FEBS Lett, 2002 Jul 17, 523(1-3), 239 - 46 The crystal structure of anthranilate phosphoribosyltransferase from the enterobacterium Pectobacterium carotovorum; Kim C et al.; The structure of anthranilate phosphoribosyltransferase from the enterobacterium Pectobacterium carotovorum has been solved at 2.4 A in complex with Mn(2+)-pyrophosphate, and at 1.9 A without ligands . The enzyme structure has a novel phosphoribosyltransferase (PRT) fold and displays close homology to the structures of pyrimidine nucleoside phosphorylases . The enzyme is a homodimer with a monomer of 345 residues . Each monomer consists of two subdomains, alpha and alpha/beta, which form a cleft containing the active site . The nature of the active site is inferred from the trapped MnPPi complex and detailed knowledge of the active sites of nucleoside phosphorylases . With the anthranilate (An)PRT structure solved, the structures of all the enzymes required for tryptophan biosynthesis are now known. Chin Med J (Engl), 2002 Jun, 115(6), 815 - 8 Identification of Shiga-like toxin Escherichia coli isolated from children with diarrhea by polymerase chain reaction; Zhu Q et al.; OBJECTIVE: To evaluate the etiology of Shiga like toxin producing Escherichia coli (SLTEC) in children with diarrhea . METHODS: We designed and synthesized 3 pairs of primers located in the SLT1, SLT2, and eaeA genes of enterohaemorrhagic Escherichia coli (EHEC), while the virulent genes SLT1, SLT2, and eaeA from E.coli species were amplified by polymerase chain reaction (PCR) . RESULTS: One strain of EHEC with SLT1, SLT2, and eaeA in 29 reference strains of diarrhea-causing E.coli (DCEC) and 10 strains of other enterobacteria detected by PCR had positive reactions, while all other DCEC and enterobacteria were negative . Of 474 strains of E . coli isolated from 1032 children with diarrhea and detected by PCR, 20 strains of SLT1 producing E . coli (4.2%) positive, and 7 strains of SLT2 producing E . coli (1.5%) positive; while of 74 strains of entero-SLTs-producing and invasive Escherichia coli (ESIEC), 15 strains of SLT1 (20.3%) and 5 strains of SLT2 (6.8%) were positive . CONCLUSION: Shiga-like toxin E . coli has been identified as a major etiologic agent of children with diarrhea in Taiyuan, China. Mol Microbiol, 2002 Jul, 45(2), 333 - 49 Molecular characterization of long direct repeat (LDR) sequences expressing a stable mRNA encoding for a 35-amino-acid cell-killing peptide and a cis-encoded small antisense RNA in Escherichia coli; Kawano M et al.; Genome sequence analyses of Escherichia coli K-12 revealed four copies of long repetitive elements . These sequences are designated as long direct repeat (LDR) sequences . Three of the repeats (LDR-A, -B, -C), each approximately 500 bp in length, are located as tandem repeats at 27.4 min on the genetic map . Another copy (LDR-D), 450 bp in length and nearly identical to LDR-A, -B and -C, is located at 79.7 min, a position that is directly opposite the position of LDR-A, -B and -C . In this study, we demonstrate that LDR-D encodes a 35-amino-acid peptide, LdrD, the overexpression of which causes rapid cell killing and nucleoid condensation of the host cell . Northern blot and primer extension analysis showed constitutive transcription of a stable mRNA (approximately 370 nucleotides) encoding LdrD and an unstable cis-encoded antisense RNA (approximately 60 nucleotides), which functions as a trans-acting regulator of ldrD translation . We propose that LDR encodes a toxin-antitoxin module . LDR-homologous sequences are not pre-sent on any known plasmids but are conserved in Salmonella and other enterobacterial species. Intensive Care Med, 2002 Jul, 28(7), 857 - 63 Epub 2002 May 30. Reproducibility of protected specimen brush and bronchoalveolar lavage conserved at 4 degrees C for 48 hours; Forceville X et al.; OBJECTIVE: Protected specimen brush (PSB) and bronchoalveolar lavage (BAL) are proposed in combination to optimize antimicrobial treatment . Nevertheless, they are only validated for immediate laboratory processing . This study was therefore conducted to determine whether 48 h conservation at a mere 4 degrees C enables good culture reproducibility for both PSB and BAL . DESIGN AND SETTING: Prospective study, evaluation of a conservation procedure for PSB and BAL, from February 1994 to February 1995, in the 12 bed ICU of a general hospital (938 beds) . SAMPLES: Ninety-nine PSB and 86 BAL samples, obtained from 100 bronchoscopic procedures, were analyzed . Thresholds were 10(3) and 10(4) cfu/ml for PSB and BAL, respectively . MEASUREMENTS AND RESULTS: Qualitative comparison between the immediate and 48 h procedures were, for PSB, specificity 100%, sensitivity 78%, positive predictive value 100%, negative predictive value 84% and overall accuracy 90%; and for BAL: 100%, 89%, 100%, 89% and 94% . Lowered 10(2) and 10(3) cfu/ml thresholds at the 48 h procedure for PSB and BAL reduce the false negatives from 10 to 3 and 5 to 1, respectively . Microorganism results were comparable for PSB and BAL ( r = 0.63 and 0.67), especially for the most resistant strains: Staphylococcus, Enterobacteriaceae and Pseudomonas . However, there was a decrease in the Neisseria and Haemophilus group ( p < 0.01) . CONCLUSION: There is a good culture reproducibility for both PSB and BAL after 48 h conservation at 4 degrees C, especially with lowered thresholds; this technique is therefore appropriate for routine use. Antimicrob Agents Chemother, 2002 Aug, 46(8), 2656 - 61 Novel complex sul1-type integron in Escherichia coli carrying bla(CTX-M-9); Sabate M et al.; For the present report, a novel complex class 1 integron, In60, was characterized . Part of this integron includes the bla(CTX-M-9) gene and its downstream nucleotide sequence, which shares 81% and 78% nucleotide identity with those of kluA-1 beta-lactamase and orf3 of K . ascorbata, respectively . Furthermore, a new insertion sequence, IS3000, has been found in In60 . PCR analysis indicates that integron In60 is present in 33 of 34 nonclonal enterobacterial isolates carrying the putative beta-lactamase CTX-M-9. Antimicrob Agents Chemother, 2002 Aug, 46(8), 2640 - 3 The AcrAB-TolC efflux pump contributes to multidrug resistance in the nosocomial pathogen Enterobacter aerogenes; Pradel E et al.; We identified the genes encoding the AcrA-AcrB-TolC efflux pump in Enterobacter aerogenes and constructed acrAB and tolC mutants from a multidrug-resistant isolate . Both derivatives were more susceptible to antibiotics than the parental strain . Sequence analysis and complementation experiments revealed that the multidrug-resistant isolate is an acrR mutant. Physiol Plant, 2002 Aug, 115(4), 513 - 522 Overexpression of a bacterial indole-3-acetyl-l-aspartic acid hydrolase in Arabidopsis thaliana; Tam YY et al.; Transgenic Arabidopsis lines (ecotype Col-0) carrying the Enterobacter agglomerans IaaspH gene under CaMV 35S promoter control were more sensitive to exogenous indole-3-acetyl aspartic acid (IAA-Asp) and metabolized {2'-14C}IAA-Asp more rapidly than control lines . Free IAA, total IAA and IAN levels in independent transgenic lines that accumulated IaaspH mRNA varied insignificantly from control levels, yet IAA-Asp levels were significantly reduced . The transgenic lines were grown in a variety of conditions and subjected to morphometric analysis . All three lines showed statistically significant differences in rosette diameter (in soil), root and hypocotyl length (on agar) . These effects were transient in some cases and did not manifest themselves under all growth conditions tried . The two independent lines with single T-DNA insertions had lower seed set compared to control lines. Equine Vet J, 2002 Jul, 34(4), 366 - 72 Pharmacokinetics of marbofloxacin in horses; Bousquet-Melou A et al.; Marbofloxacin is a fluoroquinolone antibiotic expected to be effective in the treatment of infections involving gram-negative and some gram-positive bacteria in horses . In order to design a rational dosage regimen for the substance in horses, the pharmacokinetic properties of marbofloxacin were investigated in 6 horses after i.v., subcutaneous and oral administration of a single dose of 2 mg/kg bwt and the minimal inhibitory concentrations (MIC) assessed for bacteria isolated from equine infectious pathologies . The clearance of marbofloxacin was mean +/- s.d . 0.25 +/- 0.05 l/kg/h and the terminal half-life 756 +/- 1.99 h . The marbofloxacin absolute bioavailabilities after subcutaneous and oral administration were 98 +/- 11% and 62 +/- 8%, respectively . The MIC required to inhibit 90% of isolates (MIC90) was 0.027 microg/ml for enterobacteriaceae and 0.21 microg/ml for Staphylococcus aureus . The values of surrogate markers of antimicrobial efficacy (AUIC, Cmax/MIC ratio, time above MIC90) were calculated and the marbofloxacin concentration profiles simulated for repeated administrations . These data were used to determine rational dosage regimens for target bacteria . Considering the breakpoint values of efficacy indices for fluoroquinolones, a marbofloxacin dosage regimen of 2 mg/kg bwt/24 h by i.v., subcutaneous or oral routes was more appropriate for enterobacteriaceae than for S . aureus. Am J Med, 2002 Jul 8, 113 Suppl 1A, 14S - 19S The etiology of urinary tract infection: traditional and emerging pathogens; Ronald A; The microbial etiology of urinary infections has been regarded as well established and reasonably consistent . Escherichia coli remains the predominant uropathogen (80%) isolated in acute community-acquired uncomplicated infections, followed by Staphylococcus saprophyticus (10% to 15%) . Klebsiella, Enterobacter, and Proteus species, and enterococci infrequently cause uncomplicated cystitis and pyelonephritis.The pathogens traditionally associated with UTI are changing many of their features, particularly because of antimicrobial resistance . The etiology of UTI is also affected by underlying host factors that complicate UTI, such as age, diabetes, spinal cord injury, or catheterization . Consequently, complicated UTI has a more diverse etiology than uncomplicated UTI, and organisms that rarely cause disease in healthy patients can cause significant disease in hosts with anatomic, metabolic, or immunologic underlying disease . The majority of community-acquired symptomatic UTIs in elderly women are caused by E coli . However, gram-positive organisms are common, and polymicrobial infections account for up to 1 in 3 infections in the elderly . In comparison, the most common organisms isolated in children with uncomplicated UTI are Enterobacteriaceae . Etiologic pathogens associated with UTI among patients with diabetes include Klebsiella spp., Group B streptococci, and Enterococcus spp., as well as E coli . Patients with spinal cord injuries commonly have E coli infections . Other common uropathogens include Pseudomonas and Proteus mirabilis.Recent advances in molecular biology may facilitate the identification of new etiologic agents for UTI . The need for accurate and updated population surveillance data is apparent, particularly in light of concerns regarding antimicrobial resistance . This information will directly affect selection of empiric therapy for UTI. Appl Microbiol Biotechnol, 2002 Jul, 59(2-3), 303 - 9 Epub 2002 May 03. Use of carbon and energy balances in the study of the anaerobic metabolism of Enterobacter aerogenes at variable starting glucose concentrations; Converti A et al.; The anaerobic metabolism of Enterobacter aerogenes was studied in batch culture at increasing initial glucose levels (9.0< S(o) <72 g l(-1)) . The ultimate concentrations of fermentation products were utilized to check a metabolic flux analysis based on simple carbon mass and energy balances that promise to be suitable for the study of different fermentation processes, either under aerobic or anaerobic conditions . The stoichiometric coefficients of products collected at increasing starting glucose concentrations under anaerobic conditions suggest: (a) little influence of starting glucose level on the formation of the main fermentation products (2,3-butanediol and ethanol); (b) possible inhibition of 2,3-butanediol and lactate formations by increased ethanol concentration; (c) consequent increase in carbon flux through the remaining metabolic pathways with increased molar productions of succinate, acetate and hydrogen; (d) relative constancy of the molar production of ATP and CO(2). J Biol Chem, 2002 Sep 13, 277(37), 33811 - 7 Epub 2002 Jul 10. The respiratory complex I (NDH I) from Klebsiella pneumoniae, a sodium pump; Gemperli AC et al.; The electrogenic NADH:Q oxidoreductase from the enterobacterium Klebsiella pneumoniae transports Na(+) ions . The complex was purified with an increase of the specific Na(+) transport activity from 0.2 micromol min(-1) mg(-1) in native membrane vesicles to 4.7 micromol min(-1) mg(-1) in reconstituted enzyme specimens . The subunit pattern resembled that of complex I from Escherichia coli, and two prominent polypeptides were identified as the NuoF and NuoG subunits of complex I . During purification the typical cofactors of complex I were enriched to yield approximately 17 nmol mg(-1) iron, 24 nmol mg(-1) acid-labile sulfide, and 0.79 nmol mg(-1) FMN in the purified sample . The enzyme contained approximately 1.2 nmol mg(-1) Q6 and 1.5 nmol mg(-1) Q8 . The reduction of ubiquinone by NADH was Na(+)-dependent, which indicates coupling of the chemical and the vectorial reaction of the pump . The Na(+) activation profile corresponded to the Hill equation with a Hill coefficient K(H)(Na(+)) = 1.96 and with a half-maximal saturation at 0.33 mm Na(+) . The reconstituted complex I from Klebsiella pneumoniae catalyzed deamino-NADH oxidation, Q1 reduction, and Na(+) translocation with specific activities of 2.6 units mg(-1), 2.4 units mg(-1), and 4.7 units mg(-1), respectively, which indicate a Na(+)/electron stoichiometry of one. Nahrung, 2002 Jun, 46(3), 197 - 9 Biogenic amines in chicken meat products in relation to bacterial load, pH value and sodium chloride content; Nassar AM et al.; Exogenous biogenic amines may present food poisoning hazards, especially when coupled with additional risk factors . In recent years, consumption of poultry meat and poultry meat products has increased . Therefore, the goals of this work were to estimate biogenic amines in chicken meat products and to find a correlation to bacterial quality, pH value and/or NaCl content . A total number of 60 random samples of chicken frankfurter, luncheon and keshta were collected from Cairo and Assiut supermarkets and groceries . The samples were analyzed for estimation of histamine, tyramine, cadaverine and puerscine . The results were in low levels and varied in each product . The bacterial quality was evaluated with respect to total colony, Enterobacteriaceae, coliforms, Lactobacillaceae and Pseudomonas aeruginosa counts . These amineproducering bacteria were detected in low amounts . Such findings as well as pH value and NaCl content correlated with amines levels. J Bacteriol, 2002 Aug, 184(15), 4205 - 10 Cooperativity in signal transfer through the Uhp system of Escherichia coli; Verhamme DT et al.; The UhpABC regulatory system in enterobacteria controls the expression of the hexose phosphate transporter UhpT . Signaling is initiated through sensing of extracellular glucose 6-phosphate by membrane-bound UhpC, which in turn modulates the histidine-protein kinase UhpB . Together with the cytoplasmic response regulator UhpA, they constitute a typical two-component regulatory system based on His-to-Asp phosphoryl transfer . Activated (i.e., phosphorylated) UhpA binds to the promoter region of uhpT, resulting in initiation of transcription . We have investigated the contribution of transmembrane signaling (through UhpBC) and intracellular activation (through UhpA) to the overall Uhp response (UhpT expression) in vivo . UhpA activation could be made independent of transmembrane signaling when (Delta)uhpBC cells were grown on pyruvate . Inorganic phosphate interfered with glucose 6-phosphate-dependent, UhpBC-mediated, as well as pyruvate-mediated activation of UhpA . The relationship between the concentration of inducer (glucose 6-phosphate) and the Uhp induction rate was nonhyperbolic, indicating positive cooperativity . The degree of cooperativity was affected by the carbon or energy source available to the cells for growth . As pyruvate-mediated activation of UhpA in (Delta)uhpBC cells could result in considerably stronger UhpT expression than glucose 6-phosphate-dependent activation through UhpBC, the observed positive cooperativity for the overall pathway in wild-type cells may reflect the previously described cooperative binding of UhpA to the uhpT promoter (J . L . Dahl et al., J . Biol . Chem . 272:1910-1919, 1997). Lett Appl Microbiol, 2002, 35(2), 136 - 40 Dietary influence of kefir on microbial activities in the mouse bowel; Marquina D et al.; AIMS: In this work the microflora present in kefir, a fermented milk product, was studied together with the effect of kefir administration on different groups of indigenous bacteria of mouse bowel . METHODS AND RESULTS: Kefir microflora was composed of lactic acid bacteria, acetic acid bacteria and yeasts . Yeast population was composed of Saccharomyces cerevisiae, S . unisporus, Candida kefir, Kluyveromyces marxianus and K . lactis . The streptococci levels in kefir treated mice increased by 10-fold and the levels of sulfite-reducing clostridia decreased by 100-fold . The number of lactic acid bacteria increased significantly . CONCLUSIONS: The administration of kefir significantly increased the lactic acid bacteria counts in the mucosa of the bowel . Ingestion of kefir specifically lowered microbial populations of Enterobacteriaceae and clostridia . SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first long-term study about the effects of the kefir administration on the intestinal microflora of mice. Microbiol Res, 1995 Sep, 150(3), 305 - 9 Survival of enterobacteria in liquid cultures during microwave radiation and conventional heating; Papadopoulou C et al.; Bacteria in food have been reported to survive in larger numbers after processing by microwave radiation than after conventional processing . The bactericidal effect of a domestic microwave oven (SHARP R-7280) on certain pathogenic enterobacteria species was investigated in vitro, in comparison with conventional heating (boiling) . The death rates of different nosocomial strains of Escherichia coli, Salmonella sofia, Salmonella enteritidis, Proteus mirabilis and Pseudomonas aeruginosa were tested . The microwave oven and the conventional heating system used were both calibrated in order to calculate temperatures from exposure times . For each strain duplicate samples of 25 ml of pure culture with concentrations at least 10(6) cfu/ml were exposed to microwave radiation . An equal number of samples of the same volume and concentration were exposed to conventional heating . Subsequently all samples were examined qualitatively and quantitatively following standard microbiological procedures . The results indicate that microwaves have an efficient bactericidal effect on the enterobacteria in liquid cultures. BMC Microbiol . 2002 Jun 26;2(1):15. Detection of Mycobacterium avium subspecies paratuberculosis specific IS900 insertion sequences in bulk-tank milk samples obtained from different regions throughout Switzerland; Corti S et al.; BACKGROUND: Since Mycobacterium avium subspecies paratuberculosis (MAP) was isolated from intestinal tissue of a human patient suffering Crohn's disease, a controversial discussion exists whether MAP have a role in the etiology of Crohn's disease or not . Raw milk may be a potential vehicle for the transmission of MAP to human population . In a previous paper, we have demonstrated that MAP are found in raw milk samples obtained from a defined region in Switzerland . The aim of this work is to collect data about the prevalence of MAP specific IS900 insertion sequence in bulk-tank milk samples in different regions of Switzerland . Furthermore, we examined eventual correlation between the presence of MAP and the somatic cell counts, the total colony counts and the presence of Enterobacteriaceae . RESULTS: 273 (19.7%) of the 1384 examined bulk-tank milk samples tested IS900 PCR-positive . The prevalence, however, in the different regions of Switzerland shows significant differences and ranged from 1.7% to 49.2% . Furthermore, there were no statistically significant (p >> 0.05) differences between the somatic cell counts and the total colony counts of PCR-positive and PCR-negative milk samples . Enterobacteriaceae occur as often in IS900 PCR-positive as in PCR-negative milk samples . CONCLUSION: This is the first study, which investigates the prevalence of MAP in bulk-tank milk samples all over Switzerland and infers the herd-level prevalence of MAP infection in dairy herds . The prevalence of 19.7% IS900 PCR-positive bulk-milk samples shows a wide distribution of subclinical MAP-infections in dairy stock in Switzerland . MAP can therefore often be transmitted to humans by raw milk consumption. Biosci Biotechnol Biochem, 2002 May, 66(5), 1143 - 7 Functional expression of chitinase and chitosanase, and their effects on morphologies in the yeast Schizosaccharomyces pombe; Shimono K et al.; The chitinase A (chiA) gene from Enterobacter sp . G-1 and the chitosanase A (choA) gene from Matsuebacter chitosanotabidus 3001 were expressed separately and simultaneously in the yeast Schizosaccharomyces pombe . The chiA gene was placed under the transcriptional control of the nmt1 promoter with the glutathione S transferase (GST), and the choA gene was expressed by the human cytomegalovirus (hCMV) promoter with the P factor secretion signal (P3) . The expressed proteins of ChoA and GST-ChiA were enzymatical active and were detected as 34-kDa and 80-kDa, respectively, by Western blot analysis . The transformant that expressed the choA gene was able to secrete ChoA efficiently into the culture medium with the specific activity of 102.36 U/mg protein . When the chiA gene was expressed in S . pombe, yeast cells grew slowly and cells became elongated, but when the choA gene was expressed, cells became swollen . Expression of both the chiA and the choA genes resulted in elongated and fat cells. J Food Prot, 2002 Jun, 65(6), 981 - 7 Combined effect of gamma irradiation, ascorbic acid, and edible coating on the improvement of microbial and biochemical characteristics of ground beef; Ouattara B et al.; This study was conducted to evaluate the combined effect of gamma irradiation and the incorporation of natural antimicrobial compounds in cross-linked films on the microbiological and biochemical characteristics of ground beef . Medium-fat (23% fat) ground beef patties were divided into three separate treatment groups: (i) control samples without additives, (ii) ground beef samples containing 0.5% (wt/wt) ascorbic acid, and (iii) ground beef samples containing 0.5% ascorbic acid and coated with a protein-based cross-linked film containing immobilized spice powders . Meat samples were irradiated at doses of 0, 1, 2, and 3 kGy and stored at 4 +/- 2 degrees C . Microbial growth (based on total aerobic plate counts {APCs} and total coliforms) was evaluated, as were the content of thiobarbituric acid-reactive substances (TBARS) and that of free sulfydryl groups . At the end of the storage period, Enterobacteriaceae, presumptive Staphylococcus aureus, presumptive Pseudomonas spp., Brochothrix thermosphacta, and lactic acid bacteria were enumerated . Regardless of the treatment group, irradiation significantly (P < or = 0.05) reduced the APCs . Irradiation doses of 1, 2, and 3 kGy produced immediate APC reductions of 2, 3, and 4 log units, respectively . An APC level of 6 log CFU/g was reached after 4, 7, and 10 days for samples irradiated at 1, 2, and 3 kGy, respectively . Lactic acid bacteria and B . thermosphacta were more resistant to irradiation than were Enterobacteriaceae and Pseudomonas . The TBARS and free sulfydryl contents were stabilized during postirradiation storage for samples containing ascorbic acid and coated with the protein-based cross-linked film containing immobilized spice powders. Rev Med Chir Soc Med Nat Iasi, 2001 Jan-Mar, 105(1), 137 - 41 {Detection of extended-spectrum beta-lactamases production in enterobacteriaceae strains}; Tuchilus C et al.; Extended-spectrum beta-lactamases (ESBL) cause resistance to ceftazidime, ceftriaxone, cefotaxime and other cephalosporins . The ESBL enzymes are most often derivatives of beta-lactamases from the TEM or SHV group . The aim of our work was to study ESBL production by 35 ceftazidime-resistant strains . We selected these strains among 335 Enterobacteriaceae, isolated in Eastern Romania from healthy carriers and patients, between january-june 1999 . To identify the studied strains we used API 20E and API 32E . The ESBL production was established by using both double disk synergy test (DDT) and EXPERT computer program of miniAPI (Bio Merieux) . We revealed ESBL production in 26 strains when EXPERT program was used and only 23 when DDT was used . Our results suggest that miniAPI is a better method to study ESBL production than DDT. Rev Med Chir Soc Med Nat Iasi, 2000 Apr-Jun, 104(2), 131 - 4 Sensitivity of Enterobacteriaceae strains to the third generation cephalosporins; Poiata A et al.; The aim of this study was to evaluate the in vitro activity of cefotaxime, ceftriaxone, ceftazidime, ceftibuten and aztreonam against the clinical isolates of Enterobacteriaceae . MICROORGANISMS: A total of 973 enterobacteria isolated from clinical material (urine and faeces) were tested . Strains were collected during 1996-1998 period, in some centers of Eastern Romania . SENSITIVITY STUDY: MICs were performed using the agar dilution method following standardized techniques (NCCLS, 1994) . Stock solutions of antibacterial agents were prepared from their respective powder forms, in concentrations ranging from 0.03 to 128 mg/ml . Our results show a good activity of the tested antimicrobials on Escherichia coli, Shigella spp and Klebsiella spp tested, and reveal a particular behaviour for Salmonella spp, characterized by a heterogenous susceptibility . In conclusion, cefotaxime, ceftriaxone, ceftazidime, ceftibuten and aztreonam were active at low concentrations against tested isolates . Against Salmonella spp, these agents were less active: percent of resistant strains by beta-lactamase production was significant. West Indian Med J, 2002 Mar, 51(1), 21 - 4 Nosocomial infections in an intensive care unit in a private hospital; Orrett FA; The nosocomial infection rate in the intensive care unit (ICU) of a private hospital was assessed during an 18-month survey . From 629 admissions to the ICU, 139 hospital-acquired infections were identified . The rate was 22.1% compared to the overall nosocomial infection rate of 15.3% for the entire hospital . In the ICU, the main infections occurred in the respiratory tract, 41 (29.5%), followed by surgical wounds, 35 (25.2%), urinary tract, 28 (20.1%) and the blood stream, 24 (17.3%) . From 165 bacterial isolates, 80% of isolates were gram-negative rods, with P aeruginosa, 48 (36.6%), being the predominant gram-negative isolate followed by Klebsiella pneumoniae, 27 (20.6%), and Enterobacter sp, 22 (16.8%) . The main gram-positive isolates were S aureus, 23 (41.8%), coagulase-negative Staphylococci, 17 (30.9%), and Enterococci, 11 (20.0%) . Of the 23 S aureus strains, 15 (65.2%) were methicillin-resistant (MRSA), (8 MRSA were from surgical wounds, 5 from the respiratory tract and 2 from infected urine) . Only 2 of the 17 (11.8%) coagulase-negative staphylococci were methicillin-resistant, and both were isolated from wounds . Resistance to ampicillin and augmentin (amoxicillin-clavulanic acid) was high, 81.9% and 55.4%, respectively . Gentamicin, aztreonam, piperacillin and piperacillin-tazobactam showed resistance rates of less than 15% . Infection control measures aimed at reducing nosocomial infections at the hospital are often frustrated by apathy of hospital administrators who apparently are insensitive to the high nosocomial infection rate . Effort by the infection control team through seminars, lectures and newsletters have begun to show improvements in attitude and awareness of staff to infection control and preventative measures within the institution. J Infect Dis, 2002 Jul 1, 186(1), 49 - 56 Epub 2002 Jun 10. Evidence of extensive interspecies transfer of integron-mediated antimicrobial resistance genes among multidrug-resistant Enterobacteriaceae in a clinical setting; Leverstein-van Hall MA et al.; Multidrug resistance in gram-negative bacteria appears to be primarily the result of the acquisition of resistance genes by horizontal transfer . To what extent horizontal transfer may be responsible for the emergence of multidrug resistance in a clinical setting, however, has rarely been investigated . Therefore, the integron contents of isolates collected during a nosocomial outbreak of genotypically unrelated multidrug-resistant Enterobacteriaceae were characterized . The integron was chosen as a marker of transfer because of its association with multiresistance . Some genotypically identical isolates harbored different integrons . Grouping patients carrying the same integron yielded 6 epidemiologically linked clusters, with each cluster representing a different integron . Several patients carried multiple species harboring the same integron . Conjugation experiments with these strains resulted in the transfer of complete resistance patterns at high frequencies (10(-2) to 10(-4)) . These findings provide strong evidence that the horizontal transfer of resistance genes contributed largely to the emergence of multidrug-resistant Enterobacteriaceae in this clinical setting. Appl Environ Microbiol, 2002 Jul, 68(7), 3588 - 96 Site-specific recombination-based genetic system for reporting transient or low-level gene expression; Casavant NC et al.; We report here the construction, characterization, and application of a plasmid-based genetic system that reports the expression of a target promoter by effecting an irreversible, heritable change in a bacterial cell . This system confers strong repression of the reporter gene gfp in the absence of target promoter expression and utilizes the site-specific recombination machinery of bacteriophage P22 to trigger high-level reporter gene expression in the original cell and its progeny after target gene induction . We demonstrate the effectiveness of this genetic system by tailoring it to indicate the availability of arabinose to the biological control agent Enterobacter cloacae JL1157 in culture and in the barley rhizosphere . The presence of bioavailable arabinose triggered the production of P22 excisionase and integrase from the reporter plasmid pAraLHB in JL1157, and this led to excision of the cI repressor gene, which is flanked by att sites, and the subsequent irreversible expression of gfp in the original cell and in its progeny . In culture, nearly 100% of an E . cloacae JL1157(pAraLHB) population expressed gfp after exposure to 6.5 to 65 microM arabinose for 3 h . We used this biosensor to demonstrate that arabinose was released from the seeds of several legumes and grass species during germination and from roots of barley seedlings grown hydroponically or in soil . When introduced into microcosms containing barley, the biosensor permitted the localization of arabinose along the roots . Arabinose was present near the root-seed junction and on the seminal roots but was not detected at the root tips . This recombination-based reporter system should be useful for monitoring bacterial exposure to transient or low levels of specific molecules directly in the environment. Appl Environ Microbiol, 2002 Jul, 68(7), 3328 - 38 Plant-dependent genotypic and phenotypic diversity of antagonistic rhizobacteria isolated from different Verticillium host plants; Berg G et al.; To study the effect of plant species on the abundance and diversity of bacterial antagonists, the abundance, the phenotypic diversity, and the genotypic diversity of rhizobacteria isolated from potato, oilseed rape, and strawberry and from bulk soil which showed antagonistic activity towards the soilborne pathogen Verticillium dahliae Kleb . were analyzed . Rhizosphere and soil samples were taken five times over two growing seasons in 1998 and 1999 from a randomized field trial . Bacterial isolates were obtained after plating on R2A (Difco, Detroit, Mich.) or enrichment in microtiter plates containing high-molecular-weight substrates followed by plating on R2A . A total of 5,854 bacteria isolated from the rhizosphere of strawberry, potato, or oilseed rape or bulk soil from fallow were screened by dual testing for in vitro antagonism towards VERTICILLIUM: The proportion of isolates with antagonistic activity was highest for strawberry rhizosphere (9.5%), followed by oilseed rape (6.3%), potato (3.7%), and soil (3.3%) . The 331 Verticillium antagonists were identified by their fatty acid methyl ester profiles . They were characterized by testing their in vitro antagonism against other pathogenic fungi; their glucanolytic, chitinolytic, and proteolytic activities; and their BOX-PCR fingerprints . The abundance and composition of Verticillium antagonists was plant species dependent . A rather high proportion of antagonists from the strawberry rhizosphere was identified as Pseudomonas putida B (69%), while antagonists belonging to the Enterobacteriaceae (Serratia spp., Pantoea agglomerans) were mainly isolated from the rhizosphere of oilseed rape . For P . putida A and B plant-specific genotypes were observed, suggesting that these bacteria were specifically enriched in each rhizosphere. Diagn Microbiol Infect Dis, 2002 Jun, 43(2), 141 - 7 Emerging fluoroquinolone-resistance for common clinically important gram-negative bacteria in Taiwan; Sheng WH et al.; The in vitro antimicrobial activity of the three most commonly used fluoroquinolones (norfloxacin, ofloxacin and ciprofloxacin) was measured for 2235 isolates of seven common pathogenic gram-negative bacteria . Collected from a major teaching hospital in Taiwan during the years 1985-86, 1989-90, and 1996-97, the samples were evaluated using the agar dilution method . The overall susceptibility to fluoroquinolones has decreased rapidly after the wide use of these antimicrobial agents . Isolates of Escherichia coli and Morganella morganii proved quite susceptible to the fluoroquinolones until 1996 . By 1996-97, 20% of the bacteria had become resistant to the fluoroquinolones . Activity against Klebsiella pneumoniae and Proteus mirabilis remained excellent, however, with more than 90% of the isolates susceptible during the survey period . Interestingly, some strains of Serratia marcescens and Pseudomonas aeruginosa were demonstrating in vitro resistance even before the fluoroquinolones were launched in Taiwan . This resistance increased rapidly, with around 20-30% of S . marcescens isolates fluoroquinolone-resistant by 1996-1997 . After the introduction of the fluoroquinolones, resistance was demonstrated for about 15% of the P . aeruginosa isolates, and 20% of Enterobacter cloacae isolates, with the susceptibility rate remaining steady state through the 1990s . Different pulsed-field gel electrophoresis patterns have been demonstrated for most fluoroquinolone-resistant isolates, suggesting that the increased resistance was not due to the spread of a single clone . We conclude a trend of increasing fluoroquinolone resistance for many common gram-negative bacteria in Taiwan, especially in recent years . Appropriate use of the new fluoroquinolones should be encouraged in order to prevent the rapid emergence and increase of fluoroquinolone-resistant bacteria. Diagn Microbiol Infect Dis, 2002 Jun, 43(2), 129 - 33 Comparative in vitro activities of ertapenem against aerobic and facultative bacterial pathogens from patients with complicated skin and skin structure infections; Pelak BA et al.; This study compared the in vitro activities of ertapenem (Merck & Co., Inc.), ceftriaxone, amoxicillin-clavulanate, and piperacillin-tazobactam against 518 aerobic and facultative bacterial pathogens isolated from 340 patients with complicated skin and skin structure infections . Ciprofloxacin was also tested against Gram-negative isolates . Gram-positive cocci accounted for 68.1% of the aerobic bacteria; Staphylococcus aureus was the most common isolate (45.6%) . The ertapenem MIC was < or = 2 microg/ml for 80.9% of isolates and > or = 8 microg/ml for 16.2% (including isolates of enterococci, methicillin-resistant S . aureus, Pseudomonas aeruginosa, and other nonfermentative Gram-negative bacteria) . Against methicillin-susceptible S . aureus, ertapenem had the most potent activity . Ertapenem was the most active drug against Enterobacteriaceae (100% susceptible), whereas amoxicillin-clavulanate was least active (66% susceptible) . Piperacillin-tazobactam was the most active drug against P . aeruginosa (100% susceptible), followed by ciprofloxacin (87% susceptible) . In summary, ertapenem was highly active in vitro against many aerobic and facultative bacterial pathogens commonly recovered from patients with complicated skin and skin structure infections. Cutis, 2002 Apr, 69(4), 261 - 4 Pyoderma faciale: gram-negative recovery by means of needle aspiration; Sudy E; Pyoderma faciale is an inflammatory disease that until now was believed to occur only in women . Bacterial pathogens have not been found in most cases, and those found included only gram-positive microorganisms . We present 2 cases consistent with pyoderma faciale--one of them a 17-year-old adolescent boy--in which gram-negative bacteria were found . Enterobacter cloacae was found in one patient, and Klebsiella oxytoca was found in the other . Needle aspiration is the technique of choice in looking for pathogens in this disease. Biophys J, 2002 Jul, 83(1), 322 - 33 Biophysical characterization of triacyl monosaccharide lipid a partial structures in relation to bioactivity; Brandenburg K et al.; Synthetic triacyl glucosamine monosaccharide lipid A part structures corresponding to the non-reducing moiety of enterobacterial lipid A with an acyloxyacyl chain linked to position 3 of the glucosamine and an unbranched chain linked to position 2 (group 1) and vice versa (group 2) were analyzed biophysically: Fourier-transform infrared spectroscopy was performed to characterize the gel-to-liquid crystalline phase transition, the phosphate band contour, and the orientation of the glucosamine with respect to the membrane surface . Small-angle x-ray diffraction was applied for the elucidation of the supramolecular aggregate structure and, with that, of the molecular shape . With fluorescence resonance energy transfer the lipopolysaccharide-binding protein (LBP)-mediated intercalation of the lipid A partial structures into phospholipid liposomes was monitored . The physical data clearly exhibit a classification of the synthetic compounds into two groups: group 1 compounds have sharp phase transitions, indicating dense acyl chain packing and an inclination of the glucosamine backbone with respect to the membrane surface of 30 degrees with the phosphate buried in the membrane . Group 2 compounds have a very broad phase transition, indicating poorly packed acyl chains, and an inclination of -30 degrees with the phosphate group sticking outward . For the first group unilamellar phases are observed superimposed by a non-lamellar structure, and for the second one only multilamellar aggregate structures . The cytokine-inducing capacity in human mononuclear cells is relatively high for the first group and low or absent for the second group . Based on these data a model of the intra and intermolecular conformations is proposed which also extends the concept of "endotoxic conformation." J Antimicrob Chemother, 2002 Jul, 50 Suppl, 1 - 5 Global antibiotic resistance in Streptococcus pneumoniae; Adam D; The last two decades of the 20th century were marked by an increasing resistance rate among several bacteria . Threat of resistance is present in Staphylococcus spp., Enterococcus spp., Pseudomonas spp . and Enterobacteriaceae, which are the major pathogens in nosocomial infections . In the community, too, increasing resistance can be observed and is attributed mainly (but not exclusively) to Streptococcus pneumoniae and Haemophilus influenzae . To scrutinize this trend, resistance surveillance in the community was established about 10 years ago . One of the multinational, longitudinal surveillance programmes in place is the Alexander Project, which was established in 1992 to monitor the susceptibility of the major community-acquired lower respiratory tract pathogens to a range of antibacterial drugs . The Alexander Project has revealed a tendency towards increasing resistance of S . pneumoniae to penicillin and macrolide therapy . Within Europe, the prevalence of penicillin resistance among S . pneumoniae isolates is particularly high in France and Spain . Macrolide resistance in S . pneumoniae is also a growing problem in European countries such as France, Spain, Belgium and Italy, where the extent of macrolide resistance in S . pneumoniae now exceeds that of penicillin resistance. Ann Intern Med, 2002 Jun 18, 136(12), 896 - 907 Update on spondyloarthropathies; Khan MA; Spondyloarthropathies are a cluster of interrelated and overlapping chronic inflammatory rheumatic diseases that primarily include ankylosing spondylitis, reactive arthritis, and the arthritis associated with psoriasis and inflammatory bowel diseases . The primary pathologic sites are the entheses (the sites of bony insertion of ligaments and tendons); the axial skeleton, including the sacroiliac joints; the limb joints; and some nonarticular structures, such as the gut, skin, eye, and aortic valve . Although spondyloarthropathies are not associated with rheumatoid factor, they show a strong association with HLA-B27; however, this association varies markedly among various spondyloarthropathies and among ethnic groups . The most widely used classification criterion, from the European Spondyloarthropathy Study Group, encompasses the currently recognized wider disease spectrum, with a sensitivity and specificity that generally exceed 85% . Spondyloarthropathies occur in genetically predisposed persons and are triggered by environmental factors, but the cellular and molecular mechanisms of inflammation are not yet fully understood . Chlamydial and many enterobacterial infections can trigger reactive arthritis, but an infectious trigger for ankylosing spondylitis has not yet been established . HLA-B27 itself is involved in enhancing genetic susceptibility, but the underlying molecular basis is still unknown; additional genes include the putative susceptibility genes for Crohn disease, ulcerative colitis, and psoriasis . A specific susceptibility gene for Crohn disease, NOD2, is located on chromosome 16q12, and one of the candidate genes for psoriasis, PSORS1, has been mapped to a 60-kb fragment on chromosome 6p, which is telomeric to the HLA-C locus . This paper reviews the efficacy of anti-tumor necrosis factor-alpha therapy and other therapeutic advances. J Appl Microbiol, 2002, 93(1), 169 - 77 Cross-contamination of carcasses and equipment during pork processing; Warriner K et al.; AIMS: The cross-contamination events within a commercial pork processing line were examined by a combination of ERIC-PCR DNA fingerprinting of Escherichia coli and plate counts . METHODS AND RESULTS: Sponge sampling of environmental surfaces and carcasses was performed over an 8-h processing period . Prior to the start of processing the scraper and dry polisher blades were found to harbour substantial Enterobacteriaceae and Escherichia coli populations . From plate count data the key cross-contamination site for the transfer of bacteria between carcasses occurred during evisceration . However, DNA fingerprints of representative E . coli isolates identified that genotypes initially present on the scraper/dry polisher became distributed on wet polisher blades, band-saw and butcher's hands despite a singeing step being performed post dry polishing . A high proportion of E . coli on post-eviscerated carcasses could be traced to down-stream (pre-singe) environmental contact surfaces . CONCLUSIONS: DNA fingerprinting has demonstrated that E . coli and potential enteric pathogens can be transferred between pork carcasses throughout the processing line . In this respect scalding and singeing cannot be relied upon to control cross-contamination of enteric bacteria between carcasses . SIGNIFICANCE AND IMPACT OF THE STUDY: Sole reliance on indicator organism counts to identify cross-contamination events as currently advocated is limited. J Appl Microbiol, 2002, 93(1), 60 - 8 Oligonucleotide probe for detecting Enterobacteriaceae by in situ hybridization; Ootsubo M et al.; AIMS: To develop oligonucleotide probes for visualizing bacteria belonging to Enterobacteriaceae . METHODS AND RESULTS: 24-mer oligonucleotide probe (probe D) was designed by comparison of 16S rDNA sequences of 35 species of Enterobacteriaceae, eight species of Vibrionaceae and six species of Pasteurellaceae . The sequence of the probe corresponding to the complementary sequence of a position 1251-1274 of Escherichia coli 16S rRNA was found to be a highly conserved region of 16S rDNA sequence in Enterobacteriaceae different from that of Vibrionaceae and Pasteurellaceae . The fluorescent dye-labelled probe was tested for the specificity by in situ hybridization and epifluorescence microscopy . Seventy-six out of 78 strains belonging to Enterobacteriaceae were visualized in an optimal hybridization condition . No bacterial strains belonging to Vibrionaceae (31 strains) and Gram-positive bacteria (three strains) were visualized . CONCLUSIONS: In situ hybridization using probe D allows the detection of bacterial cells belonging to Enterobacteriaceae without false positive reaction . SIGNIFICANCE AND IMPACT OF THE STUDY: In situ hybridization techniques using the probe D are potential tools for detecting Enterobacteriaceae in food and water samples. Med Mycol, 2002 Apr, 40(2), 131 - 7 CAY-I, a fungicidal saponin from Capsicum sp . fruit; De Lucca AJ et al.; Saponins are steroidal or terpenoid-based glycosides with surface active properties . A steroidal saponin, CAY-1, with a molecular weight of 1243.35 Da, was isolated and purified to homogeneity from commercially available dry, ground fruit of Capsicum frutescens . CAY-1 was shown to be a potent fungicide for the germinating conidia of Aspergillus flavus, A . fumigatus, A . parasiticus and A . niger with species-dependent LD90 values between 3 and 20 microM . Activity against some Aspergillus species was affected by the test medium used . In vitro assays, CAY-1 was effective against Pneumocystis carinii (IC50): 9.5 microM) and Candida albicans (IC90: 6.2 microM) . CAY-1 had no effect on the viability of the nongerminating conidia of the two filamentous fungi, P . carinii and C . albicans, nor on the conidial type of Fusarium oxysporum . It was ineffective against the bacteria Enterobacter agglomerans, Bacillus subtilis, Escherichia coli and Staphylococcus aureus . CAY-1 was not cytotoxic to A 549 lung carcinoma cells or HeLa cells at effective fungicidal concentrations . The results indicate that CAY-1 is an effective fungicide for Aspergillus species, C . albicans and P . carinii at concentrations below the threshold for mammalian cell toxicity. J Bacteriol, 2002 Jul, 184(13), 3549 - 59 Differential regulation of multiple proteins of Escherichia coli and Salmonella enterica serovar Typhimurium by the transcriptional regulator SlyA; Spory A et al.; SlyA is a transcriptional regulator of Escherichia coli, Salmonella enterica, and other bacteria belonging to the ENTEROBACTERIACEAE: The SlyA protein has been shown to be involved in the virulence of S . enterica serovar Typhimurium, but its role in E . coli is unclear . In this study, we employed the proteome technology to analyze the SlyA regulons of enteroinvasive E . coli (EIEC) and Salmonella serovar Typhimurium . In both cases, comparative analysis of the two-dimensional protein maps of a wild-type strain, a SlyA-overproducing derivative, and a corresponding slyA mutant revealed numerous proteins whose expression appeared to be either positively or negatively controlled by SlyA . Twenty of the putative SlyA-induced proteins and 13 of the putative SlyA-repressed proteins of the tested EIEC strain were identified by mass spectrometry . The former proteins included several molecular chaperones (GroEL, GroES, DnaK, GrpE, and CbpA), proteins involved in acid resistance (HdeA, HdeB, and GadA), the "starvation lipoprotein" (Slp), cytolysin ClyA (HlyE or SheA), and several enzymes involved in metabolic pathways, whereas most of the latter proteins proved to be biosynthetic enzymes . Consistently, the resistance of the EIEC slyA mutant to heat and acid stress was impaired compared to that of the wild-type strain . Furthermore, the implication of SlyA in the regulation of several of the identified E . coli proteins was confirmed at the level of transcription with lacZ fusions . Twenty-three of the Salmonella serovar Typhimurium proteins found to be affected by SlyA were also identified by mass spectrometry . With the exception of GroEL these differed from those identified in the EIEC strain and included proteins involved in various processes . The data suggest that gene regulation by SlyA might be crucial for intracellular survival and/or replication of both EIEC and Salmonella serovar Typhimurium in phagocytic host cells. Biochem Biophys Res Commun, 2002 May 24, 293(5), 1370 - 3 Inhibitors of antibiotic efflux pump in resistant Enterobacter aerogenes strains; Mallea M et al.; Enterobacter aerogenes, a nosocomial pathogen, is frequently exhibiting multidrug resistance mechanisms associated with a change in membrane permeability . In clinical isolates, active efflux plays a prominent role in antibiotic resistance . We report here the effect of three unrelated compounds that are able to restore a noticeable antibiotic susceptibility to resistant strains . The targeting of various parameters which contribute to the efficacy of the efflux mechanism, such as energy, flux selectivity, or functional assembly of the membrane complex, increases the intracellular chloramphenicol concentration in resistant isolates. Diagn Microbiol Infect Dis, 2002 May, 43(1), 61 - 4 Use of surrogate antimicrobial agents to predict susceptibility to ertapenem; Friedland IR et al.; Broth or agar dilution susceptibility test results for Enterobacteriaceae (11,775 strains), anaerobes (2888 strains), staphylococci (2206 strains), Haemophilus spp . (840 strains), group A streptococci (280 strains), group B streptococci (269 strains), Streptococcus pneumoniae (709 strains), and 160 other streptococci were analyzed to identify surrogate antimicrobial agents to predict susceptibility to ertapenem . Ertapenem MIC interpretive categories approved by the United States FDA were compared to those of imipenem, oxacillin (staphylococci), or penicillin (streptococci) . Ertapenem resistance was rare (1.2%) among 8187 consecutively collected clinical isolates of Enterobacteriaceae, including a large proportion of isolates from intensive care units . Absolute categorical agreement between ertapenem and imipenem, and very major (false susceptible) and major errors (false resistant) using imipenem to predict ertapenem results were 97.2%, 0.9%, and 0.4%, respectively, for Enterobacteriaceae (10,992 strains tested against both drugs) and 99.0%, 0.2%, and 0% for anaerobes . All Haemophilus spp., groups A and B streptococci, penicillin-susceptible and -intermediate S . pneumoniae, and other penicillin-susceptible streptococci were susceptible to ertapenem . All oxacillin-susceptible Staphylococcus aureus were ertapenem susceptible, except 1 that was intermediate . Surrogate antimicrobial agents that can be used to reliably predict ertapenem susceptibility by MIC tests are imipenem for Enterobacteriaceae and anaerobes, oxacillin for staphylococci, and penicillin for streptococci. FEMS Microbiol Lett, 2002 May 21, 211(1), 51 - 5 ERIC- and REP-PCR amplify non-repetitive fragments from the genome of Drechslera avenae and Stemphylium solani; Mehta A et al.; In the present study, the homology relationships among fragments amplified by ERIC (Enterobacterial Repetitive Intergenic Consensus) and REP (Repetitive Extragenic Palindromic) primers in Drechslera avenae and Stemphylium solani causing leaf blotch and black spot of oats and leaf blight of cotton, respectively, were investigated by Southern hybridization followed by DNA sequencing . ERIC- and REP-PCR were performed for 24 isolates of D . avenae and 33 isolates of S . solani . The banding patterns obtained were transferred to nylon membranes and a total of 16 fragments, four ERIC and four REP bands from each fungus, used as probes . The hybridization and DNA sequencing revealed a low degree of similarity among the bands indicating that the regions amplified by these primers are not repetitive and that the primers anneal at random regions in the genomes of D . avenae and S . solani. Clin Exp Rheumatol, 2002 Mar-Apr, 20(2), 193 - 200 Mononuclear cell response to enterobacteria and Gram-positive cell walls of normal intestinal microbiota in early rheumatoid arthritis and other inflammatory arthritides; Chen T et al.; OBJECTIVE: To study whether enterobacteria and Gram-positive bacterial cell walls (BCW) derivedfrom normal intestinal microbiota are involved in the etiopathogenesis of early rheumatoid arthritis (RA) . METHODS: Peripheral blood mononuclear cells (PBMC) and synovial fluid mononuclear cells (SFMC) were isolatedfrom patients with early RA (the average duration of 5 months) and the controls (other types of inflammatory arthritis) . The mononuclear cell proliferation and tumor necrosis factor-alpha (TNF-alpha) responses to heat-killed Salmonella enteritidis (SE) . Yersinia enterocolitica (YE), and Escherichia coli (EC), and to Gram-positive BCW derived from four common intestinal indigenous bacteria, Eubacterium aerofaciens (EA), Eubacterium limosum (EL), Lactobacillus casei (LC), and Lactobacillus fermentum (LF), and a BCW derived from a pathogen, Streptococcus pyogenes (SP) were investigated . RESULTS: 39% or 56% of patients with early RA showed significant proliferation responses by PBMC or SFMC against enterobacteria, respectively . In other types of arthritis, corresponding figures were 59% or 66% . When BCW were used as antigens, 8.1% or 23% of patients with early RA showed proliferation responses by PBMC or SFMC, respectively . In other types of arthritis the corresponding figures were 7.5% or 35%, respectively . However, TNF-alpha production by SFMC stimulated by EA BCW, SE, YE or EC, was significantly higher in early RA than in other types of arthritis . CONCLUSION: These results suggest that SFMC reacting with enterobacteria or BCW exist in some patients with early RA, but also in other types of inflammatory arthritis . Intestinal bacterial agents may play a role in the etiopathogenesis of RA, but the effect appears to be non-specific. Clin Microbiol Infect, 2002 Apr, 8(4), 207 - 13 Susceptibility of Enterobacteriaceae to beta-lactam agents and fluoroquinolones: a 3-year survey in France; Sirot J et al.; OBJECTIVE: To assess trends in the susceptibility to beta-lactam agents and to fluoroquinolones of clinically relevant Enterobacteriaceae isolated over a 3-year period in 14 French hospital laboratories . METHODS: During the second quarter of 1996, 1997 and 1998, 180 consecutive non-duplicate isolates of Enterobacteriaceae were collected in each center . Sixteen beta-lactams and four quinolones were tested by the disk diffusion method . In addition, the double-disk synergy test was used to screen for the production of extended-spectrum beta-lactamase (ESBL) . RESULTS: Totals of 2507, 2312 and 2506 clinical isolates were obtained in each period, respectively . The distribution of Enterobacteriaceae species according to clinical specimens and wards was similar in each study period . No significant variation in the susceptibility rates to beta-lactams and fluoroquinolones was observed, except in Klebsiella pneumoniae and Enterobacter aerogenes . The prevalence of ESBL-producing isolates decreased from 18% to 9% in the former, while it increased from 32% to 54% in the latter . At the same time, the susceptibility to ofloxacin and pefloxacin increased for K . pneumoniae (P < 0.003) and cephalosporinase-producing species (P < 0.05), except Enterobacter spp . CONCLUSION: Over the 3-year study period beta-lactams and fluoroquinolones remained highly active against Enterobacteriaceae clinical isolates, with the exception of E . aerogenes, probably as a result of the dissemination of multiresistant clones in French hospitals. J Endod, 2002 Apr, 28(4), 304 - 10 Microbiota of periapical lesions refractory to endodontic therapy; Sunde PT et al.; The periapical microbiota of 36 teeth with refractory apical periodontitis was investigated . None of the teeth had responded to conventional endodontic or long-term (> 6 months), calcium-hydroxide treatment . Eight patients had received antibiotics systemically . After anaerobic culture, a total of 148 microbial strains were detected among 67 microbial species . One of the 36 lesions was culture-negative . Approximately half (51.0%) of the bacterial strains were anaerobic . Gram-positive species constituted 79.5% of the flora . Facultative organisms, such as Staphylococcus, Enterococcus, Enterobacter, Pseudomonas, Stenotrophomonas, Sphingomonas, Bacillus, or Candida species were recovered from 27 of the lesions (75%) . Sulfur granules were found in 9 lesions (25%) . In these granules Actinomyces israelii, A . viscosus, A . naeslundii, and A . meyeri were identified . Other bacterial species, both gram-positive and gram-negative, were detected in the granules as well . Two sulfur granules did not contain Actinomyces . Scanning electron microscopy demonstrated rod- and spirochete-like cells in the granules, and transmission electron microscopy revealed organisms with copious amounts of extracellular material . Outer membrane vesicles were also seen . Some of the granules were calcified . This study demonstrated a wide variety of microorganisms, particularly gram-positive ones, in the periapical lesions of teeth with refractory apical periodontitis. Zh Mikrobiol Epidemiol Immunobiol, 2002 Mar-Apr, (2), 73 - 5 {Microflora of the inflammatory erosive areas of the esophagus in esophagitis patients}; Chervinets VM; Seven patients with erosive esophagitis and reflux esophagitis were examined . In cases of inflammatory erosive phenomena staphylococci, Micrococcus luteus, Candida, bacteria of the genera Pseudomonas, Veilonella, Klebsiella and other bacteria of the family Enterobacteriaceae, as well as Helicobacter pylori were detected in different frequency . In most cases concentrations of microorganisms were 4.07-5.39 Ig CFU/g . Isolated microorganisms producing different pathogenicity enzymes--hemolysin (Streptococcus intermedius, S . sanguis, Staphylococcus saprophyticus, S . warneri, Bacteroides spp.), lecithinase (Staphylococcus xylosus), caseinase, RNAase and catalase--were detected. Zh Mikrobiol Epidemiol Immunobiol, 2002 Mar-Apr, (2), 119 - 20 {Etiology of purulent septic diseases and the antibiotic resistance of the isolated causative agents}; Mefod'ev VV et al.; Among the causative agents of purulent septic diseases in the surgical hospital, 25 microbial species were isolated; of these, the prevailing species were Staphylococcus aureus (19.86 +/- 1.07%), Escherichia coli (16.5 +/- 0.99%) and Pseudomonas aeruginosa (10.06 +/- 0.8%) . From environmental objects in the hospital 14 microbial species were isolated, among them bacteria of the genus Enterobacter (27 +/- 1.7%), E . coli (19.07 +/- 1.48%), S . aureus (14.7 +/- 1.31%), Klebsiella pneumoniae (13.73 +/- 1.31%), P . aeruginosa (7.33 +/- 0.98%) . During 3 years of observation the isolation rate of K . pneumoniae from different environmental objects was found to increase threefold to 24.7 +/- 2.7% . The results of the study of the microbial picture in surgical hospitals, as well as the antibiotic resistance of circulating causative agents, should be borne in mind while taking epidemic control measures. J Chemother, 1991 Jan, 3 Suppl 1, 54 - 6 Interaction with CEP-1 beta-lactamase of RU 51746-2, the active form of the new oral cephalosporin RU 51807; Dusi C et al.; RU 51746-2 (which is the water soluble Na-salt of RU 51763) proved stable toward the most common beta-lactamases tested, with the sole exception of CEP-1, a plasmid-determined enzyme which is homologous to chromosomal beta-lactamases of some enterobacterial species . We studied the interaction of RU 51746-2 with CEP-1 in an E . coli C600 derivative containing pNU104, a multicopy plasmid in which an up-promoter mutation has increased the level of beta-lactamase expression . We found that RU 51746-2 was indeed hydrolyzed with slow, but significant, rates, which might account for the high minimum inhibitory concentration (MIC) values (> 128 microg/ml) we found for this laboratory strain . However, when the same test was performed with an 100-fold dilution of the sonic extract, in order to obtain enzyme levels comparable to those usually found in clinical isolates, no appreciable hydrolysis could be measured . This suggests that the slow hydrolysis of RU 51746-2 has no clinical meaning, since the activity of this drug is virtually unaffected by the amount of beta-lactamases usually found in wild strains. J Chemother, 1991 Jan, 3 Suppl 1, 190 - 5 Bacterial adhesion to mucosal surfaces; Taylor RK; Bacteria utilize one or more of several mechanisms to adhere to mucosal surfaces . Members of the enterobacteriaceae and related bacteria typically elaborate adhesive filamentous appendages from their surfaces termed fimbriae (or pili) . Vibrio cholerae expresses at least three types of these structures that are likely involved in various stages of colonizing the intestine . So far one of these, termed TCP for toxin coregulated pilus, has been shown to be essential for colonization and virulence in humans and in an infant mouse model . The major pilin subunit, TcpA, is a member of the type 4 pilin class, common to a number of diverse pathogenic bacteria . To understand how these pili function at the molecular level, a transposon mutagenesis was undertaken which identified at least a dozen genes involved in TCP biogenesis, function, and regulation . Despite this complexity, passive immunization of infant mice using polyclonal and monoclonal antibodies directed against TCP suggests that the adhesive moiety lies within the major pilin subunit itself, rather than as a minor pilus adhesin protein as in the case of the PAP and type 1 fimbriae of Escherichia coli . Pilin or peptides corresponding to the epitopes recognized by the monoclonal antibodies represent possible immunogens for improved cholera vaccines. Am J Orthop, 2002 May, 31(5), 264 - 7 Superinfection of chronic multifocal osteomyelitis with Enterobacter cloacae; Pneumaticos SG et al.; A case is presented of a 6-year-old boy from Nigeria who developed chronic multifocal osteomyelitis with bilateral and symmetrical proximal humeral involvement . Superimposed changes of acute osteomyelitis were noted on the histopathological evaluation of the lesions . Cultures from both sides were positive for Enterobacter cloacae, which was presumably contracted during intravenous therapy for typhoid fever in Nigeria 1 month prior to his presentation. J Antimicrob Chemother, 2002 Jun, 49(6), 989 - 97 Beta-lactamases involved in resistance to broad-spectrum cephalosporins in Escherichia coli and Klebsiella spp . clinical isolates collected between 1994 and 1996, in Barcelona (Spain); Sabate M et al.; The aim of this study was to evaluate the incidence of decreased susceptibility to broad-spectrum cephalosporins in Enterobacteriaceae that lack inducible chromosomal bla genes, and to determine the enzymes responsible for resistance . From all clinically relevant Enterobacteriaceae strains isolated between 1994 and 1996, 88 of 7054 Escherichia coli, seven of 581 Klebsiella pneumoniae and 23 of 166 Klebsiella oxytoca strains were studied because of their decreased susceptibilities to broad-spectrum cephalosporins (as reflected in intermediate susceptibilities and/or positive synergy tests and/or irregular crenellated inhibition zones) . The most frequent mechanism implicated in decreased susceptibility to broad-spectrum cephalosporins displayed by E . coli and K . oxytoca was hyperproduction of chromosomal beta-lactamase, followed by plasmid-mediated SHV-1 hyperproduction in E . coli . In our hospital, the incidence of plasmid-mediated extended-spectrum beta-lactamases (ESBLs) between 1994 and 1996 was low . ESBLs were found in only 10 (0.14%) E . coli strains (six CTX-M-9, two TEM-12 and two SHV-2), in one (0.17%) K . pneumoniae strain (SHV-2) and in no K . oxytoca strains . The relatively wide variety of beta-lactamases that were detected among these common bacteria isolated from a single medical centre, including non-TEM- and non-SHV-derived ESBLs, appears epidemiologically remarkable. J Clin Microbiol, 2002 Jun, 40(6), 2057 - 61 Molecular subtyping methods for detection of Salmonella enterica serovar Oranienburg outbreaks; Kumao T et al.; This study involved 82 Salmonella enterica serovar Oranienburg isolates from patients with gastroenteritis and/or focal infections, healthy carriers, and cuttlefish chips which were epidemiologically linked to a major outbreak that had affected 1,505 people in Japan between 1998 and 1999 . We concurrently investigated four different molecular subtyping methods using human salmonellosis-associated Salmonella serovars and their applicability in detection of serovar Oranienburg in an outbreak . Pulsed-field gel electrophoresis (PFGE), enterobacterial repetitive intergenic sequence PCR (ERIC2-PCR), or 16S/23S rRNA ribotyping provided a high degree of interserovar discrimination for most of the serovars, with PFGE being the most discriminatory . For intraserovar typing of serovar Oranienburg, ERIC2-PCR was found to be the most sensitive . Native plasmid profiling, however, revealed nine different subgroups among epidemiologically and genetically related outbreak strains . Using these methods, a link was confirmed between food (cuttlefish chips) and patients in the serovar Oranienburg outbreak . This study underscores the limitations of chromosome-based and plasmid-based typing methods. Curr Microbiol, 2002 Jul, 45(1), 41 - 5 Transformed bacterial symbionts re-introduced to and detected in host gut; Peloquin JJ et al.; Strains of Enterobacter agglomerans and Klebsiella pneumoniae isolated from Rhagoletis completa Cresson were engineered to express transgenic fluorescent proteins (ECFP, DsRed) . These bacteria were introduced into flies by feeding the flies a sucrose solution in which the bacteria were suspended . The transgenic and heterologous marker protein was expressed and visible in the bacteria after they were ingested by WHF and while they were in the fly gut . We describe the plasmids used to transform these bacteria and demonstrate expression of heterologous proteins from the transforming plasmids and discuss the implications for future pest control strategies. Microb Ecol, 2001 Oct, 42(3), 286 - 294 Spatial and Temporal Variation of Enterobacter Genotypes in Sediments and the Underlying Hyporheic Zone of an Agricultural Stream; Halda-Alija L et al.; Population studies of enteric bacteria in an agriculturally impacted stream (Ledbetter Creek, Murray, Kentucky, USA) were conducted over a period of 2 years . Total number of bacteria, cultivated heterotrophic aerobic bacteria, and enteric bacteria showed significant differences between winter and summer . The cultivated numbers of heterotrophic aerobic bacteria and enteric bacteria were significantly more abundant in summer than in winter . The abundance of enteric bacteria was 12.9% in an upwelling zone and 9.8% in a downwelling zone in summer . Most of the enteric bacterial strains isolated on MacConkey agar were assigned to Enterobacter cloacae and E . agglomerans by API 20E and an analysis of the restriction patterns produced by amplified DNA coding for 16S rRNA (ARDRA) with the enzyme Hpa II . E . cloacae and E . agglomerans genotypes isolated from three hyporheic and gravel bar depth intervals (0-10 cm, 15-25 cm, and 30-40 cm) in summer and fall showed significant spatial variation and were heterogeneously distributed along the stream . Temperature, inorganic nutrients, and occurrence of anoxic zones affected the distribution of enteric bacteria . These techniques can be used as a model to monitor shifts among different species in the stream ecosystem. FEMS Microbiol Lett, 2002 Apr 23, 210(1), 87 - 92 Chromosomal ampC genes in Enterobacter species other than Enterobacter cloacae, and ancestral association of the ACT-1 plasmid-encoded cephalosporinase to Enterobacter asburiae; Rottman M et al.; The amplification and sequence of ampC genes in Enterobacter asburiae, Enterobacter cancerogenus, Enterobacter dissolvens, Enterobacter hormaechei and Enterobacter intermedius bring the number of known cephalosporinase sequences from the genus Enterobacter to seven . Expression in Escherichia coli of the ampC genes from E . asburiae, E . hormaechei and E . intermedius established the functional nature of these genes . ampC from E . asburiae shows 96.5% identity to bla(ACT-1) encoding a plasmid-borne cephalosporinase previously believed to derive from Enterobacter cloacae . The reassignment of ACT-1 ancestry to E . asburiae is confirmed by the 95.5% identity between ampR upstream of bla(ACT-1) and ampR from E . asburiae. Antimicrob Agents Chemother, 2002 Jun, 46(6), 1966 - 70 Mutational replacement of Leu-293 in the class C Enterobacter cloacae P99 beta-lactamase confers increased MIC of cefepime; Vakulenko SB et al.; The class C beta-lactamase from Enterobacter cloacae P99 confers resistance to a wide range of broad-spectrum beta-lactams but not to the newer cephalosporin cefepime . Using PCR mutagenesis of the E . cloacae P99 ampC gene, we obtained a Leu-293-Pro mutant of the P99 beta-lactamase conferring a higher MIC of cefepime (MIC, 8 microg/ml, compared with 0.5 microg/ml conferred by the wild-type enzyme) . In addition, the mutant enzyme produced higher resistance to ceftazidime but not to the other beta-lactams tested . Mutants with 15 other replacements of Leu-293 were prepared by site-directed random mutagenesis . None of these mutant enzymes conferred MICs of cefepime higher than that conferred by Leu-293-Pro . We determined the kinetic parameters of the purified E . cloacae P99 beta-lactamase and the Leu-293-Pro mutant enzyme . The catalytic efficiencies (k(cat)/K(m)) of the Leu-293-Pro mutant beta-lactamase for cefepime and ceftazidime were increased relative to the respective catalytic efficiencies of the wild-type P99 beta-lactamase . These differences likely contribute to the higher MICs of cefepime and ceftazidime conferred by this mutant beta-lactamase. Antimicrob Agents Chemother, 2002 Jun, 46(6), 1665 - 70 Pharmacodynamics of the new fluoroquinolone gatifloxacin in murine thigh and lung infection models; Andes D et al.; Gatifloxacin is a new 8-methoxy fluoroquinolone with enhanced activity against gram-positive cocci . We used the neutropenic murine thigh infection model to characterize the time course of antimicrobial activity of gatifloxacin and determine which pharmacokinetic (PK)-pharmacodynamic (PD) parameter best correlated with efficacy . The thighs of mice were infected with 10(6.5) to 10(7.4) CFU of strains of Staphylococcus aureus, Streptococcus pneumoniae, or Escherichia coli, and the mice were then treated for 24 h with 0.29 to 600 mg of gatifloxacin per kg of body weight per day, with the dose fractionated for dosing every 3, 6, 12, and 24 h . Levels in serum were measured by microbiologic assay . In vivo postantibiotic effects (PAEs) were calculated from serial values of the log(10) numbers of CFU per thigh 2 to 4 h after the administration of doses of 8 and 32 mg/kg . Nonlinear regression analysis was used to determine which PK-PD parameter best correlated with the numbers of CFU per thigh at 24 h . Pharmacokinetic studies revealed peak/dose values of 0.23 to 0.32, area under the concentration-time curve (AUC)/dose values of 0.47 to 0.62, and half-lives of 0.6 to 1.1 h . Gatifloxacin produced in vivo PAEs of 0.2 to 3.1 h for S . pneumoniae and 0.4 to 2.3 h for S . aureus . The 24-h AUC/MIC was the PK-PD parameter that best correlated with efficacy (R(2) = 90 to 94% for the three organisms, whereas R(2) = 70 to 81% for peak level/MIC and R(2) = 48 to 73% for the time that the concentration in serum was greater than the MIC) . There was some reduced activity when dosing every 24 h was used due to the short half-life of gatifloxacin in mice . In subsequent studies we used the neutropenic and nonneutropenic murine thigh and lung infection models to determine if the magnitude of the AUC/MIC needed for the efficacy of gatifloxacin varied among pathogens (including resistant strains) and infection sites . The mice were infected with 10(6.5) to 10(7.4) CFU of four isolates of S . aureus (one methicillin resistant) per thigh, nine isolates of S . pneumoniae (two penicillin intermediate, four penicillin resistant, and two ciprofloxacin resistant) per thigh, four isolates of the family Enterobacteriaceae per thigh, a single isolate of Pseudomonas aeruginosa per thigh, and 10(8.3) CFU of Klebsiella pneumoniae per lung . The mice were then treated for 24 h with 0.29 to 600 mg of gatifloxacin per kg every 6 or 12 h . A sigmoid dose-response model was used to estimate the dose (in milligrams per kilogram per 24 h) required to achieve a net bacteriostatic effect over 24 h . MICs ranged from 0.015 to 8 microg/ml . The 24-h AUC/MICs for each static dose (1.7 to 592) varied from 16 to 72 . Mean +/- standard deviation 24-h AUC/MICs for isolates of the family Enterobacteriaceae, S . pneumoniae, and S . aureus were 41 +/- 21, 52 +/- 20, and 36 +/- 9, respectively . Methicillin, penicillin, or ciprofloxacin resistance did not alter the magnitude of the AUC/MIC required for efficacy . The 24-h AUC/MICs required to achieve bacteriostatic effects against K . pneumoniae were quite similar in the thigh and lung (70 versus 56 in neutropenic mice and 32 versus 43 in nonneutropenic mice, respectively) . The magnitude of the 24-h AUC/MIC of gatifloxacin required for efficacy against multiple pathogens varied only fourfold and was not significantly altered by drug resistance or site of infection. Virus Genes, 2002 Mar, 24(2), 163 - 71 Evolution of lambdoid replication modules; Wrobel B et al.; Comparison of the putative iteron-binding proteins of lambdoid phages allows us to propose that in the case of lambdoid replication modules, the units on which natural selection acts do not coincide with the open reading frames . Rather, the first replication gene is split into two segments, and its 3' part (corresponding to the C-terminal domain of the iteron-binding protein) forms one unit with the second gene . We also propose from the phylogenetic analysis of phage-encoded homologs of E . coli DnaB and DnaC, that the recombination with the host sequences is not frequent . Accessory ATP-ases for helicase loading (E . coli DnaC homologs) may not be universal replication proteins . Our analysis may suggest that the bacterial helicase loaders might be of phage origin . The comparison of DnaC homologs of enterobacteria and enterobacterial phages supports the experimental data on residues important in interaction with DnaB . We propose that construction of plasmids carrying the replication origins of lambdoid prophages could be useful not only in further research on DNA replication but also on the role of these prophages in shuttling genes for bacterial virulence . The phage replication sequences could be also useful for identification of clinical enterobacterial isolates. Nature, 2002 May 2, 417(6884), 91 - 4 Neutrophil elastase targets virulence factors of enterobacteria; Weinrauch Y et al.; Shigellae cause bacillary dysentery, a bloody form of diarrhoea that affects almost 200 million people and causes nearly 2 million deaths per year . Shigella invades the colonic mucosa, where it initiates an acute inflammation, rich in neutrophils, that initially contributes to tissue damage and eventually resolves the infection . Neutrophils are phagocytic cells that kill microorganisms but it is unclear how neutrophils control pathogenic bacteria expressing virulence factors that manipulate host cells . In contrast to other cells, neutrophils prevent the escape of Shigella from phagocytic vacuoles in which the bacteria are killed . Here we identify human neutrophil elastase (NE) as a key host defence protein: NE degrades Shigella virulence factors at a 1,000-fold lower concentration than that needed to degrade other bacterial proteins . In neutrophils in which NE is inactivated pharmacologically or genetically, Shigella escapes from phagosomes, increasing bacterial survival . NE also preferentially cleaves virulence factors of Salmonella and Yersinia . These findings establish NE as the first neutrophil factor that targets bacterial virulence proteins. Hunan Yi Ke Da Xue Xue Bao, 1999, 24(3), 299 - 300 {A modified MacConkey medium}; Wang J et al.; We used MacConkey Agar(MCA) medium powder as a basis and added 6-benzoy-2-naphthyl phosphate disodium salt to modify MCA medium . This method was identified by examining several kinds of enteric pathogenic bacteria . Thus it allows us to observe lactose fermentative and phosphatase reaction simultaneously and to recognize Enterobacteriaceae from non-fermentable bacteria on primary culture plates according to the color of colony . This improved method simplifies the recognition procedure and shortens the time so that it is helpful to correctly choosing identification kit. Ann N Y Acad Sci, 2002 Apr, 958, 408 - 11 IgM, IgG, and IgA response to enterobacteria in patients with ankylosing spondylitis in southern India; Madhavan R et al.; IgM, IgA, and IgG response to three different antigenic preparations-lipopolysaccharide (LPS), culture supernatant proteins, and outer membrane protein (OMP) of Klebsiella pneumoniae, Escherichia coli and Salmonella typhi-were measured in the sera of 20 patients with primary ankylosing spondylitis (AS), 10 with enterogenic reactive arthritis (ReA) (disease controls), and 15 voluntary blood donors (healthy controls) by ELISA using biotinylated anti-human immunoglobulins M, G, and A . Serum immunoglobulin levels were measured by immunoturbidimetric assay in 20 AS patients, 20 patients with enterogenic reactive arthritis (ReA), 20 with ulcerative colitis (UC), and 20 voluntary blood donors . Student's t-test was applied for comparison . Compared to healthy controls, AS patients showed significantly elevated IgG response against culture supernatant proteins of all the three organisms (P <0.05), LPS of E . coli (P < 0.05) and Klebsiella (P < 0.005), as well as OMP only of Klebsiella pneumoniae . This was reflected as significantly elevated IgG level in AS compared to controls (P < 0.05 vs . ReA and 0.005 vs . UC and healthy controls) . This suggests the involvement of outer membrane proteins of Klebsiella pneumoniae in the pathogenic mechanism of ankylosing spondylitis. N Engl J Med, 2002 May 16, 346(20), 1529 - 37 Serratia marcescens bacteremia traced to an infused narcotic; Ostrowsky BE et al.; BACKGROUND: From June 30, 1998, through March 21, 1999, several patients in the surgical intensive care unit of a hospital acquired Serratia marcescens bacteremia . We investigated this outbreak . METHODS: A case was defined as the occurrence of S . marcescens bacteremia in any patient in the surgical intensive care unit during the period of the epidemic . To identify risk factors, we compared patients with S . marcescens bacteremia with randomly selected controls . Isolates from patients and from medications were evaluated by pulsed-field gel electrophoresis . The hair of one employee was tested for fentanyl . RESULTS: Twenty-six patients with S . marcescens bacteremia were identified; eight (31 percent) had polymicrobial bacteremia, and seven of these had Enterobacter cloacae and S . marcescens in the same culture . According to univariate analysis, patients with S . marcescens bacteremia stayed in the surgical intensive care unit longer than controls (13.5 vs . 4.0 days, P<0.001), were more likely to have received fentanyl in the surgical intensive care unit (odds ratio, 31; P<0.001), and were more likely to have been exposed to two particular respiratory therapists (odds ratios, 13.1 and 5.1; P<0.001 for both comparisons) . In a multivariate analysis, receipt of fentanyl and exposure to the two respiratory therapists (adjusted odds ratio for one therapist, 6.7; P=0.002; adjusted odds ratio for the other therapist, 9.5; P=0.02) remained significant . One respiratory therapist had been reported for tampering with fentanyl; his hair sample tested positive for fentanyl . Cultures of fentanyl infusions from two case patients yielded S . marcescens and E . cloacae . The isolates from the case patients and from the fentanyl infusions had similar patterns on pulsed-field gel electrophoresis . After removal of the implicated respiratory therapist, no further cases occurred . CONCLUSIONS: An outbreak of S . marcescens and E . cloacae bacteremia in a surgical intensive care unit was traced to extrinsic contamination of the parenteral narcotic fentanyl by a health care worker . Our findings underscore the risk of complications in patients that is associated with illicit narcotic use by health care workers. Rhinology, 2002 Mar, 40(1), 18 - 23 Bacteriology of chronic sinusitis: the bulla ethmoidalis content; Rombaux P et al.; OBJECTIVE: To study the microbiology of the bulla ethmoidalis of patients who suffered from chronic sinusitis . DESIGN: A prospective study performed at the Saint-Luc universitary clinic (University of Louvain) from June 1999 to December 2000 . MATERIAL AND METHODS: Seventy seven patients underwent functional endoscopic endonasal surgery during this period for chronic sinusitis refractory to standard medical treatment . All the patients presented symptoms for more than 3 months . After Povidone-Iodine disinfection, samples were taken into the bulla ethmoidalis after its opening with an endoscopic endonasal control . Samples were transferred to the microbiology laboratory using a thioglycolate medium for aerobic and anaerobic cultures . RESULTS: One hundred forty eight samples were studied . Culture rate was 73.6% . Thirty nine samples remained sterile . In the 109 culture positive specimens, 135 bacterial isolates were recovered . The main results are: Staphylococcus coagulase negative: 31, Staphylococcus aureus: 22, Streptococcus sp: 20, other Gram positive Cocci: 5, Haemophilus influenzae: 4, non-fermentative Gram negative bacilli: 6, Enterobacteriaceae: 45, anaerobic bacteria: 2 . CONCLUSION: Enterobacteriaceae or enteric gram negative bacilli were frequently encountered in the bulla ethmoidalis of patients suffering from chronic sinusitis . This report suggests that endoscopically guided culture obtained from the ethmoid sinus may accurate our understanding of the microbiology of chronic ethmoidal sinusitis and underline the importance of Enterobacteriaceae in this disease. Dermatology, 2002, 204 Suppl 1, 79 - 85 Efficacy of povidone-iodine vaginal suppositories in the treatment of bacterial vaginosis; Wewalka G et al.; A prospective, randomized clinical trial was performed to study the efficacy of povidone iodine (Betadine) suppositories for the treatment of bacterial vaginosis (BV) in comparison to capsules containing lactobacilli (Doderlein Med) . Seventy patients with BV included in the study were randomly assigned to be either treated with povidone iodine suppositories or lactobacilli . Patients were treated once a day for 5 days . Initial examinations took place on the first day of the study with follow-up examinations on days 8 and 15 . The examinations included clinical parameters, patient evaluation, secretion screens, and quantitative and qualitative microbiological tests of vaginal flora . Both treatment groups showed improvement of clinical parameters, condition of secretions and subjective state of health . At day 15 there was a trend towards a better efficacy of the treatment with povidone iodine but this was not significant . However, patients with acute BV treated with povidone iodine had significantly better scores after 15 days . Both treatments were well tolerated . The microbiological examinations showed an increase of the mean number of lactobacilli in the vagina on day 8 after initiation of treatment with lactobacilli, but a decrease on day 15 . Contrary to that the lactobacilli counts from patients treated with povidone iodine suppositories decreased after the first week but increased in the second one . Potentially pathogenic germs, e.g . Gardnerella, Bacteroides and Enterobacteria were reduced in a higher extent and with a longer lasting effect after treatment with povidone iodine suppositories than with capsules containing lactobacilli . The results of this study show that native lactobacilli rapidly re-colonize after the antiseptic treatment with povidone iodine . Therefore, there is no need to use lactobacilli in addition . J Hosp Infect, 2002 May, 51(1), 21 - 6 Management of an outbreak of Enterobacter cloacae in a neonatal unit using simple preventive measures; v Dijk Y et al.; Enterobacter cloacae is becoming an increasingly important nosocomial pathogen . Outbreaks of E . cloacae in intensive care units and burns units have been described frequently . In December 1999, a neonate with line sepsis was transferred from a university hospital to the neonatal unit of the Diakonessen Hospital . Blood culture yielded E . cloacae . An outbreak of E . cloacae was occurring in the university hospital at that time . In February 2000, a second neonate in our hospital developed line sepsis caused by E . cloacae . Direct measures taken included cohorting of infected children, disinfection of incubators, thermometers and wards, and screening patients . Of nine neonates, seven were colonized with E . cloacae . Despite these measures, the outbreak continued . Forty-one patients were screened; 15 were colonized . Environmental searches yielded E . cloacae in a sink and on two thermometers . Sixteen isolates were typed by arbitrarily primed PCR using four primers . All the patient isolates and the two isolates from thermometers were identical . The strain isolated from the sink was unrelated . Amplified fragment length polymorphism typing showed that the outbreak clone was identical to that in the university hospital . After the introduction of disposable thermometer covers, E . cloacae colonization slowly decreased . J Invertebr Pathol, 2001 Nov, 78(4), 232 - 43 Pathobiology of amber disease, caused by Serratia Spp., in the New Zealand grass grub, Costelytra zealandica; Jackson TA et al.; Amber disease in the New Zealand grass grub (Costelytra zealandica) is caused by some strains of Serratia entomophila or Serratia proteamaculans (Enterobacteriaceae) . When treated with pathogenic isolates, larvae ceased feeding within 48 h, developed an amber coloration after 72 h, and entered a long chronic phase without feeding . An acute dose of 2-4 x 10(4) pathogenic bacteria was sufficient to produce disease in 50% of treated larvae . Time to death was directly related to temperature . At 15 degrees C, infected larvae remained in a chronic, nonfeeding state for more than 4 months prior to death . Nonpathogenic isolates, lacking the disease-causing plasmid (pADAP), had no effect on either feeding or disease . Twenty-four hours after ingestion, bacteria were found predominantly in the hindgut and growth occurred primarily within the fermentation chamber and in the head section of the larvae . Nonpathogenic strains did not multiply in treated larvae . Treatment of diseased larvae with antibiotic eliminated Serratia cells from the insects but did not result in restoration of feeding or the dark gut characteristic of the healthy larva. Int J Med Microbiol, 2002 Mar, 291(8), 649 - 54 Cloning and sequencing of Enterobacter aerogenes OmpC-type osmoporin linked to carbapenem resistance; Sturenburg E et al.; Using outbreak-related strains of Enterobacter aerogenes, we cloned and sequenced ompK39, the structural gene coding for outer membrane protein OmpK39 . Its lack of expression was closely associated with a phenotype exhibiting low-level carbapenem resistance . Detailed alignment of the predicted amino acid sequence revealed that OmpK39 is a member of the OmpC subclass of enterobacterial porins, with the highest degree of homology to Klebsiella pneumoniae OmpK36 . Based on a computerized alignment including Escherichia coli PhoE and OmpF, the 3D structures of which are known from X-ray studies, OmpK39 can be assumed to form the typical beta-barrel structure which is common to all enterobacterial porins . Since no inhibitory DNA sequences could be detected in ompk39 in the resistant strains, porin deficiency leading to carbapenem resistance seems to involve alterations in key regulatory genes and/or the promotor sequence rather than a direct mutation in the structural gene. Int J Antimicrob Agents, 2002 May, 19(5), 431 - 4 Susceptibility patterns of Gram-negative bacteria from a Polish intensive care unit, 1997-2000; Patzer J et al.; The susceptibility of Gram-negative bacterial strains (n=400) isolated from clinical specimens of children hospitalized in a Polish intensive care unit (ICU) between 1997 and 2000 was tested . Meropenem, imipenem and ciprofloxacin were most active (>90% susceptibility) against the tested isolates, with no observed reduction in activity over 4 years . Extended-spectrum beta-lactamases and AmpC beta-lactamase producers among Enterobacteriaceae isolates from this ICU continued to be a serious therapeutic problem, although the carbapenems were highly active against these resistance phenotypes . Resistance to aminoglycosides (gentamicin, tobramycin) and ceftazidime was a characteristic of >40% of tested isolates. Int J Antimicrob Agents, 2002 May, 19(5), 397 - 404 Growth of cell-wall-deficient variants of Enterobacter cloacae facilitates beta-lactamase derepressed mutants; Huber TW; The degree to which cell-wall-deficient bacteria (CWDB) are involved in the generation of beta-lactamase derepressed mutants (DM) was measured using Enterobacter cloacae 3624 . The frequency of DM in non-permissive isotonic ticarcillin medium was compared with their frequency in hypertonic ticarcillin medium that supports CWDB growth . DM were resistant to extended spectrum penicillins and cephalosporins and had a basal beta-lactamase activity of >300 units/mg protein . Anaerobic growth of CWDB increased the relative risk of DM 2 x 10(6)-fold . Aerobic incubation produced fewer CWDB colonies but the risk of DM was still increased 400-fold over non-permissive controls . These results define a new role for CWDB as intermediaries in the emergence of resistance. FEMS Microbiol Lett, 2002 Apr 9, 209(2), 161 - 8 Diversity of CTX-M beta-lactamases and their promoter regions from Enterobacteriaceae isolated in three Parisian hospitals; Saladin M et al.; Nine clinical isolates of Enterobacteriaceae (six Escherichia coli and three Proteus mirabilis) isolated in three Parisian hospitals between 1989 and 2000 showed a particular extended-spectrum cephalosporin-resistance profile characterized by resistance to cefotaxime and aztreonam but not to ceftazidime . CTX-M-1, CTX-M-2, CTX-M-9, CTX-M-14 and two novel plasmid-mediated CTX-M beta-lactamases (CTX-M-20, and CTX-M-21) were identified by polymerase chain reaction and isoelectric focusing (pI>8) and were associated in eight cases with TEM-1 (pI=5.4) or TEM-2 (pI=5.6) beta-lactamases . We used internal ISEcp1 and IS26 forward primers and the CTX-M consensus reverse primer to characterize the CTX-M beta-lactamase promoter regions and showed their high degree of structure diversity . We found upstream of some bla(CTX-M) genes, a 266-bp sequence 100% identical to the sequence upstream of the Kluyvera ascorbata beta-lactamase gene, suggesting that this chromosomal enzyme is the progenitor of the CTX-M-2/5 cluster. Diagn Microbiol Infect Dis, 2002 Apr, 42(4), 257 - 61 High prevalence of nalidixic acid resistant, ciprofloxacin susceptible phenotype among clinical isolates of Escherichia coli and other Enterobacteriaceae; Ruiz J et al.; Therapeutic failure of infections during their treatment with quinolones has been often described . This may be due to the development of resistance during treatment of an infecting strain which already had diminished susceptibility to quinolones, even though the initial MIC did not exceed the breakpoint . In this study the prevalence of the nalidixic acid resistant, ciprofloxacin susceptible phenotype among Enterobacteriaceae was analyzed . The results showed that 113 out of 151 (74.83%) strains of the Enterobacteriaceae with diminished susceptibility to ciprofloxacin (MICs from 0.06 to 1 microg/ml) were resistant to nalidixic acid (MICs > 32 microg/ml) . The Escherichia coli strains presenting this phenotype already have a mutation in the amino acid codon Ser-83 of the gyrA gene, so that the possibility of developing a second mechanism of resistance during treatment is very high. Enferm Infecc Microbiol Clin, 2002 May, 20(5), 225 - 34 {Interpretive reading of the antibiogram of enterobacteria}; Navarro Risueno F et al.; Many of the resistance mechanisms of enterobacteria to antimicrobial agents are well understood; nevertheless several aspects remain unsolved, particularly with regard to prediction of clinical response . The resistance pattern observed in the antibiogram of a specific organism should be the sum of the natural resistance pattern, characteristic of the species, plus the acquired resistance . In enterobacteria the principal mechanism of resistance to beta lactams and aminoglycosides is enzyme production, Each enzyme recognizes one or more specific beta lactam or aminoglycoside, as a substrate . This translates as a specific resistance phenotype that allows one to infer the enzyme(s) implicated . Enzyme resistance is not, however, the only mechanism of resistance to these agents; often the pattern observed is multifactorial . Resistance to quinolones is due to point and sequence mutations which may be selected by initially active fluoroquinolones and cause a stepwise increase of resistance. J Environ Health, 2002 May, 64(9), 9 - 16, 32; quiz 35-6 Salmonella and other Enterobacteriaceae in dairy-cow feed ingredients: antimicrobial resistance in western Oregon; Kidd RS et al.; Several studies have suggested an association between the use of antimicrobial agents in animal feeds and an increased risk that humans will contract resistant strains of bacteria such as Salmonella species, Escherichia coli, and other enteric isolates . The authors of this study evaluated whether animal feeds might serve as sources of antimicrobial-resistant bacteria, especially bacteria that are pathogenic to humans . From July through August 1998, samples of feed ingredients were collected from a total of 50 feed piles located on 12 dairy farms in western Oregon . From a subset of 10 piles, repeated samples were collected over time until each pile was depleted . Analysis of the samples indicated that 42.0 percent of all 50 piles and 60.0 percent of the piles from which there was repeated sampling were presumptive positive for Salmonella . Sixty-two percent of 50 Enterobacteriaceae isolates showed ampicillin resistance, and 10.0 percent displayed tetracycline resistance . Other bacteria displayed varying degrees of resistance to ampicillin, streptomycin, tetracycline, or a combination of these antimicrobials . The extent of antimicrobial-resistant Enterobacteriaceae in feed ingredients observed in this study raises significant concerns about the potential for human health risks from food-producing animals such as dairy cows. MMWR Morb Mortal Wkly Rep, 2002 Apr 12, 51(14), 297 - 300 Enterobacter sakazakii infections associated with the use of powdered infant formula--Tennessee, 2001; Active efflux et al.; Centre for Adaptation Genetics and Drug Resistance and the Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111, USA . stuart.levy@tufts.edu Energy-driven drug efflux systems are increasingly recognized as mechanisms of antibiotic resistance . Chromosomally located or acquired by bacteria, they can either be activated by environmental signals or by a mutation in a regulatory gene . Two major categories exist: those systems energized by proton motive force and those dependent on ATP . The pumps may have limited or broad substrates, the so-called multiple drug resistance pumps, which themselves form a number of related families . The multiple antibiotic resistance (mar) locus and mar regulon in Escherichia coli and other members of the Enterobacteriaceae is a paradigm for a generalized response locus leading to increased expression of efflux pumps . One such pump, the AcrAB pump extrudes biocides such as triclosan, chlorhexidine and quaternary ammonium compounds as well as multiple antibiotics . In Pseudomonas aeruginosa, a number of multidrug efflux pumps export a broad range of substrates . Since bacteria expressing these pumps thwart the efficacy of both kinds of therapeutic agents which control infectious diseases -- biocides which prevent transmission of infectious disease agents and antibiotics which treat and cure infectious diseases -- they are of particular concern . The prudent use of antibiotics and biocides will guard against the selection and propagation of drug-resistant mutants and preserve the efficacy of these valuable anti-infective agents. Int J Food Microbiol, 2002 May 5, 75(1-2), 119 - 26 Microbiological analysis of seed sprouts in Norway; Robertson LJ et al.; As part of larger survey of microbial contamination of fruits and vegetables in Norway, four different sprouted seed products were analysed for bacterial and parasitic contaminants (n = 300 for bacterial analyses and n = from 17 to 171 for parasite analyses, depending on parasite) . Escherichia coli O157, Salmonella, Listeria monocytogenes, Cyclospora oocysts, Ascaris eggs and other helminth parasites were not detected in any of the sprout samples . Thermotolerant coliform bacteria (TCB) were isolated from approximately 25% of the sprout samples, with the highest percentage of TCB positive samples in alfalfa sprouts . Most TCB were Enterobacter spp . and Klebsiella . E . coli was isolated from 8 of 62 TCB positive mung bean sprout samples . Cryptosporidium oocysts were detected in 8% of the sprout samples and Giardia cysts were detected in 2% of the samples . All the Cryptosporidium positive samples, and most of the Giardia positive samples, were mung bean sprouts . Parasite concentrations in positive samples were low (between 1 and 3 oocysts/cysts per 50 g sprouts) . Sprout irrigation water was also analysed for microbial contaminants . E . coli O157 and L . monocytogenes were not detected . TCB were isolated from approximately 40% of the water samples . Salmonella reading was isolated from three samples of spent irrigation water on 3 consecutive days . Cryptosporidium and Giardia were also isolated from spent irrigation water . Additionally, eight samples of unsprouted mung bean seed were analysed for Cryptosporidium oocysts and Giardia cysts . One or both of these parasites were detected in six of the unsprouted seed samples at concentrations of between 1 and 5 oocysts/cysts per 100 g unsprouted seed . Whilst our results support spent irrigation water as the most suitable matrix for testing for bacteria, unsprouted seed is considered the more useful matrix for analysing for parasite contaminants. Anesteziol Reanimatol, 2002 Jan-Feb, (1), 40 - 2 {Character and risk factors of bacterial colonization of newborns in intensive care units}; Mironov PI et al.; A total of 116 newborns treated in intensive care wards were examined . The incidence, type, and risk factors of bacterial colonization were evaluated . Only 7.8% patients were colonized with gram-negative polyresistant microflora by admission, while after 3 days of intensive care as many as 48.2% newborns were colonized . The major microorganisms were Pseudomonas aeruginosa, Klebsiella spp., and Enterobacter spp . A relationship was detected between the bacterial spectrum of the skin and mucosa and the agents of infectious inflammatory complications in the newborns . The most significant factors of risk of colonization in the newborns with antibiotic-resistant gram-negative bacteria in intensive care wards were forced ventilation of the lungs and initially severe clinical status (CRIB score 5 and higher). Dtsch Tierarztl Wochenschr, 2002 Apr, 109(4), 154 - 7 {Specificity of inhibition between Salmonella strains}; Martin G et al.; A phenomenon of inhibition among Salmonella organisms colonizing the gut of young animals can be demonstrated which is important in connection with immunization using live Salmonella vaccines . For practical utilization of this inhibition phenomenon, research into its specificity is very important . In vitro broth culture and animal experiments have shown inhibition to be genus-specific, i.e . there is no inhibition between strains of different genera of Enterobacteriaceae in most cases . The strongest inhibition occurs between isogenic (identical genotype) strains (strain-specific inhibition) . This applies to Salmonella and other genera of Enterobacteriaceae . There was a close correlation between the degree of the inhibitory effect and the genetic relatedness of the strains, as has been demonstrated in Salmonella strains of different epidemiological classification . The inhibitory capacity was most pronounced among closely related strains . Clonal strains of Salmonella Enteritidis phage type 4 fully inhibited caecal colonization by Salmonella Enteritidis phage type 4 strains in 65% of the combinations tested in chicks . In broth culture experiments, the share of combinations with complete inhibition was above 90% . A significant inhibitory competence of a selected strongly inhibitory Salmonella Enteritidis phage type 4 strain against the majority of Salmonella Enteritidis field isolates can be expected . However, Salmonella Enteritidis strains of all phase types showed almost no inhibitory competence against other Salmonella serotypes. Emerg Infect Dis, 2002 May, 8(5), 473 - 8 Trends in fluoroquinolone (ciprofloxacin) resistance in enterobacteriaceae from bacteremias, England and Wales, 1990-1999; Livermore DM et al.; The Public Health Laboratory Service receives antibiotic susceptibility data for bacteria from bloodstream infections from most hospitals in England and Wales . These data were used to ascertain resistance trends to ciprofloxacin from 1990 through 1999 for the most prevalent gram-negative agents: Escherichia coli, Klebsiella spp., Enterobacter spp., and Proteus mirabilis . Significant increases in resistance were observed for all four species groups . For E . coli, ciprofloxacin resistance rose from 0.8% in 1990 to 3.7% in 1999 and became widely scattered among reporting hospitals . The prevalence of resistance in Klebsiella spp . rose from 3.5% in 1990, to 9.5% in 1996 and 7.1% in 1999, while that in Enterobacter spp . rose from 2.1% in 1990 to 10.5% in 1996 and 10.9% in 1999 . For both Klebsiella and Enterobacter spp., most resistance was localized in a few centers . Resistance was infrequent and scattered in P . mirabilis, but reached a prevalence of 3.3% in 1999. Acta Biochim Pol, 2000, 47(4), 941 - 9 Two-electron reduction of nitroaromatic compounds by Enterobacter cloacae NAD(P)H nitroreductase: description of quantitative structure-activity relationships; Nivinskas H et al.; Enterobacter cloacae NAD(P)H:nitroreductase catalyzes the reduction of a series of nitroaromatic compounds with steady-state bimolecular rate constants (kcat/Km) ranging from 10(4) M(-1) s(-1) to 10(7) M(-1) s(-1), and oxidizing 2 moles NADH per mole mononitrocompound . Oxidation of excess NADH by polynitrobenzenes including explosives 2,4,6-trinitrotoluene (TNT) and 2,4,6-trinitrophenyl-N-methylnitramine (tetryl), has been observed as a slower secondary process, accompanied by O2 consumption . This type of 'redox cycling' was not related to reactions of nitroaromatic anion-radicals, but was caused by the autoxidation of relatively stable reaction products . The logs kcat/Km of all the compounds examined exhibited parabolic dependence on their enthalpies of single-electron- or two-electron (hydride) reduction, obtained by quantum mechanical calculations . This type of quantitative structure-activity relationships shows that the reactivity of nitroaromatics towards E . cloacae nitroreductase depends mainly on their hydride accepting properties, but not on their particular structure, and does not exclude the possibility of multistep hydride transfer. J AOAC Int, 2002 Mar-Apr, 85(2), 388 - 94 Validation of the Hygicult E dipslides method in surface hygiene control: a Nordic collaborative study; Salo S et al.; A collaborative study with Enterobacteriaceae was conducted to validate Hygicult E dipslides by comparison with violet red bile glucose agar (VRBGA) contact plates and swabbing, using stainless steel surfaces artificially contaminated with microbes at various levels . Twelve laboratories participated in the validation procedure . The total number of collaborative samples was 108 . The microbial level in each sample was assessed in triplicate by using the 3 above-mentioned methods . No Enterobacteriaceae were used at the low inoculation level . At the middle inoculation level, the percentages detached from the test surfaces were 16.6 with the Hygicult E method, 15.3 with the contact plate method, and 14.6 with swabbing; at the high innoculation level, the percentages were 14.5, 15.8, and 9.8, respectively . The percentage of acceptable results after the removal of outliers was 97.2 . Repeatability relative standard deviations ranged from 33.4 to 44.9%; reproducibility relative standard deviations ranged from 45.2 to 77.1% . The Hygicult E dipslide, VRBGA contact plate, and swabbing methods gave similar results at all 3 microbial levels tested: <1.0 colony-forming units (CFU)/cm2 at the low level, 1.2-1.3 CFU/cm2 at the middle level (theoretical yield 8.0 CFU/cm2), and 1.2-2.0 CFU/cm2 at the high level (theoretical yield 12.5 CFU/cm2). Pediatrics, 2002 May, 109(5), 846 - 51 Jaundice as an early diagnostic sign of urinary tract infection in infancy; Garcia FJ et al.; OBJECTIVE: Urinary tract infections (UTIs) are a common clinical problem in febrile infants younger than 8 weeks old, with a prevalence between 5% and 11% . Previous studies have noted that jaundice may be one of the first signs of a bacterial infection in infants . Our goal was to determine the incidence of UTIs in asymptomatic, jaundiced infants younger than 8 weeks old and to determine which historical and laboratory parameters are associated with UTIs . METHODS: We prospectively evaluated asymptomatic, jaundiced infants younger than 8 weeks old for evidence of a UTI (defined as >10 000 colony-forming units per milliliter of a single pathogen, obtained by bladder catheterization) . A serum fractionated bilirubin level was obtained on all study patients . Detailed questionnaires were completed, which included demographic information, prenatal, intrapartum, and postnatal events . RESULTS: Twelve (7.5%) of 160 infants had a UTI (95% confidence interval: 3.9%-12.7%) . Isolated organisms included Escherichia coli, Enterobacter cloacae, Enterococcus, Klebsiella pneumoniae, group B Streptococcus, Streptococcus viridans, and Staphylococcus aureus . Patients with the onset of jaundice after 8 days of age had a higher incidence of UTI, 6 (50%) of 12 infants in the positive culture group, versus 15 (10%) of 148 infants in the negative culture group . Abnormal urinalysis and microscopy results were noted in 5 (42%) of 12 infants with a UTI, and 6 (55%) of 11 infants had abnormal renal ultrasound results . CONCLUSION: A UTI was found in 7.5% of asymptomatic, afebrile, jaundiced infants younger than 8 weeks old . In addition, infants with the onset of jaundice after 8 days of age or patients with an elevated conjugated bilirubin fraction were more likely to have a UTI . Therefore, we recommend that testing for a UTI be included as part of the evaluation in asymptomatic, jaundiced infants presenting to the emergency department. Clin Diagn Lab Immunol, 2002 May, 9(3), 550 - 7 CD8 alpha-deficient mice are highly susceptible to 5-fluorouracil-induced lethality; Itoh N et al.; Intestinal intraepithelial lymphocytes (i-IEL) expressing CD8 alpha are located in the intestine and may confer protection against invasion of intestinal microflora . We found that mice rendered deficient in CD8 alpha molecules by homologous recombination were susceptible to 5-fluorouracil (5-FU)-induced lethality accompanied by translocation of members of the enterobacteria . The number of i-IEL was greatly reduced on day 6 after 5-FU administration in both CD8 alpha(+/-) mice and CD8 alpha(-/-) mice, whereas the recovery of the level of i-IEL thereafter was significantly impaired in CD8 alpha(-/-) mice compared with that in CD8 alpha(+/-) mice . The ability of i-IEL to produce gamma interferon in response to immobilized T-cell receptor (TCR) alpha beta or TCR gamma delta monoclonal antibodies was significantly lower in CD8 alpha(-/-) mice than in CD8 alpha(+/-) mice . Transfer of CD8(+) i-IEL conferred significant protection against 5-FU-induced lethality in CD8 alpha(-/-) mice . The results suggest that CD8(+) i-IEL play an important role in protection against 5-FU-induced lethality with translocation of Enterobacteriaceae. Mol Microbiol, 2002 Jan, 43(2), 307 - 22 Functional characterization of the antagonistic flagellar late regulators FliA and FlgM of Helicobacter pylori and their effects on the H . pylori transcriptome; Josenhans C et al.; Helicobacter pylori is thought to regulate gene expression with a very small set of regulatory genes . We identified a previously unannotated open reading frame (ORF) in the H . pylori 26695 genome (HP1122) as a putative H . pylori flgM gene (sigma28 factor antagonist) by a motif-based bioinformatic approach . Deletion of HP1122 resulted in a fourfold increase in transcription of the sigma28-dependent major flagellin gene flaA, supporting the function of HP1122 as H . pylori FlgM . Helicobacter pylori FlgM lacks a conserved 20-amino-acid N-terminal domain of enterobacterial FlgM proteins, but was able to interact with the Salmonella typhimurium sigma28 (FliA) and inhibit the expression of FliA-dependent genes in Salmonella . Helicobacter pylori FlgM inhibited FliA to the same extent in a Salmonella strain with an intact flagellar export system and in an export-deficient strain . Helicobacter pylori FliA was able to drive transcription of FliA-dependent genes in Salmonella . The effects of mutations in the H . pylori flgM and fliA genes on the H . pylori transcriptome were analysed using whole genome DNA microarrays . The antagonistic roles of FlgM and FliA in controlling the transcription of the major flagellin gene flaA were confirmed, and two additional FliA/FlgM dependent operons (HP472 and HP1051/HP1052) were identified . None of the three genes contained in these operons has a known function in flagellar biogenesis in other bacteria . Like other motile bacteria, H . pylori has a FliA/FlgM pair of sigma and anti-sigma factors, but the genes controlled by these differ markedly from the Salmonella/Escherichia coli paradigm. Drugs, 2002, 62(7), 1041 - 89 Ceftriaxone: an update of its use in the management of community-acquired and nosocomial infections; Lamb HM et al.; Ceftriaxone is a parenteral third-generation cephalosporin with a long elimination half-life which permits once-daily administration . It has good activity against Streptococcus pneumoniae, methicillin-susceptible staphylococci, Haemophilus influenzae, Moraxella catarrhalis and Neisseria spp . Although active against Enterobacteriaceae, the recent spread of derepressed mutants which hyperproduce chromosomal beta-lactamases and extended-spectrum beta-lactamases has diminished the activity of all third-generation cephalosporins against these pathogens necessitating careful attention to sensitivity studies . Extensive data from randomised clinical trials confirm the efficacy of ceftriaxone in serious and difficult-to-treat community-acquired infections including meningitis, pneumonia and nonresponsive acute otitis media . Ceftriaxone also has efficacy in other community-acquired infections including uncomplicated gonorrhoea, acute pyelonephritis and various infections in children . In the nosocomial setting, extensive data also confirm the efficacy of ceftriaxone with or without an aminoglycoside in serious Gram-negative infections, pneumonia, spontaneous bacterial peritonitis and as surgical prophylaxis . Outpatient use of ceftriaxone, either as part of a step-down regimen or parenterally, is a distinguishing feature of the data gathered on the agent over the last decade . The review focuses on new applica