh in children aged 2, 4, 6 and 18 months; Cook IF et al.; The importance of site of injection of combined pertussis/diphtheria/tetanus vaccines was investigated in two single blind studies.In the pilot study, in which the research instrument was trialed, 283 children aged 2-18 months received whole cell pertussis vaccine (DTPw) by the intramuscular route either into the anterolateral thigh or the ventrogluteal site . In the larger randomised study, 566 children aged 2-18 months were similarly injected with acellular pertussis vaccine (DTPa) . Adverse reactions monitored after 24h showed the same lower rates for both vaccines with ventrogluteal injection compared with anterolateral thigh injection for systemic reactions (irritability (P<0.0001), perceived fever (P<0.0001), persistent crying/screaming (P<0.0001) and local reactions (bruising (P<0.0001) and redness/swelling (P<0.0001)) . The Haemophilus influenzae type b vaccine (HibTITER) given concurrently in the contralateral site to the pertussis vaccine showed the same lower rates in both studies for ventrogluteal injection compared with anterolateral thigh injection for local reactions (redness/swelling both studies (P<0.0001) and bruising DTPw study (P<0.0001) and DTPa study (P<0.0004)).Parental acceptability was greater (P<0.0001) in both studies for ventrogluteal injection compared with anterolateral thigh injection.

Eur J Epidemiol, 2003, 18(4), 363 - 7
Invasive Haemophilus influenzae disease: an ecological study of sociodemographic risk factors before and after the introduction of Hib conjugate vaccine; Olowokure B et al.; This study examines the impact of H . influenzae type b (Hib) conjugate vaccine on sociodemographic risk factors for invasive H . influenzae disease in the 2 years before and immediately after the introduction of Hib conjugate vaccine . An ecological study design was used and cases were identified using active surveillance employing several surveillance systems . The study population comprised all children aged < 5 years resident in the West Midlands, an English health region, with laboratory confirmed invasive disease 2 years before (1990-1992) and 2 years after (1992-1994) the introduction of Hib conjugate vaccine . Selected sociodemographic variables derived from the UK census were obtained for all census enumeration districts in the region . Each variable was then ranked and divided into six categories . Linear associations between disease rates and sociodemographic variables were examined . Overall, there was a significant reduction in the incidence of invasive H . influenzae disease . In the pre-conjugate vaccine era there were trends of decreasing disease incidence with increasing child population density (p = 0.012) and total population density (p = 0.0023) . In the post-conjugate vaccine period, total population density (p = 0.0275) remained significant and a trend of increasing disease incidence with increasing population mobility (p = 0.0012) was seen . Although Hib conjugate vaccine has resulted in a dramatic reduction in disease incidence changes in sociodemographic risk factors were identified in the post-conjugate vaccine period, particularly population mobility . Our results may have implications for current and future vaccine strategies.

Stat Med, 2003 Jun 30, 22(12), 1989 - 98
Implications of genetic traits on vaccine efficacy; Murthy BN; Literature on genetic screening in the community suggests that people having specific genotypes may either get or protect from infection, for example, malaria, human papilloma virus, and haemophilic influenza, for which vaccines are either already developed or being targeted . In such a situation, the evaluation of the efficacy of vaccine in the community needs to be examined with caution . In this paper, I present a method for the estimation of vaccine efficacy (VE) in the presence of genetic traits/component (theta) and the sample size required to estimate the 95 per cent CI with a given relative width for the estimated vaccine efficacy . Considering true efficacy ranging from 40 to 80 per cent and the possible values of the genetic component (theta) ranging from 0 to 60 per cent, the VE was estimated . The 95 per cent confidence intervals (CI) for the estimated VE for relative widths (R) 1.0 and 0.1 were computed . The sample sizes required for each of the unvaccinated and vaccinated cohorts were computed for estimating the 95 per cent CI for given incidence rates in the unvaccinated (Iu) cohort . In the presence of genetic traits I found that the VE was consistently overestimated . There existed change in the location as well as the asymmetry of the 95 per cent CIs over the point estimate of VE . The sample size required for estimating 95 per cent CI of VE was substantially reduced, resulting in savings . The more the genetic component (theta) affecting disease in the community, the more the savings in sample size . I examined the above estimators for (i) VE, (ii) 95 per cent CI for VE and (iii) sample size required for estimating 95 per cent CI of VE using the real-life data from the Haemophilus influenzae type b vaccine trial conducted in Finland and the global genetic structure of encapsulated H . influenza . Because of escalated VE and large savings in sample size for estimating the 95 per cent CI for VE, I recommend that the design should consider the genetic component that causes/protects from infection/disease for the evaluation of efficacy of vaccine in the field .

Mol Gen Mikrobiol Virusol, 2003, (2), 21 - 5
{Multilocus sequence-typing for characterization of Moscow strains of Haemophilus influenzae type b}; Platonov AE et al.; Haemophilius influenzae, type b (Hib) bacteria, were genotyped by multilocus sequence typing (MLST) using 5 loci (adk, fucK, mdh, pgi, recA) . 42 Moscow Hib strains (including 38 isolates form cerebrospinal fluid of children, who had purulent meningitis in 1999-2001, and 4 strains isolated from healthy carriers of Hib), as well as 2 strains from Yekaterinburg were studied . In MLST a strain is characterized, by alleles and their combinations (an allele profile) referred to also as sequence-type (ST) . 9 Sts were identified within the Russian Hib bacteria: ST-1 was found in 25 strains (57%), ST-12 was found in 8 strains (18%), ST-11 was found in 4 strains (9%) and ST-15 was found in 2 strains (4.5%); all other STs strains (13, 14, 16, 17, 51) were found in isolated cases (2.3%) . A comparison of allelic profiles and of nucleotide sequences showed that 93% of Russian isolates, i.e . strain with ST-1, 11, 12, 13, 15 and 17, belong to one and the same clonal complex . 2 isolates from Norway and Sweden from among 7 foreign Hib strains studied up to now can be described as belonging to the same clonal complex; 5 Hib strains were different from the Russian ones.

Pediatr Infect Dis J, 2003 Jun, 22(6), 557 - 62
HACEK endocarditis in infants and children: two cases and a literature review; Feder HM Jr et al.; We report 2 cases of Haemophilus parainfluenzae endocarditis and review 34 cases of HACEK endocarditis from the literature . HACEK organisms are the most common cause of Gram-negative endocarditis in children . They have a propensity to form friable vegetations (especially H . parainfluenzae) that break off and cause symptomatic emboli . HACEK endocarditis (from a review of the 36 published cases) may involve previously normal hearts (33%), may be complicated by embolization (31%) and may require vegetectomy or other surgery (31%) . Mortality with HACEK endocarditis was 14% . HACEK organisms may be resistant to penicillins but are susceptible to third generation cephalosporins.

Pediatr Infect Dis J, 2003 Jun, 22(6), 509 - 15
Can acute otitis media caused by Haemophilus influenzae be distinguished from that caused by Streptococcus pneumoniae?
Leibovitz E, Satran R, Piglansky L, Raiz S, Press J, Leiberman A, Dagan R.
BACKGROUND: Previous limited data suggest that acute otitis media (AOM) caused by Streptococcus pneumoniae can present as a more severe disease than that caused by Haemophilus influenzae or Moraxella catarrhalis, as expressed by both tympanic membrane and systemic findings . OBJECTIVES: To evaluate the severity of disease and impact of various pathogens, age, disease history and previous antibiotic therapy in children with AOM by using a comprehensive clinical/otologic score . PATIENTS AND METHODS: The study group consisted of 372 children ages 3 to 36 months with AOM seen at the pediatric emergency room during 1996 through 2001 . All patients had tympanocentesis and middle ear fluid culture performed at enrollment . Clinical status was determined by a clinical/otologic score evaluating severity (0 = absent to 3 = severe) of tympanic membrane findings (redness and bulging) and patient's fever, irritability and ear tugging . Maximal severity score was 15 . RESULTS: There were 138 (37%) H . influenzae, 76 (21%) S . pneumoniae, 64 (17%) mixed infections (H . influenzae + S . pneumoniae) and 94 (25%) culture-negative cases . The overall clinical/otologic score was higher in culture-positive than in culture-negative patients (9.27 +/- 2.75 vs.8.38 +/- 3.08, P = 0.01) . When analyzed by age groups, this difference was significant only for the youngest age group (3 to 6 months, P = 0.05) . The severity scores for AOM caused by H . influenzae and S . pneumoniae were significantly higher than in the culture-negative AOM when tympanic membrane redness and bulging were analyzed separately . No differences were recorded in clinical/otologic scores between different pathogens (9.49 +/- 2.86, 9.03 +/- 2.72 and 9.09 +/- 2.54 for H . influenzae, S . pneumoniae and H . influenzae + S . pneumoniae, respectively) . The mean clinical/otologic score was higher in culture-positive than in culture-negative patients without relationship to previous antibiotic treatment or number of previous AOM episodes . CONCLUSIONS: (1) The clinical/otologic score of culture-positive young infants was higher than that of culture-negative infants; (2) the severity of tympanic membrane redness and bulging were the most indicative factors discriminating between a bacterial and nonbacterial etiology of AOM; and (3) the use of a clinical/otologic score could not discriminate among various bacterial etiologies of AOM.

Vaccine, 2003 Jun 20, 21(21-22), 2906 - 10
The immunogenicity of oral poliomyelitis vaccine in a primary vaccination series at 2, 4 and 6 months given concurrently with Hib, hepatitis B and diphtheria, tetanus and whole-cell pertussis vaccines administered as three separate injections or as a combination pentavalent vaccine; Hogg K et al.; The increasing number of infant immunisations has spurred development of novel combination vaccines . This investigation assesses the immunogenicity of oral poliomyelitis vaccine (OPV) under current and possible new conditions, to help ensure vaccination regimes continue to provide optimal protection against polio in the final stages of polio eradication . Neutralising antibody titres were measured in approximately 200 infants immunised with OPV at 2, 4 and 6 months in tandem with either a combined pentavalent liquid Haemophilus influenza B (Hib), hepatitis B, diphtheria, tetanus and whole-cell pertussis vaccine or three separate but concurrently administered licensed vaccines (diphtheria, tetanus and whole-cell pertussis (DTP), lyophilised Hib, and hepatitis B) . Following three doses of OPV, at least 98% of infants demonstrated neutralising antibodies at 1:8 to each poliovirus type under both vaccination regimes, and geometric mean titres (GMTs) well above the suggested protective titre were also observed for all poliovirus types . OPV appears to be effective not only in producing protective antibody titres in an extremely high proportion of infants when given in combination with currently licensed vaccines, but also when administered together with the combination pentavalent vaccine under study . This is encouraging for the continued role of OPV in infant immunisation.

Infect Genet Evol, 2001 Jul, 1(1), 29 - 39
Dynamics of bacterial colonisation in the respiratory tract of patients with cystic fibrosis; Renders N et al.; Mutations in the human genome may result in altered phenotypes . The cystic fibrosis (CF) patient, for instance, suffers from an aberrant composition of the epithelial lining of the gastrointestinal and respiratory tract . In this particular case, a single point mutation in the cystic fibrosis conductance regulator (CFTR) gene results in major physiological changes resulting in ecological changes that generate a niche particularly attractive to a selected set of microbial pathogens . We here present a review on the dynamics of the bacterial populations inhabiting the CF lung . Studies focusing on Staphylococcus aureus, Haemophilus influenzae and Pseudomonas aeruginosa will be summarised and discussed, whereas the technology used for microbial characterisation will be shortly highlighted . Emphasis, however, will be on those studies that assessed the genetic diversity among clinical isolates that were obtained over prolonged periods of time, enabling the distinction between persistent colonisation versus frequent re-infection by the selected pathogens . Evolutionary adaptation of pathogens to the CF lung is a common theme in many of these studies.

Infect Genet Evol, 2003 May, 3(1), 57 - 66
High rates of recombination in otitis media isolates of non-typeable Haemophilus influenzae; Cody AJ et al.; Non-typeable (NT) or capsule-deficient, Haemophilus influenzae (Hi) is a common commensal of the upper respiratory tract of humans and can be pathogenic resulting in diseases such as otitis media, sinusitis and pneumonia . The lipopolysaccharide (LPS) of NTHi is a major virulence factor that displays substantial intra-strain and inter-strain variation of its oligosaccharide structures . To investigate the genetic basis of LPS variation we sequenced internal regions of each of seven genes required for the biosynthesis of either the inner or the outer core oligosaccharide structures . These sequences were obtained from 25 representative NTHi isolates from episodes of otitis media . We found abundant evidence of recombination among LPS genes of NTHi, a finding in marked contrast to previous analyses of biosynthetic genes for capsular polysaccharide, a well-documented virulence factor of Hi . We found mosaic sequences, linkage equilibrium between loci and a lack of congruence between gene trees . These high rates were not confined to LPS genes since evidence for similar amounts of recombination was also found in eight housekeeping genes in a subset of the same 25 isolates . These findings provide a population based foundation for a better understanding of the role of NTHi LPS as a virulence factor and its potential as a candidate vaccine.

Sex Transm Infect, 2003 Jun, 79(3), 202 - 7
Increasing relative prevalence of HSV-2 infection among men with genital ulcers from a mining community in South Africa; Lai W et al.; OBJECTIVES: To determine the aetiology of genital ulcer disease (GUD) and its association with HIV infection in the mining community of Carletonville, South Africa, from two cross sectional surveys of consecutive men presenting with genital lesions during October 1993 to January 1994 and July to November 1998 . METHODS: A multiplex polymerase chain reaction (M-PCR) assay combined with amplicon detection was used to identify DNA specific sequences of Treponema pallidum, herpes simplex virus (HSV), and Haemophilus ducreyi . A real time PCR assay was used to differentiate between HSV-1 and HSV-2 . RESULTS: M-PCR detected T pallidum, HSV, and H ducreyi in 10.3%, 17.2%, and 69.4% of 232 GUD patients during 1993-4 and in 12.4%, 36.0%, and 50.5% of 186 GUD patients in 1998 . The proportion of patients with more than one agent increased significantly from 7.3% (17/232) in 1993-4 to 16.7% (31/186) in 1998 (p <0.01) . HSV-2 was detected in a higher proportion of ulcer specimens from HIV infected patients than in specimens from HIV uninfected patients during both time periods (1993-4: 26.2% v 6.7%, p <0.001; 1998: 42.1% v 29.6%, p >0.09) . CONCLUSIONS: Based on two cross sectional surveys, 4 years apart, chancroid remained the leading cause of GUD in men who presented at the STD clinic with genital ulcers in the mining community of Carletonville, South Africa . The relative prevalence of primary syphilis has remained low . However, HSV-2 has emerged as a more significant cause of GUD and the proportion of GUD patients infected with more than one agent also increased significantly . HSV-2 DNA was detected in a significantly higher proportion of ulcer specimens from HIV positive patients than from HIV negative patients . No association was found between HIV infection status and the relative prevalence of chancroid or syphilis.

Pediatr Infect Dis J, 2003 May, 22(5), 405 - 13
Bacteriologic and clinical efficacy of high dose amoxicillin for therapy of acute otitis media in children; Piglansky L et al.; BACKGROUND: High dose (70 to 90 mg/kg/day) amoxicillin is recommended as first line therapy of acute otitis media (AOM) in geographic areas where drug-resistant Streptococcus pneumoniae is prevalent . Information on the bacteriologic efficacy of high dose amoxicillin treatment for AOM is limited . OBJECTIVES: To evaluate the bacteriologic and clinical efficacy of high dose amoxicillin as first line therapy in AOM . METHODS: In a prospective study 50 culture-positive patients ages 3 to 22 months (median, 9 months; 77% <1 year) were treated with high dose amoxicillin (80 mg/kg/day three times a day for 10 days) No antibiotics were administered 72 h before enrollment . Twenty-four (48%) patients presented with their first episode of AOM . Middle ear fluid was cultured by tympanocentesis at enrollment and on Days 4 to 6 of therapy . Additional middle ear fluid cultures were obtained if clinical relapse occurred . Bacteriologic failure was defined by positive cultures on Days 4 to 6 and clinical failure by no change or worsening of AOM signs and symptoms and requirement for additional antibiotics during therapy and/or at end of therapy . Patients were followed until Day 28 +/- 2 . Susceptibility to penicillin and amoxicillin was measured by E-test . RESULTS: Sixty-five organisms were recovered at enrollment: Haemophilus influenzae (38), Streptococcus pneumoniae (24), Streptococcus pyogenes (2) and Moraxella catarrhalis (1) . Eighteen (75%) S . pneumoniae were nonsusceptible to penicillin (MIC > 0.1 microg/ml) . All 24 S . pneumoniae isolates had amoxicillin MIC < or = 2.0 microg/ml . Thirteen (34%) of the 38 H . influenzae were beta-lactamase producers . Eradication was achieved in 41 (82%) patients for 54 of 65 (83%) pathogens: 22 of 24 (92%) S . pneumoniae, 21 of 25 (84%) beta-lactamase-negative H . influenzae, 8 of 13 (62%) beta-lactamase-positive H . influenzae, 2 of 2 S . pyogenes and 1 of 1 M . catarrhalis . Seven organisms not initially present were isolated on Days 4 to 6 in 5 patients: 3 beta-lactamase-positive H . influenzae; 1 beta-lactamase-negative H . influenzae; 2 S . pneumoniae; and 1 M . catarrhalis . In total 14 of 50 (28%) patients failed bacteriologically on Days 4 to 6 (persistence + new infection), of whom 9 (64%) had beta-lactamase-positive H . influenzae . Three (33%) of the 9 patients with bacteriologic failure (2 beta-lactamase-positive H . influenzae, 1 S . pneumoniae) failed also clinically on Days 4 to 6 . CONCLUSIONS: The predominant pathogens isolated from children with AOM failing high dose amoxicillin therapy were beta-lactamase-producing organisms . Because its overall clinical efficacy is good, high dose amoxicillin is still an appropriate choice as first line empiric therapy for AOM, followed by a beta-lactamase-stable drug in the event of failure.

Int J Antimicrob Agents, 2003 Jun, 21(6), 581 - 4
The in vitro effects of faropenem on lower respiratory tract pathogens isolated in the United Kingdom; Walsh F et al.; Faropenem is a new oral penem with a structure different from current beta-lactams including carbapenems . The susceptibility of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis to faropenem, a macrolide, a beta-lactam, a beta-lactam/beta-lactamase inhibitor combination and two fluoroquinolones was investigated . S . pneumoniae was the most susceptible of the three species to faropenem . The MIC(90)s of faropenem against M . catarrhalis and H . influenzae were 0.5 and 1 mg/l, respectively . They were similar to amoxiclav (MIC (90)s of 0.25 and 0.5 mg/l) . The quinolones showed strong activity against H . influenzae . A cluster analysis of the activities of amoxycillin and faropenem demonstrated a direct relationship between the two antimicrobial agent's activities and resistance profiles against both S . pneumoniae and H . influenzae.

Int J Antimicrob Agents, 2003 Jun, 21(6), 501 - 9
Resistant Haemophilus influenzae in community-acquired respiratory tract infections: a role for cefixime; Verhoef J et al.; An increase in Haemophilus influenzae resistance has been documented around the world during the last 30 years . Resistance is due to the production of beta-lactamases, and/or changes to penicillin-binding protein (PBP) targets . The resistance problem has led to the need for new therapeutic strategies aimed at maintaining effective management of both upper respiratory tract infections (URTIs) and lower respiratory tract infections (LRTIs) . Among antimicrobial agents tested, third-generation cephalosporins have been shown to possess excellent in vitro activity against beta-lactamase-positive and -negative isolates, corresponding with proven clinical efficacy in a wide range of RTIs . The role of H . influenzae in RTIs is outlined, changing trends in epidemiological surveillance studies monitored and implications for therapy, based upon results of clinical trials discussed.

IDrugs, 2003 Mar, 6(3), 240 - 5
The future prospects of oxazolidinones; Johnson AP; The high rates of antimicrobial resistance seen among many Gram-positive pathogens means that there is an ongoing need for new antibacterial drugs . Currently, several pharmaceutical companies are investigating compounds belonging to a new class of anti-Gram-positive agents, the oxazolidinones, one member of which, linezolid, is licensed for clinical use . Interest in oxazolidinones is being stimulated by the results of recent trials demonstrating the excellent clinical efficacy of linezolid in a range of infections . Linezolid is also a relatively safe drug, the main toxicity issue relating to development of thrombocytopenia in some patients receiving prolonged courses . With regard to new oxazolidinones, there is particular interest in analogs that, in early laboratory evaluation, show enhanced activity against the respiratory pathogens Haemophilus influenzae and Moraxella catarrhalis . The future potential of the oxazolidinones will depend on the antibacterial spectra, pharmacokinetic properties and toxicity profiles of new compounds, and the degree to which their clinical use may be affected by the emergence of oxazolidinone-resistant pathogens.

Microb Pathog, 2003 Jun, 34(6), 287 - 96
Functional characterization of HgbB, a new hemoglobin binding protein of Pasteurella multocida; Cox AJ et al.; The biological function and role in pathogenesis of a Pasteurella multocida A:1 strain hemoglobin binding protein was investigated . The hgbB gene from the P . multocida A:1 strain, VP161, was cloned and characterized . hgbB was 2991 bp in length and encoded a mature length protein of 111 kDa . HgbB was predicted to be an outer membrane protein and shared 68 and 69% similarity to the hemoglobin/hemoglobin-haptoglobin binding protein, HI0712 from Haemophilus influenzae Rd and HgpC, from H . influenzae b, respectively . HgbB exhibited features typical of TonB dependent receptors, including seven conserved regions typical of these proteins, and conserved invariant residues . Escherichia coli expressing recombinant HgbB was found to bind hemoglobin in a solid phase dot blot binding assay . However, when a truncated form of the protein was expressed in E . coli, cells could no longer bind hemoglobin . Insertional inactivation of hgbB did not affect the ability of P . multocida to bind hemoglobin, nor its ability to produce disease in a mouse model . In addition, recombinant HgbB did not confer any protection against homologous or heterologous challenge.

Microb Pathog, 2003 Jun, 34(6), 261 - 5
S-carboxymethylcysteine inhibits the attachment of Streptococcus pneumoniae to human pharyngeal epithelial cells; Cakan G et al.; Streptococcus pneumoniae causes respiratory and other invasive infections . Increased resistance of this bacterium to antibiotics necessitates new approaches to the treatment of infections . Attachment of bacteria to human pharyngeal epithelial cells is the initial step in the pathogenesis of infection and S-carboxymethylcysteine (S-CMC) can modulate the attachment of Moraxella catarrhalis and nontypable Haemophilus influenzae to epithelial cells . Unlike these two, S . pneumoniae is gram-positive and has a well-defined capsule . Here we examined the effects of S-CMC on the attachment and detachment of S . pneumoniae to human pharyngeal epithelial cells in vitro . Treatment of these cells with S-CMC significantly reduced the number of attached S . pneumoniae . S-CMC also resulted in a significant increase in the detachment of already attached S . pneumoniae to epithelial cells . In addition, treatment of S . pneumoniae with S-CMC significantly reduced their ability to attach to epithelial cells, but not the number of viable bacteria . Our study shows that S-CMC modulates the attachment of S . pneumoniae to human pharyngeal epithelial cells by acting both on cells and bacteria.

Pediatrics, 2003 Jun, 111(6 Pt 1), e641 - 4
Response to immunization with measles, tetanus, and Haemophilus influenzae type b vaccines in children who have human immunodeficiency virus type 1 infection and are treated with highly active antiretroviral therapy; Melvin AJ et al.; OBJECTIVE: To assess the level of immunity to measles, tetanus, and Haemophilus influenzae type b (Hib) in previously immunized children who have human immunodeficiency virus (HIV) infection and were treated with highly active antiretroviral therapy (HAART) and to determine the response to reimmunization . METHODS: Retrospective review of clinical data from children who have HIV-1 infection and were treated with HAART . Children were included in the analysis when they had a history of immunizations before treatment with HAART; had specific immunoglobulin G levels to tetanus, measles, or Hib measured after starting HAART but before the receipt of additional immunizations; were reimmunized while on HAART; and had postimmunization immunoglobulin G levels available . RESULTS: Nineteen children (median age: 7 years; range: 3-14 years) who were treated with 3 to 5 drug HAART regimens for a median of 20 months (range: 8-37) met the criteria for at least 1 antigen and were included in this review . Fifteen (79%) of the 19 had plasma RNA levels <50 copies/mL . The median CD4% before HAART was 26% (range: 1-41) and at the time of immunization, 35% (range: 20-54) . Before reimmunization, 1 (5%) of 18 children had detectable antibody levels to measles, 6 (35%) of 17 had detectable antibody levels to tetanus, and 14 (78%) of 18 had detectable antibody levels to Hib . After immunization, 15 (83%) of 18, 10 (90%) of 11, and 3 (75%) of 4 seroconverted to measles, tetanus, and Hib, respectively . Antibody levels remained detectable after 1 year in the majority of children tested . CONCLUSIONS: Consideration should be given to readministering childhood immunizations to children who have HIV infection and are treated successfully with combination antiretroviral therapy.

EMBO J, 2003 Jun 2, 22(11), 2593 - 603
Crystal structure of tRNA(m1G37)methyltransferase: insights into tRNA recognition; Ahn HJ et al.; tRNA(m(1)G37)methyltransferase (TrmD) catalyzes the transfer of a methyl group from S-adenosyl-L- methionine (AdoMet) to G(37) within a subset of bacterial tRNA species, which have a G residue at the 36th position . The modified guanosine is adjacent to and 3' of the anticodon and is essential for the maintenance of the correct reading frame during translation . Here we report four crystal structures of TrmD from Haemophilus influenzae, as binary complexes with either AdoMet or S-adenosyl-L-homocysteine (AdoHcy), as a ternary complex with AdoHcy and phosphate, and as an apo form . This first structure of TrmD indicates that it functions as a dimer . It also suggests the binding mode of G(36)G(37) in the active site of TrmD and the catalytic mechanism . The N-terminal domain has a trefoil knot, in which AdoMet or AdoHcy is bound in a novel, bent conformation . The C-terminal domain shows structural similarity to trp repressor . We propose a plausible model for the TrmD(2)-tRNA(2) complex, which provides insights into recognition of the general tRNA structure by TrmD.

FEMS Immunol Med Microbiol, 2003 Jun 10, 37(1), 69 - 75
Infectious exacerbations of chronic obstructive pulmonary disease associated with respiratory viruses and non-typeable Haemophilus influenzae; Bandi V et al.; Infectious exacerbations of chronic obstructive pulmonary disease (COPD) have been reported to occur with both viral and bacterial pathogens . In this study, 35 exacerbations associated with the isolation of non-typeable Haemophilus influenzae from sputum were identified as part of a prospective longitudinal study . Samples from these patients were subjected to immunoassays to identify a new immune response to the homologous isolate of non-typeable H . influenzae to more accurately assess a bacterial etiology . These patients also were studied carefully for evidence of viral infection using viral culture, serology and polymerase chain reaction-based assays . Sixteen of 35 exacerbations (45.7%) were associated with evidence of acute viral infection and 11 of the 35 exacerbations (31.4%) were associated with the development of new serum IgG to homologous non-typeable H . influenzae . Overall, evidence of infection with a respiratory virus or non-typeable H . influenzae was seen in 24 of 35 exacerbations (68.6%) . No association between viral infection and immune response to non-typeable H . influenzae was observed, although a trend toward an immune response to non-typeable H . influenzae and absence of viral infection was seen . The results show that exacerbations in adults with COPD were associated with infection caused by virus alone, non-typeable H . influenzae alone, or virus and non-typeable H . influenzae simultaneously.

Arch Dis Child, 2003 Jun, 88(6), 536 - 9
Chloramphenicol or ceftriaxone, or both, as treatment for meningitis in developing countries?
Duke T, Michael A, Mokela D, Wal T, Reeder J.
AIMS: To determine in children with meningitis whether there is any difference in mortality and neurological sequelae using chloramphenicol as first line treatment, with a change to ceftriaxone if chloramphenicol resistance is shown in vitro, compared to using ceftriaxone as first line treatment, with a change to chloramphenicol if there is no evidence of in vitro resistance . METHODS: An observational study with a retrospective control group nested within a randomised trial of fluid management for bacterial meningitis where clinical care was standardised . Chloramphenicol is standard treatment for bacterial meningitis in Papua New Guinea . In the first 150 cases we used chloramphenicol and only changed treatment to ceftriaxone if chloramphenicol resistance for cerebrospinal fluid isolates was proved . After finding 20% of Haemophilus influenzae were resistant to chloramphenicol, and that most affected children had poor outcomes, we changed to an alternative strategy . In the next 196 cases first line treatment was ceftriaxone and treatment was changed to chloramphenicol if the isolated bacteria were found to be susceptible . RESULTS: When chloramphenicol was used as first line treatment for meningitis followed by ceftriaxone when in vitro resistance was shown, there was invariably a very poor outcome in chloramphenicol resistant disease (71% of children died or had severe neurological complications) . Using ceftriaxone as first line treatment was effective in reducing mortality and neurological sequelae from chloramphenicol resistant Haemophilus influenzae type (71% v 9%, relative risk 0.13; 95% CI 0.02 to 0.87; p = 0.013) . Changing to chloramphenicol if there was no evidence of in vitro resistance was less than half the cost of empirical use of ceftriaxone for a full course for all children with meningitis . CONCLUSIONS: Using a third generation cephalosporin as first line treatment is effective in dealing with the problem of poor outcomes from meningitis due to Haemophilus influenzae that is resistant to chloramphenicol, and a strategy of changing to chloramphenicol if in vitro susceptibility is shown will reduce the use of expensive third generation cephalosporins without comprising on clinical outcomes . This highlights the urgent need to reduce the costs of third generation cephalosporins, to improve bacteriological services in developing countries, and to introduce effective and affordable vaccines against H influenzae and Streptococcus pneumoniae.

N Y State Dent J, 2003 Mar, 69(3), 34 - 6
Haemophilus influenza cellulitis . A review and case report; Branca G et al.; Buccal cellulitis resulting from Haemophilus Influenzae type B (Hlb) is an uncommon yet potentially life-threatening illness that afflicts the facial soft tissues of the very young . Early recognition is essential for the effective treatment of this illness . A clinical case of Haemophilus Influenzae buccal cellulitis is presented, accompanied by a discussion of the presenting symptoms, diagnosis and treatment of this unusual childhood infection.

Infect Immun, 2003 Jun, 71(6), 3639 - 44
Complete sequence of the cap locus of Haemophilus influenzae serotype b and nonencapsulated b capsule-negative variants; Satola SW et al.; The complete capsule (cap) loci from three Haemophilus influenzae strains, one serotype b (Hib) and two nonencapsulated b capsule-negative variants, were sequenced . Two new open reading frames, hcsA and hcsB, were identified in region III and thought to be involved in postpolymerization modification of the capsule . The location of the cap locus in the Haemophilus influenzae chromosome was identified within section 97 of the Rd genome (chromosomal coordinates 1074542 to 1086327) and found to be the same for the Hib and two Hib(-) strains as well as some other encapsulated division I H . influenzae strains.

Infect Immun, 2003 Jun, 71(6), 3454 - 62
Nontypeable Haemophilus influenzae gene expression induced in vivo in a chinchilla model of otitis media; Mason KM et al.; The gram-negative bacterium nontypeable Haemophilus influenzae (NTHI) is the predominant pathogen in chronic otitis media with effusion and, with Streptococcus pneumoniae and Moraxella catarrhalis, is a causative agent of acute otitis media . To identify potential virulence determinants, bacterial gene expression was monitored by differential fluorescence induction during early disease progression in one specific anatomical niche of a chinchilla model of NTHI-induced otitis media . Genomic DNA fragments from NTHI strain 86-028NP were cloned upstream of the promoterless gfpmut3 gene . NTHI strain 86-028NP served as the host for the promoter trap library . Pools of 2,000 transformants were inoculated into the left and right middle ear cavities of chinchillas . Middle ear effusions were recovered by epitympanic tap at 24 and 48 h, and clones containing promoter elements that were induced in vivo and producing green fluorescent protein were isolated by two-color fluorescence-activated cell sorting . Insert DNA was sequenced and compared to the complete genome sequence of H . influenzae strain Rd . In a screen of 16,000 clones, we have isolated 44 clones that contain unique gene fragments encoding biosynthetic enzymes, metabolic and regulatory proteins, and hypothetical proteins of unknown function . An additional eight clones contain gene fragments unique to our NTHI isolate . Using quantitative reverse transcription-PCR, we have confirmed that 26 clones demonstrated increased gene expression in vivo relative to expression in vitro . These data provide insight into the response of NTHI bacteria as they sense and respond to the middle ear microenvironment during early events of otitis media.

Antimicrob Agents Chemother, 2003 Jun, 47(6), 1963 - 7
In vitro activities of telithromycin and 10 oral agents against aerobic and anaerobic pathogens isolated from antral puncture specimens from patients with sinusitis; Goldstein EJ et al.; A study of the comparative in vitro activity of telithromycin, a new ketolide, against 155 aerobic and 171 anaerobic antral sinus puncture isolates showed it to be active against a broad range of sinus pathogens . All pneumococci, including erythromycin-resistant strains, were susceptible to telithromycin at < or = 0.5 microg/ml; all Haemophilus influenzae and Eikenella corrodens strains were inhibited by < or = 4 microg of telithromycin/ml; all Moraxella spp . and beta-lactamase-producing Prevotella species strains were inhibited by < or = 0.25 and 0.5 microg of telithromycin/ml, respectively . Among all anaerobes tested, 94% (160 of 171 strains) were susceptible to < or = 4 microg of telithromycin/ml; however, 8 of 17 (47%) Fusobacterium strains, 2 Veillonella strains, and 1 Peptostreptococcus micros strain required > 4 microg of telithromycin/ml for inhibition . Telithromycin may offer a therapeutic alternative for sinus infections, including those due to erythromycin-resistant pneumococci.

Antimicrob Agents Chemother, 2003 Jun, 47(6), 1875 - 81
Antimicrobial resistance in Haemophilus influenzae and Moraxella catarrhalis respiratory tract isolates: results of the Canadian Respiratory Organism Susceptibility Study, 1997 to 2002; Zhanel GG et al.; A total of 7,566 unique patient isolates of Haemophilus influenzae and 2,314 unique patient isolates of Moraxella catarrhalis were collected between October 1997 and June 2002 from 25 medical centers in 9 of the 10 Canadian provinces . Among the 7,566 H . influenzae isolates, 22.5% produced beta-lactamase, while 92.4% of the 2,314 M . catarrhalis isolates produced beta-lactamase . The incidence of beta-lactamase-producing H . influenzae isolates decreased significantly over the 5-year study period, from 24.2% in 1997-1998 to 18.6% in 2001-2002 (P < 0.01) . The incidence of beta-lactamase-producing M . catarrhalis isolates did not change over the study period . The overall rates of resistance to amoxicillin and amoxicillin-clavulanate for H . influenzae were 19.3 and 0.1%, respectively . The rank order of cephalosporin activity based on the MICs at which 90% of isolates were inhibited (MIC(90)s) was cefotaxime > cefixime > cefuroxime > cefprozil > cefaclor . On the basis of the MICs, azithromycin was more active than clarithromycin (14-OH clarithromycin was not tested); however, on the basis of the NCCLS breakpoints, resistance rates were 2.1 and 1.6%, respectively . Rates of resistance to other agents were as follows: doxycycline, 1.5%; trimethoprim-sulfamethoxazole, 14.2%; and chloramphenicol, 0.2% . All fluoroquinolones tested, including the investigational fluoroquinolones BMS284756 (garenoxacin) and ABT-492, displayed potent activities against H . influenzae, with MIC(90)s of < or = 0.03 microg/ml . The MIC(90)s of the investigational ketolides telithromycin and ABT-773 were 2 and 4 microg/ml, respectively, and the MIC(90) of the investigational glycylcycline GAR-936 (tigecycline) was 4 microg/ml . Among the M . catarrhalis isolates tested, the resistance rates derived by using the NCCLS breakpoint criteria for H . influenzae were <1% for all antibiotics tested except trimethoprim-sulfamethoxazole (1.5%) . In summary, the incidence of beta-lactamase-positive H . influenzae strains in Canada is decreasing (18.6% in 2001-2002), while the incidence of beta-lactamase-positive M . catarrhalis strains has remained constant (90.0% in 2001-2002).

Antimicrob Agents Chemother, 2003 Jun, 47(6), 1790 - 7
Susceptibilities to levofloxacin in Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis clinical isolates from children: results from 2000-2001 and 2001-2002 TRUST studies in the United States; Karlowsky JA et al.; Among respiratory tract isolates of Streptococcus pneumoniae from children, resistance to penicillins, cephalosporins, macrolides, and trimethoprim-sulfamethoxazole (SXT) increases on an annual basis . Pediatric patients who do not respond to conventional therapy for respiratory tract infections someday may be treated with fluoroquinolones . In this study, MICs of beta-lactams, azithromycin, SXT, and levofloxacin were determined and interpreted by using NCCLS guidelines for isolates of S . pneumoniae (2,834 from children and 10,966 from adults), Haemophilus influenzae (629 from children and 2,281 from adults), and Moraxella catarrhalis (389 from children and 1,357 from adults) collected during the 2000-2001 and 2001-2002 respiratory illness seasons in the United States as part of the ongoing TRUST surveillance studies . Rates of resistance to penicillin, azithromycin, and SXT were > or = 7.5% higher among patients < or = 4 years old than among patients 5 to 10, 11 to 17, and > or = 18 years old in both the 2000-2001 and the 2001-2002 respiratory illness seasons . Levofloxacin resistance was detected in 2 of 2,834 isolates (0.07%) from patients <18 years old . Levofloxacin MICs of 0.25 to 1 micro g/ml accounted for 99.6, 99.5, 99.3, 99.7, 98.4, and 98.0% of isolates from patients < 2, 2 to 4, 5 to 10, 11 to 17, 18 to 64, and > 64 years old . Multidrug resistance was twice as common among patients < or = 4 years old (25.3%) as among patients 5 to 10 years old (13.7%), 11 to 17 years old (11.9%), 18 to 64 years old (12.1%), and > 64 years old (12.4%) . The most common multidrug resistance phenotype in S . pneumoniae isolates for all age groups was resistance to penicillin, azithromycin, and SXT (70.3 to 76.6%) . For H . influenzae and M . catarrhalis isolates from patients < 2, 2 to 4, 5 to 10, 11 to 17, 18 to 64, and > 64 years old, levofloxacin MICs at which 90% of the isolates were inhibited were 0.015 and 0.03 to 0.06 microg/ml, respectively, in the 2000-2001 and 2001-2002 respiratory illness seasons . In the 2000-2001 and 2001-2002 respiratory illness season surveillance studies in the United States, 99.9% of pediatric isolates of S . pneumoniae were susceptible to levofloxacin . If fluoroquinolones become a treatment option for pediatric patients, careful monitoring of fluoroquinolone susceptibilities will be increasingly important in future surveillance studies.

Can J Vet Res, 2003 May, 67(2), 146 - 50
Naturally-farrowed, artificially-reared pigs as an alternative model for experimental infection by Haemophilus parasuis; Oliveira S et al.; The use of naturally-farrowed, artificially-reared piglets as an alternative model to study Haemophilus parasuis infections was evaluated . Two trials were performed in order to evaluate the proposed model . In trial 1, animals were vaccinated and challenged with H . parasuis . Results showed that the proposed model was effectively used to evaluate protective immunity against this organism . In trial 2, animals were challenged with different doses of H . parasuis . Results showed that the severity of clinical signs and lesions tended to increase with higher doses . The reproduction of clinical signs and lesions characteristic of H . parasuis systemic infection was successful in both trials, proving that this model is a viable alternative to specific-pathogen free and cesarean-derived, colostrum-deprived pigs.

Biochim Biophys Acta, 2003 May 30, 1648(1-2), 203 - 9
The molecular class C acid phosphatase of Chryseobacterium meningosepticum (OlpA) is a broad-spectrum nucleotidase with preferential activity on 5'-nucleotides; Passariello C et al.; The olpA gene of Chryseobacterium meningosepticum, encoding a molecular class C phosphatase, was cloned and expressed in Escherichia coli . The gene encodes a 29-kDa polypeptide containing an amino-terminal signal peptide typical of bacterial membrane lipoproteins . Expression in E . coli results in a functional product that mostly partitions in the outer membrane . A secreted soluble OlpA derivative (sOlpA) lacking the N-terminal cysteine residue for lipid anchoring was produced in E . coli and purified by means of two steps of ion exchange chromatography . Analysis of the kinetic parameters of sOlpA with several organic phosphoesters revealed that the enzyme was able to efficiently hydrolyze nucleotide monophosphates, with a strong preference for 5'-nucleotides and for 3'-AMP . The enzyme was also able to hydrolyze sugar phosphates and beta-glycerol phosphate, although with a lower efficiency, whereas it was apparently inactive against nucleotide di- and triphosphates, diesters, and phytate . OlpA, therefore, can be considered a broad-spectrum nucleotidase with preference for 5'-nucleotides . Its functional behaviour exhibits differences from that of the Haemophilus influenzae OMP P4 lipoprotein, revealing functional heterogeneity among phosphatases of molecular class C.

Carbohydr Res, 2003 May 23, 338(11), 1223 - 8
Structural analysis of the lipooligosaccharide from the commensal Haemophilus somnus strain 1P; Cox AD et al.; The structure of the lipooligosaccharide (LOS) from the commensal Haemophilus somnus strain 1P was elucidated . The structure of the O-deacylated LOS was established by monosaccharide analysis, NMR spectroscopy and mass spectrometry . The following structure for the O-deacylated LOS was determined on the basis of the combined data from these experiments . {chemical structure: see text} In the structure Kdo is 3-deoxy-D-manno-octulosonic acid, Hep is L-glycero-D-manno-heptose and lipid A-OH refers to O-deacylated Lipid A . The elucidation of this structure has increased our understanding of the relationship between the variability in LOS structure and the pathogenic potential of this organism . Specifically, the inability of this commensal strain to sialylate its LOS suggests that LOS sialylation could be a crucial virulence factor for H . somnus.

Vaccine, 2003 Jun 2, 21(19-20), 2273 - 87
Development and clinical testing of multivalent vaccines based on a diphtheria-tetanus-acellular pertussis vaccine: difficulties encountered and lessons learned; Capiau C et al.; The widespread use of whole-cell pertussis vaccines in the second half of the 20th century have reduced the incidence of the disease significantly . However, in some countries, concerns about the reactogenicity and potential neurological damage associated with whole-cell vaccines led to a decrease in vaccine acceptance and an increase in morbidity and mortality of pertussis in several countries . This prompted the development of less reactogenic acellular pertussis vaccines combined with diphtheria and tetanus toxoids, initially in Japan and later in other countries . In Europe, the improved diphtheria, tetanus and acellular pertussis (DTPa) vaccine was first introduced in March 1994.The pertussis component of this DTPa vaccine, developed by Glaxo SmithKline, consists of pertussis toxoid, filamentous haemagglutinin and pertactin . The vaccine is well tolerated, with a lower incidence of adverse reactions than after administration of whole-cell vaccines.The long-lasting efficacy and effectiveness of DTPa vaccines have been extensively documented and these are now the cornerstone of a large range of combined vaccines including DTPa-hepatitis B (HBV), DTPa-inactivated polio (IPV) and DTPa-HBV-IPV . A lyophilised Haemophilus influenzae type b (Hib) vaccine can be reconstituted with all of these liquid combinations . The introduction of well-tolerated and efficacious DTPa vaccines and their more polyvalent combinations has improved the acceptance and simplified the implementation of childhood immunisation . This paper is a review of the technical and scientific difficulties encountered and the lessons learned over the 10-year period that it took to develop and introduce six multivalent vaccines using the Glaxo SmithKline DTPa as a building block.

Diagn Microbiol Infect Dis, 2003 May, 46(1), 55 - 61
Five-year analysis of Haemophilus influenzae isolates with reduced susceptibility to fluoroquinolones: prevalence results from the SENTRY antimicrobial surveillance program; Biedenbach DJ et al.; The appearance of resistance or reduced susceptibility to fluoroquinolones among Hemophilus influenzae has been documented for nearly a decade . Over this time, the use of fluoroquinolones for the treatment of respiratory infections including commonly isolated bacterial causes of community-acquired infections has markedly increased . The documentation of resistance to fluoroquinolones among Streptococcus pneumoniae and H . influenzae has also become more prevalent as measured by peer-reviewed publications . During 1997-2001, a total of 11,355 H . influenzae isolates were tested by reference broth microdilution methods from strains collected by the SENTRY Antimicrobial Surveillance Program (American and European medical centers) . Strains with reduced susceptibility to fluoroquinolones (RSF) were detected during all five study years at an overall rate of 0.15% . Among the tested compounds, sitafloxacin (MIC(50,) 0.03 microg/ml) was the most potent agent against the RSF strains, followed by gemifloxacin (0.12 microg/ml) > garenoxacin = grepafloxacin = levofloxacin = moxifloxacin = trovafloxacin (0.5 microg/ml) > ciprofloxacin = sparfloxacin (1 microg/ml) . Gene sequencing of the quinolone resistance determining region and epidemiologic typing of 30 RSF isolates showed diverse mutational events in gyr A and par C and multiple pulsed-field gel electrophoresis (PFGE) patterns among strains that was not consistent with clonal dissemination . Continued surveillance by global or national networks should continue to monitor for H . influenzae isolates that are refractory to fluoroquinolone therapy.

J Microbiol Immunol Infect, 2003 Mar, 36(1), 69 - 71
Acute epiglottitis caused by Haemophilus influenzae type b: a case report; Jiang JH et al.; Acute epiglottitis is an inflammatory, edematous disease of the epiglottis and adjacent structures, usually caused by Haemophilus influenzae type b . It is a life-threatening condition, occurring mainly in childhood . There have never been any reports of this condition in Taiwan . We report a case of 4-year-old boy who presented with characteristics of systemic illness combined with respiratory distress on arrival at the emergency room . His mouth was open and his neck was hyperextended . The diagnosis of epiglottitis was established on the basis of physical examination, lateral neck x-ray, and the finding of an enlarged, swollen, erythematous epiglottis on flexible fiberoptic laryngoscopy . Urine latex agglutination test for H . influenzae type b was positive and a blood culture grew H . influenzae type b . He was treated with cefotaxime and did not require intubation.

J Microbiol Immunol Infect, 2003 Mar, 36(1), 41 - 6
Septic arthritis in children: relationship of causative pathogens, complications, and outcome; Wang CL et al.; This retrospective study investigated the causative pathogens, complications, and outcome of 58 children who were hospitalized for septic arthritis at a tertiary care hospital in southern Taiwan from July 1988 to December 2000 . The mean age was 3 years (range, 12 days-16 years) . The males/females ratio was 1.2:1 . Ninety percent of the cases involved lower extremities (knee, hip, and ankle) with the hip being the most common site of infection (54%) . Joint pain (81%) was the most common clinical presentation, followed by fever (74%), local warmness and swelling (72%), and limitation of motion (64%) . Erythrocyte sedimentation rate was elevated (> or = 20 mm/h) initially in 89% of the cases . The predominant causative organism was Staphylococcus aureus (43%, 25/58), 6 isolates of which were methicillin-resistant, followed by coagulase-negative Streptococcus (6), Streptococcus pneumoniae (3), Salmonella spp . (3), Haemophilus influenzae type b (2), and group B Streptococcus (2) . The concomitant complications of septic arthritis were sepsis (9%, 5/58) and meningitis (2%, 1/58) . Ten patients had sequelae, including limitation of motion (6), limping gait (2), limb-length discrepancy (1), and abnormalities of bone growth (1) . This study found that S . aureus was the most common infecting microorganism in septic arthritis in children . Septic arthritis with concomitant osteomyelitis and infection due to methicillin-resistant S . aureus was associated with a significantly increased risk of sequelae (relative risk, 46.4, 95% CI, 2.9-748.8; relative risk, 16 . 2, 95% CI, 1.3-204.9, respectively).

Lancet, 2003 May 3, 361(9368), 1523 - 4
Antibody to Haemophilus influenzae type b after routine and catch-up vaccination; Trotter CL et al.; Since 1999, the number of cases of Haemophilus influenzae type b (Hib) disease in the UK has risen . We investigated the role of population immunity in this change by testing more than 2600 serum samples from children aged 1-15 years . After the introduction of the routine Hib conjugate vaccination programme for infants, median antibody titres rose significantly in 1-year-olds . Individuals who received their first dose of vaccine at age 1-4 years in the original catch-up campaign initially had much higher concentrations of antibody than those who had been immunised in infancy . A second catch-up campaign in children aged 6 months to 4 years should be highly effective in boosting immunity and reducing disease in the short term.

Lancet, 2003 May 3, 361(9368), 1521 - 3
Risk of vaccine failure after Haemophilus influenzae type b (Hib) combination vaccines with acellular pertussis; McVernon J et al.; An increase in invasive Hib disease incidence in the UK has coincided with the distribution of combination vaccines that contain acellular pertussis (DTaP-Hib) . These vaccines have been associated with reduced immunogenicity of the Hib component, although there is little agreement on the clinical relevance of this finding . We retrospectively compared vaccine formulations given to fully vaccinated Hib cases with those administered to fully immunised age-matched controls using conditional logistic regression . More cases than controls received all three doses of their infant primary course as DTaP-Hib, compared with two or three doses of another Hib vaccine (conditional odds ratio 6.77 {95% CI 3.26-14.07}).

Commun Dis Public Health, 2003 Apr, 6(1), 55 - 8
Rising incidence of Haemophilus influenzae type b disease in England and Wales indicates a need for a second catch-up vaccination campaign; Trotter CL et al.; The incidence of invasive Haemophilus influenzae type b (Hib) disease in the UK fell rapidly following the introduction of routine vaccination in 1992 and the implementation of a catch-up campaign in children under 4 years old in 1992-93 . However, since 1999 the number of cases of Hib has been increasing, and in 2002 there were 134 cases in 0-4 year olds (266 in all ages) . While still much less than the prevaccine burden of disease (over 800 cases a year in 0-4 year olds), this increase in incidence is worrying and has sparked a range of detailed investigations . In February 2003, the Department of Health announced a second catch-up campaign offering all children between 6 months and 4 years a further dose of Hib vaccine . The epidemiology of Hib disease in England and Wales between 1990 and 2002 is reviewed here to provide a context for this public health response.

J Clin Microbiol, 2003 May, 41(5), 2080 - 3
Pgi genotyping is a surrogate for serotyping of encapsulated Haemophilus influenzae; Anyanwu JN et al.; Study of the epidemiology of invasive infections caused by encapsulated Haemophilus influenzae has been complicated by the poor sensitivity and specificity of the serologic assays used to identify specific capsular polysaccharides . The population structure of these bacteria is highly clonal, however, and serotype is highly correlated with other genetic characteristics . We sought to determine if alleles of the highly conserved phosphoglucose isomerase (pgi) gene correspond to the serotypes of encapsulated H . influenzae strains . pgi alleles of 52 well-characterized encapsulated H . influenzae isolates were amplified by PCR, sequenced, and compared to one another and to additional previously reported H . influenzae pgi alleles . Overall, 83% of the strains possessed pgi alleles associated with the major serotype a, b, e, and f clonotypes that cause the most invasive disease in the United States . Six strains (four type a and two type f) had unusual pgi alleles, which suggested that these strains belonged to less common clonotypes of encapsulated bacteria or were actually nontypeable strains . pgi genotyping may provide a simple and stable surrogate for capsular serotyping . Further studies correlating pgi typing with the expression of capsule are likely to increase our understanding of the epidemiology and pathogenesis of these infections.

J Biol Chem, 2003 Jul 25, 278(30), 27811 - 9 Epub 2003 May 06.
Transforming growth factor-beta-Smad signaling pathway negatively regulates nontypeable Haemophilus influenzae-induced MUC5AC mucin transcription via mitogen-activated protein kinase (MAPK) phosphatase-1-dependent inhibition of p38 MAPK; Jono H et al.; In contrast to the extensive studies on the role of transforming growth factor-beta (TGF-beta) in regulating cell proliferation, differentiation, and apoptosis over the past decade, relatively little is known about the exact role of TGF-beta signaling in regulating host response in infectious diseases . Most of the recent studies have suggested that TGF-beta inhibits macrophage activation during infections with pathogens such as Trypanosoma cruzi and Leishmania, thereby favoring virulence . In certain situations, however, there is also evidence that TGF-beta has been correlated with enhanced resistance to microbes such as Candida albicans, thus benefiting the host . Despite these distinct observations that mainly focused on macrophages, little is known about how TGF-beta regulates host primary innate defensive responses, such as up-regulation of mucin, in the airway epithelial cells . Moreover, how the TGF-beta-Smad signaling pathway negatively regulates p38 mitogen-activated protein kinase (MAPK), a key pathway mediating host response to bacteria, still remains largely unknown . Here we show that nontypeable Haemophilus influenzae, a major human bacterial pathogen of otitis media and chronic obstructive pulmonary diseases, strongly induces up-regulation of MUC5AC mucin via activation of the Toll-like receptor 2-MyD88-dependent p38 path-way . Activation of TGF-beta-Smad signaling, however, leads to down-regulation of p38 by inducing MAPK phophatase-1, thereby acting as a negative regulator for MUC5AC induction . These studies may bring new insights into the novel role of TGF-beta signaling in attenuating host primary innate defensive responses and enhance our understanding of the signaling mechanism underlying the cross-talk between TGF-beta-Smad signaling pathway and the p38 MAPK pathway.

Diagn Microbiol Infect Dis, 2003 Apr, 45(4), 279 - 85
Pathogen of occurrence and susceptibility patterns associated with pneumonia in hospitalized patients in North America: results of the SENTRY Antimicrobial Surveillance Study (2000); Hoban DJ et al.; Antimicrobial selection for patients diagnosed with pneumonia is a major therapeutic challenge and dilemma to the clinical practitioner . In the community setting, patients usually receive empiric oral therapy based upon multiple patient risk factors and locally prevalent pathogen susceptibilities . For patients admitted to the hospital with pneumonia, or who acquire pneumonia while in the hospital, therapy can be initially empiric and then become directed once culture and susceptibility results are known . The SENTRY Antimicrobial Surveillance Program since 1997, has monitored pathogen frequency and antimicrobial susceptibilities in hospitalized patients with pneumonia in North America . In this Study 2,712 pathogens were studied from 30 medical centers (25 in the United States and 5 in Canada) . Over 30 species of organisms were recovered with Staphylococcus aureus comprising 28.0% of all isolates and with four other species (Pseudomonas aeruginosa 20.0%, Streptococcus pneumoniae 9.1%, Klebsiella spp . 7.5% and Haemophilus influenzae 7.3%) constituted 71.9% of isolates submitted . Methicillin (oxacillin)-resistant S . aureus accounted for 43.7% of all S . aureus isolates . Antimicrobials demonstrating significant (>80%) activity against S . aureus were: chloramphenicol (81.6%), tetracycline (91.4%), rifampin (96.4%) and quinupristin/dalfopristin (99.7%); and no isolate was resistant to glycopeptides or linezolid . North American isolates of P . aeruginosa were most susceptible to amikacin (93.7%) > tobramycin (90.2%) > meropenem (89.1%) > imipenem = piperacillin/tazobactam (85.6%) > piperacillin (82.1%) > cefepime (80.5%) . Overall, 32.1% of S . pneumoniae were penicillin non-susceptible while erythromycin susceptibility was only 74.8% . Fluoroquinolones and recent generation cephalosporins retained excellent activity (gatifloxacin {99.2%} > levofloxacin = cefepime {98.8%} > ceftriaxone {97.2%}) against S . pneumoniae . Klebsiella spp . were 100.0% susceptible to the carbapenems (meropenem and imipenem) but extended spectrum beta-lactamases were detected at a rate of 5.4% . The beta-lactamase-positive rate in H . influenzae was 28.6% in North America (71.4% ampicillin-susceptible) . The SENTRY Antimicrobial Surveillance Program continues to identify important North American patterns of pathogen frequency and resistance . Additionally, the provision of multi-year longitudinal data and associated reports allow for comparisons, which function as critical tools for effective patient management and antimicrobial interventions.

Pediatrics, 2003 May, 111(5 Pt 1), 925 - 32
Physician knowledge of catch-up regimens and contraindications for childhood immunizations; Cohen NJ et al.; OBJECTIVES: To determine physician success at designing catch-up regimens for children delayed in immunizations and physician knowledge regarding contraindications to immunization . METHODS: A self-administered survey was completed by pediatricians, general practitioners, and family practitioners in Cook County, Illinois . Surveys included 6 open-ended vignettes describing hypothetical children delayed in immunization for whom participants were asked to design catch-up regimens . Bivariate and multivariate logistic regression were used to determine predictors of correct response . The surveys also inquired about management of scenarios that might be perceived as contraindications to immunize with the Haemophilus influenzae type b or measles-mumps-rubella vaccines . RESULTS: The mean score of correct responses was 1.83 of a possible 6.0 . Almost one third of respondents answered all 6 vignettes incorrectly . The proportion of incorrect responses was high for all 6 vignettes (39%-86%), but higher for questions that addressed the immunization of children older than 12 months . Errors in vaccine administration were most commonly attributed to omitted vaccines, with varicella-zoster vaccine and pneumococcal conjugate vaccine omitted most frequently . Pediatricians were >4 times more likely to answer correctly than were family practitioners . Participants in the Vaccines for Children (VFC) program were more than twice as likely to answer correctly than were non-VFC providers . Knowledge of contraindications was inconsistent, particularly for measles-mumps-rubella vaccine . CONCLUSIONS: Childhood vaccine providers have substantial knowledge deficits of recommended immunization schedules and vaccine contraindications that may contribute to missed opportunities to immunize . Pediatricians and participants in the VFC program were more successful at designing catch-up regimens for children with immunization delay.

Commun Dis Intell, 2003, 27(1), 1 - 78
Australia's notifiable diseases status, 2001: annual report of the National Notifiable Diseases Surveillance System; Blumer C et al.; In 2001 there were 104,187 notifications of communicable diseases in Australia reported to the National Notifiable Diseases Surveillance System (NNDSS) . The number of notifications in 2001 was an increase of 16 per cent of those reported in 2000 (89,740) and the largest annual total since the NNDSS commenced in 1991 . In 2001, nine new diseases were added to the list of diseases reported to NNDSS and four diseases were removed . The new diseases were cryptosporidiosis, laboratory-confirmed influenza, invasive pneumococcal disease, Japanese encephalitis, Kunjin virus infection, Murray Valley encephalitis virus infection, anthrax, Australian bat lyssavirus, and other lyssaviruses (not elsewhere classified) . Bloodborne virus infections remained the most frequently notified disease (29,057 reports, 27.9% of total), followed by sexually transmitted infections (27,647, 26.5%), gastrointestinal diseases (26,086, 25%), vaccine preventable diseases (13,030 (12.5%), vectorborne diseases (5,294, 5.1%), other bacterial infections (1,978, 1.9%), zoonotic infections (1,091, 1%) and four cases of quarantinable diseases . In 2001 there were increases in the number of notifications of incident hepatitis C, chlamydial infections, pertussis, Barmah Forest virus infection and ornithosis . There were decreases in the number of notifications of hepatitis A, Haemophilus influenzae type b infections, measles, rubella, Ross River virus infections and brucellosis . This report also summarises data on communicable diseases from other surveillance systems including the Laboratory Virology and Serology Reporting Scheme and sentinel general practitioner schemes . In addition, this report comments on other important developments in communicable disease control in Australia in 2001.

Nihon Kokyuki Gakkai Zasshi, 2003 Feb, 41(2), 89 - 94
{Clinical properties of community-acquired pneumonia in patients with asthma}; To M et al.; PURPOSE: Many reports were found on the clinical properties of community-acquired pneumonia . The clinical properties of community-acquired pneumonia in patients with asthma have not been elucidated, and we therefore investigated such properties . MATERIALS AND METHODS: Asthmatic patients who required hospitalization for community-acquired pneumonia from the beginning of 1989 through the end of 2001 were enrolled in this study . We performed the study in a retrospective manner . Patients were divided into two groups based on severity of their asthma (mild to moderate asthma vs severe asthma), and we studied the clinical properties of the pneumonia . RESULT: No significant difference was seen in body temperature, white blood cell counts, or CRP value on admission between the two groups . No significant difference was seen in the resolving period of the pneumonia . The frequency of common pathogens (Streptococcus pneumoniae + Haemophilus influenzae) was lower in patients with severe asthma . Asthmatic patients not taking daily oral corticosteroids were divided into two groups based on whether or not they were using a inhaled corticosteroid, and we examined the frequency of pathogendetection . The percentage of common pathogens was almost the same in the two groups . CONCLUSION: The frequency of common pathogens was lower in patients with severe asthma than in those with mild to moderate asthma . This fact is worth considering when empiric therapy for pneumonia is performed in patients with asthma . Inhaled corticosteroid therapy seems to have no influence on the pathogens of pneumonia in patients with asthma.

J Clin Pathol, 2003 May, 56(5), 344 - 7
National surveillance programme on susceptibility patterns of respiratory pathogens in South Africa: moxifloxacin compared with eight other antimicrobial agents; Liebowitz LD et al.; AIMS: The susceptibility patterns of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Klebsiella pneumoniae, and Streptococcus pyogenes isolated from specimens submitted to 12 private laboratories in South Africa were determined . METHODS: Minimum inhibitory concentration (MIC) determinations were performed on the isolates in the microbiology laboratory at Tygerberg Hospital according to the recommendations of the National Committee for Clinical Laboratory Standards (NCCLS) . RESULTS: According to the NCCLS breakpoints, 24% of 729 S pneumoniae isolates were sensitive, 30% intermediate, and 46% resistant to penicillin . Rates of macrolide resistance were high, with 61% of the pneumococci being resistant to clarithromycin and azithromycin . Co-trimoxazole resistance was also high, with 28% of pneumococcal strains being sensitive, 21% intermediate, and 51% resistant . beta Lactamase was produced by 7% of 736 H influenzae isolates and 91% of 256 M catarrhalis isolates . The quinolones, moxifloxacin and levofloxacin, were universally active against all isolates tested, which included S pneumoniae, H influenzae, M catarrhalis, K pneumoniae, and S pyogenes . CONCLUSIONS: Haemophilus influenzae and S pneumoniae were the most commonly isolated organisms . Resistance to penicillin was one of the highest reported in the world (76%) in S pneumoniae, as was macrolide resistance in pneumonocci, although surprisingly, only 14% of S pyogenes were resistant . The quinolones, moxifloxacin and levofloxacin, were active against all organisms tested, including the penicillin and macrolide resistant strains and moxifloxacin was more active than levofloxacin against pneumococci.

Int J Infect Dis, 2002 Dec, 6(4), 288 - 93
Prospective epidemiologic survey of patients with community-acquired pneumonia requiring hospitalization in Switzerland; Garbino J et al.; BACKGROUND: Community-acquired pneumonia (CAP) is a common problem and the principal infection requiring hospitalization, but its treatment is complicated by the difficulty in microbiological diagnosis and the increasing incidence of antibiotic resistance among respiratory pathogens . The purpose of this paper is to present the main epidemiologic features of patients with CAP requiring hospitalization in our country . METHODS: We enrolled three hundred and eighteen adult patients with CAP requiring hospitalization in seven large medical centers in Switzerland during two winter periods . The patients' mean age was 70.4 years . This study describes the epidemiology of these patients . Clinical, radiologic and microbiological evaluations were performed at study entry during treatment, and at 4 weeks post-therapy . For microbiological diagnostic purposes, sputum culture, throat swab culture, PCR, blood cultures, Legionella urinary antigen and serologic evaluations were also performed . RESULTS: Despite the higher mean age, the overall mortality rate was 8%, lower than in other comparable studies . The most common underlying diseases present at study entry were cardiac failure (23%), chronic obstructive pulmonary disease (20%), renal failure (15%), and diabetes (12%); 40% of the patients were smokers . Although dyspnea, cough and positive pulmonary auscultation findings were present in about 90% of patients, fever >38 degrees C was present in only 64% . The most frequently isolated respiratory pathogens were Streptococcus pneumoniae (12.6%), Haemophilus influenzae (6%), Staphylococcus aureus (1.6%), and Moraxella catarrhalis (1.6%) . Atypical pathogens were frequently found, with the following distribution: Mycoplasma pneumoniae, 7.5%; Chlamydia pneumoniae, 5.3%; and Legionella pneumophila, 4.4% . The mean duration between onset of symptoms and hospital admission was 4.8 days, and the mean treatment duration was 12.1 days . Two weeks after the start of therapy, although clinical symptoms were absent, radiologic infiltrates were still present in 24% of patients . CONCLUSIONS: The microbiological diagnosis in CAP can be established in only about 50% of cases with the combination of several diagnostic tools . Epidemiologic surveys of CAP should be performed on a regular basis, regionally, as a way to improve the management of these infections.

J Coll Physicians Surg Pak, 2003 Apr, 13(4), 207 - 9
Seasonal paradox in acute meningitis at Nawabshah; Shaikh S et al.; OBJECTIVE: To assess the reasons for paradoxical increase in cases of meningitis in the summer of the year 2001 . DESIGN: An observational study . PLACE AND DURATION OF STUDY: Medical Unit III, Peoples Medical College Hospital, Nawabshah, Pakistan, from 1st January, 2001 to 31st December, 2001 . SUBJECTS AND METHODS: A total number of 229 cases of meningitis were admitted throughout the year with typical clinical features or CSF findings . Maximum number (179) was noted from April to July, which was alarming . The reason for this change was assessed in these cases . RESULTS: One hundred and seventy-nine cases of meningitis presented from 1st April to 31st July 2001 . They were predominantly males n= 107 (59.7%) with age range of 15 to 64 years . Presenting symptoms were fever and headache in all and altered sensorium in 160 (89%) cases with positive signs of meningeal irritation in 175 (97.8%) . Cerebrospinal fluid examination revealed altered biochemistry and neutrophil leucocytosis in all cases . Organism was identified in 139 (77.65%) cases on gram staining . Culture was done only in 61 (34.07%) . Predisposing factors were smoking in 93 (52%) and diabetes in 17 (9.5%) cases . Five female patients were pregnant . CONCLUSION: Common organisms detected were gram positive diplococci (Streptococcus pneumoniae) in 83 (59.7%), gram negative diplococci (Nisseria meningitides) in 51 (36.7%) and Haemophilus influenzae in 5 (3.6%) . As the frequency of isolates were approximately same as in other seasons, the increased in meningitis cases during summer was probably due to hot and dry weather causing low absolute humidity.

Arch Dis Child, 2003 May, 88(5), 379 - 83
Immunologic memory in Haemophilus influenzae type b conjugate vaccine failure; McVernon J et al.; AIMS: To compare the convalescent antibody response to invasive Haemophilus influenzae type b (Hib) disease between conjugate vaccine immunised and unimmunised children, to look for evidence of priming for immunologic memory . METHODS: Unmatched case-control study in the UK and Eire 1992-2001 and Victoria, Australia 1988-1990 . A total of 93 children were identified as having invasive Hib disease following three doses of conjugate vaccine in infancy through post licensure surveillance throughout the UK and Eire; 92 unvaccinated children admitted to an Australian paediatric hospital with invasive Hib disease were used as historical controls . Convalescent serum was taken for measurement of Hib antibody concentration, and clinical information relating to potential disease risk factors was collected . The geometric mean concentrations of convalescent Hib antibodies were compared between immunised and unimmunised children, using raw and adjusted data . RESULTS: Hib conjugate vaccine immunised children had higher serum Hib antibody responses to disease (geometric mean concentration (GMC) 10.81 microg/ml (95% CI 6.62 to 17.66) than unimmunised children (1.06 microg/ml (0.61 to 1.84)) (p < 0.0001) . However, following adjustment for the significant confounding influences of age at presentation and timing of serum collection, a difference persisted only in children presenting with meningitis (vaccinated GMC 3.78 microg/ml (2.78 to 5.15); unvaccinated GMC 1.48 microg/ml (0.90 to 2.21); p = 0.003) . CONCLUSIONS: Higher antibody responses to invasive Hib disease in vaccinated children with meningitis reflect priming for immunologic memory by the vaccine . Although a majority of children in the UK are protected from Hib disease by immunisation, the relative roles of immunologic memory and other immune mechanisms in conferring protection remain unclear.

Arch Dis Child, 2003 May, 88(5), 375 - 8
Group B streptococcal conjugate vaccines; Baker CJ et al.; Linkage of bacterial capsular polysaccharides to proteins to create conjugate vaccines has had a dramatic impact on the health of children . Although unconjugated polysaccharides are poorly immunogenic in infants and some older children and adults, their covalent coupling with proteins stimulates T cell dependent antigenic recognition that profoundly enhances immunogenicity . In the decade since the introduction and widespread use of Haemophilus influenzae type b polysaccharide conjugate vaccines in the United States, invasive H influenzae infections have become a rarity in childhood . Similarly, the conjugation of polysaccharides of Streptococcus pneumoniae to a derivative of diphtheria toxoid and the addition of pneumococcal conjugate vaccine to infant immunisation schedules carries with it promise for a similar decline in the incidence of invasive pneumococcal disease in paediatric patients.

Mymensingh Med J, 2003 Jan, 12(1), 36 - 40
Resistance pattern of bacteria isolated from acute respiratory tract infection (ARI) cases; Mahmud MC et al.; Out of 350 acute respiratory tract infection (ARI) patients 110 (31.4%) were positive on culture . Among then (35.5%) yielded from upper respiratory tract infection (URI) and 64.5% yielded from lower respiratory tract infection (LRI) . Predominant bacterial isolates from URI were Staph aureus (12.4%) and Strepto . pyogen (9.8%) whereas predominant agent from LRI were Strepto . pneumoniae (14.7%) and Haemophilus influenzae (8.6%) . Capsular typing by polymerase chain reaction (PCR) and type specific antisera revealed that 64.7% of the isolates were type-b and rest were non-b . The most frequent resistance pattern of H . influenzae was found resistant to penicillin-ampicillin (64.7%) followed by SMX-TMP (14.7%) and tetracycline (5.9%) . Among penicillin-ampicillin resistant strains, 81.8% were beta lectamase positive and 18.2% were beta lectamase negative . Among beta lectamase positive strains 66.7% were capsular type-b and 33.3% were non-b, had MICS < or = 8 microgram/ml . and < or = 4 microgram/ml . respectively . 5.9% strains of Strepto . pneumoniae was found resistant to SMX-TMP, 5.9% to penicillin-ampicillin and 2.9% to cephalexin . Common resistance pattern of Staph aureus was penicillin-ampicillin (60%), SMX-TMP (37.2%) and tetracycline-erythromycine-cephalexin (11.4%) . Strepto . pyogen was found resistant to tetracycline in 12.2% cases and to SMX-TMP in 8.3% cases.

Pol Merkuriusz Lek, 2003 Jan, 14(79), 5 - 8
{Alexander Project--5 years in Poland}; Hryniewicz W; The aim of the Alexander Project is to monitor resistance and its trends among the most common bacterial pathogens responsible for community-acquired lower respiratory tract infections and to compare the results obtained between different countries . In Poland, from 1996 to 2000 the percentage of Streptococcus pneumoniae isolates non-susceptible to penicillin was 14.1% . Most often decreased susceptibility to penicillin co-existed with resistance to other classes of antibiotics (multi-drug resistance) . The biggest number of pneumococci were resistant to co-trimoxazole (35.2%) and doxycycline (33.6%) . Of Haemophilus influenzae isolates, 5.3% produced beta-lactamases . All H . influenzae isolates were susceptible to amoxicillin/clavulanic acid and 3rd-generation cephalosporins . Many isolates of this species were resistant to co-trimoxazole (19.1%) . The results of the Alexander Project provide crucial data for guiding national recommendations for empirical treatment of respiratory tract infections.

Acta Paediatr, 2003, 92(2), 177 - 85
Parental perception of educational, behavioural and general health problems in school-age survivors of bacterial meningitis; Koomen I et al.; AIM: To determine the occurrence of educational, behavioural and general health problems in Dutch school-age survivors of bacterial meningitis . METHODS: A cohort of 680 school-age survivors of meningitis caused by the most common Gram-positive and Gram-negative bacteria was established approximately 6y after the children's illness . Children with Haemophilus influenzae type b (Hib) meningitis were excluded because this form of the disease has virtually disappeared . Parents completed questionnaires on educational, behavioural and general health problems . The reference group comprised 304 school-age siblings and peers . RESULTS: Postmeningitic children were more likely than controls to under achieve at school: 20% vs 5% (odds ratio 5.6; 95% confidence interval 3.0-10.7) . The postmeningitic children repeated a year twice as often as the children in the reference group (16% vs 8%, odds ratio: 2.5, 95% confidence interval 1.5-4.2) and were referred to a special-needs school four times more frequently (8% vs 2%, odds ratio: 5.5; 95% confidence interval 2.0-15.4) . Parents also reported more behavioural problems at home . More than half of the postmeningitic children experienced general health problems . The causative pathogen or age at infection had no influence on the relative frequency of educational and behavioural problems, and reduced auditory functioning played only a small part in these problems . CONCLUSION: Parents perceive educational, behavioural and general health problems in more than 30% of postmeningitic children . Until it is clear which children are at highest risk of developing these problems, it will be necessary to follow postmeningitic children into their school-age years.

Eur Arch Otorhinolaryngol, 2003 Apr, 260(4), 186 - 94 Epub 2002 Oct 11.
Comparison of the efficacy and safety of faropenem daloxate and cefuroxime axetil for the treatment of acute bacterial maxillary sinusitis in adults; Siegert R et al.; In this multicentre, multinational, comparative, double-blind clinical trial, outpatients with both clinical signs and symptoms and radiographic evidence of acute sinusitis were randomly assigned to receive for 7 days either a twice-daily oral regimen of faropenem daloxate (300 mg) or a twice daily oral regimen of cefuroxime axetil (250 mg) . Among 452 patients considered valid for clinical efficacy, faropenem daloxate treatment was found to be statistically equivalent to cefuroxime axetil (89.0% vs . 88.4%-95% CI=-5.2%; +6.4%) at the 7-16 days post-therapy assessment . At 28-35 days post-therapy, the continued clinical cure rate in the faropenem daloxate group was 92.6% and that for the cefuroxime axetil group was 94.9% (95% CI: -6.8%; +1.2%) . A total of 148 organisms was obtained in 136 microbiologically valid patients (30.1%) . The predominant causative organisms were Streptococcus pneumoniae (47.1%), Haemophilus influenzae (30.1%), Staphylococcus aureus (14.7%) and Moraxella catarrhalis (8.8%) . The bacteriological success rate at the 7-16 days post-therapy evaluation was similar in both treatment groups: 91.5% and 90.8% in the faropenem daloxate and cefuroxime axetil groups, respectively (95% CI=-9.2%; +9.5%) . Eradication or presumed eradication was detected for 97.3% and 96.3% of S . pneumoniae, 85.0% and 90.5% of H . influenzae, 88.9% and 90.9% of S . aureus and 100.0% and 83.3% of M . catarrhalis in faropenem daloxate and cefuroxime axetil recipients, respectively . At least one drug-related event was reported by 9.5% of the faropenem daloxate-treated patients and by 10.3% of those who received cefuroxime axetil . The most frequently reported drug-related events were diarrhoea (2.2% versus 2.9%), nausea/vomiting (1.5% vs . 0.7%), abdominal pain (0.7% vs 1.5%) and skin reactions (1.5% vs . 1.1%) . Overall, faropenem daloxate was at least as effective clinically and bacteriologically as cefuroxime axetil and was well tolerated.

Ambul Pediatr, 2003 May-Jun, 3(3), 154 - 7
Do laws bring children in child care centers up to date for immunizations?
Kolasa MS, Chilkatowsky AP, Stevenson JM, Lutz JP, Watson BM, Levenson R, Rosenthal J.
BACKGROUND: Pennsylvania state law requires licensed child care centers (CCCs) to document that each enrolled child is up to date (UTD) for routine immunizations within 60 days of enrollment . This study evaluates the law's impact on immunization coverage among children aged <or=59 months who attend CCCs in Philadelphia . METHODS: Out of Philadelphia's 440 commercial CCCs, 75 were randomly selected . Of these, 9 had closed, 3 did not accept children aged <or=59 months, and 3 refused assessment . For the remaining 60 CCCs, vaccination dates were abstracted from CCC records for all enrolled children <or=59 months . For children not UTD for all vaccines according to CCC records, additional data were sought from Philadelphia's immunization registry, health care providers, and parents . RESULTS: Records of 2847 children were assessed . According to CCC records, information from the immunization registry, vaccination providers, and parents, 71% of children aged 0-18 months, 77% of children 19-35 months, and 84% of children 36-59 months were UTD for their age for diphtheria, tetanus toxoids, and pertussis vaccine; polio; Haemophilus influenzae type b; and measles, mumps, and rubella vaccines . No significant increase in immunization coverage levels was found between the date children enrolled in a CCC and 60 days later . CONCLUSIONS: Up to one quarter of children <5 years of age enrolled in Philadelphia's CCCs are not UTD for immunizations, with children 0-18 months of age being most behind in their immunizations . Furthermore, many children do not receive vaccines within 60 days of enrollment . These low coverage levels combined with the potential exposures inherent in group care settings indicate that children in CCCs are at risk for contracting vaccine-preventable diseases.

Vaccine, 2003 May 16, 21(17-18), 2169 - 77
Analyzing the economic value of the hepatitis B--Haemophilus influenzae type B combination vaccine by reverse engineering a formulary selection algorithm; Jacobson SH et al.; Combination vaccines for pediatric immunization provide a means to reduce the number of separate injections required to immunize children . This paper reports the results of reverse engineering a vaccine selection algorithm to evaluate the economic value of a hepatitis B-Haemophilus influenzae type B combination vaccine that is currently under federal contract in the United States . This analysis captures the tradeoff between the cost assigned to administering an injection and the price of the vaccine that earns it a place in the lowest overall cost formulary . Given the current United States federally negotiated price for this combination vaccine (as of 9 August 2002), it provides a good economic value for those health-care providers or payers who value the cost associated with administering an injection to be at least US$ 4.02 or 5.01 . These two values are a function of the number of doses of the combination vaccine required to be in the lowest overall cost formulary and whether the perinatal hepatitis B dose is administered . Moreover, as the cost of an injection increases, the combination vaccine provides a better value than monovalent vaccines.

Vaccine, 2003 May 16, 21(17-18), 2107 - 14
Phase I study of a lipooligosaccharide-based conjugate vaccine against nontypeable Haemophilus influenzae; Gu XX et al.; Nontypeable Haemophilus influenzae (NTHi) accounts for about one-third of purulent otitis media (OM) in children and is a common cause of pulmonary infection in adults with decreased resistance . Based upon sero-epidemiological data in humans and immunochemical data in laboratory animals, a lipooligosaccharide (LOS)-tetanus toxoid (TT) conjugate was prepared and evaluated for its safety and immunogenicity in a Phase I study of 40 healthy adults . The conjugate was injected intramuscularly into all volunteers: 28 of them received a second injection 14 weeks later . Local and systemic reactions were monitored and sera, taken before and 2, 6, 14, 16, and 38 weeks after injection, were assayed for IgG, IgA, and IgM antibodies to the LOS by ELISA and for bactericidal activity . The results indicate that there were no significant local or systemic reactions after either injection . All volunteers had pre-existing IgG anti-LOS . The geometric mean (GM) level rose from 14 to 40 at 2 weeks, remained at 35 at 6 weeks (40 or 35 versus 14, P<0.01) and dropped to 27 at 14 weeks after the first injection . There was also a rise 2 weeks after the second injection (27 versus 37, P<0.05) . A total of 52.5% of subjects showed serum-conversion (greater than four-fold increase) after one and two injections . At 38 weeks, the GM IgG anti-LOS was still higher than before initial injection (20 versus 14, P<0.05) . A similar pattern of reactivity was observed for IgA and IgM anti-LOS . Similar to that observed in mice, but not in rabbits, the conjugate-induced antibodies did not yield significant bactericidal activity in vitro . The LOS-TT conjugate is well tolerant in adults and a Phase II evaluation of the conjugate in children is planned.

Vaccine, 2003 May 16, 21(17-18), 1907 - 12
Estimating the Haemophilus influenzae type b (Hib) disease burden and the impact of Hib vaccine in Fiji; Wilson N et al.; AIMS: To estimate Haemophilus influenzae type b (Hib) disease burden in Fiji in children under the age of 5 years (under-5s) prior to vaccine introduction . To compare estimates from WHO's Hib rapid assessment tool (RAT), with that from decline in disease after vaccine introduction . METHODS: Laboratory data (meningitis), hospitalization and mortality data (pneumonia and meningitis) before and after Hib vaccine introduction were collected . The RAT protocol provides two independent estimates of pre-vaccine disease burden (one based on meningitis incidence laboratory data and the other based on mortality statistics) . A third estimate uses the decline in disease following vaccine introduction . RESULTS: The decline in meningitis hospitalizations implies a pre-vaccine Hib meningitis incidence of 66 per 100,000 in under-5s . This compares with a pre-vaccine RAT estimate of Hib meningitis incidence of 84 per 100,000 (for 1992-1993) . The RAT estimated the total annual pre-vaccine Hib burden (meningitis plus pneumonia) at 476 cases and 36 deaths per year ("meningitis incidence method") and 70 cases and 5 deaths ("child mortality method") . Hib vaccine led to declines of 32% (95% confidence interval (CI)=11-48%), and 78% (95% CI=22-94%) for all under-5s meningitis hospitalizations and deaths, respectively . There was no similar consistent decline in pneumonia hospitalizations or deaths after vaccine introduction, except for a statistically significant reduction in pneumonia mortality in children aged under 1 year . CONCLUSIONS: Hib disease constitutes an important burden on the health of Pacific children that can be rapidly reduced with Hib vaccine . In this setting, routine morbidity statistics (comparing pre-and post-vaccine) provided an estimate of Hib meningitis burden which is broadly similar to that of the Hib RAT, suggesting that both might be valid ways to estimate Hib meningitis incidence . However, Hib pneumonia burden could not be estimated from routine statistics.

Vaccine, 2003 May 16, 21(17-18), 1894 - 900
Safety and immunogenicity of a heptavalent pneumococcal conjugate vaccine in infants; Zangwill KM et al.; OBJECTIVE: To evaluate the safety and immunogenicity of two lots of a heptavalent Streptococcus pneumoniae conjugate vaccine (PCV) containing seven capsular polysaccharide serotypes (4, 6B, 9V, 14, 18C, 19F, and 23F) conjugated to the outer membrane complex of Neisseria meningitidis serogroup B (OMPC) and administered to infants at 2, 4, 6, and 12 months of age . METHODS: One hundred twenty infants were randomly assigned to concurrently receive PCV-OMPC and one of two Haemophilus influenzae type b (Hib) conjugate-DTwP combination vaccines: (1) Hib with a heterologous protein carrier (CRM(197), TETRAMUNE, Group 1) or (2) an experimental Hib-hepatitis b combination vaccine with the homologous carrier (OMPC, Group 2) . All infants in Groups 1 and 2 received PCV-OMPC (lot 1) at 12 months of age . Another separate group of 120 infants (Group 3) received a different lot of PCV-OMPC concurrently with Hib-CRM(197) (TETRAMUNE) at 2, 4, and 6 months of age and then were randomized to receive either PCV-OMPC or a 23-valent polysaccharide (PS) pneumococcal vaccine at 12 months of age . RESULTS: Each PCV-OMPC lot was generally well tolerated and no vaccine-related serious adverse events were reported . Following the primary series, serotype-specific anti-pneumococcal geometric mean concentrations (GMC) were highest for serotypes 14, 19F, and 4 and lowest for serotypes 6B and 23F . GMC and seroconversion rates in Group 3 (lot 2) were lower than in Group 1 (lot 1) for serotypes 6B, 14, 18C, and 23F . Antibody responses to serotypes 6B, 14, and 18C were significantly lower in Group 2 compared to Group 1 . Following a booster dose of PCV-OMPC at 12 months of age, each lot was immunogenic with at least a 5-10-fold increase in antibody levels, and responses were significantly higher among those who received the PS vaccine . CONCLUSIONS: PCV-OMPC is generally safe in infants, displays variable immune response by serotype, and concomitant receipt of Hib vaccine with homologous carrier may impact on its immunogenicity.

Microbes Infect, 2003 Apr, 5(4), 275 - 83
Characterization of a locus encoding four paralogous outer membrane lipoproteins of Brachyspira hyodysenteriae; Cullen PA et al.; The identification of Brachyspira hyodysenteriae outer membrane proteins (OMPs) that may stimulate immunity to swine dysentery is important for vaccine development . We report here the analysis of a novel locus, blpGFEA, encoding four tandem paralogous proteins of approximately 30 kDa from B . hyodysenteriae . The four proteins share 31-39% sequence identity with lipoproteins from several species of bacterial pathogens, but the locus possesses a unique genetic organization . Using antisera raised to recombinant versions of each of these proteins, only BlpA and BlpE were found to be immunologically cross-reactive with the other proteins encoded by the locus . Northern hybridization indicated that only blpA was expressed under in vitro growth conditions . In addition, convalescent swine serum recognized recombinant BlpA in immunoblotting experiments, demonstrating that it is also expressed during infection . Analysis of the translated sequences of each of the genes revealed atypical spirochetal signal peptidase II recognition sites, and BlpA was shown to be a lipoprotein by incorporation of tritiated palmitic acid . Native BlpA was completely extracted by Triton X-114 (TX-114) and partitioned exclusively into the detergent phase during extraction of whole B . hyodysenteriae cells, implicating it as a component of the brachyspiral outer membrane . Consistent with the transcriptional and immunological data, analysis of the brachyspiral outer membrane proteome also revealed expression of only BlpA . Notably, inactivation of blpA homologs in Haemophilus influenzae and Salmonella enteritidis resulted in attenuation of virulence.

Clin Immunol, 2003 Mar, 106(3), 226 - 30
Infections with Haemophilus species in chronic granulomatous disease: insights into the interaction of bacterial catalase and H2O2 production; Kottilil S et al.; Chronic granulomatous disease (CGD) is a rare inherited disorder in which phagocytes are incapable of generating bactericidal-reactive oxygen derivatives . Typically these patients are susceptible to life-threatening infections with catalase-producing organisms . Haemophilus species, particularly H . paraphrophilus, are not associated with CGD infections, because these organisms rarely if ever produce catalase . Haemophilus species are part of the indigenous oral microbial flora and, other than H . influenzae, are rarely recognized as pathogens . They are fastidious and require additional growth factors and capnophilic culture conditions for optimal growth and identification . Here we describe three cases of infection with non-H . influenzae (NHI) Haemophilus species in CGD patients . These organisms were catalase-negative and therefore not expected to be virulent in CGD patients, but they were also H(2)O(2) production-negative, thereby negating the putative loss of virulence of being catalase-negative . These are the first reports of NHI Haemophilus species in CGD and reinforce the critical need for careful microbiologic evaluation of infections in CGD patients.

Microb Drug Resist, 2003 Spring, 9(1), 99 - 108
Risk factors for the nasopharyngeal carriage of respiratory pathogens by Portuguese children: phenotype and antimicrobial susceptibility of Haemophilus influenzae and Streptococcus pneumoniae; Neto AS et al.; Between 1997 and 2000 nasopharyngeal specimens were obtained from 466 children < or = 12 years old attending the Pediatric Emergency Department at S . Francisco Xavier Hospital, Lisbon, to evaluate risk factors for nasopharyngeal carriage of Haemophilus influenzae and Streptococcus pneumoniae and to characterize their phenotype and antimicrobial susceptibility . The attending pediatrician completed written questionnaires about the children's demographic and clinical histories . Over half the children (52.8%) carried H . influenzae and/or S . pneumoniae . Forty-one percent of these children had H . influenzae, 22.8% had S . pneumoniae and 36.2% had both . Risk factors identified for carriage of respiratory pathogens were: age below 3 years (p < 0.05), black race (p < 0.01), attending a daycare center (p < 0.05), and having a lower respiratory infection (p < 0.05) . Asthmatic children were less likely to be carriers (p = 0.004) . About two-thirds of H . influenzae isolates were susceptible to all antibiotics tested, 7.9% were beta-lactamase producers, 16.4% were nonsusceptible to trimethoprim, and 6.9% were intermediately resistant to clarithromycin . Over half (57.1%) of S . pneumoniae isolates were susceptible to all antibiotics tested, 21.1% were multiresistant, 23.3% were nonsusceptible to penicillin, and about 20% were resistant to macrolides . Low-level resistance to third-generation cephalosporins was detected in 2.3% . The data reflect the controversy surrounding risk factors of nasopharyngeal colonization . These may have significant implications on clinical practice and on antimicrobial strategies to prevent the appearance of further resistant strains . Our findings highlight the importance to investigate the relationship between asthma and carriage.

Microb Drug Resist, 2003 Spring, 9(1), 39 - 46
Diversity of ampicillin-resistance genes in Haemophilus influenzae in Japan and the United States; Hasegawa K et al.; Clinical isolates of Haemophilus influenzae from Japan (n = 296) and the United States (n = 100) were tested by the microdilution method for susceptibility in vitro to 10 beta-lactam antibiotics and molecular mechanisms of beta-lactam resistance . For all isolates, PCR was used to identify six elements, including beta-lactamase-producing ampicillin (AMP)-resistance (BLPAR) and beta-lactamase-nonproducing AMP-resistance (BLNAR) genes as follows: (1) TEM-1 type beta-lactamase gene, (2) ROB-1 type beta-lactamase gene, (3) part of normal ftsI gene encoding PBP3, which is involved in septal peptidoglycan synthesis, (4) a portion of the ftsI gene possessing some amino acid substitutions commonly detected in BLNAR strains, (5) p6 gene encoding P6 membrane proteins specific to H . influenzae, and (6) serotype b capsule gene . In Japanese and U.S . isolates, respective prevalences of each resistance class in Japan and the United States were 55.1% and 46% for beta-lactamase-nonproducing, AMP-susceptible (BLNAS); 3.0% and 26% for the TEM-1 type beta-lactamase gene; 0% and 10% for the ROB-1 type; 26.4% and 13% for low-BLNAR with a low degree of AMP resistance; and 13.2% and 0% for BLNAR strains . A few remaining isolates were beta-lactamase-producing strains with a mutation in the ftsI gene . MICs of all beta-lactam agents against low-BLNAR strains were 2-8 times higher than against BLNAS . MICs of cephalosporin antibiotics against BLNAR strains were 16-32 times higher than against BLNAS . The rank order of beta-lactam MIC90 values against BLNAR strains was piperacillin = ceftriaxone = cefditoren (0.25 microg/ml), meropenem (0.5), cefotaxime (1), AMP = cefpodoxime (8), cefdinir (16), amoxicillin (16), and cefaclor (64) . Serotype b isolates were few in both countries (2.4% in Japan, 3% in the United States) . Differences in proportions of respective AMP-resistant genes in H . influenzae isolates between the two countries might reflect differences in antibiotic agents ordinarily given to outpatients with community-acquired bacterial infections.

Infect Immun, 2003 May, 71(5), 2798 - 809
Role of surface-exposed loops of Haemophilus influenzae protein P2 in the mitogen-activated protein kinase cascade; Galdiero S et al.; The outer membrane of gram-negative bacteria contains several proteins, and some of these proteins, the porins, have numerous biological functions in the interaction with the host; porins are involved in the activation of signal transduction pathways and, in particular, in the activation of the Raf/MEK1-MEK2/mitogen-activated protein kinase (MAPK) cascade . The P2 porin is the most abundant outer membrane protein of Haemophilus influenzae type b . A three-dimensional structural model for P2 was constructed based on the crystal structures of Klebsiella pneumoniae OmpK36 and Escherichia coli PhoE and OmpF . The protein was readily assembled into the beta-barrel fold characteristic of porins, despite the low sequence identity with the template proteins . The model provides information on the structural features of P2 and insights relevant for prediction of domains corresponding to surface-exposed loops, which could be involved in the activation of signal transduction pathways . To identify the role of surface-exposed loops, a set of synthetic peptides were synthesized according to the proposed model and were assayed for MEK1-MEK2/MAPK pathway activation . Our results show that synthetic peptides corresponding to surface loops of protein P2 are able to activate the MEK1-MEK2/MAPK pathways like the entire protein, while peptides modeled on internal beta strands are unable to induce significant phosphorylation of the MEK1-MEK2/MAPK pathways . In particular, the peptides corresponding to loops L5 (Lys206 to Gly219), L6B (Ser239 to Lys253), and L7 (Thr280 to Lys287) activate, as the whole protein, essentially JNK and p38.

Infect Immun, 2003 May, 71(5), 2563 - 70
Cleavage of the human immunoglobulin A1 (IgA1) hinge region by IgA1 proteases requires structures in the Fc region of IgA; Chintalacharuvu KR et al.; Secretory immunoglobulin A (IgA) protects the mucosal surfaces against inhaled and ingested pathogens . Many pathogenic bacteria produce IgA1 proteases that cleave in the hinge of IgA1, thus separating the Fab region from the Fc region and making IgA ineffective . Here, we show that Haemophilus influenzae type 1 and Neisseria gonorrhoeae type 2 IgA1 proteases cleave the IgA1 hinge in the context of the constant region of IgA1 or IgA2m(1) but not in the context of IgG2 . Both C(alpha)2 and C(alpha)3 but not C(alpha)1 are required for the cleavage of the IgA1 hinge by H . influenzae and N . gonorrhoeae proteases . While there was no difference in the cleavage kinetics between wild-type IgA1 and IgA1 containing only the first GalNAc residue of the O-linked glycans, the absence of N-linked glycans in the Fc increased the ability of the N . gonorrhoeae protease to cleave the IgA1 hinge . Taken together, these results suggest that, in addition to the IgA1 hinge, structures in the Fc region of IgA are required for the recognition and cleavage of IgA1 by the H . influenzae and N . gonorrhoeae proteases.

Infect Immun, 2003 May, 71(5), 2478 - 86
Mutations in the lspA1 and lspA2 genes of Haemophilus ducreyi affect the virulence of this pathogen in an animal model system; Ward CK et al.; Haemophilus ducreyi 35000HP contains two genes, lspA1 and lspA2, whose predicted protein products have molecular weights of 456,000 and 543,000, respectively (C . K . Ward, S . R . Lumbley, J . L . Latimer, L . D . Cope, and E . J . Hansen, J . Bacteriol . 180:6013-6022, 1998) . We have constructed three H . ducreyi 35000HP mutants containing antibiotic resistance cartridges in one or both of the lspA1 and lspA2 open reading frames . Western blot analysis using LspA1- and LspA2-specific monoclonal antibodies indicated that the wild-type parent strain 35000HP expressed LspA1 protein that was readily detectable in culture supernatant fluid together with a barely detectable amount of LspA2 protein . The lspA2 mutant 35000HP.2 expressed LspA1 protein that was detectable in culture supernatant fluid and no LspA2 protein . In contrast, the H . ducreyi lspA1 mutant 35000HP.1, which did not express the LspA1 protein, expressed a greater quantity of the LspA2 protein than did the wild-type parent strain . The lspA1 lspA2 double mutant 35000HP.12 expressed neither LspA1 nor LspA2 . The three mutant strains adhered to human foreskin fibroblasts and to a human keratinocyte cell line in vitro at a level that was not significantly different from that of the wild-type strain 35000HP . Lack of expression of the LspA1 protein by both the lspA1 mutant and the lspA1 lspA2 double mutant was associated with an increased tendency to autoagglutinate . When evaluated in the temperature-dependent rabbit model for chancroid, the lspA1 lspA2 double mutant was substantially less virulent than the wild-type strain 35000HP . The results of these studies indicated that H . ducreyi requires both the LspA1 and LspA2 proteins to be fully virulent in this animal model for experimental chancroid.

Infect Immun, 2003 May, 71(5), 2384 - 93
Aae, an autotransporter involved in adhesion of Actinobacillus actinomycetemcomitans to epithelial cells; Rose JE et al.; The periodontal pathogen Actinobacillus actinomycetemcomitans possesses myriad virulence factors, among them the ability to adhere to and invade epithelial cells . Recent advances in the molecular manipulation of this pathogen and the sequencing of strain HK 1651 have facilitated examination of the genetics of its interaction with epithelial cells . The related gram-negative organism, Haemophilus influenzae, possesses autotransporter adhesins . A search of the sequence database of strain HK 1651 revealed a homologue with similarity in the pore-forming domain to that of the H . influenzae autotransporter, Hap . A . actinomycetemcomitans mutants deficient in the homologue, Aae, showed reduced binding to epithelial cells . A method for making A . actinomycetemcomitans SUNY 465 transiently resistant to spectinomycin was used with conjugation to generate an isogenic aae mutant . An allelic replacement mutant was created in the naturally transformable A . actinomycetemcomitans strain ATCC 29523 . Lactoferrin, an important part of the innate host defense system, protects against bacterial infection by bactericidal and antiadhesion mechanisms . Lactoferrin in human milk removes or cleaves Hap and another autotransporter, an immunoglobulin A1 protease, from the surface of H . influenzae, thereby reducing their binding to epithelial cells . Human milk whey had similar effects on Aae from A . actinomycetemcomitans ATCC 29523 and its binding to epithelial cells; however, there was little effect on the binding of SUNY 465 . A difference in the genetic structure of aae in the two strains, apparently due to the copy number of a 135-base repeated sequence, may be the cause of the differential action of lactoferrin . aae is the first A . actinomycetemcomitans gene involved in adhesion to epithelial cells to be identified.

Tunis Med, 2002 Aug, 80(8), 469 - 72
{Epidemiologic profile of Haemophilus influenzae infection in Tunisia}; Thabet L et al.; Haemophilus influenzae, a commensal bacteria, is frequently incriminated in broncho--pulmonary surinfections and severe diseases as meningitis, pneumonia and septic arthritis, particularly in young children . A multicenter study was conducted to establish the epidemiological profile of Haemophilus influenzae diseases, to determine the rate of antibiotics resistance for guide therapeutic and preventive strategies . The identification was based on the requirements for X and V factors, and the serotype b determined by agglutination . The betalactamase production was done by nitrocefin test . Antimicrobial susceptibility testing was determined on Muller Hinton chocolate agar with isovitalex . During the two year period, (January 1998 December 1999), 192 isolates of H . Influenzae were collected, 61% were recovered from invasive infections (44 meningitis, 8 bacteremia, 2 arthritis) . The serotype b was identified in 55.7% of cases, 67.3% were invasive strains . 24.5% of isolates were producing betalactamase particularly invasive serotype b strains . All isolates of H . influenzae were susceptible to cefotaxim and to ofloxacin . Resistance rates to other antibiotics were: erythromycin 56.2%, tetracyclin 10.3%, rifampin 12%, chloramphenicol 1%, cotrimoxazole 16.5%, 11.5% amikacin and 20% gentamicin . The incidence of meningitis remained frequent in our country, involving the introduction of the vaccination in official calendar . Nevertheless, the surveillance of H . influenzae invasives infections and the serotyping of isolates were necessary to evaluate the impact of the immunization.

J Clin Microbiol, 2003 Apr, 41(4), 1664 - 72
Haemophilus influenzae carriage in children attending French day care centers: a molecular epidemiological study; Dabernat H et al.; The nasopharyngeal Haemophilus influenzae flora of healthy children under the age of 3 years attending day care centers in three distinct French geographic areas was analyzed by sampling during two periods, spring 1999 (May and June) and fall 1999 (November and December) . The average carrier rate among 1,683 children was 40.9% . The prevalence of capsulated H . influenzae carriers was 0.4% for type f and 0.6% for type e . No type b strains were found among these children, of whom 98.5% had received one or more doses of anti-Haemophilus b vaccine . Among the strains, 44.5% were TEM-type beta-lactamase producers and nine (1.3%) were beta-lactamase-negative ampicillin-resistant strains . Pulsed-field gel electrophoresis restriction patterns showed a large diversity with 366 SmaI patterns from 663 strains . Among the strains isolated during a given period, 33% were isolated simultaneously in more than one area . In each area, depending on the sampling period, 68 to 72% of the strains had new pulsotypes and persistence of 28 to 32% of the strains was noted . For the 297 beta-lactamase-producing strains, 194 patterns were found . The genomic diversity of these strains was comparable to that of the whole set of strains and does not suggest a clonal diffusion . Among the beta-lactamase-producing strains isolated in November and December, depending on the area, 66 to 73% had new pulsotypes with persistence of only 27 to 33% of the strains . In any given geographic area, colonization by H . influenzae appears to be a dynamic process involving a high degree of genomic heterogeneity among the noncapsulated colonizing strains.

J Clin Microbiol, 2003 Apr, 41(4), 1623 - 36
Characterization of encapsulated and noncapsulated Haemophilus influenzae and determination of phylogenetic relationships by multilocus sequence typing; Meats E et al.; A multilocus sequence typing (MLST) scheme has been developed for the unambiguous characterization of encapsulated and noncapsulated Haemophilus influenzae isolates . The sequences of internal fragments of seven housekeeping genes were determined for 131 isolates, comprising a diverse set of 104 serotype a, b, c, d, e, and f isolates and 27 noncapsulated isolates . Many of the encapsulated isolates had previously been characterized by multilocus enzyme electrophoresis (MLEE), and the validity of the MLST scheme was established by the very similar clustering of isolates obtained by these methods . Isolates of serotypes c, d, e, and f formed monophyletic groups on a dendrogram constructed from the differences in the allelic profiles of the isolates, whereas there were highly divergent lineages of both serotype a and b isolates . Noncapsulated isolates were distinct from encapsulated isolates and, with one exception, were within two highly divergent clusters . The relationships between the major lineages of encapsulated H . influenzae inferred from MLEE data could not be discerned on a dendrogram constructed from differences in the allelic profiles, but were apparent on a tree reconstructed from the concatenated nucleotide sequences . Recombination has not therefore completely eliminated phylogenetic signal, and in support of this, for encapsulated isolates, there was significant congruence between many of the trees reconstructed from the sequences of the seven individual loci . Congruence was less apparent for noncapsulated isolates, suggesting that the impact of recombination is greater among noncapsulated than encapsulated isolates . The H . influenzae MLST scheme is available at it allows any isolate to be compared with those in the MLST database, and (for encapsulated isolates) it assigns isolates to their phylogenetic lineage, via the Internet.

J Biol Chem, 2003 Jul 4, 278(27), 24269 - 76 Epub 2003 Apr 14.
Porin OmpP2 of Haemophilus influenzae shows specificity for nicotinamide-derived nucleotide substrates; Andersen C et al.; Haemophilus influenzae has an absolute requirement for NAD (factor V) because it lacks all biosynthetic enzymes necessary for de novo synthesis of that cofactor . Therefore, growth in vitro requires the presence of NAD itself, NMN, or nicotinamide riboside (NR) . To address uptake abilities of these compounds, we investigated outer membrane proteins . By analyzing ompP2 knockout mutants, we found that NAD and NMN uptake was prevented, whereas NR uptake was not . Through investigation of the properties of purified OmpP2 in artificial lipid membrane systems, the substrate specificity of OmpP2 for NAD and NMN was determined, with KS values of approximately 8 and 4mm, respectively, in 0.1 m KCl, whereas no interaction was detected for the nucleoside NR and other purine or pyrimidine nucleotide or nucleoside species . Based on our analysis, we assume that an intrinsic binding site within OmpP2 exists that facilitates diffusion of these compounds across the outer membrane, recognizing carbonyl and exposed phosphate groups . Because OmpP2 was formerly described as a general diffusion porin, an additional property of acting as a facilitator for nicotinamide-based nucleotide transport may have evolved to support and optimize utilization of the essential cofactor sources NAD and NMN in H . influenzae.

Arch Pediatr Adolesc Med, 2003 Apr, 157(4), 389 - 92
Risk of bacteremia in young children with pneumonia treated as outpatients; Shah SS et al.; BACKGROUND: Blood cultures are often obtained as part of the evaluation of children with pneumonia . There are few data regarding the risk of bacteremia with pneumonia in children since introduction of the Haemophilus influenzae type b vaccine . OBJECTIVE: To evaluate the risk of bacteremia in young children with pneumonia who were treated as outpatients . METHODS: A retrospective cohort study of 580 children aged 2 to 24 months who were evaluated by blood culture in a tertiary care children's hospital emergency department between February 1, 1993, and May 31, 1996, and discharged with the diagnosis of pneumonia . RESULTS: The mean patient age was 14.1 months; 339 patients (58.4%) were boys . Thirty-eight patients (6.6%) reported the use of oral antibiotics before initial emergency department evaluation . The prevalence of bacteremia was 1.6% (95% confidence interval, 0.7%-2.9%) . Streptococcus pneumoniae was the causative organism in all 9 cases . The serotype was available for 8 of 9 cases . Six (75%) of 8 cases of S pneumoniae bacteremia were caused by serotypes included in the current heptavalent pneumococcal conjugate vaccine, which was not available at the time of this study . The contamination rate was 1.9% (95% confidence interval, 1.0%-3.4%) . The mean +/- SD time to blood culture positive for organisms in a continuously monitored system was significantly shorter for pathogens (13.9 +/- 1.3 hours) than for contaminants (21.2 +/- 6.1 hours; P =.01) . CONCLUSIONS: Children aged 2 to 24 months with pneumonia who are treated as outpatients are at low risk of bacteremia . Widespread use of the pneumococcal conjugate vaccine may further decrease the incidence of bacteremia in this population.

Mol Microbiol, 2003 May, 48(3), 737 - 51
The Haemophilus influenzae HMW1 adhesin is glycosylated in a process that requires HMW1C and phosphoglucomutase, an enzyme involved in lipooligosaccharide biosynthesis; Grass S et al.; Non-typeable Haemophilus influenzae is a common respiratory pathogen and an important cause of morbidity in humans . The non-typeable H . influenzae HMW1 and HMW2 adhesins are related proteins that mediate attachment to human epithelial cells, an essential step in the pathogenesis of disease . Secretion of these adhesins requires accessory proteins called HMW1B/HMW2B and HMW1C/HMW2C . In the present study, we investigated the specific function of HMW1C . Examination of mutant constructs demonstrated that HMW1C influences both the size and the secretion of HMW1 . Co-immunoprecipitation and yeast two-hybrid assays revealed that HMW1C interacts with HMW1 and forms a complex in the cytoplasm . Additional experiments and homology analysis established that HMW1C is required for glycosylation of HMW1 and may have glycotransferase activity . The glycan structure contains galactose, glucose and mannose and appears to be generated in part by phosphoglucomutase, an enzyme important for lipooligosaccharide biosynthesis . In the absence of glycosylation, HMW1 is partially degraded and is efficiently released from the surface of the organism, resulting in reduced adherence . Based on these results, we conclude that glycosylation is a prerequisite for HMW1 stability . In addition, glycosylation appears to be essential for optimal HMW1 tethering to the bacterial surface, which in turn is required for HMW1-mediated adherence, thus revealing a novel mechanism by which glycosylation influences cell-cell interactions.

Biochemistry, 2003 Apr 22, 42(15), 4463 - 75
Structure of extended lipopolysaccharide glycoforms containing two globotriose units in Haemophilus influenzae serotype b strain RM7004; Masoud H et al.; Lipopolysaccharide (LPS) is a major virulence determinant of the human bacterial pathogen Haemophilus influenzae . Structural elucidation of the LPS from H . influenzae type b strain RM7004 was achieved by using electrospray ionization mass spectrometry (ESI-MS) and high-field NMR techniques on delipidated LPS and core oligosaccharide samples of LPS . It was found that the organism elaborates a series of related LPS glycoforms having a common inner-core structure, but differing in the number and position of attached hexose residues . LPS glycoforms containing between four and nine hexose residues were structurally characterized . The inner-core element was determined to be L-alpha-D-Hepp-(1-->2)-{PEA-->6}-L-alpha-D-Hepp-(1-->3)-{beta-D-Glcp-(1-->4)}-L-alpha-D-Hepp-(1-->5)-{P-->4}-alpha-KDOp-(2-->, a structural feature which has been identified in every H . influenzae strain investigated to date . Two major groups of isomeric glycoforms were characterized in which the terminal Hepp residue of the inner-core element was either substituted at the O-2 position with a beta-D-Galp residue or not . The structures of the major LPS glycoforms were found to have oligosaccharide chain extensions from O-3 of the middle Hepp residue . Glycoforms containing five and six hexose residues were most abundant and were shown to carry the tetrasaccharide unit alpha-D-Galp-(1-->4)-beta-D-Galp-(1-->4)-beta-D-Glcp-(1-->4)-alpha-D-Glcp at the O-3 position of the middle heptose . This tetrasaccharide displays the globoside trisaccharide (globotriose) as a terminal epitope, a structure that is found on many human cells (P(k) blood group antigen) and which is thought to be an important virulence determinant for H . influenzae . LPS glycoforms were characterized that had further chain extension from the beta-D-Glcp-(1--> residue of the proximal Hepp . In the fully extended LPS (Hex9/Hex8' glycoforms), both the proximal and middle heptose residues carried tetrasaccharide chains displaying terminal globotriose epitopes . In addition, the LPS was found to carry phosphorylcholine and O-acetyl groups.

Respir Med, 2003 Apr, 97(4), 375 - 81
Haemophilus parainfluenzae infection of respiratory mucosa; Middleton AM et al.; The pathogenicity of Haemophilus parainfluenzae (Hpi) in the respiratory tract is unclear, in contrast to the accepted pathogenicity of its close relative non-typable H . influenzae . We have investigated the interaction of two Hpi isolates with the mucosa of adenoid and bronchial tissue organ cultures . The adherence of bacteria to the mucosa of organ cultures, the effect of broth culture filtrates on human nasal epithelium, and interleukin (IL)-8 production by A549 cell cultures was investigated . Hpi 4846 adhered infrequently in clusters of pleomorphic cocco-bacilli to areas of epithelial damage, mucus and unciliated cells in adenoid organ culture experiments at 24 h, but not bronchial mucosa . Hpi 3698 was seen in only one adenoid and no bronchial organ cultures at 24 h . In separate experiments, Hpi 3698 was cleared more rapidly from the centre of the adenoid organ culture and was not cultured at 24 h . Although not adhering to the mucosa at 24 h, Hpi 3698, but not Hpi 4846, caused an increase in the amount of epithelial damage in both types of organ culture . Broth culture filtrates of both strains caused immediate slowing of ciliary beat frequency that progressed, and disrupted epithelial integrity . Dialysed culture filtrates of both strains stimulated IL-8 production by A549 cells, with the culture filtrate of Hpi 3698 being most potent . We conclude that two strains of Hpi varied in their adherence to adenoid tissue, and neither adhered to bronchial tissue . These results lead us to speculate that Hpi is only likely to be a pathogen in the lower respiratory tract when impaired airway defences delay bacterial clearance.

Am J Respir Cell Mol Biol, 2003 Nov, 29(5), 598 - 605 Epub 2003 Apr 14.
Nontypeable Haemophilus influenzae activates human eosinophils through beta-glucan receptors; Ahren IL et al.; Eosinophils are a characteristic component of the inflammatory response seen in several diseases, including allergic asthma and chronic obstructive pulmonary disease . After activation, eosinophil-derived products may exert proinflammatory effects and cause considerable tissue damage . In the present study, we investigated innate interactions between the respiratory tract pathogen nontypeable Haemophilus influenzae (NTHi) and human eosinophils . Bacterial binding to eosinophils was dependent on (1-3)-beta-D-glucan receptors, as deduced from blocking experiments using the soluble glucan derivatives laminarin and scleroglucan . In addition, expression of the beta-glucan receptor dectin-1 was shown in eosinophils by reverse transcriptase-polymerase chain reaction . Activation of the beta-glucan receptors by bacteria elicited a time- and dose-dependent respiratory burst in eosinophils . NTHi caused increased expression of the proinflammatory chemokine interleukin-8 as measured by reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay . Incubation of eosinophils in the presence of NTHi for 4.5 h revealed upregulation of 245 different genes as detected by microarray . Signal transduction-related transcripts were most strongly upregulated, followed by cytokine mRNAs . Our findings suggest that NTHi can induce an innate inflammatory response in eosinophils that is mainly mediated via beta-glucan receptors . This points to possible pathophysiologic mechanisms involving innate recognition of NTHi by eosinophils during infection of the airways, thus promoting inflammation in chronic pulmonary disease.

Curr Opin Pulm Med, 2003 May, 9(3), 227 - 32
Vaccine development for capsulate bacteria causing pneumonia; Russell FM et al.; Pneumonia strikes the extremes of the age spectrum, causing maximal death and disability in children and the elderly . Despite its worldwide impact, there is a paucity of epidemiologic data regarding its incidence and the causative organisms . The two leading causes of bacterial pneumonia in childhood are Streptococcus pneumoniae (SP) and Haemophilus influenzae type b (Hib) . SP is the major cause of pneumonia beyond the newborn period . In neonates, Group B Streptococcus (GBS) remains a major cause of sepsis and pneumonia despite recent reductions due to targeted perinatal antibiotic prophylaxis . Hib vaccine can prevent pneumonia in developing countries . SP conjugate vaccine prevents X-ray confirmed pneumonia in low incident populations, but protection appears more marginal in high incident populations . Non-vaccine SP serotypes have demonstrated increased carriage and mucosal disease, but not invasive disease following vaccination . GBS vaccines are in the early stages of clinical development as prenatal or antenatal vaccines.

Curr Opin Pulm Med, 2003 May, 9(3), 171 - 4
Acute and chronic frontal sinusitis; Brook I; This review summarizes the recent literature published on the microbiology, diagnosis, and medical and surgical management of acute and chronic frontal sinus disease . Two retrospective studies investigated the microbiology of frontal sinusitis in patients that underwent sinus surgery . Streptococcus pneumoniae, Haemophilus influenzae, and Staphylococcus aureus predominated in acute infection in patients with acute frontal sinusitis, and S . aureus and anaerobic bacteria were commonly isolated in chronic sinusitis . Surgery is indicated to treat patients with acute and chronic sinusitis and their complications . Several surgical procedures were recently evaluated, and these are briefly reviewed.

J Immunol, 2003 Apr 15, 170(8), 4373 - 9
Development of peptide mimotopes of lipooligosaccharide from nontypeable Haemophilus influenzae as vaccine candidates; Hou Y et al.; Nontypeable Haemophilus influenzae (NTHi) is a common cause of otitis media in children and lower respiratory tract diseases in adults . So far there is no effective vaccine against NTHi . A major surface-exposed component of NTHi, lipooligosaccharide (LOS), is a virulence factor as well as a potential protective Ag . LOS is too toxic to be administered in humans . However, detoxified LOS is a T cell-independent small molecule and is poorly immunogenic in vivo, so we converted LOS into a nontoxic T cell-dependent Ag through the use of peptides that mimic the LOS by screening a phage-display peptide library with a rabbit Ab specific for NTHi LOS . Fifty-six phage clones were found to share LOS mimicry molecules . Among them, 22 clones were subjected to DNA sequencing, and four consensus sequences were identified as NMMRFTSQPPNN, NMMNYIMDPRTH, NMMKYISPPIFL, and NMMRFTELSTPS . Three of the four synthetic peptides showed strong binding reactivity to the rabbit anti-LOS Ab and also a mouse bactericidal monoclonal anti-LOS Ab in vitro, and elicited specific serum anti-LOS Abs in rabbits (27- to 81-fold) after conjugation with keyhole limpet hemocyanin . Passive immunization with the rabbit antisera resulted in a significantly enhanced pulmonary bacterial clearance in a mouse model . The enhanced bacterial clearance was eliminated if the rabbit serum was preabsorbed with NTHi LOS . These data indicate that the peptide mimotopes of LOS that we have identified might be potential components of peptide vaccines against NTHi.

Eur Arch Otorhinolaryngol, 2003 Mar, 260(3), 141 - 7 Epub 2002 Oct 03.
Mucin in middle ear effusions inhibits attachment of Haemophilus influenzae to mucosal epithelial cells; Solzbacher D et al.; Although otitis media with effusion is often preceded by an infection of the tympanic cavity, when cultured, many effusions show no culturable bacteria . Based on the hypothesis that the effusion might play a protective role in the course of infection, the influence of this fluid on adhesion of H . influenzae (Hi) type-b strain 770235 and nontypeable H . influenzae (NTHi) strains to buccal epithelial cells was investigated . Effusions were classified as mucoid, seromucoid and serous . Mucoid secretions inhibited adhesion to a significantly greater extent (62%) than did seromucous (52%) and serous effusions (47%) ( P<0.001) . The glycoprotein and high-molecular-weight fractions showed similar levels of inhibition . Sialic acid concentration, and, to a lesser extent, protein concentration, correlated with the level of inhibition . Desialylated effusions lost their ability to block bacterial attachment . Thus, middle ear effusion fluid exhibits an inhibitory effect that is due to mucins, which determine viscosity and represent the sialylated high-molecular-weight glycoprotein fraction of the effusion.

J Pharmacol Sci, 2003 Jan, 91(1), 1 - 7
Exploitation of host epithelial signaling networks by respiratory bacterial pathogens; Li JD; Although tremendous effort has been put towards identifying the surface molecules of nontypeable Haemophilus influenzae (NTHi) for vaccine development over the past decades, it is only recently that we have begun to appreciate the intricate host epithelial signaling networks activated by NTHi, an important human pathogen causing respiratory infections . From what has been reported, it is evident that NTHi activates multiple signaling pathways in host epithelial cells that, in turn, inadvertently contribute to the pathogenesis . Among those signaling pathways, activation of NF-kappaB leads to up-regulation of IL-1beta, IL-8 and TNF-alpha, mucin MUC2 and Toll-like receptor 2 (TLR2), whereas activation of p38 MAP kinase mediates not only up-regulation of inflammatory mediators and mucin MUC5AC but also down-regulation of TLR2 . Interestingly, NTHi-induced activation of the PI3K-Akt pathway, however, leads to inhibition of p38 mitogen-activated protein (MAP) kinase . Moreover, the TGF-beta-Smad signaling pathway cooperates with NF-kappaB to mediate up-regulation of mucin MUC2 . Finally, glucocorticoids synergistically enhance NTHi-induced TLR2 expression via specific up-regulation of the MAP kinase phosphatase-1 that, in turn, leads to inactivation of p38 MAP kinase, the negative regulator for TLR2 expression . These studies may bring new insights into the molecular pathogenesis of NTHi-induced infections and open up novel therapeutic targets for these diseases.

Scand J Infect Dis, 2003, 35(1), 21 - 6
Resistance in respiratory tract pathogens and antimicrobial use in Icelandic and Lithuanian children; Hjaltested EK et al.; To assess resistance rates among respiratory tract pathogens and antimicrobial usage of children in Iceland and Lithuania, and to correlate usage patterns with resistance rates, nasopharyngeal swabs were taken from healthy children attending day-care centres in Reykjavik (n = 6) and Vilnius (n = 13), and cultured for pneumococci, Haemophilus influenzae and Moraxella catarrhalis . Information about recent antimicrobial use was obtained by a questionnaire . 50% and 51% carried pneumococci, 59% and 67% carried H . influenzae and 60% and 46% carried M . catarrhalis, in Reykjavik (n = 297) and Vilnius (n = 508), respectively . The prevalence of penicillin non-susceptible pneumococci was 11% in Reykjavik (mainly serotype 6B) and 5% in Vilnius (mainly 23F) . At the time of sampling 3% of the children in Reykjavik and 8% in Vilnius were receiving antimicrobials . Children carrying penicillin non-susceptible pneumococci had used antimicrobials more frequently in the preceding month and this was significant for the Icelandic children (p < 0.0002) . Antimicrobial use was more common in Vilnius than in Reykjavik, yet antimicrobial resistance was more common in Reykjavik . Resistance rates cannot be predicted only on the basis of recent antimicrobial use.

J Med Microbiol, 2003 Apr, 52(Pt 4), 277 - 82
Haemophilus influenzae Rd KW20 has virulence properties; Daines DA et al.; Haemophilus influenzae is a human-adapted commensal and pathogen that can cause mucosal infections such as sinusitis, otitis media and bronchitis . Certain strains also cause bacteraemia and meningitis . Clinical isolates are genetically heterogeneous and are often recalcitrant to standard genetic manipulation . H . influenzae strain Rd KW20 has traditionally been considered avirulent, since it does not survive in the bloodstream of animals, is readily killed by normal adult human sera and cannot colonize the nasopharynx of infant rats . The purpose of this study was to determine whether Rd KW20 could be used in certain infection models . It is shown here that strain Rd KW20 can invade certain human epithelial cell lines grown either as monolayers or as differentiated epithelium at the air-liquid interface . In addition, Rd KW20 can invade a monolayer of immortalized human brain microvascular endothelial cells . Finally, this strain can replicate and survive in human bronchial xenografts for up to 3 weeks . The complete genomic sequence of Rd KW20 is available and it is readily amenable to genetic manipulation . These properties and the results reported here indicate that this strain is a viable alternative to the use of clinical isolates for the investigation of H . influenzae virulence.

Mol Microbiol, 2003 Apr, 48(2), 323 - 34
Neisseria meningitidis App, a new adhesin with autocatalytic serine protease activity; Serruto D et al.; Neisseria meningitidis is a Gram-negative bacterium which colonizes the human upper respiratory tract . Occasionally, it translocates to the bloodstream causing sepsis and from there it can cross the blood-brain barrier and cause meningitis . Many of the molecules, which mediate the interaction of N . meningitidis to host cells, are still unknown . Recently, App (Adhesion and penetration protein) was described as a member of the autotransporter family and a homologue to the Hap (Haemophilus adhesion and penetration) protein of Haemophilus influenzae, a molecule that plays a role in the interaction with human epithelial cells . In this study we expressed app in Escherichia coli in order to analyse the functional properties of the protein . We show that the protein is exported to the E . coli surface, processed by an endogenous serine-protease activity and released in the culture supernatant . Escherichia coli expressing app adhere to Chang epithelial cells, showing that App is able to mediate bacterial adhesion to host cells . The serine protease activity is localized at the amino-terminal domain, whereas the binding domain is in the carboxy-terminal region . The role of App in adhesion was confirmed also in N . meningitidis.

J Infect Chemother, 2003 Mar, 9(1), 46 - 52
Current status of bacterial resistance in the otolaryngology field: results from the Second Nationwide Survey in Japan; Suzuki K et al.; The study reported here was a nationwide assessment of otitis media (466 patients with acute suppurative otitis media and 476 with chronic suppurative otitis media), sinusitis (447 with acute sinusitis and 426 with chronic sinusitis), acute tonsillitis (724 patients), and peritonsillar abscess (141 patients) performed between November 1998 and March 1999 . Eighty university hospitals, 79 affiliated hospitals, and 103 general practitioners participated . Methicillin-resistant Staphylococcus aureus(MRSA) comprised 15.6% of the 786 isolated strains of S . aureus . MRSA was frequently detected in patients with suppurative otitis media, but was uncommon in those with acute tonsillitis or peritonsillar abscess, and it was more common in those who had already been treated than in those who had not, with a significant difference between the groups . Vancomycin (VCM) showed the highest antimicrobial activity against MRSA and no VCM resistance was detected . Penicillin-sensitive Streptococcus pneumoniae(PSSP), penicillin-intermediate-resistant S . pneumoniae (PISP), and penicillin-resistant S . pneumoniae (PRSP) accounted for 49.6%, 28.5%, and 21.9% of the 228 isolated strains of S . pneumoniae, respectively . PISP and PRSP were frequently detected in children aged 5 years or younger . beta-Lactamase was produced by 96 of the 100 strains (96%) of Moraxella (Branhamella) catarrhalis . The 281 strains of Haemophilus influenzae isolated consisted of 199 beta-lactamase-negative, ampicillin-sensitive (BLNASe) strains (70.8%), 65 beta-lactamase-negative ampicillin-resistant (BLNAR) strains (23.1%), and 17 beta-lactamase-producing strains (6.0%) . BLNAR strains were frequently detected in pretreated patients . Of these 281 strains of H . influenzae, 214 had nontypable capsules . In conclusion, the major bacterial species showed resistance to beta-lactams, indicating that care should be taken when selecting an appropriate antimicrobial agent.

J Bacteriol, 2003 Apr, 185(8), 2528 - 37
Porphyrin-mediated cell surface heme capture from hemoglobin by Porphyromonas gingivalis; Paramaesvaran M et al.; The porphyrin requirements for growth recovery of Porphyromonas gingivalis in heme-depleted cultures are investigated . In addition to physiologically relevant sources of heme, growth recovery is stimulated by a number of noniron porphyrins . These data demonstrate that, as for Haemophilus influenzae, reliance on captured iron and on exogenous porphyrin is manifest as an absolute growth requirement for heme . A number of outer membrane proteins including some gingipains contain the hemoglobin receptor (HA2) domain . In cell surface extracts, polypeptides derived from HA2-containing proteins predominated in hemoglobin binding . The in vitro porphyrin-binding properties of a recombinant HA2 domain were investigated and found to be iron independent . Porphyrins that differ from protoporphyrin IX in only the vinyl aspect of the tetrapyrrole ring show comparable effects in competing with hemoglobin for HA2 and facilitate growth recovery . For some porphyrins which differ from protoporphyrin IX at both propionic acid side chains, the modification is detrimental in both these assays . Correlations of porphyrin competition and growth recovery imply that the HA2 domain acts as a high-affinity hemophore at the cell surface to capture porphyrin from hemoglobin . While some proteins involved with heme capture bind directly to the iron center, the HA2 domain of P . gingivalis recognizes heme by a mechanism that is solely porphyrin mediated.

Int J Pediatr Otorhinolaryngol, 2003 Apr, 67(4), 317 - 21
Paediatric acute epiglottitis: not a disappearing entity; McEwan J et al.; OBJECTIVE: Paediatric epiglottitis is a serious, potentially life-threatening condition . Since the widespread introduction of the Haemophilus influenzae type b (Hib) conjugate vaccine in the UK in October 1992, there has been a dramatic reduction in its incidence . Vaccine failure is rare . The purpose of this study is to examine the failure rate of H . influenzae type b vaccine as measured by the number of cases of Haemophilus epiglottitis in fully vaccinated children presenting to a tertiary paediatric centre . A secondary aim is to provide a retrospective review of all cases of epiglottitis over a 13-year period . METHODS: A retrospective case-note review identifying all cases of epiglottitis presenting to Alder Hey Hospital was undertaken covering the time period December 1987-January 2001 . Details of patient age, sex, source of referral, clinical presentation, management and complications along with microbiological and serological findings were obtained . There were 21 males and 19 females . The mean age was 36 months (range 6-125 months) . A provisional diagnosis was made on the basis of the clinical features, confirmed by direct laryngoscopy in all but two cases and further supported in 28 cases by a positive blood culture . Of the 40 children presenting with epiglottitis, eight (20%) presented after the introduction of the Hib conjugate vaccine . H . influenzae antibody titres were measured both in the acute and convalescent phases of illness by the central Haemophilus Reference Unit in Oxford . RESULTS: We present the clinical features, management and complications of 40 cases of acute epiglottitis . H . influenzae was isolated from blood cultures in 28 cases (70%) . In 12 of these cases, H . influenzae type b was identified, seven prior to 1993 and five thereafter . Four of these five cases presenting after introduction of the Hib vaccine were known to have been fully vaccinated . One child had a history of prematurity and serum immunoglobulin estimation was abnormally low in another child . Acute Hib antibody titre was less than 1 microg/ml in two of the three cases in which this was available . CONCLUSION: Whilst the incidence of Haemophilus type b epiglottitis has significantly diminished, vaccine failure does occur . We discuss the current understanding of clinical and immunological risk factors for vaccine failure and the significance of the Hib antibody titre . Further evaluation of vaccine failure would be of benefit . The series that we present highlights the importance of considering acute epiglottitis in the differential diagnosis of the child presenting with acute upper airway obstruction . This is particularly relevant when in future there will be fewer doctors familiar with the symptoms and signs of the disease.

Clin Pediatr (Phila), 2003 Mar, 42(2), 93 - 9
Vaccine interchangeability; Greenberg DP et al.; The recommended childhood immunization schedule is complex, with multiple doses of vaccines required to complete the series . For a variety of reasons, a single manufacturer's product may not be available for all of the doses needed . In addition, combination products may become available, allowing for fewer injections to be administered but resulting in a change of vaccine components . To ensure the timely immunization of children and to avoid missed opportunities to vaccinate, it is important to be familiar with data documenting the interchangeability of vaccines . Although clear evidence is available to demonstrate the interchangeability of some vaccines (i.e., hepatitis B vaccines and Haemophilus influenzae type b vaccines), it is more difficult to evaluate the interchangeability of others (i.e., diphtheria-tetanus-acellular pertussis {DTaP} vaccines) . Limited data support the interchangeability of some DTaP products when necessary . This article presents the safety and immunogenicity data for immunization regimens that use vaccines from different manufacturers.

Mol Microbiol, 2003 Apr, 48(1), 117 - 29
A novel cell-binding mechanism of Moraxella catarrhalis ubiquitous surface protein UspA: specific targeting of the N-domain of carcinoembryonic antigen-related cell adhesion molecules by UspA1; Hill DJ et al.; Carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) are receptors for several Neisseria and Haemophilus spp . In this investigation, we demonstrate that a major outer membrane protein of Moraxella catarrhalis (Mx) strains, belonging to the ubiquitous surface protein (Usp) family, also interacts with the receptor . The interaction was demonstrated in Western blot overlay of SDS-PAGE-separated bacterial proteins using soluble receptor constructs as well as by co-precipitation experiments . The identity of the bacterial ligand was further ascertained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) . It was shown to belong to the UspA1 subfamily . In general, antibodies raised against synthetic UspA1, but not UspA2, peptides bound to the Mx ligand . CEACAM1-Fc-binding property could be demonstrated in all the clinical isolates examined but varied between strains . A single colony derivative of an Mx isolate was also demonstrated to bind to transfected Chinese hamster ovary and some human respiratory epithelial cells in a CEACAM-dependent manner . Thus, we have identified the third respiratory pathogen with the capacity to target the CEACAM family of receptors . The Mx ligand is structurally unrelated to those of Neisseria and Haemophilus.

Drugs, 2003, 63(7), 673 - 82; discussion 683-4
DTPa-HBV-IPV/Hib vaccine (Infanrix hexa); Curran MP et al.; Primary vaccination of infants with diphtheria-tetanus-acellular pertussis-hepatitis B recombinant (adsorbed)-inactivated poliomyelitis-adsorbed conjugated Haemophilus influenzae type b vaccine (DTPa-HBV-IPV/Hib; Infanrix hexa)-inactivated poliomyelitis-absorbed conjugated Haemophilus influenzae type b vaccine (DTPa-HBV-IPV/Hib) refers to Infanrix hexa trade mark.) provided high levels of seroprotection against diphtheria toxoid, tetanus toxoid, poliovirus 1, 2 and 3, pertussis antigens (pertussis toxoid, filamentous haemagglutinin and pertactin), hepatitis B virus surface antigen and H . influenzae polyribosyl-ribitol-phosphate (PRP) antigen . Most infants (97%) had anti-PRP levels >/=0.15 micro g/mL after a booster dose at 18 months . Primary vaccination with the DTPa-HBV-IPV/Hib vaccine produced a similar immune response to that with two different pentavalent plus monovalent vaccine combinations . Coadministration of DTPa-HBV-IPV/Hib vaccine and a heptavalent pneumonococcal conjugate vaccine resulted in a high level of seroprotection and was well tolerated . Primary or booster vaccination with DTPa- HBV-IPV/Hib vaccine was well tolerated . Commonly reported local adverse reactions included redness, pain and swelling . Systemic symptoms were usually mild to moderate, and included fussiness, fever, restlessness and sleepiness.

Infect Immun, 2003 Apr, 71(4), 1635 - 42
Prevalence and distribution of adhesins in invasive non-type b encapsulated Haemophilus influenzae; Rodriguez CA et al.; Adhesion to the respiratory epithelium plays an important role in Haemophilus influenzae infection . The distribution of H . influenzae adhesins in type b and nontypeable strains has been characterized, but little is known about the prevalence of these factors in non-type b encapsulated strains . We analyzed 53 invasive type a, type e, and type f strains for the presence of hap, hia, hmw, and hif genes; Hap, Hia, and HMW1/2 adhesins; and hemagglutinating pili . The hap gene was ubiquitous, and homologs of hmw and hia were present in 7 of 53 (13.2%) and 45 of 53 (84.9%) strains, respectively . Hap was detected in 28 of 45 (62.2%) hap(+) strains, HMW1/2 was detected in 5 of 7 (71.4%) hmw(+) strains, and Hia was detected in 31 of 45 (68.8%) hia(+) strains . The hif gene cluster was present in 26 of 53 strains (49.1%), and 21 of 26 hif(+) strains (80.8%) agglutinated (HA) red blood cells . Nine isolates exhibited HA but lacked the hif gene cluster . The distribution of adhesin genes correlated with the genetic relatedness of the strains . Strains belonging to one type a clonotype and the major type e clonotype possessed hia but lacked the hif cluster . Strains belonging to the second type a clonotype possessed both hia and hif genes . All type f strains belonging to the major type f clonotype possessed hia and lacked hifB . Although the specific complement of adhesin genes in non-type b encapsulated H . influenzae varies, most invasive strains express Hap and Hia, suggesting these adhesins may be especially important to the virulence of these organisms.

Antimicrob Agents Chemother, 2003 Apr, 47(4), 1308 - 12
Bactericidal activities of methoxyfluoroquinolones gatifloxacin and moxifloxacin against aerobic and anaerobic respiratory pathogens in serum; Stein GE et al.; Gatifloxacin (Bristol-Myers Squibb) and moxifloxacin (Bayer) are new methoxyfluoroquinolones with broad-spectrum activity against aerobic and anaerobic pathogens of the respiratory tract . In this investigation, we analyzed the bactericidal activity in serum over time of these antimicrobials against three aerobic (Streptococcus pneumoniae, Haemophilus influenzae, and Staphylococcus aureus) and four anaerobic (Peptostreptococcus micros, Peptostreptococcus magnus, Fusobacterium nucleatum, and Prevotella melaninogenica) bacteria associated with respiratory tract infections . Serum samples were obtained from 11 healthy male subjects following a single 400-mg oral dose of gatifloxacin and moxifloxacin . These samples were collected prior to and at 2, 6, 12, and 24 h after the dose of each drug . Gatifloxacin exhibited bactericidal activity for a median of 12 h against Streptococcus pneumoniae (MIC = 0.5 micro g/ml), Peptostreptococcus micros (MIC = 0.25 micro g/ml), and F . nucleatum (MIC = 0.5 micro g/ml) and 24 h against H . influenzae (MIC = 0.03 micro g/ml), Staphylococcus aureus (MIC = 0.125 micro g/ml), Peptostreptococcus magnus (MIC = 0.125 micro g/ml), and Prevotella melaninogenica (MIC = 0.5 micro g/ml) . Moxifloxacin exhibited bactericidal activity for a median of 24 h against Streptococcus pneumoniae (MIC = 0.125 micro g/ml), H . influenzae (MIC = 0.015 micro g/ml), Staphylococcus aureus (MIC = 0.06 micro g/ml), F . nucleatum (MIC = 0.5 micro g/ml), Prevotella melaninogenica (MIC =0.5 micro g/ml), Peptostreptococcus magnus (MIC = 0.125 micro g/ml), and Peptostreptococcus micros (MIC = 0.25 micro g/ml) . The results from this pharmacodynamic study suggest that these fluoroquinolones would have prolonged killing activity against these organisms in vivo and may have clinical utility in the treatment of mixed aerobic-anaerobic respiratory tract infections.

Dis Aquat Organ, 2003 Feb 13, 53(2), 107 - 13
Cloning and nucleotide sequence analysis of the chloramphenicol resistance gene on conjugative R plasmids from the fish pathogen Photobacterium damselae subsp . piscicida; Morii H et al.; Transferable resistance to various drugs was investigated in Photobacterium damselae subsp . piscicida from Japan . Drug resistances were transferred via plasmids of 100, 50, and 40 kb . Resistance to chloramphenicol (Cmr) was transferred on plasmids of all 3 sizes . The Cmr gene (cat) was cloned from the 50 kb plasmids pPDP8511 and pPDP9106 transferred from P . damselae subsp . piscicida strains isolated in different years and places in Japan . Subcloning localized the cat to within 1.5 kb HindIII-HincII (or PstI) fragments . Nucleotide sequences of the coding and flanking region of the cat were determined as 1607 bp (HindIII-HincII fragment) in pPDP8511 and 1568 bp (HindIII-PstI fragment) in pPDP9106, which corresponded with the sequence from nucleotides 40 to 1607 in pPDP8511 . The nucleotide sequences identified an open reading frame (ORF) encoding 213 amino acid residues with a calculated molecular mass of about 24.8 kDa, a size consistent with the molecular mass of known cat gene products, and the ORF had maximum homology (99.5%) with a Type II CAT variant from Haemophilus influenzae.

J Laryngol Otol, 2003 Mar, 117(3), 173 - 6
Failure of antibiotic therapy in acute otitis media; Babin E et al.; The aim of this retrospective study was to determine the possible causes of failure of antibiotic therapy in children with acute otitis media (AOM) . Thirty-nine samples of middle-ear fluid were obtained by myringotomy from 31 children suffering from AOM, unrelieved by antibiotic therapy administered for over 48 hours . The samples were analysed by the usual microbiological techniques, including cultures, tests for beta-lactamase producing strains and the determination of the minimal inhibitory concentration of penicillin for Streptococcus pneumoniae . In 14 samples, no bacterial strains were detected in the cultures of middle-ear fluid; and in two samples the cultures revealed two strains of bacteria . The bacteria most frequently identified were Haemophilus influenzae, found in 11 samples, and Streptococcus pneumoniae, found in seven samples, of which four produced strains with reduced susceptibility to penicillin . The failure of antibiotic therapy in AOM appears to be related to the increased resistance of Haemophilus influenzae and to the reduced susceptibility of Streptococcus pneumoniae to penicillin . Other factors contributing to the failure of antibiotic therapy in AOM may be the viruses or the bacteria that produce multiple pathogens in the middle ear.

J Laryngol Otol, 2003 Mar, 117(3), 169 - 72
Bacterial aetiology of non-resolving otitis media in South African children; Huebner RE et al.; Little is known of the aetiology, serotypes or susceptibility of the pathogens causing non-resolving otitis media in children receiving care from specialists in private practice in developed or in developing countries . Increased access to antibiotics in the community amongst children receiving such private care in South Africa may be anticipated to lead to levels of resistance similar to those found in countries with similar models of private practice, such as the United States . This study was conducted to determine the aetiology of non-resolving otitis media in South African children receiving private care and to determine the antimicrobial resistance patterns and serotypes of the bacterial isolates . Middle-ear fluid was cultured from 173 children aged two months to seven years with non-resolving acute otitis media accompanied by persistent pain or fever who were referred to otorhinolaryngologists for drainage of middle-ear fluid within 14 days of the start of symptoms . While 92 per cent of the children had recently received antibiotics and 54 per cent were currently receiving them, bacteria were isolated from 47 children (27 per cent) . Streptococcus pneumoniae was the most common pathogen (35), followed by Haemophilus influenzae (nine), Staphylococcus aureus (six), Moraxella catarrhalis (two), Streptococcus pyogenes (two) and Pseudomonas aeruginosa (one) . Two isolates were identified in each of eight children . Antimicrobial resistance to one or more antibiotics was found in 33/35 (94 per cent) of the pneumococci isolated, with resistance to penicillin in 86 per cent, resistance to trimethoprim-sulfamethoxazole in 54 per cent and to erythromycin and clindamycin in 69 per cent and 57 per cent, respectively . The pneumococcal serotypes found were 19F (28 per cent), 14 (26 per cent), 23F (23 per cent), 6B (nine per cent), 19A (87 per cent), and four (three per cent) . Children with a bacterial pathogen isolated were younger (mean age of 17 months) than children from whom no bacteria were isolated (mean age of 23 months; p = 0.03) . Isolation of a pneumococcus was also significantly associated with younger age (mean = 16 months versus 22 months, p = 0.03), the presence of fever (OR = 2.15, p = 0.049), and having one or more prior episodes of otitis media within the six months before tympanocentesis (OR = 7.72, p = 0.03) . Almost all pneumococci isolated from non-resolving acute otitis media in this community are antibiotic-resistant and should be considered especially in young children who have failed previous therapy and who have non-resolving pain or fever.

Biophys Chem, 2003, 100(1-3), 437 - 52
Isolation and characterization of the prokaryotic proteasome homolog HslVU (ClpQY) from Thermotoga maritima and the crystal structure of HslV; Song HK et al.; Heat-shock locus VU (HslVU) is an ATP-dependent proteolytic system and a prokaryotic homolog of the proteasome . It consists of HslV, the protease, and HslU, the ATPase and chaperone . We have cloned, sequenced and expressed both protein components from the hyperthermophile Thermotoga maritima . T . maritima HslU hydrolyzes a variety of nucleotides in a temperature-dependent manner, with the optimum lying between 75 and 80 degrees C . It is also nucleotide-unspecific for activation of HslV against amidolytic and caseinolytic activity . The Escherichia coli and T . maritima HslU proteins mutually stimulate HslV proteins from both sources, suggesting a conserved activation mechanism . The crystal structure of T . maritima HslV was determined and refined to 2.1-A resolution . The structure of the dodecameric enzyme is well conserved compared to those from E . coli and Haemophilus influenzae . A comparison of known HslV structures confirms the presence of a cation-binding site, although its exact role in the proteolytic mechanism of HslV remains unclear . Amongst factors responsible for the thermostability of T . maritima HslV, extensive ionic interactions/salt-bridge networks, which occur specifically in the T . maritima enzyme in comparison to its mesophilic counterparts, seem to play an important role.

Nihon Kokyuki Gakkai Zasshi, 2002 Nov, 40(11), 905 - 9
{A case of AIDS-associated Haemophilus influenzae pneumonia with diffuse reticulonodular shadows}; Konishi M et al.; A 32-year-old male was admitted to our hospital complaining of fever and dyspnea on effort . Laboratory data on admission indicated leukocytosis and elevation of C-reactive protein . A chest radiograph showed diffuse reticulonodular shadows in both lower lung fields, and a chest computed tomography showed centrilobular reticulonodular opacity . Bronchoscopic findings revealed a large amount of slightly yellowish secretion in all bronchi . Cells found in the bronchoalveolar lavage fluid (BALF) included 61% neutrophils . Haemophilus influenzae was isolated from cultures of the BALF and sputum . Transtracheal lung biopsy specimens showed focal infiltration of neutrophils in the alveoli, and the pathological findings in the lung were compatible with bronchiolopneumonia . Since the CD4/CD8 ratio was 0.09 and a positive reaction was obtained for anti-human immunodeficiency virus (HIV) antibody, HIV-associated pneumonia due to H . influenzae was diagnosed . Seven days' administration of cefozopran improved the patient's condition . It is interesting that radiological findings are often unusual in HIV-infected patients with H . influenzae pneumonia.

Plant Physiol, 2003 Mar, 131(3), 1209 - 19
Arabidopsis proteins containing similarity to the universal stress protein domain of bacteria; Kerk D et al.; We have collected a set of 44 Arabidopsis proteins with similarity to the USPA (universal stress protein A of Escherichia coli) domain of bacteria . The USPA domain is found either in small proteins, or it makes up the N-terminal portion of a larger protein, usually a protein kinase . Phylogenetic tree analysis based upon a multiple sequence alignment of the USPA domains shows that these domains of protein kinases 1.3.1 and 1.3.2 form distinct groups, as do the protein kinases 1.4.1 . This indicates that their USPA domain structures have diverged appreciably and suggests that they may subserve distinct cellular functions . Two USPA fold classes have been proposed: one based on Methanococcus jannaschii MJ0577 (1MJH) that binds ATP, and the other based on the Haemophilus influenzae universal stress protein (1JMV), highly similar to E . coli UspA, which does not bind ATP . A set of common residues involved in ATP binding in 1MJH and conserved in similar bacterial sequences is also found in a distinct cluster of Arabidopsis sequences . Threading analysis, which examines aspects of secondary and tertiary structure, confirms this Arabidopsis sequence cluster as highly similar to 1MJH . This structural approach can distinguish between the characteristic fold differences of 1MJH-like and 1JMV-like bacterial proteins and was used to assign the complete set of candidate Arabidopsis proteins to one of these fold classes . It is clear that all the plant sequences have arisen from a 1MJH-like ancestor.

J Infect, 2003 Apr, 46(3), 194 - 6
Rapidly fatal Haemophilus influenzae serotype f sepsis in a healthy child; Zacharisen MC et al.; A previously healthy 4-year-old child became acutely ill with vomiting and low-grade fever . The following day she suddenly became limp and unresponsive . She experienced acute septic shock and despite aggressive treatment died . Blood cultures grew ampicillin-resistant Haemophilus influenzae type f . There was no evidence of bacterial pneumonia or meningitis . To our knowledge, this represents the first case of fatal H . influenzae type f sepsis in a child without an identifiable focus or underlying predisposing condition . Despite the overwhelming success of the H . influenzae type b vaccine, physicians need to be aware of the potential for severe and fatal H . influenzae infections other than type b.

J Infect, 2003 Apr, 46(3), 191 - 4
Haemophilus paraphrophilus prosthetic valve endocarditis; Watkin RW et al.; We report a case of prosthetic, aortic valve, infective endocarditis caused by Haemophilus paraphrophilus . There are no other cases described in the available literature where this microorganism has caused prosthetic valve endocarditis and no other case reported involving only the aortic valve.

Rev Med Chir Soc Med Nat Iasi, 2002 Apr-Jun, 107(2), 348 - 51
{Hemophilus influenzae meningitis- the experience of infectious diseases department between 1984-2001}; Corcaci C et al.; Our paper presents a clinico-biological and therapeutical study of the cases admitted in Infectious Diseases Hospital between January 1984-December 2001 . We studied the records of all the patients that suffered from meningitis with Haemophilus influenzae . In above mentioned period 40 cases of Haemophilus influenzae meningitis was admitted; 6 patients (15%) died . A number of 13 cases was registered for the first five years, mainly in the childhood-32 cases (80%) for infants . Bacteriological diagnosis was made by bacterioscopia for 16 patients, bacteriological cultures-30 patients, latex test, performed in the last five years, in 8 cases . In patients who died, the bacteriologic culture was positive in 5 cases and bacterioscopia in 4 cases . The therapy was performed in resonance with antibiograme results for the involved strains . The Haemophilus influenzae meningitis is an acute and severe disease of the childhood, with an important risk of death, despite the rigorous therapeutical measures.

Kansenshogaku Zasshi, 2003 Jan, 77(1), 1 - 4
{The frequency of non-typeable Haemophilus influenzae systemic disease in children}; Ishiwada N et al.; 95 strains of Haemophilus influenzae (H . influenzae) isolated from blood of the patients with systemic infections were serotyped by staphylococcal coagglutination during the ten years from 1992 through 2001 . As a result, 92 (96.8%) cases were caused by type b strains and 3 (3.2%) cases were caused by non-typeable strains . Three cases with systemic infection due to non-typeable H . influenzae were reported . One patient was a premature neonate with sepsis and respiratory failure who had a fulminant course and died . The other two patients were a 3-year-old girl and a 1-month-old boy both with pneumonia . About their underlying conditions, one received intravenous steroid therapy and the other suffered from respiratory syncytial virus infection . They were treated with appropriate antibiotics and their clinical courses were satisfactory and uncomplicated . Non-typeable H . influenzae was isolated from not only blood but also the lower respiratory tract in all three cases . Systemic infection due to non-typeable strain is rare . But, it should be recognized as a substantial proportion of the serious infections caused by H . influenzae.

Clin Ther, 2003 Jan, 25(1), 169 - 77
Activity of nine oral agents against gram-positive and gram-negative bacteria encountered in community-acquired infections: use of pharmacokinetic/pharmacodynamic breakpoints in the comparative assessment of beta-lactam and macrolide antimicrobial agents; Peric M et al.; BACKGROUND: The application of pharmacokinetic (PK) and pharmacodynamic (PD) data in conjunction with minimum inhibitory concentrations (MICs) of antibacterial agents has been shown to allow for improved selection and appropriate dosing of antimicrobial agents for specific infections, increasing the likelihood of bacteriologic cure and, through this, reducing the risk for the development of resistant organisms . OBJECTIVES: This study was undertaken to provide data on current levels of resistance among common community-acquired bacterial species to 7 betalactam antimicrobial agents (including the combination product amoxicillin/clavulanate), azithromycin, and clarithromycin, determined through application of the PK/PD breakpoints based on time-above-MIC for the beta-lactams and the nonazalide macrolide clarithromycin, and on 24-hour serum area under the curve divided by MIC for the azalide macrolide azithromycin . METHODS: The antimicrobial products tested were amoxicillin/clavylanate, cefpodoxime, cefdinir, cefditoren, cefprozil, cefuroxime, cefixime, azithromycin, and clarithromycin . The bacterial species comprised 70 penicillin-susceptible, 68 penicillin-intermediate, and 69 penicillin-resistant strains of Streptococcus pneumoniae; 46 beta-lactamase-positive and 54 beta-lactamase-negative strains of Haemophilus influenzae; 49 strains of Moraxella catarrhalis; and 100 methicillin-sensitive strains of Staphylococcus aureus (MSSA) . Strains were isolated from clinical specimens obtained from outpatient-acquired infections in 1 clinical center in the Northeast and 1 in the north-central area of the United States within the past 2 years . National Committee for Clinical Laboratory Standards microdilution MIC methodology was used . PK/PD breakpoints were obtained from previously published studies and were based on blood values . RESULTS: Amoxicillin/clavulanate was the product to which the greatest percentage of susceptible, intermediate, and resistant strains of pneumococci were sensitive at the PK/PD breakpoint, followed by cefditoren, cefpodoxime, cefuroxime, cefdinir, and cefprozil . None of the cephalosporins were active against penicillin-resistant pneumococci . Cefditoren and cefpodozime were the agents to which the greatest percentage of beta-lactamase-positive and beta-lactamase-negative strains of H influenzae were sensitive, followed by amoxicillin/clavulanate, cefdinir, and cefuroxime . Cefprozil was inactive against H influenzae . All of the beta-lactam products were active against M catarrhalis . All but cefpodoxime, cefditoren, and cefixime were active against MSSA . CONCLUSIONS: In this study, based on PK/PD breakpoints, amoxicillin/clavulanate had the best overall activity of the 9 antimicrobial products tested . Cefpodoxime and cefditoren were active against >or=90% of strains of penicillin-susceptible and penicillin-intermediate pneumococci, H influenzae, and M catarrhalis . The macrolides azithromycin and clarithromycin were active against penicillin-susceptible and penicillin-intermediate pneumococci and M catarrhalis; they were inactive against H influenzae and penicillin-resistant pneumococci.

Pediatr Infect Dis J, 2003 Mar, 22(3), 262 - 8
Differences in nasopharyngeal bacterial flora in children with nonsevere recurrent acute otitis media and chronic otitis media with effusion: implications for management; Marchisio P et al.; BACKGROUND: The interactions between nasopharyngeal flora and the individual entities covered by the broad term otitis media have not been completely elucidated . We investigated in infants and children ages 6 months to 7 years with nonsevere recurrent acute otitis media (rAOM) or with chronic otitis media with effusion (cOME): (1) the nasopharyngeal carriage rate and bacterial density of respiratory pathogens and alpha-hemolytic streptococci in comparison with healthy children; (2) the resistance pattern of respiratory pathogens; and (3) the relationship between the type of nasopharyngeal colonization and long term outcome . METHODS: Nasopharyngeal cultures were obtained from 85 children with rAOM,113 children with cOME and 55 controls . A semiquantitative analysis was used in the reading of cultures . A 12-week follow-up without treatment was planned . RESULTS: The carrier rate of respiratory pathogens was significantly greater in cOME (70%) than in rAOM (45%) (P = 0.0006) or controls (31%) (P < 0.0001) . Similarly colonization density was significantly greater in cOME than in rAOM . The carriage rate and the colonization density of alpha-hemolytic streptococci were significantly lower in rAOM than in cOME or controls . The incidence of resistant (R) strains was greater in rAOM (Streptococcus pneumoniae penicillin-R, 24%; macrolide-R, 64%; Haemophilus influenzae amoxicillin-R, 24%) compared with cOME (S . pneumoniae penicillin-R,18%; macrolide-R, 44%; H . influenzae amoxicillin-R, 5%) or controls (S . pneumoniae penicillin-R, 8%; macrolide-R, 23%; H . influenzae amoxicillin-R, 10%) . During the follow-up period persistence of OME and occurrence of AOM were greater among carriers of respiratory pathogens at baseline . CONCLUSIONS: There are substantial differences in nasopharyngeal flora between children with nonsevere rAOM and children with cOME . The results of nasopharyngeal cultures should be taken into account to avoid treatment with drugs that are ineffective and likely to select resistant organisms.

Pediatr Infect Dis J, 2003 Mar, 22(3), 209 - 16
Recurrent acute otitis media occurring within one month from completion of antibiotic therapy: relationship to the original pathogen; Leibovitz E et al.; OBJECTIVES: (1) To determine the relationship between acute otitis media (AOM) pathogens isolated in cases of early clinical recurrence of AOM (occurring within 1 month from completion of therapy) to the original pathogens causing the initial AOM episode; and (2) To determine whether shorter time intervals between completion of antibiotic therapy and clinical recurrences of AOM are associated with higher rates of true bacteriologic relapse . PATIENTS AND METHODS: From 1995 through 2000, 1077 infants and young children ages 3 to 36 months with AOM were enrolled in double tympanocentesis (performed on Day 1 in all patients and Days 4 to 6 in those initially culture-positive) studies . Of these, 834 (77%) completed successfully the antibiotic treatment {pathogen eradication on Days 4 to 6 of therapy or no pathogen on middle ear fluid (MEF) culture on Day 1 and clinical improvement at end of therapy} . Patients were followed for 3 to 4 weeks after completion of therapy, and additional MEF cultures were obtained if clinical recurrence occurred . True bacteriologic relapse was defined as the presence of a pathogen identical with that isolated before therapy by serotype and pulsed field gel electrophoresis for and by pulsed field gel electrophoresis for Streptococcus pneumoniae and beta-lactamase production for Haemophilus influenzae . RESULTS: MEF cultures were performed in 108 consecutive patients with early recurrent AOM . One hundred pathogens were isolated at recurrence in 88 of 108 (81%) patients: 54 H . influenzae; 45 S . pneumoniae; and 1 Moraxella catarrhalis . Most recurrent AOM episodes developed during the first 2 weeks of follow-up; 39 (36%), 38 (35%), 21 (19%) and 10 (9%) recurrent AOM episodes occurred on Days 1 to 7, 8 to 14, 15 to 21 and 22 to 28 after completion of therapy, respectively . In most patients these episodes were caused by a new pathogen . True bacteriologic relapses were found in 30 (28%) of 108 patients whose MEF cultures were positive for 35 pathogens: 13 of 108 (12%) S . pneumoniae; 12 of 108 (11%) H . influenzae; and 5 of 108 (5%) both . When timing of recurrent AOM after completion of therapy was analyzed, true bacteriologic relapses were found in 16 of 39 (41%), 10 of 38 (26%), 3 of 21 (14%) and 1 of 10 (10%) of all episodes on Days 1 to 7, 8 to 14, 15 to 21 and 22 to 28 after completion of therapy, respectively (P = 0.01) . The respective rates for were 11 of 17 (65%), 3 of 10 (30%), 3 of 13 (23%) and 1 of 5 (20%) (P = 0.02) . For H . influenzae the respective rates were 8 of 19 (42%), 9 of 23 (39%), 0 of 8 (0%) and 0 of 4 (0%) (P = 0.02) . CONCLUSIONS: Most recurrent AOM episodes occurring within 1 month from completion of antibiotic therapy are in fact new infections . Most of the true bacteriologic AOM relapses occur within 14 days after completion of therapy, but even during this time interval most of the recurrences are caused by new pathogens . H . influenzae is very unlikely to cause true bacteriologic AOM relapses 14 days or later after completion of therapy.

Arch Neurol, 2003 Mar, 60(3), 431 - 3
Late-developing cerebral arteropathy after pyogenic meningitis; Palacio S et al.; BACKGROUND: Although vasculopathy is a recognized complication during acute meningitis, to our knowledge, no previous reports have been published of this phenomenon developing months after successful treatment . OBJECTIVE: To report a unique case of a late-developing vasculopathy after pyogenic meningitis in an adult . REPORT OF A CASE: A 51-year-old woman was seen with severe headache 2 months after treatment of Haemophilus influenzae type C meningitis . Initial arteriography showed no abnormality; a second arteriogram showed progressive multifocal intracranial stenosis affecting mainly the internal carotid arteries . Findings from pathologic examination disclosed diffuse collagenosis consistent with chronic vascular injury from meningitis . The arterial lesions stabilized, and the patient remained asymptomatic . CONCLUSION: Progressive intracranial arterial stenosis can evolve months after meningitis and should be added to the list of recognized vascular complications.

Microb Pathog, 2003 Mar, 34(3), 131 - 9
Genetic manipulation of immunoglobulin binding proteins of Haemophilus somnus; Sanders JD et al.; The relationship of the 76kDa immunoglobulin binding, surface antigen (p76) of Haemophilus somnus to the high molecular weight immunoglobulin binding proteins (HMW IgBPs) was investigated . The kanamycin resistance gene from pLS88 was used via homologous recombination with allelic exchange to replace a portion of the gene encoding IgBPs of H . somnus strain 8025 . Recombinants were shown by Western immunoblotting to express and secrete truncated antigens of approximately 200kDa and not to produce p76 . The truncated HMW IgBP variants retained the ability to bind bovine IgG2 by the Fc portion as demonstrated by Western immunoblotting against IgG2 anti-DNP . This data indicated that the deleted 1.8kb BglII fragment was not required for secretion or immunoglobulin Fc binding by the HMW IgBPs but was required for expression of the downstream p76 gene . Functional studies showed that, in addition to Fc binding of IgG2 to truncated HMW IgBPs, the mutant strain 8025 Kan1 was equally resistant to killing by mouse complement but less virulent than the wild type parent (8025) in a mouse septicemia model of H . somnus infection . However, mutant strain 8025 Kan1 did adhere less well than the wild type to bovine pulmonary artery endothelial cells . It is probable that p76 and the missing peptides of the HMW IgBPs play a role in this aspect of virulence and perhaps other aspects.

Microb Pathog, 2003 Mar, 34(3), 121 - 30
Involvement of lipooligosaccharides of Haemophilus influenzae and Neisseria meningitidis in defensin-enhanced bacterial adherence to epithelial cells; Gorter AD et al.; Stimulated neutrophils release a variety of antimicrobial peptides, including neutrophil defensins (HNP1-4) . We have previously reported that neutrophil defensins enhanced the adherence of Haemophilus influenzae and Neisseria meningitidis to cultured respiratory epithelial cells . In this study, the effect of defensins on the adherence of H . influenzae and N . meningitidis lipooligosaccharide (LOS) mutants to epithelial cells was tested . Neutrophil defensins enhanced the adherence of the oligosaccharide mutants of H . influenzae and N . meningitidis, whilst the adherence of the lipid A mutants B29 of H . influenzae and lpxL1 and lpxL2 of N . meningitidis was not or only moderately stimulated by neutrophil defensins . The adherence of the N . meningitidis LOS negative mutant lpxA was not enhanced by defensins . These findings suggested that the secondary fatty acids of lipid A were involved in the defensin-enhanced adherence . LOS from strain H44/76 or HNP-LOS complexes did not affect or stimulate the adherence of N . meningitidis, although the defensin-enhanced adherence is specific for certain bacterial species having LOS in their outer membrane . These results indicated that LOS is involved in the defensin-enhanced adherence . However, the mechanism by which defensins and LOS interact with epithelial cells to promote bacterial adherence remains to be resolved.

J Trop Pediatr, 2003 Feb, 49(1), 28 - 32
Percentage, bacterial etiology and antibiotic susceptibility of acute respiratory infection and pneumonia among children in rural Senegal; Echave P et al.; Acute respiratory infections (ARI) are still a major health problem in most developing countries . So far no study has evaluated the importance of childhood ARI in rural Senegal . We prospectively studied ARI, the percentage of pneumonia and related mortality, as well as the bacterial composition of nasopharyngeal flora using nasopharyngeal aspirates in 114 children, aged 2-59 months, presenting at Ndioum's pediatric ward . Excluded from the trial were those children that had had antimicrobial therapy in the previous 2 weeks . The Kirby-Bauer method was used to determine antibiotic resistance throughout the study . The percentage of ARI and pneumonia among the population tested was 24 per cent and 11 per cent respectively . Streptococcus pneumonia was often resistant to cotrimoxazole (31 per cent) but only 9 per cent were resistant to chloramphenicol and 14 per cent to penicillin . Haemophilus influenzae (HI) was uniformly sensitive to ampicillin, and only 4 per cent were resistant to chloramphenicol and 11 per cent to cotrimoxazole . We conclude that SP and HI resistance to cotrimoxazole is important and warrants larger clinical trials using chloramphenicol . Information campaigns and intense management of comorbidities are desirable in this type of population . Comorbidities (tuberculosis, malaria, HIV-AIDS, severe malnutrition) are determinant variables in many ARI cases and carry a high negative prognosis value.

East Afr Med J, 2002 Nov, 79(11), 588 - 92
Antibacterial effect of Zingiber officinale and Garcinia kola on respiratory tract pathogens; Akoachere JF et al.; OBJECTIVE: To investigate the antibacterial activity of Zingiber officinale (ginger) Garcinia kola (bitter kola) on four respiratory tract pathogens . DESIGN: A prospective study based on laboratory investigations . SETTING: Department of Life Sciences, University of Buea . Throat swabs were collected from 333 individuals with running nostrils, cough and/or catarrh in three localities of Buea namely Bokwango, Molyko and Bolifamba . Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae and Haemophilus influenzae were isolated from the specimens using standard microbiological procedures . The antibacterial activity of ethanolic extracts of ginger and bitter kola, were investigated on these pathogens using the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays . RESULTS: The extracts exhibited antibacterial activity against the pathogens . The MIC of extracts ranged from 0.0003 microg/ml to 0.7 microg/ml for ginger and 0.00008 microg/ml, to 1.8 microg/mL for bitter kola, while MBC ranged from 0.1.35 microg/ml to 2.04 microg/ml for ginger and 0.135 microg/ml to 4.2 microg/ml for bitter kola . CONCLUSION: Results indicated that extracts of ginger root and bitter kola may contain compounds with therapeutic activity.

Clin Diagn Lab Immunol, 2003 Mar, 10(2), 202 - 7
Levels of antibodies specific to tetanus toxoid, Haemophilus influenzae type b, and pneumococcal capsular polysaccharide in healthy children and adults; Schauer U et al.; Antibody levels specific for capsular polysaccharides of Streptococcus pneumoniae and Haemophilus influenzae type b (Hib) and for tetanus toxoid were measured in serum samples of 386 age-stratified subjects . The study group consists of healthy adult blood donors and hospitalized children undergoing elective surgery, excluding individuals with a history of infection . In children, anti-tetanus toxoid antibody levels displayed two peaks of 1.20 IU/ml (20.4 mg/liter) and 1.65 IU/ml (28.1 mg/liter) related to the schedule of routine childhood immunization in the first year and at 8 years of age . Eighty percent of the antibodies are of the immunoglobulin G1 (IgG1) isotype . For pneumococcal capsular polysaccharide (PCP), the specific antibody levels represent the acquisition of natural immunity . The initial concentration of 9.2 mg/liter was low in infancy (0.5 to 1 years of age) and remained low until 3 to 4 years of age (14.6 mg/liter) . During this period PCP antibodies were almost 100% of the IgG2 subclass . Thereafter, IgG anti-PCP antibody titers increased steadily to adult levels (59.5 mg/liter) . The data are intended to provide reference ranges to aid in the interpretation of specific antibody determinations in the clinical setting.

OMICS, 2002, 6(4), 341 - 8
H . influenzae Consortium: integrative study of H . influenzae-human interactions; Kolker E et al.; Developments in high-throughput analysis tools coupled with integrative computational techniques have enabled biological studies to reach new levels . The ability to correlate large volumes of diverse data types into cohesive models of organism function has spawned a new systematic approach to biological investigation . The creation of a new consortium has been proposed to investigate a single organism utilizing these comprehensive approaches . The Haemophilus influenzae Consortium (HIC) would be comprised of five laboratories, each providing separate and complementary areas of expertise in the study of Haemophilus influenzae (HI) . The 5-year study proposes to develop coherent models of HI, both as a stand-alone organism, and more importantly, as a human pathogen . Studies in growth condition specificity followed by genomic, metabolic, and proteomic experimentation will be combined and integrated through computational and experimental analyses to form dynamic and predictive models of HI and its responses . Data from the HIC will allow greater understanding of cellular behavior, pathogen-host interactions, bacterial infection, and provide future scientific endeavors with a template for studies of other pathogens.

Infect Dis Obstet Gynecol, 2002, 10(3), 161 - 4
Haemophilus influenzae septic abortion; Cherpes TL et al.; BACKGROUND: Haemophilus influenzae septic abortion is typically caused by nontypeable strains of the organism . Furthermore, nontypeable species with a special affinity for the genital tract are the most frequent isolates encountered, and an ascending vaginal or cervical infection is often the suspected route of transmission . CASE: A 39-year-old woman at 8 weeks gestation who underwent dilation, evacuation, and curettage for embryonic demise had clinical evidence for sepsis and isolation of a nontypeable, ampicillin resistant H . influenzae from blood cultures . Although an ascending vaginal infection was suspected, the route of transmission was not determined . CONCLUSION: Nontypeable strains of . H . influenzae have demonstrated increased beta-lactamase activity, and ampicillin, formerly the treatment of choice, should be used only if isolate susceptibility is known.

Biologicals, 2003 Mar, 31(1), 39 - 43
Identification and characterization by 16S rDNA analysis of viable bacterial colonies isolated from oral medicines based on inactivated or lysed pathogenic bacteria; Studer E et al.; Oral bacterial immunomodulators are based on inactivated or lysed pathogenic bacterial cells . The safety of these products for consumers critically depends on the effectiveness of procedures used for pathogen inactivation . In a market survey in Switzerland we tested 26 lots of three different immunomodulators for the presence of any remaining culturable cells . Dissolved stimulants were plated on Eugon agar for the unspecific cultivation of bacteria (including most of the pathogenic bacteria in the modulator) and on Chocolate+PolyViteX agar for the cultivation of Haemophilus influenzae . A total of 16 colonies were grown on either Eugon agar or Chocolate+PolyViteX agar . These colonies were characterized by amplifying and sequencing a 16S rDNA fragment using unspecific screening primers . None of the sequenced fragments could be associated with the inactivated or lysed pathogenic bacteria present in the modulator . These data indicate that the pathogen inactivation procedures used for all tested products are effective . They also demonstrate full compliance of all products with pharmacopoeial requirements regarding microbial purity . Finally, the spectrum of germs isolated confirms the notion that man is the primary source of microbial contamination in pharmaceutical products.

Jpn J Antibiot, 2002 Dec, 55(6), 861 - 5
{Meropenem was effective to the bacterial meningitis due to beta-lactamase negative ampicillin-resistant Haemophilus influenzae; case report}; Otsuka T et al.; We experienced a case in the infant of the bacterial meningitis due to beta-lactamase negative ampicillin-resistant Haemophilus influenzae (BLNAR) which has increased in recent years . In the present condition, the sensitivity of the bacteria to the antibacterial-drug used as the initial treatment for the bacterial meningitis is sometimes inadequate . If it takes into consideration that BLNAR participates in H . influenzae meningitis, it will be one of the choice to use meropenem with other antibacterial-drug concomitantly.

Jpn J Antibiot, 2002 Dec, 55(6), 808 - 26
{Antibacterial activity of panipenem against clinical isolates in 2000 and 2001}; Abe T et al.; As the post-marketing surveillance of panipenem/betamipron (Carbenin), MICs of panipenem (PAPM) against 1355 clinical isolates of 28 species from 15 medical institutions all over Japan from June 2000 to March 2001 were measured using the broth microdilution method approved by the Japanese Society of Chemotherapy and compared with those of parenteral carbapenem antibacterials, imipenem (IPM) and meropenem (MEPM), and parenteral cephem antibacterials, cefozopran, cefepime, and sulbactam/cefoperazone . The activity of PAPM was comparable to that of IPM against almost all species tested . Compared with MEPM, PAPM was more active against Gram-positive bacteria and Bacteroides spp., and less active against Gram-negative bacteria . Compared with the parenteral cephems, PAPM was more active against most of species tested and its MIC ranges were narrower than those of the cephems as were those of other carbapenems . In this surveillance study, the incidence of resistance in various species were as follows: 39.3% for methicillin-resistant Staphylococcus aureus, 47.3% for penicillin-intermediate Streptococcus pneumoniae (PISP), 15.1% for penicillin-resistant S . pneumoniae (PRSP), 0.9% for extended-spectrum beta-lactamase (ESBL) producing Escherichia coli, 3.4% for ESBL producing Klebsiella pneumoniae, 19.2% for beta-lactamase producing Haemophilus influenzae, 24.0% for beta-lactamase-negative ampicillin-resistant (BLNAR) H . influenzae, and 1.0% for metallo-beta-lactamase producing Pseudomonas aeruginosa . Against these resistant strains, carbapenems including PAPM showed generally more potent activity than cephems . It was noted that PAPM showed the most potent activity against PISP and PRSP, which showed high incidence of 62.4% totally, among tested drugs . Metallo-beta-lactamase producing P . aeruginosa exhibited high resistance and BLNAR H . influenzae also exhibited low susceptibility against all tested drugs . But no remarkable change in the activity of PAPM against other species was observed in this study compared with that in the studies before the marketing of Carbenin . Furthermore, it is necessary to pay much attention to the trend of resistant strains such as PRSP, metallo-beta-lactamase producing bacteria, and BLNAR H . influenzae.

Arch Ophthalmol, 2003 Mar, 121(3), 345 - 50
Vitreous and aqueous penetration of orally administered gatifloxacin in humans; Hariprasad SM et al.; OBJECTIVE: To investigate the penetration of gatifloxacin, a novel extended-spectrum fourth-generation fluoroquinolone antibiotic, into the vitreous and aqueous humor after oral administration . METHODS: A prospective, nonrandomized study of 24 consecutive patients scheduled for pars plana vitrectomy between September 2001 and May 2002 at the Cullen Eye Institute . Aqueous, vitreous, and serum samples were obtained and analyzed from 24 patients after administration of two 400-mg gatifloxacin tablets taken 12 hours apart before the operation . Assays were performed using high-performance liquid chromatography . RESULTS: Mean +/- SD gatifloxacin concentrations in serum (n = 23), vitreous (n = 23), and aqueous (n = 11) were 5.14 +/- 1.36 micro g/mL, 1.34 +/- 0.34 micro g/mL, and 1.08 +/- 0.54 micro g/mL respectively . Mean +/- SD sampling times after oral administration of the second gatifloxacin tablet for serum, vitreous, and aqueous were 3.2 +/- 1.0 hours, 4.0 +/- 1.0 hours, and 3.9 +/- 1.1 hours, respectively . The percentages of serum gatifloxacin concentration achieved in the vitreous and aqueous were 26.17% and 21.02%, respectively . Mean inhibitory vitreous and aqueous MIC(90 ) levels were achieved against many pathogens, including Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Propionibacterium acnes, Haemophilus influenzae, Escherichia coli, Bacillus cereus, Proteus mirabilis, and other organisms . CONCLUSIONS: Orally administered gatifloxacin achieves therapeutic levels in the noninflamed human eye, and the activity spectrum appropriately encompass the bacterial species most frequently involved in the various causes of endophthalmitis . Because of its broad-spectrum coverage, low MIC(90) levels for the organisms of concern, and good tolerability, gatifloxacin represents a major advance in the prophylaxis or treatment of postoperative, posttraumatic, and bleb-associated bacterial endophthalmitis.

Neurol Clin, 2002 Nov, 20(4), 1013 - 38, vi-vii
Meningitis and encephalitis in children . An update; Bonthius DJ et al.; Over the course of the past decade, much has changed on the landscape of meningitis and encephalitis in children . West Nile virus has emerged in the United States as a new etiologic pathogen causing encephalitis . Human herpesvirus-6 has been identified as a cause of encephalitis and febrile seizures . Lymphocytic choriomeningitis virus has been identified as an underrecognized neuroteratogen . The emergence of penicillin-resistant Streptococcus pneumoniae has complicated the treatment of bacterial meningitis, whereas the Haemophilus influenzae vaccine has fundamentally altered the disease's epidemiology . The recognition that much of the neuropathologic change induced by bacterial meningitis is inflammation mediated has paved the way to the demonstration that dexamethasone can substantially improve the outcome of bacterial meningitis in children . Although much progress has been made toward understanding, treating, and preventing these important infections, much remains to be learned.

Sex Transm Dis, 2003 Mar, 30(3), 241 - 5
Association between HIV-1 infection, the etiology of genital ulcer disease, and response to syndromic management; Moodley P et al.; BACKGROUND: Reports on the effect of HIV-1 infection on healing rates of ulcers are conflicting . GOAL: The goal was to determine the etiology and response to treatment of genital ulcer disease (GUD) in relation to HIV-1 infection . STUDY DESIGN: This was a cohort study of patients with GUD treated with local syndromic management protocols . RESULTS: Among the 587 recruited, the prevalences of infections due to HSV, Treponema pallidum, Chlamydia trachomatis (lymphogranuloma venereum {LGV}), Haemophilus ducreyi, Calymmatobacterium granulomatis, and HIV-1 were 48%, 14%, 11%, 10%, 1%, and 75%, respectively . The prevalence T . pallidum of was higher among men (P = 0.03), and an association was seen among HIV-1-seronegatives on univariate and multivariate analyses (P < 0.001; = 0.01) . The prevalence of C trachomatis (LGV) was higher among females (P = 0.004), and an association was seen among HIV-1-seropositives on univariate analysis (P = 0.04) . At follow-up, 40/407 (10%) showed a decreased healing tendency, not associated with ulcer etiology or HIV-1 seropositivity . CONCLUSION: Response to syndromic management of GUD was acceptable and not associated with HIV-1 coinfection.

Proc Natl Acad Sci U S A, 2003 Mar 18, 100(6), 3089 - 94 Epub 2003 Mar 03.
Metabolic incorporation of unnatural sialic acids into Haemophilus ducreyi lipooligosaccharides; Goon S et al.; The lipooligosaccharides (LOS) of Haemophilus ducreyi are highly sialylated, a modification that has been implicated in resistance to host defense and in virulence . In previous work, we demonstrated that H . ducreyi scavenges sialic acid from the extracellular milieu and incorporates those residues into LOS . Here we report that H . ducreyi can use unnatural sialic acids bearing elongated N-acyl groups from three to seven carbon atoms in length, resulting in outer membrane presentation of unnatural sialyl-LOS . The unnatural variant comprises approximately 90% of cell surface sialosides when exogenous substrates are added to the media at micromolar concentrations, despite the availability of natural sialic acid in the growth media . Although they represent the majority of cell surface sialosides, analogs with longer N-acyl groups diminish the overall level of LOS sialylation, culminating in complete inhibition of LOS sialylation by N-octanoyl sialic acid . Thus, sialylation of H . ducreyi LOS can be modulated with respect to the structure of the terminal sialic acid residue and the extent to which the LOS acceptor is modified by supplying the bacteria with various sialic acid analogs.

J Antimicrob Chemother, 2003 Mar, 51(3), 727 - 30
Concentrations of garenoxacin in plasma, bronchial mucosa, alveolar macrophages and epithelial lining fluid following a single oral 600 mg dose in healthy adult subjects; Andrews J et al.; A microbiological assay was used to measure concentrations of garenoxacin (BMS-284756) in plasma, bronchial mucosa (BM), alveolar macrophages (AM) and epithelial lining fluid (ELF), following a single 600 mg oral dose . Twenty-four healthy subjects were allocated into four nominal time intervals after the dose, 2.5-3.5, 4.5-5.5, 10.5-11.5 and 23.5-24.5 h . Mean concentrations in plasma, BM, AM and ELF, respectively, for the four nominal time windows were for 2.5-3.5 h 10.0 mg/L (S.D . 2.8), 7.0 mg/kg (S.D . 1.3), 106.1 mg/L (S.D . 60.3) and 9.2 mg/L (S.D . 3.6); 4.5-5.5 h 8.7 mg/L (S.D . 2.2), 6.0 mg/kg (S.D . 1.9), 158.6 mg/L (S.D . 137.4) and 14.3 mg/L (S.D . 8.2); 10.5-11.5 h 6.1 mg/L (S.D . 1.9), 4.0 mg/kg (S.D . 1.4), 76.0 mg/L (S.D . 47.7) and 7.9 mg/L (S.D . 4.6); and 23.5-24.5 h 2.1 mg/L (S.D . 0.5), 1.7 mg/kg (S.D . 0.7), 30.7 mg/L (S.D . 12.9) and 3.3 mg/L (S.D . 2.3) . Concentrations at all sites exceeded MIC(90)s for the common respiratory pathogens Haemophilus influenzae (0.03 mg/L), Moraxella catarrhalis (0.015 mg/L) and Streptococcus pneumoniae (0.06 mg/L) . These data suggest that garenoxacin should be effective in the treatment of community-acquired pneumonia and chronic obstructive pulmonary disease.

J Antimicrob Chemother, 2003 Mar, 51(3), 523 - 30
Cefuroxime resistance in non-beta-lactamase Haemophilus influenzae is linked to mutations in ftsI; Straker K et al.; The penicillin binding protein (PBP) genes dacA, dacB and ftsI from 14 cefuroxime-resistant (CXM(R)) isolates and three clinical isolates with low CXM MIC for non-beta-lactamase-producing Haemophilus influenzae type b were molecularly characterized . One strain, 5788, was used to transform H . influenzae Rd to CXM(R) for direct comparison of the pbps in the same genetic background . No obvious mutations in the dacA and dacB gene products could be associated with CXM(R) . One amino acid substitution in the ftsI gene product in particular, S357N, could give rise to CXM(R) . Sequence analysis from the CXM(R) transformants also implicated FtsI; in this case, the substitutions were V511A and R517H . To verify S357N substitution, the protein sequence of H . influenzae FtsI was threaded through the S . pneumoniae PBP 2X structure giving an average root mean square deviation of the alpha-carbon chains of 0.5 A . The S357N substitution alters both the residue size and charge . One explanation for the contribution of S357N to CXM(R) is that the asparagine side-chain produces unfavourable steric hindrance with the side chain of Val-362 changing the torsion angles of the asparagine residue, which in turn may influence the position of the loop V362-P366 adjacent to the active site . Whilst other groups have examined the contribution of H . influenzae PBPs in ampicillin resistance, this is the first report analysing their role in CXM(R).

Vaccine, 2003 Mar 28, 21(13-14), 1310 - 6
Combined hepatitis B vaccines; FitzSimons D et al.; The status and likely impact of existing and potential new combined hepatitis B vaccines were broadly considered at the Viral Hepatitis Prevention Board (VHPB) meeting in Malta, October 2001 . The currently available and/or licensed combined hepatitis B vaccines in Europe and the prospects for further such vaccines were reviewed . Data on the safety, immunogenicity, and European licensing status and availability of haxavalent vaccines combining hepatitis B (HepB), Haemophilus influenza type b (Hib), diphtheria, tetanus, and pertussis (acellular) (DTPa), and inactivated poliovirus (IPV) antigens were presented . Finally, the impact of the availability of combined hepatitis B vaccines on hepatitis B immunisation programmes in Europe were examined and the added value of combined hepatitis B vaccines globally was estimated.

Cell Microbiol, 2003 Mar, 5(3), 175 - 86
The Haemophilus influenzae Hap autotransporter mediates microcolony formation and adherence to epithelial cells and extracellular matrix via binding regions in the C-terminal end of the passenger domain; Fink DL et al.; The pathogenesis of non-typable Haemophilus influenzae disease begins with colonization of the nasopharynx and is facilitated by bacterial adherence to respiratory mucosa . The H . influenzae Hap autotransporter is a non-pilus adhesin that promotes adherence to epithelial cells and selected extracellular matrix proteins and mediates bacterial aggregation and microcolony formation . In addition, Hap has serine protease activity . Hap contains a 110 kDa internal passenger domain called HapS and a 45 kDa C-terminal translocator domain called Hapbeta . In the present study, we sought to define the structural basis for Hap adhesive activities . Based on experiments using a panel of monoclonal antibodies against HapS, a deletion derivative lacking most of HapS and a purified fragment of HapS, we established that adherence to epithelial cells is mediated by sequences within the C-terminal 311 residues of HapS . In additional experiments, we discovered that bacterial aggregation is also mediated by sequences within the C-terminal 311 residues of HapS and occurs via HapS-HapS interaction between molecules on neighbouring organisms . Finally, we found that adherence to fibronectin, laminin and collagen IV is mediated in part by sequences within the C-terminal 311 residues of HapS and in full by sequences within the C-terminal 511 residues of HapS . Taken together, these results demonstrate that all Hap adhesive activities reside in the C-terminal portion of HapS . Coupled with earlier observations, the current results establish that HapS adhesive activities and HapS protease activity are contained in separate modules of the protein.

Rev Med Chil, 2002 Dec, 130(12), 1373 - 82
{Community-acquired pneumonia in hospitalized adult patients . Clinical presentation and prognostic factors}; Saldias F et al.; BACKGROUND: Community-acquired pneumonia (CAP) is a serious health problem in Chile . AIM: To study prognostic factors on admission and outcome of CAP, in immune competent adult patients, hospitalized in the Catholic University Clinical Hospital . PATIENTS AND METHODS: All adult patients admitted with a CAP in a period of 2 years were prospectively studied . Patients with immunodeficiency, solid tumors or receiving oral adrenal steroids were excluded from the study . RESULTS: In the study period, 463 patients (69 +/- 19 years, 55% male) were evaluated . Ninety four percent were treated with 2nd or 3rd generation cephalosporins . Mean hospital length of stay was 10 days . Mortality during hospital stay was 8% and in the ensuing 30 days, it was 12% . Bacterial etiology was established in 25% of cases . The most frequent pathogens isolated were Streptococcus pneumoniae (10.2%), Haemophilus influenzae (3.7%), Staphylococcus aureus (2.8%) and Gram negative bacilli (5.2%) . Admission prognostic factors associated with hospital mortality were an age over 65 years, presence of comorbidity, chronic neurological and hepatic disease, suspicion of aspiration, duration of symptoms for less than 3 days, presence of dyspnea and altered mental status, absence of cough, fever and chills; low blood pressure, tachypnea, metabolic acidosis, hypoxemia, high blood urea nitrogen, hypernatremia, hyperkalemia, hyperphosphatemia, hypoalbuminemia, multilobar radiographic pulmonary infiltrates, bacteremia, high risk categories of the Fine Index (IV and V), and admission to Intermediate Care Unit or ICU . CONCLUSIONS: The features of community acquired pneumonia of these patients are similar to those reported abroad.

Eur Respir J, 2003 Feb, 21(2), 294 - 302
Community-acquired pneumonia in the elderly: Spanish multicentre study; Zalacain R et al.; Community-acquired pneumonia (CAP) in the elderly has increased as a consequence of an overall increase of the elderly population . A controversy about the aetiology and outcome of CAP in this population still exists and more epidemiological studies are needed . A prospective, 12-month, multicentre study was carried out to assess the clinical characteristics, aetiology, evolution and prognostic factors of elderly patients (> or = 65 yrs) admitted to hospital for CAP . The study included 503 patients (age 76 +/- 7 yrs) . The clinical picture lasted < or = 5 days in 318 (63%) and the main clinical features were cough (n = 407, 81%) and fever (n = 380, 76%) . Aetiological diagnosis was achieved in 199 (40%) cases, with a definite diagnosis obtained in 164 (33%) . Of the 223 microorganisms isolated the main agents found were Streptococcus pneumoniae in 98 (49%) and Haemophilus influenzae in 27 (14%) . A total of 53 patients died (11%) and the multivariate analysis showed the following factors of bad prognosis: previous bed confinement, alteration in mental status, absence of chills, plasma creatinine > or = 1.4 mg x dL(-1), oxygen tension in arterial blood/inspiratorv oxygen fraction ratio < 200 at the time of admission, and shock and renal failure during the evolution . The results of this study may aid in the management of empiric antibiotic treatment in elderly patients with community-acquired pneumonia and the patients who have a greater probability of bad evolution may be identified based on the risk factors.

J Biol Chem, 2003 May 9, 278(19), 16658 - 66 Epub 2003 Feb 26.
Purification and characterization of a chimeric enzyme from Haemophilus influenzae Rd that exhibits glutathione-dependent peroxidase activity; Pauwels F et al.; While belonging to the same family of antioxidant enzymes, members of the peroxiredoxins do not necessarily employ one and the same method for their reduction . Most representatives become reduced with the aid of thioredoxin, whereas some members use AhpF, tryparedoxin, or cyclophilin A . Recent research on a new peroxiredoxin isoform (type C) from Populus trichocarpa has shown that these particular types may also use glutaredoxin instead of thioredoxin . This finding is supported by the occurrence of chimeric proteins composed of a peroxiredoxin and glutaredoxin region . A gene encoding such a fusion protein is enclosed in the Haemophilus influenzae Rd genome . We expressed the H . influenzae protein, denoted here as PGdx, in Escherichia coli and purified the recombinant enzyme . In vitro assays demonstrate that PGdx, in the presence of dithiothreitol or glutathione, is able to protect supercoiled DNA against the metal ion-catalyzed oxidation-system . Enzymatic assays did, indeed, characterize PGdx as a peroxidase, requiring the glutathione redox cycle for the reduction of hydrogen peroxide (k(cat)/K(m) 5.01 x 10(6) s(-1) m(-1)) as well as the small organic hydroperoxide tert-butylhydroperoxide (k(cat)/K(m) 5.67 x 10(4) s(-1) m(-1)) . Enzymatic activity as function of the glutathione concentration deviated from normal Michaelis-Menten kinetics, giving a sigmoidal pattern with an apparent Hill coefficient of 2.9 . Besides the formation of a disulfide-linked PGdx dimer, it was also shown by mass spectrometric analysis that cysteine 49, which is equivalent to the active site cysteine of the peroxiredoxins, undergoes glutathionylation during purification under nonreducing conditions . Based on these results, we propose a model for the catalytic mechanism.

Antimicrob Agents Chemother, 2003 Mar, 47(3), 1017 - 22
Effects of an efflux mechanism and ribosomal mutations on macrolide susceptibility of Haemophilus influenzae clinical isolates; Peric M et al.; This study investigated macrolide resistance mechanisms in clinical Haemophilus influenzae strains with different levels of susceptibility to macrolides . A total of 6,382 isolates were collected during the Alexander Project from 1997 to 2000 . For 96.9% of these isolates, the azithromycin MICs were 0.25 to 4 micro g/ml, and these were defined as baseline strains . For 1.8% of the isolates, the azithromycin MICs were lower (<0.25 micro g/ml), and for 1.3% of the isolates, the MICs were higher (>4 micro g/ml) . These isolates were defined as hypersusceptible and high-level macrolide-resistant strains, respectively . To identify the mechanisms associated with these three susceptibility patterns, representative strains were studied for the presence of macrolide efflux pumps and for ribosomal alterations . Macrolide efflux was studied by measuring the accumulation of radioactive azithromycin and clarithromycin in the presence or absence of carbonyl cyanide m-chlorophenylhydrazone (CCCP), a protonophore . Treatment with CCCP increased the accumulation of macrolides in baseline as well as high-level resistant strains, demonstrating the presence of an efflux mechanism, but not in the 20 hypersusceptible strains tested . Among the 31 strains studied that showed high-level resistance to both azithromycin and clarithromycin, 28 had ribosomal alterations, 7 had mutations in ribosomal protein L4, 11 had mutations in L22, 2 had mutations in 23S rRNA, 8 had multiple mutations, and 3 had no mutations . From these results, we conclude that the vast majority (>98%) of H . influenzae strains have a macrolide efflux mechanism, with a few of these being hyperresistant (1.3%) due to one or several ribosomal mutations . Occasional hypersusceptible strains (1.8%) were found and had no macrolide resistance mechanisms and appeared to be the only truly macrolide-susceptible variants of H . influenzae.

Emerg Infect Dis, 2003 Feb, 9(2), 258 - 61
Invasive type e Haemophilus influenzae disease in Italy; Cerquetti M et al.; We describe the first reported cases of invasive type e Haemophilus influenzae disease in Italy . All five cases occurred in adults . The isolates were susceptible to ampicillin and eight other antimicrobial agents . Molecular analysis showed two distinct type e strains circulating in Italy, both containing a single copy of the capsulation locus.

Protein Eng, 2002 Dec, 15(12), 1005 - 14
Structural basis of ICF-causing mutations in the methyltransferase domain of DNMT3B; Lappalainen I et al.; Mutations in the gene encoding for a de novo methyltransferase, DNMT3B, lead to an autosomal recessive Immunodeficiency, Centromeric instability and Facial anomalies (ICF) syndrome . To analyse the protein structure and consequences of ICF-causing mutations, we modelled the structure of the DNMT3B methyltransferase domain based on Haemophilus haemolyticus protein in complex with the cofactor AdoMet and the target DNA sequence . The structural model has a two-subdomain fold where the DNA-binding region is situated between the subdomains on a surface cleft having positive electrostatic potential . The smaller subdomains of the methyltransferases differ in length and sequences and therefore only the target recognition domain loop was modelled to show the location of an ICF-causing mutation . Based on the model, the DNMT3B recognizes the GC sequence and flips the cytosine from the double-stranded DNA to the catalytic pocket . The amino acids in the cofactor and target cytosine binding sites and also the electrostatic properties of the binding pockets are conserved . In addition, a registry of all known ICF-causing mutations, DNMT3Bbase, was constructed . The structural principles of the pathogenic mutations based on the modelled structure and the analysis of chi angle rotation changes of mutated side chains are discussed.

Pediatr Surg Int, 2002 Dec, 18(8), 728 - 9 Epub 2002 Sep 14.
Primary peritonitis in a child caused by Haemophilus parainfluenzae; Mayer MP et al.; A case of primary peritonitis caused by Haemophilus parainfluenzae in a previously healthy child is presented . To our knowledge, this is the first such documented case.

Arch Dis Child, 2003 Mar, 88(3), 206 - 10
Hib vaccination in infants born prematurely; Heath PT et al.; AIMS: To document the immunogenicity and persistence of antibody to polyribosyl-ribitol phosphate (PRP) as well as the clinical protection against invasive Haemophilus influenzae type b (Hib) disease in premature infants immunised at the routine schedule . METHODS: Blood was obtained at 2, 5, 12, and 64 months of age from a cohort of prematurely born infants (<or=32 weeks gestation) . Anti-PRP antibody concentrations were compared with those of a control cohort of infants born at full term and vaccinated at the same schedule . Hib vaccine failures occurring between October 1992 and October 2000 were reported by paediatricians through an active, prospective, national survey in the UK and Republic of Ireland . The number of prematurely born children with vaccine failure was compared with the corresponding number born at term . RESULTS: Twenty seven prematurely born infants were followed to 5 years of age . Compared with term infants they had a significantly lower geometric mean concentration of anti-PRP antibody and/or a significantly lower proportion above one or both of the conventional protective antibody concentrations (0.15 and 1.0 micro g/ml) at all ages . A total of 165 cases of invasive Hib disease were identified over eight years of national surveillance . Eighteen were premature (<37 weeks); approximately 12 would be expected . The relative risk of UK premature infants developing disease compared with term infants was 1.5 (95% CI 0.9 to 2.6) . CONCLUSIONS: Premature infants develop lower antibody concentrations than term infants following Hib conjugate vaccination . Premature infants may also have an increased risk of clinical vaccine failure, but interpretation is limited by the small number of premature infants developing invasive Hib disease over eight years of national surveillance . Overall, vaccination with Hib conjugate vaccines affords a high level of protection to premature babies.

Bull Soc Pathol Exot, 2002 Nov, 95(4), 257 - 61
{Immunodepression and pulmonary infections}; Yao NA et al.; The acquired immunosuppressed states are increasingly numerous . Pneumopathies are a frequent, serious complication and etiologic diagnosis is often difficult . The nature of the micro-organism in question is a function of the immunizing type of deficiency . In neutropenias, the infections are primarily bacterial, their potential gravity being correlated with the depth of the deficiency into polynuclear, or fungic, especially in prolonged neutropenias . The aspleened states are responsible for a deficit of the macrophage system and contribute to the infections with encapsulated germs (pneumococci, klebsiellas...) . The organic grafts imply an attack of cell-mediated immunity, in the particular case of the auxiliary T lymphocytes (CD4)), with a special predisposition for viral and fungic infections . During VIH infection, the immunizing deficit of CD4 lymphocytes worsens with time . At the early stage, the infections are especially bacterial . At the more advanced stages, the pulmonary pneumocystosis and tuberculosis dominate . At the late stage, finally, deep immunosuppression allows emerging of the atypical mycobacteries . In the deficiencies of humoral immunity (congenital hypogammaglobulinemias, lymphoid hemopathies B), the germs to be mentioned are the pneumococcus, Haemophilus influenzae, the salmonellas and the legionellas . Immunosuppressed pneumopathies are characterized by radio-clinical pictures of very variable gravity, ranging from focused acute pneumopathy to bilateral diffuse pneumopathy with acute respiratory distress syndrome, with phases of atypical tables with respiratory symptomatology larval or absent . The highlighting of the micro-organisms in question requires urgent complementary investigations: hemocultures, bronchiolo-alveolar washing . In certain cases, it will be possible to resort to the transtracheal puncture or transthoracic puncture guided by tomodensitometry, and if necessary to pulmonary biopsy under videothoracoscopy . Emergency of the anti-infectious treatment imposes, in general, a presumptive treatment directed according to the immunizing deficiency in question and etiologic suspicion . It will be associated, if necessary, with urgent measurements of respiratory intensive care.

Proteins, 2003 Apr 1, 51(1), 56 - 67
Structure of the YibK methyltransferase from Haemophilus influenzae (HI0766): a cofactor bound at a site formed by a knot; Lim K et al.; The crystal structures of YibK from Haemophilus influenzae (HI0766) have been determined with and without bound cofactor product S-adenosylhomocysteine (AdoHcy) at 1.7 and 2.0 A resolution, respectively . The molecule adopts an alpha/beta fold, with a topology that differs from that of the classical methyltransferases . Most notably, HI0766 contains a striking knot that forms the binding crevice for the cofactor . The knot formation is correlated with an alternative arrangement of the secondary structure units compared with the classical methyltransferases . Two loop regions undergo conformational changes upon AdoHcy binding . In contrast to the extended conformation of the cofactor seen in the classical methyltransferase structures, AdoHcy binds to HI0766 in a bent conformation . HI0766 and its close sequence relatives are all shorter versions of the more remotely related rRNA/tRNA methyltransferases of the spoU sequence family . We propose that the spoU sequence family contains the same core domain for cofactor binding as HI0766 but has an additional domain for substrate binding . The substrate-binding domain is absent in HI0766 sequence family and may be provided by another Haemophilus influenzae partner protein, which is yet to be identified .

Infect Immun, 2003 Mar, 71(3), 1098 - 108
Position-based scanning for comparative genomics and identification of genetic islands in Haemophilus influenzae type b; Bergman NH et al.; Bacteria exhibit extensive genetic heterogeneity within species . In many cases, these differences account for virulence properties unique to specific strains . Several such loci have been discovered in the genome of the type b serotype of Haemophilus influenzae, a human pathogen able to cause meningitis, pneumonia, and septicemia . Here we report application of a PCR-based scanning procedure to compare the genome of a virulent type b (Hib) strain with that of the laboratory-passaged Rd KW20 strain for which a complete genome sequence is available . We have identified seven DNA segments or H . influenzae genetic islands (HiGIs) present in the type b genome and absent from the Rd genome . These segments vary in size and content and show signs of horizontal gene transfer in that their percent G+C content differs from that of the rest of the H . influenzae genome, they contain genes similar to those found on phages or other mobile elements, or they are flanked by DNA repeats . Several of these loci represent potential pathogenicity islands, because they contain genes likely to mediate interactions with the host . These newly identified genetic islands provide areas of investigation into both the evolution and pathogenesis of H . influenzae . In addition, the genome scanning approach developed to identify these islands provides a rapid means to compare the genomes of phenotypically diverse bacterial strains once the genome sequence of one representative strain has been determined.

J Bacteriol, 2003 Mar, 185(5), 1608 - 15
Chromosomal expression of the Haemophilus influenzae Hap autotransporter allows fine-tuned regulation of adhesive potential via inhibition of intermolecular autoproteolysis; Fink DL et al.; The Haemophilus influenzae Hap autotransporter is a nonpilus adhesin that promotes adherence to respiratory epithelial cells and selected extracellular matrix proteins and facilitates bacterial aggregation and microcolony formation . Hap consists of a 45-kDa outer membrane translocator domain called Hap(beta) and a 110-kDa extracellular passenger domain called Hap(S) . All adhesive activity resides within Hap(S), which also contains protease activity and directs its own secretion from the bacterial cell surface via intermolecular autoproteolysis . In the present study, we sought to determine the relationship between the magnitude of Hap expression, the efficiency of Hap autoproteolysis, and the level of Hap-mediated adherence and aggregation . We found that a minimum threshold of Hap precursor was required for autoproteolysis and that this threshold approximated expression of Hap from a chromosomal allele, as occurs in H . influenzae clinical isolates . Chromosomal expression of wild-type Hap was sufficient to promote significant adherence to epithelial cells and extracellular matrix proteins, and adherence was enhanced substantially by inhibition of autoproteolysis . In contrast, chromosomal expression of Hap was sufficient to promote bacterial aggregation only when autoproteolysis was inhibited, indicating that the threshold for Hap-mediated aggregation is above the threshold for autoproteolysis . These results highlight the critical role of autoproteolysis and an intermolecular mechanism of cleavage in controlling the diverse adhesive activities of Hap.

J Bacteriol, 2003 Mar, 185(5), 1572 - 81
Exogenous glutathione completes the defense against oxidative stress in Haemophilus influenzae; Vergauwen B et al.; Since they are equipped with several strategies by which they evade the antimicrobial defense of host macrophages, it is surprising that members of the genus Haemophilus appear to be deficient in common antioxidant systems that are well established to protect prokaryotes against oxidative stress . Among others, no genetic evidence for glutathione (gamma-Glu-Cys-Gly) (GSH) biosynthesis or for alkyl hydroperoxide reduction (e.g., the Ahp system characteristic or enteric bacteria) is apparent from the Haemophilus influenzae Rd genome sequence, suggesting that the organism relies on alternative systems to maintain redox homeostasis or to reduce small alkyl hydroperoxides . In this report we address this apparent paradox for the nontypeable H . influenzae type strain NCTC 8143 . Instead of biosynthesis, we could show that this strain acquires GSH by importing the thiol tripeptide from the growth medium . Although such GSH accumulation had no effect on growth rates, the presence of cellular GSH protected against methylglyoxal, tert-butyl hydroperoxide (t-BuOOH), and S-nitrosoglutathione toxicity and regulated the activity of certain antioxidant enzymes . H . influenzae NCTC 8143 extracts were shown to contain GSH-dependent peroxidase activity with t-BuOOH as the peroxide substrate . The GSH-mediated protection against t-BuOOH stress is most probably catalyzed by the product of open reading frame HI0572 (Prx/Grx), which we isolated from a genomic DNA fragment that confers wild-type resistance to t-BuOOH toxicity in the Ahp-negative Escherichia coli strain TA4315 and that introduces GSH-dependent alkyl hydroperoxide reductase activity into naturally GSH peroxidase-negative E . coli . Finally, we demonstrated that cysteine is an essential amino acid for growth and that cystine, GSH, glutathione amide, and cysteinylglycine can be catabolized in order to complement cysteine deficiency.

Vet Microbiol, 2003 May 2, 93(1), 79 - 87
Purification and renaturation of membrane neuraminidase from Haemophilus parasuis; Lichtensteiger CA et al.; Haemophilus parasuis, which causes polyserositis, polysynovitis, meningitis, septicemia, and pneumonia in pigs, has emerged as an increasing problem in modern swine production systems . Co-factors for and the pathogenesis of H . parasuis disease are not defined . One of the potential virulence factors of H . parasuis is its neuraminidase (sialidase) . While purifying the H . parasuis neuraminidase from the membrane fraction, we developed a protocol to renature enzymatic activity after enzyme preparations were resolved electrophorectically in denaturing polyacrylamide gels . The H . parasuis neuraminidase co-resolved with recombinant neuraminidase of Vibrio cholera; thus its apparent molecular mass is 82 kilodalton (kDa) . The H . parasuis neuraminidase was associated with the membrane fraction and the purification protocol removed over 99% of the H . parasuis cell protein while retaining over 90% of the neuraminidase activity . Purified protein will provide another avenue to clone the neuraminidase gene that has been refractory to cloning and the protocol will be a means to purify recombinant protein .

Rev Neurol, 2003 Jan 16-31, 36(2), 184 - 94
{Prevention of mental retardation}; Calderon-Gonzalez R et al.; OBJECTIVE: To present within the general field of the conditions causing mental retardation, the preventive strategies for specific application available at the present time . DEVELOPMENT: In spite of the fact that in the majority of cases of mental retardation the etiology is unknown, and for that reason, in them it is not possible to establish preventive strategies, within the last three decades, important research advances have helped to prevent thousands of cases of mental retardation of illnesses caused by Haemophilus influenzae B, measles encephalitis, Rh disease and severe jaundice in newborn infants, congenital hypothyroidism, phenylketonuria and congenital rubella; as well as removing lead from the environment, intervention programs for the proper use of seat belts, child safety seats, and motorcycle and bicycle helmets; early and adequate prenatal care, dietary supplementation with folic acid beginning before conception to reduce the risk of neural tube defects, avoidance of toxic substances during pregnancy like alcohol, and the use of newborn screening tests . CONCLUSION: The primary and secondary prevention of conditions that cause mental retardation continue being a challenge . Require of a review of the present strategies, that frequently inform about the problem, but are not practice in an every day bases (ej . intake of alcohol during pregnancy, the universal use of seat belt and child safety seats during automobile travel) . In the future we may have the possibility of prenatal gene therapy.

Clin Microbiol Infect, 2003 Feb, 9(2), 79 - 85
Carbohydrate-protein conjugate vaccines; Ada G et al.; Various pathogenic bacteria have coats of polysaccharide, many with repeating epitopes . Though polysaccharide vaccines have been available for some time, they induce mainly IgM production, and are only moderately protective in adults and ineffective in young children . It was originally shown in 1931 that the immunogenicity of polysaccharides could be enhanced by conjugating to a protein . The last two decades have witnessed the production and clinical testing of polysaccharide-protein conjugates specific for at least four different bacteria which normally cause considerable mortality and morbidity, especially in young children . In some cases, immunizing children from 4 months of age, with a booster early in the second year, has resulted in remarkably high success rates in protecting them from disease . For one pathogen, Haemophilus influenza type b, the success rate has been sufficiently high (> 95%) to suggest that this disease might, in time, be globally controlled in this way . The results of immunization with conjugate vaccines to Streptococcus pneumoniae, Neisseria meningiditis and Salmonella typhi are also very encouraging . More conjugate preparations are under development.

Rev Esp Quimioter, 2002 Dec, 15(4), 325 - 34
{In vitro activity of moxifloxacin against respiratory pathogens in Latin America}; Lopez H et al.; We compared the in vitro activity of moxifloxacin, levofloxacin and six other antibiotics frequently used in respiratory tract infections, against 1563 Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Haemophilus influenzae and Moraxella catarrhalis clinical isolates from 21 centers in 10 Latin American countries from March 2000 to April 2001 . Moxifloxacin was the most active compound against all the species included . Moxifloxacin was 2- to 4-fold more active than levofloxacin against Gram-positive bacteria . This difference was much higher against levofloxacin-resistant S . pneumoniae isolates (MIC: 0.5 mg/l vs 8 mg/l) . The activity of moxifloxacin against H . influenzae and M . catarrhalis was similar to levofloxacin; all the isolates were inhibited at < or = 1 mg/l concentrations of moxifloxacin and levofloxacin.

Enferm Infecc Microbiol Clin, 2003 Mar, 21(3), 131 - 6
{Sensitivity of Haemophilus influenzae isolates in Spain to 17 oral antibiotics}; Aracil B et al.; INTRODUCTION: The objective of this study was to assess the sensitivity of 400 strains of Haemophilus influenzae isolated in 21 Spanish laboratories in 1999 to 17 oral antibiotics.Methods . An agar dilution method in HT medium was used for sensitivity testing; interpretation of the results followed NCCLS criteria . RESULTS: Percentages of isolates susceptible to the antibiotics tested were as follows: ampicillin 59.5%, amoxicillin/clavulanate 99.2%, loracarbef 66.2%, cefprozil 70.2%, cefaclor 76.2%, cefuroxime 95%, ceftibuten 100%, cefpodoxime 100%, cefixime 100%, rifampin 99.8%, tetracycline 98.2%, chloramphenicol 99.2%, nalidixic acid 97.5%, ciprofloxacin 100%, trovafloxacin 100%, clarithromycin 74%, and azithromycin 100% . CONCLUSIONS: Geographic distribution of sensitivity rates was not homogeneous for some antibiotics . Around 24% of strains were betalactamase producers, requiring higher MICs of antibiotics such as loracarbef, cefprozil and cefaclor than non betalactamase producers . Nevertheless MICs of ceftibuten, cefpodoxime and cefixime were similar in both betalactamase producers and non-producers . Five strains (1.25%) were beta -lactamase (2), but resistant to ampicillin (MIC > or = 8 mg/L) and to amoxicillin/clavulanic acid (MIC > or = 4/2 mg/L) . Only three strains showed intermediate sensitivity to chloramphenicol . These strains and four others were inhibited with > or = 4 mg/L of tetracycline . Only one strain was resistant to tetracycline (MIC: 64 mg/L) and to rifampin (MIC: 256 mg/L) . All strains were sensitive to azithromycin (MICs < or = 4 mg/L) and all were sensitive to ciprofloxacin and trovafloxacin (MICs < or = 0.5 mg/L) . However, ten strains (2.5%) were resistant to nalidixic acid (MIC > or = 4 mg/L).

Enferm Infecc Microbiol Clin, 2003 Mar, 21(3), 126 - 30
{Haemophilus influenzae: phenotype characteristics of strains isolated in 12 Catalan hospitals over one year}; Latorre C et al.; INTRODUCTION: The purpose of this study was to investigate the antigenic and biochemical characteristics, antibiotic susceptibility, and mechanisms for acquiring resistance of Haemophilus influenzae strains isolated in several Catalan hospitals, to determine the current situation regarding this microorganism in our area . METHODS: Serotype, biotype and betalactamase production, as well as susceptibility to eight antimicrobial agents (ampicillin, cefuroxime, cefotaxime, cefixime, clarithromycin, co-trimoxazole, chloramphenicol and ciprofloxacin) were determined in 497 H . influenzae strains isolated from 1 May 1999 to 30 April 2000 in 12 Catalan hospitals . RESULTS: Among the total, 97.5% of strains were nontypable and 50% of the encapsulated strains were serotype b (all isolated from children under 5 years old) . There was a predominance of biotype II, though no age or pathologic tropism was found among any of the biotypes . Our series confirms the previously reported trend to decreasing betalactamase mediated ampicillin resistance in our area, mainly in strains from pediatric patients . More betalactamase negative ampicillin-resistant strains (BLNAR) were isolated in children than in adults . One ciprofloxacin-resistant strain was detected . CONCLUSIONS: Infections caused by encapsulated H . influenzae strains are infrequent in our area and the relative importance of serotype b is decreasing . Mechanisms for acquiring ampicillin resistance other than betalactamase production are emerging . Surveillance of ciprofloxacin susceptibility is required to predict therapeutic failures with this quinolone.

J Chemother, 2002 Dec, 14(6), 591 - 6
Efficacy of amoxicillin-sulbactam, given twice-a-day, for the treatment of community-acquired pneumonia: a clinical trial based on a pharmacodynamic model; Jasovich A et al.; The present multicenter study reports the results of a clinical trial, designed on the basis of a pharmacodynamic study published previously (Bantar et al., J . Chemother 2000; 12: 223-227) to assess the efficacy of amoxicillin/sulbactam (875 mg/125 mg), given orally twice-a-day for 7 days in the treatment of patients with community-acquired pneumonia (CAP) . Eighty-four evaluable subjects older than 19 years with clinical symptoms and features suggestive of CAP, consulting from June 2000 to March 2002 and meeting the PORT risk class I through III, were enrolled in the study . Mean age (y +/- standard deviation) was 46.7 +/- 16.3 and 62% of the patients had some co-morbidity predisposing for CAP . Several individuals (77.4%) fell into a low-risk class (i.e . PORT I or II) and 22.6% of patients belonged to a moderate-risk class at the start of treatment . Six patients (6.45%) had pneumococcal bacteremia . Streptococcus pneumoniae was the organism most frequently isolated (61.9% of all the patients in whom an etiologic diagnosis was made), followed by Haemophilus influenzae . Clinical success was observed in 97.6% of the patients (confidence interval 95%, 94.3%-100%) . Almost all the individuals with clinical success became afebrile within the first 3 days of therapy . Ten patients (11.8%) reported mild or moderate adverse events (especially diarrhea) possibly related to the antimicrobial therapy, but this did not lead to withdrawal from the trial . The results of this study suggest that amoxicillin/sulbactam (875 mg/125 mg) is an efficacious and well tolerated option for treating patients with CAP belonging to a low-moderate risk class and support the use of a short, oral (7-day) b.i.d . regimen.

J Chemother, 2002 Dec, 14(6), 554 - 61
In vitro activity of thiamphenicol against multiresistant Streptococcus pneumoniae, Haemophilus influenzae and Staphylococcus aureus in Italy; Marchese A et al.; Thiamphenicol is a derivative of chloramphenicol characterized by a spectrum comparable to that of the parent compound against multiresistant pathogens but showing satisfactory tolerability . The in vitro activity of thiamphenicol and of 11 comparative drugs against 397 recently isolated antibiotic-resistant and/or invasive pneumococci and 52 multiply-resistant MRSA including 2 VISA strains was determined . Bactericidal activity against Haemophilus influenzae and the post-antibiotic effect on Streptococcus pneumoniae, H . influenzae, Staphylococcus aureus and Escherichia coli were also assessed . Against invasive pneumococci, thiamphenicol and chloramphenicol were the most potent non-beta-lactam molecules together with vancomycin and rifampin . Against high-level penicillin-resistant strains phenicol activities were superior to those of cefotaxime, ceftriaxone and imipenem . Against MRSA thiamphenicol and chloramphenicol were second only to the glycopetides and also inhibited the VISA strains . Thiamphenicol showed a significant PAE (0.33 to 2.9h) on all pathogens studied and a powerful bactericidal effect against beta-lactamase-positive and -negative H . influenzae . These results indicate a good in vitro activity of thiamphenicol against difficult-to-treat multiply resistant pathogens.

Rev Esp Quimioter, 2002 Jun, 15(2), 148 - 51
{Study of the betalactam resistance of Haemophilus influenzae conferred by the bla(ROB-1) gene}; Molina JM et al.; Given the lack of information on the resistance to betalactams conferred by the bla(ROB-1) gene in our area, we decided to undertake a study to determine if and how often it appears in our isolates . We analyzed the presence of the gene in 177 strains of Haemophilus influenzae and compared the results with those from the biochemical tests (nitrocefin) . Sixty-three strains tested positive in the nitrocefin test (35.6%) . The presence of the bla(ROB-1) gene was detected in seven of the 177 strains (3.9%) . All the strains expressing the gene also tested positive in the nitrocefin test . Of the 63 strains that showed biochemical resistance (positive to nitrocefin), seven were found to be caused by the presence of the bla(ROB-1) gene . An epidemiological follow-up for the presence of this gene in our area is therefore advisable.

Rev Esp Quimioter, 2002 Sep, 15(3), 215 - 23
{Trans-gram gene transfer: the case of beta-lactamases}; Galan JC; Lateral or horizontal gene transfer is a phenomenon that has influenced the evolution of microorganisms . Despite the evolutionary trend toward genetic isolation, lateral transfer seems to take place relatively frequently, bridging the gap between very separate species . The acquisition of foreign genes may have accelerated in recent years because of the increase in the adaptive needs of bacteria, particularly through the use of antibiotics . Transfer of genes encoding beta-lactamases from Gram-positive to Gram-negative bacteria (trans-gram transfer) may be suggested on the basis of sequence analysis . We found that the sequences of the beta-lactamases BRO-1 and ACl-1, from the Gram-negative bacterial organisms Moraxella and Acidaminococcus, respectively are abnormally placed among sequences from Gram-positive beta-lactamases in phylogenetic trees . In both cases, the topology of the enzyme (attached to the cellular membrane), the structure of the signal peptide, and the Shine-Dalgarno region suggest that these enzymes originated from Gram-positive organisms . Results remain inconclusive for Haemophilus ROB-1 beta-lactamase.

Mol Microbiol, 2003 Feb, 47(4), 1101 - 11
Identification and characterization of genomic loci unique to the Brazilian purpuric fever clonal group of H . influenzae biogroup aegyptius: functionality explored using meningococcal homology; Li MS et al.; Brazilian purpuric fever (BPF) is a fulminant septicaemic infection of young children, caused by a clonal group of strains of Haemophilus influenzae biogroup aegyptius (Hae), an organism previously solely associated with conjunctivitis . Their special capacity to invade from the initial site of conjunctival infection is unexplained . A polymerase chain reaction (PCR)-amplified subtractive hybridization technique was used to identify genes specific to the BPF clonal group . A copy of bacteriophage HP1 and 46 further chromosomal loci were identified in the BPF but not in the conjunctivitis strain of Hae . Sixteen were characterized further, and one - encoding an analogue of the Legionella pneumophila epithelial cell entry-enhancing protein EnhC - was investigated in depth . Two genes, bpf001 and bpf002, unique to the BPF clonal group were identified between homologues of HI1276 and HI1277 in a complex locus close to H . influenzae genetic island 1, recently identified in pathogenic H . influenzae type b . Bpf001 encodes a protein homologous to EnhC and to the previously uncharacterized product of the meningococcal gene NMB0419 . Functional studies of bpf001 proving intractable, NMB0419 was chosen as a surrogate for investigation and shown to modulate bacterial interaction with monolayers of human respiratory epithelial cells, promoting invasion, the first stage (for Hae) in the pathogenesis of BPF.

Eur J Biochem, 2003 Feb, 270(4), 610 - 24
Structural diversity in lipopolysaccharide expression in nontypeable Haemophilus influenzae . Identification of L-glycerol-D-manno-heptose in the outer-core region in three clinical isolates; Mansson M et al.; Structural elucidation of the lipopolysaccharide (LPS) from three nontypeable Haemophilus influenzae clinical isolates, 1209, 1207 and 1233 was achieved using NMR spectroscopy and ESI-MS on O-deacylated LPS and core oligosaccharide (OS) material as well as ESI-MS(n) on permethylated dephosphorylated OS . It was found that the organisms expressed a tremendous heterogeneous glycoform mixture resulting from the variable length of the OS chains attached to the common structural element of H . influenzae, L-alpha-D-Hepp-(1-->2)-{PEtn-->6}-L-alpha-D-Hepp-(1-->3)-{beta-D-Glcp-(1-->4)}-L-alpha-D-Hepp-(1-->5)-{PPEtn-->4}-alpha-Kdop-(2-->6)-Lipid A . Notably, the O-6 position of the beta-D-Glcp residue could either be occupied by PCho or L-glycero-D-manno-heptose (L,D-Hep), which is a location for L,D-Hep that has not been seen previously in H . influenzae LPS . The outer-core L,D-Hep residue was further chain elongated at the O-6 position by the structural element beta-D-GalpNAc-(1-->3)-alpha-D-Galp-(1-->4)-beta-D-Galp, or sequentially truncated versions thereof . The distal heptose residue in the inner-core was found to be chain elongated at O-2 by the globotetraose unit, beta-D-GalpNAc-(1-->3)-alpha-D-Galp-(1-->4)-beta-D-Galp-(1-->4)-beta-D-Glcp, or sequentially truncated versions thereof . Investigation of LPS from an lpsA mutant of isolate 1233 and a lic1 mutant of isolate 1209 was also performed, which aside from confirming the functions of the gene products, simplified elucidation of the OS extending from the proximal heptose (the lpsA mutant), and showed that the organism exclusively expresses LPS glycoforms comprising the outer-core l,d-Hep residue when PCho is not expressed (the lic1 mutant).

Sex Transm Infect, 2003 Feb, 79(1), 68 - 71
Chancroid: clinical manifestations, diagnosis, and management; Lewis DA; Chancroid is a sexually transmitted disease (STD) caused by the Gram negative bacterium Haemophilus ducreyi and is characterised by necrotising genital ulceration which may be accompanied by inguinal lymphadenitis or bubo formation . H ducreyi is a fastidious organism which is difficult to culture from genital ulcer material . DNA amplification techniques have shown improved diagnostic sensitivity but are only performed in a few laboratories . The management of chancroid in the tropics tends to be undertaken in the context of syndromic management of genital ulcer disease and treatment is usually with erythromycin . A number of single dose regimens are also available to treat H ducreyi infection . Genital ulceration as a syndrome has been associated with increased transmission of human immunodeficiency virus (HIV) infection in several cross sectional and longitudinal studies . Effective and early treatment of genital ulceration is therefore an important part of any strategy to control the spread of HIV infection in tropical countries.

J Clin Microbiol, 2003 Feb, 41(2), 880 - 2
Value of indirect hemagglutination and coagglutination tests for serotyping Haemophilus parasuis; Del Rio ML et al.; An indirect hemagglutination test (IHA) and a coagglutination test (CA) were evaluated using saline, boiled, and autoclaved extracts for serotyping Haemophilus parasuis . CA showed several cross-reactions, whereas IHA gave rise to specific reactions, with minor exceptions . IHA was further compared with the immunodiffusion test (the "gold standard") for the serotyping of 67 field isolates . As a conclusion, IHA is recommended as a useful method for sensitive and specific serotyping of H . parasuis.

Arzneimittelforschung, 2002, 52(12), 903 - 13
Synthesis, antibacterial activity, and toxicity of 7-(isoindolin-5-yl)-4-oxoquinoline-3-carboxylic acids . Discovery of the novel des-F(6)-quinolone antibacterial agent garenoxacin (T-3811 or BMS-284756); Hayashi K et al.; The palladium-catalyzed cross-coupling reaction of 5-(tributylstannyl)isoindoline and its 1- and 3-methyl derivatives with 6-fluoro or 6-unsubstituted 7-bromo-1-cyclopropyl-8-methoxy (or difluoromethoxy)-4-oxoquinoline-3-carboxylate afforded the corresponding 1-cyclopropyl-7-(5-isoindolinyl)-4-oxoquinoline-3- carboxylic acids: 6-fluoro, 1a-7a and 6-nonfluoro, 1b-7b . The in vitro antibacterial spectra of the newly synthesized quinolones were mostly characterized by excellent Gram-positive activity against Staphylococcus aureus and Streptococcus pneumoniae including quinolone-resistant strains, and also by significant Gram-negative activity comparable to 7-(1-piperazinyl)fluoroquinolones . Comparative examinations of the in vitro antibacterial profiles and the in vivo toxicity in terms of intravenous lethality, micronuclei-inducing potential and convulsive activity provided 6-nonfluorinated 1-cyclopropyl-8-(difluoromethoxy)-7-(1-methylisoindolin-5-yl)-4- oxoquinoline-3-carboxylic acid {(+/-)-5b} as the candidate for evaluation of the stereoisomers . The enantiomers (R)-5b and (S)-5b were synthesized via the Suzuki coupling reaction of (R)- and (S)-1-methyl derivatives of 2-(triphenylmethyl)isoindolin-5-boronic acid with the corresponding 7-bromo-8-(difluoromethoxy)-4- oxoquinoline-3-carboxylate . The (R)-5b stereoisomer proved to be 2- to 4-fold more active than the (S)-5b stereoisomer against the organisms tested, with the exception of an equal potency observed with S . pneumoniae IID553 and Haemophilus influenzae ATCC49247 . A noticeable in vitro antibacterial profile of (R)-5b was that it is 16- and 64-fold more active than levofloxacin (CAS 100986-85-4) and ciprofloxacin (CAS 86393-32-0), respectively, against Mycoplasma pneumoniae IID813 (MIC of 0.0313 microgram/ml), and 4-fold more active than ciprofloxacin and levofloxacin against Mycobacterium tuberculosis M-4 (MIC of 0.0313 microgram/ml) . Additional studies indicate that (R)-5b (T-3811, CAS 194804-75-6) exhibits excellent antibacterial activity against a wide range of organisms including anaerobes and common respiratory pathogens, while demonstrating a high selectivity against the mammalian homolog topoisomerases . The methane-sulfonate of (R)-5b (T-3811ME, CAS 223652-90-2) is now undergoing clinical testings.

Jpn J Antibiot, 2000 Jun, 53 Suppl B, 3 - 12
{Etiology of community-acquired pneumonia among adult patients in Japan}; Ishida T; To clarify the etiology of community-acquired pneumonia (CAP) in Japan, the causative pathogens were prospectively investigated in adult patients admitted to Kurashiki Central Hospital . The microbiological diagnosis was based on the results of quantitative sputum culture, blood culture, and other invasive procedures, including transthoracic needle aspiration or bronchoscopic examination . Five hundred fifty-two episodes of CAP in 540 patients were admitted between July 1994 and June 1999 . Causative pathogens were identified in 353 episodes (63.9%) . Several characteristics about the etiology of CAP in Japan were recognized: 1) Streptococcus pneumoniae is the most common pathogen followed by Haemophilus influenzae; 2) Mycoplasma pneumonia is dominant among young patients; 3) Chlomydia pneumoniae is one of the significant pathogens in Japan as well as in western countries; 4) Streptococcus milleri group and anaerobes are important pathogens in patients with suppurative pulmonary diseases; 5) The incidence of Legionella pneumonia is far lower than in western countries; 6) The prevalence of tuberculosis in CAP is still high in Japan; etc . Recognition of these results will lead us to treat patients with prompt antimicrobial therapy.

J Biol Chem, 2003 Apr 11, 278(15), 13496 - 502 Epub 2003 Feb 04.
A catalytic mechanism for D-Tyr-tRNATyr deacylase based on the crystal structure of Hemophilus influenzae HI0670; Lim K et al.; D-Tyr-tRNA(Tyr) deacylase is an editing enzyme that removes d-tyrosine and other d-amino acids from charged tRNAs, thereby preventing incorrect incorporation of d-amino acids into proteins . A model for the catalytic mechanism of this enzyme is proposed based on the crystal structure of the enzyme from Haemophilus influenzae determined at a 1.64-A resolution . Structural comparison of this dimeric enzyme with the very similar structure of the enzyme from Escherichia coli together with sequence analyses indicate that the active site is located in the dimer interface within a depression that includes an invariant threonine residue, Thr-80 . The active site contains an oxyanion hole formed by the main chain nitrogen atoms of Thr-80 and Phe-79 and the side chain amide group of the invariant Gln-78 . The Michaelis complex between the enzyme and D-Tyr-tRNA was modeled assuming a nucleophilic attack on the carbonyl carbon of D-Tyr by the Thr-80 O(gamma) atom and a role for the oxyanion hole in stabilizing the negatively charged tetrahedral transition states . The model is consistent with all of the available data on substrate specificity . Based on this model, we propose a substrate-assisted acylation/deacylation-catalytic mechanism in which the amino group of the D-Tyr is deprotonated and serves as the general base.

Curr Drug Targets Infect Disord, 2002 Dec, 2(4), 339 - 53
Assisting functional assignment for hypothetical Heamophilus influenzae gene products through structural genomics; Gilliland GL et al.; The three-dimensional structures of Haemophilus influenzae proteins whose biological functions are unknown are being determined as part of a structural genomics project to ask whether structural information can assist in assigning the functions of proteins . The structures of the hypothetical proteins are being used to guide further studies and narrow the field of such studies for ultimately determining protein function . An outline of the structural genomics methodological approach is provided along with summaries of a number of completed and in progress crystallographic and NMR structure determinations . With more than twenty-five structures determined at this point and with many more in various stages of completion, the results are encouraging in that some level of functional understanding can be deduced from experimentally solved structures . In addition to aiding in functional assignment, this effort is identifying a number of possible new targets for drug development.

Clin Exp Allergy, 2002 Nov, 32(11), 1589 - 95
T cell cytokine responses to outer membrane proteins of Haemophilus influenzae and the house dust mite allergens Der p 1 in allergic and non-allergic subjects; Epton MJ et al.; BACKGROUND: Haemophilus influenzae are ubiquitous colonizers of the nasopharynx, Little is known about the T cell cytokine responses to the antigens of these bacteria and whether or not the responses may interact with responses to aeroallergen . OBJECTIVE: To measure the T cell cytokine responses to conserved outer membrane protein antigens of Haemophilus influenzae and to house dust mite allergen of subjects allergic to the house dust mite and of subjects without allergic sensitization . METHODS: T cell responses were measured by in vitro proliferation and cytokine release from peripheral blood monocytes (PBMC) . The allergen used was Der p 1 and outer membrane proteins were recombinant polypeptides representing the OMP6 and D15 antigens . RESULTS: The PBMC of most subjects had proliferative responses to OMP6 and D15, which were highly correlated . The pattern of cytokine release was Th1 biased with high levels of IFN-gamma and usually little IL-5 or IL-13 although PBMC from a few subjects did release IL-5 independent of allergic status . IL-10 release was readily detected . There was no difference in the anti-OMP cytokine response of PBMC from subjects without any known allergy and the responses of PBMC from subjects who were highly allergic to house dust mite . The responses to the Der p 1 allergen showed the expected Th2 cytokine release . CONCLUSION: The outer membrane protein antigens of the ubiquitous colonizing bacteria Haemophilus influenzae induce Th1 cytokine responses which are similar for PBMC from non-allergic individuals and subjects with a high degree of allergy to the perennial house dust mite allergen and strong Th2 responses to Der p 1.

Sex Transm Dis, 2003 Feb, 30(2), 114 - 9
Etiology of genital ulcer disease and association with human immunodeficiency virus infection in two tanzanian cities; Ahmed HJ et al.; BACKGROUND: The etiological agent is usually not established in cases of genital ulcer disease (GUD) in Tanzania, since diagnosis and treatment of this disease are based mainly on clinical rather than microbiologic parameters . GUD increases the risk of infection with HIV . However, the association between specific GUD infections and HIV infection has not been fully investigated . GOAL: The goal was to determine the etiology of GUD and the prevalence of HIV infection in patients with GUD in urban areas of Tanzania . STUDY DESIGN: A total of 102 clinical specimens were collected from 52 and 50 patients with GUD in Dar es Salaam and Mbeya, respectively, and from 93 patients with genital discharge in a cross-sectional study . Two polymerase chain reaction (PCR) assays were used to identify either a single target DNA or all three DNAs of the major causes of GUD: Haemophilus ducreyi, Treponema palladum and herpes simplex virus type 2 (HSV-2) . The sera from all patients were tested for antibodies to HIV and T palladum . RESULTS: In Dar es Salaam, DNA from HSV-2, and was detected in 63%, 13%, and 2%, respectively, of the 52 genital ulcer specimens . The corresponding figures in Mbeya were 34%, 10%, and 0% of 50 specimens . Overall, 9% of the 102 patients with GUD were infected with both HSV-2 and, and 39/102 genital ulcer specimens (38%) were negative for the DNA of all three pathogens . The HIV infection rates among GUD patients were 46% and 52% in Dar es Salaam and Mbeya, respectively; among the non-GUD patients, the corresponding rates were 35% and 45%, respectively . The HIV infection rate in Dar es Salaam was significantly higher among women (11/14; 78%) than among men (13/38; 34%) (P = 0.004) . Among the HIV-seropositive GUD patients, 71% and 46% (P < 0.003) were coinfected with HSV-2 in Dar es Salaam and Mbeya, respectively . Furthermore, women with HSV-2 in Dar es Salaam were significantly more likely to be HIV-infected than men (60% versus 39%; P<or= 0.006) . antibodies were detected in 27 (26%) of the 102 GUD patients and 16 (17%) of the 93 non-GUD patients . CONCLUSION: HSV-2 was the most commonly identified agent in the genital ulcer specimens in two urban STD clinics in Tanzania . The prevalence of HIV was high among the STD patients . HSV-2 was detected at significantly higher rates among HIV-seropositive than HIV-seronegative patients with GUD in both Dar es Salaam and Mbeya.

Zhejiang Da Xue Xue Bao Yi Xue Ban, 2002 Feb, 31(1), 47 - 50
{Study on application of PCR in the diagnosis of Haemophilus influenzae pneumonia}; Chen ZM et al.; OBJECTIVE: To study the role of Haemophilus influenzae(Hi) in pneumonia and that of PCR in the diagnosis of Hi pneumonia . METHODS: Hi genus-specific PCR, Hib type-specific PCR and selective Hi culture media were used to detect 83 samples of deep nasal pharyngeal aspiration (NPA), 51 sera from 83 children younger than 3 years with pneumonia and 37 samples of pharyngeal swabs from healthy children . RESULTS: Of 83 NPA samples, 20(24.1 %) were positive by culture, 36 (43.4 %) positive by Hi-PCR and 19 (22.9 %) positive by Hib-PCR.Six out of 51 sera were positive by Hi-PCR and Hib-PCR, but none positive by culture . Of 37 pharyngeal swabs from healthy children, 3 ( 8.1 %) were positive by culture, 6 ( 16.2 %) positive by Hi-PCR and none positive by Hib-PCR . CONCLUSION: Hib-PCR is more appropriate for detecting NPA samples from children with pneumonia because of the high rate of non-typeable Hi carriers in healthy children.

J Antimicrob Chemother, 2002 Dec, 50 Suppl S2, 59 - 73
Dynamics of nasopharyngeal colonization by potential respiratory pathogens; Garcia-Rodriguez JA et al.; Studies have shown that colonization of the nasopharynx by potential respiratory pathogens Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis is established early in childhood, although rates vary greatly according to locality, sampling frequency, individual and social factors . Factors influencing colonization and elimination are not as yet fully understood, but adhesion to mucosal receptors and immune responses are implicated in addition to bacterial properties and colonization resistance dynamics . Colonization in children and adults has been intensively studied in various localities . Potential pathogens are more likely to colonize the nasopharynx of children prone to recurrent otitis media, where impaired local immunity and repeated exposure to respiratory pathogens are additional risk factors . Adults with chronic respiratory tract disease also have higher carriage rates . The factors contributing to increased risk of carriage of potential respiratory pathogens, as well as to clinical infection and antimicrobial resistance, are summarized in this review.

J Antimicrob Chemother, 2002 Dec, 50 Suppl S2, 21 - 6
Susceptibility patterns of bacteria causing community-acquired respiratory infections in Spain: the SAUCE project; Garcia-De-Lomas J et al.; Multicentre surveillance is essential in order to monitor the prevalence of certain resistance phenotypes and to identify rapidly the emergence of new ones . However, many surveillance studies are based either on a relatively small number of isolates from a single country, or on a large number of isolates from many different countries and so are not equally meaningful . Extensive national multicentre surveillance would provide a more reliable strategy for assessing the extent of antimicrobial resistance in individual countries . This article describes Spanish experience with the surveillance network SAUCE, and summarizes the main results on antimicrobial resistance in the three key bacterial pathogens involved in community-acquired respiratory tract infections in Spain: Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes.

Eur J Immunol, 2002 Nov, 32(11), 3225 - 34
Pre-B-cell colony-enhancing factor, whose expression is up-regulated in activated lymphocytes, is a nicotinamide phosphoribosyltransferase, a cytosolic enzyme involved in NAD biosynthesis; Rongvaux A et al.; The murine homologue of the previously identified human "pre-B-cell colony-enhancing factor" (PBEF) gene coding for a putative cytokine has been identified by screening a subtractive library enriched in genes expressed in activated T lymphocytes . Unlike most cytokine genes known to date, the PBEF gene is ubiquitously expressed in lymphoid and non-lymphoid tissues and displays significant homology with genes from primitive metazoans (marine sponges) and prokaryotic organisms . Recently, a bacterial protein encoded by nadV, a gene from the prokaryote Haemophilus ducreyi displaying significant homology with PBEF, has been identified as a nicotinamide phosphoribosyltranferase (NAmPRTase), an enzyme involved in nicotinamide adenine dinucleotide (NAD) biosynthesis . Using a panel of antibodies to murine PBEF, we demonstrate in this work that, similarly to its microbial counterpart, the murine protein is a NAmPRTase, catalyzing the condensation of nicotinamide with 5-phosphoribosyl-1-pyrophosphate to yield nicotinamide mononucleotide, an intermediate in the biosynthesis of NAD . The role of PBEF as a NAmPRTase was further confirmed by showing that the mouse gene was able to confer the ability to grow in the absence of NAD to a NAmPRTase-defective bacterial strain . The present findings are in keeping with the ubiquitous nature of this protein, and indicate that NAD biosynthesis may play an important role in lymphocyte activation.

Int Urol Nephrol, 2002, 34(1), 23 - 4
Haemophilus influenzae acute pyelonephritis in the elderly; Demetrios P et al.; Haemophilus influenzae has rarely been implicated as the causative agent of urinary tract infections in adults . We report a case of H . influenzae pyelonephritis in a 78-year old man with benign prostatic hyperplasia and discuss the possible underestimation of the true incidence of H . influenzae in adult urinary tract infections.

Biophys J, 2003 Feb, 84(2 Pt 1), 794 - 804
Analysis of metabolic capabilities using singular value decomposition of extreme pathway matrices; Price ND et al.; It is now possible to construct genome-scale metabolic networks for particular microorganisms . Extreme pathway analysis is a useful method for analyzing the phenotypic capabilities of these networks . Many extreme pathways are needed to fully describe the functional capabilities of genome-scale metabolic networks, and therefore, a need exists to develop methods to study these large sets of extreme pathways . Singular value decomposition (SVD) of matrices of extreme pathways was used to develop a conceptual framework for the interpretation of large sets of extreme pathways and the steady-state flux solution space they define . The key results of this study were: 1), convex steady-state solution cones describing the potential functions of biochemical networks can be studied using the modes generated by SVD; 2), Helicobacter pylori has a more rigid metabolic network (i.e., a lower dimensional solution space and a more dominant first singular value) than Haemophilus influenzae for the production of amino acids; and 3), SVD allows for direct comparison of different solution cones resulting from the production of different amino acids . SVD was used to identify key network branch points that may identify key control points for regulation . Therefore, SVD of matrices of extreme pathways has proved to be a useful method for analyzing the steady-state solution space of genome-scale metabolic networks.

Trans Am Ophthalmol Soc, 2002, 100, 153 - 9
Vitreous penetration of orally administered gatifloxacin in humans; Hariprasad SM et al.; PURPOSE: To investigate the penetration of gatifloxacin, a novel extended-spectrum fluoroquinolone antibiotic, into the vitreous humor after oral administration . METHODS: A prospective, nonrandomized clinical study of 20 consecutive patients scheduled for pars plana vitrectomy surgery between September 2001 and February 2002 at the Cullen Eye Institute, Houston, Texas . Aqueous, vitreous, and serum samples were obtained and analyzed from 20 patients after oral administration of two 400-mg gatifloxacin tablets taken 12 hours apart before surgery . Assays were performed using high-performance liquid chromatography . RESULTS: Mean gatifloxacin concentrations in serum (n = 19), vitreous (n = 19), and aqueous (n = 10) 4.98 +/- 1.14 micrograms/mL, 1.35 +/- 0.36 microgram/mL, and 1.09 +/- 0.57 micrograms/mL, respectively . Mean sampling times after oral administration of the second gatifloxacin tablet for serum, vitreous, and aqueous were 2.99 +/- 0.73 hours, 3.79 +/- 0.81 hours, and 3.71 +/- 0.87 hours, respectively . The percentages of serum gatifloxacin concentration achieved in the vitreous and aqueous were 27.13% and 21.85%, respectively . Mean inhibitory vitreous and aqueous MIC90 levels were achieved against a wide spectrum of pathogens, including Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Propionibacterium acnes, Haemophilus influenzae, Escherichia coli, Bacillus cereus, Neisseria gonorrhoeae, Proteus mirabilis, and other organisms . CONCLUSIONS: Gatifloxacin is a novel fourth-generation fluoroquinolone antibiotic that has MIC90 levels significantly lower than those of other fluoroquinolone agents . Furthermore, it penetrates well into the vitreous cavity in the noninflamed eye . Potential uses for oral gatifloxacin may include prophylaxis against endophthalmitis in open-globe injuries, surgical prophylaxis against postoperative endophthalmitis, and adjunctive therapy for the current management of bacterial endophthalmitis.

J Craniofac Surg, 2003 Jan, 14(1), 37 - 40
Acute bacterial suppurative parotitis: microbiology and management; Brook I; The parotid gland is the salivary gland most commonly affected by inflammation . The most common pathogens associated with acute bacterial parotitis are Staphylococcus aureus and anaerobic bacteria . The predominant anaerobes include gram-negative bacilli (including pigmented Prevotella and Porphyromonas spp.), Fusobacterium spp., and Peptostreptococcus spp . Streptococcus spp . (including S . pneumoniae) and gram-negative bacilli (including Escherichia coli) have also been reported . Gram-negative organisms are often seen in hospitalized patients . Organisms less frequently found are Arachnia, Haemophilus influenzae, Klebsiella pneumoniae, Salmonella spp., Pseudomonas aeruginosa, Treponema pallidum, cat-scratch bacillus, and Eikenella corrodens . Mycobacterium tuberculosis and atypical mycobacteria are rare causes of parotitis . Therapy includes maintenance of hydration and administration of parenteral antimicrobial therapy . Once an abscess has formed surgical drainage is required . The choice of antimicrobial depends on the etiologic agent . Maintenance of good oral hygiene, adequate hydration, and early and proper therapy of bacterial infection of the oropharynx may reduce the occurrence of suppurative parotitis.

Blood, 2003 May 15, 101(10), 4219 - 21 Epub 2003 Jan 23.
Nephrotic syndrome with crescent formation and massive IgA deposition following allogeneic bone marrow transplantation for natural killer cell leukemia/lymphoma; Kimura S et al.; We describe herein a case of nephrotic syndrome (NS) following allogeneic bone marrow transplantation (allo-BMT) for natural killer cell leukemia/lymphoma . Histologic studies defined the diagnosis as crescentic glomerulonephritis with massive immunoglobulin A (IgA) deposition, which has never been reported in NS cases following allo-BMT . Most of the massive infiltrated cells in the interstice were CD3(+)CD4(-)CD8(+) T cells derived from the donor . We observed mesangial deposition of Haemophilus parainfluenza outer membrane (OMHP) antigen and decreased glycosylation of the IgA1 hinge in the recipient's samples is consistent with the recently reported pathogenesis of IgA nephropathy . Further, the titer of IgA antibody against the donor serum was as high as other IgA nephropathy cases . These findings suggest that NS and crescentic glomerulonephritis in this case occurred as one of the forms of chronic graft-versus-host disease (GVHD), and that IgA deposition was associated with H parainfluenza and decreased glycosylation of the IgA1 hinge.

Protein Sci, 2003 Feb, 12(2), 349 - 60
Structural variation and inhibitor binding in polypeptide deformylase from four different bacterial species; Smith KJ et al.; Polypeptide deformylase (PDF) catalyzes the deformylation of polypeptide chains in bacteria . It is essential for bacterial cell viability and is a potential antibacterial drug target . Here, we report the crystal structures of polypeptide deformylase from four different species of bacteria: Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, and Escherichia coli . Comparison of these four structures reveals significant overall differences between the two Gram-negative species (E . coli and H . influenzae) and the two Gram-positive species (S . pneumoniae and S . aureus) . Despite these differences and low overall sequence identity, the S1' pocket of PDF is well conserved among the four enzymes studied . We also describe the binding of nonpeptidic inhibitor molecules SB-485345, SB-543668, and SB-505684 to both S . pneumoniae and E . coli PDF . Comparison of these structures shows similar binding interactions with both Gram-negative and Gram-positive species . Understanding the similarities and subtle differences in active site structure between species will help to design broad-spectrum polypeptide deformylase inhibitor molecules.

Protein Sci, 2003 Feb, 12(2), 327 - 36
Crystal structure of a trimeric form of dephosphocoenzyme A kinase from Escherichia coli; O'Toole N et al.; Coenzyme A (CoA) is an essential cofactor used in a wide variety of biochemical pathways . The final step in the biosynthesis of CoA is catalyzed by dephosphocoenzyme A kinase (DPCK, E.C . 2.7.1.24) . Here we report the crystal structure of DPCK from Escherichia coli at 1.8 A resolution . This enzyme forms a tightly packed trimer in its crystal state, in contrast to its observed monomeric structure in solution and to the monomeric, homologous DPCK structure from Haemophilus influenzae . We have confirmed the existence of the trimeric form of the enzyme in solution using gel filtration chromatography measurements . Dephospho-CoA kinase is structurally similar to many nucleoside kinases and other P-loop-containing nucleotide triphosphate hydrolases, despite having negligible sequence similarity to these enzymes . Each monomer consists of five parallel beta-strands flanked by alpha-helices, with an ATP-binding site formed by a P-loop motif . Orthologs of the E . coli DPCK sequence exist in a wide range of organisms, including humans . Multiple alignment of orthologous DPCK sequences reveals a set of highly conserved residues in the vicinity of the nucleotide/CoA binding site.

Int J STD AIDS, 2002 Dec, 13 Suppl 2, 38 - 41
Vaccine candidates in STD; Fletcher MA; Sexually transmitted diseases (STDs) are caused by organisms that infect the mucosal surfaces of the genitourinary tract . In spite of its public health importance, current STD vaccine research lags behind work against pathogens that target another mucosal region, the respiratory tract . In the latter case, live-attenuated viral vaccines, killed whole-cell bacterial vaccines, subunit/protein bacterial vaccines, and bacterial polysaccharide vaccines have been enormously successful . To move STD vaccine research forward, complex issues must be resolved . Those include selection of an appropriate antigen (e.g . scientific feasibility and intellectual property rights), the manufacture of the vaccine (e.g . delivery systems, formulation processes, and production steps), and the appropriate public health approach (e.g . medical indications and marketing aspects) . Particular scientific problems have delayed STD vaccine development, like incomplete attenuation (human herpes simplex virus type 2), accentuated immunopathology (Chlamydia trachomatis), poor immunogenicity (Treponema pallidum), and broad antigenic heterogeneity (Neisseria gonorrhoeae) . Nevertheless, efforts continue with the use of protein antigens: for example, the haemolysin toxoid of Haemophilus ducreyi; the major outer membrane protein(s) of N . gonorrhoeae and C . trachomatis; the glycoprotein D of human herpes simplex virus type 2; and the proteins E6 and E7 of human papilloma virus . It may be predicted that eventual STD vaccines (administered either for prophylaxis or for therapy) will use approaches that include (1) live-attenuated viruses, (2) subunit proteins or inactivated whole organisms given with mucosal adjuvants or with cellular immune response adjuvants, and (3) DNA plasmids expressing the vaccine antigen.

Genome Biol . 2002;3(12):RESEARCH0077 . Epub 2002 Nov 26.
JEvTrace: refinement and variations of the evolutionary trace in JAVA; Joachimiak MP et al.; BACKGROUND: Details of functional speciation within gene families can be difficult to identify using standard multiple sequence alignment (MSA) methods . The evolutionary trace (ET) was developed as a visualization tool to combine MSA, phylogenetic and structural data for identification of functional sites in proteins . The method has been successful in extracting evolutionary details of functional surfaces in a number of biological systems and modifications of the method are useful in creating hypotheses about the function of previously unannotated genes . We wish to facilitate the graphical interpretation of disparate data types through the creation of flexible software implementations . RESULTS: We have implemented the ET method in a JAVA graphical interface, JEvTrace . Users can analyze and visualize ET input and output with respect to protein phylogeny, sequence and structure . Function discovery with JEvTrace is demonstrated on two proteins with recently determined crystal structures: YlxR from Streptococcus pneumoniae with a predicted RNA-binding function, and a Haemophilus influenzae protein of unknown function, YbaK . To facilitate analysis and storage of results we propose a MSA coloring data structure . The sequence coloring format readily captures evolutionary, biological, functional and structural features of MSAs . CONCLUSIONS: Protein families and phylogeny represent complex data with statistical outliers and special cases . The JEvTrace implementation of the ET method allows detailed mining and graphical visualization of evolutionary sequence relationships.

Peptides, 2002 Dec, 23(12), 2071 - 83
Development of novel antibacterial peptides that kill resistant isolates; Cudic M et al.; The rapid emergence of bacterial strains that are resistant to current antibiotics requires the development of novel types of antimicrobial compounds . Proline-rich cationic antibacterial peptides such as pyrrhocoricin kill responsive bacteria by binding to the 70 kDa heat shock protein DnaK and inhibiting protein folding . We designed and synthesized multiply protected dimeric analogs of pyrrhocoricin and optimized the in vitro antibacterial efficacy assays for peptide antibiotics . Pyrrhocoricin and the designed dimers killed beta-lactam, tetracycline- or aminoglycoside-resistant strains of Escherichia coli, Salmonella typhimurium, Klebsiella pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis in the submicromolar or low micromolar concentration range . One of the peptides also killed Pseudomonas aeruginosa . The designed dimers showed improved stability in mammalian sera compared to the native analog . In a murine H . influenzae lung infection model, a single dose of a dimeric pyrrhocoricin analog reduced the bacteria in the bronchoalveolar lavage when delivered intranasally . The solid-phase synthesis was optimized for large-scale laboratory preparations.

Mol Microbiol, 2003 Feb, 47(3), 607 - 17
Human milk lactoferrin is a serine protease that cleaves Haemophilus surface proteins at arginine-rich sites; Hendrixson DR et al.; Lactoferrin is a member of the lactotransferrin family of non-haem, iron-binding glycoproteins and is found at high concentrations in all human secretions, where it plays a major role in mucosal defence . In recent work, we observed that lactoferrin has proteolytic activity and attenuates the pathogenic potential of Haemophilus influenzae by cleaving and removing two putative colonization factors, namely the IgA1 protease protein and the Hap adhesin . Experiments with protease inhibitors further suggested that lactoferrin may belong to a serine protease family . In the present study we explored the mechanism of lactoferrin protease activity and discovered that mutation of either Ser259 or Lys73 results in a dramatic decrease in proteolysis . Examination of the crystal structure revealed that these two residues are located in the N-terminal lobe of the protein, adjacent to a 12-15 A cleft that separates the N-lobe and the C-lobe and that can readily accommodate large polypeptide substrates . In additional work, we found that lactoferrin cleaves IgA1 protease at an arginine-rich region defined by amino acids 1379-1386 (RRSRRSVR) and digests Hap at an arginine-rich sequence between amino acids 1016 and 1023 (VRSRRAAR) . Based on our results, we conclude that lactoferrin is a serine protease capable of cleaving arginine-rich sequences . We speculate that Ser259 and Lys73 form a catalytic dyad, reminiscent of a number of bacterial serine proteases . In addition, we speculate that lactoferrin may cleave arginine-rich sequences in a variety of microbial virulence proteins, contributing to its long-recognized antimicrobial properties.

Immunol Cell Biol, 2003 Feb, 81(1), 46 - 51
Bacterial otitis media: current vaccine development strategies; Cripps AW et al.; Otitis media is the most common reason for children less than 5 years of age to visit a medical practitioner . Whilst the disease rarely results in death, there is significant associated morbidity . The most common complication is loss of hearing at a critical stage of the development of speech, language and cognitive abilities in children . The cause and pathogenesis of otitis media is multifactorial . Among the contributing factors, the single most important are viral and bacterial infections . Infection with respiratory syncytial virus, influenza viruses, para-influenza viruses, enteroviruses and adenovirus are most commonly associated with acute and chronic otitis media . Streptococcus pneumoniae, non-typeable Haemophilus influenzae and Moraxella catarrhalis are the most commonly isolated bacteria from the middle ears of children with otitis media . Treatment of otitis media has largely relied on the administration of antimicrobials and surgical intervention . However, attention has recently focused on the development of a vaccine . For a vaccine to be effective against bacterial otitis media, it must, at the very least, contain antigens that induce a protective immune response in the middle ear against the three most common infecting bacteria . Whilst over the past decade there has been significant progress in the development of vaccines against invasive S . pneumoniae disease, these vaccines are less efficacious for otitis media . The search for candidate vaccine antigens for non-typeable H . influenzae are well advanced whilst less progress has been made for M . catarrhalis . No human studies have been conducted for non-typeable H . influenzae or M . catarrhalis and the concept of a tribacterial vaccine remains to be tested in animal models . Only when vaccine antigens are determined and an understanding of the immune responses induced in the middle ear by infection and immunization is gained will the formulation of a tribacterial vaccine against otitis media be possible.

Drugs, 2003, 63(3), 311 - 40
Amoxicillin/clavulanic acid: a review of its use in the management of paediatric patients with acute otitis media; Easton J et al.; Amoxicillin/clavulanic acid (Augmentin), Augmentin ES-600 is a well established, orally administered combination of amoxicillin (a semisynthetic antibacterial agent) and clavulanic acid (a beta-lactamase inhibitor) . Amoxicillin/clavulanic acid shows good activity against the main pathogens associated with acute otitis media (AOM), including penicillin-susceptible and -intermediate strains of Streptococcus pneumoniae, and beta-lactamase producing strains of Haemophilus influenzae and Moraxella catarrhalis . It has moderate activity against penicillin-resistant S . pneumoniae; a high-dose formulation has been developed with the aim of providing better coverage for penicillin-resistant strains . Amoxicillin/clavulanic acid (conventional formulations, mostly 40/10 mg/kg/day in three divided doses) produced clinical response rates similar to those of oral cephalosporin comparators and similar to or significantly greater than those for intramuscular ceftriaxone in randomised trials in paediatric patients with AOM (mean age approximately 2 to 5 years) . Clinical response rates were generally similar for amoxicillin/clavulanic acid and macrolide comparators (mean patient age approximately 1 to 6 years), although significantly better clinical and bacteriological responses were seen versus azithromycin in one randomised trial (mean patient age approximately 1 year) . The high-dose formulation of amoxicillin/clavulanic acid (90/6.4 mg/kg/day in two divided doses) eradicated a high proportion of penicillin-resistant S . pneumoniae (penicillin MICs 2 or 4 mg/L) in a large noncomparative trial in children with AOM (upper limit of the US indication for S . pneumoniae is 2 mg/L) . Amoxicillin/clavulanic acid is generally well tolerated . A low total incidence of adverse events (3.6%) and no serious events were reported from a large paediatric postmarketing study . The most frequently reported adverse events in children are mild gastrointestinal disturbances . Diarrhoea is generally less frequent with twice-daily than with three-times-daily treatment . The new high-dose formulation showed similar tolerability to a conventional twice-daily formulation (45/6.4 mg/kg/day) in a well controlled trial . CONCLUSIONS: Amoxicillin/clavulanic acid is a well established broad-spectrum antibacterial treatment which is effective and well tolerated in the treatment of AOM in paediatric patients . The high-dose combination should prove valuable in treating AOM caused by penicillin-intermediate and -resistant S . pneumoniae (approved in the US for penicillin MIC < or =2 mg/L) . Based on recent recommendations and the available data, high-dose amoxicillin/clavulanic acid can be considered a treatment of choice for recurrent or persistent paediatric AOM (after failure of amoxicillin alone) where involvement of resistant pathogens is suspected.

Tunis Med, 2002 May, 80(5), 286 - 7
{Haemophilus influenzae purulent meningitis in adults: looking for a predisposing factor}; Boukadida J et al.; We bring back an adult case of purulent meningitis to Haemophilus influenzae . We insist on the particular aspects of the host of this meningitis type at the adult . These aspects must be searched every time that Haemophilus influenzae is isolated in cerebrospinal fluid in adult's meningitis.

J Biol Chem, 2003 Mar 28, 278(13), 11513 - 9 Epub 2003 Jan 17.
High resolution structure of an alternate form of the ferric ion binding protein from Haemophilus influenzae; Shouldice SR et al.; The periplasmic iron binding protein of pathogenic Gram-negative bacteria performs an essential role in iron acquisition from transferrin and other iron sources . Structural analysis of this protein from Haemophilus influenzae identified four amino acids that ligand the bound iron: His(9), Glu(57), Tyr(195), and Tyr(196) . A phosphate provides an additional ligand, and the presence of a water molecule is required to complete the octahedral geometry for stable iron binding . We report the 1.14-A resolution crystal structure of the iron-loaded form of the H . influenzae periplasmic ferric ion binding protein (FbpA) mutant H9Q . This protein was produced in the periplasm of Escherichia coli and, after purification and conversion to the apo form, was iron-loaded . H9Q is able to bind ferric iron in an open conformation . A surprising finding in the present high resolution structure is the presence of EDTA located at the previously determined anion ternary binding site, where phosphate is located in the wild type holo and apo structures . EDTA contributes four of the six coordinating ligands for iron, with two Tyr residues, 195 and 196, completing the coordination . This is the first example of a metal binding protein with a bound metal.EDTA complex . The results suggest that FbpA may have the ability to bind and transport iron bound to biological chelators, in addition to bare ferric iron.

Emerg Infect Dis, 2003 Jan, 9(1), 10 - 6
Dead bugs don't mutate: susceptibility issues in the emergence of bacterial resistance; Stratton CW; The global emergence of antibacterial resistance among common and atypical respiratory pathogens in the last decade necessitates the strategic application of antibacterial agents . The use of bactericidal rather than bacteriostatic agents as first-line therapy is recommended because the eradication of microorganisms serves to curtail, although not avoid, the development of bacterial resistance . Bactericidal activity is achieved with specific classes of antimicrobial agents as well as by combination therapy . Newer classes of antibacterial agents, such as the fluoroquinolones and certain members of the macrolide/lincosamine/streptogramin class have increased bactericidal activity compared with traditional agents . More recently, the ketolides (novel, semisynthetic, erythromycin-A derivatives) have demonstrated potent bactericidal activity against key respiratory pathogens, including Streptococcus pneumoniae, Haemophilus influenzae, Chlamydia pneumoniae, and Moraxella catarrhalis . Moreover, the ketolides are associated with a low potential for inducing resistance, making them promising first-line agents for respiratory tract infections.

Jpn J Antibiot, 2002 Oct, 55(5), 537 - 67
{Susceptibilities of bacteria isolated from patients with lower respiratory infectious diseases to antibiotics (2000)}; Shimada K et al.; From October 2000 to September 2001, we collected the specimen from 410 patients with lower respiratory tract infections in 16 institutions in Japan, and investigated the susceptibilities of isolated bacteria to various anti-bacterial agents and antibiotics and patients' characteristics . Of 499 strains that were isolated from specimen (mainly from sputum) and assumed to be bacteria causing in inflammation, 493 strains were investigated . The breakdown of the isolated bacteria were: Staphylococcus aureus 78, Streptococcus pneumoniae 73, Haemophilus infiuenzae 99, Pseudomonas aeruginosa (non-mucoid) 64, P . aeruginosa (mucoid) 14, Klebsiella pneumoniae 25, Moraxella subgenus Branhamella catarrhalis 21, etc . Of 78 S . aureus strains, those with 4 micrograms/ml or more of MIC of oxacillin (methicillin-resistant S . aureus: MRSA) occupied 53.8% . Vancomycin and arbekacin had the most potent activities against MRSA as observed in 1999 . The frequency of S . pneumoniae exhibiting low sensitivity to penicillin (penicillin-intermediate S . pneumoniae: PISP + penicillin-resistant S . pneumoniae: PRSP) was 38.4% being consistent with that in 1999 (34.7%) . PRSP accounted for 11.0% of the total, being more than that in 1999 (3.0%) . Carbapenems had strong activities against S . pneumoniae . Especially, panipenem inhibited the growth of all 73 strains at 0.125 microgram/ml . Generally, all drugs had strong activities against H . influenzae with MIC80s of 8 micrograms/ml or less . The drug that had the strongest activity against H . infiuenzae was levofloxacin, which inhibited the growth of 94 of the 99 strains at 0.063 microgram/ml . Tobramycin had a strong activity against P . aeruginosa (both mucoid and non-mucoid) with MIC80 of 1 microgram/ml . The mucoid strain was little isolated (14 strains) but the susceptibilities to all drugs were better than the non-mucoid strain . K . pneumoniae showed good susceptibilities to all drugs except ampicillin and the MIC80S were 2 micrograms/ml or less . Particularly, cefpirome, cefozopran, and levofloxacin had strong bactericidal activities against K . pneumoniae with MIC80s of 0.125 microgram/ml, and cefotiam, second-generation cephems, also had a favorable activity being MIC80 of 0.25 microgram/ml . Also, all drugs generally had strong activities against M . (B.) catarrhalis . MIC80s of all drugs were 2 micrograms/ml or less . The drug having the strongest activity was imipenem and levofloxacin inhibiting all 21 strains at 0.063 microgram/ml . Most of the patients with respiratory infection were aged 70 years or older, accounting for approximately a half of the total (44.4%) . As for the incidence by the diseases, bacterial pneumonia and chronic bronchitis were the highest, being noted in 38.0% and 31.7% of all the patients, respectively . The bacteria frequently isolated from the patients with bacterial pneumonia were S . aureus (18.3%) and S . pneumoniae (16.1%) . In contrast, H . infiuenzae (20.4%) and P . aeruginosa (both mucoid and non-mucoid: 16.7%) were frequently isolated from the patients with chronic bronchitis . Before the drug administration, the bacteria frequently isolated from all the patients were S . pneumoniae (24.3%) and H . infiuenzae (26.7%) . The frequency of isolated S . pneumoniae tended to decrease with the increase in the number of administration days while that of isolated H . infiuenzae did not . The frequency of isolated P . aeruginosa tended to increase with the duration of administration . The isolated bacteria were comparable between the patients already treated with penicillins and cephems . In the patients treated with aminoglycosides, macrolides, and quinolones, P . aeruginosa was most frequently isolated (33.3 to 40.0%).

Jpn J Antibiot, 2002 Oct, 55(5), 524 - 36
{Antimicrobial susceptibility of Streptococcus pneumoniae and Haemophilus influenzae isolated in major hospitals in Nagano Prefecture}; Yamazaki Y et al.; We determined the minimum inhibitory concentration (MIC) of various antimicrobial agents against 108 strains of Streptococcus pneumoniae and 144 strains of Haemophilus influenzae isolated from respiratory organs in the First Department of Internal Medicine, Shinshu University, and affiliated hospitals between January 2000 and February 2001 . The following results were obtained . 1 . Fifty-one (47.2%), 56 (51.9%), and 1 (0.9%) of 108 strains of S . pneumoniae were classified as penicillin-susceptible S . pneumoniae (PSSP), penicillin-intermediate S . pneumoniae (PISP), and penicillin-resistant S . pneumoniae (PRSP), respectively . 2 . Three carbapenems had potent antimicrobial activity against PISP and PRSP . Furthermore, none of the strains were highly resistant (MIC > 2 micrograms/ml) to benzylpenicillin, ampicillin (ABPC), sulbactam/ampicillin (SBT/ABPC), cefotaxime (CTX), or cefepime (CFPM) . 3 . Eleven (7.6%) and 6 (4.2%) of 144 strains of H . influenzae were classified as beta-lactamase-producing ABPC-resistant strains and beta-lactamase negative ABPC-resistant H . influenzae (BLNAR), respectively . Levofloxacin, sulfamethoxazole/trimethoprim, and meropenem had potent antimicrobial activity against these resistant strains . 4 . BLNAR strains were more highly resistant to CTX, CFPM, SBT/ABPC, and cefaclor than beta-lactamaseproducing strains . 5 . In our surveillance study regarding clinical isolates of S . pneumoniae and H . influenzae from respiratory organs in Nagano prefecture, there were regional differences in the isolation rate and antimicrobial susceptibility . The isolation rates of resistant strains were lower than those reported in a nationwide survey.

Vaccine, 2003 Jan 30, 21(7-8), 716 - 20
Evaluation of the national immunisation programme in the Netherlands: immunity to diphtheria, tetanus, poliomyelitis, measles, mumps, rubella and Haemophilus influenzae type b; de Melker HE et al.; The immunity to vaccine-preventable diseases included in the Dutch immunisation programme in the general population and among orthodox reformed individuals who refuse vaccination was assessed . The programme induces good protection . However, a large proportion of adults lacks diphtheria and tetanus immunity . Measles, mumps and rubella seroprevalence was somewhat lower among vaccinated compared to unvaccinated cohorts . The prevalence of HibPS antibodies declined during 2.5 years after the fourth vaccination . However, protection occurs also by memory immunity . Herd immunity is sufficient among the general population, but not among orthodox reformed individuals . Immunosurveillance is an efficient way to evaluate the effects of immunisation programmes and identify risk groups for infection.

Ann Trop Paediatr, 2002 Dec, 22(4), 347 - 53
Evaluation of polymerase chain reaction (PCR) for diagnosing Haemophilus influenzae b meningitis; Singhi SC et al.; A polymerase chain reaction (PCR) for detecting Hib in cerebrospinal fluid (CSF) was evaluated and compared with culture and a latex agglutination test (LAT) in a hospital-based prospective surveillance . We studied 107 children aged from 1 month to 12 years with a clinical and CSF profile suggestive of acute bacterial meningitis . CSF culture was performed on blood-chocolate agar by standard technique, LAT by a commercially available kit (Wellcogen) and PCR using total DNA extracted from CSF samples . Of 107 children, 79% had received one or more doses of injectable antibiotics . Hib was detected by culture in 14 cases, by LAT in 23 and by PCR in 37 . All CSF samples that reveal Hib by culture or LAT had a PCR positive for Hib (sensitivity 100%) . PCR also detected 14 additional cases of Hib meningitis which were not detected by culture or LAT . We conclude that PCR is a sensitive and specific diagnostic tool that may be valuable in a population with high pre-hospital antibiotic usage.

Ugeskr Laeger, 2002 Dec 30, 165(1), 34 - 7
{Initial antibiotic therapy of purulent meningitis in adults . An investigation of practice patterns at Danish hospital department in 2000}; Meyer CN; INTRODUCTION: A previous report demonstrated profound variation in the recommended empirical antibiotic therapy for adult purulent meningitis in Denmark . At present, the only existing "official" recommendation (from the Danish Medical Association) is penicillin monotherapy for all adults, irrespective of age, as the prevalence of penicillin-resistant pneumococci is less than 3% . MATERIAL AND METHODS: A questionnaire concerning empirical antibiotic therapy and the treatment of specific microorganisms was sent to the 125 departments of internal medicine, infectious diseases, clinical microbiology, neurosurgery, otorhinolaryngology, and neurology in 2001 . RESULTS: Consensus was not found in the 93% who replied, neither within or between the medical specialties nor within or between the counties . The departments of medicine and clinical microbiology were evenly distributed between two strategies: 1) a third-generation cephalosporin plus a penicillin, or 2) penicillin monotherapy, in supplemented in few departments with an aminoglycoside . For Haemophilus influenzae meningitis, 20% of the departments used ampicillin, whereas the majority preferred ceftriaxone . DISCUSSION: Consensus on empirical antibiotic treatment of purulent meningitis in adults had still not been reached in Denmark in the year 2000, and regimens other than that recommended by the Danish Medical Association were used . Complete and updated information is lacking on the resistance-patterns of bacteria inducing meningitis, and a complete national clinical microbiological database could form the basis for an evidence-based national consensus.

J Biol Chem, 2003 Mar 21, 278(12), 10790 - 8 Epub 2003 Jan 14.
The tetrameric structure of Haemophilus influenza hybrid Prx5 reveals interactions between electron donor and acceptor proteins; Kim SJ et al.; Cellular redox control is often mediated by oxidation and reduction of cysteine residues in the redox-sensitive proteins, where thioredoxin and glutaredoxin (Grx) play as electron donors for the oxidized proteins . Despite the importance of protein-protein interactions between the electron donor and acceptor proteins, there has been no structural information for the interaction of thioredoxin or Grx with natural target proteins . Here, we present the crystal structure of a novel Haemophilus influenza peroxiredoxin (Prx) hybrid Prx5 determined at 2.8-A resolution . The structure reveals that hybrid Prx5 forms a tightly associated tetramer where active sites of Prx and Grx domains of different monomers interact with each other . The Prx-Grx interface comprises specific charge interactions surrounded by weak interactions, providing insight into the target recognition mechanism of Grx . The tetrameric structure also exhibits a flexible active site and alternative Prx-Grx interactions, which appear to facilitate the electron transfer from Grx to Prx domain . Differences of electron donor binding surfaces in Prx proteins revealed by an analysis based on the structural information explain the electron donor specificities of various Prx proteins.

Chest, 2003 Jan, 123(1), 174 - 80
Microbiological testing and outcome of patients with severe community-acquired pneumonia; Rello J et al.; STUDY OBJECTIVES: The study documents the impact of microbiological investigations on therapeutic decisions and outcome in patients with severe community-acquired pneumonia (SCAP) . DESIGN: Retrospective analysis of prospectively collected data . SETTING: ICUs in two teaching Spanish hospitals . PATIENTS: Two hundred four consecutive patients admitted to intensive care with SCAP . INTERVENTIONS: None . Measurements and results: One hundred six patients required intubation, while 98 other patients did not (81 of these patients were managed with noninvasive mechanical ventilation) . The microbiologic diagnosis was established in 57.3% of patients . The most common pathogens were Streptococcus pneumoniae, Legionella pneumophila, and Haemophilus influenzae . Pseudomonas (6.6.% vs 1.0%, p < 0.05) and Legionella (15.1% vs 7.1%, p < 0.05) were more frequently documented in intubated patients . Overall mortality was 23.5% (44.3% in intubated patients), with S pneumoniae (n = 7), Pseudomonas aeruginosa (n = 7), and L pneumophila (n = 5) being the most common lethal pathogens . Bacteriological investigation led to changes in antibiotic prescription in 41.6% of patients, including 11 patients (5%) in whom initial treatment was ineffective against the microbial isolates . The most frequent reason for changes was simplification of therapy in 65 episodes (31.8%) . CONCLUSIONS: We conclude that microbiological testing is fully justified in patients with SCAP, because identifying the causative agent and adjusting treatment both impact on patient outcome . Our findings suggest that intubated patients should be empirically treated for Pseudomonas and Legionella while awaiting bacteriology results.

Eur J Clin Microbiol Infect Dis, 2002 Dec, 21(12), 856 - 63 Epub 2002 Dec 10.
Do HIV-seropositive patients become colonised with drug-resistant microorganisms?
Leegaard TM, Caugant DA, Froholm LO, Hoiby EA, Ronning EJ, Sandven P, Bruun JN.
The aim of the present study was to investigate whether HIV-infected patients, a group that is supposedly at risk for infection with antibiotic-resistant microbes, really does so, and to assess possible risk factors for acquiring these organisms . During the period from January 1998 to July 1999, samples of normal flora were obtained from 107 HIV-infected patients attending an outpatient clinic in Oslo, Norway . The samples were cultured for Streptococcus pneumoniae, Haemophilus influenzae, Escherichia coli, coagulase-negative staphylococci and Candida spp., and the resulting isolates were tested for antimicrobial susceptibility . The patients studied represented all stages of HIV infection, from recently infected to severely immunocompromised . Samples were taken at one, two or three time-points to determine whether antimicrobial resistance in colonising microorganisms increases over time . Antimicrobial resistance was linked primarily to antimicrobial prophylaxis, but it did not increase during the observation period . The level of a patient's immunodeficiency and the consequently intensified medical care was also of some importance . Even though about 50% of the patients were receiving antimicrobial agents at the time of sampling, the level of resistance found in these patients was very similar to that found in other patient groups in Norway; except for Candida albicans isolates, which were less susceptible to fluconazole . Overall, antimicrobial resistance was uncommon in the HIV-seropositive patients studied, a finding that is probably related to the overall low prevalence of antimicrobial resistance in the general population in Norway.

EMBO Rep, 2003 Jan, 4(1), 47 - 52
The Gam protein of bacteriophage Mu is an orthologue of eukaryotic Ku; d'Adda di Fagagna F et al.; Mu bacteriophage inserts its DNA into the genome of host bacteria and is used as a model for DNA transposition events in other systems . The eukaryotic Ku protein has key roles in DNA repair and in certain transposition events . Here we show that the Gam protein of phage Mu is conserved in bacteria, has sequence homology with both subunits of Ku, and has the potential to adopt a similar architecture to the core DNA-binding region of Ku . Through biochemical studies, we demonstrate that Gam and the related protein of Haemophilus influenzae display DNA binding characteristics remarkably similar to those of human Ku . In addition, we show that Gam can interfere with Ty1 retrotransposition in Saccharomyces cerevisiae . These data reveal structural and functional parallels between bacteriophage Gam and eukaryotic Ku and suggest that their functions have been evolutionarily conserved.

Vet Microbiol, 2003 Apr 2, 92(3), 253 - 62
Typing of Haemophilus parasuis strains by PCR-RFLP analysis of the tbpA gene; de la Puente Redondo VA et al.; On the basis of a species-specific PCR assay, a RFLP analysis for typing of Haemophilus parasuis strains was developed and evaluated . Amplification was based on the gene tbpA, encoding a transferrin-binding protein . RFLP analysis of the 1.9-kb tbpA-amplicon using TaqI, AvaI and RsaI endonucleases produced 12 different patterns for the reference strains of the 15 known H . parasuis serovars, and showed a high heterogeneity (33 RFLP groups) for 101 H . parasuis clinical isolates tested . The sensitivity, typeability (100% versus 65% for immunodiffusion), high degree of discrimination (0.93 versus 0.84 for immunodiffusion), simplicity and low cost per test make this PCR-RFLP assay a useful method for typing H . parasuis and, therefore, for studying the epidemiology of outbreaks of Glasser's disease.

Clin Infect Dis, 2003 Jan 15, 36(2), 189 - 94 Epub 2002 Dec 31.
Indications for the immunological evaluation of patients with meningitis; Overturf GD; Although people with bacterial meningitis lack adequate protective antibody against the invading pathogen, most do not have an underlying immunodeficiency . Certain comorbid conditions increase the risk for development of bacterial sepsis and meningitis . In addition, certain congenital complement deficiencies, defects of antibody production, or asplenia may be first recognized by the occurrence of bacterial meningitis, particularly when it occurs in infants or young children . Deficiencies of the terminal components of complement (C5-C9) or properdin have been associated with recurrent or invasive neisserial infections, and asplenia, agammaglobulinemia, and deficiencies of the early components of complement (e.g., C1-C3) are associated with risks of infections caused by Streptococcus pneumoniae, Haemophilus influenzae, and meningococci . The presence of congenital or acquired immunodeficiencies should be considered in persons who present with bacterial meningitis on the basis of the etiology, clinical epidemiology, and presence of other risk factors.

Curr Microbiol, 2003 Feb, 46(2), 103 - 8
Preferential inhibition of protein synthesis by ketolide antibiotics in Haemophilus influenzae cells; Champney WS et al.; The ketolide antibiotics are semi-synthetic derivatives of erythromycin A with enhanced inhibitory activity in a wide variety of microorganisms . They have significantly lower MICs than the macrolide antibiotics for many Gram-positive organisms . Two ketolides, telithromycin and ABT-773, were tested for growth-inhibitory effects in Haemophilus influenzae . Both antibiotics increased the growth rate and reduced the viable cell number with IC(50) values of 1.5 microgram/ml . Protein synthesis was inhibited in cells with a similar IC(50) concentration (1.25 microgram/ml) . Macrolide and ketolide antibiotics have been shown to have a second equivalent target for inhibition in cells, which is blocking the assembly of the 50S ribosomal subunit . Pulse and chase labeling assays were conducted to examine the effect of the ketolides on subunit formation in H . influenzae . Surprisingly, both antibiotics inhibited 50S and 30S subunit assembly to the same extent, with no specific effect of the compounds on 50S assembly . Over a range of antibiotic concentrations, 30S particle synthesis was diminished to the same extent as 50S formation . H . influenzae cells seem to have only one significant target for these antibiotics, and this may help to explain why these drugs are not more effective than the macrolides in preventing the growth of this microorganism.

Clin Microbiol Infect, 2002, 8 Suppl 3, 1 - 8; discussion 33-5
Is antimicrobial resistance also subject to globalization?
Schito GC.
In recent years one of the more alarming aspects of clinical microbiology has been the dramatic increase in the incidence of resistance to antibacterial agents among pathogens causing nosocomial as well as community-acquired infections . There are profound geographic differences in the incidence of resistance among pathogens of the respiratory tract, only some of which can be explained by the local use of antibiotics . A high percentage of Moraxella catarrhalis strains produce beta-lactamase and are thus resistant to many beta-lactam antibiotics . In contrast, beta-lactamase production among strains of Haemophilus influenzae rarely reaches more than 30% around the world . Methicillin-resistance in Staphylococcus aureus is a common and increasing problem in hospitals but its extent varies both locally and nationally . Resistance is usually associated with the local spread of resistant strains . High standards of hygiene in hospitals can prevent the spread of such strains but once established they can be difficult to eradicate . Although Streptococcus pyogenes remains highly susceptible to penicillins, even after many decades of their use, resistance to macrolides has occurred . This resistance can rise and fall . Although the increase of macrolide resistance in S . pyogenes can often be associated with an increase in the use of these drugs, this is not always so . In some cases it has been shown to be caused by the spread of one or more resistant clones . Eradication of these clones can reduce the level of resistance markedly . Resistance to both macrolides and penicillins among strains of Streptococcus pneumoniae is seen world-wide but is highly variable from country to country . Local habits of drug usage may play a part . In Italy, for example, there is preference for the use of parenteral third-generation cephalosporins for some severe infections and there is a corresponding low level of penicillin-resistance among pneumococci.

J Clin Microbiol, 2003 Jan, 41(1), 393 - 6
Identification of Haemophilus influenzae serotypes by standard slide agglutination serotyping and PCR-based capsule typing; LaClaire LL et al.; To resolve discrepancies in slide agglutination serotyping (SAST) results from state health departments and the Centers for Disease Control and Prevention (CDC), we characterized 141 of 751 invasive Haemophilus influenzae isolates that were identified in the United States from January 1998 to December 1999 through an active, laboratory-based, surveillance program coordinated by the CDC . We found discrepancies between the results of SAST performed at state health departments and those of PCR capsule typing performed at the CDC for 56 (40%) of the isolates characterized: 54 isolates that were identified as a particular serotype by SAST were shown to be unencapsulated by PCR, and two isolates that were reported as serotypes b and f were found to be serotypes f and e, respectively, by PCR . The laboratory error most likely to affect the perceived efficacy of the conjugate H . influenzae type b (Hib) vaccine was the misidentification of isolates as serotype b: of 40 isolates identified as serotype b by SAST, 27 (68%) did not contain the correlating capsule type genes . The frequency of errors fell substantially when standardized reagents and routine quality control of SAST were used during a study involving three laboratories . An overall 94% agreement between SAST and PCR results showed that slide agglutination could be a valid and reliable method for serotyping H . influenzae if the test was performed correctly, in accordance with standardized and recommended procedures . An ongoing prospective analysis of all H . influenzae surveillance isolates associated with invasive disease in children less than 5 years old will provide more accurate national figures for the burden of invasive disease caused by Hib and other H . influenzae serotypes.

Clin Invest Med, 2002 Dec, 25(6), 243 - 51
Simultaneous administration of meningococcal C conjugate vaccine and diphtheria-tetanus-acellular pertussis-inactivated poliovirus-Haemophilus influenzae type b conjugate vaccine in children: a randomized double-blind study; Halperin SA et al.; BACKGROUND: Meningococcal C disease can be life-threatening in infants, young children and adolescents . New conjugate vaccines are immunogenic in young infants and induce immunologic memory, so we should consider incorporating them into the routine childhood immunization program . The objective of this study was to measure the safety and immunogenicity of a meningococcal C conjugate vaccine when given with routine childhood vaccines . METHODS: We carried out a randomized, double-blind, controlled clinical trial at children's hospitals in 3 Canadian cities . A convenience sample of 351 healthy 2-month-old infants was enrolled from the community and randomly allocated to receive either meningococcal C conjugate vaccine or the control (hepatitis B) vaccine . All participants received a concurrent injection of the combined diphtheria-tetanus-acellular pertussis-inactivated poliovirus-Haemophilus influenzae type b (DTaP-IPV-Hib) conjugate vaccine in the opposite limb . Participants were immunized at 2, 4, 6 and 15 months of age; adverse events were recorded after each dose . Serum bactericidal and ELISA meningococcal antibody levels in the participants were measured at 6, 7, 15 and 16 months of age; diphtheria, tetanus, H . influenzae type b, poliovirus and pertussis antibodies were measured at 7 months of age . A total of 323 (92%) participants completed all aspects of the study . The proportion of participants who suffered adverse events after each vaccine dose was the primary safety outcome . Geometric mean antibody titres and the proportion of participants with protective antibody levels after immunization were the primary immunologic outcomes . RESULTS: After 2 doses of the meningococcal C conjugate vaccine 99% of participants achieved a protective ( > or = 1:8) bactericidal meningococcal serogroup C antibody level, and after 3 doses this rate increased to 100% . Antibody levels to the concomitant vaccine antigens in the group receiving meningococcal C vaccine were similar to those in the control group except for higher antidiphtheria antibody titres (p < 0.001) . Local injection site reactions (redness and induration) after the meningococcal conjugate vaccine were more frequent than after hepatitis B vaccine but less frequent than after the DTaP-IPV-Hib vaccine . CONCLUSIONS: The meningococcal C conjugate vaccine can be safely and effectively administered at the same visit as the other vaccine antigens routinely given to infants in Canada.

Przegl Lek, 2002, 59(7), 486 - 8
{Bacterial flora of hypertrophied tonsils in children with secretory otitis media}; Wysocka J et al.; The potentially pathogenic bacteria: Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pneumoniae are the major bacterial flora of hypertrophied tonsils . In children suffering from otitis media with inflammatory effusion an unfortunate decreased physiological flora of Streptococcus viridans and Neisseria is observed.

Otolaryngol Head Neck Surg, 2002 Dec, 127(6 Suppl), S17 - 23
Activity of oral beta-lactam antimicrobial agents versus respiratory tract isolates of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis in the era of antibiotic resistance; Doern GV; The management of outpatient respiratory tract infections with oral beta-lactam antimicrobial agents has been complicated by the emergence of beta-lactamase-producing strains of Haemophilus influenzae and Moraxella catarrhalis and isolates of Streptococcus pneumoniae that express varying levels of beta-lactam resistance because of altered penicillin-binding proteins . There is no question that all 3 of these important respiratory tract pathogens have changed conspicuously in the context of decreased activity of beta-lactam antimicrobial agents . The question arises, what does this change mean from a clinical perspective? In this review, the in vitro activity of various oral beta-lactam antimicrobial agents versus contemporary isolates of H influenzae, M catarrhalis, and S pneumoniae will be compared . The approach that is used in defining organisms as being susceptible or resistant to beta-lactam antimicrobial agents will be elucidated . Finally, an effort will be made to assess the ramifications of beta-lactam resistance for therapeutic efficacy in patients with respiratory tract infections caused by these 3 bacteria . We conclude that, notwithstanding diminished beta-lactam activity for many clinical isolates of H influenzae, M catarrhalis, and S pneumoniae, certain agents remain effective in treating outpatient respiratory tract infections.

Otolaryngol Head Neck Surg, 2002 Dec, 127(6 Suppl), S3 - 9
Issues in the management of bacterial sinusitis; Bishai WR; Office visits for acute bacterial rhinosinusitis (ABRS) have increased steadily in the past 2 decades, and ABRS accounted for 0.4% of ambulatory diagnoses in 1995, ranking as the fifth most common diagnosis for which an antibiotic is prescribed . ABRS typically begins as a viral respiratory tract infection, but bacterial growth is demonstrated in 60% of adults with upper respiratory tract infection symptoms of at least 10 days duration . Important factors to consider when selecting an antibiotic regimen for ABRS include: severity of disease, rate of disease progression, earlier antibiotic treatment, regional resistance rates, and, in children, an age <5 years and attendance in day care centers . The most prevalent causative pathogens of sinusitis are Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis . Antimicrobial agent use is under significant scrutiny in this era of emerging bacterial resistance, and antibiotic treatment for sinusitis should include consideration of drug pharmacodynamics . Several agents, once considered first-line therapies, can no longer be considered dependable in this role . Rather, newer antibiotics may be called on to provide efficacy and forestall additional antimicrobial resistance among respiratory tract pathogens.

J Infect Dis, 2003 Jan 1, 187(1), 109 - 16 Epub 2002 Dec 13.
Prevention of Haemophilus influenzae type b (Hib) meningitis and emergence of serotype replacement with type a strains after introduction of Hib immunization in Brazil; Ribeiro GS et al.; Surveillance for Haemophilus influenzae meningitis cases was performed in Salvador, Brazil, before and after introduction of H . influenzae type b (Hib) immunization . The incidence of Hib meningitis decreased 69% during the 1-year period after initiation of Hib immunization (from 2.62 to 0.81 cases/100,000 person-years; P<.001) . In contrast, the incidence for H . influenzae type a meningitis increased 8-fold (from 0.02 to 0.16 cases/100,000 person-years; P=.008) . Pulsed-field gel electrophoretic analysis demonstrated that H . influenzae type a isolates belonged to 2 clonally related groups, both of which were found before Hib immunization commenced . Therefore, Hib immunization contributed to an increased risk for H . influenzae type a meningitis through selection of circulating H . influenzae type a clones . The risk attributable to serotype replacement is small in comparison to the large reduction in Hib meningitis due to immunization . However, these findings highlight the need to maintain surveillance as the use of conjugate vaccines expands worldwide.

Res Vet Sci, 2003 Feb, 74(1), 23 - 30
Influences of naturally occurring and experimentally induced porcine pneumonia on blood parameters; Muller G et al.; It had been the objective of the studies described to establish local and systemic changes by naturally occurring pneumonia or pneumonia experimentally induced by Pasteurella multocida and Haemophilus parasuis in swine . Acute and chronic pneumonia was found to alter the cytokine level of lung lavage fluid and affect the composition and function of blood cells, especially with regard to phagocytosis, radical formation and cell surface receptors . Interleukin-6 levels in blood plasma rose 24h after experimental intrabronchial infection . The influences of the changes on growth and meat quality are discussed.

S Afr Med J, 2002 Nov, 92(11), 907 - 11
Aetiology and outcome of severe community-acquired pneumonia in children admitted to a paediatric intensive care unit; Delport SD et al.; OBJECTIVE: To determine the aetiological agents and outcome of severe community-acquired pneumonia (SCAP) in children admitted to the paediatric intensive care unit (PICU) at Kalafong Hospital, Pretoria . PATIENTS AND METHODS: An audit was done after a protocol was implemented to identify the aetiological agents in children with life-threatening SCAP admitted to the PICU from the emergency room . The following investigations were done as per protocol: blood culture, culture of the tracheal aspirate, immunofluorescence and culture of the nasopharyngeal aspirate, microscopy and culture of the gastric juice for Mycobacterium tuberculosis, and determination of HIV status . The following data, documented prospectively, were obtained from patient records: date of admission, age, gender, weight, duration of ventilation, duration of stay in the PICU, survival or death, and severity of illness as determined by means of the score for acute neonatal physiology (SNAP) or paediatric risk of mortality (PRISM) score depending on the child's age . RESULTS: Twenty-three children were admitted over a 1-year period (1 November 1994-30 October 1995) . Their median age was 10 weeks (range 2 weeks-5 years) and the sex distribution was equal . Two children were HIV-infected . Twenty children received mechanical ventilation for a median period of 6.5 days (range 2-16 days) . Aetiological agents were identified in 15/23 children (65%) . Respiratory syncytial virus (RSV) was the most common pathogen, identified in 7/23 children, Klebsiella pneumoniae was the most common bacterial pathogen, identified in 5 children (2 blood cultures and 3 tracheal aspirates) . Tuberculosis was not diagnosed . The mean PRISM score was similar in survivors and children who died . The case fatality rate was 30% . The 7 children who died had a median arterial oxygen tension/fraction of inspired oxygen (PaO2/FiO2) ratio of 94 (range 32-111) and the 16 survivors had a median ratio of 146 (range 51-252) (P = 0.01) on admission . Both HIV-infected children died and postmortem examination showed a pneumonia due to Pneumocystis carinii and cytomegalovirus . CONCLUSIONS: SCAP occurs in very young children . One or more pathogens were isolated in 65% of cases . Viral pathogens predominated, with RSV being the most common . The yield of positive blood cultures was low at 17% . Streptococcus pneumoniae and Haemophilus influenzae were not found . The case fatality rate was 30% and death was more likely with a low PaO2/FiO2 ratio on admission.

Blood, 2003 Apr 15, 101(8), 3319 - 24 Epub 2002 Dec 27.
Factors affecting antibody levels after allogeneic hematopoietic cell transplantation; Storek J et al.; To obtain insight into the mechanism(s) of posttransplantation humoral immunodeficiency, we evaluated factors affecting serum antibody levels against polio, tetanus, Haemophilus influenzae, and Streptococcus pneumoniae in 87 patients . Patients with hematologic malignancies were randomized to receive marrow versus blood stem cells, which contain approximately 10 times more lymphocytes than marrow . Blood stem cell recipients did not have higher antibody levels than marrow recipients . Recipient pretransplantation antibody levels were correlated with the posttransplantation levels, especially in the first 6 months after transplantation when the correlation coefficients typically exceeded 0.6 . Donor pretransplantation antibody levels had less of a correlation with posttransplantation levels in the recipient . Patient or donor age, total body irradiation, and graft-versus-host disease or its treatment appeared to have no effect . In conclusion, antibody levels in the first year after transplantation are affected primarily by pretransplantation antibody levels in the recipient and, to a lesser degree, in the donor . These findings suggest that immunization of the recipient and the donor before transplantation may be more effective in improving antibody immunity after transplantation than manipulating graft-versus-host disease, changing conditioning, or increasing the number of lymphocytes in the graft.

J Infect, 2003 Jan, 46(1), 46 - 8
Changing socioeconomic risk factors for invasive H . influenzae disease after the introduction of conjugate vaccine; Olowokure B et al.; OBJECTIVES: To determine socioeconomic risk factors for invasive Haemophilus influenzae (HI) disease before and after the introduction of HI type b (Hib) conjugate vaccine in the West Midlands, UK . METHODS: Study population consisted of children aged <5 years with laboratory confirmed invasive disease . Linear associations between disease rates and socioeconomic variables were examined 2-years before and 2-years after the introduction of conjugate vaccine . RESULTS: Pre-vaccine there were no trends in disease incidence . Post-vaccine there was an absolute reduction in disease incidence and significant disease differentials between affluent and deprived populations were identified for non-owner-occupancy (P=0.032) and car access (P=0.049) . CONCLUSIONS: Our results suggest that changes in socioeconomic risk factor for invasive HI occurred after the introduction of conjugate vaccine . This may have implications for future immunisation strategies.

Clin Ther, 2002 Nov, 24(11), 1854 - 70
A comparison of cefditoren pivoxil and amoxicillin/ clavulanate in the treatment of community-acquired pneumonia: a multicenter, prospective, randomized, investigator-blinded, parallel-group study; Fogarty CM et al.; BACKGROUND: Cefditoren pivoxil is a broad-spectrum cephalosporin that is approved for the treatment of pharyngitis, acute exacerbations of chronic bronchitis, and skin and skin-structure infections . OBJECTIVE: This study was conducted to examine the efficacy and tolerability of cefditoren in the treatment of community-acquired pneumonia (CAP) . Amoxicillin/clavulanate was chosen as a comparator because of its established efficacy and general acceptance as a standard of care in CAP . METHODS: This multicenter, prospective, randomized, investigator-blinded, parallel-group trial compared oral cefditoren 200 and 400 mg BID with oral amoxicillin/clavulanate 875/125 mg BID for 14 days in adult outpatients with CAP . RESULTS: Eight hundred two patients (404 men, 398 women; mean age, 50 years; age range, 12-93 years) with CAP were enrolled . Comparable clinical cure rates were observed among evaluable patients in all treatment groups at both the posttreatment and follow-up visits: 88.0% (125/142) for cefditoren 200 mg, 89.9% (143/159) for cefditoren 400 mg, and 90.3% (130/144) for amoxicillin/clavulanate at the posttreatment visit, and 86.5% (128/148), 86.8% (138/159), and 87.8% (129/147) for the respective groups at the follow-up visit . Of 82 Streptococcus pneumoniae strains isolated before treatment, 22 (26.8%) had reduced susceptibility to penicillin, 12 (14.6%) of them penicillin resistant . Overall eradication rates at the posttreatment visit for pathogens isolated from microbiologically evaluable patients were 84.0%, 88.6%, and 82.6% for cefditoren 200 mg, cefditoren 400 mg, and amoxicillin/clavulanate, respectively . In the respective treatment groups, 80.6%, 88.6%, and 88.0% of Haemophilus influenzae strains and 95.0%, 96.2%, and 89.5% of S pneumoniae strains were eradicated . The rates of resolution of or improvement in clinical signs and symptoms were comparable between treatment groups . The treatment regimens were well tolerated, with 4.9%, 3.0%, and 5.2% of patients in the respective treatment groups requiring discontinuation of study drug due to an adverse event . CONCLUSIONS: In this study in adult outpatients with CAP, both doses of cefditoren demonstrated equivalence to amoxicillin/clavulanate based on rates of clinical and microbiologic cure . All 3 regimens were effective in resolving or improving the clinical signs and symptoms of CAP . Both cefditoren and amoxicillin/ clavulanate were well tolerated.

Clin Ther, 2002 Nov, 24(11), 1840 - 53
Cefditoren pivoxil versus cefpodoxime proxetil for community-acquired pneumonia: results of a multicenter, prospective, randomized, double-blind study; van Zyl L et al.; BACKGROUND: According to recently issued treatment guidelines, appropriate empiric choices for ambulatory patients with community-acquired pneumonia (CAP) are a macrolide, doxycycline (for patients aged > or = 8 years), or an oral beta-lactam agent with good activity against pneumococci . OBJECTIVE: This study was designed to compare cefditoren pivoxil, a new beta-lactam, with cefpodoxime proxetil, a beta-lactam with an established role in the treatment of CAP . METHODS: This was a multicenter, prospective, randomized, double-blind study conducted in the United States and South Africa . Ambulatory patients with a diagnosis of CAP were randomized to 14 days of treatment with cefditoren 200 or 400 mg BID or cefpodoxime 200 mg BID . Assessments of clinical cure and pathogen eradication were conducted at 2 visits during treatment, 1 posttreatment visit (s48 hours after completion of treatment), and 1 follow-up visit (7-14 days after completion of treatment) . The development of resistant pathogens was assessed at the follow-up visit but not thereafter . The relative cost of treatment was not assessed . RESULTS: The study enrolled 851 patients . Comparable clinical cure rates were observed among evaluable patients in the 3 treatment groups at both the posttreatment and followup visits: at the posttreatment visit, cure rates were 90.5% (162/179) for cefditoren 200 mg, 89.7% (148/165) for cefditoren 400 mg, and 92.2% (153/166) for cefpodoxime 200 mg; at the follow-up visit, they were a respective 88.4% (160/181), 87.2% (143/164), and 90.4% (151/167) . Of the 171 strains of Streptococcus pneumoniae isolated before treatment, 22 (12.9%) had reduced susceptibility to penicillin, 5 (2.9%) of them penicillin resistant (minimum inhibitory concentration > or = 2 microg/mL) . At the posttreatment visit, the overall eradication rates of pathogens isolated from microbiologically evaluable patients were 88.7% (134/151), 89.9% (134/149), and 95.7% (134/140) in the respective treatment groups (P = 0.031, cefditoren 200 mg vs cefpodoxime) . Eradication rates of S pneumoniae were 93.8% (45/48), 95.7% (45/47), and 95.6% (43/ 45) in the respective treatment groups; those of Haemophilus influenzae were 90.2% (46/51), 97.7% (43/44), and 97.4% (37/38) . The rates of resolution and/or improvement in clinical signs and symptoms were comparable between groups . The study drugs were well tolerated, with 1.7%, 2.5%, and 1.4% of patients in the respective groups discontinuing study drug prematurely due to a treatment-related adverse event, the majority of these associated with the digestive system . CONCLUSION: The results of this study suggest that cefditoren may have a role in the treatment of CAP in ambulatory patients.

Acta Crystallogr D Biol Crystallogr, 2003 Jan, 59(Pt 1), 183 - 4 Epub 2002 Dec 19.
Crystallization and preliminary X-ray crystallographic analysis of tRNA(m1G37)methyltransferase from Haemophilus influenzae; Kim HW et al.; The enzyme tRNA(m(1)G37)methyltransferase (TrmD) catalyzes the transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) specifically to guanosine at position 37 within a subset of tRNA species in bacteria . The modified guanosine is next to the anticodon and is important for the maintenance of the correct reading frame during translation . TrmD from Haemophilus influenzae with both N- and C-terminal tags was overexpressed in Escherichia coli and crystallized at 297 K using sodium acetate as a precipitant . Native X-ray diffraction data were collected to 1.85 A resolution . The crystals are rhombohedral, belonging to the space group R32, with unit-cell parameters a = b = 98.05, c = 176.79 A, alpha = beta = 90, gamma = 120 degrees . The presence of one monomer of recombinant TrmD in the crystallographic asymmetric unit gives a V(M) of 3.07 A(3) Da(-1) and a solvent content of 59.9%.

Acta Crystallogr D Biol Crystallogr, 2003 Jan, 59(Pt 1), 180 - 2 Epub 2002 Dec 19.
Crystallization and preliminary X-ray crystallographic studies of 3-deoxy-manno-octulosonate cytidylyltransferase from Haemophilus influenzae; Ku MJ et al.; The enzyme 3-deoxy-manno-octulosonate cytidylyltransferase (CMP-KDO synthetase; CKS) catalyzes the activation of 3-deoxy-manno-octulosonate (KDO) by forming CMP-KDO . It is essential for the biosynthesis of lipopolysaccharides in Gram-negative bacteria and is a potential target for the discovery of antibacterial agents . L-CKS from Haemophilus influenzae was overexpressed with a C-terminal hexahistidine tag in Escherichia coli and crystallized in the presence of the substrate KDO at 297 K using PEG 4000 as a precipitant and ethylene glycol as an additive . The diffraction limit and spot shape of the native crystal could be improved significantly by dehydration/annealing . X-ray diffraction data were collected to 2.5 A resolution from a native crystal . The crystals are orthorhombic, belonging to the space group P2(1)2(1)2(1), with unit-cell parameters a = 48.6, b = 83.1, c = 117.3 A . The presence of two monomers of recombinant L-CKS in the crystallographic asymmetric unit gives a reasonable V(M) of 2.05 A(3) Da(-1), with a solvent content of 40.0%.

Antimicrob Agents Chemother, 2003 Jan, 47(1), 405 - 7
Activities of HMR 3787 and RU 64399 compared with those of four other agents against Haemophilus influenzae and Haemophilus parainfluenzae; Bozdogan B et al.; Activities of HMR 3787, a new 2-fluoroketolide, and its (des)-fluor derivative, RU 64399, were tested against 111 Haemophilus influenzae and 26 H . parainfluenzae strains and compared with those of telithromycin, erythromycin, azithromycin, and clarithromycin . HMR 3787 and RU 64399 MICs were comparable with those of azithromycin but were less affected by incubation in CO(2) . Time-kill studies of 12 strains showed that HMR 3787, RU 64399, and telithromycin were bactericidal against all strains after 24 h at two times the MIC.

Can Vet J, 2002 Nov, 43(11), 863 - 8
Coinfection with bovine viral diarrhea virus and Mycoplasma bovis in feedlot cattle with chronic pneumonia; Shahriar FM et al.; Chronic, antibiotic-resistant pneumonia, sometimes with concurrent polyarthritis, occurs in feedlot cattle in western Canada . The prevalence of Mycoplasma bovis, bovine viral diarrhea virus, and Haemophilus somnus was determined by using immunohistochemical staining of lung and heart tissue from 2 groups of animals with this history . Mycoplasma bovis antigen was present in 44/48 cases submitted between 1995 and 1998 (retrospective group) and 15/16 of cases from 1999 (prospective group), and was associated with pulmonary necrosis . Bovine viral diarrhea virus antigen was present in association with microscopic vascular lesions in 31/48 retrospective and 9/16 of prospective cases . Types Ib and II bovine viral diarrhea virus were isolated from 4/16 prospective cases . Haemophilus somnus antigen was present in heart, lung, or both of 15/48 retrospective and 8/16 prospective cases . The results suggest that there may be synergism between bovine viral diarrhea virus and M . bovis in this pneumonia with arthritis syndrome.

Arch Dis Child Fetal Neonatal Ed, 2003 Jan, 88(1), F58 - 61
Effect of a fourth Haemophilus influenzae type b immunisation in preterm infants who received dexamethasone for chronic lung disease; Clarke P et al.; Aim: To assess whether a fourth Hib polysaccharide-tetanus protein conjugate vaccine (PRP-T) would improve antibody response in preterm infants previously treated with dexamethasone for chronic lung disease . METHODS: In a pilot study 12 infants born at less than 30 weeks gestation who had received corticosteroids were given a supplementary Hib dose six weeks after completion of the primary immunisation course . Serum samples obtained before and at eight weeks following the fourth Hib dose were analysed for total level and avidity of anti-PRP antibody . RESULTS: There was no significant increase in the geometric mean titre (GMT) of anti-PRP antibody resulting from the fourth Hib immunisation (GMT: pre 2.35 micro g/ml, post 2.24 micro g/ml, p = 0.79) . A subgroup of six infants had subprotective antibody levels (<1.0 micro g/ml) after the primary immunisation course, which remained subprotective following the extra Hib immunisation . Despite the poor response in total antibody level, the study group showed a significant rise in PRP specific IgG avidity following the fourth immunisation (GMAI: pre 0.076, post 0.138, p = 0.043) . CONCLUSION: An additional Hib immunisation given to recently steroid treated preterm infants six weeks after completion of the primary schedule did not augment primary immunogenicity . However, increasing avidity may imply successful priming and long term immunity to Hib.

Infect Immun, 2003 Jan, 71(1), 163 - 72
Two-component systems in Haemophilus influenzae: a regulatory role for ArcA in serum resistance; De Souza-Hart JA et al.; Knockout mutations were constructed in the arcA gene of a virulent type b strain of Haemophilus influenzae, and the behavior of the resulting mutants was investigated in a number of conditions that mimicked distinct steps in the natural infection pathway . In arcA mutants, synthesis of capsule and lipooligosaccharide (LOS) and growth in synthetic media were unaltered compared to synthesis of capsule and LOS and growth in synthetic media in the wild-type H . influenzae type b parent strain . However, the virulence of the arcA mutants for BALB/c mice was significantly reduced . Upon exposure to human blood or serum, the arcA mutants showed markedly reduced survival compared with the survival of its wild-type parent . Serum resistance could be fully restored by complementation in cis with the H . influenzae arcA gene but not by complementation in cis with the homologous gene from Escherichia coli . The proteomes of wild-type and mutant bacteria were markedly different, especially under anaerobic conditions, underscoring the global regulatory role of ArcAB in H . influenzae . Evaluation of antibody titers and classical complement activities in various serum samples pointed to complement-mediated bactericidal activity as the factor that distinguishes between the arcA mutant and wild-type phenotypes . Comparative analysis of the membrane fractions of the arcA mutants and the wild-type strain revealed several ArcA-regulated proteins, some of which may be implicated in the serum hypersensitivity phenotype.

Braz J Infect Dis, 2002 Oct, 6(5), 206 - 18
A multicenter comparative study of cefepime versus broad-spectrum antibacterial therapy in moderate and severe bacterial infections; Badaro R et al.; The safety and efficacy of cefepime empiric monotherapy compared with standard broad-spectrum combination therapy for hospitalized adult patients with moderate to severe community-acquired bacterial infections were evaluated . In an open-label, multicenter study, 317 patients with an Acute Physiology and Chronic Health Evaluation (APACHE II) score ranging from >5 to =19 were enrolled with documented pneumonia (n=196), urinary tract infection (n=65), intra-abdominal infection (n=38), or sepsis (n=18) . Patients were randomly assigned 1:1 to receive cefepime 1 to 2 g IV twice daily or three times a day or IV ampicillin, cephalothin, or ceftriaxone +/-aminoglycoside therapy for 3 to 21 days . For both treatment groups, metronidazole, vancomycin, or macrolide therapy was added as deemed necessary . The primary efficacy variable was clinical response at the end of therapy . Two hundred ninety-six (93%) patients met evaluation criteria and were included in the efficacy analysis . Diagnoses included the following: 180 pneumonias (90 cefepime, 90 comparator), 62 urinary tract infections (29 cefepime, 33 comparator), 37 intra-abdominal infections (19 cefepime, 18 comparator), and 17 sepses (8 cefepime, 9 comparator) . At the end of therapy, overall clinical success rates were 131/146 (90%) for patients treated with cefepime vs 125/150 (83%) for those treated with comparator (95% confidence interval {CI}: -2.6% to 16.3%) . The clinical success rate for patients with community-acquired pneumonia, the most frequent infection, was 86% for both treatment groups . Among the patients clinically evaluated, 162 pathogens were isolated and identified before therapy . The most commonly isolated pathogens were Escherichia coli (n=49), Streptococcus pneumoniae (n=29), Haemophilus influenzae (n=14), and Staphylococcus aureus (n=11) . Bacteriologic eradication/presumed eradication was 97% for cefepime vs 94% for comparator-treated patients . Drug-related adverse events were reported in 16% of cefepime patients and 19% of comparator patients . In conclusion, cefepime had higher cure rates compared with broad-spectrum combination therapy as an initial empiric treatment for hospitalized patients with moderate to severe community-acquired infections, including urinary tract infections, intra-abdominal infections, and sepsis.

J Antimicrob Chemother, 2003 Jan, 51(1), 147 - 51
Activities of 13 quinolones by three susceptibility testing methods against a collection of Haemophilus influenzae isolates with different levels of susceptibility to ciprofloxacin: evidence for cross-resistance; Perez-Vazquez M et al.; The activities of nalidixic acid, ciprofloxacin, norfloxacin, ofloxacin, pefloxacin, flerofloxacin, sparfloxacin, grepafloxacin, gatifloxacin, moxifloxacin, trovafloxacin, levofloxacin and clinafloxacin against a panel of Haemophilus influenzae strains were assessed by three susceptibility testing methods: Etest, agar dilution and the reference broth microdilution method using Haemophilus test medium (HTM) in all cases . The panel included 62 clinical and two reference H . influenzae strains; 32 had decreased susceptibility to ciprofloxacin (MIC > or = 0.12 mg/L) and 30 were susceptible to this antibiotic (MIC < or = 0.06 mg/L) . Both Etest and HTM agar dilution results (r = 0.96; 86.61% and 82.1% of MICs within + one log(2), respectively) correlated well with the reference microdilution method . The MIC(90) of ciprofloxacin was 4.0 mg/L (range 0.007-32.0 mg/L) . Trovafloxacin activity was similar to that of ciprofloxacin but sparfloxacin, grepafloxacin, ofloxacin, pefloxacin and flerofloxacin activities were higher (with MIC values one log(2) dilution lower than ciprofloxacin) . The least active were norfloxacin (MIC(90) 16 mg/L) and nalidixic acid (MIC(90) 128 mg/L) . Levofloxacin and moxifloxacin were more active than ciprofloxacin (MIC(90) 2 mg/L); clinafloxacin and gatifloxacin were the most active with an MIC(90) of 0.25 mg/L . Cross-susceptibility among all quinolones was observed (r > 0.9) . Resistance to ciprofloxacin was associated with a similar magnitude of activity loss to other new and old quinolones . Ciprofloxacin MIC determination should be sufficient to detect the decreased susceptibility to the whole group of quinolones.

J Antimicrob Chemother, 2003 Jan, 51(1), 39 - 43
Effect of extended-spectrum beta-lactamases on the susceptibility of Haemophilus influenzae to cephalosporins; Tristram SG; The extended-spectrum beta-lactamases (ESBLs) TEM-3, TEM-4 and TEM-5 were cloned into Haemophilus influenzae . These recombinant strains exhibited cefotaxime MICs of 0.5, 0.25 and 0.12 mg/L for TEM-3, -4 and -5, respectively, and the MIC of cefaclor was 4.0 mg/L . These MICs are higher than those of beta-lactamase-negative strains, or those producing simple wild-type TEM-1 beta-lactamase, but not high enough to be categorized as resistant according to the breakpoints of the NCCLS . The clones were also categorized as susceptible using NCCLS disc diffusion methodology and interpretive criteria . This study shows that current NCCLS susceptibility testing methods may have difficulty in detecting ESBLs if they were to occur in H . influenzae.

Vet Microbiol, 2003 Mar 20, 92(1-2), 121 - 34
Cloning and characterisation of type 4 fimbrial genes from Actinobacillus pleuropneumoniae; Stevenson A et al.; Actinobacillus pleuropneumoniae is the cause of porcine pleuropneumoniae . Little is known about the mechanisms by which A . pleuropneumoniae colonises the respiratory tract . Fimbriae are common mediators of bacterial adherence to mucosal epithelia and have been observed on the surface of A . pleuropneumoniae cells . Here we report the identification and characterisation of the type 4 fimbrial structural gene (apfA) from A . pleuropneumoniae . In addition a number of open reading frames were identified in A . pleuropneumoniae that have significant homology to type 4 fimbrial biogenesis genes from other species, including a putative leader specific peptidase (apfD) . A . pleuropneumoniae apfA codes for a predicted polypeptide of approximately 16kDa, removal of the leader sequence at the predicted cleavage site would yield a 14.5kDa polypeptide . The first 30 residues of the mature polypeptide are well conserved with other members of the group A type 4 fimbriae family . The signal sequence of ApfA is 13 amino acids in length and, unusually, the residue that precedes the cleavage site is alanine rather than glycine which is found in most other type 4 fimbriae . The C-terminus of ApfA possesses cysteine residues that are conserved in type 4 fimbriae of many species . In other type 4 fimbriae the distal C-terminal cysteines form a disulphide bond that produces a loop, which is important for the function of fimbriae and also comprises a major antigenic determinant . A motif within the predicted loop in ApfA was found to be highly conserved in type 4 fimbriae of other HAP organisms (Haemophilus, Actinobacillus, Pasteurella) . The A . pleuropneumoniae type 4 fimbrial biogenesis genes showed the strongest homology to putative type 4 fimbrial genes of Haemophilus ducreyi . A . pleuropneumoniae apfA gene was shown to be present and highly conserved in different serotypes of A . pleuropneumoniae . Recombinant ApfA was produced and used to raise anti-ApfA antisera.

Proteins, 2003 Feb 1, 50(2), 249 - 60
The HI0073/HI0074 protein pair from Haemophilus influenzae is a member of a new nucleotidyltransferase family: structure, sequence analyses, and solution studies; Lehmann C et al.; The crystal structure of HI0074 from Haemophilus influenzae, a protein of unknown function, has been determined at a resolution of 2.4 A . The molecules form an up-down, four-helix bundle, and associate into homodimers . The fold is most closely related to the substrate-binding domain of KNTase, yet the amino acid sequences of the two proteins exhibit no significant homology . Sequence analyses of completely and incompletely sequenced genomes reveal that the two adjacent genes, HI0074 and HI0073, and their close relatives comprise a new family of nucleotidyltransferases, with 15 members at the time of writing . The analyses also indicate that this is one of eight families of a large nucleotidyltransferase superfamily, whose members were identified based on the proximity of the nucleotide- and substrate-binding domains on the respective genomes . Both HI0073 and HI0074 were annotated "hypothetical" in the original genome sequencing publication . HI0073 was cloned, expressed, and purified, and was shown to form a complex with HI0074 by polyacrylamide gel electrophoresis under nondenaturing conditions, analytic size exclusion chromatography, and dynamic light scattering . Double- and single-stranded DNA binding assays showed no evidence of DNA binding to HI0074 or to HI0073/HI0074 complex despite the suggestive shape of the putative binding cleft formed by the HI0074 dimer .

J Vet Med B Infect Dis Vet Public Health, 2002 Dec, 49(10), 464 - 8
The effect of season and vaccination for Glässer's disease and post-weaning Colibacillosis in an outdoor pig unit endemically infected with virulent strain of Haemophilus Parasuis serotype 5 and pathogenic Escherichia coli; Karg G et al.; The objective of this field trial was to determine if vaccination against Haemophilus parasuis serovar 5 (HPS 5) and pathogenic serotypes of Escherichia coli would improve nursery pig performance in an outdoor unit in different seasons . The unit was concurrently infected with HPS 5 and with different serotypes of E . coli . All piglets were born to HPS 5 vaccinated sows . The trial was carried out in four (two summer and two winter) groups . Group 1 (E . coli and HPS vaccinated, summer season) (n = 362): Piglets were vaccinated pre-weaning with inactivated E . coli-VT2e-toxin and post-weaning against HPS 5 . Group 2 (non-vaccinated, summer season) (n = 349): Piglets were not vaccinated . Group 3 (E . coli and HPS vaccinated, winter season) (n = 358): The animals were analogously treated as Group 1 . Group 4 (non-vaccinated, winter season) (n = 353): Piglets were not vaccinated . The following parameters were evaluated: A: average daily nursery weight gain (ADG), B: nursery mortality, C: feed efficiency (FE) . No significant weight differences were detected within the vaccinated and non-vaccinated summer or winter raised groups of weaners . Summer raised weaners were significantly (P<0.05) heavier from day 35 on than winter raised animals . ADG and FE of summer raised pigs were significantly better (weeks 1-3 P<0.05; fourth week post-weaning P<0.01) during the nursery period than that of the winter raised groups . Winter raised vaccinated weaners showed during the last week of nursing significantly (P<0.05) better daily gain and feed efficiency compared with the non-vaccinated winter raised animals . Non-significant ADG and FE differences were detectable between the summer raised vaccinated or non-vaccinated groups of pig . Winter raised non-vaccinated animals suffered significantly (P<0.05) higher nursery mortality (10.63%) compared to the winter raised vaccinated animals . Implication: In cases of concurrent infections with HPS 5 and with different serotypes of E . coli, especially during winter season, vaccination against both diseases is suggested.

Pediatr Allergy Immunol, 2002 Dec, 13(6), 443 - 7
Evaluation of the relevance of humoral immunodeficiencies in a pediatric population affected by recurrent infections; Finocchi A et al.; Recurrent infections are a common cause of morbidity in childhood . Several reports have associated this condition to low levels of IgA and IgG subclasses and/or lack of specific antipolysaccharide antibody response, although the relevance of these defects in terms of prognosis and therapeutic approach is still unclear . The aim of our study was to determine the frequency and the clinical relevance of humoral immunodeficiency (HID) other than hypogammaglobulinemia in children affected by recurrent infections . We recruited 67 pediatric patients affected by recurrent infections . Serum IgG, IgA, IgM, IgG2, IgG3, and specific anti-Haemophilus influenzae (anti-Hib) antibodies were determined . Thirty-seven out of 67 patients showed antibody defects (55%) . IgA deficiency was observed in 21 out of 67 patients (31%), followed by IgG2 (18%), IgG3 (15%) and IgM (6%) defects . Anti-Hib deficiency was present in three out of 44 patients (7%) . A tendency for a higher occurrence of pneumonia and otitis, although not statistically significant (p > 0.05), was observed in HID patients compared to children with normal humoral function . No statistical difference as to the frequency of mild infections (URI) was found between HID and non-HID patients . We therefore suggest that the therapeutic program is based on the clinical status of the patients . Long-term follow-up with repeated determinations of antibody levels is crucial, however, to detect those defects that might evolve into more complex immunodeficiencies.

Acta Otolaryngol, 2002 Oct, 122(7), 745 - 51
Bacterial adherence to pharyngeal cells: in vitro studies with alpha-haemolytic streptococci and Haemophilus influenzae; Tano K et al.; We examined the adherence to pharyngeal cells of alpha-haemolytic Streptococci (AHS) and Haemophilus influenzae, representing normal flora and otitis media (OM) pathogens, respectively . The bacteria were incubated with epithelial cells brushed from the tonsils, adenoid or tubal orifice of children and adults . Adherence varied among the clinical isolates of AHS and H . influenzae . AHS adhered better to epithelial cells from a child compared with those sampled from an adult . The bacteria adhered better to cells from the tubal orifice compared with those sampled from the adenoid . The selective attachment of AHS to certain cells but not to others could not be correlated to apoptotic/necrotic cells versus viable cells . Incubation of epithelial cells with an isolate of AHS with good inhibitory activity against OM pathogens showed almost no adherence of bacteria to the epithelial cells after 12 and 24 h of incubation . If, however, an isolate of AHS with weak inhibitory activity was incubated with the cells, the bacteria that were attached to the epithelial cells from the beginning showed overgrowth in the broth and increasing attachment to the cells after 12 and 24 h . Thus the inhibitory activity of AHS could also affect the adherence of potential pathogens to the mucosal surfaces . The adherence pattern may at least partially explain the difference in susceptibility to OM between children and adults.

Leuk Lymphoma, 2002 Oct, 43(10), 1967 - 9
Haemophilus influenzae type b (Hib) antibody concentrations and vaccination responses in patients with chronic lymphocytic leukaemia: predicting factors for response; Sinsalo M et al.; We have recently demonstrated a moderate vaccination response rate of 43% against Haemophilus influenzae type b (Hib) conjugate vaccine among adult and elderly patients with chronic lymphocytic leukaemia (CLL) . We now investigated demographic and immunological factors predicting the favourable response and protective antibody concentrations for Hib conjugate vaccine in CLL . Lower age was associated with protective pre- and post-vaccination antibody concentrations . High IgG1 and IgA concentrations were also associated with the protective efficacy . High IgM, in turn, was the best predictor of a significant vaccination response . Again, lower age seemed to be involved in this outcome . Judging from these findings, it would seem beneficial to vaccinate all CLL patients with conjugate vaccines at the presentation of the disease . Investigations of a new pneumococcal conjugate vaccine in CLL are warranted.

Microbiology, 2002 Dec, 148(Pt 12), 3993 - 4001
Differences in iron acquisition from human haemoglobin among strains of Actinobacillus actinomycetemcomitans; Hayashida H et al.; To get a better insight into the physiology of the high-toxic JP2 clone of Actinobacillus actinomycetemcomitans serotype b, which is strongly associated with juvenile periodontitis in adolescents of African descent, the modes of iron acquisition in this clone were examined and compared to those of other strains of the species . None of the strains examined could utilize human transferrin as a source of iron . This was in accordance with the presence of a non-functional tbpA gene, which normally encodes the A subunit of the transferrin-binding-protein complex . Southern blot analysis indicated that functional duplications of tbpA were not present in the genome . Thus, A . actinomycetemcomitans seems to be in a process of evolution, in which iron acquisition from host transferrin is not essential as in many other members of the pasteurellaceae . All strains could utilize haem as a source of iron . All 11 A . actinomycetemcomitans strains examined harboured a single genomic sequence with homology to the hgpA gene encoding haemoglobin-binding protein A in Haemophilus influenzae . However, in all three strains belonging to the JP2 clone and in one serotype e strain hgpA was a pseudogene . Seven other strains possessed a functional hgpA gene which, according to insertion mutagenesis experiments, was responsible for the ability of these strains to utilize haemoglobin as a source of iron . Thus, the presence of an hgpA pseudogene and the inability to use human haemoglobin as an iron source discriminate the high-toxic JP2 clone from low-toxic serotype b strains and most other strains of A . actinomycetemcomitans.

Lancet, 2002 Nov 30, 360(9347), 1737 - 41
Mercury concentrations and metabolism in infants receiving vaccines containing thiomersal: a descriptive study; Pichichero ME et al.; BACKGROUND: Thiomersal is a preservative containing small amounts of ethylmercury that is used in routine vaccines for infants and children . The effect of vaccines containing thiomersal on concentrations of mercury in infants' blood has not been extensively assessed, and the metabolism of ethylmercury in infants is unknown . We aimed to measure concentrations of mercury in blood, urine, and stools of infants who received such vaccines . METHODS: 40 full-term infants aged 6 months and younger were given vaccines that contained thiomersal (diptheria-tetanus-acellular pertussis vaccine, hepatitis B vaccine, and in some children Haemophilus influenzae type b vaccine) . 21 control infants received thiomersal-free vaccines . We obtained samples of blood, urine, and stools 3-28 days after vaccination . Total mercury (organic and inorganic) in the samples was measured by cold vapour atomic absorption . FINDINGS: Mean mercury doses in infants exposed to thiomersal were 45.6 microg (range 37.5-62.5) for 2-month-olds and 111.3 microg (range 87.5-175.0) for 6-month-olds . Blood mercury in thiomersal-exposed 2-month-olds ranged from less than 3.75 to 20.55 nmol/L (parts per billion); in 6-month-olds all values were lower than 7.50 nmol/L . Only one of 15 blood samples from controls contained quantifiable mercury . Concentrations of mercury were low in urine after vaccination but were high in stools of thiomersal-exposed 2-month-olds (mean 82 ng/g dry weight) and in 6-month-olds (mean 58 ng/g dry weight) . Estimated blood half-life of ethylmercury was 7 days (95% CI 4-10 days) . INTERPRETATION: Administration of vaccines containing thiomersal does not seem to raise blood concentrations of mercury above safe values in infants . Ethylmercury seems to be eliminated from blood rapidly via the stools after parenteral administration of thiomersal in vaccines.

Infection, 2002 Dec, 30(6), 378 - 86
Efficacy and tolerability of once-daily telithromycin compared with high-dose amoxicillin for treatment of community-acquired pneumonia; Hagberg L et al.; BACKGROUND: This randomized, double-blind study compared the efficacy and tolerability of the new ketolide antimicrobial telithromycin with that of high-dose amoxicillin in the treatment of community-acquired pneumonia (CAP) . PATIENTS AND METHODS: Adult patients (n = 404), with signs and symptoms of CAP and radiologic confirmation were randomized to receive telithromycin 800 mg once daily (n = 199) or amoxicillin 1,000 mg three times a day (n = 205) for 10 days . Clinical and bacteriologic outcomes were assessed at post-therapy test-of-cure (days 17-24) and late post therapy (days 31-36) . RESULTS: The clinical cure rate for telithromycin-treated patients (per protocol) pst therapy (days 17-24) was 141/149 (94.6%) and compared well with that for amoxicillin (137/152 (90.1%)) . Subset analysis of patients (per protocol) showed high clinical cure rates for patients aged >/= 65 years (telithromycin 21/24, 87.5%; amoxicillin 22/29, 75.9%); those with documented pneumococcal bacteremia (telithromycin 10/10, 100%; amoxicillin 7/9, 77.8%); and patients with a Fine score >/= III (telithromycin 31/34, 91.2%; amoxicillin 38/47, 80.9%) . Bacterial eradication rates were comparable between treatments (telithromycin 42/48, 87.5%; amoxicillin 39/45, 86.7%), with 22/23 vs 18/21 Streptococcus pneumoniae strains 9/12 vs 11/13 Haemophilus influenzae strains and all Moraxella catarrhalis isolates (five and three patients, respectively) eradicated at the test-of-cure visit . Both treatments were generally well tolerated . CONCLUSION: Telithromycin 800 mg once daily is a convenient, optimal-spectrum, first-line treatment for CAP in adults, at least as effective and well tolerated as high-dose amoxicillin.

Pathobiology, 2002-2003, 70(2), 98 - 102
Elution of IgA from kidney tissues exhibiting glomerular IgA deposition and analysis of antibody specificity; Ogawa Y et al.; Glomerular IgA deposits were eluted from renal biopsy specimens exhibiting IgA nephropathy (IgAN) by using a combination of citrate buffer and collagenase . Collagenase predigestion of the kidney tissues resulted in increased amounts of IgA eluted by citrate buffer, and the elusion procedure did not attenuate the antigen-binding ability of IgA antibody . When reactivity of the eluted IgA with bacteria components was examined by Western blotting, the most notable reaction was observed for Haemophilus influenzae lysates in the form of a 34 kD-band . The reactivity of IgA eluted from the kidney tissues against the H . influenzae 34 kD antigen was evident in 3 of 5 IgAN cases . However, similar reactivity was also evident in 2 of 6 non-IgAN hepatic diseases exhibiting a glomerular IgA deposition . These findings suggest that antibody specificity of IgA against H . influenzae itself may not be directly associated with glomerular injury, although anti-H . influenzae 34 kD IgA was deposited in the kidney, at least in part, by IgAN . Further investigations into the properties of IgA deposited in the glomerulus are needed . Our improved method for IgA elution from kidney tissues would be useful for analysing the pathogenesis of IgAN .

Microb Pathog, 2002 Nov, 33(5), 211 - 23
Signature Tagged Mutagenesis of Haemophilus influenzae identifies genes required for in vivo survival; Herbert MA et al.; The pathogenic bacterium Haemophilus influenzae causes meningitis, epiglottitis, pneumonia, otitis media and other infections . To further understand the genetic basis of invasive disease and to inform about the bacterium's requirements in an in vivo environment, we analysed a library of 1632 insertional Tn1545 -Delta3 transposon mutants for their capacity to cause systemic infection in an animal model . We identified 25 genes that are potentially essential for H . influenzae invasive disease, and are candidates for further exploratory research . Seven of the genes encode hypothetical proteins, the function of six of which could be tentatively assigned on the basis of functional motifs and low homology to other bacterial genes . Eleven genes encode central metabolic enzymes or transporters; eight encode proteins that interact with DNA or modify other proteins; and four encode enzymes involved in the elaboration of classical virulence determinants . Two genes have no known function . Independent mutagenesis of six of the 25 genes and determination of the competitive index confirmed that these genes are important or essential to the organism in an in vivo environment . This genome-wide analysis has identified metabolic and other genes required during invasive disease, and the findings may lead to new interventions to prevent and treat H . influenzae infections.

Biochem Biophys Res Commun, 2002 Dec 20, 299(5), 710 - 4
Solution structure of the ribosome-associated cold shock response protein Yfia of Escherichia coli; Rak A et al.; The solution structure of the ribosome-associated cold shock response protein Yfia of Escherichia coli was determined by nuclear magnetic resonance with a RMSD of 0.6A . Yfia shows a global beta-alpha-beta-beta-beta-alpha folding topology similar to its homologue HI0257 of Haemophilus influenzae and the double-strand-binding domain of Drosophila Staufen protein . Yfia and HI0257 differ in their surface charges and in the composition of their flexible C-termini, indicating their specificity to different target molecules . Both proteins exhibit a hydrophobic and polar region, which probably functions as interaction site for protein complex formation . Despite their similarity to the dsRBD fold, Yfia does not bind to model fragments of 16S ribosomal RNA as determined by NMR titration and gel shift experiments.

J Orthop Surg (Hong Kong), 2001 Jun, 9(1), 83 - 90
Review article: Paediatric bone and joint infection; Stott NS; Paediatric musculoskeletal infection remains an important cause of morbidity . Methicillin sensitive Staphylococcus aureus is still the most common organism although the incidence of methicillin resistant S . aureus in the community is rising . Osteomyelitis and septic arthritis due to Haemophilus influenzae is decreasing in incidence secondary to immunisation and in some units has been replaced by infections with the gram negative bacillus, Kingella kingae . Recent prospective studies indicate that uncomplicated osteomyelitis can be treated by three to four weeks of antibiotics . However, there is still a small group of children who will have overwhelming disseminated infection . These children require aggressive surgical and medical intervention . Two recent reports have identified an increased incidence of septic arthritis in children who have hemophilia and are HIV positive.

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