Saudi Med J, 2004 May, 25(5), 570 - 4 Prevalence of extended spectrum beta-lactamase among multidrug resistant gram-negative isolates from a general hospital in Saudi Arabia; Kader AA et al.; OBJECTIVE: To determine the prevalence of extended spectrum beta-lactamase (ESBL) among multidrug resistant isolates of enterobacteriaceae and non-fermenting gram-negative bacilli . METHODS: This study was carried out at the Almana General Hospital, Eastern Province, Kingdom of Saudi Arabia, during the period March 2002 through to June 2003 . Multidrug resistant gram-negative isolates from patients admitted to the surgical, medical, pediatric, long-term care and intensive care units were studied for the presence of the ESBL enzyme . RESULTS: A total of 3231 gram-negative organisms were studied for the presence of multidrug resistance and ESBLs . Of these, 197 (6%) isolates were multidrug resistant (MDR), and 156 (4.8%) were positive for ESBL . Seventy nine percent of the MDR strains were positive for ESBL . The most frequent isolates were Escherichia coli (1116) and Klebsiella pneumoniae (687) and ESBL was detected in 72 (6.5%) and 37 (5.4%) of these isolates . The MDR strains that produced ESBL were most commonly isolated from surgical care patients with diabetic fascitis (83%) and patients with indwelling Foley's catheter (79%) . Extended spectrum beta-lactamase producing strains showed the highest susceptibility to imipenem and meropenem (86%) . The non-beta-lactam antibiotics with greatest activity against these ESBL strains in vitro were ciprofloxacin (72%), amikacin (70%), tobramycin (67%) and gentamicin (56%) . CONCLUSION: The majority (79%) of the MDR enterobacteriaceae and non-fermenting gram-negative bacilli tested over 15-months were positive for ESBL . Imipenem, meropenem, ciprofloxacin and amikacin showed the highest activity against these ESBL-producing organisms . Due to the growing problem of infection with ESBL-producing bacteria, which are frequently resistant to many classes of antibiotics resulting in difficult-to-treat infections, clinicians need to be familiar with the clinical significance of these enzymes and potential strategies for dealing with them. FEMS Microbiol Lett, 2004 May 15, 234(2), 209 - 13 Novel GES/IBC extended-spectrum beta-lactamase variants with carbapenemase activity in clinical enterobacteria; Vourli S et al.; Two clinical isolates, an Escherichia coli and a Klebsiella pneumoniae, with decreased susceptibility to carbapenems were studied . This phenotype was associated with production of novel GES/IBC variant beta-lactamases, designated GES-3 (from E . coli) and GES-4 (from K . pneumoniae), exhibiting carbapenemase activity . Both enzymes possessed Ser at Ambler's position 170 instead of Gly found in the beta-lactamases GES-1 and IBC-1 that lack carbapenemase activity . Additionally, position 104 in GES-4 was occupied by a Lys as in IBC-1 . bla(GES-3) and bla(GES-4) occurred as gene cassettes in the variable regions of class 1 integrons carried by plasmids . The structure of the GES-4-encoding integron was similar to that of previously described IBC-1 integrons . The GES-3-encoding integron was, most likely, truncated at the 3' conserved segment . FEMS Microbiol Lett, 2004 May 15, 234(2), 201 - 7 Diversity of genetic environment of bla(CTX-M) genes; Lartigue MF et al.; Twenty-four non-clonally related enterobacterial isolates producing the emerging CTX-M-type extended-spectrum beta-lactamases were recovered from several countries including France, India, Poland, and Turkey . They had been isolated from 2000 to 2003 . beta-Lactamases CTX-M-2, CTX-M-3, CTX-M-10, CTX-M-14 and CTX-M-15 were identified . Most of the isolates produced beta-lactamase CTX-M-15 . Insertion sequence ISEcp1 was found upstream of bla(CTX-M-3), bla(CTX-M-10), bla(CTX-M-14) and bla(CTX-M-15) genes . A sequence similar to the inverted right repeat of ISEcp1 was identified downstream of bla(CTX-M-3), bla(CTX-M-10) and bla(CTX-M-15) genes suggesting the mobilization of these beta-lactamase genes by transposition events . In addition, Orf513 was identified upstream of the bla(CTX-M-2) gene . This work further underlined widespread of bla(CTX-M-15) gene associated with ISEcp1 . Spectrochim Acta A Mol Biomol Spectrosc, 2004 May, 60(6), 1279 - 89 Chromophoric spin-labeled beta-lactam antibiotics for ENDOR structural characterization of reaction intermediates of class A and class C beta-lactamases; Mustafi D et al.; The chromophoric spin-label substrate 6-N-{3-(2,2,5,5-tetramethyl-1-oxypyrrolin-3-yl)-propen-2-oyl}penicillanic acid (SLPPEN) was synthesized by acylation of 6-aminopenicillanic acid with the acid chloride of 3-(2,2,5,5-tetramethyl-1-oxypyrrolinyl)-2-propenoic acid and characterized by physical methods . By application of angle-selected electron nuclear double resonance (ENDOR), we have determined the molecular structure of SLPPEN in solution . SLPPEN exhibited UV absorption properties that allowed accurate monitoring of the kinetics of its enzyme-catalyzed hydrolysis . The maximum value of the (substrate-product) difference extinction coefficient was 2824 M(-1) cm(-1) at 275 nm compared to 670 M(-1) cm(-1) at 232 nm for SLPEN {J . Am . Chem . Soc . 117 (1995) 6739} . For SLPPEN, the steady-state kinetic parameters kcat and kcat/KM, determined under initial velocity conditions, were 637 +/- 36 s(-1) and 13.8 +/- 1.4 x 10(6) M(-1) s(-1), respectively, for hydrolysis catalyzed by TEM-1 beta-lactamase of E . coli, and 0.5 +/- 0.04 s(-1) and 3.9 +/- 0.4 x 10(4) M(-1) s(-1) for hydrolysis catalyzed by the beta-lactamase of Enterobacter cloacae P99 . We have also observed "burst kinetics" for the hydrolysis of SLPPEN with P99 beta-lactamase, indicative of formation of an acylenzyme reaction intermediate . In DMSO:H2O (30:70, v:v) cryosolvent mixtures buffered to pH* 7.0, the half-life of the acylenzyme intermediate formed with the P99 enzyme at -5 degrees C was > or = 3 min, suitable for optical characterization . The observation of burst kinetics in the hydrolysis of SLPPEN catalyzed by P99 beta-lactamase suggests that this chromophoric spin-labeled substrate is differentially sensitive to active site interactions underlying the cephalosporinase and penicillinase reactivity of this class C enzyme. Microbiology, 2004 May, 150(Pt 5), 1439 - 46 Unique organization and regulation of the mrx fimbrial operon in Xenorhabdus nematophila; He H et al.; Xenorhabdus nematophila, a Gram-negative bacterium belonging to the Proteus clade of the family Enterobacteriaceae, forms a mutualistic association with the soil nematode Steinernema carpocapsae . The nematode invades insects and releases Xenorhabdus into the haemolymph, where it participates in insect killing . To begin to understand the role of fimbriae in the unique life cycle of Xenorhabdus, the organization and expression of the mrx fimbrial operon was analysed . The mrx operon contained only five structural genes (mrxACDGH), making it one of the smallest chaperone-usher fimbrial operons studied to date . Unlike the mrp operon of Proteus mirabilis, a site-specific recombinase was not linked to the mrx operon . The intergenic region between the major fimbrial gene (mrxA) and the usher gene (mrxC) lacked a mrpB-like gene, but contained three tandem inverted repeat sequences located downstream of mrxA . A 940 nt mrxA-containing mRNA was the major transcript produced in cells growing on agar, while an mrx polycistronic mRNA was produced at low levels . A canonical sigma(70) promoter, identified upstream of mrxA, was not subject to promoter inversion . Fimbriae were not produced in an lrp-mutant strain, suggesting that the leucine-responsive regulatory protein, Lrp, plays a role in the regulation of the mrx operon . These findings show that the genetic organization and regulation of the mrx operon is in several respects distinct from other chaperone-usher fimbrial operons. J Biol Chem, 2004 Jul 16, 279(29), 30563 - 72 Epub 2004 May 05. Atomic resolution structures and solution behavior of enzyme-substrate complexes of Enterobacter cloacae PB2 pentaerythritol tetranitrate reductase . Multiple conformational states and implications for the mechanism of nitroaromatic explosive degradation; Khan H et al.; The structure of pentaerythritol tetranitrate (PETN) reductase in complex with the nitroaromatic substrate picric acid determined previously at 1.55 A resolution indicated additional electron density between the indole ring of residue Trp-102 and the nitro group at C-6 of picrate . The data suggested the presence of an unusual bond between substrate and the tryptophan side chain . Herein, we have extended the resolution of the PETN reductase-picric acid complex to 0.9 A . This high-resolution analysis indicates that the active site is partially occupied with picric acid and that the anomalous density seen in the original study is attributed to the population of multiple conformational states of Trp-102 and not a formal covalent bond between the indole ring of Trp-102 and picric acid . The significance of any interaction between Trp-102 and nitroaromatic substrates was probed further in solution and crystal complexes with wild-type and mutant (W102Y and W102F) enzymes . Unlike with wild-type enzyme, in the crystalline form picric acid was bound at full occupancy in the mutant enzymes, and there was no evidence for multiple conformations of active site residues . Solution studies indicate tighter binding of picric acid in the active sites of the W102Y and W102F enzymes . Mutation of Trp-102 does not impair significantly enzyme reduction by NADPH, but the kinetics of decay of the hydride-Meisenheimer complex are accelerated in the mutant enzymes . The data reveal that decay of the hydride-Meisenheimer complex is enzyme catalyzed and that the final distribution of reaction products for the mutant enzymes is substantially different from wild-type enzyme . Implications for the mechanism of high explosive degradation by PETN reductase are discussed. Appl Environ Microbiol, 2004 May, 70(5), 3013 - 23 Genomic diversity of Erwinia carotovora subsp . carotovora and its correlation with virulence; Yap MN et al.; We used genetic and biochemical methods to examine the genomic diversity of the enterobacterial plant pathogen Erwinia carotovora subsp . carotovora . The results obtained with each method showed that E . carotovora subsp . carotovora strains isolated from one ecological niche, potato plants, are surprisingly diverse compared to related pathogens . A comparison of 23 partial mdh sequences revealed a maximum pairwise difference of 10.49% and an average pairwise difference of 2.13%, values which are much greater than the maximum variation (1.81%) and average variation (0.75%) previously reported for Escherichia coli . Pulsed-field gel electrophoresis analysis of I-CeuI-digested genomic DNA revealed seven rrn operons in all E . carotovora subsp . carotovora strains examined except strain WPP17, which had only six copies . We identified 26 I-CeuI restriction fragment length polymorphism patterns and observed significant polymorphism in fragment sizes ranging from 100 to 450 kb for all strains . We detected large plasmids in two strains, including the model strain E . carotovora subsp . carotovora 71 . The two least virulent strains had an unusual chromosomal structure, suggesting that a particular pulsotype is correlated with virulence . To compare chromosomal organization of multiple enterobacterial genomes, several genes were mapped onto I-CeuI fragments . We identified portions of the genome that appear to be conserved across enterobacteria and portions that have undergone genome rearrangements . We found that the least virulent strain, WPP17, failed to oxidize cellobiose and was missing several hrp and hrc genes . The unexpected variability among isolates obtained from clonal hosts in one region and in one season suggests that factors other than the host plant, potato, drive the evolution of this common environmental bacterium and key plant pathogen. Appl Environ Microbiol, 2004 May, 70(5), 2959 - 65 Isolation, characterization, and U(VI)-reducing potential of a facultatively anaerobic, acid-resistant Bacterium from Low-pH, nitrate- and U(VI)-contaminated subsurface sediment and description of Salmonella subterranea sp . nov; Shelobolina ES et al.; A facultatively anaerobic, acid-resistant bacterium, designated strain FRCl, was isolated from a low-pH, nitrate- and U(VI)-contaminated subsurface sediment at site FW-024 at the Natural and Accelerated Bioremediation Research Field Research Center in Oak Ridge, Tenn . Strain FRCl was enriched at pH 4.5 in minimal medium with nitrate as the electron acceptor, hydrogen as the electron donor, and acetate as the carbon source . Clones with 16S ribosomal DNA (rDNA) sequences identical to the sequence of strain FRCl were also detected in a U(VI)-reducing enrichment culture derived from the same sediment . Cells of strain FRCl were gram-negative motile regular rods 2.0 to 3.4 micro m long and 0.7 to 0.9 microm in diameter . Strain FRCl was positive for indole production, by the methyl red test, and for ornithine decarboxylase; it was negative by the Voges-Proskauer test (for acetylmethylcarbinol production), for urea hydrolysis, for arginine dihydrolase, for lysine decarboxylase, for phenylalanine deaminase, for H(2)S production, and for gelatin hydrolysis . Strain FRCl was capable of using O(2), NO(3)(-), S(2)O(3)(2-), fumarate, and malate as terminal electron acceptors and of reducing U(VI) in the cell suspension . Analysis of the 16S rDNA sequence of the isolate indicated that this strain was 96.4% similar to Salmonella bongori and 96.3% similar to Enterobacter cloacae . Physiological and phylogenetic analyses suggested that strain FRCl belongs to the genus Salmonella and represents a new species, Salmonella subterranea sp . nov. Rev Saude Publica, 2004 Apr, 38(2), 326 - 8 Epub 2004 Apr 26. {Antibacterial activity of essential oils on microorganisms isolated from urinary tract infection}; Pereira RS et al.; The antibacterial activity of essential oils extracted from medicinal plants (Ocimum gratissimum, L., Cybopogum citratus (DC) Stapf., and Salvia officinalis, L.) was assessed on bacterial strains derived from 100 urine samples . Samples were taken from subjects diagnosed with urinary tract infection living in the community . Microorganisms were plated on Muller Hinton agar . Plant extracts were applied using a Steers replicator and petri dishes were incubated at 37 degrees C for 24 hours . Salvia officinalis, L . showed enhanced inhibitory activity compared to the other two herbs, with 100% efficiency against Klebsiella and Enterobacter species, 96% against Escherichia coli, 83% against Proteus mirabilis, and 75% against Morganella morganii. Int J Antimicrob Agents, 2004 May, 23(5), 506 - 9 Resistance of uropathogens in symptomatic urinary tract infections in León, Nicaragua; Matute AJ et al.; Management of urinary tract infections (UTI) in Central America and especially Nicaragua, is complicated by the lack of knowledge about the antibiotic resistance of uropathogens . We conducted a prevalence study to gain more insight into the aetiology, bacterial resistance and risk factors for symptomatic UTI in the region of Leon, Nicaragua . In 2002, all consecutive patients with UTI symptoms and pyuria >/=10 WBC/hpf were admitted to the study . Positive cultures from midstream urine specimens were defined as >/=10(5) cfu/ml of a single uropathogen . Susceptibility tests were performed with disc diffusion tests using the Kirby-Bauer method and broth microdilution using National Committee for Clinical Laboratory Standards criteria both in Leon and a reference laboratory in Utrecht . A positive culture was present in 62 of 208 study subjects (30%) . Escherichia coli (56%), Klebsiella spp . (18%) and Enterobacter spp . (11%) were the most frequent pathogens isolated . Presence of cystocele, incontinence and increasing age were risk factors for bacterial UTI . E . coli was least resistant to ceftriaxone, amikacin and nitrofurantoin (>90% susceptible) . We observed high resistance rates in E . coli to amoxicillin (82%, MIC(90) 128 mg/l), trimethoprim-sulphamethoxazole (TMP-SMX) (64%, MIC(90) 32 mg/l), cephalothin (58%, MIC(90), 32 mg/l), ciprofloxacin (30%; MIC(90), 32 mg/l), amoxicillin/clavulanate (21%, MIC(90) 8 mg/l) and gentamicin (12%, MIC(90) 2 mg/l) . Our results suggests that community acquired uropathogens in Nicaragua are highly resistant to many antimicrobial agents . The use of amoxicillin, trimethoprim-sulphamethoxazole and cephalothin against uropathogens needs to be reconsidered . High quinolone resistance rates among E . coli in Nicaragua gives cause for great concern. Int J Antimicrob Agents, 2004 May, 23(5), 480 - 6 Antimicrobial susceptibility of clinical isolates of Enterobacteriaceae producing complex beta-lactamase patterns including extended-spectrum enzymes; Segatore B et al.; The antimicrobial susceptibility of 103 clinical isolates of Enterobacteriaceae to 11 antibiotics, was investigated, using a conventional inoculum size (5 x 10(5) CFU) and a higher inoculum size (5 x 10(8) CFU) . All the isolates produced complex beta-lactamase patterns, including an extended-spectrum beta-lactamase (ESBL) of the TEM- or SHV-type plus other enzymes (a TEM-type or an SHV-type non-ESBL and/or a class C enzyme) . The following repertoire of ESBLs was produced by the isolates: TEM-15, TEM-19, TEM-26, TEM-52, TEM-72, TEM-87, TEM-92, SHV-2a, SHV-5 and SHV-12, as assessed by sequencing . Production of the other enzymes was showed by analytical isoelectric focusing . Overall, meropenem was the most active agent and less influenced by inoculum size, while other beta-lactams showed a lower activity and a significant inoculum size effect . In conclusion, from its in vitro performance, meropenem could be considered as the last resource drug against strains producing complex beta-lactamase patterns including an ESBL. J Endotoxin Res, 2004, 10(2), 107 - 12 Enzymology of lipid A palmitoylation in bacterial outer membranes; Bishop RE et al.; The enzymology of palmitate addition to lipid A can be traced to the early discovery of monosaccharide lipid A precursors, but the functional importance of lipid A palmitoylation in bacterial resistance to the host immune response has emerged only recently . Lipid A palmitoylation in enterobacteria is determined by a PhoP/PhoQ-activated gene pagP, which encodes an unusual outer membrane enzyme of lipid A biosynthesis . PagP structure and dynamics have now been elucidated by both NMR spectroscopy and X-ray crystallography . PagP is an 8-stranded antiparallel beta-barrel preceded by an N-terminal amphipathic alpha-helix . The PagP barrel axis is uniquely tilted by 30 degrees with respect to the membrane normal . An interior hydrophobic pocket in the upper half of the molecule functions as a hydrocarbon ruler, which allows the enzyme to distinguish palmitate from other acyl chains found in phospholipids . Internalization of a phospholipid palmitoyl group within the barrel appears to occur by lateral diffusion from the outer leaflet through non-hydrogen bonded regions between beta-strands . The MsbA-dependent trafficking of lipids from the inner membrane to the outer membrane outer leaflet is necessary for lipid A palmitoylation in vivo . Efforts to determine the PagP catalytic mechanism may lead to the development of inhibitors for the treatment of infections. Scand J Infect Dis, 2004, 36(3), 165 - 73 Gram-negative bacillary meningitis after cranial surgery or trauma in adults; Briggs S et al.; In order to assess the clinical features, aetiology, treatment and outcome of post-neurosurgical and post-traumatic Gram-negative bacillary meningitis (GNBM) we performed a retrospective review of all adult patients admitted to the Department of Neurosurgery who had Gram-negative bacilli cultured from cerebrospinal fluid (CSF) following a neurosurgical procedure or traumatic head/spinal injury . During the 12 y of the review 33 patients had CSF isolates of Gram-negative bacilli that were thought to be significant . The median patient age was 47 y (range 22-77 y) and 21 (64%) were male . Klebsiella pneumoniae, Enterobacter cloacae and Escherichia coli were the most common isolates . Minimal inhibitory concentrations (MIC) measured for half the patients' isolates resulted in 5 regimen changes, including 2 patients with E . cloacae meningitis in whom cephalosporin susceptibility decreased during cephalosporin treatment . Our recommended initial treatment was intravenous ceftriaxone and amikacin, subsequently tailored by susceptibility results; approximately half the patients remained on the antibiotics they started and half were changed to an alternate regimen, most often a carbapenem . Five patients (15%) died, 1 dying after cure of his GNBM . There were no failures in those who received more than 12 d of appropriate treatment: treatment for at least 14 d after the last positive CSF culture guaranteed cure . Initial ceftriaxone and amikacin subsequently changing to susceptibility driven alternatives, often a carbapenem, resulted in cure of 85% of our patients with GNBM. Jpn J Infect Dis, 2004 Apr, 57(2), 37 - 40 An epidemiological analysis of Stenotrophomonas maltophilia strains in a university hospital; Caylan R et al.; The aim of this investigation was to evaluate the epidemiology of Stenotrophomonas maltophilia in a university hospital of Turkey . From June 2000 to December 2001, S . maltophilia strains were collected, clinical presentations were noted, and MIC determinations were performed by means of E-test . Enterobacterial repetitive intergenic consensus sequences-PCR (ERIC-PCR) was used for molecular typing of the strains . Forty-four strains of S . maltophilia were isolated from 41 hospitalized patients in a teaching hospital . The majority of specimens were from the blood and respiratory tract . Antimicrobial sensitivities of these strains were as follows: 97.7 % trimethoprim-sulfamethoxazole, 15.9% ticarcillin, and 95.4% ticarcillin- clavulanate . The strains were evaluated using the ERIC-PCR method . It was of interest to note that epidemiological typing revealed three small outbreaks that were caused by a total of 12 strains . The remaining isolates generated singular DNA patterns . DNA amplification was possible in 38 isolates and yielded 26 different patterns in a period of 20 months, leading to the suggestion that commensal bacteria becomes selected in the presence of a suitable host. J Antimicrob Chemother, 2004 Jun, 53(6), 1090 - 4 Epub 2004 Apr 29. In vitro susceptibility of recent antibiotic-resistant urinary pathogens to ertapenem and 12 other antibiotics; Alhambra A et al.; Background: The treatment of complicated urinary tract infections may require the use of a parenteral antibiotic with potent activity against the most common urinary pathogens . Ertapenem is a broad-spectrum 1beta-methyl carbapenem with a long plasma half-life that allows administration of a single daily dose . METHODS: The purpose of this work was to test the in vitro susceptibility to ertapenem, ampicillin, cefazolin, cefuroxime, cefotaxime, co-amoxiclav, piperacillin/tazobactam, imipenem, gentamicin, amikacin, fosfomycin, ciprofloxacin and co-trimoxazole of 482 strains of urinary pathogens of the family Enterobacteriaceae isolated from patients in the community of Madrid (40% from males) . The distribution was as follows: Escherichia coli (n = 315), Proteus mirabilis (n = 42), Klebsiella spp . (n = 14) and AmpC-producing Enterobacteriaceae (n = 111) . The strains studied were selected based on their resistance to quinolones and aminoglycosides, and their production of extended-spectrum beta-lactamases (ESBLs) or AmpC-type beta-lactamases . RESULTS: All the strains were susceptible to ertapenem, imipenem and amikacin . The MIC(90) of ertapenem ranged from a minimum of 0.03 mg/L for Proteus vulgaris and a maximum of 1 mg/L for Enterobacter spp . Ertapenem was the most active of all drugs tested in all cases . On comparing antibiotic resistance among ESBL-producing strains of E . coli (n = 35) and E . coli strains not producing ESBLs (n = 280), statistically significant differences were obtained for ciprofloxacin (P = 0.002) and gentamicin (P = 0.011) . Regarding ertapenem, only a slight increase in MIC(50) was seen, the value being 0.015 mg/L for strains not producing ESBLs versus 0.03 mg/L for ESBL-producing strains . CONCLUSIONS: In view of its significant antibiotic potency against antibiotic-resistant Enterobacteriaceae, ertapenem may constitute a good therapeutic alternative in urinary infections caused by these pathogens. Am J Physiol Gastrointest Liver Physiol, 2004 Sep, 287(3), G638 - 46 Epub 2004 Apr 29. Muscularis inflammation and the loss of interstitial cells of Cajal in the endothelin ETB receptor null rat; Suzuki T et al.; Endothelin receptor null rats {ETB(-/-)} are a model for long-segment Hirschsprung's disease . These animals have significant intestinal distension (megaileum) proximal to a constricted region of the gastrointestinal tract lacking enteric ganglia . Experiments were performed to determine the pathophysiological changes that occur in these animals and to examine the tunica muscularis as a unique, immunologically active compartment . We observed abnormal intestinal flora in ETB(-/-) rats, which included a marked increase in gram-negative aerobes (Enterobacteriaceae) and anaerobes (Bacteroidaceae) in the distended region of the small intestine . Histochemical observations showed that neutrophilic infiltration was rarely or not observed, but the number of ED2 positive macrophages was increased in the tunica muscularis . Expression of IL-1beta and IL-6 mRNA was also significantly increased, and the level of CD14 (LPS receptors) were increased significantly in the tunica muscularis . Spontaneous phasic contractions were irregular in the distended intestinal regions of ETB(-/-) rats, and this was associated with an increased number of macrophages and damage to interstitial cells of Cajal (ICC) as revealed by using Kit-like immunoreactivity and electron microscopy . These results suggest that ED2-positive resident macrophages may play an important role in the inflammation of tunica muscularis in ETB(-/-) rats . Increased numbers and activation of macrophages may result in damage to ICC networks leading to disordered intestinal rhythmicity in regions of the gut in which myenteric ganglia are intact. Crit Rev Microbiol, 2004, 30(1), 25 - 32 Extended-spectrum beta-lactamases (ESbLs): characterization, epidemiology and detection; Shah AA et al.; Beta-lactamases of Enterobacteriaceae are the most important mechanism of resistance against beta-lactam drugs . Two types of beta-lactamases can confer resistance against 3rd generation cephalosporins . Chromosomally mediated beta-lactamases are inducible and are not inhibited by clavulanic acid . Resistance due to these enzymes is non-transferable . The 2nd type of enzyme is plasmid-mediated beta-lactamases, which are inhibited by clavulanic acid . These enzymes are more important clinically as these can be transferred between various species of Enterobacteriaceae . These enzymes are called extended-spectrum beta-lactamases (ESBLs) . ESBL-producing Enterobacteriaceae have been responsible for numerous outbreaks of infection throughout the world and pose challenging infection control issues . Antibacterial choice is often complicated by multi-resistance . ESBLs can confer resistance against all beta-lactam drugs except carbapenems and cephamycins . Nursing home patients may be an important reservoir of ESBL-containing multiple antibiotic-resistant organisms . Use of broad-spectrum oral antibiotics and probably poor infection control practices may facilitate spread of this plasmid-mediated resistance . In addition to known populations at risk, ambulatory patients with chronic conditions represent another patient population that may harbor ESBL-producing organisms . Various methods can be used for detection of ESBLs in the laboratory . These tests include double disc diffusion test, Vitek ESBL test, E Tests, MIC Determination, genetic method, and isoelectric focusing (IEF). Jpn J Antibiot, 2004 Feb, 57(1), 70 - 104 {Nationwide surveillance of parenteral antibiotics containing meropenem activities against clinically isolated strains in 2002}; Yamaguchi K et al.; The antibacterial activity of meropenem (MEPM) and other parenteral antibiotics against clinical isolates of 899 strains of Gram-positive bacteria, 1500 strains of Gram-negative bacteria, and 158 strains of anaerobic bacteria obtained from 28 medical institutions during 2002 was measured . The results were as follows; 1 . MEPM was more active than other carbapenem antibiotics against Gram-negative bacteria, especially against enterobacteriaceae and Haemophilus influenzae . MIC90 of MEPM against Pseudomonas aeruginosa was the lowest of the drugs tested . MEPM showed low cross-resistant rate against both imipenem-resistant P . aeruginosa and ciprofloxacin-resistant P . aeruginosa . MEPM was active against most of the species tested in Gram-positive and anaerobic bacteria, except for multi-drug resistant strains including methicillin-resistant Staphylococcus aureus (MRSA), methicillin-resistant Staphylococcus epidermidis (MRSE) . 2 . The proportion of extended-spectrum beta-lactamase (ESBL) strains was 3.1% (4 strains) in Escherichia coli and 1.9% (2 strains) in Klebsiella pneumoniae . Carbapenems including MEPM were active against these ESBL strains . In conclusion, the results from this surveillance study suggest that MEPM retains its potent and broad antibacterial activity and therefore is a clinically useful carbapenem; at present, 7 years after available for commercial use. Eur J Immunol, 2004 May, 34(5), 1461 - 71 Necrosis-like cell death induced by bacteria in mouse macrophages; Kirschnek S et al.; The death of individual cells is a frequent and physiological event in the mammalian immune system and most often occurs by apoptosis . It is becoming increasingly clear that cell death is also induced during bacterial infections . Here we report that, in addition to the apoptotic form already established, a necrosis-like form of cell death is induced by pyogenic bacteria (Enterobacteriaceae, Pseudomonas, enterococci) in mouse macrophages . Necrosis could be separated from apoptosis as it did not require phagocytosis of bacteria and occurred when apoptosis was inhibited by caspase blockade or by Bcl-2 . Furthermore, ligands that stimulate Toll-like receptors were also found to have the capacity to induce necrosis . Strikingly, this form of cell death was sufficient for the uptake of dead cells by either mouse bone marrow-derived DC or a cell line derived from DC, possibly by virtue of the externalization of phosphatidylserine . Since the loading with bacteria-carrying cells is likely to impact on DC function, this form of necrosis may have a previously unsuspected role in the development of an immune response. Clin Microbiol Infect, 2004 May, 10(5), 436 - 40 Prevalence of colonisation with third-generation cephalosporin-resistant Enterobacteriaceae in ICU patients of Heidelberg University Hospitals; von Baum H et al.; The aim of this study was to assess colonisation and transmission of third-generation cephalosporin-resistant Enterobacteriaceae (CRE) from patients in 16 intensive care units . A prospective, repetitive point prevalence survey was performed over 6 months, involving samples from 1851 patients . CRE were isolated from 186 (10%) patients, with Enterobacter spp . being the most common . Mean point prevalence rates were significantly higher for paediatric wards (22.5%) compared to surgical (8.1%) and medical (5.5%) units . All CRE isolates were typed by pulsed-field gel electrophoresis . Non-outbreak nosocomial transmission rates of these pathogens were calculated as 12.8% for paediatric patients, compared to 6.8% for adult patients, which may reflect differences in sensitivity to overgrowth with resistant bacteria and contact with health care workers. Clin Microbiol Infect, 2004 May, 10(5), 416 - 20 Evaluation of the OSIRIS video reader as an automated measurement system for the agar disk diffusion technique; Kolbert M et al.; Measurement of inhibition zones by the automated OSIRIS system was compared with manual measurement . In total, 14 176 measurements were made with 352 staphylococcal and 80 Enterobacteriaceae isolates, involving four panels of antibiotics on round and square Mueller-Hinton agar plates, according to the German DIN 58940 recommendations . Variations of +/- 3 mm in zone size measurements were defined as tolerable . Very major errors (i.e., classification of a resistant isolate as susceptible by the OSIRIS system) occurred in < 1% of tests . With staphylococci, the best concordance was recorded for rifampicin (91.3%), moxifloxacin (88.1%), and gentamicin (86.3%), while the concordance on square plates for vancomycin, pristinamycin and kanamycin was 97.2%, 96.1% and 96.0%, respectively . The poorest concordance was for cefuroxime (43.7%) and novobiocin (47.0%) on round plates, and fosfomycin (36.5%) and chloramphenicol (84.0%) on square plates . With Enterobacteriaceae, 100% concordance was recorded for ampicillin, gentamicin and ciprofloxacin on round agar plates, and for gentamicin, cefoxitin and nalidixic acid on square plates . The poorest results were recorded for nalidixic acid (32.5%) and piperacillin (82.5%) on round plates, and for nitrofurantoin (72.5%) and amoxycillin (82.5%) on square plates . It was concluded that the OSIRIS system was a rapid and reliable system for measuring disk susceptibility test results on round and square agar plates. Arch Biochem Biophys, 2004 May 15, 425(2), 184 - 92 Flavoenzyme-catalyzed redox cycling of hydroxylamino- and amino metabolites of 2,4,6-trinitrotoluene: implications for their cytotoxicity; Sarlauskas J et al.; The toxicity of 2,4,6-trinitrotoluene (TNT), a widespread environmental contaminant, is exerted through its enzymatic redox cycling and/or covalent binding of its reduction products to proteins and DNA . In this study, we examined the possibility of another cytotoxicity mechanism of the amino- and hydroxylamino metabolites of TNT, their flavoenzyme-catalyzed redox cycling . The above compounds acted as redox-cycling substrates for single-electron transferring NADPH:cytochrome P-450 reductase (P-450R) and ferredoxin:NADP(+) reductase (FNR), as well as substrates for the two-electron transferring flavoenzymes rat liver NAD(P)H:quinone oxidoreductase (NQO1) and Enterobacter cloacae NAD(P)H:nitroreductase (NR) . Their reactivity in P-450R-, FNR-, and NR-catalyzed reactions increased with an increase in their single-electron reduction potential (E(1)(7)) or the decrease in the enthalpy of free radical formation . The cytotoxicity of the amino- and hydroxylamino metabolites of TNT towards bovine leukemia virus-transformed lamb kidney fibroblasts (line FLK) was partly prevented by the antioxidant N,N'-diphenyl-p-phenylene diamine and desferrioxamine, and potentiated by 1,3-bis-(2-chloroethyl)-1-nitrosourea, thus pointing to the involvement of oxidative stress . In general, their cytotoxicity increased with an increase in their electron accepting properties, or their reactivity towards the single-electron transferring FNR and P-450R . Thus, our data imply that the flavoenzyme-catalyzed redox cycling of amino and hydroxylamino metabolites of TNT may be an important factor in their cytotoxicity. Int J Food Microbiol, 2004 Apr 15, 92(2), 217 - 25 Microbial contamination on beef in relation to hygiene assessment based on criteria used in EU Decision 2001/471/EC; McEvoy JM et al.; Thirty-six carcasses were sampled over a 12-month period at an Irish beef abattoir . Between one and five carcass sites (including the hock, brisket, cranial back, bung, inside round and outside round) were sampled after hind leg skinning, hide removal, bung tying, evisceration, splitting, washing, chilling for 24 h and boning, using a wet and dry, cotton wool swab technique . For each sample, total viable counts (TVC), Escherichia coli, total coliforms and Enterobacteriaceae were enumerated . The results are considered in relation to European Union Decision 2001/471/EC which sets performance criteria for TVCs and Enterobacteriaceae in samples taken by excision . Though not explicitly stated in the Decision, it has been proposed that microbiological performance criteria for samples taken by swabbing be set at 20% of the values set for excision samples . Therefore, log mean TVCs in carcass swab samples taken before chilling are acceptable, marginal and unacceptable when they are <2.8, 2.8-4.30 and >4.30 cm(-2), respectively . By these criteria, TVCs on carcasses in the present study were in the marginal range . The marginal result for TVCs was due in the most part to hide removal operations, particularly at the hock and brisket sites . Bacterial contamination on post-chill carcasses was similar or lower to that on pre-chill carcasses, while boning resulted in general increases in TVCs and in E . coli, total coliform and Enterobacteriaceae numbers . In Decision 2001/471/EC, the effects of chilling and boning are not included in the assessment of process control . Data from this study indicate that performance criteria based on log mean Enterobacteriaceae values are unsuitable because of the infrequent occurrence of these organisms on the carcass. Int J Food Microbiol, 2004 Apr 15, 92(2), 181 - 7 Antimicrobial and antioxidative enrichment of oak (Quercus robur) bark by rotation planar extraction using ExtraChrom; Andrensek S et al.; The multifunctional ExtraChrom instrument was used in the extraction of antimicrobial and radical scavenging components from oak (Quercus robur L.) bark . Milled and sieved oak bark was extracted with 80% (v/v) methanol solution in water on the ExtraChrom instrument using step-gradient in the preparative separation . Extracts were tested using agar diffusion method on Staphylococcus aureus, Enterobacter aerogenes and Candida albicans . Some extracts showed moderate bactericidal, fungicidal, bacteriostatic and fungistatic activity . The composition related to activity of the fractions and extracts was screened simultaneously by thin-layer chromatography (TLC) detected by UV and by spraying the plate with radical scavenging reagent 1,1-diphenyl-2-picrylhydrazyl (DPPH) to detect antioxidant activity . Thus, we could demonstrate the antiradical and antimicrobial activity of oak beneficial in the storage of wine against the oxidation and human microbial exposure. Proc Natl Acad Sci U S A, 2004 May 4, 101(18), 7118 - 22 Epub 2004 Apr 23. Gram-positive bacteria are a major reservoir of Class 1 antibiotic resistance integrons in poultry litter; Nandi S et al.; Reversing the spread of antibiotic multiresistant bacteria is hampered by ignorance of the natural history of resistance genes, the mobile elements carrying them, and the bacterial hosts harboring them . Using traditional cultivation and cultivation-independent molecular techniques, we quantified antibiotic resistance genes and mobile elements called integrons in poultry house litter from commercial poultry farms . Unexpectedly, the major reservoir for Class 1 integrons in poultry litter is not their previously identified hosts, Gram-negative Enterobacteriaceae such as Escherichia coli . Rather, integrons and associated resistance genes abound in several genera of Gram-positive bacteria that constitute >85% of the litter community compared with Enterobacteriaceae that comprise <2% of this ecosystem . This finding warrants reexamination of our assumptions about the persistence and spread of antibiotic resistance genes. Presse Med, 2004 Apr 10, 33(7), 460 - 6 {Etiology of bacterial infections in febrile neutropenic patients: the role of the laboratory in the diagnosis}; Poyart C et al.; EPIDEMIOLOGICAL EVOLUTION: Until the mid-eighties, infectious complications (pneumonia, septicemia) observed in neutropenic patients were, in 70% of cases, of bacterial origin with Gram negative bacillae (Escherichia coli, Klebsiella sp, Pseudomonas aeruginosa) isolated 8 times out of 10 . Among the Gram positive bacteria, Staphylococcus aureus predominated . The etiological profile of bacterial infections has since evolved with a predominance (60 to 70%) of Gram positive bacteria (coagulase-negative staphylococci, viridans streptococci) and a change in the epidemiology of the Gram positive bacteria notably with a lesser frequency of P . aeruginosa infections . THE GRAM POSITIVE BACTERIA: Coagulase-negative staphylococci are among the first germs responsible for nosocomial bacteremia (central venous catheters) and they are usually multiresistant . Viridans streptococci are a frequent cause of bacteremia; they are generally sensitive to antibiotics active on Gram positive bacteria, but the incidence of resistant strains is increasing . Enterococci are in majority responsible for colonisation in neutropenic patients and less frequently for infections; they raise the problem of resistance to antibiotics, notably to glycopeptides . Other Gram positive bacteria can be responsible for infections in neutropenic patients; it is crucial that they be identified because they require treatment with an appropriate antibiotic . GRAM NEGATIVE BACTERIA: Among the enterobacteria, Escherichia coli is predominantly isolated and raises the problem of the increasing incidence of resistance to fluoroquinolone . Pseudomonas aeruginosa, less frequently responsible today, remains associated with a far greater rate of mortality than that observed with the other microorganisms . Other Gram negative bacteria can be identified; they require an adapted antibiotherapy because they are often naturally multiresistant to antibiotics . THE ROLE OF THE LABORATORY: For the diagnosis of infections in neutropenic patients, the microbiology laboratory has a determinating role . The laboratory ensures the analysis of various biological examinations: blood cultures, methods permitting the diagnosis of an infection on a permanent catheter, copro-cultures (research for common enteropathogens, quantification in the case of digestive decontamination, screening for multiresistant bacteria), cytobacteriological examination of urine, samples of respiratory origin, cytobacteriological examination of cerebro-spinal fluid...). Antimicrob Agents Chemother, 2004 May, 48(5), 1719 - 26 Peritoneal fluid titer test for peritoneal dialysis-related peritonitis; Strijack C et al.; Standard microbiological tests (i.e., MIC) do not account for the unique factors of peritoneal dialysis (PD)-related peritonitis which can significantly influence treatment response . Our goals were to develop a peritoneal fluid titer (PFT) test and to conduct a pilot study of its association with clinical outcome . The methodology was developed by using spent dialysate collected from patients with bacterial PD-related peritonitis prior to the initiation of antibiotics . Dialysate was processed and spiked with antibiotic to simulate two standard intraperitoneal regimens: cefazolin plus tobramycin and cefazolin alone . Thirty-six clinical isolates, including Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomonas aeruginosa, were tested . In the pilot study, dialysate was collected from 14 patients with bacterial PD-related peritonitis . Titers were determined by using each patient's dialysate and infecting pathogen . Titers were highly reproducible, with discrepancies in only 1% of cases . Overall, PFTs were notably higher against gram-positive bacteria (P < 0.0001) . The addition of tobramycin increased titers significantly from zero to values of 1/16 to 1/64 against E . cloacae and P . aeruginosa (P < 0.0001) . In the pilot study, peritoneal fluid inhibitory titers were significantly associated with clinical outcome, with a median value of 1/96 for patients who were cured compared to 1/32 for those who failed treatment (P = 0.036) . In conclusion, this study provides preliminary support for the PFT as a pharmacodynamic index specific to the treatment of PD-related peritonitis . With further characterization and validation in patients, the PFT test may advance the study of antibiotic therapies for PD-related peritonitis. Antimicrob Agents Chemother, 2004 May, 48(5), 1688 - 98 Comparative study of the effects of ceftizoxime, piperacillin, and piperacillin-tazobactam concentrations on antibacterial activity and selection of antibiotic-resistant mutants of Enterobacter cloacae and Bacteroides fragilis in vitro and in vivo in mixed-infection abscesses; Stearne LE et al.; The effects of ceftizoxime (CZX), piperacillin (PIP), and PIP-tazobactam (PT) concentrations on the antibacterial activity and selection of resistant mutants of Bacteroides fragilis and Enterobacter cloacae were investigated in vitro in a mixed-culture anaerobic time-kill study and in vivo in a mixed-infection abscess model . Mixed cultures were incubated for 24 h with 0.125 to 512 micro g of CZX per ml or 0.125 to 2,048 micro g of PIP or PT per ml . Mice were treated every 2 h for 24 h with CZX at 6 to 1,536 mg/kg/day or with PIP or PT at 24 to 6,144 mg/kg/day starting 30 min before inoculation with different B . fragilis-E . cloacae combinations . There was a good correlation between the in vitro and in vivo activities of the antibiotics and their MICs obtained with high inocula (10(8) CFU/ml) . The respective 50% effective doses (milligrams per kilogram per day) with B . fragilis and E . cloacae 22491 were 771 and 521 for CZX, 416 and 643 for PIP, and 85 and 554 for PT, and with the B . fragilis-E . cloacae 032349 combination, they were 81 and 21 for CZX and 77 and 766 for PT . Resistant mutants of E . cloacae 22491 were preferentially selected in vitro with 2 to 64 micro g of CZX per ml and in vivo with CZX at 12 to 384 mg/kg/day . There was no preferential selection of CZX-resistant B . fragilis or E . cloacae 032349 . For CZX-resistant E . cloacae 22491, we found a 16- to 512-fold increase in the MIC of CZX and increased MICs of other expanded-spectrum cephalosporins, owing in part to the production of a stably derepressed cephalosporinase . In vitro and in vivo, PT did not select resistant mutants of E . cloacae and B . fragilis . Results demonstrate the adverse microbiological outcome of choosing an expanded-spectrum cephalosporin like CZX for empirical treatment of mixed infections involving a susceptible Enterobacter strain. Med Dosw Mikrobiol, 2003, 55(4), 351 - 6 {Antibiotic sensitivity of Enterobacteriaceae isolated from women vagina and uterine cervix}; Nolewajka-Lasak I et al.; Based on performed investigation in the group of 200 women treated for recurrent and chronic vaginitis and cervicitis, the characteristic of isolated microorganism was done . There were found series of drug-resistant bacteria in the vagina and uterine cervical canal in women with recurrent vaginitis and chronic cervicitis . In 42.6% of patients with diagnosed chronic cervicitis (Cervicitis chronica n = 50) Escherichia coli strains were isolated, 5% of which produced extended spectrum beta-lactamases (ES beta L) . In women with recurrent vaginitis (Colpitis recidivans n = 150) Escherichia coli strains where isolated in 45.6% . Among them 4.5% produced ES beta L . Expression of beta-lactamases with broadened substrate spectrum was done in double-disc-test . CONCLUSIONS: 1 . Study of antibiotic sensitivity of isolated bacteria should be a diagnostic standard in bacterial infections of uterine cervix and vagina . 2 . The choice of appropriate antibiotics should take into consideration the drug-resistance mechanisms of isolated bacteria . 3 . When drug-resistant bacteria are isolated, combined therapy should be applied. Med Dosw Mikrobiol, 2003, 55(4), 343 - 9 {Evaluation of relations between plasmids and phage host range among clinical isolates of Enterobacter cloacae}; Nieradko J et al.; The aim of this study was evaluation the plasmid influence on phage host range of clinical strains of Enterobacter cloacae . We found that strains included in restrictive pattern A, displayed reduced host range . Such reduced sensitivity make these strains excellent candidates for search restrictive-modification systems . High discriminative efficacy of isolated phages (specific for strains Enterobacter cloacae) make them useful tool for phage typing in epidemiological investigations. Phytother Res, 2004 Mar, 18(3), 208 - 11 Antimicrobial effects of Quercus ilex L . extract; Gulluce M et al.; The antimicrobial activities of the methanol extract of Quercus ilex L . (Pirnal oak) leaves were tested in vitro against a wide range of human and plant-associated microorganisms . A total of 132 microbial organisms belonging to 55 bacteria and five fungi and yeast species were studied using a disc-diffusion method and microdilution assays . The results were evaluated as inhibition zones around the disc impregnated with Q . ilex extract at a concentration of 300 micro L/mL . The results showed that Q . ilex did not have any antifungal activities against Alterneria alternata, Aspergillus flavus, Fusarium oxysporum, Penicillum spp., whereas there were inhibition effects on the growth of all Candida albicans isolates . In total 97 bacterial strains (74%) were found to be resistant to Q . ilex extract . The remaining 35 (27%) strains of seven different bacteria genera including Brucella, Bacillus, Enterobacter, Neisseria, Pseudomonas and Escherichia were susceptible to the extract tested . The minimum inhibitory concentrations (MIC) of the extract ranged from 125 to 500 micro L/mL . These results suggest that Q . ilex possesses compounds with antibacterial and anticandidal properties . Ann Acad Med Singapore, 2004 Mar, 33(2), 228 - 34 Septic arthritis after arthroscopic anterior cruciate ligament reconstruction; Fong SY et al.; INTRODUCTION: A retrospective review of postoperative infected anterior cruciate ligament (ACL) reconstruction was done on 472 consecutive cases in one institution . The purpose was to assess the incidence, diagnosis, treatment and outcome factors . MATERIALS AND METHODS: Out of 472 arthroscopic-assisted ACL reconstructions performed between 1999 and 2002, 7 (1%) postoperative deep intra-articular infections were detected . Seven males with a mean age of 23 years (range, 19 to 30 years) formed the study group; 3 had undergone prior knee surgery . RESULTS: Four patients had acute infection (<2 weeks), 3 had subacute infection (2 weeks to 2 months) and none had late infection (>2 months) . All were admitted within 24 hours of onset of symptoms and underwent immediate arthroscopic lavage, incision and drainage of abscess, debridement with graft retention and intravenous (8 to 31 days) followed by oral (4 to 6 weeks) antibiotics . Staphylococcus aureus was present in 4 patients, Peptostreptococcus in 3, Klebsiella in 1, and Enterobacter in 1 . The patients underwent an average of 1.4 arthroscopic procedures (range, 1 to 3 procedures), with an average hospital stay of 17.3 days per patient . All were evaluated at an average of 11.7 months (range, 5 to 26 months) . In all cases, the infection resolved with stable knees and with all grafts and implants retained . Although rare, early diagnosis and prompt treatment of infection can result in successful eradication without sacrificing the graft. Environ Pollut, 1987, 48(4), 311 - 9 Bacterial bioabsorption of nickel from industrial cooling water; Kasan HC et al.; Three bacterial strains tolerant to the presence of 100 mg litre(-1) nickel ions were isolated from a water reclamation system . Each organism was tested for ability to accumulate nickel at the above-mentioned concentration . The organism capable of maximum nickel accumulation was identified as an Enterobacter sp . and intracellular nickel deposition by this microorganism was determined using energy dispersive X-ray analysis (EDAX) and transmission electron microscopy . A metal-staining technique for light microscopy was developed . Further studies revealed that growth and glucose utilisation by this isolate was inhibited in culture by nickel. Environ Pollut, 1990, 65(1), 1 - 17 Crude oil and hydrocarbon-degrading strains of Rhodococcus rhodochrous isolated from soil and marine environments in Kuwait; Sorkhoh NA et al.; Soil and marine samples collected from different localities in Kuwait were screened for microorganisms capable of oil degradation . Both fungi and bacteria were isolated . The fungal flora consisted of Aspergillus terreus, A . sulphureus, Mucor globosus, Fusarium sp . and Penicillum citrinum . Mucor globosus was the most active oil degrading fungus isolated . Bacterial isolates included Bacillus spp . Enterobacteriaceae, Pseudomonas spp., Nocardia spp., Streptomyces spp.,and Rhodococcus spp . Among these Rhodococcus strains were the most efficient in oil degradation and, relatively speaking, the most abundant . Bacterial and fungal isolates differed in their ability to degrade crude oil, with Rhodococcus isolates being more active that fungin in n-alkane biodegradation, particularly in the case of R . rhodochrous . In addition to medium chain n-alkanes, fungi utilized one or more of the aromatic hydrocarbons studied, while bacteria failed to do so . R . rhodochorous KUCC 8801 was shown by GLC and post-growth studies to be more efficient in oil degradation than isolates known to be active oil degraders. Am J Med Sci, 2004 Mar, 327(3), 118 - 22 Simultaneous presence of blaTEM and blaSHV genes on a large conjugative plasmid carried by extended-spectrum beta-lactamase-producing Klebsiella pneumoniae; Araque M et al.; BACKGROUND: Among members of the family Enterobacteriaceae, the production of plasmid-mediated extended-spectrum beta-lactamases (ESbetaLs) has emerged as an important mechanism of the resistance to beta-lactams . METHODS: The molecular basis of extended-spectrum beta-lactamase in 48 strains of Klebsiella pneumoniae, recovered from neonatal patients with nosocomial septicemia during an outbreak that occurred in November 2000 at a Neonatal Intensive Care Unit of The Andes University Hospital in Venezuela, were investigated . RESULTS: The isolates were resistant to expanded-spectrum cephalosporins, aztreonam, gentamicin, kanamycin, tetracycline, and chloramphenicol, but remained susceptible to cefoxitin, imipenem, amikacin, and tobramycin . Production of ESbetaL activity was confirmed by restoring susceptibility to ceftazidime in the presence of clavulanic acid . All isolates harbored an 87-kilobase plasmid . Analysis of the outer membrane protein patterns did not reveal relevant changes of the porins profile . All resistance markers were transferable to Escherichia coli by conjugation and were lost en bloc after treatment with acridine orange . Isoelectric focusing for beta-lactamases was performed on transconjugants, obtaining 2 bands with isoelectric points of 5.4 and 8.2 . Genes encoding both enzymes are located on the single, self-transferable 87-kilobase plasmid pKAM542 . Analysis of the plasmid by hybridization revealed the presence of both blaTEM and blaSHV determinants . Cloning and sequencing identified them as blaTEM-1 and blaSHV-5, respectively; the latter was responsible for the ESbetaL activity among nosocomial isolates of K pneumoniae . CONCLUSIONS: Microbiologists, epidemiologists, and clinicians must be aware of potential ESbetaL-encoding organisms to assess proper antimicrobial managements and effective infection controls. J Biol Chem, 2004 Jul 2, 279(27), 27928 - 40 Epub 2004 Apr 15. Biosynthesis of a novel 3-deoxy-D-manno-oct-2-ulosonic acid-containing outer core oligosaccharide in the lipopolysaccharide of Klebsiella pneumoniae; Frirdich E et al.; The core oligosaccharide region of Klebsiella pneumoniae lipopolysaccharide contains some novel features that distinguish it from the corresponding lipopolysaccharide region in other members of the Enterobacteriaceae family, such as Escherichia coli and Salmonella . The conserved Klebsiella outer core contains the unusual trisaccharide 3-deoxy-d-manno-oct-2-ulosonic acid (Kdo)-(2,6)-GlcN-(1,4)-GalUA . In general, Kdo residues are normally found in the inner core, but in K . pneumoniae, this Kdo residue provides the ligation site for O polysaccharide . The outer core Kdo residue can also be non-stoichiometrically substituted with an l-glycero-d-manno-heptopyranose (Hep) residue, another component more frequently found in the inner core . To understand the genetics and biosynthesis of core oligosaccharide synthesis in Klebsiella, the gene products involved in the addition of the outer core GlcN (WabH), Kdo (WabI), and Hep (WabJ) residues as well as the inner core HepIII residue (WaaQ) were identified . Non-polar mutations were created in each of the genes, and the resulting mutant lipopolysaccharide was analyzed by mass spectrometry . The in vitro glycosyltransferase activity of WabI and WabH was verified . WabI transferred a Kdo residue from CMP-Kdo onto the acceptor lipopolysaccharide . The activated precursor required for GlcN addition has not been identified . However, lysates overexpressing WabH were able to transfer a GlcNAc residue from UDP-GlcNAc onto the acceptor GalUA residue in the outer core. J Bacteriol, 2004 May, 186(9), 2540 - 7 Characterization of the alternative sigma factor sigma54 and the transcriptional regulator FleQ of Legionella pneumophila, which are both involved in the regulation cascade of flagellar gene expression; Jacobi S et al.; We cloned and analyzed Legionella pneumophila Corby homologs of rpoN (encoding sigma(54)) and fleQ (encoding sigma(54) activator protein) . Two other genes (fleR and pilR) whose products have a sigma(54) interaction domain were identified in the genome sequence of L . pneumophila . An rpoN mutant strain was nonflagellated and expressed very small amounts of the FlaA (flagellin) protein . Like the rpoN mutant, the fleQ mutant strain of L . pneumophila was also nonflagellated and expressed only small amounts of FlaA protein compared to the amounts expressed by the wild type . In this paper we show that the sigma(54) factor and the FleQ protein are involved in regulation of flagellar gene operons in L . pneumophila . RpoN and FleQ positively regulate the transcription of FliM and FleN, both of which have a sigma(54)-dependent promoter consensus sequence . However, they seemed to be dispensable for transcription of flaA, fliA, or icmR . Our results confirmed a recently described model of the flagellar gene regulation cascade in L . pneumophila (K . Heuner and M . Steinert, Int . J . Med . Microbiol . 293:133-145, 2003) . Flagellar gene regulation was found to be different from that of Enterobacteriaceae but seems to be comparable to that described for Pseudomonas or Vibrio spp. Di Yi Jun Yi Da Xue Xue Bao, 2004 Apr, 24(4), 439 - 40, 444 {Drug-resistance analysis of Gram-negative bacilli producing extended-spectrum beta-lactamases in lower respiratory tract infection}; Long J et al.; OBJECTIVE: To analyze occurrence of Gram-negative bacteria producing extended-spectrum beta-lactamase (ESBL) in patients with lower respiratory tract infection and assess their drug sensitivity to provide evidence for clinical management of lower respiratory tract infection with drug-resistant ESBL-producing bacteria . METHODS: A total of 312 Gram-negative bacterial strains were isolated from patients with lower respiratory tract infection and identified for the presence of ESBL by Vitek ESBL detection test and double-disk synergy tests as well as standard disk diffusion test . The effects of 10 antibacterial drugs on ESBL-producing strains were compared . RESULTS: A rate of 20.1% of the isolated strains were identified to produce ESBLs, including Klebsiella Pneumoniae, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginasa and Klebsiella oxytocla (accounting for 30.1%, 29.1%, 27.0%, 11.3%, and 2.5%, respectively, of the ESBL-producing strains) . Imipenem possessed the most powerful antibacterial activity against ESBL-producing strains . CONCLUSIONS: Attention should be given to lower respiratory tract infections with ESBL-producing Gram-negative bacilli, and imipenem can be the primary choice for fighting such infections. Cancer Res, 2004 Apr 15, 64(8), 2853 - 7 Efficient cancer therapy with a nanobody-based conjugate; Cortez-Retamozo V et al.; Nanobodies are the smallest fragments of naturally occurring single-domain antibodies that have evolved to be fully functional in the absence of a light chain . Nanobodies are strictly monomeric, very stable, and highly soluble entities . We identified a nanobody with subnanomolar affinity for the human tumor-associated carcinoembryonic antigen . This nanobody was conjugated to Enterobacter cloacae beta-lactamase, and its site-selective anticancer prodrug activation capacity was evaluated . The conjugate was readily purified in high yields without aggregation or loss of functionality of the constituents . In vitro experiments showed that the nanobody-enzyme conjugate effectively activated the release of phenylenediamine mustard from the cephalosporin nitrogen mustard prodrug 7-(4-carboxybutanamido) cephalosporin mustard at the surface of carcinoembryonic antigen-expressing LS174T cancer cells . In vivo studies demonstrated that the conjugate had an excellent biodistribution profile and induced regressions and cures of established tumor xenografts . The easy generation and manufacturing yield of nanobody-based conjugates together with their potent antitumor activity make nanobodies promising vehicles for new generation cancer therapeutics. Cir Cir, 2004 Jan-Feb, 72(1), 41 - 6 {Nosocomial infection surveillance: experience at a cardiology hospital in Mexico}; Valenzuela-Flores AA et al.; OBJECTIVE: To describe the results of the Nosocomial Infection surveillance program at the Cardiology Hospital in Centro Medico Nacional Siglo XXI of the Mexican Social Security Institute . METHODOLOGY: To inform of the epidemiologic follow-up results from January 2000 to July 2003 . Global frequency, infection rates by infection site, hospital services, and frequency of most common microorganisms were estimated . RESULTS: During this period, global incidence showed that in every 100 discharges, there were 4.3 infections (4.3/100) . Frequency of infection was slightly higher in the surgical intensive care unit . Average infection rate in lower respiratory tract infections was 27/1,000; in surgical-site infections: 8/1,000 (mediastinitis 0.8/1,000); it was found that in urinary tract infection, rate was 6.6/1,000 . Most commonly isolated microorganisms were: coagulase-negative Staphylococcus (25%), Enterobacter sp . (17%), Candida albicans (13%), S . aureus (9%), P . aeruginosa (9%) and K . pneumoniae (6%) . CONCLUSION: This study describes the epidemiology of nosocomial infections in a Cardiology Hospital . The information is obtained through epidemiologic follow-up programs . This information is very important to develop specific strategies for control of infections. Bacteriol Virusol Parazitol Epidemiol, 2002 Jul-Dec, 47(3-4), 185 - 91 {Beta-lactam resistance in aquatic Enterobacter cloacae strains using phenotypic and genotypic criteria}; Lazar V et al.; Bacterial resistance by producing of beta-lactamases represents an increasing problem of infections chemotherapy . beta-lactam hydrolyzing activities are detected in virtually all bacteria, from witch Enterobacter cloacae produce chromosomal beta-lactamases included in inducible class AmpC beta-lactamases . The purpose of this preliminary study was to investigate the antibiotic susceptibility of 7 inducible beta-lactamase-producing Enterobacter cloacae strains isolated from aquatic sources (river and polluted waters) . The identification to the species level was performed with the API 32E system and susceptibility to antimicrobial agents was tested by the disk diffusion method according to NCCLS recommendations . The following antibiotics were tested: ampicillin, amoxycillin/clavulanic acid, ceftazidime, cefotaxime, cephalotin, cefamandole, cephaclor, imipenem, amikacin, gentamycin, kanamycin, tobramycin, ciprofloxacin, norfloxacin, ofloxacin, nalidixic acid, tetracycline and chloramphenicol . Minimum inhibitory concentrations (MICs) were performed using dilution method in Mueller-Hinton broth with a 0.06-64 micrograms/ml concentration range for all antimicrobials and bacterial inoculum of about 1-2 x 10(8) cfu/ml . For the data analysis NCCLS breakpoints for resistance and sensitivity were used . Interaction of beta-lactamase inhibitor clavulanate with cefotaxime was performed by double-disk synergy test . Detection of inducible beta-lactamase expression was performed by the inductibility disk diffusion test using cefotaxime, ceftazidime and imipenem . Genomic DNA was isolated using CTAB technique and bacterial plasmid isolation was performed by an alkaline lysis method . Genetic characterization was performed by agarose gel electrophoresis and spectrophotometric analysis . The majority of examined E . cloacae strains were sensitive to imipenem, cefamandole, amikacin and quinolones (norfloxacin and ofloxacin), a higher moderate resistance being observed only to nalidixic acid (higher than 50%) and ciprofloxacin (15%) . The percentage of resistant strains ranged from 72% (to kanamycin) to 87% (to gentamicin) . The best results (resistance about 99%) were obtained with ampicillin, amoxycillin/clavulanic acid, ceftazidime, cefotaxime, cephalotin, cephaclor, tobramycin, tetracycline and chloramphenicol . The disk diffusion tests showed the absence of extended-spectrum beta-lactamases production and the expression of inducible beta-lactamases . Electrophoretic patterns point out the presence of plasmid DNA . Plasmid profile revealed the presence of several different plasmids ranging from 2.5 kpb to more than 30 kpb . The presence of inducible beta-lactamase E . cloacae strains in aquatic media (river and polluted waters) and the closely related pattern of susceptibility among these strains reflect the possible contamination of these sources and the common origin of them. Bacteriol Virusol Parazitol Epidemiol, 2002 Jul-Dec, 47(3-4), 137 - 42 {Enterobacteria isolated from different pathological products and their sensitivity to antibiotics}; Idomir M et al.; The objectives of the study we have enhanced have focused on identifying the etiological implication of the Enterobacteria isolated from different pathological products and determining their sensitivity to the main antibiotics . We observed that from pharyngeal exudate and sputum the Enterobacter spp . strains were isolated the most frequently while from purulent secretion, urine, puncture liquids and blood, E . coli strains were the most frequently implicated . For all the categories etiologically implicated, there were selected strains resistant to one or more of the antibiotics tested, showing once again the importance of antibiogram in the initiation and control of infections disease treatment. J Food Prot, 2004 Apr, 67(4), 658 - 65 Escherichia coli O157 prevalence and enumeration of aerobic bacteria, Enterobacteriaceae, and Escherichia coli O157 at various steps in commercial beef processing plants; Arthur TM et al.; The effectiveness of current antimicrobial interventions used in reducing the prevalence or load of Escherichia coli O157 and indicator organisms on cattle hides and carcasses at two commercial beef processing plants was evaluated . Sponge sampling of beef cattle was performed at five locations from the initial entry of the animals to the slaughter floor to the exit of carcasses from the "hotbox" cooler . For each sample, E . coli O157 prevalence was determined and total aerobic bacteria, Enterobacteriaceae, and E . coli O157 were enumerated . E . coli O157 was found on 76% of animal hides coming into the plants, but no carcasses leaving the cooler were identified as contaminated with E . coli O157 . A positive relationship was seen between the incidence of E . coli O157 in hide samples and that in preevisceration samples . Aerobic plate counts and Enterobacteriaceae counts averaged 7.8 and 6.2 log CFU/100 cm2, respectively, on hides, and 1.4 and 0.4 log CFU/100 cm2, respectively, on chilled carcasses . Aerobic plate counts and Enterobacteriaceae counts on preevisceration carcasses were significantly related to the respective levels on the corresponding hides; the carcasses of animals whose hides carried higher numbers of bacteria were more likely to carry higher numbers of bacteria . Implementation of the sampling protocol described here would allow processors to evaluate the efficacy of on-line antimicrobial interventions and allow industrywide benchmarking of hygienic practices. J Food Prot, 2004 Apr, 67(4), 646 - 50 Prevalence of Escherichia coli O157 and levels of aerobic bacteria and Enterobacteriaceae are reduced when hides are washed and treated with cetylpyridinium chloride at a commercial beef processing plant; Bosilevac JM et al.; The objective of this experiment was to test the potential of a combined water wash and cetylpyridinium chloride (CPC) treatment as a hide intervention applied to cattle in the holding pens of a processing plant immediately before stunning . Over 2 processing days, 149 control and 139 treated cattle were tested . Control cattle were processed in the normal manner . The treatment group was prewashed with water the day before harvest . Immediately before stunning, these cattle were sprayed twice with 1% CPC, first for 3 min, then for 1 min . Hides and preevisceration carcasses were sampled to determine aerobic plate counts, Enterobacteriaceae counts (EBC), and Escherichia coli O157 prevalence . The treatment reduced the prevalence of E . coli O157 on hides from 56% to 34% and the prevalence on preevisceration carcasses from 23% to 3% . The treatment decreased aerobic plate counts from 4.9 log CFU/100 cm2 to 3.4 log CFU/100 cm2 and EBC from 3.1 log CFU/100 cm2 to 2.0 log CFU/100 cm2 on preevisceration carcasses . The treatment of hides did not result in any detectable CPC contamination of the chilled carcasses . These data indicated that a 1% CPC treatment preceded by a water wash was capable of reducing hide prevalence of E . coli O157 from as high as 80% to less than 50%, resulting in preevisceration carcass prevalence of 5% or less . We conclude that water washing followed by an antimicrobial treatment, such as CPC, has great potential as an effective hide intervention step and should be further evaluated for implementation as a processing step after stunning and before hide removal. Biochem Biophys Res Commun, 2004 May 7, 317(3), 851 - 6 Resistance to imipenem, cefepime, and cefpirome associated with mutation in Omp36 osmoporin of Enterobacter aerogenes; Thiolas A et al.; Enterobacter aerogenes develops increased multidrug resistance via a functional alteration of outer-membrane permeability associated with a decrease in porin function . We have sequenced the gene coding the major porin of Enterobacter aerogenes, omp36 . The sequence shows a high similarity with the Klebsiella pneumoniae ompK36 gene and is closely related to the enterobacterial OmpC family . Sequence analysis of several Omp36 issued from clinical strains indicated variability in putative cell-surface exposed domains . Interestingly, substitution Gly112Asp was observed in the conserved eyelet L3 region of the porin produced by two strains, C and 3 . This substitution is associated with a high general beta-lactam resistance observed in these isolates and with alteration of pore properties previously described in strain 3 porin {Mol . Microbiol . 41 (2001) 189} . This is the first genetic identification of impermeability-mediated resistance to beta-lactams in various clinical E . aerogenes strains. Mil Med, 2004 Mar, 169(3), 194 - 7 Management of postirradiation infection: lessons learned from animal models; Brook I et al.; Ionizing radiation depresses host defenses and enhances susceptibility to local and systemic infection due to endogenous or exogenous microorganisms . Exposure of mice to a lethal dose of ionizing 60Co-gamma radiation induces a dose-related reduction in the number of both aerobic and anaerobic bacteria from 10(10-12) to 10(4-6) per gram of stool within 4 days . The number of anaerobic bacteria stays low, but the number of Enterobacteriaceae per gram of stool increases significantly up to 10(9) by the 12th day after irradiation . This increase is associated with bacterial translocation of these organisms and fatal bacteremia . The use of quinolones in the irradiated animals was effective in controlling systemic endogenous Gram-negative infection after irradiation . Supplementation with penicillin prevented treatment failures due to Streptococcus spp . and increased survival . Quinolones given for 21 days also were effective in management of systemic exogenous infections due to orally ingested Klebsiella pneumoniae and Pseudomonas aeruginosa . Effectiveness of quinolones may be attributed to inhibition of exogenous organism growth within the gut lumen while preserving the anaerobic gut flora as well as their systemic antibacterial activity . Based on these findings, antimicrobial agents recommended for therapy of infection after exposure to irradiation are: ciprofloxacin, levofloxacin, ceftriaxone, cefepime, gentamicin +/- amoxicillin, or vancomycin. Emerg Infect Dis, 2004 Jan, 10(1), 94 - 9 Fluoroquinolones protective against cephalosporin resistance in gram-negative nosocomial pathogens; Schwaber MJ et al.; In a matched case-control study, we studied the effect of prior receipt of fluoroquinolones on isolation of three third-generation cephalosporin-resistant gram-negative nosocomial pathogens . Two hundred eighty-two cases with a third-generation cephalosporin-resistant pathogen (203 with Enterobacter spp., 50 with Pseudomonas aeruginosa, and 29 with Klebsiella pneumoniae) were matched on length of stay to controls in a 1:2 ratio . Case-patients and controls were similar in age (mean 62 years) and sex (54% male) . Variables predicting third-generation cephalosporin resistance were surgery (p = 0.005); intensive care unit stay (p < 0.001); and receipt of a b-lactam/b-lactamase inhibitor (p < 0.001), a ureidopenicillin (p = 0.002), or a third-generation cephalosporin (p < 0.001) . Receipt of a fluoroquinolone was protective against isolation of a third-generation cephalosporin-resistant pathogen (p = 0.005) . Interventional studies are required to determine whether replacing third-generation cephalosporins with fluoroquinolones will be effective in reducing cephalosporin resistance and the effect of such interventions on fluoroquinolone resistance. Pediatr Dermatol, 2004 Mar-Apr, 21(2), 113 - 6 Microbiology of infected hemangiomas in children; Brook I; Bacterial infections are a common complication of hemangiomas in children . The objective of this study was to establish the aerobic and anaerobic microbiology of infected hemangiomas . A retrospective 8-year review of clinical and microbiology laboratory records from patients with secondarily infected hemangiomas was carried out . Specimens from infected sites were processed for the presence of aerobic and anaerobic bacteria . Bacterial growth was present in 32 of 38 specimens . Aerobic bacteria alone were recovered in 12 infected hemangiomas (37.5%), anaerobic bacteria alone in 8 (33%), and mixed aerobic and anaerobic flora in 12 (37.5%) . A total of 80 isolates (47 aerobes and 33 anaerobes) were recovered, giving an average of 2.5 isolates per specimen (1.5 aerobes and 1.0 anaerobes) . The highest number of isolates were recovered in infections of the perineum (3.7 per site) and the legs (2.8 per site) . The predominant aerobic isolates were Staphylococcus aureus, group A beta-hemolytic streptococci, and Enterobacteriaceae . The predominant anaerobes were Peptostreptococcus sp., gram-negative bacilli, and Fusobacterium sp . Organisms that belong to the mucous membranes close to the lesions predominated in infections next to those membranes . The polymicrobial etiology of secondarily infected hemangiomas and the association of bacterial flora with the anatomic site of the lesions is thereby demonstrated. J Chin Med Assoc, 2004 Jan, 67(1), 15 - 20 Neonatal bacteremia in a neonatal intensive care unit: analysis of causative organisms and antimicrobial susceptibility; Lee NC et al.; BACKGROUND: Infections cause significant mortality and morbidity in neonates, especially the premature ones . Although there are various antibiotics can be used to combat neonatal infection, resistant strains have subsequently emerged . In an epidemiological survey, we analyzed bacterial isolates and their antibiotic susceptibilities for cases of bacteremia in a neonatal intensive care unit (NICU) of a teaching hospital . METHODS: From November 1999 to October 2001, 623 neonates admitted to the NICU were enrolled . The incidence and incidence density of bacteremia, morbidity and mortality of sepsis, as well as antibiotic susceptibility, were investigated . RESULTS: Totally, 754 blood cultures were done on 623 patients . Fifty-eight patients experienced 85 episodes of bacteremia, with 87 isolates cultured . The incidence of bacteremia in our NICU was 9.31% (58/623) with an incidence density of 10.98/1000 patient-days . The overall mortality rate was 7.22% . The case fatality rate of bacteremia was 20.7% (12/58) . The bacterial pathogens encountered, in order of frequency, were coagulase-negative Staphylococcus (29%), Staphylococcus aureus (22%), and Enterobacter cloacae (17%) . All of the gram-positive bacteria were susceptible to vancomycin, while the gram-negative bacteria were susceptible to imipenem, amikacin and ciprofloxacin . Oxacillin-resistant S . epidermidis, oxacillin-resistant S . aureus, and multi-drug resistant enterobacteriae were the leading microorganisms causing bacteremia in our NICU . CONCLUSIONS: It is an endless struggle to combat neonatal infection . Periodic evaluation of bacterial antibiotic susceptibility is necessary . More judicious selection of antibiotics and rotating antibiotic regimens should be kept in mind to reduce the resurgence of multidrug resistant strains. J Physiol Pharmacol, 2003 Dec, 54 Suppl 4, 165 - 82 Role of COX inhibition in pathogenesis of NSAID-induced small intestinal damage; Takeuchi K et al.; Nonsteroidal antiinflammatory drugs (NSAIDs) such as indomethacin decrease mucosal PGE(2) production by inhibiting cyclooxygenase (COX) activity and produce damage in the small intestine . The development of intestinal lesions as induced by indomethacin was accompanied by increases in intestinal motility, enterobacterial invasion, and myeloperoxidase (MPO) as well as inducible nitric oxide synthase (iNOS) activity, together with the up-regulation of COX-2 and iNOS mRNA expression . Neither the selective COX-1 inhibitor, SC-560, nor the selective COX-2 inhibitor, rofecoxib, alone caused intestinal damage, but their combined administration produced lesions . SC-560, but not rofecoxib, caused intestinal hypermotility, bacterial invasion and the expression of COX-2 as well as iNOS mRNAs, yet the iNOS and MPO activity was increased only when rofecoxib was administered together with SC-560 . Although SC-560 inhibited the PG production, the level of PGE(2) was recovered, in a rofecoxib-dependent manner . The intestinal hypermotility response to indomethacin was prevented by both 16,16-dimethyl PGE(2) and atropine but not ampicillin, yet all these agents inhibited not only the bacterial invasion but also the expression of COX-2 as well as the iNOS activity in the intestinal mucosa following indomethacin treatment, resulting in preventing the intestinal lesions . These results suggest that inhibition of COX-1, despite causing intestinal hypermotility, bacterial invasion and iNOS expression, up-regulates the expression of COX-2, and the PGE(2) derived from COX-2 counteracts deleterious events caused by COX-1 inhibition and maintains the mucosal integrity . These sequences of events explain why intestinal damage occurs when both COX-1 and COX-2 are inhibited. Arq Bras Cardiol, 2004 Mar, 82(3), 287 - 90 Epub 2004 Apr 05. {Vascular prosthesis infection in thoracic aorta surgery: review of the experience and a case report illustrating treatment with an unconventional technique}; Fontes RD et al.; We report the case of a 37-year-old-female patient who had undergone a Bentall procedure at our service and returned with intense chest pain and acute aortic dissection type III, which was diagnosed and clinically treated . One year after this episode, this dissection expanded, and the patient underwent surgery with interposition of a Dacron graft in the descending aorta . In the immediate postoperative period, the patient experienced left bronchopneumonia and was discharged afebrile and in good condition . One month after discharge, she returned with fever and toxemia . Pleural empyema was diagnosed, and she underwent an exploratory thoracotomy that did not confirm this diagnosis, but revealed intense effusion thickening . Four months after the exploratory thoracotomy, Klebsiella pneumoniae and Enterobacter sp were isolated in a blood culture . Magnetic resonance imaging revealed shapes compatible with perigraft infection . With this clinical and laboratory picture, graft removal was indicated as was axillo-bifemoral grafting . Surgery was successfully performed, the patient was discharged in good condition, and remains well after a 57-month follow-up without complications . The methods used for diagnosis and treatment of prosthesis infection in thoracic aorta surgery are discussed. J Clin Microbiol, 2004 Apr, 42(4), 1542 - 6 Evaluation of the automated phoenix system for potential routine use in the clinical microbiology laboratory; Donay JL et al.; A comparative study was designed to evaluate the identification (ID) and antimicrobial susceptibility testing (AST) performances of the BD Phoenix Automated Microbiology System (Becton Dickinson Diagnostic Systems {BD}, Pont de Claix, France) . A total of 305 single clinical isolates were collected, and comparisons were made with routine manual methods in use in our microbiology laboratories . The percentages of correct IDs were 93.3, 89.4, 91.8, and 85.7% for enterobacteria, nonfermenting gram-negative bacilli, staphylococci, and streptococci-enterococci, respectively . The median ID time was 3 h, and the median time for AST was 10 h 30 min . AST results showed variable percentages of errors for the different antibiotics . None of the enterobacteria and 0.3% of Pseudomonas aeruginosa isolates showed a very major error (VME) . Only one strain of Staphylococcus aureus showed a VME with oxacillin . We demonstrate here the efficiency of the Phoenix system, which can be used for the majority of strains encountered in a university-based laboratory, for ID and AST. Infect Control Hosp Epidemiol, 2004 Mar, 25(3), 226 - 30 Nosocomial pediatric bacteremia: the role of intravenous set contamination in developing countries; Macias AE et al.; OBJECTIVE: To assess the rate of bacterial contamination of intravenous administration sets at their rubber injection ports and matching infusates . DESIGN: Cultures of injection ports and infusate during 26 visits to 4 hospitals . SETTING: Four public general pediatric hospitals in Mexico City with substandard care practices . PATIENTS: Hospitalized pediatric patients receiving intravenous solutions . RESULTS: Overall, 176 of 251 injection ports were contaminated (70.1%; 95% confidence interval {CI95}, 64.5% to 75.8%), 35 (13.9%; CI95, 9.7% to 18.2%) with gram-negative rods, primarily of the tribe Klebsielleae . Cultures of infusates were positive in 17 cases (6.8%, CI95, 3.7% to 9.9%), 5 of which grew gram-negative rods (2%; CI95, 0.6% to 4.6%) . In 3 cases (1.2%), the same species with gram-negative rods was found in the infusates and on the injection ports . During one visit, 8 clustered cases of injection port contamination with a clonal Enterobacter cloacae were found; this agent was also found in the blood culture, intravenous fluid, and parenteral nutrition of one patient . Inadequate chlorination of tap water, a potential risk factor, was recorded during 22 visits (84.6%) . CONCLUSION: These data suggest that external contamination of the intravenous administration set could play a role in infusate contamination. Infect Control Hosp Epidemiol, 2004 Mar, 25(3), 192 - 7 Dynamics of bacterial hand contamination during routine neonatal care; Pessoa-Silva CL et al.; OBJECTIVE: To evaluate the dynamics of bacterial contamination of healthcare workers' (HCWs) hands during neonatal care . SETTING: The 20-bed neonatal unit of a large acute care teaching hospital in Geneva, Switzerland . METHODS: Structured observation sessions were conducted . A sequence of care began when the HCW performed hand hygiene and ended when the activity changed or hand hygiene was performed again . Alcohol-based handrub was the standard procedure for hand hygiene . An imprint of the five fingertips of the dominant hand was obtained before and after hand hygiene and at the end of a sequence of care . Regression methods were used to model the final bacterial count according to the type and duration of care and the use of gloves . RESULTS: One hundred forty-nine sequences of care were observed . Commensal skin flora comprised 72.4% of all culture-positive specimens (n = 360) . Other microorganisms identified were Enterobacteriaceae (n = 55, 13.8%); Staphylococcus aureus (n = 10, 2.5%); and fungi (n = 7, 1.8%) . Skin contact, respiratory care, and diaper change were independently associated with an increased bacterial count; the use of gloves did not fully protect HCWs' hands from bacterial contamination . CONCLUSIONS: These data confirm that hands become progressively contaminated with commensal flora and potential pathogens during neonatal care, and identify activities at higher risk for hand contamination . They also reinforce the need for hand hygiene after a sequence of care, before starting a different task, and after glove removal. J Microbiol Immunol Infect, 2004 Feb, 37(1), 35 - 8 Characteristics of nosocomial bacterial meningitis in children; Lin PC et al.; Nosocomial meningitis is uncommon in children . We reviewed the medical records of all children who developed bacterial meningitis at least 72 hours after admission to Mackay Memorial Hospital for the period July 1992 through June 2000 . Clinical manifestations, predisposing factors, pathogens, and outcomes were analyzed . Twenty-two cases of nosocomial meningitis were identified, comprising 9.2% (22/239) of all pediatric cases of bacterial meningitis during the study period . The male-to-female ratio was 14:8 . All patients were younger than 6 months of age except for one, who was 7 years old . The mean duration between admission and onset of meningitis was 15.3 days (range, 3 to 58 days) . Twenty-two organisms were isolated, including 13 Gram-negative bacteria (59%) and 9 Gram-positive bacteria (41%) . The most common pathogen was Escherichia coli (5 cases), followed by Enterobacter cloacae (3), Staphylococcus aureus (3), and Chryseobacterium meningosepticum (3) . Seventeen patients (77%) had concomitant bacteremia . Predisposing factors for acquisition of nosocomial meningitis included previous treatment with broad-spectrum antibiotics (68%), prematurity with very low birth weight (41%), and total parenteral nutrition (32%) . Two patients (9%) had previous neurosurgical intervention . Four patients (18%) died, 3 of whom were low birth weight premature infants . Nine patients (41%) had sequelae, including developmental delay, hydrocephalus, hearing impairment, and epilepsy . Neurosurgery was not a significant risk factor for the development of nosocomial meningitis, while very low birth weight played an important role . Previous intraventricular hemorrhage or hydrocephalus, prematurity with very low birth weight, infection with Gram-negative bacteria, and prior broad-spectrum antibiotic administration were associated with poor outcome. J Microbiol Immunol Infect, 2004 Feb, 37(1), 1 - 7 Effects of PNPG on cell growth cycle, motility machinery and quorum sensing in Serratia marcescens; Wei JR et al.; p-Nitrophenylglycerol (PNPG) effectively inhibits swarming of the enterobacterium Proteus mirabilis . The underlying mechanism of inhibition is unclear . We have now found that both PNPG also inhibits motility and swarming in another enterobacterium, Serratia marcescens . While the peak promoter activities of the flagellar master operon (flhDCSm), the flagellin structural gene (hagSm) and the nuclease gene (nucASm) in S . marcescens increased with increasing PNPG concentration, the expression of these genes was delayed in accordance with the reduced growth rate . As the quorum-sensing system is involved in the regulation of swarming in S . marcescens, we also examined the effect of PNPG on the production of quorum-sensing signal molecules and found that their expression was delayed with a reduced level . PNPG, therefore, had a pleiotropic effect on all aspects of S . marcescens physiology relating to swarming . The underlying molecular mechanism remains to be elucidated. Spinal Cord, 2004 Apr, 42(4), 230 - 4 Bacteriological investigation of infected pressure ulcers in spinal cord-injured patients and impact on antibiotic therapy; Heym B et al.; STUDY DESIGN: Retrospective . OBJECTIVES: To improve the use of bacteriological results for treating spinal cord-injured patients with infected pressure ulcers . SETTING: Microbiology and Orthopaedics Department, Ambroise Pare University Hospital, Boulogne-Billancourt, France . METHODS: Tissue specimens, sampled at the end of the surgical intervention from unbridled and cleaned ulcers were analysed . Drainage liquids were cultured at day 1 (D1) and day 5 (D5) postsurgery . For part of the patients, a presurgery superficial sample was analysed and compared with the surgical and postsurgical samples . RESULTS: In all, 168 surgical samples from 101 patients, 183 D1 and 104 D5 wound drainage liquids were included in this study . Out of the 168 surgical samples 17 (10%) had a negative culture, whereas 151 (90%) had a positive culture . For drainage liquids, the culture was negative in 48% and 56% of the samples at D1 and D5, respectively . The most frequently isolated species were enterobacteria, followed by staphylococci and streptococci . CONCLUSION: Culturing deep tissue specimens sampled from the surgically cleaned and unbridled ulcers allows for the isolation of the bacterial species that are really involved in the ulcer infection . As the identification of these bacteria and their antibiotic susceptibility are available, when the culture results of the D1 postsurgical drainage liquid is also available, it is easier to choose targeted antibiotic treatment. Zhonghua Nei Ke Za Zhi, 2004 Feb, 43(2), 112 - 6 {The impact of gastric colonization on the pathogenesis of ventilator-associated pneumonia}; Li HY et al.; OBJECTIVE: To investigate the risk factors for gastric bacterial colonization and its role in the endogenous pathogenesis of ventilator-associated pneumonia (VAP) . METHODS: The type and concentration of gastric colonized bacteria and its relationship with the time when samples were collected, and with the type and occurrence order of the pathogens detected in samples from lower respiratory tract after the onset of VAP were analyzed dynamically in the patients with tracheal-intubation or tracheotomy in intensive care unit (ICU) . RESULTS: (1) The isolation rate of colonized bacteria in gastric cavity was associated with the pH of gastric juice . When the pH of gastric juice was > 4, the isolation rate of Gram-negative bacilli (GNB) in gastric cavity markedly increased, achieving 52.5% in VAP group, and the incidence of VAP was higher (P = 0.017) . The pH value of gastric juice was positively correlated with the logarithmic concentration of GNB in gastric cavity (P = 0.001) . (2) As the duration of intubation prolonged, the isolation rate of enterobacteriaceae in VAP group increased, which was 45.2% on the fifth day of intubation . In contrast, the isolation rate in non-VAP group was 11.1% (P < 0.01) . (3) The colonization of enterobacteriaceae in gastric cavity was 1 - 2 days earlier than that in oropharynx . The order was statistically significant (P = 0.015) . (4) The reverse order of stomach-pharynx-lower respiratory tract colonization was found in 12 cases of the total 52 VAP patients and the order of stomach to lower respiratory tract colonization was found in 3 cases . CONCLUSIONS: (1) The pH value of gastric juice proved to be the major factor which influenced the colonization of bacteria especially GNB in gastric cavity . (2) The gastric cavity was probably a colonization place of GNB especially enterobacteriaceae . (3) The enterobateriaceae in gastric cavity tended to colonize reversely to oropharynx . (4) The phenotypic analysis of the pathogens showed that the reverse stomach-pharynx-lower respiratory tract infection route existed in VAP patients. Zhonghua Gan Zang Bing Za Zhi, 2004 Mar, 12(3), 167 - 9 {Dynamic variability of intestinal flora and endotoxin in rat with fulminate hepatic failure}; Li LJ et al.; OBJECTIVE: To investigate the dynamic variability of intestinal flora and endotoxins in rats with fulminate hepatic failure . METHODS: Establishing the fulminate hepatic failure models by intraperitoneal injection of Galactosamine . Forty Sprague-Dawley rats were divided into three groups: group A (n=10) were killed at the beginning of the experiment as control; while Group B (n=12) and C (n=18), the fulminate hepatic failure models, were killed 24 and 48 hours respectively after successful induction . Then, the contents of the jejunum, ileum and colon descendents were collected and a quantitative analysis was made about intestinal flora . Meanwhile, the concentrations of endotoxin in portal vein and right ventricle were determined and so were those in contents of ileums and colons . RESULTS: Our experiments showed that the livers of rats in group B were injured most seriously among three groups, and a minor recovery of hepatic function was observed in group C with the decrease of total bile acids (P< 0.05) . Analysis on intestinal flora show: the intestinal enterobacteriacea increase and the lactobacillus decrease in group B (P< 0.01 in jejunum and ileum and P<0.05 in colon) . The comparisons between group C and B showed that the enterobacteriacea in the former decreased in both jejunum and colon (P< 0.05) while the number of lactobacillus recovered in the jejunum of group C (P<0.05) . Quantitative analysis on endotoxins showed that the ileum endotoxin increased in group B (P< 0.05) and in group C, endotoxins in ileum and colons also increased (vs . control, P<0.01); portal endotoxin in group B showed higher level than that in group A and C (P< 0.01) . CONCLUSION: The alteration of intestinal flora was observed in fulminate hepatic failure rats . Abnormal intestinal flora might lead to incline of endotoxin in ileum, colon and portal vein, while the recovery of normal intestinal flora would decrease the level of portal endotoxin. J Pak Med Assoc, 2004 Jan, 54(1), 20 - 4 Intraprostatic tissue infection in catheterised patients in comparison to controls; Talpur AN et al.; OBJECTIVE: To determine the effect of indwelling urinary catheter on frequency of intraprostatic tissue infection and posto-perative morbidity in patients with benign prostatic hyperplasia undergoing transurethral resection of prostate (TURP) . METHODS: Frequency of intraprostatic tissue and urinary infection, prevalent organisms, histopathology of prostatic tissue, post-operative morbidity were analyzed for 25 consecutive patients' of clinically diagnosed benign prostatic hyperplasia who underwent TURP in catheterized and non-catheterized groups . RESULTS: Patients mean age was 62.2+7.9 years . In non-catheterized group nocturia and frequency were the most common symptoms . Ninety two percent and 28% patients acquired intraprostatic tissue infection in catheterized and non-catheterized group respectively, while 80% of catheterized patients and 24% of the non-catheterized patients had bacteriuria . Catheterized patients had E . coli as prevalent organism both in intaprostatic tissue and urine (34.8% and 40% respectively) . E . coli Serratia and enterobacter were equally prevalent in intraprostatic tissue of non-catheterized patients . Enterobacter was the prevalent organism in urine (50%) of these patients . There was no significant difference in the presence of non-specific inflammatory cells in the two groups . Catheterized group showed significantly high frequency of fever >38.5 degrees C and hematuria for more than 24 hours . CONCLUSION: Catheterization significantly increases the frequency of intraprostatic tissue infection as well as morbidity of TURP. Curr Microbiol, 2004 Feb, 48(2), 124 - 9 Decolorization of azo dyes with Enterobacter agglomerans immobilized in different supports by using fluidized bed bioreactor; Moutaouakkil A et al.; Immobilized cells of Enterobacter agglomerans, able to reduce azo dyes enzymatically, were used as a biocatalyst for the decolorization of synthetic medium containing the toxic azo dye methyl red (MR) . This bacterial strain exhibits high ability to completely decolorize 100 mg/L of MR after only 6 h of incubation under aerobic conditions . Cells of E . agglomerans were immobilized in calcium alginate, polyacylamide, cooper beech, and vermiculite, and were used for the decolorization of MR from synthetic water by using a fluidized bed bioreactor . The highest specific decolorization rate was obtained when E . agglomerans was entrapped in calcium alginate beads and was of about 3.04 mg MR/g cell/h with a 50% conversion time ( t(1/2)) of about 1.6 h . Moreover, immobilized cells in calcium alginate continuously decolorized MR even after seven repeated experiments without significant loss of activity, while polyacrylamide-, cooper beech-, and vermiculite-immobilized cells retained only 62, 15, and 13% of their original activity, respectively. Curr Microbiol, 2004 Mar, 48(3), 167 - 74 Cloning and characterization of the gene encoding for OMP-PD porin: the major Photobacterium damsela outer membrane protein; Gribun A et al.; The outer membrane protein of Photobacterium damsela (OMP-PD) and the gene encoding for this porin protein were isolated and characterized . The deduced amino acid sequence of the OMP-PD monomer has 338 amino acids and a calculated molecular weight of 36,951 Da . This sequence includes a 22-amino acid signal peptide at the N-terminal, which is not found when the monomer is located in the outer membrane . Native OMP-PD protein forms a trimeric structure of approximately 110 kDa . It exhibits resistance to proteases, and it can be cleaved only following denaturation by SDS . The degree of identity of the OMP-PD amino acid sequence to porins from the Enterobacteriaceae was only 24% . Identity to Vibrio or Photobacterium porins was 38% and 48%, respectively . Nevertheless, the multiple alignment of this sequence with other structurally defined Enterobacteria porins demonstrated that the location of the 16 beta-strands and eight external loops, including a larger external L3 loop, are conserved in OMP-PD . These results, together with the previously known ability of OMP-PD to form an ion channel in artificial liposomes, strongly support its role as a porin in P . damsela and will help further investigations into the role of OMP-PD in P . damsela pathogenicity. Roum Arch Microbiol Immunol, 2002 Oct-Dec, 61(4), 285 - 91 Resistance pattern of extended-spectrum beta-lactamase producing Enterobacteriaceae isolates; Tuchilus C et al.; Gram-negative pathogens harboring extended-spectrum beta-lactamases (ESBL) are becoming an increasing therapeutic problem in many wards . The aim of our work was to study ESBL production by Enterobacteriaceae strains from Eastern Romania and their antimicrobial resistance . We selected 54 clinical isolates among 1068 enterobacteria according to their susceptibility spectrum (National Committee for Clinical Laboratory Standards, 1999) . Antimicrobial susceptibility tests were performed using the Rapid ATB E gallery of mini API system (BioMerieux) and by a macrodilution method in Mueller-Hinton agar following standard procedure of the National Committee for Clinical Laboratory Standards (NCCLS) . ESBL production was established by using both double disk synergy test (DDT) and Expert computer program of mini API . The isoelectric point (pI) was determined by isoelectric focusing in polyacrylamide gel and revealed by nitrocefin . As references we used beta-lactamases with known pI . The Expert computer program of mini API confirms the positive DDT test for all selected strains . Almost all strains displayed resistance to ampicillin, ampicillin/sulbactam or third generation cephalosporins and aztreonam . By IEF we identified 51 strains which have a unique enzyme . IEF pattern showed presence of two enzymes in three Escherichia coli strains . According to our results, the ESBL TEM-type are the most common for the studied isolates . The production of extended-spectrum beta-lactamases and the presence of the multiresistant of antimicrobial agents reflect, probably, the over use of third generation cephalosporins in Eastern Romania. Arch Immunol Ther Exp (Warsz), 2004 Jan-Feb, 52(1), 43 - 9 Serological characterization of the O-specific polysaccharide of Providencia alcalifaciens O23; Torzewska A et al.; INTRODUCTION: The genus Providencia belongs to the Enterobacteriaceae family and currently consists of five species: P . alcalifaciens, P . heimbachae, P . rettgerii, P . rustigianii and P . stuartii . The serological classification scheme of P . alcalifaciens, P . rustigianii and P . stuartii includes 63 O-serogroups and 30 H-serogroups . The O-antigenic specificity is defined by the structure of the O-antigen (O-specific polysaccharide--OPS), a part of the lipopolysaccharide (LPS, endotoxin), one of the major components of the outer membrane of gram-negative bacteria and an important virulence factor of these bacteria . Among the bacteria of the Enterobacteriaceae family, the genus Providencia is one of the least studied in respect to its LPS structure and antigenic specificity . Studies of the chemical structures and the serological specificity of the O-antigens aim at the elucidation of the molecular basis of the serological classification of Providencia sp . MATERIALS AND METHODS: LPS and alkali-treated LPS of P . alcalifaciens O23 and serologically related P . rustigianii O14, P . mirabilis O13 and P . myxofaciens as well as O-antiserum against P . alcalifaciens O23 were used . Serological characterization of P . alcalifaciens O23 O-specific polysaccharide was done by use enzyme immunosorbent assay (EIA), passive hemolysis test (PHT) as well as by inhibition and sodium deoxycholate polyacrylamide gel electrophoresis (DOC-PAGE) of LPS and Western blot . RESULTS AND CONCLUSIONS: The OPS of P . alcalifaciens, O23, contains an N-(D-glucuronoyl)-N-{(R)-1-carboxyethyl}-L-lysine residue (GlcAAlaLys) . The LPS of P . alcalifaciens, O23, and other LPSs containing AlaLys from Providencia and Proteus strains were tested with rabbit anti-P . alcalifiaciens O23 serum . The serological data showed that a GlcAAlaLys-associated epitope plays a role as an antigenic determinant in the P . alcalifaciens O23 OPS and revealed the particular importance of glucuronic acid and the carboxyethyl group for the binding of O23-specific antibodies. Poult Sci, 2004 Mar, 83(3), 384 - 91 Salmonella in commercially manufactured feeds; Jones FT et al.; We collected 886 samples (68 feed ingredient samples, 189 dust samples, and 629 feed samples) from 3 feed mills each of which produced between 100,000 and 400,000 tons of feed a year . Samples were collected on 3 d (Monday, Wednesday, and Friday), during 2 seasons (early spring and summer), and between 0700 and 1700 h approximately once per hour . Samples were collected from 5 locations within each mill: ingredient receiving, at the mixer, at the pellet mill, from pellet coolers, and at load-out . Temperatures were taken of the samples obtained at the pellet mill immediately following collection . All samples were analyzed for Enterobacteriaceae counts (EC) and Salmonella . The data confirm that feed ingredients and dust can be a major source of Salmonella contamination in feed mills . There were no differences (P < 0.05) in the Salmonella contamination rates of samples collected in spring as compared with samples collected in summer . Salmonella contamination rates were observed to be higher in samples collected on Friday compared with samples collected on Monday or Wednesday, an effect that may be management related . Data collected at the pellet mill clearly illustrate the uneven distribution of Salmonella contamination in feed as well as the need for control of dust around the pellet mill . Feed samples (both mash and pellets) contaminated with Salmonella contained significantly higher EC than samples not contaminated with Salmonella . Thus, EC may provide some indication of the likelihood of Salmonella contamination in feed samples. Otolaryngol Pol, 2003, 57(6), 813 - 7 {Long term administration of itraconazole with surgical treatment in fungal and bacterial infections of the paranasal sinuses}; Jordan J et al.; A case of chronic paranasal sinuses with recurrent polyposis caused by miscellaneous infection--fungal (Aspergillus, Candida) and bacterial (Staphylococcus aureus, Enterobacter, Streptococcus) is described . The patient underwent 5 times surgical treatment (polypectomies, sinus operations) . Good result was achieved after 2-years application of itraconazole and local Amphotericin B. Rev Soc Bras Med Trop, 2003 Nov-Dec, 36(6), 711 - 7 {Spontaneous bacterial peritonitis}; Strauss E et al.; Spontaneous bacterial peritonitis occurs in 30% of patients with ascites due to cirrhosis leading to high morbidity and mortality rates . The pathogenesis of spontaneous bacterial peritonitis is related to altered host defenses observed in end-stage liver disease, overgrowth of microorganisms, and bacterial translocation from the intestinal lumen to mesenteric lymph nodes . Clinical manifestations vary from severe to slight or absent, demanding analysis of the ascitic fluid . The diagnosis is confirmed by a number of neutrophils over 250/mm3 associated or not to bacterial growth in culture of an ascites sample . Enterobacteriae prevail and Escherichia coli has been the most frequent bacterium reported . Mortality rates decreased markedly in the last two decades due to early diagnosis and prompt antibiotic treatment . Third generation intravenous cephalosporins are effective in 70% to 95% of the cases . Recurrence of spontaneous bacterial peritonitis is common and can be prevented by the continuous use of oral norfloxacin . The development of bacterial resistance demands the search for new options in the prophylaxis of spontaneous bacterial peritonitis; probiotics are a promising new approach, but deserve further evaluation . Short-term antibiotic prophylaxis is recommended for patients with cirrhosis and ascites shortly after an acute episode of gastrointestinal bleeding. Mem Inst Oswaldo Cruz, 2003 Dec, 98(8), 1093 - 5 Epub 2004 Mar 09. Detection of pathogenic bacteria in skin lesions of patients with chiclero's ulcer . Reluctant response to antimonial treatment; Isaac-Marquez AP et al.; We investigated the bacterial flora present in skin lesions of patients with chiclero's ulcer from the Yucatan peninsula of Mexico using conventional culture methods (11 patients), and an immunocolorimetric detection of pathogenic Streptococcus pyogenes (15 patients) . Prevalence of bacteria isolated by culture methods was 90.9% (10/11) . We cultured, from chiclero's ulcers (60%), pathogenic bacterial such as Staphylococcus aureus (20%), S . pyogenes (1.6%), Pseudomonas aeruginosa (1.6%), Morganella morganii (1.6%), and opportunist pathogenic bacteria such as Klebsiella spp . (20.0%), Enterobacter spp . (20%), and Enterococcus spp . (20%) . We also cultured coagulase-negative staphylococci in 40% (4/10) of the remaining patients . Micrococcus spp . and coagulase-negative staphylococci constituted the bacterial genuses more frequently isolated in the normal skin of patients with chiclero's ulcer and healthy individuals used as controls . We also undertook another study to find out the presence of S . pyogenes by an immunocolorimetric assay . This study indicated that 60% (9/15) of the ulcerated lesions, but not normal controls, were contaminated with S . pyogenes . Importantly, individuals with purulent secretion and holding concomitant infections with S . pyogenes, S . aureus, P . aeruginosa, M . morganii, and E . durans took longer to heal Leishmania (L.) mexicana infections treated with antimonial drugs . Our results suggest the need to eliminate bacterial purulent infections, by antibiotic treatment, before starting antimonial administration to patients with chiclero's ulcer. Antimicrob Agents Chemother, 2004 Apr, 48(4), 1384 - 96 In vitro antimicrobial activity of doripenem, a new carbapenem; Ge Y et al.; The doripenem MICs at which 90% of the tested strains were inhibited ranged from 0.03 to 1 microg/ml for 10 species of Enterobacteriaceae (n = 351), from 0.03 to 0.12 microg/ml for oxacillin-susceptible staphylococci (n = 119), from 4 to 32 microg/ml for oxacillin-resistant staphylococci (n = 64), from < or =0.008 to 0.06 microg/ml for penicillin-susceptible streptococci (n = 132), and from 1 to 4 microg/ml for penicillin-resistant streptococci (n = 51) . Overall, doripenem demonstrated in vitro activity similar to that of meropenem against gram-negative pathogens and to that of imipenem against gram-positive pathogens. Antimicrob Agents Chemother, 2004 Apr, 48(4), 1249 - 55 Dissemination of CTX-M-type beta-lactamases among clinical isolates of Enterobacteriaceae in Paris, France; Eckert C et al.; We analyzed 19 clinical isolates of the family Enterobacteriaceae (16 Escherichia coli isolates and 3 Klebsiella pneumoniae isolates) collected from four different hospitals in Paris, France, from 2000 to 2002 . These strains had a particular extended-spectrum cephalosporin resistance profile characterized by a higher level of resistance to cefotaxime and aztreonam than to ceftazidime . The bla(CTX-M) genes encoding these beta-lactamases were involved in this resistance, with a predominance of bla(CTX-M-15) . Ten of the 19 isolates produced both TEM-1- and CTX-M-type enzymes . One strain (E . coli TN13) expressed CMY-2, TEM-1, and CTX-M-14 . bla(CTX-M) genes were found on large plasmids . In 15 cases the same insertion sequence, ISEcp1, was located upstream of the 5' end of the bla(CTX-M) gene . In one case we identified an insertion sequence designated IS26 . Examination of the other three bla(CTX-M) genes by cloning, sequencing, and PCR analysis revealed the presence of a complex sul1-type integron that includes open reading frame ORF513, which carries the bla gene and the surrounding DNA . Five isolates had the same plasmid DNA fingerprint, suggesting clonal dissemination of CTX-M-15-producing strains in the Paris area. Ann Clin Microbiol Antimicrob . 2004 Mar 25;3(1):3. Prevalence of antimicrobial resistance in bacteria isolated from central nervous system specimens as reported by U.S . hospital laboratories from 2000 to 2002; Jones ME et al.; BACKGROUND: Bacterial infections of the central nervous system, especially acute infections such as bacterial meningitis require immediate, invariably empiric antibiotic therapy . The widespread emergence of resistance among bacterial species is a cause for concern . Current antibacterial susceptibility data among central nervous system (CNS) pathogens is important to define current prevalence of resistance . METHODS: Antimicrobial susceptibility of pathogens isolated from CNS specimens was analyzed using The Surveillance Database (TSN(R)) USA Database which gathers routine antibiotic susceptibility data from >300 US hospital laboratories . A total of 6029 organisms derived from CNS specimen sources during 2000-2002, were isolated and susceptibility tested . RESULTS: Staphylococcus aureus (23.7%) and Streptococcus pneumoniae (11.0%) were the most common gram-positive pathogens . Gram-negative species comprised approximately 25% of isolates . The modal patient age was 1 or <1 year for most organisms . Prevalence of MRSA among S . aureus from cerebrospinal fluid (CSF) and brain abscesses were 29.9-32.9% . Penicillin resistance rates were 16.6% for S . pneumoniae, 5.3% for viridans group streptococci, and 0% for S . agalactiae . For CSF isolates, ceftriaxone resistance was S . pneumoniae (3.5%), E . coli (0.6%), Klebsiella pneumoniae (2.8%), Serratia marcescens (5.6%), Enterobacter cloacae (25.0%), Haemophilus influenzae (0%) . Listeria monocytogenes and N . meningitidis are not routinely susceptibility tested . CONCLUSIONS: Resistance is commonly detected, albeit still at relatively low levels for key drugs classes such as third-generation cephalosporins . This data demonstrates the need to consider predominant resistance phenotypes when choosing empiric therapies to treat CNS infections. Biochem J, 2004 Jul 15, 381(Pt 2), 527 - 36 Combinational clustering of receptors following stimulation by bacterial products determines lipopolysaccharide responses; Triantafilou M et al.; The innate immune system has the capacity to recognize a wide range of pathogens based on conserved PAMPs (pathogen-associated molecular patterns) . In the case of bacterial LPS (lipopolysaccharide) recognition, the best studied PAMP, it has been shown that the innate immune system employs at least three cell-surface receptors: CD14, TLR4 (Toll-like receptor 4) and MD-2 protein . CD14 binds LPS from Enterobacteriaceae and then transfers it to MD-2, leading to TLR4 aggregation and signal transduction . LPS analogues such as lipid IVa seem to act as LPS antagonists in human cells, but exhibit LPS mimetic activity in mouse cells . Although TLR4 has been shown to be involved in this species-specific discrimination, the mechanism by which this is achieved has not been elucidated . The questions that remain are how the innate immune system can discriminate between LPS from different bacteria as well as different LPS analogues, and whether or not the structure of LPS affects its interaction with the C