Gram positive

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, the mercury resistance transposon first found in Gram-positive bacteria, a dual-purpose system to detect and remove organomercurial contamination was developed . A plasmid was constructed by fusing the promoterless luxAB genes as bioluminescence reporter genes downstream of the merB3 gene and its operator/promoter region . Another plasmid, encoding mer operon genes from merR1 to merA, was also constructed to generate an expression regulatory protein, MerR1, and a mercury reductase enzyme, MerA . These two plasmids were transformed into Escherichia coli cells to produce a biological system that can detect and remove environmental organomercury contamination . Organomercurial compounds, such as neurotoxic methylmercury at nanomolar levels, were detected using the biomonitoring system within a few minutes and were removed during the next few hours.

Eur J Clin Microbiol Infect Dis, 2002 May, 21(5), 373 - 8 Epub 2002 May 17.
Differential fluorescent staining method for detection of bacteria in blood cultures, cerebrospinal fluid and other clinical specimens; Fazii P et al.; The aim of this study was to evaluate a differential staining method to distinguish gram-positive from gram-negative bacteria in fluorescence . The method is based on two fluorochromes, one acting in the wavelength of red, i.e . the acridine orange, and another acting in the wavelength of green, i.e . the fluorescein . With this method, gram-positive bacteria appear yellow and gram-negative bacteria appear green . In view of the importance of a rapid aetiological diagnosis in cases of septicaemia, the differential staining method in fluorescence was compared with Gram stain for the detection of bacteria in blood . Of 5,820 blood cultures entered into the study and identified by the Bactec 9120 fluorescent series instrument (Becton Dickinson Europe, France), 774 were positive . Of the 774 positive cultures, 689 yielded only a single organism . The differential staining method in fluorescence detected 626 of the 689 cultures, while Gram stain detected 468 . On the basis of these results, the sensitivity of the differential staining method in fluorescence was 90.9%, while that of Gram stain was 67.9% . The difference between the two methods was statistically significant ( P<0.001) . The differential fluorescent staining method was more sensitive than Gram stain in the detection of bacteria in blood cultures during the incubation period . This technique provides a rapid, simple and highly sensitive staining method that can be used in conjunction with subculture methods . Whereas subculture requires an incubation period of 18-24 h, the fluorescent staining technique can detect bacteria on the same day that smears are prepared and examined . The differential fluorescent staining method was also evaluated for its ability to detect microorganisms in cerebrospinal fluid and other clinical specimens . The microorganisms were easily detected, even when bacterial counts in the specimens were low.

Tunis Med, 2002 Jan, 80(1), 37 - 9
{Actinomycosis: report of 21 cases . Experience of the Anatomy and Cytopathologic Pathology Laboratory of the Rabta Hospital}; Bellil K et al.; Actinomycosis is a chronic suppurative granulomatous disease caused by an anaerobic Gram positive germ, Actinomyces . From 1994 to 1999, 21 cases of actinomycosis have been diagnosed in our laboratory . The main localisations were cervicofacial and gynecologic the diagnosis relies on histopathologic examination that shows actinomycosic granules.

J Appl Microbiol, 2002, 93(1), 122 - 33
Transformation and expression of an anaerobic fungal xylanase in several strains of the rumen bacterium Butyrivibrio fibrisolvens; Gobius KS et al.; AIMS: To obtain reliable transformation of a range of Butyrivibrio fibrisolvens strains and to express a Neocallimastix patriciarum xylanase gene in the recipients . METHODS AND RESULTS: Eight strains (H17c, E14, LP1309, LP1028, AR11a, OB156, LP210B and LP461A) of Bu . fibrisolvens were transformed by the Gram-positive vector pUB110 . A xylanase expression/secretion cassette containing Bu . fibrisolvens promoter and signal peptide elements fused to catalytic domain II of the N . patriciarum xylanase A cDNA (xynANp) was inserted into pUB110 to create the plasmid pUBxynA . pUBxynA was used to transform seven of the Bu . fibrisolvens strains transformed by pUB110 . In strain H17c pUBxynA, which produced native xylanase, 2.46 U mg-1 total xylanase activity was produced with 45% extracellular xylanase . In strain H17c pUMSX, 0.74 U mg-1 total xylanase activity was produced with 98% extracellular xylanase . H17c pUBxynA exhibited increased (28.7%) degradation of neutral detergent fibre compared with unmodified H17c; however, progressive loss of pUBxynA was observed in long-term cultivation . CONCLUSIONS: A stable transformation system was developed that was applicable for a range of Bu . fibrisolvens strains and high levels of expression of a recombinant xylanase were obtained in H17c which lead to increased fibre digestion . SIGNIFICANCE AND IMPACT OF THE STUDY: This stable transformation system with the accompanying recombinant plasmids will be a useful tool for further investigation aimed at improving ruminal fibre digestion.

Acta Ophthalmol Scand, 2002 Jun, 80(3), 258 - 61
Endophthalmitis following cataract surgery in Sweden.The 1998 national prospective survey; Montan P et al.; PURPOSE: To investigate the morbidity of postoperative endophthalmitis (POE) following cataract surgery in Sweden in 1998 . METHODS: Clinically presumed cases of POE were reported in a prospective survey in which all Swedish ophthalmic surgical units except one had agreed to participate . Data on intraocular cultures and visual outcomes at 3 months after infection were supplied . Surgical cases that became infected were identified in the Swedish National Cataract Register, thereby enabling screening for various putative risk factors . RESULTS: The nationwide incidence of POE amounted to 58 cases out of 54 666 cataract operations, or 0.1% of surgical cases . The predominant aetiology was gram-positive bacteria, which comprised 57% of the material . Acrylic intraocular lenses were found to decrease the risk of POE significantly in comparison to hydrogel and polymethylmethacrylate lenses . CONCLUSIONS: The incidence of POE after cataract surgery in Sweden is similar to that currently reported elsewhere in the developed world . We hope that continued registration of cases of POE in Sweden will shed light on the possible influences of various prophylactic measures and different intraocular lens materials on the development of postoperative infection.

Microbiology, 2002 Jun, 148(Pt 6), 1871 - 80
Adhesion to cellulose of the Gram-positive bacterium Ruminococcus albus involves type IV pili; Rakotoarivonina H et al.; This study was aimed at characterizing a cell-surface 25 kDa glycoprotein (GP25) that was previously shown to be underproduced by a spontaneous adhesion-defective mutant D5 of Ruminococcus albus 20 . An antiserum against wild-type strain 20 was adsorbed with the mutant D5 to enrich it in antibodies 'specific' to adhesion structures of R . albus 20 . The resulting antiserum, called anti-Adh serum, blocked adhesion of R . albus 20 and reacted mainly with GP25 in bacterial and extracellular protein fractions of R . albus 20 . The N-terminal sequence of purified GP25 was identical to that of CbpC, a 21 kDa cellulose-binding protein (CBP) of R . albus 8 . The nucleotide sequence of the gp25 gene was determined by PCR and genomic walking procedures . The gp25 gene encoded a protein of 165 aa with a calculated molecular mass of 16940 Da that showed 72.9% identity with CbpC and presented homologies with type IV pilins of Gram-negative pathogenic bacteria . Negative-staining electron microscopy revealed fine and flexible pili surrounding R . albus 20 cells while mutant cells were not piliated . In addition, immunoelectron microscopy showed that the anti-Adh serum probing mainly GP25, completely decorated the pili surrounding R . albus 20, thereby showing that GP25 was a major pilus subunit . This study shows for the first time the presence of pili at the surface of R . albus and identifies GP25 as their major protein subunit . Though GP25 was not identified as a CBP, isolated pili were shown to bind cellulose . In conclusion, these pili, which belong to the family of type IV pili, mediate adhesion of R . albus 20 to cellulose.

CLAO J, 2002 Apr, 28(2), 66 - 8
Nocardia asteroides sclerokeratitis in a contact lens wearer; Sridhar MS et al.; PURPOSE: To report a case of sclerokeratitis caused by Nocardia asteroides in a soft contact lens wearer . CASE REPORT: A 65-year-old male presented with a 2-month history of a corneal ulcer in the left eye . He wore two weekly disposable soft contact lenses on an extended basis . He revealed his history of gardening before the onset of symptoms . On examination, his best-corrected visual acuity was 20/30 in the right eye and 20/400 in the left eye . In the left eye, there was conjunctival injection . His cornea showed multiple patchy infiltrates, with a feathery border that was raised and involved up to the midstroma . There was a 3+ anterior chamber reaction . Corneal scrapings were performed for smears and cultures . Topical 2% amikacin sulfate every half hour along with oral clarithromycin therapy was initiated . On follow-up, the sclera lesions worsened . RESULTS: Smears of corneal scrapings revealed gram-positive filamentous bacteria in Gram's stain . The cultures grew Nocardia asteroides . The patient was switched to trimethoprim-sulfamethoxazole (Bactrim DS, Roche Laboratories, Nutley, NJ) as the sclera was involved . The patient responded to treatment, and the infection resolved . When last seen, approximately 4 months after his initial presentation to us, his visual acuity was 20/40 in the affected eye . There was corneal scarring, and the adjacent sclera showed thinning . CONCLUSIONS: Nocardia sclerokeratitis can be associated with contact lens wear . Nocardia should be considered in the differential diagnosis of a corneal ulcer with an indolent progressive course with feathery infiltrates . Topical amikacin and systemic trimethoprim-sulfamethoxazole are effective drugs in the treatment of nocardial corneal infection with scleral involvement.

Int J Syst Evol Microbiol, 2002 May, 52(Pt 3), 933 - 8
Kineococcus radiotolerans sp . nov., a radiation-resistant, gram-positive bacterium; Phillips RW et al.; A gram-type positive, motile, coccus-shaped organism was isolated from a radioactive work area . Strain SRS30216T is an orange-pigmented bacterium that is catalase-positive, oxidase-negative and urease-negative . The orange pigment is most likely a carotenoid with absorption peaks at approximately 444, 471 and 501 nm . Cells normally grew in clusters, but individual, motile, flagellated cells were also observed . Growth of strain SRS30216T occurred at temperatures between 11 and 41 degrees C, between pH 5 and 9 and at NaCl concentrations up to and including 5% . Fatty acid composition was limited, with >90% of the fatty acids being anteiso 15:0 . Alkenes of 19-24 carbons in length were detected during examination of the neutral lipids . Strain SRS30216T demonstrated high levels of resistance to gamma-radiation and desiccation . The most closely related recognized species is Kineococcus aurantiacus RA 333T, which is 93% similar in 16S rDNA sequence . DNA-DNA hybridization revealed only 31% similarity between these two organisms . It is proposed that SRS30216T (= ATCC BAA-149T = DSM 14245T) represents the type strain of a novel species in the genus Kineococcus, Kineococcus radiotolerans sp . nov..

Int Microbiol, 2001 Dec, 4(4), 187 - 202
The branching order and phylogenetic placement of species from completed bacterial genomes, based on conserved indels found in various proteins; Gupta RS; The presence of shared conserved inserts and deletions (indels or signature sequences) in proteins provides a powerful means for understanding the evolutionary relationships among the Bacteria . Using such indels, all of the main groups within the Bacteria can be defined in clear molecular terms and it has become possible to deduce that they branched from a common ancestor in the following order: Low G + C gram-positive --> High G+C gram-positive --> Deinococcus Thermus --> Cyanobacteria --> Spirochetes --> Aquifex-Chlamydia-Cytophaga --> Proteobacteria-1 (epsilon, delta) --> Proteobacteria-2 (alpha) --> Proteobacteria-3 (beta) --> Proteobacteria -4 (gamma) . The usefulness of this approach for understanding bacterial phylogeny was examined here using sequence data from various completed bacterial genomes . By using 12 indels in highly conserved and widely represented proteins, the species from all 41 completed bacterial genomes were assigned to different groups; and the observed distribution of these indels in different species was then compared with that predicted by the signature sequence model . The presence or absence of these indels in various proteins in different bacteria followed the pattern exactly as predicted: and, in more than 450 observations, no exceptions or contradictions in the placement of indels were observed . These results provide strong evidence that lateral gene transfer events have not affected the genes containing these indels to any significant extent . The phylogenetic placement of bacteria into different groups based on signature sequences also showed an excellent correlation with the 16 S rRNA with 39 of the 41 species assigned to the same group by both methods . These results strongly vindicate the usefulness of the signature sequence approach to understanding phylogeny within the Bacteria and show that it provides a reliable and internally consistent means for the placement of bacterial species into different groups and for determining the relative branching order of the groups.

J Refract Surg, 2002 May-Jun, 18(3), 280 - 6
Microbial keratitis after laser in situ keratomileusis; Pushker N et al.; PURPOSE AND METHOD: To review the literature on microbial keratitis reported after laser in situ keratomileusis (LASIK) . RESULTS: Forty-one eyes have been reported to have microbial keratitis after LASIK . The causative organisms vary from gram positive bacteria to atypical mycobacteria, fungal, and viral pathogens . The infection is usually acquired intraoperatively, but may also be caused by postoperative contamination . A majority of the patients present within 72 hours of the surgery with an acute onset of symptoms . Management of microbial keratitis after LASIK includes aggressive topical fortified antibiotic therapy, irrigation of stromal bed with antibiotic solution after lifting the flap, and sending the scraping of the infiltrate for microbiological evaluation . The keratitis heals with scarring and a best spectacle-corrected visual acuity of 20/40 or better can be obtained in the majority of the patients . CONCLUSION: Microbial keratitis is a sight-threatening complication of LASIK.

Biometals, 2002 Jun, 15(2), 121 - 31
Ferric hydroxamate binding protein FhuD from Escherichia coli: mutants in conserved and non-conserved regions; Clarke TE et al.; Uptake of iron complexes into the gram-negative bacterial cell requires highly specific outer membrane receptors and specific ATP-dependent (ATP-Binding-Cassette (ABC)) transport systems located in the inner membrane . The latter type of import system is characterized by a periplasmic binding protein (BP), integral membrane proteins, and membrane-associated ATP-hydrolyzing proteins . In gram-positive bacteria lacking the periplasmic space, the binding proteins are lipoproteins tethered to the cytoplasmic membrane . To date, there is little structural information about the components of ABC transport systems involved in iron complex transport . The recently determined structure of the Escherichia coli periplasmic ferric siderophore binding protein FhuD is unique for an ABC transport system (Clarke et al . 2000) . Unlike other BP's, FhuD has two domains connected by a long alpha-helix . The ligand binds in a shallow pocket between the two domains . In vivo and in vitro analysis of single amino acid mutants of FhuD identified several residues that are important for proper functioning of the protein . In this study, the mutated residues were mapped to the protein structure to define special areas and specific amino acid residues in E . coli FhuD that are vital for correct protein function . A number of these important residues were localized in conserved regions according to a multiple sequence alignment of E . coli FhuD with other BP's that transport siderophores, heme, and vitamin B12 . The alignment and structure prediction of these polypeptides indicate that they form a distinct family of periplasmic binding proteins.

J Chemother, 1991 Jan, 3 Suppl 1, 62 - 5
In-vitro activity of cefpodoxime proxetil (RU 51807): a comparative disk diffusion study on isolates from geriatric patients; Ottomano C et al.; 997 strains isolated from clinical specimens arrived at the "Pio Albergo Trivulzio" microbiology laboratory were tested using disks of cefpodoxime, amoxicillin + clavulanic acid, cefaclor, cefuroxime, ofloxacin, cotrimoxazole, ceftriaxone and cefalexin . Gram-positive strains were tested also with erythromycin, while gram-negative bacteria were tested against aztreonam . Cefpodoxime overall activity was well above the effectiveness of the other oral cephalosporins and on the same order as ceftriaxone and ofloxacin . Cefpodoxime proved to be also more active than the combination amoxicillin-clavulanic acid.

J Chemother, 1991 Jan, 3 Suppl 1, 141 - 3
New targets for the mechanism of action of antibiotics active against gram-positive cocci; Canepari P et al.; The main mechanisms of resistance to beta-lactams in gram-positive cocci include either the production of beta-lactamases or alterations in the molecular targets of these antibiotics, the penicillin binding proteins (PBPs) . In spite of the appearance of new beta-lactams, more stable to the hydrolytic activity of beta-lactamases or with higher affinity for PBPs, no substantial progress in improving the activity against gram-positive has been achieved . In addition to the search for new beta-lactams it would be of interest to find molecules directed against new targets of the cell wall of gram-positive bacteria (i.e . teichoic acids and lipoteichoic acid) of which up to now no specific inhibitor is known . These two wall polymers are thought to be essential for cell survival within the host . Among new inhibitors a new antibiotic belonging to the class of acid lipopeptides called daptomycin (LY146032), and active against gram-positive seems of particular interest . Our studies demonstrate that daptomycin is a specific inhibitor of lipoteichoic acid synthesis.

J Biol Chem, 2002 Aug 23, 277(34), 30984 - 90 Epub 2002 May 30.
Unusual mode of binding of human group IIA secreted phospholipase A2 to anionic interfaces as studied by continuous wave and time domain electron paramagnetic resonance spectroscopy; Canaan S et al.; Human group IIA phospholipase A(2) (hGIIA) is secreted from a number of cells during inflammation and is known to interact strongly with anionic membranes and to exhibit potent Gram-positive bactericidal activity . This protein contains 23 cationic residues, which are scattered over its entire surface, resulting in a high pI of 9.39 . To understand the molecular basis for the selective binding of hGIIA to anionic membranes, 14 single-site, spin-labeled hGIIA proteins were analyzed in the presence and absence of vesicles of anionic phospholipid by time domain and continuous wave electron paramagnetic resonance (EPR) spin relaxant techniques . Surprisingly, for hGIIA bound to anionic vesicles, all of the spin labels were highly protected from water-soluble spin relaxants . Together with light scattering studies, these EPR results suggest the formation of a supramolecular aggregate involving clusters of hGIIA molecules bridging together multiple vesicles . This anomalous mode of binding of hGIIA to anionic phospholipid explains previous data in which charge reversal mutation of a few cationic residues on multiple faces of hGIIA leads to a comparable and modest reduction in affinity of the protein for anionic vesicles . In the presence of mixed micelles composed of 10% anionic phospholipids in Triton X-100 a monodisperse protein-lipid complex is formed . Under these conditions, the EPR methods were used to map the surface of hGIIA that constitutes the interfacial binding site (IBS) . The IBS of hGIIA consists of the highly hydrophobic surface that surrounds the opening to the active site slot.

Appl Environ Microbiol, 2002 Jun, 68(6), 2843 - 8
Functional analysis of Sinorhizobium meliloti genes involved in biotin synthesis and transport; Entcheva P et al.; External biotin greatly stimulates bacterial growth and alfalfa root colonization by Sinorhizobium meliloti strain 1021 . Several genes involved in responses to plant-derived biotin have been identified in this bacterium, but no genes required for biotin transport are known, and not all loci required for biotin synthesis have been assigned . Searches of the S . meliloti genome database in combination with complementation tests of Escherichia coli biotin auxotrophs indicate that biotin synthesis probably is limited in S . meliloti 1021 by the poor functioning or complete absence of several key genes . Although several open reading frames with significant similarities to genes required for synthesis of biotin in gram-positive and gram-negative bacteria were found, only bioB, bioF, and bioH were demonstrably functional in complementation tests with known E . coli mutants . No sequence or complementation evidence was found for bioA, bioC, bioD, or bioZ . In contrast to other microorganisms, the S . meliloti bioB and bioF genes are not localized in a biotin synthesis operon, but bioB is cotranscribed with two genes coding for ABC transporter-like proteins, designated here bioM and bioN . Mutations in bioM and bioN eliminated growth on alfalfa roots and reduced bacterial capacity to maintain normal intracellular levels of biotin . Taken together, these data suggest that S . meliloti normally grows on exogenous biotin using bioM and bioN to conserve biotin assimilated from external sources.

J Biotechnol, 2002 Jun 26, 96(2), 129 - 54
Display of proteins on bacteria; Samuelson P et al.; Display of heterologous proteins on the surface of microorganisms, enabled by means of recombinant DNA technology, has become an increasingly used strategy in various applications in microbiology, biotechnology and vaccinology . Gram-negative, Gram-positive bacteria, viruses and phages are all being investigated in such applications . This review will focus on the bacterial display systems and applications . Live bacterial vaccine delivery vehicles are being developed through the surface display of foreign antigens on the bacterial surfaces . In this field, 'second generation' vaccine delivery vehicles are at present being generated by the addition of mucosal targeting signals, through co-display of adhesins, in order to achieve targeting of the live bacteria to immunoreactive sites to thereby increase immune responses . Engineered bacteria are further being evaluated as novel microbial biocatalysts with heterologous enzymes immobilized as surface exposed on the bacterial cell surface . A discussion has started whether bacteria can find use as new types of whole-cell diagnostic devices since single-chain antibodies and other type of tailor-made binding proteins can be displayed on bacteria . Bacteria with increased binding capacity for certain metal ions can be created and potential environmental or biosensor applications for such recombinant bacteria as biosorbents are being discussed . Certain bacteria have also been employed for display of various poly-peptide libraries for use as devices in in vitro selection applications . Through various selection principles, individual clones with desired properties can be selected from such libraries . This article explains the basic principles of the different bacterial display systems, and discusses current uses and possible future trends of these emerging technologies.

Microb Ecol, 2000 Dec, 40(4), 309 - 316
Culture-Independent Detection of Changes in Root-Associated Bacterial Populations of Common Bean (Phaseolus vulgaris L.) Following Nitrogen Depletion; Schallmach E et al.; The structure of root-associated bacterial populations in the legume common bean (Phaseolus vulgaris L.), was studied in plants grown under nitrogen sufficiency and under conditions inducing nitrogen deficiency . Similar cell numbers were obtained in the rhizosphere of nitrogen-amended plants as compared to nitrogen-deficient plants and between various root parts-tip, elongation and branching zones-using DAPI staining . In contrast, a higher proportion of DAPI-stained cells from the nitrogen-amended plants hybridized with a fluorescence-labeled EUB338 probe for the Bacteria domain than cells originating from nitrogen-deficient plants . Shifts in the percentages of EUB338-reactive cells-as well as in absolute cell number-hybridizing to fluorescent rRNA-directed probes specific for the a and g Proteobacteria and for high GC content gram-positive bacteria in separated root segments were detected between the treatments . No such differences were found using b and d Proteobacteria or rRNA group I pseudomonad targeted probes . Denaturating gradient gel electrophoresis profiles of PCR products obtained from the same samples and amplified with Bacteria-domain targeted primers supported the results obtained with the whole cell hybridizations . The advantages and drawbacks of the techniques applied are discussed.

Microb Ecol, 2000 Dec, 40(4), 300 - 308
Structure and Colonization Dynamics of Epiphytic Bacterial Communities and of Selected Component Strains on Tomato (Lycopersicon esculentum) Leaves; Jurkevitch EJ et al.; The sizes and compositions of bacterial populations found on leaves of greenhouse and field grown tomato plants were studied by dilution plating, fatty acid methyl ester analysis (FAME), and BIOLOG plates of isolates in pure cultures . In the greenhouse, overhead-irrigated plants sustained higher microbial populations (up to 105 cfu g-1) than soil-irrigated plants (103 cfu g-1) . Strains isolated from overhead-irrigated plants grown in a vegetable garden (n = 216) and from greenhouse-grown plants (n = 114) were subjected to FAME analysis . Similarly, strains from soil-irrigated field-grown plants (n = 83) were identified using BIOLOG plates . In each case, populations were dominated by a few genera . When concentrated phyllosphere washes (CPW) were sprayed on greenhouse-grown, soil-irrigated plants, leaf bacterial populations of more than 105 CFU g-1 were sustained for 4 days; sterile buffer-sprayed leaves sustained less than 104 CFU g-1 . No significant enrichment of any strain isolated from the sprayed leaves could be detected by FAME identification of randomly selected colonies . However, when recurring leaf saprophytic species (both Gram-positive and Gram-negative) isolated from these experiments and from plants grown outdoors were tested for epiphytic colonization under stressful conditions, all could still be detected at various levels up to 4 days after inoculation, indicating differential epiphytic fitness . The non-epiphytic bacteria Escherichia coli and Azospirillum brasilense disappeared from the leaf surface within the same experimental period.

Microb Ecol, 2001 Jul, 42(1), 11 - 21
The Structure of Microbial Communities in Soil and the Lasting Impact of Cultivation; Buckley DH et al.; The structure of microbial communities was examined as a function of community composition and the relative abundance of specific microbial groups to examine the effects that plant community composition and land-use history have on microbial communities in the soil . The sites sampled were part of the Long Term Ecological Research (LTER) project in agricultural ecology at the W.K . Kellogg Biological Station of Michigan State University (Hickory Corners, MI) and included both active and abandoned agricultural fields as well as nearby fields that had never been cultivated . Microbial community structure was assessed by extracting total RNA from soil samples and using 16S rRNA-targeted oligonucleotide probes to quantify the abundance of rRNA from the alpha, beta, and gamma Proteobacteria, the Actinobacteria (Gram positive bacteria with a high mol % G+C genome), the Bacteria, and the Eukarya . In addition, soil microbial communities were characterized by examining fluorescently tagged terminal restriction fragment length polymorphisms (T-RFLP) in PCR amplified 16S rDNA . Microbial community structure was observed to be remarkably similar among plots that shared a long-term history of agricultural management despite differences in plant community composition and land management that have been maintained on the plots in recent years . In contrast, microbial community structure differed significantly between fields that had never been cultivated and those having a long-term history of cultivation.

Scand J Infect Dis, 2002, 34(3), 206 - 7
Tricuspid valve endocarditis caused by a new species of actinomyces: Actinomyces funkei; Westling K et al.; We present a case of tricuspid valve endocarditis in a 40-y-old woman with a history of heroin abuse . Blood cultures yielded a Gram-positive rod, preliminarily identified as "Actinomyces turicensis-like", but subsequently formally described as Actinomyces funkei sp . nov . The patient was cured by prolonged treatment with 10 weeks of i.v . antibiotics followed by oral antibiotic treatment for 12 weeks.

Science, 2002 May 24, 296(5572), 1459 - 62
Pyrrolysine encoded by UAG in Archaea: charging of a UAG-decoding specialized tRNA; Srinivasan G et al.; Pyrrolysine is a lysine derivative encoded by the UAG codon in methylamine methyltransferase genes of Methanosarcina barkeri . Near a methyltransferase gene cluster is the pylT gene, which encodes an unusual transfer RNA (tRNA) with a CUA anticodon . The adjacent pylS gene encodes a class II aminoacyl-tRNA synthetase that charges the pylT-derived tRNA with lysine but is not closely related to known lysyl-tRNA synthetases . Homologs of pylS and pylT are found in a Gram-positive bacterium . Charging a tRNA(CUA) with lysine is a likely first step in translating UAG amber codons as pyrrolysine in certain methanogens . Our results indicate that pyrrolysine is the 22nd genetically encoded natural amino acid.

Microb Ecol, 2001 Oct, 42(3), 466 - 473
Physiological Diversity of Rhizoplane Diazotrophs of the Saltmeadow Cordgrass, Spartina patens: Implications for Host Specific Ecotypes; Bergholz PW et al.; Diazotrophic bacteria are important contributors to salt marsh productivity, but the biotic and abiotic factors that influence their distributions and function and the extent of their diversity cannot be understood in the absence of physiological information . Here we examine the physiological diversity and distribution patterns of diazotrophic bacteria associated with the rhizoplane of the saltmeadow cordgrass, Spartina patens, in comparison with diazotrophs from other intertidal grasses (tall and short form Spartina alterniflora and Juncus roemerianus) from the same salt marsh . S . patens plants were collected from two distinct habitats, and a total of 115 strains (111 Gram negative and 4 Gram positive strains) were isolated into pure culture by stab inoculating roots and rhizomes into combined nitrogen-free semisolid media . Most strains were microaerophilic and approximately one-half were motile . API test strips were used to eliminate redundancy within the culture collection, resulting in 21 physiologically different API groups (17 Gram negative and 4 Gram positive groups) . A representative strain from each API group was selected for dot blot hybridization with a nifH specific probe and 16 strains (13 Gram negative and 3 Gram positive) were scored as positive . The nifH positive API group representative strains were characterized further using BIOLOG test plates . Substrate utilization potentials defined two S . patens strain clusters, and only one S . patens strain was physiologically similar to any other strain from a different host plant origin . No distinctions could be made based on the different S . patens habitats, suggesting that the host plant may have a greater impact than abiotic environmental conditions on the distributions of the rhizoplane diazotrophs recovered.

J Rheumatol, 2002 May, 29(5), 1093 - 6
Mycobacterium tuberculosis infection of a tophaceous pseudogout nodule; Santos-Ocampo AS et al.; Musculoskeletal infections are uncommon complications of monosodium urate and calcium pyrophosphate dihydrate (CPPD) crystal deposition disease, and frequently involve gram positive and negative organisms . Tumoral calcinosis (tophaceous pseudogout) is a rare manifestation of CPPD deposition disease . We describe a highly unusual case of an infection by Mycobacterium tuberculosis (TB) of a tophaceous pseudogout nodule in a patient with endstage renal disease . The highly destructive nature of this case of combined CPPD arthropathy and musculoskeletal TB underscores the urgency of diagnosing this infection in susceptible patients from countries with high prevalence rates of TB infection.

J Chemother, 2002 Apr, 14(2), 115 - 31
Rokitamycin: bacterial resistance to a 16-membered ring macrolide differs from that to 14- and 15-membered ring macrolides; Braga PC; Rokitamycin is the latest semi-synthetic 16-membered ring macrolide introduced into clinical practice . It is characterized by greater hydrophobicity, better bacterial uptake and a slower release, more cohesive ribosomal binding, and a longer post-antibiotic-effect (PAE) than can be observed with other available 14-, 15- and 16-membered ring macrolides . Rokitamycin exerts its activity on strains that harbor inducible erm genes or the efflux gene, mef(A) . It has also been reported to be more active in vitro than other 16-membered ring macrolides . However, these recognized features are not fully exploited yet because current automated test procedures used in many microbiological laboratories determine susceptibility only to erythromycin or clarithromycin . Resistance to 16-membered ring macrolides cannot be predicted solely on the basis of known resistance to erythromycin or clarithromycin as revealed by an automated susceptibility assay . At least equally important is the knowledge of the bacterial resistance phenotype . This is underlined by the existence of Gram-positive coccal strains resistant to erythromycin and other 14-,15-membered ring macrolides but susceptible to 16-membered ring macrolides . Since the local prevalence of erythromycin phenotypes is generally unknown but might determine the outcome of treatment, the procedure for identifying the phenotypes in erythromycin-resistant strains (which can be easily and cheaply performed using the two- or three-disk assay) should become routine, at least in the countries in which 16-membered ring macrolides are used . This approach should help to optimize the use of macrolides, improve our knowledge of the local prevalence of phenotypes resistant to erythromycin, and offer the possibility of treating infections caused by certain types of erythromycin-resistant pathogens.

Curr Opin Investig Drugs, 2002 Feb, 3(2), 229 - 33
Dalbavancin (Biosearch Italia/Versicor); Steiert M et al.; Biosearch Italia and Versicor are developing dalbavancin, a novel semisynthetic derivative of the natural glycopeptide A-40926 for the potential treatment of Gram-positive infections . In May 2001, the compound entered phase II trials in the US {409929}, {409932} . As of Janaury 2002, phase II trials in skin infections and bacteremia were ongoing {436430} . Both dalbavancin and MDL-63246 are dimethylaminopropyl amide derivatives of A-40926; dalbavancin differs from MDL-63246 in its acylamino sugar, which consists of glucuronic acid in dalbavancin and glucosamine in MDL-63246 {216093}, {298527} . The development of MDL-63246 has been discontinued {298527} . In January 1999, Versicor gained North American market rights to BI-397 {311500} . In January 2002, UBS Warburg predicted filing of an NDA in the second half of 2003 for dalbavancin, for treatment of complicated skin and soft tissue infections {439472} . In December 2001, Lehman Brothers predicted a late 2005 launch for dalbavancin, with sales of US $7.7 million in 2005, US $65.8 million in 2006, and peak sales of US $225 million in 2008 {439469}.

Dermatology, 2002, 204 Suppl 1, 37 - 41
Evaluation of the bactericidal activity of povidone-iodine and commercially available gargle preparations; Shiraishi T et al.; OBJECTIVE: Encouragement of gargling is important for the control of opportunistic and community-acquired infections . In hospitals, a povidone-iodine (PVP-I) gargle is used frequently . However, at pharmacies in the community a variety of gargles containing various ingredients are now available . In view of this, we conducted a study to compare the bactericidal activities of a PVP-I gargle with those of other commercially available gargles . In addition, we asked about the feeling after use by questionnaire . At middle schools in our city, we investigated whether the encouragement to use the PVP-I gargle had an effect on the absence rate from school due to common cold and influenza . METHODS: In vitro, using 3 strains of gram-positive and 4 strains of gram-negative bacteria as the test strains, the bactericidal activities of the PVP-I, chlorhexidine gluconate (CHG) and cetylpiridium chloride gargles (CPC) were compared by the contact test method . In vivo, with subjects in groups of 6 each, the reduction rate in the oral bacterial count after gargling as compared to the baseline count before gargling was determined and compared among the 3 gargling agents used . In addition, a questionnaire study was conducted to compare the feeling after use of the 3 gargling agents . Whether the absence rate due to common cold and influenza changed by encouraging the use of the PVP-I gargle was determined by comparing a middle school where the PVP-I gargle was used and other middle schools where it was not . RESULTS: (1) PVP-I killed all the test strains after 30 s of exposure . (2) The mean reduction rate in bacterial count immediately after gargling was 99.4% for PVP-I, 59.7% for CHG and 97.0% for CPC . (3) Findings of the questionnaire study revealed that the PVP-I gargle was evaluated highest in terms of taste, feeling after gargling and odor among all the gargles tested . (4) At the middle school where the use of the PVP-I gargle was encouraged, the absence rate due to common cold and influenza was significantly lower as compared to those at middle schools where another gargle was used . CONCLUSION: Of the 3 gargles tested, PVP-I showed the highest bactericidal rate and the highest reduction rate in oral bacterial count . Encouragement of the use of the PVP-I gargle contributed to the decrease in absence rates due to common cold and influenza, indicating that encouragement of gargling with PVP-I is useful for the prevention of common cold and influenza .

Braz J Infect Dis, 2001 Dec, 5(6), 356 - 9
Bilateral endogenous endophthalmitis associated with infective endocarditis: case report; Arcieri ES et al.; Bilateral endogenous endophthalmitis is a rare condition initiated by infection by microbes in the bloodstream, such as those arising from a foci of infective endocarditis . We report a case and discuss the diagnostic aspects and the clinical outcome of a patient with characteristic findings of the disease . The patient was a 49 year old white male who had a metallic aortic valve implanted 7 months previously, and who presented to the hospital with 10 days of fever, cough and dyspnea, then diarrhea and mental confusion . On the second day of hospitalization, he experienced sudden loss of vision in both eyes . A Gram-positive coccobacillus was isolated from the bloodstream, he was treated with fluoroquinolone with disappearance of fever, decreased ocular inflammation, and improvement in his vision to light perception . He later underwent valve replacement surgery but died during the procedure . We review the occurrence of ocular signs and symptoms and their importance in patients with endocarditis.

FEMS Microbiol Lett, 2002 Apr 9, 209(2), 141 - 8
Carbon catabolite repression in bacteria: choice of the carbon source and autoregulatory limitation of sugar utilization; Bruckner R et al.; Carbon catabolite repression (CCR) in bacteria is generally regarded as a regulatory mechanism to ensure sequential utilization of carbohydrates . Selection of the carbon sources is mainly made at the level of carbohydrate-specific induction . Since virtually all carbohydrate catabolic genes or operons are regulated by specific control proteins and require inducers for high level expression, direct control of the activity of regulators or control of inducer formation is an efficient measure to keep them silent . By these mechanisms, bacteria are able to establish a hierarchy of sugar utilization . In addition to the control of induction processes by CCR, bacteria have developed global transcriptional regulation circuits, in which pleiotropic regulators are activated . These global control proteins, the catabolite gene activator protein (CAP), also known as cAMP receptor protein, in Escherichia coli or the catabolite control protein (CcpA) in Gram-positive bacteria with low GC content, act upon a large number of catabolic genes/operons . Since practically any carbon source is able to trigger global transcriptional control, expression of sugar utilization genes is restricted even in the sole presence of their cognate substrates . Consequently, CAP- or CcpA-dependent catabolite repression serves as an autoregulatory device to keep sugar utilization at a certain level rather than to establish preferential utilization of certain carbon sources . Together with other autoregulatory mechanisms that are not acting at the gene expression level, CCR helps bacteria to adjust sugar utilization to their metabolic capacities . Therefore, catabolic/metabolic balance would perhaps better describe the physiological role of this regulatory network than the term catabolite repression.

Pediatr Infect Dis J, 2002 Mar, 21(3), 227 - 34
Infections diagnosed in the first year after pediatric stem cell transplantation; Benjamin DK Jr et al.; BACKGROUND: Cumulative incidence of infections in the first year posttransplantation in adult patients has been well-described . Such description is less than complete for pediatric stem cell transplantation (SCT) patients . Further among those patients who have been infected, analysis of risk factors for infection has not been well-described for a large cohort of pediatric SCT patients . METHODS: We conducted a retrospective cohort study of infections in the first year after SCT at Duke University Medical Center . We recorded all infections in the first year after transplantation . We determined incidences for 6 categories of infection: gram-negative rods; gram-positive cocci; yeast species; Aspergillus sp.; adenovirus; and cytomegalovirus . We determined incidences based on type of transplant and days post transplantation . We also completed bivariable and multivariable analysis of risk factors {neutropenia, graft vs . host disease (GVHD) and GVHD treatment} for infection type among those children who were infected . RESULTS: We evaluated 510 transplants in 485 children . There were 584 infections in the first year after transplantation . During the first 30 days posttransplantation, type of transplantation did not predict incidence of infection or type of infection . After 30 days children who received unrelated cord blood transplant and matched unrelated donor transplant were at much higher risk of infection than were patients who received autologous, matched sibling or haploidentical transplant (P < 0.001) . Patients who received unrelated cord blood or matched unrelated donor transplantation were at higher risk of aspergillosis (P = 0.002), candidiasis (P = 0.005) and adenovirus (P < 0.0001) but not cytomegalovirus (P = 0.18) . In analysis of risk factors among those infected, patients with aspergillosis were more likely to have severe GVHD: multivariable 1 year risk ratio, 7.5; 95% confidence interval, 3.0, 18.4 . Neutropenia was more strongly associated with gram-negative rod infection than any other type of infection . CONCLUSIONS: The incidence of infection immediately after transplantation did not differ significantly by type of transplant in this pediatric population . Type of transplant predicted increased incidence of infection 30 days posttransplantation and increased incidence of infection with several organisms traditionally associated with a high mortality rate in the transplant population.

Clin Diagn Lab Immunol, 2002 May, 9(3), 530 - 43
Endotoxin-induced gamma interferon production: contributing cell types and key regulatory factors; Varma TK et al.; Gamma interferon (IFN-gamma) is an important mediator of endotoxin (lipopolysaccharide {LPS})-induced immune responses . However, the specific cell types that produce IFN-gamma in response to LPS and the cellular factors that regulate LPS-induced IFN-gamma production have not been fully determined . The present studies were undertaken to characterize the cell populations that produce IFN-gamma after LPS challenge in the spleens of mice and to determine the regulatory factors that modulate LPS-induced production of IFN-gamma . Our studies show that the levels of splenic IFN-gamma mRNA and protein production peak at 6 and 8 h, respectively, after systemic LPS challenge . Approximately 60% of IFN-gamma-producing cells are natural killer (NK) cells (CD3(-)DX5(+)) and 25% are NKT cells (CD3(+)DX5(+)) . Most of the remaining IFN-gamma-producing cells are T cells (CD3(+)DX5(-)), macrophages, and dendritic cells . Functionally, interleukin-12 (IL-12) is the major IFN-gamma-stimulating factor after LPS challenge, with costimulation provided by IL-15, IL-18, and B7 proteins . IL-10 is a major inhibitor of LPS-induced IFN-gamma production . Unlike intact heat-killed gram-negative and gram-positive bacteria, the class II major histocompatibility complex did not play a functional role in LPS-induced IFN-gamma production . LPS is a potent stimulus for splenic IL-10, IL-12 p40, and IL-15 mRNA expression, whereas IL-12 p35 and IL-18 mRNAs, as well as B7 proteins, are constitutively expressed in the mouse spleen . Of the factors studied, IL-18 serves as the most potent costimulus with IL-12 for IFN-gamma production, followed by IL-15 and B7 proteins . These data demonstrate that NK cells and NKT cells are the most abundant IFN-gamma-producing cells in the mouse spleen after LPS challenge and that IL-10 and IL-12 are key functional regulators of LPS-induced IFN-gamma production.

Przegl Epidemiol, 2001, 55 Suppl 3, 41 - 5
{Monitoring of bacteremia in oncological patients}; Osmanska M et al.; The aim of the present study was analysis of bacteremia occurring among oncological patients treated in 3 departments of Regional Center of Oncology, in period 1997-2000 . A total number of 255 blood cultures from 89 patients were tested using the automatic system to early detection of positive blood-cultures Bactec 9050 (Becton Dickinson) . The strains were identified in the automatic VITEK system using commercial strips with biochemical tests and in manual system API (bioMerieux) . The total number of positive blood cultures was 70 (27.45%) . The most frequently isolated causal agents were the Gram-positive microorganisms (65.79%) . Among 28 examined patients with positive blood cultures 10 were with lymphoma and 9 with cancer of the gastrointestinal tract . 9 patients (32.14%) had sepsis, 4 patients with sepsis died . Constant monitoring of bacteremia in oncological patients should be fundamental element in control of hospital infection program.

Microb Ecol, 2002 Jan, 43(1), 82 - 91 Epub 2002 Jan 23.
Bacterial distribution and phylogenetic diversity in the Changjiang estuary before the construction of the Three Gorges Dam; Sekiguchi H et al.; The bacterial community structure in the Changjiang estuary was studied for comparison with future changes, related to the construction of the Three Gorges Dam . Population densities of bacteria in the surface water at station C1 estimated by CFU on marine agar plates and by DAPI direct count, were 2.8 x 10(4) ml(-1) and 4.2 x 10(5) ml(-1), respectively . Physicochemical properties of water, such as temperature and salinity, suggested that station C1 was affected by freshwater from the Changjiang River . Cluster analysis of the PCR-RFLP patterns obtained from 9 samples showed that the bacterial community structure at station C1 was different from the structure at the other stations . Bacterial diversity in the surface water at station C1 was studied based on the genotypes of the 250 clones of 16S rRNA, and on the phenotypes generated on Biolog GN plates for 70 isolates . Sequences of bacteria from two common marine groups, alpha- and gamma-Proteobacteria, were frequently observed . Some other divisions, including the beta-Proteobacteria, C/F/B group, low G+C gram positive, high G+C gram positive, chloroplasts, and relatives of Verrucomicrobia were also observed . The putative dominant species based on both genotype and phenotype analyses were close relatives of Alteromonas macleodii or Roseobacter spp . These results reflected the nutrient-rich environment at station C1.

Crit Care, 2002 Apr, 6(2), 149 - 54 Epub 2002 Mar 06.
Complicated infective endocarditis necessitating ICU admission: clinical course and prognosis; Karth G et al.; AIM: To study incidence, clinical course and prognostic factors in patients admitted to medical intensive care units (ICUs) because of a complicated course of infective endocarditis . METHOD: This was a retrospective multicenter observational study of 4106 patients admitted to four medical ICUs in one tertiary hospital and one university hospital between 1994 and 1999 . RESULTS: Infective endocarditis was identified in 33 (0.8%) patients . Of these, 26 were male, mean age was 59 +/- 12 and APACHE-III score was 75 +/- 31 . Reasons for transfer to the ICU were congestive heart failure in 64%, septic shock in 21%, neurological deterioration in 15% and cardiopulmonary resuscitation in 9% . Inotropes or vasoconstrictors were required in 73% and multiorgan failure developed in 64% of the patients . Prosthetic valve endocarditis was present in 21% . Gram-positive cocci were found in 96% of all positive cultures; cultures were negative in 27% of the patients . Transthoracic echocardiograms were diagnostic in only 33% and transesophageal studies were required in 91% to confirm diagnosis or fully to delineate the extent of disease . Surgical intervention was performed in 60% of the patients, and the remaining 40% were only treated medically . The APACHE-III score on admission did not differ statistically between the two groups (69 +/- 30 versus 84 +/- 34, P = 0.17) . In-patient mortality was 84% in patients treated medically, and 35% in surgically treated patients . Using multivariate analysis, acute renal failure on admission was identified as the independent single predictor for in-patient death (OR 5, 95% CI 1.04-24.03, P = 0.04) . CONCLUSION: The prognosis for patients with infective endocarditis requiring admission to a medical ICU is serious . Nevertheless, the data suggest that surgical intervention may be successfully performed in a substantial number of patients despite the presence of severe shock and occurrence of multiorgan failure.

Transpl Int, 2002 Apr, 15(4), 163 - 72 Epub 2002 Mar 21.
Lipopolysaccharide-binding protein as a new and reliable infection marker after kidney transplantation; Kaden J et al.; The early and reliable differentiation of rejections, viral infections and bacterial infections is one of the main problems after organ transplantation . One promising solution to this problem is the lipopolysaccharide-binding protein (LBP), which is regulated upwards in gram-negative sepsis and related conditions . Therefore, the aim of our study was to explore the diagnostic potential of LBP serum levels in well-defined, non-infectious and infectious events after kidney transplantation (KTx) . In a retrospective study the LBP serum levels were measured in a total of 686 serum samples from 52 kidney graft recipients . In all pre-KTx sera tested, the mean LBP level was 8.8+/-3.5 microg/ml (reference range: 2.0-15.2 microg/ml) . In 7 of 52 recipients without intraoperative T-cell depletion, the mean LBP level was significantly ( P<0.01) increased (13.0+/-1.5 microg/ml) at post-KTx day 1, but was within the reference range . In contrast, the intraoperative T-cell depletion by antilymphocyte antibodies resulted in a significant ( P<0.01) increase to 25.8+/-11.4 microg/ml (range: 13.3-47.2 microg/ml) . In recipients with immediate ( n=14) or delayed ( n=9) graft function without any other complications, all post-KTx values (except the post-KTx peak) were within the reference range . In 10 recipients with steroid-sensitive rejections and in 11 recipients with steroid-resistant rejections, no rejection-associated changes of the LBP levels could be shown . In six recipients with cytomegalovirus infection, the detection of an antigenemia (pp65) also was not associated with alterations of the LBP levels . In addition, there was no correlation between LBP levels and the number of pp65-positive leukocytes in peripheral blood . In contrast, a strong elevation of LBP levels was seen in five recipients with gram-positive bacteremia as well as in other severe bacterial infections (e.g., purulent extravasate, heavily infected grafts, bacterial pneumonia and contaminated hematoma) . In two recipients with superinfected (bacterial and mycotic or viral) Pneumocystis carinii pneumonias requiring assisted ventilation, LBP levels were elevated, too . Thus, in our study only systemic non-viral infections and massive lymphocytolysis were associated with elevated LBP serum levels.

Trends Microbiol, 2002 May, 10(5), 205 - 8
Anchorless adhesins and invasins of Gram-positive bacteria: a new class of virulence factors; Chhatwal GS; Bacterial adherence to and invasion of eukaryotic cells are important mechanisms of pathogenicity . Most Gram-positive bacteria interact with the components of the host extracellular matrix (ECM) to adhere to, colonize and invade cells and tissues . The bacterial proteins that bind to components of the ECM harbour signal sequences for their secretion and mechanisms of anchoring to the host cell surface . However, in recent years, some cell-surface adhesins and invasins of Gram-positive bacteria have been described that do not possess a signal sequence or a membrane anchor . These proteins are secreted by an as-yet-unknown mechanism and are probably localized on the bacterial surface by reassociation . These anchorless but surface-located adhesins and invasins represent a new class of virulence factors.

Immunology, 2002 May, 106(1), 38 - 45
Involvement of ERK, p38 and NF-kappaB signal transduction in regulation of TLR2, TLR4 and TLR9 gene expression induced by lipopolysaccharide in mouse dendritic cells; An H et al.; Toll-like receptors (TLR) are sentinel receptors capable of recognizing pathogen-associated molecule patterns (PAMP) such as lipopolysaccharide (LPS) and CpG-containing oligonucleotides (CpG ODN) . TLR2 and TLR4 are major receptors for Gram-positive and Gram-negative bacterial cell wall components, respectively . TLR9 is necessary for CpG signalling . LPS or CpG ODN can activate immature dendritic cells (DC) and induce DC maturation characterized by production of cytokines, up-regulation of co-stimulatory molecules, and increased ability to activate T cells . However, little is known regarding the regulation of TLR gene expression in mouse DC . In this study, we investigated the regulation of TLR2, TLR4 and TLR9 gene expression by LPS in murine immature DC . TLR2, TLR4 and TLR9 mRNA were up-regulated following LPS stimulation . The up-regulation of TLR9 expression coincided with significantly increased production of tumour necrosis factor-alpha induced by LPS plus CpG ODN . While inhibition of extracellular signal-related kinase and NF-kappaB activation suppressed the up-regulation of the expression of TLR2, TLR4 and TLR9 mRNA, inhibition of p38 kinase prevented the up-regulation of TLR2 and TLR4 mRNA expression but enhanced the up-regulation of TLR9 expression . These results demonstrated that TLR2, TLR4 and TLR9 gene expression was differently regulated by LPS in mouse immature DC . Up-regulation of TLR2, TLR4 and TLR9 expression by LPS might promote the overall responses of DC to bacteria and help to explain the synergy between LPS and other bacterial products in the induction of cytokine production.

Mol Biol (Mosk), 2002 Mar-Apr, 36(2), 228 - 39
{Bacterial genomic islands: organization, function, and role in evolution}; Il'ina TS et al.; Data on the structural organization and evolutionary role of specific bacterial DNA regions known as genomic islands are reviewed . Emphasis is placed on the most extensively studied genomic islands, pathogenicity islands (PAIs), which are present in the chromosome of Gram-negative and Gram-positive pathogenic bacteria and absent from related nonpathogenic strains . PAIs are extended DNA regions that harbor virulence genes and often differ in GC content from the remainder of the bacterial genome . Many PAI occur in the tRNA genes, which provide a convenient target for foreign gene insertion . Some PAI are highly homologous to each other and contain sequences similar to ISs, phage att sites, and plasmid ori sites, along with functional or defective integrase and transposase genes, suggesting horizontal transfer of PAI among bacteria.

Mol Biol (Mosk), 2002 Mar-Apr, 36(2), 216 - 27
{Horizontal transfer of mercury resistance genes in natural bacterial populations}; Mindlin SZ et al.; The results of studying the horizontal transfer of mercury resistance determinants in environmental bacterial populations are reviewed . Identical or highly homologous mercury resistance (mer) operons and transposons were found in bacteria of different taxonomic groups from geographically distant regions . Recombinant mer operons and transposons were revealed . The data suggest high frequencies of horizontal transfer and of recombination for mercury resistance determinants . The mechanisms of horizontal gene transfer were elucidated in Gram-negative and Gram-positive bacteria . New transposons were found and analyzed.

Mol Microbiol, 2002 Apr, 44(1), 195 - 204
Methylation at nucleotide G745 or G748 in 23S rRNA distinguishes Gram-negative from Gram-positive bacteria; Liu M et al.; Bacteria tune the function of their ribosomes by methylating specific rRNA nucleotides . Nucleotide G745 in Escherichia coli 23S rRNA is methylated by the methyltransferase enzyme RrmA, whereas in Streptomyces fradiae, the neighbouring nucleotide G748 is methylated by the enzyme TlrB . Both nucleotides line the peptide exit channel of the ribosome at the binding site of macrolide, lincosamide and streptogramin B antibiotics . Resistance to the macrolide tylosin, which is produced by S . fradiae, is conferred by methylation of G748 . RrmA and TlrB are homologues (29% identical), and a database search against all presently available sequences revealed a further two dozen homologues from a wide variety of Bacteria . No homologues were found among the Archaea or Eukarya . The bacterial sequences adhere to the species phylogeny and segregate into two groups, in which the Gram-negative sequences align with RrmA and the Gram-positives with TlrB . Consistently, in more than 20 species tested, the distribution of methylation in the Gram-negative rRNAs (methylated at G745) and the Gram-positives (methylated at G748) perfectly matches the bacterial phylogeny . Cloning and expression of representative methyltransferase genes showed that this specificity of methylation is determined solely by the methyltransferase enzyme and is independent of the origin of the rRNA substrate . This is the first case in which the position of an RNA methylation defines a sharp division between the Gram-negative and Gram-positive bacteria . Given the specificities and distribution of these methyltransferases, we propose a change in the nomenclature of RrmA to RlmAI (rRNA large subunit methyltransferase) and of TlrB to RlmAII.

Eur J Biochem, 2002 Apr, 269(7), 1827 - 34
Binding of hemolin to bacterial lipopolysaccharide and lipoteichoic acid . An immunoglobulin superfamily member from insects as a pattern-recognition receptor; Yu XQ et al.; Hemolin, a plasma protein from lepidopteran insects, is composed of four immunoglobulin domains . Its synthesis is induced by microbial challenge . We investigated the biological functions of hemolin in Manduca sexta . It was found to bind to the surface of bacteria and yeast, and caused these micro-organisms to aggregate . Hemolin was demonstrated to bind to lipopolysaccharide (LPS) from Gram-negative bacteria and to lipoteichoic acid from Gram-positive bacteria . Binding of hemolin to smooth-type forms of LPS was competed for efficiently by lipoteichoic acid and by rough mutant (Ra and Rc) forms of LPS, which differ in polysaccharide length . Binding of hemolin to LPS was partially inhibited by calcium and phosphate . Hemolin bound to the lipid A component of LPS, and this binding was completely blocked by free phosphate . Our results suggest that hemolin has two binding sites for LPS, one that interacts with the phosphate groups of lipid A and one that interacts with the O-specific antigen and the outer-core carbohydrates of LPS . The binding properties of M . sexta hemolin suggest that it functions as a pattern-recognition protein with broad specificity in the defense against micro-organisms.

Zh Mikrobiol Epidemiol Immunobiol, 2002 Jan-Feb, (1), 70 - 1
{Etiology and structure of infectious complications of cytostatic therapy in children with acute lymphoblastic leukemia and non-B-cell non-Hodgkin lymphomas}; Peshikova MV et al.; A retrospective analysis of medical histories with acute lymphoblast leucosis and non-B-cell non-Hodgkin lymphomas made it possible to reveal infectious complications of cytostatic therapy in 100% of children, namely: sepsis (0.3%), unidentified infection (12%), local infection (87.7%) . Infectious complications of the cytopenic nature were localized mainly in the upper sections of the gastrointestinal tract and in upper respiratory tract . Bacterial infectious complications caused by opportunistic microorganisms with the prevalence of Gram positive flora, resistant to cephalosporins of generations I and II, occurred most frequently.

Anal Biochem, 2002 Apr 15, 303(2), 176 - 85
Carbohydrate analysis of bacterial polysaccharides by high-pH anion-exchange chromatography and online polarimetric determination of absolute configuration; Stroop CJ et al.; A significant problem in structure determination of complex carbohydrates, especially for bacterial polysaccharides, is determination of the absolute configuration of the component monosaccharides . A number of analytical methods have been used for this purpose but, as a result of the wide variety of chemical properties of sugars found in complex polysaccharides, no single method is universally applicable . High-resolution gas chromatography of volatile derivatives with chiral reagents is the most widely used method . Optical activity, although direct and simple, lacks sensitivity generally requiring a large quantity of pure monosaccharide . We report a combination of high-performance anion-exchange chromatography (HPAEC) with combined electrochemical pulsed amperometric detection and in-line detection of optical rotation with an in-line laser polarimeter for analysis of a number of sugars found in complex polysaccharides . We show that application of the method for analysis of capsular polysaccharides of several gram-positive and gram-negative pathogenic bacteria provides useful information simultaneously on carbohydrate composition and the enantiomeric configuration of component sugars . (C)2002 Elsevier Science (USA)

Curr Pharm Des, 2002, 8(9), 815 - 33
Multiple activities in lantibiotics--models for the design of novel antibiotics?
Pag U, Sahl HG.
Lantibiotics are antibiotic peptides distinguished by the presence of the rare thioether amino acids lanthionine and/or methyllanthionine . They are produced by Gram-positive bacteria as gene-encoded precursor peptides and undergo post-translational modification to generate the mature peptide . The structural gene for the prepeptide and the genes involved in biosynthesis, processing, export as well as regulation and producer strain self-protection are organized in clusters . Based on their structural and functional features lantibiotics are currently divided into two major groups . The flexible amphiphilic type-A lantibiotics act primarily by pore formation in the bacterial membrane, a mechanism which was recently shown, e.g . for nisin and epidermin, to involve the interaction with specific docking molecules such as the membrane precursor lipid II . The rather rigid and globular type-B lantibiotics inhibit enzyme functions through interaction with the respective substrates: mersacidin and actagardine inhibit the cell wall biosynthesis by complexing lipid II, whereas the cinnamycin-like peptides inhibit phospholipases by binding phosphoethanolamine . Lantibiotics have attracted much attention in recent years and undergone extensive characterization . New insights into the mode of action and structure-function relationships as well as the biochemistry and the genetics will be outlined in this review.

Biochem Biophys Res Commun, 2002 Apr 12, 292(4), 880 - 5
Identification of LPS-binding peptide fragment of MD-2, a toll-receptor accessory protein; Mancek M et al.; Members of the toll-like receptor family are crucial in recognition of microbial pathogens as part of innate immune response . MD-2, an accessory protein to TLR4, present on the extracellular side of the membrane is needed to initiate the signal transduction . We have identified a 15 amino acid region of human MD-2 that contains several features of other lipopolysaccharide (LPS) binding proteins and peptides . In vitro LPS neutralization by this peptide was observed and confirmed by 2D transferred NOESY NMR experiments . NMR experiments have also shown binding of the MD-2 peptide to lipoteichoic acid (LTA) but not to peptidoglycan . Furthermore this peptide inhibited growth of gram-negative and to a lower extent of some gram-positive bacteria . Our results indicate that this region of MD-2 might be responsible for binding of LPS and confirms the role of MD-2 as an accessory protein in LPS signaling bestowing the Toll receptors their specificity . (c)2002 Elsevier Science (USA).

Cell Mol Biol Lett, 2002, 7(1), 61 - 7
Fatty acid profiling: its usefulness in the evaluation of microbial associations with the green microalga Apatococcus constipatus; Zarnowski R; To determine differences in microbial community structures, fatty acids from two strains of the green microalga Apatococcus constipatus were isolated and identified by instrumental means . The main fatty acids found were 16:0 and 14:0 . These predominant acids represented more than 53% of the total fatty acid (content in both algal isolates . In addition, saturated fatty acids were present in much greater quantity than unsaturated ones . Differences between the strains in the composition of other, modified fatty acids were also evident . The occurrence of fatty acid biomarkers characteristic of certain taxonomic groups confirmed the presence of Gram-positive and Gram-negative bacteria, and fungi . Those observed variations were undoubtedly due to distinct community structures of symbiotic microorganisms living in close associations with the alga . The results presented here indicate that different isolates of the same alga might exhibit different microbial community structures.

Hunan Yi Ke Da Xue Xue Bao, 1999, 24(2), 136 - 8
{PCR for the detection of 16S rRNA gene bacteria}; Li C et al.; According to the high conservative region of 16S rRNA gene in bacteria, PCR primers of the broad-range bacteria, gram-positive bacteria and gram-negative bacteria were synthesized to detect 13 bacterium species and (40) clinical specimens . All the tested bacterium species were positive . The lowest concentration of Escherichia Coli detected by serial dilution was 4 CFU.ml-1 . The positive rate of PCR (27/40) was higher than that of bacterium culture(18/40) . The results indicate that these PCR primers possess high specificity and sensitivity in identifying 16S rRNA gene of bacteria.

Pediatr Med Chir, 2002 Jan-Feb, 24(1), 13 - 20
{Nosocomial infections in pediatric and neonatal intensive care: an epidemiological update.}; Fanos V et al.; Hospital-acquired infection (nosocomial infection) is a world-wide problem . The peculiar susceptibility of newborns and children, the widespread use of antibiotics, advances in hospital practice, have resulted in an increased incidence and recognition of neonatal and pediatric bacterial nosocomial infections . The aim of this review is to present an update on epidemiology of hospital-acquired infections, quoting recent and relevant papers on this topic . Main risk factors for the neonatal and the pediatric intensive care infections are presented, respectively . Emerging antibiotic-resistant gram-positive and gram-negative bacteria in intensive care units are also considered . Infection control programmes are necessary to monitor and to prevent nosocomial infections.

J Chemother, 2001 Nov, 13 Spec No 1(1), 173 - 80
Pathogenesis of septic shock: implications for prevention and treatment; Calandra T; The innate immune system is in the vanguard of host defenses against infection . Recognition of invasive microbial pathogens is mediated by pattern recognition receptors on the surface of immune cells that recognize pathogen-associated molecular motifs . Considerable progress has been made in recent years in understanding how bacterial products initiate sepsis . In gram-negative sepsis, the LPS-binding protein (LBP), CD14 and the recently identified Toll-like receptor 4 (TLR4) are key molecules for the recognition of endotoxin (lipopolysaccharide, LPS) by cells of the myelomonocytic lineage . In gram-positive sepsis, components of the bacterial cell wall (peptidoglycan, PGN; lipoteichoic acids, LTA) have been shown to activate myeloid cells through an interaction with a receptor complex composed of CD14, TLR2 and perhaps also TLR6 (PGN) or CD14 and TLR4 (LTA) . By contrast, gram-positive exotoxins act as superantigens and directly stimulate T lymphocytes by cross-linking the MHC class II of antigen presenting cells to specific chains of the T cell receptor . Immune cells activated by microbial pathogens release numerous effector molecules, which orchestrate the innate and adaptive host defenses . Furthermore, bacteria and microbial toxins directly activate the complement and coagulation systems, which play an important part in the host defensive response . Severe sepsis and septic shock can be viewed as clinical manifestations of a failing innate immune response that ultimately results in an overstimulation of the physiological host response . The pathogenesis of sepsis is far more complex that was initially anticipated . However, combined research efforts of basic scientists and clinical investigators continue to provide critical information for the identification of novel therapeutic targets . The exciting results obtained recently with treatment strategies designed to correct coagulation abnormalities occurring during sepsis are an example of how research may ultimately translate into improved patient care.

J Chemother, 2001 Nov, 13 Spec No 1(1), 159 - 72
Antibiotic induced endotoxin release and clinical sepsis: a review; Holzheimer RG; Sepsis and peritonitis have not lost much of their danger for patients . The mortality rate in peritonitis has only marginally decreased during the last 30 years despite aggressive surgical and sophisticated intensive care treatment . In intra-abdominal infection and peritonitis source control remains the mainstay of treatment, although general principles and denominators of successful source control need to be established . Endotoxin has been recognized as a major player in the pathogenesis of sepsis and its significance in clinical disease has been investigated in clinical studies for more than 20 years . Since the Sixties there is a growing interest in the effect of antibiotics and other compounds on the release of endotoxin . The effect of antibiotics on the release of endotoxin and inflammatory parameters, e.g., cytokines, remains to be clarified despite a growing body of in-vitro studies, animal studies and a few clinical studies . The purpose of this review is to evaluate the evidence of endotoxin release in clinical studies and the effect that antibiotic treatment may have in-vitro, in-vivo and in clinical studies on endotoxin and cytokine release . In-vitro antibiotic-induced endotoxin release may depend on antibiotic class, presence of serum, type of organism, site of antibiotic action and Gram-stain . Endotoxin release may be different in late or early lysis, proportional to the number of killed pathogens . Morphology of bacteria may have an impact on endotoxin release and phagocytosis . Antibiotic-treated animals may show higher endotoxin levels with a higher survival rate than untreated animals . Plasma endotoxin may increase despite decreasing bacteremia . There may be a similar killing rate by different antibiotics but a difference in endotoxin release . Intestinal endotoxin does not necessarily correlate to the level of gram-negative bacteria . However, the alteration of the gut content by pretreatment may be associated with reduced endotoxemia and increased survival . Antibiotic-induced endotoxin release may be different depending on the type of infection, the location of infection, the virulence of strains, Gram-stain, mode of application and dosage of antibiotic . Different antibiotics may induce the release of different forms of endotoxin which may be lethal for sensitized animals . The combination of antibiotics with inhibitors of endotoxin or the pro-inflammatory response may be responsible for increased survival by decrease of endotoxin release . The clinical significance of antibiotic-induced endotoxin release is documented only in a few clinical disorders, e.g., meningitis, urosepsis . The difference in endotoxin release by PBP 2-specific antibiotics, e.g., imipenem, and PBP 3-specific antibiotics, e.g., ceftazidime, may not be visible in each study . Patients with increased multi-organ failure (MOF) scores may profit from treatment with antibiotics known to decrease endotoxin . In conclusion, the clinical significance of antibiotic-induced endotoxin release remains to be clarified . Type of pathogen and its virulence may be more important than recently suggested . gram-positive pathogens were just recently recognized as an important factor for the development of the host response . In case of fever of unknown origin in intensive care patients either failure of treatment, e.g., failure of source control in intra-abdominal infection, or a side effect of antibiotic treatment, e.g., endotoxin release, should be considered as a cause of the fever.

J Chemother, 2001 Nov, 13 Spec No 1(1), 153 - 8
Mechanisms of tissue injury in sepsis: contrasts between gram positive and gram negative infection; Cohen J; Sepsis is the systemic response to infection, and is caused by gram positive and gram negative bacteria in approximately equal numbers . In recent years it has become increasingly clear that there are significant differences between sepsis caused by gram positive and gram negative infections, both in the microbial components that initiate the injury, and the host responses that follow it . These differences will assume greater importance in developing targeted therapeutic interventions to severe sepsis.

J Chemother, 2001 Nov, 13 Spec No 1(1), 123 - 8
Antibiotic prophylaxis in clean surgery: vascular surgery; Pratesi C et al.; The commonest and most dangerous infective complication in vascular surgery is prosthetic graft infection . Despite the use of routine systemic antibiotic prophylaxis, graft infection occurs after 3-5% of all prosthetic vascular reconstructions . Infection is associated with a high rate of major morbidity and mortality, with significant time and cost implications . Management can be difficult and the outcome is often disappointing . Data from Italian Registry of Prosthetic Graft Infections show that the commonest site of infection is the aorto-femoral district and that involved bacteria are usually gram-positive in early and low-grade infections and gram-negative in late and high-grade infections . Results are poor, with a mortality rate of 15% for elective surgery and of 60% for emergency interventions . We report the results of a multicenter randomized controlled trial of rifampicin-bonded Dacron grafts in aorto-femoral surgery, in which our Department was involved . Data demonstrate a reduction in total early wound and graft infection rates, and 2-year results show a small, non-significant reduction in graft infection (1.7% in study group, 2.3% in control group) . The same results were obtained in two other multicenter trials.

Pigment Cell Res, 2002 Apr, 15(2), 104 - 11
Cloning and molecular characterization of a SDS-activated tyrosinase from Marinomonas mediterranea; Lopez-Serrano D et al.; The sequence of the tyrosinase gene cloned from Marinomonas mediterranea is reported . It is the second tyrosinase cloned from a Gram negative bacterium . Its size is higher than that of Gram positive tyrosinases from Streptomyces, and more similar to the eukaryotic enzymes . Its sequence shares the features of copper-binding sites found in all tyrosinases . Based in the comparison of tyrosinases from all types of organisms, an extension of the characteristic signatures existing at Prosite is proposed . This tyrosinase shares with some plant and amphibian tyrosinases a strong specific activation by submicellar concentrations of SDS . Intrinsic fluorescence and kinetic properties indicate that the activation is caused by an SDS-dependent conformational change that facilitates the substrate accessibility to the dicopper active site.

FEMS Microbiol Lett, 2002 Feb 19, 208(1), 1 - 7
Diversity and phylogenetic analysis of bacteria in the mucosa of chicken ceca and comparison with bacteria in the cecal lumen; Gong J et al.; We reported the first attempt to describe mucosa-associated bacterial populations in the chicken ceca by molecular analysis of 16S rRNA genes . Bacteria in the mucosa were highly diverse but mainly Gram-positive with low G+C . Fusobacterium prausnitzii and butyrate-producing bacteria comprised the largest groups among 116 cloned sequences . Twenty five percent of the clones had less than 95% homology to database sequences . Many sequences were related to those of uncultured bacteria identified in human feces or the bovine rumen . Terminal restriction fragment length polymorphism (T-RFLP) analysis revealed some differences between bacterial populations present in the mucosa and lumen of ceca . Greater resolution of bacterial population was obtained using a culture-independent approach rather than a culture-based approach.

Int J Antimicrob Agents, 2002 Mar, 19(3), 233 - 6
Pharmacokinetics of teicoplanin in patients undergoing chronic haemodialysis; Papaioannou MG et al.; In order to define a dose regimen of teicoplanin for patients undergoing chronic haemodialysis so that they achieved trough drug serum levels above 10 mg/l, two single doses of 5 and 10 mg/kg were administered intravenously in seven anuric patients immediately after the end of haemodialysis . Concentrations of teicoplanin were determined by a microbiological assay in samples collected from peripheral veins via the arterial and the venous lines of the fistulae and from the dialysate during haemodialysis . The administration of a 5 and 10 mg/kg dose gave mean C(max) of 62.80 and 122.43 mg/l, mean AUC of 526.43 and 1103.98 mg h/l, mean half life (t(1/2)) of 109.09 and 107.06 h, mean clearance rates of 12.85 and 12.44 ml/min, mean apparent volumes of distribution of 1.68 and 1.68 l/kg and mean volumes of distribution at steady state of 0.31 and 0.28 l/kg, respectively . Trough serum levels above 10 mg/l were found for 24 h after the administration of the 5 mg/kg dose and for 48 h after the administration of the 10 mg/kg dose . Teicoplanin was not detected in the dialysate . Its concentrations in both the arterial and the venous lines of the fistulae were similar . Based on the time period after the administration of teicoplanin where the desired trough serum levels were found and on the observed t(1/2), it is proposed that teicoplanin should be administered at a dose of 10 mg/kg at 48-72 h intervals, in patients undergoing chronic haemodialysis for the therapy of infections caused by Gram-positive cocci.

Int J Syst Evol Microbiol, 2002 Mar, 52(Pt 2), 617 - 9
Arcanobacterium hippocoleae sp . nov., from the vagina of a horse; Hoyles L et al.; A polyphasic taxonomic study was performed on a previously unidentified gram-positive, facultatively anaerobic, diphtheroid-shaped organism isolated from a vaginal discharge of a horse . Comparative 16S rRNA gene sequencing demonstrated that the strain was a member of the genus Arcanobacterium, but sequence divergence values of >4% with described species of this genus (viz: Arcanobacterium haemolyticum, Arcanobacterium bernardiae, Arcanobacterium phocae, Arcanobacterium pluranimalium and Arcanobacterium pyogenes) demonstrated that the isolate represented a novel species . The unknown bacterium was readily distinguished from other Arcanobacterium species by biochemical tests . Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Arcanobacterium hippocoleae sp . nov . The type strain of A . hippocoleae is CCUG 44697T (= CIP 106850T).

Int J Syst Evol Microbiol, 2002 Mar, 52(Pt 2), 569 - 71
Arthrobacter nasiphocae sp . nov., from the common seal (Phoca vitulina); Collins MD et al.; An unknown gram-positive, catalase-positive, strictly aerobic, rod-shaped bacterium was isolated from the nasal cavities of two common seals . Chemical analysis revealed the presence in the bacterium of a hitherto unknown cell-wall murein {type: L-Lys-L-Ala2-Gly(2-3)-L-Ala (Gly)} . Comparative 16S rRNA gene sequencing showed that the unidentified rod was related to the Arthrobacter group of organisms, although sequence divergence values of >3% from established members of this genus indicated that it represents a novel species . On the basis of phenotypic and phylogenetic considerations, it is proposed that the unknown bacterium from seals (Phoca vitulina) be classified as a novel species, Arthrobacter nasiphocae sp . nov . The type strain of Arthrobacter nasiphocae is CCUG 42953T.

BMC Bioinformatics . 2002 Apr 03;3(1):10.
RNA:(guanine-N2) methyltransferases RsmC/RsmD and their homologs revisited--bioinformatic analysis and prediction of the active site based on the uncharacterized Mj0882 protein structure; Bujnicki JM et al.; BACKGROUND: Escherichia coli guanine-N2 (m2G) methyltransferases (MTases) RsmC and RsmD modify nucleosides G1207 and G966 of 16S rRNA . They possess a common MTase domain in the C-terminus and a variable region in the N-terminus . Their C-terminal domain is related to the YbiN family of hypothetical MTases, but nothing is known about the structure or function of the N-terminal domain . RESULTS: Using a combination of sequence database searches and fold recognition methods it has been demonstrated that the N-termini of RsmC and RsmD are related to each other and that they represent a "degenerated" version of the C-terminal MTase domain . Novel members of the YbiN family from Archaea and Eukaryota were also indentified . It is inferred that YbiN and both domains of RsmC and RsmD are closely related to a family of putative MTases from Gram-positive bacteria and Archaea, typified by the Mj0882 protein from M . jannaschii (1dus in PDB) . Based on the results of sequence analysis and structure prediction, the residues involved in cofactor binding, target recognition and catalysis were identified, and the mechanism of the guanine-N2 methyltransfer reaction was proposed . CONCLUSIONS: Using the known Mj0882 structure, a comprehensive analysis of sequence-structure-function relationships in the family of genuine and putative m2G MTases was performed . The results provide novel insight into the mechanism of m2G methylation and will serve as a platform for experimental analysis of numerous uncharacterized N-MTases.

J Am Chem Soc, 2002 Apr 10, 124(14), 3656 - 60
The first total synthesis of lipid II: the final monomeric intermediate in bacterial cell wall biosynthesis; VanNieuwenhze MS et al.; Bacterial peptidoglycan is composed of a network of beta-{1,4}-linked glyan strands that are cross-linked through pendant peptide chains . The final product, the murein sacculus, is a single, covalently closed macromolecule that precisely defines the size and shape of the bacterial cell . The recent increase in bacterial resistance to cell wall active agents has led to a resurgence of activity directed toward improving our understanding of the resistance mechanisms at the molecular level . The biosynthetic enzymes and their natural substrates can be invaluable tools in this endeavor . While modern experimental techniques have led to isolation and purification of the biosynthetic enzymes utilized in peptidoglycan biosynthesis, securing useful quantities of their requisite substrates from natural substrates has remained problematic . In an effort to address this issue, we report the first total synthesis of lipid II (4), the final monomeric intermediate utilized by Gram positive bacteria for peptidoglycan biosynthesis.

J Int Med Res, 2002, 30 Suppl 1, 20A - 30A
Experience with ampicillin/sulbactam in severe infections; Kanra G; The emergence of beta-lactamase-mediated resistance to established beta-lactam antibiotics prompted the development of beta-lactamase inhibitors for co-administration . Ampicillin has been combined with sulbactam for both parenteral and oral (as the mutual pro-drug sultamicillin) administration . The combination is active in vitro against a wide variety of Gram-positive and Gram-negative pathogens, including aerobic and anaerobic organisms . In clinical trials, ampicillin/sulbactam has proved clinically and bacteriologically effective against a variety of frequently encountered pediatric infections, including mild-to-moderate upper respiratory tract infections (acute otitis media, sinusitis, pharyngitis, and tonsillitis), severe post-operative and intra-abdominal infections, periorbital infections (which, left untreated, can lead to blindness, brain abscess, or death), acute epiglottitis, bacterial meningitis, and brain abscess . Ampicillin/sulbactam has also proved effective in the prevention of post-operative surgical infections in pediatric patients . The clinical efficacy profile of ampicillin/sulbactam and sultamicillin, combined with their excellent tolerability profile, make these agents attractive options for the management of many life-threatening infections in pediatric patients.

Nucleic Acids Res, 2002 Apr 1, 30(7), 1646 - 55
Sequence requirements for terminators and antiterminators in the T box transcription antitermination system: disparity between conservation and functional requirements; Grundy FJ et al.; The T box transcription termination control system is used in Gram-positive bacteria to regulate expression of aminoacyl-tRNA synthetase and other amino acid-related genes . Readthrough of a transcriptional terminator located in the leader region of the target gene is dependent on a specific interaction between the nascent leader transcript and the cognate uncharged tRNA . This interaction is required for formation of an antiterminator structure in the leader, which prevents formation of a competing transcriptional terminator stem-loop . The antiterminators and terminators of genes in this family are highly conserved in both secondary structure and primary sequence; the antiterminator contains the T box sequence, which is the most highly conserved leader element . These conserved features were investigated by phylogenetic and mutational analysis . Changes at highly conserved positions in the bulge and in the helix above the bulge reduced function, while alteration of other positions that were as much as 96% conserved did not have a major effect . The disparity between sequence conservation and function may be due to the requirement for maintaining base pairing in both the antiterminator and terminator structures.

J Bacteriol, 2002 Apr, 184(8), 2260 - 72
Evolutionary analysis by whole-genome comparisons; Bansal AK et al.; A total of 37 complete genome sequences of bacteria, archaea, and eukaryotes were compared . The percentage of orthologous genes of each species contained within any of the other 36 genomes was established . In addition, the mean identity of the orthologs was calculated . Several conclusions result: (i) a greater absolute number of orthologs of a given species is found in larger species than in smaller ones; (ii) a greater percentage of the orthologous genes of smaller genomes is contained in other species than is the case for larger genomes, which corresponds to a larger proportion of essential genes; (iii) before species can be specifically related to one another in terms of gene content, it is first necessary to correct for the size of the genome; (iv) eukaryotes have a significantly smaller percentage of bacterial orthologs after correction for genome size, which is consistent with their placement in a separate domain; (v) the archaebacteria are specifically related to one another but are not significantly different in gene content from the bacteria as a whole; (vi) determination of the mean identity of all orthologs (involving hundreds of gene comparisons per genome pair) reduces the impact of errors in misidentification of orthologs and to misalignments, and thus it is far more reliable than single gene comparisons; (vii) however, there is a maximum amount of change in protein sequences of 37% mean identity, which limits the use of percentage sequence identity to the lower taxa, a result which should also be true for single gene comparisons of both proteins and rRNA; (viii) most of the species that appear to be specifically related based upon gene content also appear to be specifically related based upon the mean identity of orthologs; (ix) the genes of a majority of species considered in this study have diverged too much to allow the construction of all-encompassing evolutionary trees . However, we have shown that eight species of gram-negative bacteria, six species of gram-positive bacteria, and eight species of archaebacteria are specifically related in terms of gene content, mean identity of orthologs, or both.

Biol Pharm Bull, 2002 Mar, 25(3), 367 - 71
In vitro and in vivo effects of macrophage-stimulatory polysaccharide from leaves of Perilla frutescens var . crispa; Kwon KH et al.; The crude polysaccharide (PFB-1) was isolated from the leaves of Perilla frutescens var . crispa by the sequential procedures with hot-water extraction, methanol reflux, and ethanol precipitation . It was further purified by anion column chromatography in order to obtain the partially purified polysaccharide (PFB-1-0) . In the presence of PFB-1-0, strong cellular lysosomal enzyme activity of murine peritoneal macrophages was observed in vitro . Compared to bacterial lipopolysaccharide (LPS), its activity was relatively high . The in vitro phagocytic activity was enhanced by PFB-1-0 as the similar pattern in both gram-negative bacteria, E . coli, and gram-positive bacteria, S . aureus with a time-dependent manner . We also investigated the production of several mediators by murine peritoneal macrophages upon stimulation with PFB-1 (in vivo) or PFB-1-0 (in vitro) . The levels of nitric oxide (NO) and tumor necrosis factor (TNF)-alpha were increased in the presence of PFB-1-0 in vitro . The PFB-1 stimulated the production of interleukin (IL)-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) in vivo . Results suggest that the polysaccharide from P . frutescens var . crispa represents an immunopotentiator and biological response modifiers in vitro and in vivo levels.

J Nat Prod, 2002 Mar, 65(3), 399 - 402
Phomadecalins A-D and phomapentenone A: new bioactive metabolites from Phoma sp . NRRL 25697, a fungal colonist of Hypoxylon stromata; Che Y et al.; Five new natural products, phomadecalins A-D (1-4) and phomapentenone A (5), have been obtained from cultures of Phoma sp . (NRRL 25697), a fungal colonist isolated from the stromata of Hypoxylon sp . The structures of these compounds were elucidated through a series of 1D and 2D NMR experiments . Compounds 1-4 display activity against Gram-positive bacteria.

Surg Neurol, 2002 Feb, 57(2), 81 - 6
Spontaneous disc space infections in adults; Friedman JA et al.; BACKGROUND: Spontaneous discitis typically affects children; much less is known about the disease in adults . We examined the clinical characteristics and the role of surgery in spontaneous infectious discitis in adults . METHODS: Twenty-nine consecutive adult patients (16 men, 13 women) with spontaneous infectious discitis were treated by a single surgeon (MJE) over a 5-year period . These patients were compared to 19 consecutive patients with postoperative discitis over the same time period . Mean follow-up was 2.4 years . RESULTS: The average age of patients with spontaneous discitis was 69.0 years . Eleven of these patients (38%) were diabetic and 9 (31%) had a known concurrent infection . Infections occurred at cervical (10%), thoracic (34%), and lumbar interspaces (59%), and at multiple disc spaces in eight cases (28%) . Ten patients (34%) had an elevated serum leukocyte count and 21 patients (72%) had an elevated erythrocyte sedimentation rate . While Gram-positive infection was most common, a broad spectrum of microbes was isolated . All patients were treated with i.v . antibiotics for a mean duration of 6 weeks . Four patients required surgical fusion; the rest were treated with external immobilization . At follow-up, 21 patients (72%) were symptom-free, 3 patients (10%) had ongoing disease, and 5 patients (17%) had died of unrelated causes . Compared to patients with postoperative discitis treated over the same time period, spontaneous discitis in adults affected older patients and required a broader spectrum of antibiotic coverage . Outcomes were similar between the two groups . CONCLUSIONS: While spontaneous discitis has predominantly been reported in children, the disease also affects older adults . Spontaneous discitis in adults is associated with advanced age, diabetes mellitus, and systemic infection . Elevated serum leukocyte count lacks diagnostic sensitivity . A high cure rate is achieved with antibiotics and external immobilization . Surgical treatment is rarely required.

Folia Microbiol (Praha), 2001, 46(6), 483 - 7
An origin of DNA replication from streptomycete phage phi U1; Farkasovska J et al.; A DNA fragment from phage phi U1 containing an origin of DNA replication was identified . This fragment, designated ori, was able to support the maintenance in Streptomyces lividans of a plasmid lacking a functional Gram-positive ori . The sequence of the minimal ori fragment was determined and analyzed . The minimal fragment conferring replication origin function contained a number of direct and inverted repeats . The absence of an open reading frame in this ori fragment indicates that host factors alone were sufficient to initiate replication at ori.

J Am Chem Soc, 2002 Mar 20, 124(11), 2436 - 7
Corynebactin and enterobactin: related siderophores of opposite chirality; Bluhm ME et al.; Most species of bacteria employ siderophores to acquire iron . 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 isolated from Gram-negative bacteria, but contains L-theronine 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 synthesized . The chirality and relative conformational stability of the three ferric complexes of enterobactin, corynebactin, and the hybrid has been investigated . In contrast to enterobactin, corynebactin assumes a Lambda configuration . However, the ferric serine-corynebactin hybrid forms a racemic mixture, only slightly favoring the Lambda conformation.

Am J Crit Care, 2002 Mar, 11(2), 141 - 9
Bacterial growth in secretions and on suctioning equipment of orally intubated patients: a pilot study; Sole ML et al.; BACKGROUND: Contamination of equipment, colonization of the oropharynx, and microaspiration of secretions are causative factors for ventilator-associated pneumonia . Suctioning and airway management practices may influence the development of ventilator-associated pneumonia . OBJECTIVES: To identify pathogens associated with ventilator-associated pneumonia in oral and endotracheal aspirates and to evaluate bacterial growth on oral and endotracheal suctioning equipment . METHODS: Specimens were collected from 20 subjects who were orally intubated for at least 24 hours and required mechanical ventilation . At baseline, oral and sputum specimens were obtained for culturing, and suctioning equipment was changed . Specimens from the mouth, sputum, and equipment for culturing were obtained at 24 hours (n=18) and 48 hours (n=10) . RESULTS: After 24 hours, all subjects had potential pathogens in the mouth, and 67% had sputum cultures positive for pathogens . Suctioning devices were colonized with many of the same pathogens that were present in the mouth . Nearly all (94%) of tonsil suction devices were colonized within 24 hours . Most potential pathogens were gram-positive bacteria . Gram-negative bacteria and antibiotic-resistant organisms were also present in several samples . CANCLUSIONS: The presence of pathogens in oral and sputum specimens in most patients supports the notion that microaspiration of secretions occurs . Colonization is a risk factor for ventilator-associated pneumonia . The equipment used for oral and endotracheal suctioning becomes colonized with potential pathogens within 24 hours . It is not known if reusable oral suction equipment contributes to colonization; however, because many bacteria are exogenous to patients' normal flora, equipment may be a source of cross-contamination.

Compend Contin Educ Dent, 2001 Dec, 22(12), 1093 - 6, 1098, 1100-2 passim; quiz 1108
Comparison of the effectiveness of several denture sanitizing systems: a clinical study; Glass RT et al.; The purpose of this clinical study was to test the effectiveness of three methods of decontamination on complete dentures . Dentures worn by patients for varying lengths of time were handled aseptically and treated with three different treatment modalities . The dentures were touched and sectioned and then retouched to a variety of microbiological media . The quantity of microbial growth was recorded and predominating microorganisms were identified using standard microbiological techniques . System A was found to consistently decontaminate and sanitize dentures worn by patients . System B and System C showed variable reduction of microorganisms . An unexpected spectrum of both pathogenic and opportunistic microorganisms was found in the dentures examined, including a wide range of gram-negative bacteria, gram-positive bacteria, and yeasts . A wide range of microorganisms must be considered when treating either oral or systemic diseases in denture wearers . Denture hygiene and decontamination are critical to the prevention of oral and systemic disease transmission . The dentures of ill patients must be considered as possible sources of pathogenic microorganisms.

Acta Gastroenterol Belg, 2001 Oct-Dec, 64(4), 318 - 20
Ileo-caecal actinomycosis: report of a case simulating complicated inflammatory bowel disease; Postal A et al.; Abdominal actinomycosis is a rare infectious disease caused by Actinomyces israelii, a gram-positive anaerobic saprophyte germ that is a normal inhabitant of the upper intestinal tract in humans . Actinomyces israelii rarely cause abdominal infections or actinomycosis . Abdominal actinomysosis is characterised by fistulae and abscesses and may mimic cancer or inflammatory bowel disease . Abdominal actinomycosis is difficult to diagnose preoperatively, and often require surgical removal of the diseased tissue, allowing pathologists for giving the definitive diagnosis, revealed by characteristic "sulfur granules" . The authors report herein the case of a 47-year-old man who presented with diarrhoea and abdominal pain . Abdominal computed tomography evoked complicated inflammatory bowel disease and surgical procedure was decided . Laparoscopic exploration did not provide further significant information, and laparotomy with diseased bowel resection was performed . Pathology demonstrated "sulfur granules" and allowed the diagnosis of abdominal actinomycosis . This case demonstrated that abdominal actinomycosis should be included in the differential diagnosis when computed tomography shows an infiltrative and inflammatory mass.

Res Microbiol, 2002 Jan-Feb, 153(1), 45 - 52
Bacteriocin production by Actinobacillus actinomycetemcomitans isolated from the oral cavity of humans with periodontal disease, periodontally healthy subjects and marmosets; Lucia LF et al.; The ability of Actinobacillus actinomycetemcomitans to produce bacteriocin has rarely been reported . Antagonistic substance production may confer an important ecological advantage for the producer microorganisms, especially in a competitive ecosystem such as the oral cavity . In the present study, 75 A . actinomycetemcomitans strains isolated from the oral cavity of human patients with periodontal disease, periodontally healthy subjects and marmosets, as well as two reference strains (A . actinomycetemcomitans ATCC 29523 and FDC Y4) were evaluated for auto-, iso-, and heteroantagonistic activity . Fifty-one (68.00%) strains exhibited antagonistic activity; heteroantagonism was observed more often than isoantagonism . Isolated strains antagonized 17 different species of gram-positive and gram-negative bacteria from the oral and nonoral microbiota . Sensitivity to heat and to proteolytic enzymes constituted strong evidence that the antagonistic substance has a proteic nature . Taken together, our data enabled us to confirm that the antagonistic substance detected was a bacteriocin . The wide spectrum of activity indicates the possibility that more than one antagonistic substance is produced and that these substances play an important role in the ecological balance of the oral ecosystem.

J Biol Chem, 2002 May 17, 277(20), 17448 - 56 Epub 2002 Mar 04.
Toll-like receptor 2 and 4 (TLR2 and TLR4) agonists differentially regulate secretory interleukin-1 receptor antagonist gene expression in macrophages; Carl VS et al.; Treatment of macrophages with lipopolysaccharide (LPS) from Gram-negative bacteria or peptidoglycan (PGN) from Gram-positive bacteria activates multiple intracellular signaling pathways and a large, diverse group of nuclear transcription factors . The signaling receptors for PGN and LPS are now known to be the Toll-like receptors 2 and 4 (TLR2 and -4, respectively) . While a large body of literature indicates that the members of the TLR family activate nearly identical cytoplasmic signaling programs, several recent reports have suggested that the functional outcomes of signaling via TLR2 or TLR4 are not equivalent . In the current studies, we compared the responses of the secretory IL-1 receptor antagonist (sIL-1Ra) gene to both LPS and PGN . Both LPS and PGN induced IL-1Ra gene expression; however, the combination of both stimuli synergistically increased sIL-1Ra mRNA expression and promoter activity, suggesting that the signals induced by PGN and LPS are not equivalent . While both LPS and PGN utilized the PU.1-binding sites in the proximal sIL-1Ra promoter region to generate a full response, additional distinct promoter elements were utilized by LPS or PGN . Activation of p38 stress-activated protein kinase was required for LPS- or PGN-induced IL-1Ra gene expression, but the p38-responsive promoter elements localized to distinct regions of the sIL-1Ra gene . Additionally, while the LPS-induced, p38-dependent response was dependent upon PU.1 binding, the PGN-induced, p38 response was not . Collectively, these data indicated that while some of the intracellular signaling events by TLR2 and TLR4 agonists are similar, there are clearly distinct differences in the responses elicited by these two bacterial products.

Syst Appl Microbiol, 2001 Dec, 24(4), 623 - 33
Microbial diversity of cultivatable bacteria associated with the North Sea bryozoan Flustra foliacea; Pukall R et al.; The microbial diversity of cultivatable bacteria associated with the bryozoan species Flustra foliacea from the North Sea was investigated by a molecular approach . Amplified ribosomal RNA restriction analyses (ARDRA) and 16S rDNA partial sequence analysis revealed differences in the composition of cultivatable bacteria populations from single bryozoan colonies collected from two different sampling sites in the North Sea as well from one site taken at different points in time . Whereas gamma-Proteobacteria identified as Shewanella frigidimarina, Pseudoalteromonas ssp . and Psycbrobacter ssp . were predominant on samples of Flustra I (taken near the island of Helgoland), most bacteria isolated from Flustra II, originating from the Steingrund, could be affiliated to Gram-positive taxa . Survey of the bryozoan samples from the latter site in February 2000 led to the detection of a phylogenetically mixed bacterial population, consisting of gamma-, and alpha-Proteobacteria and Gram-positive bacteria with low and high GC-content (Flustra III) . As these bacteria are among the most widely isolated organisms from the marine environment, it may be concluded that the bryozoan Flustra foliacea accepts colonization of surfaces by bacteria which are common inhabitants of the marine environment and which may have been transferred into this environment from terrestrial sites.

Sci Total Environ, 2002 Feb 21, 285(1-3), 3 - 10
Influence of nonylphenol on the microbial community of lake sediments in microcosms; Jontofsohn M et al.; In this study the impact of nonylphenol, an estrogenic degradation product of alkylphenol polyethoxylates, on the microbial community structure in contaminated sediments of aquatic microcosms using in situ hybridization with fluorescently labeled oligonucleotides probes was investigated . A positive correlation between nonylphenol concentration and cell numbers of bacteria and microfungi as well as an increase in the numbers of active bacteria was found . However, the ratio between total microorganisms and active bacteria remained unchanged . A large fraction of the cells could be identified using group specific oligonucleotide probes . A slight change in the composition of the microbial community structure was observed, with Gram-positive bacteria with high DNA G + C-content becoming more abundant at higher concentrations of nonylphenol.

Appl Environ Microbiol, 2002 Mar, 68(3), 1082 - 7
Simultaneous extraction from bacterioplankton of total RNA and DNA suitable for quantitative structure and function analyses; Weinbauer MG et al.; The aim of this study was to develop a protocol for the simultaneous extraction from bacterioplankton of RNA and DNA suitable for quantitative molecular analysis . By using a combined mechanical and chemical extraction method, the highest RNA and DNA yield was obtained with sodium lauryl sarcosinate-phenol or DivoLab-phenol as the extraction mix . The efficiency of extraction of nucleic acids was comparatively high and varied only moderately in gram-negative bacterial isolates and bacterioplankton (RNA, 52 to 66%; DNA, 43 to 61%); significant amounts of nucleic acids were also obtained for a gram-positive bacterial isolate (RNA, 20 to 30%; DNA, 20 to 25%) . Reverse transcription-PCR and PCR amplification products of fragments of 16S rRNA and its genes were obtained from all isolates and communities, indicating that the extracted nucleic acids were intact and pure enough for community structure analyses . By using single-strand conformation polymorphism of fragments of 16S rRNA and its gene, community fingerprints were obtained from pond bacterioplankton . mRNA transcripts encoding fragments of the enzyme nitrite reductase gene (nir gene) could be detected in a pond water sample, indicating that the extraction method is also suitable for studying gene expression . The extraction method presented yields nucleic acids that can be used to perform structural and functional studies of bacterioplankton communities from a single sample.

Hokkaido Igaku Zasshi, 2002 Jan, 77(1), 3 - 7
{Recent advances in innate immune host defense--hose defense lectins and endotoxin receptors}; Kuroki Y; Accumulating evidence has revealed that CD14 and Toll-like receptors (TLRs) function as pattern recognition receptors for broad ranges of pathogens . The structural characteristics of these proteins is that they possess leucine-rich repeats, which appear to be involved in protein-protein interaction . We have shown that the extracellular TLR2 domain directly binds to peptidoglycan (PGN) which is abundant in the cell wall of Gram-positive bacteria and that the N-terminal region of TLR2 is critical for the recognition of PGN . Surfactant proteins A and D (pulmonary collectins) function as host defense lectins and play important roles in innate immunity in the lung-Collectins including SP-A, SP-D and mannose-binding proteins can bind CD14 . We have demonstrated that SP-A modulates the cellular response to smooth and rough lipopolysaccharides (LPS) by interaction with CD14 . In addition, this protein has been revealed to function as a negative regulator against PGN-induced TNF-alpha secretion from macrophages through direct interaction with TLR2 . We propose that SP-A modulates inflammatory responses against the bacterial components by interaction with pattern-recognition receptors.

J Leukoc Biol, 2002 Mar, 71(3), 503 - 10
Toll-like receptor 2 (TLR2) mediates activation of stress-activated MAP kinase p38; Vasselon T et al.; Early events in the response of cells to lipopolysaccharide (LPS) include activation of NF-kappaB and stress-activated MAP kinase p38 . Recent studies have shown that the human Toll-like receptor 2 (TLR2) mediates activation of NF-kappaB in response to commercial preparations of LPS (comLPS), membrane lipoproteins, and Gram-positive bacterial products . Here, we show that expression of TLR2 in human embryonic kidney 293 cells enabled p38 phosphorylation in response to comLPS, a synthetic bacterial lipoprotein, and B . subtilis . Activation of p38 was confirmed by an in vitro kinase assay using ATF2 as substrate and by an assay measuring activation of the downstream effector of p38, MAP kinase-activated protein kinase in cells . Thus, TLR2 initiated the signaling pathway for p38 in response to bacterial products.

World J Surg, 2002 Mar, 26(3), 372 - 6 Epub 2001 Nov 15.
Characteristics of infection with Candida species in patients with necrotizing pancreatitis; Isenmann R et al.; This study focuses on the relevance of Candida infection (albicans and non-albicans) in patients with necrotizing pancreatitis . Altogether, 92 patients with infected pancreatic necrosis were reviewed for Candida infection . All patients underwent surgical necrosectomy for infected pancreatic necrosis . Data from patients with Candida growth in intraoperative smears were compared to those obtained from patients without Candida infection . There were 22 patients (24%) with Candida infection . Patients with or without Candida infection were comparable regarding age, gender, etiology, and severity scores at admission . Candida patients suffered a higher mortality (64% vs.19%, p = 0.0001) and experienced more systemic complications (3.2 +/- 1.6 vs . 2.1 +/- 1.4; p= 0.004) than patients without Candida . Preoperative antibiotics were given significantly longer prior to Candida infection (19.0 +/- 13.2 vs . 6.4 +/- 10.3 days; p < 0.0001) . With regard to the concomitant spectrum of bacteria, solitary gram-negative infection was rare in Candida patients (5% vs . 43%, p =0.0006) . The presence of Candida in patients with infected pancreatic necrosis is associated with increased mortality . Our data provide evidence that application of antibiotics contributes to the development of Candida infection and to changes in the bacterial spectrum of infected necrosis with an increase in the incidence of gram-positive infection.

Clin Microbiol Infect, 1997 Feb, 3(1), 24 - 31
Is TNF-alpha a prognostic factor in patients with sepsis?
Schaumann R, Schlick T, Schaper M, Shah PM.
OBJECTIVE: To determine tumor necrosis factor-alpha (TNF-alpha) levels in a prospective study in 58 hospitalized patients in a department of internal medicine (63 episodes, 29 in immunocompromised patients) during a 7-month period . METHODS: Patients fulfilling the following criteria were included: clinical evidence of acute infection, temperature >38.2 degrees C, tachycardia >90 beats/min, tachypnea >20 breaths/min . Samples were taken from day 1 up to day 13 after an infection was diagnosed, and TNF-alpha was determined by enzyme immunoassay . RESULTS: In 29 episodes (46.0%) the infection was microbiologically documented . The median of the TNF-alpha levels in the Gram-negative episodes was significantly higher than that in the Gram-positive episodes (p=0.002) . Thirteen of 63 episodes (20.6%) had a fatal outcome . With respect to all measured values, the non-survivors had a significantly higher median of TNF-alpha levels than the survivors (p=0.0001) . There was, however, great interpatient and intrapatient variability in TNF-alpha levels; thus, no unequivocal correlation between TNF-alpha and outcome could be documented . CONCLUSIONS: Our data indicate that the influence of the infecting organism on TNF-alpha kinetics is less pronounced than that of the underlying disease.

J Antibiot (Tokyo), 2001 Dec, 54(12), 1036 - 44
Goadsporin, a chemical substance which promotes secondary metabolism and morphogenesis in streptomycetes . I . Purification and characterization; Onaka H et al.; Streptomycetes, which belong to the Gram-positive bacteria, produce secondary metabolites and sporulate . The timing of starting the secondary metabolite production and the sporulation depends on environmental conditions such as nitrogen and carbon sources . In order to obtain a tool for understanding the regulation mechanism, we carried out screening for chemical substances that induce secondary metabolism and sporulation in streptomycetes and found an active substance from the culture broth of Streptomyces sp . TP-A0584 . This substance designated goadsporin promoted the formation of red pigment and sporulation at a concentration of 1 microM in Streptomyces lividans TK23 which does not produce the pigment under normal growth conditions . Goadsporin is an oligopeptide consisting of 19 amino acids with the molecular formula C72H97N19020S2 . Sporulation and/or secondary metabolite production was induced in 36 streptomycetes strains among 42 strains tested . These results suggest that goadsporin acts on a common regulation pathway for sporulation and secondary metabolism in streptomycetes and can be a powerful tool to analyze the regulation mechanism.

Am Fam Physician, 2002 Feb 1, 65(3), 455 - 64
Quinolones: a comprehensive review; Oliphant CM et al.; With the recent introduction of agents such as gatifloxacin and moxifloxacin, the traditional gram-negative coverage of fluoroquinolones has been expanded to include specific gram-positive organisms . Clinical applications beyond genitourinary tract infections include upper and lower respiratory infections, gastrointestinal infections, gynecologic infections, sexually transmitted diseases, and some skin and soft tissue infections . Most quinolones have excellent oral bioavailability, with serum drug concentrations equivalent to intravenous administration . Quinolones have few adverse effects, most notably nausea, headache, dizziness, and confusion . Less common but more serious adverse events include prolongation of the corrected QT interval, phototoxicity, liver enzyme abnormalities, arthropathy, and cartilage and tendon abnormalities . The new fluoroquinolones are rarely first-line agents and should be employed judiciously . Inappropriate use of agents from this important class of antibiotics will likely worsen current problems with antibiotic resistance . Applications of fluoroquinolones in biologic warfare are also discussed.

Acta Crystallogr D Biol Crystallogr, 2002 Mar, 58(Pt 3), 515 - 8 Epub 2002 Feb 21.
Crystallization, preliminary X-ray analysis and biophysical characterization of HPr kinase/phosphatase of Mycoplasma pneumoniae; Steinhauer K et al.; The Mycoplasma pneumoniae HPr kinase/phosphatase (HPrK/P) is a member of a large family of enzymes which are central to carbon regulation in Gram-positive bacteria . The full-length M . pneumonia HPrK/P was crystallized from solutions of polyethylene glycol 8000 and KCl or NaCl which also contained the non-hydrolysable ATP analog adenosine 5'-{beta, gamma-methylene}triphosphate (AMPPCP) . The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 117.1, b = 127.7, c = 170.7 A . A complete X-ray intensity data set has been collected and processed to 2.50 A resolution . The slow self-rotation function revealed the presence of a sixfold axis . Dynamic light-scattering (DLS) experiments indicated a molecular weight of 197 kDa for HPrK/P in the absence of AMPPCP and of 217 kDa in the presence of the ATP analog . Thus, the biophysical and crystallographic data suggest that HPrK/P is a functional hexamer that undergoes an ATP-binding-induced conformational change.

J Surg Res, 2002 Mar, 103(1), 89 - 95
Surgical site infections: reanalysis of risk factors; Malone DL et al.; BACKGROUND: Surgical site infections (SSI) are the most common nosocomial infection in surgical patients, accounting for 38% of all such infections, and are a significant source of postoperative morbidity resulting in increased hospital length of stay and increased cost . During 1986-1996 the Center for Disease Control and Prevention's National Nosocomial Infections Surveillance system reported 15,523 SSI following 593,344 operations (2.6%) . Previous studies have documented patient characteristics associated with an increased risk of SSI, including diabetes, tobacco or steroid use, obesity, malnutrition, and perioperative blood transfusion . In this study we sought to reevaluate risk factors for SSI in a large cohort of noncardiac surgical patients . METHODS: Prospective data (NSQIP) were collected on 5031 noncardiac surgical patients at the Veteran's Administration Maryland Healthcare System from 1995 to 2000 . All preoperative risk factors were evaluated as independent predictors of surgical site infection . RESULTS: The mean age of the study cohort was 61 plus minus 13 . SSI occurred in 162 patients, comprising 3.2% of the study cohort . Gram-positive organisms were the most common bacterial etiology . Multiple logistic regression analysis documented that diabetes (insulin- and non-insulin-dependent), low postoperative hematocrit, weight loss (within 6 months), and ascites were significantly associated with increased SSI . Tobacco use, steroid use, and chronic obstructive pulmonary disease (COPD) were not predictors for SSI . CONCLUSION: This study confirms that diabetes and malnutrition (defined as significant weight loss 6 months prior to surgery) are significant preoperative risk factors for SSI . Postoperative anemia is a significant risk factor for SSI . In contrast to prior analyses, this study has documented that tobacco use, steroid use, and COPD are not independent predictors of SSI . Future SSI studies should target early preoperative intervention and optimization of patients with diabetes and malnutrition.

Biochim Biophys Acta, 2002 Jan 15, 1569(1-3), 167 - 73
Single chain antibodies specific for fatty acids derived from a semi-synthetic phage display library; Gargir A et al.; The biological activities of many acylated molecules are lipid dependent . Lipids, however, are poorly immunogenic or non-immunogenic . We employed a phage display semi-synthetic human antibody library to isolate anti-lipid antibodies . Selection was done against methyl palmitate, a 16 carbon aliphatic chain, and a major component of bacterial glycolipids and lipoproteins in animal cells . The selected single chain variable fragment (scFv) bound specifically to a 16 carbon aliphatic chain and to a lesser extent to a 14 or 18 carbon aliphatic chain and poorly to either 12, 22 or 8 carbon aliphatic chains . Furthermore, the scFv prevented micelle formation of lipoteichoic acid from Gram-positive bacteria; inhibited lipopolysaccharide-induced tumor necrosis factor alpha release in mononuclear cells; bound to hydrophobic bacterial surfaces, especially those of Gram-positive bacteria, and bound to Lck, a mammalian palmitated lipoprotein . Our data suggest that the phage antibody library can be successfully employed to obtain human anti-aliphatic scFv human antibody fragment with potential therapeutic applications in neutralizing the deleterious effects of bacterial toxins as well as in structure--function analysis of lipoproteins in animal cells.

J Korean Med Sci, 2002 Feb, 17(1), 113 - 5
Penicillin-resistant Aerococcus viridans bacteremia associated with granulocytopenia; Uh Y et al.; Aerococcus viridans, a catalase-negative gram-positive coccus rarely causing bacteremia, was isolated from blood cultures of a 52-yr-old man under the granulocytopenic condition . The isolate showed the typical characteristics of A . viridans, i.e., tetrad arrangements in gram stain, positive pyrrolidonyl aminopeptidase (PYR) and negative leucine aminopeptidase (LAP) reactions, and no growth at 45 degrees C.The isolate was revealed to be highly resistant to penicillin, erythromycin, clindamycin, and ceftriaxone, although most strains of A . viridans isolated from the previously reported patients were susceptible to penicillin and other commonly used antibiotics . Even though A . viridans is rarely associated with human infections, it could be a potential causative agent of bacteremia, especially in immunocompromised patients.

J Clin Periodontol, 2002 Jan, 29(1), 62 - 72
Effect of the enamel matrix derivative Emdogain on the growth of periodontal pathogens in vitro; Spahr A et al.; OBJECTIVES: The aim of this study was to evaluate the effect of Emdogain (EMD), used for periodontal regeneration, on the growth of periodontal pathogens like Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia . For comparison, we studied the effect of EMD on several microbes associated with other oral diseases as well as its effect on non-pathogenic oral inhabitants . METHODS: Freshly prepared EMD or its vehicle propylene glycol alginate (PGA) alone were added to calibrated suspensions of microbes . As a control, imitating the post-surgical subgingival situation after flap closure, a serum/NaCl-solution mixture was used . Aliquots for growth assays were taken at scheduled times for calculation of colony-forming units and cell densities over an observation period of 24 h . Additionally, EMD was spotted onto selected, newly seeded microbes growing on agar plates to see if growth inhibition zones could be produced . RESULTS: The study revealed a marked inhibitory effect of EMD on the growth of the gram-negative periodontal pathogens . A . actinomycetemcomitans showed a significant decrease (p=0.012) in viable counts after 24 h when EMD was added at baseline . P . gingivalis and P . intermedia also showed a marked growth reduction in the presence of EMD and in these cultures no viable microbes could be detected anymore after 24 h . In contrast, no significant growth inhibition was observed in gram-positive bacteria . CONCLUSIONS: The results suggest that EMD has a positive effect on the composition of bacterial species in the post-surgical periodontal wound, by selectively restricting growth of periopathogens that could hamper the wound healing and reduce the outcome of regenerative procedures.

Bioorg Med Chem Lett, 2002 Feb 25, 12(4), 697 - 9
Benzylidene rhodanines as novel inhibitors of UDP-N-acetylmuramate/L-alanine ligase; Sim MM et al.; Benzylidene rhodanines are novel inhibitors of UDP N-acetylmuramate/L-alanine ligase . They showed selective whole-cell activity against the Gram-positive MRSA but not against the Gram-negative Escherichia coli . Their cytotoxic effect on mammalian CHO cells was also evaluated.

J Biol Chem, 2002 May 3, 277(18), 15271 - 6 Epub 2002 Feb 11.
Adenylyl cyclase Rv1264 from Mycobacterium tuberculosis has an autoinhibitory N-terminal domain; Linder JU et al.; Mycobacterium tuberculosis contains 15 class III adenylyl cyclase genes . The gene Rv1264 is predicted to be composed of two distinct protein modules . The C terminus seems to code for a catalytic domain belonging to a subfamily of adenylyl cyclase isozymes mostly found in Gram-positive bacteria . The expressed protein was shown to function as a homodimeric adenylyl cyclase (1 micromol of cAMP x mg(-1) x min(-1)) . In analogy to the structure of the mammalian adenylyl cyclase catalyst, six amino acids were targeted by point mutations and found to be essential for catalysis . The N-terminal region represents a novel protein domain, the occurrence of which is restricted to several adenylyl cyclases present in Gram-positive bacteria . The purified full-length enzyme was 300-fold less active than the catalytic domain alone . Thus, the N-terminal domain appeared to be autoinhibitory . The N-terminal domain contains three prominent polar amino acid residues (Asp(107), Arg(132), and Arg(191)) that are invariant in all seven sequences of this domain currently available . Mutation of Asp(107) to Ala relaxed the inhibition and resulted in a 6-fold increase in activity of the Rv1264 holoenzyme, thus supporting the role of this domain as a potential novel regulator of adenylyl cyclase activity.

Int J Syst Evol Microbiol, 2002 Jan, 52(Pt 1), 85 - 90
Dietzia psychralcaliphila sp . nov., a novel, facultatively psychrophilic alkaliphile that grows on hydrocarbons; Yumoto I et al.; A novel, facultatively psychrophilic alkaliphile that grows on a chemically defined medium containing n-alkanes as the sole carbon source was isolated from a drain of a fish product-processing plant . The isolate was an aerobic, non-motile, gram-positive bacterium . The bacterium was catalase-positive and oxidase-negative . The cell wall contained meso-diaminopimelic acid, arabinose and galactose; the glycan moiety of the cell wall contained acetyl residues . The G+C content of the DNA was 69.6 mol % . Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolate was closely related to members of the genus Dietzia (96.1-96.8% similarity) . Comparisons of phenotypic and chemotaxonomic characteristics between the isolate and the two known Dietzia species showed that they were very similar . However, the isolate differed from the two known Dietzia species in growth temperature range and certain physiological characteristics . DNA-DNA hybridization revealed that the isolate had 38.4 and 49.7% relatedness, respectively, to Dietzia maris and Dietzia natronolimnaea . On the basis of the physiological and biochemical characteristics, the phylogenetic position as determined by 16S rRNA gene analysis and DNA-DNA relatedness, it is concluded that the isolate should be designated as a novel species, for which the name Dietzia psychralcaliphila sp . nov . is proposed . The type strain is ILA-1T (= JCM 10987T = IAM14896T = NCIMB 13777T).

Int J Syst Evol Microbiol, 2002 Jan, 52(Pt 1), 77 - 84
Knoellia sinensis gen . nov., sp . nov . and Knoellia subterranea sp . nov., two novel actinobacteria isolated from a cave; Groth I et al.; Two novel strains of the class Actinobacteria were isolated from a cave in China . Cells of both strains were gram-positive, non-motile, non-spore-forming and not acid-fast and exhibited a rod/coccus growth cycle . Both isolates grew well on complex organic media under aerobic conditions . Their cell wall peptidoglycan contained meso-diaminopimelic acid as diagnostic diamino acid . The acyl type of the glycan chain of peptidoglycan was acetyl . The major respiratory quinone was MK-8(H4) . The cellular fatty acid profile was characterized by the predominance of 13-methyltetradecanoic (i-C15:0), 15-methylhexadecanoic (i-C17:0), 14-methylpentadecanoic (i-C16:0) and 14-methylhexadecanoic (ai-C17:0) acids . The major polar lipids were phosphatidylethanolamine, phosphatidylinositol and diphosphatidylglycerol . Mycolic acids were absent . The DNA G+C composition was 68-69 mol% . 16S rDNA-based phylogenetic analysis revealed an intermediate phylogenetic position of the cave isolates between the genera Janibacter and Tetrasphaera, which did not permit their unambiguous affiliation to either genus . Differences in morphological, physiological and chemotaxonomic properties between the two isolates and their closest phylogenetic neighbours support the proposal of a new genus and two novel species, Knoellia sinensis gen . nov., sp . nov . and Knoellia subterranea sp . nov . The type and only strains of the species are respectively HKI 0119T (= DSM 12331T = CIP 106775T) and HKI 0120T (= DSM 12332T = CIP 106776T).

Int J Syst Evol Microbiol, 2002 Jan, 52(Pt 1), 207 - 10
Isobaculum melis gen . nov., sp . nov., a Carnobacterium-like organism isolated from the intestine of a badger; Collins MD et al.; Phenotypic and phylogenetic studies were performed on a hitherto undescribed facultatively anaerobic, catalase-negative, gram-positive rod-shaped organism, strain M577-94T, isolated from the small intestine of a dead badger . It resembled carnobacteria in terms of its long-chain cellular fatty acid composition, but differed markedly from the latter in possessing a cell-wall murein based on L-lysine (type L-Lys-L-Thr-Gly) . Comparative 16S rRNA gene sequencing showed that the unknown bacterium represents a new line closely related to, albeit distinct from, the genera Carnobacterium and Desemzia . On the basis of phylogenetic and phenotypic evidence, it is proposed that strain M577-94T be classified as Isobaculum melis gen . nov., sp . nov . The type strain of Isobaculum melis is CCUG 37660T (= DSM 13760T).

Int J Syst Evol Microbiol, 2002 Jan, 52(Pt 1), 109 - 13
Alicyclobacillus herbarius sp . nov., a novel bacterium containing omega-cycloheptane fatty acids, isolated from herbal tea; Goto K et al.; A thermo-acidophilic gram-positive bacterium, strain CP-1T, which grows aerobically at 35-65 degrees C (optimum 55-60 degrees C) and at pH 3.5-6.0 (optimum pH 4.5-5.0), was isolated from a herbal tea made from the dried flowers of hibiscus . Phylogenetic analysis of the 16S rRNA gene sequences showed that this bacterium was clearly distinguishable from previously described species of the genera Alicyclobacillus and Sulfobacillus . Strain CP-1T had unique omega-cycloheptane fatty acids as the major membrane lipid component, a characteristic which is peculiar to Alicyclobacillus cycloheptanicus . However, phenotypic and chemotaxonomic characteristics of strain CP-1T were different from those of the type strain of A . cycloheptanicus . DNA-DNA hybridization between the type strains of Alicyclobacillus species and Sulfobacillus disulfidooxidans was <20%, indicating that strain CP-1T represents a distinct species . On the basis of these results, the name Alicyclobacillus herbarius is proposed for this organism . The type strain is strain CP-1T (= DSM 13609T = IAM 14883T = NRIC 0477T).

J Biol Chem, 2002 Apr 26, 277(17), 15028 - 34 Epub 2002 Feb 08.
Novel signal transduction pathway utilized by extracellular HSP70: role of toll-like receptor (TLR) 2 and TLR4; Asea A et al.; Recent studies have initiated a paradigm shift in the understanding of the function of heat shock proteins (HSP) . It is now clear that HSP can and do exit mammalian cells, interact with cells of the immune system, and exert immunoregulatory effects . We recently demonstrated that exogenously added HSP70 possesses potent cytokine activity, with the ability to bind with high affinity to the plasma membrane, elicit a rapid intracellular Ca(2+) flux, activate NF-kappaB, and up-regulate the expression of pro-inflammatory cytokines in human monocytes . Here for the first time, we report that HSP70-induced proinflammatory cytokine production is mediated via the MyD88/IRAK/NF-kappaB signal transduction pathway and that HSP70 utilizes both TLR2 (receptor for Gram-positive bacteria) and TLR4 (receptor for Gram-negative bacteria) to transduce its proinflammatory signal in a CD14-dependent fashion . These studies now pave the way for the development of highly effective pharmacological or molecular tools that will either up-regulate or suppress HSP70-induced functions in conditions where HSP70 effects are desirable (cancer) or disorders where HSP70 effects are undesirable (arthritis and arteriosclerosis).

Syst Appl Microbiol, 2001 Nov, 24(3), 362 - 7
Methanosarcina lacustris sp . nov., a new psychrotolerant methanogenic archaeon from anoxic lake sediments; Simankova MV et al.; A new psychrotolerant methanogenic archaeon strain ZS was isolated from anoxic lake sediments (Switzerland) . The cells of the organism were non-motile cocci, 1.5-3.5 microm in diameter . The cells aggregated and formed pseudoparenchyma . The cell wall was Gram-positive . The organism utilized methanol, mono-, di-, trimethylamine and H2/CO2 with methane production . The temperature range for growth was 1-35 degrees C with an optimum at 25 degrees C . The DNA G+C content of the organism was 43.4 . mol% . Analysis of the 16S rRNA gene sequence showed that strain ZS was phylogenetically closely related to members of the genus Methanosarcina, but clearly differed from all described species of this genus (95.6-97.6% of sequence similarity) . The level of DNA-DNA hybridization of strain ZS with Methanosarcina barkeri and Methanosarcina mazei was 15 and 31%, respectively . Based on the results of physiological and phylogenetic studies strain ZS can be assigned to a new species of the genus Methanasarcina . The name Methanosarcina lacustris sp . nov . is proposed . The type strain is ZS (= DSM 13486T, VKM B-2268).

J Pharm Belg, 2001 Nov-Dec, 56(6), 137 - 48
{Flouroquinolones of the third and fourth generations}; Rossi C et al.; Levofloxacin, levorotatory isomer of ofloxacine, is the only FQ3G on the belgian market since the middle of 2000 and is a little more efficient than the FQ2G against S . pneumoniae . The FQ4G are in vitro active against the common and atypic respiratory pathogenes and significantly more efficient against the Gram positive cocci, principally against S . pneumoniae . Their long duration of action allows one oral administration a day sufficient to obtain bactericidal levels in the serum . Their secondary effects are located principally at the level of digestive tract, neurologic system or cutaneous area . In the U.S.A., several FQ4G (clina-, grepa-, spar-, trova-floxacin) were withdrawn from use after observance of severe toxicity, while there were being administrated on a large scale . Furthermore, two of these "respiratory" FQ4G (gemi- and moxifloxacin) should reinforced our therapeutic armoury, moxifloxacin as of 2002 and gemifloxacin as of 2004 . The position of FQ3G and 4G are in discussion . In the case of pneumonia acquired in the community and confirmed by X-Ray, our recommendation is to administer a new FQ at the first choice if the patient is allergic to penicillin, debilitated or if a penicillin resistant S . pneumoniae is suspected (epidemiology, recent antibiotherapy) and at the second choice if it is resistance to penicilline or to macrolide, in case of atypic pneumonia . Today, the majority of the pneumonia acquired outside the hospital is empirically treated with a betalactam (oral or intravenous), with or without macrolide . The new FQ are important drugs . One must avoid overprescription which leads to bacterial resistance (especially S . pneumoniae).

Antonie Van Leeuwenhoek, 2001 Sep, 79(3-4), 235 - 45
Heterologous expression of the naphthocyclinone hydroxylase gene from Streptomyces arenae for production of novel hybrid polyketides; Brunke P et al.; Streptomyces arenae produces at least four different isochromanequinone antibiotics, the naphthocyclinones, of which the beta- and gamma-form are active against Gram-positive bacteria . The naphthocyclinone biosynthesis gene cluster was isolated from Streptomyces arenae DSM 40737 and by sequence analysis the minimal polyketide synthase genes and several genes encoding tailoring enzymes were identified . Southern blot analysis of the naphthocyclinone gene cluster indicated that a 3.5 kb BamHI fragment located approximately 9 kb downstream of the minimal PKS genes hybridizes to the schC hydroxylase DNA probe isolated from S . halstedii . Two complete and one incomplete open reading frames were identified on this fragment . Sequence analysis revealed strong homology to the genes of the actVA region of S . coelicolor, to several (suggested) hydroxylases and a putative FMN-dependent monooxygenase . The proposed hydroxylase, encoded by ncnH, could hydroxylate aloesaponarin II, a molecule that is produced by the actinorhodin minimal polyketide synthase in combination with the actinorhodin ketoreductase, aromatase and cyclase . Furthermore, this enzyme is capable of accepting additional polyketide core structures that contain a 5-hydroxy-1,4-naphthoquinone moiety as substrates which makes it an interesting tailoring enzyme for the modification of polyketide structures.

Salud Publica Mex, 2001 Nov-Dec, 43(6), 515 - 23
{Incidence of nosocomial bacteremia and pneumonia in pediatric unit}; Martinez-Aguilar G et al.; OBJECTIVE: To determine the incidence of catheter-related bacteremia and ventilator-associated pneumonia in children admitted to a secondary care hospital . MATERIAL AND METHODS: A prospective active surveillance system was conducted from January 1999 to June 2000, at the Hospital General of Instituto Mexicano del Seguro Social in Durango, Mexico . Daily visits to the pediatric ward were conducted to detect episodes of bacteremia and pneumonia, according to the Official Mexican Norm . Hospitalized patients under mechanical ventilation and/or with a central venous catheter, were followed from the first day of exposure, until a nosocomial infection was detected, or until the invasive device was removed . Blood and tracheal aspirate cultures were obtained from all exposed patients . Incidence rates with 95% confidence intervals were calculated for ventilator-associated pneumonia and bacteremia/sepsis per 1000 exposure days . Also, the monthly infection rate is presented for days of exposure, using statistical control graphs . RESULTS: A total of 47 episodes of bacteremia/sepsis and 44 of ventilator associated pneumonia were recorded . The incidence rate of pneumonia and bacteremia/sepsis was 28 and 26 cases respectively, per 1000 days of exposure to and invasive device . The gram-positive rods (61.11%) were more common than the gram negative rods (38.88%) . CONCLUSIONS: The most striking finding of this study was the higher incidence of these two nosocomial infections in children, as compared to that reported elsewhere . These findings call for preventive strategies and guidelines for handling intravenous catheters and mechanical ventilation in Mexico.

J Infect Chemother, 1999 Mar, 5(1), 40 - 45
Teicoplanin for skin and soft tissue infections: An open study and a randomized, comparative trial versus cefazolin; Stevens DL; An open trial and a multicenter, three-group, randomized trial versus cefazolin were performed to study the use of teicoplanin in the treatment of serious skin and soft tissue infections caused by Gram-positive bacteria . A total of 418 patients were entered into the randomized trial, 293 of whom were available for efficacy analysis, and 262 patients were entered in the open trial . The randomized trial had three arms: intramuscular (125 patients) vs intravenous (148 patients) teicoplanin vs cefazolin (145 patients) . In both trials teicoplanin was administered once daily, originally as 3 mg/kg per day, with the option of higher doses in the open trial . Cefazolin was given at a dose of 1.5-4 g/day, in three divided doses . In the randomized trial, teicoplanin and cefazolin showed similar overall efficacy . The higher dose of teicoplanin (6 mg/kg) was significantly more effective than the lower dose (3 mg/kg), particularly in patients with diabetes . In the open trial, teicoplanin had a clinical success rate of 93% . There was no significant difference in the incidence of adverse events between the cefazolin and teicoplanin groups . Outpatient ambulatory therapy was shown to be a practical method of administering teicoplanin . Once-daily dosing with teicoplanin may allow physicians to treat skin and soft tissue infections on a totally outpatient basis.

J Infect Chemother, 1999 Mar, 5(1), 32 - 39
Teicoplanin in the treatment of bone and joint infections: An open study; LeFrock J et al.; Osteomyelitis and septic arthritis caused by Gram-positive pathogens may require prolonged inpatient treatment . The glycopeptide antibiotic, teicoplanin, can be administered once daily to outpatients, and was assessed in a multicenter, open trial in patients with such infections . Patients with proven Gram-positive osteomyelitis or septic arthritis were treated with once-daily teicoplanin, 6-12 mg/kg per day, after three loading doses at intervals of 12 h, for 4-6 weeks . A total of 342 patients were recruited, of whom 220 were fully evaluable . Surgical procedures were performed in 82% of patients . Clinical success by the end of treatment was recorded in 81/90 patients (90%) with acute osteomyelitis, 70/79 patients (88.6%) with chronic osteomyelitis, and 42/51 patients (82.4%) with septic arthritis . Four patients with acute and 4 with chronic osteomyelitis and 5 patients with septic arthritis failed to respond to treatment . Relapse was known to have occurred in 10 patients with osteomyelitis and 4 with septic arthritis . Mean trough levels of teicoplanin reached during the first week of therapy were 10 mg/l (mean dose, 6 mg/kg) and 21 mg/l (mean dose, 12 mg/kg) . A mean of 75% of the treatment course was given at home . One or more adverse events were reported in 166/342 patients (48.5%), 119 (34.8%) of which were thought to be related to teicoplanin, and treatment was discontinued in 59 patients . Fever, chills, and rashes were the most common side-effects, but were usually mild . Teicoplanin was shown to be a cost-effective method of treatment of bone and joint infections caused by multiple-resistant Gram-positive pathogens.

Intensive Care Med, 2001 Nov, 27(11), 1770 - 5 Epub 2001 Oct 30.
Lack of association between the -260 C-->T promoter polymorphism of the endotoxin receptor CD14 gene and the CD14 density of unstimulated human monocytes and soluble CD14 plasma levels; Heesen M et al.; OBJECTIVE: CD14 is a receptor for endotoxin and binds components of Gram-positive and Gram-negative bacteria . CD14-bearing monocytes respond to stimulation with the increased synthesis and release of cytokines . The recently described -260 C-->T promoter polymorphism of the CD14 gene has been found to be related to a risk of myocardial infarction . This study evaluated the role of this polymorphism in the expression of monocyte and soluble CD14 . Moreover, the effect of the CD14 -260 genotypes for the ex vivo TNF-alpha response to endotoxin was analyzed in whole blood . PATIENTS AND PARTICIPANTS: Ninety-five healthy blood donors were studied . MEASUREMENTS AND RESULTS: CD14 -260 genotyping was performed by means of a real-time PCR with fluorescence labeled hybridization probes . CD14 expression on human monocytes (mCD14) was assessed by fluorescence-activated cell sorting analysis with anti-CD14 monoclonal antibodies . Plasma levels of soluble CD14 (sCD14) were measured by ELISA . The TNF-alpha synthesis was determined by chemiluminescence in whole blood after endotoxin stimulation . There were no differences in mCD14 density, sCD14 levels, or the tumor necrosis factor-alpha concentrations between individuals with the three different CD14 -260 genotypes CC, CT, and TT . CONCLUSIONS: The CD14 -260 polymorphism does not affect the CD14 expression of unstimulated circulating monocytes or soluble CD14 plasma levels.

Arch Microbiol, 2002 Feb, 177(2), 197 - 9 Epub 2001 Dec 07.
Hemagglutinin of unusual specificity from Helcococcus kunzii; Stavri H et al.; Helcococcus kunzii is a gram-positive, catalase-negative opportunist . The organism has been isolated from the lower extremities and breast masses of several patients . A clinical isolate of Helcococcus kunzii was shown to possess a hemagglutinin-lectin with a specificity for N-acetylglucosamine and lactose, two structurally unrelated carbohydrates . The lectin is sensitive to protease, heat and mutanolysin . Electron microscopy failed to reveal fimbriae or fibrillae, suggesting that the lectin is associated with peptidoglycan or the cytoplasmic membrane . It is likely that the lectin is involved in adhesion and colonization of H . kunzii.

Arch Microbiol, 2002 Feb, 177(2), 184 - 91 Epub 2001 Dec 07.
The cell wall of Micromonospora purpurea contains a high conductance channel; Kim BH et al.; In this communication it is demonstrated that the cell wall of the gram-positive bacterium Micromonospora purpurea contains a cell wall channel for the passage of hydrophilic solutes . The channel-forming protein was identified in sucrose step-density-gradient fractions of the cell envelope and in whole cell extracts using either organic solvent or detergent and the lipid bilayer technique . The fractions of the sucrose step-density centrifugation were assayed for NADH-oxidase activity and for the formation of ion-permeable channels in lipid bilayers . The highest NADH-oxidase activity and the highest channel-forming ability were found in different fractions . The cell wall fraction was identified by the presence of meso-diaminopimelic acid and contained an ion-permeable channel with the extremely high single-channel conductance of about 14 nS in 1 M KCl . The channel-forming unit was purified to homogeneity by FPLC on a HiTrap-Q column . It was identified as a heat- and SDS-resistant 200-kDa band on SDS-PAGE and formed the same general diffusion pores in lipid bilayer membranes as those formed by detergent extracts of the cell wall fraction of the sucrose step-density centrifugation . The channels were slightly selective for potassium ions over chloride, possibly caused by an excess of negative charges in or near the channel.

Shock, 2002 Jan, 17(1), 19 - 22
Delayed preconditioning induced by lipoteichoic acid from B . subtilis and S . aureus is not blocked by administration of 5-hydroxydecanoate; Zacharowski K et al.; Bacterial walls contain lipopolysaccharide (LPS), lipoteichoic acid (LTA), or peptidoglycan . Pretreatment of rats with low doses of LPS (from E . coli) or LTA (from S . aureus, a pathogenic gram-positive bacterium) for 16-24 h reduces myocardial infarct size caused by a subsequent period of myocardial ischemia-reperfusion . This phenomenon of enhanced tolerance to an ischemic insult has been termed delayed preconditioning (DP) . The aim of this study was to investigate whether LTA from B . subtilis (a nonpathogenic gram-positive bacterium) induces DP when administered 16 h before left anterior descending coronary artery (LAD) occlusion-reperfusion in the rat . Furthermore, we investigated whether the specific mitochondrial K(ATP) (mitoK(ATP)) channel inhibitor 5-hydroxydecanoate (5-HD, 5 mg/kg) blocks DP afforded by LTA of both strains of bacteria . Male Wistar rats were subjected to LAD occlusion-reperfusion (25-120 min) and infarct size was determined . In rats pretreated with saline (1 mL/kg i.p.), LAD occlusion-reperfusion resulted in an infarct size of 58% . Pretreatment of animals with LTA (S . aureus, 1 mg/kg i.p.) or LTA (B . subtilis, 1 mg/kg i.p.) reduced infarct size by 22% or 33%, respectively . Administration of 5-HD 10 min before LAD occlusion-reperfusion did not abolish DP afforded by LTA from S . aureus or B . subtilis, respectively . These results imply that late (after 16 h) opening of the mitoK(ATP) channel is not part of the signaling pathway of LTA-induced DP.

Curr Opin Crit Care, 2001 Oct, 7(5), 371 - 5
Toll receptors and sepsis; Read RC et al.; Toll-like receptors are a family of receptors that recognize components of bacteria and induce a proinflammatory response by cells, including macrophages and endothelial cells . Ten human Toll receptors differing in their specificity for microbial components have been cloned . They respond to various components, including lipopolysaccharide of Gram-negative bacteria, lipopeptides of Gram-positive cell walls, bacterial DNA, and flagella . Some Toll-like receptors require the cooperation of an adapter protein . Toll-like receptor 4 function requires the presence of the protein MD2 . Recently, it has been shown that Toll-like receptors function cooperatively to increase the specificity of response to a given microbe . Human polymorphisms of Toll-like receptor genes have been discovered and are associated with hyporesponsiveness to bacterial components.

J Biol Chem, 2002 Apr 19, 277(16), 13966 - 72 Epub 2002 Jan 22.
X-ray crystallographic structures of the Escherichia coli periplasmic protein FhuD bound to hydroxamate-type siderophores and the antibiotic albomycin; Clarke TE et al.; Siderophore-binding proteins play an essential role in the uptake of iron in many Gram-positive and Gram-negative bacteria . FhuD is an ATP-binding cassette-type (ABC-type) binding protein involved in the uptake of hydroxamate-type siderophores in Escherichia coli . Structures of FhuD complexed with the antibiotic albomycin, the fungal siderophore coprogen and the drug Desferal have been determined at high resolution by x-ray crystallography . FhuD has an unusual bilobal structure for a periplasmic ligand binding protein, with two mixed beta/alpha domains connected by a long alpha-helix . The binding site for hydroxamate-type ligands is composed of a shallow pocket that lies between these two domains . Recognition of siderophores primarily occurs through interactions between the iron-hydroxamate centers of each siderophore and the side chains of several key residues in the binding pocket . Rearrangements of side chains within the binding pocket accommodate the unique structural features of each siderophore . The backbones of the siderophores are not involved in any direct interactions with the protein, demonstrating how siderophores with considerable chemical and structural diversity can be bound by FhuD . For albomycin, which consists of an antibiotic group attached to a hydroxamate siderophore, electron density for the antibiotic portion was not observed . Therefore, this study provides a basis for the rational design of novel bacteriostatic agents, in the form of siderophore-antibiotic conjugates that can act as "Trojan horses," using the hydroxamate-type siderophore uptake system to actively deliver antibiotics directly into targeted pathogens.

Clin Infect Dis, 2002 Feb 15, 34(4), 482 - 92 Epub 2002 Jan 11.
Mechanisms of resistance to macrolides and lincosamides: nature of the resistance elements and their clinical implications; Leclercq R; Resistance to macrolides and lincosamides is increasingly reported in clinical isolates of gram-positive bacteria . The multiplicity of mechanisms of resistance, which include ribosomal modification, efflux of the antibiotic, and drug inactivation, results in a variety of phenotypes of resistance . There is controversy concerning the clinical relevance of in vitro macrolide resistance . Recent data, however, have shown that eradication of bacteria correlates with clinical outcome of acute otitis media in children and that macrolide therapy results in delayed eradication of macrolide-resistant pneumococci . These results support the need for in vitro detection of macrolide resistance and correct interpretation of susceptibility tests to guide therapy.

Intensive Care Med, 2001 Dec, 27(12), 1853 - 9 Epub 2001 Nov 10.
Activation of the fibrinolytic system and utilization of the coagulation inhibitors in sepsis: comparison with severe sepsis and septic shock; Mavrommatis AC et al.; OBJECTIVES: To determine whether the fibrinolytic system is activated and coagulation inhibitors are utilized in sepsis, to compare the findings detected in sepsis with those found in severe sepsis and septic shock, and to compare the role played by different infectious pathogens on fibrinolysis and coagulation inhibitors . DESIGN AND SETTING: Prospective study comparing patients with sepsis, severe sepsis, and septic shock and healthy volunteers in the general intensive care unit of a tertiary university hospital . PATIENTS: Eighty-two consecutive septic patients (47 with sepsis, 18 with severe sepsis, and 17 with septic shock), and 14 healthy volunteers (controls) . MEASUREMENTS AND RESULTS: After blood sampling we measured activation markers of fibrinolysis {plasmin/alpha(2)-antiplasmin complexes (PAP), complexes of tissue plasminogen activator/plasminogen activator inhibitor (tPA/PAI), fibrin(ogen) degradation products (FDPs), D-dimmers fibrin degradation products (D-d)}, the utilization marker of antithrombin III (ATIII) thrombin/antithrombin complexes (TAT), several factors of fibrinolysis {plasminogen, tissue plasminogen activator (tPA), plasminogen activator inhibitor 1 (PAI-1), alpha(2)-antiplasmin}, and the natural coagulation inhibitors {ATIII, protein C (PrC), protein S (PrS)} . In sepsis, PAP, FDPs, D-d, and TAT were increased to 439.8+/-32.35 microg/l, 57% positive, 49% positive, and 3.46+/-0.27 microg/l, respectively, compared with control subjects (205.57+/-28.58 microg/l, 0% positive, 7% positive, and 1.61+/-0.1 microg/l, respectively) . These markers further increased in severe sepsis and septic shock . With the exception of a decrease in ATIII and an increase in tPA and PAI-1, coagulation inhibitors and factors of fibrinolysis were not changed in sepsis . In severe sepsis and mainly in septic shock, coagulation inhibitors (ATIII, PrC) and plasminogen were markedly decreased, whereas tPA and PAI-1 were further increased . All changes were independent of the causative infectious pathogen . CONCLUSIONS: Fibrinolysis is strongly activated and ATIII is utilized in sepsis . These findings are further enhanced in severe sepsis and septic shock . In sepsis only ATIII is decreased . In contrast, in severe sepsis and mainly in septic shock plasminogen and the main coagulation inhibitors (i.e., ATIII, PrC) are depleted, indicating exhaustion of fibrinolysis and coagulation inhibitors . Finally, Gram-positive, Gram-negative and other micro-organisms produce identical impairment.

Biosci Biotechnol Biochem, 2001 Nov, 65(11), 2369 - 77
Effects of polysaccharides from rhizomes of Curcuma zedoaria on macrophage functions; Kim KI et al.; The effects of Curcuma zedoaria, which is used as a condiment, in perfumery, and as a medicine, on immune response were investigated by measuring macrophage-stimulating activity in macrophages and RAW 264.7 cells . In this study, CZ-1 and CZ-1-III, the fractions partially purified from C . zedoaria, had a strong, dose-dependent lysosomal enzyme activity . It was suggested that active portions of CZ-1-III were polysaccharides rather than proteins . Phagocytic activity increased as a similar pattern in both the gram-negative and gram-positive bacteria, time-dependently . It was demonstrated that CZ-1-III can augment the oxygen burst response but had an even higher activity in vivo than in vitro . Also a significant increase of H2O2, NO, and TNF-alpha production was observed . However, the production of TNF-alpha at the concentration of 1,000 microg/ml decreased . These data suggested that C . zedoaria had macrophage-stimulating activity and the possibility of being used as a biological response modifier.

Curr Opin Immunol, 2002 Feb, 14(1), 123 - 8
Scavenger receptors in innate immunity; Peiser L et al.; Scavenger receptors (SR) are expressed by myeloid cells (macrophages and dendritic cells) and certain endothelial cells . They play an important role in uptake and clearance of effete components, such as modified host molecules and apoptotic cells . They bind and internalise micro-organisms and their products including Gram-positive bacteria (lipoteichoic acid), Gram-negative bacteria (lipopolysaccharide), intracellular bacteria and CpG DNA . SR can alter cell morphology and their expression is affected by various cytokines . SR are involved in lipid metabolism and bind modified low-density lipoproteins.

Ophthalmol Clin North Am, 2001 Dec, 14(4), 611 - 24
Gentamicin and other antibiotic toxicitiy; Thomas T et al.; Antibiotics have the potential to cause significant ocular toxicity when they gain access to the inside of the eye . The aminoglycosides, in particular gentamicin, are the most toxic of the antibiotics commonly used in ophthalmology . Extreme caution should be used when administering a periocular injection of aminoglycoside for treatment or prophylaxis of infection . Intraocular injection of aminoglycoside for gram-negative coverage in endophthalmitis management has been replaced in most cases by ceftazidime . Ceftazidime provides excellent coverage against gram-negative bacteria with less potential for retinal toxicity at therapeutic dosages . Experimental and clinical studies have shown that intraocular vancomycin is safe and effective treatment against gram-positive organisms causing endophthalmitis . A combination of ceftazidime and vancomycin provides broad-spectrum coverage for virtually all bacteria causing endophthalmitis and is the current intraocular treatment of choice.

Med Oncol, 2001, 18(1), 51 - 7
Serum levels of IL-1 beta, sIL-2R, IL-6, IL-8, and TNF-alpha in febrile children with cancer and neutropenia; Soker M et al.; Serum levels of interleukin-1 beta (IL-1beta), soluble interleukin 2 receptors (sIL-2R), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-alpha) were measured to predict some characteristics of febrile episodes in children with cancer and neutropenia . Forty-eight episodes of febrile neutropenia were determined in 23 pediatric cancer patients, including 35 febrile episodes without identifiable source, 7 episodes of bacteremia due to Gram-negative organisms and 4 due to Gram-positive organisms, and 2 fungal infections . Interleukin-6, sIL-2R, and IL-8 levels were significantly higher at the beginning of the febrile episodes than those of controls (p < 0.001, p < 0.001, and p < 0.001) . Interleukin-6, slL-2R, and IL-8 levels were higher in patients with bacteremia due to Gram-negative organisms than in those with Gram-positive ones (p = 0.042, p = 0.006, and p = 0.023, respectively) . TNF-alpha and IL-1beta levels were similar in febrile episodes and controls (p > 0.05) . In conclusion, sIL-2R, IL-6, and IL-8 levels may be helpful in the prediction of infection in febrile cancer patients with neutropenia and measurements of IL-1beta and TNF-alpha were not useful for identifying the presence and the type of infection in febrile neutropenic episodes in children.

Auris Nasus Larynx, 2002 Jan, 29(1), 59 - 63
Contemporary trends in microbiology and antibiotic resistance in otolaryngology; Bhattacharyya N et al.; OBJECTIVE: to determine current microbiologic yields and characterize antibiotic resistance patterns for organisms cultured from head and neck infections . METHODS: results of out-patient cultures submitted from an academic otolaryngology practice over the period 1994-1998 were reviewed . A database was constructed, and culture results were analyzed for sites of infection, organism recovery rate, infecting microbiological agent, antibiotic resistance patterns, and 5-year trends . chi(2) analysis was used to determine associations between site of infection, infecting agent and antibiotic resistance over the years of the study . RESULTS: a total of 986 aerobic cultures were reviewed . The most common sites cultured were the nose/paranasal sinuses (469), throat (377), and ear (23) . In 465 (47.2%) cultures, normal flora or no growth occurred . Of the throat culture subset, 69.7% were negative, whereas 69.1% of sinonasal cultures recovered organisms . Gram positive cocci were the most commonly recovered organism type (30.9%), followed by Gram negative rods (GNR, 17.2%) . These rates remained relatively constant over the years of the study . GNR were found in 39.1 and 25.2% of ear and sinus cultures, respectively . Antibiotic sensitivities were conducted on 257 specimens (49.3% of positive cultures) . The average number of antibiotic resistances per bacteria remained stable at approximately 1.59 over the years (P=0.086, analysis of variance (ANOVA)) . Antibiotic resistance rates increased for clindamycin, cefazolin, and erythromycin over the years of the study (P<0.05, chi(2)) . Ciprofloxacin, gentamicin and trimethoprim-sulfamethoxazole resistance rates did not increase over the years of the study . Vancomycin resistance did not occur . CONCLUSIONS: as a significant proportion of head and neck cultures will be negative, their utility should be re-examined . GNR infections are more common in the head and neck than earlier thought . There is a trend towards increasing antibiotic resistance in head and neck infections, urging careful and appropriate use of antibiotics.

Med J Malaysia, 2001 Jun, 56(2), 201 - 6
Bacteriological study and its role in the management of open tibial fracture; Faisham WI et al.; Sixty percent of open fracture wounds are contaminated at the time of injury . Despite that, the necessity for sequential multiple cultures and sensitivity studies for open fractures and their interpretation are still controversial . Predebridement, intraoperative, postoperative swabs and swabs in established infection for culture and sensitivity study were taken in 33 open tibial fractures over a 6 months period . 39.3% of predebridement swabs grew bacteria with the majority yielding gram-positive organism . None of the patients developed infection with similar organisms . 24.2% of the postoperative swabs grew bacteria, of which 75% were gram-negative . 50% of the patients with positive postoperative swabs developed infection . Thus, the role of sequential multiple cultures and sensitivity studies are not helpful in management of open fracture.

Pharmacotherapy, 2001 Dec, 21(12), 1579 - 82
Gatifloxacin-associated acute hepatitis; Henann NE et al.; Gatifloxacin, a fluoroquinolone with extended gram-positive activity, has become extensively used in both the community and hospital environments . Unfortunately, concerns have been raised about the use of certain fluoroquinolones because of adverse drug reactions . A 44-year-old woman developed acute hepatitis while receiving gatifloxacin for chronic sinusitis . After 5 days of receiving antibiotics, the patient developed nausea, lethargy, and abdominal pain, all of which progressed over the next few days . Liver function tests were elevated, with bilirubin peaking at 9.4 mg/dl . The patient also became jaundiced . A percutaneous liver biopsy showed acute hepatitis with eosinophilic infiltrates consistent with drug-induced hepatitis . All other drugs and disease processes were ruled out as likely causes of the patient's hepatitis . Clinicians should be alerted to the possibility that hepatitis may occur with gatifloxacin administration.

Res Microbiol, 2001 Nov, 152(9), 811 - 22
Mercury resistance transposons of gram-negative environmental bacteria and their classification; Mindlin S et al.; A total of 29 mercury resistance transposons were isolated from mercury-resistant environmental strains of proteobacteria collected in different parts of Eurasia and the USA and tested for hybridization with probes specific for transposase genes of known mercury resistance transposons . 9 were related to Tn21 in this test, 12 were related to Tn5053, 4 to Tn5041 and 1 to Tn5044; three transposons were negative in this test . Restriction mapping and DNA sequencing revealed that 12 transposons were identical or nearly identical to their corresponding relatives while the rest showed varying divergence from their closest relatives . Most of these previously unknown transposons apparently arose as a result of homologous or site-specific recombination . One of these, Tn5046, was completely sequenced, and shown to be a chimera with the mer operon and the transposition module derived from the transposons related to Tn5041 and to Tn5044, respectively . Transposon Tn5070, showing no hybridization with the specific probes used in this study, was also completely sequenced . The transposition module of Tn5070 was most closely related to that of Tn3 while the mer operon was most closely related to that of plasmid pMERPH . The merR of Tn5070 is transcribed in the same direction as the mer structural genes, which is typical for mer operons of gram-positive bacteria . Our data suggest that environmental bacteria may harbor many not yet recognized mercury resistance transposons and warrant their further inventory.

Ann Oncol, 2001 Oct, 12(10), 1359 - 68
Reduction of chemotherapy-induced febrile leucopenia by prophylactic use of ciprofloxacin and roxithromycin in small-cell lung cancer patients: an EORTC double-blind placebo-controlled phase III study; Tjan-Heijnen VC et al.; BACKGROUND: CDE (cyclophosphamide, doxorubicin, etoposide) is one of the standard chemotherapy regimens in the treatment of small-cell lung cancer (SCLC), with myelosuppression as dose-limiting toxicity . In this trial the impact of prophylactic antibiotics on incidence of febrile leucopenia (FL) during chemotherapy for SCLC was evaluated . PATIENTS AND METHODS: Patients with chemo-naive SCLC were randomized to standard-dose CDE (C 1,000 mg/m2 day 1, D 45 mg/m2 day 1, E 100 mg/m2 days 1-3 . i.v., q 3 weeks, x5) or to intensified CDE chemotherapy (125% dose, q 2 weeks, x4, with filgrastim 5 microg/kg/day days 4-13) to assess the impact on survival (n = 240 patients) . Patients were also randomized to prophylactic antibiotics (ciprofloxacin 750 mg plus roxithromycin 150 mg, bid . days 4-13) or to placebo in a 2 x 2 factorial design (first 163 patients) . This manuscript focuses on the antibiotics question . RESULTS: The incidence of FL during the first cycle was 25% of patients in the placebo and 11% in the antibiotics arm (P = 0.010; 1-sided), with an overall incidence through all cycles of 43% vs . 24% respectively (P = 0.007; 1-sided) . There were less Gram-positive (12 vs . 4), Gram-negative (20 vs . 5) and clinically documented (38 vs . 15) infections in the antibiotics arm . The use of therapeutic antibiotics was reduced (P = 0.013; 1-sided), with less hospitalizations due to FL (31 vs . 17 patients, P = 0.013: 1-sided) . However, the overall number of days of hospitalization was not reduced (P = 0.05; 1-sided) . The number of infectious deaths was nil in the antibiotics vs . five (6%) in the placebo arm (P = 0.022; 2-sided) . CONCLUSIONS: Prophylactic ciprofloxacin plus roxithromycin during CDE chemotherapy reduced the incidence of FL, the number of infections, the use of therapeutic antibiotics and hospitalizations due to FL by approximately 50%, with reduced number of infectious deaths . For patients with similar risk for FL, the prophylactic use of antibiotics should be considered.

Hosp Med, 2001 Nov, 62(11), 682 - 6
Linezolid: a new antibiotic for gram-positive infections; Wilson P; Linezolid is a new antibiotic that can treat multiresistant gram-positive infections and can be given parenterally or by mouth . It is not related to any existing agent so cross resistance is unusual.

Int J Syst Evol Microbiol, 2001 Nov, 51(Pt 6), 1969 - 76
Microbacterium gubbeenense sp . nov., from the surface of a smear-ripened cheese; Brennan NM et al.; Phenotypic and phylogenetic studies were performed on 11 strains of a Microbacterium-like organism isolated from the surface of a smear-ripened cheese . The isolates were Gram-positive, catalase-positive, facultatively anaerobic, oxidase-negative, non-spore-forming, non-motile, small, slender rods and grew in 12% (w/v) NaCl . Chemotaxonomic investigation revealed that all the isolates belonged unambiguously to the genus Microbacterium . They contained type B1 peptidoglycans with L-lysine as the diamino acid and glycolyl acyl types; rhamnose and galactose were the cell wall sugars . The G+C content ranged from 69 to 72 mol% . The major menaquinones were MK-11 and MK-12 and the major fatty acids were anteiso C15:0 and C17:0 and iso C16:0 . Phylogenetic analysis of the 16S rRNA sequences of four isolates showed that they represented a new subline in the genus Microbacterium, with Microbacterium barkeri as their nearest phylogenetic neighbour . M . barkeri showed the highest sequence similarity to the isolates; however, DNA-DNA hybridization showed that the isolates had only 38% chromosomal similarity to M . barkeri . Based on the phylogenetic and phenotypic distinctiveness of the isolates, it is proposed that they be classified as a new Microbacterium species, for which the name Microbacterium gubbeenense sp . nov . is suggested . The type strain has been deposited as LMG S-19263T (= NCIMB 30129T) . The GenBank accession number for the 16S rDNA sequence of the type strain is AF263563.

Immunity, 2001 Dec, 15(6), 1027 - 38
Drosophila scavenger receptor CI is a pattern recognition receptor for bacteria; Ramet M et al.; One hallmark of innate immunity apparently conserved from primitive life forms through to humans is the ability of the host to recognize pathogen-associated molecular patterns (PAMPs) . Since macrophage pattern recognition receptors are not well defined in Drosophila, we set out to identify such receptors . Our findings reveal that Drosophila macrophages express multiple pattern recognition receptors and that the Drosophila scavenger receptor, dSR-CI, is one such receptor capable of recognizing both gram-negative and gram-positive bacteria, but not yeast . Our data indicate that scavenger receptor bacterial recognition is conserved from insects to humans and may represent one of the most primitive forms of microbial recognition.

Sci STKE . 2000 Aug 08;2000(44):RE1.
The interleukin-1 receptor/Toll-like receptor superfamily: signal transduction during inflammation and host defense; O'Neill LA; The signal transduction pathways activated by the proinflammatory cytokine interleukin-1 (IL-1) have been the focus of much attention because of the important role that IL-1 plays in inflammatory diseases . A number of proteins have been described that participate in the post-receptor activation of the transcription factor NF-kappaB and stress-activated protein kinases such as p38 mitogen-activated protein kinase (MAPK) . It has also emerged that the type I IL-1 receptor (termed IL-1RI) is a member of an expanding receptor superfamily . These related receptors all have sequence similarity in their cytosolic regions . The family includes the Drosophila melanogaster protein Toll, the IL-18 receptor (IL-18R), and the Toll-like receptors TLR-2 and TLR-4, which bind molecules from Gram-positive and Gram-negative bacteria, respectively . Because of the similarity of IL-1RI to Toll, the conserved sequence in the cytosolic region of these proteins has been termed the Toll-IL-1 receptor (TIR) domain . The same proteins activated during signaling by IL-1RI also participate in signaling by IL-18R and TLR-4 . The receptor superfamily is evolutionarily conserved; members occur in plants and insects and also function in host defense . The signaling proteins activated are also conserved across species . This receptor superfamily therefore represents an ancient signaling system that is a critical determinant of the innate immune and inflammatory responses.

FEMS Microbiol Lett, 2001 Dec 18, 205(2), 291 - 7
Oceanobacillus iheyensis gen . nov., sp . nov., a deep-sea extremely halotolerant and alkaliphilic species isolated from a depth of 1050 m on the Iheya Ridge; Lu J et al.; An extremely halotolerant and alkaliphilic bacterium was isolated previously from deep-sea sediment collected at a depth of 1050 m on the Iheya Ridge . The strain, designated HTE831 (JCM 11309, DSM 14371), was Gram-positive, strictly aerobic, rod-shaped, motile by peritrichous flagella, and spore-forming . Strain HTE831 grew at salinities of 0-21% (w/v) NaCl at pH 7.5 and 0-18% at pH 9.5 . The optimum concentration of NaCl for growth was 3% at both pH 7.5 and 9.5 . The G+C content of its DNA was 35.8%.Low level (12-30%) of DNA-DNA relatedness between strain HTE831 and the species of these genera was found, indicating that HTE831 could not be classified as a member of a new species belonging to known genera . Based on phylogenetic analysis using 16S rDNA sequencing, chemotaxonomy, and the physiology of strain HTE831, it is proposed that this organism is a member of a new species in a new genus, for which the name Oceanobacillus iheyensis is proposed.

Ann Agric Environ Med, 2001, 8(2), 177 - 85
Bioaerosol emissions from a suburban yard waste composting facility; Hryhorczuk D et al.; The purpose of this study was to assess worker and community exposure to bioaerosols emitted from a suburban yard waste composting facility in northern Illinois, USA . Characterization of on- and off-site viable bioaerosols was undertaken through a total of 288 on- and off-site Andersen samples conducted over 10 sampling days . A total of 46 dust samples and 38 Kramer-Collins spore samples were also collected in this period . Evaluation of the impact of the facility on community bioaerosol concentrations was undertaken by comparing on- and off-site measurements by sampling locations, wind direction, and site activity . On-site concentrations of total spores, Aspergillus/Penicillium spores, total bacteria, Gram-positive bacteria, Gram-negative bacteria, actinomycetes, total particulates, endotoxin, and beta-1,3 glucans were higher than off-site concentrations . Total fungal spores averaged 13,451 spores/m3 (range 5,223-26,067) on-site and 8,772 spores/m3 (range 243-18,276) off-site . Viable bacterial airborne concentrations (in cfu/m3) averaged 11,879 on-site (range 480-78,880) and 3,204 off-site (range 160-17,600) . Mean levels of endotoxins (in ng/m3) were 1.94 on-site (range 0.12-6.06) and 0.14 off-site (range 0.01-0.41) . Mean levels of beta-1,3 glucans (in ng/m3) were 2.17 on-site (range 0.12-14.45) and 0.24 off-site (range 0.01-0.78) . Mean total viable fungi, on the other hand, were higher off-site than on-site (8,651 vs 3,068 cfu/m3) . On-site concentrations of total bacteria, Gram-positive bacteria, Gram-negative bacteria, and actinomycetes demonstrated a statistically significant pattern of decreasing concentration with distance from pile and higher downwind vs upwind concentrations . Mean on-site concentrations of viable bacteria, viable fungi, and endotoxins were significantly higher during periods of activity compared to periods of no activity . Off-site concentrations of bacteria were also significantly higher during periods of activity compared to no activity . The highest concentrations of total particulates, endotoxin, and beta-1,3-glucans were observed in the personal samplers worn by workers at the facility . One personal sampler measured an Asp f1 exposure of 22.17 ng/m3 during turning activity . Peak exposures to several bioaerosol constituents were sufficiently high to warrant use of respirators by workers during periods of pile activity that lead to dust generation.

Biopharm Drug Dispos, 2001 Apr, 22(3), 91 - 7
Linezolid absolute bioavailability and the effect of food on oral bioavailability; Welshman IR et al.; Linezolid is a novel oxazolidinone antibiotic that has a spectrum of activity encompassing a variety of Gram-positive bacteria . The objectives of this study were twofold: (1) to compare the absorption of linezolid tablets given immediately following a high-fat meal with the absorption of tablets administered while fasting, and (2) to assess the bioavailability of a 375-mg oral dose given while fasting relative to a 375-mg dose of linezolid sterile solution given intravenously . Venous blood samples were taken over the 48 h following the single dose administration of both the oral and intravenous (IV) treatment . Samples were subsequently frozen for the determination of linezolid concentrations by HPLC . The only statistically significant difference between the fasted and the fed treatment was in peak plasma concentration, with the mean C(max) for fasted subjects being 23% greater than that for subjects after consumption of a high-fat meal . Comparable AUC(0-infinity) values were measured under both conditions, indicating that the overall extent of absorption is the same . Therefore, the difference in C(max), while statistically significant, should not affect the therapeutic efficacy of linezolid when it is administered with food . There were no statistically significant differences in AUC(0-infinity), CL or half-life between the fasted oral treatment and the intravenous treatment . As expected, C(max) was statistically different between the two treatments . However, the mean absolute bioavailability (F) of the tablet, using the IV sterile solution as the reference treatment, was 103% (+/-20%) .

Nat Immunol, 2002 Jan, 3(1), 91 - 7 Epub 2001 Dec 17.
Drosophila MyD88 is required for the response to fungal and Gram-positive bacterial infections; Tauszig-Delamasure S et al.; We report here the identification and functional characterization of DmMyD88, a gene encoding the Drosophila homolog of mammalian MyD88 . DmMyD88 combines a Toll-IL-1R homology (TIR) domain and a death domain . Overexpression of DmMyD88 was sufficient to induce expression of the antifungal peptide Drosomycin, and induction of Drosomycin was markedly reduced in DmMyD88-mutant flies . DmMyD88 interacted with Toll through its TIR domain and required the death domain proteins Tube and Pelle to activate expression of Drs, which encodes Drosomycin . DmMyD88-mutant flies were highly susceptible to infection by fungi and Gram-positive bacteria, but resisted Gram-negative bacterial infection much as did wild-type flies . Phenotypic comparison of DmMyD88-mutant flies and MyD88-deficient mice showed essential differences in the control of Gram-negative infection in insects and mammals.

Nature, 2001 Dec 13, 414(6865), 756 - 9
Drosophila Toll is activated by Gram-positive bacteria through a circulating peptidoglycan recognition protein; Michel T et al.; Microbial infection activates two distinct intracellular signalling cascades in the immune-responsive fat body of Drosophila . Gram-positive bacteria and fungi predominantly induce the Toll signalling pathway, whereas Gram-negative bacteria activate the Imd pathway . Loss-of-function mutants in either pathway reduce the resistance to corresponding infections . Genetic screens have identified a range of genes involved in these intracellular signalling cascades, but how they are activated by microbial infection is largely unknown . Activation of the transmembrane receptor Toll requires a proteolytically cleaved form of an extracellular cytokine-like polypeptide, Spatzle, suggesting that Toll does not itself function as a bona fide recognition receptor of microbial patterns . This is in apparent contrast with the mammalian Toll-like receptors and raises the question of which host molecules actually recognize microbial patterns to activate Toll through Spatzle . Here we present a mutation that blocks Toll activation by Gram-positive bacteria and significantly decreases resistance to this type of infection . The mutation semmelweis (seml) inactivates the gene encoding a peptidoglycan recognition protein (PGRP-SA) . Interestingly, seml does not affect Toll activation by fungal infection, indicating the existence of a distinct recognition system for fungi to activate the Toll pathway.

Proc Natl Acad Sci U S A, 2001 Dec 18, 98(26), 15119 - 24 Epub 2001 Dec 11.
A genome-wide analysis of immune responses in Drosophila; Irving P et al.; Oligonucleotide DNA microarrays were used for a genome-wide analysis of immune-challenged Drosophila infected with Gram-positive or Gram-negative bacteria, or with fungi . Aside from the expression of an established set of immune defense genes, a significant number of previously unseen immune-induced genes were found . Genes of particular interest include corin- and Stubble-like genes, both of which have a type II transmembrane domain; easter- and snake-like genes, which may fulfil the roles of easter and snake in the Toll pathway; and a masquerade-like gene, potentially involved in enzyme regulation . The microarray data has also helped to greatly reduce the number of target genes in large gene groups, such as the proteases, helping to direct the choices for future mutant studies . Many of the up-regulated genes fit into the current conceptual framework of host defense, whereas others, including the substantial number of genes with unknown functions, offer new avenues for research.

Mol Microbiol, 2001 Nov, 42(4), 1007 - 20
Defining the disulphide stress response in Streptomyces coelicolor A3(2): identification of the sigmaR regulon; Paget MS et al.; In the Gram-positive, antibiotic-producing bacterium Streptomyces coelicolor A3(2), the thiol-disulphide status of the hyphae is controlled by a novel regulatory system consisting of a sigma factor, sigmaR, and its cognate anti-sigma factor, RsrA . Oxidative stress induces intramolecular disulphide bond formation in RsrA, which causes it to lose affinity for sigmaR, thereby releasing sigmaR to activate transcription of the thioredoxin operon, trxBA . Here, we exploit a preliminary consensus sequence for sigmaR target promoters to identify 27 new sigmaR target genes and operons, thereby defining the global response to disulphide stress in this organism . Target genes related to thiol metabolism encode a second thioredoxin (TrxC), a glutaredoxin-like protein and enzymes involved in the biosynthesis of the low-molecular-weight thiol-containing compounds cysteine and molybdopterin . In addition, the level of the major actinomycete thiol buffer, mycothiol, was fourfold lower in a sigR null mutant, although no candidate mycothiol biosynthetic genes were identified among the sigmaR targets . Three sigmaR target genes encode ribosome-associated products (ribosomal subunit L31, ppGpp synthetase and tmRNA), suggesting that the translational machinery is modified by disulphide stress . The product of another sigmaR target gene was found to be a novel RNA polymerase-associated protein, RbpA, suggesting that the transcriptional machinery may also be modified in response to disulphide stress . We present DNA sequence evidence that many of the targets identified in S . coelicolor are also under the control of the sigmaR homologue in the actinomycete pathogen Mycobacterium tuberculosis.

J Mol Biol, 2001 Dec 7, 314(4), 671 - 81
Dimer stabilization upon activation of the transcriptional antiterminator LicT; Declerck N et al.; LicT belongs to the BglG/SacY family of transcriptional antiterminators that induce the expression of sugar metabolizing operons in Gram positive and Gram negative bacteria . These proteins contain a N-terminal RNA-binding domain and a regulatory domain called PRD which is phosphorylated on conserved histidine residues by components of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) . Although it is now well established that phosphorylation of PRD-containing transcriptional regulators tunes their functional response, the molecular and structural basis of the regulation mechanism remain largely unknown.A constitutively active LicT variant has been obtained by introducing aspartic acid in replacement of His207 and His269, the two phosphorylatable residues of the PRD2 regulatory sub-domain . Here, the functional and structural consequences of these activating mutations have been evaluated in vitro using various techniques including surface plasmon resonance, limited proteolysis, analytical centrifugation and X-ray scattering . Comparison with the native, unphosphorylated form shows that the activating mutations enhance the RNA-binding activity and induce tertiary and quaternary structural changes . Both mutant and native LicT form dimers in solution but the native dimer exhibits a less stable and more open conformation than the activated mutant form . Examination of the recently determined crystal structure of mutant LicT regulatory domain suggests that dimer stabilization is accomplished through salt-bridge formation at the PRD2:PRD2 interface, resulting in domain motion and dimer closure propagating the stabilizing effect from the protein C-terminal end to the N-terminal effector domain . These results suggest that LicT activation arises from a conformational switch inducing long range rearrangement of the dimer interaction surface, rather than from an oligomerization switch converting an inactive monomer into an active dimer .

J Biol Chem, 2002 Mar 1, 277(9), 6830 - 7 Epub 2001 Nov 27.
Surfactant protein A inhibits peptidoglycan-induced tumor necrosis factor-alpha secretion in U937 cells and alveolar macrophages by direct interaction with toll-like receptor 2; Murakami S et al.; Pulmonary surfactant protein A (SP-A) plays an important role in modulation of the innate immune system of the lung . Peptidoglycan (PGN), a cell wall component of Gram-positive bacteria, is known to elicit excessive proinflammatory cytokine production from immune cells . In this study we investigated whether SP-A interacts with PGN and alters PGN-elicited cellular responses . Binding studies demonstrate that PGN is not a ligand for SP-A . However, SP-A significantly reduced PGN-elicited tumor necrosis factor alpha (TNF-alpha) secretion by U937 cells and rat alveolar macrophages . The inhibitory effect on TNF-alpha secretion was dependent upon SP-A concentrations in physiological range . Coincubation of SP-A and PGN with human embryonic kidney 293 cells that had been transiently transfected with the cDNA of Toll-like receptor 2 (TLR2), a cell signaling receptor for PGN, significantly attenuated PGN-induced nuclear factor-kappaB activity . SP-A directly bound to a soluble form of the recombinant extracellular TLR2 domain (sTLR2) . Coincubation of sTLR2 with SP-A significantly reduced the binding of sTLR2 to PGN . These results indicate that the direct interaction of SP-A with TLR2 alters PGN-induced cell signaling . We propose that SP-A modulates inflammatory responses against the bacterial components by interactions with pattern-recognition receptors.

Rev Med Brux, 2001 Oct, 22(5), 443 - 56
{Third and fourth generation fluoroquinolones}; Rossi C et al.; Levofloxacin, levorotatory isomer of ofloxacine, is the only FQ3G on the belgian market since the middle of 2000 and is a little more efficient than the FQ2G against S . pneumoniae . The FQ4G are in vitro active against the common and atypic respiratory pathogenes and significantly more efficient against the Gram positive cocci, principally against S . pneumoniae . Their long duration of action allows one oral administration a day sufficient to obtain bactericidal levels in the serum . Their secondary effects are located principally at the level of digestive tract, neurologic system or cutaneous area . In the U.S.A., several FQ4G (clina-, grepa-, spar-, trova-floxacin) were withdrawn from use after observance of severe toxicity, while there were being administrated on a large scale . Furthermore, two of these "respiratory" FQ4G (gemi- and moxi-floxacin) should reinforced our therapeutic armoury, moxifloxacin as of 2002 and gemifloxacin as of 2004 . The position of FQ3G and 4G are in discussion . In the case of pneumonia acquired in the community and confirmed by X-Ray, our recommendation is to administer a new FQ at the first choice if the patient is allergic to penicillin, debilitated or if a penicillin resistant S . pneumoniae suspected (epidemiology, recent antibiotherapy) and at the second choice if it is resistance to penicillin or to macrolide, in case of atypic pneumonia . Today, the majority of the pneumonia acquired outside the hospital is empirically treated with a betalactam (oral or intravenous), with or without macrolide . The new FQ are important drugs . One must avoid overprescription which leads to bacterial resistance (especially S . pneumoniae).

Br J Haematol, 2001 Oct, 115(1), 53 - 7
Procalcitonin-reduced sensitivity and specificity in heavily leucopenic and immunosuppressed patients; Svaldi M et al.; Procalcitonin (PCT) has proven to be a very sensitive marker of sepsis for non-leucopenic patients . Little is known about its relevance in immunosuppressed and leucopenic adults . Four hundred and seventy-five PCT determinations were carried out in 73 haematological patients: on 221 occasions the white blood cell (WBC) count was < 1.0 x 10(9)/l and on 239 occasions it was > 1.0 x 10(9)/l leucocytes . Patients were classified as: non-systemic infected controls (n = 280), patients with bacteraemia (n = 32), sepsis (n = 30), severe sepsis (n = 3), septic shock (n = 3) and systemic inflammatory response syndrome (SIRS) (n = 62) . When the WBC count was > 1.0 x 10(9)/l, gram-negative bacteria induced higher PCT levels (median 9.4 ng/ml) than gram-positives (median 1.4 ng/ml) . In cases with a WBC < 1.0 x 10(9)/l, PCT levels were similar for gram-negative and gram-positive bacteria (1.1 ng/ml versus 0.85 ng/ml) . Regardless of the leucocyte count, the median PCT level in bacteraemia cases always remained < 0.5 ng/ml . In heavily leucopenic situations, PCT levels were never > 2 ng/ml even in the sepsis and severe sepsis/septic shock groups, whereas a WBC count > 1.0 x 10(9)/l resulted in median PCT values of 4.1 ng/ml and 45 ng/ml respectively . The positive predictive value for sepsis (cut-off 2 ng/ml) was 93% in cases of WBC count > 1.0 x 10(9)/l, but only 66% in leucopenic conditions . The negative predictive value (cut-off 0.5 ng/ml) was 90% when the WBC count was > 1.0 x 10(9)/l and 63% in leucopenic conditions . Procalcitonin is an excellent sepsis marker with a high positive- and negative-predictive value in patients with WBC count > 1.0 x 10(9)/l, but it does not work satisfactorily below this leucocyte count.

Medicina (B Aires), 2001, 61(5 Pt 1), 577 - 80
{Ovarian abscess due to Actinomyces sp . in absence of an intrauterine contraceptive device}; Burlando SC et al.; The disease caused by Actinomyces spp . is often of difficult diagnosis . Actinomyces spp . are anaerobic or microaerophilic non-spore-forming gram-positive rods that may reach, occasionally, the normal female genital tract . IUD and pessaries facilitate the access of the microorganisms to the pelvis . We report an unusual case of ovarian infection by Actinomyces sp . in a 41 year-old female without IUD, admitted at the Institute in November 1998, with persistent fever . She had had an early menopause 3 years before, and had received hormonal replacement therapy . Usual and unusual infections were discarded by microbiological and serologic studies . Abdominal ultrasonography showed a slight left pyelocalycial dilatation and a simple cyst in the left ovary; heart ultrasonography was normal . Gynecological examination showed an enlarged uterus, similar to an 8 week pregnancy, painless, and fixed anexial masses . The transvaginal ultrasonography showed uterine myomas, one of them of 42 mm in the isthmus region, large ovaries, cystic, with acoustic shadows, and the left one with a septum . The preoperative diagnosis was infected bilateral cystic teratoma . The procedure was an exploratory laparotomy, followed by a bilateral salpingo-oophorectomy . The specimen studies showed an endometrioma with calcium deposits in the wall of the right ovary, and an abscess in the left ovary, also with calcification of the wall . The sample from the left abscess developed Actinomyces sp . After surgery, and treatment with penicillin, the fever disappeared . It is important to remark that the ovarian infection by Actinomyces sp . can also occur in patients without an IUD or a pessary; it might cause anexial images that can be interpreted as a tumour, inducing to erroneous diagnosis and treatment.

Eur J Cancer, 2001 Dec, 37(18), 2413 - 9
Incidence of septicaemias and invasive mycoses in children undergoing treatment for solid tumours: a 12-year experience at a single Italian institution; Haupt R et al.; We carried out a retrospective study on the infection rate--in episodes per 100 person months at risk (p/m/r)--of septicaemia and invasive mycoses in children with solid tumours treated at a single institution between 1985 and 1996 . Among 982 patients, accounting for 8108 p/m/r, 257 infectious episodes were documented, for an infection rate of 3.2 . The infection rate for "intensive" treatment was greater than that for "less intensive" treatments, 3.7 compared with 0.5, respectively; P<0.001 . 58% of infectious episodes were associated with neutropenia, 22% were megatherapy-related, and 39% were related to central venous catheter (CVC), while in 13% of the episodes no risk factor was identified . Of the episodes, single organism Gram-positive bacteraemias accounted for 62%, single organism Gram-negative for 23%, multiple organism bacteraemias for 7%, invasive mycoses for 4%, and isolated fungaemias for 4% . The infection rate for Gram-positive organisms decreased significantly over time (-5.9% per year; P<0.01), but increased for the Gram-negative organisms (+3.4% per year; P=0.4) . This study demonstrates that the risk of bacteraemia increases in parallel with the treatment intensity, and that a considerable number of children with solid tumours develop bacteraemia in the absence of an identifiable risk factor.

J Endotoxin Res, 2001, 7(4), 310 - 4
IC14, a CD14 specific monoclonal antibody, is a potential treatment for patients with severe sepsis; Axtelle T et al.; CD14 is a pattern recognition receptor for the bacterial cell wall components from Gram-positive and Gram-negative bacteria as well as mycobacteria . Binding of lipopolysaccharide (LPS) or other cell wall constituents to CD14 initiates signal transduction through the Toll-like receptors resulting in the release of pro-inflammatory cytokines and the initiation of the systemic inflammatory response . In rabbits and non-human primates, CD14 specific antibodies were shown to attenuate responses to LPS or Escherichia coli challenge including pro-inflammatory cytokine release, acute lung injury, hypotension and changes in lung, liver, spleen and adrenal gland morphology . In healthy human subjects, single doses of a chimeric CD14 antibody (IC14) have been shown to be well tolerated and result in a dose-related degree of saturation of CD14 receptors on monocytes and granulocytes . Pretreatment of healthy subjects with IC14 2 h prior to LPS resulted in an attenuation of the LPS-induced fever, clinical symptoms, and leukocyte activation and degranulation . IC14 inhibited the release of TNF-alpha, IL-6, and IL-10 and delayed the release of sTNFR(I) and IL-1ra . Further studies are in progress to characterize the safety and clinical pharmacology of IC14 in patients with severe sepsis.

J Am Chem Soc, 2001 Nov 28, 123(47), 11638 - 43
Lipid II: total synthesis of the bacterial cell wall precursor and utilization as a substrate for glycosyltransfer and transpeptidation by penicillin binding protein (PBP) 1b of Escherichia coli; Schwartz B et al.; An essential feature in the life cycle of both gram positive and gram negative bacteria is the production of new cell wall . Also known as murein, the cell wall is a two-dimensional polymer, consisting of a linear, repeating N-acetylmuramic acid (MurNAc) and N-acetylglucosamine (GlcNAc) motif, cross-linked via peptides appended to MurNAc . The final steps in the maturation of murein are catalyzed by a single, bifunctional enzyme, known as a high MW, class A penicillin binding protein (PBP) . PBPs catalyze polymerization of the sugar units (glycosyltransfer), as well as peptide cross-linking (transpeptidation) utilizing Lipid II as substrate . Detailed enzymology on this enzyme has been limited, due to difficulties in obtaining sufficient amounts of Lipid II, as well as the availability of a convenient and informative assay . We report the total chemical synthesis of Lipid II, as well as the development of an appropriate assay system and the observation of both catalytic transformations.

J Microbiol Methods, 2001 Dec, 47(3), 369 - 71
Use of magnetic beads for Gram staining of bacteria in aqueous suspension; Yazdankhah SP et al.; A Gram staining technique was developed using monodisperse magnetic beads in concentrating bacteria in suspension for downstream application . The technique does not require heat fixation of organisms, electrical power, or a microscope . Gram-negative and Gram-positive bacteria were identified macroscopically based on the colour of the suspension . The bacteria concentrated on magnetic beads may also be identified microscopically.

J Antibiot (Tokyo), 2001 Sep, 54(9), 730 - 6
Bagremycin A and B, novel antibiotics from streptomyces sp . Tü 4128; Bertasso M et al.; Two novel secondary metabolites, bagremycin A (2) and B (3), were detected in the culture filtrate of Streptomyces sp . Tu 4128 by HPLC-diode-array screening . They are phenol esters of 3-amino-4-hydroxybenzoic acid with a derivative of p-coumaric acid and show a moderate activity against gram-positive bacteria and some fungi.

J Neurochem, 2001 Nov, 79(3), 648 - 57
Circulating cell wall components derived from gram-negative, not gram-positive, bacteria cause a profound induction of the gene-encoding Toll-like receptor 2 in the CNS; Laflamme N et al.; The recent characterization of human homologs of Toll may be the missing link for the transduction events leading to nuclear factor-kappaB (NF-kappaB) activity and proinflammatory gene transcription during innate immune response . Mammalian cells may express as many as 10 distinct Toll-like receptors (TLRs), although TLR2 is a key receptor for recognizing cell wall components of Gram-positive bacteria . The present study investigated the effects of circulating bacterial cell wall components on the expression of the gene-encoding TLR2 across the mouse brain . Surprisingly, while Gram-negative components caused a robust increase in TLR2 transcription within the cerebral tissue, peptidoglycan (PGN) and lipoteichoic acid (LTA), either alone or combined, failed to modulate the receptor transcript . Indeed, the mRNA levels for TLR2 in the choroid plexus and few other regions of the brain remained similar between vehicle-, LTA-, PGN-, and LTA/PGN-administered mice at all the times evaluated (i.e . 30 min to 24 h post-intraperitoneal injection) . This contrasts with the profound de novo expression of TLR2 following a single systemic injection of the lipopolysaccharide (LPS) . The signal was first detected in regions devoid of blood-brain barrier and few blood vessels and microcapillaries . A second wave of TLR2 expression was also detected from these structures to their surrounding parenchymal cells that stained for a microglial marker iba1 . The rapid induction of IkappaBalpha (index of NF-kappaB activity) and up-regulation of the adaptor protein MyD88 suggest that LPS-induced TLR2 transcription may be dependent on the NF-kappaB pathway . These data provide the evidence that TLR2 is not only present in the brain, but its encoding gene is regulated by cell wall components derived from Gram-negative, not Gram-positive, bacteria . The robust wave of TLR2-expressing microglial cells may have a determinant impact on the innate immune response that occurs in the brain during systemic infection by Gram-negative, not Gram-positive, bacteria.

Crit Care Med, 2001 Nov, 29(11), 2075 - 80
Glutamine administration reduces Gram-negative bacteremia in severely burned patients: a prospective, randomized, double-blind trial versus isonitrogenous control; Wischmeyer PE et al.; OBJECTIVE: To determine the effect of intravenous glutamine supplementation vs . an isonitrogenous control on infectious morbidity in severely burned patients . Previous clinical studies in seriously ill patients suggest a beneficial effect of glutamine on infectious morbidity, but no trials have examined possible clinical benefits in severely burned patients . DESIGN: Prospective, double-blind, randomized trial . SETTING: Burn intensive care unit of a university hospital . PATIENTS: Twenty-six severe burn patients with total burn surface area of 25% to 90% and presence of full-thickness burns . Patients were evaluated for occurrence of bacteremia and antibiotic use during the first 30 days of their burn unit admission . Nutritional status and overall inflammation were also measured . INTERVENTION: Either intravenous glutamine or an isonitrogenous control amino acid solution was administered as a continuous infusion during burn intensive care unit stay . MEASUREMENTS AND MAIN RESULTS: The incidence of Gram-negative bacteremia was significantly reduced in the glutamine-supplemented group (8%) vs . control (43%; p <.04) . No difference was seen in the incidence of Gram-positive bacteremia or fungemia . Average number of positive blood cultures, antibiotic usage, and mortality rates also were reduced but did not reach statistical significance . Significant improvements in serum transferrin and prealbumin were observed in glutamine-supplemented patients at 14 days after burn injury (p <.01 and.04, respectively) . C-reactive protein was also significantly reduced at 14 days after burn injury in the glutamine group (p <.01) . CONCLUSIONS: Significantly fewer bacteremic episodes with Gram-negative organisms occurred in the glutamine-supplemented patients . Glutamine supplementation improved measures of nutrition and decreased measures of overall inflammation . In addition, a trend toward lower mortality rate, decreased overall bacteremia incidence, and antibiotic usage in the glutamine group was observed . Glutamine's beneficial effects may be a result of improved gut integrity or immune function, but the precise mechanism of glutamine's protection is unknown.

Leuk Lymphoma, 2001 Jul, 42(3), 379 - 86
A randomized prospective multicentre trial of cefpirome versus piperacillin-tazobactam in febrile neutropenia; Bauduer F et al.; Fever is frequently the only clinical sign of infection in patients with chemo-induced neutropenia . In this setting, empirical administration of broad spectrum antibiotics must be rapid . The aim of this work was to compare, for the first time, cefpirome (CPO) and piperacillin-tazobactam (PT) in a large randomized trial . Two hundred-eight febrile neutropenic episodes (FNE) (> or = 38.5 degrees C and ANC < or = 0.5 giga/l) were treated by randomization, as first line therapy, using either CPO 2 g x 2/day (105 cases) or PT 4 g x 3/day (103 cases), alone (CPO: 15/PT: 15), or plus aminoglycoside (165 cases, CPO: 82/PT: 83) or quinolone (CPO: 2/PT: 2) . There were 131 men and 77 women aged between 17 and 83 years (median: 49) who received chemotherapy (n = 160) or allogeneic (n = 10) or autologous (n = 38) stem cell transplantations . Underlying diseases were: acute leukemia (n = 131), lymphoma (n = 33), myeloma (n = 16), solid tumor (n = 8), myeloproliferative disorder (n = 9), chronic lymphoid leukemia (n = 5), aplastic anemia (n = 3), myelodysplasia (n = 3) . Distribution of age, neutropenia duration (median: 17 days), underlying disease, and protocol therapy duration (median: 11 days) was comparable in both arms . A microbiologically documented infection (MDI) was evidenced in 57 cases (27%) . Bacteria were isolated from blood cultures in 54 cases (Gram positive: 32 cases) . Their in vitro susceptibility rates to CPO and PT were not different . Two days after antibiotics initiation, clinical (fever disappearance) and microbiological (culture negativation) success rates (SR) were 62% for CPO versus 61% for PT and 50% versus 55% respectively in case of MDI (p = 0.89) . Two deaths and 77 failures were registered . At the end of protocol, SR (no antibiotic change/absence of superinfection) was 59% with CPO versus 50% with PT (p = 0.27) and 53% versus 40% respectively in the 151 cases with neutropenia > or = 10 days (p = 0.17) . The occurrence of side effects was similar in both arms . In our hands, the efficacy of CPO and PT was comparable for treating FNE.

Shock, 2001 Nov, 16(5), 344 - 8
Enhanced expression of Fc alpha receptor I on blood phagocytes of patients with gram-negative bacteremia is associated with tyrosine phosphorylation of the FcR-gamma subunit; Chiamolera M et al.; Sepsis caused by gram-negative bacteria is a common finding having high incidence and mortality . Fc alpha RI (CD89), a receptor for immunoglobulin A (IgA), has been shown to mediate bacterial phagocytosis, which might play a role in the pathogenesis of sepsis . In this study the expression and function of Fc alpha RI were analyzed on blood monocytes and neutrophils of patients with bacteremia . We found a marked increased in expression of the alpha- and gamma-subunits of the Fc alpha RI on both types of cells in patients with gram-negative bacteremia, but not in patients with gram-positive bacteremia . This increase was independent of serum IgA levels . Fc alpha RI M(r) was lower on cells from gram-negative patients than on cells from controls (50-65 kDa versus 55-75 kDa), despite a similar 32-kDa backbone, indicating altered glycosylation . Increased levels of Fc alpha RI on blood phagocytes correlated with enhanced serum IL-6 levels, but not with IFN gamma or TNF-alpha . FcR-gamma chain associated with Fc alpha RI was phosphorylated in patients neutrophils, indicating functional engagement of this receptor during gram-negative sepsis . Increased expression and activation of Fc alpha RI-gamma 2 complexes following gram-negative infections suggests its involvement in host defense against bacteria.

J Bacteriol, 2001 Dec, 183(23), 6727 - 32
Twin-arginine translocation pathway in Streptomyces lividans; Schaerlaekens K et al.; The recently discovered bacterial twin-arginine translocation (Tat) pathway was investigated in Streptomyces lividans, a gram-positive organism with a high secretion capacity . The presence of one tatC and two hcf106 homologs in the S . lividans genome together with the several precursor proteins with a twin-arginine motif in their signal peptide suggested the presence of the twin-arginine translocation pathway in the S . lividans secretome . To demonstrate its functionality, a tatC deletion mutant was constructed . This mutation impaired the translocation of the Streptomyces antibioticus tyrosinase, a protein that forms a complex with its transactivator protein before export . Also the chimeric construct pre-TorA-23K, known to be exclusively secreted via the Tat pathway in Escherichia coli, could be translocated in wild-type S . lividans but not in the tatC mutant . In contrast, the secretion of the Sec-dependent S . lividans subtilisin inhibitor was not affected . This study therefore demonstrates that also in general in Streptomyces spp . the Tat pathway is functional . Moreover, this Tat pathway can translocate folded proteins, and the E . coli TorA signal peptide can direct Tat-dependent transport in S . lividans.

J Exp Med, 2001 Nov 5, 194(9), 1231 - 42
Protein kinase Cepsilon is required for macrophage activation and defense against bacterial infection; Castrillo A et al.; To assess directly the role of protein kinase C (PKC)epsilon in the immune system, we generated mice that carried a homozygous disruption of the PKCepsilon locus . PKCepsilon(-/-) animals appeared normal and were generally healthy, although female mice frequently developed a bacterial infection of the uterus . Macrophages from PKCepsilon(-/-) animals demonstrated a severely attenuated response to lipopolysaccharide (LPS) and interferon (IFN)gamma, characterized by a dramatic reduction in the generation of NO, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-1beta . Further analysis revealed that LPS-stimulated macrophages from PKCepsilon(-/-) mice were deficient in the induction of nitric oxide synthase (NOS)-2, demonstrating a decrease in the activation of IkappaB kinase, a reduction in IkappaB degradation, and a decrease in nuclear factor (NF)kappaB nuclear translocation . After intravenous administration of Gram-negative or Gram-positive bacteria, PKCepsilon(-/-) mice demonstrated a significantly decreased period of survival . This study provides direct evidence that PKCepsilon is critically involved at an early stage of LPS-mediated signaling in activated macrophages . Furthermore, we demonstrate that in the absence of PKCepsilon, host defense against bacterial infection is severely compromised, resulting in an increased incidence of mortality.

Nat Struct Biol, 2001 Dec, 8(12), 1020 - 4
A novel solenoid fold in the cell wall anchoring domain of the pneumococcal virulence factor LytA; Fernandez-Tornero C et al.; Choline binding proteins are virulence determinants present in several Gram-positive bacteria . Because anchorage of these proteins to the cell wall through their choline binding domain is essential for bacterial virulence, their release from the cell surface is considered a powerful target for a weapon against these pathogens . The first crystal structure of a choline binding domain, from the toxin-releasing enzyme pneumococcal major autolysin (LytA), reveals a novel solenoid fold consisting exclusively of beta-hairpins that stack to form a left-handed superhelix . This unique structure is maintained by choline molecules at the hydrophobic interface of consecutive hairpins and may be present in other choline binding proteins that share high homology to the repeated motif of the domain.

Appl Microbiol Biotechnol, 2001 Oct, 57(1-2), 257 - 61
Chromate reduction and 16S rRNA identification of bacteria isolated from a Cr(VI)-contaminated site; Pattanapipitpaisal P et al.; A gram-positive, hexavalent chromium {chromate: Cr(VI)}-tolerant bacterium, isolated from tannery waste from Pakistan, was identified as a Microbacterium sp . by 16S rRNA gene sequence homology . The strain (designated as MP30) reduced toxic Cr(VI) only under anaerobic conditions at the expense of acetate as the electron donor . The bacterium was able to grow aerobically in L-broth supplemented with 15 mM CrO4(2-) but then did not reduce Cr(VI) . At a concentration of 2.4x10(9) cells/ml, 100 microM sodium chromate was reduced within 30 h; however, the maximum specific reduction rate was obtained at lower initial cell concentrations.

Cochrane Database Syst Rev . 2001;(3):CD002232.
Antibiotics for spontaneous bacterial peritonitis in cirrhotics; Soares-Weiser K et al.; BACKGROUND: Spontaneous bacterial peritonitis is mainly a complication of cirrhotic ascites that occurs in the absence of any intra-abdominal, surgically treatable source of infection . Antibiotics have been recommended as the mainstay treatment for spontaneous bacterial peritonitis . However, this recommendation is not based on convincing evidence . It has been proposed that treatment should cover Gram-negative enteric bacteria and Gram-positive cocci, that are responsible for up to 90% of cases . OBJECTIVES: To evaluate the effectiveness and safety of different types and ways of antibiotic therapy for spontaneous bacterial peritonitis in cirrhotic patients . SEARCH STRATEGY: Electronic searches on the Cochrane Library (Issue 3, 2000), the Cochrane Hepato-Biliary Group Trials Register (March 2000), EMBASE (1980-2000), MEDLINE (1966-2000); scanning the references of all identified studies; contacting the first author of each included trial . SELECTION CRITERIA: Randomised trials comparing different types of antibiotics for spontaneous bacterial peritonitis in cirrhotic patients . DATA COLLECTION AND ANALYSIS: Data were independently extracted by two reviewers . Relative risks or weighted mean differences, with their 95% confidence intervals were estimated using 'intention-to-treat' analyses . MAIN RESULTS: Nine trials dealing with 684 patients diagnosed with spontaneous bacterial peritonitis were included . No placebo-controlled trial was found . Each of the included trials compared different antibiotics, and no meta-analysis could be performed . We were unable to establish the optimal dose or duration of antibiotic therapy and found no convincing evidence that cefotaxime is more effective than ampicillin-tobramycin or that oral quinolones should be recommended for patients with less severe manifestations of the disease . REVIEWER'S CONCLUSIONS: This review provides no clear evidence for the treatment of cirrhotic patients with spontaneous bacterial peritonitis . Until large, well-conducted, trials provide adequate evidence, treatment must be based on clinical experience.

Support Care Cancer, 2001 Oct, 9(7), 514 - 21
Infectious complications in children with acute lymphoblastic leukemia and T-cell lymphoma--a rationale for tailored supportive care; Lex C et al.; Infections still remain a major cause of therapy-associated morbidity and death in patients with malignant diseases . To further lower the risk of serious and long-lasting infections by additional supportive measures, detailed information on the frequency and characteristic features of infections is needed . Therefore, patient data from 112 children with acute lymphoblastic leukemia and T-cell lymphoma who were treated according to the COALL-05-92 protocol in our department were analyzed for differences in the frequency and origin of febrile episodes in relation to age, immunological type of leukemia, treatment in group assessed as being at high or low risk of relapse, actual occurrence of relapse, and course of chemotherapy . At the time of diagnosis, low-risk patients more commonly presented with febrile episodes than high-risk patients . In total, patients developed a fever in 313 (24%) of 1,307 evaluated chemotherapy cycles . Febrile episodes were associated with microbiologically or clinically documented infections in 60% of all cases, while in 40% the fever was of unknown origin . Gram-positive pathogens had a markedly higher incidence than gram-negative or fungal ones . The incidence of febrile episodes during therapy appeared to be correlated with certain chemotherapeutic drug combinations . The highest rate was found after high-dose cytarabine and asparaginase causing a long period of leukopenia . However, after other chemotherapy courses with a similar duration of leukopenia the incidence of febrile episodes was significantly lower, suggesting that specific interactions of different chemotherapeutic agents with the immune response might be an important factor in development of infections . Individual factors might also account for an increased incidence of infections, since the number of high-risk patients with recurrent infections was significantly higher than expected on the basis of statistical evaluation . In conclusion, our findings suggest that the risk of infections during chemotherapy may not only be influenced by leukopenia, but that drug-specific effects of the various chemotherapeutic agents and individual factors may also be important contributory factors . These observations must be further expanded in prospective studies so that new tailored supportive care protocols can be elaborated.

Int J Antimicrob Agents, 2001 Sep, 18(3), 239 - 43
Vancomycin levels in human aqueous humour after intravenous and subconjunctival administration; Souli M et al.; The purpose of the present study was to evaluate the level of vancomycin in human aqueous humour after intravenous (i.v.) and subconjunctival administration . One hundred patients scheduled to undergo cataract extraction participated in the study . Fifty-three of them received 20 mg vancomycin subconjunctivally and 47 received two doses of vancomycin i.v . (1 g b.i.d.) . Specimens of aqueous humour from the first group were collected 1, 2, 2.5, 3, 3.5, 5, 6, 7, and 8 h after the subconjunctival injection . In the second group, specimens of blood and aqueous humour were collected 1, 2, 4, 6, 8, 10, and 12 h after the end of infusion of the second dose of the antibiotic . The levels of vancomycin were determined by fluorescent polarization immunoassay . In the first group peak levels of 24.82+/-3.55 microg/ml were achieved in aqueous humour at 5 h, whereas in the second group peak levels of 1.42+/-0.47 microg/ml were detected at 6 h . The latter levels, although higher than the MICs of most of the Gram-positive pathogens causing eye infections, are inadequate for the treatment of infections in vivo . These results support the need for subconjunctival instead of i.v . administration of vancomycin in order to achieve therapeutic levels of the drug in human aqueous humour or for prophylactic use whenever indicated.

Presse Med, 2001 Sep 8, 30(25 Pt 2), XIX - XXII
{Value of Synercid in clinical practice: from temporary approval to clinical trial authorization}; Potel G; NOSOCOMIAL PNEUMONIA DUE TO GRAM-POSITIVE COCCI: In a randomized multicentric trial comparing Synercid with vancomycin, the cure rate (56.3% vs 58.3%) were equivalent in the 2 treatment arms . Treatment failures were also similar: 44% vs 42% . Mortality (25% vs 22%) was likewise comparable, as was tolerance . SKIN AND SOFT TISSUE INFECTIONS: For erysipela, infections requiring surgical dissection, post-trauma infections, postoperative wound infections, or diabetes-related infections, the rate of success obtained in 2 open randomized comparative multicentric trials was equivalent in the 2 treatment arms: 68.2% for Syncercid, 70.7% for the compared treatments . EMERGENCY PRESCRIPTION: For E . faecium, the success rate was 74% based on clinical assessment and 70.5% based on bacteriological assessment . For meti-S S . aureus infections, the clinical success rate was 74% for all patients and 80% for bacteriologically evaluable patients; the bacteriological success rate was 74% and 71% respectively . In case of infection due to C-MLSB meti-R S . aureus, the percentage of clinical success was 89% for bacteriologically evaluable patients.

Crit Rev Microbiol, 2001, 27(3), 201 - 22
Chromobacterium violaceum: a review of pharmacological and industiral perspectives; Duran N et al.; Violet-pigmented bacteria, which have been described since the end of the 19th century, are occasionally the causative agent of septicemia and sometimes cause fatal infection in human and animals . Bacteria, producing violet colonies due to the production of a nondiffusible pigment violacein, were classified as a redefined genus Chromobacterium . Chromobacterium violaceum is gram-negative, and saprophyte from soil and water is normally considered nonpathogenic to human, but is an opportunistic pathogen of extreme virulence for human and animals . The biosynthesis and biological activities of violacein and the diverse effects of this pigment have been studied . Besides violacein, C . violaceum produces other antibiotics, such as aerocyanidin and aerocavin, which exhibit in vitro activity against both gram-negative and gram-positive bacteria . 3,6-Dihydroxyindoxazene and Y-TO678h exhibit a selective activity against gram-negative bacteria . Arphamenine A and B, and FR901228, that enhanced immunoresponse, and potentiators of beta-lactam antibiotics and chelators such as ferrioxamine exhibit important clinical potential applications . Lipopolysaccharides and polyhydroxyesters together with several enzymes appear as important metabolites with biotechnological applications . Many of these metabolites were already studied at the genome level.

Am J Vet Res, 2001 Oct, 62(10), 1664 - 9
Effects of antibiotics on morphologic characteristics and migration of canine corneal epithelial cells in tissue culture; Hendrix DV et al.; OBJECTIVE: To determine effects of commonly used ophthalmic antibiotics on cellular morphologic characteristics and migration of canine corneal epithelium in cell culture . SAMPLE POPULATION: Corneal epithelial cells harvested from corneas of 12 euthanatized dogs and propagated in cell culture . PROCEDURE: Cells were treated with various antibiotics after a defect was created in the monolayer . Cellular morphologic characteristics and closure of the defect were compared between antibiotic-treated and control cells . RESULTS: Cells treated with ciprofloxacin and cefazolin had the greatest degree of rounding, shrinkage, and detachment from plates . Cells treated with neomycin-polymyxin B-gramicidin and gentamicin sulfate had rounding and shrinkage but with less detachment . Cells treated with tobramycin and chloramphenicol grew similarly to control cells . On the basis of comparisons of defect circumference between control cells and cells exposed to antibiotics, tobramycin affected cellular migration the least . CONCLUSIONS AND CLINICAL RELEVANCE: Effects of ciprofloxacin and cefazolin on morphologic characteristics of canine corneal epithelial cells in vitro should be taken into consideration before using these antibiotics for first-line of treatment for noninfected ulcers . Of the antibiotics tested that have a primarily gram-negative spectrum of coverage, gentamicin inhibited corneal epithelial cell migration and had greater cytopathologic effects than tobramycin did . For antibiotics with a gram-positive coverage, chloramphenicol had no cytopathologic effects on cells in comparison to cefazolin, which caused most of the cells to shrink and detach from the plate . Polymyxin B-neomycin-gramicidin was midrange in its effects on cellular morphologic characteristics and migration.

Curr Opin Microbiol, 2001 Oct, 4(5), 509 - 14
Efflux-mediated drug resistance in Gram-positive bacteria; Markham PN et al.; Gram-positive bacteria express numerous membrane transporters that promote the efflux of various drugs, including many antibiotics, from the cell to the outer medium . Drug transporters can be specific to a particular drug, or can have broad specificity, as in so-called multidrug transporters . This broad specificity can be a consequence of the hydrophobic nature of transported molecules, as suggested by recent structural studies of soluble multidrug-binding proteins . Although the functions of drug transporters may involve both the protection of bacteria from outside toxins and the transport of natural metabolites, their clinical importance lies largely in providing Gram-positive pathogens with resistance to macrolides, tetracyclines and fluoroquinolones . A number of agents, discovered in recent years, that inhibit drug transporters can potentially be used to overcome efflux-associated antibiotic resistance.

Perit Dial Int, 2001 Jul-Aug, 21(4), 360 - 4
Diverticular disease and treatment with gastric acid inhibitors do not predispose to peritonitis of enteric origin in peritoneal dialysis patients; del Peso G et al.; OBJECTIVE: Enteric peritonitis (EP) is an infrequent complication of peritoneal dialysis (PD), with severe consequences for peritoneal membrane viability and patient outcome . Factors such as diverticular disease and gastric acid inhibitors have been implicated in its appearance . We investigated several risk factors, including those mentioned below, that can influence the development of EP . DESIGN: Retrospective cross-sectional study . SETTING: Tertiary-care public university hospital . PATIENTS: Fifty-seven PD patients treated in our PD unit during August 1998 . MAIN OUTCOME MEASURES: A barium enema was performed on 50 of the 57 patients (the remaining 7 patients refused it) in order to exclude the presence of diverticulosis . All episodes of peritonitis occurring in those patients, including EP, were registered . Enteric peritonitis was defined as that caused by gram-positive, gram-negative, or fungus micro-organisms that colonized the intestinal tract, excluding episodes secondary to genitourinary tract or peritoneal catheter exit-site infections . RESULTS: Twenty-four patients showed diverticular disease in the barium enema, but only 5 of them (21%) had any EP episode . Five of the 26 patients with no diverticula (19%) had EP . Fifty-five episodes of peritonitis were reported in 21 patients; 15 episodes of EP (27.3% of all) developed in 11 patients . Seven of the 11 patients (64%) required peritoneal catheter removal and 3 of them (27%) finally were transferred to hemodialysis due to consequences of the EP episode . Logistic regression analysis did not find any of the independent variables analyzed (age, sex, time on PD, type of PD, peritoneal transport parameters, presence of polycystic kidney disease, constipation or diverticulosis, or treatment with gastric acid inhibitors, or phosphate-binding agents) to be risk factors for developing EP . CONCLUSIONS: Neither diverticulosis nor treatment with gastric acid inhibitors seem to be risk factors for developing peritonitis of enteric origin in PD patients . This type of peritonitis has to be promptly identified and treated in order to diminish the high frequency of peritoneal catheter removal and PD dropout due to such episodes.

J Biol Chem, 2001 Dec 7, 276(49), 45840 - 7 Epub 2001 Oct 03.
Distinct carbohydrate recognition domains of an invertebrate defense molecule recognize Gram-negative and Gram-positive bacteria; Bilej M et al.; Coelomic fluid of Eisenia foetida earthworms (Oligochaeta, Annelida) contains a 42-kDa defense molecule named CCF for coelomic cytolytic factor . By binding microbial antigens, namely the O-antigen of lipopolysaccharide (LPS), beta-1,3-glucans, or N,N'-diacetylchitobiose present, respectively, on Gram-negative bacteria or yeast cell walls, CCF triggers the prophenoloxidase activating pathway . We report that CCF recognizes lysozyme-predigested Gram-positive bacteria or the peptidoglycan constituent muramyl dipeptide as well as muramic acid . To identify the pattern recognition domains of CCF, deletion mutants were tested for their ability to reconstitute the prophenoloxidase cascade in E . foetida coelomic fluid depleted of endogenous CCF in the presence of LPS, beta-1,3-glucans, N,N'-diacetylchitobiose, and muramic acid . In addition, affinity chromatography of CCF peptides was performed on immobilized beta-1,3-glucans or N,N'-diacetylchitobiose . We found that the broad specificity of CCF for pathogen-associated molecular patterns results from the presence of two distinct pattern recognition domains . One domain, which shows homology with the polysaccharide and glucanase motifs of beta-1,3-glucanases and invertebrate defense molecules located in the central part of the CCF polypeptide chain, interacts with LPS and beta-1,3-glucans . The C-terminal tryptophan-rich domain mediates interactions of CCF with N,N'-diacetylchitobiose and muramic acid . These data provide evidence for the presence of spatially distinct carbohydrate recognition domains within this invertebrate defense molecule.

J Hepatol, 2001 Aug, 35(2), 156 - 63
Scavenger cells with gram-positive bacterial lipoteichoic acid infiltrate around the damaged interlobular bile ducts of primary biliary cirrhosis; Tsuneyama K et al.; BACKGROUND/AIMS: Gram-positive bacterial DNA is frequently detectable in gallbladder bile of primary biliary cirrhosis (PBC) patients . To advance these findings, lipoteichoic acid (LTA) of gram-positive bacteria with high antigenicity was examined in liver specimens and bile from PBC patients and controls . METHODS: LTA was examined by Western blotting in the gallbladder bile from 15 PBC, 11 cholecystolithiasis and six normal subjects, and by immunohistochemistry in liver specimens from 16 PBC, six primary sclerosing cholangitis (PSC), eight chronic viral hepatitis C (CVH-C) and five normal subjects . RESULTS: In the gallbladder bile, there was no significant difference in the positive rate of LTA between PBC and controls . LTA-containing mononuclear cells were frequently detected in the portal tracts, particularly around the bile ducts and in hepatic sinusoids in PBC, while they were infrequent or occasional in control livers . These LTA-containing cells were sinusoidal endothelial cells and Kupffer cells, and portal monocytes, which frequently expressed scavenger receptor class B type 1 . CONCLUSIONS: LTA derived from bacterial fragments may reach the bile, not only in the diseased state but also under normal conditions . Such LTA may be involved in the development and progression of portal tract lesions, particularly bile duct lesions, in PBC.

Nihon Kokyuki Gakkai Zasshi, 2001 Jul, 39(7), 492 - 7
{Pneumonia caused by Nocardia nova}; Ikeue T et al.; This is the first clinical report of a case of pneumonia caused by Nocardia nova in Japan . A 52 year-old woman who had received steroids and cyclophosphamide for six years because of polymyositis was admitted to our hospital for further examination . On admission she had a mild cough, and her chest radiography and computed tomography revealed bilateral multiple nodules, some of which were cavitated . She developed a cough productive of yellow sputum and fever up to 38 degrees C . Examination of the sputum revealed a gram-positive branched organism and sputum cultures repeatedly grew Nocardia species . The isolate was identified as Nocardia nova later . Clinical recovery was obtained readily upon treatment with imipenem and trimethoprim methoxazole, though the latter drug was discontinued because of nausea and anorexia . This drug was therefore replaced with oral minocycline, which proved to be ineffective clinically although susceptibility testing of the drug showed positive sensitivity . Minocycline was replaced with clarithromycin, after which chest radiography and computed tomography showed almost total resolution of the infiltrates . Clarithromycin may be an alternative oral agent to sulfonamides or minocycline when these agents are ineffective or not tolerated.

Parasitology, 2001 Sep, 123(Pt 3), 271 - 6
The interaction between the gelatin-binding domain of fibronectin and the attachment of Pasteuria penetrans endospores to nematode cuticle; Mohan S et al.; Pasteuria penetrans is a Gram-positive endospore-producing bacterium that is a parasite of root-knot nematodes . Attachment of endospores to the cuticle of the nematode is the first stage in the infection process . Western blot analysis with monoclonal and polyclonal antibodies that recognize the 30 kDa heparin-binding domain (HBD) and the 45 kDa gelatin-binding domain (GBD) fragments of human fibronectin (Fn) revealed a series of polypeptides of approximately 40, 45 and 55 kDa present in crude cuticle extracts of Meloidogyne javanica 2nd-stage juveniles . The results suggest that the structure of the nematode fibronectin is different to the fibronectins so far characterized . Pre-treatment of endospores of Pasteuria with either the HBD or the GBD was found to inhibit binding to the nematode cuticle . The larger GBD fragment was the most effective at blocking adhesion . Pre-treatment of the GBD fragment with gelatin prevented the GBD fragment from inhibiting endospore attachment to the nematode cuticle.

Biosci Biotechnol Biochem, 2001 Aug, 65(8), 1897 - 9
Enzymatic glycosylation of lincomycin; Weignerova L et al.; Lincomycin (1), a glycosidic antibiotic, active against Gram-positive bacteria, was modified enzymatically with the aim of improving its physico-chemical and biological properties . Compound 1 was glycosylated using jack bean alpha-mannosidase to produce 7-O-alpha-D-mannopyranosyl-lincomycin (2).

Biosci Biotechnol Biochem, 2001 Aug, 65(8), 1774 - 81
Biotransformation of phenanthrene and 1-methoxynaphthalene with Streptomyces lividans cells expressing a marine bacterial phenanthrene dioxygenase gene cluster; Chun HK et al.; The phdABCD gene cluster in a marine bacterium Nocardioides sp . strain KP7 codes for the multicomponent enzyme phenanthrene dioxygenase . phdA encoding an iron-sulfur protein large subunit alpha, phdB encoding its small subunit beta, phdC encoding ferredoxin, and phdD encoding ferredoxin reductase, were replaced in such a way that the termination codons of the preceding open reading frames were overlapped with the initiation codons of the following genes . This manipulated phdABCD gene cluster was positioned downstream of the thiostrepton-inducible promoter PtipA in a high-copy-number vector pIJ6021, and introduced into the gram-positive, soil-inhabiting, filamentous bacterium Streptomyces lividans . The recombinant S . lividans cells converted phenanthrene into a cis-diol form, which was determined to be cis-3,4-dihydroxy-3,4-dihydrophenanthrene by its UV spectral data as well as HPLC property, using the authentic sample for comparison . This biotransformation proceeded very efficiently; 200 microM and 2 mm of phenanthrene were almost completely converted to its cis-diol form in 6 h and 32 h, respectively . In addition, the S . lividans cells carrying the phdABCD gene cluster were found to transform 1-methoxynaphthalene to two products, which were identified to be 8-methoxy-2-naphthol in addition to 8-methoxy-1,2-dihydro-1,2-naphthalenediol by their EI-MS, 1H- and 13C-NMR spectral data.

Int J Antimicrob Agents, 2001, 18 Suppl 1, S57 - 61
Community-acquired pneumonia in ambulatory patients: relative importance of atypical pathogens; Honeybourne D; The presence of atypical pathogens in community-acquired pneumonia is difficult to diagnose; culture is time-consuming and requires considerable expertise and serological identification may be inaccurate . Nevertheless, the increasing importance of atypical organisms has been recognised in recent years . Variations in aetiology have been detected in different geographical regions and different patient populations . The season of the year may also influence aetiology, and some infections follow a cyclical pattern . When an atypical pathogen is suspected, a macrolide antibiotic is an appropriate choice as, in addition to activity against these organisms, they are usually effective against the Gram-positive and Gram-negative respiratory pathogens implicated in community-acquired pneumonia.

Int J Antimicrob Agents, 2001, 18 Suppl 1, S25 - 8
Macrolide resistance mechanisms in Gram-positive cocci; Pechere JC; Two principal mechanisms of resistance to macrolides have been identified in Gram-positive bacteria . Erythromycin-resistant methylase is encoded by erm genes . Resultant structural changes to rRNA prevent macrolide binding and allow synthesis of bacterial proteins to continue . Presence of the erm gene results in high-level resistance . Modification of the mechanism whereby antibiotics are eliminated from the bacteria also brings about resistance . Bacteria carrying the gene encoding macrolide efflux (i.e . the mefE gene) display relatively low-level resistance . Azithromycin, because of its ability to achieve concentrations at sites of infections, is capable of eradicating mefE-carrying strains . Other resistance mechanisms, involving stimulation of enzymatic degradation, appear not to be clinically significant.

Curr Opin Crit Care, 2001 Aug, 7(4), 232 - 7
New anti-Gram-positive agents; Hamilton DC et al.; As the prevalence of resistant Gram-positive organisms in the critical care unit has increased, so have the associated morbidity and mortality and the cost of their treatment . As a result, more toxic and less active second-line agents and combinations of agents are used, often with limited evidence of clinical benefit . Although widely used, the role of glycopeptides is limited by increasing resistance and poor pharmacokinetics . New agents now in use, such as quinupristin/dalfopristin and Linezolid (Zyvox; Pharmacia & Upjohn, Kalamazoo, MI), show promise, as do diverse agents in development . The future is likely to bring greater therapeutic choice but, inevitably, further resistance.

Nat Prod Lett, 2001, 15(2), 93 - 101
Bioactive compounds from Iostephane heterophylla (Asteraceae); Aguilar MI et al.; The novel bisabolene sesquiterpenes 3-6, were isolated from Iostephane heterophylla, using bioguided fractionation . The new compounds were determined to be (12R/12S)-12,13-epoxy-xanthorrhizols (3,4) and (12R/12S)-12,13-dihydro-12,13-dihydroxy-xanthorrizols (5,6) and their structures were characterized by analysis of spectroscopic data and by chemical correlation from xanthorrhizol (2) . The stereochemistry at C-12 of 5 was deduced using the modified Mosher experiment . Some of the isolated compounds elicited activity against gram positive and gram negative bacteria, levadura and dermatophytes.

J Cell Sci, 2001 Jul, 114(Pt 13), 2535 - 45
Fluorescence recovery after photobleaching reveals that LPS rapidly transfers from CD14 to hsp70 and hsp90 on the cell membrane; Triantafilou K et al.; Although CD14 has been implicated in the immune recognition of bacterial lipopolysaccharide (LPS) from Gram-negative bacteria and also peptidoglycan (PGN) and lipoteichoic acid (LTA) from the outer cell wall of Gram-positive bacteria, accumulating evidence has suggested the possible existence of other functional receptor(s) . In this study, we have used fluorescence recovery after photobleaching (FRAP) in order to get the first dynamic picture of the innate recognition of bacteria . We have found that the diffusion coefficient of CD14 remains unaffected after LPS ligation and that the diffusion coefficients of FITC-LPS and FITC-LTA bound to cells differ from that of CD14 . Furthermore, FITC-LPS/LTA rapidly become immobile when bound to cells, suggesting that FITC-LPS/LTA must briefly associate with CD14 in the initial attachment process and rapidly move on to an immobile receptor or to a complex of receptors . Further FRAP experiments revealed that heat shock protein 70 (hsp70) and hsp90 are immobile in cell membranes, and antibodies against them were found to block the transfer of LPS to the immobile receptor and to inhibit interleukin 6 production upon LPS stimulation . These experiments indicated that LPS transfers from CD14 to hsp70 and hsp90, which may be part of an LPS/LTA multimeric receptor complex . Thus, hsps are implicated as mediators of the innate activation by bacteria.

Chem Res Toxicol, 2001 Sep, 14(9), 1259 - 65
Molecular requirements for inhibition of cytochrome p450 activities by roquefortine; Aninat C et al.; Roquefortine, a cyclopeptide derived from the diketopiperazine cyclo(Trp-dehydroHis), is a secondary metabolite produced by several Penicillium species . It has been reported to cause neurotoxic effect and to inhibit Gram-positive bacteria growth . The mechanisms responsible for its toxicity and metabolism are still unknown . In this study, we investigated the interaction of roquefortine with mammalian cytochromes P450 . Roquefortine interaction with rat and human liver cytochromes P450 was monitored by difference UV-vis spectroscopy . It was found to interact with different forms of the cytochromes, giving rise to a type II difference spectrum, characteristic of the binding of an amino function to the heme iron . Roquefortine exhibited high affinity for microsomes from rats treated with various inducers, the K(s) values being in the range 0.2-8 microM . Similar results were observed with human P450 enzymes 1A1, 1A2, 2D6, and 3A4 . Roquefortine had no effect on NAPDH cytochrome c reductase . Therefore, inhibition of NADPH consumption was observed using various rat liver microsomes alone or in the presence of 100 microM testosterone in the case of dexamethasone (DEX)-rat microsomes . Enzymatic inhibition was studied in terms of P450 3A activities, i.e., testosterone 6beta-hydroxylase (IC(50) around 10 microM) or bromocriptine metabolism (IC(50) > 50 microM) using DEX-rat liver microsomes or P450 3A4, benzphetamine N-demethylase using phenobarbital-rat liver microsomes (IC(50) > 30 microM), and ethoxyresorufin metabolism using 3-methylcholanthrene-rat liver microsomes (IC(50) 0.1 microM), P450 1A1, and 1A2 . Roquefortine was compared with compounds of similar structure: cyclo(Phe-His), cyclo(Phe-dehydroHis), cyclo(Trp-His), phenylahistin . These studies indicate that the =N- imidazole moiety coordinates with the heme iron, and suggest that the dehydroHis moiety and the presence of a fused tetracycle play an important part in roquefortine inhibitory power.

Kekkaku, 2001 Aug, 76(8), 593 - 600
{Bacterial infections and toll-like receptors}; Akira S; Toll-like receptors are type-1 transmembrane receptors involved in microbial recognition . TLR4 has been shown to function as the lipopolysaccharide signaling receptor, while TLR2 recognizes peptidoglycans from Gram-positive bacteria, and lipoproteins . TLR9 is involved in the recognition of bacterial DNA (CpG DNA) . Although various microbial cell wall components are recognized by different receptors, all of these responses are abrogated in MyD88-deficient cells . These results show that different TLRs recognize different microbial cell wall components, and that MyD88 is an essential signaling molecule shared among interleukin-1 receptor/Toll family members . However, in LPS signaling MyD88-independent pathway is present in addition to MyD88-dependent pathway.

Appl Microbiol Biotechnol, 2001 Aug, 56(3-4), 502 - 3
An antibiotic with activity against gram-positive bacteria from the gentamicin-producing strain of Micromonospora purpurea; Rusnak K et al.; A mixture of polycyclic aromatic compounds with activity against gram-positive bacteria was isolated from a gentamicin-producing species of Micromonospora.

Can J Microbiol, 2001 Jul, 47(7), 608 - 17
Analysis of pFQ12, a 22.4-kb Frankia plasmid; John TR et al.; Frankia are gram-positive, filamentous bacteria capable of fixing atmospheric dinitrogen in symbiosis with a wide variety of woody plants and shrubs . Some isolates of Frankia harbor plasmids of 8.5 (pFQ11) and 22.4 kb (pFQ12) that have no known function but are transmitted through many generations in culture . We have sequenced the 22,437-bp pFQ12 plasmid that is present in isolates CpI1 and ArI3 . This sequence, with 76% G+C, is almost totally unrelated to that of pFQ11 found in the same cells . However, four regions of identity, 40-90 bp each, are dispersed around the plasmids . The 22.4-kb plasmid has >50 open reading frames (ORFs) that encode putative proteins of more than 100 amino acids, with the largest being 2226 amino acids . Twenty of these ORFs are likely to encode proteins based on their codon bias as determined by two different algorithms . Transcripts from nine of these regions have been identified by reverse transcriptase-polymerase chain reaction (RT-PCR) or filter hybridization . The two Frankia plasmids each encode a protein similar to the korSA protein that regulates transmission of pSAM2 in Streptomyces . The origin of replication (ORI) region of pFQ12 was localized by intrastrand AT and GC equivalence switch . It includes a 40-bp, intergenic, A+T-rich region that has a strong identity in pFQ11.

Crit Care Med, 2001 Sep, 29(9), 1750 - 5
Nitric oxide-dependent down-regulation of angiotensin II type 2 receptors during experimental sepsis; Bucher M et al.; OBJECTIVE: The systemic renin-angiotensin system is highly activated during septic shock . This has focused interest in regulation of the adrenal angiotensin II type 2 receptor (AT2) as the target thought to mediate angiotensin II-induced adrenal catecholamine release during experimental sepsis in vivo . In addition, the influence of typical endogenous mediators of sepsis, such as proinflammatory cytokines and nitric oxide, on AT2 receptor expression should be investigated in vitro . DESIGN: Prospective animal trial followed by a controlled cell culture study . SETTING: Laboratory of the Department of Anesthesiology . SUBJECTS: Male Sprague-Dawley rats weighing 200-250 g, PC12 cell line . INTERVENTIONS: Rats were injected with lipopolysaccharide to stimulate Gram-negative sepsis or lipoteichoic acid to stimulate Gram-positive sepsis . AT2 receptor expression, abundance of the proinflammatory cytokines (interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma), and nitric oxide synthase II expression have been determined in the adrenal gland . Rat adrenal pheochromocytoma cells were incubated with these cytokines or with the nitric oxide donors sodium nitroprusside or S-nitroso-N-acetylpenicillamine to investigate the regulation of AT2 receptors during severe inflammation on a cellular level . MEASUREMENTS AND MAIN RESULTS: In the adrenal gland, AT2 receptor expression was down-regulated in both models of sepsis, whereas tissue cytokine concentrations were elevated and nitric oxide synthase II expression was induced . Incubation of PC12 cells with proinflammatory cytokines resulted in a dose-dependent diminished expression of AT2 receptors, which was mimicked by incubation with nitric oxide donors . Blocking of cytokine-induced nitric oxide synthesis by co-incubation of PC12 cells with NG-nitro-l-arginine methyl ester prevented down-regulation of AT2 receptors . CONCLUSIONS: These findings show that in our model of sepsis, the expression of AT2 receptors in the adrenal gland is down-regulated in a nitric oxide-dependent manner . Because AT2 receptors are thought to be involved in adrenal catecholamine secretion in a stimulatory fashion, the diminished expression of AT2 receptors could play an important role in the pathogenesis of septic shock via impaired angiotensin II-induced adrenal catecholamine release, despite a strong activation of the systemic renin-angiotensin system.

Adv Space Res, 1983, 3(8), 43 - 7
Characterization of a halotolerant-psychroloterant bacterium from dry valley Antarctic soil; Miller KJ et al.; The saline soils of the ice free dry valleys of Victoria Land, Antarctica may provide the closest analog on Earth to Martian conditions . We have initiated a study aimed at examining microbial adaptations to the harsh environment of these dry valley soils . In this report we describe the characterization of one bacterium, strain A4a, isolated from Taylor Valley soil . Strain A4a was an obligately aerobic, orange-pigmented, Gram-positive coccus that grew over wide ranges of both temperature (0 degrees C-40 degrees C) and sodium chloride concentration (0-2.0M) . The optimal temperature for growth at all NaCl concentrations was 25 degrees C . Phospholipid composition and guanine plus cytosine content of the DNA of the isolate indicate a close relation to the genus Planococcus.

J Microbiol Methods, 1984, 2, 165 - 76
Determination of the gram-positive bacterial content of soils and sediments by analysis of teichoic acid components; Gehron MJ et al.; Many gram-positive bacteria form substituted polymers of glycerol and ribitol phosphate esters known as teichoic acids . Utilizing the relative specificity of cold concentrated hydrofluoric acid in the hydrolysis of polyphosphate esters it proved possible to quantitatively assay the teichoic acid-derived glycerol and ribitol from gram-positive bacteria added to various soils and sediments . The lipids are first removed from the soils or sediments with a one phase chloroform-methanol extraction and the lipid extracted residue is hydrolyzed with cold concentrated hydrofluoric acid . To achieve maximum recovery of the teichoic acid ribitol, a second acid hydrolysis of the aqueous extract is required . The glycerol and ribitol are then acetylated after neutralization and analyzed by capillary gas-liquid chromatography . This technique together with measures of the total phospholipid, the phospholipid fatty acid, the muramic acid and the hydroxy fatty acids of the lipopolysaccharide lipid A of the gram-negative bacteria makes it possible to describe the community structure environmental samples . The proportion of gram-positive bacteria measured as the teichoic acid glycerol and ribitol is higher in soils than in sediments and increases with depth in both.

J Microbiol Biotechnol, 1997 Apr, 7(2), 95 - 100
Lysine epsilon-aminotransferase, the initial enzyme of cephalosporin biosynthesis in actinomycetes; Rius N et al.; Streptomyces clavuligerus, Streptomyces lipmanii and Nocardia (formerly Streptomyces) lactamdurans are Gram-positive mycelial bacteria that produce medically important beta-lactam antibiotics (penicillins and cephalosporins including cephamycins) that are synthesized through a series of reactions starting from lysine, cysteine and valine . L-lysine epsilon-aminotransferase (LAT) is the initial enzyme in the two-step conversion of L-lysine to L-alpha-aminoadipic acid, a specific precursor of all penicillins and cephalosporins . Whereas S . clavuligerus uses LAT for cephalosporin production, it uses the cadaverine pathway for catabolism when lysine is the nitrogen source for growth . Although the cadaverine path is present in all examined streptomycetes, the LAT pathway appears to exist only in beta-lactam-producing strains . Genetically increasing the level of LAT enhances the production of cephamycin . LAT is the key rate-limiting enzyme in cephalosporin biosynthesis in S . clavuligerus strain NRRL 3585 . This review will summarize information on this important enzyme.

Photosynth Res, 1994, 41, 285 - 94
Evolution of heliobacteria: implications for photosynthetic reaction center complexes; Vermaas WF; The evolutionary position of the heliobacteria, a group of green photosynthetic bacteria with a photosynthetic apparatus functionally resembling Photosystem I of plants and cyanobacteria, has been investigated with respect to the evolutionary relationship to Gram-positive bacteria and cyanobacteria . On the basis of 16S rRNA sequence analysis, the heliobacteria appear to be most closely related to Gram-positive bacteria, but also an evolutionary link to cyanobacteria is evident . Interestingly, a 46-residue domain including the putative sixth membrane-spanning region of the heliobacterial reaction center protein show rather strong similarity (33% identity and 72% similarity) to a region including the sixth membrane-spanning region of the CP47 protein, a chlorophyll-binding core antenna polypeptide of Photosystem II . The N-terminal half of the heliobacterial reaction center polypeptide shows a moderate sequence similarity (22% identity over 232 residues) with the CP47 protein, which is significantly more than the similarity with the Photosystem I core polypeptides in this region . An evolutionary model for photosynthetic reaction center complexes is discussed, in which an ancestral homodimeric reaction center protein (possibly resembling the heliobacterial reaction center protein) with 11 membrane-spanning regions per polypeptide has diverged to give rise to the core of Photosystem I, Photosystem II, and of the photosynthetic apparatus in green, purple, and heliobacteria.

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