Appl Environ Microbiol, 2001 Dec, 67(12), 5474 - 81 Proteins induced during adaptation of Acetobacter aceti to high acetate concentrations; Steiner P et al.; As a typical product of microbial metabolism, the weak acid acetate is well known for its cytotoxic effects . In contrast to most other microbes, the so-called acetic acid bacteria can acquire significant resistance to high acetate concentrations when properly adapted to such hostile conditions . To characterize the molecular events that are associated with this adaptation, we analyzed global protein expression levels during adaptation of Acetobacter aceti by two-dimensional gel electrophoresis . Adaptation was achieved by using serial batch and continuous cultivations with increasing acetate supplementation . Computer-aided analysis revealed a complex proteome response with at least 50 proteins that are specifically induced by adaptation to acetate but not by other stress conditions, such as heat or oxidative or osmotic stress . Of these proteins, 19 were significantly induced in serial batch and continuous cultures and were thus noted as acetate adaptation proteins (Aaps) . Here we present first microsequence information on such Aaps from A . aceti . Membrane-associated processes appear to be of major importance for adaptation, because some of the Aap bear N-terminal sequence homology to membrane proteins and 11 of about 40 resolved proteins from membrane protein-enriched fractions are significantly induced. Br J Haematol, 2001 Oct, 115(1), 186 - 94 Differentiation and expansion of endothelial cells from human bone marrow CD133(+) cells; Quirici N et al.; We report a method of purifying, characterizing and expanding endothelial cells (ECs) derived from CD133(+) bone marrow cells, a subset of CD34(+) haematopoietic progenitors . Isolated using immunomagnetic sorting (mean purity 90 +/- 5%), the CD133(+) bone marrow cells were grown on fibronectin-coated flasks in M199 medium supplemented with fetal bovine serum (FBS), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and insulin growth factor (IGF-1) . The CD133(+) fraction contained 95 +/- 4% CD34(+) cells, 3 +/- 2% cells expressing VEGF receptor (VEGFR-2/KDR), but did not express von Willebrand factor (VWF), VE-cadherin, P1H12 or TE-7 . After 3 weeks of culture, the cells formed a monolayer with a typical EC morphology and expanded 11 +/- 5 times . The cells were further purified using Ulex europaeus agglutinin-1 (UEA-1)-fluorescein isothiocyanate (FITC) and anti-FITC microbeads, and expanded with VEGF for a further 3 weeks . All of the cells were CD45(-) and CD14(-), and expressed several endothelial markers (UEA-1, VWF, P1H12, CD105, E-selectin, VCAM-1 and VE-cadherin) and typical Weibel-Palade bodies . They had a high proliferative potential (up to a 2400-fold increase in cell number after 3 weeks of culture) and the capacity to modulate cell surface antigens upon stimulation with inflammatory cytokines . Purified ECs were also co-cultivated with CD34(+) cells, in parallel with a purified fibroblastic cell monolayer . CD34(+) cells (10 x 10(5)) gave rise to 17,951 +/- 2422 CFU-GM colonies when grown on endothelial cells, and to 12,928 +/- 4415 CFU-GM colonies on fibroblast monolayers . The ECs also supported erythroid blast-forming unit (BFU-E) colonies better . These results suggest that bone marrow CD133(+) progenitor cells can give rise to highly purified ECs, which have a high proliferative capacity, can be activated by inflammatory cytokines and are superior to fibroblasts in supporting haematopoiesis . Our data support the hypothesis that endothelial cell progenitors are present in adult bone marrow and may contribute to neo-angiogenesis. Planta, 2001 Oct, 213(6), 977 - 80 X-ray absorption spectroscopy study shows that the rapid selenium volatilizer, pickleweed (Salicornia bigelovii Torr.), reduces selenate to organic forms without the aid of microbes; Lee A et al.; In many plant species, selenium (Se) volatilization is limited by the reduction of selenate and its chemical conversion to organic Se compounds, a process that may be facilitated by rhizosphere microbes . This study was conducted to determine if pickleweed (Salicornia bigelovii Torr.), which is characterized by having high rates of Se volatilization from selenate, is able to reduce selenate into organic forms of Se axenically, or whether it requires the presence of microbes . X-ray absorption spectroscopy analysis showed that shoots and roots of pickleweed plants supplied with 50 microM selenate accumulated Se predominantly in organic Se forms (about 65-75% of the total accumulated Se), regardless of whether the plants were grown axenically or in the presence of microbes . The results suggest that, unlike other species for which selenate reduction appears to be rate limiting . e.g . Indian mustard (Brassica juncea L.) and broccoli (Brassica oleracea L.), pickleweed is unusual in that it has an enhanced capacity to reduce selenate to organic forms that is independent of the presence of rhizosphere microbes. Curr Biol, 2001 Nov 13, 11(22), R929 - 31 Evolution: constantly avoiding mutation; Sniegowski P; The genomic mutation rate of the archaeon Sulfolobus acidocaldarius, which inhabits a harsh and potentially mutagenic environment, surprisingly agrees well with the previously observed constancy of genomic mutation rates in microbes . The evolutionary explanation for this constancy of genomic mutation rates remains obscure. J Appl Physiol, 2001 Dec, 91(6), 2720 - 9 On the likelihood of decompression sickness during H(2) biochemical decompression in pigs; Fahlman A et al.; A probabilistic model was used to predict decompression sickness (DCS) outcome in pigs during exposures to hyperbaric H(2) to quantify the effects of H(2) biochemical decompression, a process in which metabolism of H(2) by intestinal microbes facilitates decompression . The data set included 109 exposures to 22-26 atm, ca . 88% H(2), 9% He, 2% O(2), 1% N(2), for 0.5-24 h . Single exponential kinetics described the tissue partial pressures (Ptis) of H(2) and He at time t: Ptis = integral (Pamb - Ptis) . tau(-1) dt, where Pamb is ambient pressure and tau is a time constant . The probability of DCS {P(DCS)} was predicted from the risk function: P(DCS) = 1 - e(-r), where r = integral (Ptis(H(2)) + Ptis(He) - Thr - Pamb) . Pamb(-1) dt, and Thr is a threshold parameter . Inclusion of a parameter (A) to estimate the effect of H(2) metabolism on P(DCS): Ptis(H(2)) = integral (Pamb - A - Ptis(H(2))) . tau(-1) dt, significantly improved the prediction of P(DCS) . Thus lower P(DCS) was predicted by microbial H(2) metabolism during H(2) biochemical decompression. J Appl Physiol, 2001 Dec, 91(6), 2713 - 9 Increasing activity of H(2)-metabolizing microbes lowers decompression sickness risk in pigs during H(2) dives; Kayar SR et al.; The risk of decompression sickness (DCS) was modulated by varying the biochemical activity used to eliminate some of the hydrogen (H(2)) stored in the tissues of pigs (19.4 +/- 0.2 kg) during hyperbaric exposures to H(2) . Treated pigs (n = 16) received intestinal injections of Methanobrevibacter smithii, a microbe that metabolizes H(2) to water and CH(4) . Surgical controls (n = 10) received intestinal injections of saline, and an additional control group (n = 10) was untreated . Pigs were placed in a chamber and compressed to 24 atm abs (20.6-22.9 atm H(2)) . After 3 h, the pigs were decompressed and observed for symptoms of DCS for 1 h . Pigs with M . smithii had a significantly lower (P < 0.05) incidence of DCS (44%; 7/16) than all controls (80%; 16/20) . The DCS risk decreased with increasing activity of microbes injected (logistic regression, P < 0.05) . Thus the supplemental tissue washout of the diluent gas by microbial metabolism was inversely correlated with DCS risk in a dose-dependent manner in this pig model. Sci Total Environ, 2001 Nov 12, 279(1-3), 45 - 50 Effects of elevated CO2 on fen peat biogeochemistry; Kang H et al.; Effects of elevated atmospheric CO2 concentration on northern peatland biogeochemistry was studied in a short-term experiment . Eight intact soil cores (11-cm diameter x 40-cm depth) with Juncus and Festuca spp . were collected from a calcareous fen in north Wales . Half of the cores were incubated under 350 ppm CO2 concentration, whilst the other four cores were maintained at 700 ppm CO2 . After a 4-month incubation, significantly higher biomass (root + shoot + algal mat) was determined under elevated CO2 conditions . Higher emissions of N2O and CO2, and higher concentration of pore-water DOC (dissolved organic carbon) were also observed under elevated CO2 . However, no significant differences were found in CH4 emission or soil enzyme activities (beta-glucosidase, phosphatase, and N-acetylglucosaminidase) in the bulk soil . Overall, the results suggest that elevated CO2 would increase the primary productivity of the fen vegetation, and stimulate N2O and CO2 emissions as a consequence of an enhanced DOC supply from the vegetation to the soil microbes. Biochem Soc Trans, 2001 Nov, 29(Pt 6), 853 - 9 Sepsis begins at the interface of pathogen and host; Beutler B; To the modern mind, the term 'sepsis' conjures up images of microbes . It is easy to forget that the word predates any understanding of the microbial origins of infectious disease . Derived from the Greek 'sepsios' (rotten), sepsis denotes decay: a phenomenon that humans once regarded as a mysterious though inevitable natural process . A living organism does not accept decay passively . Virtually all multicellular life forms are capable of resisting infection through the generation of a vigorous immune response . In mammals, the response is so stereotypic that it has come to define sepsis itself: it is often called the 'septic syndrome' . Our current understanding of the innate immune system is deeply rooted in the study of sepsis . The chain of events linking infection to tissue injury and cardiovascular collapse is not obvious, and affirmation of the concept required three major discoveries . First, the septic syndrome was found to be caused by toxic products of microbes . Secondly, these toxic substances were found to be toxic because of their propensity to activate cells of the innate immune system, prompting cytokine production . Thirdly, the activating events initiated by microbial toxins were traced to members of an ancient family of defensive molecules, versions of which operate in virtually all multicellular life forms . In mammals, proteins of this family are now known as Toll-like receptors . They represent a point of direct contact, and first contact, between a pathogen and the host immune system. Ground Water, 2001 Nov-Dec, 39(6), 886 - 94 Viral transport in a sand and gravel aquifer under field pumping conditions; Woessner WW et al.; Ground water supplies contaminated with microbes cause more than 50% of the water-borne disease outbreaks in the United States . Proposed regulations suggest natural disinfection as a possible mechanism to treat microbe-impacted ground water under favorable conditions . However, the usefulness of current models employed to predict viral transport and natural attenuation rates is limited by the absence of field scale calibration data . At a remote floodplain aquifer in western Montana, the bacteriophages MS2, phiX174, and PRD1; attenuated poliovirus type-1 (CHAT strain); and bromide were seeded as a slug 21.5 m from a well pumping at a steady rate of 408 L/min . Over the 47-hour duration of the test, resulting in the exchange of 12 to 13 pore volumes, 77% of the bromide, 55% of the PRD1, 17% of the MS2, 7% of the phiX174, and 0.12% of the poliovirus masses were recovered at the pumping well . Virus transport behavior was controlled by mechanical dispersion, preferential flow, time-dependent nonreversible and reversible attachment, and apparent mass transfer to immobile domains within the sand and gravel dominated aquifer . The percentage of virus recovery appears correlated with reported viral isoelectric point (pI) values . Successful modeling of viral transport in coarse-grained aquifers will require separation of viral specific properties from reported lumped viral-transport system parameters. Proc Natl Acad Sci U S A, 2001 Nov 20, 98(24), 13519 - 24 Epub 2001 Nov 13. The 2.0-A crystal structure of tachylectin 5A provides evidence for the common origin of the innate immunity and the blood coagulation systems; Kairies N et al.; Because invertebrates lack an adaptive immune system, they had to evolve effective intrinsic defense strategies against a variety of microbial pathogens . This ancient form of host defense, the innate immunity, is present in all multicellular organisms including humans . The innate immune system of the Japanese horseshoe crab Tachypleus tridentatus, serving as a model organism, includes a hemolymph coagulation system, which participates both in defense against microbes and in hemostasis . Early work on the evolution of vertebrate fibrinogen suggested a common origin of the arthropod hemolymph coagulation and the vertebrate blood coagulation systems . However, this conjecture could not be verified by comparing the structures of coagulogen, the clotting protein of the horseshoe crab, and of mammalian fibrinogen . Here we report the crystal structure of tachylectin 5A (TL5A), a nonself-recognizing lectin from the hemolymph plasma of T . tridentatus . TL5A shares not only a common fold but also related functional sites with the gamma fragment of mammalian fibrinogen . Our observations provide the first structural evidence of a common ancestor for the innate immunity and the blood coagulation systems. Vet Parasitol, 2001 Nov 22, 101(3-4), 261 - 74 Molecular technology and antigenic variation among intraerythrocytic hemoparasites: do we see reality? Allred DR. Antigenic variation is one mechanism of immune evasion utilized by many microorganisms--encompassing such broad evolutionary groups as viruses, bacteria, and protozoa--to survive the onslaught of a specifically activated host immune system . Because of its importance to the survival of many infectious agents there is considerable interest in understanding this phenomenon . With knowledge of the molecular mechanisms by which these microbes deliberately manipulate their genomes, it may be possible to disrupt the molecular machinery of the responsible genetic mechanisms . Among intraerythrocytic parasites, genetic mechanisms that have been observed or postulated to control antigenic variation include segmental gene conversion, epigenetically controlled in situ transcriptional switching, alterations of chromosomal structure associated with transcriptional control, and recombination during sexual reproduction . Likely, more than one type of mechanism is used by all organisms that undergo antigenic variation . In this paper, both the observed mechanisms and some of the molecular technology used to detect these mechanisms are discussed . While often seemingly straightforward from a technical standpoint, sometimes subtle differences in the methods used to study this process may affect what is observed . Some examples of this phenomenon are discussed in the context of a small selection of intraerythrocytic parasites. BMC Bioinformatics . 2001;2(1):9 . Epub 2001 Oct 16. FastGroup: a program to dereplicate libraries of 16S rDNA sequences; Seguritan V et al.; BACKGROUND: Ribosomal 16S DNA sequences are an essential tool for identifying and classifying microbes . High-throughput DNA sequencing now makes it economically possible to produce very large datasets of 16S rDNA sequences in short time periods, necessitating new computer tools for analyses . Here we describe FastGroup, a Java program designed to dereplicate libraries of 16S rDNA sequences . By dereplication we mean to: 1) compare all the sequences in a data set to each other, 2) group similar sequences together, and 3) output a representative sequence from each group . In this way, duplicate sequences are removed from a library . RESULTS: FastGroup was tested using a library of single-pass, bacterial 16S rDNA sequences cloned from coral-associated bacteria . We found that the optimal strategy for dereplicating these sequences was to: 1) trim ambiguous bases from the 5' end of the sequences and all sequence 3' of the conserved Bact517 site, 2) match the sequences from the 3' end, and 3) group sequences > or =97% identical to each other . CONCLUSIONS: The FastGroup program simplifies the dereplication of 16S rDNA sequence libraries and prepares the raw sequences for subsequent analyses. Biochemistry, 2001 Nov 20, 40(46), 13753 - 9 Visual detection of specific, native interactions between soluble and microbead-tethered alpha-helices from membrane proteins; Ashish et al.; Using peptides tethered to polymer microbeads, we have developed a technique for measuring the interactions between the transmembrane alpha-helices of membrane proteins and for screening combinatorial libraries of peptides for members that interact with specific helices from membrane proteins . The method was developed using the well-characterized homodimerization sequence of the membrane-spanning alpha-helix from the erythrocyte membrane protein glycophorin A (GPA) . As a control, we also tested a variant with a dimer-disrupting alteration of a critical glycine residue to leucine . To test for detectable, native interactions between detergent-solubilized and microbead-tethered alpha-helices, we incubated fluorescent dye-labeled GPA analogues in sodium dodecyl sulfate solution with microbeads that contained covalently attached GPA analogues . When the dye-labeled peptide in solution and the bead-tethered peptide both contained the native glycophorin A sequence, the microbeads readily accumulated the dye through lateral peptide-peptide interactions and were visibly fluorescent under UV light . When either the peptide in solution or the peptide attached to the beads contained the glycine to leucine change, the beads did not accumulate any dye . The usefulness of this method for screening tethered peptide libraries was tested by incubating dye-labeled, native sequence peptides in detergent solution with a few native sequence beads plus an excess of beads containing the variant glycine to leucine sequence . When the dye-labeled peptide in solution was present at a concentration of > or =2 microM, the few native sequence beads were visually distinguishable from the others because of their bright fluorescence . Using this model system, we have shown that it is possible to visually detect specific, native interactions between alpha-helices from membrane proteins using peptides tethered to polymer microbeads . It will thus be possible to use this method to measure the specific lateral interactions that drive the folding and organization of membrane proteins and to screen combinatorial libraries of peptides for members that interact with them. Acta Obstet Gynecol Scand, 2001 Nov, 80(11), 1036 - 8 Is preeclampsia an infectious disease? Trogstad LI, Eskild A, Bruu AL, Jeansson S, Jenum PA. BACKGROUND: Studies have suggested a strong paternal factor in the etiology of preeclampsia . If preeclampsia is caused by an infectious agent transmitted by the woman's partner, seronegative women who may experience primary infection in pregnancy should be at increased risk of preeclampsia as compared to previously infected women . The aim of this study was to assess the impact of being seronegative for some viruses transmitted by close contact on the risk of developing preeclampsia . METHODS: Nine hundred and seventy-eight women were randomly drawn from a basic study population of 35,940 pregnant women in Norway . A serum sample drawn at the first antenatal visit was analyzed for specific IgG antibodies against herpes simplex virus type-2, cytomegalovirus and Epstein-Barr virus . For comparison, antibody status against Toxoplasma gondii was also assessed . Information on preeclampsia in pregnancy was obtained through linkage to the Medical Birth Registry of Norway . RESULTS: Thirty-three (3%) women developed preeclampsia . The risk of developing preeclampsia seemed to be increased for women who were seronegative for the viruses studied . Seronegativity for Toxoplasma gondii did not show such a pattern . INTERPRETATION: Women who are seronegative for antibodies against viral agents transmitted through close contact seem more likely to develop preeclampsia . This finding indicates that women who are seronegative to such agents may acquire primary infection in pregnancy, and subsequently be at increased risk of preeclampsia . This hypothesis could represent a new approach to the causes of preeclampsia, and encourage search for yet unidentified microbes as a possible causal factor. Nat Immunol, 2001 Dec, 2(12), 1138 - 43 Critical role of IL-15-IL-15R for antigen-presenting cell functions in the innate immune response; Ohteki T et al.; Activation of dendritic cells (DCs) and macrophages by infectious agents leads to secretion of interleukin 12 (IL-12), which subsequently induces interferon-gamma (IFN-gamma) production by multiple cell types that include DCs and macrophages . In turn, IFN-gamma acts on macrophages to augment IL-12 secretion and to produce nitric oxide (NO), which eradicates infected microbes . We show here that in cytokine common gamma subunit-deficient and/or IL-2 receptor beta-deficient mice, production of IL-12, IFN-gamma and NO by DCs and macrophages was severely impaired, as was up-regulation of major histocompatibility complex class II and CD40 . Similar phenotypes were observed in DCs and macrophages from IL-15-deficient mice but not in those from IL-2-deficient mice . This shows that the IL-15-IL-15R interaction is critical in early activation of antigen-presenting cells and plays an important role in the innate immune system. Annu Rev Phytopathol, 2000, 38, 443 - 459 ADVANCES IN IMAGING THE CELL BIOLOGY OF PLANT-MICROBE INTERACTIONS; Heath MC; All plant-microbe interactions are initiated at the level of the cell . Recently, the light microscope has increased in popularity as an investigative tool in plant cell biology, in part because of the parallel developments of confocal laser scanning and video microscopy, computerized image processing, and an ever-increasing array of fluorescent probes that can be applied to living cells . In addition, transgenic plants and cells can be generated in which specific components are fluorescently labeled without any invasive experimental manipulation . The application of such techniques to plant-microbe interactions has revealed microbe-induced changes in cytosolic calcium levels, the visualization of reactive oxygen species generation, cytoskeleton rearrangements, DNA cleavage, and the detailed resolution of intercellular and intracellular trafficking of viral components . These techniques, integrated with electron microscopy, molecular genetics, and other types of investigations, are likely to play an increasingly important role in future studies of plant responses to microbial pathogens or mutualists. J Dent Educ, 2001 Oct, 65(10), 1147 - 53 Clinical decision-making for caries management in root surfaces; Leake JL; This report presents the results of an evidence-based approach to obtaining the best available information on the natural history, prevalence, incidence, diagnosis, and treatment of root caries . Searches of electronic databases produced 807 references; from these and from citations in the selected articles, a final 161 references were used . We found that the information on the natural history of the disease does not provide practitioners with probabilities of, or time estimates for, progression of the disease through stages . For patients aged thirty and older, the prevalence of root caries is roughly 20 to 22 percent less than a person's age . Severity reaches over one lesion by age fifty, two lesions by age seventy, and just over three lesions for those seventy-five and older . About 8 percent (odds of 1:11) of the population would be expected to acquire one or more new root caries lesions in one year . The accuracy of current systems of diagnosis is unknown, although color has been shown to have little validity . Using the criteria of "softness" to define active lesions has been validated by the presence of microbes in the lesion . One strong study and other studies with weaker design or shorter duration add consistent support for the use of fluorides in the remineralization of root caries . Every three-month application of chlorhexidine varnish was shown to be efficacious in one arm of one study . Evidence for restoration of root caries is tentative since the studies were of limited design and duration. Mar Pollut Bull, 2001 Oct, 42(10), 852 - 63 Incorporation of organic tritium (3H) by marine organisms and sediment in the severn estuary/Bristol channel (UK); McCubbin D et al.; Discharges of tritium (3H) into the Severn estuary/Bristol Channel (UK) arise from the authorized release of wastes from nuclear power plants at Hinkley Point and Berkley/Oldbury and from the Nycomed-Amersham radiochemical plant, via the sewer system, at Cardiff . The wastes from the nuclear power plants probably consist almost entirely of 3H2O, whereas those from the radiochemical plant also include uncharacterized 3H labelled organic compounds . The total 3H concentrations in demersal fish and other benthic organisms in the vicinity of the Cardiff Eastern sewer outfall are significantly elevated compared to those observed around other UK nuclear establishments . Concentrations in filtered seawater were approximately 10 Bq kg(-1) whilst levels in surface sediment, seaweed (Fucus vesiculosis) and mussels (Mytilus edulis)/flounder (Platichthys flesus) were in the order of 6 x 10(2), 2 x 10(3), and 10(5) Bq kg(-1) (dry weight), respectively . Almost all the 3H found in sediment and biota were organically bound tritium (OBT) . The high concentration in these materials, relative to that in seawater, is due to the presence of bioavailable organic 3H labelled compounds in the radiochemical waste . It is suggested that bioaccumulation of 3H by benthic organisms and demersal fish occurs primarily via a pathway of physico-chemical sorption/bacterial transformation of dissolved 3H labelled organic compounds into particulate organic matter, and subsequent transfer up a web of sediment dwelling microbes and meiofauna . Variations in 3H accumulation between individual organisms have been interpreted in terms of their different feeding behaviour . Relatively low concentrations were observed in the herbivorous winkle (Littorina littorea) and the pelagic Sprat (Spratus spratus) compared with other benthic organisms and demersal fish . The elevated 3H concentrations in seafood, due to bioaccumulation of OBT, have low radiological significance even for the local critical group of seafood consumers. Ann Chir, 2001 Oct, 126(8), 743 - 50 {Coagulation activation with intraportal islets of Langerhans transplantation in swine}; Lamblin A et al.; STUDY AIM: Intraportal islet allograft appears to be one of the promising treatments for type I diabetes . However, many limiting factors persist . An activation of the coagulation cascade upon contact with islets, has been reported recently in vitro and could play a crucial role in a non specific inflammatory reaction and favour the specific immune reaction . The aim of this experimental study was to confirm in vivo this activation of the coagulation cascade . MATERIAL AND METHODS: An allogenic islets preparation or a material control (inert microbeads) was injected intraportally, in Large White pigs (n = 26), associated with or without an anticoagulant treatment (heparin) . Systemic markers of haemostasis were measured in pigs for 72 hours following injection of the studied material . RESULTS: The thrombin-antithrombin complex increased and platelet count decreased in groups receiving preparation of islets, both indicators of an activation of the coagulation cascade . This activation was proportional to the injected volume and was partially attenuated by heparin . No activation was observed in pigs receiving the material control . CONCLUSION: The activation of the coagulation cascade and the non specific inflammatory reaction could be one of the obstacles to the success of the islet allografts . The use of anticoagulant and anti-inflammatory molecules could potentially allow an improvement of the present results of islet allograft. Andrologia, 2001 Sep, 33(5), 266 - 71 Use of microbeads for the detection of binding sites on the human zona pellucida: a scanning electron microscopy (SEM) assay; Michelmann HW et al.; One prerequisite for fertilization is the specific binding of spermatozoa to the zona pellucida . However, the factors and mechanisms involved in this gamete contact are not well understood . Gamete recognition and binding are species-specific and are controlled by oligosaccharides of the zona and their corresponding carbohydrates on the spermatozoon . By using a specific lectin we developed a technique to detect those oligosaccharides on the human zona pellucida that might be involved in the binding process . Microbeads (O = 2.8 microm), used as artificial spermatozoa, were coated with lectin Con A and cultured together with 75 unfertilized oocytes (group A) remaining after intracytoplasmic sperm injection . Con A binds specifically to alpha-d-mannose and alpha-d-glucose . As a control, 75 unfertilized oocytes after intracytoplasmic sperm injection (group B) were also cultured together with Con A-covered microbeads, but in a medium containing a binding inhibiting sugar (alpha-methyl-mannopyrasosid) . The number and distribution of the microbeads on human oocytes of both groups were analysed on scanning electron microscopy images . Beads on oocytes of group A had binding patterns similar to those of spermatozoa . They were distributed in an extremely heterogeneous way with various numbers of bound beads both on individual and different oocytes . Most of the group A oocytes (85%) had more than 50 beads bound to the zona, in contrast to the control oocytes of group B, where 68% had less than 10 bound beads . The use of an inhibiting sugar abolished the binding capacity of the microbeads nearly completely . This technique is a powerful tool for the detection of binding sites on the zona pellucida, i.e . those sugars that are responsible for contact between spermatozoa and the zona pellucida. Proteomics, 2001 Aug, 1(8), 1033 - 42 Structural modifications of Methanococcus jannaschii flagellin proteins revealed by proteome analysis; Giometti CS et al.; Methanococcus jannaschii is an autotrophic archaeon originally isolated from an oceanic thermal vent . The primary metabolic pathway for energy production in this hyperthermophilic microbe is methanogenesis from H2 and CO2 . As an autotroph, M . jannaschii requires only CO2 as a carbon source for synthesizing all necessary biomolecules . Changes in the environmental availability of these molecules can be expected to activate regulatory mechanisms manifested as the up and down regulation of specific genes and the concomitant increase and decrease in abundance of the corresponding proteins . In our analysis of the proteome of M . jannaschii, we have observed significant changes in the abundance of a common subset of predominant proteins in response to reduced H2 concentration, limited ammonium availability, and the stage of cell growth (exponential compared with stationary) . The masses of tryptic peptides from these proteins match those predicted by M . jannaschii genome open reading frames annotated as flagellin B1 (MJ0891) and flagellin B2 (MJ0892) . Multiple proteins with different isoelectric points and molecular weights match each of these proteins, and the abundance of these protein variants changes with growth conditions . These data indicate that structural modifications altering both the isoelectric point and size of the M . jannaschii flagellin B1 and B2 proteins occur in response to growth conditions and growth stage of M . jannaschii and further suggest the regulation of M . jannaschii motility through structural modifications of the building blocks of the flagella. Int J Med Microbiol, 2001 Sep, 291(4), 251 - 60 Toll-like receptors: cellular signal transducers for exogenous molecular patterns causing immune responses; Kirschning CJ et al.; Innate immunity initiates protection of the host organism against invasion and subsequent multiplication of microbes by specific recognition . Germ line-encoded receptors have been identified for microbial products such as mannan, lipopeptide, peptidoglycan (PGN), lipoteichoic acid (LTA), lipopolysaccharide (LPS), and CpG-DNA . The Drosophila Toll protein has been shown to be involved in innate immune response of the adult fruitfly . Members of the family of Toll-like receptors (TLRs) in vertebrates have been implicated as pattern recognition receptors (PRRs) . Ten TLRs are known and six of these have been demonstrated to mediate cellular activation by distinct microbial products . TLR4 has been implicated as activator of adaptive immunity, and analysis of systemic LPS responses in mice led to the identification of LPS-resistant strains instrumental in its identification as a transmembrane LPS signal transducer . Structural similarities between TLRs and receptor molecules involved in immune responses such as CD14 and the IL-1 receptors (IL-1Rs), as well as functional analysis qualified TLR2 as candidate receptor for LPS and other microbial products . Targeted disruption of the TLR9 gene in mice led to identification of TLR9 as CpG-DNA signal transducer . Involvement of TLR5 in cell activation by bacterial flagellin has been demonstrated . Further understanding of recognition and cellular signaling activated through the ancient host defense system represented by Toll will eventually lead to means for its therapeutic modulation. Med Clin North Am, 2001 Nov, 85(6), 1329 - 47 Host defense in respiratory infections; Welsh DA et al.; Respiratory defenses against infection involve a diverse and complex system . Mechanical barriers limit exposure of the respiratory tract to potential pathogenic organisms, whereas the mucociliary apparatus and cough reflexes work to expel any microbes that may bypass the initial defenses . When microorganisms have gained entry to the lower respiratory tract, the alveolar macrophage and recruited phagocytes may eliminate the culprits before active infection can be established . Only after the failure of the innate immune defenses is a specific immune response mounted . Examination of clinical defects in host defense allows one to understand the importance of the multitude of components of the lung's immune defense system. Appl Environ Microbiol, 2001 Nov, 67(11), 5143 - 53 Bacteria and Archaea physically associated with Gulf of Mexico gas hydrates; Lanoil BD et al.; Although there is significant interest in the potential interactions of microbes with gas hydrate, no direct physical association between them has been demonstrated . We examined several intact samples of naturally occurring gas hydrate from the Gulf of Mexico for evidence of microbes . All samples were collected from anaerobic hemipelagic mud within the gas hydrate stability zone, at water depths in the ca . 540- to 2,000-m range . The delta(13)C of hydrate-bound methane varied from -45.1 per thousand Peedee belemnite (PDB) to -74.7 per thousand PDB, reflecting different gas origins . Stable isotope composition data indicated microbial consumption of methane or propane in some of the samples . Evidence of the presence of microbes was initially determined by 4,6-diamidino 2-phenylindole dihydrochloride (DAPI) total direct counts of hydrate-associated sediments (mean = 1.5 x 10(9) cells g(-1)) and gas hydrate (mean = 1.0 x 10(6) cells ml(-1)) . Small-subunit rRNA phylogenetic characterization was performed to assess the composition of the microbial community in one gas hydrate sample (AT425) that had no detectable associated sediment and showed evidence of microbial methane consumption . Bacteria were moderately diverse within AT425 and were dominated by gene sequences related to several groups of Proteobacteria, as well as Actinobacteria and low-G + C Firmicutes . In contrast, there was low diversity of Archaea, nearly all of which were related to methanogenic Archaea, with the majority specifically related to Methanosaeta spp . The results of this study suggest that there is a direct association between microbes and gas hydrate, a finding that may have significance for hydrocarbon flux into the Gulf of Mexico and for life in extreme environments. Biosens Bioelectron, 2001 Dec, 16(9-12), 1021 - 6 Yeast-immobilized SPV device for koji quality control in sake brewing process; Chiyo T et al.; The malted rice, koji, is an indispensable material for the brewing of sake . It saccharifies rice starch and supplies vitamins for the yeast in sake brewing . Since the quality of sake depends strongly on the quality of koji, quality control of koji is very important in the brewing . There are some methods to measure the activity of enzymes and the quantity of vitamins with the quality of koji . None of these methods, however, directly relate to the yeast metabolism . We constructed a sensor system to monitor the yeast metabolism in sake brewing by use of immobilized Saccharomyces cerevisiae and a Surface PhotoVoltage device (SPV) . In this system, S . cerevisiae K701 and K9, designed for use in sake brewing by the Brewing Society of Japan, were employed as immobilized microbe . The pH change due to the production of organic acids in sake brewing is measured using the SPV . A linear relationship was observed between decrease in the photocurrent (the metabolism response) and the concentration to less than 60 mM of glucose (r=0.990) . Then we measured the koji extract and observed the difference of response between K701 and K9 which corresponded to the productivity of acidic substances by batch test. Immunity, 2001 Oct, 15(4), 579 - 89 Epstein-Barr virus transactivates the human endogenous retrovirus HERV-K18 that encodes a superantigen; Sutkowski N et al.; Superantigens (SAgs) are proteins produced by pathogenic microbes to elicit potent, antigen-independent T cell responses that are believed to enhance the microbes' pathogenicity . Here we show that the human lymphotropic herpesvirus Epstein-Barr virus (EBV) transcriptionally activates the env gene of an endogenous retrovirus, HERV-K18, that possesses SAg activity . SAg activity was demonstrated by MHC class II dependent preferential activation of TCRVB13 T cells in response to murine B cells transfected with the HERV-K18 env gene . This is a unique demonstration of a pathogen inducing a host-encoded Sag and accounts for the previously described EBV associated Sag activity . The T cell activation elicited by the Sag could play a central role in EBV infection and associated diseases. Respir Res, 2000, 1(3), 129 - 32 Epub 2000 Oct 25. Mimicking microbial 'education' of the immune system: a strategy to revert the epidemic trend of atopy and allergic asthma? Matricardi PM, Bonini S. Deficient microbial stimulation of the immune system, caused by hygiene, may underly the atopy and allergic asthma epidemic we are currently experiencing . Consistent with this 'hygiene hypothesis', research on immunotherapy of allergic diseases also centres on bacteria-derived molecules (eg DNA immunostimulatory sequences) as adjuvants for allergen-specific type 1 immune responses . If we understood how certain microbes physiologically 'educate' our immune system to interact safely with environmental nonmicrobial antigens, we might be able to learn to mimic their beneficial actions . Programmed 'immunoeducation' would consist of safe administration, by the correct route, dose and timing, of those microbial stimuli that are necessary to 'train' the developing mucosal immune system and to maintain an appropriate homeostatic equilibrium between its components . Overall, this would result in a prevention of atopy that is not limited to certain specific allergens . Although such a strategy is far beyond our present potential, it may in principle revert the epidemic trend of atopy and allergic asthma without jeopardizing the fight against infectious diseases. J Am Dent Assoc, 2001 Sep, 132(9), 1241 - 5 The effects of toothpastes on the residual microbial contamination of toothbrushes; Warren DP et al.; BACKGROUND: Contaminated toothbrushes have been shown to harbor and transmit viruses and bacteria . The authors conducted a study to evaluate the effect of a triclosan-containing toothpaste on the residual anaerobic microbial contamination of toothbrushes . METHODS: Twenty patients who had Type III or Type IV periodontitis participated in this study . One side of each of their mouths served as a control (no toothpaste) . The teeth on the other side were brushed with a regular toothpaste or a triclosan-containing toothpaste . After the toothbrushes were allowed to dry in air for four hours, the authors placed the toothbrush heads in solution, dislodged the microbes from the brushes by vortexing and plated them in culture dishes . The authors anerobically incubated the culture dishes and determined the presence or absence of Prevotella species or Ps; Porphyromonas gingivalis, or Pg; and Actinobacillus actinomycetemcomitans, or Aa . RESULTS: The authors detected Aa and Pg on the control toothbrushes more frequently than they did Ps . This variation in isolation frequency was statistically significant by chi 2 analysis (P < .001) . The authors compared the isolation frequency of the three test organisms between the control and regular-toothpaste groups, between the control and triclosan-containing--toothpaste groups, and between the triclosan-containing--toothpaste and regular-toothpaste groups . They found no significant intergroup differences in the isolation frequencies after using chi 2 analysis . CONCLUSIONS: Toothpaste use reduced the residual microbial contamination for two of three test organisms, but the lower isolation frequencies were not statistically significant . Further study in this area is indicated . CLINICAL IMPLICATIONS: Dental professionals should advise patients who have systemic, localized or oral inflammatory diseases to disinfect or frequently replace their toothbrushes. Science . 1985 Jan 11;227(4683):153. Monsanto may bypass NIH in microbe test; Sun M; KIE: The Monsanto Company is planning to ask the Environmental Protection Agency for clearance to field test a genetically engineered microbial pesticide, bypassing the traditional approval process of the National Institutes of Health . Although only federally funded institutions are required to obtain NIH approval for genetic engineering tests, Monsanto is the first company to bypass the NIH regulatory process, which has become mired in a lawsuit brought by Jeremy Rifkin . Ukr Biokhim Zh, 2001 Mar-Apr, 73(2), 130 - 7 {Features of glutamate dehydrogenase in fetal and adult rumen tissue}; Kalachniuk HI et al.; Glutamate dehydrogenase (GDH) from rumen mucosa of cow fetus, liver and two forms from mucosa (bacterial and tissue) of the adult animal were partly purified and characterized . The activity of the bacterial glutamate dehydrogenase was shown to depend on qualities of a biomass of microbes, adhered on surface of rumen mucosa . All enzymes from tissues (GDHTRF, TRC, TLC), revealed the hypersensibility to increase in the concentration medium of Zn2+, guanosine triphosphate (GTP), acting here in a role of negative modulators, and also adenosine monophosphate (AMP) and leucine, which acted as activators . However, in the same concentrations these effectors do not influence the activity of the bacterial glutamate dehydrogenase . And if all tissues enzymes are highly specific to coenzyme NADH, the bacterial ones almost in 3 times is more active at NADPH use. Int Arch Allergy Immunol, 2001 Sep, 126(1), 23 - 31 Some aspects of complementarity in the immune system . A bird's eye view; Bakacs T et al.; The burden of this paper is the suggestion that the defence capacity of the immune system is rather limited . It cannot stand in readiness to deal with a practically endless diversity and abundance of microbes . In contrast to conventional thinking the current model proposes: (1) The core idea that cells of the immune system are basically and constantly interconnected with host cells (e.g., through TCR-MHC interactions) and that foreign antigens (peptides) may tend to obstruct such interactions . Peptides presented during a viral infection typically decrease complementarity between the structures that are the products of the major histocompatibility complex (MHC) genes (or other genes related to it) and T cells . The altered MHC profile exposes infected cells to a polyclonal immune attack from other T cells such that tissue destruction occurs in an allograft rejection-like fashion . This may explain why a substantial portion of T cell numbers is activated when only a small number of specific T cells is 'obstructed' from functioning by the presence of nonself peptides . (2) Phagocytes 'see' targets even in a non-immune host because complement distribution associated with polyreactive natural antibodies magnifies sensitization differences between pathogens and host cells . (3) There is only a probability that hypermutation will successfully change the genome in some B cell clones to produce high affinity antibodies that prevent the re-infection of the host by the same pathogen, but cannot conquer primary infections . (4) The history of the development of the immune responses suggests that during prolonged interaction between host and microbes in our natural habitat, carried on over many generations, the adaptive antibody population may facilitate the evolution of the natural antibody repertoire . The model predicts that microbes, which are not a part of the local environment, may invade the organism without significant resistance . The model is discussed in various interactions for survival in the context of infection and tumorigenicity . Dan Medicinhist Arbog . 1998;:61-76. Not Available Rasmussen L. Louis Pasteur in France saw that microbes might cause disease, Gerhard Armauer Hansen in Norway established a connection between a bacterium and a human disease, and Robert Koch in Germany supplied the final proof that linked bacteria to disease . From then on there was an explosion in interest and techniques for cultivation of bacteria . It lasted about 30 years before Harrison in the USA succeeded in growing cells from multicellular organisms in pure culture . His technique came to Denmark at an early time point and the Rockefeller Foundation in New York and the Carlsberg Foundation in Copenhagen joined forces and built the Biological Institute of the Carlsberg Foundation in order to support Albert Fischer in his study of cell biology . His first assistant at this Institute, Fritz Lipmann, won the Nobel Prize for 1953 for work initiated there . The Institute remained a center for cell biology for almost 50 years . At the neighboring Rockefeller Institute in Copenhagen Einar Lundsgaard obtained results showing in the short perspective that current concepts of cellular utilization of energy were wrong, and in the long perspective his assistant, Herman Kalckar, would formulate modern concepts of cellular oxidative phosphorylations . This review is based in part on films from the 1930's. Ber Wissenschaftsgesch, 1996, 19(2-3), 81 - 94 Not Available Gradmann C. The text analyses metaphors of bacteriology which were extensively used in Germany during the era of William II . These display--in a vivid exchange with the scientific concepts of the age--a specific popular understanding of disease based on bacteriology . Disease is essentially seen as a war of physicians against microbes . While popularizing science bacteriological metaphors became part of the political language of their age . At the same time the prestige of bacteriology was in turn employed to lend credibility to pictures of assumed enemies--by portraying them as infectious diseases . Although the political language of bacteriology differed from the social darwinism of the age in important structural and semantic aspects, it nevertheless influenced the political language of its time, for example by becoming a blueprint of antisemitic rhetorics. Hist Cienc Saude Manguinhos, 1995, 2(1), 67 - 98 {Domingos José Freire and the beginnings of bacteriology in Brazil}; Benchimol JL; The article examines the trajectory of Domingos Jose Freire, a bacteriologist who attained great national and international renown in the final quarter of the last century for claiming to have discovered the yellow-fever microbe and a vaccine that was administered to thousands of people . The article reviews the main controversies protagonized by this scientist and endeavors to show the role be played in the enthronement of Pasteurian medicine in Brazil. Zhonghua Yi Shi Za Zhi, 1999 Oct, 29(4), 203 - 6 {A brief history of discovery of Helicobacter pylori}; Li J et al.; The theoretical background for the diagnosis and treatment of upper digestive ulcer, gastritis and stomach carcinema was challenged by H . pylori, a microbe seems only under the microscope . This paper makes a brief review on the brief history of discovery and recent conclusion on the microbe, and explores its challenge to the traditional conception. Yeni Tip Tarihi Arastirmalari, 1998, 4, 237 - 43 {Some medical documents written by Ahmed Cevdet Pasha}; Izgoer AZ; The assumption that civilization was originated in the East, was one of the most important points emphasized by the intelligentsia of the late Ottoman reign . This conviction was appreciated in the thought of Ahmed Cevdet Pasha (1823 - 1895), a famous figure of the period . Cevdet Pasha, who believed that the source of science and education was started in the East, claimed that Europe was acquainted with Islamic science during the Crusades . In this paper Cevdet Pasha's ideas related with small-pox, cholera, plague, poisons and microbes are mentioned . This study is based on Cevdet Pasha's documents related with these subjects; and the transcriptions of the texts are given. Proc Natl Acad Sci U S A, 1996 May 14, 93(10), 4600 - 3 Mapping leaf surface landscapes; Mechaber WL et al.; Leaf surfaces provide the ecologically relevant landscapes to those organisms that encounter or colonize the leaf surface . Leaf surface topography directly affects microhabitat availability for colonizing microbes, microhabitat quality and acceptability for insects, and the efficacy of agricultural spray applications . Prior detailed mechanistic studies that examined particular fungi-plant and pollinator-plant interactions have demonstrated the importance of plant surface topography or roughness in determining the outcome of the interactions . Until now, however, it has not been possible to measure accurately the topography--i.e., the three-dimensional structure--of such leaf surfaces or to record precise changes in patterns of leaf surface elevation over time . Using contact mode atomic force microscopy, we measured three-dimensional coordinates of upper leaf surfaces of Vaccinium macrocarpon (cranberry), a perennial plant, on leaves of two age classes . We then produced topographic maps of these leaf surfaces, which revealed striking differences between age classes of leaves: old leaves have much rougher surfaces than those of young leaves . Atomic force microscope measurements were analyzed by lag (1) autocorrelation estimates of leaf surfaces by age class . We suggest that the changes in topography result from removal of epicuticular lipids and that the changes in leaf surface topography influence phylloplane ecology . Visualizing and mapping leaf surfaces permit detailed investigations into leaf surface-mediated phenomena, improving our understanding of phylloplane interactions. Proc Natl Acad Sci U S A, 1990 Oct, 87(19), 7565 - 9 Sterol phylogenesis and algal evolution; Nes WD et al.; The stereochemistry of several sterol precursors and end products synthesized by two fungal-like micro-organisms Prototheca wickerhamii (I) and Dictyostelium discoideum (II) have been determined by chromatographic (TLC, GLC, and HPLC) and spectral (UV, MS, and 1H NMR) methods . From I and II the following sterols were isolated from the cells: cycloartenol, cyclolaudenol, 24(28)-methylenecycloartanol, ergosterol, protothecasterol, 4alpha-methylergostanol, 4alpha-methylclionastanol, clionastanol, 24beta-ethylcholesta-8,22-enol, and dictyosterol . In addition, the mechanism of C-24 methylation was investigated in both organisms by feeding to I {2-3H}lanosterol, {2-3H}cycloartenol, {24-3H}lanosterol, and {methyl-2H3}methionine and by feeding to II {methyl-2H3}methionine . The results demonstrate that the 24beta configuration is formed by different alkylation routes in I and II . The Delta25(27) route operates in I while the Delta24(28) route operates in II . Based on what is known in the literature regarding sterol distribution and phylogenesis together with our findings that the stereochemical outcome of squalene oxide cyclization leads to the production of cycloartenol rather than lanosterol (characteristic of the fungal genealogy) and the chirality of the C-24 alkyl group is similar in the two nonphotosynthetic microbes (beta oriented), we conclude that Prototheca is an apoplastic Chlorella (i.e., an alga) and that Dictyostelium as well as the other soil amoebae that synthesize cycloartenol evolved from algal rather than fungal ancestors. Prikl Biokhim Mikrobiol, 2001 Sep-Oct, 37(5), 607 - 11 {Effect of proteinaceous polygalacturonase inhibitors from apple seed tissue on an enzyme isolated from phytopathogenic fungi}; Glinka EM et al.; A protein polygalacturonidase inhibitor isolated from fruit of the apple varieties Antonovka and Mantuanskoe differently affects the polygalacturonidases of different phytopathogenic fungi . Three groups of fungi were recognized by the sensitivity of their polygalacturonidases to the inhibitory effect . Storage of apples after harvesting is accompanied by changes in the inhibitor activity, and the time pattern of these changes depends on the variety . An increase in the inhibitor activity occurs concurrently with the elevation in ethylene release characteristic of the stage of elevated respiration (a climacteric increase) . The data suggest that a decrease in the apple fruit resistance to microbes at the end of the storage period is related, along with other reasons, to a change in the activity of the protein polygalacturonidase inhibitor. Radiat Res, 2001 Nov, 156(5 Pt 2), 612 - 7 The potential impact of bystander effects on radiation risks in a Mars mission; Brenner DJ et al.; Densely ionizing (high-LET) galactic cosmic rays (GCR) contribute a significant component of the radiation risk in free space . Over a period of a few months-sufficient for the early stages of radiation carcinogenesis to occur-a significant proportion of cell nuclei will not be traversed . There is convincing evidence, at least in vitro, that irradiated cells can send out signals that can result in damage to nearby unirradiated cells . This observation can hold even when the unirradiated cells have been exposed to low doses of low-LET radiation . We discuss here a quantitative model based on the a formalism, an approach that incorporates radiobiological damage both from a bystander response to signals emitted by irradiated cells, and also from direct traversal of high-LET radiations through cell nuclei . The model produces results that are consistent with those of a series of studies of the bystander phenomenon using a high-LET microbeam, with the end point of in vitro oncogenic transformation . According to this picture, for exposure to high-LET particles such as galactic cosmic rays other than protons, the bystander effect is significant primarily at low fluences, i.e., exposures where there are significant numbers of untraversed cells . If the mechanisms postulated here were applicable in vivo, using a linear extrapolation of risks derived from studies using intermediate doses of high-LET radiation (where the contribution of the bystander effect may be negligible) to estimate risks at very low doses (where the bystander effect may be dominant) could underestimate the true risk from low doses of high-LET radiation . It would be highly premature simply to abandon current risk projections for high-LET, low-dose radiation; however, these considerations would suggest caution in applying results derived from experiments using high-LET radiation at fluences above approximately 1 particle per nucleus to risk estimation for a Mars mission. Radiat Res, 2001 Nov, 156(5 Pt 1), 526 - 34 Low-dose hypersensitivity in Chinese hamster V79 cells targeted with counted protons using a charged-particle microbeam; Schettino G et al.; The Gray Laboratory charged-particle microbeam has been used to assess the clonogenic ability of Chinese hamster V79 cells after irradiation of their nuclei with a precisely defined number of protons with energies of 1.0 and 3.2 MeV . The microbeam uses a 1-microm silica capillary collimator to deliver protons to subcellular targets with high accuracy . The detection system is based on a miniature photomultiplier tube positioned above the cell dish, which detects the photons generated by the passage of the charged particles through an 18-microm-thick scintillator placed below the cells . With this system, a detection efficiency of greater than 99% is achieved . The cells are plated on specially designed dishes (3-microm-thick Mylar base), and the nuclei are identified by fluorescence microscopy . After an incubation period of 3 days, the cells are revisited individually to assess the formation of colonies from the surviving cells . For each energy investigated, the survival curve obtained for the microbeam shows a significant deviation below 1 Gy from a response extrapolated using the LQ model for the survival data above 1 Gy . The data are well fitted by a model that supports the hypothesis that radioresistance is induced by low-dose hypersensitivity . These studies demonstrate the potential of the microbeam for performing studies of the effects of single charged particles on cells in vitro . The hypersensitive responses observed are comparable with those reported by others using different radiations and techniques. J Am Chem Soc, 2001 Oct 24, 123(42), 10173 - 82 Hydroaromatic equilibration during biosynthesis of shikimic acid; Knop DR et al.; The expense and limited availability of shikimic acid isolated from plants has impeded utilization of this hydroaromatic as a synthetic starting material . Although recombinant Escherichia coli catalysts have been constructed that synthesize shikimic acid from glucose, the yield, titer, and purity of shikimic acid are reduced by the sizable concentrations of quinic acid and 3-dehydroshikimic acid that are formed as byproducts . The 28.0 g/L of shikimic acid synthesized in 14% yield by E . coli SP1.1/pKD12.138 in 48 h as a 1.6:1.0:0.65 (mol/mol/mol) shikimate/quinate/dehydroshikimate mixture is typical of synthesized product mixtures . Quinic acid formation results from the reduction of 3-dehydroquinic acid catalyzed by aroE-encoded shikimate dehydrogenase . Is quinic acid derived from reduction of 3-dehydroquinic acid prior to synthesis of shikimic acid? Alternatively, does quinic acid result from a microbe-catalyzed equilibration involving transport of initially synthesized shikimic acid back into the cytoplasm and operation of the common pathway of aromatic amino acid biosynthesis in the reverse of its normal biosynthetic direction? E . coli SP1.1/pSC5.214A, a construct incapable of de novo synthesis of shikimic acid, catalyzed the conversion of shikimic acid added to its culture medium into a 1.1:1.0:0.70 molar ratio of shikimate/quinate/dehydroshikimate within 36 h . Further mechanistic insights were afforded by elaborating the relationship between transport of shikimic acid and formation of quinic acid . These experiments indicate that formation of quinic acid during biosynthesis of shikimic acid results from a microbe-catalyzed equilibration of initially synthesized shikimic acid . By apparently repressing shikimate transport, the aforementioned E . coli SP1.1/pKD12.138 synthesized 52 g/L of shikimic acid in 18% yield from glucose as a 14:1.0:3.0 shikimate/quinate/dehydroshikimate mixture. Dev Comp Immunol, 2001 Oct-Dec, 25(8-9), 725 - 43 The acute phase response and innate immunity of fish; Bayne CJ et al.; Tissue trauma or invasion by pathogens or parasites induce changes in the quantities of several macromolecules in animal body fluids . These changes comprise one aspect of the acute phase response (APR), which in toto involves metabolic changes in several organ systems . One clear indication of the response is the increase in synthesis and secretion by the liver of several plasma proteins, with simultaneous decreases in others . These acute phase proteins (APP) function in a variety of defense-related activities such as limiting the dispersal of infectious agents, repair of tissue damage, inactivation of proteases, killing of microbes and other potential pathogens, and restoration of the healthy state . Some APP are directly harmful to microbes, while others modify targets thus marking them for cell responses . Some work alone while others contribute to cascades . Proteins that are APP in mammals, and that have been identified in both teleosts and elasmobranchs include C-reactive protein, serum amyloid P, and several components of the Complement system . Others reported in teleosts include transferrin and thrombin . Of these, only CRP has been reported to increase in acute phase plasma . In trout, a precerebellin-like protein is an APP with unknown functions . A cDNA library enriched in fragments of transcripts that were more abundant in livers from fish undergoing an APR recently yielded sequences resembling 12 additional known APP, and as many others either not known to be APP, or not similar to others yet in public databases . It appears that, as in mammals, hepatocytes are the prime source of APP in fish, and that pro-inflammatory cytokines induce transcription of their genes. Med Hypotheses, 2001 Oct, 57(4), 484 - 6 Control of skin infections by a combined action of ultraviolet A (from sun or UVA lamp) and hydrogen peroxide (HUVA therapy), with special emphasis on leprosy; Ahmad SI; Despite its abundance and certain therapeutic value, the importance of sunlight in the treatment of infectious skin diseases has not been fully exploited . One reason is that a sufficient amount of the damaging components of sunlight (UVC and most UVB) cannot reach us and the band of UV that can reach (UVA) is a poor inactivator of living cells . UVA, however, can be deleterious to cells in the presence of sensitizers and a number of biological and chemical sensitizers have been identified which can inactivate microbes in the presence of UVA.Of several known agents, I have selected hydrogen peroxide (H(2)O(2)) as a UVA sensitizer and propose that a combined action of H(2)O(2)and UVA (HUVA therapy) can be utilized in controlling skin infections of various types.Of particular interest is infection by Mycobacterium leprae, which is known to affect many millions of humans globally . H(2)O(2)being relatively cheap (and UVA from the sun being free) the cost of application, particularly in third-world countries where leprosy is more common, would be low and therefore the treatment can be employed on a wide scale . A further reason for proposing the use of H(2)O(2)is that, out of several agents we have tested, this was found to be the most potent; it is also easily able to reach target sites, very cheap, relatively safe and there is no known microbial resistance to HUVA . Rom J Virol, 1999 Jan-Dec, 50(1-4), 53 - 70 Evidence of TNF system activation and high expression of TNF alpha messenger transcripts in necrotizing enterocolitis of HIV-infected children; Ispas D; TNF alpha contributes to the necrotizing enterocolitis (NEC) pathogenesis . To date, this clinical entity of neonates was never described in HIV-infected children . In 15 HIV-positive children with histological evidence of various intestinal lesions resembling NEC, we have studied serum TNF alpha and soluble TNF receptor concentrations by ELISAs, and archived paraffin embedded intestinal tissues by in situ hybridization with DIG-labeled RNA probes for TNF alpha messenger transcripts . We found increased levels of TNF alpha and soluble receptors, proving TNF alpha system activation . We detected TNF alpha messenger transcripts in all cases, regardless of the presence of microbial pathogens at intestinal level . Since HIV can infect many cells of the gastrointestinal tract, also triggering the secretion of TNF alpha, we concluded that factors contributing to NEC pathogenesis in HIV-infected children are complex . At least the nutritional and immunological status are involved, other viral co-infections, opportunistic microbes (such as mycobacteria), and pathogenic activities of HIV . All together enhance both circulating TNF alpha system and its cytotoxic effects at intestinal level. Burns, 2001 Nov, 27(7), 699 - 707 Strategies to improve the take of commercially available collagen/glycosaminoglycan wound repair material investigated in an animal model; Grant I et al.; Integra Artificial Skin is becoming widely used in burns and reconstructive surgery . However, poor take and loss due to infection remains a concern for some patients . This study describes how the successful bio-integration of Integra Artificial Skin was accomplished in isolated full-thickness wounds in the pig, by the use of sequential protocol modifications that more intimately opposed the material to the wound bed . Further improvement was achieved by measures to reduce bacterial colonisation including the use of topical anti-microbial agents . The level of histologically confirmed Integra 'take' improved from close to zero to a mean 96% of wound area (+/-11 S.D.) . The efficacy of this dermal wound bed was confirmed by the histological analysis of wounds after the subsequent application of extremely thin (<130 microm) widely meshed autologous split-thickness skin grafts . The results of this animal study support the recommendation that the bio-integration of Integra Artificial Skin is best achieved by the elimination of dead space below the material and by measures to hinder the penetration of microbes . The significant improvement in take achieved in this animal model using simple methodological refinements re-identifies priorities for the use of Integra Artificial Skin that can be applied in clinical management. Environ Toxicol Chem, 2001 Oct, 20(10), 2287 - 95 Pollution-induced community tolerance and functional redundancy in a decomposer food web in metal-stressed soil; Salminen J et al.; Pollution may lead to the development of pollution-induced community tolerance (PICT) in a stressed community . We studied the presence of PICT in soil food webs using soil microcosms . Soil microcosms containing soil invertebrates and microbes were collected from polluted and unpolluted areas and exposed to a range of soil zinc concentrations . A pine seedling was planted in each microcosm to measure the effects of the origin of the community and Zn pollution on above-ground plant production . The effects of the treatments on nutrient content in the soil were also measured . The diversity of soil microarthropods and the soil's mineral nutrient content were low at the Zn-polluted site . We did not observe an increasing Zn tolerance among the soil organisms in the polluted soil . However, low population growth rates of soil invertebrates from the polluted site may indicate the deleterious effects on fitness of long-lasting pollution . In the soil from the nonpolluted site, Zn additions caused changes in the invertebrate food web structure . These changes were explained by the good physiological condition of the animals and their insensitivity to Zn . The fact that the food web structure in soil from the polluted site did not change can be used as a rough indicator of PICT . Structural stability is presumed by the lack of Zn-sensitive species at this site and the inability of populations to acclimate by altering their growth or reproduction patterns in response to changing soil conditions . Although microbial-based soil decomposer systems may have a high functional redundancy, our results indicate that metal stress at the polluted site exceeds the tolerance limits of the system . As a consequence, ecosystem function at this site is endangered . This study also shows that the evolution of metal tolerance by soil decomposer organisms may not be a common reaction to soil pollution, although changes of population and community structure indicated severe metal stress on organisms. Acta Astronaut, 2001 Jul, 49(2), 105 - 8 Effect of volatile metabolites of dill, radish and garlic on growth of bacteria; Tirranen LS et al.; In a model experiment plants were grown in sealed chambers on expanded clay aggregate under the luminance of 150 W/m2 PAR and the temperature of 24 degrees C . Seven bacterial strains under investigation, replicated on nutrient medium surface in Petri dishes, were grown in the atmosphere of cultivated plants . Microbial response was evaluated by the difference between colony size in experiment and in control . In control, bacteria grew in the atmosphere of clean air . To study the effects of volatile metabolites of various plant on microbial growth, the experimental data were compared with the background values defined for each individual experiment . Expanded clay aggregate, luminance, temperature, and sealed chamber (without plants) for the background were the same . Volatile metabolites from 28-days old radish plants have been reliably established to have no effect on the growth of microbes under investigation . Metabolites of 30-days old dill and 50-days old garlic have been established to have reliable bacteriostatic effect on the growth of three bacterial strains . Dill and garlic have been found to have different range of effects of volatile substances on bacterial growth . Volatile metabolites of dill and garlic differed in their effect on the sensitivity spectrum of bacteria . An attempt has been made to describe the obtained data mathematically . c 2001 International Astronautical Federation . Published by Elsevier Science Ltd. Analyst, 2001 Sep, 126(9), 1545 - 50 Detection of aflatoxins (G(1-2), B(1-2)), sterigmatocystin, citrinine and ochratoxin A in samples contaminated by microbes; Tuomi T et al.; A method is described for the simultaneous determination of common aflatoxins (G1, G2, B1, B2) and their precursor sterigmatocystin, and also citrinine and ochratoxin A . The method was applied to a building material matrix artificially contaminated with mycotoxin-producing fungi . The method includes extraction, sample pre-treatment and reversed-phase HPLC separation with tandem mass spectrometric identification and quantification using electrospray ionisation on a quadrupole ion trap mass analyser (ESI-MS-MS) . Aqueous methanol was used in the initial extraction and solvent partitioning and solid phase extraction in the purification of samples . The HPLC separation was run on-line with the ESI-MS-MS detection . The limit of quantification of the procedure was 200 ng for all compounds . Recoveries of the sample pre-treatment varied from 28 to 99% . The average compound- and concentration-dependent accuracy and precision (RSD) were 21 and 113%, respectively . The method includes small sample volumes (approximately 1 g in 20 ml) and few, non-labour intensive, sample treatment steps . It should allow for a high throughput of samples with good prospects of automation. J Biol Chem, 2001 Dec 14, 276(50), 47143 - 9 Epub 2001 Oct 04. Molecular cloning and functional characterization of chicken toll-like receptors . A single chicken toll covers multiple molecular patterns; Fukui A et al.; Toll-like receptors (TLR) in the innate immune system have not been identified in non-mammalian vertebrates . Two types of TLR were cloned from a chicken bursa cDNA library using degenerate primers based on the consensus sequences of mouse and Drosophila Toll and designated as chicken TLR (chTLR) type 1 and type 2 . Of the nine human TLRs reported to date, these chTLRs showed the highest homology to human TLR2 . The extracellular regions of type 1 and type 2 contained a distinct approximately 200-amino acid stretch and were 45.3 and 46.3% homologous to that of human TLR2 . The intracellular Toll/interleukin-1R homology domain of type 1 and type 2 was perfectly identical to each other and highly homologous (80.7%) to that of human TLR2 . Both types were widely detected by reverse transcriptase-polymerase chain reaction and immunoblotting in various chicken organs, especially those rich in connective tissue . Both genes were mapped to chromosome 4q1.1, suggesting that they arose by gene duplication . By reporter gene assay, type 2 and to a lesser extent type 1, selectively signaled the presence of mycoplasma macrophage-activating lipopeptide-2/M161Ag in the human embryonic kidney 293 cell system . Cotransfection of type 2 and human CD14 or MD-2 into human embryonic kidney 293 cells allowed the response to Escherichia coli lipopolysaccharide (LPS), whereas type 1 did not signal LPS or any other microbial components tested . These results indicated that chTLR type 2 covers two major microbe patterns, lipoproteins and LPS, which are regulated by TLR2 and TLR4 in mammals . In oviparous animals, the duplicated TLRs in the pattern-recognition system may function for host-pathogen discrimination in a manner that is distinct from that in mammals. Eur Respir J, 2001 Sep, 18(3), 571 - 88 Lung mucosal immunity: immunoglobulin-A revisited; Pilette C et al.; Mucosal defence mechanisms are critical in preventing colonization of the respiratory tract by pathogens and penetration of antigens through the epithelial barrier . Recent research has now illustrated the active contribution of the respiratory epithelium to the exclusion of microbes and particles, but also to the control of the inflammatory and immune responses in the airways and in the alveoli . Epithelial cells also mediate the active transport of polymeric immunoglobulin-A from the lamina propria to the airway lumen through the polymeric immunoglobulin receptor . The role of IgA in the defence of mucosal surfaces has now expanded from a limited role of scavenger of exogenous material to a broader protective function with potential applications in immunotherapy . In addition, the recent identification of receptors for IgA on the surface of blood leukocytes and alveolar macrophages provides an additional mechanism of interaction between the cellular and humoral immune systems at the level of the respiratory tract. Eur Respir J, 2001 Sep, 18(3), 542 - 8 Inflammatory mediators in nasal lavage, induced sputum and serum of employees with rheumatic and respiratory disorders; Roponen M et al.; Exposure to microbes present in mould-damaged buildings has been linked to increased frequency of various inflammatory diseases . The current study examined differences in inflammatory mediators in nasal lavage (NAL), induced sputum (IS) and serum of occupants with rheumatic or respiratory disorders and their controls, all working in the same moisture-damaged building . Exhaled nitric oxide (NO) measurements, lung function tests, skin-prick tests and health data collection by questionnaire were performed . Concentrations of NO, interleukin (IL)-1, IL-4, IL-6 and tumour necrosis factor-alpha in NAL, IS and serum (excluding NO and IL-1) of the subjects were measured during an occupational exposure period and the vacation period without such exposure . The concentrations of IL-4 in NAL fluid were significantly higher among all occupants during the working period (geometric mean 8.5 microg x mL(-1), range 0-206.5 microg x mL(-1)), as compared to that during vacation (0.4 microng x mL(-1) range 0-3.7 pg x mL(-1)) (p = 0.008) . Absence from the work environment also significantly diminished reporting of symptoms . IL-4 levels in the serum of case subjects were significantly higher than in controls . Moreover, employees with respiratory symptoms had markedly higher exhaled NO values than their controls (p = 0.028) . In summary, these data suggest that mediators in nasal lavage samples reflect the occupational exposure to moulds, whereas possible indicators of existing disorders are detectable in serum. Biol Sci Space, 2000 Dec, 14(4), 363 - 71 {Biomineralization at hot springs and mineral springs, and their significance in relation to the Earth's history}; Akai J; Recently, there is strong interest on microbe-mineral interactions . This is related also to recent expanded knowledges on extremely severe environments in which microbes live . Interaction between microbes and minerals contains biomineralization processes . Varieties of biomineralization products are found not only in various geologic materials and processes in the earth's history but also in present surface environments . Some hot springs represent such environments similar to those of unique and extremely severe environments for life . In this short review, the author briefly shows some examples of biomineralizations at some hot springs and mineral springs, Japan . In such environments, iron ore was formed and some varieties of growing stromatolites were found . The varieties of stromatolite are siliceous, calcic and manganese types . Cyanobacteria and the other bacteria are related to form the stromatolite structure . In the Gunma iron ore, sedimentary iron ores were mineralogically described in order to evaluate the role of microorganisms and plants in ore formation . The iron ore is composed of nanocrystalline goethite . Algal fossils are clearly preserved in some ores . Various products of biomineralization are found in the present pH 2-3, Fe2(+)- and SO4(2-)-rich streams . Bacterial precipitation had variations from amorphous Fe-P-(S) precipitates near the outlet of mineral spring, to Fe-P-S precipitates and to Fe-S-(P) precipitates . Mosses and green algae are also collecting Fe precipitates in and around the living and dead cells . The Gunma Iron Ore can be said as Biologically Induced Iron Ore . At Onikobe and Akakura hot springs, growing stromatolites of siliceous and calcareous types, were found, respectively . At Onikobe, The stromatolites grow especially near the geyser . Cyanobacterial filaments in stromatolite were well preserved in the siliceous and calcic stromatolites . The filaments oriented in two directions which form the layered structures were found . At Yunokoya hot spring, black and brittle stromatolitic structures which were composed of amorphous Mn minerals are growing . The form of these structures are hemispherical . Many bacteria that were coated with amorphous Mn minerals were found on these structures . Furthermore, Precambrian (Proterozoic : Wittenoom-Chichester region, western Australia) manganese stromatolite was briefly shown in comparison . The black stromatolite has been clarified to be composed of todorokite . Small spotty and donuts-like shaped todorokite aggregates which are very similar to biologically induced Mn-precipitates were found in massive dolomite layers. J Allergy Clin Immunol, 2001 Oct, 108(4 Suppl), S117 - 20 Overview of infections and cardiovascular diseases; Vercellotti GM; Microbes have been proposed as inciting agents of tissue injury and inflammation, both of which underlie the pathogenesis of atherosclerosis . Viruses, including the herpes simplex virus and cytomegalovirus, as well as bacteria such as Chlamydia pneumoniae, have been implicated in the process . In vitro, these agents promote a proinflammatory and a procoagulant phenotype in vascular cells . Viruses augment cell accumulation through alterations of apoptosis . Infectious agents may play a role in pathogenesis of atherosclerosis by triggering an autoimmune response due to microbial molecular mimicry . It is unlikely that a single agent is the sole cause or modulator of this heterogeneous disease . Contradictory epidemiological studies may be reconciled with a new construct suggesting that multiple pathogens infecting an individual in aggregate may promote an inflammatory and procoagulant environment that underlies the pathogenesis of atherosclerosis. J Comp Physiol {B}, 2001 Aug, 171(6), 441 - 7 Influences of incubation temperature and various saccharides on the production of organic acids and gases by gut microbes of rainbow trout Oncorhynchus mykiss in a micro-scale batch culture; Kihara M et al.; We studied the influence of incubation temperature and additional saccharides on the metabolism of hindgut microbes of the rainbow trout Oncorhynchus mykiss in a 50 microl-scale batch culture system . Intestinal contents of rainbow trout reared at 15 degrees C were incubated with glucose, lactosucrose, sodium alginate or colloidal chitin (each 10 g/l) at 15 degrees C or 25 degrees C for 12 h . Levels of organic acids at 0 h and 12 h of incubation were quantified with HPLC . We also monitored gas release from these cultures during incubation . The main product was iso-butyric acid, except for the cultures with colloidal chitin where no net production of organic acids was observed . We detected higher levels of iso-butyric acid in cultures with lactosucrose than in the other cultures . Net production of this acid was less in cultures with colloidal chitin than in blank cultures . The volume of released gas was larger when incubated at 25 degrees C than at 15 degrees C . Cultures with colloidal chitin released more gas than blank cultures when they were incubated at 15 degrees C . Cultures with sodium alginate released less gas than blank cultures irrespective of incubation temperature . These results indicate that the hindgut microbes of this carnivorous fish mainly produce branched-chain fatty acids, very likely by microbial digestion of nitrogenous materials rather than saccharides . However, additional saccharides affected production of branched-chain fatty acids . The influence of incubation temperature in the present study also suggested that the environmental temperature of host fish should affect microbial digestion in the fish gut. Ann Occup Hyg, 2001 Oct, 45(7), 577 - 83 Assessment of exposure to organic dust in a hemp processing plant; Fishwick D et al.; The aim of this preliminary study was to assess exposure to various constituents of the organic dust generated during the processing of hemp in a small group of exposed workers . Airborne levels of inhalable dust, endotoxin and soluble protein, and the respirable, thoracic and inhalable fractions of fungal, bacterial and actinomycete contamination were measured in the personal breathing zone of exposed workers . Inhalable dust, endotoxin, fungal and bacterial contamination all exceeded levels found in similar vegetable fibre processing factories, since inhalable dust levels ranged from 10.4 to 79.8 mg/m(3) and inhalable bacterial levels between 4.7 and 190 x 10(6) cfu/m(3) . Soluble protein and endotoxin (r=0.99, P<0.0001), endotoxin and inhalable dust (r=0.94, P<0.005) and inhalable dust and protein (r=0.98, P<0.0001) were significantly correlated, suggesting that there was little variation in the composition of the dust from different sites or activities around the workplace . Andersen sampling gave an indication of background microbe levels, although no attempt was made to identify the specific microorganisms as all plates were significantly overgrown . Airborne assessments demonstrated that exposures were highly task specific . For example, sweeping the floor generated the highest exposure levels of total dust, protein, endotoxin, bacteria and fungi . Therefore, we have shown that a modern-day hemp fibre processing plant produces significant quantities of respirable dust which is highly contaminated with endotoxin and microorganisms . This organic dust has the potential to cause a range of ill health problems. J Anim Sci, 2001 Sep, 79(9), 2491 - 8 Rumen microbial sequestration of {2-(13)C}acetate in cattle; Kristensen NB; To investigate the impact of rumen microbial sequestration of VFA carbon on estimates of acetate availability based on intraruminal infusion of {2-(13)C} acetate, three nonlactating or low-yielding dairy cows were continuously intraruminally infused with {2-(13)C}acetate for 26 h . The 13C content of ruminal VFA, duodenal carbon, and fatty acids (FA) and AA isolated from liquid-associated ruminal microbes and duodenal DM was measured by an isotope ratio mass spectrometer interfaced to an elemental analyzer or a gas-liquid chromatograph . The ruminal gross production of acetate was 38 +/- 4 mol/d and could account for about 38% of the DE intake . Of the intraruminally infused 13C in {2-(13)C}acetate, 7.6 +/- 0.9% was recovered at the duodenum . The 13C content of ruminal propionate, butyrate, and valerate increased (P < 0.05) with intraruminal infusion of {2-(13)C}acetate . It was estimated that about 28% of the 13C intraruminally infused in {2-(13)C}acetate could be accounted for by duodenal 13C flow and absorption of non-acetate VFA . A number of FA isolated from liquid-associated ruminal microbes (C6, C12, C14, anteiso C15, and iso C15) were enriched with 13C (P < 0.05) at a level comparable to the enrichment of ruminal butyrate . Any absorption of these FA from the rumen would further contribute to non-acetate 13C uptake . A maximum of 72% of the ruminal gross production of acetate represented acetate absorption from the rumen in the present study . Consequently, previously used models using intraruminal isotope dilution techniques seem not to be appropriate for measuring acetate availability in ruminants . The number of metabolites exchanging carbon with acetate was found to be so high that assessments of the entire range of inter conversions seem to be practically impossible . Portal absorption studies are discussed as an alternative method of estimating VFA availability to the metabolism in ruminants. Can J Microbiol, 2001 Aug, 47(8), 773 - 6 Influence of the zinc hyperaccumulator Thlaspi caerulescens J . & C . Presl . and the nonmetal accumulator Trifolium pratense L . on soil microbial populations; Delorme TA et al.; Metal hyperaccumulator plants like Thlaspi caerulescens J . & C . Presl . are used for phytoremediation of contaminated soils . Since little is known about the rhizosphere of hyperaccumulators, the influence of T . caerulescens was compared with the effects of Trifolium pratense L . on soil microbes . High- and low-metal soils were collected near a zinc smelter in Palmerton, Penn . Soil pH was adjusted to 5.8 and 6.8 by the addition of Ca(OH)2 . Liming increased bacterial populations and decreased metal toxicity to levels allowing growth of both plants . The effects of the plants on total (culturable) bacteria, total fungi, as well as cadmium- and zinc-resistant populations were assessed in nonrhizosphere and rhizosphere soil . Both plants increased microbial populations in rhizosphere soil compared with nonrhizosphere soil . Microbial populations were higher in soils planted with T . pratense, but higher ratios of metal-resistant bacteria were found in the presence of T . caerulescens . We hypothesize that T . caerutescens acidifies its rhizosphere . Soil acidification in the rhizosphere of T . caerulescens would affect metal uptake by increasing available metals around the roots and consequently, increase the selection for metal-resistant bacteria . Soil acidification may be part of the hyperaccumulation process enhancing metal uptake from soil. Lakartidningen, 2001 Aug 22, 98(34), 3510 - 4 {Infection a dreaded central venous catheter-related complication . A reminder of the significance of good aseptics is justified}; Hammarskjold F et al.; The use of central venous catheters has increased markedly . Large numbers of patients are therefore at risk for catheter-related infections . This paper reviews the literature on prevention of intravascular catheter-related complications . Microbes colonising the catheter hubs and the skin around the insertion site are the source of most of these infections . By simple routines it is possible to reduce the risk for microbial spread from these sites to the bloodstream. Ann Allergy Asthma Immunol, 2001 Sep, 87(3), 181 - 8 A superantigen hypothesis for the pathogenesis of chronic hypertrophic rhinosinusitis, allergic fungal sinusitis, and related disorders; Schubert MS; BACKGROUND: Chronic eosinophilic-lymphocytic respiratory mucosal inflammatory disorders include hypertrophic sinus disease, allergic fungal sinusitis, allergic bronchopulmonary aspergillosis, and chronic severe asthma . They have many analogous or shared aspects of pathology at molecular, cellular, and clinical levels of analysis . OBJECTIVE: To propose a theory, and supporting data through comprehensive literature review, that unifies these diseases' pathogenesis . METHODS AND DATA SOURCES: Current medical literature was used as supportive background information . Reinterpretation of existing studies and reasoned speculation were used when necessary and identified where used . English language MEDLINE articles that referenced sinusitis, rhinosinusitis, allergic fungal sinusitis, asthma, allergic bronchopulmonary aspergillosis, nasal polyp, superantigen, and T cell receptor from 1983 to present were potentially used as background or supportive information . Additional referenced articles, published abstracts, and National Center for Biotechnology Information Entrez protein database searches were used . Case reports, studies, review articles, and textbooks were included . RESULTS: Multiple lines of evidence support the proposed hypothesis that microbial T cell superantigen production, persistence, and host-responsiveness are the fundamental components that unify the pathogenesis of all common chronic eosinophilic-lymphocytic respiratory mucosal inflammatory disorders . Superantigen amplification of preexisting immunopathology is the proposed mechanism for disease induction and maintenance . Preexisting immunopathology is created in the individual by a potential heterogeneity of immunopathologic signals that can include type I immediate hypersensitivity, other antigen-specific immune responses, cytokine dysregulation, eicosanoid dysregulation, various genetic mutations, and other molecular pathology . Although the ability to develop chronic severe inflammatory disease is dependent upon this immunopathology, host T cell receptor V beta genetics and persistent superantigen production/exposure at the respiratory mucosa by relevant superantigen-producing extra- or intracellular microbes are postulated to be required . This mechanism for disease pathogenesis may also apply to other disorders . Approaches to prove this theory and its predictions are presented . CONCLUSIONS: The pathogenesis of all the disorders discussed can be unified through the superantigen hypothesis proposed . Multiple lines of evidence support this hypothesis . How we view these common conditions will change, and new research into pathogenesis and treatment will occur if this proves true. Chir Organi Mov, 1999 Jul-Sep, 84(3), 247 - 51 Vancomycin prophylaxis in joint arthroplasty; Savarese A et al.; Postoperative infections in prosthetic surgery still constitute a serious problem, and one that is difficult to treat, because of the occurrence of agglomerates of microbes that are resistant to immune defenses and antibiotics . In nearly all cases, removal of the prosthesis is the only possible means of solving the problem of infection . The systematic use of antibiotic prophylaxis in surgery of this sort offers advantages in terms of a reduction in the risk of infection . The authors present a personal case series relative to the strategies of perioperative antibiotic prophylaxis used in cases of hip and knee arthroplasty; we refer to 233 joint arthroplasties performed between October 1993 and April 1996 . In all of the cases, perioperative prophylaxis with vancomycin chlorohydrate at a dose of 1 g i.v . 1 hour prior to surgery, and 6-8 hours after surgery was carried out . The choice of the antibiotic was based on the epidemiological knowledge of the literature and the experience on the ward. Science, 2001 Sep 21, 293(5538), 2248 - 51 Effects of size and temperature on metabolic rate; Gillooly JF et al.; We derive a general model, based on principles of biochemical kinetics and allometry, that characterizes the effects of temperature and body mass on metabolic rate . The model fits metabolic rates of microbes, ectotherms, endotherms (including those in hibernation), and plants in temperatures ranging from 0 degrees to 40 degrees C . Mass- and temperature-compensated resting metabolic rates of all organisms are similar: The lowest (for unicellular organisms and plants) is separated from the highest (for endothermic vertebrates) by a factor of about 20 . Temperature and body size are primary determinants of biological time and ecological roles. Curr Rheumatol Rep, 2001 Oct, 3(5), 428 - 34 Insights into the Role of Infection in the Spondyloarthropathies; Liu Y et al.; Certain infections play an important role in the pathogenesis of the human leukocyte antigen (HLA)-B27-associated reactive arthritis . Whether infections play a role in other forms of spondyloarthropathies is not as clear . The role of HLA-B27 as an antigen-presenting molecule is important in the pathogenesis of these diseases . Recent evidence has been obtained indicating that this molecule may have other functions unrelated to antigen-presentation in the interaction of reactive arthritis-triggering microbes and host . This paper reviews the recent studies on the role of infection in the spondyloarthropathies. Int Immunopharmacol, 2001 Sep, 1(9-10), 1651 - 67 Inflammation, carcinogenesis and cancer; Fitzpatrick FA; To fulfill their role in host-defense, granulocytes secrete chemically reactive oxidants, radicals, and electrophilic mediators . While this is an effective way to eradicate pathogenic microbes or parasites, it inevitably exposes epithelium and connective tissue to certain endogenous genotoxic agents . In ordinary circumstances, cells have adequate mechanisms to reduce the genotoxic burden imposed by these agents to a negligible level . However, inflammation persisting for a decade eventually elevates the risk of cancer sufficiently that it is discernible in case control epidemiological studies . Advances in our understanding of tumor suppressors and inflammatory mediators offer an opportunity to assess the molecular and cellular models used to guide laboratory investigations of this phenomenon . Disappointing results from recent clinical trials with anti-oxidant interventions raise questions about the risks from specific endogenous agents such as hydrogen peroxide and oxy radicals . Simultaneously, the results from the anti-oxidant trials draw attention to an alternate hypothesis, favoring epigenetic inactivation of key tumor suppressors, such as p53, and the consequent liability this places on genomic integrity. Virchows Arch, 2001 Aug, 439(2), 152 - 7 Identification of syt-ssx fusion transcripts in both epithelial and spindle cell components of biphasic synovial sarcoma in small tissue samples isolated by membrane-based laser microdissection; Nishio J et al.; In order to confirm the presence of SYT-SSX fusion gene in epithelial and spindle cell components of synovial sarcoma, we performed a nested reverse transcriptase-polymerase chain reaction (RT-PCR) using microbeam microdissection of membrane-mounted native tissue (MOMeNT) technique applied on formalin-fixed, paraffin-embedded tumor specimens from two biphasic synovial sarcomas and a control tissue of adamantinoma . Small targeted portions of either an epithelial or spindle cell component of the tumor tissue were microdissected together with the supporter membrane, by using an ultraviolet (337-nm) pulsed laser microbeam coupled into a robot-stage microscope with infinity optics . The SYT-SSX fusion transcript was detected in epithelial and spindle cell components of both biphasic synovial sarcomas, but not in the control tissue . Southern blot analysis also confirmed that the detected messages were derived from the SYT-SSX fusion gene . In conclusion, the microbeam MOMeNT is a useful method for isolating selected small portions from tissue sections . The SYT-SSX fusion gene is present in both cellular components of biphasic synovial sarcoma and is involved in oncogenesis of the synovial sarcoma rather than in morphologic epithelial differentiation . Therefore, in spite of the variable proportions of each component, our results confirm that the synovial sarcoma is of monoclonal origin. J Exp Med, 2001 Sep 17, 194(6), 863 - 9 Subsets of human dendritic cell precursors express different toll-like receptors and respond to different microbial antigens; Kadowaki N et al.; Toll-like receptors (TLRs) are ancient microbial pattern recognition receptors highly conserved from Drosophila to humans . To investigate if subsets of human dendritic cell precursors (pre-DC), including monocytes (pre-DC1), plasmacytoid DC precursors (pre-DC2), and CD11c(+) immature DCs (imDCs) are developed to recognize different microbes or microbial antigens, we studied their TLR expression and responses to microbial antigens . We demonstrate that whereas monocytes preferentially express TLR 1, 2, 4, 5, and 8, plasmacytoid pre-DC strongly express TLR 7 and 9 . In accordance with these TLR expression profiles, monocytes respond to the known microbial ligands for TLR2 (peptidoglycan {PGN}, lipoteichoic acid) and TLR4 (lipopolysaccharide), by producing tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 . In contrast, plasmacytoid pre-DCs only respond to the microbial TLR9-ligand, CpG-ODNs (oligodeoxynucleotides {ODNs} containing unmethylated CpG motifs), by producing IFN-alpha . CD11c(+) imDCs preferentially express TLR 1, 2, and 3 and respond to TLR 2-ligand PGN by producing large amounts of TNF-alpha, and to viral double-stranded RNA-like molecule poly I:C, by producing IFN-alpha and IL-12 . The expression of distinct sets of TLRs and the corresponding difference in reactivity to microbial molecules among subsets of pre-DCs and imDCs support the concept that they have developed through distinct evolutionary pathways to recognize different microbial antigens. Curr Infect Dis Rep, 2001 Oct, 3(5), 419 - 426 Genetic Analysis of Host Responses in Sepsis; Beutler B et al.; During much of the past century, the microbe itself stood at the heart of microbial pathogenesis . Little thought was devoted to the host per se, though it was granted that differences in susceptibility to certain infections did exist between individuals, and between different ethnic groups . During the past 20 years, extraordinary strides in our grasp of mammalian genetics have made the host side of the equation far more approachable . A restricted collection of genes now presents itself as the likely repository for genetic differences that foretell susceptibility to infectious disease . The Toll-like receptors, of which 10 are presently known to exist in humans, offer an excellent example of this genetic reductionism, in that they embody the afferent component of the innate immune system, and strongly influence the containment of an infection from its earliest stages . The Toll-like receptors were identified as the culmination of a long and relentless inquiry into the yet-unsolved clinical problem of sepsis. Phytochemistry, 2001 Oct, 58(3), 395 - 401 Production of loline alkaloids by the grass endophyte, Neotyphodium uncinatum, in defined media; Blankenship JD et al.; Lolines (saturated 1-aminopyrrolizidines with an oxygen bridge) are insecticidal alkaloids produced in symbioses of certain Epichloe (anamorph-Neotyphodium) species (fungal endophytes) with grasses, particularly of the genera Lolium and Festuca . Prior to the present study, it was unknown whether lolines were of plant or fungal origin . Neotyphodium uncinatum, the common endophyte of meadow fescue (Lolium pratense=Festuca pratensis) produced loline, N-acetylnorloline, and N-formylloline when grown in the defined minimal media at pH 5.0-7.5, with both organic and inorganic nitrogen sources and sugars as carbon sources . In contrast, lolines were not detected in complex medium cultures . GC-MS and 13C NMR spectroscopic analyses confirmed the identity of the alkaloids isolated from the defined medium cultures . Lolines accumulated to ca . 700 mg/l (4 mM) in cultures with 16.7 mM sucrose and 15-30 mM asparagine, ornithine or urea . Kinetics of loline production and fungal growth were assessed in defined medium with 16.7 mM sucrose and 30 mM ornithine . The alkaloid production rate peaked after the onset of stationary phase, as is common for secondary metabolism in other microbes. Curr Opin Rheumatol, 2001 Jul, 13(4), 300 - 4 Infectious agents in chronic rheumatic diseases; Hyrich KL et al.; The possible role of infectious microorganisms in the disease process of both arthritis and autoimmunity continue to attract both basic and clinical researchers . However, proving a causal role for these suspects is a very difficult and complicated task . This article provides an update on various mechanisms in which microbes may play roles as inciting or perpetuating factors in the pathogenesis of connective tissue disease . It also focuses on current theories that specific microorganisms may play a role in rheumatoid arthritis and ankylosing spondylitis. Birth, 2001 Sep, 28(3), 161 - 5 Does bathing newborns remove potentially harmful pathogens from the skin? Medves JM, O'Brien B. BACKGROUND: Newborn infants are routinely bathed after birth partly to reduce the possibility of transmitting potential pathogens to others . The extent to which a mild soap reduces the quantity and type of microbes found on the skin through normal colonization has not been reported . The objective of the study was to compare colonization rates between infants bathed in soap and water and infants bathed in plain water . METHOD: One hundred and forty infants were randomly assigned to one group bathed in a mild pH neutral soap and water or to another group bathed in water alone . Microbiology swabs were taken on three occasions (before the first bath, 1 hour after the bath, and 24 hours after birth) from two sites (anterior fontanelle and umbilical area) . RESULTS: No difference occurred between groups on type or quantity of organisms found at each time period . Skin colonization is a function of time, and the quantity of organisms identified increased over time (Friedman A 2 = 111.379, df = 5, p < 0.001) . CONCLUSIONS: Bathing with mild soap as opposed to bathing in water alone has minimal effect on skin bacterial colonization . Skin colonization increased over time . The findings did not support the efficacy of bathing with soap and water to reduce skin colonization of bacterial pathogens . Although the incidence of potential pathogens colonizing the skin during the first day of life is low and unlikely to pose a risk to healthy newborns, health care professionals may wish to wear gloves until the infant has been bathed. Adv Drug Deliv Rev, 2001 Jul 28, 49(3), 301 - 15 Caveolae in the uptake and targeting of infectious agents and secreted toxins; Norkin LC; A variety of microbial pathogens, including viruses, intracellular bacteria, and prions, as well as certain secreted bacterial toxins, can now be added to the list of ligands that enter cells via caveolae or caveolae-like membrane domains . In general, the caveolae-mediated entry pathway results in transport of these microbes and toxins to intracellular destinations that are different from that of cargo entering by other means . As a result, the caveolae-mediated entry pathway can profoundly affect the host cell-pathogen interaction long after entry has occurred . Furthermore, some microbes such as SV40 that enter via cavolae will be valuable as probes to analyze certain poorly understood intracellular trafficking pathways, such as retrograde transport to the ER . Also, viruses that enter via caveolae may have unique potential as gene and drug delivery vectors . In addition, some extracellular microbial pathogens, such as Pneumocystis carinii, may also interact with host cells via caveolae . Finally, caveolae may play a role in host immune defense mechanisms. Methods Cell Sci, 2000, 22(4), 329 - 41 A method to purify bacteria-containing phagosomes from infected macrophages; Luhrmann A et al.; When small particles, such as microorganisms, are taken up by macrophages, they are wrapped with a portion of the host cell plasma membrane and ingested, creating a new organelle, the phagosome . This phagosome matures stepwise as newly formed endosomes do, finally forming a phagolysosome, a process that contributes to killing of ingested microbes and to the presentation of microbial antigens on the surface of the phagocyte . Some pathogenic bacteria, however, reprogram the phagocytic cell in such a way that the phagosome will either be arrested in an early stage of maturation or will be diverted and create an unusual, novel phagosomal compartment . To study the molecular processes that underly biogenesis of bacteria-containing phagosomes, we have established a method to isolate and to biochemically analyse bacteria- containing phagosomes . This method consists of mechanical lysis of infected macrophages, production of a postnuclear supernatant followed by fractionation in a discontinuous sucrose density gradient, separation through a Ficoll cushion, and by a final concentration step . These phagosome preparations contain very little endosomal or lysosomal contamination (the organelles of most concern when studying phagosome biogenesis) and very little Golgi- and plasma membrane-derived contamination, but do contain some mitochondrial and ER contamination . This method could also be used to study bacterial factors (proteins, RNA) produced while in phagosomes. Cell Transplant, 2001, 10(4-5), 403 - 8 Functional comparison of the single-layer agarose microbeads and the developed three-layer agarose microbeads as the bioartificial pancreas: an in vitro study; Xu B et al.; In this study, the insulin secretory characteristics of the microencapsulated hamster islets were studied during long-term culture . The hamster islets were encapsulated as single-layer agarose microbeads or three-layer agarose microbeads with agarose and agarose containing poly(styrene sulfonic acid) (PSSa), respectively . The influence of PSSa on the function of the rat islets microencapsulted in three-layer microbeads was primarily monitored . The aim of this study was to examine the influence of the PSSa on the in vitro function of the islets encapsulated in the agarose/PSSa microbeads compared with single-layer agarose microbeads during long-term culture . The microbeads were cultured for 30 days in medium of Eagle's MEM at 37 degrees C in 5% CO2 and 95% air . The basal insulin secretion into the culture medium was measured daily during the first 12 days and two times per week until 30 days . The microbeads were subjected to static incubation test on the 10th, 20th, and 30th day during culture . The basal insulin secretion level of the agarose/PSSa microbeads was significantly higher than that of single-layer agarose microbeads . The static incubation tests revealed a similar pattern of insulin secretion from both microbeads when they were exposed to high glucose challenge . In the static incubation test, both could significantly increase insulin release to more than 6.61 times (stimulation index) in response to high glucose stimulation and could significantly decrease when glucose concentration returned from high glucose to low glucose on the 10th, 20th, and 30th day of culture . This study demonstrated that the hamster islets enclosed in agarose/PSSa hydrogel not only continuously secreted basal amounts of insulin, but also maintained their response to high glucose stimulation similar to the agarose microbeads . The above results together with those of our previous in vivo study suggest that the three-layer microbeads (agarose/PSSa) are well suitable for xenotransplantation of islets for the clinical application. Appl Microbiol Biotechnol, 2001 Aug, 56(3-4), 326 - 38 Microbial xylanases and their industrial applications: a review; Beg QK et al.; Despite an increased knowledge of microbial xylanolytic systems in the past few years, further studies are required to achieve a complete understanding of the mechanism of xylan degradation by microorganisms and their enzymes . The enzyme system used by microbes for the metabolism of xylan is the most important tool for investigating the use of the second most abundant polysaccharide (xylan) in nature . Recent studies on microbial xylanolytic systems have generally focussed on induction of enzyme production under different conditions, purification, characterization, molecular cloning and expression, and use of enzyme predominantly for pulp bleaching . Rationale approaches to achieve these goals require a detailed knowledge of the regulatory mechanism governing enzyme production . This review will focus on complex xylan structure and the microbial enzyme complex involved in its complete breakdown, studies on xylanase regulation and production and their potential industrial applications, with special reference to biobleaching. Annu Rev Microbiol, 2001, 55, 357 - 80 A community of ants, fungi, and bacteria: a multilateral approach to studying symbiosis; Currie CR; The ancient and highly evolved mutualism between fungus-growing ants and their fungi is a textbook example of symbiosis . The ants carefully tend the fungus, which serves as their main food source, and traditionally are believed to be so successful at fungal cultivation that they are able to maintain the fungus free of microbial pathogens . This assumption is surprising in light of theories on the evolution of parasitism, especially for those species of ants that have been clonally propagating their cultivars for millions of years . Recent work has established that, as theoretically predicted, the gardens of fungus-growing ants are host to a specialized, virulent, and highly evolved fungal pathogen in the genus Escovopsis . In addition, the ants have evolved a mutualistic association with filamentous bacteria (actinomycetes) that produce antibiotics that suppress the growth of Escovopsis . Thus, the attine symbiosis appears to be a coevolutionary "arms race" between the garden parasite Escovopsis on the one hand and the ant-fungus-actinomycete tripartite mutualism on the other . These recent findings indicate that microbes may be key components in the regulation of other symbiotic associations between higher organisms. J Sediment Res A Sediment Petrol Process, 2000 May, 70(3), 565 - 85 Depositional facies and aqueous-solid geochemistry of travertine-depositing hot springs (Angel Terrace, Mammoth Hot Springs, Yellowstone National Park, U.S.A.); Fouke BW et al.; Petrographic and geochemical analyses of travertine-depositing hot springs at Angel Terrace, Mammoth Hot Springs, Yellowstone National Park, have been used to define five depositional facies along the spring drainage system . Spring waters are expelled in the vent facies at 71 to 73 degrees C and precipitate mounded travertine composed of aragonite needle botryoids . The apron and channel facies (43-72 degrees C) is floored by hollow tubes composed of aragonite needle botryoids that encrust sulfide-oxidizing Aquificales bacteria . The travertine of the pond facies (30-62 degrees C) varies in composition from aragonite needle shrubs formed at higher temperatures to ridged networks of calcite and aragonite at lower temperatures . Calcite "ice sheets", calcified bubbles, and aggregates of aragonite needles ("fuzzy dumbbells") precipitate at the air-water interface and settle to pond floors . The proximal-slope facies (28-54 degrees C), which forms the margins of terracette pools, is composed of arcuate aragonite needle shrubs that create small microterracettes on the steep slope face . Finally, the distal-slope facies (28-30 degrees C) is composed of calcite spherules and calcite "feather" crystals . Despite the presence of abundant microbial mat communities and their observed role in providing substrates for mineralization, the compositions of spring-water and travertine predominantly reflect abiotic physical and chemical processes . Vigorous CO2 degassing causes a +2 unit increase in spring water pH, as well as Rayleigh-type covariations between the concentration of dissolved inorganic carbon and corresponding delta 13C . Travertine delta 13C and delta 18O are nearly equivalent to aragonite and calcite equilibrium values calculated from spring water in the higher-temperature (approximately 50-73 degrees C) depositional facies . Conversely, travertine precipitating in the lower-temperature (< approximately 50 degrees C) depositional facies exhibits delta 13C and delta 18O values that are as much as 4% less than predicted equilibrium values . This isotopic shift may record microbial respiration as well as downstream transport of travertine crystals . Despite the production of H2S and the abundance of sulfide oxidizing microbes, preliminary delta 34S data do not uniquely define the microbial metabolic pathways present in the spring system . This suggests that the high extent of CO2 degassing and large open-system solute reservoir in these thermal systems overwhelm biological controls on travertine crystal chemistry. Icarus, 2000 Aug, 146(2), 343 - 59 The ultraviolet environment of Mars: biological implications past, present, and future; Cockell CS et al.; A radiative transfer model is used to quantitatively investigate aspects of the martian ultraviolet radiation environment,