Microorganism

Chemosphere, 2004 Feb, 54(7), 823 - 9
Effect of Fe0 quantity on the efficiency of integrated microbial-Fe0 treatment processes; Fernandez-Sanchez JM et al.; Batch experiments were conducted with different reaction systems to investigate how the treatment efficiency of integrated microbial-Fe0 processes is affected by the amount of Fe0 added . Abiotic experiments with hexavalent chromium and carbon tetrachloride mixtures corroborated that different pollutants could compete for reactive sites on the iron surface, which would hinder specific degradation rates when the available Fe0 surface area is relatively small (e.g., 11 m(2) l(-1)) . In such cases, reductive precipitation of chromium could occlude reactive sites and significantly inhibit removal efficiency . Microbial participation in the cleanup process was also influenced by the amount of Fe0 added . Increasing the Fe0 dose (and thus the available surface area) had a stimulatory effect possibly due to a higher production of cathodic H2, which can be used as electron donor for reductive biotransformation of many pollutants . However, high Fe0 doses had an inhibitory effect due to a corrosion-induced increase in pH beyond the optimum range of the bacteria . This suggest that there may be a system-specific, optimum quantity of Fe0 that satisfies availability requirements to preclude contaminant competition for reactive sites and biological requirements for H2 production while minimizing inhibitory increases in pH . Results also confirmed extensive RDX mineralization in bioaugmented (but not in abiotic) Fe0 systems, and support the notion that permeable reactive iron barriers performance might be enhanced by the participation of some microorganisms.

Scand J Immunol, 2003 Dec, 58(6), 642 - 8
Development of immunoglobulin A in infancy and childhood; Weemaes C et al.; Serum and salivary concentrations of immunoglobulin A1 (IgA1) and IgA2 were studied in 105 Icelandic children aged 0-12 years . Serum concentrations of both IgA1 and IgA2 increased slightly (P < 0.001) during childhood . The salivary IgA1/IgA2 ratio tended to decrease during the same period; this trend is less apparent when omitting the youngest children . The salivary IgA1 and IgA2 output could be high, even in children with low levels of serum IgA . Only polymeric IgA was found in whole saliva . Interestingly, in serum, most IgA1 and IgA2 were polymeric during infancy . The proportion of polymeric IgA decreased, when the concentration of IgA increased . The polymeric form of IgA might provide the infant with better protection against invading microorganisms by activation of the innate immune mechanisms.

Br J Dermatol, 2003 Nov, 149(5), 1029 - 34
Effect of Mycobacterium vaccae on atopic dermatitis in children of different ages; Arkwright PD et al.; BACKGROUND: Exposure to environmental microorganisms is associated with variations in the prevalence and severity of atopic diseases . We have previously shown that administration of a Mycobacterium vaccae suspension significantly reduced the severity of atopic dermatitis (AD) in children aged 5-18 years . OBJECTIVES: This study aimed to extend these observations to younger children . METHODS: Fifty-six children aged 2-6 years with moderate to severe AD were enrolled in a randomized, double-blind, placebo-controlled trial and given one intradermal injection of either killed M . vaccae suspension or buffer solution (placebo) . Skin surface area affected and dermatitis severity score were assessed before and 1, 3 and 6 months after treatment . RESULTS: Although a 38-54% reduction in surface area affected by dermatitis was noted at all time points after M . vaccae administration (P = 0.005), this improvement was not significantly different from that observed in the placebo group . Meta-analysis of this and our previous cohort (97 children aged 2-18 years) showed that M . vaccae was associated with a significant improvement in clinical severity at all ages, whereas within the placebo group, younger but not older children showed a similar improvement . CONCLUSIONS: Despite a reduction in clinical severity associated with M . vaccae at all ages, no benefit could be found after administering M . vaccae to children with AD aged 2-6 years when compared with placebo . M . vaccae may offer greater benefit in children over 5 years old, whose AD appears less likely to regress spontaneously.

Res Commun Mol Pathol Pharmacol, 2002, 111(1-4), 103 - 13
Inhibitory effect of eight simple coumarins on the lucigenin enhanced chemiluminescence of rabbit neutrophils; Kabeya LM et al.; The generation of superoxide anion (O2*-) and other reactive oxygen species (ROS), such as HO*, HOCl, NO, 1O2 and H2O2, by stimulated polymorphonuclear leukocytes during phagocytosis is a major mechanism of host defense against invading microorganisms . However, large amounts of ROS are suggested to be responsible for many diseases . In this work we studied the inhibitory effect of eight simple coumarins on O2*- generation by rabbit neutrophil upon stimulation with opsonized zymosan, using lucigenin enhanced chemiluminescence assay . We observed that coumarins containing hydroxy or acetoxy substituents at position 7 of the benzopyrone ring were the most active ones (IC50 values ranging from 6.0 +/- 2.8 to 18.6 +/- 2.6 micromol/L) . Substitution of these groups by allyloxy or metoxy groups decreased the activity and unsubstituted coumarin had no effect . Cell damage after exposure to 200 micromol/L of each compound was determined by measurement of the activity of the released cytosolic lactate dehydrogenase and by Trypan Blue dye exclusion test . None of the drugs affected significantly the cellular viability.

Eye, 2003 Nov, 17(8), 949 - 56
Immune defense at the ocular surface; Akpek EK et al.; The ocular surface is constantly exposed to a wide array of microorganisms . The ability of the outer ocular system to recognize pathogens as foreign and eliminate them is critical to retain corneal transparency, hence preservation of sight . Therefore, a combination of mechanical, anatomical, and immunological defense mechanisms has evolved to protect the outer eye . These host defense mechanisms are classified as either a native, nonspecific defense or a specifically acquired immunological defense requiring previous exposure to an antigen and the development of specific immunity . Sight-threatening immunopathology with autologous cell damage also can take place after these reactions . This article discusses the innate and acquired corneal elements of the immune defense at the ocular surface . The relative roles of the various factors contributing to prevention of eye infection remain to be fully defined.

Plant Physiol, 2003 Dec, 133(4), 1968 - 78 Epub 2003 Nov 20.
Identification of AtNDI1, an internal non-phosphorylating NAD(P)H dehydrogenase in Arabidopsis mitochondria; Moore CS et al.; Plant mitochondria contain non-phosphorylating NAD(P)H dehydrogenases (DHs) that are not found in animal mitochondria . The physiological function, substrate specificity, and location of enzymes within this family have yet to be conclusively determined . We have linked genome sequence information to protein and biochemical data to identify that At1g07180 (SwissProt Q8GWA1) from the Arabidopsis Genome Initiative database encodes AtNDI1, an internal NAD(P)H DH in Arabidopsis mitochondria . Three lines of evidence are presented: (a) . The predicted protein sequence of AtNDI1 has high homology with other designated NAD(P)H DHs from microorganisms, (b) . the capacity for matrix NAD(P)H oxidation via the rotenone-insensitive pathway is significantly reduced in the Atndi1 mutant plant line, and (c) . the in vitro translation product of AtNDI1 is imported into isolated mitochondria and located on the inside of the inner membrane.

Biochem Biophys Res Commun, 2003 Dec 5, 312(1), 205 - 8
Reappraisal of the 20th-century version of amino acid metabolism; Katagiri M et al.; In this article, we advocate the radical revision of the 20th-century version of amino acid metabolism as follows . (1) Classic studies on the incorporation of {15N}ammonia into glutamate, once considered to be an epoch-making event, are not distinctive proof of the ability of animals to utilize ammonia for the synthesis of alpha-amino nitrogen . (2) Mammalian glutamate dehydrogenase has been implicated to function as a glutamate-synthesizing enzyme albeit lack of convincing proof . This enzyme, in combination with aminotransferases, is now known to play an exclusive role in the metabolic removal of amino nitrogen and energy production from excess amino acids . (3) Dr . William C Rose's "nutritionally nonessential amino acids" are, of course, essential in cellular metabolism; the nutritional nonessentiality is related to their carbon skeletons, many of which are intermediates of glycolysis or the TCA cycle . Obviously, the prime importance of amino acid nutrition should be the means of obtaining amino nitrogen . (4) Because there is no evidence of the presence of any glutamate-synthesizing enzymes in mammalian tissues, animals must depend on plants and microorganisms for preformed alpha-amino nitrogen . This is analogous to the case of carbohydrates . (5) In contrast, individual essential amino acids, similar to vitamins and essential fatty acids, should be considered important nutrients that must be included regularly in sufficient amounts in the diet.

Am J Reprod Immunol, 2003 Sep, 50(3), 254 - 62
Placental lesions caused by experimental infection of Sprague-Dawley rats with Mycoplasma pulmonis; Peltier MR et al.; PROBLEM: Sprague-Dawley (SD) rats infected during pregnancy with Mycoplasma pulmonis display adverse pregnancy outcomes that are similar to those observed in women with chorioamnionitis and may provide a good model system for this disease . The placental lesions caused by this microorganism, however, have not been thoroughly characterized . METHOD OF STUDY: Rats were infected with 10(7) colony-forming units (CFU) M . pulmonis or vehicle control on gestation day (gd) 14 and were euthanized on gd 16-18 . Tissues were fixed in 10% neutral buffered formalin, embedded in paraffin, sectioned at 4 microm, and stained with hematoxylin and eosin (H & E) . The slides were coded and examined by a blinded pathologist using light microscopy . RESULTS: Infection with M . pulmonis was associated with necrosis of trophoblast giant cells at gd 18 . Significantly more neutrophils were observed in the decidual region of the apex of the placenta in M . pulmonis infected animals . The vast majority of neutrophils, however, were observed in the decidua in the lateral regions of the placenta and in the adjacent endometrium . CONCLUSIONS: Infection of SD rats with M . pulmonis resulted in histological placentitis similar to that described in deciduitis of humans and represents a good model system for investigations into the pathophysiology of intrauterine infection . The influx of neutrophils seems to migrate from the endometrium towards the lateral regions of the placenta near Reichert's membrane and the divergence of the parietal yolk sac.

Biotechniques, 2003 Nov, 35(5), 996 - 1002, 1004-5
Nitrogenase genes in PCR and RT-PCR reagents: implications for studies of diversity of functional genes; Zehr JP et al.; Studies of the diversity of microorganisms in the environment have been facilitated by use of PCR and reverse transcription PCR (RT-PCR) . Inhibition of the PCR by complex sample matrices and low abundance of some target microorganisms require the use of high-sensitivity amplification procedures, involving a large number of cycles or nested PCR methods . Using these methods, we frequently observed contamination of the amplification reagents, including polymerases, by genomic DNA containing nitrogenase (nifH) and rRNA genes . Contaminating genes were sequenced and found to belong to a variety of rRNA clades, but only three major nifH clades . These sequence types included a few nifH sequences reported in previous studies of the environment . Contamination could be reduced by restriction digestion and ultrafiltration of PCR reagents, but efficiency of amplification was also reduced . Our results suggest that studies relying on large numbers of PCR amplification cycles to assess environmental gene diversity should take precautions to assure that clone libraries generated from amplified PCR products are not the result of contaminated PCR reagents.

Morfologiia, 2003, 124(4), 66 - 9
{Functional significance of "blind" outgrowths of the lymphatic capillary network}; Aminova GG; The work was aimed at the study of the structural peculiarities of "blind" outgrowths of the lymphatic capillaries and to define their role in the pathogenesis of some diseases . In experiments performed in rabbits by studying the resorption of colloidal solutions, suspensions and hen's blood the subsidiary functional significance of the portion of "blind" outgrowths of the lymphatic capillary network of tendinous center of diaphragm was established . The clavate outgrowths with the narrow mouth may accumulate and store the exogenous material in their lumen . It is concluded that they may accumulate the microorganisms that provoke the relapses of diseases.

Bioelectromagnetics, 2003, Suppl 6, S74 - 100
Radio frequency electromagnetic fields: cancer, mutagenesis, and genotoxicity; Heynick LN et al.; We present critiques of epidemiologic studies and experimental investigations, published mostly in peer-reviewed journals, on cancer and related effects from exposure to nonionizing electromagnetic fields in the nominal frequency range of 3 kHz to 300 GHz of interest to Subcommittee 4 (SC4) of the International Committee on Electromagnetic Safety (ICES) . The major topics discussed are presented under the headings Epidemiologic and Other Findings on Human Exposure, Mammals Exposed In Vivo, Mammalian Live Tissues and Cell Preparations Exposed In Vitro, and Mutagenesis and Genotoxicity in Microorganisms and Fruit Flies . Under each major topic, we present minireviews of papers on various specific endpoints investigated . The section on Epidemiologic and Other Findings on Human Exposure is divided into two subsections, the first on possible carcinogenic effects of exposure from emitters not in physical contact with the populations studied, for example, transmitting antennas and other devices . Discussed in the second subsection are studies of postulated carcinogenic effects from use of mobile phones, with prominence given to brain tumors from use of cellular and cordless telephones in direct physical contact with an ear of each subject . In both subsections, some investigations yielded positive findings, others had negative findings, including papers directed toward experimentally verifying positive findings, and both were reported in a few instances . Further research on various important aspects may resolve such differences . Overall, however, the preponderance of published epidemiologic and experimental findings do not support the supposition that in vivo or in vitro exposures to such fields are carcinogenic . Published 2003 Wiley-Liss, Inc.

Ann Acad Med Singapore, 2003 Sep, 32(5), 710 - 4
From genesis to genes; Tan SK; Since the beginning of time, our ancestors have been plagued by illnesses and injuries that are not too different from today's diseases . Evidence from prehistoric times and ancient civilisations have shown man's attempts at trying to understand the nature and treatment of these conditions . It was not till the early 19th century that the scientific basis of modern medicine was firmly established when microorganisms were discovered and found to be the cause of many of these illnesses . The 20th century saw quantum leaps made in the understanding of the function of the human body and the therapeutic measures aimed at restoration of any such malfunction . The end of the last millennium was marked by historic achievements made in the Life Sciences, in particular the completion of the sequencing of the Human Genome--the code of life . The beginning of the 21st century has already seen many breakthroughs in medical sciences, especially in the fields of stem cell technology and gene therapy . The number of known illnesses directly related to genetic defects or abnormalities have increased exponentially . Many of today's scourges can be prevented or more effectively treated . Our ability to utilise this new knowledge to combat the ravages of the ageing process and its associated illnesses--degenerative diseases and cancers offer much hope for the future.

Appl Microbiol Biotechnol, 2004 May, 64(4), 576 - 87 Epub 2003 Nov 18.
Intrinsic bioremediability of an aromatic hydrocarbon-polluted groundwater: diversity of bacterial population and toluene monoxygenase genes; Cavalca L et al.; The functional and phylogenetic biodiversity of bacterial communities in a benzene, toluene, ethylbenzene and xylene (BTEX)-polluted groundwater was analysed . To evaluate the feasibility of using an air sparging treatment to enhance bacterial degradative capabilities, the presence of degrading microorganisms was monitored . The amplification of gene fragments corresponding to toluene monooxygenase (tmo), catechol 1,2-dioxygenase, catechol 2,3-dioxygenase and toluene dioxygenase genes in DNA extracted directly from the groundwater samples was associated with the presence of indigenous degrading bacteria . Five months of air injection reduced species diversity in the cultivable community (as calculated by the Shannon-Weaver index), while little change was noted in the degree of biodiversity in the total bacterial community, as characterised by denaturing gradient gel electrophoresis (DGGE) analysis . BTEX-degrading strains belonged to the genera Pseudomonas, Microbacterium, Azoarcus, Mycobacterium and Bradyrhizobium . The degrading capacities of three strains in batch liquid cultures were also studied . In some of these microorganisms different pathways for toluene degradation seemed to operate simultaneously . Pseudomonas strains of the P24 operational taxonomic unit, able to grow only on catechol and not on BTEX, were the most abundant, and were present in the groundwater community at all stages of treatment, as evidenced both by cultivation approaches and by DGGE profiles . The presence of different tmo-like genes in phylogenetically distant strains of Pseudomonas, Mycobacterium and Bradyrhizobium suggested recent horizontal gene transfer in the groundwater .

Chem Phys Lipids, 2003 Dec, 126(2), 225 - 32
Archaeal lipids forming a low energy-surface on air-water interface; Kitano T et al.; Archaea or archaebacteria are the microorganism living in extreme environments such as hot springs and salt lakes . The membrane is featured universally by lipids which possess saturated polyisoprenoid chains in the hydrophobic moiety . This paper concerns the surface properties of Langmuir membranes made of archaeal lipid models (AL) bearing a phytanyl group or (3RS, 7R, 11R)-3,7,11,15-tetramethylhexadecyl group . All of the AL provide a Langmuir membrane on an air-water interface with an abnormally low surface tension (32-37 mN/m at 20-70 degrees C), while the conventional lipids having n-alkyl chains give membranes of 54-56 mN/m . The abnormally low energy surface of AL lipids is considered to arise from the bulky and fluid polyisoprenoid chain.

Biochem Biophys Res Commun, 2003 Nov 21, 311(3), 597 - 603
Functionally important amino acids in Saccharomyces cerevisiae aspartate kinase; Bareich DC et al.; Saccharomyces cerevisiae aspartate kinase (AK(Sc)) phosphorylates L-Asp as the first step in the aspartate pathway responsible for the biosynthesis of L-Thr, L-Met, and L-Ile in microorganisms and plants . Using site-directed mutagenesis, we have evaluated the importance of residues in AK(Sc) that are strongly conserved among aspartate kinases or in other small molecule kinases . Steady state kinetic analysis of the purified AK(Sc) variants reveals that several of the targeted amino acids, particularly K18 and H292, have important roles in the enzymatic reaction . These results provide the first identification of amino acid residues crucial to the action of this important metabolic enzyme.

Oral Microbiol Immunol, 2003 Dec, 18(6), 339 - 49
Induction of T-cell apoptosis by Actinobacillus actinomycetemcomitans mutants with deletion of ltxA and cdtABC genes: possible activity of GroEL-like molecule; Nalbant A et al.; The pathogenic bacterium Actinobacillus actinomycetemcomitans expresses a leukotoxin (Ltx) and cytolethal distending toxin (CDT) with cytolytic properties . CDT also has cytostatic properties, inducing a G2 cell cycle block . The extent of the contribution of these, as well as other toxins, to the cytolytic and cytostatic activities of this microorganism have not been defined and the aim of this study was to determine their contribution . To that end, a naturally transformable A . actinomycetemcomitans clinical strain (D7S-smooth) was used to construct a series of deletion mutants (DeltacdtA, DeltacdtB, DeltacdtC, DeltacdtABC, DeltaltxA, DeltaltxA/DeltacdtABC) . Human peripheral blood mononuclear cells were incubated with cell-associated and extracellular bacterial preparations . The ability of wild type and isogenic mutants to induce T-cell apoptosis and cell cycle arrest was compared . The expression of ltxA and each of the cdt gene loci partially contributed to A . actinomycetemcomitans apoptosis, since each of the isogenic mutants exhibited reduced ability to induce T-cell apoptosis . Conversely, the ability to induce cell cycle block was abolished in each of the cdt isogenic mutants . A mutant with simultaneous deletion of ltxA and cdtABC genes retained potent ability to induce apoptosis in its cell-associated, but not extracellular, preparation . Neutralization with Escherichia coli anti-GroEL monoclonal antibody, lead to significant diminution of apoptosis-inducing activity of the DeltaltxA/DeltacdtABC cell-associated preparation . These data provide evidence for the expression of other A . actinomycetemcomitans cytolytic molecule(s) distinct from CDT and leukotoxin, with a possible role for GroEL-like molecule in T-cell apoptosis.

Eur J Biochem, 2003 Nov, 270(21), 4243 - 53
Reconstruction of the central carbon metabolism of Aspergillus niger; David H et al.; The topology of central carbon metabolism of Aspergillus niger was identified and the metabolic network reconstructed, by integrating genomic, biochemical and physiological information available for this microorganism and other related fungi . The reconstructed network may serve as a valuable database for annotation of genes identified in future genome sequencing projects on aspergilli . Based on the metabolic reconstruction, a stoichiometric model was set up that includes 284 metabolites and 335 reactions, of which 268 represent biochemical conversions and 67 represent transport processes between the different intracellular compartments and between the cell and the extracellular medium . The stoichiometry of the metabolic reactions was used in combination with biosynthetic requirements for growth and pseudo-steady state mass balances over intracellular metabolites for the quantification of metabolic fluxes using metabolite balancing . This framework was employed to perform an in silico characterisation of the phenotypic behaviour of A . niger grown on different carbon sources . The effects on growth of single reaction deletions were assessed and essential biochemical reactions were identified for different carbon sources . Furthermore, application of the stoichiometric model for assessing the metabolic capabilities of A . niger to produce metabolites was evaluated by using succinate production as a case study.

Eur J Biochem, 2003 Nov, 270(22), 4555 - 63
Construction of hybrid peptide synthetases for the production of alpha-l-aspartyl-l-phenylalanine, a precursor for the high-intensity sweetener aspartame; Duerfahrt T et al.; Microorganisms produce a large number of pharmacologically and biotechnologically important peptides by using nonribosomal peptide synthetases (NRPSs) . Due to their modular arrangement and their domain organization NRPSs are particularly suitable for engineering recombinant proteins for the production of novel peptides with interesting properties . In order to compare different strategies of domain assembling and module fusions we focused on the selective construction of a set of peptide synthetases that catalyze the formation of the dipeptide alpha-l-aspartyl-l-phenylalanine (Asp-Phe), the precursor of the high-intensity sweetener alpha-l-aspartyl-l-phenylalanine methyl ester (aspartame) . The de novo design of six different Asp-Phe synthetases was achieved by fusion of Asp and Phe activating modules comprising adenylation, peptidyl carrier protein and condensation domains . Product release was ensured by a C-terminally fused thioesterase domains and quantified by HPLC/MS analysis . Significant differences of enzyme activity caused by the fusion strategies were observed . Two forms of the Asp-Phe dipeptide were detected, the expected alpha-Asp-Phe and the by-product beta-Asp-Phe . Dependent on the turnover rates ranging from 0.01-0.7 min-1, the amount of alpha-Asp-Phe was between 75 and 100% of overall product, indicating a direct correlation between the turnover numbers and the ratios of alpha-Asp-Phe to beta-Asp-Phe . Taken together these results provide useful guidelines for the rational construction of hybrid peptide synthetases.

Dev Biol (Basel), 2003, 113, 61 - 4
Current issues in validation of chromatography; Sofer G; Chromatography has now been used successfully for more than 20 years to provide the requisite purity for biologicals, but there remain some issues of concern . These issues include carryover and cleaning validation, potential for contamination by microorganisms, potential leachables, and resin lifespan . Compounding these issues is the fact that chromatography media have a very large surface area, and it is not always clear what is occurring at the surface . Furthermore, there are multiple, interactive operating parameters that have the potential to influence chromatographic performance.

Am J Vet Res, 2003 Nov, 64(11), 1421 - 8
Detection of novel chlamydiae in cats with ocular disease; von Bomhard W et al.; OBJECTIVE: To detect and characterize the full range of chlamydial infections in cats with ocular disease by use of polymerase chain reaction (PCR) assays, cytologic examination, immunohistochemical analysis, and evaluation of clinical information including status for feline herpesvirus-1 (FeHV-1) . SAMPLE POPULATION: DNA extracted from 226 conjunctival samples obtained from cats with clinically diagnosed keratitis or conjunctivitis and 30 conjunctival samples from healthy cats . PROCEDURE: PCR assays for the 16S rRNA gene specific for the order Chlamydiales and a new Chlamydophila felis (formerly Chlamydia psittaci) species-specific 23S rRNA gene were performed . Seventy-four conjunctival samples were prepared with Romanowsky-type stain, grouped on the basis of inflammatory pattern, and screened for chlamydial inclusions by use of immunohistochemical analysis . Clinical information and FeHV-1 status were recorded . RESULTS: 26 (12%) specimens had positive results for the only known feline chlamydial pathogen, C felis . Surprisingly, an additional 88 (39%) were positive for non-C felis chlamydial DNA . Identification of non-C felis chlamydial DNA by direct sequencing revealed 16S rRNA gene sequences that were 99% homologous to the sequence for Neochlamydia hartmannellae, an amebic endosymbiont . Chlamydial prevalence was significantly higher in cats with ocular disease . CONCLUSIONS AND CLINICAL RELEVANCE: Application of a broad-range detection method resulted in identification of a new agent associated with ocular disease in cats . Finding chlamydia-like agents such as N hartmannellae in coinfections with their obligate amebic host, Hartmannella vermiformis, raises questions about the potential role of these microorganisms in causation or exacerbation of ocular disease in cats.

Quintessence Int, 2003 Nov-Dec, 34(10), 733 - 8
Reestablishing biologic width with forced eruption; Felippe LA et al.; Maintenance of gingival health is one of the keys for the longevity of teeth, as well as for the longevity of restorations . In this context, the biologic width functions as a barrier against the entrance of microorganisms into the internal medium of the periodontal ligament and into the gingival and osseous connective tissue . This clinical case describes a technique to reestablish the biologic width of a central incisor using forced extrusion and done without post-treatment corrective surgery.

Biofouling, 2003 Apr, 19 Suppl, 249 - 54
Wood digestion in Pselactus spadix Herbst--a weevil attacking marine timber structures; Oevering P et al.; Pselactus spadix tunnels timber structures in the marine environment . Recent studies reported a cosmopolitan distribution for this weevil, which is frequently found in harbour and port areas . P . spadix feeds on timber (hardwood and softwood) in immature and adult life stages, but its digestion of wood components had not been investigated . Using dry weight analyses of tunnel walls and frass produced, P . spadix adults consumed Scots pine with soft rot decay at a rate of 1.59 +/- 0.37 mg d-1 and the digestibility of this substrate was 57.96 +/- 5.89 (i.e . for 100 mg consumed SR-pine, 58 mg was digested) . Using gravimetric analysis to quantify structural wood components in tunnel walls and frass, P . spadix adults were found to digest cellulose, lignin and hemicellulose with digestibility coefficients of 82.2, 41.2 and 14.5 respectively . Fourier Transform Infrared (FTIR) spectroscopy analyses of tunnel walls and frass of adults and larvae from soft rotted pine also indicated digestion of all structural components, with larvae digesting cellulose and lignin more efficiently than adults . When FTIR was employed to analyse adult tunnel walls and frass from undecayed pine, cellulose and hemicellulose were digested, but no evidence of lignin digestion was found . This study shows that adults digest lignin when soft rot is present and suggests a symbiotic function of wood degrading microorganisms.

Medicina (Kaunas), 2003, 39 Suppl 2, 55 - 9
{Octanhydroxamate of iron: synthesis, analysis and investigation of stability}; Palma P et al.; Hydroxamic acids are found as chelates in plants, as metabolites of bacteria and funges . Some aerobic microorganisms synthesize hydroxamic acids, which transmit iron from environment to the cells of the plants . Fourteen hydroxamic acids were synthesized and their interaction with iron (II and III) was investigated . The purpose of this investigation was synthesis of hydroxamic acids, creation of stable iron chelate with one of these acids, preparation of the methods of analysis, investigation of the stability of chelate and application for prophylaxis and treatment of iron deficiency anemia . We used octanhydroxamic acid for creation of chelate with iron . The synthesis of this acid was the cheapest and compound with iron appeared stable . Iron chelates after acid hydrolysis were analyzed by quantity of iron (9.8-12.3%) . We used complexometric titration method with dinatrium salt of ethylendiamintetraacetic acid without indication for quantitative analysis . Iron (II) chelate is less stable, therefore, we used Fe (III) octanhydroxamate for further investigations . Preliminary trials with rabbits confirm rapid assimilation of iron (III) chelate from gastrointestinal tract and activation of erythropoiesis.

Mol Microbiol, 2003 Nov, 50(3), 857 - 70
Cell cycle- and age-dependent activation of Sod1p drives the formation of stress resistant cell subpopulations within clonal yeast cultures; Sumner ER et al.; Phenotypic heterogeneity describes non-genetic variation that exists between individual cells within isogenic populations . The basis for such heterogeneity is not well understood, but it is evident in a wide range of cellular functions and phenotypes and may be fundamental to the fitness of microorganisms . Here we use a suite of novel assays applied to yeast, to provide an explanation for the classic example of heterogeneous resistance to stress (copper) . Cell cycle stage and replicative cell age, but not mitochondrial content, were found to be principal parameters underpinning differential Cu resistance: cell cycle-synchronized cells had relatively uniform Cu resistances, and replicative cell-age profiles differed markedly in sorted Cu-resistant and Cu-sensitive subpopulations . From a range of potential Cu-sensitive mutants, cup1Delta cells lacking Cu-metallothionein, and particularly sod1Delta cells lacking Cu, Zn-superoxide dismutase, exhibited diminished heterogeneity . Furthermore, age-dependent Cu resistance was largely abolished in cup1Delta and sod1Delta cells, whereas cell cycle-dependent Cu resistance was suppressed in sod1Delta cells . Sod1p activity oscillated approximately fivefold during the cell cycle, with peak activity coinciding with peak Cu-resistance . Thus, phenotypic heterogeneity in copper resistance is not stochastic but is driven by the progression of individual cells through the cell cycle and ageing, and is primarily dependent on only Sod1p, out of several gene products that can influence the averaged phenotype . We propose that such heterogeneity provides an important insurance mechanism for organisms; creating subpopulations that are pre-equipped for varied activities as needs may arise (e.g . when faced with stress), but without the permanent metabolic costs of constitutive expression.

Clin Microbiol Infect, 2003 Aug, 9(8), 766 - 79
The multiple roles of Fas ligand in the pathogenesis of infectious diseases; Dockrell DH; Fas ligand (FasL) is a type II transmembrane protein that plays a critical role in immune homeostasis by binding to its receptor Fas (CD95) and inducing apoptosis . Fas/FasL dysregulation contributes to infectious disease pathogenesis . Microorganisms may inhibit Fas signal transduction to prolong intracellular survival and prevent killing by immune effector cells . FasL may be upregulated in directly infected cells to enhance killing of responding immune cells and facilitate immune evasion . The host response to infection may aim to induce apoptosis in directly infected cells, but immune cells that target directly infected cells can induce Fas-mediated apoptosis of uninfected bystander cells . FasL also contributes to the generation and regulation of the inflammatory response in infection . The multiple roles of FasL in infectious disease pathogenesis are discussed in the context of viral, bacterial and parasitic infections.

Biochemistry (Mosc), 2003 Oct, 68(10), 1132 - 5
Halogenating enzymes in the biosynthesis of antibiotics; Burd VN et al.; Using blot hybridization, it has been shown that microorganisms producing halogen-containing antibiotics--Pseudomonas pyrrocinia, P . aureofaciens ACN, P . aureofaciens Pa1, P . fluorescens CHA0, Actinoplanes sp., Kitasatasporia sp., Sacharothrix aerocolonigenes, Actinomadura melliaura, and Streptomyces albogriseolus--contain the genes of the halogenating enzymes related to tryptophan-7-halogenase and monodechloroaminopyrrolnitrin halogenase from P . fluorescens BL 915.

Anal Chem, 2003 Nov 15, 75(22), 6056 - 62
Compound-specific stable isotope analysis of soil mesofauna using thermally assisted hydrolysis and methylation for ecological investigations; Evans CJ et al.; Stable isotope mass spectrometric approaches are proving to be valuable tools in unravelling biotic interactions in complex ecosystems, yielding information on trophic preferences and functional roles of individual species . Gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) provides considerable opportunities to assist in studies concerned with ecosystem processes mediated by soil invertebrates and microorganisms by determination of delta(13)C values of individual compounds, for example, lipids, amino acids etc . However, techniques conventionally adopted for "wet" chemical extractions and derivatizations necessary for compound-specific stable isotope determinations restrict the size of soil organism that can be studied and can limit investigations of individuals or even parts of individuals . We demonstrate here that individual soil mesofauna can be probed directly for their fatty acid stable isotope signatures by pyrolysis-GC/C/IRMS . A thermally assisted hydrolysis and methylation (THM) reaction is described for the determination of delta(13)C fatty acid values using trimethylsulfonium hydroxide (TMSH) . Authentic fatty acids, acyl lipids, and individual Collembola (Folsomia candida) raised on C(3) and C(4) isotopically labeled yeast were analyzed initially by py-GC/MS with TMSH and then by py-GC/C/IRMS . A kinetic isotope effect (KIE) observed with the THM reaction prevents direct calculation of the fatty acid delta(13)C values by simple mass balance equations . However, the KIE is shown to be both reproducible and robust and can therefore be accounted for by the use of correction factors . The fatty acid methyl ester compositions of individual F . candida and their respective delta(13)C values were determined and shown to agree with those obtained by conventional "wet" chemical procedures applied to much larger numbers of Collembola, thus enhancing the scope to which stable isotopes can be applied to the study of invertebrates in complex food webs in any environment.

Turk J Gastroenterol, 2003 Jun, 14(2), 145 - 7
Spontaneous bacterial peritonitis due to Brucella infection; Gursoy S et al.; Brucella infection is a systemic disease, but the microorganism rarely causes infections in the gastrointestinal system such as hepatitis, cholecystitis, colitis and pancreatitis . Spontaneous bacterial peritonitis due to Brucella is extremely rare . Herein, we report a case of cirrhosis complicated with nongranulomatous hepatitis and peritonitis, both due to Brucella . A 63 year-old man with diabetes mellitus was admitted to hospital with complaints of weakness, backache, abdominal pain and abdominal swelling . On the basis of physical examination and laboratory findings, cryptogenic cirrhosis and spontaneous bacterial peritonitis were diagnosed . Due to persistent fever and backache, serum Brucella agglutination test was performed and found to be positive . Brucella melitensis was isolated from ascitic fluid culture . Liver biopsy findings revealed cirrhosis and a nongranulomatous hepatitis which was thought might be due to Brucella infection . Doxycycline and rifampicin, in addition to diuretics were administered for spontaneous ascites infection due to Brucella . A week later, the patient's condition improved and he became afebrile . After two months of therapy, the ascites had almost disappeared.

J Vet Sci, 2000 Dec, 1(2), 105 - 11
Using pig biliary system, in vivo propagation of Enterocytozoon bieneusi, an AIDS-related zoonotic pathogen; Lee JH; A microsporidian parasite Enterocytozoon bieneusi is the most common microorganism recognized in AIDS patients, and slow scientific progress is attributed to our inability to propagate the parasite . We report upon the development of a system of propagation using the pig biliary system . The parasite spores were continuously detected in the bile samples post onset of spore shedding in the gall bladder, which suggests that this organism maintain persistent infection in the biliary system and that the hepatobiliary tree may represent a reservoir of infection . In conclusion the biliary tree is an adequate niche for the propagation of E . bieneusi . This work has also resulted in the development of a procedure of ultrasound-guided cholecystocentesis for aspirating biles . This is a simple and non-surgical procedure, and creates no signs of clinical complications in the livers and the gall bladders after dozens of separate attempts . Thus, this is a very useful and safe technique for the aspiration of bile from live animals.

Microbes Infect, 2003 Nov, 5(14), 1337 - 44
Regulation of the neutrophil-mediated inflammatory response to infection; Kobayashi SD et al.; Human polymorphonuclear leukocytes (PMNs) are the first line of defense against invading microorganisms and contribute significantly to inflammation . Recent evidence suggests that resolution of neutrophil-mediated inflammation is facilitated by an apoptosis differentiation program, a final stage of transcriptionally regulated PMN maturation that is accelerated significantly by phagocytosis.

Bioorg Chem, 2003 Dec, 31(6), 494 - 502
Reconstitution of UDP-galactopyranose mutase with 1-deaza-FAD and 5-deaza-FAD: analysis and mechanistic implications; Huang Z et al.; The galactofuranose moiety found in many surface constituents of microorganisms is derived from UDP-D-galactopyranose (UDP-Galp) via a unique ring contraction reaction catalyzed by a FAD-dependent UDP-Galp mutase . When the enzyme is reduced by sodium dithionite, its catalytic efficiency increases significantly . Since the overall transformation exhibits no net change in the redox state of the parties involved, how the enzyme-bound FAD plays an active role in the reaction mechanism is puzzling . In this paper, we report our study of the catalytic properties of UDP-Galp mutase reconstituted with deaza-FADs . It was found that the mutase reconstituted with FAD or 1-deazaFAD has comparable activity, while that reconstituted with 5-deazaFAD is catalytically inactive . Because 5-deazaFAD is restricted to net two-electron process, yet FAD and 1-deazaFAD can undergo concerted two-electron as well as stepwise one-electron redox reactions, the above results support a radical mechanism for the mutase catalyzed reaction . In addition, the activity of the mutase reconstituted with FAD was found to increase considerably at high pHs . These observations have allowed us to propose a new mechanism involving one-electron transfer from the reduced FAD to an oxocarbenium intermediate generated by C-1 elimination of UDP to give a hexose radical and a flavin semiquinone . Subsequent radical recombination leads to a coenzyme-substrate adduct which may play a central role to facilitate the opening and recyclization of the galactose ring . A deprotonation step, accompanied or followed the electron transfer step, to increase the nucleophilicity of the flavin radical anion may account for the activity enhancement at pH > 8.

J Leukoc Biol, 2004 Jan, 75(1), 3 - 4 Epub 2003 Nov 11.
Series introduction: Innate host defense of the respiratory epithelium; Hiemstra PS et al.; This series overviews the current knowledge of the innate-immune system of the lung . Special emphasis is put on mechanisms used by epithelial cells of the respiratory tract to contribute to host defense . This defense function is important, as the lung is especially prone to microbial exposure . The exchange of oxygen and carbon dioxide requires the ventilation of a large volume of environmental air that may contain respiratory pathogens . Furthermore, the digestive pathway shares with the airways the pharynx, opening another possibility for the uptake of microorganisms into the respiratory tract . Host defense is provided by a multilayered and complex system that aims to avoid colonization and infection of the lung . A classic view separates the inborn, innate-immune system from adaptive mechanisms that involve antigen-specific lymphocytes.

Arch Dis Child, 2003 Nov, 88(11), 981 - 4
The potential of recombinant surfactant protein D therapy to reduce inflammation in neonatal chronic lung disease, cystic fibrosis, and emphysema; Clark H et al.; By lowering surface tension at the air-water interface in the surfactant deficient premature lung, exogenous surfactant replacement therapy for neonatal respiratory distress syndrome has been highly successful in decreasing mortality after preterm birth . It has emerged in recent years that surfactant components not present in current surfactant formulations--particularly surfactant associated proteins A and D (SP-A and SP-D)-have additional roles in host defence distinct from the surface tension lowering effects of surfactant . SP-A and SP-D are calcium dependent carbohydrate binding proteins of the innate immune system important in the first line defence of the lung against microorganisms and in the control of lung inflammation . This review addresses the possibility that recently developed recombinant forms of SP-D could be useful therapeutically in attenuating inflammatory processes in neonatal chronic lung disease, cystic fibrosis, and emphysema.

J Ind Microbiol Biotechnol, 2003 Nov, 30(11), 627 - 35 Epub 2003 Nov 11.
Detection and characterization of chitinases and other chitin-modifying enzymes; Howard MB et al.; Multiple industrial and medical uses of chitin and its derivatives have been developed in recent years . The demand for enzymes with new or desirable properties continues to grow as additional uses of chitin, chitooligosaccharides, and chitosan become apparent . Microorganisms, the primary degraders of chitin in the environment, are a rich source of valuable chitin-modifying enzymes . This review summarizes many methods that can be used to isolate and characterize chitin-modifying enzymes including chitin depolymerases, chitodextrinases, chitin deacetylases, N-acetylglucosaminidases, chitin-binding proteins, and chitosanases . Chitin analogs, zymography, detection of reducing sugars, genomic library screening, chitooligosaccharide electrophoresis, degenerate PCR primer design, thin layer chromatography, and chitin-binding assays are discussed.

J Virol, 2003 Dec, 77(23), 12639 - 45
Blocking immune evasion as a novel approach for prevention and treatment of herpes simplex virus infection; Judson KA et al.; Many microorganisms encode immune evasion molecules to escape host defenses . Herpes simplex virus type 1 glycoprotein gC is an immunoevasin that inhibits complement activation by binding complement C3b . gC is expressed on the virus envelope and infected cell surface, which makes gC potentially accessible to blocking antibodies . Mice passively immunized with gC monoclonal antibodies prior to infection were protected against herpes simplex virus challenge only if the gC antibodies blocked C3b binding . Mice treated 1 or 2 days postinfection with gC monoclonal antibodies that block C3b binding had less severe disease than control mice treated with nonimmune immunoglobulin G (IgG) . Mice immunized with gC protein produced antibodies that blocked C3b binding to gC . Immunized mice were significantly protected against challenge by wild-type virus, but not against a gC mutant virus lacking the C3b binding domain, suggesting that protection was mediated by antibodies that target the gC immune evasion domain . IgG and complement from subjects immunized with an experimental herpes simplex virus glycoprotein gD vaccine neutralized far more mutant virus defective in immune evasion than wild-type virus, supporting the importance of immune evasion molecules in reducing vaccine potency . These results suggest that it is possible to block immune evasion domains on herpes simplex virus and that this approach has therapeutic potential and may enhance vaccine efficacy.

Environ Monit Assess, 2003 Nov, 89(1), 35 - 41
Effect of common pesticides used in the Niger Delta basin of southern Nigeria on soil microbial populations; Ekundayo EO; The effects of eleven pesticides on the populations of bacteria, actinomycetes, fungi and protozoa was investigated by treating a garden soil with their recommended rates . The microbial populations were estimated using the standard plate-count technique . Of the 11 pesticides investigated, phenylmercuric acetate (agrosan) at 50 microg g(-1) inhibited bacterial density the most, i.e . from 4,600,000 to 220 cells g(-1) . The pesticides were Pentachloronitrobenzene (PCNB), tetramethylmethylthiuram disulphide (thiram), 1-naphthylmethylcarbamate (Vetox 85), 1,2,3,4,5,6-hexachlorocyclohexane (Gammalin 20), phenylmercuric acetate (Agrosan), tetrachloroterephthalic acid (Dacthal), 4-nitrophenyl-2-nitro-4-trifluoromethylphenyl ether (Preforan), 2-ethyl-6-methyl-N-2-methoxy-1-methyl ethyl-chloroacetanide (Dual), Benlate, Brestan and Gramoxone . Pentachloronitrobenzene (PCNB) at 240,000 microg g(-1) reduced bacterial population from 4,600,000 to 2,100 cells g(-1), whereas tetramethylthiuram disulphide (thiram) at 100 microg g(-1) suppressed it by 2 log orders of magnitude . Soil application of 1-naphthylmethylcarbamate (Vetox 85) at 100 microg g(-1) and 1,2,3,4,5,6,-hexachlorocyclohexane (Gamalin 20) at 1,300 microg g(-1) repressed the bacterial numbers by 2 log orders of magnitude each . Pentachloronitrobenzene reduced the actinomycetes density from 340,000 to 320 cells g(-1) and completely eliminated all fungal and protozoan propagules from the soil . The Gammalin 20 completely wiped out all the fungi, whereas phenylmercuric acetate totally eliminated all the protozoa and reduced the fungal population from 34,000 to 60 cells g(-1) . In general, protozoa and fungi were more susceptible to fungicides than bacteria and actinomycetes . Pentachloronitrobenzene, 1,2,3,4,5,6,-hexachlorocyclohexane and phenylmercuric acetate were toxic particularly to soil microorganisms, whereas the herbicides dacthal, Preforan and Dual were quite harmless in soil at application rates of 0.1, 0.06 and 0.02 microg g(-1) respectively.

Arch Pharm Res, 2003 Oct, 26(10), 868 - 73
Differential effect of bovine serum albumin on ginsenoside metabolite-induced inhibition of alpha3beta4 nicotinic acetylcholine receptor expressed in Xenopus oocytes; Lee JH et al.; Ginsenosides, major active ingredients of Panax ginseng, that exhibit various pharmacological and physiological actions are transformed into compound K (CK) or M4 by intestinal microorganisms . CK is a metabolite derived from protopanaxadiol (PD) ginsenosides, whereas M4 is a metabolite derived from protopanaxatriol (PT) ginsenosides . Recent reports shows that ginsenosides might play a role as pro-drugs for these metabolites . In present study, we investigated the effect of bovine serum albumin (BSA), which is one of major binding proteins on various neurotransmitters, hormones, and other pharmacological agents, on ginsenoside Rg2-, CK-, or M4-induced regulation of alpha3beta4 nicotinic acetylcholine (ACh) receptor channel activity expressed in Xenopus oocytes . In the absence of BSA, treatment of ACh elicited inward peak current (I(ACh)) in oocytes expressing alpha3beta4 nicotinic ACh receptor . Co-treatment of ginsenoside Rg2, CK, or M4 with ACh inhibited I(ACh) in oocytes expressing (alpha3beta4 nicotinic ACh receptor with reversible and dose-dependent manner . In the presence of 1% BSA, treatment of ACh still elicited I(ACh) in oocytes expressing alpha3beta4 nicotinic ACh receptor and co-treatment of ginsenoside Rg2 or M4 but not CK with ACh inhibited I(ACh) in oocytes expressing alpha3beta4 nicotinic ACh receptor with reversible and dose-dependent manner . These results show that BSA interferes the action of CK rather than M4 on the inhibitory effect of I(ACh) in oocytes expressing alpha3beta4 nicotinic ACh receptor and further suggest that BSA exhibits a differential interaction on ginsenoside metabolites.

C R Biol, 2003 Aug, 326(8), 739 - 50
{Viability of heterotrophic bacteria in the Bay of Marseilles}; Gregori G et al.; Marine microorganism activities are commonly assessed by bulk methods and assigned to the total cell count . The presence in significant amounts of ghost, dead, and damaged cells makes such as assignment a non-correct one . A Nucleic Acid Double Staining protocol (NADS) of fresh water bacteria (Barbesti et al., Cytometry 40 (2000) 214-218) has been adapted to resolve viable, damaged and dead cells in marine environments (Gregori et al., Appl . Environ . Microbiol . 67 (2001) 4662-4670) . The present reports the first in situ application of this approach, conducted in the Bay of Marseilles in winter and spring periods at two sites with contrasted features.

Trends Microbiol, 2003 Nov, 11(11), 536 - 41
Genes and proteins involved in bacterial magnetic particle formation; Matsunaga T et al.; Magnetic bacteria synthesize intracellular magnetosomes that impart a cellular swimming behaviour referred to as magnetotaxis . The magnetic structures aligned in chains are postulated to function as biological compass needles allowing the bacterium to migrate along redox gradients through the Earth's geomagnetic field lines . Despite the discovery of this unique group of microorganisms 28 years ago, the mechanisms of magnetic crystal biomineralization have yet to be fully elucidated . This review describes the current knowledge of the genes and proteins involved in magnetite formation in magnetic bacteria and the biotechnological applications of biomagnetites in the interdisciplinary fields of nanobiotechnology, medicine and environmental management.

J Clin Microbiol, 2003 Nov, 41(11), 5238 - 9
Evaluation of acridine orange staining as a replacement of subcultures for BacT/ALERT-positive, gram stain-negative blood cultures; Adler H et al.; Among 18,424 blood culture sets processed during a study period of 18 months, 85 bottles that were positive by the BacT/ALERT system were Gram stain negative . Both acridine orange staining and subcultures detected microorganisms in a total of 12 bottles . Acridine orange staining can replace subcultures of false-positive blood cultures.

J Clin Microbiol, 2003 Nov, 41(11), 5153 - 8
Microarrays for genotyping human group a rotavirus by multiplex capture and type-specific primer extension; Lovmar L et al.; Human group A rotavirus (HRV) is the major cause of severe gastroenteritis in infants worldwide . HRV shares the feature of a high degree of genetic diversity with many other RNA viruses, and therefore, genotyping of this organism is more complicated than genotyping of more stable DNA viruses . We describe a novel microarray-based method that allows high-throughput genotyping of RNA viruses with a high degree of polymorphism by multiplex capture and type-specific extension on microarrays . Denatured reverse transcription (RT)-PCR products derived from two outer capsid genes of clinical isolates of HRV were hybridized to immobilized capture oligonucleotides representing the most commonly occurring P and G genotypes on a microarray . Specific primer extension of the type-specific capture oligonucleotides was applied to incorporate the fluorescent nucleotide analogue cyanine 5-labeled dUTP as a detectable label . Laser scanning and fluorescence detection of the microarrays was followed by visual or computer-assisted interpretation of the fluorescence patterns generated on the microarrays . Initially, the method detected HRV in all 40 samples and correctly determined both the G and the P genotypes of 35 of the 40 strains analyzed . After modification by inclusion of additional capture oligonucleotides specific for the initially unassigned genotypes, all genotypes could be correctly defined . The results of genotyping with the microarray fully agreed with the results obtained by nucleotide sequence analysis and sequence-specific multiplex RT-PCR . Owing to its robustness, simplicity, and general utility, the microarray-based method may gain wide applicability for the genotyping of microorganisms, including highly variable RNA and DNA viruses.

Nucleic Acids Res . 2003 Nov 15;31(22):e139.
Direct measurement of lymphocyte receptor diversity; Ogle BM et al.; The ability to mount an immune defense against infectious microorganisms and their products, and against tumors is believed to be a direct function of lymphocyte diversity . Because the diversity of lymphocyte receptor genes is >1000-fold more diverse than the entire genome and varies between genetically identical individuals, measuring lymphocyte diversity has been a daunting challenge . We developed a novel technique for measuring lymphocyte diversity directly using gene chips . We reasoned and here demonstrate that the frequency of hybridization of nucleic acids coding for lymphocyte receptors to the oligonucleotides on a gene chip varies in direct proportion to diversity . We applied the technique to detect changes in lymphocyte diversity in mice with known B cell alterations and in persons with known T cell repertoire defects . This approach is the first to provide direct analysis of lymphocyte receptor diversity and should facilitate fundamental study of the adaptive immune system and clinical efforts to assess immunological diseases . In addition, this approach could be more broadly applied, for example to measure diversity of viral quasi-species.

Mol Cell Probes, 2003 Dec, 17(6), 289 - 94
A specific PCR for the detection of Mycoplasma putrefaciens, one of the agents of the contagious agalactia syndrome of goats; Peyraud A et al.; Mycoplasma putrefaciens is listed as one of the etiologic agents of the contagious agalactia syndrome by the world organisation for animal health . This species has been characterized only recently, 1974, and the number of outbreaks caused by this microorganism so far is very scarce . It induces mastitis in infected goats although other symptoms such as arthritis in adults and septicaemia in kids are also frequently described . Up to now, the identification of M . putrefaciens relied on classical isolation and identification techniques which present a number of limitations . Specific primers for PCR have been designed based on sequence comparisons of the ArcB gene among the 'Mycoplasma mycoides cluster' and related species such as Mycoplasma cottewii and Mycoplasma yeatsii . Sequence alignments confirmed the taxonomic position of M . putrefaciens, which is related to the 'M . mycoides cluster' but also very close to M . yeatsii . The polymorphism observed amongst the different ArcB sequences allowed the determination of a primer pair yielding a specific amplification of a 316 bp-long DNA fragment by PCR . This PCR was validated in two different laboratories with a variety of mycoplasma strains isolated from goats . This new PCR technique will be very useful for a quicker determination of M . putrefaciens strains as well as a better understanding of the prevalence of M . putrefaciens infections.

Biometrics, 2003 Sep, 59(3), 505 - 11
Estimating the accuracy of polymerase chain reaction-based tests using endpoint dilution; Hughes JP et al.; Polymerase chain reaction (PCR)-based tests for various microorganisms or target DNA sequences are generally acknowledged to be highly "sensitive," yet the concept of sensitivity is ill-defined in the literature on these tests . We propose that sensitivity should be expressed as a function of the number of target DNA molecules in the sample (or specificity, when the target number is 0) . However, estimating this "sensitivity curve" is problematic, since it is difficult to construct samples with a fixed number of targets . Nonetheless, using serially diluted replicate aliquots of a known concentration of the target DNA sequence, we show that it is possible to disentangle random variations in the number of target DNA molecules from the underlying test sensitivity . We develop parametric, nonparametric, and semiparametric (spline-based) models for the sensitivity curve . The methods are compared on a new test for M . genitalium.

Environ Technol, 2003 Sep, 24(9), 1097 - 102
Toxicity of heavy metals for microorganisms isolated from slow sand filter schmutzdecke; Muhammad N et al.; This paper describes the susceptibility of three species of environmental bacteria isolated from the schmutzdecke of biologically active slow sand filters to cadmium, chromium and lead . The microorganisms, all identified as members of the genus Pseudomonas, were grown in tryptic soy broth with various concentrations of the selected heavy metals . The mean generation times (MGT) of the bacteria were compared to evaluate the toxic effects of the heavy metals . All three species tolerated high doses of heavy metals and the MGTs increased exponentially as the heavy metal concentrations increased; 12 mg l(-1) was the highest dose tested and the bacteria continued to grow albeit at very slow rates . In dilute media, the toxic effects of heavy metals were enhanced, illustrating the protection effect of organic matter and heavy metal complex formation . Growth studies of the isolated microorganisms on half-strength tryptic soy agar containing 6 mg l(-1) heavy metals also proved useful in determining toxicity.

Org Biomol Chem, 2003 Oct 21, 1(20), 3500 - 6
The first fluorogenic assay for detecting a Baeyer-Villigerase activity in microbial cells; Gutierrez MC et al.; The first fluorogenic assay allowing for detection of microbial enzymes able to perform Baeyer-Villiger oxidation is described . This is based on the use of 4-oxopentyl umbelliferyl ether 1 as a fluorogenic substrate . When Baeyer-Villigerases active against this test ketone are present in the selected whole cells, 1 is transformed into 3-hydroxypropyl umbelliferyl ether 3, which, in a subsequent step, releases the fluorescent product umbelliferone . Different microorganisms, known to be endowed with Baeyer-Villigerase activity, were assayed.

Vestn Ross Akad Med Nauk, 2003, (9), 37 - 9
{Polymerase chain reaction in the diagnosis of viral infections in the newborn}; Asadi Mobarkhan SM et al.; Two hundred and ten newborns from among a high risk group were examined for intrauterine infections by polymerase chain reaction (PCR) with the aim of assessing the clinical information density of the method in the diagnostic sphere . The patients were examined for the presence of DNA/RNA viruses and bacteria . Genetic samples of various causative agents were detected in 49% of the newborns . No correlation was established between the clinical pattern and the presence of DNA of certain microorganisms . The most severe condition of newborns was noted in cases when a DNA causative agent was found in several different biological samples . The highest mortality (in per cent) was registered in the group of newborns with detected DNA of Herpesviridae viruses . The PCR clinical value was affirmed for establishing a disease etiology in newborns.

J Biomed Mater Res B Appl Biomater, 2003 Nov 15, 67(2), 675 - 9
Electrospun nanofibrous polyurethane membrane as wound dressing; Khil MS et al.; Produced via electrospinning, polyurethane membrane, which has a unique property, has been of interest in medical fields . Electrospinning is a process by which nanofibers can be produced by an electrostatically driven jet of polymer solution . Electrospun fibers are collected in the form of membranes . The porous structured electrospun membrane is particularly important for its favorable properties: it exudates fluid from the wound, does not build up under the covering, and does not cause wound desiccation . The electrospun nanofibrous membrane shows controlled evaporative water loss, excellent oxygen permeability, and promoted fluid drainage ability, but still it can inhibit exogenous microorganism invasion because its pores are ultra-fine . Histological examination indicates that the rate of epithelialization is increased and the dermis becomes well organized if wounds are covered with electrospun nanofibrous membrane . This electrospun membrane has potential applications for wound dressing based upon its unique properties .

J Leukoc Biol, 2004 Mar, 75(3), 428 - 33 Epub 2003 Nov 03.
Shielding the double-edged sword: negative regulation of the innate immune system; Kobayashi KS et al.; The innate immune system is evolutionarily conserved among all multicellular organisms and is the first line of defense against microorganisms . It enables the host not only to combat pathogenic organisms but also to cohabit with nonpathogenic microorganisms by balancing the host-microorganism interaction . The innate immune response is activated rapidly (within hours) compared with adaptive immunity . Activation of the innate immune system allows the activation of the adaptive immune response by production of proinflammatory cytokines and by providing stimulatory signals via major histocompatibility complex molecules and costimulatory molecules such as CD40, CD80, or CD86; together, these lead to the full activation of both immune systems to fight against pathogenic microorganisms . Activation of the innate immune system, however, can be a double-edged sword for the host . Proinflammatory cytokines mediate a positive feedback loop on the innate immune system, and overproduction of cytokines, if unchecked, is hazardous to the host and may cause severe outcomes such as hyperthermia, organ failure, and even death in extreme cases . Moreover, if the overproduction of proinflammatory cytokines persists, it may cause chronic inflammatory diseases . During evolution, the innate immune system has acquired complicated regulatory systems to control itself so that this "sword" will not kill the host . Various mechanisms including inhibition of Toll-like receptor signaling by interleukin-1 receptor-associated kinase-M have evolved for this purpose and are important not only to fight against pathogenic microorganisms efficiently but also are critical for the peaceful coexistence with commensal bacterial flora.

Riv Biol, 2003 May-Aug, 96(2), 317 - 26
The microbial DNA cycle in soil; England LS et al.; Upon microbial cell death and lysis in soil, the free or naked DNA is exposed to the dynamic environment of the soil . The DNA can be enzymatically degraded by nucleases (DNases), bind to soil components, genetically transform competent bacterial cells and be a nutrient for other microorganisms . In this article we discuss the dual role of DNA as genetic material and as a nutrient source in the soil environment.

Riv Biol, 2003 May-Aug, 96(2), 271 - 91
Measure of species variability for a microbial taxonomy based on the relative resemblance; Cardinali G; The concept of species currently in use in microbial taxonomy is based on sex because derives from that developed in zoology and botany . The absence of sex as the only system to reproduce does not allow to use the hybridization as the test to assess the conspecificity of microbial strains, forcing microbial taxonomists to use relative resemblance among strains as the only tool to define microbial species and to classify new microorganisms . Relative resemblance can be intuitively defined as the situation in which two strains of the same species must be more similar than each of them with a strain of any other species . Unfortunately, there are several algorithms to define the similarity between two strains, but none can be used to estimate the average distance between several members of the same species . This paper describes the problems inherent with the definition of species without hybridization tests and proposes two algorithms for a standard estimation of the overall variability of a species with the data obtained from a sample of strains . These measures will allow the non-subjective determination of the overall similarity among members of the same species using results from both phenotypic and molecular analyses . Both algorithms are based on a bootsrapping procedure implemented by RHO and SMA, two software applications freely available from the internet . These applications allow an easy evaluation of parameters such as the overall variability and levels of confidence associated with the sample of strains and the panel of characters under study.

Riv Biol, 2003 May-Aug, 96(2), 241 - 60
The contribution of plant biology to the concept of virus (1886-1917); Pennazio S; Between 1860 and 1880 the so-called "theory of the infective germ", which stated in final way that every infectious disease was produced by a living pathogen agent, achieved great consent . The criteria of determining the presence of infectious pathogens (fungi, bacteria, protozoa) were established by "Koch's postulate", a set of experimental procedures conceived for isolating and determining single pathogens . In the last quarter of the 19th century became however evident that the agents of severe infectious diseases could not be identified through the "postulates" . These agents could not be seen in light microscopy nor cultured in vitro but could pass through the thin pores of filters which hold back cellular micro-organisms . This last characteristic became a selective method to recognise these peculiar agents, from then named "filterable viruses" . Most microbiologists considered the filterable viruses as living micro-organisms because of their extraordinary capacity of in vivo proliferation, and the impossibility of pointing out their structures was due to limits of the experimental techniques . Between the end of the 19th century and 1917, four plant biologists suggested that the filterable viruses were complex chemical substances rather than cellular microorganisms . Their contribution, not appreciated by the contemporary colleagues, laid the foundation of the modern concept of virus.

Biotechnol Bioeng, 2003 Dec 20, 84(6), 700 - 9
Data reconciliation and parameter estimation in flux-balance analysis; Raghunathan AU et al.; Flux blance analysis (FBA) has been shown to be a very effective tool to interpret and predict the metabolism of various microorganisms when the set of available measurements is not sufficient to determine the fluxes within the cell . In this methodology, an underdetermined stoichiometric model is solved using a linear programming (LP) approach . The predictions of FBA models can be improved if noisy measurements are checked for consistency, and these in turn are used to estimate model parameters . In this work, a formal methodology for data reconciliation and parameter estimation with underdetermined stoichiometric models is developed and assessed . The procedure is formulated as a nonlinear optimization problem, where the LP is transformed into a set of nonlinear constraints . However, some of these constraints violate standard regularity conditions, making the direct numerical solution very difficult . Hence, a barrier formulation is used to represent these constraints, and an iterative procedure is defined that allows solving the problem to the desired degree of convergence . This methodology is assessed using a stoichiometric yeast model . The procedure is used for data reconciliation where more reliable estimations of noisy measurements are computed . On the other hand, assuming unknown biomass composition, the procedure is applied for simultaneous data reconciliation and biomass composition estimation . In both cases it is verified that the f measurements required to get unbiased and reliable estimations is reduced if the LP approach is included as additional constraints in the optimization .

Biotechnol Bioeng, 2003 Dec 20, 84(6), 627 - 38
Non-thermal bacterial inactivation with dense CO(2); Spilimbergo S et al.; The use of CO(2) under pressure (dense CO(2)) is one of the most promising techniques to achieve cold pasteurization and/or sterilization of liquid and solid materials, and is likely to replace or partially substitute currently and widely applied thermal processes . Although the ability of CO(2) to inactivate microorganisms has been known since the 1950's, only within the last 15 years it has received special attention, and the scientific and economic interest towards practical applications is presently growing more and more . Here we collect and discuss the relevant current knowledge about the potentials of dense CO(2) as a non-thermal technology in the field of microbial inactivation . We summarize the state of the art, including definitions, description of the equipment, relevant applications, in both simple suspensions and complex media, for the treatment of a wide range of microorganisms in both liquid and solid substrates . Finally, we also summarize and discuss the different hypotheses about the mechanisms of inactivation .

Appl Microbiol Biotechnol, 2004 Apr, 64(3), 410 - 5 Epub 2003 Nov 01.
A thermodynamic interpretation of cell hydrophobicity in aerobic granulation; Liu Y et al.; Aerobic granulation can be regarded as a microorganism-to-microorganism self-immobilization process, in which cell hydrophobicity could be a decisive parameter in determining the microorganism-to-microorganism interaction and structural compactness of aerobic granules . This study looked into the thermodynamic interpretation of cell hydrophobicity in aerobic granulation; and a model that correlates microbial interaction and relative cell hydrophobicity defined as the ratio of cell hydrophobicity over cell hydrophilicity was derived . This model describes how cell hydrophobic and hydrophilic interactions affect aerobic granulation and offers deep insights into the thermodynamic mechanisms of microbial aggregation . The model prediction was in good agreement with experimental data . Results showed that aerobic granulation was a function of cell hydrophobicity over cell hydrophilicity, i.e . a high cell hydrophobicity strongly favors microbial aggregation and results in a more compact structure.

Curr Top Microbiol Immunol, 2004, 282, 1 - 30
Phosphoinositide involvement in phagocytosis and phagosome maturation; Botelho RJ et al.; Cells of the innate immune system engulf invading microorganisms into plasma membrane-derived vacuoles called phagosomes . Newly formed phagosomes gradually acquire microbicidal properties by a maturation process which involves sequential and coordinated rounds of fusion with endomembranes and concomitant fission . Some pathogens interfere with this maturation sequence and thereby evade killing by the immune cells, managing to survive intracellularly as parasites . Phosphoinositides seem to be intimately involved in the processes of phagosome formation and maturation, and initial observations suggest that the ability of some microorganisms to survive intracellularly is associated with alterations in phosphoinositide metabolism . This chapter presents a brief overview of phosphoinositides in cells of the immune system, their metabolism in the context of phagocytosis and phagosome maturation and their possible derangements during infectious pathogenosis.

Mikrobiyol Bul, 2003 Apr-Jun, 37(2-3), 215 - 21
{Medical importance of yeast killer toxin}; Cerikcioglu N; Yeast killer toxins (YKT) are exotoxins produced by several yeasts including certain Candida species . They can kill fungi, bacteria and even protozoa by binding to the specific receptors on their cell surfaces . On the basis of their killer effect, studies have been carried out on the differentiation of fungi and some bacteria by using YKT as epidemiological tools . Following biotyping procedures, idiotypic and anti-idiotypic antibodies produced against YKT have been shown to exhibit great potential in protection and immunotherapy against several microorganisms, promising as novel approaches to fight against microbial diseases . In this review article, the structural properties, ecologic and epidemiologic importance, therapeutic, prophylactic and immunotherapy studies of yeast killer toxins, have been discussed.

Crit Rev Microbiol, 2003, 29(3), 215 - 21
Cell wall-deficient (CWD) bacterial pathogens: could amylotrophic lateral sclerosis (ALS) be due to one?
Koch AL.
Recently, a number of diseases that had been thought previously to be caused by something other than an infectious agent are now known to be caused by bacteria . It now appears that it is not uncommon that bacteria, viruses, or fungi can cause diseases even when these organisms have not been detected or cultured . The most recent, well-publicized case is that of stomach ulcers; these are largely due to Helicobacter pylori infections . Here, the possibility is explored that amylotrophic lateral sclerosis (ALS) is caused by a cell wall-deficient microorganism.

Biofizika, 2003 Sep-Oct, 48(5), 940 - 9
{Spectral studies of a single cell of marine microplankton . History and perspectives}; Karnaukhov VN et al.; Methods and instruments for the spectral analysis of single cells of sea microplankton under the conditions of expeditions to the Mediterranean Sea and tropical zone of the Atlantic Ocean were proposed . Special emphasis was given to the ways of adaptation of sea microplankton communities to unfavorable environment with the formation of autotrophic and heterotrophic symbiont organisms (Radiolarian, Foraminifera, etc.) having closed cycles of phosphate metabolism and variotrophic microorganisms (Cyanophyceae and Dinoflagellata class).

Genetika, 2003 Sep, 39(9), 1180 - 4
{Induction of SOS-response of cells exposed to autoregulatory factors of microorganisms}; Margulis AB et al.; Among examined microbial growth regulators of alkyl hydroxybenzene group (hexylresorcinol, methylresorcinol, and hydroxyethylphenol), only hexylresorcinol induces cellular SOS response, demonstrating a dose-dependent increase of the induction factor in the SOS chromotest with the Escherichia coli PQ37 strain . At the highest of used concentrations (100 micrograms/ml), hydroxyethylphenol and nonalkylated resorcinol were shown to exert a weak toxic effect, reducing the activity of constitutive alkaline phosphatase, but did not induce SOS response . Nontoxic methylresorcinol did not induce genome damage, which can trigger SOS functions . It is concluded that substitutions in phenolic ring affect genotoxic activity of alkylresorcinols.

Chemosphere, 2004 Jan, 54(4), 569 - 74
Plant uptake of aldicarb from contaminated soil and its enhanced degradation in the rhizosphere; Sun H et al.; Experiments were conducted to investigate the degradation of aldicarb, an oxime carbamate insecticide, in sterile, non-sterile and plant-grown soils, and the capability of different plant species to accumulate the pesticide . The degradation of aldicarb in soil followed first-order kinetics . Half lives (t1/2) of aldicarb in sterile and non-sterile soil were 12.0 and 2.7 days, respectively, which indicated that microorganisms played an important part in the degradation of aldicarb in soil . Aldicarb disappeared more quickly (p< or =0.05) in the soil with the presence of plants, and t1/2 of the pesticide were 1.6, 1.4 and 1.7 days in the soil grown with corn, mung bean and cowpea, respectively . Comparison of plant-promoted degradation and plant uptake showed that the enhanced removal of aldicarb in plant-grown soil was mainly due to plant-promoted degradation in the rhizosphere.

Bioessays, 2003 Nov, 25(11), 1119 - 28
Pathogenic archaea: do they exist?
Cavicchioli R, Curmi PM, Saunders N, Thomas T.
Archaea are microorganisms that are distinct from bacteria and eukaryotes . They are prevalent in extreme environments, and yet found in most ecosystems . They are a natural component of the microbiota of most, if not all, humans and other animals . Despite their ubiquity and close association with humans, animals and plants, no pathogenic archaea have been identified . Because no archaeal pathogens have yet been identified, there is a general assumption that archaeal pathogens do not exist . This review examines whether this is a good assumption by investigating the potential for archaea to be or become pathogens . This is achieved by addressing: the diversity of archaea versus known pathogens, opportunities for archaea to demonstrate pathogenicity and be detected as pathogens, reports linking archaea with disease, and immune responses to archaea . In addition, molecular and genomic data are examined for the presence of systems utilised in pathogenesis . The view of this report is that, although archaea can presently be described as non-pathogenic, they have the potential to be (discovered as) pathogens . The present optimistic view that there are no archaeal pathogens is tainted by a severe lack of relevant knowledge, which may have important consequences in the future .

Astrobiology, 2003 Summer, 3(2), 393 - 406
Temperature and moisture conditions for life in the extreme arid region of the Atacama desert: four years of observations including the El Niño of 1997-1998; McKay CP et al.; The Atacama along the Pacific Coast of Chile and Peru is one of the driest and possibly oldest deserts in the world . It represents an extreme habitat for life on Earth and is an analog for life in dry conditions on Mars . We report on four years (September 1994-October 1998) of climate and moisture data from the extreme arid region of the Atacama . Our data are focused on understanding moisture sources and their role in creating suitable environments for photosynthetic microorganisms in the desert surface . The average air temperature was 16.5 degrees C and 16.6 degrees C in 1995 and 1996, respectively . The maximum air temperature recorded was 37.9 degrees C, and the minimum was -5.7 degrees C . Annual average sunlight was 336 and 335 W m(-2) in 1995 and 1996, respectively . Winds averaged a few meters per second, with strong fohn winds coming from the west exceeding 12 m s(-1) . During our 4 years of observation there was only one significant rain event of 2.3 mm, which occurred near midnight local time . We suggest that this event was a rainout of a heavy fog . It is of interest that the strong El Nino of 1997-1998 brought heavy rainfall to the deserts of Peru, but did not bring significant rain to the central Atacama in Chile . Dew occurred at our station frequently following high nighttime relative humidity, but is not a significant source of moisture in the soil or under stones . Groundwater also does not contribute to surface moisture . Only the one rain event of 2.3 mm resulted in liquid water in the soil and beneath stones for a total of only 65-85 h over 4 years . The paucity of liquid water under stones is consistent with the apparent absence of hypolithic (under-stone) cyanobacteria, the only known primary producers in such extreme deserts.

Astrobiology, 2003 Summer, 3(2), 343 - 50
Subfreezing activity of microorganisms and the potential habitability of Mars' polar regions; Jakosky BM et al.; The availability of water-ice at the surface in the Mars polar cap and within the top meter of the high-latitude regolith raises the question of whether liquid water can exist there under some circumstances and possibly support the existence of biota . We examine the minimum temperatures at which liquid water can exist at ice grain-dust grain and ice grain-ice grain contacts, the minimum subfreezing temperatures at which terrestrial organisms can grow or multiply, and the maximum temperatures that can occur in martian high-latitude and polar regions, to see if there is overlap . Liquid water can exist at grain contacts above about -20 degrees C . Measurements of growth in organisms isolated from Siberian permafrost indicate growth at -10 degrees C and metabolism at -20 degrees C . Mars polar and high-latitude temperatures rise above -20 degrees C at obliquities greater than ~40 degrees, and under some conditions rise above 0 degrees C . Thus, the environment in the Mars polar regions has overlapped habitable conditions within relatively recent epochs, and Mars appears to be on the edge of being habitable at present . The easy accessibility of the polar surface layer relative to the deep subsurface make these viable locations to search for evidence of life.

Life Sci, 2003 Nov 21, 74(1), 87 - 97
A novel and potent biological antioxidant, Kinobeon A, from cell culture of safflower; Kanehira T et al.; Kinobeon A was originally isolated from cultured cells of safflower (Carthamus tinctorius L., Compositae) . It had never previously been directly isolated from safflower or other plants, animals or microorganisms . In this report, we demonstrate the anti-oxidative effects of kinobeon A and compare the results with those two known natural antioxidants, lignan (nordihydroguaiaretic acid) and quercetin . The NADPH-induced microsomal lipid peroxidation system was employed to assess anti-oxidative effects of kinobeon A . Addition of kinobeon A to the system significantly decreased the formation of thiobarbituric acid reactive substances (TBARS) in a dose-dependent manner with effects similar to those of lignan and quercetin . Formation of TBARS was completely inhibited at 10 microM of kinobeon A . Employing the xanthine/xanthine oxidase/nitroblue tetrazolium system and the KO2/XTT system, the superoxide anion scavenging activity of kinobeon A was greater than that of lignan or quercetin . IC50 values calculated for kinobeon A in these two systems were 1 microM and 0.8 microM, respectively . Kinobeon A exerted cytoprotective effects following oxidative treatments with hydrogen peroxide, cumene hydroperoxide, menadione and xanthine oxidase (XOD) . Addition of kinobeon A to the systems markedly enhanced survival ratios of Madin-Darby bovine kidney cells, while their survival significantly decreased with the oxidative treatment alone . Kinobeon A exhibited stronger effect on the cell viability than lignan or quercetin when menadion or XOD were used as inducing reagents of oxidative stress . The present study demonstrates for the first time that kinobeon A prevents oxidative stresses and could be a useful cytoprotective reagent.

Chemosphere, 2004 Jan, 54(3), 225 - 33
The degradability of biodegradable plastics in aerobic and anaerobic waste landfill model reactors; Ishigaki T et al.; Degradabilities of four kinds of commercial biodegradable plastics (BPs), polyhydroxybutyrate and hydroxyvalerate (PHBV) plastic, polycaprolactone plastic (PCL), blend of starch and polyvinyl alcohol (SPVA) plastic and cellulose acetate (CA) plastic were investigated in waste landfill model reactors that were operated as anaerobically and aerobically . The application of forced aeration to the landfill reactor for supplying aerobic condition could potentially stimulate polymer-degrading microorganisms . However, the individual degradation behavior of BPs under the aerobic condition was completely different . PCL, a chemically synthesized BP, showed film breakage under the both conditions, which may have contributed to a reduction in the waste volume regardless of aerobic or anaerobic conditions . Effective degradation of PHBV plastic was observed in the aerobic condition, though insufficient degradation was observed in the anaerobic condition . But the aeration did not contribute much to accelerate the volume reduction of SPVA plastic and CA plastic . It could be said that the recalcitrant portions of the plastics such as polyvinyl alcohol in SPVA plastic and the highly substituted CA in CA plastic prevented the BP from degradation . These results indicated existence of the great variations in the degradability of BPs in aerobic and anaerobic waste landfills, and suggest that suitable technologies for managing the waste landfill must be combined with utilization of BPs in order to enhance the reduction of waste volume in landfill sites.

Biotechnol Bioeng, 2003 Dec 5, 84(5), 544 - 51
Swirling flow implementation in a photobioreactor for batch and continuous cultures of Porphyridium cruentum; Muller-Feuga A et al.; Light is the main limiting factor in photoautotrophic-intensive production of microorganisms, and improvement of its use is an important concern for photobioreactor design and operation . Swirling flows, which are known to improve mass and photon transfers, were applied to annular light chambers of a photobioreactor and studied by simulation and microalgal culture . Two hydrodynamic conditions were compared: axial flow generating poor radial mixing, and tangential flow generating three-dimensional swirling motion . Batch and continuous cultures of the Rhodophyte Porphyridium cruentum were performed in a 100-L, 1.5-m(2), fully controlled photobioreactor with eight light chambers . The inlet design of these chambers was modified to create the hydrodynamic conditions for comparison . Various intensities of swirling motion were used, characterized by the velocity factor (VF), defined as the ratio between annular chamber flow and inlet aperture sections . Experiments were performed within the range of photon flux densities (PFD) optimizing the yield of light energy transformation into living substance for the species and the temperature used . Culture kinetics with swirling flows generated by apertures of VF = 2, 4, and 9 were compared with pseudoaxial VF = 2 chosen as reference . Batch cultures with VF = 4 swirling flow showed no significant difference, whereas continuous cultures proved more discriminating . Although no significant difference was obtained for VF = 2, a 7% increase of steady-state productivity and a 26% decrease in time required to reach this steady state were obtained with VF = 4 swirling flow . This beneficial effect of swirling flow could have accounted for increased mixing . Conversely, VF = 9 swirling flow resulted in a 9% decrease of steady-state productivity and a 9% increase in the time required to reach this steady state, a negative effect that could have accounted for increased shear stress . CO(2) bioconversion yield at steady state showed a 34% increase for VF = 4 . These results suggest that swirling motion makes microalgal cultures more efficient, provided that the resulting adverse effects remain acceptable . Experimental investigation was completed by a theoretical approach in which simulation of continuous cultures of P . cruentum was based on the hydrodynamic conditions achieved in the photobioreactor . Although the results obtained with pseudoaxial flow were correctly predicted, simulations with swirling flow showed a marked enhancement of productivity not observed experimentally . The influence of side effects induced by increased mixing (particularly hydrodynamic shear stress) was considered with respect to modeling assumptions . Comparison of experimental results with theoretical simulation provided a better understanding of the mixing effect, a key factor in improving the efficiency of such bioprocesses .

Curr Opin Microbiol, 2003 Oct, 6(5), 446 - 51
Multidrug efflux pumps and resistance: regulation and evolution; Paulsen IT; Multidrug transporters have long puzzled researchers because of their ability to extrude multiple structurally dissimilar toxic chemotherapeutic agents . They appear to be essentially ubiquitous with many microorganisms possessing large arsenals of predicted multidrug efflux transporters . Recent reports on the structures of multidrug efflux transporters and their cognate regulators have suggested that the basis of multidrug recognition is the presence of large flexible hydrophobic cavities capable of accommodating different compounds that are bound via hydrogen bonding and/or electrostatic interactions . This structural data provides the context for understanding other questions about the evolution of multidrug transporters and their natural physiological roles.

Enferm Infecc Microbiol Clin, 2003 Nov, 21(9), 484 - 7
{In vitro activity of voriconazole and three other antifungal agents against dermatophytes}; Serrano-Martino Mdel C et al.; INTRODUCTION: The increase in infections due to dermatophytes in recent years led us to study the effectiveness of new antifungal formulations against these microorganisms . METHODS: The in vitro activity of a new antifungal agent, voriconazole, was compared with three other antifungal agents, itraconazole, fluconazole and terbinafine, against 120 dermatophytes belonging to four species (61 Trichophyton mentagrophytes, 34 Microsporum canis, 13 M . gypseum and 12 T . rubrum) . A broth microdilution method was used following the recommendations of the NCCLS document M38-P with some modifications . RESULTS: Terbinafine was the most active agent against the dermatophytes studied (MIC90 < or = 0.03 mg/ml), followed by voriconazole (MIC90, 0.25 micro g/ml) and itraconazole (MIC90, 0.5 micro g/ml) . Fluconazole was the least active antifungal agent . The most susceptible species was M . canis . CONCLUSIONS: Voriconazole was found to have effective activity against dermatophytes.

Anal Chem, 2003 Aug 1, 75(15), 3817 - 22
Microorganism identification by matrix-assisted laser/desorption ionization mass spectrometry and model-derived ribosomal protein biomarkers; Pineda FJ et al.; An improved data analysis method is described for rapid identification of intact microorganisms from MALDI-TOF-MS data . The method makes no use of mass spectral fingerprints . Instead, a microorganism database is automatically generated that contains biomarker masses derived from ribosomal protein sequences and a model of N-terminal Met loss . We quantitatively validate the method via a blind study that seeks to identify microorganisms with known ribosomal protein sequences . We also include in the database microorganisms with incompletely known sets of ribosomal proteins to test the specificity of the method . With an optimal MALDI protocol, and at the 95% confidence level, microorganisms represented in the database with 20 or more biomarkers (i.e., those with complete or nearly completely sequenced genomes) are correctly identified from their spectra 100% of the time, with no incorrect identifications . Microorganisms with seven or less biomarkers (i.e., incompletely sequenced genomes) are either not identified or misidentified . Robustness with respect to variations in sample preparation protocol and mass analysis protocol is demonstrated by collecting data with two different matrixes and under two different ion-mode configurations . Statistical analysis suggests that, even without further improvement, the method described here would successfully scale up to microorganism databases with roughly 1000 microorganisms . The results demonstrate that microorganism identification based on proteome data and modeling can perform as well as methods based on mass spectral fingerprinting.

FEMS Microbiol Lett, 2003 Oct 10, 227(1), 87 - 92
A generalized transducing thiophage (TPC-1) of a facultative sulfur chemolithotrophic bacterium, Bosea thiooxidans CT5, of alpha-Proteobacteria, isolated from Indian soil; Deb C et al.; We have isolated and characterized a double-stranded DNA bacteriophage (TPC-1) of Bosea thiooxidans, a facultative sulfur chemolithotrophic bacterium . The name 'thiophage' is introduced for phage(s) infecting sulfur chemolithotrophic bacteria . Electron micrographs showed the phage particle with an icosahedral head and a very short wedge-like tail . TPC-1 is classified as the C1 morphotype of the Podoviridae family . Restriction map and terminal ends detection by end fill labeling of the TPC-1 genomic DNA showed that the genome is linear with 5' protruding cohesive termini . Contour length mapping of the DNA genome also revealed it to be a linear fragment with size ( approximately 44 kb) corresponding with the size estimated from restriction fragment analyses and proved the non-redundant nature of the linear genome topology . In colorless sulfur chemolithotrophic microorganisms, TPC-1 is the first report of a generalized transducing thiophage.

J Biomol Screen, 2003 Oct, 8(5), 559 - 65
A high-throughput amenable colorimetric assay for enantioselective screening of nitrilase-producing microorganisms using pH sensitive indicators; Banerjee A et al.; Based on the color change of an indicator due to the release of hydrogen ion from a nitrilase-catalyzed reaction, a rapid colorimetric method was established for the enantioselective screening of nitrilase-producing microorganisms . The formation of acids due to the nitrilase-mediated hydrolysis of nitriles causes a drop in the pH, which in turn results in a change of color of the solution (containing indicator) that can be observed visually . The buffer (0.01 M phosphate, pH 7.2) and indicator (Bromothymol blue, 0.01%) were selected in such a way that both have the same affinity for the released protons . The enantioselectivity of nitrilases was estimated by comparing the hydrolysis of (R)-mandelonitrile with that of racemate under the same conditions . The method was used to screen a library of nitrilase-producing microorganisms, isolated in the authors' laboratory for their ability to enantioselectively hydrolyze mandelonitrile to mandelic acid, an important chiral building block.

Internist (Berl), 2003 Sep, 44(9), 1180 - 5
{A 41 year-old male patient with fever of unknown origin and bacteremia with actinobacillus actinomycetemcomitans}; Rosenkranz S et al.; A 41-year-old male patient presented with fever of unknown origin which had been present for 5 months . The primary diagnostic procedures did not identify an infectious focus . After a lag phase of 13 days, blood cultures became positive for Actinobacillus actinomycetemcomitans, which belongs to the HACEK group of microorganisms . According to the DUKE criteria, infective endocarditis was diagnosed despite negative transesophageal echocardiography (TEE) . The sensitivity of TEE is estimated between 86 and 94% . An infected tooth was extracted as a possible focus, and the patient was treated with i.v . antibiotics for 5 weeks according to the guidelines of the AHA and ACC . The fever was permanently terminated, and the further course of the patient was uneventful . Despite negative TEE, the diagnosis "infective endocarditis" should not be rejected, and blood cultures should be grown for up to 30 days.

Appl Microbiol Biotechnol, 2003 Dec, 63(3), 249 - 57 Epub 2003 Oct 18.
Bioleaching review part B: progress in bioleaching: applications of microbial processes by the minerals industries; Olson GJ et al.; This review describes the historical development and current state of metals leaching and sulfide mineral biooxidation by the minerals industries . During the past 20 years commercial processes employing microorganisms for mineral recovery have progressed from rather uncontrolled copper dump leaching to mineral oxidation and leaching in designed bioheaps for oxidation of refractory gold ores and for copper recovery . Also during this period of time, stirred tank bioleaching has been commercialized for cobalt recovery and for biooxidation of refractory gold ores . Chalcopyrite bioleaching in stirred tanks is on the verge of commercialization . Commercial applications of biohydrometallurgy have advanced due to favorable process economics and, in some cases, reduced environmental problems compared to conventional metal recovery processes such as smelting . Process development has included recognition of the importance of aeration of bioheaps, and improvements in stirred tank reactor design and operation . Concurrently, knowledge of the key microorganisms involved in these processes has advanced, aided by advances in molecular biology to characterize microbial populations.

ORL J Otorhinolaryngol Relat Spec, 2003 Jul-Aug, 65(4), 202 - 5
Local pathogenic bacteria in allergic rhinitis: a novel concept of its pathogenesis; Gluck U et al.; BACKGROUND: It is still unclear to what extent microorganisms are involved in the pathogenesis of allergic rhinitis . OBJECTIVE: Therefore, we examined the mucosal colonization with potential pathogenic bacteria (PPB) of the nasal cavity in allergic and nonallergic subjects . METHODS: In an open prospective study of 389 office workers (297 men, mean age 42.5 years, and 92 women, mean age 36.7 years), bacterial swabs were taken selectively from both nasal cavities . Standard skin tests for various aeroallergens and negative control tests were conducted in parallel in these subjects . RESULTS: In the 389 subjects, we found positive skin tests in 58 (15%); 37 of these revealed a high level of sensitivity, whilst the other 21 persons had low to moderate levels . Ninety percent of the 58 sensitive persons had PPB in their nasal cavity while only 36% (119) of the remaining 331 subjects with negative skin tests were shown to have PPB in nasal cultures (p < 0.001) . Sixty-four percent of the sensitized subjects with PPB were found to have more than 2 PPB species and a positive correlation with the intensity of the skin reaction . In contrast, only 18% (22) of the 119 nonallergic test persons with PPB had more than 1 PPB species . CONCLUSIONS: This finding of an unusually high frequency of nasal PPB in subjects with positive skin tests to aeroallergens may indicate an involvement of PPB in the pathogenesis of allergic rhinitis . Yet, further data are still lacking to support this novel concept .

J Bacteriol, 2003 Nov, 185(21), 6385 - 91
Investigation of the role of electrostatic charge in activation of the Escherichia coli response regulator CheY; Smith JG et al.; In a two-component regulatory system, an important means of signal transduction in microorganisms, a sensor kinase phosphorylates a response regulator protein on an aspartyl residue, resulting in activation . The active site of the response regulator is highly charged (containing a lysine, the phosphorylatable aspartate, two additional aspartates involved in metal binding, and an Mg(2+) ion), and introduction of the dianionic phosphoryl group results in the repositioning of charged moieties . Furthermore, substitution of one of the Mg(2+)-coordinating aspartates with lysine or arginine in the Escherichia coli chemotaxis response regulator CheY results in phosphorylation-independent activation . In order to examine the consequences of altered charge distribution for response regulator activity and to identify possible additional amino acid substitutions that result in phosphorylation-independent activation, we made 61 CheY mutants in which residues close to the site of phosphorylation (Asp57) were replaced by various charged amino acids . Most substitutions (47 of 61) resulted in the complete loss of CheY activity, as measured by the inability to support clockwise flagellar rotation . However, 10 substitutions, all introducing a new positive charge, resulted in the loss of chemotaxis but in the retention of some clockwise flagellar rotation . Of the mutants in this set, only the previously identified CheY13DK and CheY13DR mutants displayed clockwise activity in the absence of the CheA sensor kinase . The absence of negatively charged substitution mutants with residual activity suggests that the introduction of additional negative charges into the active site is particularly deleterious for CheY function . Finally, the spatial distribution of positions at which amino acid substitutions are functionally tolerated or not tolerated is consistent with the presently accepted mechanism of response regulator activation and further suggests a possible role for Met17 in signal transduction by CheY.

Immunity, 2003 Oct, 19(4), 561 - 9
Natural killer cells activated by MHC class I(low) targets prime dendritic cells to induce protective CD8 T cell responses; Mocikat R et al.; Conserved molecular patterns derived from pathogenic microorganisms prime antigen-presenting dendritic cells (DC) to induce adaptive T cell responses . In contrast, virus-infected or tumor cells that express low levels of major histocompatibility complex (MHC) class I activate natural killer (NK) cells for direct killing . It is unknown whether NK cell recognition of MHC class I(low) targets can also induce adaptive T cell responses . Here, we show that MHC class I(low) targets initiate a cascade of immune responses, starting with the immediate activation of NK cells . The activated NK cells then prime DC to produce IL-12 and to induce highly protective CD8 T cell memory responses . Therefore, sensing of MHC class I(low) targets by NK cells can link innate and adaptive immunity to induce protective T cell responses and may alarm the immune system during early infection with noncytopathic viruses.

J Environ Sci (China), 2003 Sep, 15(5), 590 - 4
Analysis of parental strain DNA fragments existing in GEMs-Fhhh; Hao CB et al.; There were 6 target DNA fragments of the three parental strains existing in the cell of GEMs (genetically engineered microorganism strain) Fhhh measured in this research by PCR(polymerase chain reaction) . The determination showed that GEMs Fhhh contained all the 6 target DNA fragments, mnp1, mnp2, lip1, lip2, FLO1 and 16S rDNA, and had the molecular genetic stability . Meanwhile the PCR production of each parental strain could only had its target DNA fragments and was different from each other . It may illustrate that the technique of the inter-kingdom protoplast fusion for the construction of GEMs Fhhh through the process of intercellular gene recombination could be used as a reliable bioengineering technique to create the specific functional stain for the pollution control.

Eur J Med Chem, 2003 Sep, 38(9), 837 - 40
Synthesis of some novel imidazolinones as potent anticonvulsant agents; Joshi H et al.; Imidazolinone derivatives of IIa-IIc, IIIa-IIIf and IVa-IVf have been synthesised by the condensation of some known sulpha drugs with 5-oxazolone derivatives, which were prepared by Erlenmeyer condensation of benzoyl glycine with different aldehydes in the presence of sodium acetate and acetic anhydride . The constitution of the products has been supported by elemental analysis and IR, 1H-NMR spectral data . The products have been screened for their (a) in vitro growth inhibitory activity against several microorganisms and (b) in vivo anticonvulsant activity.

Arch Pharm Res, 2003 Sep, 26(9), 686 - 96
Synthesis and biological investigations of new thiazolidinone and oxadiazoline coumarin derivatives; Abd Elhafez OM et al.; Ethyl (coumarin-4-oxy)acetate 1 was prepared through the reaction of 4-hydroxycoumarin with ethyl bromoacetate . Compound 1 was allowed to react with hydrazine hydrate to produce coumarin-4-oxyacetic hydrazide 2 . The synthesis of N-(arylidene and alkylidene)-coumarin-4-oxyacetic hydrazones 3-20 was performed . The preparation of 2-substituted-3-{(coumarin-4-oxy) acetamido}thiazolidinones 21-26 and 2-{(coumarin-4-oxy)methyl}-4-acetyl-5-substituted-delta2-1,3,4-oxadiazolines 27-33 was performed by the reaction of the hydrazones 3, 4, 7, 9, 12, 14 with mercaptoacetic acid and the hydrazones 3, 4, 5, 7, 12, 15, 16 with acetic anhydride, respectively . The antiviral activities, cytotoxicities and structure-activity relationship (SAR) towards different microorganisms of the prepared compounds were studied.

Adv Microb Physiol, 2003, 47, 255 - 310
Microbial globins; Wu G et al.; Globins are an ancient and diverse superfamily of proteins . The globins of microorganisms were relatively ignored for many decades after their discovery by Warburg in the 1930s and rediscovery by Keilin in the 1950s . The relatively recent focus on them has been fuelled by recognition of their structural diversity and fine-tuning to fulfill (probably) discrete functions but particularly by the finding that a major role of certain globins is in protection from the stresses caused by exposure to nitric oxide (NO)--itself a molecule that has attracted intense curiosity recently . At least three classes of microbial globin are recognised, all having features of the classical globin protein fold . The first class is typified by the myoglobin-like haemprotein Vgb from the bacterium Vitreoscilla, which has attracted considerable attention because of its ability to improve growth and metabolism for biotechnological gain in a variety of host cells, even though its physiological function is not fully understood . The truncated globins are widely distributed in bacteria, microbial eukaryotes as well as plants and are characterised by being 20-40 residues shorter than Vgb . The polypeptide is folded into a two-over-two helical structure while retaining the essential features of the globin superfamily . Roles in oxygen and NO metabolism have been proposed . The third and best understood class comprises the flavohaemoglobins, which were first discovered and partly characterised in yeast . These are distinguished by the presence of an additional domain with binding sites for FAD and NAD(P)H . Widely distributed in bacteria, these proteins undoubtedly confer protection from NO and nitrosative stresses, probably by direct consumption of NO . However, a bewildering array of enzymatic capabilities and the presence of an active site in the haem pocket reminiscent of peroxidases hint at other functions . A full understanding of microbial globins promises advances in controlling the interactions of pathogenic bacteria with their animal and plant hosts, and manipulations of microbial oxygen transfer with biotechnological applications.

Am J Clin Pathol, 2003 Oct, 120(4), 567 - 74
Immunostaining of visceral leishmaniasis caused by Leishmania infantum using monoclonal antibody (19-11) to the Leishmania homologue of receptors for activated C-kinase; Hofman V et al.; It sometimes is difficult to diagnose leishmaniasis in tissue sections or in smears, particularly in unusual sites or if few parasites are present in the lesion . Leishmania species must be differentiated morphologically from a variety of other microorganisms, including Toxoplasma gondii, Histoplasma capsulatum, Trypanosoma cruzi, and Penicillium marneffei . We tested the value of monoclonal antibody p19-11 raised against the Leishmania homologue of receptors for activated C-kinase (LACK) as an immunohistochemical marker for amastigotes of Leishmania infantum . We evaluated a total of 117 paraffin-embedded lesions due to L infantum (92 cases), T gondii (15 cases), H capsulatum (5 cases), T cruzi (3 cases), and P marneffei (2 cases) . Amastigotes of Leishmania species were detected in 92 (100%) of the leishmaniasis lesions . There were no false-positive LACK immunoreactions in any of the toxoplasmosis, histoplasmosis, trypanosomiasis, or penicilliosis specimens (0/25) . We found the anti-LACK antibody p19-11 to be a highly specific and sensitive paraffin-reactive immunohistochemical marker for the confirmation or identification of Leishmania species in tissue sections.

J Pediatr (Rio J), 2003 Sep-Oct, 79(5), 467 - 70
{Chronic recurrent multifocal osteomyelitis of the mandible: report of three cases}; Paim LB et al.; OBJECTIVE: To report three cases of chronic recurrent multifocal osteomyelitis of the mandible, an inflammatory disease affecting one or more bones with absence of isolated microorganisms in affected areas . DESCRIPTION: The first case is a 13 year-old female presenting with pain and fever after dental treatment . The patient received antibiotic treatment for osteomyelitis, but developed progressive enlargement of the mandible and palmoplantar pustulosis . Bone scintigraphy showed intense and diffuse uptake in the mandible . The swelling decreased after indomethacin and hyperbaric oxygen therapy . Case 2 is a 9 year-old female patient with recurrent pain and edema of the right mandible for three years . The diagnosis of osteomyelitis was established and amoxicillin introduced . After three months, tomography showed diffuse mandible osteolysis . Indomethacin and hyperbaric oxygen therapy were introduced, however the patient presented a relapse and was treated with prednisone, rofecoxib and methotrexate . Patient 3, a 10 year-old male, had palmoplantar pustulosis and recurrent enlargement of the mandible . Tomography showed diffuse mandible osteolysis and scintigraphy revealed intense and diffuse uptake in the mandible . The patient was treated with prednisone . Rofecoxib was replaced after two relapses . COMMENTS: Chronic recurrent multifocal osteomyelitis of the mandible is often associated with prolonged pain periods and periods of activity and remission of the inflammatory process . Its recognition is important to prevent the patient from being submitted to prolonged antibiotic therapy and unnecessary invasive procedures.

Clin Microbiol Rev, 2003 Oct, 16(4), 658 - 72
Potential uses of probiotics in clinical practice; Reid G et al.; Probiotics are defined as live microorganisms that, when administered in adequate amounts, confer a health benefit on the host . There is now mounting evidence that selected probiotic strains can provide health benefits to their human hosts . Numerous clinical trials show that certain strains can improve the outcome of intestinal infections by reducing the duration of diarrhea . Further investigations have shown benefits in reducing the recurrence of urogenital infections in women, while promising studies in cancer and allergies require research into the mechanisms of activity for particular strains and better-designed trials . At present, only a small percentage of physicians either know of probiotics or understand their potential applicability to patient care . Thus, probiotics are not yet part of the clinical arsenal for prevention and treatment of disease or maintenance of health . The establishment of accepted standards and guidelines, proposed by the Food and Agriculture Organization of the United Nations and the World Health Organization, represents a key step in ensuring that reliable products with suitable, informative health claims become available . Based upon the evidence to date, future advances with single- and multiple-strain therapies are on the horizon for the management of a number of debilitating and even fatal conditions.

Immunol Lett, 2003 Oct 31, 89(2-3), 237 - 41
Current issues in antigen presentation--focus on the dendritic cell; Chain BM; A personal view of some of the major current issues in antigen presentation, which focuses on the biology of the dendritic cell . The question of dendritic cell heterogeneity is discussed and the inclusion of the "plasmacytoid" dendritic cell in this family is questioned . Defining the molecules involved in dendrite formation may ultimately provide the only reliable signature of the dendritic cell . The role of the dendritic cell as a key link between innate and adaptive immunity is here revisited, and the direct and indirect interactions between dendritic cells and microorganisms are discussed . Reactive oxygen species may be key modulators of dendritic cell function, although it remains unclear whether direct or indirect mechanisms are more important . Study of the cell biology of antigen processing reveals a tightly regulated complex system involving multiple proteinases, adapted to the most efficient presentation of invading microorganisms and playing a key role in shaping the T cell repertoire.

Vet Res, 2003 Sep-Oct, 34(5), 647 - 70
The complement in milk and defense of the bovine mammary gland against infections; Rainard P; The mammary gland of dairy cows, which is prone to infection by various bacteria, mobilizes local and systemic immune defenses to cope with pathogens . The complement system plays an important part in the innate immunity against microorganisms through its bactericidal, opsonic, and phlogistic functions . The amount of the complement in the milk of healthy glands of dairy cows is low . Moreover, the classical pathway of activation is not functional because of a shortage in C1q . By contrast, the alternative pathway is active, deposits C3b and C3bi on bacteria, and generates amounts of C5a which are highly variable among cows . A slight inhibition of the bactericidal/hemolytic activities, of the deposition of C1q on bacteria, and of the phlogistic activity of C5a makes milk a rather anti-inflammatory fluid . The inhibitory activity does not involve C3b/ C3bi deposition on bacteria, nor the generation of C5a by the alternative pathway . When inflammation develops, the blood-derived complement components overcome the inhibitions and complement-dependent bactericidal, opsonic and phlogistic activities may be high in milk . Further research is necessary to evaluate the contribution of C5a to the recruitment of leukocytes in the mammary gland, and to specify the links between the complement system and the response of resident cells (leukocytes and mammary epithelial cells) to infection stimulus . This will help to define the contribution of the complement system to resistance against mastitis, and could help to differentiate animals more or less resistant to this frequent and costly disease.

Infection, 2003 Oct, 31(5), 341 - 9
Clinical significance of human intestinal spirochetosis--a morphologic approach; Korner M et al.; Intestinal spirochetosis (IS) is a condition defined morphologically by the presence of spirochetal microorganisms attached to the apical cell membrane of the colonic and rectal epithelium . Intestinal spirochetes comprise a heterogeneous group of bacteria . In humans Brachyspira aalborgi and Brachyspira pilosicoli predominate . Prevalence rates of IS are low where living standards are high, in contrast to poorly developed areas where IS is common . Homosexuals and HIV-infected individuals are at high risk of being colonized . Clinical significance in individual cases has remained unclear up to now . A review of the literature reveals that invasion of spirochetes beyond the surface epithelium is associated with gastrointestinal symptoms which respond to antibiotic treatment (metronidazole), whereas patients lacking this feature are mostly asymptomatic . Homosexual and HIV-positive men are more likely to be symptomatic irrespective of invasion . Rare cases of spirochetemia and multiple organ failure have been reported in critically ill patients.

Adv Space Res, 2003, 31(7), 1731 - 5
Increase of atmospheric CO2: response patterns of a simple terrestrial man-made ecosystem; Somova LA et al.; Simple models of terrestrial ecosystems with a limited number of components are an efficient tool to study the main laws of functioning of populations, including microbial ones, and their communities, as components of natural ecosystems, under variable environmental conditions . Among other factors are the increase of carbon dioxide in the atmosphere and limitation of plants' growth by biogenic elements . The main types of ecosystems' responses to changes in environmental conditions (a change in CO2 concentration) have been demonstrated in a "plants-rhizospheric microorganisms-artificial soil" simple experimental system . The mathematical model of interactions between plants and microorganisms under normal and elevated atmospheric CO2 and limitation by nutrients (nitrogen and phosphorus) yielded a qualitative agreement between calculated and experimental values of limiting substances concentrations and release rates of exudates . c2003 COSPAR . Published by Elsevier Science Ltd . All rights reserved.

FEMS Microbiol Rev, 2003 Oct, 27(4), 493 - 504
Adaptation of Borrelia burgdorferi in the tick and the mammalian host; Anguita J et al.; Borrelia burgdorferi, the causative agent of Lyme disease, shows a great ability to adapt to different environments, including the arthropod vector, and the mammalian host . The success of these microorganisms to survive in nature and complete their enzootic cycle depends on the regulation of genes that are essential to their survival in the different environments . This review describes the current knowledge of gene expression by B . burgdorferi in the tick and the mammalian host . The functions of the differentially regulated gene products as well as the factors that influence their expression are discussed . A thorough understanding of the changes in gene expression and the function of the differentially expressed antigens during the life cycle of the spirochete will allow a better control of this prevalent infection and the design of new, second generation vaccines to prevent infection with the spirochete.

Biotechnol Adv, 1988, 6(4), 683 - 95
Microbial DNA diagnostic technology; Pasternak JJ; The principle objectives when creating a robust DNA diagnostic assay system are sensitivity, specificity and minimal read-time . To meet these ends, depending on the specifically defined test, various aspects of molecular hybridization methodology must be optimized . In particular, among other things, attention has focused on (i) formulating highly specific probes; (ii) devising sensitive nonisotopic detection systems, (iii) minimizing the extent of preparing clinical samples for assaying, (iv) amplifying the target sequence to augment sensitivity and (v) enhancing hybridization kinetics to speed up the reaction period . In this article, some recent studies that are directed to the development of nucleic acid hybridization systems for clinical diagnosis of microorganisms are considered.

Biotechnol Adv, 1990, 8(3), 539 - 46
Microbial interactions with germanium; Lee H et al.; Germanium is a semi-precious, widely-dispersed, and biologically non-essential metal with considerable potential for application in the fields of electronics, computer engineering and medicine . Research on the physiology and genetics of germanium toxicity and accumulation in microorganisms has received scant attention . An understanding of these aspects is important from both fundamental and applied points of view . This review will examine the diverse range of interactions between germanium and microbial species.

Biotechnol Adv, 1991, 9(2), 241 - 52
Applications of microbial surfactants; Van Dyke MI et al.; Biosurfactants are amphiphilic compounds produced by microorganisms that are capable of decreasing surface and interfacial tensions . They are useful in remediation of insoluble organic pollutants in soil and marine environments . There are also a large number of industrial uses for biosurfactants . This paper reviews recent research on applications of microbially-produced surfactants.

Biotechnol Adv, 1994, 12(3), 539 - 55
Surface display of antibodies; Little M et al.; To screen antibody libraries that contain many millions of different clones, a selection system is required with an efficiency comparable to that of the immune system . This can be achieved by displaying antibodies on the surface of microorganisms containing the antibody's gene, analogous to the expression of the IgM antigen receptor on the surface of unactivated B-lymphocytes . Specific clones can then be selected using immobilized antigens . The minor coat protein of filamentous phages, pIII, which initiates the infection of E.coli by binding to their F-pili, and the major coat protein, pVIII, have been used as carriers for displaying antibodies on the phage surface . Recombinant antibodies have also been targeted to the cell surface of bacteria by fusing them with outer membrane components derived from lipoproteins, OmpA and an IgA protease . However, only the pIII system has been routinely used for screening antibody libraries . Here we describe the various antibody surface display systems and the screening of antibody libraries generated from the gene repertoire of lymphocytes and by gene synthesis . Finally, we have made a short comparison of the bacterial production of Fabs versus single chain antibodies (scFv).

Biotechnol Adv, 1994, 12(4), 687 - 92
Live cultures: their use in industrial biotechnology; Power R et al.; Advances in the areas of molecular genetics and molecular biology provide a tremendous opportunity for development of custom-made microorganisms for human and animal use . This reviews discusses the current and future potentials of these organisms for use in the food and agricultural industries.

Biotechnol Adv, 1990, 8(1), 291 - 301
Achievements in microbial technology; Demain AL; In 1973, recombinant DNA technology was born and the age of the "new biotechnology" came upon us . Today we are seeing the amazing results of recombinant DNA technology, hybridoma technology, enzyme engineering and protein engineering . These techniques are exerting major effects on basic research and on health care, diagnostics and agriculture and soon will bring about changes in other industries such as petroleum, mining, foods and chemicals . Entire pathways of primary and secondary metabolism have been cloned and expressed in foreign microorganisms . The development of recombinant DNA technology is having its major impact on the production of rare polypeptides such as mammalian enzymes, hormones, antibodies and biological response modifiers . In addition to natural polypeptides, analogs are being produced by recombinant DNA technology and this has added an extra dimension of excitement to the field . The future is thus insured for the expanded use of microorganisms in the biotechnological world and the continued improvement in microbial processes to reduce the cost of drugs, enzymes and specialty chemicals.

Biotechnol Adv, 1990, 8(1), 277 - 90
Entropy balances of microbial product formation; Bormann EJ et al.; Considering the approach of Bermudez and Wagensberg (1986) devoted to the entropy balance of growing microorganisms some equations were developed which describe particularly the entropy balance of microbial product formation . The formula allows to determine the coefficients of resistance R(mn) and of coupling L(mn) according to rates of growth, product formation, maintenance metabolism and heat evolution assuming a linear relationship between thermodynamic fluxes and forces.In order to check the usefulness of the derived model appropriate experimental data of two microbial batch processes concerning production of L-lysine and the antibiotic nourseothricine were taken into account . The results showed similar courses of entropy balances despite different pathways of product formation which were characterized by an overshoot of entropy production at the beginning of biosynthesis of the primary and secondary metabolite . This fact was interpreted as a more general phenomenon for microorganisms under inbalanced nutritional conditions.

Biotechnol Adv, 1990, 8(1), 1 - 27
Overproduction of microbial products--facts and ideas; Vanek Z et al.; Overproduction of microbial metabolites is related to developmental phases of microorganisms . Inducers, effectors, inhibitors and various signal molecules play a role in different types of overproduction . Primary and secondary metabolism are interconnected . Biosynthesis of enzymes catalyzing metabolic reactions in microbial cells is controlled by well-known positive and negative mechanisms, e.g . induction, repression, catabolite repression, mechanisms controlling enzyme activity include isosteric and allosteric interactions, e.g . competitive and non-competitive inhibition, allosteric effects, molecular conversion etc . Biosynthesis of secondary metabolites is catalyzed by unaltered enzymes of primary metabolism, by altered enzymes of primary metabolism and by specific enzymes of secondary metabolism . In addition to classical mutagenesis and selection of suitable microbial cells, methods of molecular genetics are used in the overproduction of microbial products.

Biotechnol Adv, 1994, 12(2), 357 - 91
Advances in microbial biotechnology of bile acids; Mahato SB et al.; The recent advances in microbial biotechnology of production of bile acid metabolites helped to identify a number of neutral and acidic steroidal compounds useful as drugs and drug intermediates on a scale which would not have been possible by classical chemical transformations . Microbial transformations viz., hydroxylation, dehydroxylation, reduction of the carbonyl moieties, epimerization, side-chain metabolism, introduction of carbon-carbon double bonds into the steroid nucleus, deconjugation of bile acid conjugates carried out by various microorganisms for production of useful metabolites with special reference to newer techniques including cell immobilization and transposon mutagenesis for selective transformations are reviewed . The different pathways of microbial degradation of bile acids leading to the formation of various products are discussed . A compilation of the metabolites formed by various microorganisms from the bile acids or their conjugates and reported during the period 1979-1992 is also provided.

Biotechnol Adv, 1992, 10(2), 179 - 89
High-level expression of foreign genes in Hansenula polymorpha; Gellissen G et al.; The methylotrophic yeast Hansenula polymorpha belongs to a limited number of non-Saccharomyces yeast species used as hosts for heterologous gene expression . It has successfully been applied for the production of hormones, antigens and enzymes . The system excells by mitotically stable recombinant strains, high productivity and faithful processing of the produced polypeptides . The favourable characteristics of this microorganism for protein production at an industrial scale are described in the following article focusing on some recent representative examples.

Biotechnol Adv, 1984, 2(1), 101 - 20
Mycorrhizae and their potential use in the agricultural and forestry industries; Peterson RL et al.; Mycorrhizal fungi associated with plant roots increase the absorption of nutrients, particularly phosphorus, and thus enhance the growth of crop plants and trees . Vesicular-arbuscular mycorrhizae (VAM) occur in approximately 90% of all vascular plants including most of the important agricultural species, whereas ectomycorrhizae are found in most of the economically important tree species of the temperate regions of the world, and in some tropical trees . These symbiotic associations are, therefore, important in crop and biomass production . For this reason they are receiving considerable attention in agriculture and forestry . Currently, VAM are utilized in fumigated soils, greenhouse crops, and in the reclamation of disturbed sites . Ectomycorrhizae are employed in the establishment of trees in nurseries, in reforestation programs, and in the production of containerized seedlings . Production of VAM and ectomycorrhiza inoculum for large scale projects is now feasible but many basic questions related to persistence of these fungi in field situations, competition with other microorganisms, and particularly the most efficient fungi to use for particular hosts remain largely unanswered.

Biotechnol Adv, 1984, 2(1), 43 - 62
Pretreatment for enhanced hydrolysis of cellulosic biomass; Grethlein HE; This review will cover a number of physical and chemical pretreatment methods for cellulosic substrates which enhance their hydrolysis by cellulase or consumption by microorganisms . While the emphasis is on the literature of the last two years, some earlier work is cited which has influenced the work in the pretreatment field . In order to interpret the effects of a pretreatment method, emphasis in the past has been on crystallinity index (CI) and lignin content . Although these parameters happen often to correlate with the rate or extent of hydrolysis, it is suggested that a more basic parameter is the pore size distribution of the wet substrate and the associated surface area available to the cellulase that is the major factor in determining the effectiveness of a pretreatment method.

Biotechnol Adv, 1984, 2(1), 35 - 42
Continuous cultivation of microorganisms; Sikyta B; This review presents selected studies on continuous cultivation including the genetics and selection of microorganisms, production of biomass, liquid fuels and other products, published in 1982-1984 and bearing a relationship to biotechnology.

Biotechnol Adv, 1983, 1(2), 279 - 88
Strategies for cultivation of recombinant organisms; Imanaka T; An overview on the strategies for cultivation of recombinant organisms is presented in three sections, that is, the stability of plasmids, expression of cloned genes and an example of a genetically engineered microorganism.

Biotechnol Adv, 1997, 15(3-4), 583 - 620
Cellulose degrading enzymes and their potential industrial applications; Bhat MK et al.; Bioconversion of cellulose to soluble sugars and glucose is catalyzed by a group of enzymes called cellulases . Microorganisms including fungi, bacteria and actinomycetes produce mainly three types of cellulase components--endo-1,4-beta-D-glucanase, exo-1,4-beta-D-glucanase and beta-glucosidase--either separately or in the form of a complex . Over the last several decades, cellulases have become better understood at a fundamental level; nevertheless, much remains to be learnt . The tremendous commercial potential of cellulases in a variety of applications remains the driving force for research in this area . This review summarizes the present state of knowledge on microbial cellulases and their applications.

Biotechnol Adv, 1999 Dec 15, 17(7), 561 - 94
Microbial alkaline proteases: from a bioindustrial viewpoint; Kumar CG et al.; Alkaline proteases are of considerable interest in view of their activity and stability at alkaline pH . This review describes the proteases that can resist extreme alkaline environments produced by a wide range of alkalophilic microorganisms . Different isolation methods are discussed which enable the screening and selection of promising organisms for industrial production . Further, strain improvement using mutagenesis and/or recombinant DNA technology can be applied to augment the efficiency of the producer strain to a commercial status . The various nutritional and environmental parameters affecting the production of alkaline proteases are delineated . The purification and properties of these proteases is discussed, and the use of alkaline proteases in diverse industrial applications is highlighted.

Biotechnol Adv, 2001 Apr 1, 19(2), 119 - 32
Microbial leaching of metals from sulfide minerals; Suzuki I; Microorganisms are important in metal recovery from ores, particularly sulfide ores . Copper, zinc, gold, etc . can be recovered from sulfide ores by microbial leaching . Mineral solubilization is achieved both by 'direct (contact) leaching' by bacteria and by 'indirect leaching' by ferric iron (Fe(3+)) that is regenerated from ferrous iron (Fe(2+)) by bacterial oxidation . Thiobacillus ferrooxidans is the most studied organism in microbial leaching, but other iron- or sulfide/sulfur-oxidizing bacteria as well as archaea are potential microbial agents for metal leaching at high temperature or low pH environment . Oxidation of iron or sulfur can be selectively controlled leading to solubilization of desired metals leaving undesired metals (e.g., Fe) behind . Microbial contribution is obvious even in electrochemistry of galvanic interactions between minerals.

Biotechnol Adv, 2001 Sep, 19(5), 387 - 97
Biotechnological production of prostaglandin; Yilmaz E; Prostaglandins (PGs) are the oxidation products of PG endoperoxide (PGH) synthase and other tissue enzymes . They occur in a tissue-specific manner and act as local hormones . Biotechnological production of PGs has been of interest, but not yet fully established . Biological tissues have been used as PG sources, but this disturbs ecological balance, and the cost of production is very high for commercial purposes . On the other hand, various microorganisms have been shown to synthesize them de novo, or biotransform precursors to active molecules, but these processes have not been further evaluated . Using mammalian enzymes in free or immobilized form is a promising new approach to synthesize PG from fatty acid substrates . Rapid enzyme inactivation during the catalysis is the main problem to be solved . Optimization of factors in the reactions and the design of special reactors that will allow removal of products continuously from the reaction medium without affecting enzyme activity need immediate attention from researchers and the pharmaceutical industry.

Biotechnol Adv, 2001 Sep, 19(5), 371 - 85
Extracellular biopolymeric flocculants . Recent trends and biotechnological importance; Salehizadeh H et al.; Many microorganisms secrete extracellular biopolymeric flocculants (EBFs) in the culture broth . This work reviews the development of EBF research and applications . Aspects discussed include a comparison of the chemical and biological flocculating agents, isolation of EBF-producing microorganisms, culture conditions, mechanisms of flocculation, the chemical structure of EBFs, and the role of physicochemical factors in the flocculating activity.

Biotechnol Adv, 1995, 13(1), 75 - 90
The use of immunological methods to detect and identify bacteria in the environment; Schloter M et al.; Immunological detection methods have become increasingly important in microbial ecology for the tracking of specific microorganisms and for community analysis . For a reliable application of these techniques, the monoclonal antibodies or polyclonal antisera used have to fulfill several quality criteria . Cross reactivity, cellular localization of the antigenic determinant, affinity characteristics and the expression of the antigenic determinant at environmental conditions have to be determined . Immunological methods can be used for the identification, quantification and enrichment of specific bacteria in extracts as well as for the visualization of cells in situ . The sensitivity of advanced immunological methods can be compared to PCR techniques . Using image processing of epifluorescence micrographs or confocal laser scanning microscopy, the immunofluorescence approach can now be applied to study complex environmental samples.

Biotechnol Adv, 1996, 14(1), 1 - 15
Multifactional glucanases; Kumar NN et al.; Diverse functional roles of multifunctional proteins arise from either their independent functional domains or dual activities mediated through a single active site . Presence of multifunctional proteins significantly enhances the metabolic efficiency of a cell . Microorganisms utilising complex substrates with extensive microheterogeneities, such as carbohydrates evolved batteries of multifunctional glucanases, facilitating parsimonious utilisation of these substrates . Various attempts have since been made to artificially construct these glucanases . Analysis of information on various glucanases would be helpful in understanding the evolutionary interrelationship between this class of enzymes and will give an insight into the structural features controlling different unrelated activities . This review examines the genesis, evolution and structural features of multifunctional glucanases.

Biotechnol Adv, 1995, 13(3), 491 - 551
Process-scale disruption of microorganisms; Middelberg AP; Common hosts for the large-scale manufacture of biological products, such as Escherichia coli and Saccharomyces cerevisiae, do not excrete products to the medium . Effective techniques for cell disruption are therefore required . These include physical, chemical, enzymatic and mechanical methods . Mechanical methods such as bead milling, high-pressure homogenization, and microfluidization are preferred . However, gentler, specific methods are receiving increasing attention particularly when used in combination to synergistically exploit their different specificities . Benefits can also be derived by integrating product release and recovery . In all cases it is essential to consider the interaction of the disruption operation with downstream units and to clearly demonstrate the cost benefits of alternative strategies.

Helicobacter, 2003, 8(5), 568 - 72
Placental acquisition of maternal specific IgG and Helicobacter pylori colonization in infancy; Bunn JE et al.; BACKGROUND: Colonization with Helicobacter pylori generally occurs in infancy, and the microorganism is often acquired from close family members . Rate of infant colonization may be affected by maternal immune status . METHODS: To investigate the potential protective effect of anti-H . pylori immunoglobulin G (IgG) acquired via the placenta, 65 mothers and their infants were studied from the infant's birth for 1 year . Circulating IgG antibodies were measured by enzyme-linked immunosorbent assay (ELISA) in cord blood and every 8 weeks . Immunoblotting was performed on sera from infants with significant increases in IgG levels . Rate of infant H . pylori colonization was measured by 13C urea breath tests every 4 weeks from the age of 12 weeks . RESULTS: Maternal and infant cord blood specific IgG levels were correlated (R2 =.747, p <.001) . Infant H . pylori specific IgG fell 5-fold compared to maternal levels over the first 6 months of life, and rose subsequently in many cases, with the development of novel immunoblot patterns . There were no significant associations between the age at first positive urea breath test and maternal or infant cord specific H . pylori IgG levels . CONCLUSIONS: Transplacentally acquired specific IgG antibody does not protect infants from colonization by H . pylori.

Andrologia, 2003 Oct, 35(5), 258 - 62
The 2001 Giessen Cohort Study on patients with prostatitis syndrome--an evaluation of inflammatory status and search for microorganisms 10 years after a first analysis; Schneider H et al.; During the last years tremendous changes have occurred in the epidemiologic knowledge and the diagnostic process of the prostatitis syndrome . A new worldwide-accepted classification system has become the gold standard in contemporary literature . The aim of this study was to compare the inflammatory and infectious status of men with prostatitis syndrome with results from our study cohort from 1992 . A total of 168 symptomatic men (mean age 43.2 years; range 18-79) attending the Giessen prostatitis outpatient department were included . All men underwent a standard four-glass-test including leucocyte analysis in all specimens . A routine search for Ureaplasma urealyticum and Chlamydia trachomatis was performed . Ejaculate analysis following World Health Organization (WHO) criteria has been performed including the evaluation of increased number of peroxidase-positive leucocytes (PPL) . Men were classified according to the National Institutes of Health (NIH) prostatitis classification . The distribution of patients according to NIH criteria is as follows: NIH II (4.2%), NIH IIIA (31.5%), NIH IIIB (50.0%) and urethroprostatitis (14.3%) . Chlamydial infection was present in one man (0.6%) . Only two men with increased leucocytes in prostatic secretions demonstrated > or =106 million ml-1 PPL in semen . As compared with our cohort study 10 years ago, the proportion of the different subtypes of the prostatitis syndrome have remained stable . The aetiological spectrum of chronic bacterial prostatitis has not changed whereas, in contrast, the prevalence of C . trachomatis now is found to be strikingly reduced . Using the WHO cutpoints for leucocytospermia the inclusion of seminal leucocytes to the diagnostic process has not influenced the distribution between inflammatory (type NIH IIIA) and noninflammatory (type NIH IIIB) chronic pelvic pain syndrome.

J Environ Qual, 2003 Sep-Oct, 32(5), 1735 - 42
Competitive degradation between the fumigants chloropicrin and 1,3-dichloropropene in unamended and amended soils; Zheng W et al.; The mixture of 1,3-dichloropropene (1,3-D) and chloropicrin (CP) is used as a preplant soil fumigant . In comparison with individual fumigants, application of a mixture may affect the environmental dissipation and fate of each chemical, such as emission and degradation . We investigated the degradation of CP, 1,3-D, and their mixture in fresh soils and sterile soils, and evaluated the competitive characteristic of fumigants in the mixture . The degradation of low concentrations of CP in fresh soil was accelerated at early times in the presence of 1,3-D, whereas the addition of CP reduced the degradation rate of trans-1,3-D, possibly by inhibiting the activity of trans-1,3-D degrading microorganisms . The potential of applying amendments to the soil to increase the rate of CP and 1,3-D degradation was also illustrated . The degradation of both fumigants was significantly enhanced in soils amended with ammonium thiosulfate (ATS) and sodium diethyldithiocarbamate (Na-DEDTC) compared with unamended soil . Competitive degradation was observed for CP in amended soils in the presence of 1,3-D . The degradation of cis-1,3-D in amended soils spiked as a mixture of 1,3-D and CP was repressed compared with the rate of degradation in samples spiked with 1,3-D only . This implied that in abiotic degradation, CP and cis-1,3-D competed for a limited number of reaction sites in amended soil, resulting in decreased degradation rates . No significant influence of fumigant mixtures was observed for trans-1,3-D in amended soil.

Bull Group Int Rech Sci Stomatol Odontol, 2003 Jan-Apr, 45(1), 29 - 33
Study on the occurrence of microorganisms on the post-surgical maxillary prostheses with obturators and in the post-surgical cavities of maxilla; Wieckiewicz W et al.; The bacterial environment of the mouth cavity may be subjected to change under influence of various factors, such as surgical removal of neoplasm tumors and in consequence the wearing of post-surgical prostheses with obturators . The purpose of the paper was to study the conceivable differences in occurrence of particular types of microorganisms found on the margin of post-surgical cavities and on the prosthetic obturators . The performed microbiologic examinations revealed that more pathologic bacterial flora was found on the obturators than in the post-surgical cavities . The authors conclude that the post-surgical patients should pay more attention to the very accurate hygiene of their prostheses and the mouth cavity as well.

Oncol Rep, 2003 Nov-Dec, 10(6), 1851 - 5
Induction of tumor cell death by high hydrostatic pressure as a novel supporting technique in orthopedic surgery; Diehl P et al.; As vegetative forms of microorganisms are impaired by high hydrostatic pressure (HHP) in the range of 400-600 MPa, the non-thermal inactivation of vegetative bacteria, yeasts, and moulds present in foods such as jams, fruit juices, and dressings by HHP is now well-established . Eukaryotic cells, when subjected to HHP are also damaged . In the present study, the effect of HHP on cell viability of human osteoblasts (HOB), human fibroblasts (HFB), and different tumor cell lines such as osteosarcoma cells SAOS-2, human histiocytic leukemia cells U-937, and the ovarian cancer cell line OV-MZ-6 was investigated . Therefore the different cell lines were subjected to pressures between 50 and 400 MPa and tested for viability . At HHP of 100 MPa (10 min) about 80% of the various cell lines were still alive . At 350 MPa all of the cells were damaged and dead . The three tumor cell lines investigated were slightly more resistant to HHP (50% dead at 170-193 MPa) than HOB and HFB (50% dead at 130-145 MPa) . The present study demonstrates that both normal cells and tumor cells are rapidly inactivated by HHP treatment.

Biotech Histochem, 2003 Apr, 78(2), 71 - 4
Sieve element specific labeling with an anti-idiotypic monoclonal antibody mimic of the phytotoxin dothistromin; Sutherland PW et al.; A monoclonal antibody, 12C9, an anti-idiotypic mimic of dothistromin, a toxin produced by Dothistroma pini, was found to label the cell wall of sieve elements in a number of different plant tissues and species . The antibody labeled apple leaf tissue, tobacco leaf mid vein, leaf and meristem, and Coprosma robusta leaf mid vein . Labeling was restricted to cell walls of sieve elements and did not label the companion cells or the lumen of the cells . The antibody labeled over a wide range of dilutions . This antibody could be used to differentiate sieve elements from other types of phloem . It could also be used to co-localize sieve elements and microorganisms such as phytoplasmas stained with DAPI.

Mol Cell Proteomics, 2003 Dec, 2(12), 1284 - 1296 Epub 2003 Oct 6.
Comprehensive Proteomic Profiling of the Membrane Constituents of a Mycobacterium tuberculosis Strain; Gu S et al.; Mycobacterium tuberculosis is an infectious microorganism that causes human tuberculosis . The cell membranes of pathogens are known to be rich in possible diagnostic and therapeutic protein targets . To compliment the M . tuberculosis genome, we have profiled the membrane protein fraction of the M . tuberculosis H37Rv strain using an analytical platform that couples one-dimensional SDS gels to a microcapillary liquid chromatography-nanospray-tandem mass spectrometer . As a result, 739 proteins have been identified by two or more distinct peptide sequences and have been characterized . Interestingly, approximately 450 proteins represent novel identifications, 79 of which are membrane proteins and more than 100 of which are membrane-associated proteins . The physicochemical properties of the identified proteins were studied in detail, and then biological functions were obtained by sorting them according to Sanger Institute gene function category . Many membrane proteins were found to be involved in the cell envelope, and those proteins with energy metabolic functions were also identified in this study.

Appl Environ Microbiol, 2003 Oct, 69(10), 6165 - 73
Immobilization of arsenite and ferric iron by Acidithiobacillus ferrooxidans and its relevance to acid mine drainage; Duquesne K et al.; Weathering of the As-rich pyrite-rich tailings of the abandoned mining site of Carnoules (southeastern France) results in the formation of acid waters heavily loaded with arsenic . Dissolved arsenic present in the seepage waters precipitates within a few meters from the bottom of the tailing dam in the presence of microorganisms . An Acidithiobacillus ferrooxidans strain, referred to as CC1, was isolated from the effluents . This strain was able to remove arsenic from a defined synthetic medium only when grown on ferrous iron . This A . ferrooxidans strain did not oxidize arsenite to arsenate directly or indirectly . Strain CC1 precipitated arsenic unexpectedly as arsenite but not arsenate, with ferric iron produced by its energy metabolism . Furthermore, arsenite was almost not found adsorbed on jarosite but associated with a poorly ordered schwertmannite . Arsenate is known to efficiently precipitate with ferric iron and sulfate in the form of more or less ordered schwertmannite, depending on the sulfur-to-arsenic ratio . Our data demonstrate that the coprecipitation of arsenite with schwertmannite also appears as a potential mechanism of arsenite removal in heavily contaminated acid waters . The removal of arsenite by coprecipitation with ferric iron appears to be a common property of the A . ferrooxidans species, as such a feature was observed with one private and three collection strains, one of which was the type strain.

Cell Microbiol, 2003 Nov, 5(11), 743 - 54
Invasion and survival strategies of Anaplasma phagocytophilum; Carlyon JA et al.; Anaplasma phagocytophilum is an aetiological agent of human granulocytic ehrlichiosis, an emerging tick-borne zoonosis in the United States and Europe . This obligate intracellular bacterium is unique in that it colonizes polymorphonuclear leucocytes (neutrophils) . Neutrophils are key players in innate immunity . These short-lived phagocytes ingest invading microorganisms and destroy them by various means, which include fusing the bacteria-containing phagosome with acidic lysosomes as well as directing toxic oxidative and proteolytic compounds into the phagosomal lumen . Its tropism for neutrophils indicates that A . phagocytophilum uses strategies for evading and/or neutralizing these microbicidal activities . This review focuses on some of the mechanisms that A . phagocytophilum uses for neutrophil adhesion, surviving within the hostile intracellular environment of its host neutrophil and for effectively disseminating to naive host cells.

Rev Environ Health, 2003 Apr-Jun, 18(2), 131 - 51
Molecular microbial biodiversity assessment: a biological indicator of soil health; Alkorta I et al.; The soil performs a variety of key functions: (i) provides the food, fuel, and fiber needs of the world's population, (ii) regulates the quality of the air and water, (iii) decomposes organic wastes, (iv) recycles nutrients, and (v) acts as a sink for pollutants (including global gases) . Soil degradation is increasingly recognized as an urgent environmental issue and a crucial need exists for the capacity to evaluate soil health . After all, soil health is a key component in sustaining the different world's ecosystems and the myriad of natural and socioeconomic systems they support . In this respect, soil health has been defined as the "continued capacity of a specific kind of soil to function as a vital living system, within natural or managed ecosystem boundaries, to sustain plant and animal productivity, to maintain or enhance the quality of air and water environments, and to support human health and habitation" . A biological indicator is an organism, a part of an organism, or a community of organisms, used to obtain information about environmental quality . The assessment of microbial biodiversity has the potential to provide useful insight into the health and functioning of soil . Our inability to culture most microorganisms that are present in soils has, until recently, impaired studying the relationships between the structure and the function of soil microbial communities . This shortcoming has been recently overcome by using several molecular techniques that allow the detection, enumeration, and characterization of soil microorganisms without cultivation.

Water Sci Technol, 2003, 48(4), 163 - 8
Membrane separation to improve degradation of road side grass by rumen enhanced solid incubation; Dalhoff R et al.; Membrane separation proved to be an excellent means to maintain high residence time of microorganisms in an anaerobic hydrolysis reactor, and relatively low concentration of hydrolysis products . The microbial biocommunity typical for the rumen environment could be maintained, and the reactor efficiency of the reactor improved . Less than 4 days were reqired to reach almost complete hydrolysis of the grass fed into the reactor . To avoid blocking of the membrane unit, a backwash system is necessary . The membranes needed to be backwashed every 20 min with 4 bar gas-pressure for 10 s . After this treatment the initial permeability was regained . The plant was operated with a flux of 12 ml h(-1) cm(-2) on average . The transmembrane pressure was in the range of 0.8-0.9 bar . 90% of the dissolved fatty acids permeated through the membrane.

Water Sci Technol, 2003, 48(4), 105 - 12
Investigation of variations in microbial diversity in anaerobic reactors treating landfill leachate; Calli B et al.; A young landfill leachate containing elevated total ammonia concentrations frequently as high as 2,700 mg/l was anaerobically treated for 1,015 days in two different laboratory-scale anaerobic reactors configured as sludge blanket and hybrid bed . In this paper, the last 265 days of this long-term anaerobic treatability study are presented . Effects of high ammonia concentrations on reactor performances were correlated to the variations in microbial diversity by identifying the dominant microorganisms with FISH (fluorescent in-situ hybridization), cloning, DGGE (denaturing gradient gel electrophoresis) and morphological analysis . The results have indicated that the high ammonia landfill leachate can be treated successfully by using either an UASB or a hybrid bed reactor if temporary pH adjustments in the reactor influents are made when high ammonia concentrations are experienced . Consequently, COD removal efficiency is independent of microbial diversity and reactor configuration and it depends on the biodegradable portion of the leachate . Under this circumstance, stability of the reactors with low levels of acetate was supported by the abundance of Methanosaeta population . In both of the reactors, some populations of Methanobacteriaceae were also detected while other methanogenic species were virtually absent . However, after the termination of pH adjustment at Day 860, reactors immediately became unstable due to the sudden increase in free ammonia concentration up to 400 mg/l . COD removal efficiency decreased to 42% in the hybrid bed and to 48% in the UASB reactor . The durations of inhibitions were not long enough to severely deteriorate the massive Methanosaeta cells; therefore, many of them were again identified after two free ammonia inhibitions . However, subsequently, long filamentous morphologies of Methanosaeta cells shifted to shorter filaments and they lost their aggregating property.

Eur Radiol . 2003 Oct 3; {Epub ahead of print}
Osteomyelitis in infants and children; Schmit P et al.; Infection of the pediatric skeletal system is a dramatic diagnostic and therapeutic challenge . The consequences of a missed diagnosis could lead to long-lasting functional sequelae . This disease has multiple aspects according to the clinical presentation and evolution, the causative microorganism, the site of infection, and, finally, the patient's age and immunological status . The choice of the most appropriate imaging modality according to the clinical presentation is very important to assess the diagnosis, the extent of the disease, and to obtain the causative germ when needed; thus, plain films, ultrasound, nuclear medicine, computed tomography, and MR imaging are discussed, each of them having pros and cons . This multimodality approach of the musculoskeletal infections is mandatory to guide surgery, to deliver an appropriate antibiotic therapy, and to reduce the rate of long-term functional sequelae.

Transplant Proc, 2003 Sep, 35(6), 2207 - 8
Oral cavity as a potential source of infections in recipients with diabetes mellitus; Piekarczyk J et al.; Our previous observations showed alterations of oral cavity status among hemodialyzed patients and kidney allograft recipients as well as differences in the prevalence and composition of microorganisms occurring in the mouths of patients . In the present work, we analysed the results of oral cavity examinations, the identification of microorganisms, and the assessment of their importance to kidney allograft recipients or hemodialyzed patients with diabetes mellitus, in comparison with nondiabetic recipients, dialyzed patients, and control patients.

J Invest Surg, 2003 Sep-Oct, 16(5), 275 - 81
Hyperoxic condition prevents bacterial translocation and elevation of plasma microorganism components during hemorrhagic shock; Endo Y et al.; To evaluate the influence of hyperoxic conditions on bacterial translocation (BT) and microorganism components during hemorrhagic shock, rats were divided into a group breathing 100% oxygen and a group breathing room air . The groups were then subjected to hemorrhagic shock . Systemic blood and mesenteric lymph nodes were cultured for BT, and systemic plasma concentrations of microorganism components were measured by the silkworm larvae plasma (SLP) test and the endotoxin test . Hyperoxic conditions prevented both BT and plasma SLP-reactive substance (peptidoglycan and beta-glucan) elevation during hemorrhagic shock . Our findings suggest that hyperoxic treatment might improve host conditions during hemorrhagic shock.

Environ Health Perspect, 2003 Oct, 111(13), 1590 - 4
Antibiotics in dust originating from a pig-fattening farm: a new source of health hazard for farmers?
Hamscher G, Pawelzick HT, Sczesny S, Nau H, Hartung J.
Pig-house dust originates from feed, bedding, feces, and the animals themselves . If the animals receive drugs such as antibiotics, residues of these substances may occur in manure, in the air, or on surfaces of the respective animal house . In a retrospective study, we investigated dust samples collected during two decades from the same piggery for the occurrence of various antibiotics . In 90% of these samples, we detected up to five different antibiotics, including tylosin, various tetracyclines, sulfamethazine, and chloramphenicol, in total amounts up to 12.5 mg/kg dust . High dust exposure in animal confinement buildings is believed to be a respiratory health hazard because of the high content of microorganisms, endotoxins, and allergens . Further risks may arise from the inhalation of dust contaminated with a cocktail of antibiotics . Apart from that, our data provide first evidence for a new route of entry for veterinary drugs in the environment.

Int J Food Microbiol, 2003 Nov 15, 88(1), 85 - 9
Viability of Saccharomyces cerevisiae cells exposed to low-amperage electrolysis as assessed by staining procedure and ATP content; Guillou S et al.; Assessment of yeast viability by plate counts, ATP determination and FUN-1 viability staining was performed to study sublethal injury of Saccharomyces cerevisiae cells submitted to low-amperage electrolysis . Lethal effects of electrolysis were confirmed by all methods, demonstrated by the decrease in viable counts observed during electrolysis . FUN-1 viability staining and ATP determination appeared to demonstrate higher survivors than plate counts . To study possible recovery of certain yeast cells damaged by electrolysis thus rendering them nonculturable, yeast suspensions were stored in phosphate buffer at 4 and 20 degrees C . Increase in viable counts and ATP content of treated yeast cells was observed during storage at 20 degrees C, whereas viable counts of treated and control yeast cells were shown to decrease during storage at 4 degrees C . The increase in the number of viable cells appeared to be the result of repair of damaged cells rather than regrowth of few cells remaining culturable . The lethal efficacy of electrolysis might be overestimated by plate counts . Further experiments must be done to evaluate the lethal efficacy of electrolysis on microorganisms in real conditions encountered in food products.

FEBS Lett, 2003 Sep 25, 552(2-3), 95 - 8
D-Amino acids and D-Tyr-tRNA(Tyr) deacylase: stereospecificity of the translation machine revisited; Yang H et al.; Until 30 years ago, it had been considered that D-amino acids were excluded from living systems except for D-amino acids in the cell wall of microorganisms . However, D-amino acids, in the form of free amino acids, peptides and proteins, were recently detected in various living organisms from bacteria to mammals . The extensive distribution of bio-functional D-amino acids challenges the current concept of protein synthesis: more attention should be paid to the stereospecificity of the translation machine . Besides aminoacyl-tRNA synthetases, elongation factor Tu and some other mechanisms, D-Tyr-tRNA(Tyr) deacylases provide a novel checkpoint since they specifically recycle misaminoacylated D-Tyr-tRNA(Tyr) and some other D-aminoacyl-tRNAs . Their unique structure represents a new class of tRNA-dependent hydrolase . These unexpected findings have far-reaching implications for our understanding of protein synthesis and its origin.

Biotechnol Prog, 2003 Sep-Oct, 19(5), 1410 - 6
Strategic selection of hyperthermophilic esterases for resolution of 2-arylpropionic esters; Sehgal AC et al.; Homologues to Carboxylesterase NP and Candida rugosa lipase, used for the chiral separation of racemic mixtures of 2-arylpropionic methyl esters, were identified by BLAST searches of available genome sequences for hyperthermophilic microorganisms . Two potential candidates were identified: a putative lysophospholipase from Pyrococcus furiosus (Pfu-LPL) and a carboxylesterase from Sulfolobus solfataricus P1 (Sso-EST1) . Although both enzymes showed hydrolytic preference toward the (S) methyl ester, only Sso-EST1 yielded highly optically pure (S) naproxen (%ee(p) >/= 90) and was thus further investigated . Changes in pH or reaction time showed little improvement in %ee(p) or E values with Sso-EST1 . However, the addition of 25% methanol resulted in a 25% increase in E . The effect of various cosolvents on the enantiomeric ratio showed no correlation with the log P or dielectric constant values of the solvent . However, an inverse relationship between E and the denaturation capacity (DC) of the water miscible cosolvents was observed . This was attributed to an increase in enzyme flexibility with increasing solvent DC values leading to a concomitant reduction in the resolving power of Sso-EST1 . The results here show that although bioinformatics tools can be used to select candidate biocatalysts for chiral resolution of 2-arylpropionic esters, biochemical characterization is needed to definitively determine functional characteristics.

Nature, 2003 Oct 2, 425(6957), 521 - 5
Differential regulation of EIN3 stability by glucose and ethylene signalling in plants; Yanagisawa S et al.; Glucose is a global regulator of growth and metabolism that is evolutionarily conserved from unicellular microorganisms to multicellular animals and plants . In photosynthetic plants, glucose shows hormone-like activities and modulates many essential processes, including embryogenesis, germination, seedling development, vegetative growth, reproduction and senescence . Genetic and phenotypic analyses of Arabidopsis mutants with glucose-insensitive (gin) and glucose-oversensitive (glo) phenotypes have identified an unexpected antagonistic interaction between glucose and the plant stress hormone ethylene . The ethylene-insensitive etr1 and ein2 mutants have glo phenotypes, whereas the constitutive ethylene signalling mutant ctr1 is allelic to gin4 (refs 4, 5) . The precise molecular mechanisms underlying the complex signalling network that governs plant growth and development in response to nutrients and plant hormones are mostly unknown . Here we show that glucose enhances the degradation of ETHYLENE-INSENSITIVE3 (EIN3), a key transcriptional regulator in ethylene signalling, through the plant glucose sensor hexokinase . Ethylene, by contrast, enhances the stability of EIN3 . The ein3 mutant has a glo phenotype, and overexpression of EIN3 in transgenic Arabidopsis decreases glucose sensitivity.

Mol Biol Evol, 2004 Jan, 21(1), 97 - 107 Epub 2003 Oct 01.
Sequence determinants of quaternary structure in lumazine synthase; Fornasari MS et al.; Riboflavin, an essential cofactor for all organisms, is biosynthesized in plants, fungi and microorganisms . The penultimate step in the pathway is catalyzed by the enzyme lumazine synthase . One of the most distinctive characteristics of this enzyme is that it is found in different species in two different quaternary structures, pentameric and icosahedral, built from practically the same structural monomeric unit . In fact, the icosahedral structure is best described as a capsid of twelve pentamers . Despite this noticeable difference, the active sites are virtually identical in all structurally studied members . Furthermore, the main regions involved in the catalysis are located at the interface between adjacent subunits in the pentamer . Thus, the two quaternary forms of the enzyme must meet similar structural requirements to achieve their function, but, at the same time, they should differ in the sequence traits responsible for the different quaternary structures observed . Here, we present a combined analysis that includes sequence-structure and evolutionary studies to find the sequence determinants of the different quaternary assemblies of this enzyme . A data set containing 86 sequences of the lumazine synthase family was recovered by sequence similarity searches . Seven of them had resolved three-dimensional structures . A subsequent phylogenetic reconstruction by maximum parsimony (MP) allowed division of the total set into two clusters in accord with their quaternary structure . The comparison between the patterns of three-dimensional contacts derived from the known three-dimensional structures and variation in sequence conservation revealed a significant shift in structural constraints of certain positions . Also, to explore the changes in functional constraints between the two groups, site-specific evolutionary rate shifts were analyzed . We found that the positions involved in icosahedral contacts suffer a larger increase in constraints than the rest . We found eight sequence sites that would be the most important icosahedral sequence determinants . We discuss our results and compare them with previous work . These findings should contribute to refinement of the current structural data, to the design of assays that explore the role of these positions, to the structural characterization of new sequences, and to initiation of a study of the underlying evolutionary mechanisms.

Ophthalmology, 2003 Oct, 110(10), 2051 - 3
Histoplasma granuloma of the conjunctiva; Knox DL et al.; PURPOSE: To describe the clinical and pathologic features of a conjunctival granuloma resulting from Histoplasma capsulatum . DESIGN: Clinicopathologic case report . METHODS: Surgical excision of conjunctival lesion, histopathologic study with special stains, and a fluorescent antibody technique . RESULTS: A conjunctiva granuloma was found to contain microorganisms, compatible with H . capsulatum, confirmed by a fluorescent antibody technique . The conjunctiva returned to normal . CONCLUSIONS: A focal granuloma of the conjunctiva contained H . capsulatum in a patient whose recent symptoms suggested primary infection . The conjunctival lesion was similar to mucocutaneous lesions seen in patients with systemic histoplasmosis.

Biochim Biophys Acta, 2003 Oct 1, 1629(1-3), 44 - 52
Identification of a gene in Leishmania infantum encoding a protein that contains a SP-RING/MIZ zinc finger domain; Garcia-Estrada C et al.; The SP-RING or Miz zinc finger domain that is related to the classical RING-finger motif, defines a class of proteins that can act as E3-like factors in the pathway of small ubiquitin-related modifier (SUMO) conjugation . This family includes the mammalian protein inhibitor of activated STAT (PIAS) proteins and related proteins from lower eukaryotes . Here we report the existence of a gene in Leishmania infantum, present as two identical copies placed upstream of each MAT2 gene copy, and transcribed as a single approximately 2.2 kb mRNA both in the logarithmic and stationary phases of the promastigote stage . This gene encodes a 47 kDa protein that has been named LORIEN . LORIEN is circumscribed to the cell periphery and it is antigenic during L . infantum infection of dogs and hamsters . Strikingly, this novel protein contains a highly conserved SP-RING/Miz zinc finger domain, raising the possibility that a SUMO or ubiquitin-like system may exist in this microorganism.

Tsitologiia, 2003, 45(4), 339 - 56
{Formation and diversity of parasitophorous vacuoles in parasitic protozoa . The Coccidia (Sporozoa, Apicomplexa)}; Beier TV et al.; Data on parasitophorous vacuole (PV) formation in host cells (HC) harbouring different intracellular protozoan parasites have been reviewed and critically analysed, with special reference to the main representatives of the Coccidia . The vacuole membrane (PVM) is the interface between host and parasite, playing a role in nutrient acquisition by the parasite from the HC . The PV phenomenon is regarded as a generalized HC response to the introduction of alien bodies (microorganisms), which eventually reflects the evolutionary established host-parasite relationships at cellular, subcellular and molecular levels . Special attention has been paid to the existing morpho-functional diversity of the PVs within the same genera and species of parasites, and even at different stages of the parasite life cycle . The PVM is generally considered to derive from the HC plasmalemma, whose biochemical composition undergoes significant changes as the intravacuolar parasite grows . The original HC proteins are selectively excluded from the PVM, while those of the parasite are incorporated . As the result, the changed PVM becomes not fusigenic for HC lysosomes . For Toxoplasma gondii and other cyst-forming coccidia (Isospora, Sarcocystis), a definite correlation has been noticed between the extent of rhoptry and dense granule secrets released by a zoite during HC internalization, on the one hand, and the pattern of the PV that forms, on the other one . In T . gondii, tachyzoites, known to discharge abundant secrets, commonly force the development of PVs limited with a single unit membrane and equipped with a tubulovesicular network in the lumen . Unlike, bradyzoites known to be deficient in secretory materials trigger the formation of PVs with a three-membrane lining composed of the changed invaginated plasmalemma in addition to two membranes of endoplasmic reticulum . The two different types of PV harbour, respectively, exoenteric and enteric stages of T . gondii, the latter being confined to the cat intestine only . Unlike, all endogenous stages of the classic intestinal coccidia (Eimeria spp.) develop within PVs limited with a single membrane, with some invaginations extending into the PV lumen . Unusual PV patterns are characteristic of the extracytoplasmic eimerian coccidia (Cryptosporidium, Epieimeria) and adeleid haemogreagarines (Karyolysus) . In cyst-forming coccidia, the PVM is actively involved in tissue cyst wall formation, thus protecting the encysted parasites from recognition by the host immune system . All this strongly suggests that the PV is far from being an indifferent membraneous vesicle containing a parasite, but represents a metabolically active compartment in infected cells . Since all the coccidia are obligate intracellular parasites, the mode of their intimate interaction with the HC, largely accomplished via the PV and its membrane, is vital for their survival as biological species.

Ceska Gynekol, 2003 Jul, 68(4), 254 - 8
{Specific mucosal immunity in the female reproductive tract: protection against infectious diseases and regulation of fertility (part 2)}; Zivny J et al.; Immune mediated defense against pathogenic microorganisms and maintenance of immune tolerance to allogeneic antigens of sperm and fetus are main function of female reproductive tract mucosal immune system . Specific mechanisms concerning development of mucosal vaccines to sexually transmitted diseases and the immune regulation of fertility are reviewed in this article.

J Antibiot (Tokyo), 2003 Jul, 56(7), 639 - 46
Pyrocoll, an antibiotic, antiparasitic and antitumor compound produced by a novel alkaliphilic Streptomyces strain; Dietera A et al.; A new secondary metabolite was detected in the culture extract of Streptomyces sp . AK 409 by HPLC-diode-array screening . The metabolite was identified as pyrocoll, which is known to be a constituent of cigarette smoke . Pyrocoll is known as a synthetic compound, but until now had not been isolated as a natural product from a microorganism . The compound showed biological activity against various Arthrobacter strains, filamentous fungi, several pathogenic protozoa, and some human tumor cell lines.

J Antibiot (Tokyo), 2003 Jul, 56(7), 630 - 8
New haliangicin isomers, potent antifungal metabolites produced by a marine myxobacterium; Kundim BA et al.; Haliangicin is a beta-methoxyacrylate-type polyene antibiotic isolated from the unique marine myxobacterium Haliangium ochraceum . A further investigation of the extract of this microorganism has resulted in the isolation of haliangicin as well as its geometrical isomers . The configuration of the epoxide in haliangicin, which was unknown previously, was determined in this study . The planar structures with partial stereochemistry of these isomers were elucidated by spectroscopic analyses . The major isomer, cis-haliangicin, is a cis isomer of the epoxide portion of haliangicin . Haliangicins B approximately D are geometrical isomers of the polyene moiety, and present as an inseparable mixture of cis and trans epoxide isomers . Their antifungal activities were also evaluated in comparison with haliangicin.

Blood, 2004 Feb 1, 103(3), 836 - 42 Epub 2003 Sep 25.
A potential therapeutic role for small nonpeptidyl compounds that mimic human granulocyte colony-stimulating factor; Kusano K et al.; Granulocyte colony-stimulating factor (G-CSF) stimulates the proliferation of bone marrow granulocytic progenitor cells and promotes their differentiation into granulocytes . G-CSF is therefore an important component of immune defense against pathogenic microorganisms: recombinant human G-CSF (rhG-CSF) is used to treat patients with a variety of neutropenias . In the present study, we screened approximately 10 000 small nonpeptidyl compounds and found 3 small compounds that mimic G-CSF in several in vitro and in vivo assays . These compounds induced G-CSF-dependent proliferation, but had no effect on interleukin-3-dependent, interleukin-2-dependent, interleukin-10-dependent, thrombopoietin (TPO)-dependent, or erythropoietin (EPO)-dependent proliferation . Each compound induced the phosphorylation of signal transducers and activators of transcription-3 (STAT3) and mitogen-activated protein kinase (MAPK) in a G-CSF-dependent cell line and in human neutrophils . In addition, these compounds induced hematopoietic colony formation from primary rat bone marrow cells in vitro . When subcutaneously injected into normal rats, they caused an increase in peripheral blood neutrophil counts . Furthermore, when they were administered to cyclophosphamide-induced neutropenic rats, blood neutrophil levels increased and remained elevated up to day 8 . We therefore suggest that these small nonpeptidyl compounds mimic the activity of G-CSF and may be useful in the treatment of neutropenic patients.

Biochem Biophys Res Commun, 2003 Oct 10, 310(1), 59 - 63
HCV core protein modulates the immune response against the HBV surface antigen in mice; Aguilar JC et al.; Mucosal vaccination is currently arousing a great deal of interest, since mucosally induced immunity is able to protect not only against microorganisms using mucosa as a door of entry, but also against those parenterally transmitted . Hepatitis C virus (HCV) is considered a worldwide health problem and a current vaccine is not available . In the present work, immunogenicity of particulate HCcAg was evaluated, administered alone and also in formulations with the main protective antigen of HBV, the surface antigen (HBsAg), both by mucosal (i.n.) and parenteral (i.m) routes . HCcAg was able to induce strong immune responses after nasal as well as parenteral administration, developing a strong Th1-like antibody response in serum . Preliminary data also suggested the ability of HCcAg to efficiently enhance and modulate the host immune response against HBsAg . These results support the use of the particulate HCcAg in the rational design of candidates for HCV therapeutic or preventive vaccine strategies or inclusively in the development of future combined vaccines.

Eur J Biochem, 2003 Oct, 270(19), 4026 - 38
Purification, characterization and molecular cloning of tyrosinase from the cephalopod mollusk, Illex argentinus; Naraoka T et al.; Tyrosinase (monophenol, L-DOPA:oxygen oxidoreductase) was isolated from the ink of the squid, Illex argentinus . Squid tyrosinase, termed ST94, was found to occur as a covalently linked homodimeric protein with a molecular mass of 140.2 kDa containing two copper atoms per a subunit . The tyrosinase activity of ST94 was enhanced by proteolysis with trypsin to form a protein, termed ST94t, with a molecular mass of 127.6 kDa . The amino acid sequence of the subunit was deduced from N-terminal amino acid sequencing and cDNA cloning, indicating that the subunit of ST94 is synthesized as a premature protein with 625 amino acid residues and an 18-residue signal sequence region is eliminated to form the mature subunit comprised of 607 amino acid residues with a deduced molecular mass of 68,993 Da . ST94 was revealed to contain two putative copper-binding sites per a subunit, that showed sequence similarities with those of hemocyanins from mollusks, tyrosinases from microorganisms and vertebrates and the hypothetical tyrosinase-related protein of Caenorhabditis elegans . The squid tyrosinase was shown to catalyze the oxidation of monophenols as well as o-diphenols and to exhibit temperature-dependency of o-diphenolase activity like a psychrophilic enzyme.

Br J Dermatol, 2003 Sep, 149(3), 627 - 33
Recurrent 'sterile' verrucous cyst abscesses and epidermodysplasia verruciformis-like eruption associated with idiopathic CD4 lymphopenia; Tobin E et al.; Rupture of follicular (epidermoid) cysts is believed to be the consequence of bacterial infection . We report a 24-year-old man with idiopathic CD4 lymphopenia and chronic Mycobacterium avium intracellulare infection who developed multiple, recurring painful abscesses over the distal extremities that increased in number and severity when systemic steroid and interferon-gamma treatment was instituted for interstitial lung disease . Cultures were consistently negative for microorganisms, but pathological examination revealed ruptured epidermoid cyst walls with human papillomavirus (HPV) viropathic changes (keratinocytes with perinuclear halos and abundant basophilic keratohyaline granules) . Cutaneous examination showed numerous, widespread flat-topped papules and achromic macules over the extremities, head and neck . Nested polymerase chain reaction analysis for HPV DNA revealed that the abscess-related cyst walls harboured epidermodysplasia verruciformis (EV)-associated HPV types 20, 24, alb-7 (AY013872) and 80 . His cutaneous lesions harboured HPV types 3, 8 and 80 . Similar to past reports, our patient developed an EV-like eruption in the setting of immunodeficiency . In this instance, EV-associated HPV infection of the follicular infundibular epithelium or pre-existing cysts in the setting of immunodeficiency may have led to cystic growth, rupture and subsequent painful inflammation.

Environ Microbiol, 2003 Oct, 5(10), 925 - 32
Marine prosthecate bacteria involved in the ennoblement of stainless steel; Baker PW et al.; Ennoblement, a phenomenon in which open-circuit potential is elevated to a noble value, triggers metal corrosion in the environment and is considered to be biologically catalysed . This study investigated the involvement of marine microorganisms in the ennoblement of stainless steel coupons in sea water pumped from Kamaishi Bay . Scanning electron microscopy (SEM) showed significant attachment of prosthecate bacteria on the surfaces of stainless steel coupons in the course of ennoblement . In denaturing gradient gel electrophoresis (DGGE) analyses of polymerase chain reaction-amplified bacterial 16S rDNA fragments, several major bands were detected from the surface of the ennobled coupons, including those affiliated with the alpha and gamma subclasses of the Proteobacteria . After these observations, bacterial strains were isolated from the surface of the ennobled coupon . The 16S rDNA analysis revealed that a bacterial isolate (designated PWB3) corresponded to a major DGGE band representing an alpha-Proteobacterial population; a database analysis showed that its closest relative was Rhodobium spp., albeit with low homology ( approximately 89%) . SEM indicated that this bacterium was a prosthecate bacterium that was morphologically similar to those observed on the ennobled coupons . In pure culture of strain PWB3, stainless steel coupons were ennobled when the culture was supplemented with MnCl2 . Manganese was recovered from the surface of the ennobled coupons after treatment with a reducing agent . These results suggest that the attachment of manganese-oxidizing prosthecate bacteria triggered the ennoblement of stainless steel in Kamaishi Bay sea water.

Environ Microbiol, 2003 Oct, 5(10), 878 - 87
Effect of integration of a GFP reporter gene on fitness of Ralstonia eutropha during growth with 2,4-dichlorophenoxyacetic acid; Fuchslin HP et al.; Green fluorescent proteins (GFPs) are frequently used as marker and reporter systems to assess the fate and activity of microbial strains with the ability to degrade xenobiotic compounds . To evaluate the potential of this tool for tracking herbicide-degrading microorganisms in the environment a promoterless gfp was linked to the tfd C promoter, which is activated during degradation of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D), and integrated into the chromosome of the 2,4-D-degrading strain Ralstonia eutropha JMP 134 . The effects of the inserted gfp gene on the kinetics of 2,4-D degradation by R . eutropha in batch and chemostat culture were compared to those of the wild-type strain . In batch culture with 2,4-D as the only carbon and energy source the maximum specific growth rate of the gfp-marked strain did not differ significantly from the wild type . However, compared to the wild type, the 2,4-D steady-state concentration in 2,4-D-limited chemostat cultures of the gfp-marked strain was higher at all dilution rates tested . The reduced competitiveness of the gfp-marked strain at low substrate concentrations was confirmed in a competition experiment for 2,4-D in continuous culture at a dilution rate of 0.075 h-1 . Reproducibly, the gfp-marked strain was displaced by the wild-type strain . The study clearly demonstrates that fitness of constructs cannot be assessed by measuring micro max with selected substrates in batch cultures only but that a thorough kinetic analysis is needed, which also considers slow, carbon-limited growth conditions as they occur in the environment.

Environ Microbiol, 2003 Oct, 5(10), 814 - 27
Stationary phase mutagenesis: mechanisms that accelerate adaptation of microbial populations under environmental stress; Kivisaar M; Microorganisms are exposed to constantly changing environmental conditions . In a growth-restricting environment (e.g . during starvation), mutants arise that are able to take over the population by a process known as stationary phase mutation . Genetic adaptation of a microbial population under environmental stress involves mechanisms that lead to an elevated mutation rate . Under stressful conditions, DNA synthesis may become more erroneous because of the induction of error-prone DNA polymerases, resulting in a situation in which DNA repair systems are unable to cope with increasing amounts of DNA lesions . Transposition may also increase genetic variation . One may ask whether the rate of mutation under stressful conditions is elevated as a result of malfunctioning of systems responsible for accuracy or are there specific mechanisms that regulate the rate of mutations under stress . Evidence for the presence of mutagenic pathways that have probably been evolved to control the mutation rate in a cell will be discussed.

Insect Biochem Mol Biol, 2003 Oct, 33(10), 1025 - 33
Protein purification, cDNA cloning and characterization of a protease inhibitor from the Indian tasar silkworm, Antheraea mylitta; Shrivastava B et al.; An inhibitor of Aspergillus oryzae fungal protease was purified to homogeneity from the hemolymph of fifth instar larvae of Antheraea mylitta by ammonium sulfate precipitation, anion exchange and gel filtration (FPLC) chromatography, and termed as AmFPI-1 . The extent of purification was checked by two-dimensional gel electrophoresis, and the molecular weight of purified inhibitor was determined by SDS-PAGE as 10.4 kDa . Fifteen N-terminal amino acid sequences of this protein were determined, and degenerate oligonucleotides were synthesized on the basis of these sequences . A cDNA library of A . mylitta integument was constructed, and protease inhibitor cDNA was partially amplified by PCR using degenerate oligonucleotides and CDS primers . A full-length inhibitor cDNA clone obtained by screening the library with PCR amplified DNA as probe was sequenced . The cDNA consists of 543 nucleotides with an ORF of 315 bp and encodes a protein of 105 amino acids . The sequence exhibits similarity to several Bombyx mori ESTs, and in particular to N-terminal amino acid sequence of an inducible serine protease inhibitor (ISPI-1) from Galleria mellonella indicating its relatedness to ISPI-1 of G . mellonella . The presence of this protease inhibitor in the hemolymph may play an important role as a natural defense system against invading microorganisms.

Insect Biochem Mol Biol, 2003 Oct, 33(10), 1017 - 24
A serine proteinase homolog venom protein from an endoparasitoid wasp inhibits melanization of the host hemolymph; Asgari S et al.; Activation of prophenoloxidase (proPO) in insects is a defense mechanism against intruding microorganisms and parasites . Pattern recognition molecules induce activation of an enzymatic cascade involving serine proteinases, which leads to the conversion of proPO to active phenoloxidase (PO) . Phenolic compounds produced by pPO-activation are toxic to invaders . Here, we describe the isolation of a venom protein from the parasitoid, Cotesia rubecula, injected into the host, Pieris rapae, which is homologous to serine proteinase homologs (SPH) . The data presented here indicate that the protein interferes with the proteolytic cascade, which under normal circumstances leads to the activation of proPO and melanin formation.

J Food Prot, 2003 Sep, 66(9), 1712 - 5
Inactivation of Saccharomyces cerevisiae with radio frequency electric fields; Geveke DJ et al.; The application of radio frequency (RF) electric fields as a nonthermal alternative to thermal inactivation of microorganisms in liquids was investigated . A novel RF system producing frequencies in the range of 20 to 60 kHz was developed . Electric field strengths of 20 and 30 kV/cm were applied to suspensions of Saccharomyces cerevisiae in water over a temperature range of 35 to 55 degrees C . The flow rate was 1.2 liters/min . The S . cerevisiae population was reduced by 2.1 +/- 0.1 log units following exposure to a 30-kV/cm field at 40 degrees C . The results of the present study provide the first evidence that strong RF electric fields inactivate microorganisms at moderately low temperatures . Increasing the field strength, the number of treatments, and the temperature enhanced inactivation . Frequency had no effect on inactivation over the range of frequencies studied.

J Econ Entomol, 2003 Aug, 96(4), 1045 - 53
Development of a bioassay system for the predator, Xylocoris flavipes (Heteroptera: Anthocoridae), and its use in subchronic toxicity/pathogenicity studies of Beauveria bassiana strain GHA; Dunkel FV et al.; Microbial biocontrol agents are useful commercially only if they do not harm other natural biocontrol organisms, at recommended use rate in the environment where the microorganism is being used . To test the hypothesis that the predatory warehouse pirate bug, Xylocoris flavipes (Reuter), is not adversely affected by the entomopathogen, Beauveria bassiana (Balsamo) Vuillemin strain GHA, we developed a method using individually confined bugs during a 10-d feeding regime . Three concentrations of a conidial suspension were applied to assay surfaces (filter paper disks) to achieve 2.7 x 10(7), 2.6 x 10(6), and 2.6 x 10(5) conidia per cm2, representing 100X, 10X, and a field rate of 2.6 x 10(13) conidia per ha (+/-10%), respectively . Fifth instar X . flavipes suffered 0% infection at the field rate when confined to treated filter paper for 10 d (16% and 42% infection, respectively, at 10X and 100X the field rate) . Second instar migratory grasshoppers, Melanoplus sanguinipes (F.), exposed to the same doses suffered 97, 92, and 100% mortality at the three respective doses 10 d after exposure . These data indicate that B . bassiana can be used safely at recommended application levels without significant effect on fifth instar populations of X . flavipes.

Adv Space Res, 2003, 31(7), 1763 - 8
Survival and alteration of the plasmid-containing microorganism Escherichia coli Z905/pPHL7 introduced into manmade closed aquatic microcosms; Boyandin AN et al.; It has been demonstrated that the transgenic microorganism Escherichia coli Z905/pPHL7 (AprLux+) can exist for a long time at an elevated concentration of mineral salts . The microorganism was introduced into microcosms with sterile brackish water (salinity variable from 21 to 22 g l-1) taken from Lake Shira (Khakasia, Russia) . The survival of the microorganism was estimated both by measuring the growth of the colonies on solid nutrient media and by the bioluminescence exhibited by the transgenic strain in samples from the microcosms and in the enrichment culture with the added selective factor-ampicillin (50 micrograms/ml) . In the enrichment culture, the bioluminescent signal was registered through the 160-day experiment . It has been shown that in the closed microcosms with brackish water the E . coli strain becomes heterogeneous in its ampicillin resistance . The populations of the transgenic strain were mainly represented by isolates able to persist in the medium containing 50 micrograms/ml, but there were also the cells (about 10%) with the threshold of ampicillin resistance not more than 0.05 micrograms/ml . Thus, it was shown that in the microcosms with brackish water and in the absence of the selective factor the transgenic strain survives and retails the recombinant plasmid . c2003 COSPAR . Published by Elsevier Science Ltd . All rights reserved.

Curr Opin Allergy Clin Immunol, 2003 Oct, 3(5), 343 - 6
Innate immunity: sensing the environment and regulating the regulators; Vercelli D; PURPOSE OF REVIEW: Innate immune cells act as sensors for environmental pathogens and key regulators of pathogen-specific T-cell effector responses . The expression of Toll-like receptors is essential for these functions . Innate immunity is a rapidly evolving field . Great progress has been made in the past year in characterizing the signaling pathways downstream of Toll-like receptors, and the role of Toll-like receptors in the regulation of pathogen responses . RECENT FINDINGS: The ability to control the activation of regulatory T cells has emerged as a key function of innate immunity . Regulatory T cells are involved in the induction of tolerance and the prevention of harmful immune pathology, particularly T-cell responses directed against self-antigens . Toll-like receptor-mediated interactions between pathogen-stimulated innate immune cells and regulatory T cells result in the release of suppression by regulatory T cells, thus allowing pathogen-specific responses . However, pathogenic microorganisms may exploit this interaction to evade the host's response . In addition, recent studies raise the possibility that regulatory T cells may express some Toll-like receptor family members, thereby responding directly to pathogens . SUMMARY: A deeper understanding of the complex role of innate immune cells as sensors of the environment and regulators of pathogen responses will probably influence the current models of immune regulation, particularly those centered on the role of the environment in shaping immune responses.

Curr Opin Allergy Clin Immunol, 2003 Oct, 3(5), 337 - 42
Innate immune responses to mycobacteria and the downregulation of atopic responses; Rook GA et al.; PURPOSE OF REVIEW: Exposure to certain environmental microorganisms can promote the induction of T regulatory cells via the innate immune system . This review explores the possibility that reduced exposure to such organisms is leading to increased immunoregulatory disorders in a subset of individuals in whom this regulatory T-cell-inducing pathway is less efficient . We concentrate on mycobacteria and on asthma, because these are well documented . RECENT FINDINGS: The blood cells of the children of farmers, who are partly protected from allergies, express increased levels of messenger RNA encoding CD14 and TLR2, and polymorphisms of CD14 are linked to allergic manifestations in some studies . Polymorphisms of TLR2 (which recognizes mycobacterial components in concert with CD14) are involved in the pattern of response to mycobacteria, and in the type of leprosy that develops . Similarly, polymorphisms of Nramp1, which affect the response to mycobacteria, are linked with the diseases of immunodysregulation that are increasing in parallel with allergic disorders . Moreover, congenic mice bearing different variants of Nramp1 differ in their allergic responses . These parallels are suggestive, in view of the observation that a saprophytic environmental mycobacterium is a potent inducer of regulatory T cells, and has shown significant effects in several phase I/II studies in man . SUMMARY: The components of the innate immune system that are involved in responses to mycobacteria overlap with those implicated in allergic disorders . Polymorphisms might define the subset of individuals who develop immunoregulatory disorders . Understanding the role of the innate immune system will facilitate the design of clinical trials using microbial products.

Proc Natl Acad Sci U S A, 2003 Sep 30, 100(20), 11660 - 5 Epub 2003 Sep 18.
The complete genome sequence of Chromobacterium violaceum reveals remarkable and exploitable bacterial adaptability; Brazilian National Genome Project Consortium; Chromobacterium violaceum is one of millions of species of free-living microorganisms that populate the soil and water in the extant areas of tropical biodiversity around the world . Its complete genome sequence reveals (i) extensive alternative pathways for energy generation, (ii) approximately 500 ORFs for transport-related proteins, (iii) complex and extensive systems for stress adaptation and motility, and (iv) widespread utilization of quorum sensing for control of inducible systems, all of which underpin the versatility and adaptability of the organism . The genome also contains extensive but incomplete arrays of ORFs coding for proteins associated with mammalian pathogenicity, possibly involved in the occasional but often fatal cases of human C . violaceum infection . There is, in addition, a series of previously unknown but important enzymes and secondary metabolites including paraquat-inducible proteins, drug and heavy-metal-resistance proteins, multiple chitinases, and proteins for the detoxification of xenobiotics that may have biotechnological applications.

Infect Immun, 2003 Oct, 71(10), 5733 - 8
The Vsa proteins modulate susceptibility of Mycoplasma pulmonis to complement killing, hemadsorption, and adherence to polystyrene; Simmons WL et al.; The variable surface antigens (Vsa) of the murine respiratory pathogen Mycoplasma pulmonis are associated with the virulence of the microorganism in the lung . In strain UAB CT, the antigens consist of an N-terminal region that is combined with one of seven different C-terminal variable regions comprised of tandem repeats . M . pulmonis producing a VsaA protein with about 40 tandem repeats (R40) does not adhere to red blood cells or polystyrene . Strains that produce VsaH contain a short C-terminal region that lacks tandem repeats and adhere to red blood cells and plastic . We isolated and analyzed M . pulmonis strain CT variants (CT182 and derivatives) that produced a VsaA protein with only three tandem repeats (R3) . These variants adhered to plastic and red blood cells similarly to the VsaH-producing strain . When the R3-producing CT182 strain or the VsaH-producing strains were incubated with normal guinea pig serum, they were efficiently killed . Killing was abolished when the serum was heat inactivated . In contrast, the M . pulmonis strains that produced VsaA R40 were highly resistant to complement killing . CT182R3 variants that survived the complement killing reactions all produced the R40 form of VsaA and were resistant to complement killing . VsaA R40 is the first mycoplasmal protein shown to be associated with resistance to complement . As both VsaH and VsaA can mediate adherence to plastic, cytadherence, and susceptibility to complement, we propose that Vsa modulates these phenotypes by nonspecific interactions.

J Microbiol Methods, 2003 Oct, 55(1), 99 - 103
Spectrofluorimetric method for the estimation of total lipids in Eremothecium ashbyii fungal filaments using Nile blue and avoiding interference of autofluorescent riboflavin; Vijayalakshmi S et al.; A rapid, simple and sensitive spectrofluorimetric technique was developed for monitoring total lipids in hyphae of the riboflavin-overproducing fungus Eremothecium ashbyii using the fluorescent probe Nile blue in an aqueous system, avoiding the interference due to autofluorescent riboflavin . The existing methodologies for lipid estimation are tedious, requiring large biomass, solvent extraction and gravimetry . E . ashbyii is a hemiascomycete fungus which accumulates lipids in its mycelia prior to flavinogenesis . This study defines the conditions (wavelength selection and sensitivity) for the spectrofluorimetric quantification of lipids in situ in the macerated mycelia of this fungus in the presence of intracellular autofluorescent riboflavin without the need to extract the lipids from the mycelia . The fluorescent intensity was linear with the lipid concentration of the mycelia (by gravimetry) under three different growth conditions using glucose, olive oil and sunflower oil as carbon sources . This spectrofluorimetic method of lipid estimation can be applied to other fungi and microorganisms.

J Microbiol Methods, 2003 Oct, 55(1), 65 - 71
Fluorescent in situ hybridization applied on samples taken with adhesive tape strips; La Cono V et al.; Fluorescent in situ hybridization (FISH), applied directly on samples taken with adhesive tape, is proposed as method to detect and identify microorganisms from the surfaces of valuable objects without being destructive . Results of tests carried out in laboratory conditions as well on samples taken from deteriorated surfaces of Roman Catacombs showed the feasibility of FISH when applied on adhesive tape . The potential as well as the limits of the technique were also discussed.

J Microbiol Methods, 2003 Oct, 55(1), 51 - 64
Detection of the biocontrol agent Colletotrichum coccodes (183088) from the target weed velvetleaf and from soil by strain-specific PCR markers; Dauch AL et al.; Diagnostic molecular markers, generated from random amplified polymorphic DNA (RAPD) and used in polymerase chain reaction (PCR), were developed to selectively recognize and detect the presence of a single strain of the biocontrol fungus Colletotrichum coccodes (183088) on the target weed species Abutilon theophrasti and from soil samples . Several isolates of C . coccodes, 15 species of Colletotrichum, a variety of heterogeneous organisms and various plant species were first screened by RAPD-PCR, and a strain specific marker was identified for C . coccodes (183088) . No significant sequence similarity was found between this marker and any other sequences in the databases . The marker was converted into a sequence-characterised amplified region (SCAR), and specific primer sets (N5F/N5R, N5Fi/N5Ri) were designed for use in PCR detection assays . The primer sets N5F/N5R and N5Fi/N5Ri each amplified a single product of 617 and 380 bp, respectively, with DNA isolated from strain 183088 . The specificity of the primers was confirmed by the absence of amplified products with DNA from other C . coccodes isolates, other species representing 15 phylogenetic groups of the genus Colletotrichum and 11 other organisms . The SCAR primers (N5F/N5R) were successfully used to detect strain 183088 from infected velvetleaf plants but not from seeded greenhouse soil substrate or from soil samples originating from deliberate-released field experiments . The sensitivity of the assay was substantially increased 1000-fold when nested primers (N5Fi/N5Ri) were used in a second PCR run . N5Fi/N5Ri selectively detected strain 183088 from seeded greenhouse soils as well as from deliberate-released field soil samples without any cross-amplification with other soil microorganisms . This rapid PCR assay allows an accurate detection of C . coccodes strain 183088 among a background of soil microorganisms and will be useful for monitoring the biocontrol when released into natural field soils.

Biomed Pharmacother, 2003 Sep, 57(7), 269 - 73
Complement and autoimmunity; Boackle SA; The complement system is comprised of a number of serum and membrane-bound proteins that play an important role in the elimination of foreign microorganisms while protecting the host organism from complement-related damage . Complement has also been shown to participate in the generation of normal humoral immune responses to foreign antigens . Recent studies suggest that the functions of complement may be extended to include the maintenance of B cell tolerance . Complement receptor 2 (CR2/CD21) has been implicated in lupus susceptibility in both humans and animal models of disease . Located primarily on B cells and follicular dendritic cells, CR2 binds C3 degradation products that have become covalently bound to antigen or immune complexes in the process of complement activation . The mechanism by which CR2 might regulate B cell reactivity to autoantigens has not been elucidated, but may involve direct effects on B cell tolerance or indirect effects on T cell tolerance.

Biotechnol Adv, 2003 Mar, 21(1), 3 - 39
Biotransformation of drugs by microbial cultures for predicting mammalian drug metabolism; Srisilam K et al.; This review discusses the microbial transformation studies of drugs, correlating them with the corresponding metabolism (biotransformation) in animal systems . Approaches are provided for development of microbial models for mammalian metabolism . Emphasis is placed on the potential of microorganisms to mimic mammalian metabolism and provide ways for structural elucidation and toxicological and pharmacological studies of metabolites . Microorganisms can provide difficult-to-synthesize drugs and assist in identifying metabolic pathways of drugs.

Braz J Infect Dis, 2003 Jun, 7(3), 178 - 93
Leptospirosis in India and the rest of the world; Sambsiava RR et al.; Leptospirosis is an acute anthropo-zoonotic infection of worldwide significance caused by spirochaete Leptospira interrogans which has 23 serogroups and >200 serovars . Various factors influencing the animal activity, suitability of the environment for the survival of the organism and behavorial and occupational habits of human beings can be the determinants of incidence and prevalence of the disease . The disease was considered inconsequential till recently, but it is emerging as an important public health problem during the last decade or so due to sudden upsurge in the number of reported cases and outbreaks . Since isolation rate of the microorganism from clinical specimens is low due to prior indiscriminate use of antibiotics, serological techniques remain the cornerstone of diagnosis.

Int Microbiol, 2003 Dec, 6(4), 233 - 43 Epub 2003 Sep 11.
Innovative tools for detection of plant pathogenic viruses and bacteria; Lopez MM et al.; Detection of harmful viruses and bacteria in plant material, vectors or natural reservoirs is essential to ensure safe and sustainable agriculture . The techniques available have evolved significantly in the last few years to achieve rapid and reliable detection of pathogens, extraction of the target from the sample being important for optimising detection . For viruses, sample preparation has been simplified by imprinting or squashing plant material or insect vectors onto membranes . To improve the sensitivity of techniques for bacterial detection, a prior enrichment step in liquid or solid medium is advised . Serological and molecular techniques are currently the most appropriate when high numbers of samples need to be analysed . Specific monoclonal and/or recombinant antibodies are available for many plant pathogens and have contributed to the specificity of serological detection . Molecular detection can be optimised through the automatic purification of nucleic acids from pathogens by columns or robotics . New variants of PCR, such as simple or multiplex nested PCR in a single closed tube, co-operative-PCR and real-time monitoring of amplicons or quantitative PCR, allow high sensitivity in the detection of one or several pathogens in a single assay . The latest development in the analysis of nucleic acids is micro-array technology, but it requires generic DNA/RNA extraction and pre-amplification methods to increase detection sensitivity . The advances in research that will result from the sequencing of many plant pathogen genomes, especially now in the era of proteomics, represent a new source of information for the future development of sensitive and specific detection techniques for these microorganisms.

Clin Infect Dis, 2003 Oct 1, 37(7), 929 - 32 Epub 2003 Sep 08.
Coronavirus 229E-related pneumonia in immunocompromised patients; Pene F et al.; Coronaviruses strains 229E and OC43 have been associated with various respiratory illnesses ranging from the self-resolving common cold to severe pneumonia . Although chronic underlying conditions are major determinants of severe respiratory virus infections, few data about coronavirus-related pneumonia in immunocompromised patients are available . Here we report 2 well-documented cases of pneumonia related to coronavirus 229E, each with a different clinical presentation . Diagnosis was made on the basis of viral culture and electron microscopy findings that exhibited typical crown-like particles and through amplification of the viral genome by reverse transcriptase-polymerase chain reaction . On the basis of this report, coronaviruses should be considered as potential causative microorganisms of pneumonia in immunocompromised patients.

FEMS Microbiol Lett, 2003 Sep 12, 226(1), 23 - 30
Indole derivatives produced by the fungus Colletotrichum acutatum causing lime anthracnose and postbloom fruit drop of citrus; Chung KR et al.; Postbloom fruit drop (PFD) of citrus and Key lime anthracnose (KLA) are caused by Colletotrichum acutatum . Both fungal isolates can infect flower petals, induce young fruit abscission and result in severe yield loss on many citrus cultivars . Previous studies revealed that infection of citrus flowers by C . acutatum caused higher levels of indole-3-acetic acid (IAA), which could be synthesized from the host plant and/or the fungal pathogen . The ability for IAA production by C . acutatum isolates was investigated . Similar to many microorganisms, the production of indole compounds in the medium by C . acutatum was dependent solely on the presence of tryptophan (Trp) . In total, 14 PFD and KLA fungal isolates were tested, and revealed that they all were capable of utilizing Trp as a precursor to synthesize IAA and other indole derivatives . High-performance liquid chromatography analysis and chromogenic stains after a fluorescence thin-layer chromatography separation unambiguously identified IAA, tryptophol (TOL), indole-acetaldehyde, indole-acetamide (IAM), indole-pyruvic acid, and indole-lactic acid (ILA) from cultures supplemented with Trp . The data suggest that C . acutatum may synthesize IAA using various pathways . Interestingly, increasing Trp concentrations drastically increased the levels of TOL and ILA, but not IAA and IAM . The ability of C . acutatum to produce IAA and related indole compounds may in part contribute to the increased IAA levels in citrus flowers after infection.

Mol Biol Cell, 2003 Sep, 14(9), 3529 - 40 Epub 2003 Jun 27.
The interplay between folding-facilitating mechanisms in Trypanosoma cruzi endoplasmic reticulum; Conte I et al.; Lectin (calreticulin {CRT})-N-glycan-mediated quality control of glycoprotein folding is operative in trypanosomatid protozoa but protein-linked monoglucosylated N-glycans are exclusively formed in these microorganisms by UDP-Glc:glycoprotein glucosyltransferase (GT)-dependent glucosylation . The gene coding for this enzyme in the human pathogen Trypanosoma cruzi was identified and sequenced . Even though several of this parasite glycoproteins have been identified as essential components of differentiation and mammalian cell invasion processes, disruption of both GT-encoding alleles did not affect cell growth rate of epimastigote form parasites and only partially affected differentiation and mammalian cell invasion . The cellular content of one of the already identified T . cruzi glycoprotein virulence factors (cruzipain, a lysosomal proteinase) only showed a partial (5-20%) decrease in GT null mutants in spite of the fact that >90% of all cruzipain molecules interacted with CRT during their folding process in wild-type cells . Although extremely mild cell lysis and immunoprecipitation procedures were used, no CRT-cruzipain interaction was detected in GT null mutants but secretion of the proteinase was nevertheless delayed because of a lengthened interaction with Grp78/BiP probably caused by the detected induction of this chaperone in GT null mutants . This result provides a rationale for the absence of a more drastic consequence of GT absence . It was concluded that T . cruzi endoplasmic reticulum folding machinery presents an exquisite plasticity that allows the parasite to surmount the absence of the glycoprotein-specific folding facilitation mechanism.

J Leukoc Biol, 2004 Jan, 75(1), 27 - 33 Epub 2003 Sep 12.
Collectins and their role in lung immunity; Hickling TP et al.; The collectins are a small family of secreted glycoproteins that contain C-type lectin domains and collagenous regions . They have an important function in innate immunity, recognizing and binding to microorganisms via sugar arrays on the microbial surface . Their function is to enhance adhesion and phagocytosis of microorganisms by agglutination and opsonization . In the lung, two members of the collectin family, surfactant proteins A and D, are major protein constituents of surfactant . Another collectin, mannan-binding lectin, is also present in the upper airways and buccal cavity and may protect against respiratory infections . Recent work has shown that collectins have roles in resistance to allergy and in the control of apoptosis and clearance of apoptotic macrophage in the lung.

Gut, 2003 Oct, 52(10), 1522 - 9
The dendritic cell: its role in intestinal inflammation and relationship with gut bacteria; Stagg AJ et al.; Dendritic cells are antigen presenting cells that are likely to be pivotal in the balance between tolerance and active immunity to commensal microorganisms that is fundamental to inflammatory conditions, including Crohn's disease and ulcerative colitis . Interactions between dendritic cells and microbial products are discussed and how they contribute to regulation of immune responses . The concept that interactions between dendritic cells and commensal organisms may be responsible for maintaining intestinal immune homeostasis is also explored.

Biotechnol Bioeng, 2003 Nov 5, 84(3), 394 - 7
Image analysis of light distribution in a photobioreactor; Jung SK et al.; Light intensity is a crucial factor that determines the growth of photosynthetic cells . This study analyzed the light distribution in a photobioreactor by processing images, captured with a digital camera, of a rectangular photobioreactor containing Synechococcus sp . PCC6801 as a model microorganism . The gray-scale images obtained clearly demonstrate the variation of the light-distribution profiles upon changing cell concentrations and external light intensity . Image-processing techniques were also used to predict the cell density in the photobioreactor . By analyzing the digitized image data with a neural network model, we were able to predict the cell concentrations in the photobioreactor with a <5% error .

Environ Sci Technol, 2003 Sep 1, 37(17), 3885 - 90
Microbial participation in iodine volatilization from soils; Amachi S et al.; The roles of microorganisms in iodine volatilization from soils were studied . Soils were incubated with iodide ion (I-), and volatile organic iodine species were determined with a gas chromatograph . Iodine was emitted mainly as methyl iodide (CH3I), and CH3I emission was sometimes enhanced by the addition of glucose . Soils were then incubated with a radioactive iodine tracer (125I), and radioiodine emitted from soils was determined . The emission of iodine was enhanced in the presence of yeast extract but was inhibited by autoclaving of soils . The addition of streptomycin and tetracycline, antibiotics that inhibit bacterial growth, strongly inhibited iodine emission, while a fungal inhibitor cycloheximide caused little effect . Forty bacterial strains were randomly isolated from soils, and their capacities for volatilizing iodine were determined . Among these, 14 strains volatilized significant amounts of iodine when they were cultivated with iodide ion . Phylogenetic analysis based on 16S ribosomal DNA sequences showed thatthese bacteria are widely distributed through the bacterial domain . Our results suggest that iodine in soils is methylated and volatilized as CH3I by the action of soil bacteria and that iodine-volatilizing bacteria are ubiquitous in soil environments . The pathway of iodine volatilization by soil bacteria should be important for understanding the biogeochemical cycling of iodine as well as for the assessment of long-lived radioactive iodine (129I) in the environment.

Biotechnol Lett, 2003 Jul, 25(14), 1119 - 24
Multiple forms of microbial enzymes; Naessens M et al.; Multiple forms of one enzyme occur in a wide variety of microorganisms . Their synthesis is often dependent on culture characteristics such as medium composition, physico-chemical parameters, culture age and the presence of inducing or inhibiting agents . Multiform enzymes increase the capability of the producing organism to adapt to and cope with a wide variety of environmental changes, such that the physiological advantages outweigh the apparent wasteful hyperproduction of multiple forms of one enzyme.

Zh Mikrobiol Epidemiol Immunobiol, 2003 Jul-Aug, (4), 107 - 9
{Inhibition effect of merthiolate used as vaccines preservative on the oxygen-dependent metabolism of neutrophil granulocytes}; Zinkin VIu et al.; The influence of the newly developed complex vaccine Pyopol, containing the antigens of opportunistic microorganisms and polyoxydonium used as immunomodulator, on the oxygen-dependent metabolism of neutrophils was studied . The study revealed that the main components of the vaccine, both individually and in association, did not change cellular activity in the range of concentrations used in this study . The inhibition of the oxygen-dependent metabolism of neutrophil granulocytes in the presence of native or weakly diluted vaccine occurred due to the cytotoxic effect of thimerosal used as preservative.

Zh Mikrobiol Epidemiol Immunobiol, 2003 Jul-Aug, (4), 104 - 5
{Vitamin K as a stimulator of microbial growth}; Baronets NG; The stimulating action of vitamin K, contained in medicinal plants as vitamin K natural complexes and in Vikasol and Menadion, the pharmaceutical preparations of vitamin K, on the growth of pathogenic microorganisms was determined . The stimulating action of group K vitamins should be taken into consideration in the development of nutrient media.

Zh Mikrobiol Epidemiol Immunobiol, 2003 Jul-Aug, (4), 27 - 31
{Enzyme-substrate mode of action for bacteria survival in water biotic community}; Bukharin OV et al.; The role of enzyme--substrate interactions for the survival of bacteria in natural biocenoses has been analyzed with the systems "lysozyme-antilysozyme", "histon-antihiston" used as models . The conception of a possible universal mechanism for supporting the circulation of pathogenic and opportunistic microorganisms among eukaryotes, irrespective of their evolutionary status and environment, has been formulated . This mechanism is ensured by the natural resistance of the eukaryotic cells and by persistence factors on the part of the pathogen; as a result, the dynamic system of interactions is formed which facilitates the survival of microorganisms due to their persistence potential . New knowledge thus obtained opens prospects in the study of sanitary and ecological aspects of water biocenoses.

Zh Mikrobiol Epidemiol Immunobiol, 2003 Jul-Aug, (4), 8 - 11
{Persistence factors of Helicobacter pylori}; Kirillov VA et al.; Isolates of H . pylori from patients with chronic gastritis and peptic ulcer, were found to be capable of inactivating lysozyme and intercide (the bactericidal component of human leukocytic interferon) . The expression and penetration capacity of their antilysozyme activity (ALA) and antiintercide activity was determined . The wide spread of ALA among H . pylori clinical isolates associated with inflammatory changes in bioptic specimens, confirmed the leading role of this microorganism in the pathogenesis of the gastric mucosal lesions . The retrospective analysis of clinical cases made it possible to recommend the use of the ALA sign as one of the criteria for choosing the scheme of eradication therapy even at the stage of the initial diagnostics of H . pylori infection.

Biotechnol Bioeng, 2003 Oct 20, 84(2), 160 - 9
Titrostat: a reliable technique for continuous cultivation of microorganisms utilizing liquid or gaseous water insoluble substrates; Mutafov S et al.; We describe a method for continuous cultivation of microorganisms utilizing liquid or gaseous water insoluble substrates as a single source of carbon and energy . The water insoluble substrate, which is also the growth-limiting factor, enters the cultivation space as a manually adjustable single-compound material flow . All nonlimiting nutrients (with the exception of oxygen) enter the cultivation space as ingredients of titrant solution which feed rate is reliably coupled to the rate of substrate addition by means of the system titrator.The method provides mild starting conditions appropriate for primary isolation of microorganisms utilizing substrates with growth-inhibiting properties such as BTEX (benzene, toluene, ethylbenzene, and xylene) . The sound control over the microbial specific growth rate makes it suitable for precise kinetic studies as well.We provide a detailed description of both the principles of the method and the equipment used . The dependence of the systems operative range on the concentration of titrant solution is illustrated in the case of continuous cultivation of a mixed bacterial culture on toluene .

Autoimmun Rev, 2003 Sep, 2(5), 258 - 63
An update on Kawasaki disease; Cimaz R et al.; Kawasaki disease (KD) is a febrile systemic vasculitis complicated by coronary and peripheral arterial aneurysms in 20-35% of untreated patients . It is reported as the commonest cause of acquired heart disease in children in developed countries, and may be a risk for adult ischaemic heart disease . Although KD has been reported all over the world, it is overexpressed among Asian populations, especially Japanese . The disease pathogenesis is still unknown and several theories have been proposed, including the possibility of an infection by a toxin-secreting microorganism and of a superantigen-driven process . Despite numerous efforts there is still no diagnostic test available for KD, and the diagnosis is based on clinical criteria after the exclusion of other diseases presenting with high persistent fever . Prompt diagnosis is critical, since the early administration of intravenous immunoglobulins and aspirin reduces the rate of coronary abnormalities to less than 5% of patients.

Braz Dent J, 2003, 14(2), 103 - 8
Healing process of dog teeth after post space preparation and exposition of the filling material to the oral environment; Barbosa HG et al.; The purpose of this study was to observe the influence of coronal leakage on the behavior of periapical tissues after root canal filling and post space preparation . Forty root canals of dogs' teeth were instrumented and filled by the lateral condensation technique with gutta percha points and the cements Sealer 26 and Roth . After post space preparation, the remaining filling material was either protected or not protected with a plug of the temporary cement Lumicon . After root canal exposition to the oral environment for 90 days, the animals were killed and specimens were removed and prepared for histomorphological analysis . The Brown and Brenn technique showed 70% of cases with microorganism leakage for Roth cement, and 20% with Sealer 26 . When a plug of Lumicon was employed, there was 30% leakage for Roth cement and 0% for Sealer 26 . A chronic inflammatory reaction was more frequently observed with Roth cement than with Sealer 26 . It was concluded that a plug of Lumicon was efficient in controlling microorganism coronal leakage (p = 0.05), and that Sealer 26 was more biocompatible and sealed root canals better than Roth sealer (p = 0.01).

IDrugs, 2003 Sep, 6(9), 868 - 73
Microecology as a target for therapeutic intervention in inflammatory bowel disease; Guarner F; The gastrointestinal tract is populated by several hundred grams of bacteria . Recognition of the symbiosis between bacteria and host is drawing particular attention to the implications of bacteria in human health . Probiotics are living microorganisms that upon ingestion in certain numbers exert health benefits . Prebiotics are non-digestible food ingredients that beneficially affect the host by selectively stimulating the growth or activity of a limited number of gut bacteria . In inflammatory bowel disease (IBD), the gut flora is the key factor driving the inflammatory process that leads to intestinal lesions . Interaction of certain commensal bacteria with the gut mucosa triggers mucosal inflammation . However, certain probiotic strains are able to downregulate inflammatory pathways . Restoring the microbial balance with prebiotics and probiotics offers promise for the control of IBD.

Transplant Proc, 2003 Aug, 35(5), 2001 - 3
Visceral leishmaniasis in lung transplantation; Morales P et al.; Infection by intracellular microorganisms with a special geographic distribution, such as Leishmania spp, has been reported in a limited number of patients undergoing solid-organ transplant (SOT) . No cases of Leishmania spp infection in lung transplant patients were found in a review of the literature . In our series of 222 lung or heart and lung transplantations performed from February 1990 to October 2002, two cases of visceral leishmaniasis (VL) were diagnosed and treated with liposomal amphotericin B . All cases reported to date in transplant patients, including the ones discussed here, occurred in people living in or traveling to countries in the Mediterranean area . We therefore consider it advisable to include serological testing for latent infection due to Leishmania spp in pretransplantation screening for our geographical setting, despite the limited return of this strategy.

Nat Biotechnol, 2003 Oct, 21(10), 1229 - 32 Epub 2003 Sep 07.
Electricity generation by direct oxidation of glucose in mediatorless microbial fuel cells; Chaudhuri SK et al.; Abundant energy, stored primarily in the form of carbohydrates, can be found in waste biomass from agricultural, municipal and industrial sources as well as in dedicated energy crops, such as corn and other grains . Potential strategies for deriving useful forms of energy from carbohydrates include production of ethanol and conversion to hydrogen, but these approaches face technical and economic hurdles . An alternative strategy is direct conversion of sugars to electrical power . Existing transition metal-catalyzed fuel cells cannot be used to generate electric power from carbohydrates . Alternatively, biofuel cells in which whole cells or isolated redox enzymes catalyze the oxidation of the sugar have been developed, but their applicability has been limited by several factors, including (i) the need to add electron-shuttling compounds that mediate electron transfer from the cell to the anode, (ii) incomplete oxidation of the sugars and (iii) lack of long-term stability of the fuel cells . Here we report on a novel microorganism, Rhodoferax ferrireducens, that can oxidize glucose to CO(2) and quantitatively transfer electrons to graphite electrodes without the need for an electron-shuttling mediator . Growth is supported by energy derived from the electron transfer process itself and results in stable, long-term power production.

Anesthesiology, 2003 Sep, 99(3), 530 - 5
Hand-cleansing during postanesthesia care; Pittet D et al.; BACKGROUND: Transmission of microorganisms from the hands of healthcare workers is the main source of cross-infection and can be prevented by hand-cleansing . The authors assessed the compliance rate with hand-cleansing practices in the postanesthesia care unit and investigated factors associated with noncompliance . METHODS: Patient care activities, indications for and compliance of postanesthesia care unit staff with hand-cleansing, defined as either washing hands with soap and water or rubbing hands with alcohol, were monitored at the time of patient admission and during their stay . Multivariate analysis identified predictors of noncompliance with hand-cleansing on admission after adjustment for confounders . RESULTS: A total of 3,143 patient care activities, including 1,091 opportunities for hand-cleansing at high or medium risk for cross-transmission, were recorded among 187 patients . The higher the workload, the higher the number of indications for hand-cleansing and the lower the compliance . Average compliance with hand-cleansing at postanesthesia care unit admission was 19.6% . Independent predictors for noncompliance included caring for patients older than 65 yr (odds ratio, 2.23; 95% confidence interval, 1.40-3.57) and those recovering from clean/clean-contaminated surgery (odds ratio, 2.27; 95% confidence interval, 1.11-4.76), as well as high intensity of patient care (odds ratio, 1.01 per patient care activity; 95% confidence interval, 1.0-1.02) . Compliance with hand-cleansing for patients already admitted to the postanesthesia care unit was 12.5% . CONCLUSIONS: Failure to cleanse hands during patient care is common in the postanesthesia care unit and is associated with identifiable factors . The close relation between the intensity of patient care and noncompliance argues that hand-cleansing should not be viewed as a problematic individual behavior only, and system change must be considered in prevention strategies.

Curr Opin Rheumatol, 2003 Sep, 15(5), 528 - 34
Infectious complications in SLE after immunosuppressive therapies; Kang I et al.; Immunosuppressive drugs have become the gold standard for the treatment of major organ involvement in systemic lupus erythematosus . The use of immunosuppressive therapy in systemic lupus erythematosus carries significant risks for infection . This article reviews infectious complications in systemic lupus erythematosus, focusing on effects of immunosuppressive therapy . Patients with systemic lupus erythematosus appear to carry an intrinsically increased risk for infection . Recent studies support this notion further by showing increased risk for serious infections in patients with systemic lupus erythematosus who had mannose-binding lectin deficiency associated with homozygous mannose-binding lectin variant alleles . Patients with systemic lupus erythematosus who were homozygous for mannose-binding lectin variant alleles had a fourfold increase in the incidence of infections, requiring hospitalization . In terms of extrinsic risk factors for infection, use of steroids and cyclophosphamide are the strongest risk factors . The effect of these drugs on infection is also dose dependent . The incidence of infectious complications in patients treated with mycophenolate mofetil, a newly used immunosuppressive drug in systemic lupus erythematosus, appears less frequent compared with cyclophosphamide . Herpes zoster is still the most common viral infection in patients with systemic lupus erythematosus treated with cyclophosphamide and mycophenolate mofetil . Overall data indicate that patients with systemic lupus erythematosus may have intrinsically increased risks for infection that are augmented by immunosuppressive therapies . Cyclophosphamide, in particular in combination with high-dose glucocorticoids, has the strongest effect in suppressing the immune responses against microorganisms . Careful monitoring of infectious complications is warranted in patients with systemic lupus erythematosus receiving immunosuppressive therapies, in particular those on high-dose glucocorticoids and cytotoxic drugs.

J Immunol, 2003 Sep 15, 171(6), 2832 - 9
Sustained response initiation is required for T cell clonal expansion but not for effector or memory development in vivo; Lefrancois L et al.; The factors determining whether an immune response is productive are poorly understood . To understand the circumstances affecting the early stage of the immune response which determine whether memory is generated, the CD8 T cell response was mapped in detail following immunization with live or heat-killed bacteria . Our results demonstrate that even in response to a weak immunogen, functional memory cell development is linked to effector cell induction in lymphoid and nonlymphoid tissues . The main defect in the response to killed microorganisms is inefficient induction of clonal expansion . This failure is due to a contracted, but costimulation-dependent activation phase in the lymphoid tissues, resulting in rapid but abortive growth . Conversely, the response to live bacteria is characterized by protracted early T cell sequestration in lymphoid tissues . Thus, memory development requires effector induction, while optimal clonal expansion is regulated by the duration of response initiation.

Biol Trace Elem Res, 2003 Aug, 94(2), 167 - 77
Effects of La3+ on growth, transformation, and gene expression of Escherichia coli; Wenhua L et al.; Rare earth elements have been emitted into the environment largely as fertilizer components . This has caused much fear about whether they would influence our environment, especially on the metabolism and genetics of microorganisms . In this article, the trivalent ion of a rare earth element, lanthanum, was studied for the effects on growth, transformation, and gene expression of Escherichia coli . The results showed that La3+ at concentrations from 50 to 150 microg/mL stimulated the endogenic metabolism and ectogenic metabolism, but had few effects on gene expression . La3+ at lower concentrations from 0.5 to 30 microg/mL inhibit intensively E . coli-absorbing external DNA, decreasing the transformation efficiency . It is also supported by observations using transmission electron microscopy . Our results are significant in understanding the function of rare earth elements to microorganisms and assessing the risk of application of rare earth compounds.

J Clin Microbiol, 2003 Sep, 41(9), 4068 - 70
Smallpox: residual antibody after vaccination; Gallwitz S et al.; Of all the microorganisms and toxins, poxviruses (Orthopoxvirus) have the greatest potential for use by terrorists . These viruses can spread rapidly through the environment following initial infection . In 1980, the World Health Organization Eradication Program discontinued vaccination for smallpox and declared that the disease had been eliminated . With the threat of smallpox virus as a bioterrorism weapon, questions have been asked about the persistence of protection (as offered by antibodies) following vaccination with vaccinia virus vaccine . To address this, sera from 204 adults vaccinated as children were tested by enzyme immunoassay (EIA) for the presence of vaccinia virus antibody . Of the 204 individuals whose sera were examined for the presence of vaccinia antibody, 165 (80.9%) had been vaccinated once and 39 (19.1%) had been vaccinated at least twice . Of the 165 sera from individuals vaccinated once, 112 (67.9%) were positive . Of the 39 sera from individuals vaccinated more than once, 31 (79.5%) were positive . The presence of a vaccination scar at the time of blood collection was not determined . Fifty-six nonvaccinated individuals, under 30 years of age, were tested by EIA; four of these (7.1%) were positive for vaccinia virus antibody by EIA . Forty-four EIA-positive and 16 EIA-negative sera were also tested by serum neutralization (SN) as a comparison with the EIA test results; one serum (negative by EIA) was SN positive . No attempt was made to ascertain any demographics other than age (date of birth) and "remembered" times of vaccination.

Appl Environ Microbiol, 2003 Sep, 69(9), 5702 - 6
Diversion of electron flow from methanogenesis to crystalline Fe(III) oxide reduction in carbon-limited cultures of wetland sediment microorganisms; Roden EE; Electron flow in acetate-limited cultures of wetland sediment microorganisms was diverted from methane production to Fe(III) reduction in the presence of crystalline Fe(III) oxides at surface area loadings equivalent to that of amorphous Fe(III) oxide . The results indicate that inferences regarding the ability of microbial Fe(III) oxide reduction to compete with other terminal electron-accepting processes in anoxic soils and sediments should be based on estimates of bulk microbially available surface site abundance rather than assumed thermodynamic properties of the dominant oxide phase(s) in the soil or sediment.

Appl Environ Microbiol, 2003 Sep, 69(9), 5519 - 29
Quantitative assessment of picoeukaryotes in the natural environment by using taxon-specific oligonucleotide probes in association with tyramide signal amplification-fluorescence in situ hybridization and flow cytometry; Biegala IC et al.; Picoeukaryotes (cells of <3 micro m in diameter) contribute significantly to marine plankton biomass and productivity, and recently molecular studies have brought to light their wide diversity . Among the methods that have been used so far to quantify aquatic microorganisms, fluorescence in situ hybridization of oligonucleotide probes combined with flow cytometry offers the advantages of both high resolution for taxonomic identification and automated cell counting . However, cell losses, cell clumps, and low signal-to-background ratio have often been mentioned as major problems for routine application of this combination of techniques . We developed a new protocol associating tyramide signal amplification-fluorescence in situ hybridization and flow cytometry, which allows the detection of picoeukaryotes in cultures during both the exponential and stationary phases . The use of surfactant and sonication proved to be essential for the detection and quantification of picoeukaryotes from the natural environment, with as little as a few tenths of a milliliter of 3- micro m-pore-size prefiltered sea water . The routine application of the technique was tested along a coastal transect off Brittany (France), where the different groups of picoeukaryotes were investigated using already published specific probes and a newly designed probe that targets the order Mamiellales (Prasinophyceae, Chlorophyta) . Among the picoeukaryotes, Mamiellales outnumbered by 1 order of magnitude both the cyanobacteria and the non-Chlorophyta, which were represented mainly by the Pelagophyceae class . Picoeukaryote abundance increased from open toward more estuarine water, probably following changes in water temperature and stability.

Biol Chem, 2003 Jul, 384(7), 1035 - 47
Solubilisation and properties of the sialate-4-O-acetyltransferase from guinea pig liver; Iwersen M et al.; The O-acetylation of sialic acids turns out to be one of the most important modifications that influence the diverse biological and pathophysiological properties of glycoconjugates in animals and microorganisms . To understand the functions of this esterification, knowledge of the properties, structures and regulation of expression of the enzymes involved is essential . Attempts to solubilise, purify or clone the gene of one of the sialate-O-acetyltransferases have failed so far . Here we report on the solubilisation of the sialate-4-O-acetyltransferase from guinea pig liver, the first and essential step in the purification and molecular characterisation of this enzyme, by the zwitterionic detergent CHAPS . This enzyme O-acetylates sialic acids at C-4 both free and bound to oligosaccharides, glycoproteins and glycolipids with varying activity, however, gangliosides proved to be the best substrates . Correspondingly, a rapid enzyme test was elaborated using the ganglioside GD3 . The soluble O-acetyltransferase maximally operated at 30 degrees C, pH 5.6, and 50-70 mM KCl and K2HPO4 concentrations . The Km values were 3.6 microM for AcCoA and 1.2 microM for GD3 . CoA inhibits the enzyme with a Ki value of 14.8 microM . A most important discovery enabling further enzyme purification is its need for an unknown low molecular mass and heat-stable cofactor that can be separated from the crude enzyme preparation by 30 kDa ultrafiltration.

Nature, 2003 Sep 4, 425(6953), 75 - 8
Evolution of novel cooperative swarming in the bacterium Myxococcus xanthus; Velicer GJ et al.; Cooperation among individuals is necessary for evolutionary transitions to higher levels of biological organization . In such transitions, groups of individuals at one level (such as single cells) cooperate to form selective units at a higher level (such as multicellular organisms) . Though the evolution of cooperation is difficult to observe directly in higher eukaryotes, microorganisms do offer such an opportunity . Here we report the evolution of novel cooperative behaviour in experimental lineages of the bacterium Myxococcus xanthus . Wild-type strains of M . xanthus exhibit socially dependent swarming across soft surfaces by a mechanism known as 'S-motility' that requires the presence of extracellular type IV pili . In lineages of M . xanthus unable to make pili, a new mechanistic basis for cooperative swarming evolved . Evolved swarming is mediated, at least in part, by enhanced production of an extracellular fibril matrix that binds cells-and their evolutionary interests-together . Though costly to individuals, fibril production greatly enhanced population expansion in groups of interconnected cells . These results show that fundamental transitions to primitive cooperation can readily occur in bacteria.

J Biol Chem, 2003 Nov 21, 278(47), 46337 - 48 Epub 2003 Sep 02.
Evidence of regio-specific glycosylation in human intestinal mucins: presence of an acidic gradient along the intestinal tract; Robbe C et al.; Mucin glycans were isolated from different regions of the normal human intestine (ileum, cecum, transverse and sigmoid colon, and rectum) of two individuals with ALeb blood group . A systematic study of the monosaccharides and oligosaccharide alditols released by reductive beta-elimination from mucins was performed using gas chromatography, matrix-assisted laser desorption ionization time-of-flight mass spectrometry, and nuclear magnetic resonance spectroscopy techniques . Important variations were observed in the mucin-associated oligosaccharide content with an increasing gradient of sialic acid from the ileum to the colon associated with a reverse gradient of fucose . Moreover, a comparative study of the Sda/Cad and ABH blood group determinants along the gastrointestinal tract showed the same reverse distribution in the two kinds of antigens . In addition, besides their heterogeneity, sialic acids presented considerable variations in the degree of O-acetylation in relation to glycan sialylation level . These data are discussed in view of recent concepts suggesting that the oligosaccharide composition of the gut constitutes a varied ecosystem for microorganisms that are susceptible to adapt there and possess the specific adhesion system and specific enzymes able to provide a carbohydrate nutrient.

J Muscle Res Cell Motil, 2002, 23(7-8), 803 - 8
The regulation of phagosome maturation in Dictyostelium; Duhon D et al.; Macropinocytosis (fluid uptake) and phagocytosis (particle uptake) are processes that result in the formation of intracellular membrane enclosed vacuoles termed macropinosomes and phagosomes, respectively . Macropinosomes and phagosomes are modified by fission and fusion reactions with the endo-lysosomal pathway that eventually transform these vacuoles into a lysosomal environment . Many human bacterial pathogens, including species of Mycobacteria, Legionella, and Chlamydia, are thought to survive by disrupting the normal membrane trafficking events that usually result in the formation of phago-lysosomes and death of the microorganism . In addition, a number of important pathogens facilitate homotypic phagosome fusion in order to generate an intracellular environment conducive for survival . A greater understanding of the regulation of phagosomal maturation and fusion will be critical in designing new therapies to treat infections caused by intracellular pathogens . The genetically tractable phagocyte, D . discoideum, has proven extremely useful in dissecting the signaling pathways regulating macropinocytosis, phagocytosis, phagosomal maturation and phagosome-phagosome fusion . A body of knowledge has accumulated and demonstrates important roles for Rab GTPases, the cytoskeleton, phosphoinositide metabolism and pH regulation in regulating phagosome maturation . This review will summarize the current state of knowledge.

Int Endod J, 2003 Sep, 36(9), 604 - 9
Evaluation of time required for recontamination of coronally sealed canals medicated with calcium hydroxide and chlorhexidine; Gomes BP et al.; AIM: To determine in vitro the time required for recontamination of coronally sealed canals medicated with either calcium hydroxide (CaOH2), 2% chlorhexidine gel (CG) or with a combination of both . METHODOLOGY: Eighty intact, caries-free, premolar teeth with straight roots and mature apices were selected for the study . After biomechanical preparation of 75 teeth, they were randomly divided into nine groups according to the intracanal medicament and the coronal seal with 'Intermediate Restorative Material' (IRM) as follows: (i) 10 teeth medicated with CG, coronally unsealed; (ii) 10 teeth medicated with CaOH2, coronally unsealed; (iii) 10 teeth medicated with CaOH2 + CG, coronally unsealed; (iv) 10 teeth medicated with CG + coronal seal; (v) 10 teeth medicated with CaOH2 + coronal seal; (vi) 10 teeth medicated with CG + CaOH2 + coronal seal; (vii) 10 teeth without intracanal medicament and coronally sealed; (viii) 5 teeth without intracanal medicament and coronally unsealed, used as the positive control group (PC); (ix) 5 teeth with intact crowns used as the negative control group (NC) . Glass flasks were filled with Brain Heart Infusion broth (BHI), so that only the root apex was in contact with the broth, while the crown was immersed in human saliva + BHI (3:1) . The flasks were then incubated at 37 degrees C in an atmosphere of 10% CO2, and microbial growth was checked daily . RESULTS: All specimens of the PC showed contamination within 1 day of incubation, while the NC showed no evidence of broth turbidity . Recontamination was detected after an average time of 3.7 days in the unsealed canals medicated with CG, 1.8 days in the group medicated with CaOH2 and 2.6 days in the group medicated with CaOH2 + CG . When the crowns were sealed with IRM, recontamination was detected within 13.5 days in the canals medicated with CG, after 17.2 days in the group medicated with CaOH2 and after 11.9 days in the group medicated with CG + CaOH2 . The group with no medication, but sealed with IRM, showed recontamination after 8.7 days . There were statistically significant differences between the teeth with or without coronal seal (P<0.05) . CONCLUSION: The coronal seal delayed but did not prevent leakage of microorganisms . There was no difference between the various medicaments.

Clin Exp Dermatol, 2003 Sep, 28(5), 531 - 4
Differences between intrafollicular microorganism profiles in perioral and seborrhoeic dermatitis; Takiwaki H et al.; Although often associated with overuse of topical corticosteroids, perioral dermatitis seems to develop seldom in patients with typical seborrhoeic dermatitis in spite of long-term application of corticosteroids . In order to compare the profiles of intrafollicular microorganisms in the lesions of perioral and seborrhoeic dermatitis, tape-stripped samples were obtained from eight lesions of perioral dermatitis, 10 lesions of seborrhoeic dermatitis, and the perioral skin of 31 normal subjects . After staining with Toluidine blue, resident microorganisms on plucked hair roots were evaluated microscopically . In all patients with perioral dermatitis and two normal subjects, 20-70% of sample hairs were positive for fusiform bacteria regarded as fusobacteria . Malassezia-positive hairs were rarely seen in these cases . Seborrhoeic dermatitis showed the opposite results . Perioral dermatitis may tend to develop under fusiform-bacteria-rich conditions, rather than Malassezia-rich conditions as in the case of seborrhoeic dermatitis.

J Gen Appl Microbiol, 2003 Jun, 49(3), 141 - 54
Isolation of new actinomycete strains for the screening of new bioactive compounds; Takahashi Y et al.; In order to facilitate the discovery of novel bioactive compounds from microorganisms, various techniques for isolation of new actinomycete strains have been attempted . Studies of the vertical distribution of actinomycetes in soil, isolation of actinomycetes from desert soils or fallen leaves, selective isolation of Kitasatospora strains using novobiocin or Actinoplanes strains using the chemotactic method, and the use of gellan gum as a solidifying agent were carried out . We discovered 9 novel bioactive compounds from actinomycete strains isolated under unusual conditions, and proposed two new genera, five new species and one new subspecies.

Biochim Biophys Acta, 2003 Sep 2, 1615(1-2), 60 - 8
Ca(2+) sequestering in the early-branching amitochondriate protozoan Tritrichomonas foetus: an important role of the Golgi complex and its Ca(2+)-ATPase; Almeida JC et al.; Total membrane vesicles isolated from Tritrichomonas foetus showed an ATP-dependent Ca(2+) uptake, which was not sensitive to 10 microM protonophore FCCP but was blocked by orthovanadate, the inhibitor of P-type ATPases (I(50)=130 microM), and by the Ca(2+)/H(+) exchanger, A-23187 . The Ca(2+) uptake was prevented also by thapsigargin, an inhibitor of the SERCA Ca(2+)-ATPases . The sensitivity of the Ca(2+) uptake by the protozoan membrane vesicles to thapsigargin was similar to that of Ca(2+)-ATPase from rabbit muscle sarcoplasmic reticulum . Fractionation of the total membrane vesicles in sucrose density gradient revealed a considerable peak of Ca(2+) transport activity that co-migrated with the Golgi marker guanosine diphosphatase (GDPase) . Electron microscopy confirmed that membrane fractions of the peak were enriched with the Golgi membranes . The Golgi Ca(2+)-ATPase contributed to the Ca(2+) uptake by all membrane vesicles 80-85% . We conclude that: (i) the Golgi and/or Golgi-like vesicles form the main Ca(2+) store compartment in T . foetus; (ii) Ca(2+) ATPase is responsible for the Ca(2+) sequestering in this protozoan, while Ca(2+)/H(+) antiporter is not involved in the process; (iii) the Golgi pump of this ancient eukaryotic microorganism appears to be similar to the enzymes of the SERCA family by its sensitivity to thapsigargin.

J Environ Monit, 2003 Aug, 5(4), 667 - 70
Restoration of properties of cultivated soils polluted by copper and nickel; Evdokimova GA et al.; The long-term observations of the restoration of chemical and biological properties of cultivated podzol soil polluted by airborne emissions from a copper-nickel smelter located in the western Arctic of Russia were carried out . After 8 years, the total content of copper in soil decreased to a third (from 6230 to 2080 mg kg(-1)) and nickel to a sixth (from 3500 to 580 mg kg(-1)) . Based on these observations, the removal time for heavy metals to reach permissible levels was calculated . The estimate was 100 years for copper and 108 years for nickel . Soil remained toxic to wheat seedlings, especially to their roots, for the whole observation period . A restoration of the number and activity of soil microorganisms was associated with the reduction in soil metal-toxicity . The number of non-sporeforming saprophytic bacteria was quickly restored concurrently with the removal of metals from the soil; fungal biomass also increased.

Biodegradation, 2003 Aug, 14(4), 285 - 95
H2 consumption during the microbial reductive dehalogenation of chlorinated phenols and tetrachloroethene; Mazur CS et al.; Competition for molecular hydrogen exists among hydrogen-utilizing microorganisms in anoxic environments, and evidence suggests that lower hydrogen concentrations are observed with more energetically favorable electron-accepting processes . The transfer of electrons to organochlorines via reductive dehalogenation reactions plays an important role in hydrogen dynamics in impacted systems . We studied the flux of aqueous hydrogen concentrations in methanogenic sediment microcosms prior to and during reductive dehalogenation of a variety of substituted chlorophenols (CP) and tetrachloroethene (perchloroethylene, PCE) . Mean hydrogen concentrations during reductive dehalogenation of 2,4-CP, 2,3,4-CP, and PCP were 3.6 nM, 4.1 nM, and 0.34 nM, respectively . Sediment microcosms that were not dosed with chlorophenols yet were actively methanogenic maintained a significantly higher mean hydrogen concentration of 9.8 nM . During active PCE dehalogenation, sediment microcosms maintained a mean hydrogen concentration of 0.82 nM . These data indicate that during limiting hydrogen production, the threshold ecosystem hydrogen concentration is controlled by microbial populations that couple hydrogen oxidation to thermodynamically favorable electron accepting reactions, including reductive dehalogenation of chloroaromatic and chloroaliphatic compounds . We also present revised estimates for the Gibbs free energy available from the reductive dehalogenation of a variety of substituted chlorophenols based on recently published values of vapor pressure, solubility, and pKa for these compounds.

Mol Biochem Parasitol, 2003 Aug 31, 130(2), 65 - 74
Leishmania lipophosphoglycan (LPG) activates NK cells through toll-like receptor-2; Becker I et al.; Toll-like receptors (TLRs) mediate the cellular response to conserved molecular patterns shared by microorganisms . We report that TLR-2 on human NK cells is upregulated and stimulated by Leishmania major lipophosphoglycan (LPG), a phosphoglycan belonging to a family of unique Leishmania glycoconjugates . We found that purified L . major LPG upregulates both mRNA and the membrane expression of TLR-2 in NK cells . Additionally, IFN-gamma and TNF-alpha production and nuclear translocation of NF-kappaB was enhanced . The activation effect was more intense with LPG purified from infectious metacyclic parasites than from noninfectious procyclic Leishmania . Since the difference between the molecules derived from these two stages of the parasite growth cycle lies exclusively in the number of phosphosaccharide repeat domains and in the composition of glycan side chains that branch off these domains, we propose that TLR-2 possibly distinguishes between phosphorylated glycan repeats on LPG molecules . The effect of LPG on cytokine production and on membrane expression of TLR-2 could be blocked with F(ab')2 fragments of the mAb against LPG (WIC 79.3) . Confocal microscopy demonstrated the co-localization of LPG and TLR-2 on the NK cell membrane . Binding of LPG to TLR-2 in NK cells was demonstrated by immunoprecipitations done with anti-TLR-2 and anti-LPG mAb followed by immunoblotting with anti-LPG and anti-TLR-2, respectively . Both antibodies recognized the immune complexes . These results suggest that NK cells are capable of recognition of, and activation by, Leishmania LPG through TLR-2, enabling them to participate autonomously in the innate immune system and thereby increasing the effective destruction of the parasite.

Phytochemistry, 2003 Sep, 64(1), 199 - 207
Early steps of deoxyxylulose phosphate pathway in chromoplasts of higher plants; Fellermeier M et al.; 1-Deoxy-D-xylulose 5-phosphate and 2C-methyl-D-erythritol 4-phosphate have been shown as intermediates of the deoxyxylulose phosphate pathway used for terpenoid biosynthesis in plants and many microorganisms . In plants this non-mevalonate pathway is located in plastids . In order to investigate the formation of five carbon intermediates, chromoplasts from Capsicum annuum and Narcissus pseudonarcissus were incubated with isotope-labeled 1-deoxy-D-xylulose 5-phosphate or 2C-methyl-D-erythritol 4-phosphate . The downstream metabolites were detected and separated by reversed-phase ion-pair radio-HPLC and their structures elucidated by mass spectroscopy . Here we report the isolation and structural identification of 4-diphosphocytidyl-2C-methyl-D-erythritol and 2C-methyl-D-erythritol 2,4-cyclodiphosphate from chromoplasts; the genes of the corresponding enzymes had been previously identified from Escherichia coli and Arabidopsis.

Eur Respir J Suppl, 2003 Aug, 42, 72s - 76s
Prevention of nosocomial infections in acute respiratory failure patients; Girou E; Patients with acute respiratory failure are predisposed to acquire nosocomial infection primarily because they may need ventilatory support, usually invasive mechanical ventilation . The presence of an endotracheal tube impairs natural defences of the respiratory tract and favours airways colonisation and lung infection . Cross transmission of microorganisms may also occur via contaminated hands of healthcare workers that manipulate invasive devices . Thus, avoiding the endotracheal tube and increasing hand hygiene compliance are major measures to prevent ventilator-associated pneumonia . The use of noninvasive ventilation has been shown to reduce the incidence of nosocomial infections and should be used whenever possible . Using hand rubbing with waterless alcohol-based products evidenced higher efficacy to reduce hand contamination as compared to conventional hand washing with soap . Due to its rapid activity and ease of access, hand rubbing constitutes a promising avenue for improving hand hygiene compliance and thus reducing cross infection.

Eur Respir J Suppl, 2003 Aug, 42, 10s - 14s
Inflammatory response to infectious pulmonary injury; Delclaux C et al.; This review describing the inflammatory response to infectious pulmonary injury is focused on the innate immunity of the distal lung to bacterial pneumonia . The fact that the inflammatory response varies to some extent with the bacterial strain responsible for the infection is emphasised . The key cellular components present in the distal lung are described . The major role of alveolar macrophage is described, inasmuch as it responds to the usual daily challenges of bacteria entering the terminal airways and is capable of initiating an inflammatory reaction if the microbial challenge is either too large or too virulent . Under these conditions, the alveolar macrophages initiate an inflammatory response that recruits large numbers of neutrophils into the alveolar spaces . The strategy of the innate immune response may not be to recognise every possible antigen, but rather to focus on a few, highly conserved structures present in large groups of microorganisms . These structures are referred to as pathogen-associated molecular patterns and the receptors of the innate immune system that evolved to recognise them are called pattern-recognition receptors . The soluble factors in innate defence, such as cytokines, are described, and a last paragraph discusses whether a specific inflammatory response could characterise nosocomial pneumonia.

Arch Latinoam Nutr, 2003 Mar, 53(1), 65 - 9
{Effect of microwave oven over the growth and survival of Escherichia coli O157:H7 inoculated in bovine minced meat samples}; Quesada O et al.; The use of microwave ovens in food industry is a growing trend . It is used for thawing, drying and cooking food, but the microorganism's inactivation that this treatment may exert or not is still a subject of worldwide discussion . At the same time . Escherichia coli O157:H7 now presents itself as an emerging pathogen, distributed worldwide and associated with food . Its resistance to adverse environments has been widely discussed . The purpose of this work was to determine the effect of different times of exposure and cooking intensities of microwave oven on the survival of this bacterium inoculated into minced meat samples . These were inoculated with a high (10(7)-10(9) CFU/mL) or low (10(7)-10(7) CFU/mL) population of E . coli O157:H7, frozen for 3 days at-4 degrees C and thawed in a Whirlpool microwave oven according to their weight . They were radiated at levels of 70%, 80%, 90% and 100% for periods of 30, 60, 90 and 120 seconds . In each sample the rate of survival of the bacteria was determined according to the methodology described by Vanderzant & Splittstoesser . The results obtained showed that the rate of destruction of the bacteria analyzed was significant (p < 0.005) . The number of bacteria present in the meat samples diminished dramatically as the exposure time and temperature increased, even though, for the complete elimination of this microorganism, a prolonged exposure was necessary, even though it did cause undesirable organoleptic characteristics in the food samples.

Mycorrhiza, 2004 Aug, 14(4), 221 - 8 Epub 2003 Aug 29.
Arbuscular mycorrhizal fungi colonize decomposing leaves of Myrica parvifolia, M . pubescens and Paepalanthus sp; Aristizabal C et al.; Hyphae and vesicles of arbuscular mycorrhizal fungi (AMF) were found within the decomposing leaves of Myrica parvifolia, M . pubescens and Paepalanthus sp . at three montane sites in Colombia . Hyphae, vesicles, and arbuscule-like structures were also found within scale-like leaves of the rhizomes of Paepalanthus sp . The litter found in the vicinity of the roots was divided into three decomposition layers . The highest AMF colonization occurred in the most decomposed leaves, which were in close association with roots . In contrast, there were no differences in AMF colonization of roots present in the different decomposition layers . Colonization of decomposing leaves by AMF did not differ between the two closely related species M . parvifolia and M . pubescens, nor between two sites (Guatavita and Zipacon, Colombia) differing in soil fertility . Occurrence of vesicles in decomposing leaves was correlated with abundant AMF extraradical hyphae among the leaves . We propose that AMF enter decomposing leaves mechanically through vascular tissue . As a consequence, AMF are well positioned to obtain and efficiently recycle mineral nutrients released by decomposer microorganisms before their loss by leaching or immobilization in soil.

J Prosthet Dent, 2003 Sep, 90(3), 282 - 8
Calcium hypochlorite as a disinfecting additive for dental stone; Twomey JO et al.; STATEMENT OF PROBLEM: Dental casts come into direct contact with impression materials and other items that are contaminated by saliva and blood from a patient's mouth, leaving the casts susceptible to cross-contamination . Topical methods of disinfecting casts are difficult to control, while immersion methods are potentially destructive . Thus, an additional method to control cross-contamination between patients and laboratory personnel is needed . PURPOSE: This study was undertaken in an attempt to develop a dental stone with disinfecting properties and adequate compressive and tensile strengths . MATERIAL AND METHODS: Calcium hypochlorite {Ca(OCl)(2)} in aqueous solution in concentrations from 0 to 1.5% was tested as a disinfecting additive to type V dental stone . The compressive and tensile strength properties of the modified stone were measured (MPa) using a universal testing machine at a consistency similar to unmodified stone . Strength data were analyzed by 1-way ANOVA and post hoc Tukey-Kramer procedure (alpha < or =.05) . To measure the disinfecting ability, the effect on Bacillis subtilis bacteriophage phi29 was tested in triplicate to find the minimum concentration at which no phage was detected . Additionally, 3 impressions were disinfected with CaviCide, and 3 impressions rinsed in water served as controls . RESULTS: In general, the effect of adding the disinfectant to the stone was a decrease in strength . Exceptions were the dry compressive strength, for which there was a significant increase in strength (P=.048) at 0.5%, and the wet compressive and wet tensile strength, which showed no significant difference between the 1.5% and the control . When Ca(OCl)(2) was added at the concentration 0.5% (2765 ppm available chlorine), the gypsum had acceptable mechanical properties; dry compressive strength was 78.86 +/- 4.12 MPa, and dry tensile strength was 10.64 +/- 1.27 MPa, compared to control values of 67.85 +/- 6.28 and 13.41 +/- 1.24 MPa, respectively . At concentrations of 0.3% and higher (36 1650 ppm of available chlorine), calcium hypochlorite was able to completely inactivate phi29 . CONCLUSION: It is possible to prepare a type V dental stone that contains a disinfectant, has adequate mechanical properties, and will reduce numbers of residual microorganisms . For example, stone mixed with water containing 0.5% Ca(OCl)(2) meets these criteria.

Microbes Infect, 2003 Sep, 5(11), 1013 - 21
Infections with the chlamydia-like microorganism Simkania negevensis, a possible emerging pathogen; Friedman MG et al.; Although evidence for the existence of numerous chlamydia-like microorganisms has been discovered in both environmental samples and clinical specimens, very few have been grown in vitro, and little is known of their pathogenic potential . Of all such organisms, Simkania negevensis is probably the most extensively studied . This review summarizes current knowledge about this intracellular bacterium, focusing especially on human infections.

Proc Natl Acad Sci U S A, 2003 Sep 16, 100(19), 10983 - 8 Epub 2003 Aug 25.
Genetic identification of a respiratory arsenate reductase; Saltikov CW et al.; For more than a decade, it has been recognized that arsenate {H2AsO41-; As(V)} can be used by microorganisms as a terminal electron acceptor in anaerobic respiration . Given the toxicity of arsenic, the mechanistic basis of this process is intriguing, as is its evolutionary origin . Here we show that a two-gene cluster (arrAB; arsenate respiratory reduction) in the bacterium Shewanella sp . strain ANA-3 specifically confers respiratory As(V) reductase activity . Mutants with in-frame deletions of either arrA or arrB are incapable of growing on As(V), yet both are able to grow on a wide variety of other electron acceptors as efficiently as the wild-type . Complementation by the wild-type sequence rescues the mutants' ability to respire As(V) . arrA is predicted to encode a 95.2-kDa protein with sequence motifs similar to the molybdenum containing enzymes of the dimethyl sulfoxide reductase family . arrB is predicted to encode a 25.7-kDa iron-sulfur protein . arrA and arrB comprise an operon that contains a twin arginine translocation (Tat) motif in ArrA (but not in ArrB) as well as a putative anaerobic transcription factor binding site upstream of arrA, suggesting that the respiratory As(V) reductase is exported to the periplasm via the Tat pathway and under anaerobic transcriptional control . These genes appear to define a new class of reductases that are specific for respiratory As(V) reduction.

Occup Environ Med . 2003 Sep;60(9):E5.
Inflammatory potential of dust from schools and building related symptoms; Allermann L et al.; AIMS: To test whether the inflammatory potential of dust samples might be used to differentiate schools with high and low prevalence of building related symptoms (BRS) among the occupants . METHODS: Ten schools with high prevalence of BRS and 10 schools with low prevalence were selected . Dust collected from floors, horizontal surfaces, and exhaust outlets was tested at five concentrations on the lung epithelial cell line A549 . The potency of the dust (PF) to stimulate IL-8 secretion was calculated from the initial linear part of the dose-response curves . The organic fraction of the dust samples was determined by incineration . RESULTS: The schools with low prevalence of symptoms had a BRS% of 4.4-11.0 and the schools with high prevalence a BRS% of 19.6-31.9 . The PF of floor dust and surface dust correlated, and the PF was associated with the organic content of the dust . The schools with low prevalence of symptoms had a significantly lower PF than the schools with high prevalence . Using the cut point value of 4.5 ng IL-8/mg floor dust, significantly more high prevalence schools were found above the cut point than below . CONCLUSION: The PF of the floor dust samples correlated significantly with the prevalence of symptoms in the schools . The content of endotoxin and microorganisms did not seem to explain the inflammatory potential of the dust or BRS, and the substances in the dust causing the inflammatory potential are presently unknown.

Leg Med (Tokyo), 2001 Mar, 3(1), 15 - 22
Postmortem bacteriology in forensic pathology: diagnostic value and interpretation; Tsokos M et al.; The proof of a pathogenetic germ can be of evidential value for the medico-legal expertise concerning etiopathogenetic conclusions towards a causal relationship between infection and fatal outcome . In specific cases the forensic pathologist may be confronted with the decision whether cultures for postmortem bacteriological investigations should be obtained at autopsy . The present review surveys the value and practicability of postmortem bacteriological cultures in the light of the literature . The most promising media for postmortem bacteriological cultures are spleen and heart blood . In contrast, postmortem lung culture results are often unreliable due to frequently false positive results . The collection of specimens from at least two different sampling sites should be the standard procedure in cases where an underlying (potentially fatal) infection is presumed . Multiple postmortem cultures from different sampling sites raise the probability to identify the etiologic agent of antemortem infection . A careful correlation of culture results from different anatomical sampling sites with autopsy and histological findings is essential for the differentiation between vitally acquired infection and postmortem contamination . Various factors (e.g . the time interval between death and sampling of specimens, agonal spread of microorganisms or preceding antibiotic therapy prior to death) may limit the diagnostic utility of postmortem culture results . Contamination of specimens may, e.g . originate from inadequate sampling techniques, use of unsuitable transport media or postmortem growth of bacteria . The number of contaminated postmortem cultures increases when bowel manipulation or evisceration occurs prior to sampling and is depending on the anatomy of the sampling site . Polymicrobial growth can be considered contamination in the majority of cases . For the medico-legal expertise a thorough histological examination of inner organs and tissues is needed; the histological section has to represent the sampling site for culture . The presence of an underlying infection can be proven when postmortem bacteriological cultures yield a primary infectious pathogen and a cellular response in the affected tissue can be detected on the histopathological level . Postmortem bacteriology can be regarded as a reliable diagnostic tool for postmortem quality control of preceding diagnostic and therapeutic procedures and can serve as an additional indicator of nosocomial infections within a specific hospital environment . From the medico-legal point of view, a more widespread application of this valuable investigative procedure, that adds to diminish the number of otherwise unresolved deaths in forensic autopsy practice, is desirable.

Klin Lab Diagn, 2003 Jul, (7), 48 - 50
{Use of nested PCR in detection of the plague pathogen}; Glukhov AI et al.; Causative agents of plague, i.e . bacterium Yersina pestis (in the subcutaneous tissues of rodents) and their cutaneous parasites need to be isolated to enable plague prevention . A comparatively new method of polymerase chain reaction (PCR) opens up new possibilities of determining Y . pestis just within several hours and without any cultivation . The article contains a description of the PCR-method, which makes it possible to distinguish the culture of Y . pestis from cultures of other microorganism, including speci of Yersina . The method is of the cluster-type, i.e . it is made up of subsequent PC reactions with the substrate for the second reaction being the product of the first one . The cluster nature of the method preconditions a higher sensitivity and specificity versus the ordinary PCR.

J Infect Dis, 2003 Sep 1, 188(5), 777 - 82 Epub 2003 Aug 08.
Mannose-binding lectin polymorphisms in clinical tuberculosis; Soborg C et al.; Mannose-binding lectin (MBL) mediates protection against infections by using the complement system, but certain microorganisms may increase infectivity by exploiting this host defense system . Thus, it has been speculated whether genetically determined low MBL levels may confer partial protection against certain intracellular microorganisms, such as Mycobacterium tuberculosis . We investigated MBL alleles in 109 culture-positive human immunodeficiency virus-uninfected patients with tuberculosis living in Denmark and 250 white control subjects . Patients and control subjects were divided into 3 different groups defined by undetectable, low, and high serum MBL concentrations, which correlates to deficient, low, and high expressing MBL genotypes . A significantly decreased frequency of patients with the low-expressing MBL genotype was observed in white patients compared to control subjects . The same tendency also was observed in patients of other ethnic origin . It may be hypothesized that heterozygosity for MBL variant alleles, which encodes low serum MBL levels, is associated with protection against clinical tuberculosis.

Infect Immun, 2003 Sep, 71(9), 5254 - 65
Protective mucosal Th2 immune response against Toxoplasma gondii by murine mesenteric lymph node dendritic cells; Dimier-Poisson I et al.; Toxoplasma gondii, an obligate intracellular parasite pathogen which initially invades the intestinal epithelium before disseminating throughout the body, may cause severe sequelae in fetuses and life-threatening neuropathy in immunocompromised patients . Immune protection is usually thought to be performed through a systemic Th1 response; considering the route of parasite entry it is important to study and characterize the local mucosal immune response to T . gondii . Despite considerable effort, Toxoplasma-targeted vaccines have proven to be elusive using conventional strategies . We report the use of mesenteric lymph node dendritic cells (MLNDCs) pulsed ex vivo with T . gondii antigens (TAg) as a novel investigation approach to vaccination against T . gondii-driven pathogenic processes . Using a murine model, we demonstrate in two genetically distinct mouse strains (C57BL/6 and CBA/J) that adoptively transferred TAg-pulsed MLNDCs elicit a mucosal Toxoplasma-specific Th2-biased immune response in vivo and confer strong protection against infection . We also observe that MLNDCs mostly traffic to the intestine where they enhance resistance by reduction in the mortality and in the number of brain cysts . Thus, ex vivo TAg-pulsed MLNDCs represent a powerful tool for the study of protective immunity to T . gondii, delivered through its natural route of entry . These findings might impact the design of vaccine strategies against other invasive microorganisms known to be delivered through digestive tract.

Int J Phytoremediation, 2003, 5(2), 105 - 23
Ability of cold-tolerant plants to grow in hydrocarbon-contaminated soil; Robson DB et al.; Phytoremediation of hydrocarbons in soil involves plants and their associated microorganisms . Differences in environmental conditions and restrictions on species importation mean that each country may need to identify indigenous plants to use for phytoremedation . Screening plants for hydrocarbon tolerance before screening for degradation ability may prove more economical than screening directly for degradation . Thirty-nine cold-tolerant plants native, or exotic and naturalized, in western Canada were assessed for their ability to survive in crude oil-contaminated soil . Four naturalized grasses (i.e., Agropyron pectiniforme, Bromus inermis, Phleum pratense, and Poa pratensis), three naturalized legumes (i.e., Medicago sativa, Melilotus officinalis, and Trifolium repens), two native forbs (i.e., Artemisia frigida and Potentilla pensylvanica), one native grass (i.e., Bromus ciliatus) and two native legumes (i.e., Glycyrrhiza lepidota and Psoralea esculenta) exhibited phytoremediation potential, based on survival . We determined the effect of increasing crude oil concentrations on total and root biomass, and relative growth rate of those species with the highest survival . The addition of 0.5%, 1%, and 5% (crude oil wt/fresh soil wt) crude oil to soil significantly decreased both the total biomass by at least 22% of the control and the relative growth rate of all species except P . esculenta . Root biomass significantly decreased by at least 22% with crude oil addition in all species except P . esculenta and A . frigida . Total biomass production in contaminated soil had a significant negative correlation with the relative growth rate in uncontaminated soil.

Ecotoxicol Environ Saf, 2003 Oct, 56(2), 265 - 70
Vermicomposting of industrially produced woodchips and sewage sludge utilizing Eisenia fetida; Maboeta MS et al.; Adult Eisenia fetida were used to vermicompost woodchips (WC) and sewage sludge (SS) that are produced as waste product by platinum mines . The aims of the study were to examine the growth and reproductive success of the worms over 84 days to determine long-term feasibility of large-scale implementation and monitor the bioconcentration of heavy metals and the effects of microorganisms inoculation to quantify possible environmental implications . Results revealed that there were no effects on growth (P>0.05), reproductive success decreased (P<0.05), and aluminum (Al), copper (Cu), and nickel (Ni) were bioconcentrated (P<0.05) in the treatment groups without an inoculate . Earthworms in the treatment group with the microorganism inoculate manifested no effects on growth or reproductive success and did not accumulate Al, Cu, and Ni . It is concluded that the only economically feasible way to bioconvert WC and SS to a potential ameliorant of platinum mine tailings would be with the addition of a microorganism inoculate.

Forensic Sci Int, 2003 Aug 27, 135(3), 226 - 31
Investigating demodex in forensic autopsy cases; Ozdemir MH et al.; Demodex is an ectoparasite living in the skin as a nonpathogen or a pathogen . It is also known that demodex acts as a vector of pathogenic microorganisms . In this study, we have investigated the rate of occurrence and vitality of demodex in forensic autopsies that have a high risk of contamination by infected organisms . The study, which was cross-sectional, conducted on 100 autopsy cases: 77 of the bodies were male and 23 female . The average age was 41 . The samples were taken from the forehead, nose, cheeks, eyelashes and the temporal areas using the standardized skin surface biopsy and hair epilation techniques that were modified for this study . The data obtained were assessed statistically . Demodex was determined in 10% of all the cases . The cases revealed that demodex increased in old age and was more frequently seen in people with fair complexion . It was most frequently found in the forehead and the cheek . The longest postmortem interval in the positive cases was 55 h . The relationship between the postmortem interval and the presence of demodex was not statistically significant . Considering the fact that demodex, which is transmitted from human to human through skin contact, acts as a vector of pathogenic organisms and can stay alive in dead bodies for a long time, we think that the personnel performing autopsies without taking the necessary precautions are under risk.

Genet Sel Evol, 2003, 35 Suppl 1, S83 - 96
Association of the porcine C3 gene with haemolytic complement activity in the pig; Mekchay S et al.; The complement component C3 plays an essential role in the activated complement system, which is involved in phagocytosis, inflammation and immunoregulation to destroy infectious microorganisms . The C3 molecule has more implications in the general defence mechanisms . In this study, the porcine C3 cDNA sequences including 5'- and 3'- flanking regions were determined and the polymorphisms in this gene were identified to carry out an association analysis between C3 and complement activity traits . Porcine C3 gene has high homology with human C3 . Five single nucleotide polymorphisms (SNPs) and one microsatellite were detected in the porcine C3 gene . Haemolytic complement activity of alternative and classical pathways (ACH, CCP) was measured in 416 F2 animals of a crossbred of Duroc x Berlin Miniature Pig, which were immunized with Mycoplasma, Aujeszky and PRRS vaccines . C3 markers were found to be significantly associated (P <0.05) with both ACP and CCP . Animals with the more frequent haplotype present in Duroc and other commercial breeds exhibit higher ACP and CCP levels than the animals with haplotype specific to some Berlin Miniature Pigs . The association of C3 with complement activity reinforces the importance of C3 as a candidate gene for natural resistance to microorganisms.

Water Sci Technol, 2003, 47(12), 191 - 8
Investigation of membrane processes for the removal of volatile fatty acids; Barnes S et al.; The accumulation of volatile fatty acids such as acetic acid can cause reactor pH problems and the inhibition of microorganisms utilised in anaerobic digestion processes . A cross-flow membrane process using Teflon and ion-exchange membranes was investigated as a means of separating acetic acid from pure acetic acid solution and rumen fluid . Acetic acid transfer across the Teflon membrane was dependent on the free acid concentration (CH3COOH) in the acid solution . Concurrent transfer of water was minimal due to the hydrophobic nature of the membrane . The strong base anionic exchange membrane facilitated the separation of acetic acid from both pure solutions and rumen fluid with flux again being dependent on the free acid gradient across the membrane . Flux rates were lower than other studies of diffusion dialysis, however, this may be partly attributed to improper preparation of the membrane . The currently achieved rates of transfer using these membranes are very low and are therefore not yet suitable for full-scale use in anaerobic digestion . Additional research is needed to achieve higher trans-membrane transport rates at reasonable costs.

Cell Microbiol, 2003 Sep, 5(9), 573 - 80
Drosophila blood cells; Meister M et al.; Drosophila blood cells or haemocytes belong to three lineages: plasmatocytes, crystal cells and lamellocytes . There is no equivalent of a lymphoid lineage in insects which have no adaptive immunity . Haematopoiesis is under the control of a number of transcription factors and signalling pathways (such as GATA factors, JAK/STAT or Notch pathways) most of which have homologues which participate in the control of mammalian haematopoiesis . Drosophila plasmatocytes are professional phagocytes reminiscent of the cells from the mammalian monocyte/macrophage lineage . Several receptors responsible for recognition of microorganisms or apoptotic corpses have been identified, which include a Scavenger Receptor, a CD36 homologue and a peptidoglycan recognition protein . Crystal cells contain the enzymes necessary for humoral melanization that accompanies a number of immune reactions . The production of melanin generates, as by-products, cytotoxic free radicals that are believed to participate in the killing of pathogens . Finally, lamellocytes represent a cell type that specifically differentiates after parasitism of Drosophila larvae and forms a capsule around the invader . Encapsulation together with melanization eventually kill the parasite within the capsule.

Ying Yong Sheng Tai Xue Bao, 2003 May, 14(5), 823 - 8
{Advances of research on combined pollution in soil-plant systems}; Guo G et al.; The research on combined pollution in soil-plant systems is a scientific frontier of pollution ecology, and has an important significance in ecological safety of agricultural environment . The origin of combined pollution as a conception and its connotation development were reviewed . Types of combined pollution occurred in soil-plant systems and relevant progresses in this area were summarized . Ecological effects resulted from combined pollution of soil-plant systems and their quantificational tokens were expounded . The interactions of heavy metal-organic pollutant and organic pollutant-pathogenic microorganisms were pointed out to be important combined pollution in soil-plant systems, and the studies on the interactions, sub-products and their aged process, and moletoxicology mechanism of multi-pollutants were suggested to be the future keystone contents in this area . Meanwhile, the prospective studies on the methods and application of the results of combined pollution were put forward to provide a support for the warning and remediation of soil pollution.

Ukr Biokhim Zh, 2002 Nov-Dec, 74(6), 34 - 42
{Analysis of specific nucleotide sequences . DNA biosensors}; Starodub NF et al.; Information about common molecular-biological approaches for the determination of the specific nucleotide sequences in genetic materials was given in the review . Main attention was paid to consideration of the ways for DNA biosensor creation . The information about the types of such biosensors was presented in detail and characteristics of the developed devices were cited . Separately the question about the use of the instrumental analytical approaches for the identification of genetic materials of individual pathogenic microorganisms was viewed.

Rev Argent Microbiol, 2003 Apr-Jun, 35(2), 86 - 90
{Indicators of contamination and their relation to the presence of Escherichia coli in ready-to-eat foods}; de Sousa GB et al.; The relationship between indicator microorganism counts and the presence of Escherichia coli was determined in ready-to-eat food in food stores . Aerobic counts (CA), total coliforms (CT) and molds and yeast (ML) were registered in each food sample as well as the presence of E . coli in food, surface and hand samples . There was a high percentage of E . coli in cooked food (46% in 1 g), in raw food (31% in 0.1 g), in surfaces (37%) and in hands (21%) . Significant correlations were found in CT, CA and ML in cooked food (P = 0.0001); no significant correlations were found in raw food (P > 0.01) . The CT count in cooked food with E . coli was significantly higher than CT count in cooked food without E . coli (median 5.00 cfu/g and 1.54 cfu/g, respectively) . Meanwhile, no significant differences were found in raw food.

Ying Yong Sheng Tai Xue Bao, 2003 Apr, 14(4), 481 - 6
{Phosphorus transfer between mixed poplar and black locust seedlings}; He W et al.; In this paper, the 32P radio-tracer technique was applied to study the ways of phosphorus transfer between poplar (Populus euramericana cv . 'I-214') and black locust (Robinia pseudoacacia) . A five compartment root box (18 cm x 18 cm x 26 cm) was used for testing the existence of the hyphal links between the roots of two tree species when inoculated with vesicular-arbuscular (VA) mycorrhizal fungus (Glomus mosseae) . Populus I-214 (donor) and Robinia pseudoacacia (receiver) were grown in two terminal compartments, separated by a 2 cm root-free soil layer . The root compartments were lined with bags of nylon mesh (38 microns) that allowed the passage of hyphae but not roots . The top soil of a mixed stand of poplar and black locust, autoclaved at 121 degrees C for one hour, was used for growing seedlings for testing . In 5 compartment root box, mycorrhizal root colonization of poplar was 34%, in which VA mycorrhizal fungus was inoculated, whereas 26% mycorrhizal root colonization was observed in black locust, the other terminal compartment, 20 weeks after planting . No root colonization was observed in non-inoculated plant pairs . This indicated that the mycorrhizal root colonization of black locust was caused by hyphal spreading from the poplar . Test of tracer isotope of 32P showed that the radioactivity of the treatment significantly higher than that of the control (P < 0.05), 14 days from the tracer applied, to 27 days after, when VA mycorrhizal fungus was inoculated in poplar root . Furthermore, mycorrhizal interconnections between the roots of poplar and black locust seedlings was observed in situ by binocular in root box . All these experiments showed that the hyphal links was formed between the roots of two species of trees inoculated by VA mycorrhizal fungus . Four treatments were designed according to if there were two nets (mesh 38 microns), 2 cm apart, between the poplar and black locust, and if the soil in root box was pasteurized . Most significant differences of radioactivity among four treatments appeared 44 days after feeding 32P, the radioactivity of the day was applied to estimating the contribution of the various possible transfer ways to the total amount of nutrient transfer . Level of 32P radioactivity was found to be significantly (P < 0.05) higher in leaves of the treatment of "no separated and soil non-pasteurized" (17.1 pulse.g-1.s-1) than in leaves of "net separated and soil non-pasteurized" (5.3 pulse.g-1.s-1), and also significantly higher in leaves of "no net separated and soil pasteurized" (11.5 pulse.g.s-1) than in leaves of "net separated and soil pasteurized" (2.3 pulse.g-1.s-1), and very significantly (P < 0.01) higher in leaves of "no net separated and soil non-pasteurized" than in leaves of "net separated and soil pasteurized", whereas the levels of 32P radioactivity were not significantly different between the other treatments . The results showed that root contact and root exudations were the main ways of phosphorus transfer between the two species and the amount of phosphorus transfer through these two ways accounted for 62% of the total . The activity of the microorganisms including VA mycorrhizal fungi and the interaction between the microorganisms and root contact and root exudations made up 38% of total amount of phosphorus . The effect of mycorrhizal hyphal links in the direct nutrient transfer between poplar and black locust through separate mesh (38 microns) was little.

Anal Bioanal Chem, 2003 Oct, 377(3), 540 - 9 Epub 2003 Aug 12.
Sensor strategies for microorganism detection--from physical principles to imprinting procedures; Dickert FL et al.; Detecting cells and microorganisms in different matrices is becoming an increasingly important task in a variety of fields including bioprocess control, food technology, health care, and environmental analysis . In this review, fast on-line detection methods for this purpose are presented including different recognition and transducer strategies.

Biomol Eng, 2003 Jul, 20(4-6), 261 - 71
Monoseptic cultivation of phototrophic microorganisms--development and scale-up of a photobioreactor system with thermal sterilization; Walter C et al.; The use of phototrophic microorganisms as sources of biological active substances in photoautotrophic and mixotrophic cultivation modes requires an adequate cultivation system with thermal sterilization . A corresponding photobioreactor system in the 10, 25 and 100 l scales was developed . This "Medusa"-photobioreactor system represents a concept based on the air-lift loop principle, whose working volume is irradiated by external light sources . The incident irradiation can be varied by a light control system . An effective CO(2)/O(2) gas exchange is enabled due to the efficient supply with process gas by several gas supply nozzles within the system and a large degassing surface . Using a model to describe the growth characteristics of the organisms, the volumetric irradiation coefficient I(DX) was defined as scale-up parameter . On this basis the scale-up from 1 l bubble columns to the 10 and 100 l scales was realized . The scale-up was performed successfully with Chlorella salina as model organism . A maximum biomass concentration of 7.89 g (dry weight) l(-1) at a maximum specific growth rate of 0.058 h(-1) and a yield of 35 mg l(-1) h(-1) was obtained in a batch cultivation in the 100 l scale under photoautotrophic conditions with an initial biomass concentration of approx . 0.03 g l(-1).

Biomol Eng, 2003 Jul, 20(4-6), 115 - 23
Optimisation of cultivation parameters in photobioreactors for microalgae cultivation using the A-stat technique; Barbosa MJ et al.; Light availability inside the reactor is often the bottleneck in microalgal cultivation and for this reason much attention is being given to light limited growth kinetics of microalgae, aiming at the increase of productivity in photobioreactors . Steady-state culture characteristics are commonly used for productivity optimisation and for cell physiology studies in continuous cultures, and are normally achieved using chemostat cultivations . In the present study, we investigated the applicability of a new and dynamic cultivation method called acceleration-stat (A-stat) to microalgae cultivations where light is the limiting substrate . In the A-stat, the dilution rate is increased at a constant rate . This acceleration rate should be a compromise between a short cultivation time, in order to make it a fast process, and the metabolic adaptation rate of the microorganism to changes in the environment . Simulations of the A-stat were done with different acceleration rates to have an indication of the best rate to use . An A-stat was performed in a pilot plant bubble column (65 l) with Dunaliella tertiolecta as a model organism, and results showed that a pseudo steady state was maintained throughout the experiment . From this work, it was concluded that the A-stat can be used as a fast and accurate tool to determine kinetic parameters and to optimise any specific type of photobioreactor.

Chemosphere, 2003 Oct, 53(3), 217 - 21
Effect of the herbicides oxadiazon and oxyfluorfen on phosphates solubilizing microorganisms and their persistence in rice fields; Das AC et al.; A field experiment has been conducted with two herbicides viz . oxadiazon {5-terbutyl-3-(2,4-dichloro-5-isopropoxyphenyl)-1,3,4-oxadiazol-2-one} and oxyfluorfen {2-chloro-1-(3-ethoxy-4-nitrophenyl)-4-(trifluoromethyl) benzene} at rates of 0.4 and 0.12 kg a.i . ha(-1), respectively, to investigate their effect on the growth and activities of phosphate solubilizing microorganisms in relation to availability of phosphorus as well as persistence of the herbicides in the rhizosphere soil of wetland rice (Oryza sativa L . variety IR-36) . Application of herbicides stimulated the population and activities of phosphate solubilizing microorganisms and also the availability of phosphorus in the rhizosphere soil . Oxyfluorfen provided greater microbial stimulation than oxadiazon . Dissipation of oxyfluorfen and oxadiazon followed first order reaction kinetics with half-life (T(1/2)) of 8.8 and 12 days, respectively . Sixty days after application 0.5% and 3% of the applied oxadiazon and oxyfluorfen residues persisted, respectively, in the rhizosphere soil of rice.

Arch Esp Urol, 2003 Jun, 56(5), 462 - 6
{Genital histoplasmosis . Presentation of a clinical case}; Rodriguez N et al.; OBJECTIVES: On December 1905 Darling, resident histopathologist on the Panama Channel area, observed a parasite in the large mononuclear cells of different organs . Negroni in 1914 described the first Argentinean case . Lung, oral, nasal, and laryngeal involvement are common . On the other hand, genital involvement is uncommon . The objective of this war is to communicate the case of a patient with genital histoplasmosis . METHODS/RESULTS: A 60-year-old Argentinian male started two years before with penile lesions and pruritus . Two partial penile resections had been performed in 1999, but pathology reports were not available . The lesions evolved towards ulceration and bleeding . Surgical toilette was carried out . A partial penectomy and glanuloplasty with an original technique were performed . Pathology revealed intracellular and extracellular spherical microorganisms . Morphologically it was histoplasma capsulatum . The histoplasmosis is an endemic mycosis . The infection takes place when inhaling the microconidiae of the fungus filamentous phase . Only a small number of people develop the disease after infection . Diagnosis is made by discovering of 2-3 micron yeasts with Giemsa stain . CONCLUSIONS: If a patient from the histoplasmosis endemic area has a granular lesion, a mycotic disease should be considered.

Ecotoxicol Environ Saf, 2003 Sep, 56(1), 140 - 7
Issues underlying use of biosensors to measure metal bioavailability; Rensing C et al.; Heavy metal-mediated toxicity in the environment is dependent on bioavailable metal concentrations both internal and external to microbial cells . Both internal and external metal bioavailability are influenced by multiple factors in the soil environment . External factors include pH, redox potential, ionic strength, organic matter and clay content . The internal bioavailable metal concentration is dependent on both the aforementioned external factors, as well as metal uptake and efflux activities that are specific for each microorganism . The metal-specific biosensors discussed in this article can be used to measure internal metal bioavailability.

Ecotoxicol Environ Saf, 2003 Sep, 56(1), 52 - 62
Underlying issues in bioaccessibility and bioavailability: experimental methods; Hund-Rinke K et al.; This article presents experimental designs focusing on assessing of the bioavailability of metals in aquatic organisms, soil organisms, microorganisms, plants, birds, and mammals . Standardized test systems receive the greatest emphasis . With regard to microorganisms, animals, and plants, the study concentrates on toxicity as an indicator for bioavailability . In respective test procedures, results are usually calculated for total chemical concentrations; chemical analyses are not commonly in routine assessments . For soil organisms chiefly exposed by the water pathway, the bioavailable fraction of contaminants can be roughly determined by chemical analyses in aqueous soil extracts simulating soil pore water concentrations . Human toxicity can be determined using adequate in vitro test designs . In addition to experimental designs, results from the literature dealing with specific problems of bioavailability are presented.

J Trauma, 2003 Aug, 55(2), 350 - 4
The pathomechanism of posttraumatic edema of the lower limbs: II--Changes in the lymphatic system; Szczesny G et al.; BACKGROUND: The peripheral lymphatic system reacts to penetrating microorganisms and self-antigens released from tissues and cells damaged by trauma or intracellular pathogens . The response of regional lymph nodes to tissue trauma has not been thoroughly studied . We investigated the changes in lower limb lymphatics and nodes after fractures and soft tissue injuries . This type of injury is frequently complicated by limb edema . Posttraumatic edema of lower limbs is characterized by long-lasting swelling of the limb, erythema, and increased skin temperature at the site of injury . This suggests that a local inflammatory process is proceeding, even though the process of bone or soft tissue healing is considered to be completed . METHODS: Twenty-one patients with closed lower limb bone fractures and soft tissues injuries were studied by means of isotope lymphography . RESULTS: Dilated lymphatics of the entire limb were found in all patients, and 62% of them showed enlarged inguinal lymph nodes . Venous thrombosis was found in 24% of cases . There was no correlation between the degree of lymphatic dilatation, lymph node enlargement, and bone fracture or soft tissue injury or venous thrombosis . Surgical intervention was not an independent factor for lymph node enlargement . CONCLUSION: This study has shown that although the fracture or injured tissues are clinically healed, local inflammatory reaction at the site of injury persists and cytokine signals are sent to the regional lymph nodes.

Crit Rev Ther Drug Carrier Syst, 2003, 20(1), 63 - 102
Ethosomes: new prospects in transdermal delivery; Godin B et al.; Ethosomes are noninvasive delivery carriers that enable drugs to reach the deep skin layers and/or the systemic circulation . Although ethosomal systems are conceptually sophisticated, they are characterized by simplicity in their preparation, safety, and efficacy--a combination that can highly expand their application . Ethosomes are soft, malleable vesicles tailored for enhanced delivery of active agents . This article reviews work carried out in vitro, in vivo, in animal models, and in humans with various ethosomal systems incorporating a wide range of drugs . Because of their unique structure, ethosomes are able to encapsulate and deliver through the skin highly lipophilic molecules such as cannabinoids, testosterone, and minoxidil, as well as cationic drugs such as propranolol and trihexyphenidil . Results obtained in a double-blind two-armed randomized clinical study showed that treatment with the ethosomal acyclovir formulation significantly improved all the evaluated parameters . Preliminary studies with plasmids and insulin revealed that the ethosomal carrier may be used for enhanced delivery of these agents . In further work, the ethosomal technology was broadened to introduce agents into cultured cells and microorganisms . Enhanced delivery of bioactive molecules through the skin and cellular membranes by means of an ethosomal carrier opens numerous challenges and opportunities for the research and future development of novel improved therapies.

J Exp Biol, 2003 Sep, 206(Pt 18), 3149 - 57
Metabolite comparisons and the identity of nutrients translocated from symbiotic algae to an animal host; Whitehead LF et al.; Dinoflagellate algae of the genus Symbiodinium in symbiosis with marine animals release much of their photosynthetic carbon to the animal host . The compounds translocated to the host ('mobile compounds') were investigated by metabolite comparison as follows: a substrate was identified as a candidate mobile compound when comparable profiles of metabolites were generated from host metabolism of this substrate (supplied exogenously) and the endogenous mobile compounds . When the sea anemone Anemonia viridis was incubated with NaH14CO2 under photosynthesizing conditions, most of the radioactivity in the animal tissue was recovered from the low-molecular-mass fraction and distributed in the ratio 1:2:1 between the neutral, acidic and basic sub-fractions . Prominent 14C-labelled compounds included glucose, malate and glucose-6-phosphate . When the symbiosis was incubated with 14C-labelled glucose plus succinate or fumarate (but none of eight other substrate combinations tested), the 14C-labelled metabolites closely matched those obtained with NaH14CO2 . These data suggest that glucose and succinate/fumarate (or metabolically allied compounds) may be important photosynthetic compounds transferred from the Symbiodinium cells to the tissues of A . viridis . Metabolite comparisons can be applied to study nutritional interactions in symbioses involving photosynthetic algae and, with appropriate modification, other associations between microorganisms and plants or animals.

Int J Environ Health Res, 2003 Sep, 13(3), 295 - 302
Comparison of the disinfection efficacy of chlorine-based products for inactivation of viral indicators and pathogenic bacteria in produce wash water; Chaidez C et al.; Outbreaks of pathogenic bacteria infections associated with the consumption of fresh produce has occurred with increased frequency in recent years . This study was undertaken to determine the efficacy of three commonly used disinfectants in packing-houses of Culiacan, Mexico (sodium hypochlorite {NaOCl}, trichlor-s-triazinetrione {TST} and thrichlormelamine {TCM}) for inactivation of viral indicators and pathogenic bacteria inoculated onto produce wash water . Each microbial challenge consisted of 2 L of water containing approximately 8 log10 bacterial CFU ml(-1), and 8 log10 viral PFU ml(-1) treated with 100 and 300 mg l(-1) of total chlorine with modified turbidity . Water samples were taken after 2 min of contact with chlorine-based products and assayed for the particular microorganisms . TST and NaOCl were found to effectively reduce for bacterial pathogens and viral indicators 8 log10 and 7 log10, respectively (alpha=0.05) . The highest inactivation rate was observed when the turbidity was low and the disinfectant was applied at 300 mg l(-1) . TCM did not show effective results when compared with the TST and NaOCl (P<0.05) . These findings suggest that turbidity created by the organic and inorganic material present in the water tanks carried by the fresh produce may affect the efficacy of the chlorine-based products.

AIHA J (Fairfax, Va), 2003 Jul-Aug, 64(4), 487 - 95
Exposure to particulates, microorganisms, beta(1-3)-glucans, and endotoxins during soybean harvesting; Roy CJ et al.; Sclerotinia sclerotiorum is an emerging fungal pathogen affecting soybeans in the United States . In response to its emergence, exposures to particulates, bioaerosols, endotoxins, S . sclerotiorum, and beta(1-3)-glucans were characterized during soybean harvesting . Air sampling was performed on soybean harvesters (combines) and on the farmers in closed cabs as personal samples during harvesting at 17 farms in 1997 and repeated at 15 in 1998 . S . sclerotiorum infestation was evident in the fields at 8 of the sites (44%) . The geometric mean concentrations (and geometric standard deviations) measured on the combines in 1998 were as follows: total dust, 11.9 (2.8) mg/m(3); inhalable dust 11.7 (6.4) mg/m(3); and beta(1-3)-glucans, 5027 (7) ng/m(3) . Values for the personal samples in 1998 were as follows: total dust, 1.2 (6.7) mg/m(3); inhalable dust, 1.1 (5.3); and beta(1-3)-glucans, 674 (9) ng/m(3) . These concentrations were two- to threefold higher than in the previous year . Ambient endotoxin concentrations were 4438 EU/m(3) in Year I and 459 EU/m(3) in Year II . Particle size distribution measurements on the combines yielded mass median aerodynamic diameters of 6.6 microm on the combine and 4.0 microm inside the combine cab . Closed combine cabs provided an average workplace protection factor of 11.7 for total dust . Nevertheless, personal exposures to organisms inside combine cabins ranged from 3.6 x 10(4) to 4.0 x 10(8) organisms/m(3) . These data indicate the potential exists for high exposures to organic dust and bioaerosols during soybean harvesting.

Crit Care Med, 2003 Aug, 31(8 Suppl), S524 - 31
Injury and repair in lung and airways; Shimabukuro DW et al.; Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are common causes of morbidity and mortality in the intensive care unit . ALI/ARDS occurs as a result of systemic inflammation, usually triggered by a microorganism . Activation of leukocytes and release of proinflammatory mediators from multiple cellular sources result in both local and distant tissue injury . Tumor necrosis factor-alpha and interleukin-1 beta are the best characterized of the proinflammatory cytokines contributing to ALI/ARDS and subsequent fibrosis . The ultimate clinical course of ALI/ARDS often is determined by the ability of the injured lung to repopulate the alveolar epithelium with functional cells . Death may occur when fibrosis predominates the healing response, as it results in worsening lung compliance and oxygenation . The rodent bleomycin model of lung fibrosis allows the use of molecular tools to dissect the cellular and subcellular processes leading to fibrosis . The elements of this response may provide therapeutic targets for the prevention of this devastating complication of ALI/ARDS.

Ig Sanita Pubbl, 2002 Jan-Apr, 58(1-2), 75 - 88
{Etiology of rare diseases caused by novel viral agents: a review of the most recent literature}; Prota M et al.; The authors describe the viral pathologies that have become rare in Italy and in the rest of the western world; they also analyze some novel agents mainly imported from the tropical and Asian Countries . For some of the microorganisms, they describe the various zones at risk, ways of transmission, clinical case, recent lab methods and preventive measures.

Ann Clin Microbiol Antimicrob . 2003 Jul 16;2(1):7.
Prescribing patterns of antibiotics and sensitivity patterns of common microorganisms in the Internal Medicine ward of a teaching hospital in Western Nepal: a prospective study; Shankar RP et al.; BACKGROUND: Information about antibiotic use and resistance patterns of common microorganisms are lacking in hospitals in Western Nepal . Excessive and inappropriate use of antibiotics contributes to the development of bacterial resistance . The parameter: Defined daily dose/100 bed-days, provides an estimate of consumption of drugs among hospital in-patients . This study was carried out to collect relevant demographic information, antibiotic prescribing patterns and the common organisms isolated including their antibiotic sensitivity patterns . METHODS: The study was carried out over a 3-month period (01.04.2002 to 30.06.2002) at the Manipal Teaching Hospital, Western Nepal . The median number of days of hospitalization and mean PlusMinus; SD cost of antibiotics prescribed during hospital stay were calculated . The use of antibiotics was classified for prophylaxis, bacteriologically proven infection or non-bacteriologically proven infection . Sensitivity patterns of the common organisms were determined . Defined daily dose/100 bed-days of the ten most commonly prescribed antibiotics were calculated . RESULTS: 203 patients were prescribed antibiotics; 112 were male . Median duration of hospitalization was 5 days . 347 antibiotics were prescribed . The most common were ampicillin, amoxicillin, metronidazole, ciprofloxacin and benzylpenicillin . Mean PlusMinus; SD cost of antibiotics was 16.5 PlusMinus; 13.4 US$ . Culture and sensitivity testing was carried out in 141 patients . The common organisms isolated were H . influenzae, E . coli, K . pneumoniae and S . aureus . CONCLUSIONS: Antibiotic resistance is becoming a problem in the Internal Medicine ward . Formulation of a policy for hospital antibiotic use and an educational programme especially for junior doctors is required.

J Agric Food Chem, 2003 Aug 13, 51(17), 4866 - 72
Differentiation of natural and synthetic phenylalanine and tyrosine through natural abundance 2H nuclear magnetic resonance; Brenna E et al.; The natural abundance deuterium NMR characterization of samples of the amino acids tyrosine (1) and phenylalanine (2), examined as the acetylated methyl esters 4 and 6, has been performed with the aim to identify by these means the contribution in animals of the hydroxylation of the diet l-phenylalanine (2) to the formation of l-tyrosine (1), a feature previously revealed on the same samples through the determination of the phenolic delta(18)O values . The study, which includes also the NMR examination of benzoic acid (5) from 2 and of tyrosol (7) from 1, substantially fails in providing the required information because the mode of deuterium labeling of tyrosine samples of different origins is quite similar but indicates a dramatic difference in the deuterium labeling pattern of the two amino acids 1 and 2 . The most relevant variation is with regard to the deuterium enrichments at the CH(2) and CH positions, which are inverted in the two amino acids of natural derivation . Moreover, whereas the diastereotopic benzylic hydrogen atoms of l-tyrosine (1) appear to be equally deuterium enriched, in l-phenylalanine (2) the (D/H)(3)(R)() > (D/H)(3)(S)() . Similarly, benzoic acid (5) shows separate signals for the aromatic deuterium nuclei, which are quite indicative of the natural or synthetic derivation . The mode of deuterium labeling of the side chain of 1 and 2 is tentatively correlated to the different origins of the two amino acids, natural from animal sources for l-tyrosine and biotechnological probably from genetically modified microorganisms for l-phenylalanine.

Kisaengchunghak Chapchi, 1983 Dec, 21(2), 193 - 204
{Distribution Of Medically Important Freshwater Snails And Larval Trematodes From Parafossarulus Manchouricus And Semisulcospira Libertina Around The Jinyang Lake In Kyongsang-Nam-Do, Korea}; Cho HC et al.; The Jinyang Lake is dammed up Nam River, and surrounded by Jinju city and four counties; Jinyang, Sanchong, Hadong, and Sachon in Kyongsang-Nam-Do, Korea . The area around this man-made lake have been known as an endemic focus of clonorchiasis in Korea . The present study was first aimed to know the distribution of freshwater mollusks including medically important snails, and larval trematodes shed from Parafossarulus manchouricus and Semisulcospira libertina . In addtion to above studies, water analyses in each snail habitat were carried out in order to figure out a part of their environmental factors . This malaco-ecological survey was done at the six areas around upper, middle and lower parts of the lake for 4 months, August-November, 1983 . Total nine species of freshwater mollusks were collected throughout the study: 4 species of gastropods; Semisulcospira libertina, Cipangopaludina chinensis, Parafossarulus manchouricus and Radix auricularia, and 5 species of bivalves; Unio douglasiae, Anodonta woodiana, Lamprotula gottschei, Corbicula fluminea and Limnoperma lucustris . Out of nine species of freshwater mollusks, three species of gastropods; S . libertina, P . manchouricus and R . auricularia were medically important in terms of the transmission of digenetic trematodes to humans . P . manchouricus and R . auricularia were mainly collected from the shallow ponds and the irrigation channels with the muddy basin, but S . libertina and the bivalves were only collected from the stream of Nam river where the gravels and rocks were dominant . The levels of dissolved oxygen(D.O.) and biochemical oxygen demand (B.O.D.(5)) of the water specimens sampled from the study areas ranged from 6.0 to 9.6 ppm and from 0.4 to 1.6 ppm respectively . As a result, it is considered that water system around the Jinyang Lake might be relatively clean without any heavy pollution of aquatic microorganisms and organic materials during the period of this study . On the other hand, eight metalic constituents from the water samples were also assayed, and all metalic ions detected were remarkably low below the legal criteria . However, calcium ion in the water samples from the habitats of P . manchouricus was considerably higher than others . Infection rates of digenetic trematodes in the snails were 6.9 % in P . manchouricus and 4.8 poercent in S . libertina, respectively . P . manchouricus snails harboring with the cercariae of Clonorchis sinensis were only 0.14 % among the snails examined and other trematode cercariae except cercaria of C . sinensis were; furcocercus cercariae, cercaria of Loxogenes liberum type I and II . S . libertina snails parasitized with the cercariae of Metagonimus yokogawai were 1.5 % out of the snails examined and no cercaria of Paragonimus westermani was found in S . libertina snails in the present study . Digenetic trematode cercariae other then M . yokogawai in S . libertina snails were: Cercaria yoshidae (B type), Cercaria cristata, Cercaria innominatum, Cercaria of Centrocestus formosanus and Cercaria nipponensis.

Appl Environ Microbiol, 2003 Aug, 69(8), 5011 - 4
Effects of essential oils on ruminal microorganisms and their protein metabolism; McIntosh FM et al.; A commercial blend of essential oil (EO) compounds was added to a grass, maize silage, and concentrate diet fed to dairy cattle in order to determine their influence on protein metabolism by ruminal microorganisms . EO inhibited (P < 0.05) the rate of deamination of amino acids . Pure-culture studies indicated that the species most sensitive to EO were ammonia-hyperproducing bacteria and anaerobic fungi.

Appl Environ Microbiol, 2003 Aug, 69(8), 4892 - 900
Single-leaf resolution of the temporal population dynamics of Aureobasidium pullulans on apple leaves; Woody ST et al.; The abundance of phylloplane microorganisms typically varies over several orders of magnitude among leaves sampled concurrently . Because the methods traditionally used to sample leaves are destructive, it has remained unclear whether this high variability is due to fixed differences in habitat quality among leaves or to asynchronous temporal variation in the microbial population density on individual leaves . We developed a novel semidestructive assay to repeatedly sample the same apple leaves from orchard trees over time by removing progressively more proximal approximately 1-cm-wide transverse segments . Aureobasidium pullulans densities were determined by standard leaf homogenization and plating procedures and were expressed as CFU per square centimeter of segment . The A . pullulans population densities among leaves were lognormally distributed . The variability in A . pullulans population densities among subsections of a given leaf was one-third to one-ninth the variability among whole leaves harvested concurrently . Sequential harvesting of leaf segments did not result in detectable changes in A . pullulans density on residual leaf surfaces . These findings implied that we could infer whole-leaf A . pullulans densities over time by using partial leaves . When this successive sampling regimen was applied over the course of multiple 7- to 8-day experiments, the among-leaf effects were virtually always the predominant source of variance in A . pullulans density estimates . Changes in A . pullulans density tended to be synchronous among leaves, such that the rank order of leaves arrayed with respect to A . pullulans density was largely maintained through time . Occasional periods of asynchrony were observed, but idiosyncratic changes in A . pullulans density did not contribute appreciably to variation in the distribution of populations among leaves . This suggests that persistent differences in habitat (leaf) quality are primarily responsible for the variation in A . pullulans density among leaves in nature.

Appl Environ Microbiol, 2003 Aug, 69(8), 4806 - 13
Targeting single-nucleotide polymorphisms in the 18S rRNA gene to differentiate Cyclospora species from Eimeria species by multiplex PCR; Orlandi PA et al.; Cyclospora cayetanensis is a coccidian parasite that causes protracted diarrheal illness in humans . C . cayetanensis is the only species of this genus thus far associated with human illness, although Cyclospora species from other primates have been named . The current method to detect the parasite uses a nested PCR assay to amplify a 294-bp region of the small subunit rRNA gene, followed by restriction fragment length polymorphism (RFLP) or DNA sequence analysis . Since the amplicons generated from C . cayetanensis and Eimeria species are the same size, the latter step is required to distinguish between these different species . The current PCR-RFLP protocol, however, cannot distinguish between C . cayetanensis and these new isolates . The differential identification of such pathogenic and nonpathogenic parasites is essential in assessing the risks to human health from microorganisms that may be potential contaminants in food and water sources . Therefore, to expand the utility of PCR to detect and identify these parasites in a multiplex assay, a series of genus- and species-specific forward primers were designed that are able to distinguish sites of limited sequence heterogeneity in the target gene . The most effective of these unique primers were those that identified single-nucleotide polymorphisms (SNPs) at the 3' end of the primer . Under more stringent annealing and elongation conditions, these SNP primers were able to differentiate between C . cayetanensis, nonhuman primate species of Cyclospora, and Eimeria species . As a diagnostic tool, the SNP PCR protocol described here presents a more rapid and sensitive alternative to the currently available PCR-RFLP detection method . In addition, the specificity of these diagnostic primers removes the uncertainty that can be associated with analyses of foods or environmental sources suspected of harboring potential human parasitic pathogens.

Appl Environ Microbiol, 2003 Aug, 69(8), 4403 - 7
Elucidation of the transmission patterns of an insect-borne bacterium; Darby AC et al.; Quantitative data on modes of transmission are a crucial element in understanding the ecology of microorganisms associated with animals . We investigated the transmission patterns of a gamma-proteobacterium informally known as pea aphid Bemisia-like symbiont (PABS), also known as T-type, which is widely but not universally distributed in natural populations of the pea aphid, Acyrthosiphon pisum . The vertical transmission of PABS to asexual and sexual morphs and sexually produced eggs was demonstrated by a diagnostic PCR-based assay, and the maximum estimated failure rate was 2% . Aphids naturally lacking PABS acquired PABS bacteria administered via the diet, and the infection persisted by vertical transmission for at least three aphid generations . PABS was also detected in two of five aphid honeydew samples tested and in all five siphuncular fluid samples tested but in none of 15 samples of salivary secretions from PABS-positive aphids . However, PABS-negative aphids did not acquire PABS when they were cocultured with PABS-positive aphids; the maximal estimated level of horizontal transmission was 18% . A deterministic model indicated that the force of infection by a horizontal transmission rate of 3% is sufficient to maintain a previously described estimate of the prevalence of PABS-positive aphids (37%), if the vertical transmission rate is 98% . We concluded that PABS infections in A . pisum can be maintained by high vertical transmission rates and occasional horizontal transmission, possibly via the oral route, in the absence of selection either for or against aphids bearing this bacterium.

Mikrobiologiia, 2003 May-Jun, 72(3), 301 - 7
{Terminal oxidases in different genera of the family Microbacteriaceae}; Trutko SM et al.; The nature of terminal oxidases in representatives of four different genera of the family Microbacteriaceae was studied . It was found that the late-logarithmic and early-stationary cells of all of the investigated strains of the genera Plantibacter and Okibacterium contain the aa3-type cytochrome oxidase . Bacteria of the genera Rathayibacter and Agreia synthesize three oxidases, the bb3- and aa3-type cytochrome oxidases and nonheme cyanide-resistant oxidase, in proportions dependent on the cultivation conditions and the growth phase . Oxygen deficiency in the cultivation medium induces the synthesis of the bd-type oxidase in all of the microorganisms studied . The data obtained provide evidence that the type of terminal oxidases, along with the known chemotaxonomic characteristics, may serve to differentiate the genera of the family Microbacteriaceae at the phenotypic level.

Chem Rec, 2003, 3(3), 158 - 71
Microbial genes and enzymes in the degradation of chlorinated compounds; Ogawa N et al.; Microorganisms are well known for degrading numerous natural compounds . The synthesis of a multitude of chlorinated compounds by the chemical industry and their release into the natural environment have created major pollution problems . Part of the cause of such pollution is the inability of natural microorganisms to efficiently degrade synthetic chlorinated compounds . Microorganisms are, however, highly adaptable to changes in the environment and have consequently evolved the genes that specify the degradation of chlorinated compounds to varying degrees . Highly selective laboratory techniques have also enabled the isolation of microbial strains capable of utilizing normally recalcitrant highly chlorinated compounds as their sole source of carbon and energy . The evolution and role of microbial genes and enzymes, as well as their mode of regulation and genetic interrelationships, have therefore been the subjects of intense study . This review emphasizes the genetic organization and the regulation of gene expression, as well as evolutionary considerations, regarding the microbial degradation of chlorobenzoates, chlorocatechols, and chlorophenoxyacetic acids .

Dig Surg, 2003, 20(5), 415 - 20 Epub 2003 Jul 31.
Treatment of secondary peritonitis: is a less expensive broad-spectrum antibiotic as effective as a carbapenem?
Genne D, Menetrey A, Jaquet A, Indino P, Senechaud C, Siegrist HH.
BACKGROUND: In this retrospective study, the possibility of using amoxicillin/clavulanate for empirical therapy of secondary peritonitis as an alternative to imipenem was explored . METHODS: All secondary peritonitis cases at our institution between 1998 and 2000 were included . Susceptibility to imipenem and amoxicillin/clavulanate of microorganisms isolated in peritoneal fluid and success rates were compared . Therapeutic failure was defined as death, necessity of repeated surgical intervention, or clinical deterioration with persistent positive cultures . RESULTS: Seventy-six cases of secondary peritonitis with 156 microorganisms were found . One hundred and forty-nine (98%) were susceptible to imipenem versus 124 (82%) to amoxicillin/clavulanate (p = 0.0001) . Thirteen therapeutic failures occurred in 52 patients treated with amoxicillin/clavulanate (25%) versus 3 out of 8 (38%) with imipenem (p = 0.43) . The proportion of organisms resistant to amoxicillin/clavulanate in therapeutic failures was greater in nosocomial versus community-acquired secondary peritonitis (p = 0.041) . CONCLUSION: Despite its better in vitro bacteriological activity, clinical efficacy of imipenem was identical to amoxicillin/clavulanate . The use of amoxicillin/clavulanate instead of imipenem would save 889 Euro per case .

Curr Opin Immunol, 2003 Aug, 15(4), 402 - 7
Toll-like receptors and innate antiviral responses; Vaidya SA et al.; Toll-like receptors (TLRs) have a unique and important role in detecting the presence of pathogenic infection . TLRs can recognize conserved structures from a wide variety of microorganisms, such as bacteria, mycobacteria, spirochetes and yeast . However, they are generally not thought to play a major role in viral infection . Several reports have now identified distinct viral ligands for the TLRs, and evidence is accumulating for a functional role of the TLRs in mediating antiviral effector mechanisms.

FEMS Microbiol Lett, 2003 Aug 8, 225(1), 69 - 73
Construction of a genetically engineered microorganism for CO2 fixation using a Rhodopseudomonas/Escherichia coli shuttle vector; Du C et al.; The CO2 fixation ability of Rhodopseudomonas palustris DH was enhanced by introducing the recombinant plasmid pMG-CBBM containing the form II ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) gene (cbbM) isolated from Rps . palustris NO . 7 . Sequencing of a 3.0-kb PstI fragment containing the cbbM gene revealed an open reading frame encoding 461 amino acids, homologous to known cbbM genes, with a ribosome binding site upstream of cbbM and a terminator downstream of cbbM, without promoter . pMG-CBBM, a Rhodopseudomonas/Escherichia coli shuttle expression plasmid, was derived from the Rhodopseudomonas/E . coli shuttle cloning vector pMG105, by inserting the promoter of the pckA gene and the cbbM gene into its multiple cloning site . Plasmid pMG-CBBM was transformed into Rps . palustris DH by electroporation, and was stably maintained when transformants were grown either photoheterotrophically or photolithoautotrophically in the absence of antibiotics . This is the first report of an expression plasmid containing a Rps . palustris-specific promoter that allows stable expression of a foreign gene in the absence of antibiotic selection.

Chemistry, 2003 Aug 4, 9(15), 3604 - 10
Cofactor recycling mechanism in asymmetric biocatalytic reduction of carbonyl compounds mediated by yeast: which is the efficient electron donor?
Zhang BL, Pionnier S.
In asymmetric reduction of carbonyl compounds mediated by microorganisms, the cofactors that transfer hydride should be regenerated by using a recycling system . In most cases, this recycling system consists of carbohydrate molecules, especially glucose or sucrose . Other molecules such as ethanol and acetate have been used as electron donors too . The reduction can even be conducted without added electron donors . To improve biocatalytic synthesis, it is important to understand the cofactor recycling mechanism . In this work, the hydride-transfer mechanism in cofactor regeneration, which takes place in bioreduction mediated by yeast, was studied by means of an isotope tracing technique . The results show that, when glucose was used, the NADH involved in the glycolysis was consumed directly in the formation of ethanol and was not used in the bioreduction . Hence, the regeneration of cofactors in the reduction is not coupled with glycolysis . Nevertheless, glucose is an efficient electron donor that transfers hydride through the hexose monophosphate (HMP) pathway in which the main hydrogen source is C-1 and C-3 hydrogen of glucose . Ethanol is not a good electron donor, since, when it was used, only a small quantity of hydrogen was transferred from this molecule, and the main hydrogen source was water . Therefore, the ethanol oxidation pathway may not be efficient . In the absence of added auxiliary substrates, the yeast cells may use electron donors stored in its cellules . However, in this case we observed that the main hydrogen source for cofactor recycling was water, while only very few hydrogen atoms were from unexchangeable sites . This is similar to the case in which ethanol is used, and is in contradiction with the HMP pathway if stored glucose was the electron donor . The question that remains to be investigated is "what is the efficient electron donor recycling mechanism in the yeast cellules?"

Funct Integr Genomics, 2003 Dec, 3(4), 160 - 70 Epub 2003 Jul 30.
Global nutritional profiling for mutant and chemical mode-of-action analysis in filamentous fungi; Tanzer MM et al.; We describe a method for gene function discovery and chemical mode-of-action analysis via nutrient utilization using a high throughput Nutritional Profiling platform suitable for filamentous microorganisms . We have optimized the growth conditions for each fungal species to produce reproducible optical density growth measurements in microtiter plates . We validated the Nutritional Profiling platform using a nitrogen source utilization assay to analyze 21 Aspergillus nidulans strains with mutations in the master nitrogen regulatory gene, areA . Analysis of these data accurately reproduced expected results and provided new data to demonstrate that this platform is suitable for fine level phenotyping of filamentous fungi . Next, we analyzed the differential responses of two fungal species to a glutamine synthetase inhibitor, illustrating chemical mode-of-action analysis . Finally, a comparative phenotypic study was performed to characterize carbon catabolite repression in four fungal species using a carbon source utilization assay . The results demonstrate differentiation between two Aspergillus species and two diverse plant pathogens and provide a wealth of new data on fungal nutrient utilization . Thus, these assays can be used for gene function and chemical mode-of-action analysis at the whole organism level as well as interspecies comparisons in a variety of filamentous fungi . Additionally, because uniform distribution of growth within wells is maintained, comparisons between yeast and filamentous forms of a single organism can be performed.

Appl Microbiol Biotechnol, 2003 Dec, 63(2), 222 - 30 Epub 2003 Aug 01.
Polychlorinated biphenyl (PCB) degradation and persistence of a gfp-marked Ralstonia eutropha H850 in PCB-contaminated soil; Abbey AM et al.; Ralstonia eutropha H850 was labelled chromosomally with a gfp marker gene encoding for the green fluorescent protein, and designated R . eutropha H850g13 . Visual observation of green fluorescent cells under an epifluorescence microscope, and PCR amplification products, confirmed that the bacterium was labelled with gfp . Southern blot hybridization products further confirmed the gfp was chromosomally labelled . Using resting cell assays, it was determined that insertion of the gfp gene decreased the microorganisms' ability to degrade biphenyl compared to the parent strain . However, this marker facilitated the identification and monitoring of R . eutropha H850g13 survival in soil microcosm experiments . Survival and polychlorinated biphenyl degradation by R . eutropha H850g13 was analysed in soil microcosms spiked with 2,2',5,5'-tetrachlorobiphenyl (TeCB) . R . eutropha H850g13 was detected by viable plate counts and most-probable-number/PCR after 102 days in TeCB-contaminated soil microcosms, and was likely outcompeted by indigenous soil microorganisms in microcosms amended with oil and Daramend (an organic amendment, R . eutropha H850g13 did not degrade TeCB in any of the soil microcosms . This research confirmed that gfp was useful as a marker to distinguish R . eutropha H850g13 from indigenous soil microorganisms over a 102 day period and that, under the experimental conditions used, R . eutropha H850g13 did not degrade TeCB.

J Clin Invest, 2003 Aug, 112(3), 389 - 97
Critical role of galectin-3 in phagocytosis by macrophages; Sano H et al.; Galectin-3 is a member of a large family of animal lectins . This protein is expressed abundantly by macrophages, but its function in this cell type is not well understood . We have studied the effect of galectin-3 gene targeting on phagocytosis, a major function of macrophages . Compared with wild-type macrophages, galectin-3-deficient (gal3-/-) cells exhibited reduced phagocytosis of IgG-opsonized erythrocytes and apoptotic thymocytes in vitro . In addition, gal3-/- mice showed attenuated phagocytic clearance of apoptotic thymocytes by peritoneal macrophages in vivo . These mice also exhibited reduced IgG-mediated phagocytosis of erythrocytes by Kupffer cells in a murine model of autoimmune hemolytic anemia . Additional experiments indicate that extracellular galectin-3 does not contribute appreciably to the phagocytosis-promoting function of this protein . Confocal microscopic analysis of macrophages containing phagocytosed erythrocytes revealed localization of galectin-3 in phagocytic cups and phagosomes . Furthermore, gal3-/- macrophages exhibited a lower degree of actin rearrangement upon Fcgamma receptor crosslinkage . These results indicate that galectin-3 contributes to macrophage phagocytosis through an intracellular mechanism . Thus, galectin-3 may play an important role in both innate and adaptive immunity by contributing to phagocytic clearance of microorganisms and apoptotic cells.

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