Eur J Biochem, 2002 Nov, 269(22), 5712 - 21 Purification and catalytic properties of a CO-oxidizing:H2-evolving enzyme complex from Carboxydothermus hydrogenoformans; Soboh B et al.; From the membrane fraction of the Gram-positive bacterium Carboxydothermus hydrogenoformans, an enzyme complex catalyzing the conversion of CO to CO2 and H2 was purified . The enzyme complex showed maximal CO-oxidizing:H2-evolving enzyme activity with 5% CO in the headspace (450 U per mg protein) . Higher CO concentrations inhibited the hydrogenase present in the enzyme complex . For maximal activity, the enzyme complex had to be activated by either CO or strong reductants . The enzyme complex also catalyzed the CO- or H2-dependent reduction of methylviologen at 5900 and 180 U per mg protein, respectively . The complex was found to be composed of six hydrophilic and two hydrophobic polypeptides . The amino-terminal sequences of the six hydrophilic subunits were determined allowing the identification of the encoding genes in the preliminary genome sequence of C . hydrogenoformans . From the sequence analysis it was deduced that the enzyme complex is formed by a Ni-containing carbon monoxide dehydrogenase (CooS), an electron transfer protein containing four {4Fe-4S} clusters (CooF) and a membrane bound {NiFe} hydrogenase composed of four hydrophilic subunits and two membrane integral subunits . The hydrogenase part of the complex shows high sequence similarity to members of a small group of {NiFe} hydrogenases with sequence similarity to energy conserving NADH:quinone oxidoreductases . The data support a model in which the enzyme complex is composed of two catalytic sites, a CO-oxidizing site and a H2-forming site, which are connected via a different iron-sulfur cluster containing electron transfer subunits . The exergonic redox reaction catalyzed by the enzyme complex in vivo has to be coupled to energy conservation, most likely via the generation of a proton motive force. Schweiz Rundsch Med Prax, 2002 Oct 9, 91(41), 1691 - 8 {Whipple disease--a rare systemic disease}; Dancygier H et al.; Whipple's disease is a rare, without antibiotic treatment deadly systemic infectious disease caused by the ubiquitary Gram positive bacterium Tropheryma whipplei . The agent can be demonstrated by light and electron microscopy and by PCR in tissues and body fluids affected . Men are affected more often than women . Most patients lose weight, have diarrhea, abdominal pain and arthralgias . In 10-40% of cases involvement of the CNS, often asymptomatic, is observed . PAS-positive macrophages in the lamina propria of duodenal biopsies are typical but not pathognomonic of Whipple's disease . Nowadays, the infectious agent should also be demonstrated by PCR . Treatment, mostly as a sequential therapy, is with antibiotics that cross the blood-brain barrier and should last for at least 1 year in order to avoid relapses . Before ending therapy the absence of Tropheryma whipplei in duodenal tissue and cerebrospinal fluid must be confirmed. Xenobiotica, 2002 Oct, 32(10), 907 - 24 Pharmacokinetics, toxicokinetics, distribution, metabolism and excretion of linezolid in mouse, rat and dog; Slatter JG et al.; 1 . Linezolid (ZYVOX), the first of a new class of antibiotics, the oxazolidinones, is approved for treatment of Gram-positive bacterial infections . 2 . The aim was to determine the absorption, distribution, metabolism and excretion (ADME) of linezolid in mouse, rat and dog in support of preclinical safety studies and clinical development . 3 . Conventional replicate study designs were employed in animal experiments, and biofluids were assayed by HPLC or HPLC-MS . 4 . Linezolid was rapidly absorbed after p.o . dosing with an p.o . bioavailability of > 95% in rat and dog, and > 70% in mouse . Twenty-eight-day i.v./p.o . toxicokinetic studies in rat (20-200mg kg(-1) day(-1)) and dog (10-80 mg kg(-1) day(-1)) revealed neither a meaningful increase in clearance nor accumulation upon multiple dosing . 5 . Linezolid had limited protein binding (<35%) and was very well distributed to most extravascular sites, with a volume of distribution at steady-state (V(ss)) approximately equal to total body water . 6 . Linezolid circulated mainly as parent drug and was excreted mainly as parent drug and two inactive carboxylic acids, PNU-142586 and PNU-142300 . Minor secondary metabolites were also characterized . In all species, the clearance rate was determined by metabolism . 7 . Radioactivity recovery was essentially complete within 24-48 h . Renal excretion of parent drug and metabolites was a major elimination route . Parent drug underwent renal tubular reabsorption, significantly slowing parent drug excretion and allowing a slow metabolic process to become rate-limiting in overall clearance . 8 . It is concluded that ADME data were relatively consistent across species and supported the rat and dog as the principal non-clinical safety species. Appl Environ Microbiol, 2002 Nov, 68(11), 5480 - 7 Expression of duplicate msa genes in the salmonid pathogen Renibacterium salmoninarum; Rhodes LD et al.; Renibacterium salmoninarum is a gram-positive bacterium responsible for bacterial kidney disease of salmon and trout . R . salmoninarum has two identical copies of the gene encoding major soluble antigen (MSA), an immunodominant, extracellular protein . To determine whether one or both copies of msa are expressed, reporter plasmids encoding a fusion of MSA and green fluorescent protein controlled by 0.6 kb of promoter region from msa1 or msa2 were constructed and introduced into R . salmoninarum . Single copies of the reporter plasmids integrated into the chromosome by homologous recombination . Expression of mRNA and protein from the integrated plasmids was detected, and transformed cells were fluorescent, demonstrating that both msa1 and msa2 are expressed under in vitro conditions . This is the first report of successful transformation and homologous recombination in R . salmoninarum. Gene, 2002 Sep 18, 298(1), 41 - 8 Molecular and functional analysis of the Rickettsia typhi groESL operon; Radulovic S et al.; The groESL operon from an obligate, intracellular, Gram-negative bacterium Rickettsia typhi, the etiologic agent of murine typhus, was cloned and sequenced . The sequence analysis of 2229 bp of the groESL operon reveals two open reading frames of 288 nucleotides (groES) and 1653 nucleotides (groEL) separated by 20 nucleotides . The deduced amino acid sequence of R . typhi GroES and GroEL shows a high degree of identity with other bacterial GroES and GroEL . Reverse transcriptase-polymerase chain reaction and Northern blot analysis indicated that both groES and groEL are transcribed as a single mRNA . The transcriptional start point at 81 nucleotides upstream of the groES start codon was determined by primer extension . The promoter analysis shows no regulatory CIRCE element as it is known for many Gram-positive and Gram-negative bacteria . However, it contains the sequence similar to the putative sigma(70)-dependent promoter and lacks the -35 sequence of the putative sigma(32)-dependent promoter . Complementation assay by R . typhi groESL in a temperature sensitive Escherichia coli groEL mutant restored significant growth ability at non-permissive temperature. Res Microbiol, 2002 Sep, 153(7), 405 - 15 Phylogeny of phosphoryl transfer proteins of the phosphoenolpyruvate-dependent sugar-transporting phosphotransferase system; Hu KY et al.; Some bacteria lack sugar permeases of the bacterial phosphotransferase system (PTS) but encode within their genomes phosphoryl transfer proteins of the PTS that probably function in regulation . These proteins include homologues of HPr (PtsH), the ATP-dependent HPr(ser) kinase/phosphatase (PtsK) and the PEP-dependent HPr(his) kinase known as Enzyme I (PtsI) . We identify all currently sequenced homologues of these proteins, multiply align their sequences and construct phylogenetic trees in order to derive functional, structural and evolutionary conclusions . We show that no bacterium possesses more than one HPr kinase and that these proteins are probably all orthologous . alpha-Proteobacteria possess truncated HPr kinases which probably serve a unified regulatory function together with other PTS proteins . The Enzymes I are orthologous in all Gram-positive bacteria and some Gram-negative bacteria, but other Gram-negative bacteria exhibit paralogues that fall into 5 functional types . No bacterium with a fully sequenced genome exhibits all of these types . With the exception of the classical Enzymes I, each of these functional types exhibits a distinctive set of accompanying domains, usually with a characteristic domain order . One functional type, the fructose-specific type, includes two phylogenetically different subgroups with different domain orders . The results establish that domain associations occurred early during evolutionary history of the PTS, and that subsequent domain rearrangements occurred rarely . Our findings define the evolutionary histories of these important bacterial proteins and provide guides for functional assignment of PTS-related proteins encoded by genes revealed by genome sequencing. J Small Anim Pract, 2002 Oct, 43(10), 456 - 8 Bronchogenic cyst in a German shepherd dog; Dahl K et al.; Intermittent episodes of fever, severe dyspnoea and cyanosis were observed in a two-and-a-half-year-old male German shepherd dog . The clinical signs were of sudden onset but disappeared within eight hours, in five almost identical episodes over a period of 40 days . Radiological examination of the thorax revealed a large rounded area of radiolucency surrounded by a thin radiopaque structure . The lesion increased in volume between examinations, but otherwise remained similar in appearance . It could not be confirmed from the radiographs whether the structure was situated in lung tissue or mediastinum . Bronchoscopic examination revealed no abnormal findings and there was no bacterial growth from samples taken 30 days after the onset of symptoms . A large, vesicular structure, partly adherent to the mediastinum, was surgically removed from the left cranial lung lobe . The structure was 8 to 10 cm in diameter and partly filled with mucoid pink fluid . Histological examination revealed that the wall contained traces of bronchial epithelial cells . On bacteriological examination of the fluid, Gram-positive, mucus-forming bacteria were present . The definitive gross pathological and histopathological diagnosis was a bronchogenic cyst which had developed secondarily to bronchiectasis . Bronchitis and peribronchitis were also identified . The dog recovered well after surgery and, at the time of writing, was in active duty as a police dog, showing no signs of respiratory illness. J Bacteriol, 2002 Nov, 184(22), 6270 - 9 Helicobacter pylori expresses an autolytic enzyme: gene identification, cloning, and theoretical protein structure; Marsich E et al.; Helicobacter pylori is an important pathogen of the gastric system . The clinical outcome of infection is thought to be correlated with some genetic features of the bacterium . However, due to the extreme genetic variability of this organism, it is hard to draw definitive conclusions concerning its virulence factors . Here we describe a novel H . pylori gene which expresses an autolytic enzyme that is also capable of degrading the cell walls of both gram-positive and gram-negative bacteria . We designated this gene lys . We found this gene and observed its expression in a number of unrelated clinical strains, a fact that suggests that it is well conserved in the species . A comparison of the nucleotide sequences of lys and the hypothetical gene HP0339 from H . pylori strain ATCC 26695 revealed almost total identity, except for the presence of an insertion consisting of 24 nucleotides in the lys sequence . The coding sequences of lys and HP0339 show a high degree of homology with the coding sequence of bacteriophage T4 lysozyme . Because of this similarity, it was possible to model the three-dimensional structures of both the lys and HP0339 products. Immunobiology, 2002 Sep, 205(4-5), 575 - 94 Pulmonary innate immune proteins and receptors that interact with gram-positive bacterial ligands; Palaniyar N et al.; The two major gram-positive bacterial (GPB) ligands are peptidoglycan (PGN) and lipoteichoic acid (LTA) . These polymeric LTA and highly organized PGN contain repeating carbohydrate moieties, which are potential targets for pattern recognition molecules . The major pattern recognition proteins and receptors, which bind GPB, either have a lectin, PGN recognition, collagen or leucine-rich repeat (LRR) domain . The soluble innate immune proteins (IIPs) that bind to PGN and LTA include pulmonary collectins surfactant-associated proteins (SP-) A and D, lectin-like pentraxins C-reactive protein (CRP) and serum amyloid P component (SAP), and sCD14 . Membrane-anchored lectin or lectin-like group members include macrophage mannose receptor (MR), complement receptor 3 (CR3, or Mac-1, or integrin CD11b/CD18), scavenger receptor A (SRCL-1), lectin-like oxidized LDL receptor 1 (LOX-1), and GPI-anchored CD14 . Although Toll-like receptor (TLR) 2 and 4, and CD14 contain extracellular LRR domains, only TLRs have a cytoplasmic domain for signal transduction . Three of the four recently discovered human PGN recognition proteins (PGRP) have a transmembrane domain, and hence, considered as true receptors for GPB . Since lysozyme is the only known pulmonary enzyme that can lyse bacterial cell wall PGN, other innate immune molecules appear to be responsible for signalling and enhancing the clearance of GPB infection from the lung . Interestingly, pulmonary collectins bind not only to GPB ligands but also to the receptors, CD14 and TLR, and antigen processing cells such as dentritic cells . These complex interactions appear to play major roles in linking innate and adaptive immunity, and maintaining a pathogen-free lung with minimal, or no inflammation. Blood, 2003 Jan 1, 101(1), 71 - 7 Epub 2002 Aug 08. High-dose cyclophosphamide with autologous lymphocyte-depleted peripheral blood stem cell (PBSC) support for treatment of refractory chronic autoimmune thrombocytopenia; Huhn RD et al.; Patients with refractory chronic autoimmune thrombocytopenia (AITP) have a significant risk of morbidity and mortality related to hemorrhage . High-dose (HD) cytotoxic therapy may produce remissions but entails risks related to myelosuppression . Hematopoietic stem cell support with lymphocyte-depleted grafts may accelerate hematologic recovery and concomitantly reduce repopulation by autoreactive immunocytes . Fourteen patients with chronic AITP, in whom multiple prior therapies including corticosteroids, splenectomy, intravenous immunoglobulin, and various cytotoxic or immunomodulatory regimens had failed, were treated with HD cyclophosphamide (50 mg/kg/d) and autologous granulocyte colony-stimulating factor (G-CSF)-mobilized leukocytes depleted of lymphocytes by immunomagnetic CD34(+) selection . There were no significant adverse events related to G-CSF, intravenous device insertion, or leukapheresis . Treatment-related complications included transient hemorrhagic cystitis (1 patient), vaginal bleeding (2 patients), gastrointestinal bleeding (1 patient), epistaxis (1 patient), and antibiotic-responsive febrile neutropenia (all patients) . The mean time to absolute neutrophil count (ANC) more than 500/mm(3) was 9 +/- 0.6 days . Eight patients experienced antibiotic-responsive gram-positive bacteremia . A median of 2 platelet transfusions was required for stem cell mobilization, intravenous catheter insertion, and apheresis and a median of 9 platelet transfusions was required during hematopoietic recovery . Six patients obtained durable complete responses (platelet counts > 100 000/mm(3) without other therapy) with maximum follow-up of 42 months . Two additional patients obtained durable partial responses (platelet counts significantly increased over baseline with reduced medication requirements and cessation of bleeding complications) . This therapeutic approach is feasible for patients with severe chronic AITP, a substantial proportion of whom may obtain durable remissions . Larger controlled trials are recommended. Glycoconj J, 2001 Sep, 18(9), 715 - 22 Signaling networks controlling mucin production in response to Gram-positive and Gram-negative bacteria; McNamara N et al.; Human lung cells exposed to pathogenic bacteria upregulate the production of mucin, the major macromolecular component of mucus . Generally this upregulation is beneficial for the host, however, in the lungs of cystic fibrosis patients, overproduction of mucin can lead to the plugging of pulmonary airways . Mucus plugging impedes airflow and creates an environment that is highly compartmentalized: those bacteria within the mucus layer are shielded from high doses of antibiotics whereas those outside the mucus are exposed . These conditions augment mutation rate and the development of drug resistance in bacteria that colonize the lungs of cystic fibrosis patients . While therapeutic inhibition of mucin induction would improve airflow and reduce antibiotic resistance in these patients, the challenge is to develop drugs that block excessive mucin production while leaving beneficial aspects of the response intact . To do this, we must understand the molecular mechanisms underlying mucin production . Here we review the signal transduction pathways that control mucin production in response to Gram-positive and Gram-negative bacteria. Infection, 2002 Oct, 30(5), 317 - 9 Vertebral osteomyelitis and endocarditis of a pacemaker lead due to Granulicatella (Abiotrophia) adiacens; Rosenthal O et al.; Systemic infection due to Granulicatella (formerly Abiotrophia), a species of nutrition-deficient gram-positive cocci, is rare . We present the case of a 68-year-old diabetic male who presented with back pain and a history of fever and chills . Imaging studies revealed vertebral osteomyelitis of the Th 10/11 region . Transesophageal echocardiography disclosed a vegetation adjacent to the pacemaker lead and blood cultures grew Granulicatella adiacens . A diagnosis of vertebral osteomyelitis and endocarditis due to G . adiacens was made and the patient improved with bed rest and medical treatment alone . Granulicatella ssp . should always be part of the differential diagnosis of fastidious bacteria in vertebral osteomyelitis and endocarditis. Infect Immun, 2002 Nov, 70(11), 6129 - 39 Contributions of the N- and C-terminal domains of surfactant protein d to the binding, aggregation, and phagocytic uptake of bacteria; Hartshorn KL et al.; Collectins play important roles in host defense against infectious microorganisms . We now demonstrate that the serum collectins mannose-binding lectin (MBL) and conglutinin have less ability to bind to, aggregate, and enhance neutrophil uptake of several strains of gram-negative and gram-positive bacteria than pulmonary surfactant protein D (SP-D) . Collectins are composed of four major structural domains (i.e., N-terminal, collagen, and neck and carbohydrate recognition domains) . To determine which domains of SP-D are responsible for its greater bacterial binding or aggregating activity, activities of chimeric collectins containing the N-terminal and collagen domains of SP-D coupled to the neck recognition domains and carbohydrate recognition domains (CRD) of MBL or conglutinin (SP-D/Cong(neck+CRD) and SP-D/MBL(neck+CRD)) were tested . The SP-D/Cong(neck+CRD) and SP-D/MBL(neck+CRD) chimeras bound to and aggregated the bacteria more strongly than did wild-type MBL or conglutinin . SP-D/MBL(neck+CRD) also enhanced neutrophil uptake of bacteria more so than MBL . Hence, the SP-D N-terminal and/or collagen domains contribute to the enhanced bacterial binding and aggregating activities of SP-D . In prior studies, SP-D/Cong(neck+CRD) and SP-D/MBL(neck+CRD) had increased ability to bind to influenza virus compared not only with that of conglutinin or MBL but with that of wild-type SP-D as well . In contrast, the chimeras had either reduced or unchanged ability to bind to or aggregate bacteria compared to that of wild-type SP-D . Hence, although replacement of the neck recognition domains and CRDs of SP-D with those of MBL and conglutinin conferred increased viral binding activity, it did not favorably affect bacterial binding activity, suggesting that requirements for optimal collectin binding to influenza virus and bacteria differ. Trends Microbiol, 2002 Oct, 10(10), 452 - 5 Why are rod-shaped bacteria rod shaped? Koch AL. Generally speaking, bacteria grow and divide indefinitely, and as long as the growth conditions are maintained they retain constant dimensions and shapes with little variation . How they do this is a question that I have been considering for three decades . Here, I discuss two hypothetical mechanisms, one for Gram-positive rods and the other for Gram-negative rods . These mechanisms are consistent with what is known, but make some unproven assumptions. Inorg Chem, 2002 Oct 21, 41(21), 5475 - 8 Corynebactin and a serine trilactone based analogue: chirality and molecular modeling of ferric complexes; Bluhm ME et al.; Because the hydrolysis of ferric ion makes it very insoluble in aerobic, near neutral pH environments, most species of bacteria produce siderophores to acquire iron, an essential nutrient . The chirality of the ferric siderophore complex plays an important role in cell recognition, uptake, and utilization . Corynebactin, isolated from Gram-positive bacteria, is structurally similar to enterobactin, a well-known siderophore first isolated from Gram-negative bacteria, but contains L-threonine instead of L-serine in the trilactone backbone . Corynebactin also contains a glycine spacer unit in each of the chelating arms . A hybrid analogue (serine-corynebactin) has been prepared which has the trilactone ring of enterobactin and the glycine spacer of corynebactin . The chirality and relative conformational stability of the three ferric complexes of enterobactin, corynebactin, and the hybrid have been investigated by molecular modeling (including MM3 and pBP86/DN density functional theory calculations) and circular dichroism spectra . While enterobactin forms a Delta-ferric complex, corynebactin is Lambda . The hybrid serine-corynebactin forms a nearly racemic mixture, with the Lambda-conformer in slight excess . Each ferric complex has four possible isomers depending on the metal chirality and the conformation of the trilactone ring . For corynebactin, the energy difference between the two possible Lambda conformations is 2.3 kcal/mol . In contrast, only 1.5 kcal/mol separates the inverted Lambda- and normal Delta-configuration for serine-corynebactin . The small energy difference of the two lowest energy configurations is the likely cause for the racemic mixture found in the CD spectra . Both the addition of a glycine spacer and methylation of the trilactone ring (serine to threonine) favor the Lambda-conformation . These structural changes suffice to change the chirality from all Delta (enterobactin) to all Lambda (corynebactin) . The single change (glycine spacer) of the hybrid ferric serine-corynebactin gives a mixture of Delta and Lambda, with the Lambda in slight excess. Asian J Surg, 2002 Jul, 25(3), 209 - 13 Rapid diagnosis of fungal infection in patients with acute necrotizing pancreatitis by polymerase chain reaction; Zhang WZ et al.; OBJECTIVE: This study was conducted to assess the rapid diagnosis of fungal infections in patients with acute necrotizing pancreatitis by polymerase chain reaction {PCR} using universal primers targeting the 18S rRNA gene . METHODS: In this study, a PCR assay was developed to identify clinically isolated fungi, and both PCR technique and conventional culture were used to detect fungi in 37 samples from patients with acute necrotizing pancreatitis . RESULTS: A 197-bp fragment was amplified by PCR from all the clinically isolated fungal strains . This fragment was not isolated from gram-positive, gram-negative bacteria or human leucocytes . Thirty-seven samples of necrotic tissue or peripancreatic fluid from 11 patients were also analyzed, and eight samples were positive for fungi by PCR, six of which were also positive by conventional culture . The whole PCR procedure was completed within 7 hours . CONCLUSION: PCR can be used to diagnose fungal infection secondary to acute necrotizing pancreatitis rapidly and sensitively. Microbes Infect, 2002 Sep, 4(11), 1149 - 56 Iron acquisition by Gram-positive bacterial pathogens; Brown JS et al.; For the majority of bacterial pathogens, acquisition of iron from host proteins is a prerequisite for growth during infection . The mechanisms by which Gram-negative bacteria obtain iron from host proteins have been well described, but only recently has substantial progress been made in identifying these mechanisms for Gram-positive bacterial pathogens . This review provides an overview of the existing knowledge on the genetic basis of iron transport for important Gram-positive pathogens. Int J Syst Evol Microbiol, 2002 Sep, 52(Pt 5), 1825 - 9 Nocardia pseudovaccinii sp . nov; Kim KK et al.; Comparative 16S rDNA studies of Nocardia type and reference strains revealed that strain DSM 43406T, identified as Nocardia vaccinii, was wrongly classified . The strain was aerobic, gram-positive and produced scarce, white, branched aerial mycelium and a beige-red substrate mycelium . The reverse side of the colonies was yellow-orange . It showed chemotaxonomic markers that were consistent with its classification in the genus Nocardia . The mycolic acids had chain lengths from 50 to 58 carbon atoms . The 16S rDNA sequence showed the highest similarity to Nocardia nova (97.7%) and N . vaccinii (97.6%), but the strain could be clearly separated from these species and other members of the N . vaccinii cluster by significant differences in biochemical test results and unique fatty acid and mycolic acid patterns . These data led to the conclusion that the isolate represents a novel species within the genus Nocardia, for which the name Nocardia pseudovaccinii sp . nov . is proposed . The type strain is strain AR 368,38366-20T (= DSM 43406T = NRRL B-24154T). Int J Syst Evol Microbiol, 2002 Sep, 52(Pt 5), 1681 - 5 Alicyclobacillus acidiphilus sp . nov., a novel thermo-acidophilic, omega-alicyclic fatty acid-containing bacterium isolated from acidic beverages; Matsubara H et al.; A novel thermo-acidophilic bacterium was isolated from an acidic beverage that had the odour of guaiacol . The cells are aerobic, gram-positive, spore-forming rods . The organism, strain TA-67T, grows at temperatures from 20 to 55 degrees C (optimum, 50 degrees C) and at pH values from 2.5 to 5.5 (optimum, pH 3.0) . It possesses omega-cyclohexane fatty acid as a major cellular fatty acid . The G+C content of the DNA is 54.1 mol% . Phylogenetic analysis of the 16S rRNA gene sequences indicated that strain TA-67T constituted a distinct lineage in the Alicyclobacillus cluster, with Alicyclobacillus acidoterrestris as the closest neighbour (96.6% homology) . Phenotypically, it is similar to, but can be distinguished from, omega-cyclohexane fatty acid-possessing alicyclobacilIi (A . acidoterrestris, Alicyclobacillus acidocaldarius, Alicyclobacillus hesperidum and 'Alicyclobacillus mali') by the morphology of spores and sporangia, by the growth response to different temperatures, and by the profiles for acid production from carbon sources . It is the alicyclobacillus that produces guaiacol, a causative substance for an 'off' flavour of orange juice . On the basis of the phenotypic and phylogenetic evidence, it is concluded that strain TA-67T represents a new species of the genus Alicyclobacillus, for which the name Alicyclobacillus acidiphilus is proposed . The type strain is TA-67T (= DSM 14558T = IAM 14935T = NRIC 6496T). Int J Syst Evol Microbiol, 2002 Sep, 52(Pt 5), 1609 - 14 Kytococcus schroeteri sp . nov., a novel Gram-positive actinobacterium isolated from a human clinical source; Becker K et al.; A strain of a gram-positive, coccoid, yellow-pigmented bacterium was isolated from human blood . The bacterium was aerobic, non-encapsulated and non-motile . Phenotypically, the bacterium closely resembled Kytococcus sedentarius, but could be distinguished from this species by physiological tests and chemotaxonomic investigations . The peptidoglycan type is L-Lys-Glu2, variation A4alpha . The predominant menaquinones are MK-8 and MK-7 . The major cellular fatty acids are iso-C17:1, iso-C17:0, iso-C15:0 and anteiso-C17:0 . The strain contains catalase and does not produce acid from carbohydrates . The ability to hydrolyse Tween 80 and the lack of alpha-glucosidase activity are the most characteristic features . The results of comparative 16S rDNA analysis revealed that the strain represents a novel species within the genus Kytococcus, for which the name Kytococcus schroeteri sp . nov . is proposed . The type strain is strain Muenster 2000T (= DSM 13884T = CCM 4918T). Am J Infect Control, 2002 Oct, 30(6), 355 - 72 Hygienic hand antiseptics: should they not have activity and label claims against viruses? Sattar SA, Springthorpe VS, Tetro J, Vashon R, Keswick B. Enteric and respiratory viruses are among the most frequent causes of human infections, and hands play an important role in the spread of these and many other viral diseases . Regular and proper hand hygiene by caregivers and food handlers in particular is essential to decontaminate hands and potentially interrupt such spread . What would be considered a proper decontamination of hands? Handwashing with regular soap and water is often considered sufficient, but what of hygienic handwash and handrub antiseptic products? Are they more effective? The evidence suggests that some clearly are . Activity against bacteria may not reflect the ability of hygienic hand antiseptics to deal with viruses, especially those that are nonenveloped . In spite of the acknowledged importance of hands as vehicles for viruses, there is a lack of suitable regulatory mechanism for handwash or handrub products to make claims of efficacy against viruses . This is in contrast with the ability of general-purpose disinfectants to make antiviral claims, although transmission of viruses from surfaces other than those of reusable medical devices may play only a minor role in virus transmission . This review discusses the (1) . recent information on the relative importance of viruses as human pathogens, particularly those causing enteric and respiratory infections; (2) . the survival of relevant viruses on human hands in comparison with common gram-negative and gram-positive bacteria; (3) . the potential of hands to transfer or receive such contamination on casual contact; (4) . role of hands in the spread of viruses; (5) . the potential of hygienic measures to eliminate viruses from contaminated hands; (6) . relative merits of available protocols to assess the activity of hygienic hand antiseptics against viruses; and (7) . factors considered crucial in any tests to assess the activity of hygienic hand antiseptics against viruses . In addition, this review proposes surrogate viruses in such testing and discusses issues for additional consideration by researchers, manufacturers, end-users, and regulators. Proc Natl Acad Sci U S A, 2002 Oct 15, 99(21), 13861 - 6 Epub 2002 Oct 01. Innate recognition of bacteria by a macrophage cytosolic surveillance pathway; O'Riordan M et al.; Host recognition of bacterial pathogens is a critical component of the immune response . Intracellular bacterial pathogens are able to evade the humoral immune system by residing within the host cell . Here we show the existence of an innate host surveillance mechanism in macrophages that specifically distinguishes bacteria in the cytosol from bacteria in the vacuole . Recognition of Gram-positive and Gram-negative bacterial products by this surveillance system results in transcription of the ifnb gene . The activation of cytosol-specific signaling is associated with translocation of NF-kappaB into the nucleus and phosphorylation of the p38 mitogen-activated protein (MAP) kinase . Activation of the p38 kinase is required for the induction of gene expression by the cytosolic surveillance pathway . Our studies suggest that infection by intracellular bacterial pathogens results in an immune response distinct from that of infection by extracellular bacterial pathogens. FEMS Microbiol Lett, 2002 Sep 10, 214(2), 183 - 8 Purification and some properties of phospho-beta-galactosidase from the Gram-negative oral bacterium Leptotrichia buccalis ATCC 14201; Thompson J; Phospho-beta-galactosidase (P-beta-gal; EC 3.2.1.85) is induced during growth of Leptotrichia buccalis ATCC 14201 on lactose and lactulose . The enzyme has been purified to electrophoretic homogeneity (M(r) approximately 53 kDa, pI approximately 4.8), and kinetic parameters have been determined using the chromogenic analog o-nitrophenyl-beta-D-galactopyranoside-6-phosphate as substrate . Both ATP and galactose-6-phosphate are inhibitors of P-beta-gal activity . Microsequence analysis has identified the first 32 residues from the N-terminus of the protein, and by comparative sequence alignment the enzyme can be assigned to Family 1 of the glycosylhydrolase superfamily . Polyclonal antibody against the enzyme permits the highly specific immuno-detection of P-beta-gal in cell-free extracts of L . buccalis . Although described previously in several Gram-positive species, this is the first reported purification of P-beta-gal from a Gram-negative organism. Nihon Kokyuki Gakkai Zasshi, 2002 Jun, 40(6), 525 - 9 {A case of pulmonary squamous cell carcinoma coexisting with pulmonary actinomycosis}; Nagaoka T et al.; A 71-year-old man was referred to our hospital complaining of cough . Chest radiography revealed a mass opacity in the right upper lung field . A transbronchial biopsy specimen revealed non-specific inflammatory changes . Percutaneous lung aspiration biopsy under ultrasound guidance demonstrated gram-positive rods, suggesting actinomyces . On the diagnosis of pulmonary actinomycosis, the patient was treated with penicillin-G and his symptoms were relieved . In a three-month follow-up, the mass shadow in the right upper lung field was found to have increased in size . Squamous cell lung cancer was diagnosed on the basis of repeated transbronchial tumor biopsies, and right upper lobectomy was performed . Most cases of pulmonary actinomycosis have been diagnosed from post-surgical tumor specimens taken on suspicion of the presence of lung cancer . However, the lung cancer in this case was difficult to diagnose because the lung cancer was co-existent with pulmonary actinomycosis. J Med Assoc Thai, 2002 Jun, 85(6), 739 - 41 Actinomycotic meningitis: report of a case; Chotmongkol V et al.; A 73-year-old man who presented with acute fever, drowsiness and confusion was reported . Two weeks prior to admission, he attended the Outpatient Department with symptoms of fever and headache for 2 weeks . Eosiophilic meningitis was initially diagnosed, which, in fact, was lymphocytic CSF pleocytosis . He was treated with a high dose of prednisolone . His symptoms improved for 1 week, then he experienced symptoms of fever and headache again . On admission, he had stiffness of the neck . Lumbar puncture showed purulent CSF with gram-positive branching filamentous organisms . CSF grew Actinomyces israelii . The patient died from brain herniation. Drug Resist Updat, 2002 Jul-Aug, 5(3-4), 119 - 25 New developments in tetracycline antibiotics: glycylcyclines and tetracycline efflux pump inhibitors; Chopra I; The tetracyclines, discovered in the 1940s, are a well-established class of antibiotics that still have a role in treating microbial infections in man . However, the widespread emergence of bacterial resistance due to efflux and ribosomal protection mechanisms has severely limited their effectiveness . A new generation of tetracyclines, the glycylcyclines, has been developed to overcome resistance to earlier tetracyclines . One of the new glycylcyclines, 9-t-butylglyclamido-minocycline (GAR-936, tigecycline) is currently undergoing clinical trials . This review considers the current status of glycylcyclines and the possibility that resistance to these agents might arise in the future . Other approaches are also being taken to address the emergence of resistance to tetracyclines . Recently, a number of tetracycline efflux pump inhibitors have been discovered that might be used in combination with earlier tetracyclines to restore their activity against resistant organisms . However, the development of tetracycline efflux pump inhibitors is complicated by the occurrence of several efflux pump sub-families and by the presence of both efflux and ribosomal protection mechanisms in the same organism, especially in naturally occurring, Gram-positive clinical isolates. J Perinat Med, 2002, 30(4), 329 - 32 Serum amyloid A protein in the early detection of late-onset bacterial sepsis in preterm infants; Arnon S et al.; In order to evaluate serum amyloid A as an early diagnostic marker of late-onset sepsis, seventy-nine preterm infants with clinically suspected sepsis and 40 healthy matched controls were assayed for serum amyloid A . In parallel, clinical and biochemical variables that are used to evaluate neonatal sepsis were compared . Forty-two episodes were diagnosed as sepsis . Serum amyloid A levels were elevated in the sepsis group (187.6 +/- 78.3 micrograms/ml), compared with infants who had no sepsis (10.2 +/- 8.3 micrograms/ml) and the control group (6.9 +/- 3.3 micrograms/ml), and were significantly higher in gram-negative compared to gram-positive sepsis (221.8 +/- 84.4 micrograms/ml vs . 48.5 +/- 22.2 micrograms/ml) . Analysis of the data suggests serum amyloid A has the highest sensitivity (100%), specificity (93%) and positive predictive value (96%) for sepsis among the clinical and biochemical parameters that were tested . In conclusion, serum amyloid A seems to be a reliable early marker for the diagnosis of late-onset sepsis in preterm infants. Plant Physiol, 1993 Dec, 103(4), 1311 - 1319 Synergistic Enhancement of the Antifungal Activity of Wheat and Barley Thionins by Radish and Oilseed Rape 2S Albumins and by Barley Trypsin Inhibitors; Terras F et al.; Although thionins and 2S albumins are generally considered as storage proteins, both classes of seed proteins are known to inhibit the growth of pathogenic fungi . We have now found that the wheat (Triticum aestivum L.) or barley (Hordeum vulgare L.) thionin concentration required for 50% inhibition of fungal growth is lowered 2- to 73-fold when combined with 2S albumins (at sub- or noninhibitory concentrations) from radish (Raphanus sativus L.) or oilseed rape (Brassica napus L.) . Furthermore, the thionin antifungal activity is synergistically enhanced (2- to 33-fold) by either the small subunit or the large subunit of the radish 2S albumins . Three other 2S albumin-like proteins, the barley trypsin inhibitor and two barley Bowman-Birk-type trypsin inhibitor isoforms, also act synergistically with the thionins (2- to 55-fold) . The synergistic activity of thionins combined with 2S albumins is restricted to filamentous fungi and to some Gram-positive bacteria, whereas Gram-negative bacteria, yeast, cultured human cells, and erythrocytes do not show an increased sensitivity to thionin/albumin combinations (relative to the sensitivity to the thionins alone) . Scanning electron microscopy and measurement of K+ leakage from fungal hyphae revealed that 2S albumins have the same mode of action as thionins, namely the permeabilization of the hyphal plasmalemma . Moreover, 2S albumins and thionins act synergistically in their ability to permeabilize fungal membranes. Perit Dial Int, 2002 May-Jun, 22(3), 345 - 9 Treatment and outcome of peritonitis in automated peritoneal dialysis, using a once-daily cefazolin-based regimen; Fielding RE et al.; OBJECTIVE: We determined the effectiveness of a once-daily cefazolin-based regimen in treating automated peritoneal dialysis (APD) peritonitis . DESIGN: We carried out a retrospective analysis of all APD peritonitis episodes treated with a once-daily cefazolin protocol . SETTING: The study was performed in a peritoneal dialysis unit in a tertiary care hospital . PATIENTS AND METHODS: We studied 60 episodes of primary peritonitis in 40 patients on APD . Each patient was treated with a vancomycin-free regimen consisting of intraperitoneal cefazolin (1.5 g IP) with gentamicin IP administered in the daytime exchange . The main outcome measures were successful treatment of peritonitis, removal of peritoneal catheter, relapse of peritonitis, and patient death . RESULTS: Gram-positive infections occurred in 35 episodes (58.3%), gram-negative Infections in 10 episodes (16.7%), culture-negative infections in 14 episodes (23.3%), and a yeast infection in 1 episode (1.7%) . Of the 60 episodes, 47 (78.3%) were successfully treated . In 10 episodes (16.7%), catheters were removed (9 for treatment failure, 1 for yeast infection) . Four patients (8%) had a relapse of infection within 4 weeks of completing antibiotic therapy . One patient (1.7%) died . CONCLUSIONS: Our results demonstrate that once-daily cefazolin with gentamicin IP is an effective treatment for APD peritonitis, with the advantage of being easy to administer and enabling patients to remain on APD during treatment. J Mol Biol, 2002 Sep 20, 322(3), 521 - 32 The TRTGn motif stabilizes the transcription initiation open complex; Voskuil MI et al.; The effect on transcription initiation by the extended -10 motif (5'-TRTG(n)-3'), positioned upstream of the -10 region, was investigated using a series of base substitution mutations in the alpha-amylase promoter (amyP) . The extended -10 motif, previously referred to as the -16 region, is found frequently in Gram-positive bacterial promoters and several extended -10 promoters from Escherichia coli . The inhibitory effects of the non-productive promoter site (amyP2), which overlaps the upstream region of amyP, were eliminated by mutagenesis of the -35 region and the TRTG motif of amyP2 . Removal by mutagenesis of the competitive effects of amyP2 resulted in a reduced dependence of amyP on the TRTG motif . In the absence of the second promoter, mutations in the TRTG motif of amyP destabilized the open complex and prevented the maintenance of open complexes at low temperatures . The open complex half-life was up to 26-fold shorter in the mutant TRTG motif promoters than in the wild-type promoter . We demonstrate that the amyP TRTG motif dramatically stabilizes the open complex intermediate during transcription initiation . Even though the open complex is less stable in the mutant promoters, the region of melted DNA is the same in the wild-type and mutant promoters . However, upon addition of the first three nucleotides, which trap RNAP (RNA polymerase) in a stable initiating complex, the melted DNA region contracts at the 5'-end in a TRTG motif promoter mutant but not at the wild-type promoter, indicating that the motif contributes to maintaining DNA-strand separation. Genome Biol . 2002 Aug 15;3(9):RESEARCH0047 . Epub 2002 Aug 15. NEAT: a domain duplicated in genes near the components of a putative Fe3+ siderophore transporter from Gram-positive pathogenic bacteria; Andrade MA et al.; BACKGROUND: Iron uptake from the host is essential for bacteria that infect animals . To find potential targets for drugs active against pathogenic bacteria, we have searched all completely sequenced genomes of pathogenic bacteria for genes relevant for iron transport . RESULTS: We identified a protein domain that appears in variable copy number in bacterial genes that are usually in the vicinity of a putative Fe3+ siderophore transporter . Accordingly, we have denoted this domain NEAT for 'near transporter' . Most of the bacterial species containing this domain are pathogenic . Sequence features indicate that the domain is anchored to the extracellular side of the membrane . The domain seems to be under high selective pressure for rapid independent duplications that are typical of sequences involved in signaling and binding . CONCLUSIONS: The NEAT domain might be functionally related to iron transport . The taxonomic specificity of this domain and its predicted extracellular position could make it an interesting target for designing new drugs against some highly pathogenic bacteria. FEBS Lett, 2002 Sep 11, 527(1-3), 193 - 8 Cupiennin 1d*: the cytolytic activity depends on the hydrophobic N-terminus and is modulated by the polar C-terminus; Kuhn-Nentwig L et al.; To investigate structural features modulating the biological activity of cupiennin 1 peptides from the spider Cupiennius salei, three truncated cupiennin 1d analogs were synthesized . The fact that their growth inhibiting effect on Gram-negative and Gram-positive bacteria, their lytic activity with human red blood cells and their insecticidal effect on Drosophila melanogaster correlates with structural properties shows that the hydrophobic N-terminal chain segment includes the major determinants of structure and activity . The polar C-terminus seems to modulate peptide accumulation at negatively charged cell surfaces via electrostatic interactions and has no important effect on the peptides' amphipathic secondary structure. J S Afr Vet Assoc, 2001 Dec, 72(4), 235 - 8 The first reported case of equine nocardioform placentitis in South Africa; Volkmann DH et al.; Since the late 1980s a distinct form of focally-extensive mucoid to mucopurulent uterine body chronic placentitis,caused by nocardioform organisms, hasbeen recognised in horses in the USA state of Kentucky and possibly in other areas . This disease has led to increasing numbers of foal losses from late abortions, still-births, prematurity, or early neonatal deaths . The foals are usually not infected, but may be small or emaciated . Modes of infection and transmission are as yet unknown . Nocardia spp . and related nocardioform bacteria as causes of equine infertility, endometritis and foal death are briefly reviewed . A case of near full-term abortion involving a Friesian mare in the Pretoria district of Gauteng Province in South Africa during February 2000, with the same placental lesion as described in the Kentucky cases, is presented . Nocardioform organisms were visualised on impression smears and histological sections of affected foetal membranes, and were also cultured . The organism has been identified at the Livestock Disease Diagnostic Center of the University of Kentucky as an Amycolatopsis sp . of the less-commonly diagnosed group of nocardioforms causing placentitis in the USA . The organism was cultured from the uterus of the mare 18 days post-foaling, but after a 2-week course of oral trimethoprim and sulphamethoxazole, based on antibiogram sensitivity testing, a uterine flush yielded no growth . A semen sample from the sire of the aborted foal did not yield any Gram-positive filamentous branching bacteria . The mare subsequently conceived to a single insemination. Adv Colloid Interface Sci, 2002 Aug 5, 98(3), 341 - 463 Analysis of different approaches for evaluation of surface energy of microbial cells by contact angle goniometry; Sharma PK et al.; Microbial adhesion on solid substrate is important in various fields of science . Mineral-microbe interactions alter the surface chemistry of the minerals and the adhesion of the bacterial cells to mineral surface is a prerequisite in several biobeneficiation processes . Apart from the surface charge and hydrophobic or hydrophilic character of the bacterial cells, the surface energy is a very important parameter influencing their adhesion on solid surfaces . There were many thermodynamic approaches in the literature to evaluate the cells surface energy . Although contact angle measurements with different liquids with known surface tension forms the basis in the calculation of the value of surface energy of solids, the results are different depending on the approach followed . In the present study, the surface energy of 140 bacterial and seven yeast cell surfaces has been studied following Fowkes, Equation of state, Geometric mean and Lifshitz-van der Waals acid-base (LW-AB) approaches . Two independent issues were addressed separately in our analysis . At first, the surface energy and the different components of the surface energy for microbial cells surface are examined . Secondly, the different approaches are evaluated for their internal consistency, similarities and dissimilarities . The Lifshitz-van der Waals component of surface energy for most of the microbial cells is realised to be approximately 40 mJ/m2 +/-10% . Equation of state and Geometric mean approaches do not possess any internal consistency and yield different results . The internal consistency of the LW-AB approach could be checked only by varying the apolar liquid and it evaluates coherent surface energy parameters by doing so . The electron-donor surface energy component remains exactly the same with the change of apolar liquid . This parameter could differentiate between the Gram-positive and Gram-negative bacterial cells . Gram-negative bacterial cells having higher electron-donor parameter had lower nitrogen, oxygen and phosphorous content on their cell surfaces . Among the four approaches, LW-AB was found to give the most consistent results . This approach provides more detailed information about the microbial cell surface and the electron-donor parameter differentiates different type of cell surfaces. Water Sci Technol, 2002, 46(1-2), 171 - 8 Role of "G-bacteria" in anaerobic substrate uptake in a SBR with no phosphorus removal; Kong YH et al.; Biomass from an SBR running with no enhanced biological phosphorus removal (EBPR) but which exhibited anaerobic assimilation of glucose and acetate, was dominated by "G-bacteria", cocci in tetrads and clusters . Extracted 16S rDNA was amplified by PCR and then analysed using Denaturing Gradient Gel Electrophoresis (DGGE) . Major bands were extracted and their sequences determined . Clone libraries were also prepared, the 16S rDNA extracted, PCR performed and the resultant fragments run by DGGE to aid in identifying the DGGE bands and provide fuller sequences than available by DGGE alone . The two approaches together allowed several bands to be identified . Probes for FISH analyses were designed for some of these in attempts to see to which phylogenetic group "G-bacteria" belonged, and whether they represented the dominant bands detected by DGGE . Then FISH/Microautoradiography (MAR) was used in attempts to see which bacteria there were assimilating substrates anaerobically . Results indicated that the "G-bacteria" were phylogenetically diverse, but mainly alpha-proteobacteria and members of the high G+C% gram-positive bacteria . Not all of these could assimilate glucose and/or acetate anaerobically, and Amaricoccus, the original "G-bacteria" of Cech and Hartman, was not detected. Kansenshogaku Zasshi, 2002 Jul, 76(7), 566 - 70 {A case of Nocardia farcinica pneumonia treated with sulfamethoxazole-trimethoprim monitoring its serum concentration}; Kachi S et al.; A 75-year-old male suffered from interstitial pneumonia in December 2000 and treated with predonisolone . The treatment was effective, and the dosage of predonisolone had been gradually tapered . In January 2001, when the dosage was 30 mg/day, he complained of cough and yellowish sputum . The chest X-ray and CT revealed bilateral infiltrations with cavities . He was treated with cefozopram and fluconazole . However, there were no improvements . The sputa of the 2nd, 3rd, 6th and 8th hospital days showed the presence of gram-positive branched rods, which were identified as Nocardia farcinica . Therefore, the treatment was changed to sulfamethoxazole-trimethoprim . During the treatment, serum concentration of sulfamethoxazole was repeatedly measured, and kept over 60 microgram/ml . He was swiftly recovered after the start of sulfamethoxazole-trimethoprim . This case was supposed to be the seventh one of N . farcinica pneumonia in Japan, and the measurement of the concentration of sulfamethoxazole was useful to determine its dosage. J Antimicrob Chemother, 2002 Sep, 50(3), 331 - 8 Sequence and genome context analysis of a new molecular class D beta-lactamase gene from Legionella pneumophila; Avison MB et al.; Legionella pneumophila Philadelphia-1 (ATCC 33152) produces a serine active site beta-lactamase . The chromosomal gene that encodes this enzyme, loxA, has been cloned by PCR using information from the L . pneumophila Philadelphia-1 genome sequencing project . LoxA is a class 2d penicillinase, and its sequence puts it into the molecular class D beta-lactamase family, although phylogenetic analysis shows that LoxA forms a distinct branch in the OXA family along with the LoxA homologue, OXA-29, from Legionella gormanii ATCC 33297(T) . Upstream of loxA on the L . pneumophila Philadelphia-1 chromosome is a two-gene locus similar to that found linked to the beta-lactamase genes of Gram-positive bacteria . The unit consists of loxI, encoding a homologue of the Gram-positive beta-lactamase expression regulator, and pbpX, encoding a putative penicillin-binding transpeptidase . Despite the presence of beta-lactamase regulator homologues, we could find no evidence of LoxA induction upon challenge of L . pneumophila Philadelphia-1 with beta-lactams. Nucleic Acids Res, 2002 Sep 1, 30(17), 3662 - 71 Predicted structure and phyletic distribution of the RNA-binding protein Hfq; Sun X et al.; Hfq, a bacterial RNA-binding protein, was recently shown to contain the Sm1 motif, a characteristic of Sm and LSm proteins that function in RNA processing events in archaea and eukaryotes . In this report, comparative structural modeling was used to predict a three-dimensional structure of the Hfq core sequence . The predicted structure aligns with most major features of the Methanobacterium thermoautotrophicum LSm protein structure . Conserved residues in Hfq are positioned at the same structural locations responsible for subunit assembly and RNA interaction in Sm proteins . A highly conserved portion of Hfq assumes a structural fold similar to the Sm2 motif of Sm proteins . The evolution of the Hfq protein was explored by conducting a BLAST search of microbial genomes followed by phylogenetic analysis . Approximately half of the 140 complete or nearly complete genomes examined contain at least one gene coding for Hfq . The presence or absence of Hfq closely followed major bacterial clades . It is absent from high-level clades and present in the ancient Thermotogales-Aquificales clade and all proteobacteria except for those that have undergone major reduction in genome size . Residues at three positions in Hfq form signatures for the beta/gamma proteobacteria, alpha proteobacteria and low GC Gram-positive bacteria groups. Appl Environ Microbiol, 2002 Sep, 68(9), 4301 - 6 Isolation of antibiotics turbomycin a and B from a metagenomic library of soil microbial DNA; Gillespie DE et al.; To access the genetic and biochemical potential of soil microorganisms by culture-independent methods, a 24,546-member library in Escherichia coli with DNA extracted directly from soil had previously been constructed (M . R . Rondon, P . R . August, A . D . Bettermann, S . F . Brady, T . H . Grossman, M . R . Liles, K . A . Loiacono, B . A . Lynch, I . A . MacNeil, M . S . Osburne, J . Clardy, J . Handelsman, and R . M . Goodman, Appl . Environ . Microbiol . 66:2541-2547, 2000) . Three clones, P57G4, P89C8, and P214D2, produced colonies with a dark brown melanin-like color . We fractionated the culture supernatant of P57G4 to identify the pigmented compound or compounds . Methanol extracts of the acid precipitate from the culture supernatant contained a red and an orange pigment . Structural analysis revealed that these were triaryl cations, designated turbomycin A and turbomycin B, respectively; both exhibited broad-spectrum antibiotic activity against gram-negative and gram-positive organisms . Mutagenesis, subcloning, and sequence analysis of the 25-kb insert in P57G4 demonstrated that a single open reading frame was necessary and sufficient to confer production of the brown, orange, and red pigments on E . coli; the predicted product of this sequence shares extensive sequence similarity with members of the 4-hydroxyphenylpyruvate dioxygenase (4HPPD) family of enzymes . Another member of the same family of genes, lly, which is required for production of the hemolytic pigment in Legionella pneumophila, also conferred production of turbomycin A and B on E . coli . We further demonstrated that turbomycin A and turbomycin B are produced from the interaction of indole, normally secreted by E . coli, with homogentisic acid synthesized by the 4HPPD gene products . The results demonstrate successful heterologous expression of DNA extracted directly from soil as a means to access previously uncharacterized small organic compounds, serving as an example of a chimeric pathway for the generation of novel chemical structures. Arch Dis Child, 2002 Sep, 87(3), 207 - 10 The emergence of resistant pneumococcal meningitis--implications for empiric therapy; McMaster P et al.; BACKGROUND: Following the emergence of penicillin and cephalosporin resistant pneumococcal meningitis in the United States, inclusion of vancomycin in empiric therapy for all suspected bacterial meningitis was recommended by the American Academy of Pediatrics . Few data are available to evaluate this policy . AIMS: To examine the management and clinical course in relation to antibiotic therapy of a large unselected cohort of children with pneumococcal meningitis in a geographic area where antibiotic resistance has recently increased . METHODS: Retrospective review of all cases of pneumococcal meningitis in a defined population (Sydney, Australia), 1994-99 . RESULTS: A total of 104 cases without predisposing illnesses were identified; timing of lumbar puncture (LP) was known in 103 . Resistance to penicillin increased from 0 to 20% over the study period . Only 57 (55%) had an early LP (prior to parenteral antibiotics); 55 (96%) had organisms on Gram stain . Severe disease (intensive care admission or death) increased significantly from 57 cases with early LP (28%) to 33 with delayed LP (42%) to 13 with no LP (62%) . Evidence of pneumococcal infection was available within 24 hours in 85% of those with delayed or no LP . Outcome was not related to empiric vancomycin use, which increased from 5% prior to 1998 to 48% in 1999 . CONCLUSION: LP is frequently delayed in pneumococcal meningitis . Based on disease severity, empiric vancomycin is most justified when LP is deferred . If an early LP is done, vancomycin can be withheld if Gram positive diplococci are not seen. Pharmacol Ther, 2002 Feb-Mar, 93(2-3), 283 - 92 Protein kinase inhibitors and antibiotic resistance; Burk DL et al.; While antibiotics revolutionized the treatment of infectious disease in the 20th century, bacterial resistance now threatens to render many of them ineffective . Aminoglycosides are a class of clinically important antibiotics used in the treatment of infections caused by Gram-positive and -negative organisms . They are bactericidal, targeting the bacterial ribosome, where they bind to the A-site and disrupt protein synthesis . Clinical resistance to these drugs occurs mainly via enzymatic inactivation by aminoglycoside-modifying enzymes that phosphorylate, adenylate, or acetylate the aminoglycoside . Those that phosphorylate (i.e., aminoglycoside kinases) have been shown to be structurally related to eukaryotic protein kinases . This was surprising, given the low degree of sequence similarity between the groups of enzymes . The nucleotide-binding site, specifically, is very similar in structure, suggesting that the two classes of enzymes share a common mechanism of phosphoryl transfer . Three strategies can be envisaged for combating aminoglycoside kinase-mediated bacterial resistance . The first involves compounds that target the antibiotic binding region . Secondly, protein kinase inhibitors have been identified that disable aminoglycoside-modifying enzymes by targeting the ATP-binding site . Lastly, compounds are being developed that exploit the bridged nature of the active site, incorporating nucleotide and substrate motifs . A strategy using bifunctional aminoglycoside dimers has also been pursued, yielding molecules that bind to the target site on the bacterial ribosome, while serving as poor substrates for modifying enzymes . This work holds out the promise that effective inhibitors of aminoglycoside-modifying enzymes may eventually restore the usefulness of aminoglycoside antibiotics. J Chromatogr A, 2002 Jun 28, 961(1), 119 - 24 Determination and pharmacokinetic study of meropenem in rat bile using on-line microdialysis and liquid chromatography; Chan YL et al.; Meropenem is a carbapenem antibiotic with a wide spectrum of activity against both Gram-positive and Gram-negative bacteria . Because of its clinical efficacy, meropenem is an excellent choice for the treatment of serious infections in both adults and children . The knowledge of tissue concentrations of antibiotic in an infection site is valuable for the prediction of treatment outcome . To investigate the biliary disposition of meropenem, we utilized a minimally invasive sampling technique with a shunt linear microdialysis probe for continuous sampling in the biliary excretion studies . Analysis of meropenem in the dialysates was achieved using a LiChrosorb RP-18 column (Merck, 250 x 4.6 mm I.D.; particle size 5 microm) maintained at ambient temperature . The mobile phase was 50 mM monosodium phosphoric acid-methanol (80:20, v/v, pH 3.0) . The UV detector wavelength was set at 298 nm . The area under the concentration-time curve and elimination half-lives of meropenem were about 6144 +/- 1494 min microg/ml and 61 +/- 17 min, respectively . This study represents a successful application of the microdialysis technique, which is an effective method for pharmacokinetic and biliary drug excretion studies. Intensive Care Med, 2002 Aug, 28(8), 1161 - 3 Epub 2002 Jul 05. The -260 C-->T promoter polymorphism of the lipopolysaccharide receptor CD14 and severe sepsis in trauma patients; Heesen M et al.; OBJECTIVE: CD14, expressed on the cell surface of monocytes and hepatic Kupffer cells, interacts with Gram-positive and Gram-negative bacteria . Upon CD14 stimulation, these cells respond with the enhanced release of cytokines involved in the pathophysiology of sepsis . The purpose of this study was to evaluate whether the genotype distribution of the -260 C-->T promoter polymorphism of the CD14 gene is associated with the development of severe sepsis in trauma patients . PATIENTS AND PARTICIPANTS: Fifty-eight severely injured blunt trauma patients with an injury severity score of 16 or more and without pre-existing chronic diseases . MEASUREMENTS AND RESULTS: Genotyping for the single nucleotide exchange polymorphism of the CD14 gene was performed by means of a real-time polymerase chain reaction with fluorescence-labeled hybridization probes . Diagnosis of severe sepsis was based on the criteria of the ACCP/SCCM criteria . Fourteen out of the 58 patients (24.6%) developed a trauma-related severe sepsis . The overall allele frequency was 0.58 for the C allele and 0.42 for the T allele . The genotype distribution (TT 0.19, CT 0.47 and CC 0.35) did not differ significantly from a previously reported control group of healthy blood donors . There was no significant difference of the genotype distribution or allele frequency between trauma patients with severe sepsis and patients with an uncomplicated clinical course . CONCLUSIONS: This pilot study suggests that the CD14 -260 polymorphism is not associated with an increased risk of severe sepsis in trauma patients. Neurosurgery, 2002 Aug, 51(2), 391 - 400; discussion 400-1 Efficacy of prophylactic antibiotic therapy in spinal surgery: a meta-analysis; Barker FG 2nd; OBJECTIVE: There is considerable variation in practice regarding the use of prophylactic antibiotic therapy in spinal operations . To date, individual studies have not demonstrated a significant benefit for prophylactic antibiotic therapy in spinal operations . METHODS: Systematic database searches for randomized prospective trials of prophylactic antibiotic therapy in spinal surgery, general neurosurgery, and orthopedic surgery were performed . Random-effects meta-analysis and Bayesian meta-regressions of treatment benefits versus baseline infection rates and other trial characteristics were performed . RESULTS: Six prospective randomized trials or trial subgroups, enrolling 843 patients, were identified, i.e., one spinal surgery trial, four general neurosurgery trials, and one general orthopedic surgery trial . No individual trial demonstrated a statistically significant effect of prophylactic antibiotic therapy for spinal surgery patients . Raw pooled infection rates were 2.2% (10 of 451 patients) with antibiotics and 5.9% (23 of 392 patients) without antibiotics . Individual trial infection rates ranged from 1.2 to 8.5% . The pooled odds ratio was 0.37 (95% confidence interval, 0.17-0.78), favoring antibiotic treatment (P < 0.01) . There was no significant heterogeneity in treatment efficacy among the trials . Bayesian meta-regression was used to test whether antibiotics were less effective in trials with low baseline infection rates and whether an optimal antibiotic regimen could be identified . There was no significant difference in antibiotic treatment effects in trials with lower baseline infection rates, in trials using antibiotics with gram-negative coverage in addition to gram-positive coverage, or in trials using multiple-dose versus single-dose regimens . Nonrandomized studies of antibiotic therapy in spinal surgery yielded greater treatment effect estimates than did randomized trials, but not significantly so . CONCLUSION: Prophylactic antibiotic therapy is beneficial for spinal surgery, even when expected infection rates without antibiotic treatment are low. Blood, 2002 Sep 1, 100(5), 1860 - 8 Role of toll-like receptor 2 (TLR2) in neutrophil activation: GM-CSF enhances TLR2 expression and TLR2-mediated interleukin 8 responses in neutrophils; Kurt-Jones EA et al.; In vitro studies as well as clinical trials indicate that the cytokines granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) enhance the ability of neutrophils (polymorphonuclear leukocytes) to eliminate microbial organisms . Toll-like receptor (TLR) proteins, homologs of the Drosophila protein Toll, have been found on the surface of mammalian cells and are important in the responses of macrophages to bacterial, viral, and fungal antigens . TLR4 is critical for the response to lipopolysaccharide (LPS) of gram-negative bacteria, while TLR2 is important for response to gram-positive bacteria, bacterial peptides, and yeast zymosan . We demonstrate that TLR2, but very little TLR4, is present on the surface of human neutrophils . In addition we demonstrate that GM-CSF and G-CSF dramatically up-regulate TLR2 and CD14 surface expression . GM-CSF treatment also up-regulates TLR2 and CD14 mRNA levels in neutrophils . In addition to increasing receptor expression, GM-CSF treatment enhanced the interleukin 8 (IL-8) secretion and superoxide priming responses of neutrophils to stimulation with TLR2 ligands, including zymosan, peptidoglycan, and lipoarabinomannan . The human monocyte response to crude bacterial LPS is composed of a TLR4-specific response to the pure LPS component and a TLR2-dependent response to associated lipopeptides . The removal of TLR2 lipopeptide components from LPS by phenol re-extraction substantially reduced both the IL-8 and superoxide response of the stimulated neutrophils, indicating that, unlike monocytes, the neutrophil response is preferentially directed to TLR2 ligands . Thus, our studies demonstrate that GM-CSF dramatically enhances the functional response of neutrophils to TLR2 ligands, including LPS-associated lipopeptides. Int J Pharm, 2002 Aug 21, 242(1-2), 271 - 5 Biodegradable implantable teicoplanin beads for the treatment of bone infections; Yenice I et al.; Parenteral antibiotic therapy for acute bone infections, soft tissue infections and osteomyelitis may result in high serum concentrations, associated with nephrotoxic, ototoxic and allergic complications . After taking these above mentioned disadvantages into consideration, recent investigations have explored the use of antibiotic-loaded biodegradable implants, incorporating antibiotics for potential use in the treatment of bone infections . In this study, biodegradable implants containing teicoplanin for the prevention or the treatment of bone infections were designed by using sodium alginate as the polymer material . Therefore, teicoplanin, a glycopeptide antibiotic, active against gram-positive bacteria was incorporated in a natural polymer in order to prepare bead formulation for implantation purpose in bone for the localized treatment of osteomyelitis . In vitro characterization was realized by determining particle size, surface characteristics, loading capacity and in vitro release characteristics of the beads. Appl Microbiol Biotechnol, 2002 Aug, 59(4-5), 419 - 25 Epub 2002 Jun 25. Protein serine/threonine kinases in signal transduction for secondary metabolism and morphogenesis in Streptomyces; Umeyama T et al.; A number of proteins in the Gram-positive bacterial genus Streptomyces are phosphorylated on their serine/threonine and tyrosine residues in response to developmental phases . AfsR is one of these proteins and acts as a transcriptional factor in both the regulation of secondary metabolism in Streptomyces coelicolor A3(2) and morphological differentiation in Streptomyces griseus . In S . coelicolor A3(2), AfsR is phosphorylated on its serine and threonine residues by more than three protein kinases whose kinase activity is enhanced by means of autophosphorylation on their serine and threonine residues . The degree of autophosphorylation of AfsK is regulated by KbpA which, by binding directly to the kinase domain of AfsK, inhibits its autophosphorylation . Phosphorylation of AfsR enhances its DNA-binding activity and causes it to bind the promoter elements, including -35, of afsS, thus resulting in activation of afsS transcription . ATPase activity of AfsR is essential for this transcriptional activation, probably because the energy available from ATP hydrolysis is required for the isomerization of the closed complex between AfsR and RNA polymerase to a transcriptionally competent open complex . afsS, encoding a 63-amino-acid protein, then activates transcription of actII-ORF4, a pathway-specific transcriptional activator in the actinorhodin biosynthetic gene cluster, in an as yet unknown way . Distribution of the afsK- afsR systems in a wide variety of Streptomyces species and the presence of many phosphorylated proteins in a given Streptomyces strain suggest that the signal transduction via not only two-component regulatory systems but also serine/threonine kinases generally regulates secondary metabolism and morphogenesis in this genus. Front Biosci, 2002 Oct 01, 7, d2045 - 57 A microbial hormone, A-factor, as a master switch for morphological differentiation and secondary metabolism in Streptomyces griseus; Horinouchi S; The Gram-positive, soil-inhabiting, filamentous bacterial genus Streptomyces employs gamma-butyrolactones as chemical signalling molecules or microbial hormones, together with their specific receptors, to regulate morphological and/or physiological differentiation . The A-factor regulatory cascade in streptomycin-producing Streptomyces griseus commences aerial mycelium formation and production of all the secondary metabolites including streptomycin . The molecular mechanism by which A-factor triggers streptomycin biosynthesis or the A-factor signal is transmitted to the streptomycin biosynthetic gene cluster has been elucidated . A transcriptional activator AdpA at one of the regulatory steps switches on many genes required for both morphological development and secondary metabolism . Most of the gene cluster for secondary metabolite biosynthesis appear to receive the A-factor signal at the respective pathway-specific transcriptional activator genes via AdpA . Accumulating evidence has shown that a pair of genes encoding a probable gamma-butyrolactone biosynthetic enzyme (AfsA-like protein) and its specific receptor (ArpA-like protein) is contained in a number of biosynthetic gene clusters for secondary metabolites in various Streptomyces species and controls the biosynthesis of the respective metabolites by activating the pathway-specific regulatory genes . Some strains contain multiple pairs of afsA-arpA . It is conceivable that, because of the almost same sites bound by various receptor proteins, the multiple ArpA-like proteins with the same or different ligand specificity in a cell competitively bind the same target sites, thus allowing the cell to grow healthy and to produce antibiotics in response to environmental conditions. Biochem J, 2002 Dec 1, 368(Pt 2), 611 - 20 Anti-microbial properties of histone H2A from skin secretions of rainbow trout, Oncorhynchus mykiss; Fernandes JM et al.; Skin exudates of rainbow trout contain a potent 13.6 kDa anti-microbial protein which, from partial internal amino acid sequencing, peptide mass fingerprinting, matrix-associated laser desorption/ionization MS and amino acid analysis, seems to be histone H2A, acetylated at the N-terminus . The protein, purified to homogeneity by ion-exchange and reversed-phase chromatography, exhibits powerful anti-bacterial activity against Gram-positive bacteria, with minimal inhibitory concentrations in the submicromolar range . Kinetic analysis revealed that at a concentration of 0.3 microM all test bacteria lose viability after 30 min incubation . Weaker activity is also displayed against the yeast Saccharomyces cerevisiae . The protein is salt-sensitive and has no haemolytic activity towards trout erythrocytes at concentrations below 0.3 microM . Reconstitution of the protein in a planar lipid bilayer strongly disturbs the membrane but does not form stable ion channels, indicating that its anti-bacterial activity is probably not due to pore-forming properties . This is the first report to show that, in addition to its classical function in the cell, histone H2A has extremely strong anti-microbial properties and could therefore help contribute to protection against bacterial invasion. Microbiol Immunol, 2002, 46(6), 409 - 12 The first isolation of Nocardia veterana from a human mycetoma; Kano R et al.; The clinical isolates from biopsy specimen human subcutaneous nodule developed orange-colored and wrinkled colonies on Sabouraud's dextrose agar at 24 C for 2 weeks . The isolates were aerobic and gram-positive . The bacteria were rod-shaped to coccoid and 1 x 5 microm in size . The assimilation tests revealed that the clinical isolate was identical to a reference strain of Nocardia veterana . A nucleotide sequence analysis of the 16S ribosomal DNA from the isolate and a reference strain of N . veterana showed 99.8% similarity . All data are consistent with the conclusion that the isolate in this human case of mycetoma is N . veterana. Aust Vet J, 2002 Jun, 80(6), 344 - 5 Pyothorax associated with a Mycoplasma sp and Arcanobacterium pyogenes in a kitten; Gulbahar MY et al.; Pyothorax associated with a Mycoplasma sp and Arcanobacterium pyogenes was diagnosed at necropsy in a 1-month-old female Van kitten . The pleural cavity contained approximately 50 mL of blood-tinged, reddish-brown, nonodourous fluid bilaterally . Gram positive coccobacilli were seen in the exudate from necrotic plaques on the pleurae . Mycoplasma sp and A pyogenes were isolated from a sample of the fluid in the pleural cavity . The concomitant presence of Mycoplasma sp and A pyogenes could be considered another variation on the polymicrobial nature of pyothorax and associated pleural lesions in cats. J Am Dent Assoc, 2002 Jul, 133(7), 837 - 41 Evaluation of compressed air used in the dental operatory; Hubar JS et al.; BACKGROUND: The authors report the findings obtained when they quantitatively examined compressed air samples from air-water syringes located in different dental operatories at the Louisiana State University School of Dentistry fo r the presence of microbial contaminants . METHODS: Streams of air of 30 seconds' duration from air-water syringes were forced through sterile modified stainless steel membrane filter holders (Millipore, Millipore Corp., Bedford, Mass.), each containing a membrane filter (average pore diameter = 0.45 micrometers) . Each filter was aseptically removed, placed onto the surface of a Petri dish containing sheep blood agar and incubated under increased carbon dioxide tension at 37 C for 48 hours . The authors performed a count of the resultant microbial colonies, after which they microscopically examined the gram-stained organisms . RESULTS: Bacteria were detected in 24 percent of the samples . The number of colonies observed on the filters varied among the dental units . The air from only one of the dental units sampled repeatedly was found to be free of bacterial contaminants . This contrasted with other units for which one or more samples were found to be positive for microorganisms . The majority of colonies observed were pigmented . Microscopic examination of organisms from representative colonies revealed that most were either gram-positive cocci or gram-negative diplococci and tetrads . The results of the one-sample t test were found to be significant (t = 5.6, df = 98, P = .0001) . The 95 percent confidence interval was 0.15 to 0.32 . CONCLUSION: The results suggest that, at least statistically, a percentage of air lines will have bacteria present. Int J Syst Evol Microbiol, 2002 Jul, 52(Pt 4), 1263 - 6 Turicibacter sanguinis gen . nov., sp . nov., a novel anaerobic, Gram-positive bacterium; Bosshard PP et al.; An unknown, strictly anaerobic, Gram-positive, rod-shaped bacterium (strain MOL361T) was isolated from a blood culture of a febrile patient with acute appendicitis and characterized using phenotypic and molecular methods . Fatty acid analysis and biochemical examination indicated that the isolate most closely resembles members of the Gram-positive bacteria with low DNA G+C content . 16S rDNA sequencing revealed a relatively high overall similarity (97%) to an uncultured bacterium, but these two strains both exhibit low (<87%) 16S rDNA similarity to other bacteria . Phylogenetic analysis with different treeing methods showed that this strain forms a novel line of descent within the Gram-positive bacteria with low G+C content . Strain MOL361T is described as the type strain of a novel species within a new genus, Turicibacter sanguinis gen . nov., sp . nov. Int J Syst Evol Microbiol, 2002 Jul, 52(Pt 4), 1229 - 34 Microbacterium aerolatum sp . nov., isolated from the air in the 'Virgilkapelle' in Vienna; Zlamala C et al.; Three rod-shaped, Gram-positive strains were isolated from the air of the chapel 'Virgilkapelle' in Vienna . A representative of these three strains, strain V-73T, shared the highest 16S rDNA sequence similarities with members of the genus Microbacterium, in particular Microbacterium foliorum, Microbacterium testaceum, Microbacterium esteraromaticum, Microbacterium keratanolyticum and Microbacterium arabinogalactanolyticum . The strains displayed almost identical biochemical and physiological characteristics and showed no differences in their protein patterns obtained after SDS-PAGE . On the basis of Fourier-transform infra-red (FT-IR) spectra and genomic fingerprints, the three strains were grouped together and separated from the other relevant members of the genus Microbacterium . The chemotaxonomic characteristics analysed, including polar lipids, quinone systems, cell wall composition and fatty acid profiles, were in good agreement with the characteristics described for the genus Microbacterium . The G+C content of the DNAs was determined to be in the narrow range 69.3-69.7 mol % . The results of DNA-DNA hybridization, biochemical/physiological characterization, ERIC-PCR-generated genomic fingerprints and FT-IR spectra demonstrated that the three isolates represent a novel species of the genus Microbacterium . The name Microbacterium aerolatum sp . nov . is proposed for the novel species, of which strain V-73T (= DSM 14217T = CCM 4955T) is the type strain. Int J Syst Evol Microbiol, 2002 Jul, 52(Pt 4), 1211 - 6 Agrococcus baldri sp . nov., isolated from the air in the 'Virgilkapelle' in Vienna; Zlamala C et al.; Five coccoid, Gram-positive strains were isolated from the air of the 'Virgilkapelle' in Vienna . A representative of these five strains, V-108T, shared 99.0 and 98.4% 16S rDNA sequence similarity, respectively, with Agrococcus jenensis DSM 9580 and Agrococcus citreus DSM 12453T . Colonies of the five strains were white when grown in the dark and turned yellow in the light . The strains displayed highly similar biochemical and physiological characteristics and showed only small differences in their protein patterns obtained after SDS-PAGE . Based on Fourier-transform infra-red (FT-IR) spectra, the five strains were grouped together and separated from the other members of the genus, A . jenensis and A . citreus . Chemotaxonomic characteristics analysed from selected members of the five isolates, including polar lipids, quinone systems, polyamine patterns, cell wall composition and fatty acid profiles, were in good agreement with those of the two species of the genus Agrococcus described to date . The G+C content of the genomic DNA was determined to be within the narrow range of 73.8-74.9 mol% . The results of DNA-DNA hybridization with A . citreus DSM 12453T and A . jenensis DSM 9580T, as well as differences in biochemical/physiological characteristics, peptidoglycan composition, fatty acids, polar lipid profiles and FT-IR spectra, demonstrated that the five isolates represent a novel species of the genus Agrococcus . The name Agrococcus baldri sp . nov . is proposed for the novel species, of which strain V-108T (= DSM 14215T = CCM 4953T) is the type strain. Int J Syst Evol Microbiol, 2002 Jul, 52(Pt 4), 1145 - 50 Nesterenkonia lacusekhoensis sp . nov., isolated from hypersaline Ekho Lake, East Antarctica, and emended description of the genus Nesterenkonia; Collins MD et al.; An aerobic and heterotrophic isolate, designated IFAM EL-30T, was obtained from hypersaline Ekho Lake (Vestfold Hills, East Antarctica) . The isolate consisted of Gram-positive cocci or short rods which occasionally exhibited branching . The organism was moderately halotolerant, required thiamin.HCI and was stimulated by biotin and nicotinic acid . It grew well with glucose, acetate, pyruvate, succinate, malate or glutamate, and hydrolysed DNA but not gelatin, starch or Tween 80 . Nitrate was aerobically reduced to nitrite . Chemical analysis revealed diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and an unidentified glycolipid as the major polar lipids . The cellular fatty acids were predominantly of the anteiso and iso methyl-branched types, and the major menaquinone6 were MK-7 and MK-8 . The peptidoglycan type was A4alpha, L-Lys-L-Glu . The DNA base ratio was 66.1 mol% G+C . Comparisons of 16S rRNA gene sequences showed that the unidentified organism was phylogenetically closely related to Nesterenkonia halobia, although a sequence divergence value of > 3% demonstrated that the organism represents a different species . On the basis of phenotypic and genotypic evidence, it is proposed that the unknown bacterium be designated as a new species of the genus Nesterenkonia, namely Nesterenkonia lacusekhoensis sp . nov., the type strain being IFAM EL-30T (= DSM 12544T = CIP 107030T) . An emended description of the genus Nesterenkonia is given. Int J Syst Evol Microbiol, 2002 Jul, 52(Pt 4), 1133 - 9 Gordonia westfalica sp . nov., a novel rubber-degrading actinomycete; Linos A et al.; A cis-1,4-polyisoprene-degrading bacterium (strain Kb2T) was isolated from foul water taken from the inside of a deteriorated automobile tyre found on a farmer's field in Westfalia, Germany . The strain was aerobic, Gram-positive, exhibited orange smooth and rough colonies on complex nutrient agar, produced elementary branching hyphae that fragmented into rod/coccus-like elements and showed chemotaxonomic markers which were consistent with its classification within the genus Gordonia, i.e . the presence of mesodiaminopimelic acid, arabinose and galactose in whole-cell hydrolysates (cell-wall chemotype IV), N-glycolylmuramic acid in the peptidoglycan wall, a fatty-acid pattern composed of unbranched saturated and monounsaturated fatty acids plus tuberculostearic acid, mycolic acids comprising 56-60 carbon atoms and MK-9(H2) as the only menaquinone . The 16S rDNA sequence of strain Kb2T was found to be most similar to the 16S rDNA sequences of the type strains of Gordonia alkanivorans (DSM 44369T) and Gordonia nitida (KCTC 0605BPT) . However, DNA-DNA relatedness data showed that strain Kb2T ( =DSM 44215T NRRL B-24152T) could be distinguished from these two species and represented a new species within the genus Gordonia, for which the name Gordonia westfalica is proposed. Int J Syst Evol Microbiol, 2002 Jul, 52(Pt 4), 1081 - 7 Methanobrevibacter acididurans sp . nov., a novel methanogen from a sour anaerobic digester; Savant DV et al.; A novel acid-tolerant, hydrogenotrophic methanogen, isolate ATMT, was obtained from an enrichment performed at pH 5.0 using slurry from an acidogenic digester running on alcohol distillery waste . The original pH of the slurry was 5.7 and the volatile fatty acid concentration was 9000 p.p.m . Cells of isolate ATMT were Gram-positive, non-motile and 0.3-0.5 microm in size . They did not form spores . The isolate could grow in the pH range 5.0-7.5, with maximum growth at pH 6.0 . The optimum temperature for growth was 35 degrees C . Formate, acetate, methanol, trimethylamine, 2-propanol and 2-butanol were not utilized as growth substrates . Rumen fluid and acetate were required for growth on H2/CO2 . Coenzyme M and 2-methylbutyric acid were not required in the presence of rumen fluid . 16S rDNA sequence analysis confirmed the signature sequence of the genus Methanobrevibacter . Morphological and biochemical characteristics of the isolate, together with the 16S rDNA sequence analysis, clearly revealed that the isolate could not be accommodated within any of the existing species of the genus Methanobrevibacter . Therefore, it is proposed that a novel species of the genus Methanobrevibacter should be created for this isolate, Methanobrevibacter acididurans sp . nov., and the type strain is Am J Kidney Dis, 2002 Aug, 40(2), 373 - 80 Comparison of clinical outcome and ease of handling in two double-bag systems in continuous ambulatory peritoneal dialysis: a prospective, randomized, controlled, multicenter study; Li PK et al.; BACKGROUND: We performed a prospective, randomized, controlled, multicenter study on the use of two double-bag disconnect systems: Stay-Safe (SS; Fresenius Deutschland GmbH2) and Ultrabag (UB; Baxter Healthcare, Deerfield, IL) to assess the ease of handling, peritonitis rate, exit-site infection rate, and clinical outcome . METHODS: We enrolled 110 new continuous ambulatory peritoneal dialysis (CAPD) patients; 55 patients were randomized to SS treatment, and 55 patients, to UB treatment . RESULTS: Patients using the UB and SS systems were followed up for 946 and 846 patient-months, respectively . There were 21 episodes of peritonitis in 18 patients in the UB group and 23 episodes in 18 patients in the SS group . No significant difference was observed in peritonitis rates between the two systems, which were 45 and 36.8 patient-months per episode for the UB and SS groups, respectively . At 12 months, 82.1% of patients in the UB group and 72.1% in the SS group were free of peritonitis; at 18 months, 71.1% and 62.2% were free of peritonitis for the UB and SS groups, respectively (P = 0.559) . Gram-positive organisms accounted for 28.6% of infections in the UB group and 39.1% in the SS group . Exit-site infection rates were one episode per 21 patient-months versus 19.2 patient-months in the UB and SS groups, respectively (P = 0.743) . Patients perceived SS as easier to handle in 4 of the 13 steps immediately post-CAPD training . However, there was no significant difference in rankings between the two systems after 1 month of adaptation . Median training periods were 4 and 5 days for the SS and UB groups, respectively (P = 0.640) . CONCLUSION: The two double-bag systems (UB and SS) have similar incidences of peritonitis and exit-site infection . Both systems showed comparably good clinical outcome . The SS system is easier to learn during the initial training period, but the difference is not significant after 1 month's adaptation . Appl Environ Microbiol, 2002 Aug, 68(8), 4145 - 7 Noninvasive measurement of bacterial intracellular pH on a single-cell level with green fluorescent protein and fluorescence ratio imaging microscopy; Olsen KN et al.; We show that a pH-sensitive derivative of the green fluorescent protein, designated ratiometric GFP, can be used to measure intracellular pH (pHi) in both gram-positive and gram-negative bacterial cells . In cells expressing ratiometric GFP, the excitation ratio (fluorescence intensity at 410 and 430 nm) is correlated to the pHi, allowing fast and noninvasive determination of pHi that is ideally suited for direct analysis of individual bacterial cells present in complex environments. Appl Environ Microbiol, 2002 Aug, 68(8), 3969 - 77 A single Ala139-to-Glu substitution in the Renibacterium salmoninarum virulence-associated protein p57 results in antigenic variation and is associated with enhanced p57 binding to chinook salmon leukocytes; Wiens GD et al.; The gram-positive bacterium Renibacterium salmoninarum produces relatively large amounts of a 57-kDa protein (p57) implicated in the pathogenesis of salmonid bacterial kidney disease . Antigenic variation in p57 was identified by using monoclonal antibody 4C11, which exhibited severely decreased binding to R . salmoninarum strain 684 p57 and bound robustly to the p57 proteins of seven other R . salmoninarum strains . This difference in binding was not due to alterations in p57 synthesis, secretion, or bacterial cell association . The molecular basis of the 4C11 epitope loss was determined by amplifying and sequencing the two identical genes encoding p57, msa1 and msa2 . The 5' and coding sequences of the 684 msa1 and msa2 genes were identical to those of the ATCC 33209 msa1 and msa2 genes except for a single C-to-A nucleotide mutation . This mutation was identified in both the msa1 and msa2 genes of strain 684 and resulted in an Ala(139)-to-Glu substitution in the amino-terminal region of p57 . We examined whether this mutation in p57 altered salmonid leukocyte and rabbit erythrocyte binding activities . R . salmoninarum strain 684 extracellular protein exhibited a twofold increase in agglutinating activity for chinook salmon leukocytes and rabbit erythrocytes compared to the activity of the ATCC 33209 extracellular protein . A specific and quantitative p57 binding assay confirmed the increased binding activity of 684 p57 . Monoclonal antibody 4C11 blocked the agglutinating activity of the ATCC 33209 extracellular protein but not the agglutinating activity of the 684 extracellular protein . These results indicate that the Ala139-to-Glu substitution altered immune recognition and was associated with enhanced biological activity of R . salmoninarum 684 p57. OMICS, 2002, 6(2), 199 - 206 Genome comparison of Mycobacterium tuberculosis and other bacteria; Fu LM et al.; The availability of the complete genome sequence of Mycobacterium tuberculosis allows its phylogenetic analysis based on the whole genome rather than single genes . As a genome-based tree is more representative of whole organisms and less inconsistent than single-gene trees, it could provide a better index for interpretation and inference about the origin and nature of species . The standard bacterial phylogeny based on 16S ribosomal RNA sequence comparison shows that M . tuberculosis is more related to Gram-positive than to Gram-negative bacteria . Our results based on genome comparison in terms of shared orthologous genes challenge this implication . We demonstrate that M . tuberculosis is more related to Gram-negative than to Gram-positive bacteria by a quantitative analysis on the genome tree . The numerical distance data derived from genome comparison and those from 16S rRNA comparison show high significant correlation, implying that conserved gene content carries a strong phylogenetic signature in evolution. Planta Med, 2002 Jul, 68(7), 631 - 4 Quinoline alkaloids, coumarins and volatile constituents of Helietta longifoliata; de Moura NF et al.; A new quinoline alkaloid named helietidine ( 1), and seven known compounds ( 2 - 8) have been isolated from the stem barks of Helietta longifoliata . The structures of the new and the known compounds were established on the basis of spectral evidence, especially by 2D NMR ( 1H- 1H COSY, NOESY, HMQC, HMBC) . In addition, the volatile constituents of H . longifoliata leaves were analysed by gas chromatography-mass spectrometry (GC-MS) . Twenty-five constituents were identified representing ca . of 96 % of the oil, and limonene (17.50 %), germacrene D (16.60 %), elemol (11.81 %), bicyclogermacrene (11.67 %), guaiol (11.53), and epi-alpha-bisabolol (7.24 %) were the most abundant components . The oil was bioactive against some Gram-positive and Gram-negative bacteria, as revealed by bioautography. J Vet Intern Med, 2002 Jul-Aug, 16(4), 464 - 71 Actinobacillus sp . bacteremia in foals: clinical signs and prognosis; Stewart AJ et al.; Medical records of 101 blood culture-confirmed bacteremic foals were reviewed to determine whether foals with Actinobacillus sp . bacteremia are affected at an earlier age, have more severe signs of disease, and have a worse prognosis than do foals with bacteremia of other causes . Thirty percent (30/101) of bacteremic foals had Actinobacillus sp . cultured, and these were 2 times more likely to die (crude odds ratio {OR(CR)} 0.8, 4; P = .14), with a survival rate of 43% (13/30) compared to the overall survival rate of 55% (56/101) . When compared to other bacteremic foals, foals with actinobacillosis were 7 times more likely to have been sick from birth (adjusted odds ratio {OR(ADJ)} 2, 26; P = .003) and 6 times more likely to have diarrhea (OR(ADJ) 1, 22; P = .009) . By bivariate analysis . foals with Actinobacillus sp . bacteremia were 5 times more likely to have a sepsis score >11 (OR(CR) 1, 18; P = .007), 6 times more likely to be obtunded (OR(CR) 2, 20; P = .005), and 3 times more likely to have pneumonia (OR(CR) 1, 7; P = .03) . Furthermore, Actinobacillus sp . bacteremic foals were 27 times more likely to have a segmented neutrophil count <3.3 X 10(9) cells/L (OR(ADJ) 4, 166: P < .0001) and were 4.5 times more likely to have a band neutrophil count >0.46 x 10(9) cells/L (OR(ADJ) 1, 17; P = .02) when compared to foals that had bacteremia caused by either gram-negative enteric or gram-positive organisms . Sepsis score was < or = 11 in 49% (29/59) of bacteremia foals aged <13 days for which a discernible sepsis score was calculable . Results of this study should improve the diagnostic sensitivity of clinical examinations of neonatal foals, thereby facilitating treatment decisions. J Biol Chem, 2002 Oct 4, 277(40), 37339 - 48 Epub 2002 Jul 22. Structural organization of the protein-tyrosine autokinase Wzc within Escherichia coli cells; Doublet P et al.; Protein Wzc from Escherichia coli is a member of a newly defined family of protein-tyrosine autokinases that are essential for surface polysaccharide production in both Gram-negative and Gram-positive bacteria . Although the catalytic mechanism of the autophosphorylation of Wzc was recently described, the in vivo structural organization of this protein remained unclear . Here, we have determined the membrane topology of Wzc by performing translational fusions of lacZ and phoA reporter genes to the wzc gene . It has been shown that Wzc consists of two main structural domains: an N-terminal domain, bordered by two transmembrane helices, which is located in the periplasm of cells, and a C-terminal domain, harboring all phosphorylation sites of the protein, which is located in the cytoplasm . In addition, it has been demonstrated for the first time that Wzc can oligomerize in vivo to form essentially trimers and hexamers . Cross-linking experiments performed on strains expressing various domains of Wzc have shown that the cytoplasmic C-terminal domain is sufficient to generate oligomerization of Wzc . Mutant proteins, modified in either the ATP-binding site or the different phosphorylation sites, i.e . rendered unable to undergo autophosphorylation, have appeared to oligomerize into high molecular mass species identical to those formed by the wild-type protein . It was concluded that phosphorylation of Wzc is not essential to its oligomerization . These data, connected with the phosphorylation mechanism of Wzc, may be of biological significance in the regulatory role played by this kinase in polysaccharide synthesis. Z Naturforsch {C}, 2002 May-Jun, 57(5-6), 434 - 9 Chemical composition and biological activity of the essential oils of Senecio aegyptius var . discoideus Boiss; El-Shazly A et al.; The essential oil of Senecio aegyptius var . discoideus flowers, leaves, stems and roots were isolated by hydrodistillation . Analysis of the oils by capillary GLC and GLC-mass spectrometry were performed and 34 out of 37 compounds were identified . The main component was isolated and characterized as 1,10-epoxyfuranoeremophilane using a combination of GLC, GLC-MS, and NMR analyses . The oils of flowers, leaves and stems were rich in monoterpene hydrocarbons while the root oil mainly contains furanoeremophilanes . Flower and leaf volatile oils showed significant level of antifungal activity against C . albicans, moderate effect against Gram positive bacteria, however, it has weak activity against Gram negative bacteria . The isolated sesquiterpene (1,10-epoxyfuranoeremophilane) exhibited substantial inhibitory activity against Gram negative bacteria. Clin Nutr, 2002 Jun, 21(3), 207 - 11 Prevention and treatment of implanted central venous catheter (CVC) - related sepsis: a report after six years of home parenteral nutrition (HPN); Santarpia L et al.; Catheter-related sepsis is a serious and common complication in patients receiving home parenteral nutrition (HPN) . Prevention measures, prevalence of infections, types of agents and implanted central venous catheters (CVC), effectiveness of antibiotic therapy have been evaluated in 221 patients consecutively followed in our unit from January 1995 to December 2000 . The clinical diagnosis of catheter-related infection was made using well-defined criteria . Patients were divided into two groups: A and B, receiving instructions with different modalities: standard (A) and detailed (B), respectively . Sixty CVC-related sepsis occurred in 32 (14%) patients . A multivariate analysis showed that the duration of HPN (P<0.001; OR=0.9), type of catheter (P=0.009; OR=0.12) and type of disease (P=0.033; OR=4.92) significantly influence catheter infection . The type of implanted CVC (159 port-a-cath in 153 patients and 71 tunnelled in 68) seems to affect the infection rate, this being lower in tunnelled (P=0.03) . Infection rate was lower in B vs A group (P<0.001) with all types of catheters, suggesting the preventive role of very careful training . In particular, the incidence of CVC-related sepsis was 6/1000 days of HPN (i.e . 6/1000 days of catheterization) in Group A and 3/1000 in Group B . Systemic and antibiotic lock therapy was performed with an 83% successful rate . Gram-positive bacteria were the most frequent CVC infection agents, which are usually eradicated by antibiotic therapy lasting 7 days. Intensive Care Med, 2002 Jul, 28(7), 824 - 33 Epub 2002 May 30. Clinical implications of antibiotic-induced endotoxin release in septic shock; Lepper PM et al.; Antibiotic-induced release of bacterial cell wall components can have immediate adverse effects for the patient . This article reviews the data on endotoxin release after initiation of antibiotic therapy and its role in the pathogenesis of sepsis and septic shock . Antibiotics differ in their potential to liberate endotoxins from bacterial cell walls . When used for treatment of systemic Gram-negative infection, some classes of beta-lactam antibiotics lead to markedly increased levels of free endotoxins while treatment with carbapenems and aminoglycosides produces relatively low amounts of endotoxins . Antibiotics that induce the formation of long, aberrant bacterial cells before effectively killing the microorganisms show the highest degree of endotoxin liberation . There is increasing evidence from animal models and clinical studies of sepsis that the antibiotic-mediated release of biologically active cell wall components derived from Gram-positive, Gram-negative or fungal organisms is associated with a rapid clinical deterioration. J Comp Physiol {B}, 2002 Jul, 172(5), 441 - 6 Epub 2002 Jun 14. Effects of nitric oxide synthase inhibitors on the febrile response to muramyl dipeptide and lipopolysaccharide in rats; Kamerman PR et al.; We have administered aminoguanidine, a relatively specific inhibitor of inducible nitric oxide synthase, and N-nitro-L-arginine methyl ester (L-NAME), an unspecific nitric oxide synthase inhibitor, to rats made febrile with the gram-positive pyrogen, muramyl dipeptide and gram-negative pyrogen, lipopolysaccharide . Sprague-Dawley rats, housed individually at approximately 25 degrees C with a 12:12 h light:dark cycle (lights on 0700 hours), were injected (at 0900 hours) intraperitoneally with 50 mg/kg aminoguanidine, 25 mg/kg or 50 mg/kg L-NAME, and intramuscularly with 500 microg/kg muramyl dipeptide or 100 microg/kg lipopolysaccharide . Pyrogen injections were spaced at least 14 days apart . Body temperature was measured throughout the study in unrestrained animals using radio-telemetry . Neither muramyl dipeptide nor lipopolysaccharide-induced fevers were affected by aminoguanidine . However, L-NAME administration inhibited muramyl dipeptide and lipopolysaccharide-induced fevers, but only for the 1st 2-4 h of the fevers (two-way ANOVA, P<0.05) . After the initial inhibition, lipopolysaccharide fevers developed normally . Therefore, constitutively expressed nitric oxide synthase appears to be involved in the initial phases of fever genesis of gram-negative and gram-positive fevers in rats . On the other hand, inducible nitric oxide synthase appears not to play a role in these fevers. Infection, 2002 Jun, 30(3), 178 - 80 Actinomyces neuii and neonatal sepsis; Mann C et al.; Actinomyces neuii belongs to the coryneform bacteria . In the case presented here this gram-positive rod had to be considered the pathogen causing not only the chorioam nionitis but also the neonatal sepsis . Conventional therapeutic regimes are effective due to the high sensitivity of A . neuii to beta-lactam antibiotics. Am J Cardiol, 2002 Jul 15, 90(2), 119 - 23 Significance of peptidoglycan, a proinflammatory bacterial antigen in atherosclerotic arteries and its association with vulnerable plaques; Laman JD et al.; Peptidoglycan (PG) is a major component of the cell wall of gram-positive bacteria that is abundantly present in all human mucosa . PG is a functional lipopolysaccharide analog that binds to CD14 on macrophages and induces proinflammatory cytokine production and metalloproteinases . We investigated the hypothesis that bacterial PG is present in atherosclerotic tissue . In addition, plaque phenotypes were characterized in relation to presence of PG . Immunohistology of carotid (n = 15) and femoral (n = 6) endarterectomy specimens revealed the presence of PG in the cytoplasm of cells located in plaques . PG was detected in 14 of 15 carotid arteries and 5 of 6 femoral arteries . From the 14 coronary arteries, 31 atherosclerotic segments were selected . PG was detected within 19 of 31 of these coronary segments . Western blot demonstrated the presence of the toll-like receptor (TLR-2), the co-receptor for PG, in coronary artery tissue . The number of PG-containing cells in coronary arteries was significantly higher when the histologic features of plaque vulnerability were evident . Inflammation of the cap or shoulder was observed in 11 of 19 PG-positive versus 2 of 12 PG-negative segments (p = 0.023) . More than 50% of the plaque area consisted of atheroma in 7 of 19 PG-positive segments and 0 of 12 PG-negative segments (p = 0.025) . Heavy smooth muscle cell staining occurred in the plaque cap and shoulder in 3 of 19 PG-positive segments versus 9 of 12 PG-negative segments . Proinflammatory bacterial PG and its co-receptor have been observed in atherosclerotic arteries, in association with the vulnerable plaque phenotype. Science, 2002 Jul 5, 297(5578), 114 - 6 Activation of Drosophila Toll during fungal infection by a blood serine protease; Ligoxygakis P et al.; Drosophila host defense to fungal and Gram-positive bacterial infection is mediated by the Spaetzle/Toll/cactus gene cassette . It has been proposed that Toll does not function as a pattern recognition receptor per se but is activated through a cleaved form of the cytokine Spaetzle . The upstream events linking infection to the cleavage of Spaetzle have long remained elusive . Here we report the identification of a central component of the fungal activation of Toll . We show that ethylmethane sulfonate-induced mutations in the persephone gene, which encodes a previously unknown serine protease, block induction of the Toll pathway by fungi and resistance to this type of infection. J Immunol, 2002 Jul 15, 169(2), 913 - 9 Production of granulocyte colony-stimulating factor in the nonspecific acute phase response enhances host resistance to bacterial infection; Noursadeghi M et al.; Mice mounting an acute phase response, induced by sterile inflammation after a single s.c . injection of casein 24 h beforehand, were remarkably protected against lethal infection with Gram-positive or Gram-negative bacteria . This was associated with enhanced early clearance of bacteremia, greater phagocytosis and oxidative burst responses by neutrophils, and enhanced recruitment of neutrophils into tissues compared with control, nonacute phase mice . Casein-induced inflammation was also associated with increased concentrations of G-CSF in serum, and administration of neutralizing Ab to this cytokine completely abrogated protection against Escherichia coli infection after casein pretreatment . Injection of recombinant murine G-CSF between 3 and 24 h before infection conferred the same protection as casein injection . In contrast, the casein-induced acute phase response affected neither serum values of TNF-alpha, IL-1 beta, or IL-6 after E . coli infection nor susceptibility to LPS toxicity . Furthermore, protection against infection was unaffected in IL-1R knockout mice, which have deficient acute phase plasma protein responses, or after nonspecific inhibition of acute phase protein synthesis by D-galactosamine or specific depletion of complement C3 by cobra venom factor . Increased production of G-CSF in the acute phase response is thus a key physiological component of host defense, and pretreatment with G-CSF to prevent bacterial infection in at-risk patients now merits further study, especially in view of increasing bacterial resistance to antibiotics. J Environ Monit, 2002 Jun, 4(3), 377 - 82 Evaluation of PCR primers and PCR conditions for specific detection of common airborne fungi; Wu Z et al.; We examined the selectivity of 53 sets of primers for environmental monitoring of indoor air quality . Thirty-six fungal strains, representing 26 species from 14 genera of commonly occurring fungi, and 16 different bacterial strains, representing both gram-negative and gram-positive species, were included in the experiment . We verified the specificity of 28 of the 53 sets of primers, which were classified as universal fungal, universal bacterial, group or species specific . The PCR conditions required for optimal specificity were also determined . These results can serve as a guide for the step-wise PCR-based detection and identification of airborne fungi commonly found in indoor environments. Appl Environ Microbiol, 2002 Jul, 68(7), 3238 - 42 Airborne microbial flora in a cattle feedlot; Wilson SC et al.; A total of 1,408 cattle held in eight commercial feedlot pens were used to examine the quantity and diversity of microorganisms in cattle feedlot air . The effect of two feeding patterns on the generation of airborne dust and the total numbers of microorganisms was also examined (four feedlot pens/treatment) . Microbial samples were collected, and dust particles that were 2.5 microm or less in diameter were measured with a Dustrak monitor during the evening dust peak for 4 days at sites both upwind and downwind of the feedlot pens . An Andersen biological cascade sampler was employed with different medium and incubation combinations for the capture and identification of bacteria and fungi . The results showed that when bacteria were considered, only nonpathogenic gram-positive organisms were recovered . However, gram-negative bacteria may have been present in a viable but nonculturable state . Fungi were recovered in smaller numbers than bacteria, and none of the fungi were pathogenic . The Dustrak results showed that one feeding pattern resulted in cattle behavior that generated levels of downwind dust lower (P = 0.04) than the levels generated by the behavior resulting from the other feeding pattern . However, the Andersen sampler results showed that there were no differences between feeding patterns with regard to the total number or diversity of microorganisms . The disparity may have been due to the different operating principles of the two systems . The overall numbers of microorganisms recovered were lower than those reported in studies of intensively housed farm animals in which similar recovery techniques were used. J Biol Chem, 2002 Sep 13, 277(37), 34264 - 70 Epub 2002 Jun 19. FEEL-1, a novel scavenger receptor with in vitro bacteria-binding and angiogenesis-modulating activities; Adachi H et al.; Employing the expression cloning technique, we cloned a novel scavenger receptor that is structurally unrelated to other scavenger receptors . The cloned receptor contained fasciclin (Fas-1), epidermal growth factor (EGF)-like, laminin-type EGF-like, and link domains . Based on the domain structures, we temporarily named it FEEL-1 (fasciclin, EGF-like, laminin-type EGF-like, and link domain-containing scavenger receptor-1) . A data base search suggested the presence of a paralogous gene of FEEL-1, the full-length cDNA of this gene was also cloned, and its nucleotide sequence was determined . The deduced amino acid sequence of the clone indicated that its domain organization is similar to FEEL-1, and we named this clone FEEL-2 . The effect of monoclonal antibodies against FEEL-1 indicated that FEEL-1 is the major receptor for 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbo-cyanine perchlorate (DiI)-labeled acetylated low density lipoprotein (DiI-Ac-LDL) in human umbilical vein endothelial cells . Reverse transcription and PCR analysis revealed that both FEEL-1 and FEEL-2 were expressed in several tissues and expressed highly in the spleen and lymph node . On the other hand, only FEEL-1 was expressed in mononuclear cells, particularly resting CD14(+) cells . The transient expression of FEEL-1 and FEEL-2 in Chinese hamster ovary cells demonstrated that both FEELs could bind to DiI-Ac-LDL . Both receptors were also found to bind to Gram-negative and Gram-positive bacteria . These results suggest that FEELs play important roles in the defense mechanisms against bacterial infection . Finally, the phenotypic effect of the inhibition of FEEL-1 on vascular remodeling was tested in vitro using the Matrigel tube formation assay, and we found a marked reduction in the degree of cell-cell interaction in anti-FEEL-1 monoclonal antibody-treated cells, suggesting the role of this receptor in angiogenesis. FEMS Microbiol Lett, 2002 Jun 4, 211(2), 169 - 73 Alpha-proteobacteria cultivated from marine sponges display branching rod morphology; Olson JB et al.; Most isolates recovered from marine environments are Gram-negative proteobacteria, even with the use of various media and media additions to enhance recoverability . Cultivation studies with two genera of deep-water sponges yielded nine isolates that demonstrated bulbous branching rod morphology, which is usually associated with microorganisms staining Gram-positive . Gram reactions indicated that the isolates were Gram-negative, which was confirmed by partial 16S rDNA sequencing . All nine isolates were shown to be alpha-proteobacteria most closely related to other alpha-proteobacteria isolated from various sponges. Int J Clin Pract, 2002 May, 56(4), 237 - 40 Surveillance study of bacteraemic episodes in febrile neutropenic children; Ariffin H et al.; We prospectively studied the type, frequency and outcome of infections in 513 patients with 762 consecutive episodes of febrile neutropenia (FN) over a five-year period between 1995 and 1999 in a single paediatric oncology unit . The findings were then compared with a similar study carried out in our unit between 1990 and 1994 . The types of bacterial isolates and sensitivity patterns were also studied to identify trends and to gauge the suitability of antibiotics chosen for empirical therapy . Bacteraemia was documented in 35.4% of FN episodes, although 70% of patients did not have an obvious site of sepsis . The majority of isolates (61.9%) were gram-negative bacteria, a consistent finding throughout the study period . Resistance to ceftazidime, amikacin and imipenem among gram-negative bacteria was 26.3%, 21.2% and 0.7%, respectively . Methicillin resistance among gram-positive bacteria was 26.3%, while no vancomycin-resistant bacteria were encountered . There were 36 sepsis-related deaths . Factors associated with a fatal outome were prolonged capillary refill time, hypotension, fever above 39 degrees C and pneumonia . Rapid neutrophil recovery was associated with a good prognosis . A change to our current choice of empirical antibiotics for FN, comprising ceftazidime/ceftriaxone and amikacin appears necessary because of the relatively high resistance rates found. Appl Microbiol Biotechnol, 2002 Jun, 59(1), 86 - 90 Epub 2002 Apr 12. Simultaneous detection and removal of organomercurial compounds by using the genetic expression system of an organomercury lyase from the transposon Tn MERI1; Narita M et al.; Using a newly identified organomercury lyase gene (merB3) expression system from Tn MERI1