FEBS J, 2005 Jan, 272(2), 375 - 89 Characterization of the secreted chorismate mutase from the pathogen Mycobacterium tuberculosis; Sasso S et al.; The gene encompassing ORF Rv1885c with weak sequence similarity to AroQ chorismate mutases (CMs) was cloned from the genome of Mycobacterium tuberculosis and expressed in Escherichia coli . The gene product (*MtCM) complements a CM-deficient E . coli strain, but only if produced without the predicted N-terminal signal sequence typical of M . tuberculosis . The mature *MtCM, which was purified by exploiting its resistance to irreversible thermal denaturation, possesses high CM activity in vitro . The enzyme follows simple Michaelis-Menten kinetics, having a k(cat) of 50 s(-1) and a K(m) of 180 microm (at 30 degrees C and pH 7.5) . *MtCM was shown to be a dimer by analytical ultracentrifugation and size-exclusion chromatography . Secondary-structure prediction and CD spectroscopy confirmed that *MtCM is a member of the all-alpha-helical AroQ class of CMs, but it seems to have a topologically rearranged AroQ fold . Because CMs are normally intracellular metabolic enzymes required for the biosynthesis of phenylalanine and tyrosine, the existence of an exported CM in Gram-positive M . tuberculosis is puzzling . The observation that homologs of *MtCM with a predicted export sequence are generally only present in parasitic or pathogenic organisms suggests that secreted CMs may have evolved to participate in some aspect of parasitism or pathogenesis yet to be unraveled. Clin Exp Immunol, 2005 Feb, 139(2), 328 - 37 Identification of epithelial auto-antigens associated with periodontal disease; Ye P et al.; Summary We previously reported evidence that patients with periodontitis have serum antibodies to oral Gram positive bacteria that are cross-reactive with epithelial antigens . In the present report cross-reactive epithelial antigens including CD24, lactate dehydrogenase A {LDM-A}, antioxidant protein 2 {AOP 2} and nuclear factor of activated T cells 5 {NFAT 5}, were identified by screening a cDNA expression library with pooled patient sera . Titres of antibodies to CD24 peptide correlated negatively with indices of periodontal disease severity . Strong expression of CD24 in the reactive periodontal epithelium and inflamed gingival attachment contrasted with low to undetectable expression in the external gingival epithelium . In periodontitis, a local action of these auto-reactive antibodies could modulate the regulatory potential associated with expression of CD24 in this epithelium. Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 457 - 62 Arthrobacter bergerei sp . nov . and Arthrobacter arilaitensis sp . nov., novel coryneform species isolated from the surfaces of cheeses; Irlinger F et al.; Fourteen isolates of two different bacterial species isolated from the surface of smear-ripened cheeses were found to exhibit many characteristics of the genus Arthrobacter . The isolates were aerobic, Gram-positive, catalase-positive, non-spore-forming and non-motile . The cell-wall peptidoglycan contained lysine, alanine and glutamic acid . rrs sequence analysis indicated that the new isolates Re117(T) and Ca106(T) are closely related to the Arthrobacter nicotianae group and showed highest sequence similarity (>98 %) to Arthrobacter nicotianae and Arthrobacter protophormiae . However, DNA-DNA hybridization studies indicated that the strains represented two novel genomic species within the genus Arthrobacter and did not belong to A . nicotianae or A . protophormiae (<43 % DNA-DNA relatedness) . On the basis of the phylogenetic and phenotypic distinctiveness of the new isolates, these bacteria should be classified as two novel Arthrobacter species, for which the names Arthrobacter bergerei sp . nov . and Arthrobacter arilaitensis sp . nov . are proposed . Type strains have been deposited in culture collections as Arthrobacter bergerei Ca106(T) (=CIP 108036(T)=DSM 16367(T)) and Arthrobacter arilaitensis Re117(T) (=CIP 108037(T)=DSM 16368(T)). Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 427 - 31 Actinomyces dentalis sp . nov., from a human dental abscess; Hall V et al.; A previously undescribed filamentous, beaded, Gram-positive, rod-shaped bacterium was isolated from pus of a human dental abscess . Based on its cellular morphology and the results of biochemical testing the organism was tentatively identified as a member of the genus Actinomyces, but it did not correspond to any currently recognized species of this genus . Comparative 16S rRNA gene sequencing studies showed the bacterium represents a distinct subline within the genus Actinomyces, clustering within a group of species that includes Actinomyces bovis, the type species of the genus . Sequence divergence values of >8 % with other recognized species within this phylogenetic group clearly demonstrated that the organism represents a hitherto unknown species . Based on biochemical and molecular phylogenetic evidence, it is proposed that the unidentified organism recovered from a dental abscess be classified as a novel species, Actinomyces dentalis sp . nov . The type strain is R18165(T) (=CCUG 48064(T)=CIP 108337(T)). Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 345 - 51 Marinilactibacillus piezotolerans sp . nov., a novel marine lactic acid bacterium isolated from deep sub-seafloor sediment of the Nankai Trough; Toffin L et al.; A piezotolerant, mesophilic, marine lactic acid bacterium (strain LT20(T)) was isolated from a deep sub-seafloor sediment core collected at Nankai Trough, off the coast of Japan . Cells were Gram-positive, rod-shaped, non-sporulating and non-motile . The NaCl concentration range for growth was 0-120 g l(-1), with the optimum at 10-20 g l(-1) . The temperature range for growth at pH 7.0 was 4-50 degrees C, with the optimum at 37-40 degrees C . The optimum pH for growth was 7.0-8.0 . The optimum pressure for growth was 0.1 MPa with tolerance up to 30 MPa . The main cellular phospholipids were phosphatidylglycerols (25 %), diphosphatidylglycerols (34 %) and a group of compounds tentatively identified as ammonium-containing phosphatidylserines (32 %); phosphatidylethanolamines (9 %) were minor components . The fatty acid composition was dominated by side chains of 16 : 0, 14 : 0 and 16 : 1 . The G+C content of the genomic DNA was 42 mol% . On the basis of 16S rRNA gene sequence analysis and the secondary structure of the V6 region, this organism was found to belong to the genus Marinilactibacillus and was closely related to Marinilactibacillus psychrotolerans M13-2(T) (99 %), Marinilactibacillus sp . strain MJYP.25.24 (99 %) and Alkalibacterium olivapovliticus strain ww2-SN4C (97 %) . Despite the high similarity between their 16S rRNA gene sequences (99 %), the DNA-DNA hybridization levels were less than 20 % . On the basis of physiological and genetic characteristics, it is proposed that this organism be classified as a novel species, Marinilactibacillus piezotolerans sp . nov . The type strain is LT20(T) (=DSM 16108(T)=JCM 12337(T)). Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 153 - 7 Agromyces salentinus sp . nov . and Agromyces neolithicus sp . nov; Jurado V et al.; A polyphasic study was carried out to clarify the taxonomic position of two Gram-positive bacteria isolated from soil samples of the Grotta dei Cervi (Italy), a relatively unexplored hypogean environment . The strains, 20-5(T) and 23-23(T), showed phenotypic and phylogenetic characteristics that were consistent with their classification in the genus Agromyces . 16S rRNA gene sequence comparisons revealed that the two strains formed distinct phyletic lines within the genus Agromyces . Based on 16S rRNA gene sequence similarity, chemotaxonomic data and the results of DNA-DNA relatedness studies, it is proposed that the two isolates represent two novel species of the genus Agromyces . Pronounced differences in a broad range of phenotypic characteristics and DNA G+C content distinguished the two strains from each other and from previously described species of the genus Agromyces . Two novel species are proposed: Agromyces salentinus sp . nov . (type strain, 20-5(T)=HKI 0320(T)=DSM 16198(T)=NCIMB 13990(T)) and Agromyces neolithicus sp . nov . (type strain, 23-23(T)=HKI 0321(T)=DSM 16197(T)=NCIMB 13989(T)). Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 139 - 42 Reclassification of strain CCM 132, previously classified as Kocuria varians, as Kocuria carniphila sp . nov; Tvrzova L et al.; A Gram-positive actinobacterium, previously classified as Kocuria varians, was subjected to a polyphasic taxonomic study . The bacterium showed the peptidoglycan type Lys-Ala(3) (variation A3alpha), MK-7(H(2)) was the major menaquinone and anteiso-C(15 : 0) and anteiso-C(17 : 0) were the major fatty acids . On the basis of the phylogenetic and phenotypic characteristics of the actinobacterium, a novel species, Kocuria carniphila sp . nov . (type strain, CCM 132(T)=DSM 16004(T)), is proposed. Int J Syst Evol Microbiol, 2005 Jan, 55(Pt 1), 67 - 70 Microbacterium halotolerans sp . nov., isolated from a saline soil in the west of China; Li WJ et al.; A Gram-positive short rod isolated from a saline soil in China was characterized using a polyphasic approach . This actinobacterium grew over a wide salinity range {0-15 % NaCl, 0-20 % KCl and 0-30 % MgCl(2).6H(2)O (w/v); optimum concentrations for growth were 5 % NaCl, 5-10 % KCl, 10 % MgCl(2).6H(2)O} . The optimum growth temperature and pH were 28-30 degrees C and 7.0-8.0, respectively . Chemotaxonomic features (peptidoglycan-type B2beta with glycolyl residues; major menaquinones MK-10 and MK-11; predominating iso- and anteiso-branched cellular fatty acids; DNA G+C content 66.5 mol%) placed this organism within the genus Microbacterium . 16S rRNA gene sequence analysis confirmed this classification of the strain, but showed that it was distinct from its nearest neighbours . It formed a separate branch with type strains of the genus Microbacterium, and also shared low sequence similarity with them (<96 %) . Based on phenotypic and molecular taxonomic results, it is proposed that the unknown isolate should be classified as a novel species in the genus Microbacterium, for which the name Microbacterium halotolerans sp . nov . is proposed . The type strain is YIM 70130(T) (=KCTC 19017(T)=CIP 108071(T)). Genetika, 2004 Dec, 40(12), 1717 - 21 {Distribution of transposons Tn5044 and Tn5070 with unusual mer operons in environmental bacterial populations}; Oxygen et al.; In this issue of Structure, report the structure of the redox-sensing repressor from the gram-positive bacterium Thermus aquaticus (T-Rex), a protein that links gene expression to oxygen limitation and the metabolic state of the cell. Biomacromolecules, 2005 Jan 10, 6(1), 180 - 188 Identification and Characterization of Genes from Streptomycessp . Strain K30 Responsible for Clear Zone Formation on Natural Rubber Latex and Poly(cis-1,4-isoprene) Rubber Degradation; Rose K et al.; Streptomyces sp . strain K30 was isolated from soil next to a city high way in Munster (Germany) according to its ability to degrade natural and synthetic poly(cis-1,4-isoprene) rubber and to form clear zones on natural rubber latex agar plates . The clear zone forming phenotype was used to clone the responsible gene by phenotypic complementation of a clear zone negative mutant . An open reading frame (lcp) of 1,191 bp was identified, which was preceded by a putative signal sequence and restored the capability to form clear zones on natural rubber latex in the mutant . The putative translation product exhibited strong homologies (50% aa identity) to a putative secreted protein from Streptomyces coelicolor strain A3(2), another clear zone forming strain . Heterologous expression of lcp of Streptomyces sp . strain K30 in Streptomyces lividans strain TK23 enabled the latter to form clear zones on latex-overlay agar plates and to accumulate a degradation product of about 12 kDa containing aldehyde groups . Two ORFs putatively encoding a heterodimeric molybdenum hydroxylase (oxiAB) were identified downstream of lcp in Streptomyces sp . strain K30 strain which exerted a positive effect on clear zone formation and enabled the strain to oxidize the resulting aldehydes . Heterologous expression of a fragment harboring lcp plus oxiAB in S . lividans TK23 resulted in accumulation of aldehydes only in the presence of 10 mM tungstate . Determination of protein content during cultivation on poly(cis-1,4-isoprene) revealed an increase of the cellular protein, and gel permeation chromatography analysis indicated a shift of the molecular weight distribution of the rubber to lower values in the transgenic S . lividans strains and in the wild type, thus confirming utilization and degradation of rubber . Therefore, for the first time, genes responsible for clear zone formation on natural rubber latex and synthetic cis-1,4-polyisoprene degradation in Gram-positive bacteria were identified and characterized. J Mol Med . 2005 Jan 6; {Epub ahead of print} Reading the viral signature by Toll-like receptors and other pattern recognition receptors; Mogensen TH et al.; Successful host defense against viral infections relies on early production of type I interferon (IFN) and subsequent activation of a cellular cytotoxic response . The acute IFN and inflammatory response against virus infections is mediated by cellular pattern-recognition receptors (PRRs) that recognize specific molecular structures on viral particles or products of viral replication . Toll-like receptors (TLRs) constitute a class of membrane-bound PRRs capable of detecting microbial infections . While TLR2 and TLR4, which were first identified to recognize Gram-positive and Gram-negative bacteria, respectively, sense specific viral proteins on the cell surface, TLRs 3, 7, 8, and 9 serve as receptors for viral nucleic acids in endosomic compartments . In addition to TLRs, cells express cytoplasmic PRRs such as the RNA helicase retinoic acid inducible gene I and the kinase double-stranded RNA-activated protein kinase R, both of which sense dsRNA, a characteristic signature of viral replication, and initiate a protective cellular response . Here we review the recent progress in our understanding of PRRs and viral infections and discuss the molecular and cellular responses evoked by virus-activated PRRs . Finally, we look into what is currently known about the role of PRRs in viral infections in vivo. Zhongguo Shi Yan Xue Ye Xue Za Zhi, 2004 Dec, 12(6), 861 - 5 {Analysis of the distribution characteristics and drug sensitivity of infection pathogens in hematology department ward.}; Li DJ et al.; The objective of this study was to investigate clinical distribution characteristics and drug sensitivity of infection pathogens in hematology department wards of our hospital during recent two years . The isolation and cultivation of pathogens from samples were performed by routine methods . Drug sensitivity tests of bacteria were performed by Kirby-Bauer method . Drug sensitivity tests of fungi were performed by ATBFUNGUS Drug sensitivity strips . The results showed that 102 strains of pathogens were isolated from all detected samples . The composition ratio of Gram-positive bacteria, Gram-negative bacteria and fungi was 42.2%, 34.3%, 3.5%, respectively . 58.8% of pathogens were isolated from samples of malignant hematopathy patients . 27.5% were isolated from samples of the patients with fever of unknown origin (FUO) . 51.0% of pathogens were isolated from samples of the patients who suffered from agranulocytosis or leucocytopenia . Isolated fungi were mostly sensitive to anti-fungal drugs . G+ bacteria were most sensitive to vancomycin . G- bacteria were most sensitive to imipenem . Most bacteria were resistant to multiple antibiotics . It is concluded that the infection in hematology department wards was related with many conditions, such as weakened resistance of patients, leucocytopenia or agranulocytosis, tumor loading, etc . The prompt microbiologcal examination and drug sensitivity tests are important to rationally select antibiotics, reduce infection incidence and mortality rate, and decrease the occurence of drug resistant strains. J Bacteriol, 2005 Jan, 187(2), 716 - 28 The bldC Developmental Locus of Streptomyces coelicolor Encodes a Member of a Family of Small DNA-Binding Proteins Related to the DNA-Binding Domains of the MerR Family; Hunt AC et al.; The bldC locus, required for formation of aerial hyphae in Streptomyces coelicolor, was localized by map-based cloning to the overlap between cosmids D17 and D25 of a minimal ordered library . Subcloning and sequencing showed that bldC encodes a member of a previously unrecognized family of small (58- to 78-residue) DNA-binding proteins, related to the DNA-binding domains of the MerR family of transcriptional activators . BldC family members are found in a wide range of gram-positive and gram-negative bacteria . Constructed DeltabldC mutants were defective in differentiation and antibiotic production . They failed to form an aerial mycelium on minimal medium and showed severe delays in aerial mycelium formation on rich medium . In addition, they failed to produce the polyketide antibiotic actinorhodin, and bldC was shown to be required for normal and sustained transcription of the pathway-specific activator gene actII-orf4 . Although DeltabldC mutants produced the tripyrrole antibiotic undecylprodigiosin, transcripts of the pathway-specific activator gene (redD) were reduced to almost undetectable levels after 48 h in the bldC mutant, in contrast to the bldC(+) parent strain in which redD transcription continued during aerial mycelium formation and sporulation . This suggests that bldC may be required for maintenance of redD transcription during differentiation . bldC is expressed from a single promoter . S1 nuclease protection assays and immunoblotting showed that bldC is constitutively expressed and that transcription of bldC does not depend on any of the other known bld genes . The bldC18 mutation that originally defined the locus causes a Y49C substitution that results in instability of the protein. J Pineal Res, 2005 Jan, 38(1), 17 - 26 Reactive changes of interstitial glia and pinealocytes in the rat pineal gland challenged with cell wall components from gram-positive and -negative bacteria; Jiang-Shieh YF et al.; Lipopolysaccharide (LPS), the major proinflammatory component of gram-negative bacteria, is well known to induce sepsis and microglial activation in the CNS . On the contrary, the effect of products from gram-positive bacteria especially in areas devoid of blood-brain barrier remains to be explored . In the present study, a panel of antibodies, namely, OX-6, OX-42 and ED-1 was used to study the response of microglia/macrophages in the pineal gland of rats given an intravenous LPS or lipoteichoic acid (LTA) . These antibodies recognize MHC class II antigens, complement type 3 receptors and unknown lysosomal proteins in macrophages, respectively . In rats given LPS (50 mug/kg) injection and killed 48 h later, the cell density and immunoexpression of OX-6, OX-42 and ED-1 in pineal microglia/macrophages were markedly increased . In rats receiving a high dose (20 mg/kg) of LTA, OX-42 and OX-6, immunoreactivities in pineal microglia/macrophages were also enhanced, but that of ED-1 was not . In addition, both bacterial toxins induced an increase in astrocytic profiles labelled by glial fibrillary acid protein . An interesting feature following LPS or LTA treatment was the lowering effect on serum melatonin, enhanced serotonin immunolabelling and cellular vacuolation as studied by electron microscopy in pinealocytes . The LPS- or LTA-induced vacuoles appeared to originate from the granular endoplasmic reticulum as well as the Golgi saccules . The present results suggest that LPS and LTA could induce immune responses of microglia/macrophages and astroglial activation in the pineal gland . Furthermore, the metabolic and secretory activity of pinealocytes was modified by products from both gram-positive and -negative bacteria. J Biol Chem . 2004 Dec 21; {Epub ahead of print} Evidence that the streptomyces developmental protein WhiD, a member of the WhiB family, binds a {4Fe-4S} cluster; Jakimowicz P et al.; WhiD is required for the late stages of sporulation in the Gram-positive bacterium Streptomyces coelicolor . WhiD is a member of the WhiB-like family of putative transcription factors, which are present throughout the actinomycetes, but absent from other organisms . This family of proteins has four near-invariant cysteines, suggesting that these residues might act as ligands for a metal cofactor . Over-expressed WhiD, purified from E . coli, contained sub-stoichiometric amounts of iron and had an absorption spectrum characteristic of a {2Fe-2S} cluster . After Fe-S cluster reconstitution under anaerobic conditions, WhiD contained ~4 Fe atoms per monomer and similar amounts of sulphide ion, and gave an absorption spectrum characteristic of a {4Fe-4S} cluster . Reconstituted WhiD gave no EPR signal as prepared, but, after reduction with dithionite, gave an EPR signal (g ~ 2.06, 1.94), consistent with a one electron reduction of a {4Fe-4S}2+ cluster to a {4Fe-4S}1+ state with electron spin of S=(1/2) . The anaerobically reconstituted {4Fe-4S} cluster was oxygen sensitive . Upon exposure to air, absorption at 410 and 505 nm first increased and then showed a steady decrease with time until the protein was colourless in the near UV/visible region . These changes are consistent with an oxygen-induced change from a {4Fe-4S} to a {2Fe-2S} cluster, followed by complete loss of cluster from the protein . Each of the four conserved cysteine residues, C23, C53, C56 and C62, was individually essential for WhiD function in vivo. Syst Appl Microbiol, 2004 Nov, 27(6), 636 - 45 Deinococcus frigens sp . nov., Deinococcus saxicola sp . nov., and Deinococcus marmoris sp . nov., low temperature and draught-tolerating, UV-resistant bacteria from continental Antarctica; Hirsch P et al.; Six Gram-positive, non-motile, UV- and draught-tolerant bacteria were isolated from antarctic soil and rock samples . The pink to orange cocci grew well on oligotrophic medium PYGV (pH 7.5) at 9-18 degrees C . They tolerated 0-10% NaCl, were aerobic to facultatively anaerobic and contained ornithine in their cell wall (type A3beta, Orn-Gly2) . The lipid profiles of four strains were found to be typical for those of D . radiodurans . Major fatty acids were 16:1cis9, 15:1cis9, 17:1cis9 and i17:1cis9, the respiratory quinone of three strains was MK-8 . Comparative 16S rDNA gene sequencing revealed phylogenetic relationships to the Deinococcus clade, especially to D . radiopugnans . The levels of 16S rRNA gene sequence similarity and DNA-DNA hybridisation data showed the six isolates represented new taxa . Phenotypic properties supported the description of three new species which were different from the eight known Deinococcus species and particularly from D . radiopugnans . Soil isolate AA-692T (DSM 12807T) is the type strain of Deinococcus frigens sp . nov., with AA-752 (DSM 15993) and AA-829 (DSM 15994) as additional strains from soil . The endolithic isolate AA-1444T, Deinococcus saxicola sp . nov., (DSM 15974T) came from antarctic sandstone, and Deinococcus marmoris sp . nov . (isolate AA-63T {DSM 12784T}) as well as AA-69 (DSM 15951) were isolated from antarctic marble. Dis Aquat Organ, 2004 Nov 4, 61(3), 263 - 8 Response of American lobsters Homarus americanus to infection with a field isolate of Aerococcus viridans var . homari (Gaffkemia): survival and haematology; Battison AL et al.; American lobsters Homarus americanus were inoculated with a field isolate of the Gram-positive bacterium Aerococcus viridans var . homari, causative agent of gaffkemia, at 1 x 10(6), 1 x 10(4) or 1 x 10(2) colony forming units (CFU) kg(-1) or with sterile 3% NaCl and maintained at 10 or 15 degrees C until they died or were euthanised . Progression of disease in individual animals was monitored daily by total haemocyte count (THC) and haemolymph culture . Post-mortem examinations were performed on all lobsters . Effects of both ambient temperature and infective dose on survival time were observed . Marked bacteraemia occurred in all mortalities . Haemocytopenia (THC < 10 x 10(9) cells l(-1)) preceded death in most, but not all, mortalities. Inorg Chem, 2004 Dec 27, 43(26), 8538 - 46 Synthesis, solution behavior, thermal stability, and biological activity of an Fe(III) complex of an artificial siderophore with intramolecular hydrogen bonding networks; Matsumoto K et al.; Previously, an artificial siderophore complex, the iron(III) complex with tris{2-{(N-acetyl-N-hydroxy)glycylamino}ethyl}amine (TAGE), was constructed in order to understand the effect of intramolecular hydrogen bonding interaction on the siderophore function, and its structural characterization in the solid state was reported (Inorg . Chem . 2001, 40, 190) . In this paper, the solution behavior of the M(III)-TAGE (M = Fe, Ga) system has been investigated using (1)H NMR, UV-vis, and FAB mass spectroscopies in efforts to characterize the biological implication of hydrogen bonding networks between the amide hydrogens and coordinating aminohydroxy oxygens of the complex . The temperature dependence of (1)H NMR spectra for Ga(III) complex of TAGE indicates that hydrogen bonding networks are maintained in polar solvents such as DMSO-d(6) and D(2)O . The UV-vis spectra of the Fe(III)-TAGE system under various pH conditions have shown that TAGE forms a tris(hydroxamato)iron(III) complex in an aqueous solution in the pH range 4-8 . By contrast, tris{2-{(N-acetyl-N-hydroxy)propylamido}ethyl}amine (TAPE; TAGE analogue that is difficult to form intramolecular hydrogen bonding networks), which has been synthesized as a comparison of TAGE, forms both of bis- and tris(hydroxamato)iron(III) complexes in the same pH range . Both the stability constants (log beta(FeTAGE) = 28.6; beta(FeTAGE) = {Fe(III)TAGE}/({Fe(3+)}{TAGE(3)(-)})) and pM (-log{Fe(3+)}) value for Fe(III)TAGE (pM 25) are comparable to those of a natural siderophore ferrichrome (log beta = 29.1 and pM 25.2) . The kinetic study of the TAGE-Fe(III) system has given the following rate constants: the rate of the ligand exchange reaction between Fe(III)TAGE and EDTA is 6.7 x 10(-4) s(-1), and the removal rates of iron from diferric bovine plasma transferrin by TAGE are 2.8 x 10(-2) and 6.0 x 10(-3) min(-1) . These values are also comparable to those of a natural siderophore desferrioxamine B under the same conditions . In a biological activity experiment, TAGE has promoted the growth of the siderophore-auxotroph Gram-positive bacterium Microbacterium flavescens, suggesting that TAGE mimics the activity of ferrichrome . These results indicate that the artificial siderophore with intramolecular hydrogen bonding networks, TAGE, is a good structural and functional model for a natural ferrichrome. Nippon Rinsho, 2004 Dec, 62(12), 2184 - 8 {The epidemiology of SIRS.sepsis in Japan}; Shibata K et al.; We outline the epidemiology of the systemic inflammatory response syndrome (SIRS), sepsis and severe sepsis among intensive care unit (ICU) patients in Japan . One survey conducted in the ICU of a university hospital suggested that the prevalence of SIRS reached 84% among all ICU patients, and that about 8% of patients with SIRS progressed to severe sepsis . Gram-positive cocci were the most common isolates (58%) from ICU patients with sepsis . Moreover, a graded severity was noted from SIRS to sepsis and severe sepsis, with total in-hospital mortality of 6%, 20% and 63%, respectively . Thus, every effort should be made to decrease the risk of sepsis in ICU patients. Pneumologie, 2004 Dec, 58(12), 850 - 3 {Pneumonia induced by nocardia -- a case report}; Frickmann H et al.; We describe a case of lung manifestation of nocardiosis with upper lobe shrinking of the right lung in a 45 year old patient without evident signs of an immuno-compromising illness . The patient came to the hospital in a reduced general state of health with severe cough, red and brown sputum and exertional dyspnoea . X-ray pictures of the thorax showed inflammatory infiltration and shrinking of the upper left lobe of the right lung . Gram-positive, branching rods were detected in the patient's bronchial secretion with the microscope and in cultures . Nocoardia transvalensis was identified via polymerase chain reaction (PCR) . The antibiotic therapy was planned according to the bacterial resistance pattern . Imipenem was administered for 5 weeks and Amikacin was added for 3 weeks in the 3 (rd) week of therapy . The patient left the hospital in a good general state of health . There was no relapse. Chest, 2004 Dec, 126(6), 1954 - 61 99mTc-labeled interleukin-8 for scintigraphic detection of pulmonary infections; Rennen HJ et al.; BACKGROUND: Interleukin (IL)-8 is a chemotactic cytokine that binds with high affinity to receptors on neutrophils . Previously we showed that (99m)Tc-labeled IL-8 is highly suitable for scintigraphic imaging in rabbit models of IM infection and of colitis . STUDY DESIGN: (99m)Tc-labeled IL-8 was tested for its potential to image pulmonary infection in three experimental rabbit models: aspergillosis in immunocompromised rabbits, pneumococcal (Gram-positive) pneumonia, and Escherichia coli-induced (Gram-negative) pneumonia in immunocompetent rabbits (four rabbits in each group) . A derivative of hydrazinonicotinamide was used as bifunctional coupling agent to label IL-8 with (99m)Tc . Biodistribution of (99m)Tc IL-8 was determined both by gamma-camera imaging and by counting dissected tissues at 6 h after injection . RESULTS: (99m)Tc IL-8 enabled early (within 2 h after injection) and excellent visualization of localization and extent of pulmonary infection in each of the three experimental models of pulmonary infection . Uptake of (99m)Tc IL-8 in the infected lung and the contralateral lung was (in percentage of the injected dose per gram of tissue +/- SEM) at 6 h after injection 0.63 +/- 0.12 and 0.12 +/- 0.02 (aspergillosis), 0.89 +/- 0.04 and 0.44 +/- 0.04 (pneumococcal pneumonia), and 1.53 +/- 0.12 and 0.36 +/- 0.06 (E coli pneumonia), respectively . In the E coli model, uptake of (99m)Tc IL-8 in the focus of infection even exceeded uptake in the kidneys, the main clearing organs . CONCLUSION: (99m)Tc IL-8 offers many advantages over the conventionally used radiopharmaceuticals to image pulmonary infection, (67)Ga citrate and radiolabeled leukocytes, ie, rapid and easy preparation, short time span between injection and imaging, low radiation burden and, most importantly, clear delineation of the infectious foci. Microbes Infect, 2004 Dec, 6(15), 1382 - 7 Toll-like receptors: linking innate and adaptive immunity; Pasare C et al.; Detection of and response to microbial infections by the immune system depends largely on a family of pattern-recognition receptors called Toll-like receptors (TLRs) . These receptors recognize conserved molecular products derived from various classes of pathogens, including Gram-positive and -negative bacteria, DNA and RNA viruses, fungi and protozoa . Recognition of ligands by TLRs leads to a series of signaling events resulting in induction of acute responses necessary to kill the pathogen . TLRs are also responsible for the induction of dendritic cell maturation, which is responsible and necessary for initiation of adaptive immune responses . Although TLRs control induction of adaptive immunity, it is not clear at this point how responses are appropriately tailored by individual TLRs to the advantage of the host. Glia . 2004 Dec 8; {Epub ahead of print} Toll-like receptor 2 (TLR2) is pivotal for recognition of S . aureus peptidoglycan but not intact bacteria by microglia; Kielian T et al.; Toll-like receptor 2 (TLR2) is a pattern recognition receptor that plays an important role in enabling cells of the innate immune system to recognize conserved structural motifs on a wide array of pathogens including gram-positive bacteria . Although microglia have recently been shown to express TLR2, the functional significance of this receptor in mediating microglial activation remains unknown . To ascertain the importance of TLR2 in microglial responses to S . aureus and its cell wall product peptidoglycan (PGN), we evaluated primary microglia from TLR2 knockout (KO) and wild-type (WT) mice . TLR2 was found to play a pivotal role in PGN recognition and subsequent activation in primary microglia, as demonstrated by the attenuated expression of TNF-alpha, IL-12 p40, MIP-2, and MCP-1 in PGN-treated TLR2 KO microglia compared with WT cells . In contrast, the responses of TLR2 KO and WT microglia to S . aureus were qualitatively similar, indicating that alternative receptors are responsible for recognizing intact bacteria . Microarray analysis confirmed that TLR2 plays a central role in PGN recognition by primary microglia . The expression of MyD88, a central adapter molecule in TLR-dependent signaling, was similar in both TLR2 KO and WT microglia, suggesting that the defect in PGN recognition by the former is not due to alterations in this key signaling intermediate . These findings reveal the complex nature of gram-positive bacterial recognition by microglia, which occurs, in part, through engagement of TLR2 . (c) 2004 Wiley-Liss, Inc. J Immunol, 2004 Dec 15, 173(12), 7339 - 48 Peptidoglycan molecular requirements allowing detection by the Drosophila immune deficiency pathway; Stenbak CR et al.; Innate immune recognition of microbes is a complex process that can be influenced by both the host and the microbe . Drosophila uses two distinct immune signaling pathways, the Toll and immune deficiency (Imd) pathways, to respond to different classes of microbes . The Toll pathway is predominantly activated by Gram-positive bacteria and fungi, while the Imd pathway is primarily activated by Gram-negative bacteria . Recent work has suggested that this differential activation is achieved through peptidoglycan recognition protein (PGRP)-mediated recognition of specific forms of peptidoglycan (PG) . In this study, we have further analyzed the specific PG molecular requirements for Imd activation through the pattern recognition receptor PGRP-LC in both cultured cell line and in flies . We found that two signatures of Gram-negative PG, the presence of diaminopimelic acid in the peptide bridge and a 1,6-anhydro form of N-acetylmuramic acid in the glycan chain, allow discrimination between Gram-negative and Gram-positive bacteria . Our results also point to a role for PG oligomerization in Imd activation, and we demonstrate that elements of both the sugar backbone and the peptide bridge of PG are required for optimum recognition . Altogether, these results indicate multiple requirements for efficient PG-mediated activation of the Imd pathway and demonstrate that PG is a complex immune elicitor. Diagn Microbiol Infect Dis, 2004 Dec, 50(4), 261 - 5 Release of daptomycin from polymethylmethacrylate beads in a continuous flow chamber; Hall EW et al.; Because daptomycin is active against Gram-positive cocci, it may be useful in the treatment and prevention of bone and joint infections when incorporated into polymethylmethacrylate (PMMA) . The release kinetics of daptomycin from PMMA were studied in a continuous flow chamber designed to simulate in vivo conditions . Three-millimeter beads containing 2.5%, 7.5%, and 15.0% daptomycin (weight daptomycin per weight PMMA) were individually placed in a chamber with 1 mL Krebs Ringer buffer flowing at 1 mL/hour . The majority of daptomycin was released in the first 24 hours . The mean peak concentrations were 13.4, 62.3, and 146.7 microg/mL; the mean AUC0-infinity were 30, 272, and 1204 h x microg/mL; and the mean percentages of daptomycin released were 6%, 18%, and 42% for the beads containing 2.5%, 7.5%, and 15.0% daptomycin, respectively . Daptomycin is released from PMMA in a continuous flow chamber at a rate similar to that previously determined by our laboratory for vancomycin. J Mol Biol, 2005 Jan 21, 345(3), 535 - 45 Structure and function of the antibiotic resistance-mediating methyltransferase AviRb from Streptomyces viridochromogenes; Mosbacher TG et al.; The emergence of antibiotic-resistant bacterial strains is a widespread problem in medical practice and drug design, and each case requires the elucidation of the underlying mechanism . AviRb from Streptomyces viridochromogenes methylates the 2'-O atom of U2479 of the 23S ribosomal RNA in Gram-positive bacteria and thus mediates resistance to the oligosaccharide (orthosomycin) antibiotic avilamycin . The structure of AviRb with and without bound cofactor S-adenosyl-L-methionine (AdoMet) was determined, showing that it is a homodimer belonging to the SpoU family within the SPOUT class of methyltransferases . The relationships within this class were analyzed in detail and, in addition, a novel fourth SpoU sequence fingerprint is proposed . Each subunit of AviRb consists of two domains . The N-terminal domain, being related to the ribosomal proteins L30 and L7Ae, is likely to bind RNA . The C-terminal domain is related to all SPOUT methyltransferases, and is responsible for AdoMet-binding, catalysis and dimerization . The cofactor binds at the characteristic knot of the polypeptide in an unusually bent conformation . The transferred methyl group points to a broad cleft formed with the L30-type domain of the other subunit . Measurements of mutant activity revealed four important residues responsible for catalysis and allowed the modeling of a complex between AviRb and the RNA target . The model includes a specificity pocket for uracil but does not contain a base for deprotonating the 2'-O atom of U2479 on methylation. Biotechnol Prog, 2004 Nov-Dec, 20(6), 1615 - 22 Microbial surfaces investigated using atomic force microscopy; Bolshakova AV et al.; This paper is dedicated to atomic force microscopy (AFM) as a progressive tool for imaging bacterial surfaces and probing their properties . The description of the technique is complemented by the explanation of the method's artifacts typical, in particular, for the imaging of bacterial cells . Sample preparation techniques are summarized in a separate section . Special attention is paid to the differences in imaging of gram-positive and gram-negative bacteria . Probing of mechanical properties, including elastic modulus, fragility, and adhesion of the cell walls is emphasized . The advantages of AFM in the studies of real-time cellular dynamical processes are illustrated by the experiment with the germination of spores. Intern Med, 2004 Oct, 43(10), 1000 - 4 A woman with infectious endocarditis caused by Abiotrophia defectiva; Hashimoto T et al.; A 74-year-old woman with recurrent fever and multiple joint pain was admitted to Hokkaido University Hospital . Trans-esophageal echocardiogram revealed bacterial vegetation and destruction of the aortic valve . Although few bacteria grew in regular blood agar, Gram-positive coccobacillus was specifically grown in chocolate blood agar and Brucella agar, and it was identified to be Abiotrophia defectiva . Infectious endocarditis caused by Abiotrophia defectiva was diagnosed, she was treated with diuretics, penicillin G and gentamicin, and she immediately improved . Infectious diseases caused by Abiotrophia defectiva are extremely rare, and identification of this pathogen is important, as its bacterial characteristics require proper attention. J Antimicrob Chemother, 2005 Jan, 55(1), 90 - 4 Epub 2004 Dec 01. Treatment of long-term intravascular catheter-related bacteraemia with antibiotic lock: randomized, placebo-controlled trial; Rijnders BJ et al.; OBJECTIVES: The use of an antibiotic lock (AB-lock) for the treatment of catheter-related bloodstream infection (CRBSI) has been suggested, but randomized trials have never been performed . METHODS: A randomized, blinded, multicentre trial was set up to compare an AB-lock-containing vancomycin for Gram-positive or ceftazidime for Gram-negative bacteria-with placebo, in addition to parenteral AB therapy . We included only CRBSI from a long-term intravascular device (LTID) whether tunnelled or totally implanted . RESULTS: During 30 months, 174 patients with an LTID and bacteraemia were evaluated, of whom 85 had a CRBSI . Forty-six patients were included . Frequent reasons for exclusion were: catheter not vacant for >8-12 h/day for the AB-lock (n =10); yeast infection or mixed Gram-positive/negative infections (n =13); catheter removal preferred by the treating physician (n =7); and CRBSI <14 days after insertion or pocket/tunnel infection (n =10) . Forty-four patients met the criteria for modified intention-to-treat analysis . The primary endpoint was failure to cure the CRBSI or relapse with the same strain . On study day 180 by Kaplan-Meier analysis, this occurred in 33% (seven of 21) in the AB-lock arm and in 57% (13 of 23) in the placebo arm (hazard ratio 0.55, P =0.10) . A relapse with the same strain occurred in 9/23 with the placebo and 3/21 with the AB-lock (P =0.06) . CONCLUSION: Future studies should take into account the barriers to the use of AB-lock observed in this study . Most importantly, shorter lock dwell times and broader spectrum locks (e.g . antiseptic) should be investigated to target a larger patient population. Proc Natl Acad Sci U S A, 2004 Dec 7, 101(49), 17168 - 73 Epub 2004 Dec 7. Structural basis for peptidoglycan binding by peptidoglycan recognition proteins; Guan R et al.; Peptidoglycan (PGN) recognition proteins (PGRPs) are pattern-recognition receptors of the innate immune system that bind and, in some cases, hydrolyze bacterial PGNs . We determined the crystal structure, at 2.30-A resolution, of the C-terminal PGN-binding domain of human PGRP-Ialpha in complex with a muramyl tripeptide representing the core of lysine-type PGNs from Gram-positive bacteria . The peptide stem of the ligand is buried at the deep end of a long binding groove, with N-acetylmuramic acid situated in the middle of the groove, whose shallow end can accommodate a linked N-acetylglucosamine . Although most interactions are with the peptide, the glycan moiety also seems to be essential for specific recognition by PGRPs . Conservation of key PGN-contacting residues shows that all PGRPs employ this basic PGN-binding mode . The structure pinpoints variable residues that likely mediate discrimination between lysine- and diaminopimelic acid-type PGNs . We also propose a mechanism for PGN hydrolysis by Zn(2+)-containing PGRPs. Nihon Kokyuki Gakkai Zasshi, 2004 Oct, 42(10), 893 - 6 {A case of pulmonary nocardiosis with a polypoid lesion in a bronchus}; Morita H et al.; A 62-year-old woman was admitted with fever and bloody sputum . A mass shadow in the left S3 and obstruction of the left B3 were seen on a chest radiograph and CT . Obstructive pneumonia was suspected, and cefotiam and imipenem/cilastatin were administered . However, this treatment did not show adequate efficacy . Bronchoscopy demonstrated a yellowish-white polypoid lesion in the left B3, but histopathological findings with HE staining yielded no definite diagnosis . Subsequently, Nocardia asteroides was detected in sputum test . A sulfamethoxazole-trimethoprim combination and minocycline were administered, and the clinical findings improved . Gram-positive microfilaments were confirmed retrospectively in the pathologic specimen, and a diagnosis of pulmonary nocardiosis was made. Eur J Biochem, 2004 Nov, 271(22), 4361 - 5 The Thermoplasma acidophilum Lon protease has a Ser-Lys dyad active site; Besche H et al.; A gene with significant similarity to bacterial Lon proteases was identified during the sequencing of the genome of the thermoacidophilic archaeon Thermoplasma acidophilum . Protein sequence comparison revealed that Thermoplasma Lon protease (TaLon) is more similar to the LonB proteases restricted to Gram-positive bacteria than to the widely distributed bacterial LonA . However, the active site residues of the protease and ATPase domain are highly conserved in all Lon proteases . Using site-directed mutagenesis we show here that TaLon and EcLon, and probably all other Lon proteases, contain a Ser-Lys dyad active site . The TaLon active site mutants were fully assembled and, similar to TaLon wild-type, displayed an apparent molar mass of 430 kDa upon gelfiltration . This would be consistent with a hexameric complex and indeed electron micrographs of TaLon revealed ring-shaped particles, although of unknown symmetry . Comparison of the ATPase activity of Lon wild-type from Thermoplasma or Escherichia coli with respective protease active site mutants revealed differences in Km and V values . This suggests that in the course of protein degradation by wild-type Lon the protease domain might influence the activity of the ATPase domain. Biochem J . 2004 Nov 23; {Epub ahead of print} TraA and its N-terminal relaxase domain of the Gram-positive plasmid pIP501 show specific oriT binding and behave as dimers in solution; Kopec J et al.; TraA is the DNA relaxase encoded by the broad-host-range Gram-positive plasmid pIP501 . It is the second relaxase characterized from plasmids originating from Gram-positive organisms . TraA (654 amino acids) and the N-terminal domain (246 amino acids), termed TraAN 246, were expressed as 6xHis-tagged fusions and purified . Small-angle X-ray scattering and chemical cross-linking proved that TraAN 246 and TraA form dimers in solution . Both proteins revealed oriT pIP501 cleavage activity on supercoiled plasmid DNA in vitro . oriT binding was demonstrated by electrophoretic mobility shift assays . Radiolabeled oligonucleotides covering different parts of oriT pIP501 were subjected to binding with TraA and TraAN 246 . The K D of the protein-DNA complex encompassing the inverted repeat (IR), the nick site and additional 7 bases, was found to be 55 nM for TraA, and 26 nM for TraAN 246 . The unfolding of both protein constructs was monitored by measuring the change in the circular dichroism (CD) signal at 220 nm upon temperature change . The unfolding transition of both proteins occurred at around 42 degrees C . CD spectra measured at 20 degrees C showed 30 % alpha-helix and 13 % beta-sheet for TraA and 27 % alpha-helix and 18 % beta-sheet content for the truncated protein . Upon DNA-binding an enhanced secondary structure content and increased thermal stability was observed for the TraAN 246 protein suggesting an induced-fit mechanism for the formation of the specific relaxase-oriT complex. Mikrobiol Z, 2004 Sep-Oct, 66(5), 62 - 7 {Antibiotic and phytotoxic properties of some Aspergillus parvulus Smith strains} {An unusual case of hepatic abscess} Jermini I, Luethy R, Gubler J. Medizinische Klinik, Stadtspital Triemli ZurichRight upper quadrant pain with chronic malaise and fever can be the clinical manifestation of a hepatic infection . An uncommon cause for the disease is hepatic actinomycosis . Actinomycosis was common in the preantibiotic era but is less frequent nowadays; consequently its timely recognition has become more difficult . The clinical and radiological findings often resemble other inflammatory and neoplastic lesions . We report a case of a mixed anaerobic liver abscess including fusobacteria and actinomycetes, without apparent predisposing factor . The diagnosis was obtained by CT-guided percutaneous aspiration of the hepatic mass, where microscopy revealed the presence of fusiform gramnegative bacteria and gram-positive branching filamentous rods consistent with Actinomyces species . The latter did not grow in culture, while the gram negatives were identified as fusobacterium nucleatum . The diagnosis of actinomycosis of the liver is confirmed in only a minority of cases by culture . The disease is usually treated with an extended course of antibiotics . Penicillin is the preferred choice. Ned Tijdschr Geneeskd, 2004 Oct 23, 148(43), 2115 - 21 {New fluoroquinolones: levofloxacin, moxifloxacin and gatifloxacin}; Hoepelman I; Since their introduction in the seventies, the fluoroquinolones have gained an important place in the treatment of Gram-negative infections . Modifications in their structure have led to a number of newer agents with improved activity against Gram-positive microorganisms . Among these new agents are gatifloxacin, levofloxacin and moxifloxacin, which have been registered in The Netherlands . These new fluoroquinolones are promoted by the manufacturers for use in respiratory tract infections, notably in community-acquired pneumonia . Studies have shown that these agents may be somewhat more effective than the current antibiotics in use in The Netherlands for community-acquired pneumonia . Given the fact that penicillin resistance among pneumococci hardly occurs in The Netherlands and the severe risk of the development of resistance in both Gram-positive and Gram-negative microorganisms when the new fluoroquinolones are used, their area of indication would currently seem to be limited. Eur Arch Otorhinolaryngol . 2004 Nov 12; {Epub ahead of print} Actinomycosis of the middle turbinate: an unusual cause of nasal obstruction; Ozcan C et al.; Actinomycosis is a rare chronic infectious disease caused by Actinomyces israelii, which is an anaerobic filamentous, gram-positive saprophyte organism of the oral cavity . Historically, these bacteria were known as fungi because of their light microscopic appearance . Actinomycosis consists of three different forms: cervico-facial (the most common form), abdominal and pulmonothoracic . It commonly involves the head and neck region including the mandible, paranasal sinuses, lacrimal gland, parotid gland and orbit . Poor oral hygiene and dental diseases have been known to be the source of actinomycosis . Actinomycosis is diagnosed with positive culture or detecting actinomyces colonies and sulfur granules in histopathologic specimens . The treatment of choice is surgical excision of the lesion and long-term penicillin therapy . Actinomycosis of the internal nose is extremely rare . There was only one nasal septum actinomycosis reported in the English literature, but there was no lateral nasal wall actiomycosis regarding the turbinate . Therefore, actinomycosis should not be overlooked for the differential diagnosis of intra-nasal lesions for the initiation of appropriate and early treatment. J Bacteriol, 2004 Dec, 186(23), 8153 - 5 A self-splicing group I intron in DNA polymerase genes of T7-like bacteriophages; Bonocora RP et al.; Group I introns are inserted into genes of a wide variety of bacteriophages of gram-positive bacteria . However, among the phages of enteric and other gram-negative proteobacteria, introns have been encountered only in phage T4 and several of its close relatives . Here we report the insertion of a self-splicing group I intron in the coding sequence of the DNA polymerase genes of PhiI and W31, phages that are closely related to T7 . The introns belong to subgroup IA2 and both contain an open reading frame, inserted into structural element P6a, encoding a protein belonging to the HNH family of homing endonucleases . The introns splice efficiently in vivo and self-splice in vitro under mild conditions of ionic strength and temperature . We conclude that there is no barrier for maintenance of group I introns in phages of proteobacteria. Biochim Biophys Acta, 2004 Nov 11, 1694(1-3), 269 - 78 Protein sorting to the cell wall envelope of Gram-positive bacteria; Ton-That H et al.; The covalent anchoring of surface proteins to the cell wall envelope of Gram-positive bacteria occurs by a universal mechanism requiring sortases, extracellular transpeptidases that are positioned in the plasma membrane . Surface protein precursors are first initiated into the secretory pathway of Gram-positive bacteria via N-terminal signal peptides . C-terminal sorting signals of surface proteins, bearing an LPXTG motif or other recognition sequences, provide for sortase-mediated cleavage and acyl enzyme formation, a thioester linkage between the active site cysteine residue of sortase and the C-terminal carboxyl group of cleaved surface proteins . During cell wall anchoring, sortase acyl enzymes are resolved by the nucleophilic attack of peptidoglycan substrates, resulting in amide bond formation between the C-terminal end of surface proteins and peptidoglycan cross-bridges within the bacterial cell wall envelope . The genomes of Gram-positive bacteria encode multiple sortase genes . Recent evidence suggests that sortase enzymes catalyze protein anchoring reactions of multiple different substrate classes with different sorting signal motif sequences, protein linkage to unique cell wall anchor structures as well as protein polymerization leading to the formation of pili on the surface of Gram-positive bacteria. Microb Ecol, 2004 Aug, 48(2), 209 - 17 Epub 2004 Jun 10. Genetic diversity of bacterial communities of serpentine soil and of rhizosphere of the nickel-hyperaccumulator plant Alyssum bertolonii; Mengoni A et al.; Serpentine soils are characterized by high levels of heavy metals (Ni, Co, Cr), and low levels of important plant nutrients (P, Ca, N) . Because of these inhospitable edaphic conditions, serpentine soils are typically home to a very specialized flora including endemic species as the nickel hyperaccumulator Alyssum bertolonii . Although much is known about the serpentine flora, few researches have investigated the bacterial communities of serpentine areas . In the present study bacterial communities were sampled at various distances from A . bertolonii roots in three different serpentine areas and their genetic diversity was assessed by terminal restriction fragment length polymorphism (T-RFLP) analysis . The obtained results indicated the occurrence of a high genetic diversity and heterogeneity of the bacterial communities present in the different serpentine areas . Moreover, TRFs (terminal restriction fragments) common to all the investigated A . bertolonii rhizosphere samples were found . A new cloning strategy was applied to 27 TRFs that were sequenced and taxonomically interpreted as mainly belonging to Gram-positive and alpha-Proteobacteria representatives . In particular, cloned TRFs which discriminated between rhizosphere and soil samples were mainly interpreted as belonging to Proteobacteria representatives. Int J Syst Evol Microbiol, 2004 Nov, 54(Pt 6), 2013 - 7 Exiguobacterium oxidotolerans sp . nov., a novel alkaliphile exhibiting high catalase activity; Yumoto I et al.; A novel alkaliphile was isolated from a drain of a fish processing plant . The isolate grew at a pH range of 7-10 . Cells were Gram-positive, facultatively aerobic, motile rods with peritrichous flagella . Colonies were orange or yellow in colour . Catalase and oxidase reactions were positive . The isolate grew in 0-12 % NaCl but not above 15 % NaCl . Its cell extract exhibited 567 times higher catalase activity than an Escherichia coli cell extract . The major cellular fatty acids were iso-C(13 : 0), anteiso-C(13 : 0), iso-C(15 : 0), iso-C(16 : 0), iso-C(17 : 0), anteiso-C(17 : 0) and iso-C(17 : 1) . Its DNA G+C content was 46.7 mol% . Phylogenetic analysis based on 16S rRNA gene sequencing and chemotaxonomic data indicated that strain T-2-2(T) is a member of the genus Exiguobacterium . DNA-DNA hybridization revealed a low relatedness of the isolate to several phylogenetic neighbours (less than 25 %) . On the basis of phenotypic characteristics, phylogenetic data and DNA-DNA relatedness data, the isolate merits classification as a novel species, for which the name Exiguobacterium oxidotolerans sp . nov . is proposed . The type strain is T-2-2(T) (=JCM 12280(T)=NCIMB 13980(T)). Int J Syst Evol Microbiol, 2004 Nov, 54(Pt 6), 1987 - 90 Agromyces ulmi sp . nov., a xylanolytic bacterium isolated from Ulmus nigra in Spain; Rivas R et al.; Two xylan-degrading bacterial strains were isolated from a decayed Ulmus nigra tree in Spain . The isolates were Gram-positive, non-motile, aerobic and formed substrate mycelium which fragmented into irregular rods . 16S rRNA gene sequence analysis indicated that the isolates form a separate branch within the genus Agromyces phylogenetic cluster, with Agromyces mediolanus DSM 20152(T) being their closest relative (97.7 and 97.6 % sequence similarity) . Catalase, nitrate reduction and urease tests differentiated these strains from A . mediolanus . Cell-wall peptidoglycan composition, major menaquinone, predominant fatty acids and phospholipid pattern were typical of the genus Agromyces . The DNA G+C content determined for the type strain XIL01(T) was 72 mol% . Based on the data presented, a novel species Agromyces ulmi sp . nov . is proposed . The type strain is XIL01(T) (=LMG 21954(T)=DSM 15747(T)). Int J Syst Evol Microbiol, 2004 Nov, 54(Pt 6), 1975 - 80 Janibacter melonis sp . nov., isolated from abnormally spoiled oriental melon in Korea; Yoon JH et al.; Two Gram-positive bacterial strains, CM2104(T) and CM2110, isolated from the inner part of abnormally spoiled oriental melon (Cucumis melo) in Korea, were subjected to a polyphasic taxonomic study . The cell-wall peptidoglycan of strains CM2104(T) and CM2110 contained meso-diaminopimelic acid as the diagnostic diamino acid . The predominant menaquinone was MK-8(H(4)) . The major fatty acids detected in the two strains were iso-C(16 : 0), C(17 : 1)omega8c and C(18 : 1)omega9c or C(17 : 0) . The DNA G+C content of the two strains was 73 mol% . Phylogenetic analysis based on 16S rRNA gene sequences showed that the strains formed a coherent cluster with a clade comprising two Janibacter species, Janibacter limosus and Janibacter terrae . Strains CM2104(T) and CM2110 exhibited a 16S rRNA gene sequence similarity value of 99.7 % and a mean DNA-DNA relatedness level of 89 % . Strains CM2104(T) and CM2110 showed 16S rRNA gene sequence similarity levels of 97.8-98.4 % to the type strains of J . limosus and J . terrae . DNA-DNA relatedness between strains CM2104(T) and CM2110 and the type strains of these two Janibacter species was 7-11 % . On the basis of the phenotypic and phylogenetic data and genomic distinctiveness, strains CM2104(T) and CM2110 should be placed within the genus Janibacter as members of a novel species, for which the name Janibacter melonis sp . nov . is proposed . The type strain is CM2104(T) (=KCTC 9987(T)=DSM 16063(T)=JCM 12321(T)). Biochim Biophys Acta, 2004 Nov 17, 1667(1), 47 - 55 Identification of a cation-specific channel (TipA) in the cell wall of the gram-positive mycolata Tsukamurella inchonensis: the gene of the channel-forming protein is identical to mspA of Mycobacterium smegmatis and mppA of Mycobacterium phlei; Dorner U et al.; Detergent extracts of whole cells of the Gram-positive bacterium Tsukamurella inchonensis ATCC 700082, which belongs to the mycolata, were studied for the presence of ion-permeable channels using lipid bilayer experiments . One channel with a conductance of about 4.5 nS in 1 M KCl was identified in the extracts . The channel-forming protein was purified to homogeneity by preparative SDS-PAGE . The protein responsible for channel-forming activity had an apparent molecular mass of about 33 kDa as judged by SDS-PAGE . Interestingly, the protein showed cross-reactivity with polyclonal antibodies raised against a polypeptide derived from MspA of Mycobacterium smegmatis similarly as the cell wall channel of Mycobacterium phlei . Primers derived from mspA were used to clone and sequence the gene of the cell wall channels of T . inchonensis (named tipA for T . inchonensis porin A) and M . phlei (named mppA for M . phlei porin A) . Surprisingly, both genes, tipA and mppA, were found to be identical to mspA of M . smegmatis, indicating that the genomes of T . inchonensis, M . phlei and M . smegmatis contain the same genes for the major cell wall channel . RT-PCR revealed that tipA is transcribed in T . inchonensis and mppA in M . phlei . The results suggest that despite a certain distance between the three organisms, their genomes contain the same gene coding for the major cell wall channel, with a molecular mass of 22 kDa for the monomer. Tunis Med, 2004 Aug, 82(8), 772 - 6 {Abdomino-pelvic actinomycosis . Two cases report}; Belkahla N et al.; Actinomycosis is a chronic, granulomatous, suppurative and fistulasing infection related to a gram-positive bacteria (actinomyces israeli) . Cervico-facial actinomycosis is the most common localization . The prevalence of abdomino-pelvic actinomycosis is increasing mainly with the increase of the use of intrauterin device . Its clinical presentation is variable and may mimic cancer or tuberculosis . The diagnosis of abdomino-pelvic actinomycosis is hard and most of the cases are detected during surgical exploration . We report two cases of abdomino-pelvic actinomycosis; in the first case, the disease was extended to the caecum and the abdominal wall . The skin biopsies made the diagnosis of actinomycosis, avoiding surgery . In the second case, the diagnosis of actinomycosis is made post operatively because of high suspicion of pelvic cancer . Through these two observations, we review pathogenesis of the disease, its clinical aspects and its diagnostic and therapeutic means. Microbiology, 2004 Nov, 150(Pt 11), 3783 - 95 Reconstruction of the evolutionary history of the LexA-binding sequence; Mazon G et al.; In recent years, the recognition sequence of the SOS repressor LexA protein has been identified for several bacterial clades, such as the Gram-positive, green non-sulfur bacteria and Cyanobacteria phyla, or the 'Alphaproteobacteria', 'Deltaproteobacteria' and 'Gammaproteobacteria' classes . Nevertheless, the evolutionary relationship among these sequences and the proteins that recognize them has not been analysed . Fibrobacter succinogenes is an anaerobic Gram-negative bacterium that branched from a common bacterial ancestor immediately before the Proteobacteria phylum . Taking advantage of its intermediate position in the phylogenetic tree, and in an effort to reconstruct the evolutionary history of LexA-binding sequences, the F . succinogenes lexA gene has been isolated and its product purified to identify its DNA recognition motif through electrophoretic mobility assays and footprinting experiments . After comparing the available LexA DNA-binding sequences with the F . succinogenes one, reported here, directed mutagenesis of the F . succinogenes LexA-binding sequence and phylogenetic analyses of LexA proteins have revealed the existence of two independent evolutionary lanes for the LexA recognition motif that emerged from the Gram-positive box: one generating the Cyanobacteria and 'Alphaproteobacteria' LexA-binding sequences, and the other giving rise to the F . succinogenes and Myxococcus xanthus ones, in a transitional step towards the current 'Gammaproteobacteria' LexA box . The contrast between the results reported here and the phylogenetic data available in the literature suggests that, some time after its emergence as a distinct bacterial class, the 'Alphaproteobacteria' lost its vertically received lexA gene, but received later through lateral gene transfer a new lexA gene belonging to either a cyanobacterium or a bacterial species closely related to this phylum . This constitutes the first report based on experimental evidence of lateral gene transfer in the evolution of a gene governing such a complex regulatory network as the bacterial SOS system. Microbiology, 2004 Nov, 150(Pt 11), 3749 - 61 Novel organization of genes in a phthalate degradation operon of Mycobacterium vanbaalenii PYR-1; Stingley RL et al.; Mycobacterium vanbaalenii PYR-1 is capable of degrading polycyclic aromatic hydrocarbons (PAHs) to ring cleavage metabolites . This study identified and characterized a putative phthalate degradation operon in the M . vanbaalenii PYR-1 genome . A putative regulatory protein (phtR) was encoded divergently with five tandem genes: phthalate dioxygenase large subunit (phtAa), small subunit (phtAb), phthalate dihydrodiol dehydrogenase (phtB), phthalate dioxygenase ferredoxin subunit (phtAc) and phthalate dioxygenase ferredoxin reductase (phtAd) . A 6.7 kb EcoRI fragment containing these genes was cloned into Escherichia coli and converted phthalate to 3,4-dihydroxyphthalate . Homologues to the operon region were detected in a number of PAH-degrading Mycobacterium spp . isolated from various geographical locations . The operon differs from those of other Gram-positive bacteria in both the placement and orientation of the regulatory gene . In addition, the M . vanbaalenii PYR-1 pht operon contains no decarboxylase gene and none was identified within a 37 kb region containing the operon . This study is the first report of a phthalate degradation operon in Mycobacterium spp. Appl Environ Microbiol, 2004 Nov, 70(11), 6920 - 6 Combined phospholipid biomarker-16S rRNA gene denaturing gradient gel electrophoresis analysis of bacterial diversity and physiological status in an intertidal microbial mat; Villanueva L et al.; A combined lipid biomarker-16S rRNA gene denaturing gradient gel electrophoresis analysis was used to monitor changes in the physiological status, biomass, and microbial composition of a microbial mat . In the morning hours, an increase in the biomass of layers containing a high density of phototrophs and a decrease in the growth rate in the deep layers were observed . The combined approach also revealed differences in major groups of microorganisms, including green nonsulfur, gram-positive, and heterotrophic bacteria. Appl Environ Microbiol, 2004 Nov, 70(11), 6488 - 94 Identification of a third msa gene in Renibacterium salmoninarum and the associated virulence phenotype; Rhodes LD et al.; Renibacterium salmoninarum, a gram-positive diplococcobacillus, causes bacterial kidney disease, a condition that can result in extensive morbidity and mortality among stocks of fish . An immunodominant extracellular protein, called major soluble antigen (MSA), is encoded by two identical genes, msa1 and msa2 . We found evidence for a third msa gene, msa3, which appears to be a duplication of msa1 . Unlike msa1 and msa2, msa3 is not present in all isolates of R . salmoninarum . The presence of the msa3 locus does not affect total MSA production in culture conditions . In a challenge study, isolates possessing the msa3 locus reduced median survival in juvenile chinook salmon (Oncorhynchus tshawytscha) by an average of 34% at doses of < or =10(5) cells per fish compared to isolates lacking the msa3 locus . In contrast, no difference in survival was observed at the highest dose, 10(6) cells per fish . The phenotype associated with the msa3 locus and its nonuniform distribution may contribute to observed differences in virulence among R . salmoninarum isolates. Ginekol Pol, 2004 Jun, 75(6), 439 - 44 {Procalcitonin and C-reactive protein as a markers of neonatal sepsis}; Kawczynski P et al.; OBJECTIVES: Sensitive, reliable and early parameters of bacterial infection are extremely valuable in diagnosis of nosocomial infections in neonatal intensive care unit . In this study procalcitonin (PCT) and C-reactive protein (CRP) were evaluated for their diagnostic relevance in neonatal late onset sepsis . DESIGN: Clinical study MATERIALS AND METHODS: We analysed inflammatory parameters in 48 newborn infants admitted to the Intensive Care Unit of Institute of Paediatrics in Lodz who suffered from nosocomial sepsis . They were sampled for PCT and CRP levels at the time of the onset of signs and 24 hours later . CRP was determined by an nephelometric method and PCT was determined by an immunoluminometric assay . RESULTS: At the onset of Gram negative sepsis 14 from 17 contaminated newborns had significantly increased CRP levels and 15 of them had increased levels of PCT After 24 hours 100% of them had elevated PCT and CRP levels . At the onset of Gram positive sepsis only 18 from 31 neonates with positive blood culture had increased CRP levels and 28 of them had elevated concentrations of PCT . This difference was statistically significant . After 24 hours 26 of them had elevated CRP and 100% had increased PCT concentrations--this difference was not significant . CONCLUSION: Measurement of procalcitonin concentrations may be useful for early diagnosis of late onset sepsis in neonates. Mikrobiologiia, 2004 Jul-Aug, 73(4), 479 - 85 {The mechanism of action of the extracellular bacteriolytic enzymes of Lysobacter sp . on gram-positive bacteria: the role of the cell wall anionic polymers of target bacteria} {Antibiotic properties of some species of genus Aspergillus mich} {No authors listed} Antibiotic activity screening of some species of genus Aspergillus (64 strains) to different test-organisms (gram positive, gram negative, phytopathogenic bacteria and yeasts) has been carried out . The researched strains A . parvulus, A . ochraceus, A . ustus, A . versicolor, A.fumigatus, A . niger, A . deflectus, A . clavatus were characterized by a wide spectrum of antibiotic activity; the strains A . flavipes, A . sydowii, A . terreus, A . flavus, A . nidulans, A . alliaceus, A . fluticulosus were characterized by a narrow spectrum of antibiotic activity . Fungistatic activity was shown for some strains of A . ochraceus, A . versicolor, A . ustus . The majority of investigated strains were characterized by the static action . The species A . parvulus which has not been investigated before is distinguished by a wide spectrum of antibiotic activities would be recognized most interesting for further investigation. Scand J Infect Dis, 2004, 36(10), 712 - 7 High versus standard dose vancomycin for osteomyelitis; Boffi El Amari E et al.; It is important to identify the optimal dosage and best method of infusion of parenteral vancomycin to be used over a several week period for the treatment of osteomyelitis . A retrospective study was undertaken to compare a high dose vancomycin treatment (HD: 40 mg/kg/d) with a standard dose treatment (SD: 20 mg/kg/d), and also to compare the modality of infusion using either intermittent vancomycin infusion (IVI) or continuous vancomycin infusion (CVI) . 89 patients with Gram-positive cocci osteomyelitis requiring vancomycin treatment were followed, and the outcome and therapeutic safety were compared . There were significantly more adverse drug reactions (acute renal failure) in the IVI subgroup (HD-IVI vs SD-IVI, p-value 0.007) . No cases of renal failure were found in the HD-CVI subgroup . The best outcome was found in the subgroup of patients who received HD-CVI (HD-CVI vs SD-IVI, overall log rank p-value 0.02) . HD-CVI treatment appears to provide an improved outcome with fewer adverse drug reactions. Am J Obstet Gynecol, 2004 Oct, 191(4), 1346 - 55 Toll-like receptor-2 and -4 in the chorioamniotic membranes in spontaneous labor at term and in preterm parturition that are associated with chorioamnionitis; Kim YM et al.; OBJECTIVE: The Toll-like receptor-2 and -4 recognize microbial products that are associated with gram-positive and gram-negative bacteria, respectively . This study examined Toll-like receptor-2 and -4 expression in fetal membranes in response to labor at term and histologic chorioamnionitis . STUDY DESIGN: Immunohistochemistry and real-time quantitative reverse transcriptase-polymerase chain reaction were used to examine the chorioamnion from patients with (1) preterm labor and intact membranes, (2) preterm premature rupture of membranes who were delivered < or =34 weeks of gestation, and (3) women at term with or without labor . All groups were stratified on the basis of the presence of histologic chorioamnionitis . RESULTS: Toll-like receptor-2 expression was significantly higher in patients with chorioamnionitis than in patients without this condition (all P < .05) . The Toll-like receptor-2 and -4 messenger RNA amounts were significantly higher in membranes from women at term with spontaneous labor than women who were not in labor ( P = .001 and .002, respectively) . Toll-like receptor-2 expression was polarized to the basal surface of amniotic epithelial cells in patients without chorioamnionitis, but this distribution was lost in the presence of inflammation . CONCLUSION: Spontaneous labor at term and preterm delivery with histologic chorioamnionitis, regardless of the membrane status (intact or ruptured), are associated with an increased expression of Toll-like receptor-2 and -4 in the chorioamniotic membranes . These observations have implications for understanding the biologic nature of innate immunity. Antimicrob Agents Chemother, 2004 Nov, 48(11), 4120 - 9 Chemical diversity of polyene macrolides produced by Streptomyces noursei ATCC 11455 and recombinant strain ERD44 with genetically altered polyketide synthase NysC; Bruheim P et al.; The gram-positive bacterium Streptomyces noursei ATCC 11455 produces a complex mixture of polyene macrolides generally termed nystatins . Although the structures for nystatins A(1) and A(3) have been reported, the identities of other components of the nystatin complex remain obscure . Analyses of the culture extract from the S . noursei wild type revealed the presence of several nystatin-related compounds for which chemical structures could be suggested on the basis of their molecular weights, their UV spectra, and knowledge of the nystatin biosynthetic pathway . Nuclear magnetic resonance (NMR) studies with one of these polyene macrolides identified it as a nystatin analogue containing a mycarose moiety at C-35 . A similar investigation was performed with the culture extract of the ERD44 mutant, which has a genetically altered polyketide synthase (PKS) NysC and which was previously shown to produce a heptaene nystatin analogue . The latter compound, tentatively named S44HP, and its derivative, which contains two deoxysugar moieties, were purified; and their structures were confirmed by NMR analysis . Nystatin analogues with an expanded macrolactone ring were also observed in the extract of the ERD44 mutant, suggesting that the altered PKS can "stutter" during the polyketide chain assembly . These data provide new insights into the biosynthesis of polyene macrolide antibiotics and the functionalities of PKSs and post-PKS modification enzymes. Hear Res, 2004 Nov, 197(1-2), 11 - 8 Minocycline attenuates gentamicin induced hair cell loss in neonatal cochlear cultures; Corbacella E et al.; Minocycline, a second-generation tetracycline antibiotic used against gram-negative and gram-positive bacteria, protects against a wide range of neurodegenerative disorders by inhibiting caspases, iNOS and the release of cytochrome c . Since aminoglycoside antibiotics damage sensory hair cells in the inner ear by activating caspase-mediated cell death pathways, we hypothesized that minocycline would protect against gentamicin (GM) ototoxicity . To test this hypothesis, postnatal day 3 (P3) rat, cochlear organotypic cultures were treated with GM alone or in combination with minocycline (10-500 microM) . Treatment with GM induced a dose-dependent loss of outer hair cells (OHC) and inner hair cells (IHC) . Addition of minocycline to the GM-treated cultures greatly reduced the amount of GM-induced hair cell damage in P3 cochlear cultures . The greatest protection was achieved with 100 microM of minocycline . Application of minocycline alone had no adverse effects on hair cell survival . The advantage of this combination therapy is that minocycline prevents GM-induced hair cell loss while helping to suppress the bacterial infection. Bioinformatics . 2004 Oct 22; {Epub ahead of print} PSORTb v.2.0: expanded prediction of bacterial protein subcellular localization and insights gained from comparative proteome analysis; Gardy JL et al.; MOTIVATION: PSORTb v.1.1 is the most precise bacterial localization prediction tool available . However the program's predictive coverage and recall are low and the method is only applicable to Gram-negative bacteria . The goals of the present work were: increase PSORTb's coverage while maintaining the existing precision level, expand it to include Gram-positive bacteria, and then carry out a comparative analysis of localization . RESULTS: An expanded database of proteins of known localization and new modules using frequent subsequence-based support vector machines were introduced into PSORTb v.2.0 . The program attains a precision of 96% for Gram-positive and Gram-negative bacteria and predictive coverage comparable to other tools for whole proteome analysis . We show that the proportion of proteins at each localization is remarkably consistent across species, even in species with varying proteome size . AVAILABILITY: Web-based version: Standalone version: Available through the website under GNU General Public License . SUPPLEMENTARY INFORMATION: http://www.psort.org/psortb/supplementaryinfo.html. Chem Biol, 2004 Oct, 11(10), 1403 - 11 Novel avilamycin derivatives with improved polarity generated by targeted gene disruption; Weitnauer G et al.; The oligosaccharide antibiotics avilamycin A and C are produced by Streptomyces viridochromogenes Tu57 . Both consist of a heptasaccharide chain, which is attached to a polyketide-derived dichloroisoeverninic acid moiety . They show excellent antibiotic activity against Gram-positive bacteria . Both molecules are modified by O-methylation at different positions, which contributes to poor water solubility and difficulties in galenical drug development . In order to generate novel avilamycin derivatives with improved polarity and improved pharmacokinetic properties, we generated a series of mutants with one, two, or three mutated methyltransferase genes . Based on the structure of the novel avilamycin derivatives, the exact function of three methyltransferases, AviG2, AviG5, and AviG6, involved in avilamycin biosynthesis could be assigned. J Biol Chem, 2004 Dec 24, 279(52), 54023 - 31 Epub 2004 Oct 13. The function of mitogen-activated protein kinase phosphatase-1 in peptidoglycan-stimulated macrophages; Shepherd EG et al.; Mitogen-activated protein (MAP) kinases play a pivotal role in the macrophages in the production of proinflammatory cytokines triggered by lipopolysaccharides . However, their function in the responses of macrophages to Gram-positive bacteria is poorly understood . Even less is known about the attenuation of MAP kinase signaling in macrophages exposed to Gram-positive bacteria . In the present study, we have investigated the regulation of MAP kinases and the role of MAP kinase phosphatase (MKP)-1 in the production of pro-inflammatory cytokines using murine RAW264.7 and primary peritoneal macrophages after peptidoglycan stimulation . Treatment of macrophages with peptidoglycan resulted in a transient activation of JNK, p38, and extracellular signal-regulated kinase . Most interestingly, MKP-1 expression was potently induced by peptidoglycan, and this induction was concurrent with MAP kinase dephosphorylation . Triptolide, a diterpenoid triepoxide, potently blocked the induction of MKP-1 by peptidoglycan and prolonged the activation of JNK and p38 . Overexpression of MKP-1 substantially attenuated the production of tumor necrosis factor (TNF)-alpha induced by peptidoglycan, whereas knockdown of MKP-1 by small interfering RNA substantially increased the production of both TNF-alpha and interleukin-1 beta . Finally, we found that in primary murine peritoneal macrophages, MKP-1 induction following peptidoglycan stimulation also coincided with inactivation of JNK and p38 . Blockade of MKP-1 induction resulted in a sustained activation of both JNK and p38 in primary macrophages . Our results reveal that MKP-1 critically regulates the expression of TNF-alpha and interleukin-1 beta in RAW264.7 cells and further suggest a central role for this phosphatase in controlling the inflammatory responses of primary macrophages to Gram-positive bacterial infection. Int J Lepr Other Mycobact Dis, 2004 Sep, 72(3), 327 - 30 False positive reaction of the immunohistochemistry technique using anti-BCG polyclonal antibodies to identify Mycobacterium leprae in wild nine-banded armadillos; Deps PD et al.; The authors studied 66 wild nine-banded armadillos from Brazil . The ear samples were collected and Ziehl-Neelsen or Fite-Faraco stains were performed, as well as immunostaining using polyclonal BCG antibody, to avaluate the presence of the Mycobacterium leprae . The AFB were not detected by the Ziehl-Neelsen or Fite-Faraco staining, neither immunoexpression of the BCG marker . However, many normal structures from the ears of the nine-banded armadillos, such as condrocytes, condroblasts, fibroblasts and endothelial cells, and Gram positive bacteria cocci, showed false positive reaction by the BCG marker . The authors discuss the use of the immunohistochemical studies with the polyclonal BCG antibody to identify M . leprae antigens in wild armadillos. Appl Microbiol Biotechnol, 2004 Dec, 66(2), 174 - 9 Epub 2004 Dec. A new ether bond-splitting enzyme found in Gram-positive polyethylene glycol 6000-utilizing bacterium, Pseudonocardia sp . strain K1; Yamashita M et al.; Pseudonocardia sp . strain K1 is the only Gram-positive bacterium among the bacteria aerobically metabolizing polyethylene glycol (PEG) . Generally, PEG is metabolized by an oxidative pathway in which a terminal alcohol group of PEG is oxidized to aldehyde and to carboxylic acid and then an ether bond is oxidatively cleaved . As the cell-free extract of Pseudonocardia sp . strain K1 has PEG dehydrogenase, PEG aldehyde dehydrogenase and diglycolic acid (DGA) dehydrogenase (DGADH) activities, all of which are constitutively formed, the strain has a metabolic pathway similar to that so far known . We purified an ether bond-splitting enzyme as DGADH . The molecular mass of the enzyme was estimated to be 55 kDa; and it consisted of two identical subunits . The enzyme oxidatively cleaved both an ether bond of PEG 3000 dicarboxylic acid and DGA . The N-terminal amino acid sequence of the purified enzyme had high homology with various superoxide dismutases and the enzyme had also superoxide dismutase activity . The atomic absorption spectrum showed that approximately one atom of Fe was included in each subunit of the enzyme . DGADH activity increased in the cells grown in a PEG medium supplemented with FeCl(3) . Thus, we concluded that the enzyme purified from Pseudonocardia sp . strain K1 is a new ether bond-splitting enzyme. J Clin Microbiol, 2004 Oct, 42(10), 4891 - 2 Rothia dentocariosa septicemia without endocarditis in a neonatal infant with meconium aspiration syndrome; Shin JH et al.; Rothia dentocariosa, a gram-positive coccoid- to rod-shaped bacterium with irregular morphology, is a rare cause of bacteremia in patients without endocarditis . We report the first case of R . dentocariosa septicemia without endocarditis, which occurred in a neonatal infant with meconium aspiration syndrome. J Med Assoc Thai, 2004 Aug, 87(8), 921 - 34 Pathology and etiology of 110 consecutively removed aortic valves; Chuangsuwanich T et al.; OBJECTIVES: To study the pathology and determine the etiology and prevalence of aortic valve disease from surgically removed aortic valve specimens . MATERIAL AND METHOD: All the native surgically excised aortic valves (AV) received from June 1997 to March 1999 (22 months) were studied macroscopically including cuspal measurements and microscopically . By preoperative echocardiographic and macroscopic studies, they were classified into functional disorders of predominant aortic stenosis (AS), aortic stenosis with regurgitation (AS-AR) and predominant aortic regurgitation (AR) . The patients' medical records were reviewed and the clinical information was extracted . The etiology was determined according to the macroscopic, microscopic and clinical findings . RESULTS: Among 110 AV (76 isolated AV and 34 with concomitant mitral valves from patients aged 15-96 years, mean age 47.54 years; male:female = 1.39:1) there were 25 AS (22.73%), 34 AS-AR (30.91%) and 51 AR (46.36%) cases . Eighty-four (76.36%) were tricuspid, 16 (14.54%) were bicuspid and 10 were undetermined . Cuspal measurements of each disease were provided and compared . All AS specimens were related to moderate to severe calcification and causes included postinflammatory disease (14 cases, 56%; age range 38-67 years, mean age 53.29 years, male:female = 0.56:1), degenerative calcific change (11 cases, 44%, age range 56-76 years, male:female = 1.2:1; mean age 69 years of 5 tricuspid AV and 60.83 years of 6 bicuspid AV) . In AS-AR, 29 cases (85.29%; mean age 47.10 years; male:female = 1.23:1) were attributable to postinflammatory disease and 5 cases (mean age 70.20 years; male:female = 1.5:1) to degenerative calcific change . In pure AR, there were 21 cases (age range 15-65 years, mean age 29.76 years) of postinflammatory disease, 14 cases of infective endocarditis (IE) and postIE (age range 20-63 years, mean age 42.21 years; all 10 IE cases contained gram positive cocci), 1 case (age 55 years) of bicuspid calcific change, 8 cases of AV with dilated valve ring, 5 cases of miscellaneous causes and 2 cases of indeterminate etiology . Aschoff bodies were found in 3 AR cases . Four of 18 postinflammatory AS-AR and 4 of 14 postinflammatory disease AR cases had past history of rheumatic fever . One postinflammatory AS also had infective endocarditis from gram positive cocci without clinical sign . Severe degenerative calcific change had a higher incidence of underlying diabetes (3 of 15 cases, 20%), hypertension (8 of 14 cases, 57.14%) and dyslipoproteinemia (9 of 13 cases, 69.23%) in comparison with 3.37% (3/89) for diabetes, 9.09% (8/88) for hypertension and 30.99% (22/71) for dyslipoproteinemia in other AV diseases in combination . CONCLUSION: The three common causes of severe AV functional disorders were postinflammatory disease (58.18%), degenerative calcific change (15.45%) and IE-postIE (12.72%) . Underlying diseases of severe degenerative calcific change included hypertension, dyslipoproteinemia and diabetes . Macroscopic and microscopic examinations together with clinical information, echocardiographic findings and operative details are important in evaluating the etiology of valvular diseases especially in severely calcified specimens. J Immunol, 2004 Oct 15, 173(8), 5229 - 37 Bacterial lipoprotein delays apoptosis in human neutrophils through inhibition of caspase-3 activity: regulatory roles for CD14 and TLR-2; Power CP et al.; The human sepsis syndrome resulting from bacterial infection continues to account for a significant proportion of hospital mortality . Neutralizing strategies aimed at individual bacterial wall products (such as LPS) have enjoyed limited success in this arena . Bacterial lipoprotein (BLP) is a major constituent of the wall of diverse bacterial forms and profoundly influences cellular function in vivo and in vitro, and has been implicated in the etiology of human sepsis . Delayed polymorphonuclear cell (PMN) apoptosis is a characteristic feature of human sepsis arising from Gram-negative or Gram-positive bacterial infection . Bacterial wall product ligation and subsequent receptor-mediated events upstream of caspase inhibition in neutrophils remain incompletely understood . BLP has been shown to exert its cellular effects primarily through TLR-2, and it is now widely accepted that lateral associations with the TLRs represent the means by which CD14 communicates intracellular messages . In this study, we demonstrate that BLP inhibits neutrophil mitochondrial membrane depolarization with a subsequent reduction in caspase-3 processing, ultimately leading to a significant delay in PMN apoptosis . Pretreatment of PMNs with an anti-TLR-2 mAb or anti-CD14 mAb prevented BLP from delaying PMN apoptosis to such a marked degree . Combination blockade using both mAbs completely prevented the effects of BLP (in 1 and 10 ng/ml concentrations) on PMN apoptosis . At higher concentrations of BLP, the antiapoptotic effects were observed, but were not as pronounced . Our findings therefore provide the first evidence of a crucial role for both CD14 and TLR-2 in delayed PMN apoptosis arising from bacterial infection. Surgery, 2004 Oct, 136(4), 937 - 43 Immunoglobulin A protease is a virulence factor for gram-negative pneumonia; Diebel LN et al.; BACKGROUND: Secretory immunoglobulin A (SIgA) is the principal immunologic defense of respiratory and other mucosal surfaces in the body . SIgA is relatively stable in mucosal secretions . However, cleavage of SIgA by bacterial proteases might render it immunologically inactive and thus contribute to the development of pneumonia as well as other infections . Bacterial species and infection sites might be important in the expression of bacterial protease activity and serve as the impetus to this study . METHODS: Bacterial isolates from respiratory and nonrespiratory sites were incubated with SIgA in vitro . SIgA degradation was determined by size exclusion ultrafiltration and gel electrophoresis . RESULTS: IgA protease activity was evident in gram-negative but not gram-positive respiratory isolates . Gram-negative isolates from nonrespiratory sources did not exhibit IgA protease activity . CONCLUSIONS: Expression of IgA protease activity might be important in the development and subsequent outcome of gram-negative pneumonia in the patient in the surgical intensive care unit. Ann Pharmacother, 2004 Nov, 38(11), 1855 - 9 Epub 2004 Oct 05. Probable vancomycin-induced neutropenia; Segarra-Newnham M et al.; OBJECTIVE: To report a case of vancomycin-induced neutropenia and provide a review of the literature . CASE SUMMARY: A 64-year-old white man was treated with intravenous vancomycin 1.5 g/day for finger osteomyelitis . He developed neutropenia after 21 days of vancomycin therapy . The absolute neutrophil count reached a nadir of 418 cells/mm(3) during vancomycin use and returned to normal 7 days after its discontinuation . The eosinophil count was also elevated during the neutropenic episode and probably related to vancomycin . Based on the Naranjo probability scale, the reaction was probably related to vancomycin use . DISCUSSION: Articles describing cases of vancomycin-induced neutropenia were identified . All patients developed neutropenia as a result of vancomycin therapy >/=12 days . Neutrophil counts generally increased following discontinuation of vancomycin . One article reported successful resolution of neutropenia and infection by switching the patient's therapy to the structurally related antibiotic agent teicoplanin . Other patients were continued on vancomycin therapy, and neutropenia was treated with moderate to good success with filgrastim . Rechallenge was not generally attempted . The mechanism of neutropenia caused by vancomycin is unclear, but appears to be immune-mediated . CONCLUSIONS: Vancomycin therapy should not be prolonged unless absolutely necessary, and therapy should be reserved for patients with clear indications for the drug, such as infections due to gram-positive organisms resistant to other therapies . Patients should have periodic assessment of white blood cell and neutrophil counts with consideration to discontinue vancomycin if neutropenia develops. J Bacteriol, 2004 Oct, 186(20), 6864 - 75 Isolation and characterization of ribosomes and translation initiation factors from the gram-positive soil bacterium Streptomyces lividans; Day JM et al.; A primer extension inhibition (toeprint) assay was developed using ribosomes and ribosomal subunits from Streptomyces lividans . This assay allowed the study of ribosome binding to streptomycete leaderless and leadered mRNA . Purified 30S subunits were unable to form a ternary complex on aph leaderless mRNA, whereas 70S ribosomes could form ternary complexes on this mRNA . 30S subunits formed ternary complexes on leadered aph and malE mRNA . The translation initiation factors (IF1, IF2, and IF3) from S . lividans were isolated and included in toeprint and filter binding assays with leadered and leaderless mRNA . Generally, the IFs reduced the toeprint signal on leadered mRNA; however, incubation of IF1 and IF2 with 30S subunits that had been washed under high-salt conditions promoted the formation of a ternary complex on aph leaderless mRNA . Our data suggest that, as reported for Escherichia coli, initiation complexes with leaderless mRNAs might use a novel pathway involving 70S ribosomes or 30S subunits bound by IF1 and IF2 but not IF3 . Some mRNA-ribosome-initiator tRNA reactions that yielded weak or no toeprint signals still formed complexes in filter binding assays, suggesting the occurrence of interactions that are not stable in the toeprint assay. Toxicol In Vitro, 2004 Dec, 18(6), 797 - 803 Proliferative responses observed following vancomycin treatment in renal proximal tubule epithelial cells; King DW et al.; Vancomycin (VAN) is a glycopeptide antibiotic used to treat gram-positive infections . Nephrotoxicity is a common side effect observed with vancomycin therapy . However, the mechanism of vancomycin-induced nephrotoxicity has not been fully characterized . In this study we examined the effect of vancomycin on cellular proliferation in renal proximal tubule cells . A dose- and time-dependent increase in cell number and total cellular protein was observed following vancomycin exposure . Vancomycin exposure also caused an increase in BrdU incorporation followed by the accumulation of renal proximal tubule cells in G(2)/M phase of the cell cycle . These effects were inhibited by pretreatment with the mitogen-activated protein kinase inhibitor, PD098059, suggesting an association between the cell proliferative effect of VAN and the induction of the mitogen-activated protein kinase signaling pathway . Mitochondrial function in renal proximal tubule cells was assessed using oxygen consumption and ATP concentrations . We observed an increase in oxygen consumption and ATP concentrations following short-term exposure to vancomycin . Together, our data suggest that vancomycin treatment produces alterations in mitochondrial function that coincide with a cell proliferative response in renal proximal tubule epithelial cells. J Heart Lung Transplant, 2004 Sep, 23(9), 1119 - 22 Sideroblastic anemia due to linezolid in a patient with a left ventricular assist device; Montpetit MC et al.; An acquired form of sideroblastic anemia has been described in association with several drugs, especially anti-microbial agents . A case of sideroblastic anemia is presented in a patient with a left ventricular assist device drive-line infection who was receiving linezolid, an antibiotic used for serious infections with gram-positive organisms . This patient's anemia resolved after discontinuation of the drug; he subsequently underwent an uncomplicated orthotopic heart transplant with no recurrence of anemia . As linezolid has been shown to have hematologic side effects, blood count monitoring is recommended in patients receiving this drug for long-term therapy. Curr Opin Microbiol, 2004 Oct, 7(5), 439 - 44 Microbiology and drug resistance mechanisms of fully resistant pathogens; Walsh FM et al.; The acquisition of vancomycin resistance by Gram-positive bacteria and carbapenem resistance by Gram-negative bacteria has rendered some hospital-acquired pathogens impossible to treat . The resistance mechanisms employed are sophisticated and very difficult to overcome . Unless alternative treatment regimes are initiated soon, our inability to treat totally resistant bacteria will halt other developments in medicine . In the community, Gram-positive bacteria responsible for pneumonia could become totally resistant leading to increased mortality from this common infection, which would have a more immediate impact on our current lifestyles. Nat Immunol, 2004 Nov, 5(11), 1175 - 80 Epub 2004 Sep 26. Function of the drosophila pattern-recognition receptor PGRP-SD in the detection of Gram-positive bacteria; Bischoff V et al.; The activation of an immune response requires recognition of microorganisms by host receptors . In drosophila, detection of Gram-positive bacteria is mediated by cooperation between the peptidoglycan-recognition protein-SA (PGRP-SA) and Gram-negative binding protein 1 (GNBP1) proteins . Here we show that some Gram-positive bacterial species activate an immune response in a PGRP-SA- and GNBP1-independent manner, indicating that alternative receptors exist . Consistent with this, we noted that PGRP-SD mutants were susceptible to some Gram-positive bacteria and that a loss-of-function mutation in PGRP-SD severely exacerbated the PGRP-SA and GNBP1 mutant phenotypes . These data indicate that PGRP-SD can function as a receptor for Gram-positive bacteria and shows partial redundancy with the PGRP-SA-GNBP1 complex. Int J Syst Evol Microbiol, 2004 Sep, 54(Pt 5), 1557 - 60 Helcococcus sueciensis sp . nov., isolated from a human wound; Collins MD et al.; A previously undescribed, Gram-positive, catalase-negative, coccus-shaped organism that originated from a human wound was subjected to taxonomic study . On the basis of its cellular morphology and the results of biochemical testing, the unknown organism was identified tentatively as a member of the genus Helcococcus, but it did not correspond to either of the two recognized species of this genus . Comparative 16S rRNA gene sequencing studies confirmed that the bacterium was associated phylogenetically with the genus Helcococcus, with the unidentified organism forming a hitherto unknown subline within the genus . On the basis of biochemical, molecular chemical and molecular phylogenetic evidence, it is proposed that the unknown organism that was recovered from a human wound should be classified as a novel species of the genus Helcococcus, namely Helcococcus sueciensis sp . nov . The type strain is CCUG 47334T (= CIP 108183T). J Nat Prod, 2004 Sep, 67(9), 1476 - 82 Antineoplastic agents . 529 . Isolation and structure of nootkastatins 1 and 2 from the Alaskan yellow cedar Chamaecyparis nootkatensis; Pettit GR et al.; The yellow cedar tree, Chamaecyparis nootkatensis, collected in southeast Alaska was evaluated as a potential source of new anticancer agents . Two new diterpene anticancer constituents termed nootkastatins 1 (4) and 2 (5) were isolated along with three previously known diterpene cancer cell growth inhibitors where two were reported as synthetic modifications of totarol and not previously found in nature . All five diterpene structures were established by HRMS and 1D and 2D NMR spectroscopic analyses combined with three X-ray crystal structure determinations (2, 3, and 5) . Against a panel of six human cancer cell lines, this series of diterpenes exhibited inhibition over the range GI(50) 0.75-2.0 microg/mL, and all inhibited the growth of Gram-positive bacteria and fungi. Infect Immun, 2004 Oct, 72(10), 5704 - 11 The phosphatidylinositol 3-kinase/protein kinase B signaling pathway is activated by lipoteichoic acid and plays a role in Kupffer cell production of interleukin-6 (IL-6) and IL-10; Dahle MK et al.; Sepsis caused by gram-positive bacteria lacking lipopolysaccharide (LPS) has become a major and increasing cause of mortality in intensive-care units . We have recently demonstrated that the gram-positive-specific bacterial cell wall component lipoteichoic acid (LTA) stimulates the release of the proinflammatory cytokines in Kupffer cells in culture . In the present study, we have started to assess the signal transduction events by which LTA induces the production of tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and the anti-inflammatory cytokine IL-10 in rat Kupffer cells . LTA was found to trigger phosphorylation of mitogen-activated protein kinases (MAPK) (p38 MAPK and ERK 1/2) and protein kinase B (PKB) . Compared to LPS, LTA was more potent in inducing PKB phosphorylation after 40 min, although we found that the cytokine responses were similar . For both bacterial molecules, blocking phosphatidylinositol 3-kinase (PI3-K; Ly294002) or Janus kinase 2 (JAK-2; AG490) particularly affected the induction of IL-6 and IL-10 release, whereas TNF-alpha levels were strongly reduced by inhibition of Src family tyrosine kinases (PP2) . All three cytokines were reduced by inhibition of p38 MAPK (SB202190) or the broad-range tyrosine kinase inhibitor genistein, whereas IL-6 release was particularly blocked by inhibition of ERK 1/2 (PD98059) . Divergences in the regulatory pathways controlling TNF-alpha, IL-10, and IL-6 production in Kupffer cells following LPS or LTA stimulation may create a basis for understanding how the balance between pro- and anti-inflammatory cytokines is regulated in the liver following infections by gram-positive or gram-negative bacteria. Phytochemistry, 2004 Aug, 65(16), 2369 - 72 Iridoids from the aerial parts of Verbena littoralis (Verbenaceae); Castro-Gamboa I et al.; The iridoids, 6S-hydroxy-8S-methyl-4-methylene-hexahydro-cyclopenta{c}pyran-3-one and 6S,9S-dihydroxy-8S-methyl-4-methylene-hexahydro-cyclopenta{c}pyran-3-one, were isolated from the aerial parts of Verbena littoralis . Their structures and stereochemistry were elucidated by means of NMR spectral data analysis . Both compounds showed moderate in vitro activity against gram positive and negative bacteria as well as moderate in vivo intestinal peristaltic action in mouse . The iridoids also showed moderate free radical scavenging activity against l,l-diphenyl-2-picrylhydrazyl (DPPH) as well as antioxidant activity, the latter being evidenced by redox properties measured using E1CD-HPLC. Immunology, 2004 Oct, 113(2), 224 - 33 Histamine induces Toll-like receptor 2 and 4 expression in endothelial cells and enhances sensitivity to Gram-positive and Gram-negative bacterial cell wall components; Talreja J et al.; Histamine is a major inflammatory molecule released from the mast cell, and is known to activate endothelial cells . However, its ability to modulate endothelial responses to bacterial products has not been evaluated . In this study we determined the ability of histamine to modulate inflammatory responses of endothelial cells to Gram-negative and Gram-positive bacterial cell wall components and assessed the role of Toll-like receptors (TLR) 2 and 4 in the co-operation between histamine and bacterial pathogens . Human umbilical vein endothelial cells (HUVEC) were incubated with lipopolysaccharide (LPS), lipoteichoic acid (LTA), or peptidoglycan (PGN) in the presence or absence of histamine, and the expression and release of interleukin-6 (IL-6), and NF-kappaB translocation were determined . The effect of histamine on the expression of mRNA and proteins for TLR2 and TLR4 was also evaluated . Incubation of HUVEC with LPS, LTA and PGN resulted in marked enhancement of IL-6 mRNA expression and IL-6 secretion . Histamine alone markedly enhanced IL-6 mRNA expression in HUVEC, but it did not stimulate proportional IL-6 release . When HUVEC were incubated with LPS, LTA, or PGN in the presence of histamine marked amplification of both IL-6 production and mRNA expression was noted . HUVEC constitutively expressed TLR2 and TLR4 mRNA and proteins, and these were further enhanced by histamine . The expression of mRNAs encoding MD-2 and MyD88, the accessory molecules associated with TLR signalling, were unchanged by histamine treatment . These results demonstrate that histamine up-regulates the expression of TLR2 and TLR4 and amplifies endothelial cell inflammatory responses to Gram-negative and Gram-positive bacterial components. Curr Drug Targets Infect Disord, 2004 Sep, 4(3), 207 - 15 Distribution of macrolide, lincosamide, streptogramin, ketolide and oxazolidinone (MLSKO) resistance genes in Gram-negative bacteria; Roberts MC; A number of different mechanisms of macrolide resistance have been described in Gram-negative bacteria . These include 16 acquired genes (esterases, phosphorylases, rRNA methylases, and effluxes) and include those thought to be unique to Gram-negative bacteria (both esterases and two of the phosphorylases) and those shared with Gram-positive bacteria (one phosphorylase) and those primarily of Gram-positive origin (rRNA methylases and efflux genes) . In addition, mutations, which modify the 23S rRNA, ribosomal proteins L4 and/or L22, and/or changes in expression of innate efflux systems which occur by missense, deletion and/or insertion events have been described in five Gram-negative groups, while an innate transferase conferring resistance to streptogramin A has been identified in a sixth genus . However, the amount of information on both acquisition and mutations leading to macrolide, lincosamides, streptogramins, ketolides and oxazolidinones (MLSKO) resistance is limited . As a consequence this review likely underestimates the true distribution of acquired genes and mutations in Gram-negative bacteria . As use of these drugs increases, it is likely that interaction between members of the MLSKO antibiotic family and Gram-negative bacteria will continue to change resistance to these antibiotics; by mutations of existing genes as well as by acquisition and perhaps mutations of acquired resistant genes in these organisms and more work needs to be done to get a clearer picture of what is in the Gram-negative population now, such that changes can be monitored. Proteomics, 2004 Oct, 4(10), 3068 - 77 Highly phosphorylated bacterial proteins; Rosen R et al.; We show in Gram-negative and Gram-positive bacteria the appearance of highly acidic proteins, which are highly phosphorylated . This group of proteins includes many cellular proteins, such as chaperones, biosynthetic, and metabolic enzymes . These proteins accumulate under stress conditions or under conditions, which overload the proteolytic system . Pulse chase experiments using radioactive phosphate indicate that the phosphorylated proteins have a short half-life, suggesting that they could be degradation intermediates . Moreover, results from in vitro experiments in Escherichia coli indicated that ribosomal proteins become susceptible to proteolysis after polyphosphorylation . Therefore, it is possible that the highly phosphorylated proteins represent a group of proteins tagged for degradation by phosphorylation . Such a tagging process may be involved in a general bacterial degradation pathway. J Biol Chem, 2004 Nov 26, 279(48), 50591 - 600 Epub 2004 Nov 26. Crystal structure of bacterial inorganic polyphosphate/ATP-glucomannokinase . Insights into kinase evolution; Mukai T et al.; Inorganic polyphosphate (poly(P)) is a biological high energy compound presumed to be an ancient energy carrier preceding ATP . Several poly(P)-dependent kinases that use poly(P) as a phosphoryl donor are known to function in bacteria, but crystal structures of these kinases have not been solved . Here we present the crystal structure of bacterial poly(P)/ATP-glucomannokinase, belonging to Gram-positive bacterial glucokinase, complexed with 1 glucose molecule and 2 phosphate molecules at 1.8 A resolution, being the first among poly(P)-dependent kinases and bacterial glucokinases . The poly(P)/ATP-glucomannokinase structure enabled us to understand the structural relationship of bacterial glucokinase to eucaryotic hexokinase and ADP-glucokinase, which has remained a matter of debate . These comparisons also enabled us to propose putative binding sites for phosphoryl groups for ATP and especially for poly(P) and to obtain insights into the evolution of kinase, particularly from primordial poly(P)-specific to ubiquitous ATP-specific proteins. Eur J Pharmacol, 2004 Sep 13, 498(1-3), 295 - 301 15d-prostaglandin J2 reduces multiple organ failure caused by wall-fragment of Gram-positive and Gram-negative bacteria; Dugo L et al.; Septic shock is still the major cause of death in surgical intensive care units . Both gram-positive (G+) and gram-negative (G-) bacteria have been isolated in the blood of a large portion of septic patients, and these polymicrobial infections often have a higher mortality than infections due to a single organism . Cell wall fragments from G+ and G- bacteria synergise to cause shock and multiple organ dysfunction in vivo (G+/G- shock) . Male Wistar rats were anaesthetised and received a coadministration of wall fragments from G+ and G- bacteria, Staphilococcus aureus (S . aureus) peptidoglycan {0.3 mg/kg, intravenously (i.v.)} and Escherichia coli (E . coli) lipopolysaccharide (1 mg/kg, i.v.) or vehicle (saline, 1 ml/kg, i.v.) . G+/G- shock for 6 h resulted in an increase in serum levels of creatinine (indicator of renal dysfunction), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (gamma-GT), bilirubin (markers for hepatic injury and dysfunction) and creatine kinase (CK, an indicator of neuromuscular, skeletal muscle or cardiac injury) . Pretreatment of rats with the peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonist 15d-prostaglandin J2 (0.3 mg/kg, i.v., 30 min prior to G+/G-) reduced the multiple organ injury/dysfunction caused by coadministration of peptidoglycan+lipopolysaccharide . The selective PPAR-gamma antagonist GW9662 (2-Chloro-5-nitrobenzanilide) (1 mg/kg, i.v., given 45 min prior to G+/G-) abolished the protective effects of 15d-prostaglandin J2 . 15d- prostaglandin J2 did not affect the biphasic fall in blood pressure or the increase in heart rate caused by administration of peptidoglycan+lipopolysaccharide . The mechanism(s) of the protective effect of this cyclopentenone prostaglandin are-at least in part-PPAR-gamma dependent, as the protection afforded by 15d-prostaglandin J2 was reduced by the PPAR-gamma antagonist GW9662 . We propose that 15d-prostaglandin J2 or other ligands for PPAR-gamma may be useful in the therapy of the organ injury associated with septic shock. Trends Genet, 2004 Oct, 20(10), 475 - 9 Conserved regulatory motifs in bacteria: riboswitches and beyond; Abreu-Goodger C et al.; We present a computational approach that identifies regulatory elements conserved across phylogenetically distant organisms . Intergenic regulatory regions were clustered by orthology of the adjacent genes, and an iterative process was applied to search for significant motifs, enabling new elements of the putative regulon to be added in each cycle . With this approach, we identified highly conserved riboswitches and the Gram positive T-box . Interestingly, we identified many other regulatory systems that appear to depend on conserved RNA structures. Carbohydr Res, 2004 Aug 23, 339(12), 2111 - 5 Crystal and molecular structure and absolute configuration of lincomycin hydrochloride monohydrate; Rajeswaran M et al.; Lincomycin is a broad-spectrum antibiotic synthesized by Streptomyces lincolnensis that is particularly active against Gram-positive bacteria . It is widely used in human and veterinary applications . The crystal structure of lincomycin has been undertaken with a view to obtain the conformational and structural features of the drug in order to afford a comparison of its structural features with other aminoglycoside antibiotics . We report here the details of its structural and conformational features as determined by single-crystal X-ray crystallography . Crystals of lincomycin hydrochloride are orthorhombic, space group P2(1)2(1)2, with the cell dimensions a=18.5294(3) Angstroms, b=20.5980(4) Angstroms, c=6.17380(10) Angstroms, V=2356.35(7) Angstroms3 . The structure was solved using X-ray diffraction data and refined to a final R-value of 0.0391 for 2321 reflections (I > or = 2sigma) . The absolute configuration was established using the anomalous dispersion of the sulfur and chlorine atoms in the structure . The molecule consists of an amino acid linked by an amide group to a monosaccharide of galactose stereochemistry . A network of hydrogen-bonds stabilizes the crystal structure. Microbiology, 2004 Sep, 150(Pt 9), 2795 - 806 Sensing and responding to diverse extracellular signals? Analysis of the sensor kinases and response regulators of Streptomyces coelicolor A3(2); Hutchings MI et al.; Streptomyces coelicolor is a Gram-positive soil bacterium that undergoes a complex developmental life cycle . The genome sequence of this organism was recently completed and has revealed the presence of over 60 sigma factors and a multitude of other transcriptional regulators, with a significant number of these being putative two-component signal transduction proteins . The authors have used the criteria established by Hoch and co-workers (Fabret et al., 1999, J Bacteriol 181, 1975-1983) to identify sensor kinase and response regulator genes encoded within the S . coelicolor genome . This analysis has revealed the presence of 84 sensor kinase genes, 67 of which lie adjacent to genes encoding response regulators . This strongly suggests that these paired genes encode two-component systems . In addition there are 13 orphan response regulators encoded in the genome, several of which have already been characterized and are implicated in development and antibiotic production, and 17 unpaired and as yet uncharacterized sensor kinases . This article attempts to infer useful information from sequence analysis and reviews what is currently known about the two-component systems, unpaired sensor kinases and orphan response regulators of S . coelicolor from both published reports and the authors' own unpublished data. Appl Environ Microbiol, 2004 Sep, 70(9), 5057 - 65 Comparison of diversities and compositions of bacterial populations inhabiting natural forest soils; Hackl E et al.; The diversity and composition of soil bacterial communities were compared among six Austrian natural forests, including oak-hornbeam, spruce-fir-beech, and Austrian pine forests, using terminal restriction fragment length polymorphism (T-RFLP, or TRF) analysis and sequence analysis of 16S rRNA genes . The forests studied differ greatly in soil chemical characteristics, microbial biomass, and nutrient turnover rates . The aim of this study was to relate these differences to the composition of the bacterial communities inhabiting the individual forest soils . Both TRF profiling and clone sequence analysis revealed that the bacterial communities in soils under Austrian pine forests, representing azonal forest types, were distinct from those in soils under zonal oak-hornbeam and spruce-fir-beech forests, which were more similar in community composition . Clones derived from an Austrian pine forest soil were mostly affiliated with high-G+C gram-positive bacteria (49%), followed by members of the alpha-Proteobacteria (20%) and the Holophaga/Acidobacterium group (12%) . Clones in libraries from oak-hornbeam and spruce-fir-beech forest soils were mainly related to the Holophaga/Acidobacterium group (28 and 35%), followed by members of the Verrucomicrobia (24%) and the alpha-Proteobacteria (27%), respectively . The soil bacterial communities in forests with distinct vegetational and soil chemical properties appeared to be well differentiated based on 16S rRNA gene phylogeny . In particular, the outstanding position of the Austrian pine forests, which are determined by specific soil conditions, was reflected in the bacterial community composition. J Med Chem, 2004 Sep 9, 47(19), 4693 - 709 A prodrug approach toward the development of water soluble fluoroquinolones and structure--activity relationships of quinoline-3-carboxylic acids; B