J Infect Dis, 2005 Feb 15, 191(4), 579 - 87 Epub 2005 Jan 14. Host Adaptation and Immune Modulation Are Mediated by Homologous Recombination in Helicobacter pylori; Robinson K et al.; Rearrangement of genomic DNA via homologous recombination provides an alternative mechanism of gene regulation that is essential for successful colonization of the gastric mucosa by Helicobacter pylori . Inoculation of outbred mice with the H . pylori SS1 wild-type strain elicited a T helper (Th) 2 response and established a persistent infection . In contrast, inoculation with an isogenic H . pylori strain defective for homologous recombination elicited a Th1-mediated immune response and clearance of infection within 70 days . We, therefore, demonstrate that recombination is critical for mediating persistence of a microbial pathogen through the induction of ineffective immune responses. Anticancer Drugs, 2005 Feb, 16(2), 145 - 50 Cytotoxicities of three rebeccamycin derivatives in the National Cancer Institute screening of 60 human tumor cell lines; Moreau P et al.; Among the biologically active indolocarbazoles, rebeccamycin, a microbial metabolite produced by Saccharothrix aerocolonigenes, is a well-known topoisomerase (Topo) I poison . In the course of structure-activity relationship studies on rebeccamycin analogs, we have prepared a large number of indolocarbazole derivatives and have shown that, depending on the structural modifications, the cytotoxic effects may be, or not, directly correlated to DNA binding and Topo I inhibition . This suggests that if DNA binding and Topo I play a part in the biological activity of these compounds, other cellular targets might be involved . This paper reports the results of the antiproliferative activities (evaluated in the National Cancer Institute's in vitro panel of 60 tumor cell lines) and the results of a COMPARE analysis run with rebeccamycin derivatives to identify other potential biological targets for these compounds. Blood Purif, 2004, 22 Suppl 2, 49 - 54 Selection of Dilutional Method for On-Line HDF, Pre- or Post-Dilution; Masakane I; Post- and pre-dilution methods equally have advantages and disadvantages, therefore we should choose the more favorable one for clinical use . However, it still remains controversial which technique is better in on-line HDF . In post-dilution, the clearances of small molecular uremic toxins increase as well as low molecular proteins, however the risk of albumin leakage caused by high transmembranous pressure (TMP) also increases . We can avoid the risk of albumin leakage because there are several maneuvers which aid in avoiding an increase of TMP, such as glucose infusion and gradual control of Q(f) . On the other hand, the pre-dilution method is rather safe in the risk of albumin leakage but has an obvious clearance loss of small molecular substances caused by a decreased dialysate flow rate . The influence of microbial contamination and acetate are rather severe in pre-dilution on-line HDF . These lines of evidence suggest that post-dilution on-line HDF is the best choice for treating chronic HD patients . Copyright (c) 2004 S . Karger AG, Basel. Clin Exp Immunol, 2005 Feb, 139(2), 314 - 22 Impaired interleukin (IL)-4-associated generation of CCR4-expressing T cells in neonates with hereditary allergy risk; Haddeland U et al.; Summary Reduced microbial exposure in early life may contribute to the increase of atopic diseases in 'westernized' societies but the underlying mechanisms remain elusive . The objective of this study was to examine how exposure to bacterial lipopolysaccharide (LPS) during early antigen encounter might influence the maturation of neonatal lymphoid cells, and to define possible differences in this respect between neonates with high risk of allergy due to a family history (FH(+)) and controls with no apparent hereditary risk (FH(-)) . Cord blood mononuclear cells from the FH(+) or FH(-) group were stimulated with pure LPS or beta-lactoglobulin (beta-LG) in the presence of LPS . T cell expression of chemokine receptors CCR4 and CXCR3 was determined by flow cytometry and reverse transcription-polymerase chain reaction (RT-PCR) . Cellular expression of interleukin (IL)-4 was analysed by quantitative RT-PCR, whereas interferon (IFN)-gamma was analysed by both quantitative RT-PCR and immunoassay . Stimulation with LPS, or beta-LG together with LPS, induced up-regulation of CCR4 (P < 0.05) and CXCR3 (P < 0.05) . For CCR4, such up-regulation was related to the level of IL-4 produced by the same T cells (r(S) = 0.49, P = 0.03), while CXCR3 expression was negatively correlated with the IL-4 levels (r(S) = -0.56, P = 0.02) . Compared with the FH(-) group, the FH(+) group showed a significantly lower capacity for generation of CCR4(+) T cells (mean percentage of total T cells: FH(+), 2.42%versus FH(-), 5.74%; P < 0.01), whereas induction of CXCR3 and IFN-gamma did not differ significantly between the two groups . When the immune system in early life encounters antigen together with LPS, the T cell potential for compartmentalized interaction with other immune cells might be increased by elevated CCR4- and CXCR3-expression levels . In neonates at hereditary allergy risk, this putative homeostatic mechanism could theoretically be jeopardized due to decreased up-regulation of CCR4 . Conversely, Th1 responses to antigen in the presence of LPS did not appear to be reduced compared with controls. Inflamm Res, 2004 Dec 1, 53(12), 644 - 52 Neutrophil recruitment in mast cell-dependent inflammation: inhibitory mechanisms of glucocorticoids; Schramm R et al.; Mast cells are strategically localized along the microvasculature in tissues in close contact with the external environment, such as the skin, lung and intestines . By releasing a multi-faceted spectrum of proinflammatory mediators, such as cytokines and chemokines, mast cells have the capacity to coordinate trafficking of leukocytes . Mast cells play a pathophysiological role in numerous inflammatory diseases as diverse as hypersensitivity reactions, ischemia/reperfusion injury and rheumatoid arthritis . On the other hand, mast cells act also as tissue sentinels and are critically involved in the host defensive response against microbial infection by stimulating neutrophil recruitment . Glucocorticoids are powerful agents frequently used in mast cell-dependent diseases, although the anti-inflammatory mechanisms of these compounds are not completely understood at present . In order to circumvent steroid-associated side-effects and develop more specific therapeutics, numerous studies have examined the mechanisms underlying glucocorticoid inhibition of mast cell-dependent neutrophil recruitment . Based on recent findings, it may be suggested that glucocorticoids selectively inhibit the expression and function of certain adhesion molecules and chemokines . This review summarizes current insights into the underlying mechanisms of mast cell-regulated tissue accumulation of neutrophils and the inhibitory effects of glucocorticoids. J Dairy Sci, 2005 Feb, 88(2), 466 - 9 Inactivation of penicillin g in milk using hydrogen peroxide; Hanway WH et al.; Milk antibiotic residues have been a public concern in recent years . The Grade A Pasteurized Milk Ordinance mandates that raw Grade A milk will test negative for beta-lactam antibiotic residues before processing . The purpose of this research was to investigate the ability of various levels of peroxide and heat to inactivate penicillin G in raw milk . Whole milk spiked to a mean of 436 +/- 15.1 (standard error of the mean) ppb of potassium penicillin G was treated with hydrogen peroxide at levels of 0.0, 0.09, 0.17, and 0.34% . Samples at each peroxide level (n = 6 per treatment) were treated as follows: 1) incubated at 54.4 degrees C for 3 h, 2) pasteurized at 62.8 degrees C for 30 min, 3) incubated and pasteurized as in treatments 1 and 2, or 4) received no further treatment . A beta-lactam competitive microbial receptor assay was used for quantification of penicillin G . Concentrations of penicillin in selected samples were determined by HPLC for a comparison of test methods . Treatments were evaluated relative to their ability to reduce milk penicillin G levels to below the safe level of 5 ppb.The 0.09% hydrogen peroxide level was ineffective for all treatments . Hydrogen peroxide at 0.17% lowered the mean penicillin G (+/- SEM) from 436 +/- 15.1 to 6 +/- 1.49 ppb using the incubated and pasteurized heat treatment . The 0.34% concentration of hydrogen peroxide was the most effective, inactivating penicillin G to a level well below the safe level of 5 ppb with the pasteurized heat treatment, with or without incubation. Biol Chem, 2004 Dec, 385(12), 1165 - 70 Conserved asparagine residue 54 of alpha-sarcin plays a role in protein stability and enzyme activity; Siemer A et al.; Asparagine 54 of alpha-sarcin is a conserved residue within the proteins of the ribotoxin family of microbial ribonucleases . It is located in loop 2 of the protein, which lacks repetitive secondary structure elements but exhibits a well-defined conformation . Five mutant variants at this residue have been produced and characterized . The spectroscopic characterization of these proteins indicates that the overall conformation is not changed upon mutation . Activity and denaturation assays show that Asn-54 largely contributes to protein stability, and its presence is a requirement for the highly specific inhibitory activity of these ribotoxins on ribosomes. Curr Opin Immunol, 2005 Feb, 17(1), 25 - 8 Manipulation of innate immunity by bacterial pathogens; Portnoy DA; The past decade has witnessed tremendous growth in two related fields: innate immunity and microbial pathogenesis . Many pathogens have evolved mechanisms to infect their hosts in the face of a fully functional innate immune system, and there are numerous examples by which pathogens avoid recognition and/or suppress inflammation . In this review, I suggest that pathogens not only survive the innate immune response, but use it to promote their pathogenesis. Biochem Pharmacol, 2005 Feb 1, 69(3), 365 - 71 Modulating the cytokine response to treat Helicobacter gastritis; Zavros Y et al.; The conventional view of gastric acid secretion is that a negative feedback mechanism arises in response to high acidity, such that somatostatin keeps G-cells and parietal cells from producing more gastrin and acid, respectively . When the stomach becomes infected, for example with Helicobacter pylori (H . pylori), the feedback mechanism is impaired . In animal models, our laboratory has demonstrated that other types of bacteria besides H . pylori can cause gastritis . For example, under conditions of low acidity, gastritis is secondary to bacterial overgrowth, not production of excessive acid, thus suggesting a new paradigm for the regulation of gastric acid secretion under inflammatory conditions . Cytokines, released during the gastric inflammatory response, including IFNgamma, TNFalpha and IL-1beta stimulate the G-cell to produce gastrin . Gastrin in turn triggers the release of acid, and hypergastrinemia suppresses somatostatin, the inhibitor of acid . The overall response results in maximal gastric acid output that acts as the stomach's most important anti-microbial agent . The increased acid secretion by the stomach in the presence of H . pylori seems to be part of the innate immune response, in that gastrin and somatostatin are reciprocally regulated by Th1 or Th2 cytokines, respectively . In a mouse model, we showed that octreotide, a somatostatin, analog, is an efficacious treatment for Helicobacter gastritis . In humans, octreotide might accelerate recovery from H . pylori infection, reducing the duration of antibiotic therapy. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz, 2005 Jan, 48(1), 43 - 53 {Comparison of methods for quantification of MVOC in indoor environments.}; Fischer G et al.; For several years now, MVOC have been regarded as indicators for microbial growth in indoor environments . Until now, a direct correlation between the occurrence of microfungi and MVOC could not be shown in scientific investigations . One reason may be that different analytical methods were applied, and moreover they were not validated sufficiently . The present investigation aimed to test the comparability of both methods (Tenax adsorption/thermal desorption; charcoal adsorption/elution) . It turned out that with both methods comparable results can be achieved if the technical handling of the calibration is standardized to a wide extent . Thus, highest demands have to be made on quality assurance . Therefore, it is necessary to formulate technical regulations for the quantification of MVOC. J Microbiol Methods, 2005 Mar, 60(3), 343 - 52 Increased precision of microbial RNA quantification using NASBA with an internal control; Patterson SS et al.; Detection and quantification of low abundance target RNA has wide utility in the fields of clinical diagnostics, environmental monitoring, gene expression analysis, and biodefense . Nucleic acid based sequence amplification (NASBA) is an isothermal amplification method that provides the sensitivity needed for these applications . However, the requirement for three separate enzymes in NASBA often results in a greater variability between replicate samples than that seen in PCR-based assays . To overcome this problem, we have adapted the bioMerieux Nuclisens Basic Kit and Nuclisens EasyQ Analyzer along with the introduction of a synthetic internal control RNA (IC-RNA) for quantification of potentially any RNA sequence . Using the rbcL gene from the Florida red tide organism Karenia brevis as our target, we describe a simple method to accurately quantify the native target by computing the ratio of the time to positivity (TTP) values for both the wild-type and IC-RNA, and plotting this ratio against the starting number of target molecules or cells . By utilizing this simple method, we have significantly increased our accuracy and precision of prediction over the standard TTP calculations. Dis Aquat Organ, 2004 Nov 23, 62(1-2), 1 - 16 Parasitism in species of Bathymodiolus (Bivalvia: Mytilidae) mussels from deep-sea seep and hydrothermal vents; Ward ME et al.; Bivalve species, especially mussels, are biomass dominants in many deep-sea chemosynthetic ecosystems . As in shallow-water environments, parasites are likely to be important factors in the population dynamics of bivalve communities in chemosynthetic ecosystems, but there has been little study of parasitism in deep-sea seep or vent molluscs . In this study, parasite types, diversity, prevalence, infection density and non-infectious indicators of stress or disease as related to host age, reproductive condition, and endosymbiont density were assessed in mussels (Bathymodiolus heckerae) from 2 seep sites and mussels (B . puteoserpentis) from 2 vent sites . We identified 10 microbial or parasitic agents in histological sections . Parasite types included 3 viral-like gut inclusions, 2 rickettsia-like gill inclusions, a rickettsia-like mantle inclusion, a bacterial gill-rosette, a chlamydia-like gut inclusion, gill-dwelling ciliates, and an unidentified inclusion in gut tissues . Parasite species richness was greater in seep mussels than in vent mussels, with the seep mussels possessing 9 types of parasites compared to 2 in the vent mussels . One of the viral-like inclusions infecting the seep mussel B . heckerae was pathogenic, causing lysis of the digestive tubules . The prevalence and intensity of infection by this pathogen were greater in hosts with shell lengths less than 100 mm . Mussels from all 4 sites also exhibited intense infiltration of tissues and blood spaces by enlarged hemocytes . Hemocytic infiltration (hemocytosis) showed variable degrees of severity that were not associated with other host factors examined. J Environ Qual, 2005 Jan-Feb, 34(1), 390 - 6 Effects of sedimentation on soil nutrient dynamics in riparian forests; Lockaby BG et al.; The influence of sedimentation rates on biogeochemistry of riparian forests was studied near ephemeral streams at Fort Benning, GA . Upper reaches of seven ephemeral streams had received varying rates of sedimentation stemming from erosion along unpaved roadways at the military installation . Two reference catchments were also included in the study . Decomposition of foliar litter, microbial C and N, N mineralization, and arthropod populations were compared within and among catchments . Rates of sedimentation over the past 25 yr ranged from 0 in references to 4.0 cm yr(-1) . Decomposition rates declined exponentially with sedimentation rates as low as 0.20 to 0.32 cm yr(-1) and appeared to reach an equilibrium at a sedimentation rate of 0.5 cm yr(-1) . Nitrogen mineralization and microbial C and N followed the same trend . Sedimentation had no discernible effect on arthropod populations . These data suggest that biogeochemical cycles may be altered by sedimentation rates that commonly occur in some floodplain forests. J Environ Qual, 2005 Jan-Feb, 34(1), 139 - 48 Ecosystem function in alluvial tailings after biosolids and lime addition; Brown S et al.; Municipal biosolids and agricultural limestone were incorporated into the surface of alluvial highly acidic, metal-contaminated mine tailings in Leadville, CO, in 1998 . Amended sites were seeded and a plant cover was subsequently established . A range of chemical and biological parameters were measured over time to determine if treatment was sufficient to restore ecosystem function . An uncontaminated upstream control (UUC), a contaminated vegetated area (CVA), and soils collected from the tailings deposits before amendment addition were used for comparison . Standard soil extracts showed decreases in extractable Pb, Zn, and Cd in the amended soils . Increased CO(2) evolution, reduced N(2)O, and elevated NO(3)(-) in the amended tailings indicated an active microbial community . Levels of CO(2) and NO(3)(-) were elevated in comparison with the CVA and UUC . Ryegrass (Lolium perenne L.) and earthworm (Eisenia foetida) survival and metal uptake values were similar in amended tailings to a laboratory control soil . Ryegrass and worms in unamended tailings died . Field plant diversity was lower in amended areas than in CVA or UUC, with a higher percentage of the vegetative cover consisting of grasses . Small mammal analysis showed a low potential for elevated body Cd and Pb in the amended tailings . A re-entrainment study using fathead minnows (Pimephales promelas) showed no danger for resuspended amended tailings, as survival of fish was similar to the laboratory control . Data suggest that ecosystem function has been restored to the amended tailings, but that these systems are not yet in equilibrium. J Environ Qual, 2005 Jan-Feb, 34(1), 29 - 41 Beneficial reuse and sustainability: the fate of organic compounds in land-applied waste; Overcash M et al.; Land application systems, also referred to as beneficial reuse systems, are engineered systems that have defined and permitted application areas based on site and waste characteristics to determine the land area size requirement . These terrestrial systems have orders of magnitude greater microbial capability and residence time to achieve decomposition and assimilation compared with aquatic systems . In this paper we focus on current information and information needs related to terrestrial fate pathways in land treatment systems . Attention is given to conventional organic chemicals as well as new estrogenic and pharmaceutical chemicals of commerce . Specific terrestrial fate pathways addressed include: decomposition, bound residue formation, leaching, runoff, and crop uptake . Molecular decomposition and formation of bound residues provide the basis for the design and regulation of land treatment systems . These mechanisms allow for assimilation of wastes and nondegradation of the environment and accomplish the goal of sustainable land use . Bound residues that are biologically produced are relatively immobile, degrade at rates similar to natural soil materials, and should present a significantly reduced risk to the environment as opposed to parent contaminants . With regard to leaching and runoff pathways, no comprehensive summary or mathematical model of organic chemical migration from land treatment systems has been developed . For the crop uptake pathway, a critical need exists to develop information for nonagricultural chemicals and to address full-scale performance and monitoring at more land application sites . The limited technology choices for treatment of biosolids, liquids, and other wastes implies that acceptance of some risks and occurrence of some benefits will continue to characterize land application practices that contribute directly to the goal of beneficial reuse and sustainability. Ann Rheum Dis, 2005 Feb, 64(2), 279 - 285 Chronic non-bacterial osteomyelitis in children; Girschick HJ et al.; BACKGROUND: Chronic recurrent multifocal osteomyelitis (CRMO) in children is a chronic non-suppurative inflammation involving multiple sites . Some children affected by chronic non-bacterial osteomyelitis (CNO) do not have multiple lesions or a recurrent course . OBJECTIVE: To characterise the long term outcome of children with the full spectrum of CNO . METHODS: 30 children diagnosed with CNO were followed up for a mean of 5.6 years and their disease assessed using a clinical score, multiple imaging, and a diagnostic biopsy, including extensive microbial analysis . RESULTS: 9 patients had unifocal non-relapsing disease, 3 unifocal lesions with relapses, 9 multifocal lesions without relapses, and 9 multifocal lesions with relapses (CRMO) . Granulocytes were present significantly more often in CRMO than in unifocal and non-recurrent lesions . Pustulosis was more common in multifocal cases regardless of recurrence . Mean duration of treatment in 15 children with a single occurrence was 9.2 months . Naproxen treatment was generally effective . Naproxen treatment in 12 patients with relapses lasted 25 months . However, 7 of these were not effectively treated with naproxen alone . Five were treated with oral glucocorticoids for 27 days in addition to naproxen, which induced remission in four, lasting for at least 1.5 years . Longitudinal growth of affected bones was not altered, except for the development of hyperostosis . CONCLUSION: CNO is a spectrum of inflammatory conditions, with CRMO being the most severe . Most children with CNO have a favourable outcome of the disease . Oral glucocorticoids may be necessary in severe recurrent cases. J Biol Chem . 2005 Jan 12; {Epub ahead of print} A central role for the Hsp90-Cdc37 molecular chaperone module in IRAK-1-dependent signaling by toll-like receptors; Denardo D et al.; Toll-like receptors (TLRs) serve crucial roles in innate immunity by mediating the activation of macrophages by microbial pathogens . The protein kinase IRAK-1 is a key component of TLR signaling pathways via its interaction with TRAF6, which subsequently leads to the activation of MAP kinases and various transcription factors . IRAK-1 is degraded following TLR activation and this has been proposed to contribute to tolerance in macrophages by limiting further TLR-mediated signaling . Using a mass spectrometric-based approach we have identified a cohort of chaperones and co-chaperones including Hsp90 and Cdc37, which bind to IRAK-1 but not IRAK-4 in 293T cells . Pharmacologic inhibition of Hsp90 led to a rapid decline in the expression level of IRAK-1, whereas overexpression of Cdc37 enhanced the activation and oligomerization of IRAK-1 in 293T cells . Significantly, inhibition of Hsp90 in macrophages resulted in the destabilization and degradation of IRAK-1 but not IRAK-4 . Concomitant with the loss of IRAK-1 expression was a reduction in the activation of p38 MAP kinase and Erk1/2 following stimulation with the bacterially-derived TLR ligands lipopolysaccharide and CpG DNA . Moreover, TLR ligand-induced expression of proinflammatory cytokines was also reduced . Thus we conclude that the level of on-going support provided to IRAK-1 by the Hsp90-Cdc37 chaperone module directly influences the magnitude of TLR-mediated macrophage activation . In addition, because further TLR signaling depends on the synthesis of new IRAK-1, the Hsp90-Cdc37 chaperone module could also contribute to tolerance in macrophages by controlling the rate at which nascent IRAK-1 is folded into a functional conformation. Allergy, 2005 Feb, 60(2), 177 - 84 Exposure to pets, and the association with hay fever, asthma, and atopic sensitization in rural children; Waser M et al.; Background: An increasing number of studies report pet exposure to be associated with lower risk of asthma and allergies . This 'protective pet effect' has been suggested to result from a modified T-helper (Th)2-cell response, or because of increased microbial load in homes where pets are kept . We examined the associations between pet contact and the occurrence of asthma and allergies in children of the rural Allergy and Endotoxin (ALEX) population, taking farm animal contact, endotoxin and cat allergen levels in mattress dust into account . Methods: Information about contact with pets and farm animals, asthma and allergy were collected for 812 children by a standardized parents' questionnaire and an interview . Mattress dust endotoxin and cat allergen levels as well as specific IgE and IgG4 antibodies to Fel d1 were determined . Results: Current contact with dogs was inversely associated with diagnosed hay fever (OR 0.26, 95% CI 0.11-0.57), diagnosed asthma (OR 0.29, 95% CI 0.12-0.71), sensitization to cat allergen (OR 0.48, 95% CI 0.23-0.99) and to grass pollen (OR 0.55, 95% CI 0.33-0.94), but not with increased IgG4 levels . Early and current contact with cats were associated with reduced risk of wheezing (OR 0.48, 95% CI 0.23-1.00, and OR 0.49, 95% CI 0.26-0.92, respectively) and grass pollen sensitization . Adjustment for farm animal contact but not for endotoxin and cat allergen exposure attenuated these associations and the effect of pet was stronger among farmers' children . Conclusion: Although pet exposure was very frequent in this rural population, the inverse relation between current dog contact, asthma and allergy was mostly explained by simultaneously occurring exposure to stable animals or was restricted to farm children . In addition, a subtle form of pet avoidance may contribute to the protective effect of pet. Crit Rev Microbiol, 2004, 30(4), 263 - 73 Stress and how bacteria cope with death and survival; Aertsen A et al.; Bacterial populations that are exposed to rapidly changing and sometimes hostile environments constantly switch between growth, survival, and death . Understanding bacterial survival and death are therefore cornerstones in a full comprehension of microbial life . During the last few years, new insights have emerged regarding the mechanisms of bacterial inactivation under stressful conditions . Particularly under mildly lethal stress, the ultimate cause of inactivation often seems mediated by the cell itself and is subject to additional regulation that integrates information about the global state of the cell and its environmental and social surrounding . This article explores the thin line between bacterial growth and inactivation and focuses on some emerging bacterial survival strategies, both from an individual cell and from a population perspective. Can J Microbiol, 2004 Sep, 50(9), 745 - 750 Influence of carbon and nitrogen sources on growth, nitrogenase activity, and carbon metabolism of Gluconacetobacter diazotrophicus; Tejera NA et al.; The effects of different carbon and nitrogen sources on the growth, nitrogenase activity, and carbon metabolism of Gluconacetobacter diazotrophicus were investigated . The amino acids asparagine, aspartic acid, and glutamic acid affected microbial growth and nitrogenase activity . Several enzymatic activities involved in the tricarboxylic acid cycle were affected by the carbon source used . In addition, glucose and gluconate significantly increased the oxygen consumption (respiration rate) of whole cells of G . diazotrophicus grown under aerobic conditions . Enzymes responsible for direct oxidation of glucose and gluconate were especially active in cells grown with sucrose and gluconate . The presence of amino acids in the apoplastic and symplastic sap of sugarcane stems suggests that these compounds might be of importance in the regulation of growth and nitrogenase activity during the symbiotic association . The information obtained from the plant&ndash;bacterium association together with the results of other biochemical studies could contribute to the development of biotechnological applications of G . diazotrophicus. J Anim Sci, 2005 Feb, 83(2), 376 - 85 Effect of microbial phytase addition with or without the trace mineral premix in nursery, growing, and finishing pig diets; Shelton JL et al.; Two experiments were conducted to determine the interactive effects of phytase with and without a trace mineral premix (TMP) in diets for nursery, growing, and finishing pigs on growth performance, bone responses, and tissue mineral concentrations . Pigs (initial and final BW of 5.5 and 111.6 kg {Exp . 1} or 5.4 and 22.6 kg {Exp . 2}) were allotted to treatments on the basis of BW with eight (Exp . 1) or six (Exp . 2) replications of six or seven pigs per replicate pen . Pigs were started on the diets the day of weaning (average of 18 d) . In both experiments, the treatments were with or without 500 phytase units/kg of diet and with or without the TMP in a 2 x 2 factorial arrangement . The Ca and available P concentrations were decreased by 0.10% in diets with phytase . The nursery phase consisted of Phase I (7 d), Phase II (14 d), and Phase III (13 d) periods . In Exp . 1, 26 of 52 pigs fed the diet without the TMP and without phytase had severe skin lesions and decreased growth performance; therefore, pigs fed this diet were switched to the positive control diet . In Exp . 2, the treatment without the TMP and without phytase had 12 replications instead of six . At the end of Phase III, half these replications were switched to the positive control diet and half were switched to the diet without the TMP but with phytase . In Exp . 1 during Phases II and III and in the overall data, pigs fed the diet without the TMP had decreased ADG and ADFI, but the addition of phytase prevented these responses (phytase x TMP; P < 0.02) . Growth performance was not affected by diet during the growing-finishing period . Coccygeal bone Zn and Na concentrations were decreased (P < 0.09) in pigs fed the diet without the TMP, and adding phytase increased (P < 0.03) Zn and Fe concentrations . In Exp . 2 during Phases I and II, pigs fed the diet without the TMP had decreased ADG, but the addition of phytase prevented this response (phytase x TMP; P < 0.10) . Pigs fed the diet without the TMP had decreased (P < 0.10) ADG (Phase II and overall), ADFI (Phases II and III and in the overall data), and G:F (Phase III) . Coccygeal bone Zn and Cu concentrations were decreased (P < 0.09) in pigs fed the diet without the TMP, and adding phytase increased (P < 0.03) Zn concentration in the bones . These data indicate that removing the TMP in diets for nursery pigs decreases growth performance and bone mineral content, and that phytase addition to the diet without the TMP prevented the decreased growth performance. Water Res, 2005 Jan-Feb, 39(2-3), 494 - 500 Epub 2004 Dec 20. Sediment treatment with a nitrate-storing compound to reduce phosphorus release; Wauer G et al.; The application of Fe(III), in combination with sediment oxidation by NO(3)(-), is an accepted procedure to manage stratified eutrophic lakes by controlling the phosphorus release from sediments into overlying water . Depox(R), a newly developed compound, consisting of Fe(III) and NO(3)(-), has a storage effect for NO(3)(-) . NO(3)(-) is released slowly, hence the disadvantageous high solubility of NO(3)(-) in water can be retarded . The compound was added to water as a suspension which quickly flocculated and precipitated . Within 3 weeks, NO(3)(-) was desorbed from the Depox(R) compound in deionized water . After application in lakes, the NO(3)(-) availability on the sediment surface was prolonged for 2 months . After treatment, P release from the sediment and microbial metabolism were investigated under laboratory conditions as well as in the mesocosm . P release was almost stopped in both cases during the experiment . SO(4)(2-) consumption was significantly lower after Depox(R) addition, and CH(4) production was completely suppressed by Depox(R) treatment in the laboratory, whereas in the enclosures SO(4)(2-) and also CH(4) concentrations at the sediment water interface did not change significantly between treated enclosures and controls. Lett Appl Microbiol, 2005, 40(2), 128 - 32 Rapid monitoring of microbial contamination on herbal medicines by fluorescent staining method; Nakajima K et al.; Abstract k . nakajima, k . nonaka, k . yamamoto, n . yamaguchi, k . tani and m . nasu . 2004.Aims: To apply fluorescent staining method for fast assessment of microbial quality of herbal medicines . Methods and Results: The number of total bacteria and esterase-active bacteria on powdered traditional Chinese medicines were enumerated by fluorescent staining method using 6-carboxyfluorescein diacetate (6CFDA) and 4',6-diamidino-2-phenylindole (DAPI), and they were compared with colony-forming units (CFU) . The CFU was approx . 10(3) per gram in ginseng radix, and no bacterial colonies were detected from others . However, the total bacterial number (TDC) was more than 10(7) per gram, and number of bacteria possessing esterase activity ranged from 1 to 3% of TDC . Conclusions, Significance and Impact of the Study: Many bacteria in each Chinese medicine had enzyme activity and most of them could not be detected by conventional plate counting technique . Enumeration of bacterial cells on traditional Chinese medicines by fluorescent staining method requires less than 1 h . The double staining method with 6CFDA and DAPI could be applicable to rapid microbial monitoring of crude drugs. J Clin Periodontol, 2005 Jan, 32(1), 75 - 80 Anti-gingivitis efficacy of a stabilized 0.454% stannous fluoride/sodium hexametaphosphate dentifrice; Mankodi S et al.; Mankodi S, Bartizek RD, Winston JL, Biesbrock AR, McClanahan SF, He T: Anti-gingivitis efficacy of a stabilized 0.454% stannous fluoride/sodium hexametaphosphate dentifrice: a controlled six-month clinical trial . J Clin Peridontol 2004; doi: 10.1111/j.1600-051X.2004.00639.x . (c) Blackwell Munksgaard, 2004 . Abstract Objective: Stannous fluoride is a broad-spectrum anti-microbial agent that has been used in dentistry as a chemical adjunct to prevent dental caries and gingivitis . The objective of this study was to assess the anti-gingivitis efficacy of a stabilized 0.454% stannous fluoride/sodium hexametaphosphate dentifrice relative to a negative control . Methods: This was a randomized, 6-month, double-blind, parallel-group gingivitis study conducted according to the guidelines for evaluating chemotherapeutic products for the control of gingivitis outlined by the American Dental Association . A stabilized 0.454% stannous fluoride/sodium hexametaphosphate dentifrice was tested against a commercially available negative control dentifrice . Following baseline measurements, subjects received a dental prophylaxis . Subjects were then instructed to brush twice daily for 60 s using their assigned product . Oral soft-hard-tissue examinations and clinical examinations using the Modified Gingival Index, Gingival Bleeding Index, and the Turesky modification of the Quigley-Hein Plaque Index were performed at baseline, 3 and 6 months post-treatment . Results: A total of 143 subjects were enrolled and 130 of them completed the 6-month study . After 6 months of product usage, the experimental group had 21.7% less gingivitis (p<0.001), 57.1% less bleeding (p<0.001), and 6.9% less plaque (p=0.01) on average compared with the negative control group . No adverse oral soft-hard-tissue effects or extrinsic tooth staining was observed in the study . Conclusion: The results demonstrate that use of the stabilized 0.454% stannous fluoride/sodium hexametaphosphate dentifrice over a 6-month period provided statistically significant reductions in gingivitis, gingival bleeding, and plaque when compared with a negative control dentifrice. Langmuir, 2005 Jan 18, 21(2), 530 - 536 Liposome Fluidization and Melting Point Depression by Pressurized CO(2) Determined by Fluorescence Anisotropy; Bothun GD et al.; The influence of CO(2) on the bilayer fluidity of liposomes, which are representative of model cellular membranes, was examined for the first time at the elevated pressures (up to 13.9 MPa) associated with CO(2)-based processing of liposomes and microbial sterilization . Fluidization and melting point depression of aqueous dipalmitoylphosphatidylcholine (DPPC) liposomes by pressurized CO(2) (present as an excess phase) were studied by steady-state fluorescence anisotropy using the membrane probe 1,6-diphenyl-1,3,5-hexatriene (DPH) . Isothermal experiments revealed reversible, pressure-dependent fluidization of DPPC bilayers at temperatures corresponding to near-gel (295 K) and fluid (333 K) phases at atmospheric pressure, where the gel-to-fluid phase transition (T(m)) occurs at approximately 315 K . Isobaric measurements (P(CO)()2 =1.8, 7.0, and 13.9 MPa) of DPH anisotropy demonstrate substantial melting point depression (DeltaT(m) = -4.8 to -18.5 K) and a large broadening of the gel-fluid phase transition region, which were interpreted using conventional theories of melting point depression . Liposome fluidity is influenced by CO(2) accumulation in the hydrocarbon core and polar headgroup region, as well as the formation of carbonic acid and/or the presence of buffering species under elevated CO(2) pressure. Biochemistry, 2005 Jan 18, 44(2), 726 - 733 Identification of Functional Amino Acids in the Nramp Family by a Combination of Evolutionary Analysis and Biophysical Studies of Metal and Proton Cotransport in Vivo; Chaloupka R et al.; The natural resistance-associated macrophage protein (Nramp) family is functionally conserved in bacteria and eukarya; Nramp homologues function as proton-dependent membrane transporters of divalent metals . Sequence analyses indicate that five phylogenetic groups comprise the Nramp family, three bacterial and two eukaryotic, which are distinct from a more distantly related group of microbial sequences (Nramp outgroup) . The Nramp family and outgroup share many conserved residues, suggesting they derived from a common ancestor and raising the possibility that the residues invariant in the Nramp family that correspond to residues which are different but also conserved in the outgroup represent candidate sites of functional divergence of the Nramp family . Four Nramp family-specific residues were identified within transmembrane domains 1, 6, and 11, and replaced by the corresponding invariant outgroup residues in the Escherichia coli Nramp ortholog (the proton-dependent manganese transporter, MntH of group A, EcoliA) . The resulting mutants (Asp(34)Gly, Asn(37)Thr, His(211)Tyr, and Asn(401)Gly) were tested for both divalent metal uptake and proton transport; quasi-simultaneous analyses of uptake of metals and protons revealed for the first time protons and metals cotransport by a bacterial Nramp homologue . Additional mutations were studied for comparison (Asp(34)Asn, Asn(37)Asp and Asn(37)Val, Asn(401)Thr, His(211)Ala, His(216)Ala, and His(216)Arg) . EcoliA activity was impaired after each of the Nramp/outgroup substitutions, as well as after more conservative replacements, showing that the tested sites are all important for metal uptake and metal-dependent H(+) transport . It is proposed that co-occurrence of these four Nramp-specific transmembrane residues may have contributed to the emergence of this family of metal and proton cotransporters. Compend Contin Educ Dent, 2004 Jan, 25(1 Suppl), 30, 32 - 34, 36-37 Environmental surface infection control, 2003; Molinari JA et al.; Infection control guidelines address the routine use of disposable covers and cleaning and disinfecting treatment-area surfaces contaminated during patient care . While the rationale and approaches for accomplishing environmental surface disinfection have been considered in many previous infection control recommendations, exaggerated claims have led to some confusion regarding the wide variety of available surface covers and disinfectant products . The Centers for Disease Control and Prevention Guideline for Infection Control in Dental Health-Care Settings, 2003, considers the current state of scientific and clinical knowledge in environmental infection control, reinforce basic principles, and provide updated recommendations aimed at minimizing the potential for microbial cross-contamination and cross-infection from inanimate surfaces. Compend Contin Educ Dent, 2004 Jan, 25(1 Suppl), 17 - 22 An immunization program for dental practices; Palenik CJ et al.; Immunization helps prevent acquisition of diseases, many of which were once common in the United States . Immunity is generally achieved by administering live or dead pathogens or their components . Vaccines used for active immunization consist of live (attenuated) organisms, killed whole or split organisms, microbial components, or inactivated toxins (toxoids) . Some cases require the administration of antibody-containing gamma globulin injections . According to the new Centers for Disease Control and Prevention Guideline on infection control, dental practices must develop a written, comprehensive policy on immunizing workers and refer personnel to qualified professionals for evaluation and, when necessary, for all appropriate immunizations. Neoplasma, 2004, 51(6), 407 - 14 Taraxasterol and beta-sitosterol: new naturally compounds with chemoprotective/chemopreventive effects; Ovesna Z et al.; Substantial attention has been given to primary cancer prevention in daily life . Dietary factors are through to contribute to as much as one-third of the factors influencing the development of cancer . Ones of the components of a plant-based diet are beta- sitosterol and taraxasterol, compounds attracting our specific attention . This review summarizes the biological activities of presented phytosterols (anti-inflammatory, cholesterol-lowering, anti-microbial, anti-bacterial, anti-fungal effects) . Our interest has been focussed especially on their anti-tumor and chemopreventive activity . They have been shown experimentally to inhibit colon and breast cancer development . They act at various stages of tumor development, including inhibition of tumorigenesis, inhibition of tumor promotion, and induction of cell differentiation . They effectively inhibit invasion of tumor cells and metastasis . With regard to toxicity, no obvious side effects of phytosterols have been observed in studies to date, with the exception of individuals with phytosterolemia . The exact mechanism by which dietary phytosterols act is not fully understood . However, some mechanisms have been offered . Therefore, they have a bright future in clinical application . Further investigation to explore their potential in tumor treatment may prove to be worthwhile. Curr Opin Allergy Clin Immunol, 2004 Dec, 4(6), 555 - 61 Toll-like receptor ligands: hygiene, atopy and therapeutic implications; Horner AA et al.; PURPOSE OF REVIEW: Allergic and certain other inflammatory diseases have become more common in industrialized countries over the past few decades . One potential explanation for such trends is that with a decreased incidence of microbial exposures, as a result of modern public health practices, an important source of immune stimulation has been lost, with a consequent increase in inflammatory responses and their associated diseases . This review will focus on our current understanding of how microbial exposures impact on host immunity and the pathogenesis of allergic diseases . RECENT FINDINGS: In the past decade, it has become clear that a number of molecular interactions between immunocytes and microbial compounds are mediated by Toll-like receptors on host cells . Moreover, recent investigations have suggested that ligands for different Toll-like receptors have the potential both to inhibit and promote the development of allergic hypersensitivities and diseases . SUMMARY: On the basis of studies discussed herein, we speculate that physiological exposures to Toll-like receptor ligands have important yet complex effects on immune homeostasis and host susceptibility towards atopic diseases . Moreover, we anticipate that a fuller understanding of how physiological Toll-like receptor ligand exposures impact on immune development will lead to novel therapeutic interventions for the prevention and treatment of atopic diseases. Appl Environ Microbiol, 2005 Jan, 71(1), 519 - 29 Characterization of Ichthyocidal Activity of Pfiesteria piscicida: Dependence on the Dinospore Cell Density; Drgon T et al.; The ichthyocidal activity of Pfiesteria piscicida dinospores was examined in an aquarium bioassay format by exposing fish to either Pfiesteria-containing environmental sediments or clonal P . piscicida . The presence of Pfiesteria spp . and the complexity of the microbial assemblage in the bioassay were assessed by molecular approaches . Cell-free water from bioassays that yielded significant fish mortality failed to show ichthyocidal activity . Histopathological examination of moribund and dead fish failed to reveal the skin lesions reported elsewhere . Fish larvae within "cages" of variable mesh sizes were killed in those where the pore size exceeded that of Pfiesteria dinospores . In vitro exposure of fish larvae to clonal P . piscicida indicated that fish mortality was directly proportional to the dinospore cell density . Dinospores clustered around the mouth, eyes, and operculi, suggesting that fish health may be affected by their direct interaction with skin, gill epithelia, or mucous surfaces . Molecular fingerprinting revealed the presence of a very diverse microbial community of bacteria, protists, and fungi within bioassay aquaria containing environmental sediments . Some components of the microbial community were identified as potential fish pathogens, preventing the rigorous identification of Pfiesteria spp . as the only cause of fish death . In summary, our results strongly suggest (i) that this aquarium bioassay format, which has been extensively reported in the literature, is unsuitable to accurately assess the ichthyocidal activity of Pfiesteria spp . and (ii) that the ichthyocidal activity of Pfiesteria spp . is mostly due to direct interactions of the zoospores with fish skin and gill epithelia rather than to soluble factors. Appl Environ Microbiol, 2005 Jan, 71(1), 467 - 79 Diversity and distribution of methanotrophic archaea at cold seeps; Knittel K et al.; In this study we investigated by using 16S rRNA-based methods the distribution and biomass of archaea in samples from (i) sediments above outcropping methane hydrate at Hydrate Ridge (Cascadia margin off Oregon) and (ii) massive microbial mats enclosing carbonate reefs (Crimea area, Black Sea) . The archaeal diversity was low in both locations; there were only four (Hydrate Ridge) and five (Black Sea) different phylogenetic clusters of sequences, most of which belonged to the methanotrophic archaea (ANME) . ANME group 2 (ANME-2) sequences were the most abundant and diverse sequences at Hydrate Ridge, whereas ANME-1 sequences dominated the Black Sea mats . Other seep-specific sequences belonged to the newly defined group ANME-3 (related to Methanococcoides spp.) and to the Crenarchaeota of marine benthic group B . Quantitative analysis of the samples by fluorescence in situ hybridization (FISH) showed that ANME-1 and ANME-2 co-occurred at the cold seep sites investigated . At Hydrate Ridge the surface sediments were dominated by aggregates consisting of ANME-2 and members of the Desulfosarcina-Desulfococcus branch (DSS) (ANME-2/DSS aggregates), which accounted for >90% of the total cell biomass . The numbers of ANME-1 cells increased strongly with depth; these cells accounted 1% of all single cells at the surface and more than 30% of all single cells (5% of the total cells) in 7- to 10-cm sediment horizons that were directly above layers of gas hydrate . In the Black Sea microbial mats ANME-1 accounted for about 50% of all cells . ANME-2/DSS aggregates occurred in microenvironments within the mat but accounted for only 1% of the total cells . FISH probes for the ANME-2a and ANME-2c subclusters were designed based on a comparative 16S rRNA analysis . In Hydrate Ridge sediments ANME-2a/DSS and ANME-2c/DSS aggregates differed significantly in morphology and abundance . The relative abundance values for these subgroups were remarkably different at Beggiatoa sites (80% ANME-2a, 20% ANME-2c) and Calyptogena sites (20% ANME-2a, 80% ANME-2c), indicating that there was preferential selection of the groups in the two habitats . These variations in the distribution, diversity, and morphology of methanotrophic consortia are discussed with respect to the presence of microbial ecotypes, niche formation, and biogeography. Appl Environ Microbiol, 2005 Jan, 71(1), 344 - 53 Roseobacter-like bacteria in red and mediterranean sea aerobic anoxygenic photosynthetic populations; Oz A et al.; Bacteriochlorophyll a-containing aerobic anoxygenic phototrophs (AAnP) have been proposed to account for up to 11% of the total surface water microbial community and to potentially have great ecological importance in the world's oceans . Recently, environmental and genomic data based on analysis of the pufM gene identified the existence of alpha-proteobacteria as well as possible gamma-like proteobacteria among AAnP in the Pacific Ocean . Here we report on analyses of environmental samples from the Red and Mediterranean Seas by using pufM as well as the bchX and bchL genes as molecular markers . The majority of photosynthesis genes retrieved from these seas were related to Roseobacter-like AAnP sequences . Furthermore, the sequence of a novel photosynthetic operon organization from an uncultured Roseobacter-like bacterial artificial chromosome retrieved from the Red Sea is described . The data show the presence of Roseobacter-like bacteria in Red and Mediterranean Sea AAnP populations in the seasons analyzed. Appl Environ Microbiol, 2005 Jan, 71(1), 276 - 81 Microbial Desulfurization of Gasoline in a Mycobacterium goodii X7B Immobilized-Cell System; Li F et al.; Mycobacterium goodii X7B, which had been primarily isolated as a bacterial strain capable of desulfurizing dibenzothiophene to produce 2-hydroxybiphenyl via the 4S pathway, was also found to desulfurize benzothiophene . The desulfurization product was identified as o-hydroxystyrene by gas chromatography (GC)-mass spectrometry analysis . This strain appeared to have the ability to remove organic sulfur from a broad range of sulfur species in gasoline . When Dushanzi straight-run gasoline (DSRG227) containing various organic sulfur compounds was treated with immobilized cells of strain X7B for 24 h, the total sulfur content significantly decreased, from 227 to 71 ppm at 40 degrees C . GC flame ionization detection and GC atomic emission detection analysis were used to qualitatively evaluate the effects of M . goodii X7B treatment on the contents of gasoline . In addition, when immobilized cells were incubated at 40 degrees C with DSRG275, the sulfur content decreased from 275 to 54 ppm in two consecutive reactions . With this excellent efficiency, strain X7B is considered a good potential candidate for industrial applications for the biodesulfurization of gasoline. Appl Environ Microbiol, 2005 Jan, 71(1), 227 - 39 Geographic and environmental sources of variation in lake bacterial community composition; Yannarell AC et al.; This study used a genetic fingerprinting technique (automated ribosomal intergenic spacer analysis {ARISA}) to characterize microbial communities from a culture-independent perspective and to identify those environmental factors that influence the diversity of bacterial assemblages in Wisconsin lakes . The relationships between bacterial community composition and 11 environmental variables for a suite of 30 lakes from northern and southern Wisconsin were explored by canonical correspondence analysis (CCA) . In addition, the study assessed the influences of ARISA fragment detection threshold (sensitivity) and the quantitative, semiquantitative, and binary (presence-absence) use of ARISA data . It was determined that the sensitivity of ARISA was influential only when presence-absence-transformed data were used . The outcomes of analyses depended somewhat on the data transformation applied to ARISA data, but there were some features common to all of the CCA models . These commonalities indicated that differences in bacterial communities were best explained by regional (i.e., northern versus southern Wisconsin lakes) and landscape level (i.e., seepage lakes versus drainage lakes) factors . ARISA profiles from May samples were consistently different from those collected in other months . In addition, communities varied along gradients of pH and water clarity (Secchi depth) both within and among regions . The results demonstrate that environmental, temporal, regional, and landscape level features interact to determine the makeup of bacterial assemblages in northern temperate lakes. Infect Genet Evol, 2005 Mar, 5(2), 103 - 8 Exploring microbial microevolution with microarrays; Ochman H et al.; Gene arrays are typically employed to monitor gene expression and regulation, but they are finding additional applications in studying patterns of evolution in bacterial genomes . In particular, this approach has been applied to answer questions about the heterogeneity in full gene repertoires among bacterial strains and species without relying on more costly and time-consuming methodologies . In this review, we evaluate some of the evolutionary patterns and processes affecting bacterial genomes as detected with microarrays, and also delineate the limitations and conclusions stemming from such studies. Clin Immunol, 2005 Feb, 114(2), 110 - 8 Pristane-induced autoimmunity in germ-free mice; Mizutani A et al.; Hypergammaglobulinemia and autoantibodies are reduced in pristane-treated specific pathogen-free mice vs . conventionally housed controls, consistent with the role of microbial stimulation in this model . To determine whether microbial stimulation is required, BALB/c mice housed under germ-free conditions were treated i.p . with sterile PBS or pristane and examined 6 months later . As in conventional mice, pristane-treated germ-free mice developed peritoneal granulomas and hypergammaglobulinemia with increased IgG2a/IgG1 ratios . LPS stimulation induced more IL-6, IL-12, and TNF-alpha, and anti-CD3 induced more IFN-gamma and IL-4 by peritoneal cells from pristane-treated mice vs . control . Anti-nRNP/Sm and -Su autoantibodies were found in 40% and 43%, respectively, of pristane-treated germ-free mice by immunoprecipitation . Thus, bacterial stimulation was not required for lupus autoantibodies, peritoneal granuloma formation, hypergammaglobulinemia, or cytokine overproduction . Although microbial stimulation acts synergistically with pristane, these results clearly indicate that pristane does not act merely by increasing exposure to microbial products such as LPS. Forensic Sci Int, 2004 Dec 2, 146 Suppl, S185 - 8 Genetics and attribution issues that confront the microbial forensics field; Budowle B; The commission of an act of bioterrorism or biocrime is a real concern for law enforcement and society . Efforts are underway to develop a strong microbial forensic program to assist in identifying perpetrators of acts of bioterrorism and biocrimes, as well as serve as a deterrent for those who might commit such illicit acts . Genetic analyses of microbial organisms will likely be a powerful tool for attribution of criminal acts . There are some similarities to forensic human DNA analysis practices, such as: molecular biology technology, use of population databases, qualitative conclusions of test results, and the application of QA/QC practices . Differences include: database size and composition, statistical interpretation methods, and confidence/uncertainty in the outcome of an interpretation. J Biotechnol, 2005 Feb 23, 115(4), 397 - 404 Epub 2004 Nov 11. Morphological and physiological changes in Saccharomyces cerevisiae by oxidative stress from hyperbaric air; Belo I et al.; Increase in air or oxygen pressure in microbial cell cultures can cause oxidative stress and consequently affect cell physiology and morphology . The behaviour of Saccharomyces cerevisiae grown under hyperbaric atmospheres of air and pure oxygen was studied . A limit of 1.0MPa for the air pressure increase (i.e . 0.21MPa of oxygen partial pressure) in a fed-batch culture of S . cerevisiae was established . Values of 1.5MPa air pressure and 0.32MPa pure oxygen pressure strongly inhibited the metabolic activity and the viability of the cells . Also, morphological changes were observed, especially cell-size distribution and the genealogical age profile . Pressure caused cell compression and an increase in number of aged cells . These effects were attributed to oxygen toxicity since similar results were obtained using air or oxygen, if oxygen partial pressure was equal to or higher than 0.32MPa . The activity of the antioxidant enzymes, catalase and superoxide dismutase (SOD) (cytosolic and mitochondrial isoformes) indicated that the enzymes have different roles in oxidative stress cell protection, depending on other factors that affect the cell physiological state. Mini Rev Med Chem, 2005 Jan, 5(1), 57 - 71 Pharmacological properties of furoxans and benzofuroxans: recent developments; Cerecetto H et al.; The chemistry of furoxans (1, 2, 5-oxadiazole-2-oxides) and benzofuroxans (benzo{1, 2-c}1, 2, 5-oxadiazole-1-oxides) is very well known . These systems are widely used in organic chemistry as intermediate compounds for the synthesis of numerous heterocycles . In the other hand, furoxan and benzofuroxan derivatives were extensively studied as bioactive compounds . They possess remarkable biological activities, such as anti-microbial and anti-parasitic properties, mutagenic, immunosuppressive and anticancer effects, anti-aggregating and vasorelaxant activity, among others . In some cases, molecular mode of action was proposed . Recently, the research and development in the medicinal chemistry of these systems have produced hybrid compounds in which furoxan or benzofuroxan moieties together with a classical drug moieties are present in a single molecule . So, new anti-ulcer drugs, calcium channel modulators and vasodilator derivatives were described and they are currently in study . In this presentation recent developments in the medicinal chemistry of furoxans and benzofuroxans will be reviewed. Biomacromolecules, 2005 Jan-Feb, 6(1), 35 - 8 Transglutaminase-mediated protein immobilization to casein nanolayers created on a plastic surface; Kamiya N et al.; An enzymatic method for covalent and site-specific immobilization of recombinant proteins on a plastic surface was explored . Using Escherichia coli alkaline phosphatase (AP) with a specific peptide tag (MKHKGS) genetically incorporated at the N-terminus as a model (NK-AP), microbial transglutaminase (MTG)-mediated protein immobilization was demonstrated . To generate a reactive surface for MTG, a 96-well polystyrene microtiter plate was physically coated with casein, a good MTG substrate . Successful immobilization of recombinant AP to the nanolayer of casein on the surface of the microtiter plate was verified by the detection of enzymatic activity . Since little activity was observed when wild-type AP was used, immobilization of NK-AP was likely directed by the specific peptide tag . When polymeric casein prepared by MTG was used as a matrix on the plate, the loading capacity of AP was increased about 2-fold compared to when casein was used as the matrix . Transglutaminase-mediated site-specific posttranslational modification of proteins offers one way of generating a variety of protein-based solid formulations for biotechnological applications. J Allergy Clin Immunol, 2005 Jan, 115(1), 25 - 30 Prototypic disorders of gastrointestinal mucosal immune function: Celiac disease and Crohn's disease; James SP; Celiac disease is a disorder of the small intestine characterized by chronic inflammation of the mucosa and protean clinical manifestations caused by loss of tolerance to dietary antigens . Two strongly associated cofactors have been identified: the presence of HLA-DQ2 or HLA-DQ8 in the host and specific antigenic peptides in the diet that are present in wheat, rye, and barley . Most patients have complete remission after dietary elimination of these foods . Crohn's disease is characterized by chronic, relapsing, recurrent, focal, transmural inflammation of the gastrointestinal tract that can lead to multiple serious problems requiring chronic medical and surgical therapy . Crohn's disease is associated with multiple genetic mutations, at least one of which has been clearly implicated in innate immunity . Multiple lines of evidence suggest that the disease involves abnormal immune responses to gut microbial flora. Nutr Clin Care, 2004 Oct-Dec, 7(4), 134 - 40 Foodborne illness primer for physicians and other health care professionals; Foodborne Illness Primer Work Group; Foodborne illness is a serious public health problem . According to the Centers for Disease Control and Prevention (CDC), it is estimated that every year, 76 million Americans get sick, leading to over 300,000 hospitalizations and 5000 deaths . Recent outbreaks underscore Americans' vulnerability to foodborne pathogens . Primarily the very young, the elderly, and the immunocompromised are affected . Recent changes in human demographics and food preferences, changes in food production and distribution systems, and microbial adaptation have led to the emergence of novel as well as traditional foodborne diseases . With increasing travel and trade opportunities, it is not surprising that now there is a greater risk of contracting and spreading a foodborne illness locally, regionally, and even globally . To better recognize, diagnose, treat, and prevent foodborne illnesses, the Foodborne Illness Primer Working Group collaborated to publish the new, updated edition of Diagnosis and Management of Foodborne Illnesses: A Primer for Physicians and Other Health Care Professionals . Physicians and other health care professionals have a critical role in the prevention and control of food-related disease outbreaks . This primer is intended to provide practical and concise information on the diagnosis, treatment, reporting, and prevention of foodborne illnesses. Macromol Biosci, 2005 Jan 14, 5(1), 30 - 8 Comb-Like Ionic Complexes of Cationic Surfactants with Bacterial Poly(gamma-glutamic acid) of Racemic Composition; Garcia-Alvarez M et al.; Comb-like ionic complexes, nATMA . PG(DL)GA, were prepared from microbial poly(gamma-glutamic acid), with a nearly racemic configuration, and alkyltrimethylammoniun bromides, with linear alkyl chains containing an even number of carbon atoms, n, ranging from 12 to 22 . The complexes had a nearly stoichiometric composition, displayed thermal stability up to temperatures above 200 degrees C and were insoluble in water but soluble in organic solvents . In the solid state, they were arranged in a regularly layered structure with the alkyl side chain crystallized for n >/= 18 . Heating above melting entailed a contraction in the interlayer distance which varied from 1 to 10% depending on the value of n . Comparison with data reported on similar complexes obtained from nearly enantiomerically pure poly(gamma-glutamic acid) revealed an overall behavior very similar for the two series but with specific significant differences concerning side chain crystallinity and dimensional response to temperature. J Immunol, 2005 Jan 15, 174(2), 727 - 34 The reciprocal interaction of NK cells with plasmacytoid or myeloid dendritic cells profoundly affects innate resistance functions; Gerosa F et al.; A reciprocal activating interaction between NK cells and dendritic cells (DC) has been suggested to play a role in the functional regulation of these cells in immunity, but it has been studied only using in vitro generated bone marrow- or monocyte-derived DC . We report that human peripheral blood plasmacytoid DC (pDC) and myeloid DC are necessary to induce NK cell function depending on the type of microbial stimulus . pDC and myeloid DC are required for strongly increased NK cytolytic activity and CD69 expression, in response to inactivated influenza virus or CpG-containing oligonucleotides and poly(I:C), respectively . Secreted type I IFN is required and sufficient for the augmentation of NK cell cytolytic activity in the coculture with pDC or myeloid DC, whereas CD69 expression is dependent on both type I IFN and TNF . In addition, in response to poly(I:C), myeloid DC induce NK cells to produce IFN-gamma through a mechanism dependent on both IL-12 secretion and cell contact between NK cells and myeloid DC, but independent of type I IFN . IL-2-activated NK cells have little to no cytolytic activity for immature myeloid DC and pDC, but are able to induce maturation of these cells . Moreover, IL-2-activated NK cells induce, in the presence of a suboptimal concentration of CpG-containing oligonucleotides, a strong IFN-alpha and TNF production . These data suggest that the reciprocal functional interaction between NK cells and either pDC or myeloid DC may play an important physiological role in the regulation of both innate resistance and adaptive immunity to infections. J Immunol, 2005 Jan 15, 174(2), 589 - 94 Leukotrienes: underappreciated mediators of innate immune responses; Peters-Golden M et al.; Leukotrienes are bronchoconstrictor and vasoactive lipid mediators that are targets in the treatment of asthma . Although they are increasingly recognized to exert broad proinflammatory effects, their role in innate immune responses is less well appreciated . These molecules are indeed synthesized by resident and recruited leukocytes during infection . Acting via cell surface G protein-coupled receptors and subsequent intracellular signaling events, they enhance leukocyte accumulation, phagocyte capacity for microbial ingestion and killing, and generation of other proinflammatory mediators . Interestingly, a variety of acquired states of immunodeficiency, such as HIV infection and malnutrition, are characterized by a relative deficiency of leukotriene synthesis . The data reviewed herein point to leukotrienes as underappreciated yet highly relevant mediators of innate immunity. J Biol Chem . 2005 Jan 5; {Epub ahead of print} The sulfogalactose moiety of sulfoglycosphingolipids serves as a mimic of tyrosine phosphate in many recognition processes: prediction and demonstration of SH2 domain/sulfogalactose binding; Lingwood CA et al.; Multiple ligand co-recognition of 3'sulfogalactosylceramide(SGC) and sulfotyrosine initiated comparison of SGC and sulfotyrosine, and subsequently, phosphotyrosine(pY) binding . 3'Sulfogalactosylceramide(SGC) is a receptor for ligands involved in cell adhesion/microbial pathology . pY forms a Src homology domain(SH2) recognition motif in intracellular signaling . Using hsp70, anti-SGC and anti-pY antibodies , ligand binding is retained following phosphate/sulfate and tyrosine/galactose substitution in SGC and sulfate/phosphate exchange in pY . Remarkable lipid-dependent binding to phosphatidylethanolamine-conjugated sulfotyrosine, suggests 'microenvironmental' modulation of sulfotyrosine-containing receptors, similar to glycosphingolipids . Modeling, based on an aryl substrate-bound cocrystal of arylsulfatase A, a sulfogalactose and phosphotyrosine esterase, provides a solvation basis for co-recognition . c-SrcSH2 domain:SGC/phosphogalactosyl ceramide binding confirms our hypothesis, heralding a carbohydrate-based approach to regulation of phosphotyrosine-mediated recognition. Zhonghua Yan Ke Za Zhi, 2004 Nov, 40(11), 750 - 4 {Clinical study of the treatment of acanthamoeba keratitis by penetrating keratoplasty.}; Shi WY et al.; OBJECTIVE: To evaluate the influence of accurate diagnosis and peri-operative management on the recurrence of acanthamoeba keratitis after penetrating keratoplasty . METHODS: A retrospective study of 18 cases of medically unresponsive keratitis in Shandong Eye Institute and Hospital that were confirmed as acanthamoeba keratitis based on laboratory examination or histopathological studies . The diagnosis of acanthamoeba keratitis was made prior to the surgery in 12 cases and delayed until histopathological results in six cases . All cases ran a course of one to four months prior to penetrating keratoplasty with lesions measuring larger than 7 mm at the time of surgery . Corresponding anti-microbial agents were given pre-operatively based on the suspected etiological agents . Corticosteroids were given to two eyes diagnosed as acanthamoeba keratitis, three eyes misdiagnosed as herpes simplex keratits and two eyes misdiagnosed as bacterial keratitis . RESULTS: In 13 cases without recurrence (72%), 11 cases were correctly diagnosed as acanthamoeba keratitis prior to the surgery . In five cases with relapse (28%), four cases were misdiagnosised prior to the surgery . Seven cases received corticosteroid therapy more than one week in early post-operative period and four cases developed recurrence . In the four cases with relapse, two were misdiagnosed as herpes keratitis and two were misdiagnosed as bacterial keratitis . The odds ratis of the effect of two factors (misdiagnosis and using of corticosteroid in early post-operative period) on the rate of recurrence were 22 (P = 0.021) and 13 (P = 0.047), respectively . CONCLUSIONS: Penetrating keratoplasty is an effective treatment for medically unresponsive acanthamoeba keratitis . Accurate pre-operative diagnosis, appropriate peri-operative amoebicidal therapy and avoidance from the use of corticosteroid in the early post-operative period can decrease the incidence of recurrence of acanthamoeba keratitis after penetrating keratoplasty. J Food Prot, 2004 Dec, 67(12), 2809 - 11 A survey of microbial contamination of food contact surfaces at broiler slaughter plants in Taiwan; Ho CP et al.; Microbial contamination levels at broiler slaughter plants were investigated at three major slaughter plants in Taiwan during the summer and winter . The microbial contamination levels in chicken carcasses and on food contact surfaces were examined using the swab method . The results indicated that the bacterial counts were affected by the slaughter processing plant, processes, and season (P < 0.05) . The bacterial counts on food contact surfaces of the equipment before operation were not significantly lower than those after processing . Regardless of the bacterial type, bacterial counts of chicken carcasses generally decreased from the scalding step to the washing step before evisceration and then increased . The cleaning procedures for food contact surfaces should be evaluated, and special attention should be given to utensils used during processing, such as gloves, baskets, and hand tools. J Food Prot, 2004 Dec, 67(12), 2767 - 71 Menadione-catalyzed luminol chemiluminescence assay for the rapid detection of viable bacteria in foods under aerobic conditions; Kawasaki S et al.; A menadione-catalyzed luminol chemiluminescence assay was developed for the rapid detection and estimation of viable bacteria in foods . The principle of this assay is based on the extracellular menadione-catalyzed active oxygen spieces (O2- and H2O2) generated by the activity of NAD(P)H:menadione oxidoreductase in viable cells . This luminol chemiluminescence assay requires 10 min for the incubation of cells with menadione and then 2 s for the measurement of chemiluminescence intensity after an injection of luminol solution without the treatment of cell lysis . This method was evaluated using liquid food samples of milk, vegetable juice, green tea, and coffee spiked with Escherichia coli ATCC 25922 . The study result revealed that E . coli contamination at 1 to 10 CFU/ml in these foods could be detected after incubation at 37 degrees C for 7 h in an enrichment medium; however, the green tea and coffee samples requires filtration . This method could be a useful tool for the rapid evaluation of microbial food contamination. Int Dent J, 2004 Dec, 54(6), 438 - 44 An investigation of the microbial contamination of dental unit air and water lines; Walker RJ et al.; AIM: The objective of this study was to measure the microbial contamination released from dental unit air lines (DUAL) and dental unit water lines (DUWL) . MATERIALS AND METHODS: Emissions from DUAL and DUWL from five dental units supplied by a central water source (tap) and a centralised air supply were sampled three times over a five- week period . Air was forced through sterile water and then plated onto selective agar using apparatus designed to spread the sample solution evenly, and then incubated at room temperature for seven days . Colonies were then counted and the concentration of bacteria present was determined and expressed as colony forming units per millilitre (cfu/ml) per minute . The same procedure was used to evaluate five other dental units, which had attached independent water reservoir (bottle) systems (IWR) . Only deionised water was added to the bottles and unit waterlines were cleaned weekly with alkaline peroxide based solution . Values were expressed as cfu/ml . RESULTS: Air and water specimens obtained from dental units supplied with tap water had microbial counts significantly (t-test, p < 0.05) greater than IWR dental units . CONCLUSIONS: Results indicate that IWR can reduce the numbers of micro-organisms released from DUWL . However, the effectiveness of such systems should be routinely monitored . Emissions from DUAL seems to reflect the levels of microbial contamination present in DUWL. J Periodontol, 2004 Nov, 75(11), 1486 - 92 Commensal bacteria influence innate status within gingival tissues: a pilot study; Dixon DR et al.; BACKGROUND: The objective of this study was to determine the contribution of commensal bacteria to the innate defense status of gingival tissue by examining the expression of innate host defense mediators in germ-free and conventionally reared groups in both BALBc/ByJ and SCID C.B17 mice . METHODS: Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was utilized to determine the constitutive levels within each gingival tissue set (N = 5) for: E-selectin, P-selectin, interleukin-(IL)-8 homologue, tumor necrosis factor-alpha, IL-1beta, intercellular adhesion molecule-(ICAM)-1, ICAM-2, and vascular adhesion molecule-(VCAM)-1 . In addition, IL-1beta protein content was determined by enzyme-linked immunosorbent assay (ELISA) . RESULTS: Gingival samples revealed that only IL-1beta mRNA expression among all mediators examined was significantly reduced in conventionally reared mice (P<0.01) compared to germ-free mice . In contrast, IL-1beta protein levels were significantly (P <0.001) higher in conventionally reared mice compared to germ-free animals . Conventionally reared and germ-free SCID C.B17 mice revealed a similar pattern in regard to reduced IL-1beta mRNA and significantly increased IL-1beta protein (P<0.0001) . CONCLUSION: Commensal microbial colonization influences innate host defense mediator expression of IL-1beta at both the mRNA and protein levels in healthy periodontal tissue in mice. Bosn J Basic Med Sci, 2004 Oct, 4(4), 51 - 8 Formulation ingredients for toothpastes and mouthwashes; Vranic E et al.; In order to achieve the multi-claim products required for the dental care category, it is necessary for the formulator to use a variety of different ingredients . This places a number of demands on the development process . Innovations in the areas of pharmaceutical technology have contributed to the formulation of the products having superior efficacy as well as other attributes that may contribute to clinical response and patient acceptability . Improved clinical efficacy and tolerability, along with conditioning signals, should encourage patient compliance with oral hygiene further complementing professional efforts directed at disease prevention . The most effective way of preventing the development of dental disease is in controlling the production of dental plaque . It is formed by microbial action . The removal of plaque from the teeth and related areas is essential for the maintenance of a healthy mouth . In this paper we have presented the main components of toothpastes and mouthwashes . For the active ingredients, their supposed effect as therapeutic agents is also explained. Pathobiology, 2004, 71(6), 287 - 94 The periarterial space in the lung: its important role in lung edema, transplantation, and microbial or allergic inflammation; Pabst R; In mammal lungs different compartments for leukocytes can be identified during health and disease, e.g . lung interstitium, bronchoalveolar space, the epithelium and lamina propria of the air-conducting part . A so far neglected compartment is the space around the branches of the pulmonary arteries, characterized by a unique architecture of capillaries running in parallel to the pulmonary artery . This compartment - the periarterial space - is described and its physiological and pathophysiological role reviewed . The periarterial space is infiltrated by different leukocyte subsets during edema formation in the early stages of a lung transplant rejection and, in particular, during inflammatory and allergic reactions . The periarterial compartment seems to be of major relevance in all these situations . Eur Cytokine Netw, 2004 Oct-Dec, 15(4), 327 - 38 Induction of colony-stimulating factors by a 30-kDa secretory protein of Mycobacterium tuberculosis H37Rv; Kaur S et al.; Colony-stimulating factors (CSFs)-induced increased hematopoietic activity is known to occur in various microbial diseases; however, not much is known during tuberculosis (TB) . We investigated the CSF-inducing capability of a Mycobacterium tuberculosis H37Rv component . Swiss mice intravenously injected with purified 30-kDa secretory protein of M . tuberculosis H37Rv (Mtb30; 0.1-10 mg/kg) showed enhanced levels of serum CSFs; maximum response (142 +/- 16 colonies) occurred at 1 mg/kg . In vitro, Mtb30 (1-50 mug/mL) induced mouse peritoneal macrophages (PMs) to elaborate CSFs in the conditioned medium (CM); 25 mug/mL appeared optimal (97 +/- 11 colonies) . Both in vivo and in vitro, peak CSF production occurred 24 h after stimulation which levelled-off to background levels by 72 h . Rabbit anti-Mtb30 antibody significantly (p<0.05) reduced CSF production by both Mtb30-stimulated and M . tuberculosis-infected PMs, in vitro . The induced CSFs, both in the serum and CM, appeared to be functionally similar, as they supported the formation of granulocyte (G), monocyte (M) and GM colonies, in similar proportions; the GM colonies were maximum (>79 %) . Neutralizing (100%) rabbit anti-mouse interleukin-1 (IL-1) polyclonal antibody did not affect the Mtb30-induced CSF production, indicating it to be IL-1-independent; whereas, CSF production was partly dependent on tumour necrosis factor-alpha (TNF-alpha), as goat anti-mouse TNF-alpha immunoglobulin G only partly inhibited it . Mtb30-induced PM production of CSFs was de novo as it was completely blocked by cycloheximide (50 mug/mL) . The CSF-inducing capability of Mtb30 appeared to be proteinaceous in nature as it was heat (70 degrees C; 1 h)-labile, was destroyed by proteases (pronase E and trypsin) and was unaffected by sodium periodate treatment . Further, compared to the controls, Mtb30 induced significantly (p<0.05) high levels of immunoreactive GM-CSF (9+/-1 and 7.5+/-0.8 ng/mL) and M-CSF (4.3+/-0.5 and 3.9+/-0.4 ng/mL) in serum and CM, respectively; G-CSF levels did not increase significantly (p>0.05) . Mtb30-treated mice showed a maximum of 2.23- and 2.36-fold increase, in the splenic and femur colony forming unit-GM counts, respectively, as compared to the controls . This is the first report which demonstrates Mtb30-induced production of CSFs that is up-regulated both posttranscriptionally and functionally, and thus adds to our understanding of the molecular pathogenetic mechanisms of TB. Bioresour Technol, 2005 May, 96(8), 929 - 35 Bio-active composts from rice straw enriched with rock phosphate and their effect on the phosphorous nutrition and microbial community in rhizosphere of cowpea; Zayed G et al.; Composts were produced from rice straw enriched with rock phosphate and inoculated with Aspergillus niger, Trichoderma viride and/or farmyard manure (FYM) . The resulting composts were evaluated as organic phosphate fertilizers for cowpea plants in pot experiments . The results showed that the maximum amount of soluble phosphorous (1000ppm) was produced in composts inoculated with A . niger+T . viride with or without FYM . Any of the produced composts was much better than superphosphate fertilizer in providing the growing cowpea plants with phosphorous . Fertilization of the cowpea plants with the compost inoculated with FYM+A . niger+T . viride resulted in maximum amount of phosphorous uptake (295ppm) . The highest phosphate dissolving fungi numbers in rhizosphere soils of cowpea plants were obtained after fertilization with composts which received A . niger and T . viride treatments, while the highest phosphate dissolving bacterial numbers were found after fertilization with composts which received FYM treatments. Math Biosci, 2004 Dec, 192(2), 111 - 36 Epub 2004 Dec 08. Predicting stability of mixed microbial cultures from single species experiments: 2 . Physiological model; Pilyugin SS et al.; In this paper, we study the equilibria of a physiological model describing the continuous culture in which two microbial populations compete for two substitutable resources . This work is an extension of the stability analysis of the phenomenological model of mixed microbial growth {M.M . Ballyk, G.S.K . Wolkowicz, Exploitative competition in the chemostat for two perfectly substitutable resources, Math . Biosci . 118 (1993) 127-180; S.S . Pilyugin, G.T . Reeves, A . Narang, Predicting stability of mixed microbial cultures from single species experiments: 2 . Phenomenological model} . Here, we investigate the influence of the peripheral enzymes that catabolize the substrate uptake on the stability of the mixed culture . We show that, under steady state conditions, an increase in the concentration of one substrate inhibits the uptake of the other substrate(s) . We present the criteria for existence, uniqueness, and stability of various types of equilibria . We formulate these criteria in terms of growth isoclines and consumption curves for each of the competing species . Since both types of curves can be obtained from a single species experiment, our approach provides a direct connection between theory and experiment and allows one to infer the dynamics of mixed cultures from the dynamics of single species cultures . By expressing the stability criteria in terms of intracellular properties, the model establishes a link between ecology and molecular biology. Environ Health Perspect, 2005 Jan, 113(1), 6 - 10 Human colon microbiota transform polycyclic aromatic hydrocarbons to estrogenic metabolites; Van de Wiele T et al.; Ingestion is an important exposure route for polycyclic aromatic hydrocarbons (PAHs) to enter the human body . Although the formation of hazardous PAH metabolites by human biotransformation enzymes is well documented, nothing is known about the PAH transformation potency of human intestinal microbiota . Using a gastrointestinal simulator, we show that human intestinal microbiota can also bioactivate PAHs, more in particular to estrogenic metabolites . PAH compounds are not estrogenic, and indeed, stomach and small intestine digestions of 62.5 nmol naphthalene, phenanthrene, pyrene, and benzo(a)pyrene showed no estrogenic effects in the human estrogen receptor bioassay . In contrast, colon digests of these PAH compounds displayed estrogenicity, equivalent to 0.31, 2.14, 2.70, and 1.48 nmol 17alpha-ethynylestradiol (EE2), respectively . Inactivating the colon microbiota eliminated these estrogenic effects . Liquid chromatography-mass spectrometry analysis confirmed the microbial PAH transformation by the detection of PAH metabolites 1-hydroxypyrene and 7-hydroxybenzo(a)pyrene in colon digests of pyrene and benzo(a)pyrene . Furthermore, we show that colon digests of a PAH-contaminated soil (simulated ingestion dose of 5 g/day) displayed estrogenic activity equivalent to 0.58 nmol EE2, whereas stomach or small intestine digests did not . Although the matrix in which PAHs are ingested may result in lower exposure concentrations in the gut, our results imply that the PAH bioactivation potency of colon microbiota is not eliminated by the presence of soil . Moreover, because PAH toxicity is also linked to estrogenicity of the compounds, the PAH bioactivation potency of colon microbiota suggests that current risk assessment may underestimate the risk from ingested PAHs. Immunol Lett, 2005 Feb 15, 97(1), 115 - 22 Toll-like receptor 9 signaling can sensitize fibroblasts for apoptosis; Fischer SF et al.; Toll-like receptors (TLR) are activated by microbial components and transmit signals that induce cell activation and differentiation . A number of recent reports further indicate that TLR also have the potential to induce apoptosis upon ligand binding . Here we investigate the apoptosis-inducing capacity of TLR9, the receptor for microbial CpG-DNA . Unlike ligands for TLR2 and TLR4, CpG-DNA failed to induce apoptosis in RAW264.7 mouse macrophages . In human embryonic kidney fibroblasts transfected stably to express TLR9, CpG-DNA weakly induced apoptosis in one clone but not others without an obvious allocation to differences in TLR-signaling events . Analysis of the apoptotic signaling showed that the mitochondrial pathway of apoptosis was triggered by TLR9, as mitochondrial Bax was activated upstream of caspase-cleavage . CpG-DNA-induced apoptosis was reduced by cycloheximide suggesting that de novo protein synthesis was required . Strikingly, stimulation with CpG-DNA resulted in a strongly increased sensitivity of TLR9-expressing fibroblasts to apoptosis induced by staurosporine and UV-irradiation . These results identify a mitochondrial pathway to apoptosis that can be triggered by TLR9 and that may serve to sensitize cells from the innate immune system to apoptosis in the course of an immune response. J Org Chem, 2005 Jan 7, 70(1), 338 - 41 Enzyme-Catalyzed Rearrangement of a Diepoxy-germacrane Compound into New 7-epi-Eudesmane Derivatives; Garcia-Granados A et al.; Two new 7-epi-eudesmane derivatives, together with two new germacrane compounds, have been isolated from the microbial-transformation of a (1alpha,10beta),(4beta,5alpha)-diepoxygermacrane using the hydroxylating fungi Rhizopus nigricans . The rearranged skeleton and the stereochemistry of the chiral centers have been determined by means of their spectral data, and the absolute configuration has been confirmed by single-crystal X-ray analyses . A possible mechanism based on an enzyme-catalyzed isomerization to a 1alpha-hydroxy-(4beta,5alpha)-epoxygermacr-9(E)-ene intermediate and a subsequent cyclization process is proposed in order to explain the formation of the 7-epi-eudesmane compounds. Ying Yong Sheng Tai Xue Bao, 2004 Oct, 15(10), 1897 - 902 {Dynamics of microbial biomass P and its affecting factors in a long-term fertilized black soil}; Li D et al.; This paper studied the microbial biomass P in a long-term fertilized black soil . The soil was fertilized by farmyard manure (M2, M4) and chemical fertilizer (NPK), and the dynamics of soil microbial biomass P was monitored during crop growth season . The results showed that the microbial biomass P in different treatments was 8.75-47.68 mg x kg(-1) (M4), 3.02-37.16 mg x kg(-1) (M2), 1.59-10.62 mg x kg(-1) (NPK), and 0.76-6.74 mg x kg(-1) (CK) . The microbial biomass P in M4 and M2 was the highest at reproductive growth stage, and that in NPK and CK was the highest at early growth stage . The significant difference of soil microbial biomass P induced by different amounts and types of fertilizer application was not varied with the period and stage of plant growth . In all treatments, no significant correlation was found between the dynamics of soil microbial biomass P and the soil biological, physical and chemical properties in plant growth period . Soil microbial biomass P had a very significant positive correlation with soil biological, physical and chemical properties (expect K), and a significant positive correlation with the N, P and K contents of plants and soil water content. Ying Yong Sheng Tai Xue Bao, 2004 Oct, 15(10), 1891 - 6 {Dynamics of microbial biomass N in different fertilized black soil and its related factors}; Li D et al.; This paper studied the dynamics of microbial biomass N in a black soil long-term fertilized with farmyard manure (M2 and M4) and chemical fertilizer (NPK) during crop growth season . The results showed that the soil microbial biomass N was 25.52-239.12 mg x kg(-1) in M4, 10.40-94.31 mg x kg(-1) in M2, 6.27-87.04 mg x kg(-1) in NPK, and 9.15-69.81 mg x kg(-1) in CK . In same treatment, the highest value was 7-14 times higher than the lowest value . Treatments NPK and M2 had the highest value of microbial biomass N at reproductive growth stage, while M4 and CK had it at vegetative growth stage . The significant difference of soil microbial biomass N caused by different treatments was not changed with plant growth stages . The dynamics of soil microbial biomass N had a significant correlation some soil biological, physical and chemical properties . A significant positive correlation was found between soil microbial biomass N and soil biological, physical and chemical properties, plant N, P and K contents, soil moisture, and grain crude protein. Ying Yong Sheng Tai Xue Bao, 2004 Oct, 15(10), 1847 - 50 {Effects of free-air CO2 enrichment (FACE) on soil microbial biomass under rice-wheat rotation}; Li Y et al.; The effect of CO2 enrichment on soil microbial biomass in 0-5 cm and 5-10 cm soil layers under rice-wheat rotation was studied under the conditions of routine cultivation, irrigation and fertilization . The results showed that FACE treatment could significantly increase the amount of soil bacteria in 0-5 cm and 5-10 cm soil layers during rice jointing stage and wheat wintering and ripening stages . Elevated CO2 only significantly increased the amount of soil fungi in 0-5 cm soil layer during rice ripening stage . During the whole period of rotation, the amount of soil bacteria was greater than that of soil fungi. Ying Yong Sheng Tai Xue Bao, 2004 Oct, 15(10), 1842 - 6 {Soil biochemical characters of Pinus koraiensis and Pinus sylvestriformis plantations under different elevated CO2 concentration}; Jia X et al.; The aim of this study was to determine the dynamics of soil microbial biomass C, cellulose decomposition rate and catalase activity in Pinus koraiensis and Pinus sylvestriformis plantations exposed to elevated CO2 (700 mg x L(-1) and 500 mg x L(-1)) during the growth season . The five years results indicated that in Autumn, the soil microbial biomass C of Pinus koraiensis and Pinus sylvestriformis plantations was in the order of ambient CO2-chamber > 500 mg x L(-1) CO2-chamber > 700 mg x L(-1) CO2-chamber . The monthly dynamics of cellulose decomposition rate in 5 and 10 cm depth soils showed that for both Pinus sylvestriformis and Pinus koraiensis plantations, it was always higher under 500 mg x L(-1) CO2 than under 700 mg x L(-1) CO2 in growth season . Between different CO2 concentration treatments, soil catalase activity of Pinus koraiensis and Pinus sylvestriformis plantations displayed a definite pattern of its monthly dynamics. Huan Jing Ke Xue, 2004 Sep, 25(5), 122 - 6 {Diversity of microbial genes in paddy soil stressed by cadmium using DGGE}; Duan XJ et al.; Variations of diversity of microbial genes in submerged paddy soil stressed by heavy metal cadmium were studied using modern molecular biotechnology which includes directly extracting total DNA from paddy soil, amplifying 16S rDNA and their V3 variable region by PCR, the denaturing gradient gel electrophoresis (DGGE) . Two methods for extraction and purification of microbial DNA were compared . Bacterial communities were quantified by analyzing the DGGE band patterns . The genetic clusters and correlative comparison of bacterial communities were analyzed based on the DGGE finger-print . The results showed that there are some significant differences between bacterial communities in paddy soils treated with different concentrations of cadmium . The information about effect of cadium on microbial population based on molecular biological techniques are conformed with that from traditional methods, but that obtained about variations of microbial genes in paddy soil is much more than results based on the latter methods . It could provide a new way and foundation to research microbial gene diversity in contaminated environment. Isotopes Environ Health Stud, 2004 Dec, 40(4), 267 - 83 Stable isotope biogeochemistry of the sulfur cycle in modern marine sediments: I . seasonal dynamics in a temperate intertidal sandy surface sediment; Bottcher M et al.; A biogeochemical and stable isotope geochemical study was carried out in surface sediments of an organic-matter poor temperate intertidal sandy surface sediment (German Wadden Sea of the North Sea) to investigate the activity of sulfate-reducing bacteria and the dynamics of the vertical partitioning of sedimentary sulfur, iron, and manganese species in relation to the availability of total organic carbon (TOC) and mud contents . The contents and stable isotopic compositions ((34)S/(32)S) of total reduced inorganic sulfur species (TRIS) and dissolved sulfate were measured . Maximum oxygen penetration depths were estimated from the onset of a blackening of the sediments due to FeS accumulation and ranged from 5 to 10 mm below surface (mmbsf) . A zone of relatively moderate relative organic-matter enrichment was found between 5 and 20 mmbsf leading to enhanced activities of sulfate-reducing bacteria with sulfate-reduction rates (SRR) up to 350 nmol cm(-3) d(-1) . Below this zone, microbial SRR dropped significantly . Depth integrated SRR seem to depend not only on temperature but also on the availability of reactive organic matter . The sulfur-isotopic composition of TRIS was depleted in (34)S by 33-40 per thousand with respect to coexisting dissolved sulfate (constant at about +21 per thousand vs . Vienna-Canyon Diablo Troilite (V-CDT)) . Since sulfate reduction is not limited by dissolved sulfate (open system), depth variations of the isotopic composition of TRIS reflect changes in overall isotope effect due to superimposed microbial and abiotic reactions . Most of the solid-phase iron and manganese was bonded to (non-reactive) heavy minerals . However, a layer of reactive Fe(III) and Mn(IV) oxi(hydroxi)des was found in the uppermost sediment section due to re-oxidation of dissolved Fe(II) and Mn(II) species at the sediment-water interface . Metal cycling below the surface is at least partially coupled to intense sulfur cycling. Free Radic Res, 2004 Nov, 38(11), 1143 - 53 Effect of methylguanidine in a model of septic shock induced by LPS; Marzocco S et al.; Septic shock, a severe form of sepsis, is characterized by cardiovascular collapse following microbial invasion of the body . The progressive hypotension, hyporeactivity to vasopressor agents and vascular leak leads to circulatory failure with multiple organ dysfunction and death . Many inflammatory mediators (e.g . TNF-alpha, IL-1 and IL-6) are involved in the pathogenesis of shock and, among them, nitric oxide (NO) . The overproduction of NO during septic shock has been demonstrated to contribute to circulatory failure, myocardial dysfunction, organ injury and multiple organ failure . We have previously demonstrated with in vitro and in vivo studies that methylguanidine (MG), a guanidine compound deriving from protein catabolism, significantly inhibits iNOS activity, TNF-alpha release and carrageenan-induced acute inflammation in rats . The aim of the present study was to evaluate the possible anti-inflammatory activity of MG in a model of septic shock induced by lipopolysaccharide (LPS) in mice . MG was administered intraperitoneally (i.p.) at the dose of 30 mg/kg 1 h before and at 1 and 6 h after LPS-induced shock . LPS injection (10 mg/kg in 0.9% NaCl; 0.1 ml/mouse; i.p.) in mouse developed a shock syndrome with enhanced NO release and liver, kidney and pancreatic damage 18 h later . NOx levels, evaluated as nitrite/nitrate serum levels, was significantly reduced in MG-treated rats (78.6%, p < 0.0001; n = 10) . Immunohistochemistry revealed, in the lung tissue of LPS-treated group, a positive staining for nitrotyrosine and poly(adenosine diphosphate {ADP} ribose) synthase, both of which were reduced in MG-treated mice . Furthermore, enzymatic evaluation revealed a significant reduction in liver, renal and pancreatic tissue damage and MG treatment also improved significantly the survival rate . This study provides evidence that MG attenuates the degree of inflammation and tissue damage associated with endotoxic shock in mice . The mechanisms of the anti-inflammatory effect of MG is, at least in part, dependent on the inhibition of NO formation. J Asian Nat Prod Res, 2005 Sep, 7(3), 225 - 9 Microbial hydroxylation of cinobufagin by Mucor spinosus; Zhang W et al.; Mucor spinosus has been employed for the biotransformation of cinobufagin (1) to afford three metabolites . On the basis of their physico-chemical data, the structures of the transformation products have been characterized as 1beta-hydroxy-cinobufagin (2), 12beta-hydroxy-cinobufagin (3) and 1beta,12beta-dihydroxy-cinobufagin (4), of which metabolites 2 and 4 are new compounds . In vitro cytotoxic activities of the biotransformation products and the substrate-cinobufagin have been assayed against four tumor cell lines of Bel 7420, BGC 823, HeLa and HL 60; they all showed cytotoxic activities. FEMS Microbiol Lett, 2005 Jan 1, 242(1), 37 - 44 Incomplete processing of peroxidase transcripts in the lignin degrading fungus Phanerochaete chrysosporium; Macarena S et al.; Phanerochaete chrysosporium has been thoroughly studied as a microbial model for lignin degradation . The enzymes lignin peroxidase (LiP) and manganese peroxidase (MnP), both encoded by several genes, play the main role in the cleavage of different lignin substructures . In this work, the expression of specific LiP and MnP transcripts in liquid medium and in a wood-containing soil system was studied by reverse transcription-PCR and subsequent cloning and sequencing of the products obtained . Splice variants of different LiP and MnP transcripts were observed in wood-containing soil incubations and in liquid cultures . The processed transcripts contained different numbers of complete introns . Since the presence of stop codons in several of these introns would prevent the synthesis of active enzyme, we propose that these transcripts arise as a result of incomplete processing rather than alternative splicing . Interestingly, analysis of splice variants from mnp genes led to the identification of a fourth actively transcribed gene coding for MnP in P . chrysosporium. Environ Pollut, 2005 Apr, 134(3), 535 - 48 Practical considerations for addressing uncertainties in monitoring bulk deposition; Dammgen U et al.; The assessment of the deposition of both wet (rain and cloud) and dry sedimenting particles is a prerequisite for estimating element fluxes in ecosystem research . Many nations and institutions operate deposition networks using different types of sampler . However, these samplers have rarely been characterized with respect to their sink properties . Major errors in assessing bulk deposition can result from poor sampling properties and defective sampling strategies . Relevant properties are: sampler geometry and material, in particular the shape of the rim; sink properties for gases and aerosols; and microbial transformations of the collected samples . An adequate number of replicates allows the identification of samples which are contaminated, in particular by bird droppings . The paper discusses physical and chemical properties of the samplers themselves . The dependence of measurement accuracy on the number of replicates and the sampling area exposed is discussed . Recommendations are given for sampling strategies, and for making corrections and substitution of missing data. J Hosp Infect, 2005 Feb, 59(2), 90 - 5 Antibiotic prescription practice in an intensive care unit using twice-weekly collection of screening specimens: a prospective audit in a large UK teaching hospital; Warren MM et al.; Controversy exists regarding the optimal method of managing intensive care unit (ICU)-acquired infection . Antibiotic prescriptions in 177 sequential admissions to our ICU, which collected twice-weekly screening samples, were audited . Seventy-seven percent of patients received at least one antibiotic prescription, and 45% of patients received at least one prescription for suspected or proven sepsis . Of the 353 antibiotic prescriptions audited, 86 were prophylactic and 61 were first prescribed prior to ICU admission . One hundred and eighty-three were prescribed for sepsis; of these, 108 (59%) were empirical prescriptions and only 21% of these were subsequently changed . For the 75 prescriptions for specific organisms, 28% targeted organisms isolated at least four days previously . Clinicians in our ICU reviewed the data and reached consensus that screening was associated with decision making that did not represent current evidence-based practice, because empirical prescriptions were rarely changed or stopped on the basis of new samples, and those prescribed for confirmed infection frequently targeted organisms isolated before the septic episode . After our audit, we stopped regular collection of screening samples and used more targeted and invasive sampling, in response to clinical suspicion, to guide therapy and maintain data concerning local microbial epidemiology. J Nat Prod, 2004 Dec, 67(12), 2133 - 5 Natural Aromatic Steroids as Potential Molecular Fossils from the Fruiting Bodies of the Ascomycete Daldinia concentrica; Qin XD et al.; The identification of aromatic steroid hydrocarbons bearing a methyl group at positions 1, 2, 3, 4, or 6 in sediments and petroleum has been puzzling since possible steroidal precursors have not yet been reported in living organisms . Two new aromatic steroids, (17beta,20R,22E,24R)-19-norergosta-1,3,5,7,9,14,22-heptaene (1) and (17beta,20R,22E,24R)-1-methyl-19-norergosta-1,3,5,7,9,14,22-heptaene (2), were isolated from the ascomycete Daldinia concentrica, of which compound 2 bears an unusual methyl group at position 1 . We propose that the origin of these compounds is derived from the transformation undergone by their precursor due to microbial action . Compounds 1 and 2 could be the long-sought, biological precursor steroids for organic matter in Earth's subsurface. Science, 2004 Dec 24, 306(5705), 2216 - 21 Distributions of microbial activities in deep subseafloor sediments; D'Hondt S et al.; Diverse microbial communities and numerous energy-yielding activities occur in deeply buried sediments of the eastern Pacific Ocean . Distributions of metabolic activities often deviate from the standard model . Rates of activities, cell concentrations, and populations of cultured bacteria vary consistently from one subseafloor environment to another . Net rates of major activities principally rely on electron acceptors and electron donors from the photosynthetic surface world . At open-ocean sites, nitrate and oxygen are supplied to the deepest sedimentary communities through the underlying basaltic aquifer . In turn, these sedimentary communities may supply dissolved electron donors and nutrients to the underlying crustal biosphere. J Biol Chem . 2004 Dec 23; {Epub ahead of print} Discovery of pectin-degrading enzymes and directed evolution of a novel pectat lyase for processing cotton fabric; Solbak AI et al.; There is a growing need in the textile industry for more economical and environmentally responsible approaches to improve the scouring process as part of the pretreatment of cotton fabric . Enzymatic methods using pectin-degrading enzymes are potentially valuable candidates in this effort because they could reduce the amount of toxic alkaline chemicals currently used . Using high throughput screening of complex environmental DNA libraries more than 40 novel microbial pectate lyases were discovered, and their enzymatic properties were characterized . Several candidate enzymes were found that possessed pH optima and specific activities on pectic material in cotton fibers compatible with their use in the scouring process, however, none exhibited the desired temperature characteristics . Therefore, a candidate enzyme was selected for evolution . Using Gene Site Saturation Mutagenesis (GSSMTM) technology, thirty-six single site mutants exhibiting improved thermotolerance were produced . A combinatorial library derived from the twelve best performing single site mutants was then generated by using Gene ReassemblyTM technology . Nineteen variants with further improved thermotolerance were produced . These variants were tested for both improved thermotolerance and for performance in the bioscouring application . The best performing variant (CO14) contained 8 mutations and had a melting temperature 16oC higher than the wild type enzyme while retaining the same specific activity at 50oC . Optimal temperature of the evolved enzyme was 70oC which is 20oC higher than the wild type . Scouring results obtained with the evolved enzyme were significantly better that the results obtained with chemical scouring, making it possible to replace the conventional and environmentally harmful chemical scouring process. Nature . 2004 Dec 22; {Epub ahead of print} Plastid proteins crucial for symbiotic fungal and bacterial entry into plant roots; Imaizumi-Anraku H et al.; The roots of most higher plants form arbuscular mycorrhiza, an ancient, phosphate-acquiring symbiosis with fungi, whereas only four related plant orders are able to engage in the evolutionary younger nitrogen-fixing root-nodule symbiosis with bacteria . Plant symbioses with bacteria and fungi require a set of common signal transduction components that redirect root cell development . Here we present two highly homologous genes from Lotus japonicus, CASTOR and POLLUX, that are indispensable for microbial admission into plant cells and act upstream of intracellular calcium spiking, one of the earliest plant responses to symbiotic stimulation . Surprisingly, both twin proteins are localized in the plastids of root cells, indicating a previously unrecognized role of this ancient endosymbiont in controlling intracellular symbioses that evolved more recently. J Dent Res, 2005 Jan, 84(1), 59 - 63 Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis are Components of a Polymicrobial Intracellular Flora within Human Buccal Cells; Rudney JD et al.; Previously, we used in situ hybridization and confocal microscopy to detect the periodontal pathogens Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis within buccal epithelial cells taken directly from the mouth . This study tested the hypothesis that the intracellular flora of buccal cells is polymicrobial . Mixtures containing a red fluorescent universal probe paired with green fluorescent versions of either A . actinomycetemcomitans-, P . gingivalis-, or T . forsythensis-specific probes were hybridized with buccal cells collected from each of 38 healthy humans . We verified co-localization of probe pairs within cells by generating three-dimensional reconstructions . Intracellular bacteria were detected in every subject . Each cell that was labeled with a species-specific probe also contained bacteria recognized only by the universal probe . Bacteria labeled with specific probes often occupied smaller regions within larger masses of bacteria . Those findings suggest that future studies of invasion by oral bacteria may need to include microbial consortia. Appl Microbiol Biotechnol . 2004 Dec 22; {Epub ahead of print} Streptomycete spores entrapped in chitosan beads as a novel biocontrol tool against common scab of potato; Jobin G et al.; Spores of Streptomyces melanosporofaciens EF-76, an actinomycete that inhibits the growth of several plant pathogens, were incorporated in beads of chitosan and polyphosphate using the entrapment technique called complex coacervation . The degradation of spore-loaded beads was monitored by measuring the residual amount of chitosan in soil and by enumerating the S . melanosporofaciens population over time . After the introduction of spore-loaded chitosan beads into soil, the amount of chitosan in sterile soil remained at 1.550 mg/g throughout the first week and diminished to 0.101 mg/g after 7 weeks . Bead degradation proceeded faster in non-sterile soil but a progressive release of both chitosan oligomers and the antagonistic microbial agent was nevertheless observed . Application of these spore-loaded chitosan beads to seed potato tubers protected progeny tubers against common scab. Microb Ecol . 2004 Dec 21; {Epub ahead of print} Biogeochemistry of an Iron-Rich Hypersaline Microbial Mat (Camargue, France); Wieland A et al.; In situ microsensor measurements were combined with biogeochemical methods to determine oxygen, sulfur, and carbon cycling in microbi