Microbial

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–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 microbial mats growing in a solar saltern (Salin-de-Giraud, France) . Sulfate reduction rates closely followed the daily temperature changes and were highest during the day at 25 degrees C and lowest during the night at 11 degrees C, most probably fueled by direct substrate interactions between cyanobacteria and sulfate-reducing bacteria . Sulfate reduction was the major mineralization process during the night and the contribution of aerobic respiration to nighttime DIC production decreased . This decrease of aerobic respiration led to an increasing contribution of sulfide (and iron) oxidation to nighttime O(2) consumption . A peak of elemental sulfur in a layer of high sulfate reduction at low sulfide concentration underneath the oxic zone indicated anoxygenic photosynthesis and/or sulfide oxidation by iron, which strongly contributed to sulfide consumption . We found a significant internal carbon cycling in the mat, and sulfate reduction directly supplied DIC for photosynthesis . The mats were characterized by a high iron content of 56 mumol Fe cm(-3), and iron cycling strongly controlled the sulfur cycle in the mat . This included sulfide precipitation resulting in high FeS contents with depth, and reactions of iron oxides with sulfide, especially after sunset, leading to a pronounced gap between oxygen and sulfide gradients and an unusual persistence of a pH peak in the uppermost mat layer until midnight.

J Immunother, 2005 Jan-Feb, 28(1), 73 - 8
Human gammadelta T cells from G-CSF-mobilized donors retain strong tumoricidal activity and produce immunomodulatory cytokines after clinical-scale isolation; Otto M et al.; Human gammadelta T cells are a small fraction of T cells that have been shown to exert major histocompatibility (MHC)-unrestricted natural cytotoxicity against a variety of solid tumors and some subsets of leukemias and lymphomas . They are also involved in the immune response to certain bacterial, viral, and parasitic infections and expand significantly in CMV- or HSV-infected organ allografts . They are able to mediate antibody-dependent cytotoxicity and are not alloreactive, which makes them attractive candidates for cell-based immunotherapy . However, their frequency in peripheral blood is low and ex vivo expansion of gammadelta T cells is labor-extensive, does not always yield cells with full innate cytotoxic power, and has the potential for microbial contamination . Therefore, the authors developed a clinical-scale, automated cell purification method for the efficient enrichment of gammadelta T cells from leukapheresis products . Six leukapheresis products were purified for gammadelta T cells using a single-step immunomagnetic method . Purity and phenotype were assessed by flow cytometry . A standard Europium release assay was performed to determine the cytotoxic capacity of the cells . Cytokine production was measured using a multiplex sandwich immunoassay . The mean percentage of gammadelta T cells in the final product was 91%, with an average recovery of 63% . The cells showed a high co-expression of CD8, CD56, CD28, and CD11b/CD18 . In some products an unusually high proportion of Vgamma9Vdelta1 T cells was found . The isolated cells were cytotoxic against the neuroblastoma cell line NB1691 and the erythroleukemic line K562 in vitro . They were able to produce a variety of immunomodulatory cytokines such as IFNgamma, TNFalpha, and MIP-1beta, but also GM-CSF and G-CSF when co-incubated in culture with and without various stimuli . In summary, the authors describe a rapid, automated, and efficient method for the large-scale enrichment of human gammadelta T cells . The cytotoxic properties of the cells were preserved . This method yields sufficient purified gammadelta T cells for use in adoptive immunotherapy as well as laboratory investigations and animal studies.

J Vet Sci, 2004 Dec, 5(4), 337 - 43
Development of a novel antigen capture-ELISA using IgY against porcine interleukin-6 and its application; Lee DY et al.; Interleukin-6 (IL-6) is introduced as a marker of disease . At present, a variety of method may be used to quantify expression of this protein . Antigen capture-ELISA is a sensitive and accurate quantification method previously used with ovine, rat, and human IL-6 proteins . However, it has never been reported to quantify porcine IL-6 protein using capture ELISA . In this study, we generated and characterized a set of IgY and mono-specific polyclonal antibodies to recombinant porcine IL-6 (rpIL-6), and combining these with a sensitive and specific capture-ELISA for a diagnostic purpose . cDNA encoding the mature protein coding region of porcine IL-6 was cloned and expressed with pQE-30UA expression vector . rpIL-6 was then expressed and purified by using Ni-NTA resin . Protein mass of 24 kDa was found with SDS-PAGE and the identity of the protein was confirmed by Western-blot . Production of polyclonal antibodies against rpIL-6 was performed using the purified rpIL-6 in mice and hens . An antigen capture-ELISA was developed with the antibodies after their extraction . To compare the IL-6 level in the different sanitary state of farms, pig sera were randomly collected and concentration of IL-6 in the sera was measured with the antigen capture-ELISA . The capture-ELISA with the optimal concentration of antibodies, in this study, was able to detect about 10 ng/ml of rpIL-6 . IL-6 levels determined with the capture-ELISA in pig sera showed positive correlation with the sanitary states of the farms . These results suggested that the developed antigen capture-ELISA could be a good tool for the screening of microbial infection in pig farms.

J Agric Food Chem, 2004 Dec 29, 52(26), 7809 - 13
Rapid fluorescence screening assay for enrofloxacin and tetracyclines in chicken muscle; Schneider MJ; A simple, rapid fluorescence assay was developed for screening both enrofloxacin (ENRO) and tetracyclines in chicken muscle at the U.S . tolerance levels (300 ng/g and 2 microg/g, respectively) . Screening for both classes of antibiotics is accomplished using one extraction, thus simplifying and expediting the process . The method requires an initial extraction of chicken muscle with 1% acetic acid in acetonitrile, centrifugation, and analysis of the supernatant for ENRO fluorescence . After addition of ammonium hydroxide, magnesium chloride, and methanol, followed by centrifugation and filtration, the supernatant can be measured for tetracycline fluorescence . Chlortetracycline (CTC) was chosen as a representative tetracycline to demonstrate the method, as it displays intermediate sensitivity among the three tetracyclines approved in the U.S . Comparison of the fluorescence of control and tolerance-level-fortified samples of both ENRO and CTC shows no overlap . Setting a threshold as the average fortified fluorescence minus 3sigma allows for successful screening, as illustrated with blind samples as controls or fortified with ENRO and/or CTC over a range of concentrations . This method can provide an alternative or supplemental approach to currently used microbial screening assays.

J Immunol, 2005 Jan 1, 174(1), 27 - 30
Cutting Edge: NF-kappaB-activating kinase-associated protein 1 participates in TLR3/Toll-IL-1 homology domain-containing adapter molecule-1-mediated IFN regulatory factor 3 activation; Sasai M et al.; TLRs signal the presence of microbial patterns and activate transcription factors . In TLR3 and TLR4, the adapter Toll-IL-1R homology domain-containing adapter molecule (TICAM-1) (also called Toll/IL-1R domain-containing adapter inducing IFN-beta (TRIF)) mediates IFN regulatory factor 3 (IRF3) phosphorylation followed by IFN-beta production . The regulatory subunit TNFR-associated factor family member-associated NF-kappaB activator (TANK) couples with the kinase complex IkappaB kinase-related kinase epsilon/NF-kappaB-activating kinase (NAK) (TANK-binding kinase 1 (TBK1)) that involveTICAM-1-dependent IFN-beta induction . There are several TANK-homologous proteins . We tested whether TICAM-1 binds and coprecipitates with TANK or its family proteins . The results are: 1) the TANK family protein NAK-associated protein 1 (NAP1), but not TANK, coprecipitates withTICAM-1; 2) NAP1 overexpression markedly enhances TBK1-mediated IFN-beta promoter activation; 3) a dominant-negative form, NAP (158-270), suppresses IRF3 activation in response to poly(I:C) or LPS; 4) RNA interference targeting of the NAP1 message results in a failure of poly(I:C)-mediated IRF3 polymerization and IFN-beta production . Thus, NAP1 is the kinase subunit responsible for TLR3/4-mediated IFN-beta induction in the TICAM-1 pathway.

Chemotherapy, 2004 Dec, 50(6), 283 - 8 Epub 2004 Dec 08.
Effect of fludarabine on leukocyte functions; Dogan AL et al.; BACKGROUND: Fludarabine induces leukemic cell apoptosis and is highly efficient in chronic lymphocytic leukemia . However, fludarabine therapy causes severe leukopenia . Leukocyte myeloperoxidase (MPO) catalyzes the formation of HOCl, and this is the main microbicidal function in phagocytes . The aim of our study was to evaluate the effect of fludarabine on leukocytes, i.e . their degranulation capacity, MPO activity and HOCl production . METHODS: Peripheral blood leukocytes were incubated for 48 h with fludarabine . Degranulation was measured using a flow-cytometric method . MPO activity and HOCl production were measured spectrophotometrically . RESULTS: The degranulation capacity of fludarabine-treated leukocytes was significantly elevated compared to untreated controls . MPO activity and HOCl production were also increased in parallel . A possible direct activating effect of fludarabine was tested on the MPO activity of HL60 cells . Fludarabine did not affect MPO activity at concentrations ranging from 10 microM to 2 mM . CONCLUSION: Fludarabine had no inhibitory effect on the microbial killing of leukocytes . 2004 S . Karger AG, Basel.

Appl Radiat Isot, 2005 Mar, 62(3), 389 - 93
Preparation and in vitro evaluation of radioiodinated bakuchiol as an anti tumor agent; Bapat K et al.; Bakuchiol, extracted from the plant Psoralea corylifolia, has been proven to have anti-tumor, cytotoxic, anti-microbial and anti-inflammatory activity . In order to study if radiolabeled bakuchiol exhibits enhanced cytotoxicity, bakuchiol was radiolabeled with (125)I . In-vitro uptake studies of (125)I-bakuchiol were carried out using LS-A (lymphosarcoma) and barcl-95 (radiation-induced thymic lymphoma) ascitic and solid tumor cells of murine origin . In both LS-A and barcl-95, (125)I-bakuchiol showed significant uptake . Viability studies showed that the radioiodinated compound showed greater cytotoxic effect than bakuchiol.

Mol Immunol, 2005 Feb, 42(4), 445 - 50
Mouse Ly49 NK receptors: balancing activation and inhibition; Ortaldo JR et al.; Previous studies from numerous laboratories have demonstrated that inhibitory class I binding NK receptors dominate functional interactions in vitro . Our previous studies have shown that in addition to lysis, a major consequence of triggering the murine activating NK receptor Ly49D is the expression of cytokines and chemokines . We have recently shown that the activating Ly49D murine NK cell receptor can potently synergize during co-stimulation with IL-12 and IL-18 for selective production of IFN-gamma . Activation both in vitro and in vivo and synergistic production of IFN-gamma by Ly49D expressing NK cells results from cytokine stimulation combined with co-receptor ligation . In addition, IL-12 is capable of overriding the inhibitory receptor blockade for cytokine production, both in vitro and in vivo . Our current studies will expand this finding of IL-12 synergy to other receptors in the NK repertoire and evaluate potential biochemical mechanisms involved in this synergy . These findings place NK cells and their activating Ly49 receptors as important initiators of microbial, antiviral and anti-tumor immunity and provide a mechanism for the release of activating Ly49 receptors from an inhibitory receptor blockade . Discussion of how activation of the innate immune system provides important initiators of adaptive immune responses by receptor cross-linking and cytokine co-receptor engagement will ensue.

J Chromatogr B Analyt Technol Biomed Life Sci, 2005 Jan 5, 814(1), 133 - 41
LC-MS/MS determination in rabbit plasma of the main photoproduct of RLP068/Cl, a cationic sensitizer proposed for photodynamic therapy (PDT) of microbial infections; Chiti G et al.; The clinical development of a sensitizer for photodynamic therapy (PDT) requires the structural identification of the photoproducts and their quantification in biological fluids and tissues . We describe the LC-MS identification of the most important photoproducts of a cationic phthalocyanine sensitizer (RLP068/Cl) and a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of the main photoproduct (the cationic phthalimide derivative 3-{(1,3-dioxo-2,3-dihydro-1H-isoindol-4-yl)oxy}-N,N,N-trimethylbenzenaminium chloride) in rabbit plasma . The tri-deuterated product was used as co-eluting internal standard . The cationic photoproduct was isolated from plasma samples by protein precipitation with perchloric acid in methanol (7%, v/v) . HPLC step was performed on a Phenomenex Synergi Hydro-RP column (20mmx2.0mm, 2mum particles) with a mobile phase of 0.5% (v/v) aqueous TFA/methanol (85:15, v/v) . Flow rate was 0.2mL/min and 40muL injection were performed . Run time was 10min . Detection was achieved by means of a Bruker Esquire 3000+ ion trap mass spectrometer equipped with an ESI source working in positive mode . A multiple reaction monitoring method following the transitions 297.1 --> 282.1 for the analyte and 300.1 --> 282.1+285.1 for the internal standard was used . The analytical method was validated over the concentration range 0.46-91.2ng/mL and lower limits of detection (LLOD) and quantification (LLOQ) respectively of 0.2 and 0.5ng/mL were found.

Health Policy, 2005 Feb, 71(2), 133 - 49
Regulatory control of Chinese Proprietary Medicines in Singapore; Yee SK et al.; In Singapore, there has been a growing public interest in traditional Chinese medicine (TCM), which plays an important role in the healthcare system . With effect from 1 September 1999, the control on Chinese Proprietary Medicines (CPM) was implemented in three phases over a span of 3 years . Under the CPM regulatory framework, CPM importers, wholesalers, manufacturers and re-packers must be licensed and CPM products assessed and listed by the health authority before they are allowed for sale . Product assessment is based on the criteria of safety and quality which include limits of toxic heavy metals and microbial contents . During the 3-year CPM listing exercise, irregularities were detected, including intrinsic toxicity, adulteration, substitution, contamination, misidentification, non-traditional and/or inappropriate usage, incorrect preparation, inappropriate labelling, exaggerated claims and/or advertising . The full implementation of CPM control by September 2001 saw the listing of 7943 CPM products and brought about an improvement in the quality and safety of CPM in Singapore as well as the removal of sub-standard, fake/imitation and adulterated products . The standards of practices by local CPM dealers have also been raised . These improvements have, in turn, increased the level of public confidence in CPM products in Singapore.

Bioresour Technol, 2005 May, 96(7), 785 - 790
Hydrolytic enzyme activities of extracted humic substances during the vermicomposting of a lignocellulosic olive waste; Benitez E et al.; Humic substances and three hydrolytic enzymes (beta-glucosidase, phosphatase and urease) were extracted by neutral sodium pyrophosphate from an olive waste (dry olive cake), alone or mixed with municipal biosolids, during a nine month vermicomposting process . Easily degradable compounds decreased during the vermicomposting process because of microbial consumption . When municipal biosolids were added to dry olive cake, microbial activity increased and the amounts of compounds extracted by pyrophosphate were three times lower than olive cake alone . In both instances, beta-glucosidase, phosphatase and urease activities of the organic extracts either increased or remained the same after a nine month period of vermicomposting, thus suggesting that the humus enzyme complexes resisted microbial and earthworm attack . It is known that humus immobilised enzymes also remain active in soil environments, reactivating the nutrient cycles in soil . The use as amendments of vermicomposted olive cake, alone or when mixed with biosolids, could be a good alternative to reactivate the C, P and N-cycles in degraded soils for regeneration purposes.

Immunology, 2005 Jan, 114(1), 83 - 93
Deviation from major codons in the Toll-like receptor genes is associated with low Toll-like receptor expression; Zhong F et al.; Microbial structures activate Toll-like receptors (TLRs) and TLR-mediated cell signalling elicits and regulates host immunity . Most TLRs are poorly expressed but the underlying expression mechanism is not clear . Examination TLR sequences revealed that most human TLR genes deviated from using major human codons . CD14 resembles TLRs in sequence but its gene preferentially uses major codons . Indeed, CD14 expression on monocytes was higher than expression of TLR1 and TLR2 . The TLR9 gene is abundant in major codons and it also showed higher expression than TLR1, TLR2 and TLR7 in transfected 293T cells . Change of the 5'-end 302 base pairs of the TLR2 sequence into major human codons markedly increased TLR2 expression, which led to increased TLR2-mediated constitutive nuclear factor-kappaB activation . Change of the 5'-end 381 base pairs of the CD14 sequence into prevalent TLR codons markedly reduced CD14 expression . These results collectively show that the deviation of TLR sequences from using major codons dictates the low TLR expression and this may protect the host against excessive inflammation and tissue damages.

Eur J Biochem, 2004 Dec, 271(23-24), 4606 - 12
Nucleocytoplasmic shuttling of STAT transcription factors; Meyer T et al.; The signal transducer and activator of transcription (STAT) proteins have initially been described as cytoplasmic proteins that enter the nucleus only after cytokine treatment of cells . Contrary to this assumption, it was demonstrated that STATs are constantly shuttling between nucleus and cytoplasm irrespective of cytokine stimulation . This happens both via carrier-dependent as well as carrier-independent transportation . Moreover, it was also recognized that cytokine stimulation triggers nuclear retention of dimeric STATs, rather than affecting the rate of nuclear import . In summary, it is increasingly being appreciated that STAT nucleocytoplasmic cycling determines the quality of cytokine signaling and also constitutes an important area for microbial intervention.

Aviakosm Ekolog Med, 2004 Sep-Oct, 38(5), 46 - 52
{Air purification aboard space vehicles}; Quality of milk and of Canestrato Pugliese cheese from ewes exposed to different ventilation regimens; Dipartimento PRIME Universita di Foggia, ItalyEffects of ventilation regimen on the quality of ewes' milk and on proteolysis in Canestrato Pugliese cheese during ripening were studied . Cheeses were manufactured from the bulk milk of Comisana ewes subjected to three different ventilation regimens, which were designated low (LOV, 23 m3/h per ewe), moderate (MOV, 47 m3/h per ewe) and programmed ventilation regimen (PROV, 73 m3/h per ewe; fan set to maintain 70% relative humidity) . Bulk milk was analysed for chemical and microbial composition, renneting parameters and plasmin-plasminogen activities . At 1, 15, 30 and 45 d of ripening, the cheeses were analysed for gross chemical composition, nitrogen fractions, and plasmin and plasminogen activities . The pH 4.6-insoluble nitrogen fractions were analysed by urea-PAGE . Free amino acid content was determined at the end of ripening . Lower concentrations of bulk milk somatic cell count (BMSCC) and of mesophilic bacteria were found in the MOV group than in the LOV and the PROV groups . A lower plasminogen (PG) to plasmin (PL) ratio (PG/PL) was observed in the MOV and PROV than in the LOV cheeses . Irrespective of treatment, PL activity in cheeses was higher at 15d of ripening, while a sudden decrease of PL and PG activities was observed at 30 d, which was associated with a marked increase in non-protein nitrogen . The peptide profile characterized in the urea-PAGE showed a greater intensity of alpha- and beta-CN hydrolysis in the MOV than in the PROV and LOV cheeses . The results provide evidence that a proper ventilation regimen is critical for optimizing the hygienic quality of milk and the proteolysis of Canestrato Pugliese cheese during ripening.

Biotechnol Lett, 2004 Nov, 26(22), 1717 - 21
Improving the dynamic response of a mediator-less microbial fuel cell as a biochemical oxygen demand (BOD) sensor; Moon H et al.; The dynamic behavior of a mediator-less, microbial fuel cell (MFC) was studied as a continuous biochemical oxygen demand (BOD) sensor . The response time and the sensitivity were analyzed through the step-change testing of the fuel concentration . The MFC of 25 ml had the shortest response time of 36+/- 2 min at the fuel-feeding rate of 0.53 ml min(-1) and the resistance of 10Omega A smaller MFC of 5 ml had a response time of 5+/- 1 min.

J Environ Radioact, 2005, 79(2), 107 - 18
Radiocesium storage in soil microbial biomass of undisturbed alpine meadow soils and its relation to (137)Cs soil-plant transfer; Stemmer M et al.; This study focuses on radiocesium storage in soil microbial biomass of undisturbed alpine meadow sites and its relation to the soil-to-plant transfer . Soil and plant samples were taken in August 1999 from an altitude transect (800-1600m.a.s.l.) at Gastein valley, Austria . Soil samples were subdivided into 3-cm layers for analyses of total, K(2)SO(4)-extractable and microbially stored (137)Cs . Microbial biomass was measured by the fumigation extraction method, and fungal biomass was quantified using ergosterol as biomarker molecule . In general, the quantity of (137)Cs stored in the living soil microbial biomass was relatively small . At the high-altitude meadows, showing high amounts of fungal biomass, microbially stored (137)Cs amounted to 0.64+/-0.14kBqm(-2) which corresponds to about 1.2-2.7% of the total (137)Cs soil inventory . At lower altitudes, microbial (137)Cs content was distinctly smaller and in most cases not measurable at all using the fumigation extraction method . However, a positive correlation between the observed soil-to-plant aggregated transfer factor, microbially stored (137)Cs and fungal biomass was found, which indicates a possible role of fungal biomass in the storage and turnover of (137)Cs in soils and in the (137)Cs uptake by plants.

Mar Environ Res, 2005 Jun, 59(5), 493 - 518
Field data analysis and application of a complex water column biogeochemical model in different areas of a semi-enclosed basin: towards the development of an ecosystem management tool; Petihakis G et al.; The Pagasitikos gulf ecosystem is studied through the analysis of experimental field data acquired during several monitoring projects and the application of a complex biogeochemical model . The gulf was separated into three different parts (internal, top central-external, bottom central-external) according to the patterns exhibited by the key ecosystem indicators . Unlike other semi-enclosed gulfs Pagasitikos can be characterised as meso-oligotrophic undergoing periods of P or N limitation . Although the signal of nutrient inputs is not very clear in the field data, their importance is assessed through simulation . Increased phosphate concentrations either due to mixing or due to anthropogenic activities can result in phytoplanktonic blooms with significant contribution by diatoms . The effect of hydrodynamic patterns on primary production has been demonstrated through ecosystem modeling indicating that due to long stratification periods, all nutrients released through the benthic regeneration are trapped in the deeper layers, developing a microbial food web . However when the thermocline erodes nutrients find their way up in the upper layers of the euphotic zone and the system turns into more classical type with primary producers growing significantly faster.

Biochem J . 2004 Dec 16; {Epub ahead of print}
Epolactaene binds human Hsp60 Cys 442 resulting in the inhibition of chaperone activity; Nagumo Y et al.; Epolactaene is a microbial metabolite isolated from Penicillium sp., from which we synthesized its derivative epolactaene tertiary butyl ester (ETB) . Here, we report on the identification of the binding proteins of epolactaene/ETB and the results of our investigation into its inhibitory mechanism . Using biotin-labeled derivatives of epolactaene/ETB, human heat shock protein 60 (Hsp60) was identified as a binding protein of epolactaene/ETB in vitro as well as in situ . In addition, we found that Hsp60 preincubated with epolactaene/ETB lost its chaperone activity . The in vitro binding study showed that biotin-conjugated epolactaene/ETB covalently binds to Hsp60 . In order to investigate the binding site, binding experiments with Ala mutants of Hsp60 cysteins were conducted . As a result, it was suggested that Cys 442 is responsible for the covalent binding with biotin-conjugated epolactaene/ETB . Furthermore, the replacement of Hsp60 Cys 442 with an alanine renders the chaperone activity resistant to ETB inhibition, while the alanine replacement of other cysteins do not . These results indicate that this cysteine residue is alkylated by ETB, leading to Hsp60 inactivation.

Curr Opin Drug Discov Devel, 2004 Mar, 7(2), 204 - 10
Toll-like receptors involved in the response to microbial pathogens: development of agonists for toll-like receptor 9; Schetter C et al.; Upon microbial infection, a host has to mount a multiplicity of immune responses that target the invading pathogen . This is achieved in part by the use of particular receptors expressed on mammalian innate immune cells, the toll-like receptors (TLRs) . Recognition of a given microbial molecular structure, such as bacterial or viral DNA, leads to the activation of signaling pathways that result in distinct sets of immune responses . The pathogen structures that are recognized by TLR9 in bacterial or viral DNA are deoxycytidyl-deoxyguanosine dinucleotides (CpGs) in specific sequence contexts (CpG motifs) . The stimulatory activity of pathogen DNA can be mimicked by synthetic oligodeoxynucleotides (ODNs) containing such motifs (CpG ODNs) . The TLR9-mediated stimulation of the vertebrate innate immune system, and subsequently of the adaptive immune system, allows the use of TLR9 agonists as highly effective vaccine adjuvants for infectious diseases, and as stand-alone therapies or in combination with other therapies in cancer.

BMC Bioinformatics . 2004 Dec 15;5(1):198.
GenomeViz: visualizing microbial genomes; Ghai R et al.; BACKGROUND: An increasing number of microbial genomes are being sequenced and deposited in public databases . In addition, several closely related strains are also being sequenced in order to understand the genetic basis of diversity and mechanisms that lead to the acquisition of new genetic traits . These exercises have necessitated the requirement for visualizing microbial genomes and performing genome comparisons on a finer scale . We have developed GenomeViz to enable rapid visualization and subsequent comparisons of several microbial genomes in an interactive environment . RESULTS: Here we describe a program that allows visualization of both qualitative and quantitative information from complete and partially sequenced microbial genomes . Using GenomeViz, data deriving from studies on genomic islands, gene/protein classifications, GC content, GC skew, whole genome alignments, microarrays and proteomics may be plotted . Several genomes can be visualized interactively at the same time from a comparative genomic perspective and publication quality circular genome plots can be created . CONCLUSIONS: GenomeViz should allow researchers to perform visualization and comparative analysis of up to eight different microbial genomes simultaneously.

IEEE Trans Ultrason Ferroelectr Freq Control, 2004 Nov, 51(11), 1404 - 11
Pure shear horizontal SAW biosensor on langasite; Berkenpas E et al.; The undetected introduction of pathogens into food or water supplies can produce grave consequences in terms of economic loss and human suffering . Sensitive and selective sensors capable of quickly detecting microbial pathogens are urgently needed to limit the effects of bioterrorist incidents, accidents, or pollution . Shear horizontal surface acoustic wave (SH SAW) devices provide an attractive platform for the design of microbial biosensors that function in liquid media, where Rayleigh-type modes are rapidly attenuated . This paper reports on an exploratory SH SAW delay line designed and fabricated on langasite, La3Ga5SiO14 (LGS), along the novel Euler propagation direction (0 degrees, 22 degrees, 90 degrees) . A liquid chamber was fabricated and attached to the top surface, and the device was submitted to liquid and biochemical tests . Moderate (6 dB) additional attenuation of the transmission coefficient, /S21/, was consistently observed when the SH SAW delay line was assembled in the test fixture and submitted to the liquid tests, indicating that LGS is an attractive candidate for liquid sensing . Sensor selectivity can be achieved by integrating the LGS SH SAW delay line with a biochemical recognition layer . A test setup was implemented for the characterization of LGS SH SAW-based biosensors . The delay line response to biomolecule binding was shown by detection of sequential binding of proteins to the SH SAW device delay path . The biotinylated sensor was exposed sequentially to biotin-binding deglycosylated avidin, biotin-modified rabbit IgG, and goat anti-rabbit IgG antibody . As each protein was bound to the sensing surface, marked changes in the delay-line phase were recorded . The reported results demonstrate the capability of these devices to act as biochemical detectors in aqueous solutions, and this work represents the first effort using the novel material LGS in SAW-based biosensor technology.

Clin Exp Med, 2004 Dec, 4(3), 103 - 31
Behcet's disease; Al-Mutawa SA et al.; Behcet's disease is a recurrent multisystem vasculitis that can affect any organ or system, but was originally described as orogenital ulcerations and uveitis . Specific criteria have been proposed for diagnosis and identification of affected organs by different national authorities . Behcet's disease is proposed to be due to an antigen/antibody reaction . The antigen can be external (microbial or other) or self-antigen . Self-antigens include HLA-B, S and interphotoreceptor retinoid binding protein, the oral mucosal antigens, and alphatropomycin . The antibody reaction manifests as changes common to any inflammatory process . Circulating immune complexes and neutrophil hyperactivity are present with the expression of adhesion molecules related to disease activity associated with phenotypic and functional aberrations affecting both Th1 and Th2 activity . The results in an increase of interleukin-4, -10, and -1B and interleukin-2 receptors with deficient interleukin-12, interferon-gamma, and expression of cell adhesion CDII/CD18, accompanied by increased B cells secreting IgG . The clinical picture varies according to the organ affected; emphasis is placed on those of diagnostic importance and those affecting morbidity and mortality . The histopathology demonstrates vasculitis with inflammatory cell infiltration, the endothelium expressing ICAM-1 and E-selectin, vascular occlusion with sequences . Treatment includes corticosteroids . Other immunosuppressant and biological agents include anti-tumor necrosis factor, interferon-alpha, Campath-1, and the S antigen . Colchicine and other miscellaneous therapies are included.

J Clin Invest, 2004 Dec, 114(12), 1800 - 11
Utilizing the adjuvant properties of CD1d-dependent NK T cells in T cell-mediated immunotherapy; Silk JD et al.; Activation of invariant CD1d-dependent NK T cells (iNKT cells) in vivo through administration of the glycolipid ligand alpha-galactosylceramide (alpha-GalCer) or the sphingosine-truncated alpha-GalCer analog OCH leads to CD40 signaling as well as the release of soluble molecules including type 1 and gamma interferons that contribute to DC maturation . This process enhances T cell immunity to antigens presented by the DC . The adjuvant activity is further amplified if APCs are stimulated through Toll-like receptor 4, suggesting that iNKT cell signals can amplify maturation induced by microbial stimuli . The adjuvant activity of alpha-GalCer enhances both priming and boosting of CD8(+) T cells to coadministered peptide or protein antigens, including a peptide encoding the clinically relevant, HLA-A2-restricted epitope of the human tumor antigen NY-ESO-1 . Importantly, alpha-GalCer was used to induce CD8(+) T cells to antigens delivered orally, despite the fact that this route of administration is normally associated with blunted responses . Only T cell responses induced in the presence of iNKT cell stimulation, whether by the i.v . or oral route, were capable of eradicating established tumors . Together these data highlight the therapeutic potential of iNKT cell ligands in vaccination strategies, particularly "heterologous prime-boost" strategies against tumors, and provide evidence that iNKT cell stimulation may be exploited in the development of oral vaccines.

Bioinformatics . 2004 Dec 14; {Epub ahead of print}
The WHy domain mediates the response to desiccation in plants and bacteria; Ciccarelli FD et al.; MOTIVATION: The Hypersensitive Response (HR) is a process activated by plants after microbial infection . Its main phenotypic effects are both a programmed death of the plant cells nearby the infection site and a reduction of the microbial proliferation . Although many resistance genes (R genes) associated to HR have been identified, very few is known about the molecular mechanisms activated after their expression . RESULTS: The analysis of the product of one of the R genes, the Hin1 protein, led to the identification of a novel domain, which we named WHy because it is detectable in proteins involved in water stress and hypersensitive response . The expression of this domain during both biotic infection and response to desiccation points to a molecular machinery common to these two stress conditions . Moreover, its presence in a restricted number of bacteria suggests a possible use for marking plant pathogenicity . SUPPLEMENTARY INFORMATION: Supplementary data (Figure S1, Figure S2 and Table S1) and the alignment in clustal format are available at http://www.bork.embl.de/~ciccarel/WHy_add_data.html.

J Inorg Biochem, 2005 Jan, 99(1), 247 - 266
Nitric oxide dioxygenase function and mechanism of flavohemoglobin, hemoglobin, myoglobin and their associated reductases; Gardner PR; Microbial flavohemoglobins (flavoHbs) and hemoglobins (Hbs) show large ()NO dioxygenation rate constants ranging from 745 to 2900 muM(-1)s(-1) suggesting a primal ()NO dioxygenase (NOD) (EC 1.14.12.17) function for the ancient Hb superfamily . Indeed, modern O(2)-transporting and storing mammalian red blood cell Hb and related muscle myoglobin (Mb) show vestigial ()NO dioxygenation activity with rate constants of 34-89 muM(-1)s(-1) . In support of a NOD function, microbial flavoHbs and Hbs catalyze O(2)-dependent cellular ()NO metabolism, protect cells from ()NO poisoning, and are induced by ()NO exposures . Red blood cell Hb, myocyte Mb, and flavoHb-like activities metabolize ()NO in the vascular lumen, muscle, and other mammalian cells, respectively, decreasing ()NO signalling and toxicity . HbFe(III)-OO(), HbFe(III)-OONO and protein-caged {HbFe(III)-O()()NO(2)} are proposed intermediates in a reaction mechanism that combines both O-atoms of O(2) with ()NO to form nitrate and HbFe(III) . A conserved Hb heme pocket structure facilitates the dioxygenation reaction and efficient turnover is achieved through the univalent reduction of HbFe(III) by associated reductases . High affinity flavoHb and Hb heme ligands, and other inhibitors, may find application as antibiotics and antitumor agents that enhance the toxicity of immune cell-derived ()NO or as vasorelaxants that increase ()NO signalling.

Mol Plant Microbe Interact, 2004 Dec, 17(12), 1385 - 93
Fungal elicitation of signal transduction-related plant genes precedes mycorrhiza establishment and requires the dmi3 gene in Medicago truncatula; Weidmann S et al.; Suppressive subtractive hybridization and expressed sequence tag sequencing identified 29 plant genes which are upregulated during the appressorium stage of mycorrhiza establishment between Medicago truncatula J5 (Myc+) and Glomus mosseae . Eleven genes coding plant proteins with predicted functions in signal transduction, transcription, and translation were investigated in more detail for their relation to early events of symbiotic interactions . Expression profiling showed that the genes are activated not only from the appressorium stage up to the fully established symbiosis in the Myc+ genotype of M . truncatula, but also when the symbionts are not in direct cell contact, suggesting that diffusible fungal molecules (Myc factors) play a, role in the induction of a signal-transduction pathway . Transcript accumulation in roots of a mycorrhiza-defective Myc- dmi3 mutant of M . truncatula is not modified by appressorium formation or diffusible fungal molecules, indicating that the signal transduction pathway is required for a successful G . mosseae-M . truncatula interaction leading to symbiosis development . The symbiotic nodulating bacterium Sinorhizobium meliloti does not activate the 11 genes, which supposes early discrimination by plant roots between the microbial symbionts.

Clin Immunol, 2005 Jan, 114(1), 17 - 26
B-lymphocytes, innate immunity, and autoimmunity; Viau M et al.; Having evolved to generate a huge Ag-specific repertoire and to mount T cell-dependent responses and long-term memory, the B lymphocyte is a central player in the adaptive branch of immune defense . However, accumulating evidence indicates that B-1 cells of the peritoneal cavity and marginal zone (MZ) B cells of the spleen also can play innate-like immune functions . Their anatomical locations allow frequent Ag encounter . Secreting essentially germline-encoded, polyreactive Abs, and responding rapidly and vigorously to stimulation, these two B cell subsets have evolved to impart potentially protective responses . With their additional capacities to secrete factors that can directly mediate microbial destruction and to express Toll-like receptors (TLR), B cells provide an important link between the innate and adaptive branches of the immune system . Currently, the relevance of these innate-like B cells to the pathogenesis of autoimmune disease is the focus of investigation . In experimental models of autoimmunity, the sequestration of autoreactive B cells in the MZ has been proposed to be essential for the maintenance of self-tolerance . The low activation threshold of MZ B cells makes them particularly reactive to high loads and/or altered self-Ags, potentially exacerbating autoimmune disease . Their expansion in autoimmune models and their association with autoantibody secretion indicate that they may participate in tissue damage . The demonstration that B cell depletion therapies may represent a highly beneficial therapeutic goal in autoimmune disorders suggests that specific elimination of B-1 and MZ B cells may represent a more efficient immunointervention strategy in systemic autoimmunity.

Microbes Infect, 2004 Dec, 6(15), 1368 - 73
Integration of Toll-like receptor and phagocytic signaling for tailored immunity; Underhill DM et al.; Toll-like receptors (TLRs) are recruited to phagosomes and sample the contents of the compartment for microbial products . In this review, we discuss the relationship between phagocytosis and TLRs from two additional perspectives: first, TLR signaling modulates phagocytosis; second, phagocytosis modulates the consequences of TLR activation.

Drug Dev Ind Pharm, 2004, 30(10), 1069 - 78
Investigation of product quality between extemporaneously compounded progesterone vaginal suppositories and an approved progesterone vaginal gel; Mahaguna V et al.; The purpose of this investigation was to compare quality parameters, including product appearance, content uniformity, pH, weight uniformity, microbial limit testing and preservative effectiveness testing on extemporaneously compounded progesterone vaginal suppositories obtained from 10 randomly chosen compounding pharmacies (90 suppositories each) across the United States, to the Food and Drug Administration (FDA) approved prescription progesterone gel product (Prochieve/Crinone) which is manufactured in a cGMP regulated facility . The content uniformity and pH were determined using qualified methods . The microbial limits testing and preservative effectiveness testing were conducted according to compendial methods . Only one pharmacy provided suppositories that were all within the potency limits required for the prescription progesterone gel product . The other pharmacies provided at least some suppositories where progesterone content was either subpotent or superpotent for progesterone . The pH of most of the compounded suppository products was in the range of 4.22 to 7.68 with a median of 6.30 (normal vaginal pH is <5), whereas the gel product was 2.80 . For compounded product from one of the pharmacies, microbial limits testing indicated CDC group IVC-2 and Comamonas acidovorans were detected . This data indicates that pharmacy compounded delivery systems for progesterone should be used with caution.

Bioinformatics, 2003 Sep 1, 19(13), 1718 - 9
FindGDPs: identification of primers for labeling microbial transcriptomes for DNA microarray analysis; Blick RJ et al.; FindGDPs is a program that uses a greedy algorithm to quickly identify a set of genome-directed primers that specifically anneal to all of the open reading frames ina genome and that do not exhibit full-length complementarity to the members of another user-supplied set of nucleotide sequences.

J Zhejiang Univ Sci, 2005 Jan, 6(1), 1 - 3
On dendritic cell-based therapy for cancers; Onji M et al.; Dendritic cells (DCs), the most prevalent antigen-presenting cell in vivo, had been widely characterized in the last three decades . DCs are present in almost all tissues of the body and play cardinal roles in recognition of microbial agents, autoantigens, allergens and alloantigen . DCs process the microbial agents or their antigens and migrate to lymphoid tissues to present the antigenic peptide to lymphocytes . This leads to activation of antigen-specific lymphocytes . Initially, it was assumed that DCs are principally involved in the induction and maintenance of adaptive immune responses, but now it is evident that DCs also have important roles in innate immunity . These features make DCs very good candidates for therapy against various pathological conditions including malignancies . Initially, DC-based therapy was used in animal models of cancers . Data from these studies inspired considerable optimism and DC-based therapies was started in human cancers 8 years ago . In general, DC-based therapy has been found to be safe in patients with cancers, although few controlled trials have been conducted in this regard . Because, the fundamentals principles of human cancers and animal models of cancers are different, the therapeutic efficacy of the ongoing regime of DC-based therapy in cancer patients is not satisfactory . In this review, we covered the various aspects that should be considered for developing better regime of DC-based therapy for human cancers.

J Infect Dis, 2005 Jan 1, 191(1), 127 - 37 Epub 2004 Dec 02.
Glucuronoxylomannan, a microbial compound, regulates expression of costimulatory molecules and production of cytokines in macrophages; Monari C et al.; Glucuronoxylomannan (GXM) is a microbial compound that can modulate the immune response . We investigated (1) the receptors involved in uptake of GXM on monocyte-derived macrophages (MDMs) from healthy donors, (2) the effects of GXM on expression of specific receptors, (3) the effects of GXM mediated by pattern-recognition receptors, and (4) GXM modulation of MDM accessory and secretory functions . Cellular receptors involved in uptake of GXM included Fc gamma RII, CD18, Toll-like receptor (TLR) 4, and CD14 . Some biological functions of MDMs were profoundly affected by treatment with GXM, resulting in (1) increased expression of CD40 and CD86 via perturbation of TLR4, (2) decreased expression of major histocompatibility complex class II, (3) induction of interleukin-10 but not of tumor necrosis factor-alpha, and (4) decreased lipopolysaccharide (LPS)-induced production of cytokines . GXM represents an attractive compound to limit inflammatory processes and induce an LPS-tolerant state.

J Mol Med, 2005 Jan, 83(1), 12 - 25 Epub 2004 Dec 11.
Autoantigen complementarity: a new theory implicating complementary proteins as initiators of autoimmune disease; Pendergraft WF 3rd et al.; Autoimmune diseases affect approximately 1 in 21 persons in the United States . Treatment often requires long-term cytotoxic therapy . How and why these deleterious diseases occur is unclear . A serendipitous finding in our laboratory using serum from patients with autoimmune vasculitis led us to develop the theory of autoantigen complementarity, a novel concept that may elucidate the etiological and pathogenetic mechanisms underlying autoimmune disease in general . The theory proposes that the inciting immunogen that elicits a cascade of immunological events is not the self-antigen (the autoantigen) or its mimic but rather a protein that is complementary in surface structure to the autoantigen; that is, a protein homologous or identical to the amino acid sequence of translated antisense RNA from the noncoding strand of the autoantigen gene . The cascade begins when this complementary protein initiates the production of antibodies that in turn elicit an anti-antibody or anti-idiotypic response . These anti-idiotypic antibodies can now react with the autoantigen . Strikingly, homology search of complementary proteins yields microbial and fungal proteins, thus indicating that invading micro-organisms can deliver the inciting immunogen . Curiously, approximately 50% of our patients transcribe the complementary protein's antisense RNA . If it transpires that these aberrant RNAs are translated, the complementary protein would be produced by the individual . Here we review published research investigating complementary proteins, anti-idiotypic immune responses, and antisense transcripts, all of which support complementary proteins as initiators of autoimmune disease . In addition, we provide possible microbial and/or fungal organisms that may incite some of the most studied autoimmune diseases . Lastly, we propose mechanisms by which cell-mediated autoimmunity can be triggered by autoantigen complementarity . Based on our data and the contributions of the researchers described in this review, identification of proteins complementary to autoantigens is likely to be informative in most autoimmune diseases . This vein of study is in the early phases; however, we expect "autoantigen complementarity" is an underlying mechanism in many autoimmune diseases.

Springer Semin Immunopathol, 2005 Jan, 26(3), 221 - 9 Epub 2004 Nov 13.
Plasmacytoid dendritic cell precursors/type I interferon-producing cells sense viral infection by Toll-like receptor (TLR) 7 and TLR9; Ito T et al.; Plasmacytoid dendritic cell (pDC) precursors, also called type I IFN (alpha/beta/omega)-producing cells (IPCs), are the key effectors in the innate immune system because of their extraordinary capacity to produce type I IFNs against microbial infection, particularly viral infection . In contrast to myeloid DCs, human pDC/IPCs selectively express Toll-like receptor (TLR) 7 and TLR9 within the endosomal compartment . These receptors are specifically designed to recognize the nucleoside-based products derived from RNA viruses and DNA viruses . Therefore, this expression profile potentially enables pDC/IPCs to sense a variety of viruses . Stimulation of TLR7 or TLR9 leads to type I IFN responses through the MyD88 pathway . Thus, pDC/IPCs may play a central role in host defense against viral infection through the TLR7 and TLR9 system.

Transplantation, 2004 Dec 15, 78(11), 1597 - 600
Effect of mediators of innate immunity and inflammation on CD8+ veto cells; Zimring JC et al.; BACKGROUND: Methods of allotolerance induction that are successful in mice often fail in primates . Activators of innate immunity derived from microbial pathogens, which are present in primate populations but not in pathogen-free mouse colonies, have been shown to overcome tolerance induction by co-stimulatory blockade or regulatory T cells . Unlike other strategies, veto cells promote long-term allograft survival in primates . Accordingly, the authors hypothesized that veto cells are resistant to the effects of innate immune activation . METHODS: Using mixed lymphocyte reactions as a model of alloimmunization, the authors assessed the effect of inflammatory mediators on veto cell activity . RESULTS.: The authors demonstrate that the activity of veto cells is unaltered by lipopolysaccharide, double-stranded RNA, or interferon-gamma . Tumor necrosis factor-alpha modestly inhibited the veto effect, but only when veto cells were limiting . CONCLUSIONS: These findings may help to explain why veto cell-based therapies are more effective than other approaches in primate populations.

Hong Kong Med J, 2004 Dec, 10(6), 401 - 5
Microbial contamination of femoral head allografts; Chiu CK et al.; OBJECTIVE: To study the incidence of microbial contamination at the bone bank of the United Christian Hospital . DESIGN . Retrospective study . SETTING: Regional hospital, Hong Kong . PATIENTS: A total of 151 patients (33 men and 118 women) who underwent hip arthroplasty surgery and from whom femoral head allografts were retrieved between January 1994 and March 2000; and 81 patients in whom allografts were implanted . MAIN OUTCOME MEASURES: Bone biopsies were taken from the femoral head and used to detect any microbial contamination that might have occurred during removal and after storage . The rates of infection among recipients and donors were also assessed . RESULTS: Of the 151 allografts, 94 non-contaminated allografts were implanted by the end of the study . Fourteen (9.3%) heads showed positive culture results after retrieval and were discarded . Four (4.3%) of the 94 stored allografts that were implanted tested positive for microbial growth, but the recipients of these allografts did not develop any clinical infection . Three (3.2%) had wound infections after implantation of the stored allografts although the grafts had previously been tested negative for any microbial contamination . CONCLUSION: Our centre has a low allograft contamination rate . The wound infection rate among recipients was also low . The culture of a bone biopsy sample is a reliable method to detect contamination of bone grafts . However, the contamination rate among stored allografts should prompt orthopaedics departments to review allograft handling procedures, so as to minimise the chance of contamination.

Biol Reprod . 2004 Dec 8; {Epub ahead of print}
Association of Eppin with Semenogelin on Human Spermatozoa; Wang Z et al.; Eppin (SPINLW1; GeneID: 57119) is a single copy gene encoding a cysteine-rich protein found only in the testis and epididymis, which contains both Kunitz-type and WAP-type four disulfide core protease inhibitor consensus sequences . This study demonstrates that in seminal plasma and on human spermatozoa following ejaculation Eppin is bound to semenogelin I (Sg) . Six different experimental approaches: 1) Immunoprecipitation from spermatozoa and seminal plasma with anti-Eppin, 2) Co-localization in semen and spermatozoa, 3) Incubation of recombinant Eppin (rEppin) and rSg and immunoprecipitation with either anti-Eppin or anti-Sg, 4) Far-Western blotting of Eppin and Sg, 5) Saturation binding of (125)I-Sg to Eppin, which is competed by unlabeled Sg, and 6) direct binding of (125)I-Sg to Eppin on a blot, all demonstrate that Eppin and Sg bind to each other . To study the specificity of binding, recombinant fragments of Eppin and Sg were made and demonstrate that the Eppin(75-133) C-terminal fragment binds the Sg(164-283) fragment containing the only cysteine in human Sg I (Cys-239) . Reduction and carboxymethylation of Cys(239) blocks binding of (125)I-rEppin indicating that a disulfide bond may be necessary for Eppin binding . The physiological significance of the Eppin-semenogelin complex bound on the surface of ejaculate spermatozoa lies in its ability to provide anti-microbial activity for spermatozoa, which has been reported for both Eppin and semenogelin derived peptides, and in its ability to provide for the survival and preparation of spermatozoa for fertility in the female reproductive tract.

Microbiol Mol Biol Rev, 2004 Dec, 68(4), 686 - 91
Status of the microbial census; Schloss PD et al.; Over the past 20 years, more than 78,000 16S rRNA gene sequences have been deposited in GenBank and the Ribosomal Database Project, making the 16S rRNA gene the most widely studied gene for reconstructing bacterial phylogeny . While there is a general appreciation that these sequences are largely unique and derived from diverse species of bacteria, there has not been a quantitative attempt to describe the extent of sequencing efforts to date . We constructed rarefaction curves for each bacterial phylum and for the entire bacterial domain to assess the current state of sampling and the relative taxonomic richness of each phylum . This analysis quantifies the general sense among microbiologists that we are a long way from a complete census of the bacteria on Earth . Moreover, the analysis indicates that current sampling strategies might not be the most effective ones to describe novel diversity because there remain numerous phyla that are globally distributed yet poorly sampled . Based on the current level of sampling, it is not possible to estimate the total number of bacterial species on Earth, but the minimum species richness is 35,498 . Considering previous global species richness estimates of 10(7) to 10(9), we are certain that this estimate will increase with additional sequencing efforts . The data support previous calls for extensive surveys of multiple chemically disparate environments and of specific phylogenetic groups to advance the census most rapidly.

J Matern Fetal Neonatal Med, 2004 Sep, 16(3), 146 - 57
Fetal cardiac dysfunction in preterm premature rupture of membranes; Romero R et al.; Background: Preterm premature rupture of membranes (PROM) is associated with one-third of preterm births . In about 50% of preterm PROM cases, the fetuses will elicit a fetal inflammatory response syndrome (FIRS) . FIRS is associated with the impending onset of preterm labor, periventricular leukomalacia, neonatal sepsis, and long-term handicap, including the development of bronchopulmonary dysplasia and cerebral palsy . The fetal myocardium is a potential target organ of proinflam-matory cytokines released during FIRS . The objective of this study was to determine whether preterm PROM is associated with functional changes in the fetal heart, as determined by fetal echocardiography . Methods: A retrospective study was conducted to assess the diastolic function of fetuses with preterm PROM with documented microbial invasion of the amniotic cavity (n = 25), preterm PROM without microbial invasion of the amniotic cavity (n = 42), and fetuses from normal pregnancies (control group = 150) . Pregnancies with multiple gestation, fetal distress, fetuses that were small for gestational age, and major congenital anomalies were excluded . Fetal echocardiography studies were performed with two-dimensional ultrasound, color Doppler imaging and pulsed Doppler ultrasound . Non-parametric statistics were used for comparisons . A p value of < 0.05 was considered significant . Results: The prevalence of positive amniotic fluid cultures for micro-organisms in patients with preterm PROM was 35.8% (24/67) . Ureaplasma urealyticum was the most frequent isolate, either alone (41.7%; 10/24) or with other micro-organisms (29.2%; 7/24) . Fetuses with preterm PROM had a higher delta early diastolic filling/atrial contraction (E/A) peak velocity ratio, a higher delta E/A velocity-time integral (VTI) ratio, a lower delta A peak velocity, a lower delta A VTI, and a lower A VTI/total VTI ratio in the mitral valve compared to those with uncomplicated pregnancies . The delta E/A peak velocity ratio was significantly higher and the delta A VTI significantly lower in fetuses with preterm PROM and microbial invasion of the amniotic cavity than in those with preterm PROM without microbial invasion of the amniotic cavity . Conclusions: Preterm PROM is associated with changes in fetal cardiac function consistent with increased left ventricular compliance . These observations were also noted in fetuses with microbial invasion of the amniotic cavity . Our findings suggest that fetal cardiac function is altered in preterm PROM and, in particular, in cases with intra-amniotic infection.

J Microbiol Methods, 2005 Feb, 60(2), 207 - 16
Development of a novel compact sonicator for cell disruption; Borthwick KA et al.; Ultrasound microbial cell disrupters operating at around 20 kHz are often physically large and, due to significant heating, can be unsuitable for small sample volumes where biochemical integrity of the extracted product is required . Development of a compact device based on a 63.5-mm diameter, 6.5-mm thick tubular transducer for rapid cell disruption in small-volume samples in a high-intensity acoustic cavitation field with minimal temperature rises is described here . Suspensions of Saccharomyces cerevisiae were exposed to cavitation for various times in the compact device and a 20-kHz probe sonicator . Cell disruption was assessed by protein release and by staining . Yeast cell disruption was greater in the novel 267-kHz sonicator than in the 20-kHz probe sonicator for the same exposure time . A 1-dimensional (1-D) transfer matrix model analysis for piezoelectric resonators was applied to an axial cross-section of the tubular sonicator to predict frequencies of mechanical resonance in the sample volume associated with maximum acoustic pressure . Admittance measurements identified frequencies of electrical resonance . Ultrasonic cavitation noise peaks were detected by a hydrophone at both the mechanical and electrical resonances . Cell breakage efficiency was twice as great in terms of protein released per dissipated watt at the mechanical resonance predicted by the model, compared to those at the electrical resonance frequencies . The results form a basis for rational design of an ultrasound cell disruption technique for small-volume samples.

Sci Total Environ, 2005 Jan 5, 336(1-3), 91 - 103
Fatty acid profiling of microbial community during aging of mucilaginous aggregates in the northern Adriatic; Blazina M et al.; Aggregates differing in size and phytoplankton community composition were sampled in winter/spring 2001 and summer 1997 and 2002 (during mucilaginous event) in the northern Adriatic Sea . The fatty acid profiles (FAME) were determined in aggregates, bacterial population was grown from each aggregate and each bacterial isolate from aggregate's plated cultures . All aggregates irrespective of the season, aggregate size or phytoplankton community composition contained isolates from three distinct groups, with fatty acid profiles corresponding to alpha-Proteobacteria, gamma-Proteobacteria and Cytophaga-Flavobacter (CF) complex but in different relative proportions . Fatty acid profiles of the bacterial population grown from each aggregate revealed either the domination of one group (gamma, alpha or CF) or contribution of more groups (gamma and CF; alpha and CF) . Specifically, as the aggregates age a shift in favor of bacteria belonging to CF-complex and corresponding decrease in bacteria similar to that of Proteobacteria occur . During the aggregate aging process and degradation of the mucous matrix, besides bacterial succession, the phytoplankton growth took place inside the aggregates . Such an aging pattern was confirmed through laboratory experiments with aggregates inhabited by the diatom Cylindrotheca closterium.

Sci Total Environ, 2005 Jan 5, 336(1-3), 13 - 24
Model and field studies of the degradation of cross-linked polyacrylamide gels used during the revegetation of slate waste; Holliman PJ et al.; Cross-linked polyacrylamide gels are increasingly being used in environmental restoration schemes and horticulture as a means of enhancing water supply to plants . However, the environmental impact of cross-linked polyacrylamide gel deployment in soil remains poorly understood . This study assessed the chemical, physical and biological properties of new and field-conditioned cross-linked polyacrylamide gels . Both monomeric acrylamide (11 mug l(-1)) and acrylic acid (285 mug l(-1)) were observed in new gel; however, the levels of monomers in field-conditioned gels (1-6 years old) were very low (acrylamide <1 mug l(-1); acrylic acid <7 mug l(-1)) . Generally, freeze-thaw processes and exposure to UV radiation had little effect on gel acrylic acid and acrylamide concentrations . However, elevated temperatures (35 degrees C) caused a significant release of up to 144 mug l(-1) of acrylamide and 453 mug l(-1) of acrylic acid in new gel and up to 25 mug l(-1) of acrylamide and 157 mug l(-1) of acrylic acid in field-conditioned gels . In contrast, gel water holding capacity was highly dependent upon environmental conditions (UV exposure and freeze/thaw cycles produced the greatest loss of water holding in new gels) and gel age . Optical microscopy revealed that after placement in the field the gels became increasingly colonised over time by fungi and bacteria . In enrichment cultures, we were unable, however, to demonstrate microbial growth when cross-linked polyacrylamide was used as the sole nitrogen source . In summary, under a range of conditions cross-linked polyacrylamide did not release acrylamide above legally permitted limits, with the exception of gel subjected to elevated temperatures . However, their capacity for holding water decreased sharply within 18 months . We therefore conclude that cross-linked polyacrylamide placed in soil is relatively stable with respect to the production of potentially toxic acrylamide, a species with a short half-life, which degrades to the much less toxic acrylic acid . However, the loss of water holding capacity raises questions about its long-term effectiveness in land restoration schemes as this is the main reason it is used in this role.

J Endotoxin Res, 2004, 10(6), 413 - 8
Cell activation by Toll-like receptors: role of LBP and CD14; Finberg RW et al.; Members of the Toll-like receptor (TLR) family have been shown to be important in the activation of cells by a variety of microbial ligands . TLRs are thought to mediate the 'recognition event' that follows an encounter between a mammalian cell and a microbial agent . In the case of the response to bacterial lipopolysaccharide (LPS), it is clear that the ability of these cell surface proteins to initiate the events necessary for activation of cells to produce cytokines is dependent upon 'accessory proteins' such as the pattern recognition protein CD14 and the lipopolysaccharide binding protein (LBP) . While the role of these proteins in the LPS-specific response is defined, their role in other TLR responses has not been defined, but it is important in understanding these events and, potentially, in designing new therapeutic strategies . Here we report on the role of these proteins in the response to yeast zymosan . The requirements for this response (which unlike the response to LPS is a response to a particulate antigen) and the role of other serum proteins are defined.

J Endotoxin Res, 2004, 10(6), 406 - 12
Mechanisms of TLR9 activation; Latz E et al.; Non-methylated CpG-motifs in bacterial or viral DNA are recognized by TLR9 as foreign . The activation of TLR9 by microbial DNA or synthetic oligonucleotides based on these motifs leads to the induction of innate immune responses . We have compared the subcellular localization of fluorescent versions of TLR9 and TLR4 and found that TLR9 is expressed in the endoplasmic reticulum while TLR4 is expressed on the plasma membrane . Fluorescently tagged bacterial DNA or CpG-DNA was observed to traffic to a tubular lysosomal compartment in human pDCs . In stimulated cells, TLR9 translocated to CpG-DNA or microbial DNA containing structures in the endosome, where TLR9 binds to DNA and initiates signaling.

C R Biol, 2004 Sep-Oct, 327(9-10), 917 - 25
Oxalic acid: a microbial metabolite of interest for the pulping industry; Meyer-Pinson V et al.; Oxalate is a common metabolite produced by almost all plant-pathogenic fungi . The degradation of cell wall from poplar chips and poplar sawdust by oxalate is reviewed here . Oxalate treatments decrease slightly the amount of sugars constituting hemicelluloses, but only in fibres and not in sawdust or wood chips . The examination of the cell wall ultrastructure of wood chips by transmission electron microscopy (TEM) after polysaccharide staining showed a characteristic fading of the staining of the S1/S2 and S2/S3 transition areas, supporting the idea that reactivity and organization of polysaccharides had changed after the oxalate treatments . Finally, all these changes enhanced the ability of the wood chips to be defibrated by a thermomechanical (TMP) process, as well as the further refining of the pulps . Looking at the fiber surface, it became apparent that fracture areas during the TMP pulping had moved toward the S2 layer, explaining why defibrating and refining occurred more easily, with less energy input in the process.

Am J Rhinol, 2004 Sep-Oct, 18(5), 291 - 9
Nasal toxicity of benzalkonium chloride; Riechelmann H et al.; BACKGROUND: Benzalkonium chloride (BAC) is added to nasal preparations to prevent microbial contamination . Adverse effects of BAC on human nasal mucosa should be evaluated . METHODS: The ciliotoxicity of BAC was assessed in isolated human nasal epithelia from 15 donors . The effects of nasal BAC 0.05% (4 x 200 microL/day for 8 days) on nasal saccharin transport time, inflammatory cells and cytokine levels in nasal secretions, and nasal symptom scores were assessed in a randomized, double-blind crossover trial in 16 healthy volunteers . RESULTS: In vitro, BAC was ciliotoxic (p < 0.0001) . In vivo, BAC did not alter saccharin transport time in healthy individuals (p > 0.8) . No BAC-associated proinflammatory effects were observed . The staining index for myeloperoxidase was 4.8% in the placebo period and 6.3% (p = 0.42) in the BAC period . Also, nasal secretion levels of cytokines and the neuropeptide substance P revealed no BAC-associated differences . Concentrations for interleukin (IL)-6 in the placebo period were 41.5 pg/mL (0.9-91.7 pg/mL) and in the BAC period were 17.6 pg/mL (3.2-65.9 pg/mL; p = 0.46), and concentrations for substance P were 119 pg/mL (58-293 pg/mL) and 131 pg/mL (80-330 pg/mL; p = 0.31), respectively . Immediately after application, BAC caused nasal irritation (p = 0.001), a burning sensation (p = 0.0003), and hypersecretion (p = 0.006) . Moreover, BAC caused a persistent sensation of nasal irritation (p < 0.01) . CONCLUSION: BAC in concentrations used in nasal preparations is ciliotoxic . In healthy individuals, the ciliotoxic effect of BAC is neutralized, probably by components of nasal secretions . No BAC-related proinflammatory effects have been observed . At higher doses than normally used therapeutically, BAC caused significant nasal irritation.

J Immunol, 2004 Dec 15, 173(12), 7548 - 55
IL-10 released by concomitant TLR2 stimulation blocks the induction of a subset of Th1 cytokines that are specifically induced by TLR4 or TLR3 in human dendritic cells; Re F et al.; Recognition of microbial products through TLRs triggers the expression of several cytokines that regulate innate and adaptive immunity . Signaling by various TLRs is not equivalent and leads to differential gene induction . This study analyzed the responses of human dendritic cells (DCs) and PBMCs stimulated with agonists of TLR2, TLR3, TLR4, TLR5, and TLR7, first individually and then in combination . Several cytokines were equally induced by all TLR agonists, but four genes, IFN-beta, IFN-gamma-inducible protein 10 (IP-10), IL-12p35, and IL-15, showed a very restricted pattern of induction . Thus, each TLR appears to possess a distinctive ability to activate DCs or PBMCs, suggesting that TLR-mediated responses cannot be simply cataloged as resembling either TLR2 (MyD88 dependent) or TLR4 (MyD88 independent) and that other signaling modalities may exist . The analysis of DC and PBMC activation by combinations of TLR agonists revealed that TLR2 agonists are able to block the induction of IP-10, IL-12p35, and IFN-gamma, but not IL-15 and IFN-beta, by TLR3 and TLR4 . TLR2 stimulation led to rapid release of IL-10 that is responsible for inhibition of IP-10 and IL-12p35 induction . Cross-talk between different TLRs may modify the primary responses of TLR to their agonist, adding a further level of complexity to the regulation of innate immunity.

J Immunol, 2004 Dec 15, 173(12), 7471 - 80
Demonstration of the involvement of outer surface protein e coiled coil structural domains and higher order structural elements in the binding of infection-induced antibody and the complement-regulatory protein, factor H1; McDowell JV et al.; Factor H (fH) is an important regulator of the alternative complement cascade . Several human pathogens have been shown to bind fH to their surface, a process that facilitates immune evasion or cell to cell interaction . Among the pathogens that bind fH are some Borrelia species associated with Lyme disease and relapsing fever . The fH-binding proteins of the Lyme spirochetes form two classes (I and II) . In Borrelia burgdorferi B31MI, class I includes the outer surface protein E (OspE) paralogs, L39, N38, and P38, whereas the class II group includes A68 and additional proteins that have not yet been identified . To identify the OspE determinants involved in fH and OspE-targeting infection-induced Ab (iAb) binding, deletion, random, and site-directed mutagenesis of L39 were performed . Mutations in several different regions of L39 abolished fH and or iAb binding, indicating that separable domains and residues of OspE are required for ligand binding . Some of the mutants that lost the ability to bind fH, iAb, or both had only a single amino acid change . Site-directed mutagenesis of three putative coiled coil motifs of OspE revealed that these higher order structures are required for fH binding but not for iAb binding . The data presented within demonstrate that the binding of fH and iAb to the OspE protein is mediated by higher order structures and protein conformation . These studies advance our understanding of fH binding as a virulence mechanism and facilitate ongoing efforts to use fH-binding proteins in the development of microbial vaccines.

Int Immunol, 2005 Jan, 17(1), 1 - 14
Toll-like receptors in innate immunity; Takeda K et al.; Functional characterization of Toll-like receptors (TLRs) has established that innate immunity is a skillful system that detects invasion of microbial pathogens . Recognition of microbial components by TLRs initiates signal transduction pathways, which triggers expression of genes . These gene products control innate immune responses and further instruct development of antigen-specific acquired immunity . TLR signaling pathways are finely regulated by TIR domain-containing adaptors, such as MyD88, TIRAP/Mal, TRIF and TRAM . Differential utilization of these TIR domain-containing adaptors provides specificity of individual TLR-mediated signaling pathways . Several mechanisms have been elucidated that negatively control TLR signaling pathways, and thereby prevent overactivation of innate immunity leading to fatal immune disorders . The involvement of TLR-mediated pathways in autoimmune and inflammatory diseases has been proposed . Thus, TLR-mediated activation of innate immunity controls not only host defense against pathogens but also immune disorders.

J Anim Sci, 2005 Jan, 83(1), 187 - 95
Efficacy of using a combination of rendered protein products as an undegradable intake protein supplement for lactating, winter-calving, beef cows fed bromegrass hay; Encinias AM et al.; Seventy-two (36 in each of two consecutive years) lactating, British-crossbred cows (609 +/- 19 kg) were used to evaluate effects of feeding a feather meal-blood meal combination on performance by beef cows fed grass hay . Bromegrass hay (9.6% CP, DM basis) was offered ad libitum and intake was measured daily in individual Calan electronic headgates . Acclimation to Calan gates began approximately 20 d after parturition, and treatments were initiated 21 d later . Cows were assigned randomly to one of four treatments (DM basis) for 60 d: 1) nonsupplemented control (CON), 2) energy control (ENG; 790 g/d; 100% beet pulp), 3) degradable intake protein (DIP; 870 g/d; 22% beet pulp and 78% sunflower meal), or 4) undegradable intake protein (UIP; 800 g/d; 62.5% sunflower meal, 30% hydrolyzed feather meal, and 7.5% blood meal) . Net energy concentrations of supplements were formulated to provide similar NE(m) intakes (1.36 Mcal/d) . The DIP and UIP supplements were calculated to supply similar amounts of DIP (168 g/d) and to supply 64 and 224 g/d of UIP, respectively . Forage DMI (kg/d) decreased in supplemented vs . nonsupplemented (P = 0.03) and DIP vs . UIP (P = 0.001); however, when expressed as a percentage of BW, forage DMI was not different (P = 0.23) . Supplemented cows tended (P = 0.17) to lose less BW than CON . Body condition change was not affected (P = 0.60) by postpartum supplementation . No differences were noted in milk production (P = 0.29) or in calf gain during the supplementation period (P = 0.74) . Circulating insulin concentrations were not affected by treatment (P = 0.42) . In addition, supplementation did not affect circulating concentrations of NEFA (P = 0.18) or plasma urea nitrogen (P = 0.38) . Results of the current study indicate that supplementation had little effect on BW, BCS, milk production, or calf BW when a moderate-quality forage (9.6% CP) was fed to postpartum, winter-calving cows in optimal body condition (BCS > 5) . Supplemental UIP did not enhance cow performance during lactation . Forage UIP and microbial protein supply were adequate to meet the metabolizable protein requirements of lactating beef cows under the conditions of this study.

Int J Tuberc Lung Dis, 2004 Nov, 8(11), 1342 - 7
Comparison of two bacteriophage tests and nucleic acid amplification for the diagnosis of pulmonary tuberculosis in sub-Saharan Africa; Mbulo GM et al.; SETTING: National reference laboratory in Zambia, a high-incidence setting with a high prevalence of HIV infection . OBJECTIVE: To compare the performance of a commercial bacteriophage kit with a nucleic acid amplification kit and an 'in-house' bacteriophage method for rapid diagnosis of pulmonary tuberculosis (TB) . METHODS: Sputum specimens from suspected pulmonary TB cases were examined by direct fluorescence microscopy and culture on Lowenstein Jensen (LJ) . In a blinded study, remaining samples were tested by AMTD and FASTPlaqueTB or an in-house bacteriophage assay . Two specimen decontamination protocols were investigated . RESULTS: Microbial contamination of 40.4% was observed when using the FASTPlaqueTB kit specimen preparation protocol . When compared to culture on LJ, the sensitivity of the FASTPlaqueTB test was 20.7% . Implementation of a modified Petroff's decontamination protocol reduced contamination to 5.8% and the FASTPlaqueTB test detected 8/25 (32%) of culture-positive specimens . The sensitivity of AMTD and smear microscopy for these specimens were 64% and 48%, respectively . In a separate experiment the sensitivity of an in-house bacteriophage assay was 45.3% compared to 64.2% for AMTD and 45.3% for direct smear microscopy . CONCLUSIONS: Additional analysis of sputum specimens by bacteriophage assay provided no advantage in this setting . For the rapid diagnosis of TB, AMTD offered improved sensitivity over direct smear microscopy.

Water Sci Technol, 2004, 50(9), 33 - 40
Effect of the application of acid treated biosolids on marigold (Tagetes erecta) development; Diaz-Avelar J et al.; The use of biosolids for land restoration and crop production is a potential solution to improve food production worldwide . However, the microbial content usually restricts its application in crops that are consumed uncooked . An alternative practice is their use in floriculture . In this study, the effects of acid treated sludge on the development of marigold (Tagetes erecta) plants were evaluated under green house conditions . Biosolids were applied at the agronomic rate (AR) based on nitrogen requirements of the marigold . In addition, higher rates (10 and 20xAR) were applied to study their effect on the plants . Biosolids were mixed with tepetate (hard volcanic indurate layers) . Due to its origin, tepetate lacks nutrients and organic matter to adequately support plant development . The best treatment for marigold development was 10xAR, as plants reached an average height of 107 cm, with a growing speed of 1.01 cm/d, which is 20 times more than the control . Plants that received no biosolids produced 0.25 buds and 0.5 flowers per plant . In contrast, AR and 10xAR showed a production that ranged from 2 to 29 buds/plant and 4 to 15 flowers/plant, respectively . These results indicate the viability of reusing acid treated biosolids to improve marigold development.

Ophthalmic Surg Lasers Imaging, 2004 Nov-Dec, 35(6), 475 - 81
Cyclosporin-augmented laser peripheral iridoplasty; Crouch ER Jr et al.; BACKGROUND AND OBJECTIVE: Almost all patients develop iritis following argon laser peripheral iridoplasty . Numerous adverse effects, particularly elevated intraocular pressure (IOP) and reduced microbial resistance, complicate therapy with topical corticosteroids . An immunomodulator, such as cyclosporin A (CsA), avoids these undesirable effects, yet may suppress ocular inflammation . MATERIALS AND METHODS: Argon laser peripheral iridoplasty was performed on anesthetized rabbits with pigmented iris epithelium . Rabbits were randomly assigned to the untreated control, CsA (2%), or dexamethasone (0.1%) groups . Postoperative inflammation was documented by digital photography, IOP, and protein in aqueous humor . RESULTS: Iris injection, aqueous flare, and fibrin decreased most rapidly in the control group, as did protein in aqueous humor . Decreases in IOP of 49% to 58% were similar in all three groups . There were no differences in conjunctival congestion between the CONCLUSION: Neither treatment with antiinflammatory drugs that inhibit phagocytosis (e.g., topical steroids) nor treatment with anti-inflammatory drugs that suppress T-lymphocytes (e.g., topical sA) significantly attenuated inflammation following iridoplasty.

J Invertebr Pathol, 2004 Oct-Nov, 87(2-3), 94 - 104
Influence of temperature preferences of two Paecilomyces fumosoroseus lineages on their co-infection pattern; Fargues J et al.; In order to clarify the epidemiological potential of entomopathogenic fungi for insect pest control, the role of the temperature as one environmental constraint was investigated on the pattern of co-infection of Galleria mellonella by two distinct lineages of a hyphomycete, Paecilomyces fumosoroseus . The distribution of conidial populations collected on cadavers of hosts co-infected under 20 regimes, ranging from 13 to 35 degrees C, was examined . The apparent temperature tolerance of both fungal isolates was related to their in vitro colony growth and their in vivo sporulation ability . The conidial populations were characterized by molecular markers based on restriction fragment length polymorphisms of the internal transcribed spacers (ITS-RFLP) and random amplified polymorphic DNA (RAPD) contrasting profiles in combination with the conidial size . This study allowed a different temperature profile was identified for each isolate . Under most temperature regimes, only one lineage prevailed upon the infected insect; whereas both lineages coexisted at 20-25 and 25-25 degrees C . When one haplotype dominated, the displacement of the other one depended on its temperature tolerance . These results suggest that more consideration should be given to population-genetics analyses for evaluating the adaptability of microbial control agents to targeted environments.

Curr Pharm Des, 2004, 10(32), 4065 - 70
Drug resistance, virus fitness and HIV-1 mutagenesis; Chen R et al.; The evolution of antiretroviral drug resistance is a major problem in the treatment of human immunodeficiency virus type 1 (HIV-1) infection . Drug therapy failure is associated with accumulation of drug resistance mutations and results in the development of drug resistance . Drugs targeted against reverse transcriptase (RT) as well as drug-resistant RT have been shown to increase HIV-1 mutation frequencies . Furthermore, combinations of drug and drug-resistant RT can increase virus mutation frequencies in a multiplicative manner . The evolution of drug resistance also alters virus fitness . The correlation of increased HIV-1 mutation rates with the evolution of antiretroviral drug resistance indicates that drug failure could increase the likelihood of further resistance evolving from subsequent drug regimens . These observations parallel studies from microbial systems that provide evidence for a correlation between drug resistance development and increased pathogen mutation rates . Although increased mutant frequencies may be detrimental to effective therapy, the lethal mutagenesis of the HIV-1 genome may provide a new means for antiretroviral therapy.

Immunogenetics . 2004 Dec 1; {Epub ahead of print}
Identification and sequence analysis of chicken Toll-like receptors; Yilmaz A et al.; Toll-like receptors (TLRs) play an important role in the recognition of microbial components . Only chicken TLR2 and -4 have been reported in the literature . The objectives of this study were to identify new chicken TLRs and to evaluate evolutionary significance of these receptors . Searching chicken genomic databases and DNA sequencing revealed five new TLRs, TLR1 (type 1 and 2), -3, -5, and -7 . No chicken orthologues of mammalian TLR8, -9, or -10 were found . As in mammals, all chicken TLRs (chTLRs) share identical protein secondary structure that consists of several leucine-rich domains, a transmembrane domain, and Toll/Interleukin-1 receptor domain(s) . Phylogenetic analyses indicate that the identified chTLR genes are the orthologues of TLRs in mammals . Analyses of the number of synonymous substitutions per synonymous site and nonsynonymous substitutions per nonsynonymous site indicate that the nucleotide sequences coding for the leucine-rich repeats of chicken TLR1 type 1 and type 2 were significantly under positive Darwinian selection . In contrast, the sequences of other TLRs were under purifying selection . These results support the hypothesis that one of the major evolutionary strategies of the innate immune system is to recognize a few highly conserved microbial components with several conserved TLRs . The results also indicate that the sequence changes in the ligand-binding domains of TLR1 in chickens provide adaptive advantages during evolution.

Cell Death Differ, 2005 Feb, 12(2), 115 - 27
Externalization of host cell protein kinase C during enteropathogenic Escherichia coli infection; Crane JK et al.; Enteropathogenic Escherichia coli (EPEC) is a common cause of diarrhea in children in developing countries . Protein kinase C (PKC), a serine- and threonine-directed protein kinase, is rapidly activated following EPEC infection and this is accompanied by its translocation to a membrane-bound location where it is tightly bound to phosphatidylserine (PS) . EPEC infection causes host cell death, one of whose features is externalization of PS . We hypothesized that externalization of PS would be accompanied by externalization of PKC as well . We report that EPEC infection triggers the externalization of PKC to the outer surface of the host cell . Ecto-PKC remains firmly tethered to the cell but can be released by incubation with peptide or protein substrates for the enzyme . Ecto-PKC is intact and biologically active and able to phosphorylate protein substrates on the surface of the host cell . Phosphorylation of whole EPEC bacteria or EPEC-secreted proteins could not be detected . Externalization of PKC could be reproduced by the combination of an apoptotic stimulus (ultraviolet (UV) irradiation) and phorbol myristate acetate (PMA), a procedure which resulted in externalization of >25% of the total cellular content of PKC-alpha . In the presence of ATP, ecto-PKC inhibited UV-induced cell shrinkage, membrane blebbing, and propidium iodide uptake but not the activation of caspases 3 and 7 . This is the first report that expression of an ecto-protein kinase is altered by a microbial pathogen and the first to note that externalization of PKC can accompany apoptosis.Cell Death and Differentiation (2005) 12, 115-127 . doi:10.1038/sj.cdd.4401531 Published online 3 December 2004.

Nucleic Acids Res, 2004, 32(21), 6347 - 57 Print 2004.
Identification of genes with fast-evolving regions in microbial genomes; Zheng Y et al.; Complete sequences of multiple strains of the same microbial species provide an invaluable source for studying the evolutionary dynamics between orthologous genes over a relatively short time scale . Usually the intensity of the selection pressure is inferred from a comparison between the nonsynonymous substitution rate and the synonymous substitution rate . In this paper, we propose an alternative method for detecting genes with one or more fast-evolving regions from pairwise comparisons of orthologous genes . Our method looks for regions with overrepresented nonsynonymous mutations along the alignment, and requires a higher nonsynonymous evolution rate in those regions than the neutral evolution rate . It identifies gene targets under intensive selection pressure that are not detected from the conventional rate comparison analysis . For those identified genes with known annotations, most of them have a clear role in processes such as bacterial defense and host-pathogen interactions . Gene sets reported from our method provide a measure of the phenotypic divergence between two closely related genomes.

Am J Respir Cell Mol Biol . 2004 Dec 2; {Epub ahead of print}
Toll-like Receptor 4 or 2 Agonists Decrease Allergic Inflammation; Velasco G et al.; Toll-like receptors (TLRs) recognize highly conserved microbial molecular patterns, such as found in endotoxin . This study tested whether TLR4 and TLR2 stimulation in vivo would modulate subsequent adaptive (allergic) immune responses . We analyzed the effects of pulmonary administration of a TLR4 agonist, lipid A (LpA), and two TLR2 agonists, peptidoglycan (Ppg) and PamCys, in a murine model of allergic inflammation . The TLR agonists were administered during allergen sensitization or challenge . Both TLR agonists decreased the allergen-induced pulmonary recruitment of eosinophils when administered at sensitization or challenge . When given prior to sensitization, the TLR4 and TLR2 agonists decreased additional allergen-induced parameters of inflammation (pulmonary eosinophilia, bronchoalveolar lavage IL-13, total serum IgE, and airway hyperresponsiveness) . Interestingly, TLR4 and 2 agonists decreased the number of CD4(+) cells in the lung . Also, at the site of local allergen-stimulation, the draining thoracic lymph nodes, allergen-induced lymphocyte proliferation, and IL-13 secretion were decreased by administration of LpA and Ppg . These data provide a distinct example of the modulation of adaptive (allergic) responses by non-antigen-dependent stimuli . Our findings also demonstrate that both TLR4 and TLR2 agonists decrease allergic responses, supporting the concept that exposure to bacterial components under defined conditions may protect against allergic disease.

Biol Chem, 2004 Nov, 385(11), 997 - 1006
Molecular mechanism for activation and regulation of matrix metalloproteinases during bacterial infections and respiratory inflammation; Okamoto T et al.; Matrix metalloproteinases (MMPs) are critical mediators of tissue remodeling . Inappropriate regulation of MMPs causes many pathological events, including microbial invasion and inflammatory tissue damage . Some of the bacterial exoproteinases can effectively activate pro-MMPs (inactive zymogens) via limited proteolysis around their autoinhibitory domains . In addition, overproduction of nitric oxide (NO) may contribute to respiratory inflammation via the formation of reactive nitrogen species (RNS) . Several studies have identified regulatory properties of NO/RNS on biomolecules due to functional modification of their cysteine residues . In fact, NO/RNS can mediate activation and expression of MMPs, because RNS can interact with a cysteine switch in the autoinhibitory domain, thus converting proMMPs into their active forms without proteolysis . Many studies have indicated that NO/RNS can participate in expression of various genes that affect immune-inflammatory responses, including MMPs . Although NO in some cases upregulates MMPs, S -nitrosothiols downregulate MMP-9 expression by suppressing the NF-kappaB pathway . While microbial proteinases cause excessive activation of MMPs and contribute to microbial pathogenesis, NO/RNS may modulate expression and activation of MMPs as well as various inflammatory mediators, depending on the redox status at sites of inflammation . Therefore, appropriate regulation of MMPs may be of potential therapeutic value for various infections and inflammatory lung diseases.

Int J Hyg Environ Health, 2004 Oct, 207(5), 473 - 80
Water contamination in urban south India: household storage practices and their implications for water safety and enteric infections; Brick T et al.; Water contamination, at source and during household storage, is a major cause of enterically transmitted infections in developing countries . This study assessed contamination of the municipal water in a south Indian town, which obtains its water intermittently from a surface lake and by pumping subsurface water from a dry river bed, and monitored microbial contamination during household storage . All samples of the 'treated' municipal water were contaminated when freshly pumped, and on household storage, 25/37 (67%) showed increased contamination during storage periods from 1 to 9 days . Household storage in brass, but not in containers of other materials significantly decreased contamination of water (p = 0.04) . This was confirmed in the laboratory by testing water seeded with 10(3) to 10(5) Escherichia coli per 100 ml stored in containers of different materials (p < 0.01) . Despite the requirements for provision of safe drinking water in municipal areas, in practice the water supplied in Vellore is contaminated and current household storage practices increase the level of contamination in at least two-thirds of households . The implementation of locally appropriate point-of-use disinfection and safe household storage practices in developing countries is an urgent need to ensure a safe, reliable year-round supply in areas where clean water is not available.

Environ Sci Technol, 2004 Nov 1, 38(21), 5744 - 9
Fate of carbon tetrachloride during phytoremediation with poplar under controlled field conditions; Wang X et al.; The fate of carbon tetrachloride (CT) during phytoremediation with poplar was assessed by examining the transpiration of CT from leaves, diffusion from soil, tree trunks, and surface roots, and accumulation of chloride ion in soil and plant tissues . Feedwater containing 12-15 mg/L CT was added to the field test beds planted with poplar, and over 99% of the CT was removed . No significant amount of CT was transpired or diffused into the air, and no significant amount of CT-chlorine accumulated in the tree tissues . Chloride ion accumulated in the soil accounted for all of the CT-chlorine removed . When soils from the root zones were compared to unvegetated soils, microbial mineralization of CT was not enhanced in soils from the root zones as compared to unvegetated soils . Thus, we conclude that uptake and dechlorination of CT by plant tissues is likely the primary mechanism for phytoremediation by poplar.

Appl Environ Microbiol, 2004 Dec, 70(12), 7053 - 65
Sulfate-reducing bacteria in tubes constructed by the marine infaunal polychaete Diopatra cuprea; Matsui GY et al.; Marine infaunal burrows and tubes greatly enhance solute transport between sediments and the overlying water column and are sites of elevated microbial activity . Biotic and abiotic controls of the compositions and activities of burrow and tube microbial communities are poorly understood . The microbial communities in tubes of the marine infaunal polychaete Diopatria cuprea collected from two different sediment habitats were examined . The bacterial communities in the tubes from a sandy sediment differed from those in the tubes from a muddy sediment . The difference in community structure also extended to the sulfate-reducing bacterial (SRB) assemblage, although it was not as pronounced for this functional group of species . PCR-amplified 16S rRNA gene sequences recovered from Diopatra tube SRB by clonal library construction and screening were all related to the family Desulfobacteriaceae . This finding was supported by phospholipid fatty acid analysis and by hybridization of 16S rRNA probes specific for members of the genera Desulfosarcina, Desulfobacter, Desulfobacterium, Desulfobotulus, Desulfococcus, and Desulfovibrio and some members of the genera Desulfomonas, Desulfuromonas, and Desulfomicrobium with 16S rRNA gene sequences resolved by denaturing gradient gel electrophoresis . Two of six SRB clones from the clone library were not detected in tubes from the sandy sediment . The habitat in which the D . cuprea tubes were constructed had a strong influence on the tube bacterial community as a whole, as well as on the SRB assemblage.

Genome Res, 2004 Dec, 14(12), 2495 - 502
High-throughput mutation detection underlying adaptive evolution of Escherichia coli-K12; Honisch C et al.; Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis of base-specific cleavage products is an efficient, highly accurate tool for the detection of single base sequence variations . We describe the first application of this comparative sequencing strategy for automated high-throughput mutation detection in microbial genomes . The method was applied to identify DNA sequence changes that occurred in Escherichia coli K-12 MG1655 during laboratory adaptive evolution to new optimal growth phenotypes . Experiments were based on a genome-scale in silico model of E . coli metabolism and growth . This model computes several phenotypic functions and predicts optimal growth rates . To identify mutations underlying a 40-d adaptive laboratory evolution on glycerol, we resequenced 4.4% of the E . coli-K12 MG1655 genome in several clones picked at the end of the evolutionary process . The 1.54-Mb screen was completed in 13.5 h . This resequencing study is the largest reported by MALDI-TOF mass spectrometry to date . Ten mutations in 40 clones and three deviations from the reference sequence were detected . Mutations were predominantly found within the glycerol kinase gene . Functional characterization of the most prominent mutation shows its metabolic impact on the process of adaptive evolution . All sequence changes were independently confirmed by genotyping and Sanger-sequencing . We demonstrate that comparative sequencing by base-specific cleavage and MALDI-TOF mass spectrometry is an automated, fast, and highly accurate alternative to capillary sequencing.

Ying Yong Sheng Tai Xue Bao, 2004 Aug, 15(8), 1334 - 8
{Dynamics of microbial biomass C in a black soil under long-term fertilization and related affecting factors}; Li D et al.; The effect of long-term fertilization on the dynamics of microbial biomass C in a typical black soil of Northeastern China was studied in a field trail treated by different fertilizations . The results showed that the amount of soil microbial biomass C under different fertilizations varied significantly with growth stages . It was the highest in farmyard manure (M2 and M4) treatments, with a less seasonal fluctuation, second in NPK treatment, the peak at sowing period, and the lowest in CK, the peak at wax maturity stage . No significant correlation was found between the dynamic changes of soil microbial biomass C and soil biological, physical and chemical properties in all treatments, but the correlation of soil microbial biomass C with the contents of N, P and K in plants and that of crude protein in grain was significantly positive.

Nat Rev Immunol, 2004 Dec, 4(12), 978 - 88
The immunogenetics of asthma and eczema: a new focus on the epithelium; Cookson W; Asthma and eczema (atopic dermatitis) are the most common chronic diseases of childhood . These diseases are characterized by the production of high levels of immunoglobulin E in response to common allergens . Their development depends on both genetic and environmental factors . Over the past few years, several genes and genetic loci that are associated with increased susceptibility to asthma and atopic dermatitis have been described . Many of these genes are expressed in the mucosa and epidermis, indicating that events at epithelial-cell surfaces might be driving disease processes . This review describes the mechanisms of innate epithelial immunity and the role of microbial factors in providing protection from disease development . Understanding events at the epithelial-cell surface might provide new insights for the development of new treatments for inflammatory epithelial disease.

Nat Rev Immunol, 2004 Dec, 4(12), 953 - 64
War and peace at mucosal surfaces; Sansonetti PJ; That we live with numerous bacteria in our gut without any adverse effects is a remarkable feat by the body's immune system, particularly considering the wealth of sensing and effector systems that are available to trigger inflammatory or innate immune responses to microbial intrusion . So, a fine line seems to exist between the homeostatic balance maintained in the presence of commensal gut flora and the necessarily destructive response to bacterial pathogens that invade the gut mucosa . This review discusses the mechanisms for establishing and controlling the 'dialogue' between unresponsiveness and initiation of active immune defences in the gut . Si vis pacem, para bellum . (If you wish for peace, prepare for war.).

Environ Pollut, 2005 Mar, 134(1), 57 - 69
Do tubificid worms influence the fate of organic matter and pollutants in stormwater sediments?
Mermillod-Blondin F, Nogaro G, Datry T, Malard F, Gibert J.
In urban area, management of stormwater leads to the accumulation of polluted sediments at the water-sediment interface of various aquatic ecosystems . In many cases, these sediments are colonised by dense populations of tubificid worms . However, the influence of tubificid worms on the fate of stormwater sediments has never been tackled . The aim of this study was to measure in sediment columns the influence of tubificid worms on sediment reworking, organic matter processing (O(2) uptake and release of NH(4)(+), NO(3)(-), PO(4)(3-), and dissolved organic carbon), release of hydrocarbons and heavy metals, and microbial characteristics . Results showed that tubificid worms increased the release of NH(4)(+), PO(4)(3-), and dissolved organic carbon by 2-, 4-, and 3-fold, respectively . O(2) uptake also increased by more than 35% due to tubificid activity . The increase in the percentages of active bacteria and hydrolytic activity in the presence of worms indicated that the higher sediment respiration was caused by the stimulation of microbial communities . A reduction of the number of sulphate-reducing bacteria in the uppermost layers of the sediment was attributed to the penetration of O(2) due to worm activity . These significant effects of tubificid worms were probably linked to the dense network of burrows, which enhanced the exchange surface between the water column and the sediment . No release of heavy metals and hydrocarbons to the water phase was detected in the sediment columns . Understanding the fate and effect of organic stormwater sediments in the natural environment requires the integration of the role of bioturbation in urban pollution studies.

J Nutr, 2004 Dec, 134(12), 3378 - 84
Development of an assay to quantify rumen ciliate protozoal biomass in cows using real-time PCR; Sylvester JT et al.; Currently used microbial markers cannot distinguish protozoal nitrogen (N) from bacterial N, thus limiting research on protozoal quantification in vivo by the lack of a repeatable, accurate marker for protozoal N . We report the development of a real-time PCR assay targeting the gene encoding 18S rDNA to quantify the amount of protozoal biomass in ruminal fluid and duodenal digesta . Protozoal cells were harvested from rumen fluid and concentrated for evaluation of recovery of rDNA in samples from the rumen and the duodenum . The DNA from concentrated cells was extracted with virtually 100% efficiency both before and after column purification . After serial spiking of protozoal cells into duodenal fluid over the entire range of quantification, the recovery was highly linear and constant at 81% . After serially spiking increasing quantities of protozoal rDNA into a constant volume of duodenal samples, nonlinear regression verified constant recovery of background rDNA in duodenal samples regardless of the ratio of target:nontarget rDNA . Recommendations for the procedure, including replication per sample, are described herein.

Front Biosci, 2005 Jan 1, 10, 743 - 50 Print 2005 Jan 1.
HIV-1 mutagenesis during antiretroviral therapy: implications for successful drug treatment; Chen R et al.; The evolution of antiretroviral drug resistance is a major problem in the treatment of human immunodeficiency virus type 1 (HIV-1) infection . Drug therapy failure is associated with accumulation of mutations and results in the development of drug resistance . Drugs targeted against reverse transcriptase (RT) as well as drug-resistant RT have been shown to increase HIV-1 mutation frequencies . Furthermore, combinations of drug and drug-resistant RT can increase virus mutation frequencies in a multiplicative manner . The evolution of drug resistance also alters virus fitness . The correlation of increased HIV-1 mutation rates with the evolution of antiretroviral drug resistance indicates that drug failure could increase the likelihood of further resistance evolving from subsequent drug regimens . These observations parallel studies from microbial systems that provide evidence for a correlation between drug resistance development and increased pathogen mutation rates . Although increased mutant frequencies may be detrimental to effective therapy, the lethal mutagenesis of the HIV-1 genome may provide a new means for antiretroviral therapy.

Eksp Klin Gastroenterol, 2004, (4), 29 - 32, 108
{State of humoral immunity and phagocytic activity of neutrophils in patients with ulcer and influence of low-intensity laser therapy}; A generalized plate method for estimating total aerobic microbial count; ALZA Corporation, 1950 Charleston Road, Mountain View, CA 94043, USA . kho@alzus.jnj.com

The plate method outlined in Chapter 61: Microbial Limit Tests of the U.S . Pharmacopeia (USP 61) provides very specific guidance for assessing total aerobic bioburden in pharmaceutical articles . This methodology, while comprehensive, lacks the flexibility to be useful in all situations . By studying the plate method as a special case within a more general family of assays, the effects of each parameter in the guidance can be understood . Using a mathematical model to describe the plate counting procedure, a statistical framework for making more definitive statements about total aerobic bioburden is developed . Such a framework allows the laboratory scientist to adjust the USP 61 methods to satisfy specific practical constraints . In particular, it is shown that the plate method can be conducted, albeit with stricter acceptance criteria, using a test specimen quantity that is smaller than the 10 g or 10 mL prescribed in the guidance . Finally, the interpretation of results proffered by the guidance is re-examined within this statistical framework and shown to be overly aggressive.

PDA J Pharm Sci Technol, 2004 Sep-Oct, 58(5), 254 - 60
Environmental monitoring: data trending using a frequency model; Caputo RA et al.; Environmental monitoring programs for the oversight of classified environments have used traditional statistical control charts to monitor trends in microbial recovery for classified environments . These methodologies work well for environments that yield measurable microbial recoveries . However, today successful increased control of microbial content yields numerous instances where microbial recovery in a sample is generally zero . As a result, traditional control chart methods cannot be used appropriately . Two methods to monitor the performance of a classified environment where microbial recovery is zero are presented . Both methods use the frequency between non-zero microbial recovery as an event . Therefore, the frequency of events is monitored rather than the microbial recovery count . Both methods are shown to be appropriate for use in the described instances.

Ir Med J, 2004 Oct, 97(9), 278 - 80
The burden of severe pneumococcal infection in Ireland: potential effectiveness and indirect benefits of the 7-valent pneumococcal conjugate vaccine; McCormack S et al.; This study attempts to overcome the problem of under-reporting of serious pneumococcal infection in the Republic of Ireland by adding a proportion of 'unspecified' disease to pneumococcal disease reports . ICD-9 data for all age groups was collected on meningitis (from the National Disease Surveillance Centre), and septicaemia and pneumonia (from the Hospital In-Patient Enquiry system) for the year 1999 . A 7-valent pneumococcal conjugate vaccine (PCV) has been shown to have significant effectiveness in the target paediatric age group and also indirect herd effects on the US adult population . The implications of these direct and indirect effects were applied to the epidemiology of serious pneumococcal infection in Ireland . The annual reported incidence rates for laboratory confirmed severe pneumococcal disease in Ireland in 1999 may underestimate both the morbidity and mortality of disease by 21% and 28% respectively . In all age groups 1,183 cases and 132 deaths may be prevented annually by the introduction of the vaccine . In addition, the vaccine provides an effective new tool for reducing disease caused by antibiotic resistant pneumococci thus assisting in the control of anti-microbial resistance in humans.

Science, 2004 Nov 26, 306(5701), 1565 - 7
Periodical cicadas as resource pulses in North American forests; Yang LH; Resource pulses are occasional events of ephemeral resource superabundance that occur in many ecosystems . Aboveground consumers in diverse communities often respond strongly to resource pulses, but few studies have investigated the belowground consequences of resource pulses in natural ecosystems . This study shows that resource pulses of 17-year periodical cicadas (Magicicada spp.) directly increase microbial biomass and nitrogen availability in forest soils, with indirect effects on growth and reproduction in forest plants . These findings suggest that pulses of periodical cicadas create "bottom-up cascades," resulting in strong and reciprocal links between the aboveground and belowground components of a North American forest ecosystem.

Science, 2004 Nov 26, 306(5701), 1517 - 9
Natural killer cell signaling pathways; Vivier E et al.; Natural killer (NK) cells are lymphocytes of the innate immune system that are involved in the early defenses against foreign cells, as well as autologous cells undergoing various forms of stress, such as microbial infection or tumor transformation . NK cell activation is controlled by a dynamic balance between complementary and antagonistic pathways that are initiated upon interaction with potential target cells . NK cells express an array of activating cell surface receptors that can trigger cytolytic programs, as well as cytokine or chemokine secretion . Some of these activating cell surface receptors initiate protein tyrosine kinase (PTK)-dependent pathways through noncovalent associations with transmembrane signaling adaptors that harbor intracytoplasmic ITAMs (immunoreceptor tyrosine-based activation motifs) . Additional cell surface receptors that are not directly coupled to ITAMs also participate in NK cell activation . These include NKG2D, which is noncovalently associated to the DAP10 transmembrane signaling adaptor, as well as integrins and cytokine receptors . NK cells also express cell surface inhibitory receptors that antagonize activating pathways through protein tyrosine phosphatases (PTPs) . These inhibitory cell surface receptors are characterized by intracytoplasmic ITIMs (immunoreceptor tyrosine-based inhibition motifs) . The tyrosine-phosphorylation status of several signaling components that are substrates for both PTKs and PTPs is thus key to the propagation of the NK cell effector pathways . Understanding the integration of these multiple signals is central to the understanding and manipulation of NK cell effector signaling pathways.

Science, 2004 Nov 26, 306(5701), 1509 - 11
Jekyll and Hyde in the microbial world; Truckses DM et al.; Fungi are nonmotile organisms that obtain carbon from compounds in their immediate surroundings . Confronted with nutrient limitation, the yeast Saccharomyces cerevisiae undergoes a dimorphic transition, switching from spherical cells to filaments of adherent, elongated cells that can invade the substratum . A complex web of sensing mechanisms and cooperation among signaling networks (including a mitogen-activated protein kinase cascade, cyclic adenosine monophosphate-dependent protein kinase, and 5'-adenosine monophosphate-activated protein kinase) elicits the appropriate changes in physiology, cell cycle progression, cell polarity, and gene expression to achieve this differentiation . Highly related signaling processes control filamentation and virulence of many human fungal pathogens.

J Microbiol Methods, 2005 Jan, 60(1), 131 - 3
A simple and economic preservation method for genomic bacterial DNA from clinically significant pathogens; Moore JE et al.; Bacterial culture was allowed to dry to completeness on Columbia agar base with defibrinated horse blood . Following 6 months storage at room temperature, microbial DNA was extracted and successfully amplified by PCR . This storage technique has the advantage over other methods of not requiring (i) a DNA extraction protocol prior to storage and (ii) refrigeration and/or freezing . This technique maybe useful in the transportation of bacterial genomic DNA in nonviable cells as well as reliable method for the storage of DNA in underdeveloped countries.

J Microbiol Methods, 2005 Jan, 60(1), 83 - 92
Capillary electrophoresis single-strand conformation polymorphism analysis for monitoring soil bacteria; King S et al.; The ability to effectively monitor a microbial community is necessary to design and implement remediation strategies for contaminated soil . Single-strand conformation polymorphism (SSCP), a technique which separates DNA fragments based on their sequence, was used to analyze amplified 16S rRNA gene fragments of 12 common soil bacteria . Separation was performed using capillary electrophoresis (CE), as opposed to other common gel techniques, to eliminate the need for band analysis on gel matrices . Four different universal bacterial primer sets were used for DNA amplification: 341-534, P11-P13, Er10-Er11, and Er14-Er15 corresponding to the V3, V8, V2, and V4 regions, respectively . The forward strand of each primer was labeled with 6-carboxy fluorescein fluorescent dye . Analyses were performed on the Applied Biosystems 310 genetic analyzer using GeneScan Analysis Software version 3.5 . The best results were obtained using primer 341-534, in which 6 of the 12 bacteria could be distinguished . By combining primer sets 341-534 and Er10-Er11, all 12 of the bacteria could be separated, indicating various degrees of polymorphism within the selected primer regions . When performing simultaneous amplification and analysis of all 12 species some preferential amplification occurred, as not all peaks could be observed . However, SSCP profiles obtained for pure bacterial cultures show the potential of CE-SSCP for bacterial community analysis.

J Microbiol Methods, 2005 Jan, 60(1), 63 - 71
Culture independent PCR: an alternative enzyme discovery strategy; Jacobsen J et al.; Degenerate primers were designed for use in a culture-independent PCR screening of DNA from composite fungal communities, inhabiting residues of corn stovers and leaves . According to similarity searches and alignments amplified clone sequences affiliated with glycosyl hydrolase family 7 and glycosyl hydrolase family 45 though significant sequence divergence was observed . Glycosyl hydrolases from families 7 and 45 play a crucial role in biomass conversion to fuel ethanol . Research in this renewable energy source has two objectives: (i) To contribute to development of a renewable alternative to world's limited crude fossil oil reserves and (ii) to reduce air pollution . Amplification with 18S rDNA-specific primers revealed species within the ascomycetous orders Sordariales and Hypocreales as well as basidiomycetous order Agaricales to be present in these communities . Our study documents the value of culture-independent PCR in microbial diversity studies and could add to development of a new enzyme screening technology.

Clin Diagn Virol, 1996 Feb, 5(1), 21 - 6
Evaluation of 15 commercial enzyme immunoassays for the detection of rubella-specific IgM; Hudson P et al.; Background: Detection of rubella-specific IgM is a critical investigation in the diagnosis of recent or congenital rubella for which many commercial enzyme immunoassays (EIAs) are now available . Objectives: To evaluate 15 commercially-available EIA kits for the detection of rubella-specific IgM . Study design: A panel of 229 sera was established comprising 72 sera from cases of primary rubella, re-infection, congenital rubella and primary immunization in which rubella-specific IgM had been detected by M-antibody capture radioimmunoassay (MACRIA); 80 sera, negative by MACRIA and no evidence of recent illness; 77 sera from recent illness with a number of other microbial agents, sera containing rheumatoid factor, and sera previously shown to give non-specific reactivity by MACRIA . These sera were tested in 15 commercial EIA kits for rubella-specific IgM according to the manufacturers' instructions . Sensitivity and specificity for each assay was established . Results: Specificities ranged from 82-98%, improving to 88-100% if the previously established non-specific reactive sera were excluded . Sensitivities ranged from 63-92% . Although only two sera from congenital rubella were tested, for one serum only four assays detected specific IgM . Conclusion: Some commercial EIAs for rubella-specific IgM now give an acceptable balance of sensitivity and specificity, but all assays may give reactivity with occasional sera where there is no supporting evidence for recent primary rubella or rubella reinfection.

Infect Control Hosp Epidemiol, 2004 Nov, 25(11), 1000 - 1
Colonization of personal digital assistants carried by healthcare professionals; Hassoun A et al.; This study evaluated personal digital assistant (PDA) microbial colonization before and after cleaning with alcohol . Samples from 75 PDAs were processed . Before cleaning, 96% of the samples were culture positive . After cleaning, 75% became culture negative . PDAs cleaned with an alcohol swab demonstrated significant reduction in colonization.

FEMS Microbiol Lett, 2004 Nov 15, 240(2), 181 - 6
RNA fingerprinting of microbial community in the rhizosphere soil of grain legumes; Sharma S et al.; Microbial structural and expression profiles of the rhizospheres of three legumes, faba beans, peas and white lupin, were compared by RNA-arbitrarily primed PCR technique . Two different primers, M13 reverse and 10-mer primers, were used in the amplification and products resolved on non-denaturing polyacrylamide gel . With both DNA and RNA profiles Lupinus and Pisum rhizospheres were more similar to each other than to Vicia rhizosphere . The RAP-PCR products were also dot blotted and probed for bacterial peptidase transcripts . Plant-dependent rhizosphere effect was evident by the marked absence of transcripts for bacterial neutral metallopeptidase in Lupinus rhizosphere . The results of dot blot were further confirmed by RT-PCR for the expression of bacterial neutral metallopeptidase in the three rhizospheres.

Can J Gastroenterol, 2004 Nov, 18(11), 687 - 90
Granulomatous bronchiolitis with necrobiotic pulmonary nodules in Crohn's disease; Freeman HJ et al.; A 37-year-old man with extensive Crohn's disease of the stomach, small and large intestine for almost a decade developed respiratory symptoms and radiological findings suggestive of pneumonia that failed to resolve with antibiotic treatment . Computed tomography scanning of his lungs showed extensive changes with cavitated parenchymal nodules . Histological evaluation of an open lung biopsy showed granulomatous bronchiolitis and pulmonary necrobiosis . Treatment with steroids and immunosuppression resulted in complete resolution of his clinical symptoms of pneumonia and abnormal computed tomography imaging changes . Granulomatous bronchiolitis and necrobiotic nodules may be a manifestation of Crohn's disease in the absence of microbial agents, including mycobacteria or fungal agents . While a multiplicity of complex pulmonary changes may occur in Crohn's disease, their clinical recognition and precise pathological definition may be particularly important if treatment with a biological agent, such as infliximab, is being considered.

Ann Nutr Metab . 2004 Nov 22;48(6):369-373 {Epub ahead of print}
Indigestible Components of Grape Seeds Modify Cecal Enzyme Activity in Rats; Goni I; Background: The effect of grape seed indigestible fraction (GSIF) on cecal enzyme activity was studied . Methods: beta-Glucuronidase, beta-glucosidase, azoreductase, nitroreductase and nitrate reductase were measured in the cecal content of adult Wistar rats fed with a fiber-free diet supplemented with 5% cellulose or 5% GSIF as the source of dietary fiber for 4 weeks . Results: The intake of GSIF did not affect body weight gain nor food intake . However, GSIF caused a significant increase in cecal content, cecal wall and fresh and dry stool weight . Bacterial enzyme activities were lower in the GSIF-fed group than in the cellulose-fed group, although the difference was also significant for nitroreductase and beta-glucuronidase . Conclusion: Adaptation to a diet containing GSIF led to changes in microbial activity that involved a decrease in reductive and hydrolytic enzymatic activities implicated in the conversion of procarcinogens into carcinogens . Copyright (c) 2004 S . Karger AG, Basel.

Plant Cell Physiol, 2004 Oct, 45(10), 1347 - 60
New TAXI-type xylanase inhibitor genes are inducible by pathogens and wounding in hexaploid wheat; Igawa T et al.; TAXI-I (Triticum aestivum xylanase inhibitor I) is a wheat grain protein that inhibits arabinoxylan fragmentation by microbial endo-beta-1,4-xylanases used in the food industry . Although TAXI was speculated to be involved in counterattack against pathogens, there is actually no evidence to support this hypothesis . We have now demonstrated the presence of TAXI family members with isolation of two mRNA species, Taxi-III and Taxi-IV . At the nucleotide sequence level, Taxi-III and Taxi-IV were 91.7% and 92.0% identical, respectively, to Taxi-I, and Taxi-III and Taxi-IV were 96.8% identical . Accumulation of Taxi-III/IV transcripts was most evident in roots and older leaves where transcripts of Taxi-I were negligible . When challenged by fungal pathogens Fusarium graminearum and Erysiphe graminis, the concentrations of Taxi-III/IV transcripts increased significantly . In contrast, the increases in Taxi-I transcripts in response to these pathogens were rather limited . Both Taxi-I and Taxi-III/IV were strongly expressed in wounded leaves . The upstream region of Taxi-III contained W boxes and GCC boxes, which are sufficient to confer pathogen and wound inducibility on promoters . Recombinant TAXI-III protein inhibited Aspergillus niger and Trichoderma sp . xylanases: it was also active against some spelt xylan-induced xylanases of F . graminearum . These features suggest that some, but not all, TAXI-type xylanase inhibitors have a role in plant defense.

Drugs, 2004, 64(24), 2763 - 92
Viral prophylaxis in organ transplant patients; Slifkin M et al.; Viral pathogens have emerged as the most important microbial agents having deleterious effects on solid organ transplant (SOT) recipients . Antiviral chemoprophylaxis involves the administration of medications to abort transmission of, avoid reactivation of, or prevent progression to disease from, active viral infection . Cytomegalovirus (CMV) is the major microbial pathogen having a negative effect on SOT recipients . CMV causes infectious disease syndromes, augments iatrogenic immunosuppression and is commonly associated with opportunistic superinfection . CMV has also been implicated in the pathogenesis of rejection . Chemoprophylactic regimens for CMV have included oral aciclovir (acyclovir) at medium and high doses, intravenous and oral ganciclovir, and the prodrugs valaciclovir (valacyclovir) and valganciclovir . CMV prophylactic strategies should be stratified, with the highest-risk patients receiving the most 'potent' prophylactic regimens . Herpes simplex virus (HSV) reactivation in SOT recipients is more frequent, may become more invasive, takes longer to heal, and has greater potential for dissemination to visceral organs than it does in the immunocompetent host . Prophylactic regimens for CMV are also effective chemoprophylaxis against HSV; in the absence of CMV prophylaxis, aciclovir, valaciclovir or famciclovir should be used as HSV prophylaxis in seropositive recipients . Primary varicella-zoster virus (VZV) after SOT is rare and most commonly seen in the paediatric transplant population because of VZV epidemiology . Zoster occurs in 5-15% of patients, usually after the sixth post-transplant month . Prophylactic regimens for zoster are neither practical nor cost effective after SOT because of the late onset of disease and low proportion of affected individuals . All SOT recipients should receive VZV immune globulin after contact with either varicella or zoster . Epstein-Barr virus has its most significant effect in SOT as the precipitating factor in the development of post-transplant lymphoproliferative disorders . Antiviral agents that could be effective are the same as those used for CMV, but indications for and effectiveness of prophylaxis are poorly established . Hepatitis B virus (HBV) and hepatitis C virus (HCV) are important pathogens in the SOT population as indications for transplantation . So-called 'prophylaxis' for recurrent HBV and HCV after liver transplantation is controversial, suppressive rather than preventive, and potentially lifelong . Influenza infection after SOT is acquired by person-to-person contact . During epidemic periods of influenza, transplant populations experience a relatively high frequency of infection, and influenza may affect immunosuppressed SOT recipients more adversely than immunocompetent individuals . Antiviral medications for prevention of influenza are administered as post-exposure prophylaxis to SOT recipients, in addition to yearly vaccine, in circumstances such as influenza epidemics and nosocomial outbreaks, and after exposure to a symptomatic individual during 'flu season'.

J Parasitol, 2004 Oct, 90(5), 1150 - 4
Quantification of Leishmania infantum parasites in tissue biopsies by real-time polymerase chain reaction and polymerase chain reaction-enzyme-linked immunosorbent assay; Rolao N et al.; Most of the experimental studies of Leishmania spp . infection require the determination of the parasite load in different tissues . Quantification of parasites by microscopy is not very sensitive and is time consuming, whereas culture microtitrations remain laborious and can be jeopardized by microbial contamination . The aim of this study was to quantify Leishmania infantum parasites by real-time polymerase chain reaction (PCR) using specific DNA TaqMan probes and to compare the efficacy of detection of this technique with a PCR-enzyme-linked immunosorbent assay (ELISA) . For this purpose, spleen and liver samples from L . infantum-infected mice were collected during a 3-mo longitudinal study and analyzed by both methods . PCR-ELISA failed to quantify Leishmania spp . DNA in samples with very low or very high numbers of parasites . Real-time PCR was more sensitive than PCR-ELISA, detecting down to a single parasite, and enabled the parasite quantification over a wide, 5-log range . In summary, this study developed a method for absolute quantification of L . infantum parasites in infected organs using real-time TaqMan PCR.

J Parasitol, 2004 Oct, 90(5), 980 - 90
Microbial adhesion of Cryptosporidium parvum sporozoites: purification of an inhibitory lipid from bovine mucosa; Johnson JK et al.; Cryptosporidium parvum is a protozoan pathogen of humans and livestock worldwide . Its ability to infect a wide range of species raises questions as to the involvement of a specific host cell receptor for parasite-host recognition . To investigate the mechanism of parasite-host cell recognition, we have developed an in vitro cell suspension binding assay to investigate adhesion of C . parvum sporozoites to host cells . Morphologic features of binding events observed with this assay were identical to those described in natural infections . Glycoconjugates, Madin Darby bovine kidney (MDBK) cell fractions, and plasma membrane vesicles (PMVs) were screened for their ability to block binding of sporozoites to MDBK cells . Mucins, MDBK cell fractions, and PMVs exhibited dose-dependent inhibition of sporozoite binding . The major inhibitory fraction from MDBK cells was found to be insoluble in aqueous medium, nonsaponifiable, and lacking carbohydrate moieties, nitrogen, and phosphorus . Its inhibitory effect was resistant to heat, protease digestion, and glycosidase treatment, suggesting that the inhibitory activity is a lipid or a lipid-like component . The inhibitory activity was purified from MDBK cells, and in larger amounts from bovine small intestinal mucosa, by organic solvent extraction, semipreparative high-pressure liquid chromatography, and preparative high-performance thin-layer chromatography . Biochemical analyses, thin-layer chromatography staining techniques, mass spectrometry, and elemental analysis were used to partially characterize the purified lipid . These results indicate that a host intestinal lipid(s) or a lipid-like component(s) may play an important role in the early stages of host cell invasion by C . parvum.

J Biol Chem . 2004 Nov 23; {Epub ahead of print}
Collagenous transmembrane proteins: Recent insights into biology and pathology; Franzke CW et al.; This minireview will focus on a growing subgroup of collagens, the collagenous transmembrane proteins, which have dual functions as cell surface receptors or as matrix components . If their extracellular domains are cast off by limited proteolysis, they can begin to signal to cells as soluble molecules . Collagenous transmembrane proteins are widely expressed and are involved in cell adhesion, epithelial-mesenchymal interactions during morphogenesis, neuromuscular signaling, and host defense against microbial agents . Correspondingly, they are associated with genetic and acquired human diseases, e.g . epidermolysis bullosa, ectodermal dysplasias, bullous pemphigoid, or Alzheimer disease, and mouse models implicate them in the etiology of further pathological conditions.

J Clin Periodontol, 2004 Dec, 31(12), 1128 - 34
Comparative effects of different chlorhexidine mouth-rinse formulations on volatile sulphur compounds and salivary bacterial counts; Roldan S et al.; AIM: To compare five different commercial mouth rinses with chlorhexidine (CHX) with respect to their anti-halitosis effect and anti-microbial activity on salivary bacterial counts, following a standardised research protocol . And secondly, to validate the study model proposed in the evaluation of patients suffering from halitosis . PATIENTS AND METHODS: Ten volunteers, with a healthy oral status, were enrolled in a double-blind, cross-over design, using sterile saline as negative control and five CHX-containing mouth rinses: 0.12% CHX alone (CHX+NO), plus alcohol (CHX+ALC), plus 0.05% cetylpiridinium chloride (CHX+CPC), plus sodium fluoride (CHX+NaF), and 0.05% CHX plus 0.05% CPC, plus 0.14% zinc lactate (CHX+Zn) . The levels of whole-mouth volatile sulphur compounds (VSCs) were measured by means of a sulphide monitor at baseline, 1 and 5 h after rinsing with the assigned product . Baseline measurements also included an organoleptic assessment and the recording of the tongue-coating index . Aerobic and anaerobic salivary bacterial counts were also obtained by collecting unstimulated saliva samples at the same evaluation times, and processed by culturing techniques . Analysis of variance was used to evaluate whether significant differences existed among groups, at each evaluation point, or in changes between evaluations . RESULTS: No significant differences were detected at baseline, with VSC levels ranging between 190 and 227 parts per billion (p.p.b.) After rinsing, VSC levels were reduced with all products (except saline), after 1 h . Significant differences at 1 h were detected (p=0.04), corresponding to a lower amount of p.p.b . (109) in (CHX+Zn) as compared with the other groups (except CHX+NO) . At 5 h, VSC levels were lower for CHX+CPC and CHX+Zn (155 and 169, respectively), while the other groups showed levels higher than 220 p.p.b . With respect to aerobic salivary bacterial counts, CHX+CPC demonstrated the lowest percentage of survival (6% after 1 h and 18% after 5 h) . For anaerobic bacterial counts, again CHX+CPC demonstrated the lowest percentage of survival (10% at 1 h and 23% at 5 h), together with CHX+ALC (18% of survival at 5 h) . However, salivary counts and VSCs were only significantly correlated at baseline, but not after treatment . CONCLUSION: Important differences can be expected from different CHX formulations, in relation to both their anti-halitosis effect and anti-microbial activity in saliva . Formulations that combine CHX and CPC achieved the best results, and a formulation combining CHX with NaF resulted in the poorest.

J Environ Sci (China), 2004, 16(5), 772 - 8
Succession of aquatic microbial communities as a result of the water quality variations in continuous water; Wang RC et al.; The changes of structural and functional parameters of aquatic microbial communities in continuous water on campus of Tsinghua University, China are investigated, by polyurethane foam unit (PFU) method . The measured compositions of the communities include alga, protozoa, and some metazoa (such as rotifers) . The measured indicators of water quality include water temperature, pH value, dissolved oxygen (DO), potassium permanganate index (COD(Mn)), total nitrogen (TN), total phosphorus (TP) and chlorophyll-a (Chla) . The trophic level, expressed by the trophic level indices (TLIc), is assessed with analytic hierarchy process and principal component analysis (AHP-PCA) method . The changing trends of the structural and functional parameters of aquatic microbial communities, such as Margalef index of diversity (D), Shannon-weaver index of diversity (H), Heterotropy index (HI), number of species when the colonization gets equilibrium (S(eq)), colonizing speed constant (G) and time spent when 90 percent of S(eq) colonized in PFU (T(80%)), are also analyzed . The experimental results showed the succession of aquatic microbial communities along the water flow is consistent with the water quality changes, so the parameters of microbial community can reflect the changes of water quality from the ecological view.

Environ Toxicol Chem, 2004 Nov, 23(11), 2649 - 54
Persistence of selected organophosphate and carbamate insecticides in waters from a coastal watershed; Bondarenko S et al.; Organophosphate and carbamate compounds are among the most widely used pesticides . Contamination of surface water by these compounds is of concern because of potential toxicity to aquatic organisms, especially those at lower trophic levels . In this study we evaluated the persistence of diazinon, chlorpyrifos, malathion, and carbaryl in waters from various sites in the Newport Bay-San Diego Creek watershed in southern California (USA) . The persistence of diazinon and chlorpyrifos was much longer than that of malathion or carbaryl and was further prolonged in seawater . Microbial degradation contributed significantly to the dissipation of diazinon and chlorpyrifos in freshwater, but was inhibited in seawater, leading to increased persistence . In contrast, degradation of malathion and carbaryl was rapid and primarily abiotic . A greater temperature dependence was observed for carbaryl degradation in all waters and for diazinon degradation in freshwater . The interactions of pesticide persistence with water location, temperature, and type of pesticides suggest that site- and compound-specific information is needed when evaluating the overall ecotoxicological risks of pesticide pollution in a watershed . Because the persistence of diazinon and chlorpyrifos may increase significantly in seawater, mitigation should occur before the pesticides reach seawater . The relatively short persistence of these compounds in freshwater suggests that practices aimed at extending residence time (e.g., diversion to wetlands) may effectively reduce pesticide output to downstream water bodies.

Int Arch Occup Environ Health, 2004 Oct, 77(7), 461 - 70 Epub 2004 Oct.
Symptoms in relation to chemicals and dampness in newly built dwellings; Saijo Y et al.; OBJECTIVES: As the airtightness of dwellings has recently increased, problems associated with indoor air pollution and dampness have become important environmental health issues . The aim of this study was to clarify whether symptoms in residents living in newly built dwellings were related to chemicals and dampness . METHODS: Symptoms of 317 residents were surveyed by standardized questionnaires, and the concentrations of formaldehyde, acetaldehyde, and 17 volatile organic compounds (VOCs) in their homes were measured . Dampness (condensation on window panes and/or walls, and mold growth) was identified by questionnaires given to the householders or their partners . RESULTS: Some VOCs (toluene, butyl acetate, ethylbenzene, alpha-pinene, p-dichlorobenzene, nonanal, and xylene) were significantly related to the symptoms, and the sum of all VOCs (all identified VOCs) was significantly related to throat and respiratory symptoms {odds ratio (OR) for eye symptoms =2.4; 95% confidence interval (CI) 1.0-5.5}, although the concentrations of VOCs were relatively low . As for the dampness index, condensation on window panes and/or walls was related to all symptoms, and mold growth was related to all symptoms except skin, throat and respiratory and general symptoms . As the number of dampness signs increased, the ORs increased for the symptoms except general symptoms (OR for nose symptoms = 4.4, 95% CI 1.6-11.9) . CONCLUSION: Both VOCs and dampness were significantly related to symptoms . We should take measures to reduce the concentrations of VOCs, dampness and microbial growth in dwellings.

Postepy Hig Med Dosw (Online), 2004 Nov 22, 58, 398 - 404
{The mast cells phagocytose bacteria}; Rdzany RS et al.; In the last years there has been a growing number of reports concerning the role of mast cells in host defense against bacteria . The mast cell membrane is replete with many receptors/molecules, including those that promote the recognition and binding of bacteria . Mast cells exhibit two basic mechanisms of microbial recognition: opsonin-dependent (via Fc and C3 receptors) and opsonin-independent (via integrins, CD48 molecule and Toll-like receptors) . Moreover, mast cells phagocytose and kill adherent bacteria . Phagocytosis of bacteria results in the presentation of bacterial antigens for MHC class I to T cells.

J Exp Med, 2004 Dec 6, 200(11), 1419 - 26 Epub 2004 Dec 6.
A soluble form of the triggering receptor expressed on myeloid cells-1 modulates the inflammatory response in murine sepsis; Gibot S et al.; The triggering receptor expressed on myeloid cells (TREM)-1 is a recently discovered receptor expressed on the surface of neutrophils and a subset of monocytes . Engagement of TREM-1 has been reported to trigger the synthesis of proinflammatory cytokines in the presence of microbial products . Previously, we have identified a soluble form of TREM-1 (sTREM-1) and observed significant levels in serum samples from septic shock patients but not controls . Here, we investigated its putative role in the modulation of inflammation during sepsis . We observed that sTREM-1 was secreted by monocytes activated in vitro by LPS and in the serum of animals involved in an experimental model of septic shock . Both in vitro and in vivo, a synthetic peptide mimicking a short highly conserved domain of sTREM-1 appeared to attenuate cytokine production by human monocytes and protect septic animals from hyper-responsiveness and death . This peptide seemed to be efficient not only in preventing but also in down-modulating the deleterious effects of proinflammatory cytokines . These data suggest that in vivo modulation of TREM-1 by sTREM peptide might be a suitable therapeutic tool for the treatment of sepsis.

J Immunol, 2004 Dec 1, 173(11), 6873 - 81
Activation of virus-specific CD8+ T cells by lipopolysaccharide-induced IL-12 and IL-18; Raue HP et al.; Virus-specific T cells represent a hallmark of Ag-specific, adaptive immunity . However, some T cells also demonstrate innate functions, including non-Ag-specific IFN-gamma production in response to microbial products such as LPS or exposure to IL-12 and/or IL-18 . In these studies we examined LPS-induced cytokine responses of CD8(+) T cells directly ex vivo . Following acute viral infection, 70-80% of virus-specific T cells will produce IFN-gamma after exposure to LPS-induced cytokines, and neutralization experiments indicate that this is mediated almost entirely through production of IL-12 and IL-18 . Different combinations of these cytokines revealed that IL-12 decreases the threshold of T cell activation by IL-18, presenting a new perspective on IL-12/IL-18 synergy . Moreover, memory T cells demonstrate high IL-18R expression and respond effectively to the combination of IL-12 and IL-18, but cannot respond to IL-18 alone, even at high cytokine concentrations . This demonstrates that the synergy between IL-12 and IL-18 in triggering IFN-gamma production by memory T cells is not simply due to up-regulation of the surface receptor for IL-18, as shown previously with naive T cells . Together, these studies indicate how virus-specific T cells are able to bridge the gap between innate and adaptive immunity during unrelated microbial infections, while attempting to protect the host from cytokine-induced immunopathology and endotoxic shock.

Am J Respir Cell Mol Biol . 2004 Nov 19; {Epub ahead of print}
Moraxella catarrhalis-Infected Alveolar Epithelium Induced Monocyte Recruitment and Oxidative Burst; Rosseau S et al.; The recruitment of monocytes appears to be a crucial factor for inflammatory lung disease . Alveolar epithelial cells contribute to monocyte influx into the lung, but their impact on monocyte inflammatory capacity is not entirely clear . We thus analyzed the modulation of monocyte oxidative burst by A549 and isolated human alveolar epithelial cells . Epithelial infection with Moraxella catarrhalis induced monocyte adhesion, transepithelial migration, and superoxide generation, whereas stimulation with lipopolysaccharide, tumor necrosis factor-alpha, interleukin-1beta, or interferon-gamma induced adhesion or transmigration, but failed to initiate monocyte burst . The effect of microbial challenge was mimicked by phorbol myristate acetate and inhibited by the protein kinase C inhibitor bisindoylmaleimide . Furthermore, evidence for a role of platelet-activating factor-signaling in monocytes is presented . Monocyte burst was neither induced by supernatant nor affected by fixation of A549 cells, excluding the contribution of epithelium-derived soluble factors but emphasizing the mandatory role of intercellular contact . The employment of blocking antibodies, however, denied a role for the adhesion molecules intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, or CD11b/CD18 and CD49d/CD29 . In essence, infection of alveolar epithelial cells with M . catarrhalis might amplify the inflammatory capacity of invading monocytes eliciting their superoxide production . The epithelial response to this microbial challenge thus clearly differed from that to proinflammatory cytokines.

Water Res, 2004 Dec, 38(20), 4367 - 82
Behavior of trihalomethanes and haloacetic acids in a drinking water distribution system; Rodriguez MJ et al.; This investigation focused on the seasonal variation and spatial fate of chlorination disinfection by-products (CDBPs) in a drinking water distribution system located in a region where very significant seasonal variations in water temperature and surface water quality occur . The analysis of a large number of collected samples showed that the seasonal and geographical variations of both groups of CDBPs under study--trihalomethanes (THMs) and haloacetic acids (HAAs)--were particularly important in this region . THM levels in summer and fall were, on average, about five times higher than in winter, whereas average HAAs in spring were about four times higher than in winter . THMs increased and stabilized in the extremities of the distribution system, whereas HAAs begin to increase, and then decrease (mainly due to a reduction of dichloroacetic acid) . This decrease was significantly higher in warm waters than in cold waters, which led to the hypothesis of microbial degradation of HAAs as water approaches the system extremities . In fact, regression models for the occurrence of both CDBPs showed that the residence time of water was one important parameter in explaining the fate of both CDBPs . The spatio-temporal portrait of both groups of CDBPs that was generated demonstrates that, due to their high intra-seasonal changes, the calculation of average annual levels of these substances for compliance with regulations can vary widely . The results used in the portrait of CDBP behavior are also relevant in terms of exposure assessment for future epidemiological studies on human reproductive outcomes in the region.

Ann Ig, 2004 Jan-Apr, 16(1-2), 255 - 64
{Water resources destined for human consumption: qualitative monitoring}; Azara A et al.; The work is a synthesis of the qualitative monitoring on waters resources (specially surface water), that were potable and distributed as drinkable waters in north Sardinia between 1996-2002 . The obtained results point out: for the surface waters, recurrent problems of elevated turbidity, color, oxidability, microbial contamination and eutrofication; for the ground waters, lower levels of pollutants even if an high content of natural elements and substances that, for as of natural origin, could endanger and limit the use as drinkable water (sodium, chlorides, iron, manganese) . Drinking water plants have, largely, solved the main causes of ineffectiveness of treatment such as the demolition of the organic load and the formation of trihalomethanes but it as been found an high concentration of chlorites; occasionally, the values of ammonia, aluminum, iron, manganese and, rarely, trihalomethanes are out law in piped.

J Food Prot, 2004 Nov, 67(11), 2570 - 7
Home storage temperatures and consumer handling of refrigerated foods in Sweden; Marklinder IM et al.; The lack of data on consumer refrigeration temperatures and storage times limits our ability to assess and manage risks associated with microbial hazards . This study addressed these limitations by collecting data on temperatures and storage handling practices of chilled foods . Consumers from 102 households in Uppsala, Sweden, were instructed to purchase seven food items (minced meat, fresh herring fillets, soft cheese, milk, sliced cooked ham, vacuum-packed smoked salmon, and ready-to-eat salad) and to store them using their normal practices . They were interviewed the next day, and food temperatures were measured . In general, there were no significant relations between temperature and characteristics of the respondents (e.g., sex, age, education, age of the refrigerator) . Mean storage temperatures ranged from 6.2 degrees C for minced meat to 7.4 degrees C for ready-to-eat salad . Maximum temperatures ranged from 11.3 to 18.2 degrees C . Data were not significantly different from a normal distribution, except for ready-to-eat salad, although distributions other than the normal fitted data better in most cases . Five percent to 20% of the food items were stored at temperatures above 10 degrees C . Most respondents knew the recommended maximum temperature, but less than one fourth claimed to know the temperature in their own refrigerator . Practical considerations usually determined where food was stored . For products with a long shelf life, stated storage times were different for opened and unopened packages . The current situation might be improved if consumers could be persuaded to use a thermometer to keep track of refrigerator temperature.

J Food Prot, 2004 Nov, 67(11), 2375 - 80
Efficacy of sanitizers to inactivate Escherichia coli O157:H7 on fresh-cut carrot shreds under simulated process water conditions; Gonzalez RJ et al.; Chlorine is widely used as a sanitizer to maintain the microbial quality and safety of fresh-cut produce; however, chlorine treatment lacks efficacy on pathogen reduction, especially when the fresh-cut processing water contains heavy organic loads . A more efficacious sanitizer that can tolerate the commercial processing conditions is needed to maintain microbial safety of fresh-cut produce . This study evaluated the efficacy of Escherichia coli O157:H7 reduction on fresh-cut carrots using new and traditional sanitizers with tap water and fresh-cut processing water scenarios . Fresh-cut carrot shreds inoculated with E . coli O157:H7 were washed in sanitizer solutions including 200 ppm chlorine, citric acid-based sanitizer (Pro-San), 80 ppm peroxyacetic acid-based sanitizer (Tsunami 100), and 1,000 ppm acidified sodium chlorite (SANOVA) prepared in fresh tap water or simulated processing water with a chemical oxygen demand level of approximately 3,500 mg/liter . Samples were packaged and stored at 5 degrees C . Microbial analyses performed at days 0, 7, and 14 indicate that the organic load in the process water significantly affected the efficacy of chlorine on pathogen removal and was especially evident on samples tested during storage . Acidified sodium chlorite provided a strong pathogen reduction even under process water conditions with up to a 5.25-log reduction when compared with the no-wash control . E . coli O157:H7 was not recovered on acidified sodium chlorite-treated samples during the entire 14 days of storage, even following an enrichment step . These results suggest that acidified sodium chlorite holds considerable promise as an alternative sanitizer of fresh-cut produce.

Environ Technol, 2004 Oct, 25(10), 1107 - 14
Startup of an anaerobic fluidized bed reactor for landfill leachate treatment; Gulsen H et al.; Anaerobic treatment of the young landfill leachate in a pilot-scale fluidized bed reactor was performed to observe the effects of startup on COD removal, biogas production and biomass attachment . During the experiments, the organic loading rate (OLR) was increased gradually from 2.5 to 27 g COD l(-1) day(-1) with varying the feed rate and the hydraulic retention time in the range of 5.5-13 1 day(-1) and 1-2.35 days, respectively . The anaerobic fluidized bed reactor (AFBR) showed an excellent performance and attained steady state conditions with COD removal of 90% after 80 days . Biogas production in the reactor continuously increased and its average yield (Y(gas)) was defined as 0.531 biogas g(-1) COD(rem)(-1) with a correlation of 99% . Greater microbial attachment was found at the increased loading rates and the attached biomass concentration (X(attach)) consisted of about 90% of the total biomass concentration . In addition, the substrate utilization rate indicated a linear increase as a function of time during the process.

Clin Exp Dermatol, 2004 Nov, 29(6), 584 - 8
Significance of the cytokine interferon gamma in clinical dermatology; Grassegger A et al.; Interferon (IFN)-gamma is considered a key cytokine of innate and adaptive immunity playing pivotal roles in host defence against microbial pathogens and tumours, and exerts profound antiproliferative and antifibrotic effects . In this review we discuss applications and perspectives of IFN-gamma in clinical dermatology, such as papillomavirus and bacterial infections, tumours, atopic dermatitis, and fibrotic conditions such as scleroderma and postradiation fibrosis . Moreover, we give a summary of the pharmacologic properties including main side effects and potential risk factors of IFN-gamma therapy . Although former enthusiasm for IFN-gamma (e.g . in atopic dermatitis) has subsided, this cytokine might remain a promising tool (and target) in clinical dermatology, due to its central immunobiologic functions, better characterization of its kinetics in diseases facilitating optimized treatment schedules, and successful applications in fibrotic conditions such as scleroderma, idiopathic pulmonary and skin postradiation fibrosis.

Anaesthesia, 2004 Dec, 59(12), 1221 - 3
Microbial keratitis in ITU staff: an occupational hazard?
Ezra DG, Goyal S, Moosavi R, Millar M, Laganowski HC, Moore AT.
Microbial keratitis is a potentially blinding corneal infection; the infection may progress rapidly if untreated and result in corneal perforation . The breakdown of the innate ocular defences is known to predispose to corneal infection . We present three cases of microbial keratitis in intensive therapy unit (ITU) staff each of whom had compromised corneal immunity . Anaesthetists and nurses regularly perform high-risk procedures, such as tracheal suctioning, which have been known to cause ocular infections by aerosol inoculation . We suggest that although the absolute risk of infection is low, susceptible individuals may be at increased risk of corneal infection from exposure to potentially pathogenic organisms in such environments . We stress the need for ITU staff, particularly those who wear contact lenses, to wear eye protection when performing procedures likely to cause infected aerosols.

Bioorg Med Chem Lett, 2004 Dec 20, 14(24), 6035 - 40
Design, synthesis and anti-microbial activity of 1H-pyrazole carboxylates; Sridhar R et al.; In a SAR study, we have synthesized a few 1H-pyrazole carboxylate related microbicides using Vilsmeier reagent . The anti-microbial screening results of 1H-pyrazole-3-carboxylate are reported here for the first time . The effect of 1H-pyrazole carboxylates on the mycelial growth of plant pathogenic fungi is revealed . The first X-ray structure in the family of microbicidal 1H-pyrazole-4-carboxylates is presented.

J Appl Microbiol, 2004, 97(6), 1132 - 9
In situ quantification of biocide efficacy using GFP transformed Aureobasidium pullulans; Sabev HA et al.; AIMS: To develop a real-time in situ method to quantify loss of viability of Aureobasidium pullulans PRAFS8 cells attached to plasticized polyvinyl chloride (pPVC) with incorporated biocides, and to use the method to compare biocide efficacy in situ . METHODS AND RESULTS: A . pullulans PRAFS8, transformed with green fluorescent protein (GFP), was used to quantify the efficacy of a range of biocides incorporated into pPVC . Experimentally, it was found that a density of 1.53 x 10(6) yeast cells per cm(2) of pPVC was optimal as increasing the density of the yeast cells to 6.12 x 10(6) cm(-2) attached to pPVC containing the biocide 2-n-octyl-4-isothiazolin-3-one (OIT) decreased the rate of fluorescence loss . A strong positive correlation between fluorescence and viable yeast cell number was observed and fluorescence was used as a direct indicator of cell viability . The effectiveness of five commercial biocides, commonly incorporated into pPVC at their in-use concentrations, was tested against yeast cells attached to the pPVC surface . The loss of fluorescence and hence viability in situ was quantified using image analysis . The biocides N-(trichloromethylthio) phthalimide (NCMP), 10,10'-oxybisphenoxarsine (OBPA), OIT and 2,3,5,6-tetrachloro-4-(methylsulphonyl) pyridine (TCMP) caused complete loss of fluorescence within 30-50 h . In contrast the biocide dichloro-octyl-isothiazoline caused only 55 +/- 15% fluorescence loss after 50 h . Starvation of the yeast cells in suspension for 24 h prior to attachment reduced their initial sensitivity to OBPA, NCMP, OIT and TCMP by 15-20%, but eventually the fluorescence was also completely lost . CONCLUSIONS: The use of A . pullulans expressing cytosolic GFP enables the in situ quantification of loss of viability when cells are attached to pPVC with incorporated biocides . SIGNIFICANCE AND IMPACT OF THE STUDY: GFP fluorescence was used as a real-time indicator of cell viability and thus can be applied for direct quantification of the effectiveness of a broad range of biocides, incorporated into the polymer mass and used to protect a variety of plastics or other materials from microbial growth.

Plant J, 2004 Dec, 40(5), 633 - 46
Host and non-host pathogens elicit different jasmonate/ethylene responses in Arabidopsis; Zimmerli L et al.; Arabidopsis does not support the growth and asexual reproduction of the barley pathogen, Blumeria graminis f . sp . hordei Bgh) . A majority of germlings fail to penetrate the epidermal cell wall and papillae . To gain additional insight into this interaction, we determined whether the salicylic acid (SA) or jasmonate (JA)/ethylene (ET) defence pathways played a role in blocking barley powdery mildew infections . Only the eds1 mutant and NahG transgenics supported a modest increase in penetration success by the barley powdery mildew . We also compared the global gene expression patterns of Arabidopsis inoculated with the non-host barley powdery mildew to those inoculated with a virulent, host powdery mildew, Erysiphe cichoracearum . Genes repressed by inoculations with non-host and host powdery mildews relative to non-inoculated control plants accounted for two-thirds of the differentially expressed genes . A majority of these genes encoded components of photosynthesis and general metabolism . Consistent with this observation, Arabidopsis growth was inhibited following inoculation with Bgh, suggesting a shift in resource allocation from growth to defence . A number of defence-associated genes were induced during both interactions . These genes likely are components of basal defence responses, which do not effectively block host powdery mildew infections . In addition, genes encoding defensins, anti-microbial peptides whose expression is under the control of the JA/ET signalling pathway, were induced exclusively by non-host pathogens . Ectopic activation of JA/ET signalling protected Arabidopsis against two biotrophic host pathogens . Taken together, these data suggest that biotrophic host pathogens must either suppress or fail to elicit the JA/ET signal transduction pathway.

Microb Ecol, 2004 Aug, 48(2), 218 - 29 Epub 2004 Jun 10.
Microbial community structure and oxidative enzyme activity in nitrogen-amended north temperate forest soils; Gallo M et al.; Large regions of temperate forest are subject to elevated atmospheric nitrogen (N) deposition which can affect soil organic matter dynamics by altering mass loss rates, soil respiration, and dissolved organic matter production . At present there is no general model that links these responses to changes in the organization and operation of microbial decomposer communities . Toward that end, we studied the response of litter and soil microbial communities to high levels of N amendment (30 and 80 kg ha(-1) yr(-1)) in three types of northern temperate forest: sugar maple/basswood (SMBW), sugar maple/red oak (SMRO), and white oak/black oak (WOBO) . We measured the activity of extracellular enzymes (EEA) involved directly in the oxidation of lignin and humus (phenol oxidase, peroxidase), and indirectly, through the production of hydrogen peroxide (glucose oxidase, glyoxal oxidase) . Community composition was analyzed by extracting and quantifying phospholipid fatty acids (PLFA) from soils . Litter EEA responses at SMBW sites diverged from those at oak-bearing sites (SMRO, BOWO), but the changes were not statistically significant . For soil, EEA responses were consistent across forests types: phenol oxidase and peroxidase activities declined as a function of N dose (33-73% and 5-41%, respectively, depending on forest type); glucose oxidase and glyoxal oxidase activities increased (200-400% and 150-300%, respectively, depending on forest type) . Principal component analysis (PCA) ordinated forest types and treatment responses along two axes; factor 1 (44% of variance) was associated with phenol oxidase and peroxidase activities, factor 2 (31%) with glucose oxidase . Microbial biomass did not respond to N treatment, but nine of the 23 PLFA that formed >1 mol% of total biomass showed statistically significant treatment responses . PCA ordinated forest types and treatment responses along three axes (36%, 26%, 12% of variance) . EEA factors 1 and 2 correlated negatively with PLFA factor 1 ( r = -0.20 and -0.35, respectively, n = 108) and positively with PLFA factor 3 ( r = +0.36 and +0.20, respectively, n = 108) . In general, EEA responses were more strongly tied to changes in bacterial PLFA than to changes in fungal PLFA . Collectively, our data suggests that N inhibition of oxidative activity involves more than the repression of ligninase expression by white-rot basidiomycetes.

Microb Ecol, 2004 Aug, 48(2), 191 - 9 Epub 2004 Jun 10.
Bacterial density and community structure associated with aggregate size fractions of soil-feeding termite mounds; Fall S et al.; The building and foraging activities of termites are known to modify soil characteristics such as the heterogeneity . In tropical savannas the impact of the activity of soil-feeding termites ( Cubitermes niokoloensis) has been shown to affect the properties of the soil at the aggregate level by creating new soil microenvironments (aggregate size fractions) {13} . These changes were investigated in greater depth by looking at the microbial density (AODC) and the genetic structure (automated rRNA intergenic spacer analysis: ARISA) of the communities in the different aggregate size fractions (i.e., coarse sand, fine sand, coarse silt, fine silt, and dispersible clays) separated from compartments (internal and external wall) of three Cubitermes niokoloensis mounds . The bacterial density of the mounds was significantly higher (1.5 to 3 times) than that of the surrounding soil . Within the aggregate size fractions, the termite building activity resulted in a significant increase in bacterial density within the coarser fractions (>20 mum) . Multivariate analysis of the ARISA profiles revealed that the bacterial genetic structures of unfractionated soil and soil aggregate size fractions of the three mounds was noticeably different from the savanna soil used as a reference . Moreover, the microbial community associated with the different microenvironments in the three termite mounds revealed three distinct clusters formed by the aggregate size fractions of each mound . Except for the 2-20 mum fraction, these results suggest that the mound microbial genetic structure is more dependent upon microbial pool affiliation (the termite mound) than on the soil location (aggregate size fraction) . The causes of the specificity of the microbial community structure of termite mound aggregate size fractions are discussed.

Crit Care Med, 2004 Nov, 32(11 Suppl), S513 - 26
Source control in the management of severe sepsis and septic shock: an evidence-based review; Marshall JC et al.; OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for source control in the management of severe sepsis and septic shock that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and to improve outcome in severe sepsis . DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee . METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum . We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade . Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al . on p . S591 . CONCLUSION: Source control represents a key component of success in therapy of sepsis . It includes drainage of infected fluids, debridement of infected soft tissues, removal of infected devices or foreign bodies, and finally, definite measures to correct anatomic derangement resulting in ongoing microbial contamination and to restore optimal function . Although highly logical, since source control is the best way to reduce quickly the bacterial inoculum, most recommendations are, however, graded as D or E due to the difficulty to perform appropriate randomized clinical trials in this respect . Appropriate source control should be part of the systematic checklist we have to keep in mind in setting up the therapeutic strategy in sepsis.

Endocrinology, 2005 Feb, 146(2), 835 - 44 Epub 2004 Nov 11.
Reevaluating thyrotropin receptor-induced mouse models of graves' disease and ophthalmopathy; Baker G et al.; We aimed to establish and extend the characterization of murine models of thyroiditis and Graves' ophthalmopathy, induced by transfer of TSH receptor (TSHR) primed T cells . Experiments were performed in a different animal unit but using female BALB/cbyJico mice from the same supplier as previously . We report our findings together with a reevaluation of the earlier studies . In the first experiment, genetic immunization or TSHR fusion protein induced TSHR antibodies in all nine mice . Some of the antibodies functioned as thyroid-stimulating antibodies and/or TSH binding inhibiting Igs with two of seven mice having elevated T(4) . Thyroiditis and orbital changes were absent . Splenocyte transfer induced no immune response in naive BALB/cbyJico recipients . Subsequently genetic immunization or fusion protein-treated mice were maintained in either local or Brussels conditions (water, chow, and bedding) . TSHR antibodies were induced in nine of nine Brussels (with decreased T(4) in one of nine) but five of nine local mice . No thyroiditis or orbital changes were induced, but misleading fixation artefacts in extraocular muscles were noted . Nonspecific in vitro stimulation induced more CD-4+/IL-4+ cells in Brussels maintained . TSHR stimulation produced a significant increase in IL-4 secretion in six of nine local but one of seven Brussels mice . Thyroids from many TSHR-treated and control mice contained ectopic thymus . Our results confirm that thyroiditis is required for disease transfer but indicate the heterogeneity in TSHR-induced immune response in an inbred strain . Ectopic thymus can masquerade as thyroiditis, and care is required to avoid muscle artefacts . Because neither animal unit is pathogen free, microbial environment may contribute to determining TSHR-induced responses.

Trends Endocrinol Metab, 2004 Dec, 15(10), 500 - 5
Role of PPARgamma in macrophage biology and atherosclerosis; Zhang L et al.; Macrophages carry out key functions by defending a host from microbial invaders and by clearing endogenous cellular debris . Molecules that are essential for the recognition, phagocytosis and clearance of pathogens also mediate the uptake and degradation of pathogenic lipoproteins . During atherogenesis, for example, scavenging trapped lipoproteins leads to the formation of foam cells and subsequently the activation of these lipid-laden macrophages . Although they are initially clinically silent, these fatty streaks evolve into complex inflammatory plaques that cause significant morbidity and mortality . Thus, interventions that decrease foam cell formation and reduce the inflammatory response of macrophages could become effective therapies for coronary artery disease . Thiazolidinediones (TZDs) might be developed as anti-atherogenic agents on the basis of their actions as ligands for peroxisome proliferator-activated receptor-gamma (PPARgamma).

Horm Res, 2004, 62 Suppl 3, 98 - 103
Liquid growth hormone: preservatives and buffers; Kappelgaard AM et al.; Growth hormone (GH) treatment is a successful medical therapy for children and adults with GH deficiency as well as for growth retardation due to chronic renal disease, Turner syndrome and in children born small for gestational age . For all of these conditions, treatment is long term and patients receive daily subcutaneous injections of GH for many years . Patient compliance is therefore of critical importance to ensure treatment benefit . One of the major factors influencing compliance is injection pain . Besides the injection device used, pain perception and local tissue reaction following injection are dependent on the preservative used in the formulation and the concentration of GH . Injection pain may also be related to the buffer substance and injection volume . A liquid formulation of GH, Norditropi SimpleXx, has been developed that dispenses with the need for reconstitution before administration . The formulation uses phenol (3 mg/ml) as a preservative (to protect product from microbial degradation or contamination) and histidine as a buffer . Alternative preservatives used in other GH formulations include m-cresol (9 mg/ml) and benzyl alcohol (3-9 mg/ml) . Buffering agents include citrate and phosphate . Phenol has been successfully used as a preservative in drug formulations for more than 50 years and is considered a safe and effective agent which complies with strict international requirements for preservatives in drug formulations . In toxicological studies, no or only mild local reactions have been observed following subcutaneous administration of phenol (7.5 mg/ml), m-cresol (3-4 mg/ml) and benzyl alcohol (9 mg/ml) . No general toxicity reactions were observed after subcutaneous administration of these agents . Clinical evaluation of the preservatives and buffers used in Norditropin SimpleXx showed that pain perception was similar between formulations containing phenol and benzyl alcohol, whereas m-cresol was associated with more painful injections than benzyl alcohol . Furthermore, patients reported more pain following injection of a citrate-buffered solution than after a histidine-buffered solution . More pain was also reported following large volume injections and following injections with solutions containing high protein concentrations . In summary, optimization of the preservative and buffer content of a liquid GH formulation may reduce injection pain and lead to improved patient compliance . 2004 S . Karger AG, Basel.

Immunity, 2004 Nov, 21(5), 733 - 41
Toll-dependent control mechanisms of CD4 T cell activation; Pasare C et al.; Toll-like receptors (TLRs) detect microbial infection and play an essential role in the induction of innate and adaptive immune responses . The mechanisms of TLR-mediated control of adaptive immunity are not yet fully understood . Induction of dendritic cell (DC) maturation is essential for activation of naive T cells . Here, we demonstrate that TLR-induced DC maturation and migration to the lymph nodes, in the absence of TLR-induced inflammatory cytokines, are not sufficient for T cell activation in vivo . We show that transient depletion of regulatory T (Tr) cells recovers the primary CD4 T cells response in MyD88-deficient mice, demonstrating that a major mechanism of TLR-mediated activation of T cell responses is the blocking of suppression by regulatory T cells . In addition we show that a TLR-induced signal(s) is required for memory CD4 T cell differentiation, but not for activation of memory T cells.

Curr Infect Dis Rep, 2004 Dec, 6(6), 450 - 456
Vaginal Immunity in Bacterial Vaginosis; Cauci S; Vaginal immunity in response to microbial perturbation is still poorly understood and may be crucial for protection from adverse outcomes associated with bacterial vaginosis (BV) . BV is the most prevalent vaginal disorder in adult women worldwide . However, its pathogenesis is still elusive . In BV-positive women, inflammatory signs are scant--approximately 50% of women are asymptomatic . The number of vaginal neutrophils in the BV-positive patient is not increased with respect to healthy women . In contrast, vaginal interleukin (IL)-1beta levels are largely increased . Recent findings indicate that microbial hydrolytic enzymes could be responsible for dampening the expected proinflammatory response cascade after IL-1beta increase . In other words, BV causes a large increase of vaginal IL-1beta, which is not paralleled by an increase of IL-8 levels, suggesting that BV-associated factors specifically dampen IL-8 . The impairment of IL-8 increase may explain the absence of neutrophil increase in most women exposed to a massive abnormal anaerobic vaginal colonization (BV) . Among BV-positive women, vaginal innate immunity is strongly correlated to a specific adaptive immune response: the immunoglobulin A (IgA) against the hemolysin produced by Gardnerella vaginalis (anti-Gvh IgA), which is the main bacterium present in BV . High anti-Gvh IgA levels are protective for adverse pregnancy outcomes . However, an exaggerated inflammatory response, mainly attributed to genetic polymorphisms, is also implicated in BV-associated adverse outcomes.

Appl Microbiol Biotechnol, 2005 Jan, 66(4), 352 - 5 Epub 2004 Nov 05.
Towards electronic paper displays made from microbial cellulose; Shah J et al.; Cellulose (in the form of printed paper) has always been the prime medium for displaying information in our society and is far better than the various existing display technologies . This is because of its high reflectivity, contrast, low cost and flexibility . There is a major initiative to push for a dynamic display technology that emulates paper (popularly known as "electronic paper") . We have successfully demonstrated the proof of the concept of developing a dynamic display on cellulose . To the best of our knowledge, this is the first significant effort to achieve an electronic display using bacterial cellulose . First, bacterial cellulose is synthesized in a culture of Acetobacter xylinum in standard glucose-rich medium . The bacterial cellulose membrane thus formed (not pulp) is dimensionally stable, has a paper-like appearance and has a unique microfibrillar nanostructure . The technique then involves first making the cellulose an electrically conducting (or semi-conducting) sheet by depositing ions around the microfibrils to provide conducting pathways and then immobilizing electrochromic dyes within the microstructure . The whole system is then cased between transparent electrodes, and upon application of switching potentials (2-5 V) a reversible color change can be demonstrated down to a standard pixel-sized area (ca . 100 mum(2)) . Using a standard back-plane or in-plane drive circuit, a high-resolution dynamic display device using cellulose as substrate can be constructed . The major advantages of such a device are its high paper-like reflectivity, flexibility, contrast and biodegradability . The device has the potential to be extended to various applications, such as e-book tablets, e-newspapers, dynamic wall papers, rewritable maps and learning tools.

J Environ Qual, 2004 Nov-Dec, 33(6), 2157 - 64
Effect of combined application of methyl isothiocyanate and chloropicrin on their transformation; Zheng W et al.; Combining several soil fumigants to increase the broad spectrum of pest control is a common fumigation practice in current production agriculture . In this study, we investigated the effect of combined application of chloropicrin and methyl isothiocyanate (MITC) on their transformations and persistence in the environment . In aqueous solution, no direct reaction between MITC and chloropicrin occurred and relatively slow rates of hydrolysis of these compounds were observed in aquatic environments free of suspended solids . The transformation of chloropicrin, however, was accelerated in aqueous solution with MITC because of a reduction reaction with bisulfide (HS(-)), which is a by-product of MITC hydrolysis . In soil, when fumigants were applied simultaneously, the degradation of MITC was suppressed under the bi-fumigant application due to the inhibition of soil microbial activity and a possible abiotic competition with chloropicrin for a limited number of reaction sites on the surface of soil particles . However, the degradation rate of chloropicrin was significantly enhanced in the bi-fumigant soil system, which was primarily attributed to the reaction of chloropicrin and HS(-) . Two sequential application approaches were developed to investigate the feasibility of the combined application of metam sodium (parent compound of MITC) and chloropicrin in soil and assess their potential effects on environmental fate . For both application sequences, the degradation of chloropicrin was accelerated and that of MITC, as a major breakdown product of metam sodium, was inhibited in soil.

J Anim Sci, 2004 Dec, 82(12), 3567 - 76
Effects of supplemental protein type on intake, nitrogen balance, and site, and extent of digestion in whiteface wethers consuming low-quality grass hay; Salisbury MW et al.; Two experiments were conducted to determine the effects of supplementing ruminally degradable intake protein (DIP) or ruminally undegradable intake protein (UIP) on N balance (Exp . 1; n = 6 wethers; initial BW = 48.7 +/- 4.6 kg) and site and extent of digestion (Exp . 2; n = 5 wethers; initial BW = 36.9 +/- 3.1 kg) in whiteface wethers consuming (as-fed basis) 69% blue grama and 31% love grass hay (mixture = 7.5% CP, 73.0% NDF, 36.0% ADF {DM basis}) . Treatments were 1) no supplement (Control), 2) a supplement (219 g/d, as-fed basis) low in UIP (70 g/d of CP; 24.8 g/d of UIP), and 3) a supplement (219 g/d, as-fed basis) high in UIP (70 g/d of CP; 37.1 g/d of UIP) . Both experiments were replicated 3 x 3 Latin square designs, with identical feeding and supplementation . Wethers had ad libitum access to the forage mixture and fresh water, and received supplement once daily . In Exp.1, forage intake (percentage of BW) was greatest (P = 0.04) for control, but total DMI (g/d) was greatest (P = 0.05) for lambs consuming supplement . Apparent total-tract OM digestibility was numerically greater (P = 0.11) for supplemented wethers than for controls, whereas total-tract ADF digestibility tended (P = 0.08) to be greater for control wethers . Lambs fed supplements consumed and retained more (P < or = 0.01) N (% of N intake) compared with controls, but no difference (P = 0.22) was observed between low and high UIP treatments . Similar to Exp . 1, forage intake (percentage of BW) tended (P = 0.06) to be greater for control than for supplemented wethers in Exp . 2 . Ruminal NDF digestibility was 16.3% greater (P = 0.02) for supplemented wethers than for controls . Postruminal NDF and N digestibilities were greatest (P < or = 0.03) for controls, but apparent OM digestibility did not differ among treatments at all sites . Duodenal N flow was greatest (P = 0.05) for high UIP and least for control wethers . Nonmicrobial N flow was greater (P = 0.02) for high UIP compared with low UIP or controls . Control wethers had greater (P = 0.05) microbial efficiency . Ruminal ammonia concentration tended (P = 0.08) to be greatest for wethers fed low UIP and least for controls, with high-UIP wethers having intermediate ammonia concentrations . Results from these experiments suggest that in lambs fed low-quality forage there was no difference in apparent total-tract digestion or N balance (percentage of N intake) between lambs fed supplements that had the same CP but differed in the proportion of UIP and DIP; however, supplementing protein (regardless of UIP:DIP ratio) to wethers consuming low-quality forage increased N balance.

J Med Chem, 2004 Nov 18, 47(24), 5860 - 71
Fluorinated phenylcyclopropylamines . 2 . Effects of aromatic ring substitution and of absolute configuration on inhibition of microbial tyramine oxidase; Rosen TC et al.; A series of para-substituted diastereopure cis- and trans-2-fluoro-2-arylcyclopropylamines were synthesized and these were investigated as inhibitors of microbial tyramine oxidase from Arthrobacter sp . All compounds were shown to be competitive inhibitors of this enzyme . The nature of the para-substituents in the more potent trans-isomer (cis-relationship between fluorine and the amino group) of 2-fluoro-2-arylcyclopropylamine influenced the inhibitory potency in a consistent fashion . Thus, electron-withdrawing groups (F, Cl) slightly decreased the activity, while the methyl group (+ I substituent) increased the activity by a factor of ca . 7 compared to trans-2-fluoro-2-phenylcyclopropylamine and by a factor of 90 compared to tranylcypromine . Activity also was strongly dependent on the absolute configuration . The (1S,2S)-enantiomer of 2-fluoro-2-phenylcyclopropylamine was an excellent inhibitor of tyramine oxidase whereas the (1R,2R)-enantiomer was essentially devoid of activity.

Water Sci Technol, 2004, 50(6), 9 - 15
Biological removal of NOx from flue gas; Kumaraswamy R et al.; BioDeNOx is a novel integrated physico-chemical and biological process for the removal of nitrogen oxides (NOx) from flue gas . Due to the high temperature of flue gas the process is performed at a temperature between 50-55 degrees C . Flue gas containing CO2, O2, SO2 and NOx, is purged through Fe(II)EDTA2- containing liquid . The Fe(II)EDTA2- complex effectively binds the NOx; the bound NOx is converted into N2 in a complex reaction sequence . In this paper an overview of the potential microbial reactions in the BioDeNOx process is discussed . It is evident that though the process looks simple, due to the large number of parallel potential reactions and serial microbial conversions, it is much more complex . There is a need for a detailed investigation in order to properly understand and optimise the process.

Skin Res Technol, 2004 Nov, 10(4), 283 - 6
Assessment of spray application of Saint GERVAIS water effects on skin wettability by contact angle measurement comparison with bidistilled water; Elkhyat A et al.; BACKGROUND: The skin is responsible for protecting the body from physical, chemical and microbial injuries . The stratum corneum is the top layer of the epidermis and it plays a key role in helping to contain moisture . When the skin becomes damaged, its ability to perform these functions is compromised . Dry skin is a common form of skin damage.Contact angle 0 between a surface and water is a good indicator of hydrophobic or hydrophilic tendency of surfaces . OBJECTIVE: To quantify the influence of spray application of an isotonic mineral water (Saint GERVAIS water) on the hydrophobic tendency of dry skin and to compare spreading of both types of water on the skin through 0 measurement of each water drop on the skin before and after mineral spray application . MATERIALS AND METHODS: Contact angles theta between skin surface and the two types of water (bidistilled and mineral water) were measured before and after application of mineral water by a system allowing the measurement of theta in vivo and in vitro . The cutaneous hydration, the temperature and pH were also measured before and after mineral water application . RESULTS: As for the application of mineral water, the cutaneous pH significantly increased so the strong skin acidity decreased . The cutaneous temperature was modified (a decrease of 2 degrees C was noted) and the cutaneous hydration was also increased (+47AU) . The hydro-phobic tendency of the dry skin tested in this study was strongly decreased after mineral water application . This parameter was illustrated by a decrease of contact angle theta value . CONCLUSION: Saint GERVAIS water application allowed a regulation of skin pH, a fresh sensation on the skin and an increase in the cutaneous hydration . Moreover, the contact angle measurement with bidistilled water illustrated the increased wettability of the skin.

Mycol Res, 2004 Oct, 108(Pt 10), 1227 - 33
Distribution and estimation of anaerobic zoosporic fungi along the digestive tracts of sheep; Rezaeian M et al.; The status of anaerobic zoosporic (Chytridiomycota) fungi along the entire digestive tract of sheep was assessed both analytically and microscopically . Digest samples were taken from different segments of the digestive tracts of three newly killed sheep that previously had been used in experimental dietary studies . These digest samples were tested for the presence of rumen fungi by assessing the recovery of live fungi from the samples, direct observation of digested plant remains under the scanning electron microscope (SEM), and using a chitin assay as an estimation of fungal biomass . Live anaerobic fungi were recovered from the abomasum, small and large intestine, caecum and faeces of sheep, but not from the digest samples of rumen and omasum . However, SEM examination of the samples confirmed the presence of fungal structures from all of these organs . In the large intestine and caecum samples the observed sporangial structures were rounded and showed conspicuous surface pitting . Results of the chitin assay indicated that the anaerobic fungi might account for up to 20% of the total microbial biomass in the rumen of sheep . The results of this study support the view that anaerobic fungi may be present as a resistant stage in the lower reaches of the digestive tract.

Mass Spectrom Rev . 2004 Nov 8; {Epub ahead of print}
A review of current applications of mass spectrometry for biomarker/molecular tracer elucidations; Simoneit BR; I.Introduction000A . Biomarker Definitions000 1 . Higher-Plant Biomarkers000 a . Lipids000 b . Terpenoids000 2 . Microbial Biomarkers000B . Anthropogenic Tracer Compounds000II.Mass Spectrometry of Biomarkers000A . Common Mass Spectrometry Methods000 1 . Gas Chromatography-Mass Spectrometry000 2 . Gas Chromatography-Isotope Ratio Mass Spectrometry000 3 . Tandem Mass Spectrometry000B . Other Mass Spectrometry Methods000 1 . High-Temperature Gas Chromatography-Mass Spectrometry000 2 . Gas Chromatography-High-Resolution Mass Spectrometry000 3 . Pyrolysis Gas Chromatography-Mass Spectrometry000 4 . High-Pressure Liquid Chromatography-Mass Spectrometry000 5 . Time-of-Flight Mass Spectrometry000III.Applications of Mass Spectrometry in Biomarker Research000A . Geochemistry000 1 . Biomarkers in the Fossil Record000 2 . Petroleum Biomarkers000 a . Immature Precursors000 b . Triterpane Maturation000 c . Sterane Maturation000 3 . Aromatic (Coal) Biomarkers000 4 . Sulfurized Biomarkers000 5 . Novel Biomarkers000B . Environmental Chemistry000 1 . Natural Product and Fossil Fuel Biomarkers000 2 . Anthropogenic Compounds000C . Archeological and Forensic Chemistry000 1 . Biomarkers in Archeology000 2 . Use of Biomarkers in Forensics000D . Organic Cosmochemistry000 1 . Biomarkers in Extraterrestrial Samples000 2 . Biomarkers of Past Microbial Ecosystems000 3 . Biomarkers of Extant Microbial Ecosystems000 4 . Abiogenic Organic Compounds000 5 . Non-Biomarkers000IV.Prognosis000Acknowledgments000Appendix: Key Chemical Structures Cited in the Text000References000Mass spectrometry, especially coupled with gas chromatography or tandem, is the analytical method of choice in elucidations of biomarker compounds present in organic mixtures extracted from geological, environmental, or biological samples . This review describes the biomarker concept; i.e., the precursor natural products to the geological/environmental derivatives, and their application as tracers in the geosphere and ambient environment . The mass spectrometric methods currently utilized for such analyses are reviewed, and typical examples of applications are described with a general key to the literature . (c) 2004 Wiley Periodicals, Inc, Mass Spec Rev.

Proc Natl Acad Sci U S A, 2004 Nov 16, 101(46), 16246 - 50 Epub 2004 Nov 16.
Evolving protein functional diversity in new genes of Drosophila; Zhang J et al.; The mechanism by which protein functional diversity expands is an important evolutionary issue . Studies of recently evolved chimeric genes permit direct investigation of the origin of new protein functions before they become obscured by subsequent evolution . Found in several African Drosophila species, jingwei (jgw), a recently evolved gene with a domain derived from the still extant short-chain alcohol dehydrogenase (ADH) through retroposition, provides an opportunity to examine this previously undescribed process directly . We expressed JGW proteins in a microbial expression system and, after purification, investigated their enzymatic properties . We found that, unexpectedly, positive Darwinian selection for amino acid replacements outside the active site of JGW produced a novel dehydrogenase with altered substrate specificity compared with the ancestral ADH . Instead of detoxifying and assimilating ethanol like its Adh parental gene, we observe that JGW efficiently utilizes long-chain primary alcohols found in hormone and pheromone metabolism . These data suggest that protein functional diversity can expand rapidly under the joint forces of exon shuffling, gene duplication, and natural selection.

J Environ Sci Health A Tox Hazard Subst Environ Eng, 2004, 39(11-12), 2973 - 82
Microbial recovery of copper from printed circuit boards of waste computer by Acidithiobacillus ferrooxidans; Choi MS et al.; The bioleaching of copper contained in the printed circuit boards (PCB) of waste computers by A . ferrooxidans was studied . The Fe oxidation rates by A . ferrooxidans in the 9K medium supplemented with the leachate of PCB (0.15-0.13 g L(-1) d(-1)) were similar to that in the 9K medium without the leachate (0.15 g L(-1) d(-1)) . This finding suggests that the leachate of PCB did not seriously affect the bioleaching process by this bacterium . The amount of copper leached from PCB shreds increased with the addition of ferrous ion and reached up to 5190 mg L(-1) when the initial concentration of Fe2+ ion was 7 g L(-1) . As the microbial leaching progressed, pale brown precipitate was observed to form in the solution . Based on the total amount of copper, both in solution and precipitate, the optimal addition of ferrous ion for the leaching of copper was around 7 g L(-1) . When citric acid was not added, only about 37 wt% of the total leached copper remained dissolved; however, the amount of dissolved copper increased to greater than 80 wt% in the presence of citric acid . This fact indicates that the addition of a complexing agent (citric acid) to the bioleaching solution can raise the solubility of the leached metal ions.

Int J Med Microbiol, 2004 Oct, 294(5), 345 - 54
CpG-DNA as immune response modifier; Dalpke AH et al.; Toll-like receptors (TLRs) have been recognized to play a fundamental role in the recognition of microbial pathogens and the activation of innate immunity . However, it has also turned out that triggering of TLRs with purified compounds or synthetic ligands represents a powerful means to modulate innate as well as adaptive immune responses . Among the substances currently under investigation as immune response modifiers CpG-DNA has gained particular interest . One reason for this is the feasibility to easily synthesize and modify immunostimulatory CpG-containing oligodeoxynucleotides (CpG-ODNs) . This review deals with the structural needs of CpG-ODNs . CpG-ODNs are discussed as being composed of distinct building blocks (e.g . sequence strings, backbone modifications) giving rise to a concept of structural modules within CpG-ODNs.

Nat Immunol, 2004 Dec, 5(12), 1227 - 34 Epub 2004 Dec.
Bone marrow plasmacytoid dendritic cells can differentiate into myeloid dendritic cells upon virus infection; Zuniga EI et al.; Two subsets of dendritic cell (DCs), plasmacytoid (p) and myeloid (m) DCs, have been described in humans and mice . These subsets are known to have divergent roles during an immune response, but their developmental course is unclear . Here we report that virus infection induces bone marrow pDCs to differentiate into mDCs, thereby undergoing profound phenotypic and functional changes including the acquisition of enhanced antigen-presenting capacity and the ability to recognize different microbial structures through Toll-like receptor 4 . The conversion of pDCs into mDCs is also induced by the injection of double-stranded RNA and requires type I interferons . Our results establish a precursor-product developmental relationship between these two DC subsets and highlight unexpected plasticity of bone marrow pDCs.

Indian J Pediatr, 2004 Oct, 71(10), 879 - 82
Diarrhea prevention through food safety education; Sheth M et al.; OBJECTIVE: Food safety education (FSE) was imparted in order to reduce the prevalence of diarrhea in children and improving the knowledge, attitude and practices of mothers regarding safe feeding practices . METHODS: The study was conducted on mothers (n=200) of underprivileged children (6-24 months) through Anganwadi workers (AWW) . The food safety education package incorporated three messages: washing hands with soap and water, avoid feeding leftover food and keep surrounding clean . These were imparted using lectures, slogans, posters, charts, flash cards, role-play . Calendars and leaflets were also handed over to the mothers as education material . After food safety education intervention, the households were studied again as at the baseline, on various aspects . RESULT: There was 52% reduction in the incidence of diarrhea . The environmental sanitation and personal hygiene scores of most of the households and mothers improved . Improvement in the practice of hand-washing and avoidance of feeding leftovers to children was seen . There was reduction in the microbial load in the hand rinse samples of both mothers and children . CONCLUSION: AWW proved to be effective change agents and FSE was successful in bringing about a positive behavior modification in mothers.

Environ Health Perspect, 2004 Nov, 112(15), 1508 - 11
Production of androgens by microbial transformation of progesterone in vitro: a model for androgen production in rivers; Jenkins RL et al.; We have previously documented the presence of progesterone and androstenedione in the water column and bottom sediments of the Fenholloway River, Taylor County, Florida . This river receives paper mill effluent and contains masculinized female mosquitofish . We hypothesized that plant sterols (e.g., ss-sitosterol) derived from the pulping of pine trees are transformed by bacteria into progesterone and subsequently into 17alpha-hydroxyprogesterone, androstenedione, and other androgens . In this study, we demonstrate that these same androgens can be produced in vitro from the bacterium Mycobacterium smegmatis . In a second part to this study, we reextracted and reanalyzed the sediment from the Fenholloway River and verified the presence of androstadienedione, a delta1 steroid with androgen activity.

Trends Immunol, 2004 Dec, 25(12), 677 - 86
The chemokine system in diverse forms of macrophage activation and polarization; Mantovani A et al.; Plasticity and functional polarization are hallmarks of the mononuclear phagocyte system . Here we review emerging key properties of different forms of macrophage activation and polarization (M1, M2a, M2b, M2c), which represent extremes of a continuum . In particular, recent evidence suggests that differential modulation of the chemokine system integrates polarized macrophages in pathways of resistance to, or promotion of, microbial pathogens and tumors, or immunoregulation, tissue repair and remodeling.

Trends Immunol, 2004 Dec, 25(12), 650 - 4
A central role for tissue-resident dendritic cells in innate responses; Foti M et al.; In the past 20 years, dendritic cells (DCs) have been investigated mainly with regard to antigen-specific immune responses and acquired immunity . More recently, new data have indicated a novel and fundamental role for DCs in innate immunity, which is shown by the ability of DCs to induce natural killer (NK) cell activation . In mice, the molecular mechanism by which NK cells are activated by DCs has been revealed, and it consists of the induction of interleukin-2 (IL-2) expression during the early phases of an immune response . Moreover, as indicated by genome-wide transcriptional analysis, microbially activated myeloid DCs produce other chemokines and inflammatory cytokines, such as type I interferons, which in addition to IL-2 are also involved in NK cell activation . This indicates that DCs have a key role in innate immunity . The expression of an innate receptor repertoire on DCs mediates microbial recognition and uptake, and the antigen processing and specific sorting mechanisms lead to very efficient antigen presentation by DCs . Thus, we believe that DCs participate fully in innate responses, forming a bridge between innate and acquired immunity.

J Theor Biol, 2005 Jan 21, 232(2), 285 - 99
Connection between stochastic and deterministic modelling of microbial growth; Kutalik Z et al.; We present in this paper various links between individual and population cell growth . Deterministic models of the lag and subsequent growth of a bacterial population and their connection with stochastic models for the lag and subsequent generation times of individual cells are analysed . We derived the individual lag time distribution inherent in population growth models, which shows that the Baranyi model allows a wide range of shapes for individual lag time distribution . We demonstrate that individual cell lag time distributions cannot be retrieved from population growth data . We also present the results of our investigation on the effect of the mean and variance of the individual lag time and the initial cell number on the mean and variance of the population lag time . These relationships are analysed theoretically, and their consequence for predictive microbiology research is discussed.

Chem Phys Lipids, 2004 Nov, 132(1), 113 - 32
Binding and insertion of alpha-helical anti-microbial peptides in POPC bilayers studied by molecular dynamics simulations; Kandasamy SK et al.; We have performed molecular dynamics simulations of the interactions of two alpha-helical anti-microbial peptides, magainin2 and its synthetic analog of MSI-78, with palmitoyl-oleoyl-phosphatidylcholine (POPC) lipid bilayers . We used various initial positions and orientations of the peptide with respect to the lipid bilayer, including a surface-bound state parallel to the interface, a trans-membrane state, and a partially inserted state . Our 20 ns long simulations show that both magainin2 and MSI-78 are most stable in the lipid environment, with the peptide destabilized to different extents in both aqueous and lipid/water interfacial environments . We found that there are strong specific interactions between the lysine residues of the peptides and the lipid head-group regions . MSI-78, owing to its large number of lysines, shows better binding characteristics and overall stability when compared to magainin2 . We also find that both peptides destabilize the bilayer environment, as observed by the increase in lipid tail disorder and the induction of local curvature on the lipid head-groups by the peptides . From all the simulations, we conclude that the hydrogen bonding interactions between the lysines of the peptides and the oxygens of the polar lipid head-groups are the strongest and determine the overall peptide binding characteristics to the lipids.

Structure (Camb), 2004 Nov, 12(11), 2015 - 24
Structures of Selenomonas ruminantium phytase in complex with persulfated phytate: DSP phytase fold and mechanism for sequential substrate hydrolysis; Chu HM et al.; Various inositide phosphatases participate in the regulation of inositol polyphosphate signaling molecules . Plant phytases are phosphatases that hydrolyze phytate to less-phosphorylated myo-inositol derivatives and phosphate . The phytase from Selenomonas ruminantium shares no sequence homology with other microbial phytases . Its crystal structure revealed a phytase fold of the dual-specificity phosphatase type . The active site is located near a conserved cysteine-containing (Cys241) P loop . We also solved two other crystal forms in which an inhibitor, myo-inositol hexasulfate, is cocrystallized with the enzyme . In the "standby" and the "inhibited" crystal forms, the inhibitor is bound, respectively, in a pocket slightly away from Cys241 and at the substrate binding site where the phosphate group to be hydrolyzed is held close to the -SH group of Cys241 . Our structural and mutagenesis studies allow us to visualize the way in which the P loop-containing phytase attracts and hydrolyzes the substrate (phytate) sequentially.

Folia Microbiol (Praha), 2004, 49(4), 339 - 52
Natural microbial UV radiation filters--mycosporine-like amino acids; Rezanka T et al.; Ozone depletion by anthropogenic gases has increased the atmospheric transmission of solar ultraviolet-B radiation (UV-B, 280-315 nm) . There is a logical link between the natural defenses of terrestrial and marine organisms against UV radiation and the prevention of UV-induced damage to human skin . UV light degrades organic molecules such as proteins and nucleic acids, giving rise to structural changes that directly affect their biological function . These compounds offer the potential for development of novel UV blockers for human use . The biological role of mycosporine-like amino acids (MAAs) and scytonemin as a defense against solar radiation in organisms, together with their structure, synthesis, distribution, regulation and effectiveness, are reviewed in this article . This review points to the role of MAAs as a natural defense against UV radiation.

J Environ Biol, 2004 Apr, 25(2), 201 - 8
Pesticide effects on microbial population and soil enzyme activities during vermicomposting of agricultural waste; Kalam A et al.; The effects of different pesticides (propiconazole, profenofos, pretilachlor) on vermicomposting were evaluated for the adverse effects on soil enzyme activities (dehydrogenase, phosphatase, urease) and total microbial counts (TMC) . There were remarkable increase in enzyme activities and TMC in presence of earthworm compared to control (earthworm absent) . In comparison to control, the activities of phosphatase, dehydrogenase and urease in presence of vermicompost increased upto 30, 128 and 31.3% respectively; whereas increase of TMC was 71.9% . But, in presence of each of the pesticide, said activities decreased . Maximum inhibition of soil phosphatase activity (46.6%) was observed in presence of propiconazole (100 mg kg(-1)) after 120 days . Profenofos affected the soil dehydrogenase activity in the tune of 47% at 1000 mg kg(-1) concentration after 80 days and thereafter, the extent of toxicity decreased little . Soil urease activity was affected markedly in presence of profenofos and was 62% at 1000 mg kg(-1) level after 80 days . TMC also declined in presence of profenofos and pretilachlor . Increase in TMC was about 71.9% compared to control and the inhibition was more or less 60% when profenofos (1000 mg kg(-1)) was present even after 120 days of treatment.

Nat Biotechnol, 2004 Nov, 22(11), 1415 - 22
Plant cell cultures for the production of recombinant proteins; Hellwig S et al.; The use of whole plants for the synthesis of recombinant proteins has received a great deal of attention recently because of advantages in economy, scalability and safety compared with traditional microbial and mammalian production systems . However, production systems that use whole plants lack several of the intrinsic benefits of cultured cells, including the precise control over growth conditions, batch-to-batch product consistency, a high level of containment and the ability to produce recombinant proteins in compliance with good manufacturing practice . Plant cell cultures combine the merits of whole-plant systems with those of microbial and animal cell cultures, and already have an established track record for the production of valuable therapeutic secondary metabolites . Although no recombinant proteins have yet been produced commercially using plant cell cultures, there have been many proof-of-principle studies and several companies are investigating the commercial feasibility of such production systems.

J Clin Microbiol, 2004 Nov, 42(11), 5170 - 5
Analysis of vaginal acetic acid in patients undergoing treatment for bacterial vaginosis; Chaudry AN et al.; A "gold standard" method for the diagnosis of bacterial vaginosis (BV) is lacking . The clinical criteria described by the Amsel technique are subjective and difficult to quantify . Alternatively, the reading of Gram-stained vaginal smears by scoring techniques such as those that use the Nugent or Hay-Ison scoring systems is again subjective, requires expert personnel to perform the reading, and is infrequently used clinically . Recently, a new diagnostic device, the Osmetech Microbial Analyzer--Bacterial Vaginosis (OMA-BV), which determines a patient's BV status on the basis of measurement of the amount of acetic acid present in a vaginal swab specimen, was approved by the Food and Drug Administration . The present study uses the conducting polymer gas-sensing technology of OMA-BV to measure the concentration of acetic acid in the headspace above vaginal swab specimens from patients undergoing treatment for BV with metronidazole . In 97.8% of the cases the level of acetic acid detected fell sharply during the treatment period, crossing from above to below the diagnostic threshold of 900 ppm . The diagnosis obtained on the basis of the level of vaginal acetic acid was compared with the diagnoses obtained by use of the Amsel criteria and the Nugent scoring system both at the time of initial entry into the study and at the repeat samplings on days 7 and 14 . The results obtained with OMA-BV showed overall agreements compared with the results of the Amsel and Nugent tests of 98 and 94%, respectively, for the 34 patients monitored through the treatment process . This provides further evidence that the measurement of vaginal acetic acid by headspace analysis with conducting polymer sensors is a valid alternative to present tests for the diagnosis of BV.

Appl Environ Microbiol, 2004 Nov, 70(11), 6569 - 79
Diversity and antagonistic potential of bacteria associated with bryophytes from nutrient-poor habitats of the Baltic Sea Coast; Opelt K et al.; Very little is known about the interaction of bryophytes with bacteria . Therefore, we analyzed bacteria associated with three bryophyte species, Tortula ruralis, Aulacomnium palustre, and Sphagnum rubellum, which represent typical moss species of three nutrient-poor plant communities at the southern Baltic Sea coast in Germany . By use of two cultivation-independent techniques, denaturing gradient gel electrophoresis and single-strand conformation polymorphism analysis of the 16S ribosomal DNA, a high degree of moss specificity was found for associated bacterial communities . This specificity could be further evidenced by a cultivation-dependent approach for the following parameters: (i) plate counts of bacteria on R2A medium, (ii) proportion of antagonistic isolates, (iii) antagonistic activity as well as spectrum against pathogens, and (iv) diversity and richness of antagonistic isolates . The proportion of isolates with antagonistic activity against the pathogenic model fungus Verticillium dahliae was highest for S . rubellum (31%), followed by A . palustre (17%) and T . ruralis (5%) . A high percentage (99%) of moss-associated antagonistic bacteria produced antifungal compounds . The high recovery of antagonistic isolates strongly suggests that bryophytes represent an ecological niche which harbors a diverse and hitherto largely uncharacterized microbial population with yet unknown and untapped potential biotechnological applications, e.g., for biological control of plant pathogens.

Appl Environ Microbiol, 2004 Nov, 70(11), 6428 - 34
Redox transformations of arsenic oxyanions in periphyton communities; Kulp TR et al.; Periphyton (Cladophora sp.) samples from a suburban stream lacking detectable dissolved As were able to reduce added As(V) to As(III) when incubated under anoxic conditions and, conversely, oxidized added As(III) to As(V) with aerobic incubation . Both types of activity were abolished in autoclaved controls, thereby demonstrating its biological nature . The reduction of As(V) was inhibited by chloramphenicol, indicating that it required the synthesis of new protein . Nitrate also inhibited As(V) reduction, primarily because it served as a preferred electron acceptor to which the periphyton community was already adapted . However, part of the inhibition was also caused by microbial reoxidation of As(III) linked to nitrate . Addition of {14C}glucose to anoxic samples resulted in the production of 14CO2, suggesting that the observed As(V) reduction was a respiratory process coupled to the oxidation of organic matter . The population density of As(V)-reducing bacteria within the periphyton increased with time and with the amount of As(V) added, reaching values as high as approximately 10(6) cells ml(-1) at the end of the incubation . This indicated that dissimilatory As(V) reduction in these populations was linked to growth . However, As(V)-respiring bacteria were found to be present, albeit at lower numbers (approximately 10(2) ml(-1)), in freshly sampled periphyton . These results demonstrate the presence of a bacterial population within the periphyton communities that is capable of two key arsenic redox transformations that were previously studied in As-contaminated environments, which suggests that these processes are widely distributed in nature . This assumption was reinforced by experiments with estuarine samples of Cladophora sericea in which we detected a similar capacity for anaerobic As(V) reduction and aerobic As(III) oxidation.

Appl Environ Microbiol, 2004 Nov, 70(11), 6407 - 13
Effect of microbial inoculants on the indigenous actinobacterial endophyte population in the roots of wheat as determined by terminal restriction fragment length polymorphism; Conn VM et al.; The effect of single actinobacterial endophyte seed inoculants and a mixed microbial soil inoculant on the indigenous endophytic actinobacterial population in wheat roots was investigated by using the molecular technique terminal restriction fragment length polymorphism (T-RFLP) . Wheat was cultivated either from seeds coated with the spores of single pure actinobacterial endophytes of Microbispora sp . strain EN2, Streptomyces sp . strain EN27, and Nocardioides albus EN46 or from untreated seeds sown in soil with and without a commercial mixed microbial soil inoculant . The endophytic actinobacterial population within the roots of 6-week-old wheat plants was assessed by T-RFLP . Colonization of the wheat roots by the inoculated actinobacterial endophytes was detected by T-RFLP, as were 28 to 42 indigenous actinobacterial genera present in the inoculated and uninoculated plants . The presence of the commercial mixed inoculant in the soil reduced the endophytic actinobacterial diversity from 40 genera to 21 genera and reduced the detectable root colonization by approximately half . The results indicate that the addition of a nonadapted microbial inoculum to the soil disrupted the natural actinobacterial endophyte population, reducing diversity and colonization levels . This was in contrast to the addition of a single actinobacterial endophyte to the wheat plant, where the increase in colonization level could be confirmed even though the indigenous endophyte population was not adversely affected.

Appetite, 2004 Dec, 43(3), 309 - 13
Consumer perceptions of food safety risk, control and responsibility; Redmond EC et al.; Consumer perceptions of risk, control and responsibility related to microbial food safety in domestic food preparation have been investigated . Results showed that consumers demonstrated judgements of 'optimistic-bias' and the 'illusion of control', as well as notions of perceived invulnerability to food poisoning from self-prepared foods . Statistical associations between perceptions of personal risk, control and responsibility and risk and control attributed to 'other people' have been identified . Findings may have negative implications for the effectiveness of consumer food safety education initiatives . It is suggested that consideration of such judgements and associations during the development of future consumer food safety risk communication strategies may increase their effectiveness.

Australas J Dermatol, 2004 Nov, 45(4), 199 - 205; quiz 206-7
Malassezia yeasts in the pathogenesis of atopic dermatitis; Aspres N et al.; Atopic dermatitis is a common skin condition, the aetiology of which is multifactorial, involving genetic, immunological and environmental factors . In recent years, it has been suggested that various microbial organisms may also be involved in the pathogenesis of the disease . Yeasts belonging to the Malassezia genus have received particular attention . These yeasts, known to be a part of the normal skin flora, have been shown to be capable of inducing immunoglobulin (Ig)E-mediated and T-cell mediated immune responses postulated to contribute to chronic inflammation in the skin, particularly in the head and neck region, where colonization with Malassezia is the greatest . Considerable evidence now exists to support this idea, raising the possibility that specific antifungal therapy may be a useful treatment measure in some atopic patients who have a head and neck pattern of dermatitis.

Cell Biol Educ, 2004 Fall, 3(3), 155 - 61
PowerPoint in the classroom; Risk Analysis of Antibiotic Resistance et al.; Joint FAO/WHO Food Standards Programme, FAO, Viale delle Terme di Caracalla, Rome, Italy . selma.doyran@fao.org

The FAO/WHO Codex Alimentarius Commission develops recommendations concerning risk management related to regulatory issues, on the basis of scientific advice provided by FAO and WHO . The complex question of anti-microbial resistance that requires a multidisciplinary approach, has been considered from different point of views in the framework of Codex, especially food hygiene and veterinary drugs and to a lesser extent, plant protection . Antibiotic resistance was considered from the perspective of the food chain and a risk profile was developed . Risk management options were also considered and may be grouped in two main categories: prevention of contamination through the food chain; and prudent use of anti-microbials . It is recognized that further scientific advice is necessary before developing specific risk management options in the framework of Codex . It is expected that the results of the FAO/OIE/WHO Expert Consultations on anti-microbial resistance will provide the necessary guidance for further action.

J Vet Med B Infect Dis Vet Public Health, 2004 Oct-Nov, 51(8-9), 400 - 2
Microbial Threat--The Copenhagen Recommendations initiative of the EU; Frimodt-Moller N; The Microbial Threat was the name of an invitational EU conference held in Copenhagen in 1998, which for the first time made antibiotic resistance an official EU issue . The initiative resulted in a set of conclusions, which together with a summary of the conference was published as The Copenhagen Recommendations . This was soon followed by an EU Council Recommendation in 2001 exhorting the member states to follow and enact the recommendations from the Copenhagen meeting . Funding from EU research funds was later provided for projects monitoring resistance as well as antibiotic use, both of which have been lacking in most EU countries until now . The initiative is a good example of a major health issue being lifted by a concerted and official EU action.

J Vet Med B Infect Dis Vet Public Health, 2004 Oct-Nov, 51(8-9), 357 - 60
Towards the rational use of antibiotics: results of the first International Symposium on the Risk Analysis of Antibiotic Resistance; Helmuth R et al.; The results of four expert working groups on the risk analysis process on anti-microbial resistance are described . They focus on the procedure recommended by the FAO/WHO CODEX Alimentarius Commission in 1999 including hazard identification, hazard characterization, exposure assessment and risk characterization . In all four areas, the current scientific knowledge was evaluated and recommendations for risk management options were given . They will form the basis for mitigating the health risk caused by resistant micro-organisms.

Anal Sci, 2004 Oct, 20(10), 1453 - 7
Structural features of humic acid of the coastal sediment in Ariake Sea tidelands: use of humic acid as an environmental indicator for river basins and coastal regions; Yamauchi N et al.; The structural features of humic acid (HA) at the sediment surface of the tideland at the Hayatsuegawa-river mouth at the Ariake Sea were investigated for the utilization of HA toward an environmental indicator of the features of the river basin and coastal region . 1H NMR analysis revealed a high-content hydrocarbon residue with a similar type of terrigenous HA . Direct and methylation-pyrolysis-GC analysis suggested the incorporation of long-chain carboxylate in HA in the tidelands . The incorporation of branched-chain carboxylate residues in HA is the result of the microbial decomposition of detritus; these residues could be one of the characteristic structural features of HA in this area, which is rich in biodiversity and microbial activity . Because the structural features of coastal zone HA appear to reveal the characteristics and activities of the biological environment, these findings suggest the possibility of becoming an indicator of the detailed analysis of the structural features of coastal zone HA.

Biodegradation, 2004 Oct, 15(5), 315 - 25
Effect of iron bioavailability on dissolved hydrogen concentrations during microbial iron reduction; Komlos J et al.; Dissolved hydrogen (H2) concentrations have been shown to correlate with specific terminal electron accepting processes (TEAPs) in aquifers . The research presented herein examined the effect of iron bioavailability on H2 concentrations during iron reduction in flow-through column experiments filled with soil obtained from the uncontaminated background area of the Field Research Center (FRC), Oak Ridge, TN and amended with acetate as the electron donor . The first column experiment measured H2 concentrations over 500 days of column operation that fluctuated within a substantial range around an average of 3.9 nM . Iron reduction was determined to be the dominant electron accepting process . AQDS (9,10-anthraquinone-2,6-disulfonic acid) was then used to determine if H2 concentrations during iron reduction were related to iron bioavailability . For this purpose, a 100-day flow-through column experiment was conducted that compared the effect of AQDS on iron reduction and subsequent H2 concentrations using two columns in parallel . Both columns were packed with FRC soil and inoculated with Geobacter sulfurreducens but only one was supplied with AQDS . The addition of AQDS increased the rate of iron reduction in the flow-through column and slightly decreased the steady-state H2 concentrations from an average of 4.0 nM for the column without AQDS to 2.0 nM for the column with AQDS . The results of this study therefore show that H2 can be used as an indicator to monitor rate and bioavailability changes during microbial iron reduction.

Presse Med, 2004 Oct 9, 33(17), 1164 - 8
Prognostic factors of hematological recovery in life-threatening nonchemotherapy drug-induced agranulocytosis . A study of 91 patients from a single center; Maloisel F et al.; OBJECTIVES: We studied clinical factors that may influence the duration of hematological recovery to reach neutrophil counts and thus, indirectly, the prognosis in patients with life-threatening drug-induced agranulocytosis (DIA) . METHODS: Using univariate and multivariate analyses with Cox's proportional hazard models, we determined the prognostic factors for hematological recovery, defined as neutrophil counts>0.5 and>1.5.10(9)/L, in 91 patients with established life-threatening DIA . RESULTS: Multivariable analysis showed that neutrophil count<0.1.10(9)/L (at diagnosis) and infection profile: severe infections or septic shock, adversely influenced the neutrophil recovery (for the two neutrophil levels) . Hematopoietic growth factors were significantly associated with rapid hematological recovery (for the two neutrophil levels) . Documented microbial infections and antiplatelet DIA were also associated with rapid hematological recovery (for a neutrophil count>1.5.10(9)/L) . CONCLUSION: Our findings demonstrate that in life-threatening DIA, hematological recovery is mainly dependent of the neutrophil level, the type of infections and the utilization of hematopoietic growth factors.

J Biol Chem, 2005 Jan 14, 280(2), 1678 - 87 Epub 2004 Nov 02.
Stimulus-specific Induction of a Novel Nuclear Factor-{kappa}B Regulator, I{kappa}B-{zeta}, via Toll/Interleukin-1 Receptor Is Mediated by mRNA Stabilization; Yamazaki S et al.; We have recently identified an inducible nuclear factor-kappaB (NF-kappaB) regulator, IkappaB-zeta, which is induced by microbial ligands for Toll-like receptors such as lipopolysaccharide and the proinflammatory cytokine interleukin (IL)-1beta but not by tumor necrosis factor (TNF)-alpha . In the present study, we examined mechanisms for stimulus-specific induction of IkappaB-zeta . The analysis of the IkappaB-zeta promoter revealed an essential role for an NF-kappaB binding sequence in transcriptional activation . The activation, however, did not account for the Toll-like receptor/IL-1 receptor-specific induction of IkappaB-zeta, because the promoter analysis and nuclear run-on analysis indicated that its transcription was similarly induced by TNF-alpha . To examine post-transcriptional regulation, we analyzed the decay of IkappaB-zeta mRNA, and we found that it was specifically stabilized by lipopolysaccharide or IL-1beta but not by TNF-alpha . Furthermore, we found that costimulation with TNF-alpha and another proinflammatory cytokine, IL-17, elicited the IkappaB-zeta induction . Stimulation with IL-17 alone did not induce IkappaB-zeta but stabilized its mRNA . Therefore, IkappaB-zeta induction requires both NF-kappaB activation and stimulus-specific stabilization of its mRNA . Because IkappaB-zeta is essential for expression of a subset of NF-kappaB target genes, the stimulus-specific induction of IkappaB-zeta may be of great significance in regulation of inflammatory reactions.

Gastroenterology, 2004 Nov, 127(5), 1474 - 87
Probiotics inhibit nuclear factor-kappaB and induce heat shock proteins in colonic epithelial cells through proteasome inhibition; Petrof EO et al.; BACKGROUND AND AIMS: The extent and severity of mucosal injury in inflammatory bowel diseases are determined by the disequilibrium between 2 opposing processes: reparative and cytoprotective mechanisms vs . inflammation-induced injury . Probiotics may provide clinical benefit by ameliorating colitis; however, their mechanisms of action remain largely unknown . Our objective was to investigate microbial-epithelial interactions that could explain the beneficial therapeutic effects of probiotics . METHODS: The effect of VSL#3-conditioned media on the nuclear factor-kappaB pathway in young adult mouse colonic epithelial cells was assessed by using monocyte chemoattractant protein-1 enzyme-linked immunosorbent assays; IkappaBalpha, IkappaBbeta, and p105 immunoblot analysis; and nuclear factor-kappaB luciferase reporter gene and proteasome assays . Effects on heat shock proteins were determined by electrophoretic mobility shift assay and immunoblot for heat shock proteins 25 and 72 in young adult mouse colonic cells . Cytoprotection against oxidant injury was determined by chromium 51 release and filamentous and globular actin assays . RESULTS: VSL#3 produces soluble factors that inhibit the chymotrypsin-like activity of the proteasome in gut epithelial cells . Proteasome inhibition is an early event that begins almost immediately after exposure of the epithelial cells to the probiotic-conditioned media . In addition, these bacteria inhibit the proinflammatory nuclear factor-kappaB pathway through a mechanism different from the type III secretory mechanisms described for other nonpathogenic enteric flora . They also induce the expression of cytoprotective heat shock proteins in intestinal epithelial cells . CONCLUSIONS: The resulting inhibition of nuclear factor-kappaB and increased expression of heat shock proteins may account for the anti-inflammatory and cytoprotective effects reported for probiotics and may be a novel mechanism of microbial-epithelial interaction . These effects seem to be mediated through the common unifying mechanism of proteasome inhibition.

Genome Res, 2004 Nov, 14(11), 2367 - 76
OptStrain: a computational framework for redesign of microbial production systems; Pharkya P et al.; This paper introduces the hierarchical computational framework OptStrain aimed at guiding pathway modifications, through reaction additions and deletions, of microbial networks for the overproduction of targeted compounds . These compounds may range from electrons or hydrogen in biofuel cell and environmental applications to complex drug precursor molecules . A comprehensive database of biotransformations, referred to as the Universal database (with >5700 reactions), is compiled and regularly updated by downloading and curating reactions from multiple biopathway database sources . Combinatorial optimization is then used to elucidate the set(s) of non-native functionalities, extracted from this Universal database, to add to the examined production host for enabling the desired product formation . Subsequently, competing functionalities that divert flux away from the targeted product are identified and removed to ensure higher product yields coupled with growth . This work represents an advancement over earlier efforts by establishing an integrated computational framework capable of constructing stoichiometrically balanced pathways, imposing maximum product yield requirements, pinpointing the optimal substrate(s), and evaluating different microbial hosts . The range and utility of OptStrain are demonstrated by addressing two very different product molecules . The hydrogen case study pinpoints reaction elimination strategies for improving hydrogen yields using two different substrates for three separate production hosts . In contrast, the vanillin study primarily showcases which non-native pathways need to be added into Escherichia coli . In summary, OptStrain provides a useful tool to aid microbial strain design and, more importantly, it establishes an integrated framework to accommodate future modeling developments.

Tree Physiol, 2005 Jan, 25(1), 57 - 66
Forest thinning and soil respiration in a ponderosa pine plantation in the Sierra Nevada; Tang J et al.; Soil respiration is controlled by soil temperature, soil water, fine roots, microbial activity, and soil physical and chemical properties . Forest thinning changes soil temperature, soil water content, and root density and activity, and thus changes soil respiration . We measured soil respiration monthly and soil temperature and volumetric soil water continuously in a young ponderosa pine (Pinus ponderosa Dougl . ex P . Laws . & C . Laws.) plantation in the Sierra Nevada Mountains in California from June 1998 to May 2000 (before a thinning that removed 30% of the biomass), and from May to December 2001 (after thinning) . Thinning increased the spatial homogeneity of soil temperature and respiration . We conducted a multivariate analysis with two independent variables of soil temperature and water and a categorical variable representing the thinning event to simulate soil respiration and assess the effect of thinning . Thinning did not change the sensitivity of soil respiration to temperature or to water, but decreased total soil respiration by 13% at a given temperature and water content . This decrease in soil respiration was likely associated with the decrease in root density after thinning . With a model driven by continuous soil temperature and water time series, we estimated that total soil respiration was 948, 949 and 831 g C m(-2) year(-1) in the years 1999, 2000 and 2001, respectively . Although thinning reduced soil respiration at a given temperature and water content, because of natural climate variability and the thinning effect on soil temperature and water, actual cumulative soil respiration showed no clear trend following thinning . We conclude that the effect of forest thinning on soil respiration is the combined result of a decrease in root respiration, an increase in soil organic matter, and changes in soil temperature and water due to both thinning and interannual climate variability.

Environ Pollut, 2005 Feb, 133(3), 455 - 65
The effects of perennial ryegrass and alfalfa on microbial abundance and diversity in petroleum contaminated soil; Kirk JL et al.; Enhanced rhizosphere degradation uses plants to stimulate the rhizosphere microbial community to degrade organic contaminants . We measured changes in microbial communities caused by the addition of two species of plants in a soil contaminated with 31,000 ppm of total petroleum hydrocarbons . Perennial ryegrass and/or alfalfa increased the number of rhizosphere bacteria in the hydrocarbon-contaminated soil . These plants also increased the number of bacteria capable of petroleum degradation as estimated by the most probable number (MPN) method . Eco-Biolog plates did not detect changes in metabolic diversity between bulk and rhizosphere samples but denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified partial 16S rDNA sequences indicated a shift in the bacterial community in the rhizosphere samples . Dice coefficient matrices derived from DGGE profiles showed similarities between the rhizospheres of alfalfa and perennial ryegrass/alfalfa mixture in the contaminated soil at week seven . Perennial ryegrass and perennial ryegrass/alfalfa mixture caused the greatest change in the rhizosphere bacterial community as determined by DGGE analysis . We concluded that plants altered the microbial population; these changes were plant-specific and could contribute to degradation of petroleum hydrocarbons in contaminated soil.

Adv Drug Deliv Rev, 2005 Jan 2, 57(1), 123 - 34
Use of fluorescence imaging to investigate the structure and function of intestinal M cells; Buda A et al.; Fluorescence imaging technology can be applied to many aspects of cell biology ranging from the analysis of specific markers in cells and tissues to the biological actions and distribution of fluorescent proteins or particles in living cells . In this review, we examine the role of fluorescence imaging, in conjunction with other microscopical techniques, to study sites of uptake of material across the gastrointestinal epithelium . We will focus primarily on intestinal M cells, specialised antigen-sampling cells in the epithelium of the gut-associated lymphoid tissue (GALT), including Peyer's patches . In addition to their importance as sites for uptake of inert material, and hence their potential as a route of delivery of vaccines, etc., M cells are also a major site of infection by a range of microbial pathogens . The application of new fluorescence imaging technologies has expanded our knowledge on the structure, development and function of these fascinating cells.

Virology, 2004 Nov 24, 329(2), 412 - 24
Improved isolation of undersampled bacteriophages: finding of distant terminase genes; Serwer P et al.; Isolation and characterization of new environmental bacteriophages are performed for (1) analyzing microbial evolution and ecology and (2) delivering biological therapy . The sampling of environmental bacteriophages appears, however, to be limited by the procedure (usually liquid enrichment culture) used to propagate them . An alternative, less competitive procedure is developed here for the purpose of isolating new bacteriophages . This procedure involves extraction directly into and then propagation in a dilute agarose gel . Adaptations of this procedure are used to avoid repeated isolation of the same bacteriophage . Some newly isolated bacteriophages grow so poorly that they appear inaccessible to liquid enrichment culture . Four comparatively high titer bacteriophages were isolated and characterized by a genomic sequence survey . Some had genomes with extremely distant relationships to those of other bacteriophages, based on a tree built from the large terminase genes . These methods find novel genomes by rapidly isolating and screening diverse bacteriophages.

Virology, 2004 Nov 24, 329(2), 251 - 60
Differential roles of CCL2 and CCR2 in host defense to coronavirus infection; Held KS et al.; The CC chemokine ligand 2 (CCL2, monocyte chemoattractant protein-1) is important in coordinating the immune response following microbial infection by regulating T cell polarization as well as leukocyte migration and accumulation within infected tissues . The present study examines the consequences of mouse hepatitis virus (MHV) infection in mice lacking CCL2 (CCL2(-/-)) in order to determine if signaling by this chemokine is relevant in host defense . Intracerebral infection of CCL2(-/-) mice with MHV did not result in increased morbidity or mortality as compared to either wild type or CCR2(-/-) mice and CCL2(-/-) mice cleared replicating virus from the brain . In contrast, CCR2(-/-) mice displayed an impaired ability to clear virus from the brain that was accompanied by a reduction in the numbers of antigen-specific T cells as compared to both CCL2(-/-) and wild-type mice . The paucity in T cell accumulation within the central nervous system (CNS) of MHV-infected CCR2(-/-) mice was not the result of either a deficiency in antigen-presenting cell (APC) accumulation within draining cervical lymph nodes (CLN) or the generation of virus-specific T cells within this compartment . A similar reduction in macrophage infiltration into the CNS was observed in both CCL2(-/-) and CCR2(-/-) mice when compared to wild-type mice, indicating that both CCL2 and CC chemokine receptor 2 (CCR2) contribute to macrophage migration and accumulation within the CNS following MHV infection . Together, these data demonstrate that CCR2, but not CCL2, is important in host defense following viral infection of the CNS, and CCR2 ligand(s), other than CCL2, participates in generating a protective response.

J Rheumatol, 2004 Nov, 31(11), 2300 - 4
Common aspects of human and primate seronegative arthritis; Hyrich KL et al.; A 27-year-old female lowland gorilla developed an asymmetric oligoarthritis 3 months post-partum.There was no evidence of an antecedent gastrointestinal or genitourinary infection . Serum was negative for rheumatoid factor and antinuclear antibody . Synovial fluid revealed 2000 white blood cells with negative cultures and polarized microscopy . Studies on synoviocytes were the following: (1) FACS analysis revealed surface expression of a B27-like epitope of the cells . (2) Analysis of intracellular clearance kinetics of arthritogenic organisms showed peak intracellular colony-forming units at 48 hours after bacterial invasion, and clearance by 13 days post-invasion . (3) Interferon-y (0.1-10.0 ng/ml)accelerated intracellular microbicidal pathways in a dose-dependent fashion . These findings closely parallel those seen in human synoviocytes of patients with spondyloarthropathy . Primate and human seronegative arthritis share clinical and immunologic features, as well as aspects of host:pathogen defense mechanisms . The interplay of genetic and microbial factors underlying this arthritis appears to be conserved across these species boundaries.

J Ind Microbiol Biotechnol, 2004 Nov, 31(10), 482 - 8 Epub 2004 Nov.
Rapid identification of microbial VOCs from tobacco molds using closed-loop stripping and gas chromatography/time-of-flight mass spectrometry; Meruva NK et al.; Several microbial volatile organic compounds (MVOCs) that can serve as potential chemical markers for microbial contamination in tobacco have been identified . Four different fungal species, Aspergillus niger (AN), A . ornatus (AO), Pencillium chrysogenum (PC) and Rhizopus stolonifer (RS), commonly reported in moldy tobacco were cultured and screened for MVOCs . Because the MVOCs emitted by a microbial species are substrate specific, the fungal strains were separately grown on potato dextrose agar (PDA) and tobacco products . MVOCs from the mold cultures grown on PDA and tobacco products were extracted using closed-loop stripping analysis (CLSA) and identified by gas chromatography/time-of-flight mass spectrometry (GC/TOF-MS) . Some of the prominent tobacco mold markers identified by this method include: 1-octen-3-ol; 2-octen-1-ol; 2-methyl-1-butanol; 3-methyl-1-butanol; 1-octene and 2-pentanone . In particular, 1-octen-3-ol was detected in all the mold cultures and moldy tobacco samples analyzed . Olfactory evaluation of 1-octen-3-ol indicated a characteristic musty odor and the odor threshold was determined to be approximately 200 ng/ml . The limits of detection for 1-octen-3-ol using GC/TOF-MS and GC/mass selective detector (MSD) in the full-scan mode and selected ion monitoring (SIM) mode were investigated . The CLSA-GC/TOF-MS demonstrates a fast, sensitive and semi-quantitative analytical technique for screening tobacco materials for the presence of mold via chemical markers of microbial contamination.

Nat Rev Immunol, 2004 Nov, 4(11), 856 - 67
Strategies to enhance T-cell reconstitution in immunocompromised patients; van den Brink MR et al.; Immune deficiency, together with its associated risks such as infections, is becoming an increasingly important clinical problem owing to the ageing of the general population and the increasing number of patients with HIV/AIDS, malignancies (especially those treated with intensive chemotherapy or radiotherapy) or transplants (of either solid organs or haematopoietic stem cells) . Of all immune cells, T cells are the most often affected, leading to a prolonged deficiency of T cells, which has important clinical consequences . Accordingly, strategies to improve the recovery and function of T cells, as we discuss here, should have a direct impact on reducing the morbidity and mortality of many patients and should increase the efficacy of therapeutic and prophylactic vaccinations against microbial pathogens or tumours.

J Periodontol, 2004 Sep, 75(9), 1203 - 8
Gingival crevicular fluid levels of interleukin-1beta and glycemic control in patients with chronic periodontitis and type 2 diabetes; Engebretson SP et al.; BACKGROUND: Patients with diabetes have increased incidence and severity of periodontal disease not accounted for by differences in the subgingival microbial infection . Poor glycemic control has been consistently associated with periodontal disease severity . Also, recent evidence suggests that hyperglycemia may induce inflammatory cytokine production . Few studies, however, have examined local biochemical measures of periodontal inflammation in patients with type 2 diabetes . The aim of this study was to determine whether glycemic control was related to gingival crevicular fluid (GCF) levels of interleukin-1beta (IL-1beta) . METHODs: GCF samples were collected from 45 patients with type 2 diabetes and untreated chronic periodontitis . Plaque index (PI), bleeding on probing (BOP), probing depth (PD), and attachment level (AL) were recorded at six sites per tooth . IL-1beta levels were determined from individual GCF samples by enzyme-linked immunoabsorbent assay (ELISA) . Individual site and mean patient values were calculated . Glycated hemoglobin (HbA1c) levels were measured from anticoagulated whole blood using an automated affinity chromatography system . Serum glucose was also determined . RESULTS: Clinical periodontal measures (PD, AL, BOP) and measures of glycemic control (HbA1c, random glucose) were significantly correlated with GCF IL-1beta . Patients with greater than 8% HbA1c had significantly higher mean GCF IL-1beta levels than patients with less than 8% HbA1c . In a multivariate model adjusting for age, gender, PD, AL, BOP, and PI, HbA1c and random glucose were independent predictors of high GCF IL-1beta . CONCLUSIONS: Poor glycemic control is associated with elevated GCF IL-1beta . These data are consistent with the hypothesis that hyperglycemia contributes to an heightened inflammatory response, and suggests a mechanism to account for the association between poor glycemic control and periodontal destruction.

Semin Ophthalmol, 2003 Sep, 18(3), 109 - 15
Current management strategies of acute post-operative endophthalmitis; Das T et al.; The current management of acute post-operative endophthalmitis relies on microbial confirmation, intravitreal antibiotics, and vitrectomy . The endophthalmitis vitrectomy study (EVS) demonstrated that systemic antibiotics are not necessary in post-cataract/secondary intraocular lens (IOL) acute bacterial endophthalmitis, and that vitrectomy could be required only for eyes with presenting vision of light perception . Since the publication of the EVS newer tests are available for microbial confirmation, and many new and effective antibiotics are discovered for systemic therapy . We have studied large numbers of patients with endophthalmitis over several years . Perhaps our experience represents the endophthalmitis status in the developing countries . The article is a synthesis of recent publications in the world literature . Finally the authors have suggested a management strategy for acute post-cataract acute bacterial endophthalmitis.

Acta Odontol Scand, 2004 Aug, 62(4), 193 - 8
Consistency of DIAGNOdent instruments for clinical assessment of fissure caries; Astvaldsdottir A et al.; DIAGNOdent (KaVo, Biberach, Germany) has shown high diagnostic accuracy and intra-operator agreement for caries detection, both in vitro and in vivo . The aims of this study were to compare DIAGNOdent with visual examination (VI) and bitewing radiographs (BW) for clinical assessment of occlusal fissures, and to evaluate inter-device consistency of clinical recording using four different DIAGNOdent instruments; secondly, to correlate DIAGNOdent readings with microbial culture of the measured site . The subjects were young adults, the material comprising 34 occlusal fissures, scheduled for restorative treatment at the Dental School Clinic of the University of Iceland . Two examiners conducted visual and radiographic assessments . One examiner measured each site with four DIAGNOdent instruments in random order . The fissure was then opened and lesion depth was classified on a 4-point scale . Bacterial samples were taken from the fissure before and after opening . Intra-operator agreement was high (r = 0.85-0.98) . Inter-device correlation for the four DIAGNOdent instruments was significant in all cases (r = 0.81-0.92) . However, a common cut-off point could not be determined . There was weak but significant correlation between DIAGNOdent readings and all three classes of lesion depth . Level of infection showed very weak correlation with the DIAGNOdent readings . It was concluded that DIAGNOdent is more reliable in detecting dentinal caries if a proper cut-off point is used than in indicating actual lesion depth . Readings from the different instruments were not directly comparable, however, owing to the lack of a common cut-off point.

J Vet Med Educ, 2004 Fall, 31(3), 216 - 21
Future directions in the European union for veterinary education as related to food-producing animals, with special reference to Greece; Kyriakis SC et al.; During the past 50 years, procedures for raising food-producing animals have changed . Intensification of food production was necessary to keep prices low and to fulfill market demands for the continuously increasing worldwide population . Intensification of farming procedures produced many new problems, some of which had a considerable impact on public opinion about how animals are raised and how food of animal origin is produced and preserved . "Man made diseases" of animals such as bovine spongiform encephalopathy (BSE); contamination of foods with dioxins either through contamination of animal feeds or from the environment; and increased microbial resistance to drugs used for treatment, for prophylaxis of animals from infectious agents, and for growth promotion are some well-known hazards of intensified farming . Veterinarians working on food-producing animals are faced with an increased demand for foods of high quality and safety in developed countries, and higher quantities in the rest of the world . These qualitative and quantitative changes indicate that they must adjust to these new conditions . They will be most successful if their education is adjusted to meet the challenges that the public has created for them through new concepts of the production of food of animal origin . One such concept is the production of foods under fully certified procedures from the farm to the consumer's table . Food safety measures protecting public health will better be achieved if the education of the future veterinarian includes the principles of Hazard Analysis Critical Control Points (HACCP) starting at farm level . This article provides some market-driven ideas in this direction for European Union (EU) countries, including Greece.

Panminerva Med, 2004 Sep, 46(3), 165 - 9
Is hepcidin the bridge linking Helicobacter pylori and anemia of chronic infection? A research proposal; Pellicano R et al.; Since the last decade, several studies have reported on the link between chronic Helicobacter pylori (H . pylori) or Helicobacter species (H . species) infection and a variety of extragastric manifestations, comprising iron-deficiency anemia . A crucial question concerns which possible pathogenic mechanism of H . pylori infection may be involved in chronic anemia . Recent findings support the hypothesis that in subjects with H . pylori-positive gastritis, concomitant changes in intragastric pH and ascorbic acid are present that might play a role in impairing alimentary iron absorption with consequent sideropenic anemia . It has also been speculated that H . pylori infected antrum could act as a sequestering focus for iron . The bacterium enhances gastric lactoferrin, which captures iron from transferrin . The iron thus bound to lactoferrin is in turn picked up by the bacterium, by means of its outer membrane receptors, for its own growth . These models, however, are not able to answer why iron-deficiency anemia does not develop in all infected subjects . Recently, a new anti-microbial liver-made peptide, namely hepcidin, has been characterised . The link between hepcidin induction, inflammation and anemia both in humans and in animal models supports its key role as mediator of anemia of inflammation . In the present paper, we highlight the data available on the association between H . pylori and iron-deficiency anemia and, we propose to evaluate a possible mechanism involving hepcidin in a bridging role linking the infection to the anemia.

Bioinformatics . 2004 Oct 27; {Epub ahead of print}
LabArray: real-time imaging and analytical tool for microarrays; Kok Ng JK et al.; SUMMARY: Microarrays have been used to perform high-throughput genetic analyses such as single-nucleotide polymorphisms detection and microbial genome analysis . Some of these analyses require real-time monitoring of the hybridisation signals with respect to a varying experimental condition, such as temperature . However, current microarray imaging and analysis packages typically do not possess such real-time capabilities . Therefore, microarray image analyses are often time-consuming and labour-intensive . LabArray was developed to expedite such processes by enabling real-time monitoring of microarray signals . AVAILABILITY: LabArray is available at SUPPLEMENTARY INFORMATION: Screenshots and instructions for use are available at the above website.

J Food Prot, 2004 Oct, 67(10), 2230 - 3
Effect of commonly used enhancement solutions on the viability of Toxoplasma gondii tissue cysts in pork loin; Hill DE et al.; Retail meat cuts of pork are frequently enhanced with salt solutions to improve flavor and texture and to extend shelf life through reductions in microbial contamination . A study of the effect of commonly used meat enhancement solutions on the viability of Toxoplasma gondii tissue cysts was performed using tissues from experimentally infected mice and pigs . Brains of T . gondii-infected mice were injected to 110% of the original weight of the brain with solutions containing sodium chloride (1 and 2%), sodium diacetate (0.1 and 0.2%), sodium tripolyphosphate (0.25 and 0.5%), potassium lactate (1.4 and 1.96%), or sodium lactate (1.4, 1.5, and 2.0%) alone or in combination and stored at 4 degrees C for 7 days before feeding to T . gondii-seronegative cats . Loins were collected from pigs experimentally infected with T . gondii and injected as above and stored for 7, 28, or 45 days at 4 degrees C before feeding to T . gondii-seronegative cats . Cat feces were examined for 14 days to assess oocyst shedding . The present study demonstrated that injection of mouse brains or pork loins with solutions containing 2% sodium chloride or > or =1.4% potassium or sodium lactate, alone or in combination with other components, prevented transmission of T . gondii to cats.

J Chromatogr A, 2004 Oct 1, 1050(2), 201 - 9
Detection and identification of phenazone-type drugs and their microbial metabolites in ground and drinking water applying solid-phase extraction and gas chromatography with mass spectrometric detection; Zuhlke S et al.; A new analytical method applying in situ derivatization was developed to enable the extraction of polar drug metabolites from water samples by solid-phase extraction (SPE) . An additional derivatization by silylation was used to enhance the sensitivity of analyte detection by gas chromatography-mass spectrometry (GC-MS) . Thus, the two metabolites 1,5-di-methyl-1,2-dehydro-3-pyrazolone (DP) and 4-(2-methylethyl)-1,5-dimethyl-1,2-dehydro-3-pyrazolone (PDP), postulated for the degradation of phenazone and propyphenazone, were identified and detected up to the microg/L level in raw and drinking water samples from public water supply.

Mol Nutr Food Res, 2004 Nov, 48(6), 434 - 40
Evaluation of the potential allergenicity of the enzyme microbial transglutaminase using the 2001 FAO/WHO Decision Tree; Pedersen MH et al.; All novel proteins must be assessed for their potential allergenicity before they are introduced into the food market . One method to achieve this is the 2001 FAO/WHO Decision Tree recommended for evaluation of proteins from genetically modified organisms (GMOs) . It was the aim of this study to investigate the allergenicity of microbial transglutaminase (m-TG) from Streptoverticillium mobaraense . Amino acid sequence similarity to known allergens, pepsin resistance, and detection of protein binding to specific serum immunoglobulin E (IgE) (RAST) have been evaluated as recommended by the decision tree . Allergenicity in the source material was thought unlikely, since no IgE-mediated allergy to any bacteria has been reported . m-TG is fully degraded after 5 min of pepsin treatment . A database search showed that the enzyme has no homology with known allergens, down to a match of six contiguous amino acids, which meets the requirements of the decision tree . However, there is a match at the five contiguous amino acid level to the major codfish allergen Gad c1 . The potential cross reactivity between m-TG and Gad c1 was investigated in RAST using sera from 25 documented cod-allergic patients and an extract of raw codfish . No binding between patient IgE and m-TG was observed . It can be concluded that no safety concerns with regard to the allergenic potential of m-TG were identified.

Mol Nutr Food Res, 2004 Nov, 48(6), 413 - 23
Allergy assessment of foods or ingredients derived from biotechnology, gene-modified organisms, or novel foods; Poulsen LK; The introduction of novel proteins into foods carries a risk of eliciting allergic reactions in individuals sensitive to the introduced protein and a risk of sensitizing susceptible individuals . No single predictive test exists to perform a hazard assessment in relation to allergenic properties of newly expressed proteins in gene-modified organisms (GMOs) . Instead, performance of a weighted risk analysis based on the decision tree approach has been suggested . The individual steps of this analysis comprise sequence homology to known allergens, specific or targeted serum screens for immunoglobulin E (IgE) cross-reactions to known allergens, digestability studies of the proteins in simulated gastric and/or intestinal fluids, and animal studies . These steps are discussed and five examples of risk evaluation of GMOs or novel foods are presented . These include ice-structuring protein derived from fish, microbial transglutaminase, GMO-soybeans, amylase and the Nangai nut.

What Is Bioassay?, What Is Yeast?, What Is Cell Biology?, What Is Anthrax?, What Is Bioengineering?, o, Microorganisms, e, Microorganism, r, Microbiology, c, Bacteriology, n, Bacterium, a, Antibiotics, n, Cell suspensions, e, Prokaryotes, o, Escherichia coli, i, Bacteriophages, e, Yeasts, i, Gram positive, o, Streptococcal, n, Pseudomonas, n, Biodegradation, i, Staphylococcus aureus, i, Staphylococcus aureus, c, Schizosaccharomyces, n, Streptomycin, s, Pseudomonas aeruginosa, n, Corynebacter, s, Enterobacters, n, Bacteriophage, e, Bacillus, c, Escherichia coli, s, Yeasts

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005