Streptococcal

Chemotherapy, 2004 Jun, 50(3), 107 - 12
Pharmacodynamics and bactericidal activity of gatifloxacin in experimental pneumonia caused by penicillin-resistant Streptococcus pneumoniae; Yanagihara K et al.; BACKGROUND: Antimicrobial resistance rates for Streptococcus pneumoniae continue to increase worldwide, and resistance to nearly every major class of antimicrobials used to treat pneumococcal infections has been reported . Gatifloxacin (GFLX) is one of the quinolones that have strong activity against S . pneumoniae . METHODS: We compared the bacteriological, pharmacological and histopathological effects of orally administered GFLX with those of levofloxacin (LVFX) and ciprofloxacin (CPFX) in a murine model of pneumonia caused by penicillin-resistant S . pneumoniae (PRSP) . RESULTS: Treatment with GFLX resulted in a significant decrease in the number of viable bacteria (control, CPFX, LVFX, and GFLX: 6.48 +/- 0.36, 6.44 +/- 0.27, 5.51 +/- 0.15, and 4.89 +/- 0.28 log10 CFU/lung, respectively, mean +/- SD) . A significant decrease in mortality was observed in the GFLX-treated group in comparison with the other groups . Histopathological examination revealed that inflammatory changes in GFLX-treated mice were less marked than in the other mice . CONCLUSION: Our results suggest that orally administered GFLX is effective in PRSP pneumonia . The pharmacokinetic profiles also reflected the effectiveness of GFLX.

Proc Natl Acad Sci U S A, 2004 Aug 10, 101(32), 11833 - 8 Epub 2004 Jul 28.
Genome-wide molecular dissection of serotype M3 group A Streptococcus strains causing two epidemics of invasive infections; Beres SB et al.; Molecular factors that contribute to the emergence of new virulent bacterial subclones and epidemics are poorly understood . We hypothesized that analysis of a population-based strain sample of serotype M3 group A Streptococcus (GAS) recovered from patients with invasive infection by using genome-wide investigative methods would provide new insight into this fundamental infectious disease problem . Serotype M3 GAS strains (n = 255) cultured from patients in Ontario, Canada, over 11 years and representing two distinct infection peaks were studied . Genetic diversity was indexed by pulsed-field gel electrophoresis, DNA-DNA microarray, whole-genome PCR scanning, prophage genotyping, targeted gene sequencing, and single-nucleotide polymorphism genotyping . All variation in gene content was attributable to acquisition or loss of prophages, a molecular process that generated unique combinations of proven or putative virulence genes . Distinct serotype M3 genotypes experienced rapid population expansion and caused infections that differed significantly in character and severity . Molecular genetic analysis, combined with immunologic studies, implicated a 4-aa duplication in the extreme N terminus of M protein as a factor contributing to an epidemic wave of serotype M3 invasive infections . This finding has implications for GAS vaccine research . Genome-wide analysis of population-based strain samples cultured from clinically well defined patients is crucial for understanding the molecular events underlying bacterial epidemics.

Oper Dent, 2004 Jul-Aug, 29(4), 369 - 75
In vivo antibacterial effects of dentin primer incorporating MDPB; Imazato S et al.; This study examined the hypothesis that experimental primer containing the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB), which was previously reported to show bactericidal effects in vitro, inhibits bacteria in cavities under in vivo conditions . The number of bacteria resulting from applying primer solution to cavities in dog teeth infected with Streptococcus mutans was determined . The infected cavities were also restored using primer and the pulp response was histopathologically examined after 7, 30 and 75 days . No bacteria were recovered after applying the experimental primer, although the bactericidal effects of the proprietary primer were insignificant . Restoration with the experimental primer resulted in little or no pulpal inflammation for all periods; whereas, mild to moderate inflammatory response was observed when using proprietary primer . These results indicate that the experimental primer containing MDPB could exhibit in vivo antibacterial effects, suggesting its possible clinical benefit.

Rev Port Cardiol, 2004 May, 23(5), 723 - 8
Viridans streptococcus endocarditis associated with spondylodiscitis; Luz A et al.; The authors report a case of a 78-year-old male, admitted to the Hospital with fever, lumbar pain and a systolic murmur . Viridans streptococcus endocarditis associated with spondylodiscitis was diagnosed . Images and results of the exams are presented . This case is compared with similar studies in the literature.

Rev Med Chil, 2004 May, 132(5), 588 - 94
{Bacteremic pneumococcal pneumonia in 45 hospitalized adults}; Rioseco ML et al.; BACKGROUND: The presence of bacteremia during a pneumococcal pneumonia is a sign of bad prognosis . AIM: To report a clinical experience with bacteremic pneumococcal pneumonia . PATIENTS AND METHODS: We reviewed the clinical and laboratory data from 45 adults (36 male, aged 17 to 97 years) with community acquired pneumonia (CAP) and Streptococcus pneumoniae bacteremia, hospitalized between January 1997 and August 2002 at the Puerto Montt Hospital (Southern Chile) . RESULTS: Eighty four percent of patients bad underlying aggravating conditions, mainly alcoholism (40%), chronic obstructive lung disease (17.8%) and renal failure (17.8%) . Seven percent were homeless . Fever, cough, dyspnea and sputum were the most common presenting symptoms . Five patients had pleural involvement . Four strains (8.9%) of S . pneumoniae had diminished susceptibility to penicillin . Nine patients died (case-fatality rate of 20%), but mortality was attributed to pneumonia in only three of them . Main factors associated with a higher mortality were renal failure, absence of cough, an arterial pH < 7.3 on admission, ICU hospitalization, shock, mechanical ventilation and an APACHE score > 16 . CONCLUSIONS: The high death rate of these patients could be explained mainly by underlying conditions . ICU management and higher cost preventive measures could reduce this rate.

Rev Med Chil, 2004 May, 132(5), 533 - 8
{Assessment of two DNA extraction methods to amplify the pneumolysin gene (PLY) from blood culture samples of Streptococcus pneumoniae}; Hernandez C et al.; BACKGROUND: Streptococcus pneumoniae is a common etiologic agent of invasive respiratory infections among children under 5 years of age and older adults . Isolation rates of S . pneumoniae by traditional culture techniques are low . AIM: To study the sensitivity and specificity of two different DNA extraction methods to amplify the ply gene, applied to three different types of blood culture broths, experimentally inoculated with S . pneumoniae . MATERIAL AND METHODS: DNA was extracted from the cultures using an organic method or a technique that consists in dilution, washing with NaOH and concentration of the sample . This was followed by PCR amplification of a 355 pb fragment of the pneumolysin gene (ply) . RESULTS: The organic DNA extraction method inhibited the PCR reaction at all concentrations studied (0.6 to 10(6) colony forming units/mL) . Using the NaOH extraction, ply gene amplification was positive in all three blood culture broths, but only at concentrations of 10(3) colony forming units/mL, or higher . Using the same DNA extraction method, PCR was negative when the broths were inoculated with seven other related bacterial species, which results in a 100% specificity . CONCLUSIONS: Detection of S . pneumoniae by amplification of ply gene from blood cultures using the protocol of NaOH for DNA extraction is specific and provides results in a short lapse . However, the diagnostic sensitivity is not optimal, which limits its clinical use.

Pediatr Clin North Am, 2004 Aug, 51(4), 939 - 59, viii-ix
Diagnosis and management of bacterial infections in the neonate; Gerdes JS; Perinatally acquired bacterial neonatal sepsis is a low-incidence,high-risk disease . Although incidence of the most common etiology,group B Streptococcus, has been reduced by prophylactic strategies,neonatal sepsis has not been eradicated, and vigilance must remain high . Accurate diagnosis is difficult: signs and symptoms are hard to distinguish from other causes of neonatal distress, and definitive diagnostic tests are not available . The clinician must make a judgment call, considering the perinatal history, the constellation of signs and symptoms, and the results of existing diagnostic tests,before neonatal sepsis can diagnosed or excluded . With diagnosis,knowledge of the specific disease states and clinical algorithms for management aid in formulating a plan of treatment with antimicrobial agents and supportive care.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 3193 - 5
High prevalence of the ermB gene among erythromycin-resistant streptococcus pneumoniae isolates in Germany during the winter of 2000-2001 and in vitro activity of telithromycin; Kresken M et al.; Of 595 isolates of Streptococcus pneumoniae from outpatients with respiratory tract infections, collected from 17 microbiology laboratories, 14.1% were resistant to erythromycin . Eighty-three erythromycin-resistant isolates were genetically analyzed, 83.1% of which harbored the ermB gene . Only four isolates (4.8%) harbored the mefA gene . Telithromycin exhibited potent activity against all isolates.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 3169 - 71
In vitro activities of telithromycin, linezolid, and quinupristin-dalfopristin against Streptococcus pneumoniae with macrolide resistance due to ribosomal mutations; Farrell DJ et al.; To date, 86 of 7,746 macrolide-resistant Streptococcus pneumoniae isolates from 1999 to 2002 PROTEKT (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin) surveillance studies were negative for methylase and efflux mechanisms . Mutations in 23S rRNA or the genes encoding riboprotein L4 or L22 were found in 77 of 86 isolates . Six isolates were resistant to quinupristin-dalfopristin and two were resistant to linezolid, while telithromycin demonstrated good activities against all isolates.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 3016 - 23
Emergence of Streptococcus pneumoniae with very-high-level resistance to penicillin; Schrag SJ et al.; Penicillin resistance threatens the treatment of pneumococcal infections . We used sentinel hospital surveillance (1978 to 2001) and population-based surveillance (1995 to 2001) in seven states in the Active Bacterial Core surveillance of the Emerging Infections Program Network to document the emergence in the United States of invasive pneumococcal isolates with very-high-level penicillin resistance (MIC > or = 8 microg/ml) . Very-high-level penicillin resistance was first detected in 1995 in multiple pneumococcal serotypes in three regions of the United States . The prevalence increased from 0.56% (14 of 2,507) of isolates in 1995 to 0.87% in 2001 (P = 0.03), with peaks in 1996 and 2000 associated with epidemics in Georgia and Maryland . For a majority of the strains the MICs of amoxicillin (91%), cefuroxime (100%), and cefotaxime (68%), were > or =8 microg/ml and all were resistant to at least one other drug class . Pneumonia (50%) and bacteremia (36%) were the most common clinical presentations . Factors associated with very highly resistant infections included residence in Tennessee, age of <5 or > or =65 years, and resistance to at least three drug classes . Hospitalization and case fatality rates were not higher than those of other pneumococcal infection patients; length of hospital stay was longer, controlling for age . Among the strains from 2000 and 2001, 39% were related to Tennessee(23F)-4 and 35% were related to England(14-)9 . After the introduction of the pneumococcal conjugate vaccine, the incidence of highly penicillin resistant infections decreased by 50% among children <5 years of age . The emergence, clonality, and association of very-high-level penicillin resistance with multiple drug resistance requires further monitoring and highlights the need for novel agents active against the pneumococcus.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 2831 - 7
In vitro activities of ME1036 (CP5609), a novel parenteral carbapenem, against methicillin-resistant staphylococci; Kurazono M et al.; ME1036, formerly CP5609, is a novel parenteral carbapenem with a 7-acylated imidazo{5,1-b}thiazole-2-yl group directly attached to the carbapenem moiety of the C-2 position . The present study evaluated the in vitro activities of ME1036 against clinical isolates of gram-positive and gram-negative bacteria . ME1036 displayed broad activity against aerobic gram-positive and gram-negative bacteria . Unlike other marketed beta-lactam antibiotics, ME1036 maintained excellent activity against multiple-drug-resistant gram-positive bacteria, such as methicillin-resistant staphylococci and penicillin-resistant Streptococcus pneumoniae (PRSP) . The MICs of this compound at which 90% of isolates were inhibited were 2 microg/ml for methicillin-resistant Staphylococcus aureus (MRSA), 2 microg/ml for methicillin-resistant coagulase-negative staphylococci, and 0.031 microg/ml for PRSP . In time-kill studies with six strains of MRSA, ME1036 at four times the MIC caused a time-dependent decrease in the numbers of viable MRSA cells . The activity of ME1036 against MRSA is related to its high affinity for penicillin-binding protein 2a, for which the 50% inhibitory concentration of ME1036 was approximately 300-fold lower than that of imipenem . In conclusion, ME1036 demonstrated a broad antibacterial spectrum and high levels of activity in vitro against staphylococci, including beta-lactam-resistant strains.

Drugs Exp Clin Res, 2004, 30(2), 43 - 5
Pustular impetigo with good response to clarithromycin; Rallis E et al.; Impetigo is a contagious superficial pyogenic infection of the skin caused by Staphylococcus aureus and/or by group A Streptococcus . Two main clinical forms are recognized: bullous impetigo and non-bullous impetigo . We present an unusual case of pustular impetigo in a 35-year-old man . The pustules were localized symmetrically in the groin and the patient was successfully treated with clarithromycin . In bullous impetigo, exfoliative toxins produced by Staphylococcus aureus are accepted as the basis for the bulla formation just below the stratum granulosum . Although clarithromycin is considered to be a second-choice therapy for bullous impetigo, it was highly effective in our case.

J Infect Dis, 2004 Aug 15, 190(4), 739 - 47 Epub 2004 Jul 13.
Evolution of erythromycin-resistant Streptococcus pneumoniae from Asian countries that contains erm(B) and mef(A) genes; Ko KS et al.; To investigate the genetic characteristics of erythromycin-resistant Streptococcus pneumoniae in Asian countries, 110 pneumococcal isolates were analyzed by multilocus sequence typing (MLST) . MLST analysis showed that 2 clonal complexes--CC236 (Taiwan19F-14 clone) and CC81 (Spain23F-1 clone)--are the major lineages of erythromycin-resistant S . pneumoniae in the Asian region . Pneumococcal isolates containing both the erm(B) and the mef(A) genes are thought to have originated from the Taiwan19F-14 clone containing the mef(A) gene, after introduction of the erm(B) gene . Further evolution of this variant clone has generated resistant strains with different sequence types . Dissemination of these variant clones of the Taiwan19F-14 could be the main reason for the high frequency of pneumococcal isolates containing both erm(B) and mef(A) in some Asian countries . Data suggest that the high prevalence of erythromycin-resistant S . pneumoniae in the Asian region is partly due to the clonal spread of a few multidrug-resistant clones.

J Infect Dis, 2004 Aug 15, 190(4), 727 - 38 Epub 2004 Jul 20.
Progress toward characterization of the group A Streptococcus metagenome: complete genome sequence of a macrolide-resistant serotype M6 strain; Banks DJ et al.; We describe the genome sequence of a macrolide-resistant strain (MGAS10394) of serotype M6 group A Streptococcus (GAS) . The genome is 1,900,156 bp in length, and 8 prophage-like elements or remnants compose 12.4% of the chromosome . A 8.3-kb prophage remnant encodes the SpeA4 variant of streptococcal pyrogenic exotoxin A . The genome of strain MGAS10394 contains a chimeric genetic element composed of prophage genes and a transposon encoding the mefA gene conferring macrolide resistance . This chimeric element also has a gene encoding a novel surface-exposed protein (designated "R6 protein"), with an LPKTG cell-anchor motif located at the carboxyterminus . Surface expression of this protein was confirmed by flow cytometry . Humans with GAS pharyngitis caused by serotype M6 strains had antibody against the R6 protein present in convalescent, but not acute, serum samples . Our studies add to the theme that GAS prophage-encoded extracellular proteins contribute to host-pathogen interactions in a strain-specific fashion.

Infect Immun, 2004 Aug, 72(8), 4891 - 4
Inhibition of nitric oxide synthase exacerbates group B streptococcus sepsis and arthritis in mice; Puliti M et al.; The role of nitric oxide in group B Streptococcus (GBS) infection was evaluated by inhibiting its production with aminoguanidine (AG) . AG-treated mice displayed higher mortality rates and more frequent and severe arthritis than controls . Worsening of arthritis correlated with a higher number of GBS cells in the joints and local interleukin-1 beta production.

Infect Immun, 2004 Aug, 72(8), 4836 - 47
Proapoptotic effect of proteolytic activation of matrix metalloproteinases by Streptococcus pyogenes thiol proteinase (Streptococcus pyrogenic exotoxin B); Tamura F et al.; Streptococcus pyogenes thiol proteinase, also known as streptococcal pyrogenic exotoxin B (SpeB), has been suggested to be a major virulence factor in S . pyogenes infection . SpeB was reported to induce apoptosis of host cells, but its mechanism of action is not yet fully understood . In this study, we examined the involvement of matrix metalloproteinases (MMPs) in SpeB-induced apoptosis . We first developed a large-scale preparation of recombinant SpeB and precursors of human MMP-9 and -2 (proMMPs) by using Escherichia coli Rosetta (DE3)pLysS and baculovirus-insect cell expression systems, respectively . Treatment with SpeB induced effective proteolytic activation of both proMMP-9 and -2 . When RAW264 murine macrophages were incubated with SpeB-activated proMMP-9, the level of tumor necrosis factor alpha (TNF-alpha) in conditioned medium (CM), assessed by an enzyme immunoassay, was elevated . This increase was completely inhibited by addition of the MMP inhibitor SI-27 to the cell culture . The CM also produced marked induction of apoptosis of U937 human monocytic cells . Similarly, soluble Fas ligand (sFasL) was detected in CM of cultures of SW480 cells expressing FasL after treatment with SpeB-activated proMMPs; this CM also induced apoptosis in U937 cells . SpeB had a direct effect as well and caused the release of TNF-alpha and sFasL from the cells . SpeB-dependent production of MMP-9 and -2 and proapoptotic molecules (TNF-alpha and sFasL) was evident in a murine model of severe invasive S . pyogenes infection . These results suggest that SpeB or SpeB-activated MMPs contribute to tissue damage and streptococcal invasion in the host via extracellular release of TNF-alpha and sFasL.

Infect Immun, 2004 Aug, 72(8), 4680 - 8
Characterization of the Streptococcus mutans P1 epitope recognized by immunomodulatory monoclonal antibody 6-11A; Rhodin NR et al.; Monoclonal antibody (MAb) 6-11A directed against Streptococcus mutans surface adhesin P1 was shown previously to influence the mucosal immunogenicity of this organism in BALB/c mice . The specificity of anti-P1 serum immunoglobulin G (IgG) and secretory IgA antibodies and the subclass distribution of anti-P1 serum IgG antibodies were altered, and the ability of elicited serum antibodies to inhibit S . mutans adherence in vitro was in certain cases increased . MAb 6-11A is known to recognize an epitope dependent on the presence of the proline-rich region of the protein, although it does not bind directly to the isolated P-region domain . In this report, we show that MAb 6-11A recognizes a complex discontinuous epitope that requires the simultaneous presence of the alanine-rich repeat domain (A-region) and the P-region . Formation of the core epitope requires the interaction of these segments of P1 . Residues amino terminal to the A-region also contributed to recognition by MAb 6-11A but were not essential for binding . Characterization of the MAb 6-11A epitope will enable insight into potential mechanisms of immunomodulation and broaden our understanding of the tertiary structure of P1.

Infect Immun, 2004 Aug, 72(8), 4647 - 53
Attenuation of the bacterial load in blood by pretreatment with granulocyte-colony-stimulating factor protects rats from fatal outcome and brain damage during Streptococcus pneumoniae meningitis; Brandt CT et al.; A model of pneumococcal meningitis in young adult rats receiving antibiotics once the infection was established was developed . The intent was to mimic clinical and histopathological features of pneumococcal meningitis in humans . The primary aim of the present study was to evaluate whether medical boosting of the peripheral neutrophil count affected the outcome of the meningitis . The risk of terminal illness over the first 7 days after infection was significantly reduced for rats who had elevated peripheral white blood cell counts after receiving granulocyte-colony-stimulating factor (G-CSF) prior to the infection compared to that for untreated rats (P = 0.039 by the log rank test) . The improved outcome was associated with reduced signs of cerebral cortical damage (P = 0.008) . Furthermore, the beneficial effects of G-CSF were associated with reduced bacterial loads in the cerebrospinal fluid (median, 1.1 x 10(5) versus 2.9 x 10(5) CFU/ml; P = 0.023) and in blood (median, 2.9 x 10(2) versus 6.3 x 10(2) CFU/ml; P = 0.024), as well as attenuated pleocytosis (median, 800 x 10(6) versus 1,231 x 10(6) cells/liter; P = 0.025), 24 h after the infection . Conversely, initiation of G-CSF therapy 28 h postinfection did not alter the clinical or histological outcome relative to that for non-G-CSF-treated rats . The magnitude of bacteremia and pretreatment with G-CSF were found to be prognostic factors for both outcome and brain damage . In summary, elevated neutrophil levels prior to the development of meningitis result in reduced risks of death and brain damage . This beneficial effect is most likely achieved through improved control of the systemic disease.

Infect Immun, 2004 Aug, 72(8), 4579 - 88
phgABC, a three-gene operon required for growth of Streptococcus pneumoniae in hyperosmotic medium and in vivo; Brown JS et al.; To cause disease, bacterial pathogens need to be able to adapt to the physiological conditions found within the host, including an osmolality of approximately 290 mosmol kg(-1) . While investigating Streptococcus pneumoniae genes contained within pneumococcal pathogenicity island 1, we identified a three-gene operon of unknown function termed phgABC . PhgC has a domain with similarity to diacylglycerol kinases of eukaryotes and is the first described member of a family of related proteins found in many gram-positive bacteria . phgA and phgC mutant strains were constructed by insertional duplication mutagenesis and found to have impaired growth under conditions of high osmotic and oxidative stress . The compatible solutes proline and glycine betaine improved growth of the wild-type and the phgA mutant strains in hyperosmolar medium, and when analyzed by electron microscopy, the cellular morphology of the phgA mutant strain was unaffected by osmotic stress . The phgA and phgC mutant strains were reduced in virulence in models of both systemic and pulmonary infection . As the virulence of the phgA mutant strain was not restored in gp91phox(-/-) mice and the phgA and phgC mutant strains had reduced growth in both blood and serum, the reduced virulence of these strains is unlikely to be due to increased sensitivity to the respiratory burst of phagocytes but is, instead, due to impaired growth at physiological osmolality.

Infect Immun, 2004 Aug, 72(8), 4534 - 40
Protection against pneumococcal pneumonia in mice by monoclonal antibodies to pneumolysin; Garcia-Suarez Mdel M et al.; Pneumolysin (PLY) is an important virulence factor of Streptococcus pneumoniae . We examined the ability of three murine monoclonal antibodies (MAbs) to PLY (PLY-4, PLY-5, and PLY-7) to affect the course of pneumococcal pneumonia in mice . The intravenous administration of antibodies PLY-4 and PLY-7 protected the mice from the lethal effect of the purified toxin . Mice treated with PLY-4 before intranasal inoculation of S . pneumoniae type 2 survived longer (median survival time, 100 h) than did untreated animals (median survival time, 60 h) (P < 0.0001) . The median survival time for mice treated with a combination of PLY-4 and PLY-7 was 130 h, significantly longer than that for mice given isotype-matched indifferent MAbs (P = 0.0288) or nontreated mice (P = 0.0002) . The median survival time for mice treated with a combination of three MAbs was significantly longer (>480 h) than that for mice treated with PLY-5 (48 h; P < 0.0001), PLY-7 (78 h; P = 0.0007), or PLY-4 (100 h; P = 0.0443) alone . Similarly, the survival rate for mice treated with three MAbs (10 of 20 mice) was significantly higher than the survival rate obtained with PLY-5 (1 of 20; P = 0.0033), PLY-4 (2 of 20; P = 0.0138), or PLY-7 (3 of 20; P = 0.0407) alone . These results suggest that anti-PLY MAbs act with a synergistic effect . Furthermore, MAb administration was associated with a significant decrease in bacterial lung colonization and lower frequencies of bacteremia and tissue injury with respect to the results for the control groups.

Expert Rev Vaccines, 2004 Aug, 3(4), 375 - 8
Vaccines for other neonatal infections: vaccination strategies for the prevention of neonatal pertussis; McIntyre P; Current progress in the use of vaccines against a range of infectious agents, both bacterial (pertussis, pneumococcus and group B streptococcus) and mycobacterial (bacille Calmette-Guerin), to prevent neonatal infection are reviewed by Professors Gilbert, Britton and McIntyre and Dr Peter Richmond.

Expert Rev Vaccines, 2004 Aug, 3(4), 371 - 4
Vaccines for other neonatal infections: are group B streptococcal infections vaccine-preventable?
Gilbert GL.
Preliminary studies suggest that a pentavalent group B streptococcus conjugate vaccine, given in a single dose at 32-34 weeks gestation, would prevent approximately 90% of early and late onset neonatal and most postpartum maternal group B streptococcus infections.

Expert Rev Vaccines, 2004 Aug, 3(4), 365 - 9
Vaccines for other neonatal infections: neonatal BCG vaccination against tuberculosis; Britton WJ; This section focuses on current progress in the use of vaccines against a range of other infectious agents, both bacterial (pertussis, pneumococcus and group B streptococcus) and mycobacterial (bacille Calmette-Guerin) to prevent neonatal infection.

J Antimicrob Chemother, 2004 Aug, 54(2), 465 - 71 Epub 2004 Jul 21.
Effect of social and climatological factors on antimicrobial use and Streptococcus pneumoniae resistance in different provinces in Spain; Garcia-Rey C et al.; OBJECTIVES: To investigate the association between geographical differences in antibiotic consumption and resistance of Streptococcus pneumoniae to penicillin and erythromycin in 15 provinces of Spain, taking into account the potential influence of a series of social and climatological factors . METHODS: Possible correlations between prevalence of resistance to penicillin and erythromycin of S . pneumoniae, as determined in the national reference laboratory, and antibiotic consumption, and socio-economic and climatological variables were investigated . Partial correlations and multivariate linear regression were performed to assess the relative importance of variables predicting resistance and to investigate explicative factors for antibiotic consumption, respectively . RESULTS: A correlation was found between resistance and educational level, the proportion of young people in the population and climate, but was explained by their effects on differences in antibiotic use, which appeared to be the basic and only force behind resistance patterns in different geographical areas . Antibiotic use was found to be determined by the interplay of adult illiteracy, rainfall and GDP per capita . CONCLUSIONS: Interventions aimed at improving educational level and economic growth might therefore be followed by a noticeable reduction in overall antibiotic consumption, which might in turn be followed by a reduction in penicillin and erythromycin resistance in clinical isolates of S . pneumoniae.

Arch Dis Child, 2004 Aug, 89(8), 757 - 62
Twenty year surveillance of invasive pneumococcal disease in Nottingham: serogroups responsible and implications for immunisation; Ispahani P et al.; AIMS: To evaluate the incidence, spectrum of clinical manifestations, and outcome of invasive pneumococcal disease (IPD) in children . To determine the major serogroups of Streptococcus pneumoniae responsible for invasive disease and the potential coverage by the new pneumococcal conjugate vaccines . METHODS: Analysis of prospectively recorded information of all children admitted to two teaching hospitals in Nottingham with IPD between January 1980 and December 1999 . RESULTS: A total of 266 episodes of IPD in children were identified; 103 (39%) were aged <1 year and 160 (60%) <2 years . Major clinical presentations were meningitis in 86 (32%), pneumonia in 82 (31%), and bacteraemia without an obvious focus in 80 (30%) . The age specific mean annual incidence rates of IPD overall among children aged <1, <2, and <5 years were 47.1, 37.8, and 20 per 100 000 population, respectively . Mortality rates for children with meningitis and non-meningitic infection were 20% and 7%, respectively . Neurological sequelae following meningitis were documented in 16 (26%) of the 61 survivors assessed . The potential coverage rates in children between the ages of 6 months and 5 years are 84% by the 7-valent, 91% by the 9-valent, and 95% by the 11-valent conjugate vaccines . CONCLUSION: This study indicates that inclusion of a pneumococcal conjugate vaccine in the primary immunisation programme in the UK would have a considerable effect on the mortality and morbidity associated with IPD.

FEMS Microbiol Lett, 2004 Aug 1, 237(1), 57 - 64
Simple and rapid PCR method for identification of streptococcal species relevant to animal infections based on 23S rDNA sequence; Kawata K et al.; A PCR identification system targeting 23S rDNA sequences for the identification of eight streptococcal species relevant to animal infections (Streptococcus agalactiae, S . bovis, S . canis, S . dysgalactiae, S . equi, S . porcinus, S . suis and S . uberis) was developed . This system consists of two PCR reactions, A and B, in which seven and eight primers, respectively, are used simultaneously, and was designed so that each amplification product indicates a species by its size . A total of 111 cultures, including the type strain of eight species, could be successfully identified and differentiated as individual species, except for the cross reactivity between S . bovis and S . equinus . The developed PCR system can complete the identification procedure for eight streptococcal species through two tube reactions per isolate, and, therefore, might provide a rapid, simple and accurate diagnostic tool for veterinary laboratories.

Cochrane Database Syst Rev . 2004;(3):CD003520.
Vaginal chlorhexidine during labour to prevent early-onset neonatal group B streptococcal infection; Stade B et al.; BACKGROUND: Early-onset group B beta-hemolytic streptococcus (GBS) infection accounts for approximately 30% of neonatal infections, has a high mortality rate and is acquired through vertical transmission from colonized mothers . Several trials have demonstrated the efficacy of intrapartum chemoprophylaxis (IPC) for preventing early-onset disease (EOD) . Vaginal disinfection with chlorhexidine during labour has been proposed as another strategy for preventing GBS EOD in the preterm and term neonate . Chlorhexidine has been found to have no impact on antibiotic resistance, is inexpensive, and applicable to poorly equipped delivery sites . OBJECTIVES: To determine the effectiveness of vaginal disinfection with chlorhexidine during labour for preventing early-onset GBS infection in preterm and term neonates . SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth trials register (October 2003), the Cochrane Central Register of Controlled Trials (The Cochrane Library, Issue 3, 2003), MEDLINE (1966 to October 2003), EMBASE (1980 to March 2003), CINAHL (1982 to March 2003) and LILACS (1982 to September 2003) . SELECTION CRITERIA: Randomized and quasi-randomized trials comparing vaginal disinfection with chlorhexidine to placebo, or no treatment . DATA COLLECTION AND ANALYSIS: We extracted information from the results sections of the included studies . We reported relative risk (RR) and risk difference (RD) with 95% confidence intervals (CI) for dichotomous outcomes . We calculated the number needed to treat (NNT) with 95% CIs when a statistically significant RD was found . We used a chi square test (chi2) and the I2 analysis to test for heterogeneity, and applied a fixed or random effects model accordingly . MAIN RESULTS: Five studies, including approximately 2190 term and preterm infants, met the inclusion criteria and reported on at least one of the outcomes of interest for this systematic review . When all studies were combined there was a statistically significant (p = 0.005) reduction in colonisation (RR 0.72, 95% CI 0.56 to 0.91); RD -0.16 (95% CI -0.26 to -0.05); NNT 6 (95% CI 4 to 20) . There was no statistically significant between-study heterogeneity . There was no statistically significant between-study heterogeneity both for RR (chi(2) = 3.21 {p = 0.2}, I(2) = 37.8%) and for RD (chi(2) = 1.66 {p = 0.44}, I(2) = 0%) . There was no statistically significant reduction in EOD including GBS infection, GBS pneumonia, GBS meningitis or mortality . REVIEWERS' CONCLUSIONS: Vaginal chlorhexidine resulted in a statistically significant reduction in GBS colonisation of neonates, but was not associated with reductions in other outcomes . The review currently does not support the use of vaginal disinfection with chlorhexidine in labour for preventing EOD . Results should be interpreted with caution as the methodological quality of the studies was poor.

J Antimicrob Chemother, 2004 Aug, 54 Suppl 1, i23 - 9
Antibacterial susceptibility among Streptococcus pneumoniae isolated from paediatric and adult patients as part of the PROTEKT US study in 2001-2002; Brown SD et al.; BACKGROUND: One of the main factors commonly associated with antibacterial resistance among Streptococcus pneumoniae is the age of the patient . The highest rates of resistance have often been reported among isolates from young children . METHODS: Data from the PROTEKT US (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin in the United States) surveillance study were examined to determine the level of antibacterial resistance among S . pneumoniae isolates collected in 2001-2002 from different patient age groups in the USA . RESULTS: A total of 10 012 clinical isolates of S . pneumoniae were submitted by 242 centres across the USA and categorized into four patient age groups: infants (0-2 years, n = 1556), children (3-14 years, n=1125), adults (15-64 years, n=4058) and elderly adults (> or =65 years, n = 3067) (age unknown n=206) . With the exception of the fluoroquinolones and linezolid, rates of antibacterial resistance were highest among infants and decreased with increasing patient age . Resistance to penicillin ranged from 33.6% among infants to 17.5% among elderly adults, and erythromycin resistance ranged from 41.1% among infants to 24.0% among adults . In contrast, levofloxacin resistance increased with patient age (from 0.1% to 1.6%) . The highest rates of susceptibility were noted for telithromycin and linezolid (> or =99.6% and > or =99.8% susceptible isolates, respectively) . CONCLUSIONS: The PROTEKT US study data confirmed that the highest antibacterial resistance rates were associated with isolates collected from young children (0-2 years) . Telithromycin may offer a reliable alternative to first-line drugs in the empirical treatment of community-acquired respiratory tract infections.

J Antimicrob Chemother, 2004 Aug, 54 Suppl 1, i17 - 22
Distribution across the USA of macrolide resistance and macrolide resistance mechanisms among Streptococcus pneumoniae isolates collected from patients with respiratory tract infections: PROTEKT US 2001-2002; Farrell DJ et al.; BACKGROUND: Resistance to the macrolides has increased rapidly among isolates of Streptococcus pneumoniae from community-acquired respiratory tract infections (CARTIs) . METHODS: A total of 10 012 S . pneumoniae isolates were submitted from 46 US states and the territory of Puerto Rico in the PROTEKT US Year 2 (2001-2002) surveillance study . Antimicrobial susceptibilities were determined according to NCCLS guidelines and genes encoding common macrolide resistance mechanisms were sought by PCR . RESULTS: Overall, 27.9% (n=2793) of S . pneumoniae isolates were macrolide (erythromycin) resistant; the highest prevalence was recorded in Louisiana (48.2%) . Of the 2738 genotyped macrolide-resistant S . pneumoniae, 68.7% possessed mef(A) (state range: Delaware 40.0%-Georgia 84.8%), 16.8% of isolates harboured erm(B) (Georgia 6.1%-Idaho and Rhode Island both 36.4%) and 12.2% possessed erm(B) + mef(A) (Arkansas and Rhode Island 0%-South Dakota 32.9%) . Five isolates possessed an erm(A) subclass erm(TR) gene (from California, Illinois, Missouri, Pennsylvania and Virginia), while the mechanisms for 56 isolates were not definable by the methods used in this study . Susceptibility to telithromycin was high, irrespective of macrolide resistance mechanism, with > or =96.4% of the macrolide-resistant isolates susceptible . CONCLUSIONS: The prevalence of macrolide resistance and the resistance mechanisms among S . pneumoniae isolates are highly variable among the US states . Telithromycin may represent an effective treatment option for CARTIs caused by macrolide-resistant pneumococci.

Expert Opin Pharmacother, 2004 Aug, 5(8), 1701 - 10
Managing toxic shock syndrome with antibiotics; Annane D et al.; Toxic shock syndrome (TSS) is a serious disorder with a worldwide prevalence of approximately 3/100,000 persons . TSS is mainly caused by Streptococcus pyogenes or Staphylococcus aureus . Thus, beta-lactam and lincosamides, such as clindamycin, are the first-line drugs . Yet, the mortality rate remains unacceptably high; highlighting the role of bacterial toxin-mediated activation of the inflammatory cascade in TSS pathogenesis . Further strategies should be targeted towards interfering with the interaction between bacterial toxins and host T cells . This paper aims to provide an overview of the epidemiology, pathomechanisms, and clinical presentation of TSS, and criteria for selecting drugs among available antibiotics.

Expert Opin Pharmacother, 2004 Aug, 5(8), 1695 - 700
Treatment options in the management of necrotising fasciitis caused by Group A Streptococcus; Mulla ZD; Invasive Group A Streptococcus (GAS) disease is a serious condition that has multiple manifestations . A particularly severe form of invasive GAS disease is necrotising fasciitis (NF) . The case-fatality rate of GAS NF is approximately 20% . Penicillin remains the antibiotic of choice when treating invasive GAS infections . Epidemiological studies have shown that clindamycin is effective in the treatment of deep infections that are caused by GAS . Clinicians should consider adding clindamycin to the beta-lactam antibiotic regimen when NF or myositis is present . Intravenous immunoglobulin appears to be a promising adjunctive therapy in the management of GAS NF . Consultations with surgeons and infectious disease specialists are imperative.

Dis Aquat Organ, 2004 Jun 11, 59(3), 263 - 7
In vitro effect of a buffered chelating agent and neomycin or oxytetracycline on bacteria associated with diseases of fish; Wooley RE et al.; The antimicrobial agents used to treat bacterial fish diseases are archaic, and their uses may result in the emergence of drug-resistant bacterial strains . This study evaluated the in vitro antimicrobial activity of combinations of Tricide and neomycin or oxytetracycline on common disease-causing bacteria of fish and its possible use as an alternative treatment of these diseases . Tricide solutions containing of 8 mM United States Pharmacopeia (USP) disodium ethylenediaminetetraacetate dehydrate (chelator) and 20 mM USP 2-amino-2-hydroxymethyl-1,3-propanediol (buffer) potentate the antimicrobial action of neomycin and oxytetracycline when reacted in vitro with Aeromonas hydrophila, Streptococcus iniae, Pseudomonas aeruginosa, and Staphylococcus aureus . Serial passage of the test organisms in Tricide or Tricide and neomycin or oxytetracycline did not result in the development of resistant forms . Combinations of Tricide and neomycin or oxytetracycline reduced the amount of antibiotics necessary for fish therapy, render drug-resistant bacteria sensitive to antimicrobial therapy, may be used to decontaminate recently shipped fish, and should reduce the formation of antibiotic-resistant forms.

Rev Port Pneumol, 2003 Nov, 9(5 Suppl), 7 - 8
{Community-acquired pneumonia in a central hospital - comparison between a group of elderly and a non-elderly patients}; Moreira S et al.; The aim of our study was to compare CAP clinical attributes, etiologic agents and mortality between a group of elderly and a non-elderly patients . We performed a retrospective study analysing clinical data from 257 patients admitted to our hospital with CAP . Two groups were considered: group I with 65 years and over (111 patients) and group II with less then 65 years (146 patients) . Average age in group I was 77.3 years vs . 44.9 years in group II . Significant differences were found between two clinical characteristics: fever (54.1% in group I vs . 81.5% in group II) and chest pain (27.0% vs . 50.0% respectively) . Etiologic agents were identified in sputum cultures of 31.6% and in blood cultures of 6.2% of the patients in the group I vs 29.1% and 13.9% in the other group . Streptococcus pneumoniae was isolated in 8.8% of sputum cultures and in 3.1% of blood cultures of group I vs . 17.5% and 10.2% in group II . Enteric gram-negative bacteria were higher in group I than in group II (57.1% vs . 8.3%) . Duration of hospitalization (13.1 vs . 11.1 days) and mortality rate (10.8% vs . 0.7%) were higher for group I . In conclusion, the etiologic agents found were different in the two groups . The elderly patients had worst prognosis and required additional health care measures.

Proc Natl Acad Sci U S A, 2004 Jul 27, 101(30), 11123 - 8 Epub 2004 Jul 19.
Discovery and characterization of sialic acid O-acetylation in group B Streptococcus; Lewis AL et al.; Group B Streptococcus (GBS) is the leading cause of human neonatal sepsis and meningitis . The GBS capsular polysaccharide is a major virulence factor and the active principle of vaccines in phase II trials . All GBS capsules have a terminal alpha 2-3-linked sialic acid {N-acetylneuraminic acid (Neu5Ac)}, which interferes with complement-mediated killing . We show here that some of the Neu5Ac residues of the GBS type III capsule are O-acetylated at carbon position 7, 8, or 9, a major modification evidently missed in previous studies . Data are consistent with initial O-acetylation at position 7, and subsequent migration of the O-acetyl ester at positions 8 and 9 . O-acetylation was also present on several other GBS serotypes (Ia, Ib, II, V, and VI) . Deletion of the CMP-Neu5Ac synthase gene neuA by precise, in-frame allelic replacement gave intracellular accumulation of O-acetylated Neu5Ac, whereas overexpression markedly decreased O-acetylation . Given the known GBS Neu5Ac biosynthesis pathway, these data indicate that O-acetylation occurs on free Neu5Ac, competing with the CMP-Neu5Ac synthase . O-acetylation often generates immunogenic epitopes on bacterial capsular polysaccharides and can modulate human alternate pathway complement activation . Thus, our discovery has important implications for GBS pathogenicity, immunogenicity, and vaccine design.

J Exp Med, 2004 Jul 19, 200(2), 267 - 72
The scavenger receptor MARCO is required for lung defense against pneumococcal pneumonia and inhaled particles; Arredouani M et al.; Alveolar macrophages (AMs) express the class A scavenger receptor macrophage receptor with collagenous structure (MARCO), but its role in vivo in lung defense against bacteria and environmental particles has not been studied . We used MARCO-deficient mice to directly test the in vivo role of AM MARCO in innate defense against pneumococcal infection and environmental particles . In a murine model of pneumococcal pneumonia, MARCO(-/-) mice displayed an impaired ability to clear bacteria from the lungs, increased pulmonary inflammation and cytokine release, and diminished survival . In vitro binding of Streptococcus pneumoniae and in vivo uptake of unopsonized particles by MARCO(-/-) AMs were dramatically impaired . MARCO(-/-) mice treated with the "inert" environmental particle TiO(2) showed enhanced inflammation and chemokine expression, indicating that MARCO-mediated clearance of inert particles by AMs prevents inflammatory responses otherwise initiated by other lung cells . Our findings point to an important role of MARCO in mounting an efficient and appropriately regulated innate immune response against inhaled particles and airborne pathogens.

Clin Radiol, 2004 Aug, 59(8), 743 - 52
Inter-observer variation in the interpretation of chest radiographs for pneumonia in community-acquired lower respiratory tract infections; Hopstaken RM et al.; AIM: To assess inter-observer variation in the interpretation of chest radiographs of individuals with pneumonia versus those without pneumonia . MATERIALS AND METHODS: Chest radiographs of out-patients with a lower respiratory tract infection (LRTI) were assessed for the presence of infiltrates by radiologists from three local hospitals and were reassessed by one university hospital radiologist . Various measures of inter-observer agreement were calculated . RESULTS: The observed proportional agreement was 218 in 243 patients (89.7%) . Kappa was 0.53 (moderate agreement) with a 95% confidence interval of 0.37 to 0.69 . The observed positive agreement (59%) was much lower than for negative agreement (94%) . Kappa was considerably lower, if chronic obstructive pulmonary disease was present (kappa = 0.20) or Streptococcus pneumoniae (kappa = -0.29) was the infective agent . CONCLUSION: The overall inter-observer agreement adjusted for chance was moderate . Inter-observer agreement in cases with pneumonia was much worse than the agreement in negative (i.e . non-pneumonia) cases . A general practitioner's selection of patients with a higher chance of having pneumonia for chest radiography would thus not improve the observer agreement .

Quintessence Int, 2004 Jul-Aug, 35(7), 572 - 6
Naso-orbicular tissue necrosis by Streptococcus parasanguis in a patient with Fanconi anemia: clinical and laboratory aspects; Gomes MF et al.; Fanconi anemia (FA) is a rare autosomal recessive disorder, characterized by pancytopenia and progressive hypoplasia of the bone marrow . A 23-year-old woman with FA showed severe pancytopenia and developed an abscess on the infraorbicular region on the right side of the face that progressed to phlegmon and caused tissue necrosis of the nostrils, nasal septum, nasal fossa, and posterior orbital region . Laboratory examination showed Streptococcus parasanguis as the etiologic agent of the phlegmon . Supportive treatment was recommended due to donor incompatibility for bone marrow transplant . The intraoral examination showed spontaneous gingival bleeding, edema of the interdental papillae, hematomas on the superior and inferior lips, bacterial and fungal infections, and adequate oral hygiene . The patient was treated with the administration of an antibiotic (imipenem), an antifungal (amphotericin B), and mouth washing with antiseptic solutions . Periodontal prophylaxis and orientation to and control of oral hygiene and diet were also used during the remission period . For functional and esthetic rehabilitation of the alar regions and nasal dorsum, an acrylic resin nasal prosthesis was made, supported by a spectacle frame.

Folia Microbiol (Praha), 2004, 49(3), 307 - 14
Characterization of a complex restriction-modification system detected in Staphylococcus aureus and Streptococcus agalactiae strains isolated from infections of domestic animals; Godany A et al.; Characterization of classic type II restriction-modification systems (RMS) (restriction endonucleases and modification methyltransferases) was carried out in isolates of Staphylococcus aureus and Streptococcus agalactiae obtained from clinical material . Among the 100 isolates of S . aureus two different RMS type II were detected . The first was expressed in isolates 32 and 33 (Sau32 I and Sau33 I); the targeting sequence was determined as 5'-GGN CC-3' (Sau96 I isoschizomer) . The second was found in isolates no . 90, 93, 96*, and 98 (Sau90 I, Sau93 I, Sau96* I, Sau98 I) and enzymes recognized sequence 5'-CTY RAG-3' (SmlI isoschizomer) . Analysis of 40 isolates of S . agalactiae revealed only one RMS; it was detected in two isolates (no . 16 and 23; Sag16 I and Sag23 I) . Restriction endonuclease expressed by these isolates cleaved DNA in sequence 5'-CTG CA/G-3' (PstI isoschizomer) . In RMS-positive S . aureus and S . agalactiae isolates plasmid DNA capable of replication in Escherichia coli and Bacillus subtilis was also detected and isolated.

J Dairy Sci, 2004 Apr, 87(4), 813 - 5
Short communication: growth characteristics of Streptococcus uberis in UHT-treated milk; Dogan B et al.; Streptococcus uberis is an important environmental pathogen associated with bovine mastitis as well as with high total bacterial numbers in bulk tank milk . This study was conducted to determine whether S . uberis reproduction is likely to contribute to high bacterial numbers in bulk tank milk . Four S . uberis raw milk isolates were individually inoculated into UHT-treated milk and incubated at 4.4 or 7 degrees C for up to 5 d to simulate appropriate cooling; at 10 degrees C for 5 d to simulate marginally inadequate cooling; at 21 or 25 degrees C for 7 h to simulate ambient temperatures; or at 32 degrees C for 7 h to simulate elevated temperature conditions . None of the S . uberis isolates grew at either 4.4 or 7 degrees C . Streptococcus uberis growth at 10 degrees C appeared to be ribotype-specific . Although ribotype 116-520-S-1 isolates did not grow at 10 degrees C, ribotype 116-520-S-2 isolate numbers increased up to 3.5 log10 cfu/mL within 5 d . Generation times were calculated as 2.7 +/- 0.1 h, 2.1 +/- 0.1 h, and 1.0 +/- 0.1 h for 116-520-S-1 isolates and 1.8 +/- 0.4 h, 1.3 +/- 0.3 h, and 0.8 +/- 0.1 h for 116-520-S-2 isolates at 21, 25, and 32 degrees C, respectively . Our results suggest that high numbers of S . uberis in bulk tank milk are more likely to reflect high numbers of S . uberis shed by mastitic cows, rather than multiplication of these organisms under cooling conditions required for production of Grade A milk.

Eur J Clin Microbiol Infect Dis, 2004 Sep, 23(9), 665 - 71 Epub 2004 Jul 16.
Laboratory detection of group B Streptococcus for prevention of perinatal disease; Picard FJ et al.; Group B Streptococcus (GBS) or Streptococcus agalactiae emerged in the 1970s as the leading cause of neonatal morbidity and mortality . Today, GBS remains one of the leading causes of sepsis and meningitis in newborns despite important prevention efforts, including the issuance of recommendations for prevention of perinatal GBS disease by the American College of Obstetricians and Gynecologists, the Centers for Disease Control and Prevention, and the American Academy of Pediatrics in 1996/1997 . The gastrointestinal tract is the natural human reservoir for GBS and is the likely source of vaginal colonization . GBS disease in newborns usually results from ascending spread of GBS into the amniotic fluid, which leads to neonatal colonization and to invasive disease in some infants . This review analyzes the various laboratory methods available for the detection of GBS from clinical samples collected from pregnant women and will discuss their impact in the prevention of neonatal GBS infections and in the rationalization of antibiotic use . The recent commercial availability of a rapid and highly sensitive real-time polymerase chain reaction assay suitable for the specific detection of GBS from vagino-rectal samples obtained from pregnant women during delivery, which is approved by the US Food and Drug Administration, provides improvements in the accuracy and rapidity of GBS colonization screening compared to the standard culture-based method using the recommended selective enrichment broth .

Am J Epidemiol, 2004 Aug 1, 160(3), 270 - 8
Epidemiology of invasive Streptococcus pneumoniae among Navajo children in the era before use of conjugate pneumococcal vaccines, 1989-1996; O'Brien KL et al.; Streptococcus pneumoniae is the most common cause of invasive bacterial disease among children worldwide . The authors aimed to determine the incidence, clinical characteristics, and serotype distribution of invasive pneumococcal disease (IPD) among Navajo children in the southwestern United States . Active population-based laboratory surveillance for IPD among resident members of the Navajo Nation under 18 years of age was conducted between 1989 and 1996 . During this 8-year period, 706 cases of IPD were identified . The rate of disease varied by age, with the highest rate being observed among children aged 6-11 months (727 cases/100,000 person-years), followed by children aged 0-11 months, 0-23 months, and 0-59 months (568, 537, and 272 cases/100,000 person-years, respectively) . Among children aged 0-23 months, 60.3% of cases were caused by serotypes in the seven-valent conjugate pneumococcal vaccine (71.5% from 1989-1993 and 58.3% from 1994-1996) . Navajo children are at increased risk of IPD in comparison with the general US population . The distribution of disease-causing serotypes is similar to that of many countries in the developing world . Prevention strategies should include the use of licensed pneumococcal protein conjugate vaccine; however, a substantial proportion of disease is caused by nonvaccine serotypes . These data are critical for assessing the impact of these vaccines in this high-risk population.

Drugs, 2004, 64(15), 1683 - 94; discussion 1695-6
Telithromycin; Wellington K et al.; Telithromycin, the first member of the ketolide antibacterials, has good activity against community-acquired respiratory pathogens, including multiple-drug-resistant strains of Streptococcus pneumoniae . Telithromycin 800 mg once daily has been US FDA approved for the treatment of acute bacterial sinusitis (ABS; treatment duration 5 days), acute bacterial exacerbations of chronic bronchitis (AECB; 5 days) and mild-to-moderate community-acquired pneumonia (CAP; 7-10 days) . In patients with CAP, telithromycin was as effective as amoxicillin 1000 mg three times daily for 10 days, clarithromycin 500 mg twice daily for 10 days or trovafloxacin 200 mg once daily for 7-10 days . In patients with AECB, telithromycin was as effective as a 10-day regimen of amoxicillin/clavulanic acid 500/125 mg three times daily, cefuroxime axetil 500 mg twice daily or clarithromycin 500 mg twice daily . In patients with ABS, telithromycin was as effective as a 10-day course of amoxicillin/clavulanic acid 500/125 mg three times daily or cefuroxime axetil 250 mg twice daily . Telithromycin was generally well tolerated and most adverse events were of mild-to-moderate severity and transitory . The most common adverse events with telithromycin were diarrhoea and nausea (10.8% and 7.9% of 2702 patients in clinical trials); these events occurred in 8.6% and 4.6% of 2139 comparator-treated patients.

Presse Med, 2004 Jun 19, 33(11), 703 - 6
{Current Streptococcus pyogenes sensitivity responsible for acute tonsillopharyngitis in France}; Mariani-Kurkdjian P et al.; OBJECTIVE: Current guidelines recommend that only tonsillopharyngitis due to group A beta-haemolytic streptococcus (GABHS) diagnosed by rapid diagnostic test should be treated with antibiotics . Empirical antibiotic therapy must be based on epidemiological surveillance of resistance of GABHS to antibiotics . The aim of our study was to assess the activity of antimicrobial agents currently recommended for the treatment of GABHS tonsillopharyngitis . Method The activity of penicillin G, amoxicillin, cefaclor, cefpodoxime, cefuroxime, erythromycin, clarithromycin and clindamycin was determined against 93 consecutive GABHS isolates collected in 2002 . MIC50 and MIC90 of antibiotics tested were determined by agar dilution method according to CA-SFM guidelines . Macrolide resistance genes (ermA, ermB, mef) were detected by PCR . Genetic diversity of erythromycin-resistant isolates was analysed by pulsotypic method after digestion by SmaI (Finger-printing II, Biorad) . RESULTS: The activity of beta-lactam agents tested was similar and no resistant strain was detected (0%) . Nevertheless, this study shows an increasing emergence of erythromycin-resistant GABHS strains reaching 14% in 2002 (vs . 6.2% in a previous study carried out in 1996-1999) . CONCLUSION: The empirical antibiotic therapy of tonsillopharyngitis must consider, on the one hand, the high risk of GABHS eradication failure associated with in vitro resistance to erythromycin and clarithromycin, and on the other hand, the sustained susceptibility of GABHS to beta-lactam agents . These results reinforce the recommendations to use beta-lactam agents as first line treatment of GABHS tonsillopharyngitis.

Cornea, 2004 Aug, 23(6), 554 - 9
Outcome of combined Ahmed glaucoma valve implant and penetrating keratoplasty in refractory congenital glaucoma with corneal opacity; Al-Torbak AA; PURPOSE: To investigate surgical outcomes following simultaneous Ahmed glaucoma valve implant and penetrating keratoplasty (PKP) in the management of refractory congenital glaucoma with corneal opacity . METHODS: A retrospective review was undertaken of pediatric patients who underwent simultaneous Ahmed glaucoma valve implant and PKP at King Khaled Eye Specialist Hospital, Riyadh, Saudi Arabia, between January 1994 and September 1999 . RESULTS: Twenty eyes of 17 patients were included in the study . Cumulative probabilities of success by Kaplan-Meier analysis showed 85%, 44%, and 33% IOP control and 85%, 43%, and 17% graft success at 2, 24, and 48 months . The most common cause of glaucoma failure that required subsequent surgery was subconjunctival scarring, which resulted in loss of long-term IOP control . Main graft-related complications included failure (13/20) and graft ulceration (6/20), and in 4/6 ulcerated grafts, Streptococcus pneumoniae was cultured positively . Subsequent surgery was the only significant clinical factor associated with poor outcome of glaucoma . However, low graft survival rate was significantly associated with delinquency of follow-ups, corneal ulcer, subsequent surgery, and postoperative complications . CONCLUSIONS: The long-term success of simultaneous Ahmed glaucoma valve implant and PKP in refractory congenital glaucoma associated with corneal opacity is low, and the complication rate is high.

Cornea, 2004 Aug, 23(6), 547 - 9
Corneal infections after implantation of intracorneal ring segments; Hofling-Lima AL et al.; PURPOSE: To report risk factors, clinical course, and outcome in patients with infectious keratitis following implantation of intracorneal ring segments (ICRS) . METHODS: The records of 8 patients with culture-proven infectious keratitis after ICRS (Ferrara or Intacs) implantation were retrospectively reviewed . Age, gender, corneal findings, ocular abnormalities, the condition that led to ICRS implantation, immediate prior use of a contact lens, elapsed time between implantation and the onset of symptoms, previous medications, and systemic disorders were noted . RESULTS: Culture-positive infectious keratitis developed in 7 eyes of 7 patients (2 men and 5 women) with a mean age of 35 years who underwent Ferrara implantation for the treatment of keratoconus and in a 29-year-old man who underwent Intacs implantation for correction of low myopia . Contact lens use, diabetes, and trauma were factors possibly associated with the risk of infection in three cases . Microorganisms, identified in all cases, included Staphylococcus aureus, Streptococcus viridans, Streptococcus pneumoniae, Pseudomonas sp, Nocardia sp, Klebsiella sp, and Paecylomices sp . Onset of symptoms of infection varied from less than 1 week to 22 months postoperatively, depending on the infecting organism . CONCLUSIONS: Infectious keratitis following ICRS implantation is a sight-threatening complication for which early recognition and rapid institution of appropriate treatment may result in a better visual outcome.

Microbiology, 2004 Jul, 150(Pt 7), 2409 - 14
rexAB mutants in Streptococcus pneumoniae; Halpern D et al.; Streptococcus pneumoniae is a human pathogen that is naturally transformable . In this study a major component of the homologous recombination pathway, the RexAB exonuclease/helicase, was characterized . rexA and rexB insertional mutants were constructed using mariner mutagenesis and found to have identical phenotypes . Both rexAB mutants displayed poor cell viability, reduced double-strand exonuclease activity, UV sensitivity and a reduced level of gene conversion compared to the wild-type strain . No effect was observed on plasmid and chromosomal transformation efficiencies . These results indicate that in S . pneumoniae, RexAB is required for DNA repair, but not for chromosomal transformation and plasmid establishment.

Microbiology, 2004 Jul, 150(Pt 7), 2313 - 25
Relationship between codon biased genes, microarray expression values and physiological characteristics of Streptococcus pneumoniae; Martin-Galiano AJ et al.; A codon-profile strategy was used to predict gene expression levels in Streptococcus pneumoniae . Predicted highly expressed (PHE) genes included those encoding glycolytic and fermentative enzymes, sugar-conversion systems and carbohydrate-transporters . Additionally, some genes required for infection that are involved in oxidative metabolism and hydrogen peroxide production were PHE . Low expression values were predicted for genes encoding specific regulatory proteins like two-component systems and competence genes . Correspondence analysis localized 484 ORFs which shared a distinctive codon profile in the right horn . These genes had a mean G+C content (33.4 %) that was lower than the bulk of the genome coding sequences (39.7 %), suggesting that many of them were acquired by horizontal transfer . Half of these genes (242) were pseudogenes, ORFs shorter than 80 codons or without assigned function . The remaining genes included several virulence factors, such as capsular genes, iga, lytB, nanB, pspA, choline-binding proteins, and functions related to DNA acquisition, such as restriction-modification systems and comDE . In order to compare predicted translation rate with the relative amounts of mRNA for each gene, the codon adaptation index (CAI) values were compared with microarray fluorescence intensity values following hybridization of labelled RNA from laboratory-grown cultures . High mRNA amounts were observed in 32.5 % of PHE genes and in 64 % of the 25 genes with the highest CAI values . However, high relative amounts of RNA were also detected in 10.4 % of non-PHE genes, such as those encoding fatty acid metabolism enzymes and proteases, suggesting that their expression might also be regulated at the level of transcription or mRNA stability under the conditions tested . The effects of codon bias and mRNA amount on different gene groups in S . pneumoniae are discussed.

Mol Microbiol, 2004 Aug, 53(3), 889 - 901
Molecular analysis of the psa permease complex of Streptococcus pneumoniae; McAllister LJ et al.; The psaBCA locus of Streptococcus pneumoniae encodes a putative ABC Mn2+-permease complex . Downstream of the operon is psaD, which may be co-transcribed and encodes a thiol peroxidase . Previously, there has been discordance concerning the phenotypic impact of mutations in the psa locus, resolution of which has been complicated by differences in mutant construction and the possibility of polar effects . Here, we constructed unmarked, in frame deletion mutants DeltapsaB, DeltapsaC, DeltapsaA, DeltapsaD, DeltapsaBC, DeltapsaBCA and DeltapsaBCAD in S . pneumoniae D39 to examine the role of each gene within the locus in Mn2+ uptake, susceptibility to oxidative stress, virulence, nasopharyngeal colonization and chain morphology . The requirement for Mn2+ for growth and transformation was also investigated for all mutants . Inductively coupled plasma mass spectrometry (ICP-MS) analysis provided the first direct evidence that PsaBCA is indeed a Mn2+ transporter . However, this study did not substantiate previous reports that the locus plays a role in choline-binding protein pro-duction or chain morphology . We also confirmed the importance of the Psa permease in systemic virulence and resistance to superoxide and hydrogen peroxide, as well as demonstrating a role in nasopharyngeal colonization for the first time . Further evi-dence is provided to support the requirement for Mn2+ supplementation for growth and transformation of DeltapsaB, DeltapsaC, DeltapsaA, DeltapsaBC, DeltapsaBCA and DeltapsaBCAD mutants . However, transformation, as well as growth, of the DeltapsaD mutant was not dependent upon Mn2+ supplementation . We also show that, apart from sensitivity to hydrogen peroxide, the DeltapsaD mutant exhibited essentially similar phenotypes to those of the wild type . Western blot analysis with a PsaD antiserum showed that deleting any of the genes upstream of psaD did not affect its expression . However, we found that deleting psaB resulted in decreased expression of PsaA relative to that in D39, whereas deleting both psaB and psaC resulted in at least wild-type levels of PsaA .

FEMS Microbiol Lett, 2004 Jul 15, 236(2), 213 - 20
Mercuric resistance genes in gram-positive oral bacteria; Stapleton P et al.; Mercury-resistant bacteria isolated from the oral cavities of children carried one of two types of merA gene that appear to have evolved from a common ancestor . Streptococcus oralis, Streptococcus mitis and a few other species had merA genes that were very similar to merA of Bacillus cereus strain RC607 . Unlike the B . cereus RC607 merA gene, however, the streptococcal merA genes were not carried on Tn5084-like transposons . Instead, comparisons with microbial genomic sequences suggest the merA gene is located on a novel type II transposon . Coagulase-negative staphylococci and Streptococcus parasanguis had identical merA genes that represent a new merA variant.

FEMS Microbiol Lett, 2004 Jul 15, 236(2), 205 - 11
The streptococcolytic enzyme zoocin A is a penicillin-binding protein; Heath LS et al.; Zoocin A is a streptococcolytic enzyme produced by Streptococcus equi subsp . zooepidemicus 4881 that has an unknown site of action on the peptidoglycans of susceptible organisms . Analysis of a mutant strain in which the genes for zoocin A and resistance to zoocin A were inactivated revealed that this strain was more susceptible to beta-lactam antibiotics than the parental organism . Purified zoocin A had weak beta-lactamase activity, bound radioactive penicillin covalently, and its streptococcolytic activity was inhibited by penicillin . Thus, zoocin A is a penicillin-binding protein and presumably is a D-alanyl endopeptidase.

Semin Neonatol, 2004 Aug, 9(4), 289 - 302
Postmortem findings in term neonates; Pinar H; Neonatal deaths in infants born at term are relatively rare in the USA, occurring in 0.9/1000 live births . Congenital malformations, perinatal asphyxia, infections and inborn errors of metabolism are the leading causes . Chromosomal malformation syndromes, congenital heart disease, pulmonary hypoplasia and severe neural tube defects comprise the majority of lethal malformations . Several skeletal dysplasias are lethal in the newborn infant . Group B Streptococcus still plays a major role in neonatal mortality while deaths due to other infectious agents have decreased . Hypoxic ischaemic encephalopathy is a significant cause of neonatal death . Inborn errors of metabolism have variable presentations but some, such as the fatty acid oxidation disorders, may present in neonates and cause sudden death .

Arch Surg, 2004 Jul, 139(7), 760 - 5
Association of Streptococcus bovis bacteremia with colonic neoplasia and extracolonic malignancy; Gold JS et al.; BACKGROUND: The association between Streptococcus bovis bacteremia and colonic neoplasia is well described; however, the relationship between S bovis and neoplasia outside the colon has not been well evaluated . HYPOTHESIS: S bovis bacteremia may be associated with colonic neoplasia and extracolonic malignancy . DESIGN: Retrospective review of all documented cases of S bovis bacteremia identified by a search of computerized bacteriology records . SETTING: One tertiary referral hospital and 1 community hospital located in the same city . PATIENTS: Forty-five patients (41 adults, 4 children) with documented S bovis bacteremia during a 12-year period were identified . MAIN OUTCOME MEASURES: Available patient records were reviewed to identify the presence of colonic neoplasia, the use of gastrointestinal endoscopy, and the presence of gastrointestinal or extraintestinal malignancies . RESULTS: Seventeen patients (41% of adult patients) underwent colonoscopy . Colonic neoplasia was present in 16 patients (39% of adults), with 3 of these patients having invasive colorectal cancer (7% of adults) . Invasive cancer was present in 13 patients (32% of adults) . Eight patients had malignant lesions arising within the gastrointestinal tract, and 5 patients had extraintestinal malignancies . CONCLUSION: S bovis bacteremia is associated with both colonic neoplasia and extracolonic malignancy.

Biochemistry, 2004 Jul 20, 43(28), 9234 - 42
Characterization of the purified hyaluronan synthase from Streptococcus equisimilis; Tlapak-Simmons VL et al.; Hyaluronan synthase (HAS) utilizes UDP-GlcUA and UDP-GlcNAc in the presence of Mg(2+) to form the GAG hyaluronan (HA) . The purified HAS from Streptococcus equisimilis (seHAS) shows high fidelity in that it only polymerizes the native substrates, UDP-GlcNAc and UDP-GlcUA . However, other uridinyl nucleotides and UDP-sugars inhibited enzyme activity, including UDP-GalNAc, UDP-Glc, UDP-Gal, UDP-GalUA, UMP, UDP, and UTP . Purified seHAS was approximately 40% more active in 25 mM, compared to 50 mM, PO(4) in the presence of either 50 mM NaCl or KCl, and displayed a slight preference for KCl over NaCl . The pH profile was surprisingly broad, with an effective range of pH 6.5-11.5 and the optimum between pH 9 and 10 . SeHAS displayed two apparent pK(a) values at pH 6.6 and 11.8 . As the pH was increased from approximately 6.5, both K(m) and V(max) increased until pH approximately 10.5, above which the kinetic constants gradually declined . Nonetheless, the overall catalytic constant (120/s) was essentially unchanged from pH 6.5 to 10.5 . The enzyme is temperature labile, but more stable in the presence of substrate and cardiolipin . Purified seHAS requires exogenous cardiolipin for activity and is very sensitive to the fatty acyl composition of the phospholipid . The enzyme was inactive or highly activated by synthetic cardiolipins containing, respectively, C14:0 or C18:1(Delta9) fatty acids . The apparent E(act) for HA synthesis is 40 kJ (9.5 kcal/mol) disaccharide . Increasing the viscosity by increasing concentrations of PEG, ethylene glycol, glycerol, or sucrose inhibited seHAS activity . For PEGs, the extent of inhibition was proportional to their molecular mass . PEGs with average masses of 2.7, 11.7, and 20 kg/mol caused 50% inhibition of V(max) at 21, 6.5, and 3.5 mM, respectively . The apparent K(i) values for ethylene glycol, glycerol, and sucrose were, respectively, 4.5, 3.3, and 1.2 mM.

Int Microbiol, 2004 Jun, 7(2), 133 - 7
Peculiarities of the DNA of MM1, a temperate phage of Streptococcus pneumoniae; Obregon V et al.; The abundant presence of temperate phages in the chromosomes of clinical isolates of Streptococcus pneumoniae has been well documented . The genome of MM1, a temperate phage of pneumococcus, has been isolated as a DNA-protein complex . The protein is covalently bound to the DNA, was iodinated in vitro with Na125I, and has an Mr of 22,000 . Electron microscopy and enzymatic analyses revealed that the MM1 genome is a linear, circularly permuted, terminally redundant collection of double-stranded DNA molecules packaged via a headful mechanism . The location of the pac site appears to be downstream of the terminase, between orf32 and orf34 of the MM1 genome.

J Gen Appl Microbiol, 2004 Apr, 50(2), 71 - 8
Effects of the overexpression of fructose-1,6-bisphosphate aldolase on fermentation pattern and transcription of the genes encoding lactate dehydrogenase and pyruvate formate-lyase in a ruminal bacterium, Streptococcus bovis; Asanuma N et al.; Whether fructose-1,6-bisphosphate (FBP) triggers the transcriptional regulation of the gene expression of lactate dehydrogenase (LDH) and pyruvate formate-lyase (PFL) in Streptococcus bovis was examined by constructing a recombinant strain that overexpresses FBP aldolase (FBA) . When the recombinant strain was grown on glucose, intracellular FBP was much lower as compared to the parent strain, whereas dihydroxyacetone phosphate (DHAP) and d-glyceraldehyde-3-phosphate (GAP) were slightly higher . Intracellular ATP and ADP were slightly lower, but the NADH/NAD(+) ratio was not different . When glucose was replaced by lactose, a less readily utilized substrate, there was no great difference in FBP, DHAP, GAP, or adenine nucleotides . Overexpression of FBA decreased the level of LDH-mRNA, and increased the level of PFL-mRNA . Consequently, FBP concentration was positively related to the LDH-mRNA level and inversely related to the PFL-mRNA level . On the contrary, DHAP and GAP concentrations were positively related to the PFL-mRNA level and inversely related to the LDH-mRNA level . The levels of these mRNA were proportional to the amounts of corresponding enzymes in cells . As a result, the ratio of formate to lactate produced was increased by the overexpression of FBA . From these results, it could be presumed that FBP is involved in the transcriptional control of LDH and PFL synthesis in S . bovis.

Nat Med, 2004 Aug, 10(8), 811 - 3 Epub 2004 Jul 11.
A role for Streptococcus pneumoniae in virus-associated pneumonia; Madhi SA et al.; Here we show, in a double-blind, randomized, placebo-controlled trial in 37,107 fully immunized infants in Soweto, South Africa, that a 9-valent pneumococcal conjugate vaccine, PncCV, prevents 31% (95% confidence interval = 15-43%) of pneumonias associated with any of seven respiratory viruses in children in hospital . These data suggest that the pneumococcus has a major role in the development of pneumonia associated with these viruses and that viruses contribute to the pathogenesis of bacterial pneumonia.

Pediatr Infect Dis J, 2004 Jul, 23(7), 599 - 603
Trends in sepsis-related neonatal mortality in the United States, 1985-1998; Lukacs SL et al.; BACKGROUND: In the United States, bacterial sepsis affects up to 32,000 live births annually . In the 1990s, intrapartum antibiotic prophylaxis (IAP) was recommended to prevent maternal-infant transmission of group B Streptococcus (GBS), a leading cause of sepsis occurring in the first week of life (early onset sepsis) . Since IAP has been used, early onset GBS disease declined 70%; however, increased antibiotic use associated with IAP might lead to more severe or antimicrobial resistant etiologies of sepsis . To understand the influence of IAP on neonatal sepsis, in general, we evaluated neonatal mortality from sepsis before and after IAP recommendations were issued . METHODS: Using the National Center for Health Statistics Linked Birth/Infant Death Datasets, we compared trends in sepsis-related early neonatal mortality (<7 days) and late neonatal mortality (7-27 days) among singleton United States births from 1985 through 1991 to 1995 through 1998 {data beyond 1998 not included because of International Classification of Diseases (ICD)-10/ICD-9 coding differences} . We compared trends in mortality between the 2 time periods by estimating the average annual percent change in mortality using log linear regression and stratified by gestational age . RESULTS: Combined early and late neonatal mortality from sepsis averaged 39.6/100,000 live births from 1985 through 1991 and 31.8/100,000 live births from 1995 through 1998 . Early neonatal mortality from sepsis averaged 24.9/100,000 live births from 1985 through 1991 and 15.6 from 1995 through 1998; late neonatal mortality averaged 14.8/100,000 live births from 1985 through 1991 and 16.2 from 1995 through 1998 . Early neonatal mortality declined more steeply after IAP recommendations were issued, 5.0% annually from 1995 through 1998 versus 3.0% annually from 1985 through 1991 . Late neonatal mortality increased more from 1995 through 1998, 5.0% annually compared with 0.5% from 1985 through 1991 . CONCLUSIONS: Lower mortality rates and greater declines in early neonatal mortality from sepsis during 1995-1998 indicate greater survival of infants beyond 7 days of life and suggest an association with GBS disease prevention efforts . Thus these findings provide some evidence for continuing IAP for GBS-colonized women . Our findings of apparent increasing trends in late neonatal mortality from sepsis necessitate follow-up with clinical studies.

J Biol Chem, 2004 Sep 10, 279(37), 38571 - 6 Epub 2004 Jul 07.
Crystallographic and mutational data show that the streptococcal pyrogenic exotoxin J can use a common binding surface for T-cell receptor binding and dimerization; Baker HM et al.; The protein toxins known as superantigens (SAgs), which are expressed primarily by the pathogenic bacteria Staphylococcus aureus and Streptococcus pyogenes, are highly potent immunotoxins with the ability to cause serious human disease . These SAgs share a conserved fold but quite varied activities . In addition to their common role of cross-linking T-cell receptors (TCRs) and major histocompatibility complex class II (MHC-II) molecules, some SAgs can cross-link MHC-II, using diverse mechanisms . The crystal structure of the streptococcal superantigen streptococcal pyrogenic exotoxin J (SPE-J) has been solved at 1.75 A resolution (R = 0.209, R(free) = 0.240), both with and without bound Zn(2+) . The structure displays the canonical two-domain SAg fold and a zinc-binding site that is shared by a subset of other SAgs . Most importantly, in concentrated solution and in the crystal, SPE-J forms dimers . These dimers, which are present in two different crystal environments, form via the same face that is used for TCR binding in other SAgs . Site-directed mutagenesis shows that this face is also used for TCR binding SPE-J . We infer that SPE-J cross-links TCR and MHC-II as a monomer but that dimers may form on the antigen-presenting cell surface, cross-linking MHC-II and eliciting intracellular signaling.

J Biol Chem, 2004 Oct 15, 279(42), 43697 - 707 Epub 2004 Jul 09.
Structural and thermodynamic characterization of Pal, a phage natural chimeric lysin active against pneumococci; Varea J et al.; Pal amidase, encoded by pneumococcal bacteriophage Dp-1, represents one step beyond in the modular evolution of pneumococcal murein hydrolases . It exhibits the choline-binding module attaching pneumococcal lysins to the cell wall, but the catalytic module is different from those present in the amidases coded by the host or other pneumococcal phages . Pal is also an effective antimicrobial agent against Streptococcus pneumoniae that may constitute an alternative to antibiotic prophylaxis . The structural implications of Pal singular structure and their effect on the choline-amidase interactions have been examined by means of several techniques . Pal stability is maximum around pH 8.0 (Tm approximately 50.2 degrees C; DeltaHt = 183 +/- 4 kcal mol(-1)), and its constituting modules fold as two tight interacting cooperative units whose denaturation merges into a single process in the free amidase but may proceed as two well resolved events in the choline-bound state . Choline titration curves reflect low energy ligand-protein interactions and are compatible with two sets of sites . Choline binding strongly stabilizes the cell wall binding module, and the conformational stabilization is transmitted to the catalytic region . Moreover, the high proportion of aggregates formed by the unbound amidase together with choline preferential interaction with Pal dimers suggest the existence of marginally stable regions that would become stabilized through choline-protein interactions without significantly modifying Pal secondary structure . This structural rearrangement may underlie in vitro "conversion" of Pal from the low to the full activity form triggered by choline . The Pal catalytic module secondary structure could denote folding conservation within pneumococcal lytic amidases, but the number of functional choline binding sites is reduced (2-3 sites per monomer) when compared with pneumococcal LytA amidase (4-5 sites per monomer) and displays different intermodular interactions.

J Biol Chem, 2004 Sep 10, 279(37), 39017 - 25 Epub 2004 Jul 07.
High affinity streptococcal binding to human fibronectin requires specific recognition of sequential F1 modules; Schwarz-Linek U et al.; Fibronectin (Fn) binding by the Streptococcus pyogenes protein SfbI has been shown to trigger integrin-dependent internalization of this pathogen by human epithelial and endothelial cells . Here, using nuclear magnetic resonance spectroscopy and isothermal titration calorimetry in a dissection approach, the basis for the specificity and high affinity of the interaction between the N-terminal domain of Fn and SfbI is revealed . Each of the five Fn type 1 modules is directly involved in the interaction and is recognized by short consecutive motifs within the repeat region of SfbI . Crucially, these motifs must be combined in the correct order to form a high affinity ligand for the N-terminal domain of Fn.

Vaccine, 2004 Jul 29, 22(21-22), 2783 - 90
Identification and assessment of new vaccine candidates for group A streptococcal infections; McMillan DJ et al.; Group A Streptococcus (GAS) is a human-specific pathogen responsible for a wide variety of human diseases . Numerous GAS surface antigens interact with the human immune system and only some of these proteins have been studied in depth . A few of these may elicit protective response against GAS infection . In this study, we have used an in silico approach to identify antigenic peptides from GAS surface proteins . Putative GAS surface proteins from the M1 GAS genome were identified by the presence on LPxTG cell-wall anchoring motif and an export signal sequence . This technique identified 17 proteins of known or putative function, and another 11 which do not have known homologues . Peptides derived from predicted antigenic sequences near the amino terminus of six of these proteins, and another seven peptides derived from the two known surface proteins, GRAB and MtsA, were conjugated to keyhole lymphocyanin (KLH), and investigated for their capacity to induce opsonic antibody responses in outbred Quackenbush mice . All peptide-KLH antisera demonstrated opsonic capacity against both 88/30 and M1 GAS . However, KLH sera alone was also able to induce opsonic antibodies, suggesting that anti-KLH antibodies contributed to the opsonisation seen in the peptide-KLH antisera . KLH is therefore a promising carrier molecule for potential GAS peptide vaccines.

Diagn Microbiol Infect Dis, 2004 Jul, 49(3), 223 - 5
Clindamycin-resistant Streptococcus pyogenes: report of a case; Walsh SR et al.; A sentinel isolate of clindamycin-resistant Streptococcus pyogenes from a case of mixed aerobic-anaerobic necrotizing fasciitis prompted our clinical laboratory to change its protocol and subsequently perform routine susceptibility testing on all S . pyogenes isolated from blood and soft tissue specimens . Emerging clindamycin resistance may have serious implications in the treatment of severe S . pyogenes infections.

Cancer Sci, 2004 Jul, 95(7), 569 - 74
Frequent and preferential infection of Treponema denticola, Streptococcus mitis, and Streptococcus anginosus in esophageal cancers; Narikiyo M et al.; Multiple cancers frequently occur in the upper digestive tract . One possible explanation is that specific bacterial infection stimulates the normal epithelium to initiate inflammation and/or promotes carcinogenesis . This study was undertaken to determine which bacterial species is predominantly associated with esophageal cancer . We examined the bacterial diversity in this type of cancer and in the saliva from healthy people by using a culture-independent molecular method . Here we report the preferential and frequent infection of the oral periodontopathic spirochete Treponema denticola (T . denticola), Streptococcus mitis (S . mitis), and Streptococus anginosus (S . anginosus) in esophageal cancer from different regions of the world, and we also describe the induction of inflammatory cytokines by infection of S . anginosus and S . mitis . Our present data suggest that these three bacteria could have significant roles in the carcinogenic process of many cases of esophageal cancer by causing inflammation and by promoting the carcinogenic process, and that eradication of these three bacteria may decrease the risk of recurrence.

S Afr Med J, 2004 Jun, 94(6 Pt 2), 475 - 83
Guideline for the management of upper respiratory tract infections; Brink AJ et al.; INTRODUCTION: Inappropriate use of antibiotics for upper respiratory tract infections (URTIs), many of which are viral, adds to the burden of antibiotic resistance . Antibiotic resistance is increasing in Streptococcus pneumoniae, responsible for most cases of acute otitis media (AOM) and acute bacterial sinusitis (ABS) . METHOD: The Infectious Diseases Society of Southern Africa held a multidisciplinary meeting to draw up a national guideline for the management of URTIs . Background information reviewed included randomised controlled trials, existing URTI guidelines and local antibiotic susceptibility patterns . The initial document was drafted at the meeting . Subsequent drafts were circulated to members of the working group for modification . The guideline is a consensus document based upon the opinions of the working group . OUTPUT: Penicillin remains the drug of choice for tonsillopharyngitis . Single-dose parenteral administration of benzathine penicillin is effective, but many favour oral administration twice daily for 10 days . Amoxycillin remains the drug of choice for both AOM and ABS . A dose of 90 mg/ kg/day is recommended in general, which should be effective for pneumococci with high-level penicillin resistance (this is particularly likely in children < or = 2 years of age, in day-care attendees, in cases with prior AOM within the past 6 months, and in children who have received antibiotics within the last 3 months) . Alternative antibiotic choices are given in the guideline with recommendations for their specific indications . These antibiotics include amoxycillin-clavulanate, some cephalosporins, the macrolide/azalide and ketolide groups of agents and the respiratory fluoroquinolones . CONCLUSION: The guideline should assist rational antibiotic prescribing for URTIs . However, it should be updated when new information becomes available from randomised controlled trials and surveillance studies of local antibiotic susceptibility patterns.

J Clin Microbiol, 2004 Jul, 42(7), 3342 - 5
Differences in clinical manifestation of Streptococcus pneumoniae infection are not correlated with in vitro production and release of the virulence factors pneumolysin and lipoteichoic and teichoic acids; Spreer A et al.; Production and release of the pneumococcal virulence factors pneumolysin and lipoteichoic and teichoic acid in 75 clinical isolates were investigated . No difference was found between strains causing systemic infection or localized respiratory infection and isolates from asymptomatic carriers . This suggests that the presence of pneumolysin and lipoteichoic and teichoic acid is a necessary but not a sufficient condition for pneumococcal infection and development of invasive disease.

J Clin Microbiol, 2004 Jul, 42(7), 3303 - 5
Phenotypic and molecular characterization of erythromycin resistance in four isolates of Streptococcus-like gram-positive cocci causing bacteremia; Woo PC et al.; Among nine patients with bacteremia caused by Granulicatella or Gemella in a 6-year period (July 1995 to June 2001), three had bacteremia caused by erythromycin-resistant Granulicatella adiacens and one had bacteremia caused by erythromycin-resistant Gemella haemolysans . All four isolates possessed mef genes, whereas none possessed ermT, ermTR, or ermB genes.

J Clin Microbiol, 2004 Jul, 42(7), 3169 - 75
Multiplex PCR assay for detection of Streptococcus suis species and serotypes 2 and 1/2 in tonsils of live and dead pigs; Marois C et al.; A PCR assay was developed for the detection of Streptococcus suis serotypes 2 and 1/2 . This multiplex PCR is based on the amplification of the gene coding for 16S rRNA of S . suis and on the amplification of the cps2J gene coding for the capsule of S . suis serotypes 2 and 1/2 . An internal control was constructed and added in this test to monitor the efficiency of amplification in each reaction . To evaluate the specificity of the test, 31 strains of other bacterial species related to S . suis or isolated from pigs and 42 strains of S . suis serotypes 1 and 3 to 34 were analyzed . The detection threshold of the test was 28 S . suis CFU/ml . The specificity and the sensitivity of the multiplex PCR test and the presence of an internal control allowed the analysis of biological samples without a culture step . The PCR assay was then applied to the detection of 14 S . suis serotype 1/2 strains, 88 S . suis serotype 2 strains isolated from pigs, and 25 S . suis serotype 2 strains isolated from humans . This test was also applied to analyze tonsil samples of pigs experimentally infected and carrier pigs without any symptoms.

J Immunol, 2004 Jul 15, 173(2), 1307 - 12
Pneumolysin-induced lung injury is independent of leukocyte trafficking into the alveolar space; Maus UA et al.; Pneumolysin (PLY) is a major virulence factor released by Streptococcus pneumoniae and has been implicated in the pathogenesis of pneumococcal pneumonia . In this study, we evaluated the contribution of newly recruited neutrophils and monocytes and resident alveolar macrophages to the pathogenesis of PLY-induced lung injury . Mice received either adhesion-blocking Abs to inhibit alveolar leukocyte trafficking or liposomal clodronate to deplete alveolar macrophages before intratracheal application of native PLY or its noncytotoxic derivative PdB . We found that treatment with PLY but not PdB resulted in increased lung vascular permeability . In addition, PLY also induced a decrease in the resident alveolar macrophage population, and the recruitment of peripheral blood neutrophils and monocytes into the alveolar space . Blockade of PLY-induced alveolar leukocyte trafficking by pretreatment of mice with anti-CD18 plus anti-CD49d Abs or depletion of circulating neutrophils did not attenuate the increase in lung permeability observed in response to intratracheal PLY . In addition, depletion of resident alveolar macrophages with clodronated liposomes did not reduce alveolar injury developing in response to PLY . PLY-induced lung injury was associated with only a small increase in bronchoalveolar lavage concentrations of cytokines . These data indicate that PLY-induced lung injury results from direct pneumotoxic effects on the alveolar-capillary barrier and is independent of both resident and recruited phagocytic cells.

J Immunol, 2004 Jul 15, 173(2), 1194 - 201
Engagement of the pathogen survival response used by group A Streptococcus to avert destruction by innate host defense; Voyich JM et al.; Neutrophils are a critical component of human innate host defense and efficiently kill the vast majority of invading microorganisms . However, bacterial pathogens such as group A Streptococcus (GAS) successfully avert destruction by neutrophils to cause human infections . Relatively little is known about how pathogens detect components of the innate immune system to respond and survive within the host . In this study, we show that inactivation of a two-component gene regulatory system designated Ihk-Irr significantly attenuates streptococcal virulence in mouse models of soft tissue infection and bacteremia . Microarray analysis of wild-type and irr-negative mutant (irr mutant) GAS strains revealed that Ihk-Irr influenced expression of 20% of all transcripts in the pathogen genome . Notably, at least 11 genes involved in cell wall synthesis, turnover, and/or modification were down-regulated in the irr mutant strain . Compared with the wild-type strain, significantly more of the irr mutant strain was killed by human neutrophil components that destroy bacteria by targeting the cell envelope (cell wall and/or membrane) . Unexpectedly, expression of ihk and irr was dramatically increased in the wild-type strain exposed to these same neutrophil products under conditions that favored cell envelope damage . We report a GAS mechanism for detection of innate host defense that initiates the pathogen survival response, in which cell wall synthesis is critical . Importantly, our studies identify specific genes in the pathogen survival response as potential targets to control human infections.

Glycobiology, 2004 Nov, 14(11), 987 - 98 Epub 2004 Jul 07.
Trypanosome trans-sialidase targets TrkA tyrosine kinase receptor and induces receptor internalization and activation; Woronowicz A et al.; Trypanosome trans-sialidase (TS) is a sialic acid-transferring enzyme that hydrolyzes alpha2,3-linked sialic acids and transfers them to acceptor molecules . Here we show that a highly purified recombinant TS derived from T . cruzi parasites targets TrkA receptors on TrkA-expressing PC12 cells and colocalizes with TrkA internalization and phosphorylation (pTrkA) . Maackia amurensis lectin II (MAL-II) and Sambucus nigra lectin (SNA) block TS binding to TrkA-PC12 cells in a dose-dependent manner with subsequent inhibition of TS colocalization with pTrkA . Cells treated with lectins alone do not express pTrkA . The catalytically inactive mutant TSDeltaAsp98-Glu also binds to TrkA-expressing cells, but is unable to induce pTrkA . TrkA-PC12 cells treated with a purified recombinant alpha2,3-neuraminidase (Streptococcus pneumoniae) express pTrkA . Wild-type TS but not the mutant TSDeltaAsp98-Glu promotes neurite outgrowth in TrkA-expressing PC12 cells . In contrast, these effects are not observed in TrkA deficient PC12nnr5 cells but are reestablished in PC12nnr5 cells stably transfected with TrkA and are significantly blocked by inhibitors of tyrosine kinase (K-252a) and MAP/MEK protein kinase (PD98059) . Together these observations suggest for the first time that hydrolysis of sialyl alpha2,3-linked beta-galactosyl residues of TrkA receptors plays an important role in TrkA receptor activation, sufficient to promote cell differentiation (neurite outgrowth) independent of nerve growth factor.

J Exp Med, 2004 Jul 5, 200(1), 99 - 106
Bacterial inhibition of phosphatidylcholine synthesis triggers apoptosis in the brain; Zweigner J et al.; Streptococcus pneumoniae is the most common cause of bacterial meningitis of high mortality and morbidity . Neurological sequelae include paralysis, mental retardation, and learning disorders . In humans, neurons of the hippocampus undergo apoptosis as a result of meningitis . Phosphatidylcholine (PtdCho) is an essential component of mammalian cell membranes and PtdCho deficiency, either due to chemicals or altered nutrition, leads to apoptosis, especially in hippocampal neurons . We show that apoptosis of a variety of brain cells after pneumococcal infection arises from inhibition of PtdCho biosynthesis, the first such activity described for a bacterium . Apoptosis inhibitors did not prevent the bacterial-dependent inhibition of PtdCho biosynthesis . Supplementation with exogenous lyso-phosphatidylcholine prevents cell death and treatment of mice with cytidine diphosphocholine attenuates hippocampal damage during meningitis, even after the onset of infection . We conclude that bacterial inhibition of PtdCho biosynthesis activates an apoptotic cascade that is a causative event in pathogenesis and amenable to therapeutic intervention.

Arch Pediatr Adolesc Med, 2004 Jul, 158(7), 671 - 5
Incidence of occult bacteremia among highly febrile young children in the era of the pneumococcal conjugate vaccine: a study from a Children's Hospital Emergency Department and Urgent Care Center; Stoll ML et al.; BACKGROUND: The optimal diagnostic approach to and management of well-appearing, highly febrile young children has been a matter of debate owing to the possibility of clinically inapparent, or occult, bacteremia (OB) . The most common causative organism of OB is Streptococcus pneumoniae . Universal immunization with a heptavalent pneumococcal conjugate vaccine (PCV7) has recently been implemented, but there are limited data on the impact of this vaccine on the incidence of OB . OBJECTIVE: To evaluate the incidence of OB in the era of routine use of PCV7 . METHODS: We conducted a retrospective cohort study of highly febrile (temperature, 39 degrees C) children between the ages of 2 months and 36 months who had blood cultures performed in the emergency department or urgent care center between December 11, 2001, and March 5, 2003, and were discharged to home at the time of the initial visit . RESULTS: Of 329 blood cultures obtained from children who met inclusion criteria and did not meet exclusion criteria, 3 (0.91%; 95% confidence interval, 0%-1.9%) yielded a pathogenic bacterium; all were S pneumoniae . Neither an elevated total white blood cell count, an elevated absolute neutrophil count, nor an increased percentage of bands was highly predictive of OB . Blood cultures positive for organisms were more commonly due to contaminants (4; 95% confidence interval, 0%-2.4%) than pathogens . CONCLUSIONS: In the PCV7 era, OB is uncommon in highly febrile children 2 to 36 months of age . With continued use of PCV7, the routine practice of obtaining blood cultures and complete blood cell counts may no longer be indicated in previously healthy, well-appearing, highly febrile children 2 to 36 months of age, particularly those who have received at least 1 dose of PCV7.

J Infect, 2004 Aug, 49(2), 126 - 35
Genomic analysis of penicillin-resistant Streptococcus pneumoniae in Southeastern Michigan; Yee EK et al.; OBJECTIVE: The emergence of multidrug resistance within Streptococcus pneumoniae population was analysed, correlating penicillin resistance Pen(R) with secondary antibiotic resistance, capsular serotype, and genetic diversity among isolates . METHODS: DNA fingerprinting, following macro-restriction enzyme digestion and pulse field gel electrophoresis (PFGE), and restriction fragment analysis of the PBP 2b gene, following PCR amplification, were performed on the Pen(R) S . pneumoniae, among 377 clinical isolates obtained from the clinical microbiology laboratory (University of Michigan Medical Center) . RESULTS: Overall 35% of the isolates were Pen(R) of which 45% demonstrated high-level penicillin (Pen(R)-R, MIC>1) . Respiratory isolates were more likely to be Pen(R) (p <0.001) than non-respiratory isolates and the rate of Pen(R)-R was significantly increased in children <10 years of age (59.6%, p <0.02) . Secondary antibiotic resistance was more frequently associated with Pen(R)-R . Genomic DNA fingerprinting analysis and restriction fragment analysis of the PBP 2b gene demonstrated genomic divergence with discrete conserved pattern in the PBP 2b gene among the resistant isolates . CONCLUSION: The emergence of multidrug resistance in the S . pneumoniae population in SE Michigan is not due to expansion of a single or limited number of resistant clones, is occurring most frequently in the paediatric population and is associated with a decreased susceptibility to penicillin.

Laryngoscope, 2004 Jul, 114(7), 1147 - 50
Increasing antibiotic resistance of streptococcus species in New York City; Lin K et al.; OBJECTIVE: Streptococcus species are common pathogens in head and neck infections and are leading causes of morbidity and mortality . Emerging penicillin-resistant streptococcal pathogens have shifted empirical antibiotic therapy in favor of valuable alternatives, including erythromycin and clindamycin . This study was undertaken to determine the magnitude of antimicrobial resistance to these antibiotics . STUDY DESIGN: Retrospective review . METHODS: A retrospective study of two streptococcal species isolates, Streptococcus pyogenes (163 specimens) and Streptococcus pneumoniae (164 specimens), collected between January 1, 2001 and January 1, 2002 at two academic institutions . The antibiotic susceptibility patterns were analyzed for penicillin, erythromycin, and clindamycin according to the National Committee for Clinical Laboratory Standards . RESULTS: Fourteen percent to 34% of S . pyogenes isolates were erythromycin-resistant, and 0% to 28% were clindamycin-resistant . None of the S . pyogenes isolates were resistant to penicillin . Of the S . pneumoniae isolates, 33% to 50% were resistant to erythromycin, and 18% to 33% were resistant to clindamycin . The penicillin resistance levels for S . pneumoniae were 0% to 45% . CONCLUSIONS: Our antimicrobial resistance levels for S . pyogenes and S . pneumoniae significantly exceeded national and worldwide levels of erythromycin and clindamycin resistance . With a diverse population of over 8 million residents and high physician supply, our model is a microcosm for the study of antimicrobial use and susceptibility patterns and of clinical failure.

Rinsho Shinkeigaku, 2004 Mar, 44(3), 154 - 9
{An adult case of bacterial meningitis caused by penicillin-resistant streptococcus pneumoniae}; Igeta Y et al.; We report a patient of bacterial meningitis caused by penicillin-resistant streptococcus pneumoniae (PRSP) . A 50-year-old Japanese man was admitted after developing a fever and quickly falling into unconsciousness . Neurological examination showed slightly consciousness disturbance and meningeal irritation . A lumbar puncture yielded turbid spinal fluid, with increased cell count (411/mm3), protein (685 mg/dl) and IgG (60.3 mg/dl) but decreased glucose (1 mg/dl) . Bacterial meningitis was diagnosed and aminobenzylpenicillin (ABPC) and cefotaxime (CTX) were administered immediately, but they were ineffective . Penicillin-resistant streptcoccus pneumoniae (PRSP) was detected in the blood and spinal fluid, so antibiotics were changed to panipenem/betamipron (PAPM/BP) and vancomycin (VCM) with marked efficacy . With the increase in PRSP patients and documented failure in treatment of pneumococcal meningitis with ABPC and CTX, the need for alternative antibiotic therapy is critical . We emphasize the importance of initial therapy with PAPM/BP and VCM in patients with bacterial meningitis from streptcoccus pneumoniae.

Indian J Med Res, 2004 May, 119 Suppl, 213 - 20
Gene encoding the group B streptococcal protein R4, its presence in clinical reference laboratory isolates & R4 protein pepsin sensitivity; Smith BL et al.; BACKGROUND & OBJECTIVES: R proteins were first identified by Lancefield in group B Streptococcus (GBS) as resistant to trypsin at pH8 and sensitive to pepsin at pH2 . The R4 protein found predominantly in type III and some type II and V invasive isolates conforms to these criteria . The Rib protein, although structurally and epidemiologically similar to R4, was reported as resistant to both proteases . We report here the gene encoding the R4 protein from a type III group B streptococcal isolate (76-043) well characterized in our laboratory . METHODS: Trypsin extracted GBS proteins were assayed for protease sensitivities by double-diffusion Ouchterlony using varying conditions for the enzyme pepsin . Standard haemoglobin assay was used to examine pepsin enzymatic activity . Thirty clinical isolates of varying protein profiles identified by double-diffusion from our reference strain laboratory were screened by PCR and Southern technique . SDS-PAGE gel purified R4 amino acid sequences were determined and used to design oligonucleotide primers for screening a 76-043 genomic library . RESULTS: R4 was sensitive to pepsin at pH2 but appeared resistant at pH4, the reported pH used for Rib . By standard haemoglobin assay and trypsin extract studies of R4 protein, pepsin was shown to be active at pH2, yet easily inactivated; assays of GBS surface proteins are critical at pH2 . Of the amino acids initially sequenced from R4, 88 per cent (61/69) showed identity to Rib; the r4 nucleotide sequence was identical to that of rib . All isolates with strong positive protein reactions for R4 were positive in both PCR and Southern technique, whereas isolates expressing alpha, beta, R1/R4, and R5 (BPS) protein profiles were not . INTERPRETATION & CONCLUSION: Sequenced PCR products aligned with identity to the R4 and Rib nucleotide sequences and confirmed the identity of these proteins and their molecular sequences.

Indian J Med Res, 2004 May, 119 Suppl, 197 - 200
Towards a Belgian consensus for prevention of perinatal group B streptococcal disease; Melin P et al.; BACKGROUND & OBJECTIVES: In Belgium, as in many other countries, group B Streptococcus (GBS) is still the leading cause of sepsis and meningitis in neonates . In 2001, though no Belgian guidelines for their prevention were available, in some hospitals, obstetrical programmes included a GBS prevention policy . With an aim to reach a Belgian consensus for the prevention of perinatal group B streptococcal disease, a national consensum meeting was organized in 2001 . We report here our experience and findings of this meeting . METHODS: In November 2001, obstetricians, neonatologists, microbiologists and infectious diseases specialists were invited to participate in a GBS symposium . International and Belgian speakers presented epidemiological aspects, argued comparative cost-effectiveness of different approaches for prevention and debated technical and practical problems . Management of neonates with risk factors for GBS disease and progress in GBS vaccines were also included in the programme . Further results about Belgian obstetricians' practice and compliance to a policy for prevention of neonatal GBS diseases, as answered in two mail surveys, were commented and discussed . In an interactive session at the end, each participant was asked to vote on the key points related to the different steps of the ideal prevention strategy to recommend . RESULTS: For the main questions, 94 per cent of participants choose a screening-based approach and 94 per cent shifted from the current use of ampicillin to penicillin as first choice for antimicrobial prophylaxis . Further, 79 per cent voted for an approach with integrated neonatal prophylaxis for selected neonates at high risk for GBS disease and 47 per cent voted for a strategy based on an intrapartum rapid screening-based approach . INTERPRETATION & CONCLUSION: The state of the question by different speakers, the data from Belgian epidemiology, and the debate about cost-effectiveness of different approaches led to a massive vote in favour of the universal screening-based approach . Based on these results, a working group has been appointed by the Ministry of Health to draft and edit Belgian recommendations for the prevention of perinatal GBS disease.

Indian J Med Res, 2004 May, 119 Suppl, 179 - 82
Evaluation of gastrointestinal symptoms as primary sign of severe invasive group A streptococcal infections; Ekelund K et al.; BACKGROUND & OBJECTIVES: Severe invasive infections caused by group A Streptococcus (GAS) are often associated with shock and organ failure . We describe epidemiological and disease related data from the national surveillance of invasive GAS infection in Denmark in addition to three fatal cases that occurred in Denmark in 2002 with gastrointestinal (GI) symptoms as the dominating preliminary signs . METHODS: As the National Streptococcal Reference Centre The Streptococcus Unit, Statens Serum Institut (SSI) receives the vast majority of the invasive GAS isolates from patients admitted to all the hospitals in Denmark . The isolates were T-typed by slide agglutination test emm-squencing and pulsed field gel electrophoresis (PFGE) were also performed . RESULTS: During January 2002 three patients died at home and GAS were found at autopsy . Cases 1 (12 yr) and 3 (25 yr) had been ill for less than two days with nausea, diarrhoea and vomiting . Case 2 (25 yr) had the same symptoms for two weeks . None of the three had any underlying diseases . The GAS isolates from cases 1 and 2 were T-type 3-13-B3264, emm89 and SpeA-, SpeC- . The third isolate was Ttype 1, emm1 and SpeA+, SpeC- . PFGE could not discriminate between the two isolates with T-type 3-13-B3264 . The PFGE patterns of the three isolates were similar to those identified from GAS isolated elsewhere in Denmark at different times and from non-fatal cases . In 1999-2002, SSI received 409 isolates from patients with invasive GAS infection, and the mortality rate was 18 per cent . In 40 patients the primary symptoms were gastrointestinal, and in 30 per cent of these the outcome was fatal . INTERPRETATION & CONCLUSION: The various early clinical manifestations of severe GAS infections are still a major challenge for clinicians because of the importance of a fast and appropriate diagnosis and immediate start of treatment.

Indian J Med Res, 2004 May, 119 Suppl, 168 - 70
Comparison of Streptococcus pneumoniae serotypes causing acute otitis media & invasive disease in young children in the Czech Republic; Prymula R et al.; BACKGROUND & OBJECTIVES: The availability of a type-specific pneumococcal vaccine for children is a worldwide problem . It is necessary to study the serotypes prevalent in a country before introducing a type-specific vaccine . The objective of the present study was to analyse the prevalence of Streptococcus pneumoniae serotypes in children suffering from acute otitis media or invasive pneumococcal disease and to compare a coverage of serotypes by individual pneumococcal vaccines . METHODS: Children suffering from acute otitis media and invasive pneumococcal disease were analysed in the Czech Republic from October 1999 to November 2000 . Serotyping was performed by the quellung technique using antisera from Statens Serum Institute (Denmark) . RESULTS: The most frequent serotypes in patients with acute otitis media were 3, 19F, 23F, 14, 9V, 1, 6B, 11A and 28F . Vaccine coverage for the identified serotypes in acute otitis media patients was 52.1 per cent for the 7-valent vaccine, 57.8 per cent for the 9-valent vaccine and 75.7 per cent for the 11-valent form of the vaccine . In 108 patients with invasive pneumococcal disease, the most frequent serotypes were 6B, 9V, 14, 19F, 3 and 23F . Vaccine coverage for the identified serotypes in patients with invasive pneumococcal disease was 62 per cent for the 7-valent vaccine, 66.4 per cent for the 9-valent vaccine and 77.5 per cent for the 11-valent form of the vaccine . INTERPRETATION & CONCLUSION: Vaccine coverage for the identified serotypes for the 11-valent pneumococcal vaccine was better than the other two vaccines.

Indian J Med Res, 2004 May, 119 Suppl, 155 - 9
Epidemiology of diseases caused by Streptococcus pyogenes in Serbia during a nine-year period (1991-1999); Ranin L et al.; BACKGROUND & OBJECTIVES: Streptococcus pyogenes (group A Streptococcus - GAS) is an important human pathogen which causes a variety of diseases, including tonsillopharyngitis, scarlet fever and rheumatic fever . It is important to understand the changes in epidemiology of the diseases caused by the pathogen for improved control of such infections . Hence, the aim of the present study was to carry out an epidemiological analysis of GAS infections in Serbia in a 9-yr period (1991-1999) and evaluation of susceptibility of GAS isolates obtained during the same period to penicillin and erythromycin . METHODS: Occurrence of tonsillopharyngitis, scarlatina and rheumatic fever was analyzed and GAS carrier status in healthy children was examined over a 9-yr period from 1991 to 1999 . Susceptibility to penicillin and erythromycin was determined for 1657 GAS isolates obtained from patients diagnosed with pharyngitis or scarlet fever and 512 isolates from healthy carriers . M-type antigen was also determined in these isolates . RESULTS: The average incidences of tonsillopharyngitis and scarlet fever were 76.2 and 30.8 per cent respectively . A total of 166 cases of rheumatic fever were registered . Per cent of carriers varied from 5.5 to 11.4 per cent over the study period . Predominating M serotypes among GAS isolates tested were M1, M3, M4, M6, M11, M12 and M18, depending on the source of clinical material and period of isolation . Antimicrobial susceptibility testing showed susceptibility to penicillin in all isolates tested and resistance to erythromycin in 2.41 per cent of the isolates . INTERPRETATION & CONCLUSION: Although the fluctuations in incidence were noted during the nine-year period, the incidence of streptococcal tonsillopharyngitis is low but with a steady raise in Serbia . No significant changes in the incidence of scarlet fever and rheumatic fever were noted . Susceptibility to penicillin remained unchanged, but the number or erythromycin resistant strains have increased.

Indian J Med Res, 2004 May, 119 Suppl, 126 - 30
Antibody classes & subclasses induced by mucosal immunization of mice with Streptococcus pyogenes M6 protein & oligodeoxynucleotides containing CpG motifs; Teloni R et al.; BACKGROUND & OBJECTIVES: Type-specific antibodies against M protein are critical for human protection as they enhance phagocytosis and are protective . An ideal vaccine for the protection against Streptococcus pyogenes would warrant mucosal immunity, but mucosally administered M-protein has been shown to be poorly immunogenic in animals . We used a recombinant M type 6 protein to immunize mice in the presence of synthetic oligodeoxynucleotides containing CpG motifs (immunostimulatory sequences: ISS) or cholera toxin (CT) to explore its possible usage in a mucosal vaccine . METHODS: Mice were immunized by intranasal (in) or intradermal (id) administration with four doses at weekly intervals of M6-protein (10 microg/mouse) with or without adjuvant (ISS, 10 microg/mouse or CT, 0,5 microg/mouse) . M6 specific antibodies were measured by enzyme linked immunosorbent assay using class and subclass specific monoclonal antibodies . RESULTS: The use of ISS induced an impressive anti M-protein serum IgG response but when id administered was not detectable in the absence of adjuvant . When used in, M-protein in the presence of both ISS and CT induced anti M-protein IgA in the bronchoalveolar lavage, as well as specific IgG in the serum . IgG were able to react with serotype M6 strains of S . pyogenes . The level of antibodies obtained by immunizing mice in with M-protein and CT was higher in comparison to M-protein and ISS . The analysis of anti-M protein specific IgG subclasses showed high levels of IgG1, IgG2a and IgG2b, and low levels of IgG3 when ISS were used as adjuvant . Thus, in the presence of ISS, the ratio IgG2a/IgG1 and (IgG2a+IgG3)/IgG1 >1 indicated a type 1-like response obtained both in mucosally or systemically vaccinated mice . INTERPRETATION & CONCLUSION: Our study offers a reproducible model of anti-M protein vaccination that could be applied to test new antigenic formulations to induce an anti-group A Streptococcus (GAS) vaccination suitable for protection against the different diseases caused by this bacterium.

Indian J Med Res, 2004 May, 119 Suppl, 115 - 20
Immune responses of a liposome/ISCOM vaccine adjuvant against streptococcal fibronectin binding protein 1 (Sfb1) in mice; McArthur J et al.; BACKGROUND & OBJECTIVES: The fibronectin binding protein Sfb1 of Streptococcus pyogenes is a well characterised antigen which induces protection against lethal challenge with group A streptococcus (GAS) when adjuvanted with cholera toxin B-subunit (CTB) . As an alternative to CTB adjuvanted intranasal immunisations we investigated the immune responses generated in mice using Sfb1 incorporated in to the skin and mucosal adjuvant SAMA4 . METHODS: Mice (BALB/c) were vaccinated intradermally with 100 microl of either SAMA4 (adjuvant only group) or SAMA4/Sfb1 and were boosted 7 days later . Mice vaccinated with CTB based vaccines were immunised by intranasal inoculation with a mixture containing 30 microg Sfb1 and 10 microg CTB on days 1, 3, 5 and 15 . At 14 days after the last booster immunisation the immune response was characterised and mice were challenged with 10(8) CFU of S . pyogenes . RESULTS: Mice vaccinated with SAMA4/Sfb1 elicited a Sfb1-specific IgG response in the sera that was significantly higher than that seen in control mice and mice immunised with the adjuvant only (P<0.05) . No significant differences were seen for specific IgA antibodies in the sera in all groups examined . Compared with non-immunised and adjuvant only immunised controls, mice immunised with the Sfb1/SAMA4 vaccine exhibited a significant increase (P<0.05) in the number of Sfb1 reactive spleen cells in lymphoproliferation assays which were three fold higher than those seen for mice vaccinated with the Sfb1/CTB vaccine . Mice vaccinated with CTB/Sfb1 had the highest level of protection (80%) as where mice vaccinated with SAMA4 and SAMA4/Sfb1 displayed no protection (20% and 40%) . INTERPRETATION & CONCLUSION: These data suggest that the SAMA4 adjuvant used in this study fails to elicit protective immunity in BALB/c mice when used to adjuvant the known protective antigen Sfb1.

Indian J Med Res, 2004 May, 119 Suppl, 104 - 7
Preclinical evaluation of a vaccine based on conserved region of M protein that prevents group A streptococcal infection; Batzloff M et al.; BACKGROUND & OBJECTIVES: Infection with group A Streptococcus (GAS) may result in a number of human diseases ranging from the relatively benign pharyngitis to the potentially life-threatening invasive diseases and post-infectious sequelae . We have previously defined a minimal B-cell epitope from the conserved region of the M-protein . Here we report on the immunogenicity, opsonic potential of the resulting sera and the level of protection induced by this peptide in comparison to a pepsin extract of the M protein . METHODS: Inbred mice were immunized with peptides derived from the M protein . Sera were collected from the immunized mice and its opsonic potential determined for M1 and M6 GAS strains . Mice were then intranasally challenged with a virulent M1 GAS strain to determine the protective efficacy of the peptides . RESULTS: The peptides induced significant antibody responses when delivered subcutaneously and immunized mice demonstrated significantly enhanced survival compared to control groups following challenge . INTERPRETATION & CONCLUSION: The data obtained in the present study indicated that the chimeric peptide J8 from the conserved region of the M protein could form the basis for an anti-streptococcal vaccine in future.

Indian J Med Res, 2004 May, 119 Suppl, 99 - 103
Use of cDNA microarrays to analyze responses to pneumococcal virulence factors; McDaniel LS et al.; BACKGROUND & OBJECTIVES: The complex interactions that occur between host and pathogen during bacteraemia caused by Streptococcus pneumoniae are not well understood . Upon entering the blood stream the pneumococcus intiates responses through contact with naive monocytes and macrophages resulting in an inflammatory response . To elucidate the role of microbial virulence factors in the host response to the pneumococcus, cDNA microarray analysis was used to identify genes in THP-1 cells, a human monocytic cell line, that are responsive to pneumococcal virulence factors . METHODS: S . pneumoniae D39, a serotype 2 pneumococcus, and PLN an isogenic mutant of D39 that does not express pneumolysin were used . Gene expression profiles elicited by both wild-type and mutant were compared with that of THP-1 cells not exposed to pneumococci . Results obtained from microarray analysis were confirmed and further characterized using reverse transcriptase (RT)-PCR, real-time RT-PCR, and ELISA . RESULTS: Genes in THP-1 cells that were responsive to the pneumococcus independent of the presence of the specific virulence factor, pneumolysin, were identified . THP-1 cell genes that were differentially expressed independent of pneumolysin included the ones involved in cell-to-cell signaling and antipathogen responses . Those that were responsive to pneumolysin included genes encoding adhesion molecules, chemokines, cytokine receptors, and cell cycle and apoptosis proteins . INTERPRETATION & CONCLUSION: The global transcriptional response of naive monocytes to contact with the pneumococcus was characterized and the utility of cDNA microarray analysis in elucidating the role of specific factors in host-pathogen interactions were demonstrated.

Indian J Med Res, 2004 May, 119 Suppl, 84 - 7
Vaginal & rectal carriage of Streptococcus agalactiae in the Czech Republic: incidence, serotypes distribution & susceptibility to antibiotics; Motlova J et al.; BACKGROUND & OBJECTIVES: Streptococcus agalactiae (group B streptococcus, GBS) is the predominant bacterial agent responsible for invasive perinatal infection . To obtain reliable data on vaginal and rectal carriage of S . agalactiae in pregnant women in Czech Republic, and to formulate a prevention programme of neonatal GBS disease for the Czech Republic, women at childbirth were screened for vaginal and anorectal carriage of GBS . The isolates were serotyped and tested for susceptibility to antimicrobials including those recommended for intrapartum prophylaxis . METHODS: A total of 586 women at childbirth were screened for GBS carriage in vaginal and anorectal regions using the non-enrichment and selective culture media . The isolates were serotyped by precipitation with antisera raised against various serotypes and antigenic extracts prepared according to Lancefield's modification . Mueller Hinton agar with 5 per cent defibrinated sheep blood was used for antimicrobial susceptibility testing . MIC values were evaluated according to the NCCLS criteria . RESULTS: Using selective media, GBS was detected in 172 (29.3%) of 586 women screened, vaginal and anorectal colonization was found in 21.7 and 24.4 per cent of them, respectively, concomitant vaginal and anorectal colonization was recorded in 16.5 per cent of the women studied . Serotypes III (33.2%), Ia (22.0%) and V (13.9%) prevailed among 172 isolates tested . All isolates were susceptible to penicillin, ampicillin and cefotaxime . The rates of GBS resistance to tetracycline, erythromycin and clindamycin were 83.9, 3.8 and 3.2 per cent, respectively . INTERPRETATION & CONCLUSION: GBS carriage in pregnant women in the Czech Republic is rather high as compared with that reported in literature . The most frequent serotypes III, Ia and V, identified in GBS-colonized pregnant women in the Czech Republic, were among those predominant in the USA and Western Europe . Our findings confirm uniform susceptibility of GBS isolates from pregnant women to penicillin and other beta-lactam antibiotics tested . Resistance to erythromycin remains low in the Czech Republic.

Indian J Med Res, 2004 May, 119 Suppl, 66 - 73
Interaction of human factor H with PspC of Streptococcus pneumoniae; Dave S et al.; BACKGROUND & OBJECTIVES: Streptococcus pneumoniae has acquired virulence factors such as the polysaccharide capsule and various surface proteins, which prevent opsonization mediated by the complement system . PspC is one of the multi-functional pneumococcal surface proteins capable of eliciting an antibody response in mice . Our study further explores the role of pneumococcal surface proteins in resistance to complement mediated opsonophagocytosis by providing evidence that PspC binds human Factor H (FH), a regulatory protein of the alternative complement pathway . The present study was carried out to map the binding regions on PspC and FH, and to assess the functional activity of FH upon binding to PspC . METHODS: FH binding to D39 and other pneumococcal strains was observed by flow cytometry . A series of FH truncated and deletion mutants and PspC mutants were used to localize binding regions within these molecules . The functional activity of FH upon binding to PspC was measured by a haemolysis assay . RESULTS: FH binding to D39 and not to TRE108 (PspC-) cells was demonstrated by flow cytometry . Pneumococcal isolates of 14 different strains varied in their ability to bind FH . The binding region of FH within PspC to the first 225 amino acids of the alpha-helical domain was localized . The corresponding binding site for PspC is located within the SCR 6-10 region of FH . Haemolysis of rabbit red blood cells was inhibited by FH even in the presence of PspC . INTERPRETATION & CONCLUSION: FH binding is specific to PspC on the pneumococcal cell surface . The binding region on PspC mapped to the non-conserved N-terminal region of the alpha-helical domain . The binding site on FH to PspC is different from the active site that functions in degradation of C3b . A haemolysis assay provided evidence that the functional activity of FH was maintained upon binding to PspC . Thus, binding of FH to PspC might be an important mechanism by which S . pneumoniae resist complement activation and opsonophagocytosis.

Indian J Med Res, 2004 May, 119 Suppl, 61 - 5
Characterization of the interaction of the pneumococcal surface protein SpsA with the human polymeric immunoglobulin receptor (hpIgR); Elm C et al.; BACKGROUND & OBJECTIVES: The polymeric immunoglobulin receptor (pIgR) is produced by mucosal epithelial cells and plays a crucial role in mucosal immunity . At the basolateral surface of mucosal cells, the pIgR binds predominantly polymeric immunoglobulins, such as dimeric IgA and polymeric IgA (pIgA) and mediates their transport across the polarized cells . This results in apical release of secretory component (SC), either free or bound covalently to IgA, forming secretory IgA (SIgA) . The choline-binding protein (Cbp) SpsA, also called PspC and CbpA, has been shown to interact with the pIgR . A hexapeptide motif in SpsA was identified as the minimal binding motif required for binding specifically to pIgR and SC . The present study was carried out to show that the hexapeptide motif in SpsA is crucial for the interaction of pneumococci and pIgR-expressing cells . METHODS: Streptococcus pneumoniae were cultured to mid-log phase . Calu-3 cells and MDCK epithelial cells, stably transfected with the hpIgR cDNA in pCB6 were used in in vitro infection experiments . Pneumococcal adherence to and invasion of epithelial cells were assayed . RESULTS: By the use of the N-terminal domain of SpsA and SpsA(201), which exhibits a single amino acid substitution in the pIgR-binding motif, in vitro assays indicated the association of the identified hexapeptide motif, located between amino acid 198 and 203 in SpsA, with pneumococcal adherence to and invasion of hpIgR-expressing cells . INTERPRETATION & CONCLUSION: The present findings demonstrated not only the crucial role of the hexapeptide of SpsA, not only for the SpsA-pIgR interaction, but also for adherence and invasion of hpIgR-expressing cells.

Indian J Med Res, 2004 May, 119 Suppl, 37 - 43
Identification & characterisation of the two novel streptococcal pyrogenic exotoxins SPE-L & SPE-M; Thomas P et al.; BACKGROUND & OBJECTIVES: The streptococcal pyrogenic exotoxins (SPEs) are produced by Streptococcus pyogenes and belong to the family of bacterial superantigens, a group of highly mitogenic proteins . The aim of this study was to search unfinished streptococcal genomes for novel superantigens, to generate recombinant proteins from potential open reading frames (ORFs) and to analyse them for superantigen activity . METHODS: The microbial genome database was searched using a TBLASTN search programme . Genotyping of S . equi and S . pyogenes isolates was done using the specific primer pairs . The spe-l and spe-m genes were amplified by PCR . RESULTS: Two novel streptococcal superantigen genes (sepe-l and sepe-m) were identified from the Streptococcus equi genomic database at the Sanger Centre . Genotyping of S . pyogenes isolates resulted in the detection of the orthologous genes spe-l and spe-m in a restricted number of S . pyogenes isolates and revealed a link of spe-l to the M89 serotype . Recombinant SPE-L and rSPE-M were highly mitogenic for human peripheral blood lymphocytes with half maximum responses at 1 pg/ml and 10 pg/ml, respectively . The results from competitive binding experiments suggest that both proteins bind MHC class II at the beta-chain, but not at the alpha-chain . The most common target for both toxins were human Vbetal.1 expressing T cells . Seroconversion against SPE-L and SPE-M was observed in healthy blood donors . INTERPRETATION & CONCLUSION: The two novel ORFs identified in both, S . equi and S . pyogenes, code for proteins that show typical superantigen features . The seroconversion seen in some blood donors suggest that the proteins are indeed produced by the bacteria . Interestingly, the spe-l gene is highly associated with S . pyogenes M89, which is linked to acute rheumatic fever in New Zealand.

Indian J Med Res, 2004 May, 119 Suppl, 33 - 6
Studies of recombinant streptococcal pyrogenic exotoxin B/cysteine protease (rSPE B/SCP) in the skin of guinea pigs & the release of histamine from cultured mast cells & basophilic leukocytes; Ohkuni H et al.; BACKGROUND & OBJECTIVES: Streptococcal pyrogenic exotoxin B/streptococcal cysteine protease (SPE B/SCP) is considered to be one of the virulence factors of Streptococcus pyogenes (S . pyogenes) which causes serious diseases such as severe invasive infections and streptococcal toxic shock syndrome (STSS) . There are no reports on the histamine releasing activity of SPE B/SCP from mast cells, although several biological activities have been studied . It is not clear whether SPE B/SCP have the superantigenic activity . We studied whether SPE B/SCP plays as a pathogenic factor in streptococcal infections and STSS through a histamine releasing activity . METHODS: Human mast cells and basophils were generated from CD34 positive cells isolated from cord blood and cultured in the presence of rIL-6, stem cell factor and/or rIL-3 . The capacity of increasing capillary permeability of recombinant SPE B/SCP (rSPE B/SCP) was studied by using the skin of guinea pigs . Mitogenic activity to human T-cells of rSPE B/SCP was studied by incorporation of (3)Hthymidine . The levels of histamine in the plasma of patients with STSS and controls were measured by ELISA kit . RESULTS: rSPE B/SCP induced increased capillary permeability in the skin of guinea pigs, but both SPE A and SPE C did not exhibit such activity . Histamine was released from cultured human mast cells stimulated with rSPE B/SCP . The rSPE B/SCP did not exhibit mitogenic activity to human T-cells . Three of the 7 patients with STSS showed higher levels of plasma histamine than those of normal subjects . INTERPRETATION & CONCLUSION: The results suggested that increased capillary permeability and histamine release from mast cells induced by rSPE B/SCP might be involved in STSS and/or streptococcal infection of skin and mucous membrane.

Indian J Med Res, 2004 May, 119 Suppl, 29 - 32
Characterization of plasmin(ogen) binding to Streptococcus pneumoniae; Bergmann S et al.; BACKGROUND & OBJECTIVES: The proteolytic activity of plasmin promotes migration of pathogenic bacteria through the human extracellular matrix . The human pathogen Streptococcus pneumoniae binds both human plasminogen and plasmin via the surface displayed alpha-enolase designated Eno . Electron microscopic studies verified the surface exposition of the glycolytic enzyme alpha-enolase and moreover, its ability to reassociate to the cell surface . Carboxyterminal lysine residues of recently described eukaryotic and prokaryotic plasminogen-binding proteins such as SEN of S . pyogenes are involved in interaction with lysine binding sites of kringle domains of plasminogen . In this study, the role of carboxy terminal lysyl residue of eno in plasminogen binding is further analysed . METHODS: Site-directed mutagenesis of eno gene was done using DNA primers with Hind III-restriction enzyme sites for cloning . Purified Eno fusion proteins were separated by SDS-PAGE and human plasminogen binding assay was performed . Radioiodinated ligand binding was done by competitive inhibition assay . RESULTS: Binding assays performed under reduced conditions indicated also a role of the C-terminal lysyl residues of Eno for plasmin(ogen) binding . Binding of pneumococci to radioiodinated plasminogen was competitively inhibited in the presence of plasminogen, kringle 1-3 (LBS 1) and the lysineanalogon epsilon-amino caproic acid indicating the crucial role of lysine-binding sites of plasminogen . However, binding analysis of plasminogen and LBS 1 to wild type Eno and carboxy terminal modified Eno proteins did not reveal any difference in plasminogen-binding activity under native conditions . INTERPRETATION & CONCLUSION: The present results suggested the presence of a further plasminogenbinding motif in Eno . This hypothesis was confirmed by plasminogen-binding activity of reassociated C-terminal modified enolase to the pneumococcal surface and indicated, therefore, the presence of a further binding motif in Eno for plasminogen binding.

Indian J Med Res, 2004 May, 119 Suppl, 13 - 6
Prevention of streptococcal pharyngitis by anti-Streptococcus pyogenes bacteriocin-like inhibitory substances (BLIS) produced by Streptococcus salivarius; Tagg JR; BACKGROUND & OBJECTIVES: Streptococcus salivarius is a numerically prominent member of the human oral microbiota that produces a variety of bacteriocin-like inhibitory substances (BLIS) having in vitro inhibitory activity against S . pyogenes . Our previous studies of S . salivarius isolates from children using a deferred antagonism BLIS production (P)-typing scheme showed that the 9 per cent of children having large populations of P-type 677 S . salivarius experienced fewer S . pyogenes acquisitions than either the 11 per cent of children having predominant P-type 226 populations or the 60 per cent of children with largely non-inhibitory (P-type 000) S . salivarius . Amongst the other BLIS P-types detected were a number of strongly-inhibitory (P-type 777) S . salivarius . In the present study the inhibitory agents produced by prototype strains of P-types 226, 677 and 777 S . salivarius are compared . METHODS: The prototype BLIS-producing S . salivarius strains SN, 20P3, and K12 were isolated from tongue swabbings . BLIS P-typing was done using standard procedures . The BLIS molecules were purified and characterized . RESULTS: S . salivarius SN (P-type 226) produces a heat-labile muramidase . S . salivarius 20P3 (P-type 677) produces the 2315 Da lantibiotic salivaricin A and S . salivarius K12 (P-type 777) produces two lantibiotics; salivaricin A2 (2368 Da) and salivaricin B (2733 Da) . INTERPRETATION & CONCLUSION: The P-type 777 S . salivarius strain produced salivaricin A2 and salivaricin B . The combined production of two anti-S . pyogenes BLIS activities by this strain indicates that it could be adopted as a colonizing strain in bacterial interference trials.

Indian J Med Res, 2004 May, 119 Suppl, 7 - 12
Short-chain oligosaccharide protein conjugates as experimental pneumococcal vaccines; Jansen WT et al.; Streptococcus pneumoniae remains a major cause of acute respiratory infections worldwide and is responsible for approximately 1 million childhood deaths each year . Despite the widespread use of antibiotics, the mortality and morbidity of pneumococcal disease remains high . Therefore, effective vaccines to prevent pneumococcal disease are needed . Bacterial vaccine development in general follows a similar track starting from large, rather crude vaccines (whole live, attenuated pathogen) towards smaller, better defined subunit vaccines . Ultimately, this track leads to the development and evaluation of minimal, highly defined subunit vaccines, based on a collection of single protective epitopes . This mini-review deals with capsular saccharide based vaccines . After a short overview of the development of pneumococcal vaccines from the 23 - valent polysaccharide vaccines to polysaccharide-protein conjugate vaccines, it focuses on the vaccine potential of synthetic oligosaccharides, conjugated to carrier proteins.

J Bacteriol, 2004 Jul, 186(14), 4740 - 7
Distribution, genetic diversity, and variable expression of the gene encoding hyaluronate lyase within the Streptococcus suis population; King SJ et al.; Although Streptococcus suis is an economically important pathogen of pigs and an occasional cause of zoonotic infections of humans knowledge of crucial virulence factors, and as a consequence targets for therapeutic or prophylactic intervention, remains limited . Here we describe a detailed study of the distribution, diversity, and in vitro expression of hyaluronate lyase, a protein implicated as a virulence factor of many mucosal pathogens . The gene encoding hyaluronate lyase, hyl, was present in all 309 bona fide S . suis isolates examined representing diverse serotypes, geographic sources, and clinical backgrounds . Examination of the genetic diversity of hyl by RFLP and sequence analysis indicated a pattern of diversity shared by many gram-positive surface proteins with a variable 5' region encoding the most distal cell surface-exposed regions of the protein and a much more conserved 3' region encoding domains more closely associated with the bacterial cell . Variation occurs by several mechanisms, including the accumulation of point mutations and deletion and insertion events, and there is clear evidence that genetic recombination has contributed to molecular variation in this gene . Despite the ubiquitous presence of hyl, the corresponding enzyme activity was detected in fewer than 30% of the 309 isolates . In several cases this lack of activity correlates with the presence of mutations (either sequence duplications or point mutations) within hyl that result in a truncated polypeptide . There is a striking absence of hyaluronate lyase activity in a large majority of isolates from classic S . suis invasive disease, indicating that this protein is probably not a crucial virulence factor, although activity is present in significantly higher numbers of isolates associated with pneumonia .

J Bacteriol, 2004 Jul, 186(14), 4638 - 44
Differences in membrane fluidity and fatty acid composition between phenotypic variants of Streptococcus pneumoniae; Aricha B et al.; Phase variation in the colonial opacity of Streptococcus pneumoniae has been implicated as a factor in the pathogenesis of pneumococcal disease . This study examined the relationship between membrane characteristics and colony morphology in a few selected opaque-transparent couples of S . pneumoniae strains carrying different capsular types . Membrane fluidity was determined on the basis of intermolecular excimerization of pyrene and fluorescence polarization of 1,6-diphenyl 1,3,5-hexatriene (DPH) . A significant decrease, 16 to 26% (P < or = 0.05), in the excimerization rate constant of the opaque variants compared with that of the transparent variants was observed, indicating higher microviscosity of the membrane of bacterial cells in the opaque variants . Liposomes prepared from phospholipids of the opaque phenotype showed an even greater decrease, 27 to 38% (P < or = 0.05), in the pyrene excimerization rate constant compared with that of liposomes prepared from phospholipids of bacteria with the transparent phenotype . These findings agree with the results obtained with DPH fluorescence anisotropy, which showed a 9 to 21% increase (P < or = 0.001) in the opaque variants compared with the transparent variants . Membrane fatty acid composition, determined by gas chromatography, revealed that the two variants carry the same types of fatty acids but in different proportions . The trend of modification points to the presence of a lower degree of unsaturated fatty acids in the opaque variants compared with their transparent counterparts . The data presented here show a distinct correlation between phase variation and membrane fluidity in S . pneumoniae . The changes in membrane fluidity most probably stem from the observed differences in fatty acid composition .

BMC Infect Dis . 2004 Jun 30;4(1):21.
Serotype-specific mortality from invasive Streptococcus pneumoniae disease revisited; Martens P et al.; BACKGROUND: Invasive infection with Streptococcus pneumoniae (pneumococci) causes significant morbidity and mortality . Case series and experimental data have shown that the capsular serotype is involved in the pathogenesis and a determinant of disease outcome . METHODS: Retrospective review of 464 cases of invasive disease among adults diagnosed between 1990 and 2001 . Multivariate Cox proportional hazard analysis . RESULTS: After adjustment for other markers of disease severity, we found that infection with serotype 3 was associated with an increased relative risk (RR) of death of 2.54 (95% confidence interval (CI): 1.22-5.27), whereas infection with serotype 1 was associated with a decreased risk of death (RR 0.23 (95% CI, 0.06-0.97)) . Additionally, older age, relative leucopenia and relative hypothermia were independent predictors of mortality . CONCLUSION: Our study shows that capsular serotypes independently influenced the outcome from invasive pneumococcal disease . The limitations of the current polysaccharide pneumococcal vaccine warrant the development of alternative vaccines . We suggest that the virulence of pneumococcal serotypes should be considered in the design of novel vaccines.

J Vet Med B Infect Dis Vet Public Health, 2004 May, 51(4), 180 - 4
Polymerase chain reaction mediated identification of Streptococcus uberis and Streptococcus parauberis using species-specific sequences of the genes encoding superoxide dismutase A and chaperonin 60*; Alber J et al.; Streptococcus uberis, a well-known bacterial pathogen associated with bovine mastitis, appears to be biochemically and serologically almost indistinguishable from the closely related species Streptococcus parauberis . In the present study, species-specific oligonucleotide primers were designed using internal parts of the genes sodA, encoding superoxide dismutase A, and cpn60 encoding chaperonin 60 of S . uberis and S . parauberis, respectively . The two oligonucleotide primer pairs allowed a rapid and reliable PCR-mediated identification and differentiation of both species . These studies, performed with S . uberis and S . parauberis reference cultures and clinical isolates from routine diagnostics, revealed that the occurrence of S . parauberis as causative agent of bovine mastitis appears to be rare . In addition the sodA and cpn60 sequence data confirmed that both species could taxonomically be classified to the pyogenic group of genus Streptococcus.

Biochem J, 2004 Oct 1, 383(Pt 1), 159 - 64
Binding of salivary agglutinin to IgA; Ligtenberg AJ et al.; SAG (salivary agglutinin), which is identical to gp-340 (glycoprotein-340) from the lung, is encoded by DMBT1 (deleted in malignant brain tumours 1) . It is a member of the SRCR (scavenger receptor cysteine-rich) superfamily and contains 14 SRCR domains, 13 of which are highly similar . SAG in saliva is partially complexed with IgA, which may be necessary for bacterial binding . The goal of the present study was to characterize the binding of purified SAG to IgA . SAG binds to a variety of proteins, including serum and secretory IgA, alkaline phosphatase-conjugated IgGs originating from rabbit, goat, swine and mouse, and lactoferrin and albumin . Binding of IgA to SAG is calcium dependent and is inhibited by 0.5 M KCl, suggesting that electrostatic interactions are involved . Binding of IgA was destroyed after reduction of SAG, suggesting that the protein moiety is involved in binding . To pinpoint further the binding domain for IgA on SAG, a number of consensus-based peptides of the SRCR domains and SRCR interspersed domains were designed and synthesized . ELISA binding studies with IgA indicated that only one of the peptides tested, comprising amino acids 18-33 (QGRVEVLYRGSWGTVC) of the 109-amino-acid SRCR domain, exhibited binding to IgA . This domain is identical to the domain of SAG that is involved in binding to bacteria . Despite this similar binding site, IgA did not inhibit binding of Streptococcus mutans to SAG or peptide . These results show that the binding of IgA to SAG is specifically mediated by a peptide sequence on the SRCR domains.

Ann Clin Lab Sci, 2004 Spring, 34(2), 131 - 7
Review: Drug-induced neutropenia--pathophysiology, clinical features, and management; Bhatt V et al.; Drug therapy plays a significant role in causing neutropenia . The neutropenia may be immune mediated or due to direct inhibition of the bone marrow precursors . Recently, due to wide use of chemotherapy, febrile neutropenia has become a common and devastating problem . Neutropenia predisposes to many bacterial and fungal infections with organisms including gram negative bacilli such as E . coli, Klebsiella, and Pseudomonas; gram positive organisms such as Staphylococcus, Streptococcus viridans, and Enterococcus species; and fungi, like Candida and Aspergillus . In addition to the customary supportive care for neutropenic patients, therapy with recombinant human granulocyte colony-stimulating factor (rG-CSF) (filgrastim) has been shown to be beneficial . Filgrastim was a significant advance in the management of drug induced neutropenia in the past decade, but therapy with pegfilgrastim (a pegylated, long-acting form of filgrastim) ushers in the current decade . Pegfilgrastim (Neulasta) is administered as a single s.c . injection once per chemotherapy cycle . This results in fewer injections, fewer patient visits to the physician's office, and better patient compliance with therapy.

Aust Fam Physician, 2004 May, 33(5), 297 - 301
Lower respiratory tract infections and community acquired pneumonia in adults; Stocks N et al.; BACKGROUND: Lower respiratory tract infections--acute bronchitis and community acquired pneumonia (CAP)--are important causes of morbidity in Australia . Acute bronchitis is often treated with antibiotics, although the cause is usually viral . Community acquired pneumonia may be fatal, particularly in the elderly, therefore appropriate assessment and management is essential . OBJECTIVE: This article describes the aetiology, clinical assessment, investigations and management of acute bronchitis and CAP in the community . DISCUSSION: Clinical assessment is important for acute bronchitis and CAP, with investigations such as C reactive protein, serology, and chest X-ray informing diagnosis and management of the latter . Causative organisms are usually not identified, but are presumed to be viral for acute bronchitis, and Streptococcus pneumoniae for CAP; although 'atypicals' are also important . Antibiotics should generally not be prescribed for acute bronchitis, however, there is some evidence they may provide limited benefits in patients who have chest signs, are very unwell, are older, have comorbidities, or smoke . In patients with CAP, treated outside of hospital, the combination of amoxycillin and doxycycline/roxithromycin is the treatment of choice.

Ann Pharmacother, 2004 Sep, 38(9 Suppl), S8 - S13 Epub 2004 Jun 29.
Increased bacterial resistance: PROTEKT US--an update; Rybak MJ; Community-acquired respiratory tract infections (CARTIs), including community-acquired pneumonia, acute exacerbations of chronic bronchitis, and acute bacterial sinusitis, remain substantial sources of morbidity and mortality in the US . The increasing prevalence of antimicrobial-resistant strains among the common respiratory pathogens, particularly Streptococcus pneumoniae, has complicated the choice of antimicrobials and threatens the efficacy of treatment for CARTIs . This highlights the need both for ongoing surveillance efforts to track current resistance patterns and for the development of new antimicrobial agents effective against resistant organisms . To provide current and ongoing antimicrobial resistance surveillance data, including comparisons of telithromycin (the first ketolide to undergo clinical development and be approved by the Food and Drug Administration) with other agents, a longitudinal global surveillance study--Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin (PROTEKT)--was launched in 1999 . A US component of this study was initiated in 2000, and results from the first respiratory infection tracking season (2000-2001) are discussed in this article . Providing resistance data on the national, regional, and local levels, PROTEKT US has thus far confirmed the increasing prevalence of resistant S . pneumoniae isolates to beta-lactams and macrolides, as well as the emergence of fluoroquinolone resistance in the US . Importantly, telithromycin was highly active against S . pneumoniae, including resistant isolates.

Int J Antimicrob Agents, 2004 Jul, 24(1), 79 - 82
In vitro combined effect of co-amoxiclav concentrations achievable in serum after a 2000/125 mg oral dose, and polymorphonuclear neutrophils against strains of Streptococcus pneumoniae exhibiting decreased susceptibility to amoxicillin; Amores R et al.; The in vitro effect that the presence of components of non-specific immunity (serum plus polymorphonuclear neutrophils) has on the bactericidal activity of co-amoxiclav was explored against Streptococcus pneumoniae strains exhibiting an amoxicillin MIC > or =4 mg/L . Eight penicillin-resistant clinical isolates non-susceptible to co-amoxiclav with MICs of 4 (two strains), 8 (four strains) and 16 mg/L (two strains) were used . Values of MBC were identical to MIC values in all cases . Time-kill curves were performed with co-amoxiclav concentrations achievable in serum after a single oral dose administration of the new 2000/125 mg sustained-release formulation . Results were expressed as percentage of reduction of initial inocula after 3 h incubation . Control curves showed growth with no reduction of initial inocula . Against strains with MIC of 4 and 8 mg/L, the results obtained with the antibiotic alone or with the presence of factors of non-specific immunity were similar, with a weak combined effect due to the intrinsic activity of co-amoxiclav (reductions of initial inocula ranging from 70 to 99.16%) . Against strains with MIC of 16 mg/L, the addition of PMN in the presence of serum increased the reduction of bacterial load provided by the aminopenicillin, even at sub-inhibitory concentrations (25.8% versus 51.1% at 0.5 x MIC concentration--8/0.5 mg/L) . This combined activity against strains with an amoxicillin MIC of 16 mg/L which decreased the bacterial load may be important in preventing bacterial proliferation within the host and the transmission of resistant clones to others.

Int J Antimicrob Agents, 2004 Jul, 24(1), 43 - 7
Molecular characterisation of macrolide resistance mechanisms of Streptococcus pneumoniae and Streptococcus pyogenes isolated in Germany, 2002-2003; Reinert RR et al.; In the present study, a real-time PCR protocol was developed for the detection of macrolide resistance determinants and was validated in a nationwide study in Germany covering a total of 236 Streptococcus pyogenes and 241 Streptococcus pneumoniae strains collected from children < or = 16 years of age with community-acquired infections . Macrolide resistance was observed in 19.9% of pneumococcal strains and 14% of S . pyogenes isolates . Of the erythromycin A-resistant S . pyogenes strains, 93.9% showed the efflux type mef(A); 62.5% of the S . pneumoniae strains were mef(A)- and 37.5% erm(B)-positive . The correlation of the results of real-time PCR assay genotyping in the present study compared with those of conventional PCR genotyping and resistance phenotyping was 100% . Macrolide resistance is of growing concern in Germany . This highly sensitive and specific PCR assay to detect macrolide resistance has the potential to provide sufficiently rapid results to improve antibiotic treatment of streptococcal infections.

Int J Antimicrob Agents, 2004 Jul, 24(1), 39 - 42
Comparative in vitro activity of antiribosomal agents on penicillin-susceptible and -resistant Streptococcus pneumoniae in relation to their resistance genotypes; Sener B et al.; A collection of 326 strains of Streptococcus pneumoniae isolated from blood, cerebrospinal fluid, bronchoalveolar lavage, transtracheal aspirate or sputum from January 1996-June 2002 were included in this study . The activity of clarithromycin, clindamycin, telithromycin, linezolid and quinupristin/dalfopristin against penicillin G and erythromycin A susceptible and resistant pneumococci were determined; the erythromycin A resistance phenotypes and genotypes were identified and susceptibilities of these agents were assessed according to the resistance genotypes . MICs were determined for all strains of pneumococci using an agar dilution method . MLS(B) resistance phenotypes were determined by the double disk (erythromycin A and clindamycin) diffusion method . Genetic determinants for macrolide resistance were identified by PCR using primers specific for erm(B) and mef(A) . Erythromycin A resistance was detected in 13.8% of the strains . MLS(B) resistance phenotype was observed in 82% of these (60% being cMLS(B) and 40% being iMLS(B)), and M type resistance in about 18% . All the MLS(B) phenotype strains except four, revealed the presence of erm(B) gene and all except one M phenotype strains revealed the mef(A) gene . Of the erythromycin A resistant pneumococci about 49% were also resistant to clindamycin . No strains were resistant to telithromycin, quinupristin/dalfopristin and linezolid . Telithromycin had the lowest MIC values for both erythromycin A resistant and susceptible strains of all the antiribosomal agents tested . The most prevalent mechanism of macrolide resistance was mediated by the erm(B) gene leading to the expression of MLS(B) phenotype . Telithromycin was the most active antiribosomal agent, regardless of the macrolide resistance genotype of the pneumococci tested.

Int J Antimicrob Agents, 2004 Jul, 24(1), 1 - 17
Clarithromycin in 2003: sustained efficacy and safety in an era of rising antibiotic resistance; Anzueto A et al.; Data from surveillance studies show increasing prevalence of respiratory pathogens resistant to commonly used antibiotics . Thus, a Medline search was conducted to identify studies of clarithromycin, especially those addressing macrolide resistance . Changing trends of in vitro susceptibility have not affected clinical efficacy with clarithromycin . Over the last 12 years, clarithromycin study results have shown consistent rates of clinical cure and bacteriological eradication, which are similar to those observed with comparator agents . The incidence of clarithromycin treatment failure in patients infected with Streptococcus pneumoniae is substantially less than that predicted by macrolide resistance rates from surveillance programmes . In summary, despite widespread use since its introduction, clarithromycin remains active both in vitro and in vivo against clinically relevant respiratory tract pathogens.

Transpl Infect Dis, 2004 Mar, 6(1), 41 - 5
Spondylodiscitis after bilateral sequential lung transplantation in a patient with cystic fibrosis; Quattrucci S et al.; We report the case of a 24-year-old woman with cystic fibrosis in whom spondylodiscitis developed after bilateral sequential transplantation . The diagnostic work-up included magnetic resonance imaging, computed tomography-guided disk biopsy, histological examination, and cultures of disk specimens . The infective organism was D group Streptococcus and the patient was successfully treated with intravenous piperacillin followed by oral ampicillin . To our knowledge, this is the first reported case of spondylodiscitis after lung transplantation.

Vet Microbiol, 2004 Jul 14, 101(3), 215 - 21
In vitro growth inhibition of major mastitis pathogens by Staphylococcus chromogenes originating from teat apices of dairy heifers; De Vliegher S et al.; Earlier field observations suggest that teat apex colonization by Staphylococcus chromogenes pre-partum in dairy heifers protects udder quarters against elevated somatic cell counts early post-partum . To explain these findings, the in vitro inhibitory capability of S . chromogenes from teat apices of heifers towards some major mastitis pathogens was tested using a modified cross-streaking method . Two out of 10 S . chromogenes isolates, both originating from two different teats from the same heifer, consistently inhibited growth of all Staphylococcus aureus, Streptococcus dysgalactiae, and Streptococcus uberis strains, but none of the Escherichia coli strains . The present study, therefore, supports the protective effect of teat apex colonization by S . chromogenes by in vitro production of inhibitory substances.

Biochem Biophys Res Commun, 2004 Jul 23, 320(2), 347 - 53
Identification and characterization of bacterial-binding property in the type III repeat domain of fibronectin; Ito HO et al.; To characterize fibronectin binding with Granulicatella adiacens, a causative agent of infective endocarditis, monoclonal antibodies were generated against human fibronectin and selected for their capacity to inhibit the fibronectin binding of the organism . Thermolysin and lysyl-endopeptidase digests of fibronectin were characterized by Western blot . The epitope of inhibitory monoclonal antibody was found in the central portion of fibronectin known as the cell-binding domain, and not in the N-terminal portion known to be the binding region of most microbial species, e.g., Staphylococcus aureus and Streptococcus pyogenes . While these two species could bind to both the N-terminal and central portion, Escherichia coli and G . adiacens bind only to the latter . Excess amounts of free fibronectin in the solution inhibited the bacterial adherence to the N-terminal fibronectin fragment, but not to the central region, thereby suggesting the central region plays a significant role for in vivo bacterial colonization in the presence of high concentrations of soluble fibronectin.

Jpn J Antibiot, 2004 Apr, 57(2), 172 - 86
{Epidemiology of Streptococcus pneumoniae isolates in Gifu Prefecture}; Mikamo H et al.; We analyzed Streptococcus pneumoniae isolates confirmed by direct PCR in Gifu prefecture between May 2002 and August 2002 . We analyzed isolates of 254 strains from 6 hospitals to determine antibiotic susceptibility, genotype of penicillin-binding protein (PBP) genes and macrolide resistant genes, and the serotypes distribution of isolates from Matsubara Otorhinolaryngology Clinic . Isolates in which abnormal PBP genes of pbp1a, pbp2x, and pbp2b were identified by PCR were classified based on PCR results as follows; (i) penicillin-susceptible (PSSP) with 3 normal PBP genes, (ii) penicillin-intermediate (PISP) with an abnormal pbp2x, (iii) PISP with an abnormal php2b, (iv) PISP with abnormal pbp2x and pbp2b, (v) PISP with abnormal pbpla and pbp2x, (vi) penicillin-resistant (PRSP) with 3 abnormal PBP genes . The overall incidence of PRSP, PISP and PSSP was 121 (49%), 109 (42%) and 24 (9%), respectively, and there was a significant difference among some hospitals (p<0.05) . However, there was no significant difference among the hospitals for the incidence of abnormal macrolide-resistant genes (mefA, ermB) . Panipenem showed an excellent antimicrobial activity for injectable carbapenems against PRSP, following biapenem, imipenem, and meropenem . Cefditoren (CDTR) showed an excellent antimicrobial activity for oral cephalosporins against PRSP, following cefteram and cefcapene . Interestingly, there were 2 and 3 strains on MIC of CDTR for 8 and 4 microg/mL, respectively . The prevalent pneumococcal serotypes of isolates in Matsubara Clinic were 6 (17/55), following by 40 (8/55), 9 (6/5) and 15 (5/55) . The endemic strains were observed in this study using pulsed field gel electrophoresis . These findings suggest the needs to continue the surveillance of bacterial resistance not only in the nationwide but also in the distict.

J Dent Res, 2004 Jul, 83(7), 534 - 9
Streptococcus mutans strains harboring collagen-binding adhesin; Sato Y et al.; A previously unidentified 120-kDa protein was detected in Streptococcus mutans strain Z1 and was involved in the cold-agglutination of the strain . We have identified the gene, designated cnm, as being involved in the agglutination of strain Z1 following random mutagenesis . The amino acid sequence of the deduced Cnm protein exhibited high similarity to those of collagen-binding adhesins from staphylococci and other organisms . To confirm whether the protein is involved in collagen-binding, we cloned a cnm gene fragment, overexpressed it in E.coli, and prepared crude extracts . The extracts containing recombinant protein exhibited binding to immobilized collagen and laminin but not to fibronectin . Compared with the parental strain Z1, the cold-agglutination-negative mutant 05A02 exhibited reduced binding to collagen and laminin but retained that to fibronectin . This gene was detected in some strains of S . mutans . Therefore, the cnm gene encoded a new strain-specific member of the collagen-binding adhesin family.

J Chemother, 2004 Apr, 16(2), 122 - 7
Increased microbicidal activity of green tea (Camellia sinensis) in combination with butylated hydroxyanisole; Simonetti G et al.; We have demonstrated that green tea (Camellia sinensis) shows increased antimicrobial activity against bacteria and fungi when used in combination with butylated hydroxyanisole (BHA) . Glycolic extract taken from green tea showed only limited activity against Streptococcus mutans and no activity against Candida albicans and certain strains of Escherichia coli . BHA, at non inhibitory concentrations, increased the microbicidal activity of green tea against 10(10) S . mutans (p<0.01), non-susceptible E . coli (p<0.01) and C . albicans (p<0.01) . Green tea in combination with BHA reduced the hydrophobicity of S . mutans (p<0.01) and greatly inhibited (p<0.001) the formation of hyphae in C . albicans . The increased antimicrobial activity of green tea is related to an impairment of the barrier function in microorganisms and a depletion of thiol groups . The increased activity of green tea as an oral antimicrobial product is discussed.

Microbiol Immunol, 2004, 48(6), 449 - 56
Application of in vitro mutagenesis to identify the gene responsible for cold agglutination phenotype of Streptococcus mutans; Sato Y et al.; A previously unidentified protein with an apparent molecular mass of 120 kDa was detected in some Streptococcus mutans strains including the natural isolate strain Z1 . This protein was likely involved in the cold-agglutination of the strain, since a correlation between this phenotype and expression of the 120 kDa protein was found . We have applied random mutagenesis by in vitro transposition with the Himar1 minitransposon and isolated three cold-agglutination-negative mutants of this strain from approximately 2,000 mutants screened . A 2.5 kb chromosomal fragment flanking the minitransposon in one of the three mutants was amplified by PCR-based chromosome walking and the minitransposon insertion in the other two mutants occurred also within the same region . Nucleotide sequencing of the region revealed a 1617 nt open reading frame specifying a putative protein of 538 amino acid residues with a calculated molecular weight of 57,192 . The deduced eight amino acid sequence following a putative signal sequence completely coincided with the N-terminal octapeptide sequence of the 120 kDa protein determined by the Edman degradation . Therefore, the 1617 nt gene unexpectedly encoded the 120 kDa protein from S . mutans . Interestingly, this gene encoded a collagen adhesin homologue . In vitro mutagenesis using the Himar1 minitransposon was successfully applied to S . mutans.

J Biol Chem, 2004 Aug 27, 279(35), 36426 - 32 Epub 2004 Jun 23.
Nucleotide-binding oligomerization domain proteins are innate immune receptors for internalized Streptococcus pneumoniae; Opitz B et al.; Streptococcus pneumoniae, the major cause of community-acquired pneumonia and bacterial meningitis, has been shown to transiently invade epithelial and endothelial cells . Innate immune receptors including Toll-like receptors recognize various pathogens, such as S . pneumoniae, by identifying conserved pathogen-associated molecular patterns . Recently, two members of a novel class of pattern recognition receptors, the cytosolic proteins nucleotide-binding oligomerization domain 1 (Nod1)/CARD4 and Nod2/CARD15, have been found to detect cell wall peptidoglycans . Here we tested the hypothesis that Nod proteins are involved in the intracellular recognition of pneumococci . Data indicate that pneumococci invade HEK293 cells . Genetic complementation studies in these cells demonstrate that NF-kappaB activation induced by S . pneumoniae depends on Nod2 . Moreover, intracellular transfection of inactivated pneumococci yielded similar effects, confirming the Nod2 dependence of NF-kappaB activation by pneumococci in HEK293 cells . By dominant negative overexpression and small interfering RNA experiments, we show for the first time that interleukin-1 receptor-associated kinase participates in Nod2-dependent NF-kappaB activation . Additionally, dominant negative interleukin-1 receptor-associated kinase 2, tumor necrosis factor receptor-associated factor 6, NF-kappaB-inducing kinase, transforming growth factor-beta-activated kinase-binding protein 2, and transforming growth factor-beta-activated kinase 1 also inhibited Nod2-dependent NF-kappaB activation . We finally demonstrate that in C57BL/6 mouse lung tissue in vivo as well as in the bronchial epithelial cell line BEAS-2B, Nod1 and Nod2 mRNA expressions were up-regulated after pneumococcal infection . Data presented suggest that Nod proteins contribute to innate immune recognition of S . pneumoniae . Furthermore, Rip-2 and members of the Toll-like receptor-signaling cascade are involved in the Nod2-dependent activation of NF-kappaB induced by pneumococci.

Glycobiology, 2004 Oct, 14(10), 931 - 8 Epub 2004 Jun 23.
Hyaluronan release from Streptococcus pyogenes: export by an ABC transporter; Ouskova G et al.; Capsular hyaluronan of Streptococcus pyogenes is synthesized at the protoplast membrane . It is widely assumed that hyaluronan is exported by the synthase itself and that no additional protein is required for transfer through plasma membranes . However, we produced an insertional mutation that reduced the mucoid phenotype, hyaluronan production, and capsule formation . Nucleotide sequence analysis of the insertion site identified a gene coding for a protein with an ATP-binding cassette (ABC) that belonged to an ABC transporter system and was located next to the hyaluronan synthesis genes . The mucoid phenotype was reconstituted by complementation with DNA encoding the ABC transporter system . These results indicated that an ABC transporter was required for efficient capsule production.

Antimicrob Agents Chemother, 2004 Jul, 48(7), 2757 - 9
rpoB mutations in Streptococcus mitis clinical isolates resistant to rifampin; Achour W et al.; Activity of rifampin against 129 Streptococcus mitis isolates obtained from patients with hematologic cancer was investigated . One hundred twenty-five strains were susceptible to rifampin, and 4 were resistant (MIC = 32 to 64 microg/ml) . Resistance to rifampin was related to mutations in the rpoB gene: His(526)Asn in three strains and His(526)Asp in one strain.

Clin Microbiol Infect, 2004 Jul, 10(7), 659 - 62
High-level fluoroquinolone resistance in a clinical Streptoccoccus pyogenes isolate in Germany; Reinert RR et al.; An isolate of Streptococcus pyogenes isolated from a 63-year-old woman with a serious wound infection was found to be highly resistant to fluoroquinolones (levofloxacin MIC > or = 32 mg/L) . DNA amplification and sequencing revealed a serine-81 to phenylalanine substitution in gyrA and three substitutions in parC: serine-79 to phenylalanine, aspartic acid-91 to asparagine, and serine-140 to proline . To our knowledge, this is the first report from a European country of a clinical isolate of S . pyogenes with high-level fluoroquinolone resistance.

Clin Microbiol Infect, 2004 Jul, 10(7), 657 - 9
Antimicrobial susceptibility of invasive isolates of Streptococcus pneumoniae in Ireland; Clarke P et al.; Between January 1999 and June 2002, 646 invasive isolates of Streptococcus pneumoniae were collected in Ireland . MICs of penicillin, ciprofloxacin, cefotaxime, moxifloxacin and linezolid were determined by Etest methodology . Eighty-seven (13.5%) isolates showed intermediate resistance to penicillin, while seven (1.1%) showed high-level resistance . Eighty-seven (13.5%) isolates were resistant to erythromycin, but all isolates were susceptible to cefotaxime, moxifloxacin and linezolid . The prevalence of pneumococcal isolates non-susceptible to penicillin in Ireland is worryingly high, but currently there are alternative agents available to treat invasive infection.

Clin Microbiol Infect, 2004 Jul, 10(7), 652 - 6
Invasive Streptococcus pneumoniae from Portugal: implications for vaccination and antimicrobial therapy; Serrano I et al.; The distribution of pneumococcal serotypes among 465 invasive isolates recovered from 1999 to 2002 in Portugal was analysed by age group . Serotype 14 was either the most prevalent or the second most prevalent in all age groups . Among children aged < 2 years, serotypes 6B and 23F, which are usually associated with children, together with serotypes 19A and 14, accounted for more than half of the isolates . In contrast, in older adults (> or = 60 years), serotypes 3, 14, 1, 8 and 4 were the most prevalent . The potential coverage of the seven-valent conjugate vaccine is 63.2% among infants, and does not change significantly if children aged < 6 years are considered, which is a lower coverage than in other European countries . The potential coverage of the 23-valent polysaccharide vaccine is high in all age groups, particularly among older adults (80.7%) . All isolates were tested for their susceptibility to penicillin, cefuroxime, cefotaxime, vancomycin, erythromycin, clindamycin, levofloxacin, gatifloxacin, moxifloxacin, linezolid, quinupristin-dalfopristin, tetracycline, chloramphenicol and trimethoprim-sulphamethoxazole . Most isolates collected from children aged < 6 years had decreased susceptibility to at least one antibiotic class, whereas isolates from patients aged > or = 6 years were mostly susceptible to all antimicrobial agents tested . Overall, 23% of isolates showed reduced susceptibility to penicillin . Most (98.5%) isolates remained fully susceptible to cefotaxime, and a single isolate was resistant to quinolones.

Clin Microbiol Infect, 2004 Jul, 10(7), 645 - 51
Antimicrobial susceptibility of Streptococcus pneumoniae in Latin America: results from five years of the SENTRY Antimicrobial Surveillance Program; Castanheira M et al.; A total of 1561 pneumococcal isolates were collected in 1997-2001, mainly from patients with community-acquired respiratory tract infections, and susceptibilities were tested by reference broth microdilution against 29 antimicrobial agents . In general, 69.3% of strains were considered susceptible (MIC < or = 0.06 mg/L) to penicillin . Resistance to penicillin (MIC > or = 2 mg/L) and cefotaxime (MIC > or = 4 mg/L) was found in 11.9% and 0.4% of isolates, respectively . The fluoroquinolones gatifloxacin (MIC90, 0.5 mg/L) and levofloxacin (MIC90, 1 mg/L) were active against > 99% of the isolates tested . Among the other non-beta-lactam drugs tested, the rank order of susceptibility was chloramphenicol (95.6%) > clindamycin (94.5%) > azithromycin (88.5%) > clarithromycin (87.5%) >tetracycline (79.5%) > trimethoprim + sulphamethoxazole (60.5%) . The penicillin-non-susceptible isolates presented higher rates of resistance to other antimicrobial agents . The rank order of penicillin resistance rates among the seven participating countries was Mexico (25.0%) > Uruguay (19.2%) > Chile (18.3%) > Colombia = Argentina (9.9%) > Brazil (3.9%) > Venezuela (2.8%) . The regional rate of penicillin resistance did not vary significantly over the years studied (p 0.339) . Screening for the ermB and mefA genes by multiplex rapid cycle PCR on 23 erythromycin-resistant isolates collected during the year 2001 showed that 43.5% and 56.5%, respectively, were positive for ermB and mefA . Overall, the results indicated that antimicrobial susceptibilities of Streptococcus pneumoniae vary significantly among Latin American countries . Regional and local surveillance programmes are necessary to guide empirical therapy of pneumococcal infection in Latin American countries.

Clin Microbiol Infect, 2004 Jul, 10(7), 615 - 23
Evaluation of 5-day therapy with telithromycin, a novel ketolide antibacterial, for the treatment of tonsillopharyngitis; Norrby SR et al.; A pooled analysis of two double-blind, multicentre, Phase III studies compared oral telithromycin 800 mg once-daily for 5 days with penicillin V 500 mg three-times-daily or clarithromycin 250 mg twice-daily for 10 days in the treatment of Streptococcus pyogenes (group A beta-haemolytic streptococcus; GABHS) tonsillopharyngitis . Patients aged > or = 13 years with acute GABHS tonsillopharyngitis were randomised to receive telithromycin (n = 430), penicillin (n = 197) or clarithromycin (n = 231) . Clinical isolates of S . pyogenes (n = 590) obtained from throat swab samples on study entry were tested for their in-vitro susceptibility to telithromycin, clarithromycin and azithromycin . Telithromycin demonstrated in-vitro activity against the clinical isolates of S . pyogenes (MIC50/90 0.03/0.06 mg/L) higher than clarithromycin or azithromycin (MIC50/90 0.06/0.06 mg/L and 0.12/0.25 mg/L, respectively), including erythromycin-resistant strains . At the post-therapy/test of cure (TOC) visit (days 16-23), satisfactory bacteriological outcome was demonstrated for 88.3% (234/265) and 88.6% (225/254) of telithromycin- and comparator-treated patients, respectively (per-protocol population) . Overall, GABHS eradication rates were 88.7% (235/265) for telithromycin and 89.0% (226/254) for comparators . The clinical cure rates at the post-therapy/TOC visit were 93.6% (248/265) and 90.9% (220/242) for telithromycin and pooled comparators, respectively . Telithromycin was generally well-tolerated . Most adverse events considered to be possibly related to study medication were gastrointestinal and of mild intensity . Discontinuations as a result of adverse events were few in both treatment groups . In conclusion, telithromycin 800 mg once-daily for 5 days was as effective as penicillin V or clarithromycin for 10 days in the treatment of GABHS tonsillopharyngitis.

Arch Pathol Lab Med, 2004 Jul, 128(7), 801 - 3
Phlegmonous gastritis associated with Kaposi sarcoma: a case report and review of the literature; Yu QQ et al.; We report a case of phlegmonous gastritis associated with Kaposi sarcoma in a 37-year-old, human immunodeficiency virus (HIV)-positive man who presented with an acute abdomen . Computed tomographic scan revealed free fluid in the abdominal cavity and a thickened gastric wall . A partial gastrectomy was performed . The resected portion of stomach had a hemorrhagic, necrotic thickened wall and showed extensive, acute suppurative inflammation, especially in the submucosa, with focal transmural involvement . Beneath an area of healing ulceration, a focus of Kaposi sarcoma was present . Group A beta-hemolytic streptococcus was grown from peritoneal fluid, and treatment with numerous antibiotics was initiated . After a difficult postoperative course that responded to 8 weeks of antibiotic therapy, the patient was medically stable and discharged from the hospital on antiretroviral therapy for HIV . Phlegmonous gastritis is a rare and rapidly progressive bacterial infection of the gastric wall . Kaposi sarcoma is one of the most common malignancies in HIV-positive patients, and gastric involvement is relatively common in those patients with systemic Kaposi sarcoma . To our knowledge, this is the first reported case of phlegmonous gastritis associated with Kaposi sarcoma, and it represents a rare survival following surgical and antibiotic therapy.

Lett Appl Microbiol, 2004, 38(4), 251 - 6
Bacteriocin-like substance production by Bacillus licheniformis strain P40; Cladera-Olivera F et al.; AIMS: To investigate the production of bacteriocin-like compounds by Bacillus spp . isolated from the Amazon basin . METHODS AND RESULTS: An antimicrobial substance produced by Bacillus licheniformis strain P40 was inhibitory to a broad range of indicator strains, such as Listeria monocytogenes, Bacillus cereus and clinical isolates of Streptococcus spp . The compound was stable at 100 degrees C, but lost its activity when treated at 121 degrees C/103.5 kPa for 15 min . It was resistant to the proteolytic action of trypsin and papain but sensitive to pronase E and was stable within a wide range of pH (3-11) . The substance was bactericidal and bacteriolytic to L . monocytogenes . CONCLUSIONS: An antibacterial peptide produced by Bacillus licheniformis was characterized, presenting a broad spectrum of activity against pathogenic and spoilage organisms . SIGNIFICANCE AND IMPACT OF THE STUDY: The identification of a substance active against important pathogens addresses an important aspect of food safety.

SADJ, 2004 Apr, 59(3), 119 - 22
Pulpitis induction in baboon primary teeth using carious dentine or Streptococcus mutans; Cleaton-Jones P et al.; The objective of this equivalency study was to see if a colony of Streptococcus mutans placed into cavities in primary molar teeth produced pulpitis similar to an established pulpitis induction method using carious dentine . In two juvenile baboons (Papio ursinus), occlusal cavities were cut in all 16 primary molar teeth, followed by making a small pulpal exposure after which the cavity was swabbed with 37 per cent phosphoric acid . In one half of the teeth, fresh soft human carious dentine was placed over the pulpal exposure; in the remaining teeth the exposure was covered with a colony of Streptococcus mutans in agar . All the cavities were restored with unlined light-cured composite resin . After 14 days specimens were harvested and examined under the light microscope with the examiner blind to the induction method . In both groups of teeth there was recognisable pulp, hyperaemia, micro-abscesses in the pulp and peri-apical abscesses . Reactions to soft caries were more severe than to Streptococcus mutans . The results show that Streptococcus mutans placed in a cavity with an exposure produces comparable pulpitis to fresh soft human carious dentine in the same type of cavity and that both methods produce pulpitis suitable for testing pulpotomy or pulpectomy treatments.

Acta Crystallogr D Biol Crystallogr, 2004 Jul, 60(Pt 7), 1341 - 5 Epub 2004 Jun 22.
Twinned or not twinned, that is the question: crystallization and preliminary crystallographic analysis of the 2F1(3)F1 module pair of human fibronectin; Rudino-Pinera E et al.; Human fibronectin (Fn) is a large multidomain protein found in the extracellular matrix and plasma . It is involved in many cellular processes, including cell adhesion and migration during embryogenesis and wound healing . The ability to bind Fn is a characteristic that has been demonstrated for a number of pathogens . For Staphylococcus aureus and Streptococcus pyogenes in particular, Fn-binding bacterial proteins (FnBPs) have been shown to mediate not only bacterial adhesion to host cells but also the uptake of bacteria by the cells . FnBPs interact with the amino-terminal region of Fn, where five type I ((1-5)F1) Fn modules are located . Although the structures of two F1 module pairs have been determined by NMR, no X-ray structures have been reported . To explore the conformational interactions between modules and the binding properties of FnBPs, the (2)F1(3)F1 module pair was crystallized using the vapour-diffusion method at 298 K . 12 X-ray diffraction data sets have been collected: six on an in-house rotating anode (three native, one Pt derivative and two peptide-bound) and six at synchrotron-radiation sources (two native and four derivative) . Following analysis of these data, some of which have very high multiplicity (up to 50), probable space-group assignments were made (P42(1)2, P4(1)2(1)2 or P4(3)2(1)2) and the possibly twinned nature of the crystals was investigated using six different tests . The results presented here suggest that the crystals are not twinned.

Acta Crystallogr D Biol Crystallogr, 2004 Jul, 60(Pt 7), 1266 - 71 Epub 2004 Jun 22.
Structure of the putative DNA-binding protein SP_1288 from Streptococcus pyogenes; Oganesyan V et al.; The crystal structure of the putative DNA-binding protein SP_1288 (gi/15675166, also listed as gi/28895954) from Streptococcus pyogenes has been determined by X-ray crystallography to a resolution of 2.3 A using anomalous diffraction data at the Se peak wavelength . SP_1288 belongs to a family of proteins whose cellular function is associated with the signal recognition particle; no structural information has been available until now about the members of the family . Crystallographic analysis revealed that the overall fold of SP_1288 consists exclusively of alpha-helices and that 75% of the structure has good similarity to domain 4 of the sigma subunit of RNA polymerase . This suggests its possible involvement in the biochemical function of transcription initiation, which includes interaction with DNA.

J Am Soc Nephrol, 2004 Jul, 15(7), 1785 - 93
Nephritis-associated plasmin receptor and acute poststreptococcal glomerulonephritis: characterization of the antigen and associated immune response; Yoshizawa N et al.; The role of nephritis-associated antigen as a virulence factor for acute poststreptococcal glomerulonephritis (APSGN) remains to be fully clarified . Nephritis-associated plasmin receptor (NAPlr) was previously isolated from group A streptococcus (GAS) and shown to bind plasmin(ogen) . The nucleotide sequence of the naplr gene from GAS isolates obtained from patients with APSGN was determined . The sequence of the putative open reading frame (1011 bp) showed 99.8% identity among isolated strains . Homology screen revealed an exact match with streptococcal glyceraldehyde-3-phosphate dehydrogenase (GAPDH) . NAPlr exhibited GAPDH activity in zymography, and it activated the complement pathway in vitro . In APSGN kidney biopsy specimens, NAPlr was observed mainly in the early stage of the disease (1 to 14 d after onset) but was not colocalized with either C3 or IgG as assessed by double immunofluorescence staining . Sera of patients with APSGN, patients with GAS infection without renal involvement, nonrenal pediatric patients, and healthy adults as controls were assayed for anti-NAPlr antibody titers . Anti-NAPlr antibodies were present most frequently in APSGN sera, and antibody titers were also significantly higher than in patients with GAS infection alone or in other control patients . Moreover, antibody titers remained elevated during the entire 10-yr follow-up period.

Infect Immun, 2004 Jul, 72(7), 4314 - 7
Identification of a point mutation resulting in loss of cell wall anchoring activity of SrtA of Streptococcus mutans NG5; Lee SF et al.; Streptococcus mutans NG5 failed to anchor antigen P1 to the cell surface, and such a failure could be attributed to a defective SrtA, which was made defective by a point mutation within the srtA gene . Without a functional SrtA, S . mutans NG5 was not able to perform a number of cell surface-related activities, including saliva-mediated adherence and aggregation.

Infect Immun, 2004 Jul, 72(7), 4309 - 13
Immunization with native or recombinant Streptococcus pneumoniae neuraminidase affords protection in the chinchilla otitis media model; Long JP et al.; Streptococcus pneumoniae neuraminidase has been implicated as a virulence factor in the pathogenesis of pneumococcal otitis media . In this study, native neuraminidase was partially purified from cultures of S . pneumoniae by serial chromatography with DEAE-Sepharose and Sephacryl S-200 . Recombinant neuraminidase, a 3,038-bp fragment of the neuraminidase A (nanA) gene, was cloned into the pET-28b vector and then expressed at high levels in Escherichia coli . Chinchillas were immunized subcutaneously with either the gel-purified native or recombinant neuraminidase, and all responded with elevated titers of antineuraminidase antibody in serum . Immunization with neuraminidase resulted in a significant reduction in nasopharyngeal colonization as well as in the incidence of otitis media with effusion . These data demonstrate for the first time that neuraminidase affords protection against S . pneumoniae nasopharyngeal colonization and experimental otitis media.

Infect Immun, 2004 Jul, 72(7), 4302 - 8
Function of the fibronectin-binding serum opacity factor of Streptococcus pyogenes in adherence to epithelial cells; Oehmcke S et al.; The serum opacity factor (SOF) of Streptococcus pyogenes is a serotyping tool and pathogenesis factor . Using SOF-coated latex beads in cell adherence assays and antiserum directed against SOF in S . pyogenes-HEp-2 cell adherence inhibition experiments, we demonstrate SOF involvement in the fibronectin-mediated adherence of S . pyogenes to epithelial cells . SOF exclusively targets the 30-kDa N-terminal region of fibronectin . The interaction revealed association and dissociation constants 1 order of magnitude lower than those of other S . pyogenes fibronectin-binding proteins.

Infect Immun, 2004 Jul, 72(7), 4290 - 2
Multiserotype protection of mice against pneumococcal colonization of the nasopharynx and middle ear by killed nonencapsulated cells given intranasally with a nontoxic adjuvant; Malley R et al.; Intranasal challenge of C57BL/6 mice with Streptococcus pneumoniae serotypes 6B, 14, and 23F produced colonization of the middle ear and NP . Intranasal vaccination with ethanol-killed nonencapsulated cells with adjuvant protected both sites . Of four nontoxic adjuvants tested, the cholera toxin B subunit was most effective and least nonspecifically protective.

Infect Immun, 2004 Jul, 72(7), 3981 - 6
Characterization of a complement-binding protein, DRS, from strains of Streptococcus pyogenes containing the emm12 and emm55 genes; Binks M et al.; An extracellular protein of Streptococcus pyogenes, streptococcal inhibitor of complement (SIC), and its variant, called DRS (distantly related to SIC), are expressed by some S . pyogenes strains . SIC from type 1 (M1) isolates of S . pyogenes interferes with complement-mediated cell lysis, reportedly via its interaction with complement proteins . In this study we demonstrate that S . pyogenes strains carrying emm12 and emm55 (the genes for the M12 and M55 proteins, respectively) express and secrete DRS . This protein, like SIC, binds to the C6 and C7 complement proteins, and competition enzyme-linked immunosorbent assay experiments demonstrate that DRS competes with SIC for C6 and C7 binding . Similarly, SIC competes with DRS for binding to the complement proteins . Despite this, the recombinant DRS preparation showed no significant effect on complement function, as determined by lysis of sensitized sheep erythrocytes . Furthermore, the presence of DRS is not inhibitory to SIC activity.

Infect Immun, 2004 Jul, 72(7), 3968 - 73
A two-component covRS regulatory system regulates expression of fructosyltransferase and a novel extracellular carbohydrate in Streptococcus mutans; Lee SF et al.; The expression of fructosyltransferase (FTF), the enzyme that synthesizes fructan from sucrose, is regulated in the cariogenic bacterium Streptococcus mutans . However, the exact mechanism of FTF regulation is unknown . In this study, the role of a two-component regulatory system (covRS) in FTF expression was investigated . A CovR-defective mutant of S . mutans NG8 was constructed by homologous recombination . By use of immunoblotting, the mutant was shown to overexpress FTF in the absence of sucrose, while the wild type and a covRS-complemented mutant showed sucrose-inducible FTF expression . Reverse transcription-PCR showed that the ftf transcript levels were increased in the covR mutant, suggesting regulation at the transcriptional level . The covR mutant was also found to overproduce extracellular carbohydrate, and this phenotype was reversed by covRS complementation . Paper chromatographic studies and chemical tests showed that the extracellular carbohydrate contained glucose and glucuronic acid but not fructose . These results suggest that the extracellular carbohydrate was not fructan . The production of a glucose- and glucuronic acid-containing extracellular carbohydrate has not been reported for S . mutans and may be considered novel . In conclusion, the results indicate that the expression of FTF and a glucose- and glucuronic acid-containing carbohydrate was negatively regulated by the covRS two-component regulatory system in S . mutans.

Infect Immun, 2004 Jul, 72(7), 3902 - 6
The Ami-AliA/AliB permease of Streptococcus pneumoniae is involved in nasopharyngeal colonization but not in invasive disease; Kerr AR et al.; The Ami-AliA/AliB oligopeptide permease is an ATP-binding cassette transporter which is found in Streptococcus pneumoniae and which is involved in nutrient uptake . We investigated the role of the three paralogous oligopeptide-binding lipoproteins AmiA, AliA, and AliB by using murine models of pneumococcal colonization and invasive disease . A series of mutants lacking aliA, aliB, and amiA either alone or in combination as double or triple mutations were used . Inoculation of the nasopharynx with a mixture of the obl (oligopeptide-binding lipoprotein-negative) triple-mutant and wild-type (D39) bacteria resulted in significantly smaller numbers of obl bacteria colonizing the nasopharynx . The use of a mixture of individual mutants and wild-type pneumococci revealed that AmiA, AliA, and AliB were all required for successful colonization of the nasopharynx . The obl mutant was more attenuated than the aliB mutant but not the aliA or amiA mutant . Therefore, there is some redundancy in the Ami-AliA/AliB complex in terms of nasopharyngeal colonization, with AliA and AmiA being able to compensate for the removal of AliB . Animals with invasive disease caused by these mutants had survival times, bacterial loads, and inflammatory cytokine production levels similar to those of animals infected with wild-type pneumococci . Our results show that although the Ami-AliA/AliB complex is not required for virulence during pneumococcal pneumonia, it does play a role in colonization of the nasopharynx.

Infect Immun, 2004 Jul, 72(7), 3876 - 82
Functional analysis of the Streptococcus gordonii DL1 sialic acid-binding adhesin and its essential role in bacterial binding to platelets; Takahashi Y et al.; Bacterial recognition of host sialic acid-containing receptors plays an important role in microbial colonization of the human oral cavity . The sialic acid-binding adhesin of Streptococcus gordonii DL1 was previously associated with the hsa gene encoding a 203-kDa protein . The predicted protein sequence consists of an N-terminal nonrepetitive region (NR1), including a signal sequence, a relatively short serine-rich region (SR1), a second nonrepetitive region (NR2), a long serine-rich region (SR2) containing 113 dodecapeptide repeats, and a C-terminal cell wall anchoring domain . In the present study, the contributions of SR1, NR2, and SR2 to Hsa-mediated adhesion were assessed by genetic complementation . Adhesion of an hsa chromosomal deletion mutant to sialic acid-containing receptors was restored by plasmids containing hsa constructs encoding Hsa that lacked either the N- or C-terminal portion of SR2 . In contrast, hsa constructs that lacked the coding sequences for SR1, NR2, or the entire SR2 region failed to restore adhesion . Surface expression of recombinant Hsa was not affected by removal of SR1, NR2, or a portion of SR2 but was greatly reduced by complete removal of SR2 . Wheat germ agglutinin, a probe for Hsa-specific glycosylation, reacted with recombinant Hsa lacking SR1, NR2, or SR2 but not with recombinant Hsa lacking both SR1 and SR2 . Significantly, the aggregation of human platelets by S . gordonii DL1, an interaction implicated in the pathogenesis of infective endocarditis, required the expression of hsa . Moreover, neuraminidase treatment of the platelets eliminated this interaction, further supporting the hypothesis that Hsa plays an essential role in the bacterium-platelet interaction.

Tex Heart Inst J, 2004, 31(2), 165 - 7
Endovascular treatment of intracerebral mycotic aneurysm before surgical treatment of infective endocarditis; Erdogan HB et al.; Mycotic aneurysms are rarely seen in patients who have infective endocarditis, and the management of these patients remains controversial . We present the case of a patient who had infective endocarditis complicated by a mycotic aneurysm of the left middle cerebral artery . There was substantial mitral regurgitation, and Streptococcus viridans was isolated from the blood samples . Dysarthria appeared during the 4th week of the antibiotic therapy, but resolved completely 8 hours after onset . The left middle cerebral artery was embolized with platinum detachable coils . On the 7th day after the radiologic intervention, the native mitral valve was replaced with a 33-mm St . Jude Medical bi-leaflet mechanical mitral prosthesis . Most mycotic aneurysms show notable regression of symptoms with effective antibiotic treatment, and a very few may diminish in size . However it is impossible to predict the response of these aneurysms to therapy . To prevent the perioperative rupture of mycotic aneurysms and intracranial hemorrhage, priority should be given to endovascular interventions to treat cerebrovascular aneurysms in patients such as ours.

Kansenshogaku Zasshi, 2004 May, 78(5), 446 - 50
{A case of invasive group A Streptococcus infection which was successfully treated with linezolid}; Nakamura S et al.; A 53-year-old male was admitted to our hospital with a high fever and pain in the right arm . He was diagnosed as toxic shock-like syndrome by Streptococcus pyogenes . His arm was amputated because of necrotizing myositis and his renal damage was severe, he was treated in the intensive care unit with continuous hemodiafiltration . Bacteria were isolated from blood, ascites, pleural effusion, skin, and muscle . He was treated with a large amount of ampicillin, clindamycin, and gammaglobulin . However, his general status became worse . His illness improved after linezolid was administered . The reason for the success in treatment with linezolid, which was the inhibitory effect on bacterial toxin and its excellent penetration into the tissue.

Kansenshogaku Zasshi, 2004 May, 78(5), 428 - 34
{Molecular epidemiology of fluoroquinolone-resistant Streptococcus pneumoniae in Japan}; Yokota S et al.; We identified fluoroquinolone-resistant Streptococcus pneumoniae strains among 670 clinical isolates isolated from 1999 to 2003 in Hokkaido prefecture, Japan . All eleven stains were resistant to ciprofloxacin and levofloxacin . Furthermore, ten strains were also resistant to fluoroquinolones that are more effective with gram-positive bacteria, namely tosufloxacin, sparfloxacin, and gatifloxacin . Nucleotide sequence analysis of the quinolone-resistance determining region (QRDR) of the quinolone target genes coding for topoisomerase i.v . subunits (parC and parE) and DNA gyrase subunits (gyrA and gyrB) . Eight stains, which showed higher resistance, had resistance mutations in two genes (gyrA and parC, or gyrA and parE), and other three strains had one resistance mutation in parC . The mutation patterns were varied between the strains . Data from random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) indicated that eleven strains were identified as ten independent clones . Lines of evidence indicated that genetic mutations leading to fluoroquinolone resistance occur sporadically rather through the spreading of a particular resistant strain . Notably, the fluoroquinolone-resistant strains were only isolated from adults, particularly from patients more than 60 years of age (9/60 strains; 15.0%) . Resistant strains were not found in 574 strains isolates from patients under 20 years of age . This may be due to the fact that fluoroquionolones other than norfloxacin are not applicable to children in Japan.

Chemotherapy, 2004 Jun, 50(2), 98 - 100
Comparative activities of beta-lactam antibiotics and quinolones for invasive Streptococcus pneumoniae isolates; Oncu S et al.; Background: Streptococcus pneumoniae is a leading pathogen causing pneumonia, meningitis, otitis media, bacteremia and sinusitis resulting in significant morbidity and mortality . We examined in vitro activities of five quinolones in comparison with other antibiotics against 85 invasive pneumococcal isolates . Methods: Minimal inhibitory concentrations (MICs) of penicillin G, cefuroxime, azithromycin, clarithromycin, trimethoprim-sulfamethoxazole (SXT), ciprofloxacin, ofloxacin, levofloxacin, trovafloxacin and gemifloxacin were determined using a broth microdilution method . Results: The overall rates of resistance to penicillin (46%), cefuroxime (20%), azithromycin (20%), clarithromycin (18%) and SXT (46%) were considerable . Among all of the isolates, 9 isolates (11%) were highly resistant (MIC >/=2 mg/l) and 30 isolates (35%) had intermediate resistance (MIC 0.12- 1.0 mg/l) . Of the quinolones gemifloxacin and trovafloxacin had the highest activity . The penicillin resistance status of the isolates did not have any effect on the resistance pattern of new quinolones . Conclusion: The new quinolones show great potential in the treatment of invasive infections caused by both penicillin-susceptible and penicillin-resistant pneumococci .

Chemotherapy, 2004 Jun, 50(2), 63 - 6
Differential efficacy of clarithromycin in lung versus thigh infection models; Maglio D et al.; BACKGROUND: Differences in clarithromycin disposition and the resulting changes in bacterial density were studied using mouse lung and thigh infection models . METHODS: Clarithromycin activity was evaluated against seven Streptococcus pneumoniae isolates with efflux-mediated resistance in both murine lung and thigh infection models . Intrapulmonary disposition of clarithromycin was also studied . RESULTS: Consistent bacterial kill was observed in the lung model, whereas no drug effect was observed in the thigh model . CONCLUSION: These differences in bacterial density were supported by high concentrations observed in epithelial lining fluid as compared to serum .

J Immunol, 2004 Jul 1, 173(1), 471 - 7
Dual roles of PspC, a surface protein of Streptococcus pneumoniae, in binding human secretory IgA and factor H; Dave S et al.; Streptococcus pneumoniae, also known as the pneumococcus, contains several surface proteins that along with the polysaccharide capsule function in antiphagocytic activities and evasion of the host immune system . These pneumococcal proteins interact with the host immune system in various ways and possess a wide range of biological activities that suggests that they may be involved at different stages of pneumococcal infection . PspC, also known as CbpA and SpsA, is one of several pneumococcal surface proteins that binds host proteins, including factor H (FH) and secretory IgA (sIgA) via the secretory component . Previous work by our laboratory has demonstrated that PspC on the surface of live pneumococcal cells binds FH . This paper provides evidence that FH activity is maintained in the presence of PspC and that the PspC binding site is located in the short consensus repeat 6-10 region of FH . We also report for the first time that although both FH and sIgA binding has been localized to the alpha-helical domain of PspC, the binding of FH to PspC is not inhibited by sIgA . ELISA, surface plasmon resonance, and flow cytometry indicate that the two host proteins do not compete for binding with PspC and likely do not share the same binding sites . We confirmed by Western analysis that the binding sites are separate using recombinant PspC proteins . These PspC variants bind FH yet fail to bind sIgA . Thus, we conclude that FH and sIgA can bind concurrently to the alpha-helical region of PspC.

Arch Dis Child, 2004 Jul, 89(7), 604 - 10
Dyskinesias and associated psychiatric disorders following streptococcal infections; Dale RC et al.; BACKGROUND: The classical extrapyramidal movement disorder following beta haemolytic streptococcus (BHS) infection is Sydenham's chorea (SC) . Recently, other post-streptococcal movement disorders have been described, including motor tics and dystonia . Associated emotional and behavioural alteration is characteristic . AIMS: To describe experience of post-streptococcal dyskinesias and associated co-morbid psychiatric features presenting to a tertiary referral centre 1999-2002 . METHODS: In all patients, dyskinetic movement disorders followed BHS pharyngeal infection . BHS infection was defined by pharyngeal culture of the organism, or paired streptococcal serology . Movement disorders were classified according to international criteria, and validated by experienced child neurologists . Psychiatric complications were defined using ICD-10 criteria using a validated psychiatric interview . RESULTS: In the 40 patients, the following dyskinetic movement disorders were present: chorea (n = 20), motor tics (n = 16), dystonia (n = 5), tremor (n = 3), stereotypies (n = 2), opsoclonus (n = 2), and myoclonus (n = 1) . Sixty five per cent of the chorea patients were female, whereas 69% of the tic patients were male . ICD-10 psychiatric diagnoses were made in 62.5% . Using the same psychiatric instrument, only 8.9% of UK children would be expected to have an ICD-10 psychiatric diagnosis . Emotional disorders occurred in 47.5%, including obsessive-compulsive disorder (27.5%), generalised anxiety (25%), and depressive episode (17.5%) . Additional psychiatric morbidity included conduct disorders (27.5%) and hyperkinetic disorders (15%) . Psychiatric, movement, and post-streptococcal autoimmune disorders were commonly observed in family members . At a mean follow up of 2.7 years, 72.5% had continuing movement and psychiatric disorders . CONCLUSION: Post-streptococcal dyskinesias occur with significant and disabling psychiatric co-morbidity and are potential autoimmune models of common "idiopathic" movement and psychiatric disorders in children . Multiple factors may be involved in disease expression including genetic predisposition, developmental status, and the patient's sex.

Oral Microbiol Immunol, 2004 Aug, 19(4), 257 - 61
Plasminogen interaction and activation on Streptococcus mutans surface; Urdaneta L et al.; A number of pathogenic microorganisms have been previously shown to bind plasminogen . The subsequent activation of plasminogen into plasmin can contribute to their virulence . In this study, we have shown that Streptococcus mutans is able to bind both human plasminogen and plasmin . Binding of plasminogen to S . mutans was inhibited by L-lysine and epsilon-aminocaproic acid, indicating that binding is mediated via lysine-binding sites of plasminogen . S . mutans enhanced the activation of plasminogen by tissue plasminogen activator but not by urokinase . This enhancement turned out to be dependent on cell concentration . Zymogram analysis showed that the plasmin activity acquired after plasminogen binding and activation is the most important proteolytic activity in the strain tested . These results suggest a mechanism involving acquisition of a host protease that might contribute to the infective process of this microorganism.

Oral Microbiol Immunol, 2004 Aug, 19(4), 240 - 6
Efficacy of ozone on survival and permeability of oral microorganisms; Nagayoshi M et al.; In the present study, we examined the effect of ozonated water on oral microorganisms and dental plaque . Almost no microorganisms were detected after being treated with ozonated water (4 mg/l) for 10 s . To estimate the ozonated water-treated Streptococcus mutans, bacterial cells were stained with LIVE/DEAD BacLight Bacterial Viability Kit . Fluorescence microscopic analysis revealed that S . mutans cells were killed instantaneously in ozonated water . Some breakage of ozonated water-treated S . mutans was found by electron microscopy . When the experimental dental plaque was exposed to ozonated water, the number of viable S . mutans remarkably decreased . Ozonated water strongly inhibited the accumulation of experimental dental plaque in vitro . After the dental plaque samples from human subjects were exposed to ozonated water in vitro, almost no viable bacterial cells were detected . These results suggest that ozonated water should be useful in reducing the infections caused by oral microorganisms in dental plaque.

Mol Divers, 2004, 8(2), 121 - 6
The rational design of an anti-caries peptide against Streptococcus mutans; Younson J et al.; The cariogenic bacterium Streptococcus mutans attaches to tooth surfaces via a cell surface adhesin termed streptococcal antigen I/II (SA I/II) . Mapping studies identified an adhesion epitope within residues 1025-1044 . A synthetic peptide (p1025) spanning these residues inhibited adhesion of S . mutans in vitro and was tested in an in vivo human streptococcal adhesion model . Direct application of p1025 to the teeth prevented recolonisation of the oral cavity by S . mutans but not Actinomyces naeslundii . This review also describes various other adhesion-inhibiting peptides have been identified in vitro . We suggest that adhesion-blocking synthetic peptides may provide novel anti-infective agents . Topical application of such peptides at mucosal surfaces does not provide sustained selective pressure and in contrast to antibiotics, may not induce resistance.

Mol Divers, 2004, 8(2), 113 - 20
Peptide antagonists of superantigen toxins; Kaempfer R; Superantigens produced by Staphylococcus aureus and Streptococcus pyogenes are among the most lethal of toxins . Toxins in this large family trigger an excessive cellular immune response leading to toxic shock . Superantigens are secreted by the bacteria as diverse natural mixtures, a complexity that demands development of broad-spectrum countermeasures . We used a rational approach to design short peptides with homology to various domains in a typical superantigen (staphylococcal enterotoxin B) and screened each peptide for its ability to antagonize, in human peripheral blood mononuclear cells, superantigen-mediated induction of the genes encoding T helper 1 cytokines that mediate shock: interleukin-2, interferon-gamma and tumor necrosis factor . A dodecamer peptide proved a potent antagonist against widely different superantigens . This peptide protected mice from killing by superantigens and it was able to rescue mice undergoing toxic shock . The antagonist peptide shows homology to a beta-strand-hinge-alpha-helix domain that is structurally conserved among superantigens, yet currently of unknown function and remote from the binding sites for the known ligands essential for T cell activation, the major histocompatibility complex class II molecule and T cell receptor . The antagonist activity of this peptide thus identifies a novel domain in superantigens that is critical for their toxic action . The antagonist peptide provides a new tool for understanding the mechanism of excessive human immune response activation by superantigens that occurs during toxic shock and for identification of a novel target ligand that may interact with this superantigen domain.

Wei Sheng Yan Jiu, 2004 Mar, 33(2), 216 - 8
{Protection of tooth protecting spray against dental carries}; Li Y et al.; OBJECTIVE: To investigate the growth, adherence and soul production of streptococcus mutans incubated with olitin tooth protecting spray and the effect of specific sample against dental caries . METHODS: Streptococcus mutans were cultured with the sample of Olitin tooth protecting spray . After culturing, S . mutans were observed by Gram stain and by spectrophotometer, and the percentages of the adherent S . mutans were calculated . 40 Wistar rats were divided into 4 groups: group A was control group; group B,C,D was low, middle, high test group respectively . All test groups received samples . The sum of the caries scores was assessed by the Keyes' procedure . RESULTS: The sample did not inhibit the growth of S . mutans, but the long chain was occurred . The adherence of S . mutans cells on the glass was decrease, the pH value was increase to above 6.0 . There was a significant lower mean of caries score of test group than control group . CONCLUSION: The sample can prevent the development of dental caries in this experiment . The mechanism may relate with the decrease of adherence of S . mutans.

Vet Immunol Immunopathol, 2004 Aug, 100(3-4), 145 - 9
The exploitation of the genome in the search for determinants of virulence in Streptococcus uberis; Leigh JA et al.; Despite much success in the control of mastitis in dairy cattle, intramammary infection with Streptococcus uberis remains a threat to herd health . This organism is a frequent cause of mastitis worldwide . Recent advances in the ability to genetically manipulate this bacterium, coupled to the determination of a representative genome sequence have already enabled the investigation of certain aspects of disease pathogenesis . Further use of such technology coupled to reliable models of disease and post-genomic analysis will permit the elucidation of further interactions between pathogen and host . This additional information can be usefully targeted at identification of candidates for inclusion in effective vaccines . This communication reviews the current, reported progress using this technology for S . uberis.

Rev Esp Med Nucl, 2004 Jul-Aug, 23(4), 282 - 3
{Psoas abscess as cause of lumbar spine pain detected by scintigraphy with gallium in a patient with suspicion of spondylodiscitis}; Ortega M et al.; A 56 year old man with fever and lumbar pain who underwent an abdominal CT scan that showed lumbar arthrosic changes, although it was not possible to rule out infectious disease in L5/S1 . Bone scintigraphy was requested . It showed heterogeneous hyperuptake that did not make it possible to exclude a spondylodiscitis in this site . Scintigraphy with 67Ga-citrate excluded infectious diseases in the lumbar spine column . However, a pathological uptake was observed in the left iliac fossa suggestive of psoas abscess, which was confirmed by ultrasonography, isolating streptococcus viridans.

Adv Exp Med Biol, 2003, 527, 67 - 76
Regulation of IDO-mediated bacteriostasis in macrophages: role of antibiotics and anti-inflammatory agents; MacKenzie CR et al.; Induction of IDO is also under strict control by the immune system and we have previously shown that there are a number of cytokines involved in the down-regulation of IDO induction . In clinical practice anti-inflammatory substances and antibiotics are commonly used and may influence the outcome of bacterial infection . We analysed the IFNgamma-dependant IDO induction and bacteriostasis of Staphylococcus aureus and Group A Streptococcus (GAS) in monocyte-derived-macrophages (MDM) from cord blood and peripheral blood of healthy adult donors with attention to the effect of down-regulatory cytokines and of two commonly used anti-inflammatory agents, hydrocortisone and indomethacin, on both IDO activity and bacterial growth . In addition to this we were interested in the effect of sub-inhibitory concentrations of the antibiotic ampicillin on this IDO-mediated effect, the premise being that for a substantial period of antibiotic therapy the infection site is exposed to sub-inhibitory concentrations of antibiotic . We found that after stimulation with IFNgamma, MDM inhibited streptococcal growth . This was due to IFNgamma-induced IDO activity as demonstrated by reconstitution of growth by supplemental tryptophan . This IDO-mediated bacteriostasis was inhibited by the cytokines IL-10, IL-4 and TGFbeta . Furthermore, addition of indomethacin to IFNgamma stimulated MDM also resulted in the abrogation of the IDO-induced bacteriostasis, a result of the inhibition of IDO induction . Surprisingly, co-stimulation with hydrocortisone and IFNgamma apparently increased the IDO activity in cord blood MDM, but had no effect on the IDO-activity of adult peripheral blood MDM . Bacteriostasis in cord blood MDM, on the other hand, was not affected by co-stimulation with hydrocortisone . Ampicillin, in sub-inhibitory concentrations had no effect on the IDO activity itself but did have a synergistic effect on the IDO-induced bacteriostasis in MDM cultures . We conclude that therapy with indomethacin may increase the risk of clinically important bacterial infection due to the inhibition of the IDO-induced bacteriostasis . In addition sub-inhibitory concentrations of ampicillin may play a role in the area of infection where IFNgamma stimulated macrophages are to be found in abundance.

J Bacteriol, 2004 Jul, 186(13), 4395 - 8
Characterization of Tn916S, a Tn916-like element containing the tetracycline resistance determinant tet(S); Lancaster H et al.; We have characterized a transferable tetracycline resistance (Tcr) element from a Streptococcus intermedius isolate . The gene responsible for this resistance was identified by PCR and Southern hybridization as tet(S) . Furthermore, the genetic support for this determinant was shown to be a conjugative transposon closely related to Tn916 . This element has been designated Tn916S.

J Bacteriol, 2004 Jul, 186(13), 4285 - 94
Multilocus sequence typing of Streptococcus pyogenes representing most known emm types and distinctions among subpopulation genetic structures; McGregor KF et al.; A long-term goal is to characterize the full range of genetic diversity within Streptococcus pyogenes as it exists in the world today . Since the emm locus is subject to strong diversifying selection, emm type was used as a guide for identifying a genetically diverse set of strains . This report contains a description of multilocus sequence typing based on seven housekeeping loci for 495 isolates representing 158 emm types, yielding 238 unique combinations of sequence type and emm type . A genotypic marker for tissue site preference (emm pattern) revealed that only 17% of the emm types displayed the marker representing strong preference for infection at the throat and that 39% of emm types had the marker for skin tropism, whereas 41% of emm types harbored the marker for no obvious tissue site preference . As a group, the emm types bearing the emm pattern marker indicative of no obvious tissue site preference were far less likely to have two distinct emm types associated with the same sequence type than either of the two subpopulations having markers for strong tissue tropisms (P < 0.002) . In addition, all genetic diversification events clearly ascribed to a recombinational mechanism involved strains of only two of the emm pattern-defined subpopulations, those representing skin specialists and generalists . The findings suggest that the population genetic structure differs for the tissue-defined subpopulations of S . pyogenes . The observed differences may partly reflect differential host immune selection pressures.

J Bacteriol, 2004 Jul, 186(13), 4152 - 8
The fabM gene product of Streptococcus mutans is responsible for the synthesis of monounsaturated fatty acids and is necessary for survival at low pH; Fozo EM et al.; Previously, it has been demonstrated that the membrane fatty acid composition of Streptococcus mutans is affected by growth pH (E . M . Fozo and R . G . Quivey, Jr., Appl . Environ . Microbiol . 70:929-936, 2004; R . G . Quivey, Jr., R . Faustoferri, K . Monahan, and R . Marquis, FEMS Microbiol . Lett . 189:89-92, 2000) . Specifically, the proportion of monounsaturated fatty acids increases when the organism is grown in acidic environments; if the shift to increased monounsaturated fatty acids is blocked by the addition of a fatty acid biosynthesis inhibitor, the organism is rendered more acid sensitive (E . M . Fozo and R . G . Quivey, Jr., Appl . Environ . Microbiol . 70:929-936, 2004) . Recently, work with Streptococcus pneumoniae has identified a novel enzyme, FabM, responsible for the production of monounsaturated fatty acids (H . Marrakchi, K . H . Choi, and C . O . Rock, J . Biol . Chem . 277:44809-44816, 2002) . Using the published S . pneumoniae sequence, a putative FabM was identified in the S . mutans strain UA159 . We generated a fabM strain that does not produce unsaturated fatty acids as determined by gas chromatography of fatty acid methyl esters . The mutant strain was extremely sensitive to low pH in comparison to the wild type; however, the acid-sensitive phenotype was relieved by growth in the presence of long-chain monounsaturated fatty acids or through genetic complementation . The strain exhibited reduced glycolytic capability and altered glucose-PTS activity . In addition, the altered membrane composition was more impermeable to protons and did not maintain a normal DeltapH . The results suggest that altered membrane composition can significantly affect the acid survival capabilities, as well as several enzymatic activities, of S . mutans.

Zhonghua Yi Xue Za Zhi, 2004 May 2, 84(9), 754 - 9
{Immunization with targeted fusion anticaries DNA vaccine via intramuscular route:experiment with murine}; Yu F et al.; OBJECTIVES: To observe the expression of a targeted fusion anticaries DNA vaccine pGJA-P in muscular in vivo . To compare the levels of specific antibodies and anticaries efficacy generated by pGJA-P and fusion anticaries DNA vaccine pGLUA-P in gnotobiotic rats, and observe the kinetics of antibody responses in BALB/c mice . METHODS: (1) Twelve 28-day-old female Wistar rats were randomly divided into 2 groups of 6 rats to be injected with the plasmid pGJA-P containing the signal peptide and extracellular regions of human CTLA(4), hinge and Fc regions of human IgG, the glu sequence of gtfB gene and A-P fragment of pac gene of Streptococcus mutans or the eukaryotic expression plasmid pCI into the quadriceps muscle of thigh respectively . Three days after the rats were killed and specimens of quadriceps muscles of thigh were taken . Immunohistochemical SP staining was used to examine the in situ expression of pGJA-P . (2) Twenty-four 18-day-old female Wistar rats were randomly divided into 4 groups of 6 rats . The rats were fed with cariogenic food . During the age of 20 - 22 days cariogentic food containing broad-spectrum antibiotics was fed . Then aseptic cotton stick was used to swab the oral cavity and be smeared onto the solid medium so as to observe the growth of bacteria under anaerobic culture for 48 hours . During the age of 24 - 26 days, S . mutans Ingbritt cultured anaerobically was swab onto the surface of teeth of the rats twice with an interval of 30 minutes . After the inoculation aseptic cotton stick was used to wipe the oral cavity and be smeared onto the solid medium so as to observe the growth of bacteria under anaerobic culture for 48 hours . When the gnotobiotic rats were 28 days old they were injected with pGJA-P, pGLUA-P, fusion anticaries DNA vaccine against both PAc, cell surface protein antigen, and glucosyltransferase (GTF), pCI or normal saline into the quadriceps muscle of thigh respectively, 2 weeks later a booster shot was given . When the rats were 63 days old their saliva and blood samples were collected . The serum IgG and salivary IgA were assayed by using ELISA . The gnotobiotic rats were killed and their maxillary bone the mandibles were isolated . The anticaries effect was evaluated by Keyes caries scores . (3) Twenty-four 4-week-old BALB/c mice were randomly divided into 4 groups of 6 mice: to be injected with pGJA-P, pGLUA-P, pCI, or normal saline respectively into the quadriceps muscles of thigh, 2 weeks later a booster shot was given . Before the injection and every 2 weeks after the immunization specimens of saliva and blood were collected . The serum IgG and salivary IgA were assayed by using ELISA . RESULTS: (1) Recombinant protein could be detected in the quadriceps muscles of the rats immunized with pGJA-P, but not in the muscles of the rats immunized with pCI . (2) The levels of serum anti-PAc IgG (1:200 000) and anti-GTF IgG (1:58 000) of the rats immunized with pGJA-P were significantly higher than those of the rats immunized with pGLUA-P (1:23 000 and 1:11 000 respectively) (both P < 0.01) . The levels of salivary anti-PAc IgA (1:8) and anti-GTF IgA (1:6) of the rats immunized with pGJA-P were significantly higher than those of the rats immunized with pGLUA-P (1:2 and 1:2 respectively) (both P < 0.01) . The Keyes scores of the pGJA-P group were significantly lower than those of the pGLUA-P group and the control groups (all P < 0.01) . The effective serum IgG and salivary IgA in the pGJA-P group and effective serum IgG in the pGLUA-P group all persisted to the end of the experiment . (3) Two weeks after the initial immunization the serum anti-PAc IgG level of the mice immunized with pGJA-P increased remarkably, 4 times that of the mice immunized with pGLUA-P, and 33 times those of the mice injected with pCI or normal saline . Two weeks after the booster immunization, the serum anti-PAc IgG level of the mice immunized with pGJA-P was 14 times that of the mice immunized with pGLUA-P, and 117 times those of the mice injected with pCI or normal saline . The serum anti-PAc IgG immunized with pGJA-P reached its peak 10 weeks after the initial immunization, 4 times that of the mice immunized with pGLUA-P, and 160 times those of the mice injected with pCI or normal saline . The serum anti-PAc IgG of the mice immunized with pGLUA-P reached its peak at 16 weeks, however, significantly lower than the peak of the mice immunized with pGJA-P (P < 0.01) . The serum anti-Pac IgG levels of the mice injected with pCI or with normal saline were not significantly different (P > 0.05) . Since the second week after the initial immunization, significant difference in the serum anti-PAc IgG level could be seen between the mice immunized with pGJA-P or the mice immunized with pGLUA-P, and between the mice immunized with pGJA-P and the mice immunized with pGLUA-P and those injected with pCI or normal saline (all P < 0.01) . Six weeks after the initial immunization the salivary anti-PAc IgA level of the mice immunized with pGJA-P was 18 times those of the mice injected with pCI or with normal saline (both P < 0.01), 10 weeks after the salivary anti-PAc IgA level of the mice immunized with pGJA-P reached its peak, 24 times those of the mice immunized with pCI or normal saline without a significant difference between the latter 2 groups (P > 0.05) . No effective salivary IgA response was seen in the mice immunized with pGLUA-P . CONCLUSION: pGJA-P can be expressed in vivo . Immunization with pGJA-P intramuscularly induces effective mucosal and systematic humoral responses . It is an effective DNA vaccine against dental caries.

Vet Radiol Ultrasound, 2004 May-Jun, 45(3), 210 - 5
Magnetic resonance imaging of a brain abscess in a 10-month-old filly; Audigie F et al.; The purpose of this paper was to correlate the magnetic resonance imaging (MRI) characteristics of a mature brain abscess in a horse with histopathologic alterations of brain tissue . Eight months after the onset of clinical signs, MRI of the brain of a 10-month-old filly was performed . A large space-occupying lesion in the right cerebral hemisphere was identified . This space-occupying lesion was delineated by a thick and well-defined capsule that was isointense to brain parenchyma on the T1-weighted images and with a markedly hypointense on the T2-weighted images . The identification of such a capsule is highly diagnostic of a mature brain abscess . The lesion seen on MR images was confirmed at necropsy where a large abscess of the right hemisphere was observed . Streptococcus zooepidemicus and Pseudomonas aeruginosa were isolated from the abscess . Based on histopathologic examination, the signal characteristics of the capsule on T1-weighted and T2-weighted images were found to be due to the presence of numerous hemosiderin-laden macrophages . These results are in agreement with previous studies on human patients . This report confirms the value of MRI in the diagnosis of equine brain diseases.

Eur J Pediatr, 2004 Sep, 163(9), 509 - 16 Epub 2004 Jun 10.
What can children gain from pneumococcal conjugate vaccines?
Peltola H, Booy R, Schmitt HJ.
In excess of 1 million young children die every year as a consequence of disease caused by Streptococcus pneumoniae, the vast majority in developing countries . Although the first vaccine against the Pneumococcus was produced before the First World War, licensure of the first vaccine with documented efficacy against severe infections in infants and young children did not occur until February 2000 in the United States . This conjugate vaccine consists of purified polysaccharide, from each of seven pneumococcal serotypes, chemically linked to a carrier protein . A high degree of efficacy of the new vaccine against potentially life-threatening infections has been shown in both poor and affluent countries . The vaccine's potential to protect from acute otitis media, however, is very limited, although encouraging indirect effects, such as reduced antibiotic prescriptions, have been reported . An inherent problem with the new pneumococcal conjugate vaccines is that, while more than 20 pneumococcal serotypes may cause invasive disease, only a more limited number of polysaccharides, 11 or so, can in practice be conjugated to carrier protein as part of a single vaccine formulation . Because of variation in the ranking of serotypes most commonly responsible for pneumococcal disease, by region, age and disease manifestation, compromise was required in selecting serotype-specific saccharides for inclusion . CONCLUSION: Complex conjugate technology comes at a price, and the present costs keep most of the world's children far out of reach of an effective vaccine . However, the pneumococcal conjugate vaccine is a highly functional weapon against deadly pneumococcal infections, and strenuous efforts are needed to maximise its accessibility to children most at risk.

Environ Int, 2004 Sep, 30(7), 911 - 22
Bioremediation of coastal areas 5 years after the Nakhodka oil spill in the Sea of Japan: isolation and characterization of hydrocarbon-degrading bacteria; Chaerun SK et al.; Five years after the 1997 Nakhodka oil spill in the Sea of Japan, seven bacterial strains capable of utilizing the heavy oil spilled from the Nakhodka Russian oil tanker were isolated from three coastal areas (namely Katano Seashore of Fukui Prefecture, Osawa and Atake seashores of Ishikawa Prefecture) and the Nakhodka Russian oil tanker after a 5-year bioremediation process . All bacterial strains isolated could utilize long-chain-length alkanes efficiently, but not aromatic, and all of them were able to grow well on heavy oil . Using 16S rDNA sequencing, most of the strains were affiliated to Pseudomonas aeruginosa . Comparing between the year 1997 (at the beginning of bioremediation process) and the year 2001 (after 5 years of bioremediation), there was no significant change in morphology and size of hydrocarbon-degrading bacteria during the 5-year bioremediation . Scanning and transmission electron microscopic observations revealed that a large number of hydrocarbon-degrading bacteria still existed in the sites consisting of a variety of morphological forms of bacteria, such as coccus (Streptococcus and Staphylococcus) and bacillus (Streptobacillus) . On the application of bioremediation processes on the laboratory-scale, laboratory microcosm experiments (containing seawater, beach sand, and heavy oil) under aerobic condition by two different treatments (i.e., placed the inside building and the outside building) were established for bioremediation of heavy oil to investigate the significance of the role of hydrocarbon-degrading bacteria on them . There was no significant bacterial activity differentiation in the two treatments, and removal of heavy oil by hydrocarbon-degrading bacteria in the outside building was slightly greater than that in the inside building . The values of pH, Eh, EC, and dissolved oxygen (DO) in two treatments indicated that the bioremediation process took place under aerobic conditions (DO: 1-6 mg/l; Eh: 12-300 mV) and neutral-alkaline conditions (pH 6.4-8) with NaCl concentrations of 3-15% (ECs of 45-200 mS/cm).

Clin Exp Immunol, 2004 Jul, 137(1), 179 - 86
The impact of an early truncating founder ATM mutation on immunoglobulins, specific antibodies and lymphocyte populations in ataxia-telangiectasia patients and their parents; Stray-Pedersen A et al.; Eleven Norwegian patients (aged 2-33 years, seven males and four females) with Ataxia-telangiectasia (A-T) and their parents were investigated . Five of the patients were homozygous for the same ATM mutation, 3245delATCinsTGAT, a Norwegian founder mutation . They had the lowest IgG2 levels; mean (95% confidence interval) 0.23 (0.05-0.41) g/l versus 0.91 (0.58-1.26) g/l in the other patients (P = 0.002) . Among the 11 A-T patients, six had IgG2 deficiency, six had IgA deficiency (three in combination with IgG2 deficiency) and seven had low/undetectable IgE values . All patients had very low levels of antibodies to Streptococcus pneumoniae 0.9 (0.4-1.4) U/ml, while normal levels were found in their parents 11.1 (8.7-13.4) U/ml (P < 0.001) . A positive linear relationship between pneumococcal antibodies and IgG2 (r = 0.85, P = 0.001) was found in the patients . Six of 11 had diphtheria antibodies and 7 of 11 tetanus antibodies after childhood vaccinations, while 4 of 7 Hemophilus influenzae type b (Hib) vaccinated patients had protective antibodies . Ten patients had low B cell (CD19+) counts, while six had low T cell (CD3+) counts . Of the T cell subpopulations, 11 had low CD4+ cell counts, six had reduced CD8+ cell counts, and four had an increased portion of double negative (CD3+/CD4-/CD8-) gamma delta T cells . Of the 22 parents (aged 23-64 years) 12 were heterozygous for the ATM founder mutation . Abnormalities in immunoglobulin levels and/or lymphocyte subpopulations were also observed in these carriers, with no correlation to a special ATM genotype.

Clin Exp Immunol, 2004 Jul, 137(1), 35 - 40
Acute infection with influenza virus enhances susceptibility to fatal pneumonia following Streptococcus pneumoniae infection in mice with chronic pulmonary colonization with Pseudomonas aeruginosa; Seki M et al.; We established a mouse model in which fatal pneumonia was induced by pneumococcal superinfection following influenza virus infection in chronic Pseudomonas aeruginosa infected mice . In this mouse model, influenza virus infection caused a significant increase in inflammatory cells, cytokines and severe tissue damage in the lungs of these P . aeruginosa infected mice, before pneumococcal infection . Intrapulmonary virus titres were significantly increased in mice with chronic P . aeruginosa infection, compared with control mice . Neutrophil function analysis showed significant reduction of myeloperoxidase (MPO) activity and lysozyme secretion by influenza virus infection in these mice . Our results suggest that influenza virus infection may play an important role in inducing pneumococcal pneumonia in chronic P . aeruginosa infected mice . Our results suggested that our mouse model is useful for investigating the pathogenesis of influenza virus infection in patients with chronic lung infection.

Pediatr Infect Dis J, 2004 Jun, 23(6), 574 - 6
Aortitis in a child with Abiotrophia defectiva endocarditis; Raff GW et al.; Abiotrophia defectiva, one of several nutritionally variant Streptococcus species, is an uncommon but important cause of endocarditis in children . We describe an unusual case complicated by extensive aortitis with pits in the ascending aorta and the proximal aortic arch.

Vaccine, 2004 Jun 23, 22(19), 2362 - 7
Development of experimental carbohydrate-conjugate vaccines composed of Streptococcus pneumoniae capsular polysaccharides and the universal helper T-lymphocyte epitope (PADRE); Alexander J et al.; Experimental carbohydrate-conjugate vaccines composed of the 13 amino acid universal Pan HLA-DR Epitope (PADRE) and Streptococcus pneumoniae capsular polysaccharides from serotypes 14, 6B and 9V were produced . Simple carbodiimide-mediated condensation chemistry was used to conjugate the PADRE synthetic peptide to the three chemically different capsular polysaccharides in a 1:1 molar ratio . The immunogenicity of the PADRE peptide component of the conjugate vaccines was confirmed by the induction of PADRE-specific CD4+ helper T cell (HTL) responses following immunization of C57BL/6 mice . High titer antibody responses specific for polysaccharides of S . pneumoniae serotypes 14, 6B and 9V were induced using Complete Freund's Adjuvant (CFA) and alhydrogel Al(OH)3 formulations . The HTL, or carrier, effect of the PADRE synthetic peptide was only evident using the PADRE-polysaccharide conjugates; simple mixtures of the PADRE peptide and polysaccharides were essentially nonimmunogenic . The functional or potential protective value of the polysaccharide-specific antibodies was measured as a function of opsonophagocytic activity for the 6B serotype . High titers of opsonophagocytic activity were measured in sera from mice immunized with formulations containing both adjuvants . These data demonstrate that the PADRE synthetic peptide can induce the HTL responses needed to support the development of antibodies specific for bacterial carbohydrates used in conjugate vaccines.

Vaccine, 2004 Jun 30, 22(20), 2511 - 6
Mucosal immunization against dental caries with plasmid DNA encoding pac gene of Streptococcus mutans in rats; Jia R et al.; Salivary secretory immunoglobulin A (S-IgA) antibodies act as the first line of defense against dental caries by blocking of adherence of Streptococcus mutans to tooth surfaces . This study focused on finding proper mucosal immunization route and delivery system to induce higher level of specific anti-S . mutans saliva S-IgA and inhibit dental caries in animal model . By immunizing rats with an anti-caries DNA vaccine, pCIA-P, via different mucosal routes, we found that intranasal (i.n.) immunization with pCIA-P/bupivacaine DNA complexes elicited the highest specific anti-S . mutans saliva S-IgA mucosal antibody responses compared with naked DNA and other routes . Correspondingly, rats immunized with pCIA-P/bupivacaine DNA complex via i.n . displayed the least carious lesions . Our findings suggested that DNA vaccination via intranasal immunization with bupivacaine delivery system be a promising approach against dental caries.

Clin Microbiol Infect, 2004 Jun, 10(6), 587 - 9
CD4 T-lymphocyte subset counts in HIV-seropositive patients during the course of community-acquired pneumonia caused by Streptococcus pneumoniae; Schleicher GK et al.; Total lymphocyte counts, CD4 T-lymphocyte counts and CD4/CD8 ratios were measured in 30 anti-retroviral-naive HIV-seropositive patients upon hospital admission for acute community-acquired pneumonia (CAP) caused by Streptococcus pneumoniae, and again 1 month after resolution of infection . There was a significant depression of the total lymphocyte count (p < 0.005) and CD4 T-lymphocyte count (p < 0.001) in the acute stage of CAP caused by S . pneumoniae, with a subsequent increase in 90% (27/30) of cases after resolution of the infection . There was no significant difference in the CD4/CD8 T-lymphocyte ratio on admission compared with 1 month later (p 0.9).

Ann Allergy Asthma Immunol, 2004 May, 92(5), 558 - 64
Immunogenicity of 23-valent pneumococcal polysaccharide vaccine in children with human immunodeficiency virus undergoing highly active antiretroviral therapy; Tangsinmankong N et al.; BACKGROUND: The 23-valent pneumococcal polysaccharide vaccine (23PSV) has been recommended for children infected with human immunodeficiency virus (HIV); however, the efficacy of this vaccination in HIV-infected children undergoing highly active antiretroviral therapy (HAART) has not been studied . OBJECTIVE: To study the immunogenicity and immunologic protection of 23PSV in HIV-infected children after stable HAART . METHODS: Serotype-specific IgG antibodies to 12 pneumococcal capsular polysaccharides were analyzed before and after 23PSV vaccination in 41 HIV-infected children undergoing HAART and compared with 95 HIV-negative control children . Seropositivity, clinical protection, and additional clinical protection from 23PSV were calculated based on serotype specific IgG antibody levels and on the known incidence of these serotypes for causing invasive disease . RESULTS: Children with HIV infection undergoing HAART developed a significant increase in specific IgG levels to Streptococcus pneumoniae after 23PSV vaccination (0.95 vs 1.84 micro/gmL, P < .001) . The HIV-infected children with CD4+ cell counts of 25% or higher at the time of vaccination developed a higher additional clinical protection gain from 23PSV vaccination than did children with a lower percentage of CD4+ cells . CONCLUSIONS: HIV-infected children undergoing stable HAART develop a significant immunologic response to 23PSV, especially those with higher T-cell counts and lower viral loads at the time of vaccination.

J Dairy Res, 2004 May, 71(2), 175 - 81
Milk L-lactate concentration is increased during mastitis; Davis SR et al.; A study was undertaken in cattle to evaluate changes in milk L-lactate in relation to mastitis . A healthy, rear quarter of the udder of each of ten cows in mid-lactation was infused with 1000 colony-forming units (cfu) of Streptococcus uberis following an afternoon milking . Foremilk samples were taken at each milking from control and treated quarters and antibiotic treatment was applied following the onset of clinical mastitis or after 72 h . One cow did not become infected . Six quarters showed clinical symptoms of mastitis within 24-40 h and this was associated with a more than 30-fold increase in milk L-lactate (to 3.3 mM) and an increase in somatic cell count (SCC) from 4.5 x 10(3) to 1 x 10(7) cells/ml . Three cows were subclinical, with cell counts ranging from 1.5 x 10(6) to 1 x 10(7) cells/ml . In these animals, milk lactate ranged from 0.7 to 1.5 mM in the infected quarters up to 40 h post-infection, compared with less than 0.1 mM in control quarters . Milk was examined from 137 cows in mid-lactation which were known to have mastitis . Foremilk samples were taken aseptically from control and infected quarters of cows on commercial farms . Mean milk L-lactate concentrations and SCC were 0.14 +/- 0.02 mM and 1.85 +/- 0.3 x 10(5) cells/ml, respectively, in control (bacteriologically negative) samples . However, L-lactate concentrations exceeded 2.5 mM in the presence of some types of infection, the level of the lactate response being closely related to the impact of the infection on SCC . L-Lactate concentrations were relatively elevated in milk samples taken post partum, declining from 0.8 to 0.14 mM oyer the first few days of lactation . In conclusion, milk L-lactate has potential as an indicator of clinical and subclinical mastitis in dairy cows.

Hua Xi Kou Qiang Yi Xue Za Zhi, 2004 Apr, 22(2), 149 - 51
{Effect of para-aminobenzonic acid (PABA) on cell-surface hydrophobicity of Streptococcus mutans}; Li YH et al.; OBJECTIVE: To study the effect of Para-aminobenzonic acid on cell-surface hydrophobicity of Streptococcus mutans . METHODS: Microbial adhesion to hydrocarbons (MATH) was used to measure the cell-surface hydrophobicity of Streptococcus mutans which grew in modified Carlsson medium with different dilutions of PABA . RESULTS: The cell-surface hydrophobicity of Streptococcus mutans increased when Carlsson medium contained low dilution of PABA . But following the increase of PABA, the cell-surface hydrophobicity decreased . CONCLUSION: Para-aminobenzonic acid could inhibit the adherence of Streptococcus mutans through changing its cell-surface hydrophobicity.

Nucleic Acids Res, 2004 Jun 09, 32(10), 3136 - 47 Print 2004.
Recognition of DNA by omega protein from the broad-host range Streptococcus pyogenes plasmid pSM19035: analysis of binding to operator DNA with one to four heptad repeats; de la Hoz AB et al.; pSM19035-encoded omega protein forms a dimer (omega2) that binds to a set of 7-bp repeats with sequence 5'-NATCACN-3' . Upon binding to its cognate sites, omega2 regulates transcription of genes required for copy number control and stable inheritance of plasmids, and promotes accurate plasmid segregation . Protein omega2 binds poorly to one heptad but the affinity to DNA increases with two and more unspaced heptads in direct or inverted orientation . DNA titration of increasing numbers of heptads with omega2, monitored by circular dichroism measurements, indicates the binding of one omega2 to one heptad (omega2:heptad stoichiometry of 1:1) . Spacing of two directly or inversely oriented heptads by 1 to 7 bp reduces the affinity of the protein for its cognate target site . The binding affinity of omega2 for two directly repeated heptads was severely reduced if one of the base pairs of the core 5'-ATCAC-3' sequence of one of the heptads was individually substituted by any other base pair . Hydroxyl radical footprinting shows a protection pattern at the 5'-ATCAC-3' core . These data suggest that each heptad defines an operator half-site and that tight binding of the symmetric omega2 to the central 5'-TCA-3' core of symmetric or asymmetric targets (differently oriented heptads) is probably achieved by structural changes of DNA and/or protein or both.

J Biol Chem, 2004 Aug 6, 279(32), 33281 - 9 Epub 2004 Jun 09.
A novel mechanism for the inhibition of hyaluronan biosynthesis by 4-methylumbelliferone; Kakizaki I et al.; Specific inhibitors of hyaluronan (HA) biosynthesis can be valuable therapeutic agents to prevent cancer invasion and metastasis . We have found previously that 4-methylumbelliferone (MU) inhibits HA synthesis in human skin fibroblasts and in group C Streptococcus . In this paper, the inhibition mechanism in mammalian cells was investigated using rat 3Y1 fibroblasts stably expressing HA synthase (HAS) 2 . Exposure of the transfectants to the inhibitor resulted in significant reduction of HA biosynthesis and matrix formation . The evaluation of HAS transcripts and analysis of cell-free HA synthesis demonstrated the post-transcriptional suppression of HAS activity by MU . Most interesting, the post-transcriptional suppression of HAS activity was also observed using p-nitrophenol, a well known substrate for UDP-glucuronyltransferases (UGT) . We investigated whether the inhibition was exerted by the glucuronidation of MU using both high pressure liquid chromatography and TLC analyses . The production of MU-glucuronic acid (GlcUA) was consistent with the inhibition of HA synthesis in HAS transfectants . MU-GlcUA was also detected at a similar level in control cells, suggesting that the glucuronidation was mediated by an endogenous UGT . Elevated levels of UGT significantly enhanced the inhibitory effects of MU . In contrast, the inhibition by MU was diminished to the control level when an excess of UDP-GlcUA was added to the cell-free HA synthesis system . We propose a novel mechanism for the MU-mediated inhibition of HA synthesis involving the glucuronidation of MU by endogenous UGT resulting in a depletion of UDP-GlcUA.

J Antimicrob Chemother, 2004 Jul, 54(1), 76 - 8 Epub 2004 Jun 09.
The glycopeptide vancomycin does not enhance toll-like receptor 2 (TLR2) activation by Streptococcus pneumoniae; Moore LJ et al.; OBJECTIVES: The exposure of Streptococcus pneumoniae to cell-wall-active antibiotics in vivo and in vitro results in the release of bacterial components that can induce proinflammatory activation of human cells via toll-like receptor 2 (TLR2) . The aim of this study was to compare the activation of human TLR2 pathways after exposure of S . pneumoniae to faropenem, cefotaxime and vancomycin . MATERIALS AND METHODS: Streptococcus pneumoniae D39 was exposed to cefotaxime, faropenem or vancomycin for 6 h during lag or early log phase growth . IL-8 promoter activity of HeLa cells was measured using a dual luciferase reporter plasmid system . HeLa cells were transfected with an expression vector containing TLR2/CD14, or empty vector/CD14 and IL-8 promoter activity was measured using luminescence . Cells were stimulated with antibiotic-treated bacteria, untreated bacteria or medium-only controls . RESULTS: Lag phase S . pneumoniae treated at sub-MIC (1/8 MIC) cefotaxime or faropenem induced 11-fold and 8-fold increases, respectively, in TLR2-mediated IL-8 promoter activity when compared with untreated bacteria . Early log MIC cefotaxime or faropenem-treated bacteria also enhanced TLR2 activation by 3-fold and 4-fold, respectively, when compared with untreated bacteria . Vancomycin treatment had no effect on TLR2 induction at any growth stage or MIC ratio tested . CONCLUSIONS: beta-Lactam antibiotics induce surface changes and release of cell wall structures from bacteria that are proinflammatory via TLR2, but the glycopeptide vancomycin does not.

J Antimicrob Chemother, 2004 Jul, 54(1), 117 - 21 Epub 2004 Jun 09.
Phenotypic and molecular characterization of macrolide and streptogramin resistance in Streptococcus mitis from neutropenic patients; Achour W et al.; OBJECTIVES: To determine the prevalence of macrolide and streptogramin resistance in Streptococcus mitis isolates from neutropenic patients and to identify mechanisms of macrolide and streptogramin resistance in resistant isolates . METHODS: MICs of erythromycin, spiramycin, lincomycin and pristinamycin were determined for S . mitis isolates . Macrolide-resistance genes were characterized by PCR and ribosomal mutations by sequencing . RESULTS: A total of 169 S . mitis isolates were recovered from 66 patients at the Tunisian Bone Marrow Transplant Centre . Of these, 120 (70%) were non-susceptible to erythromycin and one was resistant to pristinamycin; 48.5% of isolates had an MLSB phenotype with cross-resistance between erythromycin, spiramycin and lincomycin, 4% had a dissociated MLSB phenotype with resistance to erythromycin and spiramycin but apparent susceptibility to lincomycin and 47.5% displayed the M phenotype . Resistance determinants were characterized in 33 isolates . Ten of 14 isolates with the cross MLSB resistance contained an ermB-like gene and four a combination of ermB- and mefA-like genes . Four of the five isolates with a dissociated MLSB phenotype contained ermB-like and one a combination of ermB- and mefA-like genes . All the 14 isolates with an M phenotype contained mefA-like genes . The pristinamycin-resistant strain had G105 and A108 substitutions in the conserved C terminus of the L22 ribosomal protein . CONCLUSIONS: The prevalence of macrolide resistance is high in S . mitis from neutropenic patients and is due to the spread of ermB- or mefA-like genes alone or combined . Resistance to streptogramins is rare and in this case associated with ribosomal mutation.

J Oral Rehabil, 2004 Jun, 31(6), 568 - 73
Effect of saliva on an antimicrobial tissue conditioner containing silver-zeolite; Abe Y et al.; The purpose of this study was to evaluate the influence of human saliva on the antimicrobial effect of a tissue conditioner containing an antibiotic agent, silver-zeolite . Samples of each tissue conditioner with or without silver-zeolite were prepared and a plastic disk was used as a control . Candida albicans and nosocomial respiratory infection-causing bacteria, Staphylococcus aureus, methicillin-resistant S . aureus (MRSA), Pseudomonas aeruginosa and the Streptococcus milleri group (S . constellatus and S . intermedius), were selected as test microorganisms . Antimicrobial effects of samples after water or saliva immersion for 28 days were evaluated by counting the number of viable cells {colony forming unit (CFU)} in each microbial suspension (100 microL) . All data were statistically analysed by one-way anova and Bonferroni's test (P < 0.05) . The antimicrobial effects of samples with silver-zeolite immersed in saliva against C . albicans, S . aureus and MRSA were observed while CFU of P . aeruginosa indicated no significant difference from that of the control . As for the S . milleri group, its CFU after saliva immersion showed the significantly smaller value than that of the control . It is concluded that the antimicrobial effects of samples containing silver-zeolite against all tested microbes except for P . aeruginosa and the S . milleri group are not influenced by saliva immersion for 28 days.

Lett Appl Microbiol, 2004, 39(1), 84 - 8
Molecular heterogeneity among north Indian isolates of Group A Streptococcus; Sagar V et al.; AIM: To monitor molecular heterogeneity among the clinical isolates of group A Streptococcus (GAS) from north India by Vir and emm typing . METHODS AND RESULTS: GAS isolates, 31 from pharyngitis and nine from rheumatic fever (RF)/rheumatic heart disease (RHD) patients were differentiated into 16 Vir types (VT) . These isolates were further discriminated into 23 emm types . Most of emm types were Vir type specific, except few (7.5%), which revealed different Vir types within same emm type . The most prevalent emm type found was emm 49 (15%) followed by 7.5% of emm 69, emm 71 and emm 75 which were different from emm type distribution reported from south India . CONCLUSIONS: Analysis of data revealed 40% heterogeneity by Vir typing and 57.5% by emm typing among GAS isolates which is significant in view of small number of isolates studied . SIGNIFICANCE OF IMPACT OF THE STUDY: The molecular study for the first time demonstrates different emm types prevalent and circulating in northern region of India and such data may help in selection of types for vaccine development.

Epidemiol Infect, 2004 Jun, 132(3), 433 - 41
Estimating the transmission parameters of pneumococcal carriage in households; Melegaro A et al.; This paper analyses Streptococcus pneumoniae transmission dynamics in households using longitudinal data on pneumococcal (Pnc) carriage in the United Kingdom . Ten consecutive swabs were taken at 4-week intervals from all members of 121 households . The family status is derived from the observed Pnc carriage status of each family member . Transition matrices are built for each family size and composition containing the observed frequency of transitions between family statuses over a 28-day interval . A density-dependent transmission model is fitted to derive maximum-likelihood estimates of the duration of carriage and acquisition rates from the community and from infected individuals within the household . Parameter values are estimated for children (< 5 years) and adults (5+ years) . The duration of carriage is longer in children < 5 years of age than in older family members (51 vs . 19 days) . Children are 3-4 times more likely than adults to acquire Pnc infection from the community . Transmission rates within the household suggest that adults are more infectious but less susceptible than children . Transmission within the household is most important in large families . The proportion of household-acquired infection ranges from 29 to 46% in households of three persons to 38-50% in larger households . Evidence of density-dependent within-household transmission is found, although the strength of this relationship is not clear from the model estimates.

Zh Mikrobiol Epidemiol Immunobiol, 2004 Mar-Apr, (2), 81 - 3
{Antibiotic resistance of Streptococcus pneumoniae isolated from patient with home care pneumonia}; Martynova AV et al.; The monitoring of the resistance of Streptococcus pneumoniae, one of the most important causative agents of extrahospital pneumonia, has been carried out . The data on its sensitivity to antibacterial preparations, widely used in clinical practice, have been summarized.

Infection, 2004 Jun, 32(3), 179 - 81
Legionella in two splenectomized patients . Coincidence or causal relationship?
Gorelik O, Lazarovich Z, Boldur I, Almoznino-Sarafian D, Alon I, Modai D, Cohen N.
We describe two splenectomized patients admitted with pneumonia . The course in one was complicated by overwhelming multiorgan failure when the only indicative laboratory result was seropositivity for Legionella hackeliae and Legionella longbeachae . He was initially treated with ceftriaxone and roxithromycin, followed by levofloxacin as well as intensive supportive treatment, and survived . The second patient was seroreactive for Legionella micdadei . In some cases of pneumonia in splenectomized patients tentatively considered to be caused by Streptococcus pneumoniae, the causative agent might have, in fact, been Legionella . We suggest that splenectomy be considered a possible predisposing factor for Legionella pneumonia . Since prompt diagnosis of Legionella infection, especially the non- pneumophila species, is extremely difficult, alertness to this diagnostic option and early empirical initiation of appropriate aggressive antibiotic treatment may be of critical importance . Copyright Urban and Vogel

Retina, 2004 Jun, 24(3), 391 - 8
Applicability of rapid antibiotic susceptibility testing in the management of bacterial endophthalmitis; Gupta R et al.; PURPOSE: To compare the applicability of the Kirby-Bauer disk diffusion method (conventional method) and the rapid antibiotic susceptibility test (RAST) for susceptibility testing of bacteria in the management of bacterial endophthalmitis . METHODS: Vitreous samples from 114 consecutive patients with endophthalmitis who were seen between June 2002 and October 2002 were subjected to microbiological processing (smears and cultures for bacteria and fungi) . Forty-seven of 114 samples could be tested by RAST . In addition, the bacterial isolates from 47 patients were tested for antibiotic susceptibility by the conventional method, and minimum inhibitory concentrations (MICs) of various antibiotics were determined . Clinical and microbiological data for these 47 patients were analyzed . RESULTS: Bacteria were isolated by routine culture from 18 (38.3%) of 47 samples; all 18 were tested by the conventional method, and MICs were determined for antibiotic susceptibility . Only 10 (55.6%) of these 18 samples were analyzed by RAST . Low inoculum and the slow-growing nature of the organisms (Streptococcus, 3; Staphylococcus epidermidis, 5) accounted for no growth on RAST plates in eight cases . Of four patients who were treated with repeated intraocular antibiotics, only one received RAST result-guided intraocular injection . Considering MIC testing as the gold standard, the sensitivity and specificity of RAST were either equal to (gentamicin, cefazolin, vancomycin, and chloramphenicol) or lower than (ciprofloxacin) those of the conventional technique . CONCLUSION: This comparative study failed to demonstrate a distinct advantage of RAST as a routine procedure in the management of bacterial endophthalmitis because of the need for trained staff to perform RAST, the nearly 50% negativity of culture, and the occasionally unreliable results.

J Immunol, 2004 Jun 15, 172(12), 7603 - 9
IL-10 is an important mediator of the enhanced susceptibility to pneumococcal pneumonia after influenza infection; van der Sluijs KF et al.; Secondary pneumococcal pneumonia is a serious complication during and shortly after influenza infection . We established a mouse model to study postinfluenza pneumococcal pneumonia and evaluated the role of IL-10 in host defense against Streptococcus pneumoniae after recovery from influenza infection . C57BL/6 mice were intranasally inoculated with 10 median tissue culture infective doses of influenza A (A/PR/8/34) or PBS (control) on day 0 . By day 14 mice had regained their normal body weight and had cleared influenza virus from the lungs, as determined by real-time quantitative PCR . On day 14 after viral infection, mice received 10(4) CFU of S . pneumoniae (serotype 3) intranasally . Mice recovered from influenza infection were highly susceptible to subsequent pneumococcal pneumonia, as reflected by a 100% lethality on day 3 after bacterial infection, whereas control mice showed 17% lethality on day 3 and 83% lethality on day 6 after pneumococcal infection . Furthermore, 1000-fold higher bacterial counts at 48 h after infection with S . pneumoniae and, particularly, 50-fold higher pulmonary levels of IL-10 were observed in influenza-recovered mice than in control mice . Treatment with an anti-IL-10 mAb 1 h before bacterial inoculation resulted in reduced bacterial outgrowth and markedly reduced lethality during secondary bacterial pneumonia compared with those in IgG1 control mice . In conclusion, mild self-limiting influenza A infection renders normal immunocompetent mice highly susceptible to pneumococcal pneumonia . This increased susceptibility to secondary bacterial pneumonia is at least in part caused by excessive IL-10 production and reduced neutrophil function in the lungs.

Obes Surg, 2004 May, 14(5), 690 - 4
Purpura fulminans due to Streptococcus pneumoniae sepsis following gastric bypass; Cone LA et al.; An older female underwent bariatric surgery which was followed by a significant weight loss and diarrhea, from which C . difficile was isolated just before her hospitalization . Less than 48 hours after admission, she became febrile, developed deep venous thrombosis of the leg and a pulmonary embolus . Blood cultures grew out Streptococcus pneumoniae and the patient developed purpura fulminans . There was convincing laboratory evidence for disseminated intravascular coagulation and a marked depletion of proteins C and S as well as antithrombin . Treatment with ceftriaxone and drotrecogin alfa together with parenteral nutrition led to disappearance of the pathogen and ultimate normalization of the anticoagulant factors . We believe that malabsorption of vitamin K dependent proteins C, S and antithrombin due to bariatric surgery predisposed the patient to purpura fulminans and disseminated intravascular coagulation.

J Appl Microbiol, 2004, 97(1), 149 - 57
Localization of MtuA, an LraI homologue in Streptococcus uberis; Jones CL et al.; AIMS: To determine the localization of MtuA, an LraI lipoprotein within Streptococcus uberis and assess whether the protein was able to induce an antibody response capable of growth inhibition . METHODS AND RESULTS: Immunoblots and ELISAs were performed on S . uberis cell fractions to localize the protein . The strongest reactivity was within the membrane-enriched fraction . Electron micrographs also showed labelling consistent with a location within the membrane . Specific antibodies from both rabbits and calves were unable to inhibit the growth of S . uberis in milk . In addition, MtuA was not detectable in a whole-cell ELISA and whole bacterial cells were unable to adsorb specific antibodies from antiserum raised against MtuA . CONCLUSIONS: The MtuA protein appears to be located within the cell membrane and is not on the bacterial surface and thus not available for interaction with potentially growth-inhibiting antibodies . SIGNIFICANCE AND IMPACT OF THE STUDY: Unlike PsaA of S . pneumoniae and MtsA of S . pyogenes, MtuA of S . uberis does not appear to be located at the cell surface . Therefore, in contrast to studies with other similar proteins, MtuA is unlikely to be a good vaccine candidate.

Mol Microbiol, 2004 Jun, 52(6), 1665 - 76
Specificity of streptolysin O in cytolysin-mediated translocation; Meehl MA et al.; Cytolysin-mediated translocation (CMT) is a recently described process in the Gram-positive pathogen Streptococcus pyogenes that translocates an effector protein of streptococcal origin into the cytoplasm of a host cell . At least two proteins participate in CMT, the pore-forming molecule streptolysin O (SLO) and an effector protein with the characteristics of a signal transduction protein, the Streptococcus pyogenes NAD-glycohydrolase (SPN) . In order to begin to elucidate the molecular details of the translocation process, we examined whether perfringolysin O (PFO), a pore-forming protein related to SLO, could substitute for SLO in the translocation of SPN . When expressed by S . pyogenes, PFO, like SLO, had the ability to form functional pores in keratinocyte membranes . However, unlike SLO, PFO was not competent for translocation of SPN across the host cell membrane . Thus, pore formation by itself was not sufficient to promote CMT, suggesting that an additional feature of SLO was required . This conclusion was supported by the construction of a series of mutations in SLO that uncoupled pore formation and competence for CMT . These mutations defined a domain in SLO that was dispensable for pore formation, but was essential for CMT . However, introduction of this domain into PFO did not render PFO competent for CMT, implying that an additional domain of SLO is also critical for translocation . Taken together, these data indicate that SLO plays an active role in the translocation process that extends beyond that of a passive pore.

Cell Microbiol, 2004 Jul, 6(7), 609 - 23
Whole-body autoradiography reveals that the Peptostreptococcus magnus immunoglobulin-binding domains of protein L preferentially target B lymphocytes in the spleen and lymph nodes in vivo; Smith D et al.; Protein L is an immunoglobulin (Ig)-binding protein produced by the Gram-positive bacterium Peptostreptococcus magnus that interacts with the variable region of Ig kappa light chains . The Ig light chain-binding capacity of protein L gives it the potential to interact with cells expressing surface Ig such as B cells . The present study was performed to address the in vivo trafficking of protein L at both the organ and the cellular level . Using the powerful technique of whole-body autoradiography in a murine model system, we demonstrate specific targeting of protein L to secondary lymphoid tissues in whole-animal analysis . The observed targeting depends on the capacity to interact with murine Ig, as tissue targeting was not apparent in mice given protein H, an Ig-binding protein produced by Streptococcus pyogenes with affinity for human but not murine Ig . Tissue targeting data were combined with flow cytometry analysis, which demonstrated the capacity of protein L to target and activate B lymphocytes in vivo . B cells targeted by protein L had increased surface expression of CD86 and MHC-II, and protein L was present in vacuolar compartments of B cells . Protein L did not bind T cells or natural killer cells but had some capacity to target dendritic cells and macrophages . The data show that protein L preferentially targets secondary lymphoid organs, and activates and is internalized by B cells in vivo . Furthermore, the observed tissue and cell targeting properties require an affinity for murine Ig . These data support the potential use of this Ig-binding protein as a targeting approach to deliver agents to defined cell populations in vivo.

Mil Med, 2004 May, 169(5), 373 - 5
A case series of group A Streptococcus necrotizing fasciitis in military trainees; Crum NF et al.; We describe a case series of group A Streptococcus (GAS) necrotizing fasciitis occurring over a 10-week period in military recruits undergoing intense physical training . These cases highlight that GAS may cause life-threatening infections in addition to mild diseases such as pharyngitis . This series suggests that the incidence of severe GAS infections may be increasing in certain populations and it emphasizes the importance of considering prophylactic measures against GAS in high-risk populations.

Schweiz Rundsch Med Prax, 2004 May 5, 93(19), 819 - 22
{Fever, malaise and new onset mitral valve insufficiency . Subacute Streptococcus bovis mitral valve endocarditis }; Burri E et al.; A 62-year-old patient with low grade fever, fatigue, arthralgia and newly discovered mitral regurgitation was diagnosed with subacute endocarditis . Streptococcus bovis grew from all six blood culture bottles . Streptococcus bovis is known to be associated with gastrointestinal neoplasias . Therefore a colonoscopy was performed and two polyps were removed . Histological analysis revealed a tubulovillous adenoma and a serrated adenoma . Colonoscopy is mandatory for all patients with Streptococcus bovis endocarditis even without any symptoms for colorectal neoplasia . The significance of Streptococcus bovis for the carcinogenesis of colorectal neoplasias and the possible alternative pathway for colorectal carcinomas through serrated adenomas will be discussed.

Schweiz Monatsschr Zahnmed, 2004, 114(4), 337 - 41
{Bacterial contamination of bony particles from the bone collection trap}; Glaser B et al.; Intraoral bony defects can be filled with bony particles that are collected in a titanium filter while drilling . The rinsing liquid is contaminated with blood and saliva which implies that the bony particles are also contaminated with bacteria . The aim of this study was to determine quantitatively and qualitatively the degree of this contamination . Over a period of three months bony particles were collected from 50 patients undergoing surgery . The bony particles were scraped off the filter, resuspended and incubated aerobically and anaerobically on human blood agar media . Colony forming units (CFU) were determined as well as the most common species of bacteria . All samples showed anaerobic and aerobic growth . After anaerobic incubation in 44 samples the number of bacteria was higher (38) or equal (six) to that after aerobic incubation . On average 435,000 CFU (aerobic) and 1,013,000 CFU (anaerobic) per sample were found . The most frequently identified bacteria belonged to Veillonella spp . in the anaerobic and to Streptococcus oralis in the aerobic cultures . In 43 samples black pigmented colonies were detected . There were only bacteria identified which are common in the oral cavity.

J Med Microbiol, 2004 Jul, 53(Pt 7), 697 - 703
Genotypic diversity and virulence traits of Streptococcus mutans in caries-free and caries-active individuals; Napimoga MH et al.; The present study evaluated the relationship between clonal diversity and some virulence traits of Streptococcus mutans isolated from eight caries-free and eight caries-active subjects . A total of 155 S . mutans isolates from caries-free subjects and 144 isolates from caries-active subjects were obtained from samples of saliva, dental plaque and tongue surface and identified by PCR . The isolates were submitted to arbitrarily primed (AP)-PCR (OPA-2 and OPA-13) and multilocus enzyme electrophoresis (MLEE) to establish the genotypic diversity . Production of water-insoluble glucan (WIG) (monitored by SDS-PAGE), final pH of cultures and the ability of bacterial cells to adhere to smooth glass in the presence of sucrose were measured . High and comparable abilities of MLEE and AP-PCR were found to distinguish S . mutans genotypes, using Simpson's index of discrimination (0.971 and 0.968, respectively) . The results showed a significant difference (P < 0.01) in the number of genotypes when caries-free and caries-active groups were compared by both fingerprinting methods used . Final pH (P = 0.32) and the percentage of adherence to a glass surface (P = 0.62) did not show differences between the two groups; however, the intensities of WIG bands from the caries-active group were greater than those from the caries-free group (P < 0.01) . In addition, WIG was positively correlated with the ability of S . mutans to adhere to a glass surface (r = 0.34, P = 0.02) from caries-active subjects . These data showed that AP-PCR analysis and MLEE are both effective methods for assessing the genetic relatedness of S . mutans . Using these techniques, it was found that there is a larger number of genotypes of S . mutans with increased ability to synthesize WIG in caries-active individuals.

J Med Microbiol, 2004 Jul, 53(Pt 7), 645 - 51
Aetiology of acute pharyngitis: the role of atypical bacteria; Esposito S et al.; In order to establish the role of atypical bacteria and compare characteristics of different infectious agents in acute pharyngitis, 127 patients with acute pharyngitis (66 males; median age, 5.33 years; range, 6 months to 14 years) and 130 healthy subjects of similar sex and age were studied . Serology with paired samples and PCR on nasopharyngeal aspirates and throat cultures were used to identify bacteria and viruses . Viruses were identified in 43 patients (33.8%) and five controls (3.8%; P < 0.0001), potential bacterial pathogens in 34 patients (26.8%) and 26 controls (20%; P = 0.256) and mixed viral/bacterial pathogens in 26 patients (20.5%) and none of the controls (P < 0.0001) . The main aetiological agents were adenovirus, respiratory syncytial virus (RSV), Mycoplasma pneumoniae, Streptococcus pyogenes and Chlamydia pneumoniae . M . pneumoniae was the agent found most frequently as a single pathogen . A history of recurrent pharyngitis, having older siblings and a negative outcome were significantly more common among patients with acute M . pneumoniae infection than among those with infections due to other pathogens or healthy controls . This study demonstrates that: (i) adenovirus and RSV have a prominent role in acute pharyngitis; (ii) S . pyogenes is found frequently, but it is not possible to distinguish simple carriers from patients with a true infection; (iii) M . pneumoniae appears to be able to cause acute pharyngitis per se; and (iv) C . pneumoniae seems to be mainly a co-pathogen . To avoid the risk of an incorrect therapeutic approach, simple laboratory investigations that allow rapid identification of M . pneumoniae infections are urgently needed.

J Med Microbiol, 2004 Jul, 53(Pt 7), 595 - 602
PCR-based assays for detection of Streptococcus pneumoniae serotypes 3, 14, 19F and 23F in respiratory specimens; Rubin LG et al.; Current culture-based assays are insensitive for detection of simultaneous respiratory tract colonization by more than one pneumococcal serotype . Separate single-tube, nested PCR-based assays have been developed to detect Streptococcus pneumoniae serotypes 3, 14, 19F and 23F by amplifying unique DNA sequences in the capsular polysaccharide gene cluster of each serotype . Pairs of 27-32-base outer primers and 20-21-base inner primers and a 20-22-base probe were designed to amplify and detect a 200-221-base sequence by dot blotting using the labelled probe . Sensitivity of the assays was 0.01-10 fg using chromosomal DNA and < or = 1 viable cell using DNA extracted from exponential-phase bacteria . Each serotype-specific assay detected chromosomal DNA from all of five to ten clinical isolates of the homologous type and did not detect DNA sequences from any of 190-204 strains from 51-52 different serotypes or 28 non-pneumococcal bacterial strains . Sixteen throat swabs from children that had been cultured for S . pneumoniae were tested in PCR assays following DNA extraction . All of six that grew S . pneumoniae serotype 3, 14, 19F or 23F were positive in the PCR assay for the homologous serotype (and in a PCR assay for sequences in lytA, present in all pneumococci) and were negative in assays for other serotypes . Of eight culture-negative specimens in children not receiving antimicrobials, three were positive for both the lytA assay and an assay for one of the four serotypes, suggesting true positive results; in three others all five PCR assays were negative and, in the remaining two, the lytA assay was positive but each of the four assays for individual serotypes was negative, suggesting either false-positive results or presence of DNA sequences from an S . pneumoniae serotype other than 3, 14, 19F or 23F . These preliminary clinical data suggest that these PCR-based assays are sensitive and specific for detection of individual serotypes of pneumococci and may be used with respiratory tract specimens.

J Clin Microbiol, 2004 Jun, 42(6), 2810 - 2
Clonality of Streptococcus pneumoniae serotype 1 isolates from pediatric patients in the United States; Gonzalez BE et al.; We compared Streptococcus pneumoniae serotype 1 isolates causing disease among children in six geographic regions of the United States to determine genetic relatedness . Genomic fingerprints were determined by repetitive element polymorphism PCR (Rep-PCR) . Multilocus sequence type characterization was performed on selected isolates . Four different genomic banding patterns were identified by Rep-PCR . One profile (clone 1) was predominant and matched sequence type 227.

J Clin Microbiol, 2004 Jun, 42(6), 2518 - 22
Simple, rapid latex agglutination test for serotyping of pneumococci (Pneumotest-Latex); Slotved HC et al.; The "gold standard" for epidemiological typing of Streptococcus pneumoniae (pneumococcus) is the capsular reaction test (Neufeld test) with antisera against the 90 pneumococcal polysaccharide capsules, i.e., serotyping . The method is labor intensive and requires a certain level of experience to be performed satisfactory, and thus it has been restricted for use in specialized reference or research laboratories . Surveillance of the serotype distribution of pneumococci that cause infections is important to secure an optimal composition of pneumococcal vaccines and to monitor antibiotic resistance in pneumococci . At Statens Serum Institut, a simple latex agglutination test for serotyping of pneumococci has been developed . The Pneumotest-Latex kit consists of 14 different pooled pneumococcus antisera (pools A to I and pools P to T) applied to latex particles . In a blind test of 352 isolates (with all 90 serotypes represented), 336 (95.5%) were typed or grouped correctly by the Pneumotest-Latex; in addition, 2 (7%) of 30 strains regarded as nontypeable or rough strains were serotyped, and the serotypes of these two isolates were confirmed by the capsular reaction test with type-specific antisera . The Pneumotest-Latex seems to be a sensitive method for serotyping or grouping of the majority of pneumococcal strains . By use of this ready-to-perform latex agglutination kit (Pneumotest-Latex), serotyping of pneumococci can gain more ground as a tool in prevention of pneumococcal diseases.

Am J Respir Crit Care Med, 2004 Aug 15, 170(4), 440 - 4 Epub 2004 Jun 07.
Combination antibiotic therapy lowers mortality among severely ill patients with pneumococcal bacteremia; Baddour LM et al.; Retrospective studies have suggested that combination antibiotic therapy for severe bacteremic pneumococcal pneumonia may reduce mortality . We assessed this issue in a prospective, multicenter, international observational study of 844 adult patients with bacteremia due to Streptococcus pneumoniae . The effect of combination antibiotic therapy versus monotherapy on mortality was examined by univariate analyses and by logistic regression models . The 14-day mortality was not significantly different for the two groups . However, among critically ill patients, combination antibiotic therapy was associated with lower 14-day mortality (23.4 versus 55.3%, p = 0.0015) . This improvement in survival was independent of country of origin, intensive care unit support, class of antibiotics, or in vitro activity of the antibiotics prescribed . Combination antibiotic therapy improved survival among critically ill patients with bacteremic pneumococcal illness.

Diagn Microbiol Infect Dis, 2004 Jun, 49(2), 147 - 9
Doxycycline use for community-acquired pneumonia: contemporary in vitro spectrum of activity against Streptococcus pneumoniae (1999-2002); Jones RN et al.; Because of the limited clinical testing of doxycycline, it has received guarded acceptance as an inexpensive, fluoroquinolone-sparing agent for treatment of community-acquired pneumonia . Examination of in vitro data from 3,902 recent Streptococcus pneumoniae isolates (SENTRY Program, USA) suggests that doxycycline has wider clinical application compared to macrolides, oral cephalosporins (directed by penicillin susceptibility), and trimethoprim-sulfamethoxazole .

Lancet, 2004 Jun 5, 363(9424), 1871 - 2
Colonisation by Streptococcus pneumoniae and Staphylococcus aureus in healthy children; Bogaert D et al.; A trial with a 7-valent pneumococcal-conjugate vaccine in children with recurrent acute otitis media showed a shift in pneumococcal colonisation towards non-vaccine serotypes and an increase in Staphylococcus aureus-related acute otitis media after vaccination . We investigated prevalence and determinants of nasopharyngeal carriage of Streptococcus pneumoniae and S aureus in 3198 healthy children aged 1-19 years . Nasopharyngeal carriage of S pneumoniae was detected in 598 (19%) children, and was affected by age (peak incidence at 3 years) and day-care attendance (odds ratio {OR} 2.14, 95% CI 1.44-3.18) . S aureus carriage was affected by age (peak incidence at 10 years) and male sex (OR 1.46, 1.25-1.70) . Serotyping showed 42% vaccine type pneumococci . We noted a negative correlation for co-colonisation of S aureus and vaccine-type pneumococci (OR 0.68, 0.48-0.94), but not for S aureus and non-vaccine serotypes . These findings suggest a natural competition between colonisation with vaccine-type pneumococci and S aureus, which might explain the increase in S aureus-related otitis media after vaccination.

Zhonghua Nei Ke Za Zhi, 2004 May, 43(5), 329 - 32
{The phenotype and genotype patterns of erythromycin-resistant Streptococcus pneumoniae}; Zhao TM et al.; OBJECTIVE: To investigate the resistance phenotypes and genotypes in erythromycin-resistant Streptococcus (S.) pneumoniae . METHODS: The minimum inhibitory concentration (MIC) of erythromycin, clindamycin, penicillin and fluoroquinolones against 192 strains of S . pneumoniae was tested with broth microdilution method according to the guidelines of the National Committee for Clinical Laboratory Standards . Of 148 clinical isolates of erythromycin resistant S . pneumoniae, the macrolide resistance phenotypes were observed by the erythromycin-clindamycin-spiramycin triple-disc test and erythromycin resistance genes were detected by polymerase chain reaction . RESULTS: 42.7% S . pneumoniae isolates was resistant (intermediate and resistant) to penicillin while the resistance rates to erythromycin and clindamycin were 77.6% and 66.7% respectively . The ermB gene, being the most prevalent, was detected in 79.1% of the 148 erythromycin-resistant strains . The main phenotype (85.1%) of erythromycin-resistant strains was constitutive macrolide, lincosamide, and streptogramin B resistance phenotype (cMLS) . Erythromycin MICs for S . pneumoniae ermB-positive isolates were higher than those for mefA-positive isolates . 74.4% of the ermB-positive isolates demonstrated erythromycin MICs of > 16.0 micro g/ml, and the erythromycin MICs for mefA-positive isolates ranged from 0.5 approximately 4.0 micro g/ml . CONCLUSIONS: The resistance rate of S . pneumoniae to erythromycin is high in China . The main phenotype is cMLS . Ribosomal modification (ermB gene coded) is the main resistance mechanism against erythromycin in S . pneumoniae

Treat Respir Med, 2004, 3(2), 67 - 77
Guidelines for community-acquired pneumonia: are they reflected in practice?
Flanders SA, Halm EA.
Community-acquired pneumonia (CAP) is common, costly, and clinically serious . Several national and international practice guidelines have been developed to promote more appropriate, cost-effective care for patients with CAP . This article compares and contrasts eight international practice guidelines for the management of CAP, describes the extent to which recommendations are reflected in practice, and proposes explanations for non-adherence to guidelines . We found consistency in recommendations across all the guidelines for the management of patients with CAP requiring intensive care . In this setting, all guidelines recommend chest radiography, sputum Gram stain and culture, blood cultures, testing for Legionella pneumophila, and timely administration of antibiotics active against both typical (i.e . Streptococcus pneumoniae, Hemophilus influenzae) and atypical organisms (i.e . Legionella spp., Mycoplasma pneumoniae, and Chlamydia pneumoniae) . Recommendations for the management of the average inpatient with pneumonia were more variable, with the greatest differences between the North American and European guidelines . The North American guidelines (in contrast to European ones), recommended empiric treatment of typical and atypical organisms in all inpatients . There were also differences in policies regarding the necessity of chest radiography, sputum studies, and serologic testing . Some guidelines explicitly embrace the use of prediction rules to inform the decision to hospitalize, while others do not . Some of these admission decision algorithms focus on identifying low risk patients, while others are most concerned with high risk patients . There was also considerable variation in the specificity and operationalization of clinical criteria for switching from parenteral to oral antibiotics or judging appropriateness for discharge . Many recommendations for key management decisions tended to lack explicit, objective, and actionable criteria that could be easily implemented in real world practice . Review of the pneumonia literature revealed that physician performance of guideline-recommended best practices is often suboptimal . Administration of timely antibiotics (< or =8 hours of presentation) and use of first-line antibiotics occurred in 75-85% and 18-79% of cases, respectively . Collection of blood cultures within 24 hours of presentation and prior to administration of antibiotics was achieved in 69-83% and 63-82% of cases, respectively . Screening the eligibility of CAP patients for hospital-based pneumococcal and influenza vaccination occurred on average in 11 and 14% of hospitalizations, respectively, in the US . Lack of awareness of guidelines, conflicting advice among them, and lack of specific, objective, actionable recommendations most likely contribute to nonadherence to CAP guidelines . Increased attention to these factors will be needed if professional society practice guidelines are to fulfill their promise as tools for improving the quality and outcomes of care for patients with pneumonia.

Cutis, 2004 May, 73(5 Suppl), 14 - 8
Overview of cefdinir: pharmacokinetics, safety, and efficacy in the treatment of uncomplicated skin and skin structure infections; Paris MM et al.; Uncomplicated skin and skin structure infections commonly are caused by Staphylococcus aureus and Streptococcus pyogenes . Cefdinir, an extended spectrum, third-generation cephalosporin is a safe and effective means of treating skin infections caused by these organisms, as well as many gram-negative pathogens . This article summarizes the pharmacokinetics, dosing schedule, adverse event profile, and efficacy data for cefdinir in adult and pediatric, populations in the treatment of uncomplicated skin and skin structure infections.

Cutis, 2004 May, 73(5 Suppl), 7 - 10
Common pathogens and differential diagnosis of skin and soft tissue infections; Murakawa GJ; The major organisms responsible for skin infections are Staphylococcus aureus and Streptococcus pyogenes . To produce a disease state, both organisms must overcome the body's natural defenses, adhere to the skin, invade the tissue, and proliferate . Often, endogenous skin microflora can lead to systemic infections, especially in immunocompromised individuals . The differential diagnosis of skin infections is key to successful therapy and requires a thorough knowledge of the patient's clinical history and immune status . Laboratory analysis often can assist in the pathogenic differentiation . Examples of common skin infections are illustrated herein.

Laryngoscope, 2004 Jun, 114(6), 1032 - 6
Cathepsin gene expression profile in rat acute pneumococcal otitis media; Li-Korotky HS et al.; OBJECTIVES/HYPOTHESIS: Acute otitis media, often caused by infection with Streptococcus pneumoniae, is characterized by inflammation of the middle ear mucosa . A prominent feature of the host response to bacterial infection of the middle ear mucosa is an influx of inflammatory cells that contributes to the local pool of inflammatory mediators by releasing additional inflammatory chemicals, which in turn cause further tissue injury . The objective was to identify candidate effector and signaling molecules involved in acute otitis media pathogenesis caused by S pneumoniae infection . STUDY DESIGN: Male Sprague-Dawley rats were randomly assigned to 1 of 5 groups, including 1 control group without treatment, 2 placebo groups (12 and 48 hours) and 2 infected groups (12 and 48 hours) . The rat middle ear was bilaterally inoculated with either 25 microL of tryptic soy broth (TSB group) or 25 microL of TSB containing approximately 1.24 x 10(9) cfu/mL of S pneumonias type 6A (SP group) . Rats were killed at 12 and 48 hours after inoculation and the middle ear mucosa was collected . Total RNA was extracted and pooled from each group for gene expression assays . METHODS: Gene expression profiles for rat middle ear mucosa at 12 and 48 hours after S pneumoniae or placebo inoculation were constructed using microarray technology (Clontech Atlas Rat 1.2 Array, 1176 cDNAs) . Genes of interest were further validated by real-time polymerize chain reaction . RESULTS: Middle ear mucosa expression of a gene cluster encoding the lysosomal cysteine proteases, cathepsins B (Ctsb), L (Ctsl), and K (Ctsk), was modified after S pneumoniae challenge . Specifically, at 12 hours, Ctsk and Ctsl messenger RNA that was abundantly expressed in the normal middle ear mucosa was decreased, whereas Ctsb transcript was induced . The changes in Ctsb and Ctsk gene expression were sustained at 48 hours . CONCLUSION: The constitutive expression of Ctsk and Ctsl messenger RNA in normal middle ear mucosa supports a function in the maintenance of middle ear mucosa homeostasis, and their downregulation as an early event in acute otitis media may reflect a disruption in that function . The induction of Ctsb messenger RNA in the infected middle ear mucosa suggests a role in early tissue injury; thus, Ctsb may represent a potential target for molecular diagnostics and/or rational intervention during the development of acute otitis media.

Laryngoscope, 2004 Jun, 114(6), 975 - 80
Impact of pneumococcal polysaccharide vaccine (Prevnar) on middle ear fluid in children undergoing tympanostomy tube insertion; Caspary H et al.; OBJECTIVES: The purpose of this study was to describe the bacteriology of middle ear effusions in children who have received the pneumococcal polysaccharide vaccine (Prevnar) compared with unvaccinated children . METHODS: A prospective review of medical records from July 2001 to July 2002 was conducted on children with middle ear effusion at time of tympanostomy tube insertion . Middle ear fluid was plated onto culture media immediately after acquisition, and antimicrobial resistance of cultured organisms along with serotyping of Streptococcus pneumoniae was examined . Vaccination status, demographics, and risk factors were determined from patients' medical records, parent interviews, or contact with their primary care physicians . RESULTS: After adjusting for age and number of previous infections, children vaccinated with Prevnar are two times less likely to have non-S . pneumoniae pathogenic bacteria isolated than children not vaccinated . Of those with growth, vaccinated children were almost three times more likely than nonvaccinated children to have the presence of H . influenzae . Vaccinated children with H . influenzae were 7.5 times less likely to have beta-lactamase producing H . influenzae than nonvaccinated children with H . influenzae . CONCLUSION: Because the incidence of S . pneumoniae was low, no inference could be made whether Prevnar decreased otitis media with effusion or recurrent acute otitis caused by the S . pneumoniae serotypes covered by the vaccine . However, vaccinated children did appear to have the unexpected benefit of having a certain level of protection to growth of typical acute otitis media pathogens.

Science, 2004 Jun 4, 304(5676), 1513 - 5
A microdomain for protein secretion in Gram-positive bacteria; Rosch J et al.; Gram-positive bacteria face unique challenges in generating biologically active conformations for their exported proteins because they lack a dedicated compartment for folding secreted polypeptides . We have discovered that protein secretion by way of the general secretory (Sec) pathway in the important human pathogen Streptococcus pyogenes proceeds through a single microdomain . Unlike other mechanisms for asymmetry involving the Sec pathway, proteins destined for secretion are targeted to a single locus distal to either cell pole that has specialized to contain the Sec translocons . This subcellular organization may represent a paradigm for secretion common to other Gram-positive pathogens with profound implications for pathogenesis.

Biochem Biophys Res Commun, 2004 Jun 25, 319(2), 439 - 47
Characterisation of a novel homodimeric N-acetyl-beta-D-glucosaminidase from Streptococcus gordonii; Harty DW et al.; An N-acetyl-beta-D-glucosaminidase (GcnA) from Streptococcus gordonii FSS2 was cloned and sequenced . GcnA had a deduced molecular mass of 72,120 Da . The molecular weight after gel-filtration chromatography was 140,000 Da and by SDS-PAGE was 70,000 Da, indicating that the native protein was a homodimer . The deduced amino acid sequence had significant homology to a glycosyl hydrolase from Streptococcus pneumoniae and the conserved catalytic domain of the Family 20 glycosyl hydrolases . GcnA catalysed the hydrolysis of the synthetic substrates, 4-methylumbelliferyl (4MU)-N-acetyl-beta-D-glucosaminide, 4MU-N-acetyl-beta-D-galactosaminide, 4-MU-beta-D-N,N'-diacetylchitobioside, and 4-MU-beta-D-N,N',N''-chitotrioside as well as the respective chito-oligosaccharides . GcnA was optimally active at pH 6.6 and 42 degrees C . The Km for 4-MU-beta-D-N,N',N''-chitotrioside, 45 microM, was the lowest for all the substrates tested . Hg2+, Cu2+, Fe2+, and Zn2+ completely inhibited while Co2+, Mn2+, and Ni2+ partially inhibited activity . S . gordonii FSS2 and a GcnA negative mutant grew equally well on chito-oligosaccharides as substrates . The S . gordonii sequencing projects indicate two further N-acetyl-beta-D-glucosaminidase activities.

Protein Expr Purif, 2004 Jul, 36(1), 82 - 9
Cloning, overexpression, and characterization of a serine/threonine protein kinase pknI from Mycobacterium tuberculosis H37Rv; Gopalaswamy R et al.; Protein phosphorylation-dephosphorylation is the principal mechanism for translation of external signals into cellular responses . Eukaryotic-like serine/threonine kinases have been reported to play important roles in bacterial development and/or virulence . The PknI protein is one of the 11 eukaryotic-like serine/threonine kinases in Mycobacterium tuberculosis H37Rv . From the bioinformatic studies, PknI protein has been shown to have an N-terminal cytoplasmic domain followed by a transmembrane region and an extracellular C-terminus suggestive of a sensor molecule . In this study, we have cloned, overexpressed, and characterized the entire coding region and the cytoplasmic domain of PknI as a fusion protein with an N-terminal histidine tag, and used immobilized metal affinity chromatography for purification of recombinant proteins . The purified recombinant proteins were found to be functionally active through in vitro phosphorylation assay and phosphoamino acid analysis . In vitro kinase assay of both proteins revealed that PknI is capable of autophosphorylation and showed manganese-dependent activity . Phosphoamino acid analysis indicated phosphorylation at serine and threonine residues . Southern blot analysis with genomic DNA highlighted the conserved nature of pknI among the various mycobacterial species . In silico analysis revealed a close homology of PknI to Stk1 from Streptococcus agalactiae, shown to have a role in virulence and cell segregation of the organism.

J Bacteriol, 2004 Jun, 186(12), 3928 - 37
CovS inactivates CovR and is required for growth under conditions of general stress in Streptococcus pyogenes; Dalton TL et al.; The gram-positive human pathogen Streptococcus pyogenes (group A streptococcus {GAS}) causes diseases ranging from mild and often self-limiting infections of the skin or throat to invasive and life-threatening illnesses . To cause such diverse types of disease, the GAS must be able to sense adverse environments and regulate its gene expression accordingly . The CovR/S two-component signal transduction regulatory system in GAS represses about 15% of the GAS genome, including many genes involved in virulence, in response to the environment . We report that CovR is still able to repress transcription from several promoters in the absence of the putative histidine kinase sensor for this system, CovS . We also show that a phosphorylation site mutant (D53A) of CovR is unable to repress gene expression . In addition, we report that a strain with a nonpolar mutation in CovS does not grow at a low pH, elevated temperature, or high osmolarity . The stress-related phenotypes of the CovS mutant were complemented by expression of covS from a plasmid . Selection for growth of a CovS mutant under stress conditions resulted in isolation of second-site mutations that inactivated covR, indicating that CovR and CovS act in the same pathway . Also, at 40 degrees C in the wild-type strain, CovR appeared to be less active on the promoter tested, which is consistent with the hypothesis that it was partially inactivated by CovS . We suggest that under mild stress conditions, CovS inactivates CovR, either directly or indirectly, and that this inactivation relieves repression of many GAS genes, including the genes needed for growth of GAS under stress conditions and some genes that are necessary for virulence . Growth of many gram-positive bacteria under multiple-stress conditions requires alteration of promoter recognition produced by RNA polymerase association with the general stress response sigma factor, sigma(B) . We provide evidence that for GAS, which lacks a sigB ortholog, growth under stress conditions requires the CovR/S two-component regulatory system instead . This two-component system in GAS thus appears to perform a function for which other gram-positive bacteria utilize an alternative sigma factor.

J Bacteriol, 2004 Jun, 186(12), 3721 - 9
A homologue of aliB is found in the capsule region of nonencapsulated Streptococcus pneumoniae; Hathaway LJ et al.; The epidemiology, phylogeny, and biology of nonencapsulated Streptococcus pneumoniae are largely unknown . Increased colonization capacity and transformability are, however, intriguing features of these pneumococci and play an important role . Twenty-seven nonencapsulated pneumococci were identified in a nationwide collection of 1,980 nasopharyngeal samples and 215 blood samples obtained between 1998 and 2002 . On the basis of multilocus sequence typing and capsule region analysis we divided the nonencapsulated pneumococci into two groups . Group I was closely related to encapsulated strains . Group II had a clonal population structure, including two geographically widespread clones able to cause epidemic conjunctivitis and invasive diseases . Group II strains also carried a 1,959-bp homologue of aliB (aliB-like ORF 2) in the capsule region, which was highly homologous to a sequence in the capsule region of Streptococcus mitis . In addition, strains of the two major clones in group II had an additional sequence, aliB-like ORF 1 (1,968 to 2,004 bp), upstream of aliB-like ORF 2 . Expression of aliB-like ORF 1 was detected by reverse transcription-PCR, and the corresponding RNA was visualized by Northern blotting . A gene fragment homologous to capN of serotypes 33 and 37 suggests that group II strains were derived from encapsulated pneumococci some time ago . Therefore, loss of capsule expression in vivo was found to be associated with the importation of one or two aliB homologues in some nonencapsulated pneumococci.

Arch Soc Esp Oftalmol, 2004 May, 79(5), 213 - 19
{Preoperative eye-drop antibiotherapy in cataract surgery}; Fernandez Rubio E et al.; OBJECTIVE: To ascertain the effectiveness of various antibiotic eye-drops in eradicating the preoperative conjunctival bacteria of patients undergoing cataract surgery and to differentiate the failure of these treatments due to the lack of "in vitro" sensibility from other possible causes . METHODS: Retrospective study of the preoperative conjunctival flora of 4876 consecutive patients; "in vitro" sensibility was analysed by grouping bacteria into eight categories; the susceptibility percentages of the total conjunctival flora to five antibacterial agents were compared . The effectiveness of the eye-drop treatment with a single sensitive tested antibiotic (Aureomicin, Chloramphenicol, Gentamicin, Norfloxacin or Rifamicin) was evaluated in patients with pathogen bacteria . RESULTS: The "in vitro" sensibilities of Chloramphenicol (84.4%) and Rifampicin (83.9%) were similar (p < 0.01) and statistically higher than those of the other antibiotics . Nevertheless, the Chloramphenicol pathogen bacterium treatment failed in 21.2% of cases, in spite of being "in vitro" sensitive . Gentamicin presented the best effectiveness for eradicating Staphylococcus aureus and Gram (-) rods . Aureomicin had the best effectiveness against Streptococcus and Gram (-) diplococci . Rifamicin was the most effective for eradicating the whole predominant Gram (+) flora . The effectiveness of all five antibiotics decreased when there was more than one pathogen . CONCLUSIONS: None of the five antibiotic monotherapies maintains the patients' conjunctive free of pathogen bacteria 48 hours after finishing the treatment; however, there are bacterial patrons whose treatment could be optimised . The existence of polymicrobial flora decreases the effectiveness of the treatment.

Pediatrics, 2004 Jun, 113(6), 1735 - 40
The changing face of pleural empyemas in children: epidemiology and management; Schultz KD et al.; OBJECTIVE: Empyema remains a significant cause of morbidity in children . This study evaluates the changes that have affected the outcome in children with pleural empyema, including the emergence of resistant organisms, the introduction of the pneumococcal conjugate vaccine, and earlier treatment with video-assisted thoracoscopy (VATS) . METHODS: A retrospective chart review was performed on all patients who were discharged with a diagnosis of empyema and community-acquired pneumonia over a 10-year period (1993-2002) at Texas Children's Hospital in Houston, Texas . Data collected included demographic information, clinical presentation, radiographic studies, laboratory data including culture results, and hospital course . RESULTS: A total of 230 charts were available for review . The mean age of the patients was 4.0 +/- 3.6 years . Of the pleural fluid cultures performed, 32% (69 of 219) were positive . An additional 27 patients had a cause identified by blood culture . The first penicillin-nonsusceptible Streptococcus pneumoniae was identified in 1995, and the first methicillin-resistant Staphylococcus aureus was identified in 1998 . After the universal use of the pneumococcal conjugate vaccine, 3 major changes have occurred (1999-2000 vs 2001-2002): 1) the number of patients admitted with empyema (per 10 000 admissions) has decreased from 23 to 12.6; 2) the prevalence of S pneumoniae has decreased from 66% (29 of 44) to 27% (4 of 15); and 3) S aureus has become the most common pathogen isolated (18% vs 60%), with 78% of those being methicillin resistant . The use of early VATS (<48 hours after admission) versus late VATS (>48 hours after admission) significantly decreased the length of hospitalization (11.49 +/- 6.56 days vs 15.18 +/- 8.62 days) . CONCLUSIONS: The microbiologic cause of empyema has changed with an increasing incidence of S aureus, particularly methicillin-resistant S aureus . The use of VATS for initial therapy of empyema results in decreased duration of fever and length of hospitalization.

J Biol Chem, 2004 Aug 27, 279(35), 36250 - 8 Epub 2004 Jun 01.
Novel lipoglycopeptides as inhibitors of bacterial signal peptidase I; Kulanthaivel P et al.; Signal peptidase (SPase) I is responsible for the cleavage of signal peptides of many secreted proteins in bacteria . Because of its unique physiological and biochemical properties, it serves as a potential target for development of novel antibacterial agents . In this study, we report the production, isolation, and structure determination of a family of structurally related novel lipoglycopeptides from a Streptomyces sp . as inhibitors of SPase I . Detailed spectroscopic analyses, including MS and NMR, revealed that these lipoglycopeptides share a common 14-membered cyclic peptide core, an acyclic tripeptide chain, and a deoxy-alpha-mannose sugar, but differ in the degree of oxidation of the N-methylphenylglycine residue and the length and branching of the fatty acyl chain . Biochemical analysis demonstrated that these peptides are potent and competitive inhibitors of SPase I with K(i) 50 to 158 nm . In addition, they showed modest antibacterial activity against a panel of pathogenic Gram-positive and Gram-negative bacteria with minimal inhibitory concentration of 8-64 microm against Streptococcus pneumonniae and 4-8 microm against Escherichia coli . Notably, they mechanistically blocked the protein secretion in whole cells as demonstrated by inhibiting beta-lactamase release from Staphylococcus aureus . Taken together, the present discovery of a family of novel lipoglycopeptides as potent inhibitors of bacterial SPase I may lead to the development of a novel class of broad-spectrum antibiotics.

Vet Microbiol, 2004 Jun 21, 101(2), 117 - 22
Determination of species-specific sequences of superoxide dismutase A encoding gene sodA and chaperonin 60 encoding gene cpn60 for identification and phylogenetic analysis of Streptococcus phocae; Alber J et al.; Species-specific PCR tests, based on the manganese-dependent superoxide dismutase A encoding gene (sodA) and the chaperonin 60 encoding gene (cpn60), were developed for the identification of Streptococcus phocae, a bacterial pathogen of seals . The selection of both oligonucleotide primer pairs was performed after amplification and sequencing of internal parts of both genes using universal oligonucleotide primers . The sequence studies of both genes additionally confirmed that S . phocae could taxonomically be classified to the pyogenic group of the genus Streptococcus .

Vet Microbiol, 2004 Jun 21, 101(2), 109 - 16
Development of a PCR assay for Streptococcus iniae based on the lactate oxidase (lctO) gene with potential diagnostic value; Mata AI et al.; Streptococcus iniae is a well-known pathogen of both fish and humans that is difficult to identify by conventional biochemical tests . A PCR assay based on the lactate oxidase (lctO) gene of S . iniae was developed for the rapid and specific detection and identification of this pathogen from different sources . The PCR assay had a detection limit of 62-31 cells, and 25 pg of DNA per PCR reaction mixture . The PCR was also effective in detecting the bacterium from inoculated tissue homogenates, suggesting its potential use for a rapid and accurate diagnosis of S . iniae infections .

Medicina (Kaunas), 2004, 40(5), 414 - 8
{Poststreptococcal reactive arthritis}; Cerniauskiene V et al.; Arthritis, following infection caused by group A beta-hemolytic streptococcus, is classically attributed to acute rheumatic fever . However, a new clinical syndrome, called poststreptococcal reactive arthritis, as a distinct entity from acute rheumatic fever, was described recently . The purpose of this paper is to provide a summary of published information on poststreptococcal reactive arthritis . The paper outlines its clinical description and proposed diagnostic criteria . Similarities and differences between poststreptococcal reactive arthritis and acute rheumatic fever are discussed . Information regarding long-term risk of carditis following poststreptococcal reactive arthritis is provided, and therapeutic recommendations are outlined.

Am J Kidney Dis, 2004 Jun, 43(6), 976 - 82
Non-enteropathic hemolytic uremic syndrome: causes and short-term course; Constantinescu AR et al.; BACKGROUND: Nondiarrheal or Streptococcus pneumoniae-related hemolytic uremic syndrome (HUS) represents a heterogeneous group of disorders . This study was performed to: (1) describe the current incidence, causes, demographic features, hospital courses, and short-term outcomes of non-enteropathic HUS; (2) compare findings in patients with non-enteropathic HUS with those obtained from a contemporaneous cohort of children with enteropathic or diarrhea-associated HUS (D+ HUS) diagnosed and treated at the same clinical sites; and (3) identify clinical or laboratory features that differentiate these 2 groups and predict disease severity and the short-term outcome in patients with non-enteropathic HUS . METHODS: Data were collected from patients screened between 1997 and 2001 for enrollment in a multicenter trial of SYNSORB Pk (SYNSORB Biotech Inc, Calgary, Alberta, Canada) in D+ HUS, but who were ineligible because of lack of a diarrhea prodrome . The following features were recorded: age; sex; ethnicity; prodromal symptoms; cause; nadir values for hemoglobin, hematocrit, and platelet count; use of dialysis; and length of hospitalization . RESULTS: Twenty-seven of 247 children with HUS had non-enteropathic HUS (11%) . Twenty-four patients (15 boys, 9 girls), whose medical records were complete and available for review, comprise the study cohort . Mean age at onset was 4.2 +/- 0.9 (SE) years . Infection caused by S pneumoniae was diagnosed in 9 patients (38%) . Dialysis was performed in 17 patients (71%) for 40 +/- 27 days . Median length of hospitalization was 22 days (range, 2 to 71 days) . Children with S pneumoniae-related HUS had a longer hospital stay than those with other causes of non-enteropathic HUS, but all patients with S pneumoniae-related HUS recovered kidney function . Dialysis therapy was required more often (17 of 24 versus 59 of 145 children; P = 0.025) and hospital stays were longer (median, 22 versus 9 days; P = 0.002) in children with non-enteropathic HUS compared with patients with D+ HUS who were enrolled in the SYNSORB Pk clinical trial . CONCLUSION: (1) The incidence of non-enteropathic HUS is approximately one tenth that of D+ HUS; (2) patients with non-enteropathic HUS require dialysis therapy more often and are hospitalized more than twice as long during the acute episode compared with those with D+ HUS; (3) infection caused by S pneumoniae accounts for nearly 40% of cases of non-enteropathic HUS; and (4) although S pneumoniae-related HUS is associated with a less favorable short-term course than other types of non-enteropathic HUS or D+ HUS, the long-term prognosis for recovery of renal function appears to be good in these patients.

Curr Opin Infect Dis, 2004 Jun, 17(3), 225 - 9
Bacterial virulence factors in neonatal sepsis: group B streptococcus; Herbert MA et al.; PURPOSE OF REVIEW: Group B streptococcus is a leading cause of neonatal pneumonia, septicaemia and meningitis . Up to one quarter of women in labour are now given intravenous antibiotics to prevent early-onset disease by the organism, a situation that will remain constant until a successful vaccine is available . From a molecular understanding of the pathogenicity of group B streptococcus we may be able to devise novel means for controlling disease, such as identifying inhibitors of key metabolic pathways or regulatory networks . This review summarizes our post-genomic knowledge of the regulation, metabolism and virulence of group B streptococcus . RECENT FINDINGS: Although advances have been made in the understanding of classic group B streptococcus virulence traits, such as capsular polysaccharide, beta-haemolysin, C5a peptidase, adhesins and immunogenic surface proteins, the major recent contribution to group B streptococcus pathogenesis has been the whole genome sequencing of three group B streptococcus strains, representing serotypes Ia, III and V . From these genomes, we not only see where the classic virulence genes map, but we can also gain insights into the metabolism and regulation of the organism and how these affect its virulence . SUMMARY: Knowledge of virulence factors and the organism's metabolism and gene regulation offers opportunities to find novel means of preventing group B streptococcus infection in babies.

Curr Opin Infect Dis, 2004 Jun, 17(3), 217 - 24
Serious bacterial infections in newborn infants in developing countries; Osrin D et al.; PURPOSE OF REVIEW: The overwhelming majority of the world's annual 4 million neonatal deaths occur in developing countries . This review therefore briefly addresses the burden, aetiology, prevention and management of serious neonatal bacterial infections in low-income settings . RECENT FINDINGS: Bacterial infection is the biggest cause of neonatal admissions to hospitals, and probably the biggest cause of morbidity in the community, but its burden is unclear . The commonest serious infections involve bacteraemia, meningitis and respiratory infection, and case fatality rates may be as high as 45% . Key pathogens are Escherichia coli, Klebsiella species, Staphylococcus aureus and Streptococcus pyogenes . The incidence of neonatal infections with group B streptococcus is highly variable, as is the spectrum of antimicrobial resistance . SUMMARY: Current areas of research include the rectification of micronutrient deficiencies, neonatal skin care, appropriate breastfeeding recommendations, cleansing of the birth canal, and simplified methods of diagnosis of infection . Operational activities include the control of neonatal tetanus, the diagnosis and treatment of sexually transmitted infections, integrated strategies for improving pregnancy, childbirth and neonatal survival, community-based management of acute respiratory infections, and community-based management of neonatal sepsis.

Curr Opin Infect Dis, 2004 Jun, 17(3), 177 - 84
Pneumococcal vaccines: do they prevent infection and how?
Posfay-Barbe KM, Wald ER.
PURPOSE OF REVIEW: The purpose of this review is to update and summarize information concerning the epidemiology of infections due to Streptococcus pneumoniae and the early results of effectiveness studies of the heptavalent pneumococcal conjugate vaccine . This vaccine was licensed in the US in 2000, and has been used increasingly since that time . RECENT FINDINGS: Several studies have documented a dramatic decline in the rate of invasive infections due to S . pneumoniae in children under 5 years of age . There has been a simultaneous decrease in serious infections in persons over 20 years of age, presumably because of a decrease in transmission of S . pneumoniae to unvaccinated individuals . Three different studies have shown a modest reduction in the overall number of cases of acute otitis media in children who have received the vaccine . Although the overall number of cases has decreased there has been an increase in the number of cases of acute otitis media caused by serotypes of S . pneumoniae not contained in the vaccine . Worldwide, many studies have appeared that examine the prevalence of different pneumococcal serotypes/serogroups as a cause of invasive and respiratory disease, to assess the likelihood that the vaccine will be effective in particular geographic areas . SUMMARY: Use of the heptavalent pneumococcal conjugate vaccine has led to a major decline in the prevalence of invasive pneumococcal disease (bacteremia, meningitis) and a more modest decrease in respiratory tract infections (acute otitis media, pneumonia) . Continued surveillance is essential to document future trends in the occurrence of pneumococcal infections and the enduring protectiveness of the heptavalent pneumococcal conjugate vaccine.

Am J Med Sci, 2004 May, 327(5), 253 - 4
Deep sternal wound infection caused by group g streptococcus after open-heart surgery; Sarria JC et al.; We report the first case of deep sternal wound infection caused by group G Streptococcus after open-heart surgery . The patient's clinical presentation was nonspecific and his diagnosis was delayed . Surgical debridement and a 4-week course of intravenous antibiotics consisting of sequential penicillin plus gentamicin/ceftriaxone led to recovery . Group G Streptococcus should be suspected as an important postoperative pathogen.

Trends Microbiol, 2004 Jun, 12(6), 288 - 95
Error-prone replication for better or worse; Tippin B et al.; Precise genome duplication requires accurate copying by DNA polymerases and the elimination of occasional mistakes by proofreading exonucleases and mismatch repair enzymes . The commonly held belief that 'if something is worth doing, then it's worth doing well' normally applies to DNA replication and repair, however, there are exceptions . This review describes elements that are crucial to cell fitness, evolution and survival in the recently discovered error-prone DNA polymerases . Large numbers of errant DNA polymerases, spanning microorganisms to humans, are used to rescue stalled replication forks by copying damaged DNA and even undamaged DNA to generate 'purposeful' mutations that generate genetic diversity in times of stress . Here we focus on low-fidelity polymerases from bacteria, comparing Escherichia coli, archeabacteria and those most recently discovered in Gram-positive Bacilli, Streptococcus, pathogenic Mycobacterium and intein-containing cyanobacteria.

J Clin Immunol, 2004 Jul, 24(4), 411 - 7
Antibody response to a seven-valent pneumococcal conjugated vaccine in patients with ataxia-telangiectasia; Sanal O et al.; Immunodeficiency is a characteristic feature of ataxia-telangiectasia (A-T) . Humoral immunodeficiency generally consists of hypogammaglobulinemia and impaired antibody response to bacterial and viral antigens . We previously observed defective antibody response to 23-valent pneumococcal polysaccharide vaccine (PPV) in 96% of 29 patients with A-T . In this study, we investigated the antibody response to a seven-valent pneumococcal conjugate vaccine, PCV7, in 14 patients with A-T . IgG antibody levels to four pneumococcal serotypes, 6B, 14, 19F, 23F, which were included in PCV7, were measured by ELISA in pre- and postimmunization serum samples . Antibody titers against each individual Streptococcus pneumoniae serotype was considered to be positive when serotype specific pneumococcal antibody titer was higher than 10% (>10 U/mL) of the reference plasma pool level . However, when the fold increase (FI) in postimmunization antibody titer was less than two, the subject was determined to be unresponsive to the given serotype . The values were compared with the results obtained in age- and ethnic-matched children after one dose of PPV . Only two patients produced antibodies to one serotype each; one to serotype 19 with a fold increase of <2, and the other to serotype 23F with a fold increase of 5.7 based on the above criteria, although the differences between pre- and postvaccine antibody titers for serotypes 14, 19, and 23 appeared to be statistically significant . In conclusion, A-T patients failed to respond to one dose of PCV7 vaccine . Two or more doses of conjugated vaccine may be required to recruit the help of T lymphocytes in A-T patients.

Indian J Pediatr, 2004 May, 71(5), 423 - 6
Streptococcus pyogenes meningitis; Mathur P et al.; Group A Streptococcus (GAS) is a rare cause of meningitis . Although it has a high mortality, the condition is easily treatable if diagnosed early since the bacteria retains its sensitivity to many antimicrobials . The authors report here two cases of GAS meningitis along with a review of world literature.

Rev Biol Trop, 2003 Sep-Dec, 51(3-4), 629 - 33
Polythene and plastic-degrading microbes in an Indian mangrove soil; Kathiresan K; Biodegradation of polythene bags and plastic cups was analyzed after 2, 4, 6, and 9 months of incubation in the mangrove soil . The biodegradation of polythene bags was significantly higher (up to 4.21% in 9 months) than that of plastic cups (up to 0.25% in 9 months) . Microbial counts in the degrading materials were recorded up to 79.67 x 10(4) per gram for total heterotrophic bacteria, and up to 55.33 x 10(2) per gram for fungi . The microbial species found associated with the degrading materials were identified as five Gram positive and two Gram negative bacteria, and eight fungal species of Aspergillus . The species that were predominant were Streptococcus, Staphylococcus, Micrococcus (Gram +ve), Moraxella, and Pseudomonas (Gram -ve) and two species of fungi (Aspergillus glaucus and A . niger) . Efficacy of the microbial species in degradation of plastics and polythene was analyzed in shaker cultures . Among the bacteria, Pseudomonas species degraded 20.54% of polythene and 8.16% of plastics in one-month period . Among the fungal species, Aspergillus glaucus degraded 28.80% of polythene and 7.26% of plastics in one-month period . This work reveals that the mangrove soil is a good source of microbes capable of degrading polythene and plastics.

J Infect Chemother, 2004 Apr, 10(2), 115 - 20
Macrolide-resistant genes of Streptococcus pyogenes isolated from the upper respiratory tract by polymerase chain reaction; Billal DS et al.; The growing number of macrolide-resistant strains of Streptococcus pyogenes is an increasing problem worldwide . This study evaluated 300 clinical isolates obtained from the upper respiratory tract . Minimal inhibitory concentrations (MICs) of erythromycin (EM), azithromycin (AZM), and clindamycin (CLDM), serotypes, and macrolide resistance genes of mefA, ermB, and ermTR were determined . The genetic relationship of EM-resistant and susceptible strains were also analyzed by pulsed-field gel electrophoresis (PFGE) . Twenty-nine (9.7%) EM-resistant S . pyogenes were identified . Of the 29 strains showing resistance to EM, 22 isolates (7.3%, MIC 3.13-12.5 microg/ml) expressed the mefA gene . The predominant serotypes among the mefA-positive isolates were T12, emm9 or T25, emm75-1 . The two isolates (0.1%) that possessed the ermB gene were highly resistant to EM (MIC > 100 microg/ml) . The remaining five strains (1.6%) possessed the ermTR gene (MIC 3.13-100 microg/ml) . Restriction fragment polymorphism analyzed by pulsed-field gel electrophoresis (PFGE) by SmaI and ApaI digestions showed several clones among the mefA-positive S . pyogenes . Our findings suggest that the mefA gene is the predominant mechanism for macrolide resistance and that this gene is horizontally transmitted among M phenotype strains of S . pyogenes . Consequently, macrolides would not be the first drug of choice for treatment of tonsillitis and other S . pyogenes-related diseases . Physicians and researchers need to take into consideration the macrolide resistance of some strains of S . pyogenes.

J Infect Chemother, 2004 Apr, 10(2), 105 - 9
The role of the capsule of the Streptococcus milleri group in its pathogenicity; Kanamori S et al.; Study of the pathogenicity of encapsulated strains of the Streptococcus milleri group (SMG) was performed by examination of the ability to cause subcutaneous abscesses in mice and by phagocytosis and phagocytic killing of human polymorphonuclear neutrophils (PMNs) against the organisms . All 3 encapsulated isolates from patients with pneumonia or lung abscess induced abscesses in the mice; however, only 2 of 20 unencapsulated isolates from patients with lung abscess or thoracic empyema did so . The 3 encapsulated strains inhibited more phagocytosis and phagocytic killing of PMNs than the unencapsulated strains . In addition, encapsular material separated from Streptococcus constellatus RZYK001 also inhibited phagocytosis and phagocytic killing in proportion to increasing concentrations of the capsular material . These results suggest that capsular material produced by SMG might be a pathogenic factor.

Shanghai Kou Qiang Yi Xue, 1996 Jun, 5(2), 89 - 90
{A study on the influence of zinc on the growth and metabolism of streptococcus in an artificial mouth model}; Zhu M et al.; The influence of zinc on the growth and metabolism of Streptococcus was studied under controlled conditions in an artificial mouth model.the results confirmed the previous conclusion that zinc was capable of inhibiting the growth and metabolism of streptococcus both vivo and in vitro.At the same time,the feasibility of the imitative experiments in an artificial mouth model was proved.

Shanghai Kou Qiang Yi Xue, 1996 Dec, 5(4), 219 - 21
{Environmental factors in oral cavity effect on growing of Streptococcus sanguis and Strepococcus mutans-Oxygen of effects}; Xiang XL et al.; Oral Streptococcus species belong to facultative aerobes,and are easily inhibited by metabolic products of oxygen.Our study uses continues culture technique through adding oxygen in order to observe growing states of S.sanguis 34 and S.mutans Ingbritt(c) The result is that the resistance to oxygen of S.sanguis 34 is stronger than S.mutans Ingbritt(c).This result is in accordance with the phenomenon that S.sanguis is a pioneer colonizing germ of dental plaque.The lower resistance to oxygen of S.mutans indicates that the growth and cariogenic action of S.mutans are dependent on the internal circumstance of plaque.

Shanghai Kou Qiang Yi Xue, 1996 Dec, 5(4), 215 - 8
{Environmental factors in oral cavity effect on growing of Streptococcus sanguis and Streptococcus mutans-effect of Mucin}; Xiang XL et al.; Our study use continues culture technique through adding mucin as the limiting element in chemically defined culture,medium in order to observe growing states of S.sanguis 34 and,S.mutants Ingbritt(c).The results as follows:S.sanguis 34 were able to grow as pure culture on mucin;S.mutans was unable to grow as a pure culture on mucin,but attained a significant population size in the presence of S.sanguis.These results indicate a synergetic degradation of mucin by S.sanguis and S.mutans in mixed chemism culture.

Shanghai Kou Qiang Yi Xue, 1996 Dec, 5(4), 210 - 4
{A comparative analysis of biological properties of glucosylrtanterases secreted by Streptococcus sanguis and Streptococcus mutans}; Xiang XL et al.; Our study make a comparative analysis of biological properties of glucosyltranterases(GTF) secreted by S.sanguis 34 and S.mutans Ingbritt(c).The results as follows:(1)S.sanguis 34 has more extractions of extracelluar GTF than S.mutans Ingbritt(c) under equal cultured conditions,but S.mutans Inggbritt(c) has more activity of extracelluar GTF than S.sanguis.These differences are identical with the difference of their cariogenicities;(2)GTF of S.sanguis 34 can also synthesize water-soluble glucan,and is a multienzyme complex system as S.mutans . S.sanguis might catalyse sufficient dextran formation to stimulate the primer-dependent activity of the S.mutans GTF enzyme . This might aid the adhesion of this comparatively late colonizer;(3)easier to obstruct high pure GTF of S.sangusi 34 S.mutans.

Shanghai Kou Qiang Yi Xue, 1993 Dec, 2(4), 214 - 7
{Purification and characterization of lipoteichoic acid from Streptococcus Sobronus}; Li MY et al.; Lipoteichoic acid (LTA) is amphipathic molecule found in cell surface of gram-positive bacteria.Although there were some hypothese about adhesion of LTA,the role of LTA in bacterial pathogenicety is still unclear.For this reason,a seria studies of LTA were carried out.In this study LTA was extracted by hot aqueous phenol from Strptococcus Sobronus 6715 grown in tryptic soy both supplemented with 5% sucrose for 17 hours,and was purfied on gel filtration by Sepharose 6B,chloroform-methanol precipitation and chromtography on DEAE-Sephacel.The purified LTA was characterized by FT-IR and 1H-NMR with standard sample of LTA (Sigma Co) as control.The results showed that LTA contains alanine ester and glycosyl residues which take important role in the amphipathic molecule.

Acta Trop, 2004 Jun, 91(1), 53 - 68
Zebrafish as a model host for streptococcal pathogenesis; Miller JD et al.; Streptococcal pathogens continue to evade concerted efforts to determine clear-cut virulence mechanisms, although numerous genes have been implicated in pathogenesis . A single species can infect a diversity of tissues, suggesting the expression of specific virulence factors based on the local tissue environment or stage of infection . In an effort to identify the interactions that occur between the host and pathogen that lead to activation of virulence mechanisms and contribute to specific streptococcal disease states, we have developed a unique animal model, the zebrafish (Danio rerio), to characterize specific virulence mechanisms utilized within various tissues in vivo . We are using this model host to study infection by two streptococcal species that represent two forms of streptococcal disease: a natural pathogen of fish and humans, Streptococcus iniae and a human-specific pathogen, Streptococcus pyogenes . S . iniae primarily causes a fatal systemic disease in the zebrafish following intra-muscular injection, with similar pathologies to that seen in human infections caused by Streptococcus agalactiae and S . pneumoniae . While the fatal infection by S . pyogenes causes a locally spreading necrotic disease confined to the muscle with pathology similar to what is observed in a human infection of necrotizing fasciitis . By studying pathogens that are virulent for both fish and humans and that mediate disease states in the zebrafish that are identical to those found in human streptococcal infections, we will be able to identify common virulence strategies shared by a number of Gram-positive pathogens.

Clin Infect Dis, 2004 May 15, 38(10), 1394 - 400 Epub 2004 Apr 28.
Streptococcus bovis endocarditis and its association with chronic liver disease: an underestimated risk factor; Tripodi MF et al.; Clinical and epidemiological characteristics of Streptococcus bovis endocarditis were prospectively studied among 199 patients with definite endocarditis . Thirty patients (15.1%) had S . bovis endocarditis . Compared with patients with non-S . bovis endocarditis, these 30 patients were older (mean age, 58.6+/-12.4 years vs . 46.0+/-17.0 years; P<.001) and had higher rates of bivalvular involvement (43.3% vs . 7.7%; P<.001), embolism (73.3% vs . 40.2%; P=.002), and diskitis (23.3% vs . 0.6% P<.001) . In patients with S . bovis biotype I (S . bovis I) endocarditis, advanced liver disease was present in 56.7%, compared with 15.3% of patients with non-S . bovis endocarditis (P<.001), and colonic adenoma was present in 46.7% . The in-hospital mortality rate (16.7%) was correlated with delayed diagnosis and advanced liver diseases . In our city, S . bovis I endocarditis is frequently correlated with liver diseases; diskitis may be the first sign of the disease.

Pediatr Res, 2004 Jun, 55(6), 966 - 71
Defective neutrophil oxidative burst in preterm newborns on exposure to coagulase-negative staphylococci; Bjorkqvist M et al.; The neutrophil oxidative burst is a product of the regulated assembly of the multicomponent oxidase enzyme . Our aim was to compare the oxidative burst in term (n = 10) and preterm newborns <31 wk gestational age (n = 10) after stimulation with coagulase-negative staphylococci in vitro . Strains of Streptococcus epidermidis with different invasive and slime-producing properties, one strain of S . haemolyticus, and one strain of group B-streptococcus were investigated . A whole-blood flow cytometric assay using the oxidation of hydroethidine to ethidium bromide was used . The oxidative activity in unstimulated neutrophil granulocytes {polymorphonuclear leukocytes (PMNLs)} was similar in term and preterm newborns, but the preterm newborns showed a significantly lower capacity to up-regulate the oxidative burst intensity after bacterial stimulation (p = 0.004) . In the term but not in the preterm group, the oxidative burst intensity after bacterial stimulation correlated with the baseline oxidative burst intensity . After bacterial stimulation, there was a trend toward a greater percentage of activated neutrophils in the term group than in the preterm group, but the difference was less pronounced than that in oxidative burst intensity . Significant differences in oxidative burst response to different bacterial strains were observed (p < 0.001), but the differences could not be correlated exclusively to invasive capacity or slime-producing properties . It is concluded that the baseline oxidative activity is similar in term and preterm PMNLs but that preterm PMNLs have a decreased capacity to increase the oxidative burst in response to bacterial stimulation.

Infect Immun, 2004 Jun, 72(6), 3668 - 73
Role of Streptococcus pyogenes two-component response regulators in the temporal control of Mga and the Mga-regulated virulence gene emm; Ribardo DA et al.; We examined the role of Streptococcus pyogenes two-component response regulators (SptR) in expression of Mga and the Mga-regulated gene emm . Both serotype M6 and serotype M1 mutants in 12 of the 13 identified sptR genes exhibited levels of emm transcripts and Mga protein comparable to those of the wild type during exponential and stationary phases of growth . Thus, temporal control of these virulence genes does not require Spt response regulators.

Infect Immun, 2004 Jun, 72(6), 3584 - 91
Role of HtrA in the virulence and competence of Streptococcus pneumoniae; Ibrahim YM et al.; HtrA is a major virulence factor of Streptococcus pneumoniae (the pneumococcus) . Deletion of the gene for HtrA from strain D39 of the pneumococcus completely abolished its virulence in mouse models of pneumonia and bacteremia, while the virulence of a second strain (TIGR4) was dramatically reduced . HtrA-negative mutants induced much less inflammation in the lungs during pneumonia than the wild type . HtrA is involved in the ability of the pneumococcus to grow at high temperatures, to resist oxidative stress, and to undergo genetic transformation . The expression and cellular location of several known virulence factors of the pneumococcus were not affected by the lack of HtrA.

Infect Immun, 2004 Jun, 72(6), 3505 - 14
Somatic hypermutation and diverse immunoglobulin gene usage in the human antibody response to the capsular polysaccharide of Streptococcus pneumoniae Type 6B; Zhou J et al.; Combinatorial cloning and expression library analysis were used to determine the expressed human antibody repertoire specific for the capsular polysaccharide (PS) of Streptococcus pneumoniae serotype 6B . Sequence analysis of 55 6B-specific antibody Fab fragments isolated from six vaccinated donors reveal that different individuals used a variety of heavy and light chain germ line variable (V) region genes to form pneumococcal capsular PS (PPS) 6B-specific paratopes . Within each donor, however, the response was more restricted, with five of the six donors using at most one or two gene pairs to form combining sites . Analysis also indicated that although the response in each donor was oligoclonal in terms of variable gene usage, the combination of extensive somatic hypermutation, deletion of germ line-encoded residues, insertion of non-germ line-encoded residues, and intraclonal isotype switching generated a surprising degree of paratope diversity within the individuals analyzed . In contrast to previously studied PS-specific responses, we find that the PPS 6B repertoire makes use of a diverse collection of heavy-chain and light-chain V region gene products to form specific paratopes, with no apparent tendency for conservation of immunoglobulin gene usage between individuals.

Infect Immun, 2004 Jun, 72(6), 3495 - 504
The novel fibrinogen-binding protein FbsB promotes Streptococcus agalactiae invasion into epithelial cells; Gutekunst H et al.; Streptococcus agalactiae is a major cause of bacterial sepsis and meningitis in human newborns . The interaction of S . agalactiae with host proteins and the entry into host cells thereby represent important virulence traits of these bacteria . The present report describes the identification of the fbsB gene, encoding a novel fibrinogen-binding protein that plays a crucial role in the invasion of S . agalactiae into human cells . In Western blots and enzyme-linked immunosorbent assay (ELISA) experiments, the FbsB protein was demonstrated to interact with soluble and immobilized fibrinogen . Binding studies showed the N-terminal 388 residues of FbsB and the Aalpha-subunit of human fibrinogen to recognize each other . By reverse transcription (RT)-PCR, the fbsB gene was shown to be cotranscribed with the gbs0851 gene in S . agalactiae . Deletion of the fbsB gene in the genome of S . agalactiae did not influence the binding of the bacteria to fibrinogen, suggesting that FbsB does not participate in the attachment of S . agalactiae to fibrinogen . In tissue culture experiments, however, the fbsB deletion mutant was severely impaired in its invasion into lung epithelial cells . Bacterial invasion could be reestablished by introducing the fbsB gene on a shuttle plasmid into the fbsB deletion mutant . Furthermore, treatment of lung epithelial cells with FbsB fusion protein blocked S . agalactiae invasion of epithelial cells in a dose-dependent fashion . These results suggest an important role of the FbsB protein in the overall process of host cell entry by S . agalactiae.

Infect Immun, 2004 Jun, 72(6), 3331 - 5
Age-specific immunoglobulin g (IgG) and IgA to pneumococcal protein antigens in a population in coastal kenya; Laine C et al.; Streptococcus pneumoniae is the primary etiological agent of community-acquired pneumonia and a major cause of meningitis and bacteremia . Three conserved pneumococcal proteins-pneumolysin, pneumococcal surface adhesin A (PsaA), and pneumococcal surface protein A (PspA)-are currently being investigated as vaccine candidates . Such protein-based vaccines, if proven effective, could provide a cheaper alternative to conjugate vaccine formulae . Few data from sub-Saharan Africa exist concerning the development of natural antibody to these antigens, however . To investigate the age-specific development of antiprotein immunoglobulin G (IgG) and IgA antibody responses, the sera of 220 persons 2 weeks to 84 years of age from coastal Kenya were assayed using enzyme-linked immunosorbent assays . IgG and IgA antibody responses to each antigen were observed in all age groups . Serum concentrations of IgG and IgA antibody responses to PspA and PdB (a recombinant toxoid derivative of pneumolysin), but not to PsaA, increased significantly with age (P < 0.001) . No decline was observed in the sera of the elderly . Anti-protein IgG concentrations were only weakly correlated (0.30 < r < 0.56; P < 0.0001), as were IgA concentrations (0.24 < r < 0.54; P < 0.0001).

Infect Immun, 2004 Jun, 72(6), 3228 - 36
Recombinant Streptococcus equi proteins protect mice in challenge experiments and induce immune response in horses; Flock M et al.; Horses that have undergone infection caused by Streptococcus equi subspecies equi (strangles) were found to have significantly increased serum antibody titers against three previously characterized proteins, FNZ (cell surface-bound fibronectin binding protein), SFS (secreted fibronectin binding protein), and EAG (alpha2-macroglobulin, albumin, and immunoglobulin G {IgG} binding protein) from S . equi . To assess the protective efficacy of vaccination with these three proteins, a mouse model of equine strangles was utilized . Parts of the three recombinant proteins were used to immunize mice, either subcutaneously or intranasally, prior to nasal challenge with S . equi subsp . equi . The adjuvant used was EtxB, a recombinant form of the B subunit of Escherichia coli heat-labile enterotoxin . It was shown that nasal colonization of S . equi subsp . equi and weight loss due to infection were significantly reduced after vaccination compared with a mock-vaccinated control group . This effect was more pronounced after intranasal vaccination than after subcutaneous vaccination; nearly complete eradication of nasal colonization was obtained after intranasal vaccination (P < 0.001) . When the same antigens were administered both intranasally and subcutaneously to healthy horses, significant mucosal IgA and serum IgG antibody responses against FNZ and EAG were obtained . The antibody response was enhanced when EtxB was used as an adjuvant . No adverse effects of the antigens or EtxB were observed . Thus, FNZ and EAG in conjunction with EtxB are promising candidates for an efficacious and safe vaccine against strangles.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 2244 - 50
Complete sequences of six penicillin-binding protein genes from 40 Streptococcus pneumoniae clinical isolates collected in Japan; Sanbongi Y et al.; All six penicillin-binding protein (PBP) genes, namely, pbp1a, pbp1b, pbp2a, pbp2b, pbp2x, and pbp3, of 40 Streptococcus pneumoniae clinical isolates, including penicillin-resistant S . pneumoniae isolates collected in Japan, were completely sequenced . The MICs of penicillin for these strains varied between 0.015 and 8 microg/ml . In PBP 2X, the Thr550Ala mutation close to the KSG motif was observed in only 1 of 40 strains, whereas the Met339Phe mutation in the STMK motif was observed in six strains . These six strains were highly resistant (MICs >/= 2 microg/ml) to cefotaxime . The MICs of cefotaxime for 27 strains bearing the Thr338Ala mutation tended to increase, but the His394Leu mutation next to the SSN motif did not exist in these strains . In PBP 2B, the Thr451Ala/Phe/Ser and Glu481Gly mutations close to the SSN motif were observed in 24 strains, which showed penicillin resistance and intermediate resistance, and the Thr624Gly mutation close to the KTG motif was observed in 2 strains for which the imipenem MIC (0.5 microg/ml) was the highest imipenem MIC detected . In PBP 1A, the Thr371Ser/Ala mutation in the STMK motif was observed in all 13 strains for which the penicillin MICs were >/=1 microg/ml . In PBP 2A, the Thr411Ala mutation in the STIK motif was observed in one strain for which with the cefotaxime MIC (8 microg/ml) was the highest cefotaxime MIC detected . On the other hand, in PBPs 1B and 3, no mutations associated with resistance were observed . The results obtained here support the concept that alterations in PBPs 2B, 2X, and 1A are mainly involved in S . pneumoniae resistance to beta-lactam antibiotics . Our findings also suggest that the Thr411Ala mutation in PBP 2A may be associated with beta-lactam resistance.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 2206 - 13
Short- and long-term effects of pneumococcal conjugate vaccination of children on penicillin resistance; Temime L et al.; Recent observations have shown that wide-scale vaccination with pneumococcal conjugate vaccines was associated with a reduction in invasive disease, supporting the expectation that vaccination could help reduce carriage of Streptococcus pneumoniae and control the spread of resistant strains . However, it is too early to assess whether these effects can be sustained in the long term . Here, we used mathematical modeling to investigate time changes in pneumococcal colonization and resistance induced by conjugate vaccination in an environment where antibiotic exposure is high and resistance is widespread . According to model predictions, vaccination induced a decrease in carriage of vaccine-type pneumococci to very low levels, typically in 10 to 15 years under epidemiologically realistic conditions . Almost simultaneously, non-vaccine-type pneumococci spread in the community . Consequently, while there was a short-term decrease in the overall carriage rate, it was followed after a few years by a renewed, although limited, increase . Vaccination with a heptavalent vaccine did not affect the extent to which antibiotic resistance was selected: in all cases, the distribution of resistance levels peaked at high levels (MIC > 2 microg/ml) after 20 years . With a vaccine optimally designed to include all serotypes currently exhibiting decreased susceptibility to penicillin G, the selection of resistance was slowed down, although not prevented . These results suggest that because of serotype replacement, the effects of vaccination observed today may not be sustained in the long term . As a consequence, vaccination alone may not be successful in controlling selection for resistance in S . pneumoniae.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 2108 - 15
Ciprofloxacin dimers target gyrase in Streptococcus pneumoniae; Gould KA et al.; We have examined the antipneumococcal activities of novel quinolone dimers in which ciprofloxacin was tethered to itself or to pipemidic acid by linkage of C-7 piperazinyl rings . Symmetric 2,6-lutidinyl- and trans-butenyl-linked ciprofloxacin dimers (dimers 1 and 2, respectively) and a pipemidic acid-ciprofloxacin dimer (dimer 3) had activities against Streptococcus pneumoniae strain 7785 that were comparable to that of ciprofloxacin, i.e., MICs of 2, 1, and 4 to 8 microg/ml versus an MIC of 1 to 2 microg/ml, respectively . Surprisingly, unlike ciprofloxacin (which targets topoisomerase IV), several lines of evidence revealed that the dimers act through gyrase in S . pneumoniae . First, ciprofloxacin-resistant parC mutants of strain 7785 remained susceptible to dimers 1 to 3, whereas a gyrA mutation conferred a four- to eightfold increase in the dimer MIC but had little effect on ciprofloxacin activity . Second, dimer 1 selected first-step gyrA (S81Y or S81F) mutants (MICs, 8 to 16 microg/ml) that carried wild-type topoisomerase IV parE-parC genes . Third, dimers 1 and 2 promoted comparable DNA cleavage by S . pneumoniae gyrase and topoisomerase IV, whereas ciprofloxacin-mediated cleavage was 10-fold more efficient with topoisomerase IV than with gyrase . Fourth, the GyrA S81F and ParC S79F enzymes were resistant to dimers, confirming that the resistance phenotype is largely silent in parC mutants . Although a dimer molecule could bind very tightly by bridging quinolone binding sites in the enzyme-DNA complex, the greater potency of ciprofloxacin against gyrase and topoisomerase IV suggests that dimers 1 to 3 bind in a monomeric fashion . The bulky C-7 side chain may explain dimer targeting of gyrase and activity against efflux mutants . Tethered quinolones have potential as mechanistic tools and as novel antimicrobial agents.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 2101 - 7
High prevalence of antimicrobial resistance among clinical Streptococcus pneumoniae isolates in Asia (an ANSORP study); Song JH et al.; A total of 685 clinical Streptococcus pneumoniae isolates from patients with pneumococcal diseases were collected from 14 centers in 11 Asian countries from January 2000 to June 2001 . The in vitro susceptibilities of the isolates to 14 antimicrobial agents were determined by the broth microdilution test . Among the isolates tested, 483 (52.4%) were not susceptible to penicillin, 23% were intermediate, and 29.4% were penicillin resistant (MICs >/= 2 mg/liter) . Isolates from Vietnam showed the highest prevalence of penicillin resistance (71.4%), followed by those from Korea (54.8%), Hong Kong (43.2%), and Taiwan (38.6%) . The penicillin MICs at which 90% of isolates are inhibited (MIC(90)s) were 4 mg/liter among isolates from Vietnam, Hong Kong, Korea, and Taiwan . The prevalence of erythromycin resistance was also very high in Vietnam (92.1%), Taiwan (86%), Korea (80.6%), Hong Kong (76.8%), and China (73.9%) . The MIC(90)s of erythromycin were >32 mg/liter among isolates from Korea, Vietnam, China, Taiwan, Singapore, Malaysia, and Hong Kong . Isolates from Hong Kong showed the highest rate of ciprofloxacin resistance (11.8%), followed by isolates from Sri Lanka (9.5%), the Philippines (9.1%), and Korea (6.5%) . Multilocus sequence typing showed that the spread of the Taiwan(19F) clone and the Spain(23F) clone could be one of the major reasons for the rapid increases in antimicrobial resistance among S . pneumoniae isolates in Asia . Data from the multinational surveillance study clearly documented distinctive increases in the prevalence rates and the levels of antimicrobial resistance among S . pneumoniae isolates in many Asian countries, which are among the highest in the world published to date.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 2037 - 42
Tn2009, a Tn916-like element containing mef(E) in Streptococcus pneumoniae; Del Grosso M et al.; The association between the macrolide efflux gene mef(E) and the tet(M) gene was studied in two clinical strains of Streptococcus pneumoniae that belonged to serotypes 19F and 6A, respectively, and that were resistant to both tetracycline and erythromycin . The mef(E)-carrying element mega (macrolide efflux genetic assembly; 5,511 bp) was found to be inserted into a Tn916-like genetic element present in the chromosomes of the two pneumococcal strains . In both strains, mega was integrated at the same site, an open reading frame identical to orf6 of Tn916 . The new composite element, Tn2009, was about 23.5 kb and, with the exception of the tet(M)-coding sequence, appeared to be identical in both strains . By sequencing of the junction fragments of Tn2009 at the site of insertion into the chromosome, it was possible to show that (i) the insertion site was identical in the two clinical strains and (ii) the integration of Tn2009 caused a 9.5 kb-deletion in the pneumococcal chromosome . It was not possible to detect the conjugal transfer of Tn2009 to a recipient pneumococcal strain; however, transfer of the whole element by transformation was shown to occur . It is possible to hypothesize that Tn2009 relies on transformation for its spread among clinical strains of S . pneumoniae.

J Mol Microbiol Biotechnol, 2003, 6(3-4), 155 - 63
Molecular characterization of the essential response regulator protein YycF in Bacillus subtilis; Watanabe T et al.; The response regulator YycF is essential for cell growth in gram-positive bacteria including Bacillus subtilis, Staphylococcus aureus and Streptococcus pneumoniae . To study the function of YycF in the essential process, we characterized a YycF (H215P) mutation that caused temperature-sensitive growth in B . subtilis . The response regulators YycF and YycF (H215P) were analyzed using circular dichroism spectroscopy, whose T(m) values were 56.0 and 45.9 degrees C, respectively, suggesting that YycF (H215P) significantly affects the protein structure with an increase in temperature . Furthermore, using the gel mobility shift assay and DNase I footprinting, we investigated the effect of YycF (H215P) on binding to the YycF box of ftsAZ operon of B . subtilis . The replacement of the histidine 215 with proline resulted in a decrease of the DNA-binding ability of YycF in vitro . In vivo, using Escherichia coli two-hybrid and homodimerization assays, we clarified that His 215 of YycF plays a crucial role in the homodimerization of the protein . Thus the essential genes involved in growth of B . subtilis appear to be regulated by the homodimer of YycF . These results suggest that the YycF dimerization is an excellent target for the discovery of novel antibiotics .

Kurume Med J, 2004, 51(1), 53 - 7
Clinical features of acute respiratory infections associated with the Streptococcus milleri group in the elderly; Sugihara E et al.; The Streptococcus milleri group are becoming increasingly recognized as important pulmonary pathogens which may lead to the development of empyema or lung abscesses . Although several small series have been reported, the clinical and laboratory features of Streptococcus milleri infection have yet to be fully characterized in the elderly . We retrospectively examined the clinical features of 19 patients with Streptococcus milleri pulmonary disease who were admitted to our hospital between 2000 and 2002, based on their clinical records and laboratory data . The microbiological diagnosis was based on the results of quantitative sputum culture and other invasive procedures, including transthoracic needle aspiration or bronchoscopic examinations . There were thirteen cases of pneumonia, two of contaminant pneumonia and pleuritis, one of bronchitis, two of pulmonary abscess, and one of empyema . The patients ranged in age from 65 to 91 . The most common symptoms at presentation were shortness of breath, coughing, sputum, and weight loss . An underlying disease existed in 14 of the 19 cases . We conclude that the Streptococcus milleri group is a more important cause of pulmonary infections than has been previously recognized.

J Med Microbiol, 2004 Jun, 53(Pt 6), 505 - 8
Equivalence of high-virulence clonotypes of serotype III group B Streptococcus agalactiae (GBS); Fleming KE et al.; Analysis of growth characteristics, multilocus enzyme electrophoresis, restriction digest pattern (RDP) typing and multilocus sequence typing have identified clonotypes of serotype III group B Streptococcus agalactiae (GBS) associated with invasive infection in neonates . This study sought to unify phenotypic and genotypic classifications of type III GBS strains associated with increased virulence in newborns . High-virulence clonotype (HVC) strains possessed the translation initiation factor 2 (infB) C allele, found in RDP type III-3 strains, and hybridized with the RDP type III-3-specific probe AA3.6, whereas non-HVC strains shared the infB A allele and genomic DNA from these strains did not hybridize with the AA3.6 probe . The characteristic growth lag of HVC GBS at 40 degrees C has been attributed to the presence of a heat-labile fructose-1,6-bisphosphate aldolase (Fba) enzyme in these strains . The deduced amino acid sequence of fba genes of both HVC and non-HVC strains, however, were identical . HVC and RDP type III-3 represent the same genetically related group of bacteria . The characteristic growth differences of virulent strains of type III GBS, however, are not directly attributable to differences in fba.

J Bacteriol, 2004 Jun, 186(11), 3447 - 52
Single-step capsular transformation and acquisition of penicillin resistance in Streptococcus pneumoniae; Trzcinski K et al.; The capsule (cps) locus of Streptococcus pneumoniae is flanked by the pbp2x and pbp1a genes, coding for penicillin-binding proteins, enzymes involved in cell wall synthesis that are targets for beta-lactams . This linkage suggested to us that selection for beta-lactam resistance might coselect for capsular transformants . The recombination event would then involve PBP genes, as well as the cps operon, and would change both the serotype and the resistance profile of the strain . We transformed beta-lactam-susceptible strain TIGR4 by using whole genomic DNA extracted from multidrug-resistant strain GA71, a serotype 19F variant of pneumococcal clone Spain(23F)-1, and selected beta-lactam-resistant transformants . Smooth colonies appearing on selective plates were subcultured, serotyped by the Quellung reaction, and genotyped to confirm the presence of the GA71 pbp2x-cps19-pbp1a locus in the TIGR4 genetic background by restriction fragment length polymorphism analysis of the whole locus and its flanking regions . The results showed that a new serotype, combined with resistance to beta-lactams, could emerge in a susceptible strain via a single transformation event . Quantitative analysis showed that transfer of the cps locus had occurred at an elevated rate in beta-lactam-selected transformants . This suggests that in natural settings selection by host immunity and selection by antibiotics may be interrelated because of "hitchhiking" effects due to linkage of resistance determinants and the capsule locus.

J Antimicrob Chemother, 2004 Jun, 53 Suppl 2, ii59 - 66
Ertapenem versus ceftriaxone for the treatment of community-acquired pneumonia in adults: combined analysis of two multicentre randomized, double-blind studies; Ortiz-Ruiz G et al.; The efficacy and safety of ertapenem, 1 g once a day, for the treatment of community-acquired pneumonia (CAP) requiring parenteral therapy were compared with those of ceftriaxone, 1 g once a day, in 866 hospitalized adults randomized in two prospective, double-blind, multicentre studies . Patients were stratified according to Pneumonia Severity Index (< or = 3 or >3) or age (< or = 65 or >65 years) . After > or = 3 days of parenteral antimicrobial therapy, patients who had clinically improved could be switched to oral co-amoxiclav . The median durations of parenteral, oral and total therapy in the 658 clinically evaluable patients, of whom 88% were switched to oral therapy, were 4, 7 and 12 days, respectively, in both treatment groups . The most common pathogen was Streptococcus pneumoniae, of which 79% (143/181) were penicillin susceptible and 3.3% (6/181; three in each treatment group) were penicillin resistant . Cure rates for the two treatments were equivalent: 91.9% for ertapenem and 92.0% for ceftriaxone (95% confidence interval for the difference, adjusted for strata: -4.5 to 4.4) . Cure rates in the different severity and age strata and bacterial eradication rates for both treatment groups were also similar . The most common drug-related adverse events in both treatment groups were diarrhoea and mild-to-moderate elevations in aminotransferase levels . The results of these studies demonstrate that ertapenem, 1 g once a day, was highly effective therapy for CAP in hospitalized adults with moderate-to-severe disease.

J Antimicrob Chemother, 2004 Jul, 54(1), 122 - 9 Epub 2004 May 18.
Molecular characterization of penicillin non-susceptible Streptococcus pneumoniae in Christchurch, New Zealand; Bean DC et al.; OBJECTIVES: To determine the epidemiological relationship between non-invasive penicillin non-susceptible Streptococcus pneumoniae isolates collected in the Christchurch community between 1997 and 2001 . METHODS: One hundred and ninety-seven pneumococcal isolates were examined by macrorestriction profile analysis of SmaI-digested genomic DNA separated by PFGE and restriction fragment length polymorphism analysis of penicillin binding protein genes . RESULTS: Four major clonal lineages were identified, the largest and most homogeneous containing 95 (48.2%) of the isolates, the bulk of which (93.7%), had identical macrorestriction patterns . Members of this clonal group were multidrug-resistant and exhibited high resistance to third-generation cephalosporins, with MICs > or =8.0 mg/L not uncommon (23.1%) . Two of the clonal groups, each containing 24 (12.2%) isolates, appeared indistinguishable from the globally widespread Spain23F-1 and France9V-3 strains, respectively . The fourth (12.7% of isolates) multidrug-resistant clone possessed intermediate penicillin susceptibility (MIC 0.12 mg/L) . CONCLUSIONS: This study shows that several distinct penicillin-resistant pneumococcal clones are present in the Christchurch community, most of which appear to have been imported into New Zealand.

Vaccine, 2004 Jun 2, 22(17-18), 2209 - 20
Pneumococcal vaccines: an update on current strategies; Bogaert D et al.; Streptococcus pneumoniae is a major cause of morbidity and mortality in infants, children and the elderly . Despite the availability of excellent antimicrobial therapy and adequate health care systems, respiratory diseases and invasive infections caused by pneumococci still comprise a major health problem . The emerging resistance to penicillin and other commonly used antibiotics underscores the importance of the development of novel vaccine strategies to combat pneumococcal disease . Although the 23-valent polysaccharide (PS) vaccine is immunogenic and protective in most adults and children over 5 years of age, they fail to protect children under 2 years of age . Fortunately, the recent conjugate vaccines have shown to be highly efficacious in preventing invasive diseases in this risk group . Moreover, promising results regarding prevention of pneumonia and acute otitis media have been published . Unfortunately, protection is raised against a limited number of pneumococcal serotypes, and serotype replacement and subsequent vaccine failure have become a serious concern . Currently, several pneumococcal surface proteins are considered as alternative vaccine candidates because of their serotype-independence . Thus far, pneumococcal surface adhesin A (PsaA) has proven to be highly protective against colonization in animal models . Moreover, pneumococcal surface protein A (PspA) and pneumolysin have shown to elicit protection against invasive diseases . Future research will elucidate their true potential in protecting humans . In this paper we discuss the present knowledge on pneumococcal vaccines and the current status of novel vaccine strategies.

Int J Med Microbiol, 2004 Apr, 293(7-8), 529 - 37
Superantigens: structure-function relationships; Baker MD et al.; Superantigens are a class of highly potent immuno-stimulatory molecules produced by Staphylococcus aureus and Streptococcus pyogenes . These toxins possess the unique ability to interact simultaneously with MHC class II molecules and T-cell receptors, forming a trimolecular complex that induces profound T-cell proliferation . The resultant massive cytokine release causes epithelial damage and leads to capillary leak and hypotension . The staphylococcal superantigens are designated staphylococcal enterotoxins A, B, C (and antigenic variants), D, E, and the recently discovered enterotoxins G to Q, and toxic shock syndrome toxin-1 . The streptococcal superantigens include the pyrogenic exotoxins A (and antigenic variants), C, G-J, SMEZ, and SSA . Superantigens are implicated in several diseases including toxic shock syndrome, scarlet fever and food poisoning; and their function appears primarily to debilitate the host sufficiently to permit the causation of disease . Structural studies over the last 10 years have provided a great deal of information regarding the complex interactions of these molecules with their receptors . This, combined with the wealth of new information from genomics initiatives, have shown that, despite their common molecular architecture, superantigens are able to crosslink MHC class II molecules and T-cell receptors by a variety of subtly different ways through the use of various structural regions within each toxin.

FEBS Lett, 2004 May 21, 566(1-3), 190 - 4
Streptococcal antigen I/II binds to extracellular proteins through intermolecular beta-sheets; Kelemen L et al.; One of the functions associated with the oral streptococcal surface protein I/II is to bind to human extracellular matrix molecules or blood components, which could act as opportunistic ligands in pathological circumstances . In order to understand the relative specificity of the binding repertoire of this bacterial adhesin, we examined by infrared measurements the mode of binding of the protein I/II from Streptococcus mutans OMZ175 (I/IIf) to fibronectin and fibrinogen . This approach revealed the beta-structure forming capacity of I/IIf upon interaction with both proteins . The forming of intermolecular beta-structures may provide a non-selective way of interaction between I/IIf and its possible targets.

J Intern Med, 2004 Jun, 255(6), 664 - 73
A prospective study on antibody response to repeated vaccinations with pneumococcal capsular polysaccharide in splenectomized individuals with special reference to Hodgkin's lymphoma; Landgren O et al.; BACKGROUND: Splenectomy is accompanied by a life-long risk of overwhelming postsplenectomy infection (OPSI), mainly caused by polysaccharide (PS) encapsulated bacteria such as Streptococcus pneumoniae . Despite extensive prophylactic efforts the mortality and morbidity rates remain high . The present study was based on a strategy with a predefined vaccination algorithm including repeated 23-valent pneumococcal vaccinations and monitoring of pneumococcal antibody levels . The antibody levels of splenectomized Hodgkin's lymphoma (HL) patients were compared with those patients splenectomized due to immune-mediated cytopenias {autoimmune haemolytic anaemia (AIHA) and immune thrombocytopenic purpura (ITP)} and also individuals who were splenectomized because of trauma (TRAUMA) . METHODS: A total of 311 splenectomized individuals were included in this prospective study (208 HL; 15 AIHA; 60 ITP; 28 TRAUMA) . Depending on their individual anti-PS antibody levels measured by enzyme-linked immunosorbent assay technique the patients were revaccinated with 23-valent pneumococcal PS vaccine up to four times in accordance with the predefined algorithm . For each vaccination occasion, serum was collected at vaccination, after 1 month +/- 2 weeks (peak), and after 1 year +/- 6 months (follow-up) . Patient files, a national population-based database, and microbiological databases were checked for 124 HL patients to identify OPSI . RESULTS: A significant response was recorded on primary vaccination as well as on two revaccination occasions for HL, AIHA/ITP, as well as TRAUMA patients . None of the variables age, gender, or time elapsed between splenectomy and first pneumococcal vaccination was found to be associated with mean PS antibody levels at prevaccination, peak or follow-up . No severe adverse events were reported . Amongst 124 clinically monitored HL patients, 10 OPSI were recorded in seven patients during the study period . One of these patients, a middle-aged female, died as a result of fulminant pneumococcal bacteraemia, which was her third OPSI during a 7-year period . CONCLUSIONS: A significant response to pneumococcal PS vaccination was found in all three groups (HL, AIHA/ITP and TRAUMA) of splenectomized patients . Importantly, both primary and repeated vaccinations were safe . Until further knowledge is gained regarding the protective concentration of serotype-specific antibody concentrations we believe that the value of vaccination and frequent revaccination (every 1-5 years) in combination with education of patients and health care professionals and clinical monitoring is beneficial for these patients at risk for OPSI.

Recenti Prog Med, 2004 Apr, 95(4), 200 - 3
{Clinical criteria to diagnosing of streptococcal pharyngitis in young adult patients}; De Socio GV et al.; Most sore throat is due to viral upper respiratory tract infections, whereas the frequency of antibiotic use in adult patients is about 73% . We evaluate the clinical guidelines (approved by the CDC) about appropriate antibiotic use for acute pharyngitis in young adults . We observe a low prevalence (3,2%) of group A beta haemolytic streptococcus (GABHS) in a population of university students . Minimizing unnecessary antimicrobial therapy in this setting is highly desirable . Use of clinical criteria (Centor algorithm) does indeed identify patients whose risk for GABHS infection is so low that microbiological testing or antibiotic treatment is unnecessary.

Bull Acad Natl Med, 2003, 187(8), 1477 - 86; discussion 1486-8
{New pneumococcal vaccine}; Gaudelus J; Streptococcus Pneumoniae is the main pathogen bacteria responsible for invasive diseases (bacteremia, meningitis) in children less than 2 years of age . The new conjugate heptavalent pneumococcal vaccine (7 polysaccharide serotypes 4, 6B, 9V, 14, 18C, 19F, 23F conjugated with the CRP 197 protein derived from diphteria anatoxin) is a great advance . This vaccine is well tolerated, immunogenic and efficient in infant . His efficacy in invasive pneumococcal diseases is more than 95% . The reduction of pneumonia (with 2.5 cm X-Ray opacity) was 32.2% in the first year of age and 23.4% in the two first years in the vaccinated group . The efficacy in acute otitis media is poor French indications of this vaccine, limited to infants under 2 years of age with further risk factor, might be extended to all the infants under 2 years of age.

An Sist Sanit Navar, 2004 Jan-Apr, 27(1), 37 - 43
{Antibiotic sensitivity and treatment recommendations for Streptococcus pneumoniae}; Gil-Setas A et al.; The aims of present paper were to determine the susceptibility of the strains to the most usual antibiotics in clinical practice and to review the current recommendations to guide the most appropriate treatment . During the period october 2000 to september 2002, the patient's data (age and sex), source of the sample, diagnosis and antibiotic susceptibility were collected on Streptococcus pneumoniae isolates from microbiology laboratories in the Navarra region (555.829 inhabitants) . Four hundred and sixty five isolates were identified (166 from invasive infections) . Generally, isolates from ear swabs were the most resistant to the antimicrobials tested, while those from blood culture were the most susceptible . Of the Streptococcus pneumoniae tested, 43% were resistant to penicillin, 6.1% to amoxicillin and 6.6% to cefotaxime . Of the 36.3% of Streptococcus pneumoniae isolates that were resistant to erythromycin, 85.45% exhibited the MLSB phenotype while the remaining 14.55% presented with the M phenotype . Multiple-resistance was detected in 32.3% of the strains . The antibiotic resistance rates to beta-lactams (specially penicillin, amoxicillin and cefotaxime/ceftrixone) in Streptococcus pneumoniae don't prevent its clinical use for the most of Streptococcus pneumoniae isolated in our area, except for pneumococcal meningitis.

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