J Food Prot, 2001 Nov, 64(11), 1756 - 60 Shellac formulations to reduce epiphytic survival of coliform bacteria on citrus fruit postharvest; McGuire RG et al.; Survival of the coliform bacteria Enterobacter aerogenes and Escherichia coli was monitored in a neutral carboxymethylcellulose formulation and in shellac formulations with various pH and concentrations of ethanol and the preservative paraben; populations were subsequently measured from the surface of citrus fruit coated with these formulations . Numbers of the two bacteria increased over 24 h from 10(6) CFU/ml to approximately 10(8) CFU/ml in the carboxymethylcellulose solution, but over this time numbers remained little changed in the neutral solution of shellac . The Enterobacter was more tolerant of alcohol over a 3-h period: although its numbers in a shellac solution with 10% ethanol dropped from more than 10(6) CFU/ml to just over 10(3) CFU/ml . E . coli and a third species . Klebsiella pneunoniae, declined toward the limit of detection (5 CFU/ ml) during this time . The addition of morpholine to increase the formulation pH to 9.0 caused numbers of bacteria to plummet to an undetectable level within 30 to 60 min . On Ruby Red grapefruit and Valencia oranges in storage at 13 degrees C numbers of E . aerogenes and E . coli declined over 2 weeks from 10(5) CFU/cm2 to less than 2.5 x 10(1), but most of the loss in numbers occurred within 1 day . Numbers remained significantly less on shellacked fruit compared with those applied in the carboxymethylcellulose coating, and a shellac coating prepared from a pH 9 solution was more toxic to these species than one in which 12% ethanol had been added to the neutral formulation.The addition of the preservative paraben in the basic shellac was further inhibitory. Tohoku J Exp Med, 2001 Aug, 194(4), 205 - 12 Enterobacterial repetitive intergenic consensus sequence-based PCR (ERIC-PCR); its ability to differentiate Streptococcus pyogenes strains and applicability to the study of outbreaks of streptococcal infection; Matsumoto M et al.; We evaluated the ability of enterobacterial repetitive intergenic consensus sequence-based PCR (ERIC-PCR) to differentiate 95 Streptococcus pyogenes strains with M or T serotypes isolated from sporadic streptococcal infections as compared with M or T serotypings and pulsed-field gel electrophoresis (PFGE) . Although the ERIC-PCR had less discriminatory power, defined as the ability to divide the strains with the same serotypes into the different sub-types, than PFGE, it consistently classified the strains into 16 patterns with a high correlation with M or T serotyping . The PCR method further discriminated 4 M or T serotypes into sub-types . The application of ERIC-PCR to 5 outbreaks of streptococcal infection produced the results that agreed closely with those of T serotyping and PFGE . ERIC-PCR has sufficient discriminatory power and is a quick and relatively easy technique, making it useful for routine epidemiological investigations. Transfusion, 2001 Nov, 41(11), 1356 - 64 Multiplex PCR for detection of Enterobacteriaceae in blood; Sen K et al.; BACKGROUND: Rapid and sensitive methods are needed to detect the small numbers of bacteria that may sometimes contaminate units of blood during collection . A multiplex 5'-nuclease TaqMan PCR assay (PE Applied Biosystems) was used to detect several bacterial species that may contaminate blood . STUDY DESIGN AND METHODS: Oligonucleotide primers were made for regions of the 16S rRNA gene conserved in four different bacterial species: Yersinia enterocolitica and Serratia, Klebsiella, and Enterobacter species . Two probes were designed: SL-1 detected Serratia, Klebsiella, and Enterobacter species, and YE-3 detected Y . enterocolitica . RESULTS: When TaqMan PCR was performed with chromosomal DNA isolated from pure cultures of Serratia liquefaciens, Klebsiella oxytoca, Klebsiella pneumoniae, Enterobacter cloacae, and Enterobacter agglomerans, the limit of detection with probe SL-1 was 1 to 2 CFUs . For S . marcescens, the sensitivity was 8 CFUs . The limit of detection for Y . enterocolitica with probe YE-3 was 2 CFUs . When total chromosomal DNA was extracted from whole-blood samples spiked with different numbers of Y . enterocolitica, S . liquefaciens, E . cloacae, or K . pneumoniae bacteria, the TaqMan PCR detected 12 to 16 organisms in 1 mL of blood . CONCLUSION: The 5'-nuclease TaqMan PCR assay takes only 3 hours to perform and has the potential to detect very small numbers of bacteria. Acta Microbiol Pol, 2001, 50(2), 161 - 7 Cloning of enterohemorrhagic Escherichia coli phage VT-2 dam methyltransferase; Radlinska M et al.; Enterobacterial GATC-specific DNA adenine methyltransferase (Dam) plays an essential role in regulation of DNA replication, methyl-directed mismatch repair, transposition and gene expression . In Salmonella typhimurium it has been shown to directly control virulence . In this paper we report cloning and expression of the dam gene from the Shiga toxin-producing VT2-Sa prophage of enterohemorrhagic Escherichia coli O157 . Comparisons of the predicted amino acid sequence indicates that Dam methyltransferases of E . coli phages VT2-Sa, 933W, T1 and Haemophilus influenzae phage HP1 make up a separate subgroup of adenine-N6 methyltransferases . These proteins are similar to the gamma subfamily of amino-methyltransferases in respect to the linear order of sequence motifs and the presence of the hallmark "NPPY" tetrapeptide . However, they apparently lack an autonomous target-recognizing domain at the C-terminus of the catalytic domain and therefore we propose to dub them as a "mini-gamma" subfamily. Pharmacotherapy, 2001 Aug, 21(8), 920 - 8 Extended-spectrum beta-lactamases: epidemiology, detection, and treatment; Nathisuwan S et al.; Extended-spectrum beta-lactamases (ESBLs) are extremely broad spectrum beta-lactamase enzymes found in a variety of Enterobacteriaceae . Most strains producing these beta-lactamases are Klebsiella pneumoniae, other Klebsiella species (i.e., K . oxytoca), and Escherichia coli . When producing these enzymes, organisms become highly effective at inactivating various beta-lactam antibiotics . In addition, ESBL-producing bacteria are frequently resistant to many classes of antibiotics, resulting in difficult-to-treat infections . Other problems due to ESBL-producing bacteria are difficulty in detecting the presence of ESBLs, limited treatment options, and deleterious impact on clinical outcomes . Clinicians should be familiar with the clinical significance of these enzymes and potential strategies for dealing with this growing problem. New Microbiol, 2001 Oct, 24(4), 365 - 9 Staphylococci in orthopaedic surgical wounds; Arciola CR et al.; From 50 infected surgical wounds of orthopaedic patients, 43 (86%) staphylococcal strains were isolated . 34 of all these staphylococci belonged to Staphylococcus aureus species (i.e . 68 % of all isolates from surgical wounds; 79% of staphylococcal isolates); 9 were coagulase-negative staphylococci (i.e . 21% of all isolates from surgical wounds; 18% of staphylococcal isolates) . Among microorganisms isolated from the wounds we also found 2 (4%) of the Enterobacteriaceae family; 2 (4%) of the Pseudomonas genus; 3 (6%) of the Streptococcus genus . Thus, orthopaedic surgical wounds were infected by staphylococci (mainly S . aureus) more frequently than by other micro-organisms . All the staphylococcal strains were screened for methicillin resistance by agar disk diffusion testing and for the presence of mecA gene responsible for methicillin resistance by PCR . 32% of the S . aureus and 33% of the S . epidermidis strains resulted methicillin resistant and mecA-positive . The data confirm the diffusion of methicillin resistant S . aureus in surgical site infections and shows that the so-called "new pathogens", i.e . S . epidermidis and other coagulase-negative staphylococci, also exhibit a frequent and hazardous methicillin-resisting ability. Proc Natl Acad Sci U S A, 2001 Dec 4, 98(25), 14607 - 12 Epub 2001 Nov 20. Mutation frequency and biological cost of antibiotic resistance in Helicobacter pylori; Bjorkholm B et al.; Among the several factors that affect the appearance and spread of acquired antibiotic resistance, the mutation frequency and the biological cost of resistance are of special importance . Measurements of the mutation frequency to rifampicin resistance in Helicobacter pylori strains isolated from dyspeptic patients showed that approximately 1/4 of the isolates had higher mutation frequencies than Enterobacteriaceae mismatch-repair defective mutants . This high mutation frequency could explain why resistance is so frequently acquired during antibiotic treatment of H . pylori infections . Inactivation of the mutS gene had no substantial effect on the mutation frequency, suggesting that MutS-dependent mismatch repair is absent in this bacterium . Furthermore, clarithromycin resistance conferred a biological cost, as measured by a decreased competitive ability of the resistant mutants in mice . In clinical isolates this cost could be reduced, indicating that compensation is a clinically relevant phenomenon that could act to stabilize resistant bacteria in a population. J Hosp Infect, 2001 Nov, 49(3), 183 - 92 A simultaneous outbreak on a neonatal unit of two strains of multiply antibiotic resistant Klebsiella pneumoniae controllable only by ward closure; Macrae MB et al.; Two aminoglycoside-resistant strains of Klebsiella pneumoniae caused an outbreak on the neonatal unit at St Thomas' Hospital . One, which affected 18 patients, was capsular type K18 and resistant to newer cephalosporins by the production of the extended-spectrum beta-lactamase SHV-2; the other, which colonized four patients, was capsular non-typeable and did not produce extended-spectrum beta-lactamase . Both strains were probably brought into the unit by carrier patients; the probable carrier of the non-typeable strain was transferred from another hospital but was negative on a single admission screen; the probable carrier of the K18 strain was not screened on admission because he had been born at St Thomas', but his mother had been transferred from another hospital . Despite intensive efforts to control the outbreak by standard methods of hand washing, screening, patient isolation and environmental cleaning, a total of 22 neonates on the unit eventually became colonized or infected . One of three patients with bacteraemia died . A small proportion of samples of expressed breast milk, electronic thermometers and oxygen saturation probes were contaminated by the K18 strain and may have contributed to some of the cross-infection, but this did not explain the extent of the outbreak . The outbreak was controlled only by opening a temporary ward for colonized neonates and another for newly born babies, which allowed the closure and cleaning of the main neonatal unit . Multiply antibiotic resistant klebsiellas may be highly epidemic and cause serious, difficult-to-control outbreaks on neonatal units . All patients, regardless of their admission history, should be screened on admission for carriage of multiply resistant enterobacteria by a sensitive method, and units should have plans for temporary ward closure should outbreaks occur . J Hosp Infect, 2001 Nov, 49(3), 173 - 82 Molecular epidemiological typing of Enterobacter cloacae isolates from a neonatal intensive care unit: three-year prospective study; Fernandez-Baca V et al.; Since 1992, there has been an increase in the incidence of Enterobacter sepsis in the neonatal intensive care unit (NICU) of the authors' hospital . From 1995 to 1997, a prospective molecular epidemiological survey of the colonizing and infecting strains isolated from neonates was conducted . Enterobacter cloacae was the most frequent cause of neonatal sepsis, accounting for 19.2% of all neonatal infections, reaching a peak incidence of 2.2/1000 during 1996 . Fifty isolates from the NICU and four epidemiologically unrelated strains were characterized by pulse-field gel electrophoresis (PFGE), ribotyping, enterobacterial repetitive intergenic consensus (ERIC)-PCR and plasmid profiling . PFGE was the most discriminatory technique and identified 13 types (two of them classified into two and three subtypes) compared with ERIC-PCR, plasmid profiling and ribotyping that identified 11, 11 and seven types, respectively . A good correlation was found between all techniques . Five different clones caused 15 cases of sepsis . Clones A and B were prevalent in 1995 and 1996, but they were not isolated in 1997 . An outbreak caused by clone G in 1997 was controlled by cohort nursing and hygienic measures, without changing the antibiotic policy . Strains were characterized by their antibiotic resistance pattern and divided into three groups . Group I correlated with PFGE types A, B1 and B2, which hyperproduced Bush type 1 chromosomal beta-lactamase and expressed extended-spectrum ?-lactamases (ESBLs) . Group II only hyperproduced Bush type 1 chromosomal beta-lactamase and correlated with PFGE-types D1, D2, D3 and I . Finally, Group III, with inducible beta-lactamases, correlated with the rest of PFGE types . The sudden disappearance of E . cloacae after reinforcement of hygienic measures confirms the importance of patient-to-patient transmission . Eur J Clin Microbiol Infect Dis, 2001 Sep, 20(9), 626 - 35 Multicenter evaluation of an automated system using selected bacteria that harbor challenging and clinically relevant mechanisms of resistance to antibiotics; Leclercq R et al.; A multicenter study was carried out to evaluate the performance of a new commercial automated system in comparison with that of the reference agar dilution method . Ten clinical microbiology laboratories tested a collection of 61 strains of gram-negative bacilli (49 Enterobacteriaceae and 12 Pseudomonas aeruginosa), and 6 other laboratories tested a collection of 55 strains of gram-positive cocci (10 enterococci and 45 staphylococci) against 10-20 antimicrobial agents . The strains were selected on the basis that they harbored challenging and characterized mechanisms of resistance . In comparison with the agar reference method, the automated system gave an overall essential agreement (+/-1 dilution) of 94.5%, 93.5%, and 97% for the gram-negative bacilli, enterococci, and staphylococci, respectively . According to the interpretive standards of the National Committee for Clinical Laboratory Standards, the category agreement ranged from 96 to 96.4% for the three sets of organisms . The accuracy of the automated system, as determined by the kappa test, ranged from 0.80 to 0.88, reflecting an almost perfect agreement with the reference technique . Very major, major, and minor errors obtained with the automated system were 0.3%, 2.9%, and 6.6% for gram-negative bacilli, 3.4%, 0%, and 5% for enterococci, and 1%, 1.6%, and 2.7% for staphylococci, respectively . The high rate of very major errors in enterococci was mostly due to a single strain of multidrug-resistant Enterococcus faecium, which was found susceptible to several antibiotics in a majority of participant laboratories . The use of a heavy inoculum and of a broth test medium by the automated system might account for a better expression of certain resistance mechanisms, including beta-lactamases, as compared to the agar dilution reference method . The interlaboratory reproducibility was acceptable, as shown by the narrow dispersion of MICs and by the results of quality control. Int J Antimicrob Agents, 2001 Nov, 18(5), 451 - 61 AS-924, a novel, orally active, bifunctional prodrug of ceftizoxime: physicochemical properties, oral absorption in animals, and antibacterial activity; Mori N et al.; AS-924 is an oral prodrug of the antibiotic ceftizoxime (CTIZ), a parenteral use cephalosporin . This novel prodrug, produced by esterifying CTIZ with a lipophilic pivaloyloxymethyl (POM) group and introducing a water soluble L-alanyl group, is expected to increase the bioavailability and thereby, augment the antibacterial activity of CTIZ in vivo compared with existing prodrugs . To study the effect of the L-alanyl group in AS-924 on its bioavailability, the plasma concentration profiles of CTIZ in dogs were examined following the dosing of AS-924 and CTIZ-POM, in powder form, after pretreatment with the antacid ranitidine, and following the dosing of AS-924 after pretreatment with a gastrointestinal motility stimulant metoclopramide or suppressant scopolamine butylbromide . The absorption rate of AS-924 was constant under these different conditions due to its unique balance of lipophilicity and water solubility . CTIZ is as antibacterially active as pre-existing oral cephalosporins against Gram-positive clinical isolates, while being more active against all Gram-negative isolates-particularly Enterobacteriaceae and Haemophilus influenzae . A simulation model for the eradication profile of bacteria in computer programmed pharmacokinetic (PK) system was carried out to study the antibacterial action of CTIZ in human . CTIZ was proven to eradicate Streptococcus pneumoniae and H . influenzae effectively, while cefpodoxime (CPOD), the active moiety of CPOD proxetil, eradicated S . pneumoniae, but not H . influenzae . These results confirm that, AS-924 is a potent oral antibiotic and would be expected to be clinically effective and efficient. Antimicrob Agents Chemother, 2001 Dec, 45(12), 3595 - 8 Characterization of a chromosomally encoded extended-spectrum class A beta-lactamase from Kluyvera cryocrescens; Decousser JW et al.; A chromosomally located beta-lactamase gene, cloned and expressed in Escherichia coli from a reference strain of the enterobacterial species Kluyvera cryocrescens, encoded a clavulanic acid-inhibited Ambler class A enzyme, KLUC-1, with a pI value of 7.4 . KLUC-1 shared 86% amino acid identity with a subgroup of plasmid-mediated CTX-M-type extended-spectrum beta-lactamases (CTX-M-1, -3, -10, -11, and -12), the most closely related enzymes, and 77% amino acid identity with KLUA-1 from Kluyvera ascorbata . The substrate profile of KLUC-1 corresponded to that of CTX-M-type enzymes. Microbiology, 2001 Nov, 147(Pt 11), 3105 - 11 Occurrence of two superoxide dismutases in Aeromonas hydrophila: molecular cloning and differential expression of the sodA and sodB genes; Leclere V et al.; Aeromonas spp., considered as emerging opportunistic pathogens, belong to the family Vibrionaceae . Among the criteria currently used for their classification is the presence of a single FeSOD (iron-containing superoxide dismutase), which distinguishes them from Enterobacteriacea . In this paper the cloning of the sodA and sodB genes encoding two different SODs in Aeromonas hydrophila ATCC 7966 is reported . The sodB gene encoded an FeSOD (196 amino acids, 21.5 kDa), was constitutively expressed and showed 75% homology with the E . coli FeSOD . The sodA gene encoded a protein of 206 amino acids (22.5 kDa) with MnSOD (manganese-containing SOD) activity and showed 55% homology with the Escherichia coli MnSOD . The MnSOD of A . hydrophila was detected only during the stationary phase of growth under high aeration or when induced by lack of iron . Nevertheless, paraquat had no detectable effect on its production . The amino-terminal part of the Mn-containing protein contained a putative signal sequence which could permit a periplasmic localization. Microbiology, 2001 Nov, 147(Pt 11), 3015 - 25 Conserved amino acid residues found in a predicted cytosolic domain of the lipopolysaccharide biosynthetic protein WecA are implicated in the recognition of UDP-N-acetylglucosamine; Amer AO et al.; WecA, an integral membrane protein that belongs to a family of polyisoprenyl phosphate N-acetylhexosamine-1-phosphate transferases, is required for the biosynthesis of O-specific LPS and enterobacterial common antigen in Escherichia coli and other enteric bacteria . WecA functions as an UDP-N-acetylglucosamine (GlcNAc):undecaprenyl-phosphate GlcNAc-1-phosphate transferase . A conserved short sequence motif (His-Ile-His-His; HIHH) and a conserved arginine were identified in WecA at positions 279-282 and 265, respectively . This region is located within a predicted cytosolic segment common to all bacterial homologues of WecA . Both HIHH279-282 and the Arg265 are reminiscent of the HIGH motif (His-Ile-Gly-His) and a nearby upstream lysine, which contribute to the three-dimensional architecture of the nucleotide-binding site among various enzymes displaying nucleotidyltransferase activity . Thus, it was hypothesized that these residues may play a role in the interaction of WecA with UDP-GlcNAc . Replacement of the entire HIHH motif by site-directed mutagenesis produced a protein that, when expressed in the E . coli wecA mutant MV501, did not complement the synthesis of O7 LPS . Membrane extracts containing the mutated protein failed to transfer UDP-GlcNAc into a lipid-rich fraction and to bind the UDP-GlcNAc analogue tunicamycin . Similar results were obtained by individually replacing the first histidine (H279) of the HIHH motif as well as the Arg265 residue . The functional importance of these residues is underscored by the high level of conservation of H279 and Arg265 among bacterial WecA homologues that utilize several different UDP-N-acetylhexosamine substrates. Microbiology, 2001 Nov, 147(Pt 11), 2951 - 9 Molecular comparison of pathogenic bacteria from pear trees in Japan and the fire blight pathogen Erwinia amylovora; Kim WS et al.; Several strains of the genus Erwinia, which were isolated in Japan from pear trees with necrotic symptoms that resembled fire blight, and tentatively identified as Erwinia amylovora, were reinvestigated for their relationship to the fire blight pathogen . These isolates produced ooze on slices of immature pears and were mucoid on MM2Cu agar plates, but did not synthesize levan and did not give the expected PCR signals with several primer pairs specific for Erwinia amylovora . The isolates tested positive with PCR primers designed to detect the novel pear pathogen Erwinia pyrifoliae, which was isolated from Nashi pear trees in South Korea . The nucleotide sequence analysis of a DNA fragment preceding the gene cluster for exopolysaccharide synthesis revealed a closer relationship to Erwinia pyrifoliae than to Erwinia amylovora . Plasmid profiles, protein patterns and genomic DNA analysed by PFGE after XbaI and SpeI digestion were different than Erwinia amylovora . Experiments with strains of Erwinia amylovora isolated from raspberry (Rubus sp.), Erwinia mallotivora and Enterobacter pyrinus also did not reveal a relationship between these bacteria and the Japanese Erwinia strains . The latter are not identical to Erwinia pyrifoliae, but possess many similar features to this pathogen that causes Asian pear blight . It is concluded that pathogenic bacteria isolated in Japan from pear trees with symptoms resembling fire blight are possibly different from Erwinia amylovora. J Bioenerg Biomembr, 2001 Jun, 33(3), 179 - 86 The Na+-translocating NADH:quinone oxidoreductase (NDH I) from Klebsiella pneumoniae and Escherichia coli: implications for the mechanism of redox-driven cation translocation by complex I; Steuber J; Eukaryotic complex I integrated into the respiratory chain transports at least 4 H+ per NADH oxidized . Recent results indicate that the cation selectivity is altered to Na+ in complex I (NDH I) isolated from the enterobacteria Escherichia coli and Klebsiella pneumoniae . A sequence analysis illustrates the characteristic differences of the enterobacterial, Na+-translocating NDH I compared to the H+-translocating complex I from mitochondria . Special attention is given to the membranous NuoL (ND5, Nqo12) subunits that possess striking sequence similarities to secondary Na+/H+ antiporters and are proposed to participate in Na+ transport . A model of redox-linked Na+ (or H+) transport by complex I is discussed based on the ion-pair formation of a negatively charged ubisemiquinone anion with a positively charged Na+ (or H+). Ther Umsch, 2001 Oct, 58(10), 609 - 13 {Nosocomial pneumonia}; Ewig S et al.; Nosocomial pneumonia is a frequent complication, particularly during mechanical ventilation . Microbial patterns differ according to the time of onset: whereas early onset pneumonia (up to the fourth day of hospitalization) is mostly caused by Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae, leading pathogens of late onset pneumonia (after the fourth day) additionally include gram-negative Enterobacteriaceae and potentially drug resistant microorganisms . These general patterns are modified in the presence of specific individual risk factors . The diagnostic work-up of pneumonia is aimed at the assessment of severity, the confirmation of the presence of pneumonia as well as the identification of the causal pathogens . The results of microbiological investigations must always be interpreted in the clinical context . Antimicrobial treatment must always be initiated empirically based on expected microbial patterns as outlined above . Nosocomial pneumonia in nonventilated and ventilated patients requires different empirical antimicrobial treatment approaches . All patients with antimicrobial treatment failures must be comprehensively reevaluated. Int J Antimicrob Agents, 2001 Oct, 18(4), 365 - 71 Detection of amp C in Enterobacter cloacae in China; Zhang YL et al.; PCR amplification of 55 strains of Enterobacter cloacae indicated 51 of them had amp C structural gene verified by DNA sequence and Southern blotting . All PCR products were cleaved into 666- and 328-bp fragments by Kpn1 restriction enzyme . Imipenem was the most potent inducer for mRNA expression of amp C gene and beta-lactamase activity . The beta-Lactamase inhibitor R0481220 strongly inhibited Amp C beta-lactamases; 96.4% (53/55) of Enterobacter cloacae producing Amp C enzyme were susceptible to cefepime. J Med Chem, 2001 Nov 8, 44(23), 4023 - 6 New pyridoquinoline derivatives as potential inhibitors of the fluoroquinolone efflux pump in resistant Enterobacter aerogenes strains; Chevalier J et al.; Enterobacter aerogenes, one of the most frequently isolated nosocomial pathogens in France, is exhibiting increasing multidrug resistance mechanisms associated with a change in membrane permeability . For drugs of the quinolone family, mutations in the target and active efflux play a prominent role in the resistance . We report here the effect of several pyridoquinoline derivatives that restore a noticeable fluoroquinolone accumulation to resistant strains that overexpress the MarA activator . Studies of the energy-dependent quinolone efflux indicate that the most efficient derivatives tested probably inhibit the resistance process by acting as substrate competitors on the pump extruding intracellular norfloxacin. J Ind Microbiol Biotechnol, 2001 Oct, 27(4), 228 - 33 Bacterial amelioration of bauxite residue waste of industrial alumina plants; Hamdy MK et al.; The high alkali content of bauxite residue deposits from alumina production plants in industrial nations poses a challenge to reestablish flora and fauna at the deposit sites . The present study demonstrated that low levels of injured bacterial cells in the bauxite residue actively grew using various added nutrients and/or hay . The organisms grew from less than 10 to more than 10(9) cells g(-1) bauxite residue and formed organic acids that lowered the pH from 13 to about 7.0 . A total of 150 cultures was isolated from treated bauxite residue and included species of Bacillus, Lactobacillus, Leuconostoc, Micrococcus, Staphylococcus, Pseudomonas, Flavobacterium and Enterobacter . Scanning electron micrographs demonstrated that untreated particles (control) of the bauxite residue were clumped together, and in treated bauxite residue these particles were highly dispersed with microcolonial structures . Furthermore, the treated bauxite residue supported growth of several plants and earthworms that survived for over 300 days . In a test plot bioremediation on a residue deposit at Alcoa Point Comfort, TX, the Bermuda grass hay used was effective mulch material and encouraged water filtration, leading to establishment and growth of salt-tolerant vegetative species. Diagn Microbiol Infect Dis, 2001 Sep-Oct, 41(1-2), 65 - 70 Validation of the VITEK2 and the Advance Expert System with a collection of Enterobacteriaceae harboring extended spectrum or inhibitor resistant beta-lactamases; Canton R et al.; The susceptibility testing accuracy of the VITEK2 system and the ability of the Advance Expert System (AES) to provide interpretive readings were evaluated against 86 extended spectrum (ESBL) and 6 inhibitor-resistant-TEM (IRT) beta-lactamases producing Enterobacteriaceae clinical isolates . VITEK2 MICs of 12 beta-lactams were compared with those obtained by the standard NCCLS microdilution technique . The overall essential agreement ( +/- 1 log dilution) was 87.8% . Discrepancies were mainly observed with cefepime (30.3% of total number of discrepancies), ceftazidime (21.2%), and cefotaxime (15.1%) . MIC discrepancies were slightly higher in CTX-M- (14.4%) than in TEM- (12.5%) or SHV- (11.9%) type ESBL producers and were rare in IRT producers (1.4%) . Overall interpretive agreement was 92.5% and minor, major, and very major errors were 5.4%, 1.7%, and 2.1%, respectively . The AES was able to identify an ESBL phenotype in 85 out of 86 isolates (98.8%) and an IRT phenotype in all 6 isolates harboring these enzymes, thus reducing very major errors to 0.9% . The VITEK2 system, in conjunction with the AES software, is a reliable tool for detection of ESBL or IRT producing Enterobacteriaceae isolates. Diagn Microbiol Infect Dis, 2001 Sep-Oct, 41(1-2), 1 - 14 Cefditoren in vitro activity and spectrum: a review of international studies using reference methods; Jones RN et al.; Cefditoren, a broad-spectrum orally administered cephalosporin ester, has documented in vitro efficacy against many Gram-positive and -negative pathogens and stability against clinically important beta-lactamases . We have reviewed the microbiology and the pharmacokinetic/pharmacodynamic literature regarding the spectrum and potency of this newer agent against the major etiologic agents of community-acquired respiratory infection, (Streptococcus pneumoniae, Hemophilus influenzae and Moraxella catarrhalis), as well as the Enterobacteriaceae and non-enteric Gram-negative bacilli, staphylococci, and other aerobic and anaerobic Gram-positive cocci . The level of cefditoren activity against S . pneumoniae (MIC(90,) 0.5 microg/mL) was superior to all marketed oral cephalosporins and at least equal to amoxicillin +/- clavulanate . H . influenzae (MIC(90,) 0.016-0.03 microg/mL) and M . catarrhalis (MIC(90,) 0.06-0.5 microg/mL) were also very susceptible to cefditoren . In contrast to cefixime and ceftibuten, cefditoren was active against oxacillin-susceptible staphylococci (MIC(90,) < or = 1 microg/mL) at a level comparable to cefuroxime axetil, cefaclor or cefprozil . Enterococci, Pseudomonas aeruginosa and most anaerobes (Gram-negative) were not cefditoren-susceptible, but most Enterobacteriaceae, beta-haemolytic and viridans group streptococci were highly susceptible . Furthermore, an overview of key in vitro susceptibility testing methods and issues including disk diffusion testing and Etest (AB BIODISK, Solna, Sweden) method accuracy, interpretive criteria, and pharmacodynamic considerations for the selection of a breakpoint concentration are provided . The rapid bactericidal nature of the antibacterial activity of cefditoren, its post antibiotic effect, penicillin binding protein targets, and extent of beta-lactamase stability are all favorable qualities . In conclusion, this orally administered (BID) beta-lactam possesses promise for use against commonly isolated problematic respiratory tract pathogens such as penicillin-non-susceptible pneumococci and beta-lactamase-positive M . catarrhalis or H . influenzae . Success in the clinical trials will further define the role of cefditoren in this era of emerging resistant bacterial pathogens. J Gastroenterol Hepatol, 2001 Oct, 16(10), 1112 - 9 Protective effect of rebamipide on indomethacin-induced intestinal damage in rats; Mizoguchi H et al.; BACKGROUND AND AIM: We evaluated the effect of rebamipide (2-(4-chlorobenzoylamino)-3-{2(1H)-quinolinon-4-yl} propionic acid), a novel anti-ulcer drug, on indomethacin-induced small intestinal lesions in rats . METHODS: The animals were administered indomethacin (10 mg/kg, s.c.), and they were killed 24 h later . Rebamipide (30-300 mg/kg) was administered p.o . twice, 30 min before, and 6 h after indomethacin . RESULTS: Indomethacin caused hemorrhagic lesions in the rat small intestine, accompanied by an increase in enterobacterial translocation, inducible nitric oxide synthase (iNOS) and myeloperoxidase (MPO) activities, as well as thiobarbituric acid (TBA) reactants, and these changes were significantly prevented by the supplementation with 16,16-dimethyl prostaglandin E2 (dmPGE2; 10 microg/kg, i.v.) or the pretreatment of animals with the antibiotic ampicillin . Treatment of the animals with rebamipide dose-dependently prevented the development of intestinal lesions, and this effect was mimicked by i.v . administration of superoxide dismutase (SOD: 3000 U/kg) + catalase (CAT: 5000 U/kg) . The protection by rebamipide was accompanied by a significant suppression of the increase in both MPO and iNOS activities, and a complete inhibition of the increase in TBA reactants, while SOD + CAT significantly inhibited the increase of MPO activity and TBA reactants, but not iNOS activity . The bacterial translocation following indomethacin was also significantly decreased by either rebamipide or SOD + CAT . CONCLUSION: These results confirmed the importance of enterobacteria and iNOS/NO in the pathogenesis of indomethacin-induced small intestinal lesions, and suggested that rebamipide prevents the development of these lesions, probably by its radical scavenging action. Curr Microbiol, 2001 Dec, 43(6), 448 - 51 Recombinant Enterobacter amnigenus highly toxic to Anopheles dirus mosquito larvae; Khampang P et al.; The mosquito-larvicidal binary toxin of Bacillus sphaericus 2297 was expressed in Enterobacter amnigenus, a Gram-negative bacterium isolated from Anopheles dirus larvae gut . The toxin was placed under the regulation of various promoters in order to improve the expression level of the toxin . Amongst the recombinants obtained, E . amnigenus harboring pBS373, a plasmid which contains the toxin genes under the control of the native B . sphaericus promoter, expressed a significant amount of protein, comparable to that found in B . sphaericus 2297 . In addition, this recombinant provided approximately twenty times higher toxicity against second-instar Anopheles dirus larvae when compared to B . sphaericus 2297 . The procedure of obtaining this environmentally isolated bacterium from larvae gut and introducing the system for mosquito-larvicidal toxin synthesis is noteworthy . The promising result presented here provides a substantial degree of confidence for further field studies. Clin Microbiol Infect, 2001 Oct, 7(10), 553 - 61 Trends in quinolone susceptibility of Enterobacteriaceae among inpatients of a large university hospital: 1992-98; Robert J et al.; OBJECTIVES: To assess trends in quinolone susceptibility of Enterobacteriaceae isolated in a large university hospital . METHODS: Between 1992 and 1998, bacterial isolates were collected each year during a 3-month period to evaluate annual changes in susceptibility . In addition, the activities of fluoroquinolones (pefloxacin, norfloxacin, ofloxacin, ciprofloxacin) against nalidixic acid-resistant strains were determined by disk diffusion and MIC methodologies during the first and last year of the study . RESULTS: The susceptibility of Enterobacteriaceae to nalidixic acid was unchanged between 1992 and 1998 (86% versus 85%) . However, at the species level, the susceptibility rates to nalidixic acid decreased for Escherichia coli from 92% to 89%, and for Enterobacter cloacae from 87% to 82% . In contrast, there was a 10% increase in the nalidixic acid susceptibility rates for Klebsiella pneumoniae (74% versus 83%), which was thought to be due to the control of the spread of epidemic extended-spectrum beta-lactamase (ESBL)-producing strains . The overall susceptibility of the Enterobacteriaceae to the fluoroquinolones remained high during the study period, greater than 90% in the case of ciprofloxacin . However, nalidixic acid-resistant Escherichia coli showed decreased susceptibility to ciprofloxacin between 1992 and 1998, as reflected by a decrease in median zone diameter (26 mm to 19 mm), an increase in MIC(50) (0.25 mg/L to 1 mg/L) and a shift in MIC distribution (unimodal in 1992 to bimodal in 1998) . This has resulted in the reduced susceptibility of Escherichia coli to fluoroquinolones between 1992 and 1998 (pefloxacin, 95-90%; ciprofloxacin, 99-95%) . CONCLUSIONS: The susceptibility of Escherichia coli to quinolones has decreased, and the level of susceptibility of the resistant strains has increased over the 7-year study period. J Clin Microbiol, 2001 Nov, 39(11), 3865 - 70 PCR analyses of tRNA intergenic spacer, 16S-23S internal transcribed spacer, and randomly amplified polymorphic DNA reveal inter- and intraspecific relationships of Enterobacter cloacae strains; Clementino MM et al.; PCR analysis of tRNA intergenic spacer (tDNA-PCR) and of the 16S-23S internal transcribed spacer (ITS-PCR) and random amplified polymorphic DNA (RAPD) analysis were evaluated for their usefulness in characterization of Enterobacter cloacae strains isolated from both clinical origins and vaccine microbial contamination . tDNA-PCR presented specific and reproducible patterns for Enterobacter sakazakii ATCC 29004, Enterobacter aerogenes ATCC 13048, and Enterobacter cloacae ATCC 13047 and 23355 that presented the same profile for all 16 E . cloacae isolates, offering an alternative tool for species-level identification . ITS-PCR and RAPD analysis yielded completely different banding patterns for the 20 strains studied, except for E . cloacae strains isolated from different batches of vaccine that exhibited a unique pattern, suggesting contamination by the same strain . The combined use of tDNA-PCR and ITS-PCR in a one-step protocol allows accurate identification and typing of E . cloacae strains a few hours after the colony has been isolated. Appl Environ Microbiol, 2001 Nov, 67(11), 5285 - 93 Endophytic colonization and in planta nitrogen fixation by a Herbaspirillum sp . isolated from wild rice species; Elbeltagy A et al.; Nitrogen-fixing bacteria were isolated from the stems of wild and cultivated rice on a modified Rennie medium . Based on 16S ribosomal DNA (rDNA) sequences, the diazotrophic isolates were phylogenetically close to four genera: Herbaspirillum, Ideonella, Enterobacter, and Azospirillum . Phenotypic properties and signature sequences of 16S rDNA indicated that three isolates (B65, B501, and B512) belong to the Herbaspirillum genus . To examine whether Herbaspirillum sp . strain B501 isolated from wild rice, Oryza officinalis, endophytically colonizes rice plants, the gfp gene encoding green fluorescent protein (GFP) was introduced into the bacteria . Observations by fluorescence stereomicroscopy showed that the GFP-tagged bacteria colonized shoots and seeds of aseptically grown seedlings of the original wild rice after inoculation of the seeds . Conversely, for cultivated rice Oryza sativa, no GFP fluorescence was observed for shoots and only weak signals were observed for seeds . Observations by fluorescence and electron microscopy revealed that Herbaspirillum sp . strain B501 colonized mainly intercellular spaces in the leaves of wild rice . Colony counts of surface-sterilized rice seedlings inoculated with the GFP-tagged bacteria indicated significantly more bacterial populations inside the original wild rice than in cultivated rice varieties . Moreover, after bacterial inoculation, in planta nitrogen fixation in young seedlings of wild rice, O . officinalis, was detected by the acetylene reduction and (15)N(2) gas incorporation assays . Therefore, we conclude that Herbaspirillum sp . strain B501 is a diazotrophic endophyte compatible with wild rice, particularly O . officinalis. Clin Microbiol Infect, 2001 Sep, 7(9), 470 - 8 The changing nature of aminoglycoside resistance mechanisms and prevalence of newly recognized resistance mechanisms in Turkey; Over U et al.; OBJECTIVE: To determine the most frequently occurring individual and combined resistance mechanisms in Gram-negative bacteria resistant to any of the clinically available aminoglycosides in Turkey, and to compare these mechanisms with those found in smaller, earlier studies . METHODS: Aminoglycoside resistance mechanisms in Gram-negative isolates resistant to either gentamicin, tobramycin, netilmicin or amikacin collected in different regions of Turkey were evaluated both phenotypically and genotypically using 12 aminoglycosides and up to 22 aminoglycoside resistance gene probes . RESULTS: Among 696 aminoglycoside-resistant Gram-negative bacteria, resistance rates were very high for gentamicin (94.5%), tobramycin (82.4%), netilmicin (53.6%), and amikacin (49.7%) . Although isepamicin was the most active aminoglycoside against Gram-negative bacteria, increased resistance (29.7%) was found and resistance rates were higher than those in most of the other countries surveyed in earlier studies . The most common aminoglycoside resistance mechanisms (AAC(3)-II (GTN), AAC(6')-I (TNA), and ANT(2")-I (GT)) in the earlier studies were also found in the present isolates of Klebsiella spp., Enterobacter spp . and Escherichia coli, with increased complexity . In addition to these old mechanisms, two new aminoglycoside resistance mechanisms, namely AAC(6')-III (TNAI) and AAC(6')-IV (GTNA), were also found at significant frequencies (11.9% and 26.9%, respectively) in these isolates of Enterobacteriaceae (n = 435) . Among the isolates of Pseudomonas spp . (n = 150), in addition to the increased complexity of enzymatic resistance mechanisms (AAC(3)-I (16.6%), AAC(6')-II (29.3%), AAC(6')-III (19.3%), ANT(2")-I (40%)), permeability resistance seemed to be responsible for the high rates of resistance to aminoglycosides . CONCLUSION: The results of this study indicated increased resistance to clinically available aminoglycosides, including isepamicin, even though it was the most active, as a result of both the presence of new aminoglycoside resistance mechanisms and the increased complexity of all mechanisms, including permeability resistance, particularly in Pseudomonas in Turkey. Arch Latinoam Nutr, 2001 Jun, 51(2), 173 - 9 {Microbiological stability study of minimally processed cantaloupe (Cucumis melo L) by vacuum}; Millan FR et al.; In order to design a minimal process for cantaloupe, the water activity of the fruit was reduced until 0.976 through vacuum osmotic dehydration and the optimum combination of microbial preservatives (100 ppm sodium sulfite, 600 ppm potassium sorbate, 0.5% L-ascorbic acid and 1% citric acid), that contributed to minimize (p < 0.05), the content of aerobic mesophilic bacteria, molds and yeast, was determined through a 2k multifactorial design . In the second experimental stage, the conditions previously mentioned were tested together with the application of fruit's blanching (98 degrees C/3 min) with saturated steam and product storage in three different modified atmospheres (6% O2-6% CO2, 5% O2-10% CO2 and atmospheric air) . In such sense, the application of microbial preservatives together with fruit's blanching and atmospheric air in high barrier packages (30 mu nylon) contributed to extent the shelf life of the cantaloupe minimally processed during 14 days on the basis of aerobic mesophilic bacteria, psychrotrophic bacteria, acid lactic bacteria, enterobacteria, molds, yeast and the sensory attributes color, odour, taste and texture. Cas Lek Cesk, 2001 Aug, 140(16), 487 - 91 {New aspects of antibiotic resistance and possibilities of its prevention}; Blahova J et al.; New phenomena of the antibiotic resistance in bacteria have recently appeared . The may hold present explosive development of resistance and prevent its transferability from multiple drug resistant bacteria to still sensitive ones . They may prevent the production of so-called extended-spectrum beta-lactamases (ESBLs) among Enterobacteriaceae producing resistance virtually to all penicillins and cephalosporins with exception of those antibiotics potentiated by clavulanic acid or sulbactam, the resistance to vancomycin in enterococci and staphylococce, and the resistance of Stenotrophomonas maltophilia . Factors participating on the development of resistance include: a) transferability of resistance genes among bacteria which explosively change susceptible strains to resistant ones, b) dosage and types of antibiotics which cause the selection pressure to certain species of bacteria, c) level of organization and strict adherence to hygienic and anti-epidemic regimen starting with the entry of patients into the hospital . Analyses are necessary to check whether the patient brings resistant bacteria with a transferable resistance (with ESBLs) into the hospital . Preventive measures would be strictly applied to stop the clonal spread of resistant strains among the patients and/or hospital environment, which occurs if these strains have such opportunity . Last, but not least to be considered is the dosage, composition and rationality of administration of antibacterials, mainly in post-operative prophylaxis in intensive care units, in so-called empirical usage, etc . At the same time, it would be highly unethical to hesitate with application of antibacterials to patients when it is justified, necessary and rational . Hospital antibiotics policy should rationally decide between these alternatives in each application of antibiotics or their combinations. J Formos Med Assoc, 2001 Aug, 100(8), 548 - 52 Comparison of ofloxacin and norfloxacin concentration in prostatic tissues in patients undergoing transurethral resection of the prostate; Chen J et al.; BACKGROUND AND PURPOSE: To compare the concentrations of two fluoroquinolones, ofloxacin (OFLX) and norfloxacin (NFLX), in the prostate glands of patients who underwent transurethral resection of the prostate (TUR-P) after oral ingestion of both drugs for surgical prophylaxis . METHODS: Ten patients with clinical symptoms of benign prostatic hyperplasia undergoing TUR-P received 200 mg of both drugs per os simultaneously 2 hours before surgery . The concentrations of the drugs in the serum and prostate at the time of surgery were measured by high performance liquid chromatography . Patients' clinical characteristics were evaluated, including findings from transrectal ultrasonography of the prostate, prostate specific antigen concentration, renal function tests, and post-operative status . RESULTS: Two hours after administration, the mean serum concentration of OFLX was 4.14 +/- 0.64 mg/L (range 0.27-6.37) and of NFLX was 1.10 +/- 0.22 mg/L (range 0.02-2.1) . The concentration of ORLX in prostatic tissue was 4.10 +/- 0.79 micrograms/g (range 1.70-6.37) and of NFLX was 2.22 +/- 0.57 micrograms/g (range 0.63-4.35) . The ratio of the prostatic tissue concentration (P) to the serum concentration (S) was 2.11 for OFLX and 5.71 for NFLX . The concentrations of both drugs exceeded the minimum inhibitory concentration (MIC) for most gram-negative organisms, but NFLX may be unable to exceed the MIC90 of Enterobacter cloacae in some individuals . Leukocytosis and spiking fever developed after TUR-P in two of the 10 patients . CONCLUSIONS: Concentrations of OFLX were higher in both serum and prostatic adenoma tissues than those of NFLX (p < 0.001), while NFLX had better penetration into the prostate (P/S ratio) (p < 0.001) . The results of this study indicated that the concentrations of both of these drugs exceeded the MIC for most gram-negative organisms. Nature, 2001 Oct 25, 413(6858), 852 - 6 Complete genome sequence of Salmonella enterica serovar Typhimurium LT2; McClelland M et al.; Salmonella enterica subspecies I, serovar Typhimurium (S . typhimurium), is a leading cause of human gastroenteritis, and is used as a mouse model of human typhoid fever . The incidence of non-typhoid salmonellosis is increasing worldwide, causing millions of infections and many deaths in the human population each year . Here we sequenced the 4,857-kilobase (kb) chromosome and 94-kb virulence plasmid of S . typhimurium strain LT2 . The distribution of close homologues of S . typhimurium LT2 genes in eight related enterobacteria was determined using previously completed genomes of three related bacteria, sample sequencing of both S . enterica serovar Paratyphi A (S . paratyphi A) and Klebsiella pneumoniae, and hybridization of three unsequenced genomes to a microarray of S . typhimurium LT2 genes . Lateral transfer of genes is frequent, with 11% of the S . typhimurium LT2 genes missing from S . enterica serovar Typhi (S . typhi), and 29% missing from Escherichia coli K12 . The 352 gene homologues of S . typhimurium LT2 confined to subspecies I of S . enterica-containing most mammalian and bird pathogens-are useful for studies of epidemiology, host specificity and pathogenesis . Most of these homologues were previously unknown, and 50 may be exported to the periplasm or outer membrane, rendering them accessible as therapeutic or vaccine targets. J Am Chem Soc, 2001 Oct 31, 123(43), 10436 - 43 Mechanism of inhibition of the class C beta-lactamase of Enterobacter cloacae P99 by cyclic acyl phosph(on)ates: rescue by return; Kaur K et al.; As previously described (Pratt, R . F.; Hammar, N . J . J . Am . Chem . Soc . 1998, 120, 3004.), 1-hydroxy-4,5-benzo-2,6-dioxaphosphorinone(3)-1-oxide (salicyloyl cyclic phosphate) inactivates the class C beta-lactamase of Enterobacter cloacae P99 in a covalent fashion . The inactivated enzyme slowly reverts to the active form . This paper shows that reactivation involves a recyclization reaction that regenerates salicyloyl cyclic phosphate rather than hydrolysis of the covalent intermediate . The inactivation, therefore, is a slowly reversible covalent modification of the active site . The thermodynamic dissociation constant of the inhibitor from the inactivated enzyme is 0.16 microM . Treatment of the inactivated enzyme with alkali does not produce salicylic acid but does, after subsequent acid hydrolysis, yield one molar equivalent of lysinoalanine . This result proves that salicyloyl cyclic phosphate inactivates the enzyme by (slowly reversible) phosphorylation of the active site serine residue . This result contrasts sharply with the behavior of acyclic acyl phosphates which transiently inactivate the P99 beta-lactamase by acylation (Li, N.; Pratt, R . F . J . Am . Chem . Soc . 1998, 120, 4264.) . This chemoselectivity difference is explored by means of molecular modeling . Rather counterintuitively, in view of the relative susceptibility of phosphates and phosphonates to nucleophilic attack at phosphorus, 1-hydroxy-4,5-benzo-2-oxaphosphorinanone(3)-1-oxide, the phosphonate analogue of salicyloyl cyclic phosphate, did not appear to inactivate the P99 beta-lactamase in a time-dependent fashion . It was found, however, to act as a fast reversible inhibitor (K(i) = 10 microM) . A closer examination of the kinetics of inhibition revealed that both on and off rates (9.8 x 10(3) s(-1) x M(-1) and 0.098 s(-1), respectively) were much slower than expected for noncovalent binding . This result strongly indicates that the inhibition reaction of the phosphonate also involves phosphylation of the active site . Hence, unlike the situation with bacterial DD-peptidases covalently inactivated by beta-lactams, the P99 beta-lactamase inactivated by the above cyclic acyl phosph(on)ates can be rescued by return . Elimination of the recyclization reaction would lead to more effective inhibitors. J Immunol, 2001 Nov 1, 167(9), 5278 - 85 Differential induction of endotoxin tolerance by lipopolysaccharides derived from Porphyromonas gingivalis and Escherichia coli; Martin M et al.; Exposure of mononuclear phagocytes to enterobacterial LPS induces a state of transient hyporesponsiveness to subsequent LPS exposure, termed endotoxin tolerance . In the present study, LPS derived from the oral periodontal pathogen, Porphyromonas gingivalis, was compared with that derived from the enterobacterium, Escherichia coli, for the ability to induce endotoxin tolerance . Pretreatment of the human macrophage cell line, THP-1, with E . coli LPS resulted in a severe reduction in the levels of IL-1beta, IL-6, and TNF-alpha upon secondary stimulation . In contrast, pretreatment of THP-1 cells with P . gingivalis LPS resulted in a mitigation of IL-1beta, but not IL-6 and TNF-alpha production upon subsequent exposure to P . gingivalis LPS: primary or secondary stimulation with < or =100 ng/ml P . gingivalis LPS resulted in comparable levels of IL-6 and TNF-alpha, while stimulation of THP-1 cells with > or =1 microg/ml P . gingivalis LPS induced a significant enhancement in IL-6 and TNF-alpha levels upon secondary exposure . To identify possible mechanisms for these differences, changes in the expression of molecules involved in the LPS-signaling pathway were assessed . Pretreatment of THP-1 cells with E . coli LPS resulted in a significant reduction in surface Toll-like receptor 4 (TLR4) expression and an inability to degrade I-kappaB-alpha or I-kappaB-beta proteins upon secondary stimulation . In contrast, pretreatment of THP-1 cells with P . gingivalis LPS resulted in a significant enhancement of both CD14 and TLR2, while maintaining the ability to degrade I-kappaB-beta only upon secondary stimulation . Thus, E . coli and P . gingivalis LPS differentially affect CD14 and TLR expression as well as secondary LPS-associated responses. J Bacteriol, 2001 Nov, 183(22), 6509 - 16 Identification of the structural gene for the TDP-Fuc4NAc:lipid II Fuc4NAc transferase involved in synthesis of enterobacterial common antigen in Escherichia coli K-12; Rahman A et al.; The polysaccharide chains of enterobacterial common antigen (ECA) are comprised of the trisaccharide repeat unit Fuc4NAc-ManNAcA-GlcNAc, where Fuc4NAc is 4-acetamido-4,6-dideoxy-D-galactose, ManNAcA is N-acetyl-D-mannosaminuronic acid, and GlcNAc is N-acetyl-D-glucosamine . Individual trisaccharide repeat units are assembled as undecaprenyl-linked intermediates in a sequence of reactions that culminate in the transfer of Fuc4NAc from TDP-Fuc4NAc to ManNAcA-GlcNAc-pyrophosphorylundecaprenol (lipid II) to yield Fuc4NAc-ManNAcA-GlcNAc-pyrophosphorylundecaprenol (lipid III), the donor of trisaccharide repeat units for ECA polysaccharide chain elongation . Most of the genes known to be involved in ECA assembly are located in the wec gene cluster located at ca . 85.4 min on the Escherichia coli chromosome . The available data suggest that the structural gene for the TDP-Fuc4NAc:lipid II Fuc4NAc transferase also resides in the wec gene cluster; however, the location of this gene has not been unequivocally defined . Previous characterization of the nucleotide sequence of the wec gene cluster in the region between o416 and wecG revealed that it contained three open reading frames: o74, o204, and o450 . In contrast, the results of experiments described in the current investigation revealed that it contains only two open reading frames, o359 and o450 . Mutants of E . coli possessing null mutations in o359 were unable to synthesize ECA, and they accumulated lipid II . In addition, the in vitro incorporation of {(3)H}FucNAc from TDP-{(3)H}Fuc4NAc into lipid II was not observed in reaction mixtures using cell extracts obtained from these mutants as a source of enzyme . The ECA-negative phenotype of these mutants was complemented by plasmid constructs containing the wild-type o359 allele, and Fuc4NAc transferase activity was demonstrated by using cell extracts obtained from the complemented mutants . Furthermore, partially purified o359 gene product, expressed as recombinant C-terminal His-tagged protein, was able to catalyze the in vitro transfer of {(3)H}Fuc4NAc from TDP-{(3)H}Fuc4NAc to lipid II . Our data support the conclusion that o359 of the wec gene cluster of E . coli is the structural gene for the TDP-Fuc4NAc:lipid II Fuc4NAc transferase involved in the synthesis ECA trisaccharide repeat units. Pathol Biol (Paris), 2001 Sep, 49(7), 540 - 7 {Comparison of 2 administration protocols (continuous or discontinuous) of a time-dependent antibiotic, Tazocin}; Pedeboscq S et al.; A predictive parameter of beta-lactam therapeutic efficacy is the time (T > MIC) while antibiotic serum concentrations are above the MIC of suspected bacteriological agents . This led us to carry out a randomised open study to compare the usually used intermittent administration of Tazocin (three injections of 4 g/0.5 g a day) and continuous perfusion of 12 g/1.5 g a day by calculating these T > MIC . Patients from digestive reanimation department were randomised within two arms: continuous or intermittent administration . Sixteen takings of blood were executed over a forty-hour period . After liquid/liquid extraction, piperacillin and tazobactam serum concentrations were determined by HPLC with a reversed phase column (C18) and a UV spectrophotometry detection . Then, from the time-concentration curves we have evaluated the T > MIC for an enterobacteria (MIC = 8 micrograms/mL) and for Pseudomonas (MIC = 16 micrograms/mL) . Concerning intermittent administration T > MIC were 74% (c > MICenterobacteria) and 62% (c > MICPseudomonas) . These percentages in the continuous arm were 100% (c > MICenterobacteria) and 99% (c > MICPseudomonas) . Tazobactam concentrations were low and even undetectable between each injection in the intermittent administration arm . This was not found within the continuous administration arm . In conclusion, for the intermittent administration, we observed some long periods occurring before each injection while antibiotic concentrations were under the MIC of most bacteria . During these same periods tazobactam concentrations were under the efficacy threshold . These periods were not observed within the continuous administration arm. Pathol Biol (Paris), 2001 Sep, 49(7), 515 - 21 {Spread of Enterobacteriaceae producing broad-spectrum beta-lactamase and the development of their incidence over a 16-month period in a university hospital center}; Eveillard M et al.; Enterobacteriaceae producing extended-spectrum beta-lactamases (ESBLE) constitute with methicillin-resistant Staphylococcus aureus the main multiresistant bacteria recovered in French hospitals . Our objectives were to evaluate these ESBLE diffusion in our teaching hospital and to follow their incidence during a 16-month period, whereas a control programme (barrier precautions) had been implemented in the beginning of 1999 . This study was conducted in a teaching hospital containing 1800 beds, from February 1999 to May 2000 . All ESBLE isolated in clinical or screening samples were included . Duplicates (same bacteria in the same patient) were excluded . The detection of the ESBL was performed with the double-disk diffusion test . Incidence densities were determined with their 95% confidence interval (CI95%) . Their evolution by four-month period was evaluated with the chi-square test for trend . During the 16-month period, 229 ESBLE were isolated . The incidence was 0.35 per 1000 patient-days (PD) (CI95% = {0.30-0.40}) for the whole hospital . It was 0.47/1000 PD (CI95% = {0.38-0.56}) in medical wards, 0.29/1000 PD (CI95% = {0.20-0.38}) in surgical wards and 1.32/1000 PD (CI95% = {0.90-1.74}) in intensive care units . Enterobacter aerogenes strains represented more than 75% of all ESBLE, whereas Klebsiella pneumoniae stains represented only 8.6% . During the study, the incidence of ESBLE and the proportion of strains acquired in our hospital decreased significantly (p < 0.0001 and p < 0.001 respectively) . Indeed, between the first eight-month period and the last one, the incidence of ESBLE acquired in our hospital decreased by 55%, whereas the incidence of imported strains increased slightly . This study shows that the diffusion of ESBLE concerns the entire hospital . The implementation of a control programme of the spread of multiresistant bacteria allowed us to reduce significantly the incidence of ESBLE . This incidence seemed to be stable for several months . The implementation of a policy which restricts antimicrobial use would allow us to complete the the efficacy of barrier precautions. Clin Pharmacokinet, 2001, 40(9), 685 - 94 Clinical use of ceftriaxone: a pharmacokinetic-pharmacodynamic perspective on the impact of minimum inhibitory concentration and serum protein binding; Perry TR et al.; Ceftriaxone is a third-generation cephalosporin that is used for a variety of infections such as meningitis, gonorrhoea and community-acquired pneumonia . The most important aspects of its pharmacokinetics include a long half-life, excellent tissue penetration and saturable (dose-dependent) serum protein binding of the drug . A pharmacodynamic analysis {total area under the concentration-time curve (AUC)/minimum inhibitory concentration (MIC)} was performed in several populations (healthy volunteers, children, the elderly, and patients with renal and hepatic impairment) against various bacterial species (Streptococcus pneumoniae, the Enterobacteriacieae, methicillin-susceptible Staphylococcus aureus, and Pseudomonas aeruginosa) . AUC/MIC {area under the inhibitory time curve (AUIC)} was chosen as the pharmacodynamic parameter for this analysis since ceftriaxone is a time-dependent killer and high peak concentrations are not needed . In addition, there is a significant correlation between AUIC, time when concentration exceeds the MIC (t > MIC) and time to eradication . Total and free AUICs (assuming a free fraction = 10%) were calculated since it is highly protein bound . It was postulated that a free AUIC of at least 125 would be required to achieve efficacy . From our analysis of these various populations, we were able to conclude that the free AUIC values support the use of Ig daily in infections where MIC values are below 2 mg/L . In addition, consistent with its reported good activity against CSF organisms with MICs < or =1.0 mg/L and marginal activity against organisms with MICs > or =2.0 mg/L, we also recommend the target free AUIC values of at least 125 for patients with severe infections such as meningitis . Patients with mild infections may recover with values below 125 but they may remain at risk of the development of resistant organisms . Furthermore, it is essential to further validate these findings in patients who have received treatment, calculate AUICs and correlate these parameters with both clinical and microbiological outcomes. Biofizika, 2001 Sep-Oct, 46(5), 885 - 93 {Regularities and cellular mechanism of spontaneous mutations in enterobacteria}; Chernoshchekov KA et al.; The regularities and a possible mechanism of the formation of spontaneous mutations in enterobacteria were studied . Possible causes and the mechanism of these processes were analyzed . It was shown that the mechanism of formation of spontaneous mutations is not related to the replication process and DNA polymerase errors . Mutations arise abruptly within 10-20 min due to a cosmophysical factor of unknown origin . It is assumed that cosmophysical radiation makes cell membranes excitable . Upon membrane pulsation, different volumes (portions) of cell substance are ejected through expanding pores . From this substance, viable heteromorphous mutant forms of colonies and cells (bio- and serum variants, L-forms, "parastrains", and new ecoforms of bacteria) are formed. J Am Vet Med Assoc, 2001 Oct 1, 219(7), 976 - 81 Bacteremia associated with naturally occuring acute coliform mastitis in dairy cows; Wenz JR et al.; OBJECTIVE: To determine the incidence of bacteremia in dairy cows with naturally occurring acute coliform mastitis (ACM) with a wide range of disease severity . DESIGN: Cohort study . ANIMALS: 144 dairy cows with ACM from 6 herds . PROCEDURE: Cows were examined at time of identification of ACM (time 0) and classified as having mild, moderate, or severe mastitis on the basis of rectal temperature, hydration status, rumen contraction rate, and attitude . Cows were reexamined at 24 or 48 hours . Bacteriologic culturing of milk and blood (30 ml), CBC, and serum biochemical analysis were performed at each time point . Appropriate samples were obtained at a single point from herdmates without mastitis (controls) that were closely matched for lactation number and days since parturition . Blood culture results were compared among severity groups and controls by use of chi2 tests, as was outcome of an ACM episode for cows grouped by blood bacterial isolates . RESULTS: Bacteria were isolated from 52 blood samples from 46 of 144 (32%) cows with ACM, which was significantly more than control cows (11/156; 7.1%) . Group-1 isolates (Escherichia coli, Pasteurella multocida, Mannheimia haemolytica, Klebsiella pneumoniae, Enterobacter agglomerans, and Salmonella enterica serotype Typhimurium) were identified in 20 of 144 (14%) cows with ACM and 0 of 156 control cows . Group-1 isolates were identified in 4.3, 9.1, and 42% of cows classified as having mild, moderate, and severe ACM, respectively . Escherichia coli and K pneumoniae milk and blood isolates obtained from the same cow were of the same genotype . Bacillus spp were identified in 21 of 144 (15%) cows with ACM, which was significantly more than control cows (3/156; 1.9%) . Thirty-five percent of cows with a group-1 isolate died during the mastitis episode . CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that bacteremia develops in a substantial proportion of cows with ACM . Classification of severity of disease is important for establishment of effective treatment protocols; parenteral antimicrobial treatment may be indicated in cows with ACM. Antimicrob Agents Chemother, 2001 Nov, 45(11), 3156 - 61 In vitro anti-Helicobacter pylori activity of the probiotic strain Bacillus subtilis 3 is due to secretion of antibiotics; Pinchuk IV et al.; A limited number of antibiotics can be used against Helicobacter pylori infection, and resistance jeopardizes the success of treatment . Therefore, a search for new agents is warranted . The use of probiotics to enhance gastrointestinal health has been proposed for many years, but the scientific basis of the prophylactic and therapeutic actions of probiotics has not yet been clearly delineated . Probiotic strain Bacillus subtilis 3, whose safety has previously been demonstrated, is known to have antagonistic properties against species of the family Enterobacteriaceae . In the present study, it was also found to inhibit H . pylori . The anti-H . pylori activity present in the cell-free supernatant was not related to pH or organic acid concentration . It was heat stable and protease insensitive . At least two antibiotics, detected by thin-layer chromatography (R(f) values, 0.47 and 0.85, respectively) and confirmed by high-performance liquid chromatographic analysis, were found to be responsible for this anti-H . pylori activity . All H . pylori strains tested were sensitive to both compounds . One of these compounds was identified as amicoumacin A, an antibiotic with anti-inflammatory properties . MICs for H . pylori determined in solid and liquid media ranged between 1.7 and 6.8 microg/ml and 0.75 and 2.5 microg/ml, respectively . The underestimation of MICs determined in solid medium may be due to physicochemical instability of the antibiotic under these test conditions . An additive effect between amicoumacin A and the nonamicoumacin antibiotic against H . pylori was demonstrated. J Med Microbiol, 2001 Oct, 50(10), 865 - 9 Non-standard biological activities of lipopolysaccharide from Helicobacter pylori; Matsuyama N et al.; As assessed by the lipopolysaccharide (LPS)-specific chromogenic Limulus amoebocyte lysate (LAL) assay, Helicobacter pylori LPS extracted by the phenol-water procedure showed full potency to coagulate LAL, as did LPS from Salmonella minnesota and Escherichia coli . However, pretreatment of H . pylori LPS with polymyxin B, which easily destroys the endotoxic activity of enterobacterial LPS/lipid A, had little effect on the LAL coagulation activity, although the same treatment of E . coli LPS markedly diminished its activity . The H . pylori LPS induced very weak production of nitric oxide (NO) or tumour necrosis factor (TNF) by murine macrophages and TNF by human peripheral whole blood in vitro in comparison with S . minnesota LPS . These findings indicate that H . pylori LPS has the unique endotoxic characteristic of retaining full LAL coagulation activity with polymyxin B resistance, despite losing its endotoxic potencies such as the ability to induce NO and TNF production. J Physiol Paris, 2001 Jan-Dec, 95(1-6), 51 - 7 Protection by aspirin of indomethacin-induced small intestinal damage in rats: mediation by salicylic acid; Takeuchi K et al.; Most of non-steroidal anti-inflammatory drugs (NSAIDs) except aspirin (ASA) produce intestinal damage in rats . In the present study, we re-examined the intestinal toxic effect of ASA in rats, in comparison with various NSAIDs, and investigated why ASA does not cause damage in the small intestine, in relation to its metabolite salicylic acid (SA) . Various NSAIDs (indomethacin; 10 mg/kg; flurbiprofen; 20 mg/kg; naproxen; 40 mg/kg; dicrofenac; 40 mg/kg; ASA; 20-200 mg/kg) were administered s.c., and the small intestinal mucosa was examined macroscopically 24 h later . All NSAIDs tested, except ASA, caused hemorrhagic lesions in the small intestine, with a decrease of mucosal PGE(2) contents . ASA did not provoke any damage, despite inhibiting (prostaglandin) PG production, and prevented the occurrence of intestinal lesions induced by indomethacin, in a dose-related manner . This protective action of ASA was mimicked by the equimolar doses of SA (17.8-178 mg/kg) . Indomethacin caused intestinal hypermotility, in preceding to the occurrence of lesion, and this event was followed by increases of enterobacterial translocation in the mucosa . Both ASA and SA prevented both the intestinal hypermotility and the bacterial translocation seen after indomethacin treatment . In addition, the protective effect of SA was not significantly influenced by either the adenosine deaminase or the adenosine receptor antagonists . Following administration of ASA, the blood SA levels reached a peak within 30 min and remained elevated for more than 7 h . These results suggest that SA has a cytoprotective action against indomethacin-induced small intestinal lesions, and this action may be associated with inhibition of the intestinal hypermotility and the bacterial translocation, but not mediated by endogenous adenosine . Failure of ASA to induce intestinal damage may be explained, at least partly, by a protective action of SA, the metabolite of ASA. BMC Infect Dis . 2001;1(1):16 . Epub 2001 Sep 24. The efficacy of chemical agents in cleaning and disinfection programs; Penna TC et al.; BACKGROUND: Due to the growing number of outbreaks of infection in hospital nurseries, it becomes essential to set up a sanitation program that indicates that the appropriate chemical agent was chosen for application in the most effective way . METHOD: For the purpose of evaluating the efficacy of a chemical agent, the minimum inhibitory concentration (MIC) was reached by the classic method of successive broth dilutions . The reference bacteria utilized were Bacillus subtilis var . globigii ATCC 9372, Bacillus stearothermophilus ATCC 7953, Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923 . The strains of Enterobacter cloacae IAL 1976 (Adolfo Lutz Institute), Serratia marcescens IAL 1478 and Acinetobactev calcoaceticus IAL 124 (ATCC 19606), were isolated from material collected from babies involved in outbreaks of infection in hospital nurseries . RESULTS: The MIC intervals, which reduced bacteria populations over 08 log10, were: 59 to 156 mg/L of quaternarium ammonium compounds (QACs); 63 to 10000 mg/L of chlorhexidine digluconate; 1375 to 3250 mg/L of glutaraldehyde; 39 to 246 mg/L of formaldehyde; 43750 to 87500 mg/L of isopropanol or ethanol; 1250 to 6250 mg/L of iodine in polyvinyl-pyrolidone complexes, 150 to 4491 mg/L of chlorine-releasing-agents (CRAs); 469 to 2500 mg/L of hydrogen peroxide; and, 2310 to 18500 mg/L of peracetic acid . CONCLUSIONS: Chlorhexidine showed non inhibitory activity over germinating spores . A . calcoaceticus, was observed to show resistance to the majority of the agents tested, followed by E . cloacae and S . marcescens. Clin Infect Dis, 2001 Nov 1, 33(9), 1462 - 8 Epub 2001 Oct 04. Parallel analysis of individual and aggregated data on antibiotic exposure and resistance in gram-negative bacilli; Harbarth S et al.; To evaluate the potential bias of analyzing aggregated data, we separately examined antibiotic exposure and resistance data for 35,423 patients admitted to a university hospital in Utah, from both an individual-patient perspective and group-level perspective . From 1994 through 1998, use of defined daily doses (per 1000 patient-days) of fluoroquinolones, third-generation cephalosporins, ampicillin-sulbactam, and imipenem increased by 82%, 38%, and 99%, and decreased by 38%, respectively, whereas group-level resistance rates of Enterobacteriaceae or Pseudomonas species changed only minimally . However, in individual-patient-level analyses performed by multivariable proportional hazards regression, exposure to a fluoroquinolone, third-generation cephalosporin, ampicillin-sulbactam, or imipenem was a strong risk factor for resistance to fluoroquinolones (adjusted hazard ratio {AHR}, 4.0; P<.001), third-generation cephalosporins (AHR, 3.5; P<.001), ampicillin-sulbactam (AHR, 2.3; P=.008), or imipenem (AHR, 5.7; P<.001), respectively . Thus, group-level and individual-patient-level analyses of antibiotic-use-versus-susceptibility relations yielded divergent results . Multicenter studies should include individual-patient-level data to elucidate more fully the relation between antibiotic exposure and resistance. J Spinal Cord Med, 2001 Summer, 24(2), 96 - 100 Bacteremia after spinal cord injury in initial versus subsequent hospitalizations; Waites KB et al.; BACKGROUND: Individuals with spinal cord injury (SCI) have a high lifelong risk for systemic infection . For optimal therapy, it is important to characterize the organisms involved in bacteremic episodes and the sites of primary infection . The increase in drug-resistant bacteria in recent years underscores the importance of gathering accurate microbiological information . METHODS: We performed a retrospective study of hospitalized people with SCI using a computerized Microbiology Laboratory Database . We compared the microbiology of bacteremic episodes during initial versus unplanned subsequent hospitalizations . Data were collected on 55 bacteremic episodes in 30 people during initial hospitalization for SCI and 50 episodes in 29 people who were rehospitalized . RESULTS: Among cases in which a site of origin could be identified, the respiratory tract was the origin of the majority of bacteremias during initial hospitalizations, and the urinary tract was the primary origin during rehospitalizations . Polymicrobial bacteremia occurred in 14 of 55 (25%) initial versus 14 of 50 (28%) subsequent hospitalization episodes . The most common pathogens were coagulase-negative staphylococci, followed by Staphylococcus aureus and Enterobacteriaceae . Bacteremia was more common in people with tetraplegia and complete neurologic lesions than in those with paraplegia and incomplete lesions . One person in the rehospitalization group died from complications of bacteremia . All others were successfully treated . CONCLUSIONS: This study describes the frequency and characteristics of bacteremia during initial and subsequent hospitalizations following SCI and examines differences in original sites of infection . This information should be considered when planning infection control measures and empiric antibiotic regimens for patients with SCI. Clin Microbiol Rev, 2001 Oct, 14(4), 933 - 51, table of contents Extended-spectrum beta-lactamases in the 21st century: characterization, epidemiology, and detection of this important resistance threat; Bradford PA; Beta-lactamases continue to be the leading cause of resistance to beta-lactam antibiotics among gram-negative bacteria . In recent years there has been an increased incidence and prevalence of extended-spectrum beta-lactamases (ESBLs), enzymes that hydrolyze and cause resistance to oxyimino-cephalosporins and aztreonam . The majority of ESBLs are derived from the widespread broad-spectrum beta-lactamases TEM-1 and SHV-1 . There are also new families of ESBLs, including the CTX-M and OXA-type enzymes as well as novel, unrelated beta-lactamases . Several different methods for the detection of ESBLs in clinical isolates have been suggested . While each of the tests has merit, none of the tests is able to detect all of the ESBLs encountered . ESBLs have become widespread throughout the world and are now found in a significant percentage of Escherichia coli and Klebsiella pneumoniae strains in certain countries . They have also been found in other Enterobacteriaceae strains and Pseudomonas aeruginosa . Strains expressing these beta-lactamases will present a host of therapeutic challenges as we head into the 21st century. Arch Pediatr, 2001 Sep, 8 Suppl 4, 721s - 725s {Severe neonatal bacterial infections}; Saizou C et al.; The prognosis of septicemic forms of early and late neonatal sepsis is severe with a high rate of mortality especially in premature infants . The evaluation of severity is difficult because of the non specificity of the clinical signs and mortality seems to be a good means of evaluation . A study was conducted in France on the mortality due to infection in neonatal intensive care units and neonatology wards during the third trimester 2000 . Among 18 units, the mortality rate was 9.3% of admissions, corresponding to 11 early onset sepsis and 17 nosocomial infections . Death in primitive infections is essentially due to group B streptococci and E . coli with a more important risk in low gestational age infants . The nosocomial infections arise almost only in premature infant . Prognosis of infections due to Staphylococcus coagulase negative staphylococci, most frequent pathogens is good but mortalities rate is higher for enterobacteriacae--40% and for Pseudomonas, 62%. Arch Pediatr, 2001 Sep, 8 Suppl 4, 697s - 704s {Impact of bacterial resistance on severe infections}; Raymond J et al.; Since many years, the antimicrobial resistance increases as well as for community-acquired as for nosocomial infections . Antibiotic-resistant pneumococci are neither more nor less virulent susceptible strains . Except for immunocompromised patients, the outcome of penicillin-resistant pneumococcal infections have been similar to those in patients who are infected by susceptible ones . Current levels of S . pneumoniae resistance to penicillin and cephalosporin are not associated to an increase in mortality in children with meningitis if adequate doses of antibiotics are given . Because empiric therapy has changed, antibiotic resistance has not been associated with increased mortality . This statement can be extended to Meningococcus, for which 32 to 50% of the strains have a decreased susceptibility to penicillin . For nosocomial infections, S . aureus is the main studied pathogen . Several studies report that in patients with severe diseases (bacteremia or pneumonia) methicillin resistance of S . aureus had no significant impact on patient outcome after adjustment for different confounders . The main risk factor for mortality is the severe underlying diseases rather than the resistance as well for methicillin--resistant S . aureus, as for vancomycin resistant enterococci, Klebsiella with extended spectrum beta lactamase and Enterobacters . Recommendations for controlling epidemiologic surveillance, using barrier precautions and limiting the use of antibiotics as well in the hospital as in the community must be undertaken. Ophthalmology, 2001 Oct, 108(10), 1830 - 4 Bacillus cereus keratitis associated with contact lens wear; Pinna A et al.; OBJECTIVE: We report the first case of contact lens-related Bacillus cereus keratitis and ulcer associated with B . cereus contamination of the contact lens case . This is also the first study to investigate and establish the genetic identity of an organism isolated from the cornea and contact lens case in a patient with contact lens-associated keratitis . DESIGN: Case report . INTERVENTION AND TESTING: Conjunctival swabs and corneal scrapings from the left eye were inoculated for culture . The contact lens case was also cultured . Antibiotic susceptibility testing was determined by agar disk diffusion method . Initial treatment with topical ciprofloxacin and fortified tobramycin was given . Genetic analysis of the bacterial isolates was performed using polymerase chain reaction (PCR) with enterobacterial repetitive intergenic consensus primers (ERIC; ERIC-PCR) . Susceptibility of B . cereus to heat and contact lens disinfecting solutions containing hydrogen peroxide, hydrogen peroxide-catalase, polyquaternium-1, and polyaminopropyl biguanide (PAPB) was tested . MAIN OUTCOME MEASURES: Clinical features, culture results, and antibiotic susceptibility testing were analyzed . The ERIC-PCR amplification products were visualized in ethidium bromide-stained agarose gel . Bacterial growth after exposure to heat and contact lens disinfecting solutions was assessed on blood agar plates . RESULTS: B . cereus was grown from the conjunctiva, corneal ulcer, and contact lens case . All isolates were sensitive to gentamicin, tobramycin, ciprofloxacin, clindamycin, and vancomycin . The corneal ulcer gradually healed over the next 6 days . Results of ERIC-PCR showed that the isolates from the cornea and contact lens case were indistinguishable, thus demonstrating the source of infecting organism to be the contaminated contact lens case . Exposure to a temperature of 80 degrees C for 20 minutes and incubation with hydrogen peroxide-catalase, polyquaternium-1, and PAPB for the minimum recommended time failed to kill B . cereus . Only exposure to hydrogen peroxide for 4 hours eradicated the organism . CONCLUSIONS: B . cereus should be considered a possible etiologic agent of contact lens-associated keratitis . Heat and many types of contact lens disinfecting solutions may be ineffective in eradicating B . cereus from contaminated contact lens cases . Only prolonged exposure to hydrogen peroxide appeared to be sporicidal to B . cereus in this study. Diagn Microbiol Infect Dis, 2001 Aug, 40(4), 199 - 201 Enzymatic characterization of TEM-63, a TEM-type extended spectrum beta-lactamase expressed in three different genera of Enterobacteriaceae from South Africa; Hanson ND et al.; The extended-spectrum beta-lactamase, TEM-63, was identified in three separate genera of South African isolates: Proteus mirabilis, Klebsiella pneumoniae, and Escherichia coli . This paper describes identification of the gene in these isolates and compares relative rates of hyrolysis between TEM-63 and other known ceftazidimases. Diagn Microbiol Infect Dis, 2001 Aug, 40(4), 179 - 86 Evaluation of E test, disk diffusion and broth microdilution to establish tentative quality control limits and review susceptibility breakpoints for two aerobic actinomycetes; Tomlin P et al.; A single laboratory study was carried out to compare E Test with broth microdilution and disk diffusion to establish tentative quality control ranges for Nocardia asteroides ATCC 19247 and Rhodococcus equi ATCC 6939 against a panel of eight antimicrobial agents . Reproducibility testing was performed on 12 consecutive days to establish tentative quality control ranges . A total of 36 clinical strains of the Nocardia asteroides complex and 5 Rhodococcus strains were used in the study . Both candidate control strains and clinical strains grew well on cation-adjusted Mueller-Hinton agar . Adequate growth occurred at 48 to 72 h for the Nocardia isolates and 24 to 48 h for Rhodococcus . A standardized primary inoculum of 5 x 10(4) CFU/mL was used for performance of E Test and disk diffusion for the Nocardia isolates . Tentative population-based error rates were calculated using current breakpoints for Enterobacteriaceae for E Test compared with disk diffusion for the 36 clinical strains of Nocardia species . Significant very major error rates were observed for imipenem (22%) and minor error rates varied from 2.7% to 50% . These methods require more extensive validation before definitive breakpoint criteria can be established. Diagn Microbiol Infect Dis, 2001 Aug, 40(4), 173 - 7 In vitro antimicrobial activity of GAR-936 tested against antibiotic-resistant gram-positive blood stream infection isolates and strains producing extended-spectrum beta-lactamases; Biedenbach DJ et al.; GAR-936, a new, semisynthetic glycylcycline, has shown good antibacterial activity against a wide range of clinically important Gram-positive and -negative aerobic bacteria including Streptococcus pneumoniae, Hemophilus influenzae, Moraxella catarrhalis, Neisseria gonorrhoeae, most Enterobacteriaceae, Staphylococcus aureus and Enterococcus spp . The purpose of this study was to determine the activity of GAR-936 against a range of Gram-positive and -negative bloodstream isolates including many strains producing extended-spectrum beta-lactamases (ESBLs) . Six hundred four bloodstream isolates of Gram-positive cocci collected as part of the SENTRY surveillance program were selected for their geographic diversity . GAR-936 was also tested against an additional 176 Gram-negative cocci isolates (Klebsiella pneumoniae, 98 strains; Escherichia coli, 78 strains), 96 of which were ESBL-producers . Broth microdilution testing was used to determine the susceptibility of the selected organisms and a range of comparator antimicrobial agents whose choice was based on their activities against the selected pathogens and included a mix of both newer and older agents . Presence of an ESBL-producing strain was confirmed using the clavulanate test . GAR-936 demonstrated impressive activity against all 604 strains of Gram-positive cocci, with an MIC range of <or=0.015-1 microg/mL, and MIC(90)s ranging from <or=0.05 microg/mL for S . pneumoniae to 0.25 for S . aureus and coagulase-negative staphylococci . MICs against the 176 Gram-negative isolates were higher (range 0.06-4 microg/mL), with MIC(90)s of 0.25-1 microg/mL . The activity of GAR-936 was relatively unaffected by the presence of ESBLs . The activity of GAR-936 was particularly impressive against Gram-positive cocci when compared against the test results for vancomycin and the newer antimicrobial agents, linezolid and quinupristin/dalfopristin . The MIC(90)s for GAR-936 were significantly lower than the comparator agents for all species tested with particularly impressive results against S . pneumoniae and enterococci. J Microbiol Methods, 2001 Nov, 47(2), 243 - 7 Performance of Pseudomonas CFC-selective medium in the fish storage ecosystems; Tryfinopoulou P et al.; Pseudomonas agar base supplemented with cephaloridine, fucidin, and cetrimide (CFC) was used to count Pseudomonas populations on fish . Both Enterobacteriaceae and Shewanella putrefaciens were able to grow on the CFC medium . Evaluation of the performance of CFC-selective for pseudomonads medium, on fish samples stored aerobically and under a modified atmosphere at 0, 10 and 20 degrees C was tested.The selectivity of the medium was affected by storage temperatures and the type of packaging of the fish samples . The selectivity of the medium diminished as the population increased and for samples stored at high temperature (20 degrees C) or under modified atmospheres.When designing adequate selectivity of a medium, interfering organisms should be taken into account, especially when the background flora tends to be more robust than the organisms to be counted or detected. J Microbiol Methods, 2001 Nov, 47(2), 209 - 17 Rapid duplex PCR assay for the detection of pathogenic Yersinia enterocolitica strains; Aarts HJ et al.; For the detection of pathogenic Yersinia enterocolitica strains, a duplex PCR has been developed based on differences observed between the fingerprint profiles of pathogenic and non-pathogenic strains . The profiles were obtained by using a primer derived from the Enterobacterial Repetitive Intergenic Consensus (ERIC) sequences . From the sequence of one pathogen-specific amplified fragment, a discriminative primer has been designed bridging the sequence of the highly conserved core region and 3' end of the ERIC element . In combination with three other primers, all located within the detected open reading frame that resembled the sequence of the bipA gene, this primer was applied in a duplex PCR assay to simultaneously detect Y . enterocolitica and to discriminate between pathogenic and non-pathogenic strains . The same primer combinations were used in an on line rapid cycling real-time PCR assay . The used SYBR Green I format allowed for the easy translation of the PCR conditions and confirmation of the resulting amplicons . The time of analysis was reduced to approximately 60 min. Can J Microbiol, 2001 Aug, 47(8), 698 - 705 Involvement of gacS and rpoS in enhancement of the plant growth-promoting capabilities of Enterobacter cloacae CAL2 and UW4; Saleh SS et al.; The plant growth-promoting bacteria Enterobacter cloacae CAL2 and UW4 were genetically transformed with a multicopy plasmid containing an rpoS or gacS gene from Pseudomonas fluorescens . The transformed strains were compared with the nontransformed strains for growth, indoleacetic acid (IAA) production, antibiotic production, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, siderophore production, cell morphology, and the ability to promote canola root elongation . All transformed strains had a longer lag phase, were slower in reaching stationary phase, and attained a higher cell density than the nontransformed strains . Transformation resulted in cells that were significantly shorter than the nontransformed cells . The transformed strains also produced significantly more IAA than the nontransformed strains . Introduction of rpoS or gacS from Pseudomonas fluorescens was associated with a reduction in the production of both antibiotics, 2,4-diacetylphloroglucinol and mono-acetylphloroglucinol, produced by Enterobacter cloacae CAL2 . With Enterobacter cloacae CAL2, plasmid-borne rpoS, but not gacS, increased the level of ACC deaminase activity, while introduction of rpoS in Enterobacter cloacae UW4 caused a decrease in ACC deaminase activity . Neither gacS nor rpoS significantly affected the level of siderophores synthesized by either bacterial strain . Overproduction of either GacA or RpoS in Enterobacter cloacae CAL2 resulted in a significant increase in the root lengths of canola seedlings when seeds were treated with the bacteria, and overproduction of RpoS caused an increase in canola shoot as well as root lengths. Blood Purif, 2001, 19(4), 380 - 7 Blood components influence cytokine induction by bacterial substances; Schindler R et al.; While some studies clearly demonstrated transfer of cytokine-inducing substances (CIS) through dialysis membranes, other authors were unable to reproduce such a transfer . This inconsistency may have been caused by marked differences in experimental design . We performed a systematic evaluation of cytokine induction in whole blood and from separated peripheral blood mononuclear cells (PBMC) using either purified lipopolysaccharide (LPS) or culture supernatants from various bacterial strains . An in vitro hemodialysis circuit with whole blood in the blood compartment was employed; the dialysate was contaminated with sterile filtrates from Pseudomonas aeruginosa cultures . Addition of plasma samples from the blood side to PBMC readily induced interleukin (IL)-1beta and IL-6 after contamination of the dialysate, while the same samples failed to induce cytokine production in whole blood . In experiments using direct incubation, purified P . aeruginosa LPS induced more IL-1beta and IL-6 in whole blood compared to PBMC . In contrast, bacterial filtrates from Escherichia coli, Enterobacter cloacae and especially P . aeruginosa induced significantly less cytokines in whole blood compared to PBMC . Addition of erythrocytes but not granulocytes decreased cytokine induction by bacterial filtrates as well as by LPS, probably by adsorption of these substances . The addition of 30% plasma increased cytokine induction by LPS but decreased cytokine induction by P . aeruginosas filtrates . Our results suggest that cellular and plasmatic components of whole blood interact with bacterial CIS altering their pyrogenic activity . The detection of CIS depends on the test system used; whole blood cultures are not as sensitive for the detection of CIS from bacterial filtrates as PBMC cultures . J Clin Microbiol, 2001 Oct, 39(10), 3772 - 4 Contamination of the clinical microbiology laboratory with vancomycin-resistant enterococci and multidrug- resistant Enterobacteriaceae: implications for hospital and laboratory workers; Collins SM et al.; We surveyed environmental surfaces in our clinical microbiology laboratory to determine the prevalence of vancomycin-resistant enterococci (VRE) and multidrug-resistant Enterobacteriaceae (MDRE) during a routine working day . From a total of 193 surfaces, VRE were present on 20 (10%) and MDRE were present on 4 (2%) of the surfaces tested . In a subsequent survey after routine cleaning, all of the 24 prior positive surfaces were found to be negative . Thus, those in the laboratory should recognize that many surfaces may be contaminated by resistant organisms during routine processing of patient specimens. J Clin Microbiol, 2001 Oct, 39(10), 3724 - 6 Increased prevalence of class I integrons in Escherichia coli, Klebsiella species, and Enterobacter species isolates over a 7-year period in a German university hospital; Schmitz FJ et al.; The prevalence of integrons in five enterobacterial species was analyzed in 900 blood culture isolates from 1993, 1996, and 1999 . Remarkably, the prevalence increased from 4.7% in 1993 to 9.7% in 1996 and finally to 17.4% in 1999 (P < 0.01) . Within 7 years the combined percentage of P1 strong promoters and P1 weak plus P2 active promoters with high transcription efficacies has increased from 23.1 to 33.3 and finally 60% (P < 0.05). Biochem Soc Symp, 2001, (68), 143 - 53 Degradation of explosives by nitrate ester reductases; Williams RE et al.; Explosive-contaminated land poses a hazard both to the environment and to human health . Microbial enzymes, either in their native or heterologous hosts, are a powerful and low-cost tool for eliminating this environmental hazard . As many explosives have only been present in the environment for 10 years, and with similar molecules not known in Nature, the origin of enzymes specialized for the breakdown of explosives is of particular interest . Screening of environmental isolates resulted in the discovery of flavoproteins capable of denitrating the explosives pentaerythritol tetranitrate (PETN) and glycerol trinitrate . These nitrate ester reductases are related in sequence and structure to Old Yellow Enzyme from Saccharomyces carlsbergenisis . All the members of this family have alpha/beta barrel structures and FMN as a prosthetic group, and reduce various electrophilic substrates . The nitrate ester reductases are, however, unusual in that they display activity towards the highly recalcitrant, aromatic explosive 2,4,6-trinitrotoluene, via a reductive pathway resulting in nitrogen liberation . We have embarked on a detailed study of the structure and mechanism of PETN reductase from a strain of Enterobacter cloacae . Work is focused currently on relating structure and function within this growing family of enzymes, with a view to engineering novel enzymes exhibiting useful characteristics. Antibiot Khimioter, 2001, 46(6), 12 - 20 {Monitoring of uropathogens and their susceptibility to antibiotics}; Savitskaia KI et al.; Samples of urine collected from patients with complicated urology infection and hospitalized to the Moscow Region Research Clinical Institute in 1986, 1991, 1995 and 1999 were analysed . Of 11,444 samples examined, bacteriuria was estimated in 7143 samples . 9786 strains (29 genus) of bacteria were isolated--56.9 per cent as mono culture and 43.1 per cent as associations . Susceptibility to 21 antibiotic was determined by disk diffusion method for 1607 strains; beta-lactamase production was determined in 198 strains, MIC was determined for 41 antibiotics . Gram-negative rods relative amount among pathogens decreased substantially (84.7 per cent in 1986 against 61.6 per cent in 1999), particularly Enterobacteriaceae (74.7 per cent in 1986 against 41.4 per cent in 1999) . Nonfermenting Gram-negative rods (NFGNR) relative amount increased (10.8 per cent against 19.2 per cent), along with Gram-positive cocci (19.8 per cent against 64.2 per cent), particularly coagulasenegative staphylococci (CNS) (10.8 per cent against 35.9 per cent) and enterococci (5 per cent against 16.5 per cent) and candida and fungi (0.5 per cent in 1986 against 15.9 per cent in 1999) . At the period 1986-1999 the main pathogens in urology infection were E . coli, Enterobacter spp., NFGNR (including P . aeruginosa), Staphylococcus, CNS, Enterococcus spp . The problem pathogens for urological department were the following: E . coli, Klebsiella spp., Enterobacter spp., Proteus spp., NFGNR including P . aeruginosa, CNS, Enterococcus spp., candida and fungi . At the period 1991-1997 Gram-negative pathogens susceptibility to amikacin, ofloxacin, ciprofloxacin, imipenem, ceftazidime, cefotaxime was not changed in general, Gram-positive cocci (staphylococci and enterococci) retained the same susceptibility to vancomicin, cefamandol and amoxyclave . Staphylococci were also susceptible to amikacin, imipenem, rifampicin, oxacillin, ciprofloxacin, and ofloxacin . Production of beta-lactamase was registered for 38.7 per cent of CNS, 26.5 per cent of E . coli, 38.5 per cent of K . pneumoniae, 25 per cent of P . mirabilis and 55.6 per cent of P . aeruginosa strains. Appl Environ Microbiol, 2001 Oct, 67(10), 4760 - 4 Survival of salmonellae on and in tomato plants from the time of inoculation at flowering and early stages of fruit development through fruit ripening; Guo X et al.; The fate of salmonellae applied to tomato plants was investigated . Five Salmonella serotypes were used to inoculate tomato plants before and after fruits set, either by injecting stems with inoculum or brushing flowers with it . Ripe tomato fruits were subjected to microbiological analysis . Peptone wash water, homogenates of stem scar tissues, and homogenates of fruit pulp were serially diluted and plated on bismuth sulfite agar before and after enrichment . Presumptive Salmonella colonies were confirmed by serological tests, PCR assay using HILA2 primers, and enterobacterial repetitive intergenic consensus PCR . Of 30 tomatoes harvested from inoculated plants, 11 (37%) were positive for Salmonella . Of the Salmonella-positive tomatoes, 43 and 40%, respectively, were from plants receiving stem inoculation before and after flower set . Two of eight tomatoes produced from inoculated flowers contained Salmonella . Higher percentages of surface (82%) and stem scar tissue (73%) samples, compared to pulp of Salmonella-positive tomatoes (55%), harbored the pathogen . Of the five serotypes in the inoculum, Montevideo was the most persistent, being isolated from tomatoes 49 days after inoculation, and Poona was the most dominant, being present in 5 of 11 Salmonella-positive tomatoes . Results suggest that Salmonella cells survive in or on tomato fruits from the time of inoculation at flowering through fruit ripening . Tomato stems and flowers are possible sites at which Salmonella may attach and remain viable during fruit development, thus serving as routes or reservoirs for contaminating ripened fruit. Anal Chem, 2001 Sep 1, 73(17), 4241 - 8 Electrochemical biosensor array for the identification of microorganisms based on lectin-lipopolysaccharide recognition; Ertl P et al.; Rapid identification of bacterial strains remains a well-known problem in applied medicine and, for viable pathogens, is an important diagnostic goal . We have investigated an electrochemical biosensor array, in which transduction is based on respiratory cycle activity measurements, where the microorganism's native respiratory chain is interrupted with non-native external oxidants . The selective biochemical recognition agents employed in this study are lectins that, once immobilized, recognize and bind to cell surface lipopolysaccharides . Porous membranes with different surface properties were examined as potential immobilization supports for these lectins . Optimizations performed using concanavalin A and E . coli JM105 show that immobilization methods involving pre-activated membranes significantly reduce the time required to create a functional lectin layer on the membrane surface . Overall, we found general agreement between agglutination test results and the electrochemical assessment of lectin-cell binding . Chronocoulometric measurements were made for cells captured on lectin-modified Immunodyne ABC membranes physically affixed to Pt working electrodes . This lectin-based sensor array was exposed to viable cells of gram-negative and gram-positive bacteria as well as yeast, and chronocoulometric measurements were used to generate a pattern of responses for each organism toward each lectin . Principal component analysis was used to classify the chronocoulometric results for the different microbial strains . With this new method, six microbial species (Baccilus cereus, Staphylococcus aureus, Proteus vulgaris, Escherichia coli, Enterobacter aerogenes, Saccharomyces cerevisiae) were readily distinguished. Zh Mikrobiol Epidemiol Immunobiol, 2001 Jul-Aug, (4), 86 - 9 {Characteristics of changes in microbiocenosis in patients with chronic nonspecific urethritis}; Bukharin OV et al.; The species composition and the complex of biological characteristics of microflora in the front section of urethra in healthy males and in patients with chronic nonspecific urethritis . The study revealed that in patients with chronic nonspecific nongonococcal urethritis changes in the microbiocenosis of the urethra were observed . These changes were manifested by a decrease in the number of species, the appearance of Grain-negative enterobacterial flora and an increase in the persistence potential of symbiotic bacteria . These disturbances are regarded as the manifestation of urogenital dysbiosis. Zh Mikrobiol Epidemiol Immunobiol, 2001 Jul-Aug, (4), 107 - 11 {Characteristics of enterobacteria isolated from patients with urogenital pathology}; Gritsenko VA et al.; Species composition and a number of persistence characteristics enterobacteria isolated from urine of 42 pregnant and 22 nonpregnant women with pyelonephritis (relapse, remission), from prostatic fluid of 225 males and secretions of cervical canal of 124 women with urogenital pathology (prostatitis, salpingo-oophoritis) were studied . The study revealed that enterobacteria, including Escherichia coli, prevailed in the structure of uromicroflora (66.7-83.3%) and constituted a relatively small proportion among "genital" isolates of microorganisms (19.9-22.2%) . Male and female sterility and the presence of enterobacteria in the reproductive tract of patients were found to be directly correlated . Clinical isolates of enterobacteria were shown to possess pronounced seroresistance and the complex of persistence characteristics, including antilysozyme, anti-intercidal and anticomplementary activity. Clin Infect Dis, 2001 Nov 1, 33(9), 1513 - 9 Epub 2001 Sep 24. The microbiology of postoperative peritonitis; Roehrborn A et al.; Postoperative peritonitis carries a higher risk of complications and mortality than does community-acquired disease . Little, however, is known about the specific microbiology of this condition . To gain insight into this problem, the microbiological findings of 67 patients with postoperative peritonitis were compared with those of 68 patients with community-acquired peritonitis . In a comparison of postoperative peritonitis with community-acquired disease, the number of isolates of enterococci (23 versus 6) and Enterobacter species (13 versus 4) were increased and the number of isolates of Escherichia coli (21 versus 42) were reduced . Antibiotic therapy before reintervention increased the number of resistant organisms at relaparotomy (33% versus 8%) . The in vitro efficacy of the primary antibiotic or combination of drugs did not affect mortality rates (40% versus 38% after effective and ineffective treatment, respectively) . Thus, the microbiology of postoperative peritonitis differs significantly from that of community-acquired disease, and specific antibiotic therapy is required, despite the doubtful impact on survival. Biofactors, 2001, 14(1-4), 241 - 54 Bioremediation of selenium-contaminated sediments and water; Frankenberger WT Jr et al.; Selenium (Se) is a contaminant of agricultural irrigation-drainage water in the western United States, and the cause of wildlife deaths and grotesque deformities . Some approaches in reducing the toxic Se concentrations from contaminated sediments and water have been proposed, but most of these tend to be costly or ineffective . Bioremediation through microbial transformations of toxic Se species into nontoxic forms is being considered as an effective remedial alternative . The microbial reduction of toxic oxyanions of Se (SeO(4)(2-) and SeO(3)(2-)) into insoluble Se(0) or methylation of these species to dimethylselenide (DMSe) has been accepted as a potential bioremediation strategy for cleanup of Se-contaminated water and sediments . By conducting a series of laboratory, bench-scale and field studies, we have thoroughly investigated the remedial potential of these approaches . It was observed that microorganisms, particularly Enterobacter cloacea, are very active in reduction of Se oxyanions present in irrigation drainage water, into insoluble Se(0) and, by monitoring various environmental conditions and addition of organic amendments, the process could be stimulated manifold . Similarly, the process of biomethylation of Se in soil sediments and water was found active and highly dependent on specific carbon amendments (pectin and proteins), pH, temperature, moisture, aeration and activators (cofactors) . Moreover, Se biomethylation was protein/peptide-limited rather than nitrogen-, amino acid- or carbon-limited . Crude casein and its components were equally stimulatory producing a >50-fold enhancement in DMSe yield . Methionine and methyl cobalamin stimulated DMSe production by Alternaria alternata, indicating that the coenzyme may mediate the transfer of a methyl group to the Se atom . An acute toxicity test involving inhalation of DMSe by rats revealed that DMSe is nontoxic . Experiments were scaled up from laboratory studies to field plots to verify the feasibility of this bioremediation approach . Based upon the promising results of these studies, a biotechnology prototype was developed which could be applicable for cleanup of polluted sediments and water throughout the western United States. J Photochem Photobiol B, 2001 Sep 15, 62(3), 158 - 65 Sublethal effects of ultraviolet A radiation on Enterobacter cloacae; Oppezzo OJ et al.; We report the sublethal effects of ultraviolet A (UVA) on Enterobacter cloacae in comparison with those produced in Escherichia coli . UVA-induced sublethal effects were investigated in either bacterial membrane and at tRNA level . Limited dependence on oxygen concentration for photoinduced inhibition of biochemical membrane functions and low levels of oxidative damage during the irradiation period were found in En . cloacae . On the other hand, ultraviolet spectroscopy and reversed-phase HPLC analysis of hydrolysed tRNA showed that radio induced damage to tRNA is similar in En . cloacae and E . coli . Nevertheless, growth delay induced by UVA in En . cloacae was shorter than that found in E . coli submitted to the same experimental conditions . A limited post-irradiation ppGpp accumulation and the absence of any influence of the membrane damage on the growth delay extent seem to be responsible for the shortness of this effect in En . cloacae . Most of the differences between En . cloacae and E . coli could be attributed to an increased ability of En . cloacae to overcome oxidative stress during UVA exposure. Ross Gastroenterol Zh, 2001, (1), 54 - 69 {Comparative study of chromatography-mass spectrography of microorganism's chemical markers in blood and intestinal mucosa bioptats}; Osipov GA et al.; Fatty acids, that are microorganism's markers in blood and jejunum's, ileum's, and large intestine's bioptats of the patients with irritable bowel syndrome were investigated with gas chromatography . Markers of Cl . perfringens, Cl . difficile, Enterococcus, Streptomyces, Enterobacterial, Klebsiella, E . coli, Peptostreptococcus, Candida Albicans, genus of Streptococcus, of Staphylococcus, of Fusobacterium sp and others microorganisms were revealed . Settling of intestinal mucosa was the same in jejunum, ileum and large intestine (0.5-1.3) x 10(10) cells/gr . and was approximately the same as microorganism's concentration in feces (1.8 x 10(11) cells/gr) . Correlation of blood and bioptats markers was demonstrated in the most patients . The possibility to use specific fatty blood acids as microorganisms markers in preliminary diagnosis of mucosa microflora changes is discussed. J Clin Virol, 2001 Oct, 22(3), 217 - 27 Lipid rafts and HIV-1: from viral entry to assembly of progeny virions; Campbell SM et al.; BACKGROUND: Lipid rafts are currently an intensely investigated topic of cell biology . In addition to a demonstrated role in signal transduction of the host cell, lipid rafts serve as entry and exit sites for microbial pathogens and toxins, such as FimH-expressing enterobacteria, influenza virus, measles virus and cholera toxin . Furthermore, caveolae, a specialised form of lipid raft, are required for the conversion of the non-pathogenic prion protein to the pathogenic scrapie isoform . OBJECTIVES: A number of reports have shown, directly or indirectly, that lipid rafts are important at various stages of the human immunodeficiency virus type-1 (HIV-1) replication cycle . The purpose of this paper is to provide a brief overview of the role of membrane-associated lipid rafts in cell biology, and to evaluate how HIV-1 has hijacked this cellular component to support HIV-1 replication . Special sections are devoted to discussing the role of lipid rafts in (1) the entry of HIV-1, (2) signal transduction regulation in HIV-1-infected cells, (3) the trafficking of HIV-1 proteins via lipid rafts during HIV-1 assembly; and a further section discusses the role of cholesterol in mature HIV-1 . SUMMARY: Like a number of other pathogens, HIV-1 has evolved to rely on the host cell lipid rafts to support its propagation during multiple stages of the HIV-1 replication cycle . This review has highlighted the importance of lipid rafts in HIV-1 replication. Semin Respir Infect, 2001 Sep, 16(3), 215 - 24 Future of the quinolones; Ball P; New fluoroquinolones and fluoronaphthyridones continue to provide the mainstay of antibiotic development, despite recent events associated with unexpected or uncharacteristically severe adverse drug reactions . These have included hepatotoxicity caused by trovafloxacin (suspended), cardiotoxicity associated with grepafloxacin, and phototoxicity caused by clinafloxacin (both withdrawn) . Prolongation of the QT interval appears to be an emergent class effect, the implications of which are not yet fully understood . However, the second-generation agents ciprofloxacin and, latterly, levofloxacin have excellent safety profiles and provide standard optimal choices for therapy of a wide range of gram-negative pathogens . They are also useful for many respiratory infections, though the use of ciprofloxacin in pneumococcal pneumonia has been questioned and continued use of levofloxacin may act as a selection pressure for emergence of quinolone-resistant Streptococcus pneumoniae . Active conservation measures may be required to protect the class from this problem because alternatives, should high-level penicillin-resistance continue to spread, are few . The new 8-methoxy quinolones (moxifloxacin and gatifloxacin) are more highly potent against both penicillin-susceptible and multidrug-resistant S . pneumoniae, while retaining activity against enterobacteria . Clinical Phase III development has shown them to produce very satisfactory clinical and bacteriologic responses in respiratory infections and to be remarkably free of clinically significant adverse effects . Postmarketing surveillance of moxifloxacin in Germany has revealed no additional concerns . These agents are now licensed in many countries, including the United States, and add a further, broad-based respiratory dimension to the future of the class . Eur J Clin Microbiol Infect Dis, 2001 Jul, 20(7), 486 - 9 Ciprofloxacin- and methicillin-resistant staphylococcus aureus susceptible to moxifloxacin, levofloxacin, teicoplanin, vancomycin and linezolid; Presterl E et al.; In order to determine the comparative efficacy of vancomycin,