J Clin Microbiol, 2002 Dec, 40(12), 4757 - 9 Evaluation of a plastic nonvented aerobic blood culture bottle for use with the BacT/ALERT microbial detection system; Snyder JW et al.; The current BacT/ALERT SA (BTA SA) aerobic blood culture bottle is made from glass, does not require venting, and contains a liquid emulsion sensor (LES) . Its performance has been shown to be equivalent to that of the vented standard aerobic culture bottle . A further-improved version of the BTA SA bottle, designated the BacT/ALERT plastic SA (BTA PSA) culture bottle, is made from clear plastic to prevent breakage, does not require venting, and contains a modified LES (LES 2) to reduce the possibility of false positives . The BTA PSA provides a practical alternative to the current glass version of this bottle . The plastic bottle is also comparable to the current glass bottle in transparency and growth performance and additionally minimizes the exposure to infectious agents due to glass bottle breakage. J Anim Physiol Anim Nutr (Berl), 2002 Oct, 86(9-10), 347 - 52 Determination of phytase activity in cereal grains by direct incubation; Zimmermann B et al.; In contrast to supplemented microbial phytases, considerably lower cereal phytase activities were found after application of an enzyme solution extracted in citrate/NaOH buffer (pH 5.5) for 1 h as compared with the direct incubation of the plant material . Differences between both methods obtained were 70% for wheat and spelt and 50% for barley and rye . The determination of phytase activity of a wheat sample by direct incubation was affected by the sample size and amount of substrate (Na-phytate) added . Optimal conditions are a maximal level of phytase activity of < or = 0.05 U incubated for 60 min in a buffered solution (citrate/NaOH, pH 5.5) with 10 ml of 32.6 mmol/l Na-phytate at 37 composite function C . Possible reasons for the differences between both methods and for the factors affecting phytase activity by direct incubation are discussed. J Agric Food Chem, 2002 Dec 4, 50(25), 7412 - 9 Identification of the epsilon-(gamma-glutamyl)lysine cross-linking sites in alpha-lactalbumin polymerized by mammalian and microbial transglutaminases; Lee DS et al.; To investigate the site specificity of two transglutaminases (TGases), that is, the enzymes from guinea pig liver (GTGase) and Streptoverticillium (MTGase), the acyl acceptor and donor sites in alpha-lactalbumin were determined . Alpha-lactalbumin was cross-linked in the presence of dithiothreitol by GTGase and MTGase for 15 and 30 min, respectively . Cross-linked alpha-lactalbumins by GTGase and MTGase were digested with lysylendopeptidase followed by the separation of the resulting peptides using reverse-phase HPLC . By the sequence analysis of the peptide fragments containing two N termini, which indicates the presence of cross-linked peptide, the lysine residues targeted by TGases were identified as follows: for GTGase, Lys16, Lys93, and Lys122; for MTGase, Lys5 . These peptide fragments were further digested by V8 protease . Separation and sequence analyses of the resultant peptides were performed to identify glutamine residue involved in cross-linking . It was found that Gln54 was cross-linked to lysine residues by GTGase and MTGase in common . It is suggested that the difference in the numbers of lysine residues targeted by GTGase and MTGase may be responsible for the difference in the polymerization process of alpha-lactalbumin between GTGase- and MTGase-catalyzed systems. Am J Clin Nutr, 2002 Dec, 76(6), 1317 - 25 Contribution of intestinal microbial lysine to lysine homeostasis is reduced in minipigs fed a wheat gluten-based diet; Backes G et al.; BACKGROUND: We previously reported microbial lysine contribution to plasma lysine homeostasis in humans with an adequate lysine intake . OBJECTIVE: We sought to explore whether the low lysine intake from a wheat gluten-based diet is balanced by enhanced microbial lysine contribution in a pig model . DESIGN: Twenty miniature pigs (minipigs) fitted with ileo-ileal cannulas were fed 2 wheat gluten-based diets . One diet provided 2.7 g lysine/kg diet (WG diet) and one diet was supplemented with crystalline lysine to provide 6.6 g lysine/kg diet (WG+Lys diet) . Both diets were fed for 10 or 100 d (n = 5 per group): 10WG+Lys, 10WG, 100WG+Lys, and 100WG diets . Ileal microbial lysine, which we considered to be the precursor pool for absorption, was labeled by oral administration of (15)NH(4)Cl for the final 10 d . On days 10 and 100, a 10-h fast-fed tracer protocol with {1-(13)C}lysine was performed . RESULTS: Lysine rates of appearance decreased by 25% with the WG diet in the fed state but increased by 50% with the WG+Lys diet in the fasted state (P < 0.05) . Daily gross microbial lysine contribution was lower (P < 0.05) with the WG diet (205.3 micro mol . kg(-) (1) . d(-)(1)) than with the WG+Lys diet (370.7 micro mol . kg(-) (1) . d(-)(1)), irrespective of the adaptation period and was similar to the ileal lysine loss with the WG diet . In the WG groups, incorporation of microbial lysine increased in the duodenum and liver (P < 0.05) but not in whole-body and muscle proteins . CONCLUSION: Minipigs fed the WG diet did not adapt by showing an enhanced absorption of microbial lysine to the extrasplanchnic tissues, presumably because microbial lysine continues to be used for splanchnic protein synthesis. Astrobiology, 2002 Spring, 2(1), 83 - 92 Hydrogen in rocks: an energy source for deep microbial communities; Freund F et al.; To survive in deep subsurface environments, lithotrophic microbial communities require a sustainable energy source such as hydrogen . Though H2 can be produced when water reacts with fresh mineral surfaces and oxidizes ferrous iron, this reaction is unreliable since it depends upon the exposure of fresh rock surfaces via the episodic opening of cracks and fissures . A more reliable and potentially more voluminous H2 source exists in nominally anhydrous minerals of igneous and metamorphic rocks . Our experimental results indicate that H2 molecules can be derived from small amounts of H2O dissolved in minerals in the form of hydroxyl, OH- or O3Si-OH, whenever such minerals crystallized in an H2O-laden environment . Two types of experiments were conducted . Single crystal fracture experiments indicated that hydroxyl pairs undergo an in situ redox conversion to H2 molecules plus peroxy links, O3Si/OO\SiO3 . While the peroxy links become part of the mineral structure, the H2 molecules diffused out of the freshly fractured mineral surfaces . If such a mechanism occurred in natural settings, the entire rock column would become a volume source of H2 . Crushing experiments to facilitate the outdiffusion of H2 were conducted with common crustal igneous rocks such as granite, andesite, and labradorite . At least 70 nmol of H2/g diffused out of coarsely crushed andesite, equivalent at standard pressure and temperature to 5,000 cm3 of H2/m3 of rock . In the water-saturated, biologically relevant upper portion of the rock column, the diffusion of H2 out of the minerals will be buffered by H2 saturation of the intergranular water film. Astrobiology, 2001 Spring, 1(1), 111 - 23 Importance of a martian hematite site for astrobiology; Allen CC et al.; Defining locations where conditions may have been favorable for life is a key objective for the exploration of Mars . Of prime importance are sites where conditions may have been favorable for the preservation of evidence of prebiotic or biotic processes . Areas displaying significant concentrations of the mineral hematite (alpha-Fe2O3), recently identified by thermal emission spectrometry, may have significance in the search for evidence of extraterrestrial life . Since iron oxides can form as aqueous mineral precipitates, the potential exists to preserve microscopic evidence of life in iron oxide-depositing ecosystems . Terrestrial hematite deposits proposed as possible analogs for hematite deposits on Mars include massive (banded) iron formations, iron oxide hydrothermal deposits, iron-rich laterites and ferricrete soils, and rock varnish . We report the potential for long-term preservation of microfossils by iron oxide mineralization in specimens of the approximately 2,100-Ma banded iron deposit of the Gunflint Formation, Canada . Scanning and analytical electron microscopy reveals micrometer-scale rods, spheres, and filaments consisting predominantly of iron and oxygen with minor carbon . We interpret these objects as microbial cells permineralized by an iron oxide, presumably hematite . The confirmation of ancient martian microbial life in hematite deposits will require the return of samples to terrestrial laboratories . A hematite-rich deposit composed of aqueous iron oxide precipitates may thus prove to be a prime site for future sample return. Astrobiology, 2001 Spring, 1(1), 57 - 70 Microbial influences on local carbon isotopic ratios and their preservation in carbonate; Sumner DY; Analysis of the carbon isotopic composition of carbonates is a valuable tool for studying microbial processes and looking for evidence of life . Microbial fixation of CO2 and conversion of organic carbon to CO2 can produce measurable delta 13C shifts in a microbial mat environment . Diffusion modeling demonstrates that substantial isotopic shifts can develop within the mat and in the diffusion boundary layer in the fluid when CO2 fixation is rapid and prolonged for several hours . Carbonates that precipitate during rapid CO2 fixation can preserve these microbially produced isotopic shifts . However, continued precipitation during intervals when respiration dominates or after the cessation of active microbial growth commonly dilutes autotrophic isotopic signatures . Thus, preserved isotopic signatures rarely reflect the magnitude of isotopic shifts within the mat . Interpretation of observed isotopic shifts in microbial mat carbonate depends on fully characterizing ambient delta 13C and eliminating other origins for isotopic shifts . The carbon isotopic composition of reservoirs can vary substantially, both on Earth and on other planets . Characterizing the reservoir composition and any changes through time is critical to evaluating microbially induced shifts . In addition, careful evaluation of non-microbial causes for shifts in isotopic composition is essential for a reliable interpretation . Complicating processes include recrystallization, calcite precipitation over extended periods of time, variable precipitation rates and water chemistry, and mixing of carbonates having different isotopic signatures. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 599 - 606 Exploitation of genetically modified inoculants for industrial ecology applications; Morrissey JP et al.; The major growth seen in the biotechnology industry in recent decades has largely been driven by the exploitation of genetic engineering techniques . The initial benefits have been predominantly in the biomedical area, with products such as vaccines and hormones that have received broad public approval . In the environmental biotechnology and industrial ecology sectors, biotechnology has the potential to make significant advances through the use of genetically modified (GM) microbial inoculants that can reduce agri-chemical usage or remediate polluted environments . Although many GM inoculants have been developed and tested under laboratory conditions, commercial exploitation has lagged behind . Here, we review scientific and regulatory requirements that must be satisfied as part of that exploitation process . Particular attention is paid to new European Union (EU) regulations (Directives) that govern the testing and release of genetically modified organisms and microbial plant protection inoculants in the EU . With regard to the release of GM inoculants, the impact of the inoculant and the fate of modified genes are important concerns . Long term monitoring of release sites is necessary to address these issues . Data are reported from the monitoring of a site 6 years after release of GM Sinorhizobium meliloti strains . It was found that despite the absence of a host plant, the GM strains persisted in the soil for at least 6 years . Horizontal transfer and microevolution of a GM plasmid between S . meliloti strains was also observed . These data illustrate the importance of assessing the long-term persistence of GM inoculants. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 575 - 85 Comparative ecology of H2 cycling in sedimentary and phototrophic ecosystems; Hoehler TM et al.; The simple biochemistry of H2 is critical to a large number of microbial processes, affecting the interaction of organisms with each other and with the environment . The sensitivity of each of these processes to H2 can be described collectively, through the quantitative language of thermodynamics . A necessary prerequisite is to understand the factors that, in turn, control H2 partial pressures . These factors are assessed for two distinctly different ecosystems . In anoxic sediments from Cape Lookout Bight (North Carolina, USA), H2 partial pressures are strictly maintained at low, steady-state levels by H2-consuming organisms, in a fashion that can be quantitatively predicted by simple thermodynamic calculations . In phototrophic microbial mats from Baja California (Mexico), H2 partial pressures are controlled by the activity of light-sensitive H2-producing organisms, and consequently fluctuate over orders of magnitude on a daily basis . The differences in H2 cycling can subsequently impact any of the H2-sensitive microbial processes in these systems . In one example, methanogenesis in Cape Lookout Bight sediments is completely suppressed through the efficient consumption of H2 by sulfate-reducing bacteria; in contrast, elevated levels of H2 prevail in the producer-controlled phototrophic system, and methanogenesis occurs readily in the presence of 40 mM sulfate. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 487 - 507 Characterizing man-made and natural modifications of microbial diversity and activity in coastal ecosystems; Paerl HW et al.; The impacts of growing coastal pollution and habitat alteration accompanying human encroachment are of great concern at the microbial level, where much of the ocean's primary production and biogeochemical cycling takes place . Coastal ecosystems are also under the influence of natural perturbations such as major storwns and flooding . Distinguishing the impacts of natural and human stressors is essential for understanding environmentally-induced change in microbial diversity and function . The objective of this paper is to discuss the applications and merits of recently developed molecular, ecophysiological and analytical indicators and their utility in examining anthropogenic and climatic impacts on the structure and function of coastal microbial communities . The nitrogen-limited Neuse River Estuary and Pamlico Sound, North Carolina are used as examples of ecosystems experiencing both anthropogenic (i.e., accelerating eutrophication) and climatic stress (increasing frequencies of tropical storms and hurricanes) . Additional examples are derived from a coastal monitoring site (LEO) on the Atlantic coast of New Jersey and Galveston Bay, on the Gulf of Mexico . In order to assess structure, function, and trophic state of these and other coastal ecosystems, molecular (DNA and RNA-based) characterizations of the microbial taxa involved in carbon, nitrogen and other nutrient transformations can be combined with diagnostic pigment-based indicators of primary producer groups . Application of these methods can reveal process-level microbial community responses to environmental variability over a range of scales . Experimental approaches combined with strategic monitoring utilizing these methods will facilitate: (a) understanding organismal and community responses to environmental change, and (b) synthesizing these responses in the context of ecosystem models that integrate physical, chemical and biotic variability with environmental controls. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 309 - 18 Bacteria and protozoa as integral components of the forest ecosystem--their role in creating a naturally varied soil fertility; Clarholm M; The paper explores interactions between the two first organism groups to appear on earth, the bacteria and protozoa, and their interplay with the rest of the ecosystem focusing upon northern boreal forests . The microbial loop is suggested as a mechanism for local inputs of new N to the ecosystem . The possibility to couple short-term microbial processes with their long-term effects,--as registered in plants, soil and the atmosphere, via the abiotic variables--is explored . The latter are investigated in relation to the environments they create for the micro-organisms, and how this results in varying soil fertility . A chain of events is presented that relate high Ca concentration in the mineral soil and high water availability to increased nitrogen availability for plants via the micro-organisms . An example is given of the influence of these parameters directly upon protozoa along an extreme fertility gradient, and also indirect evidence from a Finnish field study of 30 sites with four fertility levels . Finally, there is a discussion about ways to convert knowledge gained in detailed studies of microbial interactions into forms useful when evaluating the present status of and effects of ameliorative management on ecosystems strongly affected by humans. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 181 - 7 The distribution and activity of sulphate reducing bacteria in estuarine and coastal marine sediments; Purdy KJ et al.; Sulphate-reducing bacteria (SRB) play a vital role both the carbon and sulphur cycles and thus are extremely important components of the global microbial community . However, it is clear that the ecology, the distribution and activity of different SRB groups is poorly understood . Probing of rRNA suggests that different sediments have distinctly different patterns of SRB with complex factors controlling the activity of these organisms . The linking of community structure and function using sediment slurry microcosms suggests that certain groups of SRB, e.g., Desulfobacter and Desulfobulbus, can be linked to the use of specific substrates in situ . However, it is still unclear what environmental substrates are utilised by the majority of known SRBs . The work to date has greatly enhanced our understanding of the ecology of these organisms and is beginning to suggest patterns in their distribution and activity that may be relevant to understanding microbial ecology in general. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 155 - 64 Experimental social evolution with Myxococcus xanthus; Velicer GJ et al.; Genetically-based social behaviors are subject to evolutionary change in response to natural selection . Numerous microbial systems provide not only the opportunity to understand the genetic mechanisms underlying specific social interactions, but also to observe evolutionary changes in sociality over short time periods . Here we summarize experiments in which behaviors of the social bacterium Myxococcus xanthus changed extensively during evolutionary adaptation to two relatively asocial laboratory environments . M . xanthus moves cooperatively, exhibits cooperative multicellular development upon starvation and also appears to prey cooperatively on other bacteria . Replicate populations of M . xanthus were evolved in both structured (agar plate) and unstructured (liquid) environments that contained abundant resources . The importance of social cooperation for evolutionary fitness in these habitats was limited by the absence of positive selection for starvation-induced spore production or predatory efficiency . Evolved populations showed major losses in all measured categories of social proficiency- motility, predation, fruiting ability, and sporulation . Moreover, several evolved genotypes were observed to exploit the social behavior of their ancestral parent when mixed together during the developmental process . These experiments that resulted in both socially defective and socially exploitative genotypes demonstrate the power of laboratory selection experiments for studying social evolution at the microbial level . Results from additional selection experiments that place positive selection pressure on social phenotypes can be integrated with direct study of natural populations to increase our understanding of principles that underlie the evolution of microbial social behavior. Water Res, 2002 Nov, 36(19), 4916 - 26 Role of iron in controlling speciation and mobilization of arsenic in subsurface environment; Bose P et al.; Widespread arsenic contamination of groundwater has been reported of late in Bangladesh and West Bengal state of India . On the basis of arsenic geochemistry, three probable mechanisms have been cited for arsenic mobility in aquifers of West Bengal and Bangladesh . First, mobilization of arsenic due to the oxidation of arsenic-bearing pyrite minerals . Second, dissolution of arsenic-contaminated iron oxy-hydroxides (FeOOH) due to onset of reducing conditions in the subsurface . Third, due to the release of arsenic sorbed to aquifer minerals by competitive exchange with phosphate ions, that migrates into aquifers due to application of fertilizer to surface soil . Based on the review of field data from the affected region, it appears that the second mechanism described above is the most probable . Two reduction processes associated with this mechanism were investigated, viz., reduction of iron oxy-hydroxide to iron (II), which results in the mobilization of arsenic, and reduction of arsenic (V) to arsenic (III), which may enhance mobility of arsenic under certain conditions . These reactions, in the opinion of some researchers, are possible in subsurface environments mainly through microbial intervention . However, through the data presented in this paper, it has been demonstrated that above red-ox reactions involving iron and arsenic are also possible through predominantly abiotic pathways . While these results do not necessarily imply that abiotic red-ox processes are dominant in all subsurface environments containing iron and arsenic, it is entirely possible that abiotic interactions as described here may be responsible for a substantial amount of transformations involving iron and arsenic in anoxic subsurface environments. Water Res, 2002 Nov, 36(19), 4884 - 92 Effect of alum treatment on the trihalomethane formation and bacterial regrowth potential of natural and synthetic waters; Page DW et al.; Waters from five reservoirs and "synthetic waters", prepared using terrestrially derived dissolved organic matter (DOM) extracted from vegetation and reservoir catchment soils, were studied for their treatability with alum using a jar test procedure . DOM in drinking water is a precursor for the formation of trihalomethanes (THM) following chlorine disinfection and can also be a substrate for microbial growth in the drinking water distribution system . The trihalomethane formation potential (THMFP) represents an upper concentration limit on THMs formed by chlorination, while bacterial regrowth potential (BRP) is an indicator of the bioavailability of DOM . BRP and THMFP were measured before and after alum treatment and the results were related to the source of the DOM . It was found that freshly derived terrestrial DOM in synthetic water resulted in higher THMFP and BRP than DOM in reservoir waters . For the samples investigated, conventional alum treatment did not always reduce the THM precursor levels formed in laboratory tests below the NH&MRC (1996) guideline level of 250 microg/L nor produce microbially stable waters. Water Res, 2002 Nov, 36(19), 4785 - 94 Atrazine mineralization potential in two wetlands; Anderson KL et al.; The fate of atrazine in agricultural soils has been studied extensively but attenuation in wetland systems has received relatively little attention . The purpose of this study was to evaluate the mineralization of atrazine in two wetlands in central Ohio . One was a constructed wetland, which is fed by Olentangy River water from an agricultural catchment area . The other was a natural fen (Cedar Bog) in proximity to atrazine-treated cornfields . Atrazine mineralization potential was measured by 14CO2 evolution from {U-ring-14C}-atrazine in biometers . The constructed wetland showed 70-80% mineralization of atrazine within 1 month . Samples of wetland water that were pre-concentrated 200-fold by centrifugation also mineralized 60-80% of the added atrazine . A high extent of atrazine mineralization (75-81% mineralized) was also associated with concentrated water samples from the Olentangy River that were collected upstream and downstream of the wetland . The highest levels of mineralization were localized to the top 5 cm zone of the wetland sediment, and the activity close to the outflow at the Olentangy wetland was approximately equal to that near the inflow . PCR amplification of DNA extracted from the wetland sediment samples showed no positive signals for the atzA gene (atrazine chlorohydrolase), while Southern blots of the amplified DNA showed positive bands in five of the six Olentangy wetland sediment samples . Amplification with the trzD (cyanuric acid amidohydrolase) primers showed a positive PCR signal for all Olentangy wetland sediment samples . There was little mineralization of atrazine in any of the Cedar Bog samples . DNA extracted from Cedar Bog samples did not yield PCR products, and the corresponding Southern hybridization signals were absent . The data show that sediment microbial communities in the Olentangy wetland mineralize atrazine . The level of activity may be related to the seasonality of atrazine runoff entering the wetland . Comparable activity was not observed in the Cedar Bog, perhaps because it does not directly receive agricultural runoff . Qualitatively, the detection of the genes was associated with measurable mineralization activity which was consistent with the differences between the two study sites. Water Sci Technol, 2002, 46(9), 109 - 15 Comparative study on microbial removal in immersed membrane filtration (IMF) with and without powdered activated carbon (PAC); Ujang Z et al.; This paper describes an investigation on the effect of microbial removal using IMF for high quality drinking water production . The comparison of IMF and IMF-PAC configuration was carried out in the study to highlight the importance of PAC in the system . The specific objective of this study was to study the effect of PAC adsorption in the IMF-PAC system particularly in removing microbial substances from contaminated raw water . A bench scale IMF-PAC configuration using a flat sheet microfiltration membrane was set up for experimental purposes . Experimentally, the result has shown high removal of microbial substances with the IMF-PAC system compared to IMF . The result of E . coli removal achieved was below the detectable level due to the microbial size, which is bigger than membrane pore size . The addition of PAC has shown a direct effect on total microbial removal . The adsorption of microbial onto PAC surfaces reduced the amount of smaller microbial present in permeate samples . As a conclusion, the configuration of IMF is a promising separation process in removing microbial substances, especially when the system is combined with PAC. Microb Ecol, 2003 Jan, 45(1), 11 - 9 Epub 2002 Nov 27. Effect of inorganic nutrients on relative contributions of fungi and bacteria to carbon flow from submerged decomposing leaf litter; Gulis V et al.; The relative contributions of fungi and bacteria to carbon flow from submerged decaying plant litter at different levels of inorganic nutrients (N and P) were studied . We estimated leaf mass loss, fungal and bacterial biomass and production, and microbial respiration and constructed partial carbon budgets for red maple leaf disks precolonized in a stream and then incubated in laboratory microcosms at two levels of nutrients . Patterns of carbon flow for leaf disks colonized with the full microbial assemblage were compared with those colonized by bacteria but in which fungi were greatly reduced by placing leaf disks in colonization chambers sealed with membrane filters to exclude aquatic hyphomycete conidia but not bacterial cells . On leaves colonized by the full microbial assemblage, elevated nutrient concentrations stimulated fungi and bacteria to a similar degree . Peak fungal and bacterial biomass increased by factors of 3.9 and 4.0; cumulative production was 3.9 and 5.1 times higher in the high nutrient in comparison with the low nutrient treatment, respectively . Fungi dominated the total microbial biomass (98.4 to 99.8%) and cumulative production (97.3 and 96.5%), and the fungal yield coefficient exceeded that of bacteria by a factor of 36 and 27 in low- and high-nutrient treatments, respectively . Consequently, the dominant role of fungi in leaf decomposition did not change as a result of nutrient manipulation . Carbon budgets indicated that 8% of leaf carbon loss in the low-nutrient treatment and 17% in the high-nutrient treatment were channeled to microbial (essentially fungal) production . Nutrient enrichment had a positive effect on rate of leaf decomposition only in microcosms with full microbial assemblages . In treatments where fungal colonization was reduced, cumulative bacterial production did not change significantly at either nutrient level and leaf decomposition rate was negatively affected (high nutrients), suggesting that bacterial participation in carbon flow from decaying leaf litter is low regardless of the presence of fungi and nutrient availability . Moreover, 1.5 and 2.3 times higher yield coefficients of bacteria in the reduced fungal treatments at low and high nutrients, respectively (percentage of leaf carbon loss channeled to bacterial production), suggest that bacteria are subjected to strong competition with fungi for resources available in leaf litter. Microb Ecol, 2003 Jan, 45(1), 29 - 38 Epub 2002 Nov 27. gfp-Tagged cells as a useful tool to study the survival of Escherichia coli in the presence of the river microbial community; Arana I et al.; We have used an Escherichia coli strain DH5a containing pGreenTIR to study the survival of this bacterium in river water . As green fluorescence was maintained throughout survival both in dark and illuminated conditions, gfp-tagged E . coli cells were clearly distinguished from the microbial community of the river Butron . gfp-tagged E . coli cells were monitored to estimate total density as well as the density of the culturable and viable (active electron transport system, CTC+) cells . Our results indicate that autochthonous bacteria and introduced E . coli are predated by flagellates . The autochthonous bacterial community behaves as predation-escaping prey, showing a tendency to cellular miniaturization and so maintaining the density of the population . In contrast, introduced E . coli behaves as predation-non-escaping prey, so E . coli was eliminated from the system . When comparing the elimination by predation of heat-treated and non-heated gfp-tagged E . coli cells we deduce that the flagellates do not discriminate between live and heat-treated cells . Finally, in the presence of the river microbial community, the E . coli cells appeared to be ingested before cellular deterioration could occur . Thus predation reduces the quantitative importance of the viable but nonculturable (VBNC) population of E . coli in the aquatic systems. Proc Natl Acad Sci U S A, 2002 Dec 10, 99(25), 16144 - 9 Epub 2002 Nov 21. Characteristic genome rearrangements in experimental evolution of Saccharomyces cerevisiae; Dunham MJ et al.; Genome rearrangements, especially amplifications and deletions, have regularly been observed as responses to sustained application of the same strong selective pressure in microbial populations growing in continuous culture . We studied eight strains of budding yeast (Saccharomyces cerevisiae) isolated after 100-500 generations of growth in glucose-limited chemostats . Changes in DNA copy number were assessed at single-gene resolution by using DNA microarray-based comparative genomic hybridization . Six of these evolved strains were aneuploid as the result of gross chromosomal rearrangements . Most of the aneuploid regions were the result of translocations, including three instances of a shared breakpoint on chromosome 14 immediately adjacent to CIT1, which encodes the citrate synthase that performs a key regulated step in the tricarboxylic acid cycle . Three strains had amplifications in a region of chromosome 4 that includes the high-affinity hexose transporters; one of these also had the aforementioned chromosome 14 break . Three strains had extensive overlapping deletions of the right arm of chromosome 15 . Further analysis showed that each of these genome rearrangements was bounded by transposon-related sequences at the breakpoints . The observation of repeated, independent, but nevertheless very similar, chromosomal rearrangements in response to persistent selection of growing cells parallels the genome rearrangements that characteristically accompany tumor progression. J Invertebr Pathol, 2002 Oct, 81(2), 78 - 85 Sporulation of Metarhizium anisopliae and Beauveria bassiana on Coptotermes formosanus and in vitro; Sun J et al.; The sporulation of 22 total isolates of Metarhizium anisopliae and Beauveria bassiana was quantified on cadavers of the Formosan subterranean termite, Coptotermes formosanus . Conidial production increased significantly over 11 days post-death . Effects of isolates of M . anisopliae and B . bassiana on in vivo sporulation were significant . Although the overall effects of fungal species on in vivo sporulation were not significant, the interactions between fungal species and certain times post-death were significant, indicating different sporulation patterns between the two fungal species . B . bassiana isolates could be categorized into a group with high total sporulation (day 11) and low quick sporulation (on days 2 and 3), while M . anisopliae isolates fell into another group with high quick sporulation and low total sporulation . This could give M . anisopliae an advantage over B . bassiana in termite microbial control due to termite defensive social behaviors . Conidial production was significantly higher in vitro than in vivo . In vitro and in vivo sporulation differed by as much as 89x and 232x among the selected isolates of B . bassiana and M . anisopliae, respectively . Correlation between in vivo and in vitro conidial production was positive and significant . This may allow preliminary in vitro screening of a large number of isolates for high in vivo sporulation. Biosens Bioelectron, 2003 Jan, 18(1), 23 - 9 Development and characterization of a novel immobilized microbial membrane for rapid determination of biochemical oxygen demand load in industrial waste-waters; Rastogi S et al.; The rapid determination of waste-water quality of waste-water treatment plants in terms of pollutional strength, i.e . biochemical oxygen demand (BOD) is difficult or even impossible using the chemical determination method . The present study reports the determination of BOD within minutes using microbial BOD sensors, as compared to the 5-day determination using the conventional method . Multiple criteria establish the basis for the development of a BOD biosensor useful for rapid and reliable BOD estimation in industrial waste-waters . Of these, preparation of a suitable novel immobilized microbial membrane used in conjunction with an apt transducer is discussed . As a result, a microbial biosensor based on a formulated, synergistic, pre-tested microbial consortium has been developed for the measurement of BOD load of various industrial waste-waters . The sensor showed maximum response in terms of current difference, when a cell concentration of 2.25 x 10(10) CFU, harvested in their log phase of growth were utilized for microbial membrane construction . The sensor showed a stability of 180 days when the prepared membranes were stored at a temperature of 4 degrees C in 50 mM phosphate buffer of pH 6.8 . The reusability of the immobilized membranes was up to 200 cycles without appreciable loss of their response characteristics . A linear relationship between the current change and a glucose-glutamic acid (GAA) concentration up to 60 mg l(-1) was observed (r=0.999) . The lower detection limit was 1.0 mg l(-1) BOD . The sensor response was reproducible within +/-5% of the mean in a series of ten samples having 44 mg l(-1) BOD using standard a GGA solution . When used for the BOD estimation of industrial waste-waters, a relatively good agreement was found between the two methods, i.e . 5-day BOD and that measured by the developed microbial sensor. J Invest Dermatol, 2002 Nov, 119(5), 1096 - 102 Plasmacytoid dendritic cells: a new cutaneous dendritic cell subset with distinct role in inflammatory skin diseases; Wollenberg A et al.; Epidermal dendritic cells found in inflamed skin include Langerhans cells and the recently identified population of inflammatory dendritic epidermal cells . Another subset of dendritic cells in humans is the plasmacytoid dendritic cell in peripheral blood, which is characterized by the production of large amounts of type I interferon (interferon-alpha and interferon-beta) upon viral infection . We hypothesized that plasmacytoid dendritic cells might be involved in anti-viral defense mechanisms of the skin . Here we investigated plasmacytoid dendritic cells, inflammatory dendritic epidermal cells, and Langerhans cells in epidermal single cell suspensions of normal looking skin from healthy volunteers and of lesional skin from patients with different inflammatory skin diseases . Langerhans cells were found in normal and in inflamed skin samples . In normal skin, plasmacytoid dendritic cells and inflammatory dendritic epidermal cells were low or absent . Lesional skin samples from patients with psoriasis vulgaris and contact dermatitis contained relatively high numbers of both inflammatory dendritic epidermal cells and plasmacytoid dendritic cells . In contrast, many inflammatory dendritic epidermal cells but only very few plasmacytoid dendritic cells could be detected in atopic dermatitis lesions . Lupus erythematosus was characterized by high numbers of plasmacytoid dendritic cells but low numbers of inflammatory dendritic epidermal cells . These results demonstrate that in addition to resident Langerhans cells, plasmacytoid dendritic cells and inflammatory dendritic epidermal cells are selectively recruited to the skin lesions depending on the type of skin disease . The lack of plasmacytoid dendritic cells in atopic dermatitis may predispose atopic dermatitis patients to viral infections such as eczema herpeticum, a secondary infection of atopic dermatitis lesions with herpes simplex virus . The composition of dendritic cell subsets may help to clarify the etiology of inflammatory skin diseases and forms the basis for therapeutic intervention with selective microbial molecules such as immunostimulatory CpG oligonucleotides. Eur J Biochem, 2002 Dec, 269(23), 5963 - 71 Guanosine diphosphate-4-keto-6-deoxy-d-mannose reductase in the pathway for the synthesis of GDP-6-deoxy-d-talose in Actinobacillus actinomycetemcomitans; Suzuki N et al.; The serotype a-specific polysaccharide antigen of Actinobacillus actinomycetemcomitans is an unusual sugar, 6-deoxy-d-talose . Guanosine diphosphate (GDP)-6-deoxy-d-talose is the activated sugar nucleotide form of 6-deoxy-d-talose, which has been identified as a constituent of only a few microbial polysaccharides . In this paper, we identify two genes encoding GDP-6-deoxy-d-talose synthetic enzymes, GDP-alpha-d-mannose 4,6-dehydratase and GDP-4-keto-6-deoxy-d-mannose reductase, in the gene cluster required for the biosynthesis of serotype a-specific polysaccharide antigen from A . actinomycetemcomitans SUNYaB 75 . Both gene products were produced and purified from Escherichia coli transformed with plasmids containing these genes . Their enzymatic reactants were analysed by reversed-phase HPLC (RP-HPLC) . The sugar nucleotide produced from GDP-alpha-d-mannose by these enzymes was purified by RP-HPLC and identified by electrospray ionization-MS, 1H nuclear magnetic resonance, and GC/MS . The results indicated that GDP-6-deoxy-d-talose is produced from GDP-alpha-d-mannose . This paper is the first report on the GDP-6-deoxy-d-talose biosynthetic pathway and the role of GDP-4-keto-6-deoxy-d-mannose reductase in the synthesis of GDP-6-deoxy-d-talose. J Nat Prod, 2002 Nov, 65(11), 1693 - 5 Microbial metabolism of artemisinin by Mucor polymorphosporus and Aspergillus niger; Zhan JX et al.; Artemisinin (1) was transformed by Mucor polymorphosporus and Aspergillus niger . Five products were identified as 9beta-hydroxyartemisinin (2), 3beta-hydroxyartemisinin (3), deoxyartemisinin (4), 3beta-hydroxydeoxyartemisinin (5), and 1alpha-hydroxydeoxyartemisinin (6) . Products 2, 3, and 6 are new compounds. J Immunol, 2002 Dec 1, 169(11), 6210 - 7 Inhibition of human CD4(+)CD25(+high) regulatory T cell function; Baecher-Allan C et al.; CD4(+)CD25(+high) T cells are potent regulators of autoreactive T cells . However, it is unclear how regulatory CD4(+)CD25(+high) cells discriminate between desirable inflammatory immune responses to microbial Ags and potentially pathologic responses by autoreactive T cells . In this study, an in vitro model was created that allowed differential activation of regulatory CD4(+)CD25(+high) and responder CD4(+) T cells . If CD4(+)CD25(+high) regulatory cells were strongly activated, they maintained suppressive effector function for only 15 h, while stimulation with weaker TCR stimuli produced regulatory cells that were suppressive until 60 h after activation . In contrast, strongly activated CD4(+) responder T cells were resistant to regulation at all time points, while weakly stimulated CD4(+) cells were sensitive to suppression until 38 or 60 h after activation depending upon the strength of the stimulus . The extent of suppression mediated by CD4(+)CD25(+high) cells also depended on the strength of stimulation in an Ag-specific system . Thus, the stronger the TCR signal, the more rapidly and more completely the responder cells become refractory to suppression. Rapid Commun Mass Spectrom, 2002, 16(23), 2172 - 8 Use of labelled nitrogen to measure gross and net rates of mineralization and microbial activity in permanent pastures following fertilizer applications at different time intervals; Hatch DJ et al.; Measurements of some of the main internal N-cycling processes in soil were obtained by labelling the inorganic N pool with the stable isotope of nitrogen ((15)N) . The (15)N mean pool dilution technique, combined with other field measurements, enabled gross and net N-mineralization rates to be resolved in grassland soils, which had previously either received fertilizer N (F), or had remained unfertilized (U) for many years . The two soils were subdivided into plots that received N at different time intervals (over 3 weeks), prior to (15)N measurements being made . By this novel approach, possible 'priming' effects over time were investigated to try to overcome some of the temporal problems of isotopic labelling of soil N (native plus fertilizer) and to identify possible changes in a range of primary N-transformation processes . The results suggested that an overall stimulation of microbially mediated processes occurred with all N treatments, but there were inconsistencies associated with the release of N, both in the timing and the degree to which different processes responded to the application of fertilizer N . The rates of these processes were, however, within the range of previously reported data and the (15)N measurements were not adversely affected by the differences in N pools created by the treatments . Thus, the mean pool dilution technique was shown to be applicable to agricultural soils, under conditions relevant to grass swards receiving fertilizer . For example, between the U and F treatments, the size of inorganic N pools increased by five-fold and gross rates of mineralization reached 3.5 and 4.8 microg N g(-1) (dry soil) d(-1), respectively, but did not vary greatly with the timing of N applications . A correlation (r(2) = 0.57) was found between soil respiration (which is relatively simple to measure) and net mineralization (which is more time consuming), suggesting that the former might be used as an indicator of the latter . Although this relationship was stronger in previously unfertilized soils, the similarities found with fertilized soils suggest that this approach could be used to obtain information of wider agronomic value and would, therefore, warrant further work under a range of soil conditions . Rapid Commun Mass Spectrom, 2002, 16(23), 2157 - 64 Characterization of soil organic matter fractions from grassland and cultivated soils via C content and delta13C signature; Accoe F et al.; Variations in (13)C natural abundance and distribution of total C among five size and density fractions of soil organic matter, water soluble organic C (WSOC) and microbial biomass C (MBC) were investigated in the upper layer (0-20 cm) of a continuous grassland soil (CG, C(3) vegetation), a C(3)-humus soil converted to continuous maize cultivation (CM, C(4) vegetation) and a C(3)-humus soil converted to a rotation of maize cultivation and grassland (R) . The amounts of WSOC and MBC were both significantly larger in the CG than in the CM and the R . In the three soils, WSOC was depleted while MBC was enriched in (13)C as compared with whole soil C . The relative contributions to the total C content of C stored in the macro-organic matter and in the size fraction 50-150 microm decreased with decreasing total C contents in the order CG > R > CM, while the relative contribution of C associated with the clay- and silt-sized fraction <50 microm increased . This reflects a greater stability and physical protection against microbial degradation associated with soil disruption (tillage) of the clay- and silt-associated organic C, in relation to the organic C in larger size fractions . The size and density fractions from the CG soil showed significant differences in (13)C enrichment, indicating different degrees of microbial degradation and stability of soil organic C associated with physically different soil organic matter (SOM) fractions . Delta(13)C analysis of the size and density fractions from CM and R soils reflected a decreasing turnover rate of soil organic C with increasing density among the macro-organic matter fractions and with decreasing particle size . Glycoconj J, 2001 Oct, 18(10), 827 - 34 An alpha-L-fucosidase from Thermus sp . with unusually broad specificity; Eneyskaya EV et al.; An alpha-L-fucosidase (E.C . 3.2.1.51) exhibiting a wide aglycon specificity expressed in ability of cleaving alpha1 --> 6-, alpha1 -->3-, alpha1 --> 4-, and alpha1 --> 2-O-fucosyl bonds in fucosylated oligosaccharides, has been isolated from culture filtrate of Thermus sp . strain Y5 . The alpha-L-fucosidase hydrolyzes p-nitrophenyl alpha-L-fucopyranoside with V(max) of 12.0 +/- 0.1 microM/min/mg and K(m) = 0.20 +/- 0.05 mM and is able to cleave off about 90% of total L-fucose from pronase-treated fractions of fucosyl-containing glycoproteins and about 30% from the native glycoproteins . The purified enzyme is a tetramer with a molecular mass of 240 +/- 10 kDa consisting of four identical subunits with a molecular mass of 61.0 +/- 0.5 kDa . The N-terminal sequence showed homology to some alpha-L-fucosidases from microbial and plant sources . Hydrolysis of p-nitrophenyl alpha-L-fucopyranoside occurs with retention of the anomeric configuration . Transglycosylating activity of the alpha-L-fucosidase was demonstrated in reactions with such acceptors as alcohols, N-acetylglucosamine and N-acetylgalactosamine while no transglycosylation products were observed in the reaction with p-nitrophenyl alpha-L-fucopyranoside . The enzyme can be classified in glycosyl hydrolase family 29. Curr Gastroenterol Rep, 2002 Dec, 4(6), 481 - 9 The pathogenesis of inflammatory bowel disease: translational implications for clinicians; Abreu MT; Research in the pathogenesis of inflammatory bowel disease (IBD) has dramatically broadened our understanding of these complex disorders . These clinical manifestations result from a dysregulated immune response in the presence of luminal bacteria . Recent identification of mutations in the NOD2 gene, a protein involved in the sensing of bacteria, offers genetic support for the model of perturbed host-microbial interactions in Crohn's disease . Several immunologic pathways have been identified that play a role in maintaining gut immune homeostasis . Abnormal expression of proinflammatory, deleterious cytokines such as tumor necrosis factor-a and interferon-g results in direct and indirect tissue damage . The search for specific causative microbial agents in IBD continues to be intense . This paper describes the advances in our understanding of IBD pathogenesis, with an emphasis on how this information is translated into patient care . The next stage of research will take advantage of such molecular biologic techniques to identify new pathogenetic mechanisms and targets for therapy tailored to individual patients. Herz, 2002 Nov, 27(7), 669 - 76 Pathophysiology of cardiac inflammation: molecular mechanisms; Pankuweit S et al.; BACKGROUND: Inflammatory processes induced by rival infection are believed to be one of the major pathogenetic mechanisms in inflammatory dilated cardiomyopathy . Although the reason for progression to myocardial failure is not fully understood, postulated mechanisms include persistent viral infection alone or in combination with autoimmune processes . PATHOPHYSIOLOGY: Murine models of myocarditis have provided insight into the mechanisms by which autoimmune responses to cardiac antigens, probably in response to viral infection of the myocardium, arise and cause tissue pathology . Organ-specificity, cross-reactivity between microbial agents and cardiac tissue, and induction of tolerance to self-antigen are issues still at stake . In addition, cytokines mediate activation and effector phase of innate and specific immunity, which are both important in controlling viral infection . The innate immune response not only has an important protective function but also serves to initiate and regulate subsequent specific immune responses . In man, on the one hand specific T cells and antibodies against different cardiac tissue components have been demonstrated in myocardium and sera of patients with inflammatory cardiomyopathy, and on the other hand viral genome has been identified in endomyocardial biopsies due to the rapid development of new molecular biological techniques such as polymerase chain reaction (PCR), southern blot analysis and in-situ hybridization . But it is still a mater of debate whether virus infection itself, the ensuing immune response, or both, contribute to the deterioration of left ventricular function . CONCLUSION: Taking these mechanisms into account, screening for viral genome by PCR and detection of inflammatory infiltrates by immunohistochemistry are considered crucial for the establishment of a definite diagnosis thereby allowing for the initiation of specific therapeutic strategies. Res Microbiol, 2002 Oct, 153(8), 487 - 91 The subsurface origin of microbial life on the Earth; Trevors JT; Life on Earth can be divided into life on the surface made possible by photosynthesis and subsurface life with chemical energy as the driving force . An understanding of both environments is central to our understanding of the origin of life, the search for novel microbial species in the subsurface and for extraterrestrial life or life signatures . In this manuscript, the surface and subsurface worlds are examined with a focus on the origin or assembly of bacterial life. Immunobiology, 2002 Oct, 206(4), 392 - 407 Interaction of lipoteichoic acid and CpG-DNA during activation of innate immune cells; Dalpke AH et al.; The innate immune system recognizes pathogen-associated molecular patterns (PAMP) to cope with evolving infections . Toll-like receptors (TLRs) play a pivotal role in recognition of PAMPs . In the course of infection not a single but rather a full panel of different microbial components interacts with distinct TLRs simultaneously . Only limited information is available on effects of combinations of TLR agonists . Here, we have analyzed the effects of lipoteichoic acid (LTA), CpG-DNA and combinations thereof on innate immune cells in vitro . Although proinflammatory cytokines like TNF-alpha were induced by these agonists in quite similar amounts, CpG DNA was superior in its potency to induce IL-12p40 reflecting important differences in the biological valence of LTA and CpG-DNA . When given in combination, LTA and CpG-DNA were additive in induction of TNF-alpha, IL-6 and nitric oxide in RAW 264 macrophages, peritoneal macrophages and dendritic cells . Additive effects were also observed in regard to TNF-alpha mRNA . In contrast, LTA suppressed IL12p40 secretion induced by CpG-DNA in RAW cells and peritoneal macrophages but not in dendritic cells . Intracellular signal cascades (NFkappaB and p38 MAP kinase) showed additive effects after simultaneous triggering . mRNA expression ofTLRs showed only minor regulation after CpG or LTA application and thus does not account for the additive/suppressive effects observed . These results indicate that the consequences of interaction of innate immune cells with microbial pattern depend on the responding cell type and might be differential for certain effector mechanisms . Thus, the pathogen-characteristic panel of TLR ligands will induce pathogen-specific innate responses decisive for the inflammatory reactions. Microbiol Immunol, 2002, 46(9), 593 - 9 Effects of recombinant cholera toxin B subunit on IL-1beta production by macrophages in vitro; Maeyama J et al.; Recombinant cholera toxin B subunit (rCTB) is a safe and potent mucosal adjuvant . As a clue to the mechanism of the adjuvant effect of rCTB, the profile of cytokines secreted in vitro by the mouse peritoneal macrophage (Mphi) treated with rCTB was examined . IL-1beta secretion, intracellular production, and expression of its mRNA of LPS-stimulated Mphi was greatly enhanced by treatment with rCTB . IL-1beta production in response to other microbial stimulators, such as Pansorbin, Sansorbin, insoluble peptidoglycan, and Taxol, was also potentiated by rCTB . Mphi pretreated with rCTB before 24 hr could maintain the ability to produce a high level of IL-1beta, suggesting that this ability may be involved in the adjuvant activity of rCTB on Mphi stimulation . The possibility of close association between rCTB and signal transduction of a Toll-like receptor family in Mphi is discussed. Amino Acids, 2002, 23(4), 419 - 26 Effect of taurine chloramine, the product of activated neutrophils, on the development of collagen-induced arthritis in DBA 1/J mice; Kwasny-Krochin B et al.; Taurine chloramine (TauCl), a product of neutrophil myeloperoxidase - halide system, formed by a reaction of taurine with HOCl, is known as an anti-microbial and anti-inflammatory long-lived oxidant . We previously reported that TauCl inhibits in vitro the production of proinflammatory cytokines (IL-6, IL-8) by RA synoviocytes . Therefore we performed this study to investigate the effect of TauCl treatment on the development of collagen-induced arthritis (CIA) in DBA1/J mice . Early administration of TauCl (after primary immunization) resulted in the delay of the onset of CIA, but had no effect on severity of arthritis . TauCl, given daily for 21 days after booster immunization, did not reduce the symptoms of arthritis in those mice, which already developed CIA, but significantly diminished incidence of the disease (55% vs . 90% of placebo mice) . The mechanism of this effect is unknown . This is the first in vivo study suggesting that TauCl may be used for immune intervention in chronic inflammatory diseases. Crit Rev Biomed Eng, 2001, 29(5-6), 635 - 43 Experimental clinical study of the effect of millimeter waves on microbial and inflammatory renal diseases; Bagdasarova IV et al.; An investigation is made of the effect of low-intensity millimeter waves on the development of acute and chronic pyelonephritis . It is shown that the application of millimeter waves diminishes the rate of pyelonephritis recurrence and reduces the probability of chronic pyelonephritis . Furthermore, these waves normalize lipid peroxidation, stimulate antioxidant protection, and improve blood rheology . Experimental investigations performed on animals provided evidence for the positive effect of millimeter waves on the morphofunctional state of organs and tissues as well as on blood circulation . This especially concerns pyelonephritis of mycoplasmic etiology. Int J Pharm, 2002 Dec 5, 249(1-2), 23 - 31 Binders for colon specific drug delivery: an in vitro evaluation; Sinha VR et al.; The aim of the present study was to develop a single unit, site-specific drug formulation allowing targeted drug release in the colon . Tablets were prepared using polysaccharides or synthetic polymer as binders . These included xanthan gum, guar gum, chitosan and Eudragit E . Indomethacin was used as a model drug . The prepared tablets were enteric coated with Eudragit-L 100 to give protection in the stomach . The coated tablets were tested in-vitro for their suitability as colon specific drug delivery systems . The drug release studies were carried out in simulated stomach environment (pH 1.2) for 2 h followed by small intestinal environment at pH 6.8 . The dissolution data obtained from tablets demonstrates that the dissolution rate of the tablet is dependent upon the type and concentration of polysaccharide/polymer used as binder . The results demonstrate that enteric coated tablets containing 3% chitosan as a binder, showed only 12.5% drug release in the first 5 h, which is the usual upper gastrointestinal transit time, whereas, tablets prepared using guar gum as binder, were unable to protect drug release under similar conditions . Preparations with xanthan gum as a binder formed time-dependent release formulations . When used in a concentration of 5.92% in the tablets, 28% drug release was observed in the usual upper gastrointestinal tract conditions . It was also found that enteric coated preparation formulated with 8.88% of Eudragit E as binder could be used to carry water insoluble drug molecules to the colon especially in IBD . The above study shows that chitosan could be successfully used as a binder, for colon targeting of water insoluble drugs in preference to guar gum when used in the same concentration . Additionally, formulations developed with chitosan and Eudragit E would be highly site specific since drug release would be at a retarded rate till microbial degradation or polymer solubilization takes place in the colon. Environ Sci Technol, 2002 Nov 1, 36(21), 4649 - 55 Uptake and leaching of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine by hybrid poplar trees; Yoon JM et al.; The feasibility of remediating a high explosive, octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), using hybrid poplar trees (Populus deltoides x nigra, DN34) was investigated . The fate, transport, and toxicity were determined . HMX was taken up by poplar cuttings from hydroponic solutions in long-term experiments (65 days) without evidence of toxicity . HMX was not toxic to actively growing hybrid poplar cuttings, even under saturated conditions . The measured log Kow for HMX was 0.19, less than other explosives, TNT, and RDX . However, the calculated transpiration stream concentration factor (TSCF) and root concentration factor (RCF) for HMX from an uptake study using radiolabeled {U-14C}HMX were 0.21 +/- 0.07 and 5.55 +/- 1.78 mL/g, respectively, both of which were intermediate between the values for TNT and ROX in previous reports . A 70% uptake of {U-14C}HMX was translocated and accumulated in leaves, and no metabolites were observed during a 65-day exposure using radiochromatography of plant tissue extracts . Most of the accumulated HMX (57%) in dried (fallen) poplar leaves was leached by deionized water after 5 days . Bioaccumulation in poplar trees and resolublization of HMX from leaves would be of significant ecological concern, and phytoremediation may not be warranted as a treatment option unless other processes occur under field conditions that degrade HMX to innocuous end products (e.g., photolysis, hydrolysis, or microbial degradation). Environ Sci Technol, 2002 Nov 1, 36(21), 4540 - 6 Kinetics of 1,1,1-trichloroethane transformation by iron sulfide and a methanogenic consortium; Gander JW et al.; To evaluate the effect of potential interactions between methanogenic bacteria and iron sulfide minerals during transformation of 1,1,1-trichloroethane (1,1,1-TCA), we measured the kinetics of 1,1,1-TCA transformation by mackinawite (FeS(1 - x), but abbreviated as FeS) and a methanogenic consortium enriched on lactate (termed LEC) . Results from batch kinetic experiments show that 1,1,1-TCA transformation by FeS and resting LEC can be described by second-order rate expressions, with rates depending on 1,1,1-TCA concentration (M), FeS surface area concentration (m2 L(-1)), and LEC concentration (as measured by mg L(-1) volatile suspended solids (VSS)) . In reactors containing FeS alone, 1,1-dichloroethane (1,1-DCA) and 2-butyne were identified as products, but only accounted for 6% of the 1,1,1-TCA transformed . In reactors containing LEC alone, the only identified product was 1,1-DCA, which accounted for 46 +/- 8% of the 1,1,1-TCA transformed . Supernatant from LEC-alone reactors also transformed 1,1,1-TCA, suggesting that 1,1,1-TCA may be transformed by some non-cell component (such as a extracellular compound excreted by the organisms) that either reacts directly with 1,1,1-TCA or with the abiotic media to form a reactive species . Comparison of 1,1,1-TCA transformation rates from experiments with combinations of FeS (varying surface area concentrations) and LEC (varying VSS concentrations) to those with just FeS alone or LEC alone suggests some synergism occurs between the two reactive species . Observed enhancements took the form of faster 1,1,1-TCA transformation and faster 1,1-DCA appearance but less production of 1,1-DCA per unit of 1,1,1-TCA transformed . These observations suggest that the faster 1,1,1-TCA transformation in the combined systems (compared to the FeS-alone and LEC-alone experiments) is due to increased reactivity of both FeS and LEC, possibly due to production of soluble microbial products that make the FeS more reactive or less inhibition of LEC by 1,1,1-TCA due to FeS transformation of 1,1,1-TCA. Crit Rev Immunol, 2002, 22(2), 105 - 14 Regulation of macrophage-derived chemokine (MDC, CCL22) production; Yamashita U et al.; Macrophage-derived chemokine (MDC, CCL22) is a member of the CC-chemokine family and is composed of 69 amino acid residues . MDC is mainly produced by macrophages and dendritic cells upon the stimulation with microbial products, or anti-CD40 antibody, and is upregulated by TH2-type cytokines, such as IL-4 and -5, but is downregulated by TH1-type cytokines, such as IFN-gamma . MDC-production is also upregulated by prostaglandin and cyclic AMP-elevating agents . MDC causes chemotactic migration of dendritic cells and TH2 cells . Furthermore, MDC is highly-expressed in the lesions of TH2-related diseases, such as airway hypersensitivity and atopic dermatitis . Thus, MDC plays an important role in the recruitment of TH2 cells into the inflammatory sites and the regulation of TH2-related immune responses. Chemosphere, 2002 Nov, 49(6), 559 - 68 Influence of heavy metals on the microbial degradation of diesel fuel; Riis V et al.; The degradation of diesel fuel by a microbial community from a soil polluted by heavy metals (h.m.) in the presence of Cu, Ni, Zn, Pb, Cd, Hg and Cr (as chromate) was investigated . Experiments were conducted with soil slurries and the extracted community in liquid cultivation . The concentrations applied were in the sub-mM and mM range . Whereas the slurries displayed no significant effect, degradation in liquid culture was increasingly inhibited by higher metal concentrations . The course of degradation in suspension was demonstrated by the oxygen consumption . The order of toxicity was found to be: Hg > Cr(VI) > Cu > Cd > Ni > Pb > Zn . The absence of any effect for slurries was due to the non-availability of the metals in the soil, and to precipitation or adsorption to the soil in the case of amendment . The paper also includes results on the availability of h.m . and changes to the community after exposure. Ky Nurse . 2002 Oct-Dec;50(4):15. Impact of a 5-minute scrub on the microbial flora found on artificial, polished, or natural fingernails of operating room personnel; Clark JP; Edel et al.'s (1998) study gave support to the idea that fingernail treatments may put patients at increased risk due to the bacteria carried by nurses or other health care providers . The results of this study could be used to support a research utilization project to educate nurses of the increased risk of infecting patients due to fingernail treatments . Feasibility issues include the cost and time to revise hospital policies concerning fingernail treatments and to educate nurses in the workplace . Future research is needed with larger sample sizes and to investigate possible transmission of infectious bacteria to patients. Planta, 2002 Nov, 216(1), 155 - 61 Epub 2002 Nov 12. Occurrence and formation of indole-3-acetamide in Arabidopsis thaliana; Pollmann S et al.; An HPLC/GC-MS/MS technique (high-pressure liquid chromatography in combination with gas chromatography-tandem mass spectrometry) has been worked out to analyze indole-3-acetamide (IAM) with very high sensitivity, using isotopically labelled IAM as an internal standard . Using this technique, the occurrence of IAM in sterile-grown Arabidopsis thaliana (L.) Heynh . was demonstrated unequivocally . In comparison, plants grown under non-sterile conditions in soil in a greenhouse showed approximately 50% higher average levels of IAM, but the differences were not statistically significant . Thus, microbial contributions to the IAM extracted from the tissue are likely to be minor . Levels of IAM in sterile-grown seedlings were highest in imbibed seeds and then sharply declined during the first 24 h of germination and further during early seedling development to remain below 20-30 pmol g(-1) fresh weight throughout the rosette stage . The decline in indole-3-aetic acid (IAA) levels during germination was paralleled by a similar decline in IAM levels . Recombinant nitrilase isoforms 1, 2 and 3, known to synthesize IAA from indole-3-acetonitrile, were shown to produce significant amounts of IAM in vitro as a second end product of the reaction besides IAA . NIT2 was earlier shown to be highly expressed in developing and in mature A . thaliana embryos, and NIT3 is the dominantly active gene in the hypocotyl and the cotyledons of young, germinating seedlings . Collectively, these data suggest that the elevated levels of IAM in seeds and germinating seedlings result from nitrilase action on indole-3-acetonitrile, a metabolite produced in the plants presumably from glucobrassicin turnover. Genome Biol . 2002 Oct 25;3(11):research0064 . Epub 2002 Oct 25. The society of genes: networks of functional links between genes from comparative genomics; Yanai I et al.; BACKGROUND: Comparative genomics provides at least three methods beyond traditional sequence similarity for identifying functional links between genes: the examination of common phylogenetic distributions, the analysis of conserved proximity along the chromosomes of multiple genomes, and observations of fusions of genes into a multidomain gene in another organism . We have previously generated the links according to each of these methods individually for 43 known microbial genomes . Here we combine these results to construct networks of functional associations . RESULTS: We show that the functional networks obtained by applying these methods have different topologies and that the information they provide is largely additive . In particular, the combined networks of functional links contain an average of 57% of an organism's complete genetic complement, uncover substantial portions of known pathways, and suggest the function of previously unannotated genes . In addition, the combined networks are qualitatively different from the networks obtained using individual methods . They have a dominant cluster that contains approximately 80%-90% of the genes, independent of genome size, and the dominant clusters show the small world behavior expected of a biological system, with global connectivity that is nearly random, and local properties that are highly ordered . CONCLUSIONS: When the information on functional linkage provided by three emerging computational methods is combined, the integrated network uncovers large numbers of conserved pathways and identifies clusters of functionally related genes . It therefore shows considerable utility and promise as a tool for understanding genomic structure, and for guiding high throughput experimental investigations. Genome Biol . 2002 Oct 10;3(11):RESEARCH0060 . Epub 2002 Oct 10. Genomic functional annotation using co-evolution profiles of gene clusters; Zheng Y et al.; BACKGROUND: The current speed of sequencing already exceeds the capability of annotation, creating a potential bottleneck . A large proportion of the genes in microbial genomes remains uncharacterized . Here we propose a new method for functional annotation using the conservation patterns of gene clusters . If several gene clusters show the same coevolution pattern across different genomes it is reasonable to infer they are functionally related . The gene cluster phylogenetic profile integrates chromosomal proximity information and phylogenetic profile information and allows us to infer functional dependences between the gene clusters even at great distance on the chromosome . RESULTS: As a proof of concept, we applied our method to the genome of Escherichia coli K12 strain . Our method establishes functional relationships among 176 gene clusters, comprising 738 E . coli genes . The accuracy of pair phylogenetic profiles was compared with the single-gene phylogenetic profile and was shown to be higher . As a result, we are able to suggest functional roles for several previously unknown genes or unknown genomic regions in E . coli . We also examined the robustness of coevolution signals across a larger set of genomes and suggest a possible upper limit of accuracy for the phylogenetic profile methods . CONCLUSIONS: The higher-order phylogenetic profiles, such as the gene-pair phylogenetic profiles, can detect functional dependences that are missed by using conventional single-gene phylogenetic profile or the chromosomal proximity method only . We show that the gene-pair phylogenetic profile is more accurate than the single-gene phylogenetic profiles. J Agric Food Chem, 2002 Nov 20, 50(24), 6981 - 8 Peptides with angiotensin I-converting enzyme (ACE) inhibitory activity from defibrinated, hydrolyzed bovine plasma; Wanasundara PK et al.; Defibrinated bovine plasma (DBP) was treated with the microbial protease Flavourzyme to obtain protein hydrolysates with various degrees of hydrolysis (DH) . The angiotensin I-converting enzyme (ACE) inhibiting activity of the hydrolyzed protein was assessed with hippuryl-His-Leu as the substrate . The amount of hippuric acid released, due to uninhibited ACE activity, was determined by high-performance liquid chromatography . ACE inhibiting (ACEI) activity was found to increase with increasing DH; the 43% DH hydrolysate exhibited the highest activity and had an IC(50) of 1.08 mg/mL . Peptide fractions with high ACEI activity were isolated using size exclusion chromatography . The fraction that possessed the highest ACEI activity contained peptides with GYP, HL(I), HPY, HPGH, L(I)F, SPY, and YPH sequence motifs, as determined by reversed-phase liquid chromatography-tandem mass spectrometry using a novel immonium precursor-ion scanning technique . Some of these motifs correspond to sequences found in bovine serum albumin, a potential source of ACEI peptides in bovine plasma. Curr Rheumatol Rep, 2002 Dec, 4(6), 466 - 73 Heat shock proteins in juvenile idiopathic arthritis: keys for understanding remitting arthritis and candidate antigens for immune therapy; Prakken B et al.; Juvenile idiopathic arthritis (JIA) is in a majority of the cases of self-limiting, and sometimes even a self-remitting, disease . A growing amount of data suggests that active T cell regulation determines, at least partly, the clinical outcome of JIA . In experimental models of arthritis, a group of highly conserved microbial proteins, heat shock proteins (hsps), can be used to effectively prevent and treat arthritis . This protection is mediated through the induction of cross-reactive T cell responses to self-hsps . In JIA, naturally occurring T cell immune responses to hsps are associated with disease remission in restricted oligoarticular JIA . Moreover, those responses are associated with the induction of T cells with a regulatory phenotype . Taken together, these data imply that immune modulation with hsps can be an effective way to restore natural occurring T cell responses, and, thus, treat JIA and rheumatoid arthritis. J Biol Chem, 2003 Jan 17, 278(3), 1594 - 602 Epub 2002 Nov 07. Proteasomes modulate conjugation to the ubiquitin-like protein, ISG15; Liu M et al.; ISG15 is a ubiquitin-like protein that is induced by interferon and microbial challenge . Ubiquitin-like proteins are covalently conjugated to cellular proteins and may intersect the ubiquitin-proteasome system via common substrates or reciprocal regulation . To investigate the relationship between ISG15 conjugation and proteasome function, we treated interferon-induced cells with proteasome inhibitors . Surprisingly, inhibition of proteasomal, but not lysosomal, proteases dramatically enhanced the level of ISG15 conjugates . The stimulation of ISG15 conjugates occurred rapidly in the absence of protein synthesis and was most dramatic in the cytoskeletal protein fraction . Inhibition of ISG15 conjugation by ATP depletion abrogated the proteasome inhibitor-dependent increase in ISG15 conjugates, suggesting that the effect was mediated by de novo conjugation, rather than protection from proteasomal degradation or inhibition of ISG15 deconjugating activity . The increase in ISG15 conjugates did not occur through a stabilization of the ISG15 E1 enzyme, UBE1L . Furthermore, simultaneous modification of proteins by both ISG15 and ubiquitin did not account for the proteasome inhibitor-dependent increase in ISG15 conjugates . These findings provide the first evidence for a link between ISG15 conjugation and proteasome function and support a model in which proteins destined for ISG15 conjugation are proteasome-regulated. Gene, 2002 Oct 2, 298(2), 129 - 39 Molecular cloning of the fish interferon stimulated gene, 15 kDa (ISG15) orthologue: a ubiquitin-like gene induced by nephrotoxic damage; Liu M et al.; In mammals, the response to nephrotoxicant-induced renal injury is limited to repair of the proximal tubule by surviving epithelial cells . In contrast, bony fish are capable of both repair, and de novo production of nephrons in response to renal damage . Importantly, toxicant-induced nephron neogenesis in goldfish (Carassius auratus) parallels nephron development in the mammalian embryo, providing a vertebrate model for kidney development . We utilized this model system to identify genes induced by the renal toxicant, gentamicin, that may function in nephron neogenesis . A novel ubiquitin-like (UBL) gene, 40.1, was identified by differential display analysis of control and gentamicin-treated goldfish kidney . 40.1 was induced dramatically 3-7 days following a sublethal dose of gentamicin, and returned to basal level by 14 days post-treatment . The induction of 40.1 coincided with early renal injury in the proximal tubules of gentamicin-injected fish; however, expression was not restricted to the kidney, suggesting that 40.1 induction may be a more general response to cell injury . Sequence analysis revealed that 40.1 contains tandem UBL domains, and shares homology with ISG15, a 15 kD interferon-(IFN) stimulated UBL found in mammals . Analysis of the genome database for the pufferfish, Fugu rubrides, identified a goldfish ISG15 (gfISG15) homologue with an IFN-stimulated response element in the promoter region, providing further evidence that gfISG15 is the true teleost ISG15 orthologue . Zebrafish and catfish ISG15 genes were subsequently identified by sequence analysis . Consistent with its predicted function as a UBL, gfISG15 formed conjugates with cellular proteins in vitro and in transient transfections . Similar to the induction of mammalian ISG15 by microbial challenge, gfISG15 was induced in the spleen of mycobacteria-infected fish . These studies identified the first teleost ISG15 orthologue . The induction of gfISG15 as an early genetic event in response to a renal toxicant, and its conserved, stress-associated, expression in higher vertebrates suggests that ISG15 is an important component of the host response to diverse stress stimuli. Biomacromolecules, 2002 Nov-Dec, 3(6), 1327 - 35 Chemical modification of chlorinated microbial polyesters; Arkin AH et al.; Chlorination of microbial polyesters poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxyoctanoate) (PHO) was carried out by passing chlorine gas through their solutions . The chlorine contents in chlorinated PHB (PHB-Cl) and chlorinated PHO (PHO-Cl) were between 5.45 and 23.81 wt % and 28.09 and 39.09 wt %, respectively . Molecular weights of the chlorinated samples were in the range of between one-half to one-fourth of the original values because of hydrolysis during the chlorination process . Thermal properties of the PHO-Cl were dramatically changed with an increase in its glass transition (T(g) = 2 degrees C) and the melting transition (T(m)) . The T(g) of PHB-Cl varied from -20 to 10 degrees C, and its T(m) decreased to 148 degrees C . The chlorinated poly(3-hydroxyalkanoate)s (PHA-Cl) were converted to their corresponding quaternary ammonium salts (PHA-N(+)R(3)), sodium sulfate salts (PHA-S), and phenyl derivatives (PHA-Ph) . Cross-linked polymers were also formed by a Friedel-Crafts reaction between benzene and PHA-Cl . The modified PHO derivatives were characterized by (1)H NMR and (13)C NMR spectrometry, Fourier transform infrared spectroscopy, gel permeation chromatography, and differential scanning calorimetry techniques. J Biotechnol, 2003 Jan 23, 100(2), 161 - 8 Optimizing the formation of in vitro sponge primmorphs from the Chinese sponge Stylotella agminata (Ridley); Zhang W et al.; The establishment and optimization of in vitro primmorph formation from a Chinese sponge, Stylotella agminata (Ridley), collected from the South China Sea, were investigated . Our aims were to identify the key factors affecting primmorph formation in this species and to optimize the technique for developing an in vitro primmorph culture system . The size of dissociated cells from S . agminata is relatively small, in the range between 5 and 10 microm . Round-shaped primmorphs of less than 100 microm were formed 3 days after transferring the dissociated cells into seawater containing Ca(2+) and Mg(2+) . The effect of various cell dissociation conditions, inoculum cell density, concentration of antibiotics, pH, and temperature was further investigated upon the formation of primmorphs . The time required for primmorph formation, primmorph size distribution, and the proliferating capability were microscopically documented . Healthy sponge S . agminata, inoculum cell density and culture temperature play a critical role for the successful formation of primmorphs and that the microbial contamination will have to be controlled. J Biotechnol, 2003 Jan 23, 100(2), 119 - 25 Development in primary cell culture of demosponges; De Rosa S et al.; We have established primary cell culture of the marine demosponge Dysidea avara and Suberites domuncula . Microbial contamination was controlled by the use of a pool of antibiotics confirming the goodness of this procedure . Effect of pH, temperature and light was studied to establish the better growth conditions . The comparison of lipid composition of sponge and cells suggested a series of experiments to optimise the medium . A glucose dose-dependent experiment showed that the ideal glucose concentration is 1 g l(-1) . Supplementing the medium with unsaturated fatty acid and retinol, no promotion of growth was observed, but the compounds were totally metabolised by cells . Increments from 70 to 160% in the number of cells were observed, supplementing the medium with different concentration of cholesterol . These results suggest that the analysis of the chemical composition of sponge and cells give indication on the composition of the nutrient media. FEMS Microbiol Lett, 2002 Oct 29, 216(1), 1 - 7 Deprogrammed sporulation in Streptomyces; Ohnishi Y et al.; The bacterial genus Streptomyces forms chains of spores by septation at intervals in aerial hyphae and subsequent maturation on solid medium . Substrate hyphae undergo extensive lysis, liberating nutrients on which aerial hyphae develop . Some mutant strains, however, ectopically form spores by septation in substrate hyphae on solid medium or in vegetative hyphae in liquid medium, which suggests that all hyphae have the potential to differentiate into spores . A Streptomyces griseus mutant strain NP4, which has a mutation in the regulatory system for an ATP-binding cassette (ABC) transporter gene, forms ectopic spores in substrate hyphae only on glucose-containing medium . In addition, overexpression of a substrate-binding protein of the ABC transporter in the wild-type strain causes ectopic septation in very young substrate hyphae and subsequent sporulation in response to glucose . These ectopic spores germinate normally . The ectopic sporulation is independent of A-factor, a microbial hormone that determines the timing of aerial mycelium formation during normal development . Thus, substrate hyphae of Streptomyces have a potential to develop into spores without formation of aerial hyphae . For programmed development, therefore, the strict repression of septum formation in substrate mycelium should be necessary, as well as the positive signal relay leading to aerial mycelium formation followed by septation and sporulation. Australas J Dermatol, 2002 Nov, 43(4), 247 - 54 Atopic dermatitis: review of immunopathogenesis and advances in immunosuppressive therapy; Meagher LJ et al.; This paper reviews the theories of the pathogenesis of atopic dermatitis (AD), with a particular emphasis on its immunopathogenesis . The contribution of predisposing factors, immunopathogenic factors and provoking factors in the pathogenesis of AD are considered . Predisposing factors explored in this article include genetics and the disturbance of skin function . Immunopathogenic factors reviewed include T cell dysfunction, biphasic cytokine expression and the role of immunoglobulin E . Provoking factors considered include microbial factors, psychosomatic interactions, contact allergens and irritants, inhalant allergens, food and climate . Immunosuppressive treatments reviewed include cyclosporin, azathioprine, methotrexate, tacrolimus, interferon-gamma, phosphodiesterase inhibitors and pimecrolimus (SDZ ASM 981). J Matern Fetal Neonatal Med, 2002 Jul, 12(1), 35 - 41 Antibiotic administration in patients with preterm premature rupture of membranes reduces the rate of histological chorioamnionitis: a prospective, randomized, controlled study; Ovalle A et al.; OBJECTIVE: To determine whether antibiotic administration in patients with preterm premature rupture of membranes is associated with a reduction in the rate of histological chorioamnionitis and funisitis . METHODS: One hundred consecutive patients with preterm premature rupture of membranes and no labor between 24 and 34 weeks were invited to participate in this study . Eligible patients randomly received either clindamycin-gentamicin for 7 days or placebo, and were managed expectantly until 35 weeks unless fetal or maternal indications developed . Microbial invasion of the amniotic cavity was defined as the presence of a positive amniotic fluid culture obtained by transabdominal amniocentesis . Cervicovaginal infection was diagnosed when bacterial vaginosis or a positive culture for cervicovaginal pathogens or facultative bacteria associated with a significant increase in the white blood cell count were found . Histological chorioamnionitis was based on the observation of polymorphonuclear leukocyte infiltration of the chorionic plate or the extraplacental fetal membranes . Funisitis was diagnosed in the presence of polymorphonuclear leukocyte infiltration into the umbilical vessel walls or Wharton jelly . Statistics were performed using contingency tables . RESULTS: Seventy-one patients with available histological study of the placenta were included . Thirty-five women received antibiotics and 36 were given placebo . Patients who received antibiotics had a significantly lower rate of histological chorioamnionitis than patients who received placebo (46% (16/35) vs . 69% (25/36), respectively; p < 0.05) . This effect was more pronounced among women with microbial invasion of the amniotic cavity and/or cervicovaginal infection (58% vs . 89%, respectively; p < 0.01) . Antibiotic therapy was associated with an increase in the frequency of placentas without histological abnormalities (29% vs . 6%; p < 0.01) . The frequency of funisitis was not different between groups . CONCLUSION: Administration of antibiotics in patients with preterm premature rupture of membranes is associated with a significant reduction in the incidence of histological chorioamnionitis but it does not modify the frequency of funisitis. J Immunol, 2002 Nov 15, 169(10), 6030 - 5 DNA vaccination with heat shock protein 60 inhibits cyclophosphamide-accelerated diabetes; Quintana FJ et al.; Nonobese diabetic (NOD) mice spontaneously develop diabetes as a consequence of an autoimmune process that can be inhibited by immunotherapy with the 60-kDa heat shock protein (hsp60), with its mycobacterial counterpart 65-kDa (hsp65), or with other Ags such as insulin and glutamic acid decarboxylase (GAD) . Microbial infection and innate signaling via LPS or CpG motifs can also inhibit the spontaneous diabetogenic process . In addition to the spontaneous disease, however, NOD mice can develop a more robust cyclophosphamide-accelerated diabetes (CAD) . In this work, we studied the effect on CAD of DNA vaccination with constructs encoding the Ags human hsp60 (phsp60) or mycobacterial hsp65 (phsp65) . Vaccination with phsp60 protected NOD mice from CAD . In contrast, vaccination with phsp65, with an empty vector, or with a CpG-positive oligonucleotide was not effective, suggesting that the efficacy of the phsp60 construct might be based on regulatory hsp60 epitopes not shared with its mycobacterial counterpart, hsp65 . Vaccination with phsp60 modulated the T cell responses to hsp60 and also to the GAD and insulin autoantigens; T cell proliferative responses were significantly reduced, and the pattern of cytokine secretion to hsp60, GAD, and insulin showed an increase in IL-10 and IL-5 secretion and a decrease in IFN-gamma secretion, compatible with a shift from a Th1-like toward a Th2-like autoimmune response . Our results extend the role of specific hsp60 immunomodulation in the control of beta cell autoimmunity and demonstrate that immunoregulatory networks activated by specific phsp60 vaccination can spread to other Ags targeted during the progression of diabetes, like insulin and GAD. J Immunol, 2002 Nov 15, 169(10), 5827 - 37 A unique mechanism for innate cytokine promotion of T cell responses to viral infections; Pien GC et al.; The kinetics of CD8 T cell IFN-gamma responses as they occur in situ are defined here during lymphocytic choriomeningitis virus (LCMV) infections, and a unique mechanism for the innate cytokines IFN-alphabeta and IL-18 in promoting these responses is defined . Infections of mice with Armstrong or WE strains of LCMV induced an unexpectedly early day 4 IFN-gamma response detectable in serum samples and spleen and liver homogenates . Production of IFN-gamma was MHC class I/CD8 dependent, but did not require IL-12, NK cells, TCR-gammadelta T cells, MHC class II, or CD4 T cells . Peak response required specific Ag recognition, as administration of antagonist peptide partially impaired day 4 IFN-gamma induction, and viral peptide stimulation enhanced CD8 T cell IFN-gamma expression in culture . The IFN-gamma response was associated with IL-18 and IFN-alphabeta expression . Furthermore, both factors augmented peptide-driven IFN-gamma production in culture, and mice lacking IL-18 or IFN-alphabeta functions had reduced day 4 IFN-gamma . Collectively, these results demonstrate that during viral infections, there is a dramatic in vivo CD8 T cell response preceding maximal expansion of these cells, and that the mechanism supporting this response is dependent on endogenous innate cytokines . Because stimulation by microbial products is linked to innate cytokine expression, the studies also suggest a pathway for precisely limiting T cell functions to times of need. Bioresour Technol, 2003 Jan, 86(1), 95 - 8 Simplified method for estimation of microbial activity in compost by ATP analysis; Horiuchi JI et al.; A simplified method using ATP analysis was proposed to estimate the time course of microbial activity during composting . A compost sample was suspended in distilled water and its filtrate was used to estimate the ATP content in the compost by the luciferine-luciferase fluorescence reaction . The method permitted the rapid and simple determination of ATP in the compost and was successfully applied to monitor the time course of the microbial activity in the laboratory-scale composting process . The analytical simplicity in this method greatly improved the field-applicability of the ATP analysis for the composting process monitoring. Water Res, 2002 Oct, 36(17), 4193 - 202 Comparison of anaerobic dechlorinating enrichment cultures maintained on tetrachloroethene, trichloroethene, cis-dichloroethene and vinyl chloride; Duhamel M et al.; An anaerobic mixed microbial culture was enriched from soil and groundwater taken from a site contaminated with trichloroethene (TCE) . This enrichment culture was divided into four subcultures amended separately with either perchloroethene (PCE), TCE, cis-dichloroethene (cDCE) or vinyl chloride (VC) . In each of the four subcultures, the chlorinated ethenes were rapidly, consistently, and completely converted to ethene at rates of 30-50 micromol/l of culture per day, or an average 160 micro-electron equivalents/l of culture per day . These cultures were capable of sustained and rapid dechlorination of VC, and could not dechlorinate 1,2-dichloroethane, differentiating them from Dehalococcoides ethenogenes, the only known isolate capable of complete dechlorination of PCE to ethene . Chloroform (CF) and 1,1,1-trichloroethane, frequent groundwater co-contaminants with TCE and PCE, inhibited chlorinated ethene dechlorination . Most strongly inhibited was the final conversion of VC to ethene, with complete inhibition occurring at an aqueous CF concentration of 2.5 microM . Differences in rates and community composition developed between the different subcultures, including the loss of the VC enrichment culture's ability to dechlorinate PCE . Denaturing gradient gel electrophoresis of amplified bacterial 16S rRNA gene fragments identified three different DNA sequences in the enrichment cultures, all phylogenetically related to D . ethenogenes . Based on the PCR-DGGE results and substrate utilization patterns, it is apparent that significant mechanistic differences exist between each step of dechlorination from TCE to ethene, especially for the last important dechlorination step from VC to ethene. Farmaco, 2002 Oct, 57(10), 809 - 17 Synthesis and anti-microbial activity of isothiosemicarbazones and cyclic analogues; Maccioni E et al.; It is known that some derivatives of both thiourea and thiosemicarbazide exhibit potent anti-microbial activity . In order to investigate the effects on the biological properties of structural modifications of such structures, we have synthesised and studied some arylidenisothiosemicarbazones . In this paper we report on the synthesis and structure-activity relationships of some isothiosemicarbazones, where the arylidene group has been replaced with a cycloalkyl group and the sulfur atom has been either differently substituted or enclosed in a thiazole ring. Proc Natl Acad Sci U S A, 2002 Nov 12, 99(23), 14728 - 33 Epub 2002 Nov 04. Suppression of bone resorption by madindoline A, a novel nonpeptide antagonist to gp130; Hayashi M et al.; IL-6 is a multifunctional cytokine involved in regulation of differentiation, antibody production, and growth of certain types of tumor cells . Its excessive production plays a major role in pathogenesis of multiple myeloma and postmenopausal osteoporosis . In the course of a screening program aimed at IL-6 inhibitor from microbial products, we found madindoline A (MDL-A) and madindoline B, which have a fuloindoline structure with diketocyclopentene bound to the methyl group . MDL-A has no cytotoxic activities . It inhibited only activities of both IL-6 and IL-11 without affecting the IL-6-specific signal transduction cascade, JAK2/STAT3 . In a dose-dependent manner {(3)H}MDL-A binds to gp130, which is a signal transducing 130-kDa glycoprotein, but formation of the trimeric complex IL-6/IL-6 receptor/gp130 was not inhibited, suggesting that MDL-A suppresses dimerization of trimeric complexes . Not only did MDL-A markedly inhibit IL-6- and IL-11-induced osteoclastogenesis in vitro, but it also inhibited IL-6-stimulated serum amyloid A production and bone resorption in an experimental model of postmenopausal osteoporosis in vivo by a different mechanism from that of 17beta-estradiol . Here we show that MDL-A has a highly selective inhibitory effect on IL-6 and IL-11 activities by inhibiting a gp130 activity while suppressing bone loss in ovariectomized mice . MDL-A is anticipated as a lead compound for treatment of hormone-dependent postmenopausal osteoporosis, which has no serious side effects, and as a new mechanism of action, gp130 blocking. J Exp Med, 2002 Nov 4, 196(9), 1253 - 62 Parasite-induced lipoxin A4 is an endogenous regulator of IL-12 production and immunopathology in Toxoplasma gondii infection; Aliberti J et al.; The production of interleukin (IL)-12 is critical for the development of interferon (IFN)-gamma-dependent resistance to Toxoplasma gondii . Nevertheless, when this response is dysregulated, such as occurs in the absence of IL-10, the uncontrolled inflammation that results can have lethal consequences for the host . Recently, we demonstrated that lipoxin (LX)A(4), an eicosanoid mediator that depends on 5-lipoxygenase (LO) for its biosynthesis, exerts a regulatory role on dendritic cell IL-12 production triggered artificially by a T . gondii extract . We now formally establish the physiological relevance of this pathway in the systemic control of IL-12 production induced by live T . gondii infection and demonstrate its function to be distinct from that of IL-10 . Thus, T . gondii-exposed wild-type, but not 5-LO-deficient animals, produced high levels of serum LXA(4) beginning at the onset of chronic infection . Moreover, 5-LO(-/-), in contrast to wild-type mice, succumbed during the same period displaying a marked encephalitis . The increased mortality of the 5-LO(-/-) animals was also associated with significant elevations of IL-12 and IFN-gamma and was completely prevented by the administration of a stable LXA(4) analogue . Together, these findings demonstrate a new pathway involving the induction of host LXs for the in vivo regulation of proinflammatory responses during microbial infection. J Neuroimmunol, 2002 Nov, 132(1-2), 41 - 8 Resistance to ocular herpes simplex virus type 1 infection in IL-12 transgenic mice; Al-Khatib K et al.; Interleukin-12 (IL-12) is a potent inflammatory cytokine that influences the innate and adaptive immune response to microbial pathogens including viruses . It was reasoned that constitutive IL-12 production in mice would enhance resistance to herpes simplex virus type 1 (HSV-1) infection . To test this hypothesis, transgenic mice expressing the p35 and p40 genes of IL-12 under a glial fibrillary acidic protein (GFAP) promoter were ocularly infected with HSV-1 . These mice displayed increased survival and reduced viral titers in the eye, trigeminal ganglion (TG), and brain stem in comparison to wild type controls . Consistent with these results, HSV-1 immediate early and early gene expression were reduced to 50-130-fold in the trigeminal ganglion of infected transgenic mice compared to infected, non-transgenic counterparts as determined by real time PCR . Associated with viral resistance, IL-12 and IFN-gamma mRNA levels and IL-12 protein were elevated in the eyes of the transgenic versus non-transgenic mice during the acute infection . Collectively, the data show the inherent resistance of mice constitutively expressing IL-12 to ocular HSV-1 infection-an outcome that is independent of the adaptive immune system at the time of infection. Intensive Care Med, 2002 Nov, 28(11), 1613 - 8 Epub 2002 Sep 21. Soluble L-selectin levels predict survival in sepsis; Seidelin JB et al.; OBJECTIVE: To evaluate serum soluble L-selectin as a prognostic factor for survival in patients with sepsis . DESIGN: A prospective study of mortality in patients with sepsis whose serum levels of sL-selectin were measured on admission to an intensive care unit (ICU) and 4 days later . Follow-up data on mortality were obtained from the Danish Central Office of Civil Registration . SETTING: A tertiary referral university hospital ICU in Copenhagen . PATIENTS: Sixty-three patients meeting the criteria for systemic inflammatory response syndrome (SIRS) with a suspected or verified infection in one or more major organs, and 14 control subjects . MEASUREMENTS AND RESULTS: On admission to the ICU the Simplified Acute Physiology Score (SAPS) II was calculated, and relevant microbial cultures were performed . Mortality was registered at various follow-up points: 7 days after admission, at discharge from hospital, and 3 and 12 months after admission . Serum sL-selectin levels were significantly lower in the patients than in the controls . Sepsis nonsurvivors had significantly lower levels than survivors . Efficiency analysis and receiver operation characteristics showed that the ideal cutoff point for sL-selectin as a test for sepsis survival was 470 ng/ml . The accumulated mortality in patients with subnormal sL-selectin levels on admission was significantly increased . No correlation was found between clinical or paraclinical markers, including SAPS II and sL-selectin, and no relationship to the microbial diagnosis was found . CONCLUSIONS: Serum sL-selectin is a predictor of survival in patients with sepsis . Those admitted with low sL-selectin (<470 ng/ml) are characterized by a high mortality within the subsequent 12-month period. Microb Ecol, 2003 Jan, 45(1), 20 - 8 Epub 2002 Nov 06. A two-species test of the hypothesis that spatial isolation influences microbial diversity in soil; Treves DS et al.; The hypothesis that spatial isolation is a key determinant of microbial community structure in soils was evaluated by examining the competitive dynamics of two species growing on a single resource in a uniform sand matrix under varied moisture content . One species dominated the community under highly connected, saturated treatments, suggesting that these conditions allow competitive interactions to structure the community . As moisture content decreased, however, the less competitive species became established in the community . This effect was most pronounced at a matric water potential of -0.14 MPa where estimates of final population density and species fitness were equal . A second but more closely related species pair exhibited a similar response to decreasing moisture, suggesting that the effects of spatial isolation we observed are not simply a species-pair-specific phenomenon . These findings indicate that spatial isolation, created by low moisture content, plays an important role in structuring soil microbial communities. Nat Immunol, 2002 Dec, 3(12), 1163 - 8 Epub 2002 Nov 04. CD1-dependent dendritic cell instruction; Vincent MS et al.; Both microbial products and T cell factors influence dendritic cell (DC) maturation . However, it is not known which T cells are capable of interacting with DCs at the initiation of adaptive immunity, when foreign antigen-specific T cells are rare . We show here that self-reactive CD1-restricted T cells can promote DC maturation by recognizing CD1 in the absence of foreign antigens . T cell recognition of all four CD1 isoforms can trigger DC maturation, but their distinct mechanisms of costimulation lead to profound differences in concomitant interleukin 12 p70 production . Distinct CD1-reactive T cells may thus differentially direct DC development early in the immune response, thereby controlling subsequent polarization of acquired immunity. J Anim Sci, 2002 Oct, 80(10), 2704 - 16 Apparent ruminal degradation and rumen escape of soluble nitrogen fractions in grass and grass silage administered intraruminally to lactating dairy cows; Volden H et al.; The main objective of this study was to investigate in vivo ruminal degradation and rumen escape of soluble N fractions in grass and grass silage . Soluble protein and long-chain peptides (PLP), small peptides (SP) and free AA (FAA) were obtained from fresh grass and grass silages fertilized with different levels of N . Soluble extracts from the forages were pulse dosed into the rumen of three cannulated lactating dairy cows, and a simple or complex model was used to examine the kinetics of the soluble N fractions in the rumen . When soluble extracts from silage were investigated, pulse dosages of total nonammonia N (NAN) were 21, 27, and 32 g, while for fresh grass only dosages of 20 g were ruminally administered . In the silage extracts, mean proportions of PLP-N, SP-N, and FAA-N in the NAN were 30, 52, and 18%, respectively, whereas in the fresh grass the corresponding values were 67, 20, and 13% . From silage extracts, all three soluble N fractions showed a linear decrease (P < 0.05) in degradation rate and an increase (P < 0.05) in ruminal escape with increasing dosage . In silage, mean degradation rates, parameterized from the complex model, were 230, 214, and 334%/h for PLP-N, SP-N, and FAA-N, respectively, and the ruminal escape was highest (P < 0.05) for SP-N (11.2% of dose) and lowest (P < 0.05) for FAA-N (5.0% of dose) . No differences in degradation rate and ruminal escape between fresh grass and silage were observed . However, the proportion of N dose converted to ammonia was only 24% in the fresh grass, whereas for the silages a mean value of 76% was found . From this study, it is concluded that a significant amount of dietary soluble N escapes ruminal degradation, and thus contributes to the intestinal AA supply . Moreover, if the main aim is to study degradation kinetics of individual N fractions, a complex model should be used in the evaluation . This model can also be used to study ruminal synchronization of N and energy for microbial growth. Clin Exp Immunol, 1989 Nov, 78(2), 189 - 95 Immunohistology of oral lesions from patients with recurrent oral ulcers and Behçet's syndrome; Poulter LW et al.; A qualitative and quantitative immunohistological investigation was performed on biopsies of oral ulcers from patients with Behcet's syndrome (BS) and those with recurrent oral ulcers (ROU) . The results were compared with control oral biopsies from patients with other diseases and normal oral mucosa . The expression of HLA-DR on the cell membrane of keratinocytes was found in 13 out of 15 lesions from patients with BS and ROU, as compared with only one out of 15 controls . The relative density of HLA-DR was investigated quantitatively by microdensitometry and this confirmed that DR expression in the epithelial cells of patients with BS and ROU was significantly greater than in diseased and normal control oral tissues . A prominent mononuclear cell infiltration consisted predominantly of T lymphocytes and mature macrophages . Analysis of the CD4 and CD8 subsets of T cells failed to show significant differences between BS, ROU and control diseased tissues . Increased numbers of Langerhans cells were found in the epithelium by morphometric analysis with the CD1 monoclonal antibody in BS and ROU but an increased number was also found in lichen planus . The results suggest that the immunohistological changes in oral lesions of BS and ROU manifest an enhanced immune response in the epithelium, keratinocytes express HLA-class II antigen and increased number of Langerhans cells as well as in the lamina propria with a prominent infiltration of CD4, CD8 and macrophage-like cells . The characteristic pattern of exacerbations and remissions of oral ulceration can be interpreted by the hypothesis that an initiating microbial agent may induce a mononuclear cell infiltration, with the release of cytokines, expression of class II antigen in keratinocytes and causing ulceration, followed by down-regulation of immunity by tolerant T cells induced by the class II positive keratinocytes, leading to a remission. Plant J, 2002 Nov, 32(3), 375 - 90 NPP1, a Phytophthora-associated trigger of plant defense in parsley and Arabidopsis; Fellbrich G et al.; Activation of non-cultivar-specific plant defense against attempted microbial infection is mediated through the recognition of pathogen-derived elicitors . Previously, we have identified a peptide fragment (Pep-13) within a 42-kDa cell wall transglutaminase from various Phytophthora species that triggers a multifacetted defense response in parsley cells . Many of these oomycete species have now been shown to possess another cell wall protein (24 kDa), that evoked the same pattern of responses in parsley as Pep-13 . Unlike Pep-13, necrosis-inducing Phytophthora protein 1 (NPP1) purified from P . parasitica also induced hypersensitive cell death-like lesions in parsley . NPP1 structural homologs were found in oomycetes, fungi, and bacteria, but not in plants . Structure-activity relationship studies revealed the intact protein as well as two cysteine residues to be essential for elicitor activity . NPP1-mediated activation of pathogen defense in parsley does not employ the Pep-13 receptor . However, early induced cellular responses implicated in elicitor signal transmission (increased levels of cytoplasmic calcium, production of reactive oxygen species, MAP kinase activation) were stimulated by either elicitor, suggesting the existence of converging signaling pathways in parsley . Infiltration of NPP1 into leaves of Arabidopsis thaliana Col-0 plants resulted in transcript accumulation of pathogenesis-related (PR) genes, production of ROS and ethylene, callose apposition, and HR-like cell death . NPP1-mediated induction of the PR1 gene is salicylic acid-dependent, and, unlike the P . syringae pv . tomato DC3000(avrRpm1)-induced PR1 gene expression, requires both functional NDR1 and PAD4 . In summary, Arabidopsis plants infiltrated with NPP1 constitute an experimental system that is amenable to forward genetic approaches aiming at the dissection of signaling pathways implicated in the activation of non-cultivar-specific plant defense. Scand J Immunol, 2002 Nov, 56(5), 518 - 21 Circulating, interferon-producing plasmacytoid dendritic cells decline during human ageing; Shodell M et al.; Increased frequency and severity of infections in the elderly have been taken as indicative of declining immune function . Dendritic cells (DCs), the most important antigen-presenting cells, play a central role in initiating and modulating immune responses . One type, DC2, arises from precursor plasmacytoid DCs (pDCs), a rare population of circulating blood cells, whose hallmark function is rapid and copious production of interferon-alpha (IFN-alpha) upon microbial challenge . We found significant decreases of the circulating pDCs during ageing in healthy adult humans, as defined both by flow cytometry and IFN-alpha generation . Mean pDC/mm3 in peripheral blood declined from 7.8 for the youngest age group (18-39 years) to 4.2 for the oldest (60-91 years; P = 0.017) . IFN-alpha generation declined similarly, from 3537 to 1201 IU/ml, respectively (P = 0.006) . There was also a slight decline over the age range in the amount of IFN generated per pDC (slope = -0.0087; P = 0.046) . CD4+ T cells decreased by approximately 20% over the same age range (P = 0.001), while there was no change in the total lymphocyte or monocyte counts. J Clin Microbiol, 2002 Nov, 40(11), 4244 - 50 Development of a peptide-mediated capture PCR for detection of Mycobacterium avium subsp . paratuberculosis in milk; Stratmann J et al.; Based on phage display technology, a peptide-mediated magnetic separation technique was developed to facilitate selective isolation of Mycobacterium avium subsp . paratuberculosis (M . paratuberculosis) from bulk milk of naturally infected dairy herds . Nine recombinant bacteriophages binding to M . paratuberculosis were isolated from a commercial phage-peptide library encoding random 12-mer peptides . Nucleotide sequencing revealed the deduced sequence of the binding peptides . One peptide with the sequence NYVIHDVPRHPA, designated aMP3, was chemically synthesized with an amino-terminal biotin residue attached via an amino-hexacarbonic acid spacer molecule . Paramagnetic beads coated with the phage or with peptide aMP3 enabled the capture of M . paratuberculosis from milk . Combining this peptide-mediated magnetic separation with an ISMav2-based PCR allowed the detection of M . paratuberculosis in artificially spiked milk down to a concentration of 10(1) ml(-1) . Experiments using milk from naturally infected cows and bulk milk samples from infected herds demonstrated that the peptide-media