Microorganisms

Water Res, 2003 Feb, 37(3), 539 - 50
Treatment of atrazine in nursery irrigation runoff by a constructed wetland; Runes HB et al.; To investigate the treatment capability of a surface flow wetland at a container nursery near Portland, Oregon, atrazine was introduced during simulated runoff events . Treatment efficiency was evaluated as the percent atrazine recovered (as percent of applied) in the water column at the wetland's outlet . Atrazine treatment efficiency at the outlet of the constructed wetland during a 7-d period ranged from 18-24% in 1998 (experiments 1-3) and 16-17% in 1999 (experiments 4 and 5) . Changes in total flow, or frequency and intensity of runoff events did not affect treatment . For experiment 6 in 1999, where the amount, frequency, and duration of runoff events exceeded all other experiments, treatment was compromised . For all experiments, deethylatrazine (DEA) and deisopropylatrazine (DIA) accounted for 13-21% of the initial application . Hydroxyatrazine (HA) was rarely detected in the water . Organic carbon adsorption coefficients (Koc) were determined from batch equilibrium sorption isotherms with wetland sediment, and they decreased in the order of HA > DIA > atrazine > DEA . Static water-sediment column experiments indicated that sorption is an important mechanism for atrazine loss from water passing through the constructed wetland . The results of the MPN assay indicated the existence in the wetland of a low-density population of microorganisms with the potential to mineralize atrazine's ethyl side chain.

Shock, 2003 Apr, 19(4), 305 - 9
Relationships of circulating nitrite/nitrate levels to severity and multiple organ dysfunction syndrome in systemic inflammatory response syndrome; Mitaka C et al.; Excessive nitric oxide (NO) production has been implicated to be responsible for the development of septic shock . To determine whether plasma nitrite/nitrate (NOx) levels are related to the severity of systemic inflammatory response syndrome (SIRS) and the degree of multiple organ dysfunction, we studied plasma NOx levels in 70 patients with SIRS consisting of noninfectious SIRS (n = 32), sepsis (n = 23), and septic shock (n = 15) . Infection is a microbial phenomenon characterized by an inflammatory response to the presence of microorganism . Positive culture for microorganism is regarded as infectious SIRS (sepsis and septic shock) and negative culture is regarded as noninfectious SIRS . Plasma samples collected from each patient within 24 h from admission to the intensive care unit were subjected for measurement of NOx levels, the stable end products of NO, by the high performance liquid chromatography-Greiss system . Mean plasma NOx levels in patients with SIRS were 52.8 +/- 44 microM/L, ranging from 8.1 to 186.2 microM/L . Plasma NOx levels were positively correlated with Acute Physiology, Age, and Chronic Health Evaluation (APACHE) III score (r = 0.414, P < 0.01) and sequential organ failure assessment (SOFA) score (r = 0.433, P < 0.01) . Plasma NOx levels in patients with sepsis (51.0 +/- 38.5 microM/L) and septic shock (94.5 +/- 53.7 microM/L) were significantly (P < 0.01) higher than those in patients with noninfectious SIRS (25.8 +/- 16.9 microM/L) and healthy subjects (29.6 +/- 8.9 microM/L) . Our study shows that plasma NOx levels are increased in patients with infectious, but not noninfectious SIRS, which increase as the severity of SIRS and the development of multiple organ dysfunction syndrome, suggesting its possible pathogenic role in SIRS.

Chemosphere, 2003 Feb, 50(6), 771 - 4
Rapid degradation of butachlor in wheat rhizosphere soil; Yu YL et al.; The degradative characteristics of butachlor in non-rhizosphere, wheat rhizosphere, and inoculated rhizosphere soils were measured . The rate constants for the degradation of butachlor in non-rhizosphere, rhizosphere, and inoculated rhizosphere soils were measured to be 0.0385, 0.0902, 0.1091 at 1 mg/kg, 0.0348, 0.0629, 0.2355 at 10 mg/kg, and 0.0299, 0.0386, 0.0642 at 100 mg/kg, respectively . The corresponding half-lives for butachlor in the soils were calculated to be 18.0, 7.7, 6.3 days at 1 mg/kg, 19.9, 11.0, 2.9 days at 10 mg/kg, and 23.2, 18.0, 10.8 days at 100 mg/kg, respectively . The experimental results show that the degradation of butachlor can be enhanced greatly in wheat rhizosphere, and especially in the rhizosphere inoculated with the bacterial community designated HD which is capable of degrading butachlor . It could be concluded that rhizosphere soil inoculated with microorganisms-degrading target herbicides is a useful pathway to achieve rapid degradation of the herbicides in soil.

Chemosphere, 2003 Feb, 50(6), 755 - 61
Chemical behavior of Cd in rice rhizosphere; Lin Q et al.; Chemical behavior of Cd in rice rhizosphere as affected or not by Pb was investigated . The NH4OAc extractable Cd in the rhizosphere was distinctly lower than that in bulk soil . The depletion of Cd in the rhizosphere could not be simply attributed to Cd uptake by rice . The observed phenomena could be attributed to the decreasing pH in the rhizosphere and the complexing capabilities of soluble exudates for Cd . Extractable Cd increased in both the rhizosphere and bulk soil after the addition of Pb, which might be caused by the replacement of Pb for Cd . The extractable Cd in the non-rhizosphere varied with the distance from the root surface, especially within 0-1 mm, which was greatly affected by the combined effects of mass flow, activation and fixation, and had the lowest extractable Cd . Pb addition affected the distribution of extractable Cd in the non-rhizosphere, implying that the affinity of Pb for organic matter was greater than that of Cd . The difference of Cd species between rhizosphere and bulk soil demonstrated that the transformation of exchangeable Cd (EXC-Cd) to OM-Cd (bound to organic matter) and FMO-Cd (bound to iron and manganese oxide) occurred in the rice rhizosphere due to the exudations from the rice root, the activity of microorganisms on the root surface and the activation of Fe and Mn oxides . The interaction between Pb and Cd resulted in the content of EXC-Cd being higher in the presence of Pb, whereas the OM-Cd content was lower in the presence of Pb.

J Clin Microbiol, 2003 Apr, 41(4), 1722 - 5
New filamentous fungus Sagenomella chlamydospora responsible for a disseminated infection in a dog; Gene J et al.; A filamentous fungus that caused a fatal systemic infection in a dog has been identified as the new species Sagenomella chlamydospora . When the case was initially reported, the fungus was identified as Paecilomyces sp . This study emphasizes how difficult can be the identification of the causative agent of an infection when an uncommon microorganism is involved . This is the first time that this genus has been involved in animal infections, including humans.

J Clin Microbiol, 2003 Apr, 41(4), 1656 - 63
Heterogeneity of RNA polymerase gene (rpoB) sequences of Mycobacterium gordonae clinical isolates identified with a DNA probe kit and by conventional methods; Itoh S et al.; In a previous study, we have evaluated genetic identification by using the rpoB gene, which was recently introduced by Kim et al . (J . Clin . Microbiol . 39:2102-2109, 2001; J . Clin . Microbiol . 37:1714-1720, 1999) . In this process, we examined the rpoB gene heterogeneity of clinical isolates identified as Mycobacterium gordonae with the conventional biological and biochemical tests and/or a commercially available DNA probe kit . Sequencing of the rpoB gene of 34 clinical isolates revealed that M . gordonae clinical isolates were classified into four major clusters (A, B, C, and D) . Interestingly, organisms belonging to cluster D (15 isolates) did not hybridize with M . gordonae ATCC 14470 and specifically possessed urease activity . Therefore, it could be considered to be a novel mycobacterium . The identification of M . gordonae is known to have ambiguous results sometimes . On the other hand, identification of clinical isolates seems to be inconvenient and unsuitable because of a more than 99% 16S rRNA gene similarity value between clusters . These findings suggest that the existence of M . gordonae-like mycobacteria that share similar biochemical and biological characteristics with the 16S rRNA gene of an M . gordonae type strain but less similarity at the genomic DNA level may have complicated the identification of M . gordonae in many laboratories . Furthermore, compared with hsp65 PCR restriction analysis (PRA), rpoB PRA would have the advantage of producing no ambiguous results because of the intracluster homogeneity of the rpoB gene . In this case, rpoB would provide clearer results than hsp65, even if PRA analysis was used . We demonstrated that these M . gordonae-like mycobacteria were easily distinguished by PRA of the rpoB sequence . Additionally, the significance of this M . gordonae-like cluster may help to establish the comparison between the M . gordonae isolates from a clinical specimen and an infectious process in a given patient and to determine the true incidence of infection with this microorganism.

Environ Sci Technol, 2003 Mar 15, 37(6), 1083 - 8
Analysis of pesticide residues on museum objects repatriated to the Hupa tribe of California; Palmer PT et al.; In the past, it was common practice for museum professionals and private collectors to apply a variety of pesticide agents to objects in their collections to preserve them from depredations by microorganisms, fungi, and other pests . The Native American Graves Repatriation and Protection Act allows federally recognized tribes to request that museums return objects taken from their ancestors . Given that poor records were kept on the treatment of individual objects, it is unknown whether specific objects are contaminated with these pesticide agents . Although chemical analysis represents the only reliable means to determine the types and levels of pesticides on these objects, surprisingly few publications document the extent of this contamination in museum collections . This paper reports on the determination of arsenic, mercury, and several organic pesticides on 17 objects that were recently repatriated to the Hupa tribe in northern California . Four samples were taken from each object: two for arsenic and mercury analysis via flame atomic absorption spectrophotometry and two for organic pesticide analysis via gas chromatography/mass spectrometry . Percent levels (wt/wt) of mercury were detected on many samples, and 0.001 to 0.183% (wt/wt) levels of p-dichlorobenzene, naphthalene, thymol, lindane, and/or DDT were detected on many of the samples . These results indicate that Hupa tribal members should not wear these objects in religious ceremonies, proper precautions should be followed when dealing with potentially contaminated objects, and that more serious consideration should be given to this issue at a national level.

Gig Sanit, 2003 Jan-Feb, (1), 32 - 5
{Soil cadmium pollution: environmental and hygienic aspects}; Mudryi IV; The paper deals with modes of detection of soil cadmium, with the effects of its pollution and with toxicity on man, microorganisms and soil biocenoses . It shows why cadmium migrates and translocates into plants . The problem of development of hygienic specifications of the metal is analyzed in relation to the protective properties of soil and the data obtained from the monitoring of a region.

J Hist Med Allied Sci, 2003 Jan, 58(1), 56 - 78
Transmission or recurrence? A historical dilemma of iatrogenic infections due to cytomegalovirus; Diosi P et al.; This article traces the changes in thought regarding the etiology of iatrogenic infections due to cytomegalovirus from the 1960s to the 1990s . Initial investigations using serologic and culture methods focused on how the virus was acquired . Following the application of molecular tools, theories on disease causality expanded beyond concerns of the microorganism itself to include aspects of the virus-host interaction and the host response . Eventually, the insights gained from the investigations into the etiology of iatrogenic cytomegalovirus disease were applied to other intracellular viruses . In addition, recognition of the factors responsible for reactivation of latent viruses had not only theoretical value, but also practical consequences.

Arthritis Rheum, 2003 Apr, 48(4), 927 - 34
Anakinra, a recombinant human interleukin-1 receptor antagonist (r-metHuIL-1ra), in patients with rheumatoid arthritis: A large, international, multicenter, placebo-controlled trial; Fleischmann RM et al.; OBJECTIVE: To evaluate the safety of anakinra (a recombinant human interleukin-1 receptor antagonist) in a large population of patients with rheumatoid arthritis (RA), typical of those seen in clinical practice . METHODS: A total of 1,414 patients were randomly assigned to treatment with 100 mg of anakinra or placebo, administered daily by subcutaneous injection . Background medications included disease-modifying antirheumatic drugs, corticosteroids, and nonsteroidal antiinflammatory drugs, alone or in combination . The primary end point was safety, which was evaluated by adverse events (including infections), discontinuation from study due to adverse events, and death . RESULTS: Safety was evaluated in 1,399 patients (1,116 in the anakinra group and 283 in the placebo group; 15 patients were randomized but did not receive any study drug) during the initial 6-month, double-blind, placebo-controlled phase of this long-term safety study . Baseline demographics, disease characteristics, and concomitant medications were similar between the 2 groups . The study group included patients with numerous comorbid conditions and a wide range of RA disease activity . Serious adverse events occurred at a similar rate in the anakinra group and the placebo group (7.7% and 7.8%, respectively) . Serious infectious episodes were observed more frequently in the anakinra group (2.1% versus 0.4% in the placebo group) . The rate of withdrawal due to adverse events was 13.4% in the anakinra group and 9.2% in the placebo group . CONCLUSION: Results from this large, placebo-controlled safety study demonstrate that anakinra is safe and well tolerated in a diverse population of patients with RA, including those with comorbid conditions and those using multiple combinations of concomitant therapies . Although the frequency of serious infection was slightly higher in the anakinra group, no infection was attributed to opportunistic microorganisms or resulted in death.

J Ind Microbiol Biotechnol, 2003 May, 30(5), 252 - 9 Epub 2003 Mar 25.
Enzymatic synthesis of chiral intermediates for pharmaceuticals; Patel R et al.; There has been an increasing awareness of the enormous potential of microorganisms and enzymes for the transformation of synthetic chemicals with high chemo-, regio- and enatioselective manner . Chiral intermediates are in high demand by pharmaceutical industries for the preparation bulk drug substances . In this review article, microbial/enzymatic processes for the synthesis of chiral intermediates for antihypertensive drugs, melatonin receptor agonists, and beta3-receptor receptor agonists are described.

Russ J Immunol, 1999 Dec, 4(4), 303 - 305
Current Features of Secondary (Acquired) Types of Immune Deficiency; Kovalchuk LV et al.; Secondary (acquired) types of immune deficiencies (SID) take a leading place in practice of modern clinical immunology . The causes for SID development are extremely variable . Special attention is concerned with accumulating facts about target action of microorganisms, and first of all viruses, on certain processes in immune system . Damageable action of HIV-1 on cell elements expressing CD4 molecules is known in most precise manner . It is noteworthy that the search of real molecular defects, induced by microorganisms in immune system is required . It is not to be ruled out that the increased level of apoptosis of immune system cells is one of the causes of SID . The basis of it is disbalance between positive and negative activation processes of immunocompetent cells . Multiple factors may serve as apoptogens, including drugs (glucocorticoids etc.), xenobiotics, physical factors (radiation) and many others . In practice of clinical laboratories a certain spectrum of immunological investigations is recommended that allows to diagnose the degree of immunopathology . At present, in clinical practice these methods are focused around flow cytometry (immunophenotyping), immunodiffusion and immunoenzyme tests (determination of immunoglobulins, cytokines, other soluble components of immune system), tests of estimation of immunocompetent cell activation, proliferation and differentiation . As a prospective, some methods, based on identification of molecular defects in cells and soluble factors of immune system, may be taken into consideration.

Cryobiology, 2003 Apr, 46(2), 146 - 52
Microbial contamination of embryos and semen during long term banking in liquid nitrogen; Bielanski A et al.; We report on microbial contamination of embryos and semen cryopreserved in sealed plastic straws and stored for 6-35 years in liquid nitrogen . There were 32 bacterial and 1 fungal species identified from randomly drawn liquid nitrogen, frozen semen, and embryos samples stored in 8 commercial and 8 research facility liquid nitrogen (LN) tanks . The identified bacteria represented commensal or environmental microorganisms and some, such as Escherichia coli, were potential or opportunistic pathogens for humans and animals . Stenotrophomonas maltophilia was the most common contaminant identified from the samples and was further shown to significantly suppress fertilization and embryonic development in vitro . Analysis of the strains by pulsed field gel electrophoresis revealed restriction patterns with no relatedness indicating that there was no apparent cross-contamination of S . maltophilia strains between the germplasm and liquid nitrogen samples . In addition, no transmission of bovine viral diarrhea virus (BVDV) and bovine herpesvirus-1 (BHV-1) from infected semen and embryos straws to clean germplasm stored in the same LN tanks or LN was detected.

Pharmazie, 2003 Mar, 58(3), 204 - 10
Effects of medicinal compounds on the differentiation of the eukaryotic microorganism dictyostelium discoideum: can this model be used as a screening test for reproductive toxicity in humans?
Dannat K, Tillner J, Winckler T, Weiss M, Eger K, Dingermann T.
Dictyostelium discoideum is a single-cell, eukaryotic microorganism that can undergo multicellular development in order to produce dormant spores . We investigated the capacity of D . discoideum to be used as a rapid screening system for potential developmental toxicity of compounds under development as pharmaceuticals . We used a set of four transgenic D . discoideum strains that expressed a reporter gene under the control of promoters that are active at certain time periods and in distinct cell types during D . discoideum development . We found that teratogens such as valproic acid, tretinoin, or thalidomide interfered to various extents with D . discoideum development, and had different effects on prestalk and prespore cell-specific reporter gene expression . Phenytoin was inactive in this assay, which may point to limitations in metabolization of the compound in Dictyostelium required to exert developmental toxicity . D . discoideum cell culture is cheap and easy to handle compared to mammalian cell cultures or animal teratogenicity models . Although the Dictyostelium-based assay described in this report may not securely predict the teratogenic potential of these drugs in humans, this organism may be qualified for rapid large-scale screenings of synthetic compounds under development as new pharmaceuticals for their potential to interfere with developmental processes and thus help to reduce the amount of teratogenicity tests in animal models.

Environ Toxicol Chem, 2003 Apr, 22(4), 722 - 9
Mineralization of aged atrazine, terbuthylazine, 2,4-D, and mecoprop in soil and aquifer sediment; Johannesen H et al.; The effect of aging of the herbicides atrazine, terbuthylazine, 2,4-D, and mecoprop on their bioavailability to degrading microorganisms was studied in soil and aquifer sediment . 14C-ring-labeled herbicide (2.5 mg/kg) was added to sterilized soil or aquifer sediment and stored at 10 degrees C for up to 103 d before inoculation with either the atrazine and terbuthylazine-degrading Pseudomonas sp . strain ADP (atrazine-degrading Pseudomonas) or an enriched culture able to mineralize 2,4-D and mecoprop . The initial mineralization rate and recovery of 14CO2 after 62 to 113 d of incubation were used as measures of the availability of the compounds to the microorganisms . Aging in soil reduced the initial mineralization of atrazine . Thus, only 17% of the added 14C-atrazine had been mineralized after 21 h of incubation when aged for 88 d as compared with 33% when the atrazine had been aged for 1 d . 14CO2 recovery was only 58% after 88 d of aging as compared with 81% when aged for 1 d . A similar effect of aging was seen with terbuthylazine . With 2,4-D, the effect of aging in soil on mineralization by the enriched culture was much smaller . Aging had no effect on mineralization of mecoprop in soil or on mineralization of any of the herbicides in aquifer sediment.

Environ Toxicol Chem, 2003 Apr, 22(4), 712 - 7
Dynamics of organochlorine pesticides in soils from a southeastern region of Argentina; Miglioranza KS et al.; Monitoring of organochlorine pesticides (OCPs) was carried out to identify and quantify the contribution of point and nonpoint sources to the total OCP flux in a southeastern region of Argentina . Natural, recreational, and agricultural soils located in the surrounding of a lagoon were analyzed by gas chromatography with electron-capture detector . Physical and chemical characteristics (texture, humidity, and organic matter content) were determined at different depths (0-15 cm, 15-30 cm, and 45-55 cm) . The pattern of OCP distribution was similar in all soil sampled, with DDT and metabolites > hexachlorocyclohexanes (HCHs) > heptachlor > chlordanes . The highest values of OCPs (656.1 ng/g dry wt) were found in the surface natural soil despite its never having received direct OCP application . This would be mainly due to the high organic matter content of the surface natural soils as well as its topographic position (highland hills), with main winds arriving from agricultural areas . Microorganism abundance and edaphic biota in the upper layer would justify the high levels of metabolites found . The agricultural soil (intensive tillage) also showed the highest OCP values (30.19 ng/g dry wt) in the surface horizon . Because of management practices, volatilization could have been one of the major causes of pesticide loss from this target area . Recreational soil showed the lowest OCP levels in the surface layer because of weathering that occurs when the nearby lagoon floods this zone . Our results show that, although most of these pesticides are banned, they are present in these soils and the atmospheric transport and deposition would be the major processes for distributing OCPs from target to natural areas.

Ying Yong Sheng Tai Xue Bao, 2002 Dec, 13(12), 1713 - 5
{Diversity of the microorganisms degrading aromatic hydrocarbons}; Qiu J et al.; Aromatic hydrocarbons are biological xenobiotics . Indigenous microorganism groups turn from being unfamiliar with into active response to environmental changes by their adaptability to environment . There are changes in their genetic background, and then, the biodiversity turns into being . This paper reviewed the microorganism groups resources, biocharacters, genetic background, evolvement and their adaptability to environment; described the genetic information, expression and regulatory for some species in detail; and pointed out that the degradation of aromatic hydrocarbons depended mainly on new microorganisms formed by genetic engineering and on their highly efficient metabolic regulatory.

Ying Yong Sheng Tai Xue Bao, 2002 Dec, 13(12), 1575 - 8
{Effect of combined application of bioorganic manure and inorganic nitrogen fertilizer on soil nitrogen supplying characteristics}; Zhang Y et al.; Pot experiment was carried out to study the effect of application bioorganic manure on soil nitrogen supplying characteristics . Compared with the application of raw rice straw, a combined application of bio-organic manure and inorganic nitrogen fertilizer could boost the content of microbial biomass carbon and nitrogen, and the increased percentage could be 300-400% . A strong immobilization of nitrogen by microorganisms was always followed by a net N mineralization, which was mostly favorable for the growth and development of plant, and improved the use efficiency of nitrogen fertilizer.

Curr Med Chem Anti-Canc Agents, 2002 Jul, 2(4), 485 - 537
Cytotoxic anticancer candidates from natural resources; Kim J et al.; Natural products have been regarded as important sources that could produce potential chemotherapeutic agents . Over 50% of anticancer drugs approved by United States Food and Drug Administration since 1960 were originated from the natural resources, especially from terrestrial plants . Based on cytotoxicity bioassay, over 400 compounds have been isolated from plants, marine organisms and microorganisms from the period of 1996 to 2000 . Recently, interest of natural product research has slowly moved to marine organisms . As a result, almost 50% of reported cytotoxic compounds were isolated from marine organisms such as sponges and corals . Also, traditional cytotoxic compounds of acetogenins, alkaloids and terpene skeletons have been reported continuously . In this review, we will present the cytotoxic compounds obtained from natural sources from 1996 to 2000, and the structures and cytotoxic activity of natural compounds isolated from territorial, marine and microorganism resources.

Arch Microbiol, 2003 Apr, 179(4), 258 - 65 Epub 2003 Feb 28.
Characterization of a new keratinolytic bacterium that completely degrades native feather keratin; Riffel A et al.; A novel feather-degrading microorganism was isolated from poultry waste, producing a high keratinolytic activity when cultured on broth containing native feather . Complete feather degradation was achieved during cultivation . The bacterium presents potential use for biotechnological processes involving keratin hydrolysis . Chryseobacterium sp . strain kr6 was identified based on morphological and biochemical tests and 16S rRNA sequencing . The bacterium presented optimum growth at pH 8.0 and 30 degrees C; under these conditions, maximum feather-degrading activity was also achieved . Maximum keratinase production was reached at 25 degrees C, while concentration of soluble protein was similar at both 25 and 30 degrees C . Reduction of disulfide bridges was also observed, increasing with cultivation time . The keratinase of strain kr6 was active on azokeratin and azocasein as substrates, and presented optimum pH and temperature of 7.5 and 55 degrees C, respectively . The keratinase activity was inhibited by 1,10-phenanthroline, EDTA, Hg(2+), and Cu(2+) and stimulated by Ca(2+).

Biotechnol Prog, 2003 Mar-Apr, 19(2), 495 - 500
Purification and properties of a NADPH-dependent erythrose reductase from the newly isolated Torula corallina; Lee JK et al.; Torula corallina (KCCM-10171) is a yeast strain that is currently used for the industrial production of erythritol and has the highest erythritol yield ever reported for an erythritol-producing microorganism . Production of erythritol in T . corallina is catalyzed by erythrose reductase, an enzyme that converts erythrose to erythritol using NADPH as a cofactor . In this study, NADPH-dependent erythrose reductase was purified to homogeneity from the newly isolated T . corallina . The relative molecular weight of the erythrose reductase as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size exclusion chromatography was 35.4 and 71.0 kDa, respectively, indicating that the enzyme is dimeric . This enzyme catalyzed both erythrose reduction and erythritol oxidation; both enzyme activities required NADP(H) . The pH and temperature optima for erythrose reduction and erythritol oxidation were 6.0, 40 degrees C and 8.0, 45 degrees C, respectively . The sequence of the first 10 amino acids of this enzyme was N-V-K-N-F-Y-Q-P-N-D . The affinity (K(m)( )()= 7.12 mM) of the enzyme for erythrose was comparable to that of other known erythrose reductases, and the specificity for erythrose was very high, resulting in no production of other polyols, which may explain the high erythritol yield observed in this strain.

Diabetes Metab Res Rev, 2003 Mar-Apr, 19(2), 89 - 100
A role for innate immunity in type 1 diabetes?
Beyan H, Buckley LR, Yousaf N, Londei M, Leslie RD.
Two arms of the immune system, innate and adaptive immunity, differ in their mode of immune recognition . The innate immune system recognizes a few highly conserved structures on a broad range of microorganisms . On the other hand, recognition of self or autoreactivity is generally confined to the adaptive immune response . Whilst autoimmune features are relatively common, they should be distinguished from autoimmune disease that is infrequent . Type 1 diabetes is an immune-mediated disease due to the destruction of insulin secreting cells mediated by aggressive immune responses, including activation of the adaptive immune system following genetic and environmental interaction . Hypotheses for the cause of the immune dysfunction leading to type 1 diabetes include self-reactive T-cell clones that (1) escape deletion in the thymus, (2) escape from peripheral tolerance or (3) escape from homeostatic control with an alteration in the immune balance leading to autoimmunity . Evidence, outlined in this review, raises the possibility that changes in the innate immune system could lead to autoimmunity, by either priming or promoting aggressive adaptive immune responses . Hostile microorganisms are identified by genetically determined surface receptors on innate effector cells, thereby promoting clearance of these invaders . These innate effectors include a few relatively inflexible cell populations such as monocytes/macrophages, dendritic cells (DC), natural killer (NK) cells, natural killer T (NKT) cells and gammadelta T cells . Recent studies have identified abnormalities in some of these cells both in patients with type 1 diabetes and in those at risk of the disease . However, it remains unclear whether these abnormalities in innate effector cells predispose to autoimmune disease . If they were to do so, then modulation of the innate immune system could be of therapeutic value in preventing immune-mediated diseases such as type 1 diabetes .

Biotechnol Bioeng, 2003 Jun 20, 82(6), 715 - 24
Influence of morphology on the near-infrared spectra of mycelial biomass and its implications in bioprocess monitoring; Vaidyanathan S et al.; Bioprocesses that employ mycelial microorganisms are commercially important . The application of optical techniques for the measurement of biomass in such processes is limited by the morphological heterogeneity exhibited by the mycelial microorganism employed . We investigated the influence of morphology on the near-infrared (NIR) spectra of the biomass of Streptomyces fradiae, a filamentous microorganism, by studying the spectra of mycelial suspensions that were manipulated to generate a range of morphological forms . Computerized image analysis was used to characterize the morphological forms . Principal component analysis was used to assess the spectral variations and study correlations to the manipulated mycelial morphology . Although morphology was found to influence the near infrared transmittance spectra of biomass, the influence was less pronounced than in the visible region, the spectral information at longer wavelengths (1600-2350 nm) showing greater stability to morphological variations . Long-wave NIR spectral information is therefore likely to be more useful in estimating biomass in mycelial bioprocesses . Furthermore, the NIR reflectance spectra of dried biomass were found to show correlations to the morphological variations introduced, suggesting that NIR spectra may be useful in obtaining morphology related information .

Biotechnol Bioeng, 2003 Jun 20, 82(6), 627 - 39
Generally applicable fed-batch culture concept based on the detection of metabolic state by on-line balancing; Jobe AM et al.; In many microorganisms, flux limitations in oxidative metabolism lead to the formation of overflow metabolites even under fully aerobic conditions . This can be avoided if the specific growth rate is controlled at a low enough value . This is usually accomplished by controlling the substrate feeding profile in a fed-batch process . The present work proposes a control concept which is based on the on-line detection of metabolic state by on-line calculation of mass and elemental balances . The advantages of this method are: 1) the check of measurement consistency based on all of the available measurements, 2) the minimum requirement of a priori knowledge of metabolism, and 3) the exclusive use of simple and established on-line techniques which do not require direct measurement of the metabolite in question . The control concept has been linked to a simple adaptive controller and applied to fed-batch cultures of S . cerevisiae and E . coli, organisms which express different overflow metabolites, ethanol and acetic acid, respectively . Oxidative and oxidoreductive states of S . cerevisiae and E . coli cultures were detected with high precision . As demonstrated by the formation of acetic acid in E . coli cultures, metabolic states could be correctly distinguished for systems for which traditional methods, such as respiratory quotient (RQ), are insensitive . Hence, it could be shown that the control concept allowed avoidance of overflow metabolite formation and operation at maximum oxidative biomass productivity and oxidative conversion of substrate into biomass . Based on mass and elemental balances, the proposed method additionally provides a richness of additional information, such as yield coefficients and estimation of concentrations and specific conversion rates . These data certainly help the operator to additionally evaluate the state of the process on-line .

Gastric Cancer, 2003, 6(1), 60 - 3
Malignant lymphoma occurring in the residual stomach following gastrectomy: plus discussion based on the literature in Japan; Oshita H et al.; Malignant lymphoma of the remnant stomach was diagnosed in a 53-year-old man 8 years after gastrectomy for a perforated gastric ulcer . Endoscopic examination demonstrated protruding lesions spreading over the entire residual stomach, and biopsy revealed malignant lymphoma . Rectal cancer was diagnosed simultaneously . The residual stomach was completely excised, with splenectomy, in parallel with low anterior resection of the rectum . Histological studies revealed that the lesion in the residual stomach was a lymphoma of the diffuse, large-cell type, according to the Lymphoma-Leukemia Study Group (LSG) classification, with positivity for CD20 and CD45RA, leading to a diagnosis of B-cell lymphoma . Helicobacter pylori microorganisms were found on the luminal surface of the tumor . Despite postoperative chemotherapy, the patient died of disseminated lymphoma 34 months later . Although malignant lymphoma occurring in the residual stomach following gastrectomy is rare, particular attention should be given to the possible presence of a malignant tumor when examining the residual stomach following gastrectomy.

J Mol Microbiol Biotechnol, 2003, 5(1), 46 - 56
Metagenomes of complex microbial consortia derived from different soils as sources for novel genes conferring formation of carbonyls from short-chain polyols on Escherichia coli; Knietsch A et al.; Metagenomic DNA libraries from three different soil samples (meadow, sugar beet field, cropland) were constructed . The three unamplified libraries comprised approximately 1267000 independent clones and harbored approximately 4.05 Gbp of environmental DNA . Approximately 300000 recombinant Escherichia coli strains of each library per test substrate were screened for the production of carbonyls from short-chain (C2 to C4) polyols such as 1,2-ethanediol, 2,3-butanediol, and a mixture of glycerol and 1,2-propanediol on indicator agar . Twenty-four positive E . COLI clones were obtained during the initial screen . Fifteen of them contained recombinant plasmids, designated pAK201-215, which conferred a stable carbonyl-forming phenotype on E . coli Sequencing revealed that the inserts of pAK201-215 encoded 26 complete and 14 incomplete predicted protein-encoding genes . Most of these genes were similar to genes with unknown functions from other microorganisms or unrelated to any other known gene . The further analysis was focused on the 7 plasmids (pAK204, pAK206, pAK208, and pAK210-213) recovered from the positive clones, which exhibited an NAD(H)-dependent alcohol oxidoreductase activity with polyols or the correlating carbonyls as substrates in crude extracts . Three genes (ORF6, ORF24, and ORF25) conferring this activity were identified during subcloning of the inserts of pAK204, pAK211, and pAK212 . The sequences of the three deduced gene products revealed no significant similarities to known alcohol oxidoreductases, but contained putative glycine-rich regions, which are characteristic for binding of nicotinamide cofactors .

Int J Food Microbiol, 2003 May 25, 83(1), 39 - 48
Standardization of diagnostic PCR for the detection of foodborne pathogens; Malorny B et al.; In vitro amplification of nucleic acids using the polymerase chain reaction (PCR) has become, since its discovery in the 1980s, a powerful diagnostic tool for the analysis of microbial infections as well as for the analysis of microorganisms in food samples . However, despite its potential, PCR has neither gained wide acceptance in routine diagnostics nor been widely incorporated in standardized methods . Lack of validation and standard protocols, as well as variable quality of reagents and equipment, influence the efficient dissemination of PCR methodology from expert research laboratories to end-user laboratories . Moreover, the food industry understandably requires and expects officially approved standards . Recognizing this, in 1999, the European Commission approved the research project, FOOD-PCR , which aims to validate and standardize the use of diagnostic PCR for the detection of pathogenic bacteria in foods . The present review focuses on the harmonization procedure and standardization criteria for detection of foodborne pathogens by PCR . The progress of standardization so far and future perspectives of diagnostic PCR are discussed.

Nat Rev Cancer, 2003 Apr, 3(4), 276 - 85
Radical causes of cancer; Hussain SP et al.; Free radicals are ubiquitous in our body and are generated by normal physiological processes, including aerobic metabolism and inflammatory responses, to eliminate invading pathogenic microorganisms . Because free radicals can also inflict cellular damage, several defences have evolved both to protect our cells from radicals--such as antioxidant scavengers and enzymes--and to repair DNA damage . Understanding the association between chronic inflammation and cancer provides insights into the molecular mechanisms involved . In particular, we highlight the interaction between nitric oxide and p53 as a crucial pathway in inflammatory-mediated carcinogenesis.

J Int Acad Periodontol, 2002 Oct, 4(4), 119 - 25
Neutrophil-mediated host response to Porphyromonas gingivalis; Kantarci A et al.; Periodontal diseases are infections initiated by specific species of microorganisms and are among the most common human infections . The pathogenesis of periodontitis is mediated by interactions between host and microbial factors, complicated by genetic and environmental risk factors . Periodontal disease also represents a unique model in which to study the roles of bacterial and host-related factors, a model in which patients do not suffer from life-threatening disease . The aim of this paper is to focus on recent findings relating to neutrophil-mediated host response mechanisms in Porphyromonas gingivalis-induced periodontal disease . Virulence factors of Porphyromonas gingivalis such as the gingipains, fimbrillin peptides, capsule polysaccharides, lipopolysaccharides, haemagglutinating and haemolysing activities, toxic products of metabolism, outer membrane vesicles, and other enzymes have important roles in eliciting host responses in various ways . These factors significantly affect epithelial/endothelial cells, but their major effect is observed on the modulation of neutrophil response . Periodontitis represents an important model for neutrophil-mediated host tissue injury . In this model, neutrophils, primed or stimulated by the presence or persistence of infection, express an elevated and excessive response . This, in turn, leads to tissue destruction mediated by neutrophil activity . It is essential to understand the mechanisms underlying the interactions between the neutrophils and the microbial virulence factors to be able to develop rational, novel treatment strategies.

Bull Acad Natl Med, 2002, 186(8), 1377 - 88; discussion 1388-9
{GMOs in food: risk assessment and management; scientific and regulatory characteristics}; Casse F et al.; Genetic transformation constitutes a new tool for improvement of microorganisms, animals and plants used in food . Foreseeable risks are evoked, as well as management measures to avoid GMO unsuspected risks . Few risks are specific to GMOs . Present elements of french and european regulations concerning placing on the market and follow up GMOs and other novel foods are described.

Ann Pharm Fr, 2003 Mar, 61(2), 87 - 95
{GMOs in food: risk assessment, scientific management and regulatory aspects}; Casse F et al.; Genetic transformation constitutes a new tool for improvement of microorganisms, animals and plants used in food . We present foreseeable risks, as well as management measures to avoid unsuspected risks of GMOs . Few risks are specific to GMOs . Present elements of French and European regulations concerning placing on the market and follow up GMOs and other novel foods are described.

Cardiovasc Toxicol, 2003, 3(1), 43 - 70
Inhibitors of the complement system currently in development for cardiovascular disease; Pugsley MK et al.; Controlled activation of the complement system is critical to the host-defense response of the immune system . Activated complement is responsible for the stimulation of a localized protective inflammatory response to either invading microorganisms or foreign molecules (toxins) . However, the autologous activation of the complement system can have devastating consequences on many organ systems . This review discusses the various pathways involved in the activation of the complement system and the multiple levels of control established within the body to regulate activation . It also focuses on the role of complement activation in cardiovascular disease, especially myocardial ischemia and reperfusion injury as well as in cardiopulmonary bypass procedures . Lastly, this review also provides a comprehensive overview of both biologically derived proteins and chemically developed inhibitors of the complement system that range from those that are currently in the discovery stage to those that are in clinical development as novel therapeutic agents.

Virology, 2003 Mar 15, 307(2), 406 - 13
Neutralizing antibodies against conserved domains of p15E of porcine endogenous retroviruses: basis for a vaccine for xenotransplantation?
Fiebig U, Stephan O, Kurth R, Denner J.
Porcine xenotransplants may offer a potential solution to the problem posed by the limited supply of allotransplants . However, xenotransplantation may be associated with the risk of transmission of microorganisms, in particular of porcine endogenous retroviruses (PERVs) that are an integral part of the porcine genome and able to infect human cells in vitro . Possible strategies to prevent virus transmission include the development of PERV knockout animals or of effective vaccines . When antisera prepared against the main structural proteins of PERV were screened, a goat antiserum against the recombinant ectodomain of the transmembrane envelope protein p15E was found to neutralize PERV infectivity . Epitope mapping using overlapping peptides revealed two epitopes, E1 (GPQQLEK) and E2 (FEGWFN) . These sequences are identical for all PERVs and are highly conserved among all gammaretroviruses . Interestingly, antibodies isolated from AIDS patients and specific for sequences of HIV-1 partially homologous with E2 (Mab4E10, LWNWFN) or located in close proximity to E2 (Mab2F5, ELDKWA) are known to neutralize several strains of HIV-1 . It is the first report showing epitope mapping of gammaretrovirus-specific neutralizing antibodies and demonstrating similarity to corresponding epitopes in HIV . These domains of the transmembrane proteins of different retroviruses are an effective target for neutralizing antibodies and may be a useful antigen to create an antiretroviral vaccine.

J Vet Med B Infect Dis Vet Public Health, 2003 Apr, 50(3), 118 - 20
Aetiology of ovine footrot in the Portuguese region of Alto Alentejo; Jimenez R et al.; In this work, we found it appropriate to carry out a study directed towards isolating and identifying the entailed microorganisms which trigger off footrot in sheep, placing special emphasis on the serotipification of the different Dichelobacter nodosus species . With this goal in mind four flocks from the Portuguese region of 'Alto Alentejo' were selected, all of them had one common feature: their main health problem was ovine footrot . We also set out to determine the elastolitic capacity of isolated strict-anaerobic bacteria, in order to be able to clarify the direct involvement of these microorganisms in the outbreak of this infectious process.

Biochemistry, 2003 Apr 8, 42(13), 3766 - 76
The high-resolution structure of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase reveals a twist in the plane of bound phosphoenolpyruvate; Shumilin IA et al.; 3-Deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHPS), the first enzyme of the aromatic biosynthetic pathway in microorganisms and plants, catalyzes the aldol-like condensation of phosphoenolpyruvate (PEP) and D-erythrose 4-phosphate (E4P) with the formation of DAHP . The native and the selenomethionine-substituted forms of the phenylalanine-regulated isozyme {DAHPS(Phe)} from Escherichia coli were crystallized in complex with PEP and a metal cofactor, Mn(2+), but the crystals displayed disorder in their unit cells, preventing satisfactory refinement . However, the crystal structure of the E24Q mutant form of DAHPS(Phe) in complex with PEP and Mn(2+) has been determined at 1.75 A resolution . Unlike the tetrameric wild-type enzyme, the E24Q enzyme is dimeric in solution, as a result of the mutational perturbation of four intersubunit salt bridges that are critical for tetramer formation . The protein chain conformation and subunit arrangement in the crystals of E24Q and wild-type DAHPS are very similar . However, the interaction of Mn(2+) and PEP in the enzymatically active E24Q mutant complex differs from the Pb(2+)-PEP and Mn(2+)-phosphoglycolate interactions in two enzymatically inactive wild-type complexes whose structures have been determined previously . The geometry of PEP bound in the active site of the E24Q enzyme deviates from planarity due to a 30 degrees twist of the carboxylate plane relative to the enol plane . In addition, seven water molecules are within contact distance of PEP, two of which are close enough to its C2 atom to serve as the nucleophile required in the reaction.

J Int Acad Periodontol, 2000 Jul, 2(3), 59 - 63
Understanding the pathogenesis of periodontitis: a century of discovery; Williams RC et al.; The twentieth century was an especially notable time for research advances into the understanding of the aetiology and the pathogenesis of periodontitis . Beginning in 1900, using dark-field preparations, investigators began to implicate certain microorganisms, such as amoebae, as causing periodontitis . Today, using modern molecular techniques, we believe we have identified the primary causative agents of adult periodontitis . In 1900, histological sections of periodontal tissues at autopsy provided clues as to how periodontal pockets probably formed . Today there is a wealth of cellular and molecular data that suggest the actual pathways that the susceptible host uses to initiate periodontal tissue destruction . We now also appreciate that periodontitis may be a significant risk factor for systemic disease . A very exceptional 'century of discovery' into the nature of periodontal diseases should now lead to a new era of better diagnosis, prevention and treatment for this ubiquitous disease.

Parasitology, 2003 Mar, 126(Pt 3), 195 - 202
Membrane-bound acid phosphatase (MAP) from Entamoeba histolytica has phosphotyrosine phosphatase activity and disrupts the actin cytoskeleton of host cells; Aguirre-Garcia MM et al.; Protein tyrosine phosphatases (PTPases) have been described as virulence factors in different pathogenic microorganisms . The pathogenic process by Enatamoeba histolytica is a multifactorial phenomenon that occurs in 3 steps: adhesion, cytolytic and cytotoxic effect, and phagocytosis . Lytic enzymes may participate during the second part of this process . In this work, we determined that purified membrane-bound acid phosphatase (MAP) from E . histolytica trophozoites has PTPase activity . The enzyme specifically dephosphorylated O-phospho-L-tyrosine at optimum pH of 5.0, with little activity towards O-phospho-L-serine, O-phospho-L-threonine, and ATP . It was inhibited by ammonium molybdate and sodium tungstate, and trifluoperazine did not show any effect . A monoclonal antibody against the catalytic domain of the human placental PTPase 1B, cross-reacted with a 55 kDa molecule present in the solubilized fraction . The interaction of the amoebic PTPase with HeLa cells resulted in the alteration of the cell actin cytoskeleton by disruption of the actin stress fibres.

Environ Technol, 2003 Feb, 24(2), 187 - 96
Hydrodynamic characterization of materials for gas treatment by biofluidization; Benesse M et al.; The fluidization of possible microorganism supports has been studied . The fluidization of scrap-wood particles and of polyurethane foam cylinders was characterized by measuring their minimum fluidization velocities and compared with predictions from the literature . Minimum fluidization velocity increases with the water content of the scrap-wood more than predicted by change in size and density, thus probably involving interparticle forces at high moisture levels . Two types of polyurethane foam cylinders have been also characterized from fixed bed to entrainment regimes . Measurement of the pressure drop in the fixed bed state and of the minimum fluidization velocity suggest that flow occurs through these porous particles, as well as around them . Nevertheless the contribution of the internal particle porosity to the flow decreases from the fixed bed to the entrained regime . The onset of entrainment is well predicted by correlations established for non-porous materials.

Mikrobiol Z, 2002 Nov-Dec, 64(6), 28 - 34
{Isolation from the environment of strains of microorganisms with glucose oxidase activity}; Smotrova NG et al.; Strains of microorganisms (13) producing enzyme glucoseoxidase were isolated from various samples of soil . The ability to oxidase glucose in presence of oxygen is most expressed in representatives of genera Aureobasidium, Aspergillus, Penicillium . Media containing the KJ-starch indicator were used at primary step for isolation of microorganisms from the environment . The strain, identified as Aureobasidium pullulans (de Bary) Arnaud, isolated from soil had the greatest glucoseoxidase activity, remained active after lyophilization, grew on the medium, which contained low concentration of glucose and nitrate for candides . Use of its cells as a glucoseoxidase sensor control allowed to detect glucose in the liquids with concentration from 0.027 mM/l.

Protoplasma, 2003 Mar, 220(3-4), 119 - 29
Proteolysis and cell death in clover leaves is induced by grazing; Kingston-Smith AH et al.; Programmed plant cell death is a widespread phenomenon resulting in the formation of xylem vessels, dissected leaf forms, and aerenchyma . We demonstrate here that some characteristics of programmed cell death can also be observed during the cellular response to biotic and abiotic stress when plant tissue is ingested by grazing ruminants . Furthermore, the onset and progression of plant cell death processes may influence the proteolytic rate in the rumen . This is important because rapid proteolysis of plant proteins in ruminants is a major cause of the inefficient conversion of plant to animal protein resulting in the release of environmental N pollutants . Although rumen proteolysis is widely believed to be mediated by proteases from rumen microorganisms, proteolysis and cell death occurred concurrently in clover leaves incubated in vitro under rumenlike conditions (maintained anaerobically at 39 degrees C) but in the absence of a rumen microbial population . Under rumenlike conditions, both red and white clover cells showed progressive loss of DNA, but this was only associated with fragmentation in white clover . Cell death was indicated by increased ionic leakage and the appearance of terminal deoxynucleotidyl transferase-mediated dUTP-nick-end-labelled nuclei . Foliar protein decreased to 50% of the initial values after 3 h incubation in white clover and after 4 h in red clover, while no decrease was observed in ambient (25 degrees C, aerobic) incubations . In white clover, decreased foliar protein coincided with an increased number of protease isoforms.

Appl Microbiol Biotechnol, 2003 Feb, 60(6), 743 - 7 Epub 2003 Jan 30.
Achievement of rapid osmotic dehydration at specific temperatures could maintain high Saccharomyces cerevisiae viability; Laroche C et al.; Various methods have been tried to prevent cell mortality during dehydration, but the reasons why microorganisms die when submitted to dehydration and rehydration are not well understood . The aim of this study was to further investigate the reasons for yeast mortality during dehydration . Osmotic dehydration and rehydration of Saccharomyces cerevisiae W303-1A were performed at different temperatures . Two different approaches were used: isothermic treatments (dehydration and rehydration at the same temperature), and cyclic treatments (dehydration at an experimental temperature and rehydration at 25 degrees C), with significant differences in viability found between the different treatments . Dehydration at lower and higher temperatures gave higher viability results . These experiments allowed us to propose a hypothesis that relates mortality to a high water flow through an unstable membrane during phase transition.

Physiol Rev, 2003 Apr, 83(2), 337 - 76
Clinical, cellular, and molecular aspects of cancer invasion; Mareel M et al.; Invasion causes cancer malignancy . We review recent data about cellular and molecular mechanisms of invasion, focusing on cross-talk between the invaders and the host . Cancer disturbs these cellular activities that maintain multicellular organisms, namely, growth, differentiation, apoptosis, and tissue integrity . Multiple alterations in the genome of cancer cells underlie tumor development . These genetic alterations occur in varying orders; many of them concomitantly influence invasion as well as the other cancer-related cellular activities . Examples discussed are genes encoding elements of the cadherin/catenin complex, the nonreceptor tyrosine kinase Src, the receptor tyrosine kinases c-Met and FGFR, the small GTPase Ras, and the dual phosphatase PTEN . In microorganisms, invasion genes belong to the class of virulence genes . There are numerous clinical and experimental observations showing that invasion results from the cross-talk between cancer cells and host cells, comprising myofibroblasts, endothelial cells, and leukocytes, all of which are themselves invasive . In bone metastases, host osteoclasts serve as targets for therapy . The molecular analysis of invasion-associated cellular activities, namely, homotypic and heterotypic cell-cell adhesion, cell-matrix interactions and ectopic survival, migration, and proteolysis, reveal branching signal transduction pathways with extensive networks between individual pathways . Cellular responses to invasion-stimulatory molecules such as scatter factor, chemokines, leptin, trefoil factors, and bile acids or inhibitory factors such as platelet activating factor and thrombin depend on activation of trimeric G proteins, phosphoinositide 3-kinase, and the Rac and Rho family of small GTPases . The role of proteolysis in invasion is not limited to breakdown of extracellular matrix but also causes cleavage of proinvasive fragments from cell surface glycoproteins.

Int Arch Allergy Immunol, 2003 Mar, 130(3), 180 - 92
Toll-like receptors and their function in innate and adaptive immunity; Heine H et al.; Over the past 3 years our knowledge about how we sense the microbial world has been fundamentally changed . It has been known for decades that microbial products, such as lipopolysaccharide, lipoproteins, or peptidoglycan, have a profound activity on human cells . Whereas the structure of many different pathogenic microbial compounds has been extensively studied and characterized, the molecular basis of their recognition by the cells of the innate immune system remained elusive for a long time . It was Charles Janeway {Cold Spring Harb Symp Quant Biol 1989;54/1:1-13} who developed the idea of microbial structures forming pathogen-associated molecular patterns that would be recognized by pattern recognition receptors . The discovery of the family of Toll receptors in species as diverse as DROSOPHILA and humans, and the recognition of their role in distinguishing molecular patterns that are common to microorganisms have led to a renewed appreciation of the innate immune system . Moreover, it is now clear that the activation of the innate immune system through mammalian Toll-like receptors has also an instructive role for the responses of the adaptive immune response and, thus, may influence allergic diseases such as asthma .

BMC Immunol . 2003 Mar 04;4(1):3.
Modulation of neutrophil function by the tripeptide feG; Mathison RD et al.; BACKGROUND: Neutrophils are critical in the defense against potentially harmful microorganisms, but their excessive and inappropriate activation can contribute significantly to tissue damage and a worsening pathology . Through the release of endocrine factors submandibular glands contribute to achieving a balance in neutrophil function by modulating the state of activation and migratory potential of circulating neutrophils . A putative hormonal candidate for these effects on neutrophils was identified as a heptapeptide named submandibular gland peptide T (SGP-T; sequence = TDIFEGG) . Since the tripeptide FEG, derived from SGP-T, and its D-amino acid analogue feG had similar inhibitory effects on inflammatory reactions, we investigated the effects of feG on human and rat neutrophil function . RESULTS: With human neutrophils feG had no discernible effect on oxidative burst or phagocytosis, but in picomolar amounts it reduced PAF-induced neutrophil movement and adhesion, and the binding of CD11b by 34% and that of CD16b close to control values . In the rat feG (10-11M) reduced the binding of CD11b and CD16 antibodies to PAF-stimulated circulating neutrophils by 35% and 43%, respectively, and at 100 micrograms/kilograms intraperitoneally feG reduced neutrophil in vivo migration by 40% . With ovalbumin-sensitized rats that were challenged with antigen, feG inhibited binding of antibodies against CD16b but not CD11b, on peritoneal leukocytes . CONCLUSIONS: The inhibitory effect of feG on neutrophil movement may be mediated by alterations in the co-stimulatory molecules CD11b and CD16.

Photochem Photobiol Sci, 2003 Jan, 2(1), 51 - 61
Interactive effects of ozone depletion and climate change on biogeochemical cycles; Zepp RG et al.; The effects of ozone depiction on global biogeochemical cycles, via increased UV-B radiation at the Earth's surface, have continued to be documented over the past 4 years . In this report we also document various effects of UV-B that interact with global climate change because the detailed interactions between ozone depletion and climate change are central to the prediction and evaluation of future Earth environmental conditions . There is increasing evidence that elevated UV-B has significant effects on the terrestrial biosphere with important implications for the cycling of carbon, nitrogen and other elements . Increased UV has been shown to induce carbon monoxide production from dead plant matter in terrestrial ecosystems, nitrogen oxide production from Arctic and Antarctic snowpacks, and halogenated substances from several terrestrial ecosystems . New studies on UV effects on the decomposition of dead leaf material confirm that these effects are complex and species-specific . Decomposition can be retarded, accelerated or remain unchanged . It has been difficult to relate effects of UV on decomposition rates to leaf litter chemistry, as this is very variable . However, new evidence shows UV effects on some fungi, bacterial communities and soil fauna that could play roles in decomposition and nutrient cycling . An important new result is that not only is nitrogen cycling in soils perturbed significantly by increased UV-B, but that these effects persist for over a decade . As nitrogen cycling is temperature dependent, this finding clearly links the impacts of ozone depletion to the ability of plants to use nitrogen in a warming global environment . There are many other potential interactions between UV and climate change impacts on terrestrial biogeochemical cycles that remain to be quantified . There is also new evidence that UV-B strongly influences aquatic carbon, nitrogen, sulfur, and metals cycling that affect a wide range of life processes . UV-B accelerates the decomposition of colored dissolved organic matter (CDOM) entering the sea via terrestrial runoff, thus having important effects on oceanic carbon cycle dynamics . Since UV-B influences the distribution of CDOM, there is an impact of UV-B on estimates of oceanic productivity based on remote sensing of ocean color . Thus, oceanic productivity estimates based on remote sensing require estimates of CDOM distributions . Recent research shows that UV-B transforms dissolved organic matter to dissolved inorganic carbon and nitrogen, including carbon dioxide and ammonium and to organic substances that are either more or less readily available to micro-organisms . The extent of these transformations is correlated with loss of UV absorbance by the organic matter . Changes in aquatic primary productivity and decomposition due to climate-related changes in circulation and nutrient supply, which occur concurrently with increased UV-B exposure, have synergistic influences on the penetration of light into aquatic ecosystems . New research has confirmed that UV affects the biological availability of iron, copper and other trace metals in aquatic environments thus potentially affecting the growth of phytoplankton and other microorganisms that are involved in carbon and nitrogen cycling . There are several instances where UV-B modifies the air sea exchange of trace gases that in turn alter atmospheric chemistry, including the carbon cycle.

Photochem Photobiol Sci, 2002 Jul, 1(7), 459 - 67
Photoreception and photomovements of microorganisms; Sgarbossa A et al.; Many freely motile microorganisms can perceive and transduce external photic stimuli to the motor apparatus, eventually moving, by means of various behavioural strategies, into environments in which the illumination conditions are the most favourable for their life . In different microorganisms, a wide range of chromophores operate as light detectors, each of them set in a special molecular pocket that, in its turn, can be linked to another component of the transduction chain . The diverse photosensors are organized in special (and in many cases dedicated) photoreceptor units or subcellular organelles . The main molecular mechanisms connecting the early event of photon absorption to the formation of the signalling state down to the dark steps of the transduction chain are discussed in a selected number of case examples . The possible importance of an intensive multidisciplinary approach to these problems in an evolutionary perspective is finally briefly outlined.

Bioorg Khim, 2003 Jan-Feb, 29(1), 3 - 20
{Proline-specific endopeptidases}; Besedin DV et al.; Prolyl endopeptidases, or post-proline-cleaving enzymes, are the specific endopeptidases that hydrolyze peptide substrates at the carbonyl of the internal Pro residue . All the currently known prolyl endopeptidases from animals, microorganisms, fungi, and plants as well as the post-proline-cleaving enzymes that do not exhibit the strict specificity to Pro are reviewed . The data on their physicochemical and catalytic properties, substrate specificity, inhibitors, sequences, and three-dimensional structures are discussed.

Nat Rev Immunol, 2003 Mar, 3(3), 253 - 7
Natural versus adaptive regulatory T cells; Bluestone JA et al.; The regulation of immune responses to self-antigens is a complex process that involves maintaining self-tolerance while retaining the capacity to mount robust immune responses against invading microorganisms . Over the past few years, many new insights into this process have been gained, leading to the re-emergence of the idea that regulatory T (T(Reg)) cells are a central mechanism of immune regulation . These insights have raised fundamental questions concerning what constitutes a T(Reg) cell, where they develop and what signals maintain T(Reg)-cell populations in a functional state . Here, we propose the existence of two subsets of CD4+ T(Reg) cells--natural and adaptive--that differ in terms of their development, specificity, mechanism of action and dependence on T-cell receptor and co-stimulatory signalling.

Int Endod J, 2003 Jan, 36(1), 20 - 6
Oral treponemes in primary root canal infections as detected by nested PCR; Rocas IN et al.; AIM: To investigate the prevalences of four Treponema species in primary root canal infections using a nested PCR assay . METHODOLOGY: Samples were obtained from 32 infected root canals . Twenty-two cases showed chronic asymptomatic periradicular lesions and 10 symptomatic cases were diagnosed as acute apical periodontitis . DNA extracted from the samples was initially amplified using universal 16S rDNA primers . A second round of amplification used the first PCR products to detect a specific fragment of the 16S rDNA of each Treponema denticola, T . socranskii, T . vincentii and T . pectinovorum . RESULTS: Bacteria were present in all cases sampled . T . denticola was detected in 77.3% of the asymptomatic cases, T . socranskii in 40.9%, T . vincentii in 18.2% and T . pectinovorum in 13.6% . In the cases diagnosed as acute apical periodontitis, T . denticola was detected in 80%, whilst T . socranskii and T . vincentii were detected in 40% and 10% of cases, respectively . No symptomatic case yielded T . pectinovorum . In general, nested PCR detected T . denticola in 78.1% of the cases, T . socranskii in 40.6%, T . vincentii in 15.6% and T . pectinovorum in 9.4% . At least one of the four Treponema species was found in 84.4% of the cases examined . CONCLUSIONS: The species T . denticola was detected in a large number of the cases examined: the prevalence of T . socranskii was also relatively high . The species T . vincentii and T . pectinovorum were also found, but in a smaller number of cases . Based on these data, the recognized pathogenicity of these microorganisms and their involvement with other oral diseases, they should be included in the restricted set of putative endodontic pathogens.

Ann Otol Rhinol Laryngol, 2003 Mar, 112(3), 197 - 201
Outcome of treatment with valacyclovir and prednisone in patients with Bell's palsy; Axelsson S et al.; Idiopathic facial paralysis, or Bell's palsy, shows a nonepidemic pattern that might indicate reactivation of a latent microorganism such as herpes simplex type I as a causative agent . Thirty percent of patients with Bell's palsy given no treatment will not recover completely, and 5% will have severe sequelae . The aim of this study was to find out whether treatment with an antiviral drug in combination with corticosteroids is more effective than no medical treatment at all in patients with Bell's palsy . Fifty-six consecutive adult patients attending the otorhinolaryngology department of the University Hospital of Lund from 1997 to 1999 were treated with 1 g of valacyclovir hydrochloride 3 times per day for 7 days and 50 mg of prednisone daily for 5 days, with the dose being reduced by 10 mg daily for the next 5 days . Fifty-six adult patients with Bell's palsy attending the same department between 1995 and 1996 who were given no medical treatment were studied retrospectively and used as the control group . Forty-nine patients (87.5%) in the treatment group recovered completely, as compared with 38 patients (68%) in the control group (p < .05) . One patient (1.8%) in the treatment group displayed severe sequelae, defined as a House-Brackmann score of IV or worse, as compared with 10 of 56 patients (18%) in the control group (p < .01) . Among patients over 60 years old, 10 of 10 in the treatment group had complete recovery, as compared with 5 of 12 patients in the control group (p < .01) . The present study showed a significantly better outcome in patients with Bell's palsy treated with valacyclovir and prednisone as compared with patients given no medical treatment . This difference in outcome was especially pronounced among elderly patients.

Appl Microbiol Biotechnol, 2003 Apr, 61(2), 118 - 22 Epub 2003 Jan 16.
Purification, cloning, sequencing and over-expression in Escherichia coli of a regioselective aliphatic nitrilase from Acidovorax facilis 72W; Chauhan S et al.; A regioselective aliphatic nitrilase from Acidovorax facilis 72W was purified and characterized, and the corresponding gene was cloned and sequenced . This nitrilase gene was over-expressed in Escherichia coli, generating a microorganism that efficiently and regioselectively catalyzes the conversion of aliphatic dinitriles to cyanocarboxylic acids . The high yields obtained, mild reaction conditions used, and robustness observed make this biocatalyst suitable for industrial applications.

Eur J Pediatr, 2003 Mar, 162(3), 122 - 8 Epub 2003 Jan 31.
Immunonutrients and neonates; Huang Y et al.; The gastrointestinal tract is the largest surface area of the body and the primary site for microorganisms, foreign antigens and toxins to gain entry to the host's internal milieu . The use of enteral feedings enriched with immune-enhancing ingredients is attracting considerable interest because there is increasing application of enteral feeding and appreciation of the role of the gut in the development of infection and of multiple organ failure in critically ill patients . CONCLUSION: in this review, we will discuss nutrients, such as glutamine, arginine, omega 3 fatty acids, nucleotides, probiotics, and lactoferrin, and how they might be used as immunonutrients in neonatal clinics.

Infect Immun, 2003 Apr, 71(4), 2095 - 101
Leishmania donovani-induced expression of suppressor of cytokine signaling 3 in human macrophages: a novel mechanism for intracellular parasite suppression of activation; Bertholet S et al.; Leishmania donovani protozoan parasites, the causative agent of visceral leishmaniasis, establish an infection partly by interfering with cytokine signaling in the host macrophages . Therefore, we investigated the expression of the suppressor of cytokine signaling (SOCS) genes in human macrophages infected with L . donovani . The expression of SOCS3 mRNA was induced transiently after exposure to live or heat-killed parasites, but not purified lipophosphoglycan, while that of other SOCS genes remained unchanged . SOCS3 gene expression was not dependent on phagocytosis or on cytokines released by L . donovani-infected macrophages, such as interleukin-1beta or tumor necrosis factor alpha . In addition, Leishmania used a different signaling pathway(s) than bacterial lipopolysaccharide to induce SOCS3 mRNA, as indicated by the kinetics of induction and sensitivity to polymyxin B inhibition . Finally, phosphorylation of the STAT1 transcription factor was significantly reduced in L . donovani-infected macrophages and required de novo transcription . The induction of SOCS3 provides a potent inhibitory mechanism by which intracellular microorganisms may suppress macrophage activation and interfere with the host immune response.

J Microbiol Methods, 2003 May, 53(2), 211 - 9
Analysis of environmental microbial communities by reverse sample genome probing; Greene EA et al.; Development of fast and accurate methods for monitoring environmental microbial diversity is one of the great challenges in microbiology today . Oligonucleotide probes based on 16S rRNA sequences are widely used to identify bacteria in the environment . However, the successful development of a chip of immobilized 16S rRNA probes for identification of large numbers of species in a single hybridization step has not yet been reported . In reverse sample genome probing (RSGP), labelled total community DNA is hybridized to arrays in which genomes of cultured microorganisms are spotted on a solid support in denatured form . This method has provided useful information on changes in composition of the cultured component of microbial communities in oil fields, the soil rhizhosphere, hydrocarbon-contaminated soils and acid mine drainage sites . Applications and limitations of the method, as well as the prospects of extending RSGP to cover also the as yet uncultured component of microbial communities, are evaluated.

J Microbiol Methods, 2003 May, 53(2), 175 - 83
Molecular methods for the assessment of bacterial viability; Keer JT et al.; A significant number of pathogenic microorganisms can be found in environmental reservoirs (air, water, soil) . It is important to assess the viability status of these organisms to determine whether they pose a threat to public health . Classical methods for determining viability are time consuming . Hence, molecular methods have been developed to address this problem . Molecular methods offer speed, sensitivity and specificity . Both DNA and RNA have been analysed using molecular amplification methods such as polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR) and nucleic acid sequence-based amplification (NASBA) . However, due to the variable persistence of nucleic acids in cells post-death, the correlation between presence of DNA and RNA and viability is not clear-cut . Similarly, the choice of target and sensitivity of the method can significantly affect the validity of the viability assay . This review assesses the molecular methods currently available and evaluates their ability to assess cell viability with emphasis on environmental pathogens.

J Microbiol Methods, 2003 May, 53(2), 157 - 64
PCR-based detection of non-indigenous microorganisms in 'pristine' environments; Baker GC et al.; PCR-based technologies are widely employed for the detection of specific microorganisms, and may be applied to the identification of non-indigenous microorganisms in 'pristine' environments . For 'pristine' environments such as those found on the Antarctic continent, the application of these methods to the assessment of environmental contamination from human activities must be treated with caution . Issues such as the possibility of non-human dispersal of organisms, stability and survival of non-indigenous organisms in vivo, the sensitivity, reproducibility and specificity of the PCR process (and particularly primer design) and the sampling regime employed must all be considered in detail . We conclude that despite these limitations, PCR and related technologies offer enormous scope for assessment of both natural and non-indigenous microbial distributions.

Nahrung, 2003 Feb, 47(1), 6 - 10
Mycotoxin production in wheat grains by different Aspergilli in relation to different relative humidities and storage periods; Atalla MM et al.; Four different Aspergilli (Aspergillus oryzae, A . parasiticus, A . terreus and A . versicolor) were grown on wheat grains underdifferent degrees of relative humidity 14, 50, 74, 80 and 90% . Samples of wheat grains were taken monthly for a period of six months and examined for mycotoxin production . A . oryzae was found to produce aflatoxins B1, B2, zearalenone, DON and T-2 toxins under elevated degrees of humidity and prolonged periods of storage . A . parasiticus produced aflatoxins B1, G1, NIV, DON and T-2 toxins in high concentrations during a period of not more than three months storage at 14% relative humidity; at an increased level of relative humidity of 74% ochratoxin A, zearalenone and sterigmatocystin were also produced at high levels . The isolate was drastic in toxin production . A . terrus produced toxins at 14% relative humidity (aflatoxin G2 and DON) at levels much higher than any other prevalent degrees of humidity . A . versicolor is highly sensitive to relative humidity and grain moisture content It produced aflatoxins B1, G1, NIV and DON at a relative humidity of 50% and another toxins (aflatoxin G2, ochratoxins A, B and zearalenone) at 74% . The microorganism can be considered a trichothecene producer under suitable relative humidity.

Bioresour Technol, 2003 Jan, 86(2), 157 - 64
Analysis of the logistic function model: derivation and applications specific to batch cultured microorganisms; Wachenheim DE et al.; Mathematical models are useful for describing microbial growth, both in natural ecosystems and under research conditions . To this end, a rate expression that accounted for depletion of nutrients was used to derive the logistic function model for batch cultures . Statistical analysis was used to demonstrate the suitability of this model for growth curve data . Two linear forms of the model and two procedures for calculating growth rate constants were derived to facilitate statistical evaluation of growth curves . The procedures for calculating growth rate constants were found to be useful for calculation of growth rate constants at each time point, or for estimating growth rate constants from early growth curve data . The utility of the logistic function model and its alternative forms is discussed with respect to planning experiments, analyzing growth curves for the effects of factors other than nutrient limitation, and developing more complete descriptions of cell proliferation.

Tree Physiol, 1999 Apr, 19(4_5), 313 - 320
Rhizosphere feedbacks in elevated CO(2); Cheng W; Understanding rhizosphere processes in relation to increasing atmospheric CO(2) concentrations is important for predicting the response of forest ecosystems to environmental changes, because rhizosphere processes are intimately linked with nutrient cycling and soil organic matter decomposition, both of which feedback to tree growth and soil carbon storage . Plants grown in elevated CO(2) substantially increase C input to the rhizosphere . Although it is known that elevated CO(2) enhances rhizosphere respiration more than it enhances root biomass, the fate and function of this extra carbon input to the rhizosphere in response to elevated CO(2) are not clear . Depending on specific plant and soil conditions, the increased carbon input to the rhizosphere can result in an increase, a decrease, or no effect on soil organic matter decomposition and nutrient mineralization . Three mechanisms may account for these inconsistent results: (1) the "preferential substrate utilization" hypothesis; (2) the "priming effect" hypothesis; and (3) the "competition" hypothesis, i.e., competition for mineral nutrients between plants and soil microorganisms . A microbial growth model is developed that quantitatively links the increased rhizosphere input in response to elevated CO(2) with soil organic matter decomposition . The model incorporates the three proposed mechanisms, and simulates the complexity of the rhizosphere processes . The model also illustrates mechanistically the interactions among nitrogen availability, substrate quality, and microbial dynamics when the system is exposed to elevated CO(2).

Tree Physiol, 1998 Apr, 18(4), 265 - 270
Variations in the secondary metabolite camptothecin in relation to tissue age and season in Camptotheca acuminata; Liu Z et al.; We investigated variation in concentration of the secondary metabolite, camptothecin (CPT), in relation to leaf, branch and tree age, season, and leaf drying method in Camptotheca acuminata Decaisne saplings . Younger leaves contained higher CPT concentrations than older leaves . Within a branch, there was a linear decline in CPT concentration from leaves at the apex of the branch down to Leaf 7 . Comparing leaves of similar age, those from younger trees had higher CPT concentrations than those from older trees . Over the course of the growing season, there was a steady decline of 11% per month in leaf CPT concentration . Branches showed a similar seasonal decline in CPT concentration to leaves; however, the rate of decline was threefold greater in leaves than in branches . Freeze-dried tissues had a 27% higher CPT concentration than oven- or air-dried tissues, suggesting that oven- and air-drying caused degradation of CPT . The decline in CPT concentration with tissue aging may reflect a genetically determined mechanism whereby, in young trees, chemicals serve as a first line of defense against attacks by herbivores and pathogenic microorganisms until other mechanisms are developed and deployed . We hypothesize that chemical defense mechanisms are programmed for early ontogenic stages, whereas they are induced by biotic and abiotic factors during later ontogenic stages.

J Chromatogr B Analyt Technol Biomed Life Sci, 2003 Mar 25, 786(1-2), 153 - 9
Cloning, expression and two-step purification of recombinant His-tag enhanced green fluorescent protein over-expressed in Escherichia coli; Dieryck W et al.; In this report, we describe a two-step chromatographic procedure for the purification of His-tag EGFP by immobilized metal affinity expanded bed adsorption (IMAEBA) as the capture step and size exclusion chromatography as the polishing step . The use of proteins including a histidine-tag facilitates their subsequent purification after expression in many microorganisms . This meets the needs of scientific researchers as well as industrialists in purifying recombinant proteins . The procedure described allowed the obtention of 230 mg pure EGFP from 1 l simple batch culture with a recovery of 90%.

Plant Mol Biol, 2003 Mar, 51(4), 471 - 81
Molecular characterization of a novel senescence-associated gene SPA15 induced during leaf senescence in sweet potato; Yap MN et al.; The structure and expression of a novel senescence-associated gene (SPA15) of sweet potato were characterized . The protein coding region of the gene consists of 13 exons encoding 420 amino acids . Apparent homologues of this sweet potato gene are found in a variety of dicot and monocot plants, but not in animals or microorganisms . Examination of the expression patterns of the SPA15 gene in sweet potato reveals that the transcripts of SPA15 are specifically induced in the senescing leaves, and the temporal profile of SPA15 protein accumulation is correlated with that of SPA15 transcripts . Studies on the distribution of SPA15 homologue in rice plants also indicate that SPA15 homologue is up-regulated specifically in senescing rice leaves . Treatment of detached sweet potato leaves with phytohormones including ethylene, methyl jasmonate, salicylic acid and abscisic acid resulted in a high-level induction of SPA15 . Immunoelectron microscopic analysis demonstrates that SPA15 is specifically associated with the cell wall . The potential role for SPA15 during leaf senescence is discussed.

Naturwissenschaften, 2003 Mar, 90(3), 136 - 40 Epub 2003 Feb 12.
Microbial spheres: a novel cyanobacterial-diatom symbiosis; Brehm U et al.; Cyanobacteria, algae and bacteria are widespread inhabitants of North Sea microbial mats . Our studies of these populations showed uncommon modes of living and extraordinary structures, which have not been described before . The structures are spherical objects covering a community of cyanobacteria, diatoms and bacteria . The cultivation of these communities in the laboratory and intensive observations of their exceptional movement has led to some spectacular findings . The sphere formations go through different phases with variation in the dominance of different microorganisms . The role of the bacteria is the most important in the first phase, and can be increased by the addition of signal substances . Spheres surrounded by envelopes of unknown composition and permeability appear, with numerous bacteria and sporadic diatoms inside . Then the cyanobacteria penetrate the spheres and arrange themselves at the surface . The communities proliferate over some weeks and are finally released . Laboratory expositions of the microbial communities to different parameters pinpoint the limits of sphere formation . The metabolic products of the sphere communities are concentrated in the spheres and lead to a different kind of compound compared with the surrounding environment . In this way, the microbial communities strongly influence the structure of the sediments . Uncommon circular structures, which develop into spheres between 0.08 and 3 mm in size were found in subcultures of non-axenic filamentous cyanobacteria enrichments from North Sea microbial mats . These filamentous cyanobacteria ( Phormidium sp.) together with associated benthic diatoms of the genus Navicula and associated heterotrophic bacteria were held as reproducible synergistic cultures . Phormidium sp . filaments tightly intertwined with each other, formed the surface of the spheres, trapping diatoms inside . The formation of "spheres" was the result of radial and synchronous movements of the cyanobacteria . In old cultures, the direction of the cyanobacterial movement has turned in the opposite direction, away from the sphere . The integrity of large "spheres" was influenced by chemotactic phenomena and maintained by some type of trichome-trichome interaction . This suggests the presence of metabolic secondary products, which attract cyanobacteria and influence their movement in a form of chemotactic response.

Biochim Biophys Acta, 2003 Mar 10, 1610(2), 219 - 28
Relationship between cholesterol trafficking and signaling in rafts and caveolae; Fielding CJ et al.; Caveolae and lipid rafts are two distinct populations of free cholesterol, sphingolipid (FC/SPH)-rich cell surface microdomains . They differ in stability, shape, and the presence or absence of caveolin (present in caveolae) or GPI-anchored proteins (enriched in lipid rafts) . In primary cells, caveolae and rafts support the assembly of different signaling complexes, though signal transduction from both is strongly dependent on the presence of FC . It was initially thought that FC promoted the formation of inactive reservoirs of signaling proteins . Recent data supports the concept of a more dynamic role for FC in caveolae and probably, also lipid rafts . It is more likely that the FC content of these domains is actively modulated as protein complexes are formed and, following signal transduction, disassembled . In transformed cell lines with few caveolae, little caveolin and a preponderance of rafts, complexes normally assembled on caveolae may function in rafts, albeit with altered kinetics . However, caveolae and lipid rafts appear not to be interconvertible . The presence of non-caveolar pools of caveolin in recycling endosomes (RE), the trans-Golgi network (TGN) and in mobile chaperone complexes is now recognized . A role in the uptake of microorganisms by cells ascribed to caveolae now seems more likely to be mediated by cell surface rafts.

J Dairy Sci, 2003 Feb, 86(2), 538 - 45
Gene expression of immunologically important factors in blood cells, milk cells, and mammary tissue of cows; Pfaffl MW et al.; Cytokines, eicosanoids, and lactoferrin are involved in the mammary gland's immune response to invading microorganisms . The goal of this work was to investigate the synthesis of these immunologically important factors in somatic milk cells, blood cells, and mammary tissue of cows with different somatic cell count levels, i.e., different immunological activity . On the level of mRNA expression, the cytokine tumor necrosis factor alpha (TNFalpha), lactoferrin (Lf), and specific key enzymes of leukotriene and prostaglandin biosynthesis, 5-lipoxygenase (5-LO), and cyclooxygenase-1 (COX-1) and -2 (COX-2), respectively, were determined . All 15 experimental cows were clinically healthy with no visible mammary disease . Eight cows were defined as control group with all quarters <150,000 cells/ml (C), whereas seven cows had partially elevated quarter somatic cell counts, with at least one quarter >150,000 cells/ml (H) and one quarter <150,000 cells/ml (L) . Total quarter milk from one quarter of control group and from two quarters of cows with partially elevated cell counts (one of H and one of L) was collected at one milking and a blood sample was taken simultaneously . In addition, mammary tissue samples were taken from the respective quarters on the following day during slaughter . Total RNA from milk, blood, and tissue cells was isolated and reverse transcription and quantitative polymerase chain reaction was carried out . All factors investigated were not significantly different between groups in blood cells and between C and L quarters in milk cells and mammary tissue . TNFalpha and COX-2 mRNA expression was higher in milk cells and mammary tissue of H than in L quarters, except for COX-2 in mammary tissue . Generally, TNFalpha and COX-2 showed their highest expression in milk cells, 5-LO in blood cells, whereas lactoferrin was mainly expressed by the mammary tissue . COX-1 was similarly expressed in all tested samples.

J Bacteriol, 2003 Apr, 185(7), 2096 - 103
OmcB, a c-type polyheme cytochrome, involved in Fe(III) reduction in Geobacter sulfurreducens; Leang C et al.; Microorganisms in the family Geobacteraceae are the predominant Fe(III)-reducing microorganisms in a variety of subsurface environments in which Fe(III) reduction is an important process, but little is known about the mechanisms for electron transport to Fe(III) in these organisms . The Geobacter sulfurreducens genome was found to contain a 10-kb chromosomal duplication consisting of two tandem three-gene clusters . The last genes of the two clusters, designated omcB and omcC, encode putative outer membrane polyheme c-type cytochromes which are 79% identical . The role of the omcB and omcC genes in Fe(III) reduction in G . sulfurreducens was investigated . OmcB and OmcC were both expressed during growth with acetate as the electron donor and either fumarate or Fe(III) as the electron acceptor . OmcB was ca . twofold more abundant under both conditions . Disrupting omcB or omcC by gene replacement had no impact on growth with fumarate . However, the OmcB-deficient mutant was greatly impaired in its ability to reduce Fe(III) both in cell suspensions and under growth conditions . In contrast, the ability of the OmcC-deficient mutant to reduce Fe(III) was similar to that of the wild type . When omcB was reintroduced into the OmcB-deficient mutant, the capacity for Fe(III) reduction was restored in proportion to the level of OmcB production . These results indicate that OmcB, but not OmcC, has a major role in electron transport to Fe(III) and suggest that electron transport to the outer membrane is an important feature in Fe(III) reduction in this organism.

Lett Appl Microbiol, 2003, 36(4), 222 - 6
Molecular fingerprinting of Helicobacter pylori strains from duodenal ulcer patients; Cellini L et al.; AIMS: To characterize the molecular fingerprinting of Helicobacter pylori population isolated in duodenal ulcer patients treated with triple therapy . METHODS AND RESULTS: Gastric biopsy specimens from corpus and antrum, were cultured for H . pylori isolation . Helicobacter pylori eradication was evaluated after 4 and 16 weeks . DNAs of all isolates were characterized by random amplified polymorphic DNA typing and cagA gene was also detected . After the therapy, five patients harboured the microorganism at 4 weeks and two of them remained H . pylori positive at 16 weeks . The analysis of DNA fingerprinting of strains isolated from antrum and corpus of patients susceptible to treatment, showed similar patterns . Instead, when the therapy was not effective, strains isolated from sequential biopsies from initial and after 4 and 16 weeks, showed distinct fingerprintings and retained the cagA status, over time . CONCLUSIONS: The drugs used for therapy could exercise an effect in genotypical rearrangement among H . pylori cells . SIGNIFICANCE AND IMPACT OF THE STUDY: The variableness among H . pylori strains represents a way to challenge environmental stress.

Anal Chem, 2003 Mar 1, 75(5), 1039 - 48
Use of artificial neural networks for the accurate prediction of peptide liquid chromatography elution times in proteome analyses; Petritis K et al.; The use of artificial neural networks (ANNs) is described for predicting the reversed-phase liquid chromatography retention times of peptides enzymatically digested from proteome-wide proteins . To enable the accurate comparison of the numerous LC/MS data sets, a genetic algorithm was developed to normalize the peptide retention data into a range (from 0 to 1), improving the peptide elution time reproducibility to approximately 1% . The network developed in this study was based on amino acid residue composition and consists of 20 input nodes, 2 hidden nodes, and 1 output node . A data set of approximately 7000 confidently identified peptides from the microorganism Deinococcus radiodurans was used for the training of the ANN . The ANN was then used to predict the elution times for another set of 5200 peptides tentatively identified by MS/MS from a different microorganism (Shewanella oneidensis) . The model was found to predict the elution times of peptides with up to 54 amino acid residues (the longest peptide identified after tryptic digestion of S . oneidensis) with an average accuracy of approximately 3% . This predictive capability was then used to distinguish with high confidence isobar peptides otherwise indistinguishable by accurate mass measurements as well as to uncover peptide misidentifications . Thus, integration of ANN peptide elution time prediction in the proteomic research will increase both the number of protein identifications and their confidence.

Science, 2003 Mar 28, 299(5615), 2076 - 9 Epub 2003 Mar 13.
Pyogenic bacterial infections in humans with IRAK-4 deficiency; Picard C et al.; Members of the Toll-like receptor (TLR) and interleukin-1 receptor (IL-1R) superfamily share an intracytoplasmic Toll-IL-1 receptor (TIR) domain, which mediates recruitment of the interleukin-1 receptor-associated kinase (IRAK) complex via TIR-containing adapter molecules . We describe three unrelated children with inherited IRAK-4 deficiency . Their blood and fibroblast cells did not activate nuclear factor kappaB and mitogen-activated protein kinase (MAPK) and failed to induce downstream cytokines in response to any of the known ligands of TIR-bearing receptors . The otherwise healthy children developed infections caused by pyogenic bacteria . These findings suggest that, in humans, the TIR-IRAK signaling pathway is crucial for protective immunity against specific bacteria but is redundant against most other microorganisms.

J Biol Chem, 2003 May 23, 278(21), 19463 - 72 Epub 2003 Mar 12.
Structures of shikimate dehydrogenase AroE and its Paralog YdiB . A common structural framework for different activities; Michel G et al.; Shikimate dehydrogenase catalyzes the fourth step of the shikimate pathway, the essential route for the biosynthesis of aromatic compounds in plants and microorganisms . Absent in metazoans, this pathway is an attractive target for nontoxic herbicides and drugs . Escherichia coli expresses two shikimate dehydrogenase paralogs, the NADP-specific AroE and a putative enzyme YdiB . Here we characterize YdiB as a dual specificity quinate/shikimate dehydrogenase that utilizes either NAD or NADP as a cofactor . Structures of AroE and YdiB with bound cofactors were determined at 1.5 and 2.5 A resolution, respectively . Both enzymes display a similar architecture with two alpha/beta domains separated by a wide cleft . Comparison of their dinucleotide-binding domains reveals the molecular basis for cofactor specificity . Independent molecules display conformational flexibility suggesting that a switch between open and closed conformations occurs upon substrate binding . Sequence analysis and structural comparison led us to propose the catalytic machinery and a model for 3-dehydroshikimate recognition . Furthermore, we discuss the evolutionary and metabolic implications of the presence of two shikimate dehydrogenases in E . coli and other organisms.

Nat Prod Rep, 2003 Feb, 20(1), 1 - 48
Marine natural products; Blunt JW et al.; This review covers the literature published in 2001 for marine natural products, with 497 citations (373 for the period January to December 2001) and includes 793 compounds isolated from marine microorganisms and phytoplankton, green algae, brown algae, red algae, sponges, coelenterates, bryozoans, molluscs, tunicates and echinoderms . The emphasis is on new compounds and new stereochemical assignments (683 for 2001), together with relevant biological activities, source organisms and country of origin . Syntheses that confirm or revise structures or stereochemistries have been included (95), including any first total synthesis of a marine natural product.

Indian J Exp Biol, 2002 Mar, 40(3), 319 - 24
Bioleaching of copper from chalcopyrite ore by fungi; Rao DV et al.; Microorganisms have been geologically active in mineral formation, mineral diagenesis and sedimentation via direct action of their enzymes or indirectly through chemical action of their metabolic products . This property of microorganisms is being harnessed during the recent years for extraction of metals from their ores, especially from low-grade ores . In the present study bioleaching of copper from its low-grade chalcopyrite ore using 26 isolates of acidophilic fungi is reported . Most of these fungal strains belonged to the genera Aspergillus, Penicillium and Rhizopus . The leaching experiments were conducted in Czepek Dox minimal medium containing 1% (100 mesh) ore with shaking at room temperature for 20 days . Out of these, 4 isolates exhibited significant bioleaching activities . Maximum leaching of copper (78 mg/L) was observed with Aspergillus flavus (DSF-8) and Aspergillus niger (DOF-1) . Nutritional and environmental conditions for optimum bioleaching were standardized . Present study indicates the usefulness of acidophilic fungi in bioleaching of copper from its low-grade ores.

Rheum Dis Clin North Am, 2003 Feb, 29(1), 163 - 84
Infections in systemic connective tissue diseases: systemic lupus erythematosus, scleroderma, and polymyositis/dermatomyositis; Juarez M et al.; In SLE, scleroderma, and PM/DM, infections are important causes of morbidity and mortality . This increased risk of developing infections is the result of immune abnormalities and of organ system manifestations associated with these diseases and their treatments . Common bacteria are responsible for most mild and lethal infections; however, opportunistic microorganisms cause death in some patients, particularly in those receiving high doses of corticosteroid and immunosuppressive therapy . Various viral and fungal infections also contribute to the morbidity and mortality associated with these diseases . Regardless of the cause of infections, adequate and prompt recognition and proper treatment of the infected patient are imperative . Thus, patients with these diseases, especially when receiving high doses of corticosteroids and immunosuppressive therapy, need to be monitored closely for these infections . This care and concern is necessary to ensure optimal patient outcomes, both in terms of morbidity and mortality.

Recenti Prog Med, 2003 Jan, 94(1), 12 - 5
{Infective endocarditis as predisposing factor to recurrence}; Francavilla R et al.; In this paper we communicate our experience with 60 patients who suffered two or more episodes of infectious endocarditis (IE), selected from 1053 cases of IE defined in accordance with Duke's Hospital criteria . Relapsing IE was diagnosed when episodes occurred with at least a 6 month interval between them or when, even if the time distance was shorter, they were caused by different microorganisms . Sixty-four cases of relapsing IE were observed . In each patient we considered age, sex, presence of risk-factors for IE, time laps from the onset of symptomatic disease to diagnosis, time interval between the two or more IE episodes, aetiology, endocarditic location, echocardiographic elements, therapy, complications and final outcome were considered . We conclude, agreeing with other authors, that infectious endocarditis may rightfully be considered a risk factor for further IE episodes and that, for this reason, an adequate prophylactic therapy must be given.

Parasitol Res, 2003 Mar, 89(5), 381 - 6 Epub 2002 Nov 30.
Immunohistochemical/immunogold detection and distribution of the endosymbiont Wolbachia of Dirofilaria immitis and Brugia pahangi using a polyclonal antiserum raised against WSP (Wolbachia surface protein); Kramer LH et al.; Intracellular bacteria in filarial nematodes were described as early as the 1970s, yet it was only with the work on Dirofilaria immitis, the agent of canine and feline heartworm disease, that these microorganisms were identified as belonging to Wolbachia, a genus known for encompassing bacteria infecting insects and other arthropods . The implications for the presence of intracellular bacteria in filarial nematodes is now the subject of intense research, particularly regarding their role in the immunology and pathogenesis of disease in infected humans and animals and as a possible target for therapy . Here, the authors report results on the immunohistochemical and immunogold staining of Wolbachia in D . immitis and Brugia pahangi using polyclonal antibodies raised against the recombinant Wolbachia surface protein (WSP) . The bacteria were present in the lateral hypodermal chords of both male and female worms and in the reproductive tract of adult females (oocytes, morulae, microfilariae) . In D . immitis and B . pahangi from animals treated with tetracycline, positive staining was observed in the lateral chords of adult males and females, but was absent from the oocytes and morulae . These results indicate that Wolbachia endosymbionts can be identified immunohistochemically with anti-WSP polyclonal antibodies, that their distribution matches that already described for Wolbachia of other filarial worms, and that antibiotic treatment may impede the vertical transmission of these bacteria . Unequivocal detection of Wolbachia is essential for the study of this symbiont, in particular to monitor the effects of antibiotic treatment on worms . The use of a specific marker for bacteria in their nematode hosts represents an extremely useful tool in evaluating the pathogenic role and the effect of antibiotic treatment on these potential targets in the control of filarial disease.

Zh Mikrobiol Epidemiol Immunobiol, 2003 Jan-Feb, (1), 80 - 9
{Use of the bioluminescent method for the determination of bacterial adenosinetriphosphate (ATP-metry) in microbiology}; Kuzikov AN et al.; The attention of a wide circle of specialists has recently been attracted by different methods for rapid determination of pathogenic microorganisms in biological specimens, environmental objects and foodstuffs, as well as in cases of possible acts of bioterrorism . In this respect the bioluminescent method for determination of adenosine triphosphate (ATP) contained in microbial cells is of interest . The method is based on the interaction ATP, luciferase and luciferin, accompanied by giving off energy in the form of light emission . When compared with routine methods, the use of this method considerably reduces the duration of the analysis, and its high sensitivity is comparable with that of the polymerase chain reaction . In this review the data on the prospects of the practical use of the bioluminescent method of ATP-metry are presented.

Folia Microbiol (Praha), 2002, 47(6), 672 - 6
Purine degradation in the edible mushroom Agaricus bisporus; de Windt FE et al.; Agaricus bisporus is able to use urate, allantoin, allantoate, urea and alloxanate as nitrogen sources for growth . The presence of urate oxidase, allantoinase, ureidoglycolase and urease activities, both in fruit bodies and mycelia, points to a degradative pathway for urate similar to that found in various microorganisms . So far all efforts to demonstrate the enzyme responsible for allantoate degradation failed . A urease inhibitor appeared to be present in cell-free extracts from fruit bodies.

Eur J Clin Microbiol Infect Dis, 2003 Feb, 22(2), 99 - 105 Epub 2003 Feb 15.
Viral and bacterial DNA in carotid atherosclerotic lesions; Watt S et al.; Atherosclerosis is a major health problem in industrialised countries . Several studies have suggested an association exists between certain microorganisms and the development of atherosclerosis . The aim of the study presented here was to assess the presence of viral or bacterial DNA in carotid atherosclerotic lesions . Nucleic acids were extracted from 18 carotid atherosclerotic lesions that had been collected surgically . Polymerase chain reaction was used to screen for specific genomic DNA from Chlamydia pneumoniae, cytomegalovirus and herpes simplex virus types 1 and 2 . An original approach, based on the amplification by PCR of conserved bacterial 16S rDNA nucleotide sequences was also used to detect any bacterial species . The amplification product was identified by sequencing . Chlamydia pneumoniae, cytomegalovirus and herpes simplex 2 DNA were not detected in any of the samples . Herpes simplex 1 DNA was detected in 3 of the 18 samples . Genes encoding bacterial 16S rRNA were amplified and sequenced in eight atherosclerotic lesions . DNA sequences were identified by comparison with sequences registered in the GenBank database . These eight carotid atherosclerotic lesions were shown to contain several bacterial species belonging to human flora or the environment . The exact role of these microorganisms in the genesis or development of the atherosclerotic lesions remains unclear, but they may increase the inflammatory process or be an epiphenomenon.

Annu Rev Nutr, 2003, 23, 315 - 44 Epub 2003 Mar 04.
Challenges and approaches to reducing foodborne illness; Woteki CE et al.; Microorganisms have successfully adapted to changes in food production, processing, and preservation techniques, resulting in a number of new and emerging foodborne pathogens and the re-emergence of organisms that have been problematic in the past . To protect public health, science must meet the challenges that result from the remarkable adaptability of foodborne pathogens . However, not all of the challenges of preventing foodborne illness reside in the realm of science . Food safety policy must evolve in response to new scientific understanding of hazards in the food supply and an ever-changing food processing industry . The laws, regulations, and organizations comprising the food safety system frequently lag behind current scientific knowledge of the risks posed by foodborne pathogens . Future systemic changes to enhance food safety will require better understanding of risks associated with specific pathogens occurring in the food supply and the costs and benefits of implementing mitigation strategies.

Clin Diagn Lab Immunol, 2003 Mar, 10(2), 216 - 20
Simple method to distinguish between primary and secondary C3 deficiencies; Pereira de Carvalho Florido M et al.; Due to the increasing numbers of reported clinical cases of complement deficiency in medical centers, clinicians are now more aware of the role of the complement system in the protection against infections caused by microorganisms . Therefore, clinical laboratories are now prepared to perform a number of diagnostic tests of the complement system other than the standard 50% hemolytic component assay . Deficiencies of alternative complement pathway proteins are related to severe and recurrent infections; and the application of easy, reliable, and low-cost methods for their detection and distinction are always welcome, notably in developing countries . When activation of the alternative complement pathway is evaluated in hemolytic agarose plates, some but not all human sera cross-react to form a late linear lysis . Since the formation of this linear lysis is dependent on C3 and factor B, it is possible to use late linear lysis to routinely screen for the presence of deficiencies of alternative human complement pathway proteins such as factor B . Furthermore, since linear lysis is observed between normal human serum and primary C3-deficient serum but not between normal human serum and secondary C3-deficient serum caused by the lack of factor H or factor I, this assay may also be used to discriminate between primary and secondary C3 deficiencies.

OMICS, 2002, 6(4), 305 - 30
Carbon sequestration in Synechococcus Sp.: from molecular machines to hierarchical modeling; Heffelfinger GS et al.; The U.S . Department of Energy recently announced the first five grants for the Genomes to Life (GTL) Program . The goal of this program is to "achieve the most far-reaching of all biological goals: a fundamental, comprehensive, and systematic understanding of life." While more information about the program can be found at the GTL website , this paper provides an overview of one of the five GTL projects funded, "Carbon Sequestration in Synechococcus Sp.: From Molecular Machines to Hierarchical Modeling." This project is a combined experimental and computational effort emphasizing developing, prototyping, and applying new computational tools and methods to elucidate the biochemical mechanisms of the carbon sequestration of Synechococcus Sp., an abundant marine cyanobacteria known to play an important role in the global carbon cycle . Understanding, predicting, and perhaps manipulating carbon fixation in the oceans has long been a major focus of biological oceanography and has more recently been of interest to a broader audience of scientists and policy makers . It is clear that the oceanic sinks and sources of CO(2) are important terms in the global environmental response to anthropogenic atmospheric inputs of CO(2) and that oceanic microorganisms play a key role in this response . However, the relationship between this global phenomenon and the biochemical mechanisms of carbon fixation in these microorganisms is poorly understood . The project includes five subprojects: an experimental investigation, three computational biology efforts, and a fifth which deals with addressing computational infrastructure challenges of relevance to this project and the Genomes to Life program as a whole . Our experimental effort is designed to provide biology and data to drive the computational efforts and includes significant investment in developing new experimental methods for uncovering protein partners, characterizing protein complexes, identifying new binding domains . We will also develop and apply new data measurement and statistical methods for analyzing microarray experiments . Our computational efforts include coupling molecular simulation methods with knowledge discovery from diverse biological data sets for high-throughput discovery and characterization of protein-protein complexes and developing a set of novel capabilities for inference of regulatory pathways in microbial genomes across multiple sources of information through the integration of computational and experimental technologies . These capabilities will be applied to Synechococcus regulatory pathways to characterize their interaction map and identify component proteins in these pathways . We will also investigate methods for combining experimental and computational results with visualization and natural language tools to accelerate discovery of regulatory pathways . Furthermore, given that the ultimate goal of this effort is to develop a systems-level of understanding of how the Synechococcus genome affects carbon fixation at the global scale, we will develop and apply a set of tools for capturing the carbon fixation behavior of complex of Synechococcus at different levels of resolution . Finally, because the explosion of data being produced by high-throughput experiments requires data analysis and models which are more computationally complex, more heterogeneous, and require coupling to ever increasing amounts of experimentally obtained data in varying formats, we have also established a companion computational infrastructure to support this effort as well as the Genomes to Life program as a whole.

Prikl Biokhim Mikrobiol, 2003 Jan-Feb, 39(1), 25 - 30
{Estimation of the hydrocarbon-oxidizing activity of microorganisms}; Petrikevich SB et al.; A method allowing the microorganisms growing on substrates with a low water solubility (oil, fuel oil, resins, and asphalthenes) to be isolated and counted was developed . An advantage of the method is its ability to estimate the oil-utilizing activity of each strain visually according to the decolorized zones formed during its growth on oil products . The sizes of these zones indicate which oil-degrading strain is most active.

Prikl Biokhim Mikrobiol, 2003 Jan-Feb, 39(1), 5 - 24
{Deceleration of vital activity as a universal biochemical mechanism ensuring adaptation of microorganisms to stress factors: a review}; Feofilova EP; The review deals with a major problem of contemporary biology, i.e., the strategy of persistence of organisms affected by stressors . The primary mechanism of biochemical adaptation of cells is their transition to a specific surviving state, which is characterized by deceleration of vital activity (DVA), a process related to decreased metabolism . According to the latest classification, there are four DVA types: quiescence (endogenous or exogenous), cryptobiosis, anabiosys, and latent life . These states are viewed from a new standpoint, as depending on both the stage of growth and development of the organism and the type of stressor . A brief characterization of cryptobiosis and anabiosys is provided . Quiescence, which is a specific type of DVA, is characterized in greater detail, with emphasis on differences between fungi and bacteria . Endogenous and exogenous quiescence is compared in two types of cells (spores) . Changes in the composition of quiescent cells (spores) as compared to their vegetating counterparts involve lipids (membrane-associated and neutral), cytosolic carbohydrates (which serve as a reserve supply of nutritives or act as protectors), water content, RNA/protein synthesis, and cyclic nucleotides (e.g., cAMP) . Biochemical mechanisms controlling reversion of DVA (the transition from spores to germinating cells) are also analyzed . In conclusion, consideration is given to the role of DVA in evolution and to the implications of DVA studies for biotechnology, cryogenic preservation, utilization of interstellar space and permafrost territories.

Ying Yong Sheng Tai Xue Bao, 2002 Nov, 13(11), 1516 - 8
{Preliminary study on the growth stimulation of marine microalgae stimulated by low level of toxicant}; Huang J et al.; Cell growth of Nitzschia closterium and Platymonas subcordiformis was stimulated under 4 and 8 micrograms.L-1 anthrancene stress, respectively . The cell growth of two species was also stimulated under 30 mg.L-1 streptomycin stress . The hormesis of marine microalgae cells under low level of anthrancene and streptomycin resulted in low level of free radical and decrease of microorganisms . Furthermore, the hormesis of marine microalgae was also discussed in this paper.

Amino Acids, 2003, 24(1-2), 135 - 9
Studies on the utilization of methionine sulfoxide and methionine sulfone by rumen microorganisms in vitro; Or-Rashid MM et al.; An in vitro experiment was conducted to test the ability of mixed rumen bacteria (B), protozoa (P), and their mixture (BP) to utilize the oxidized forms of methionine (Met) e.g., methionine sulfoxide (MSO), methionine sulfone (MSO(2)) . Rumen contents were collected from fistulated goats to prepare the microbial suspensions and were anaerobically incubated at 39 degrees C for 12 h with or without MSO (1 mM) or MSO(2) (1 mM) as a substrate . Met and other related compounds produced in both the supernatants and hydrolyzates of the incubation were analyzed by HPLC . During 6- and 12-h incubation periods, MSO disappeared by 28.3 and 42.0%, 0.0 and 0.0%, and 40.6 and 62.4% in B, P, and BP suspensions, respectively . Rumen bacteria and the mixture of rumen bacteria and protozoa were capable to reduce MSO to Met, and the production of Met from MSO in BP (156.6 and 196.1 micromol/g MN) was about 17.3 and 14.1% higher than that in B alone (133.5 and 171.9 micromol/g MN) during 6- and 12-h incubations, respectively . On the other hand, mixed rumen protozoa were unable to utilize MSO . Other metabolites produced from MSO were found to be MSO(2) and 2-aminobutyric acid (2AB) in B and BP . MSO(2) as a substrate remained without diminution in all-microbial suspensions . It was concluded that B, P, and BP cannot utilize MSO(2); but MSO can be utilized by B and BP for producing Met.

Med Clin (Barc), 2003 Mar 1, 120(7), 253 - 4
{Infection due to Mycobacterium kansasii in Elche, Spain}; Rodriguez JC et al.; Background and objective: Our goal was to determine the incidence of infection due to Mycobacterium kansasii in Elche, Spain over the last 10 years and to study the activity of new antibiotics against the strains isolated.Material and methods: A retrospective study of patients and study of antibiotic susceptibility in vitro.Results: The annual incidence of infection by this microorganism was 2.66 cases/ 1,000,000 inhabitants per year . In patients with AIDS, the incidence increased to 571.4 cases/1,000,000 inhabitants per year . Gatifloxacin and moxifloxacin exhibit better in vitro activity than levofloxacin; in some strains, a synergism exists between rifampi cin/fluoroquinolones and rifampicin/linezolid.Conclusion: In Elche, infections due to M . kansasii are uncommon . It is necesary to continue evaluating the activity of fluoroquinolones and linezolid.

Mol Microbiol, 2003 Mar, 47(6), 1749 - 58
The Leopoldina international symposium on parasitism, commensalism and symbiosis--common themes, different outcome; Gross R et al.; The development of new methods, including genomics, which can even be applied to unculturable microorganisms, has significantly increased our knowledge about bacterial pathogenesis and symbiosis and, in consequence, is profoundly modifying our views on the evolution and the genetic and physiological basis of bacteria-host interactions . The presentations at this symposium revealed conceptual links between bacterial pathogenesis and symbiosis . The close co-operation of experts in both fields will result in significant synergy and new insights into basic mechanisms of bacteria-host interactions and their evolution . The meeting provided fascinating news about the genetic and metabolic consequences that the change in their lifestyle had for bacteria that developed from free-living to permanent host-associated organisms exemplified by intracellular pathogens or symbionts . In addition, surprising similarities but also striking differences between the strategies involved in the establishment of a symbiotic versus a parasitic lifestyle can be noted . In the long run, the characterization of such differences might lead to lifestyle prediction or to an evaluation of the pathogenic potential of newly isolated bacteria via the definition of genetic and/or metabolic signatures characteristic for pathogenic or symbiotic organisms . Moreover, it is expected that these investigations will lead to new strategies for the treatment or prevention of bacterial infections, or the avoidance of pathogen transmission.

Anal Chem, 2003 Feb 15, 75(4), 825 - 34
Electrophoretic behavior and potency assessment of boar sperm using a capillary electrophoresis-laser induced fluorescence system; He L et al.; The assessment of mammalian fertility, and the agents that affect it, is of increasing concern in medicinal, environmental, and agricultural science . The viability, integrity, and overall state of the male gamete (sperm) is an essential factor that must be considered in such studies . Traditional potency evaluations tend to be labor intensive and often are not precise . A CE-LIF technique for determining the viability of boar sperm was developed using the fluorescent stains SYBER-14 and propidium iodide . The buffer type, pH, ionic strength, applied voltage, and polymer additive must be optimized in order to obtain sharp peaks and accurate results . Extender solutions that are widely used in artificial insemination programs were found to be compatible and even beneficial for these CE-LIF experiments . A single viability assay takes less than 10 min, which is significantly faster than most other procedures . The compaction or focusing of the sample zone seems to be similar to that reported previously for microorganisms in CE.

Int J Hyg Environ Health, 2003 Jan, 206(1), 61 - 4
Ranking and frequency of somatic symptoms in residents near composting sites with odor annoyance; Herr CE et al.; The presented study reports the prevalence of somatic symptoms in three study samples living in the vicinity of composting plants . Microorganisms were measured in the air of the residential areas closest to the plants at the same time an epidemiological investigation was performed in the neighborhood near (150 to 1,500 m) to three plants and in corresponding control residential areas of the same district . Nine hundred and seventy-nine residents were questioned about the odor annoyance in their vicinity . An instrument measuring somatic complaints (SOMS 2 acc . Rief et al., 2001) was used to determine the unexplained somatic symptoms of the past two years and a gender-independent Total Complaint Index (TCI) was calculated . The percentages of study population reporting somatic symptoms were higher in all six samples in comparison with the German population (Rief et al., 2001) and in samples living near composting sites compared to the corresponding control samples . The study sample living close to site A (exposed to bioaerosols and odor annoyance) had the highest rates of complaints . A difference could be seen in comparison to the corresponding control sample (TCI: p{Anear vs . Acontrol} = 0.001; Mann-Whitney) . In this group breathlessness was reported more than twice as often as in the other three samples . Out of the five most frequently reported symptoms four corresponded to the five complaints the German population reports most frequently in all groups . Nausea was the fifth most reported symptom in both samples reporting annoying residential odors (Anear and Bnear) . The type of somatic symptoms reported most often was influenced little by environmental odors and medically relevant bioaerosol concentrations, except for nausea in context with annoying residential odors . As expected frequency of reporting general somatic symptoms was influenced by the perceived environment near the three composting sites . Concerning the sum of bodily complaints (TCI) though, this was only significant in the group exposed to medically relevant concentrations of residential outdoor bioaerosols, which was accompanied by high rates for breathlessness . The SOMS2 was able to mirror measurable, medically relevant environmental exposures in study groups and showed fewer changes concerning annoying or presumed environmental exposures.

Appl Environ Microbiol, 2003 Mar, 69(3), 1662 - 9
The earthworm gut: an ideal habitat for ingested N2O-producing microorganisms; Horn MA et al.; The in vivo production of nitrous oxide (N(2)O) by earthworms is due to their gut microbiota, and it is hypothesized that the microenvironment of the gut activates ingested N(2)O-producing soil bacteria . In situ measurement of N(2)O and O(2) with microsensors demonstrated that the earthworm gut is anoxic and the site of N(2)O production . The gut had a pH of 6.9 and an average water content of approximately 50% . The water content within the gut decreased from the anterior end to the posterior end . In contrast, the concentration of N(2)O increased from the anterior end to the mid-gut region and then decreased along the posterior part of the gut . Compared to the soil in which worms lived and fed, the gut of the earthworm was highly enriched in total carbon, organic carbon, and total nitrogen and had a C/N ratio of 7 (compared to a C/N ratio of 12 in soil) . The aqueous phase of gut contents contained up to 80 mM glucose and numerous compounds that were indicative of anaerobic metabolism, including up to 9 mM formate, 8 mM acetate, 3 mM lactate, and 2 mM succinate . Compared to the soil contents, nitrite and ammonium were enriched in the gut up to 10- and 100-fold, respectively . The production of N(2)O by soil was induced when the gut environment was simulated in anoxic microcosms for 24 h (the approximate time for passage of soil through the earthworm) . Anoxia, high osmolarity, nitrite, and nitrate were the dominant factors that stimulated the production of N(2)O . Supplemental organic carbon had a very minimal stimulatory effect on the production of N(2)O, and addition of buffer or ammonium had essentially no effect on the initial N(2)O production rates . However, a combination of supplements yielded rates greater than that obtained mathematically for single supplements, suggesting that the maximum rates observed were due to synergistic effects of supplements . Collectively, these results indicate that the special microenvironment of the earthworm gut is ideally suited for N(2)O-producing bacteria and support the hypothesis that the in situ conditions of the earthworm gut activate ingested N(2)O-producing soil bacteria during gut passage.

Appl Environ Microbiol, 2003 Mar, 69(3), 1408 - 16
Construction and screening of metagenomic libraries derived from enrichment cultures: generation of a gene bank for genes conferring alcohol oxidoreductase activity on Escherichia coli; Knietsch A et al.; Enrichment of microorganisms with special traits and the construction of metagenomic libraries by direct cloning of environmental DNA have great potential for identifying genes and gene products for biotechnological purposes . We have combined these techniques to isolate novel genes conferring oxidation of short-chain (C(2) to C(4)) polyols or reduction of the corresponding carbonyls . In order to favor the growth of microorganisms containing the targeted genes, samples collected from four different environments were incubated in the presence of glycerol and 1,2-propanediol . Subsequently, the DNA was extracted from the four samples and used to construct complex plasmid libraries . Approximately 100,000 Escherichia coli strains of each library per test substrate were screened for the production of carbonyls from polyols on indicator agar . Twenty-four positive E . coli clones were obtained during the initial screen . Sixteen of them contained a plasmid (pAK101 to pAK116) which conferred a stable carbonyl-forming phenotype . Eight of the positive clones exhibited NAD(H)-dependent alcohol oxidoreductase activity with polyols or carbonyls as the substrates in crude extracts . Sequencing revealed that the inserts of pAK101 to pAK116 encoded 36 complete and 17 incomplete presumptive protein-encoding genes . Fifty of these genes showed similarity to sequenced genes from a broad collection of different microorganisms . The genes responsible for the carbonyl formation of E . coli were identified for nine of the plasmids (pAK101, pAK102, pAK105, pAK107 to pAK110, pAK115, and pAK116) . Analyses of the amino acid sequences deduced from these genes revealed that three (orf12, orf14, and orf22) encoded novel alcohol dehydrogenases of different types, four (orf5, sucB, fdhD, and yabF) encoded novel putative oxidoreductases belonging to groups distinct from alcohol dehydrogenases, one (glpK) encoded a putative glycerol kinase, and one (orf1) encoded a protein which showed no similarity to any other known gene product.

Poult Sci, 2003 Feb, 82(2), 223 - 31
Effect of minerals on activity of microbial uricase to reduce ammonia volatilization in poultry manure; Kim WK et al.; Inhibition of microbial uricase in poultry manure is critical to reduce NH3 volatilization, because hydrolysis of uric acid by microbial uricase is the first step in the production of NH3 gas in poultry manure . Three experiments were conducted to evaluate the effects of minerals on uricase activity and NH3 volatilization from poultry manure . In Experiment 1, an in vitro enzyme assay was used to evaluate the effects of Zn, Cu, Mg, and Mn on the activity of microbial uricase . There were three treatments: uricase, uricase + minerals, and uricase preincubated with minerals . Uric acid concentration was measured at 293 nm with a spectrophotometer . The results indicated that Zn and Cu greatly blocked the activity of microbial uricase (>90% inhibition), whereas Mg and Mn were less inhibitory . Experiment 2 was designed to evaluate the effect of ZnSO4 on the growth of uric acid-utilizing microorganisms by an in vitro assay . There were three treatments: control, ZnSO4 (10 mM), and ZnSO4 (50 mM) . The results indicated that ZnSO4 significantly reduced the number of uric acid-utilizing microorganisms compared to the control . In Experiment 3, an NH3-trapping system was used to evaluate the effect of different levels of ZnSO4 on NH3 volatilization and nitrogen retention in poultry manure . Poultry manure (300 g) was mixed with 0, 0.15, 0.3, 1.5, 3, or 6 g ZnSO4 to create manure concentrations of Zn at 0, 0.05, 0.1, 0.5, 1, or 2% (wt/wt), respectively . The 1 and 2% ZnSO4 treatments significantly increased manure uric acid and total nitrogen retention by reducing NH3 volatilization compared to the control during the 3-wk incubation.

Chemphyschem, 2003 Feb 17, 4(2), 162 - 8
Electrochemical approaches for chemical and biological analysis on Mars; Kounaves SP; Obtaining in situ chemical data from planetary bodies such as Mars or Europa can present significant challenges . The one analytical technique that has many of the requisite characteristics to meet such a challenge is electroanalysis . Described here are three electroanalytical devices designed for in situ geochemical and biological analysis on Mars . The Mars Environmental Compatibility Assessment (MECA) was built and flight qualified for the now cancelled NASA Mars 2001 Lander . Part of MECA consisted of four "cells" containing arrays of electrochemical based sensors for measuring the ionic species in soil samples . A next-generation MECA, the Robotic Chemical Analysis Laboratory (RCAL), uses a carousel-type system to allow for greater customization of analytical procedures . A second instrument, proposed as part of the 2007 CryoScout mission, consists of a flow-through inorganic chemical analyzer (MICA) . CryoScout is a torpedo-like device designed for subsurface investigation of the stratigraphic climate record embedded in Mars' north polar cap . As the CryoScout melts its way through the ice cap, MICA will collect and analyze the meltwater for a variety of inorganics and chemical parameters . By analyzing the chemistry locked in the layers of dust, salt, and ice, geologists will be able to determine the recent history of climate, water, and atmosphere on Mars and link it to the past . Finally, electroanalysis shows its abilities in the detection of possible microorganism on Mars or elsewhere in the solar system . To identify an unknown microorganism, one that may not even use Earth-type biochemistry, requires a detection scheme which makes minimal assumptions and looks for the most general features . Recent work has demonstrated that the use of an array of electrochemical sensors which monitors the changes in a solution via electrical conductivity, pH, and ion selective electrodes, can be used to detect minute chemical perturbations caused by the growth of bacteria and with the correct methodology provide unamibiguous detection of such life forms.

Nat Mater, 2002 Nov, 1(3), 169 - 72
Biomimetic synthesis and patterning of silver nanoparticles; Naik RR et al.; The creation of nanoscale materials for advanced structures has led to a growing interest in the area of biomineralization . Numerous microorganisms are capable of synthesizing inorganic-based structures . For example, diatoms use amorphous silica as a structural material, bacteria synthesize magnetite (Fe3O4) particles and form silver nanoparticles, and yeast cells synthesize cadmium sulphide nanoparticles . The process of biomineralization and assembly of nanostructured inorganic components into hierarchical structures has led to the development of a variety of approaches that mimic the recognition and nucleation capabilities found in biomolecules for inorganic material synthesis . In this report, we describe the in vitro biosynthesis of silver nanoparticles using silver-binding peptides identified from a combinatorial phage display peptide library.

Lipids, 2002 Dec, 37(12), 1177 - 86
Evidence for multiple sterol methyl transferase pathways in Pneumocystis carinii; Zhou W et al.; The sterol composition of Pneumocystis carinii, an opportunistic pathogen responsible for life-threatening pneumonia in immunocompromised patients, was determined . Our purpose was to identify pathway-specific enzymes to impair using sterol biosynthesis inhibitors . Prior to this study, cholesterol 15 (ca . 80% of total sterols), lanosterol 1, and several phytosterols common to plants (sitosterol 31, 24alpha-ethyl and campesterol, 24alpha-methyl 30) were demonstrated in the fungus . In this investigation, we isolated all the previous sterols and many new compounds from P . carinii by culturing the microorganism in steroid-immunosuppressed rats . Thirty-one sterols were identified from the fungus (total sterol = 100 fg/cell), and seven sterols were identified from rat chow . Unusual sterols in the fungus not present in the diet included, 24(28)-methylenelanosterol 2; 24(28)E-ethylidene lanosterol 3; 24(28)Z-ethylidene lanosterol 4; 24beta-ethyllanosta-25(27)-dienol 5; 24beta-ethylcholest-7-enol 6; 24beta-ethylcholesterol 7; 24beta,-ethylcholesta-5,25(27)-dienol 8; 24-methyllanosta-7-enol 9; 24-methyldesmosterol 10; 24(28)-methylenecholest-7-enol 11; 24beta-methylcholest-7-enol 12; and 24beta-methylcholesterol 13 . The structural relationships of the 24-alkyl groups in the sterol side chain were demonstrated chromatographically relative to authentic specimens, by MS and high-resolution 1H NMR . The hypothetical order of these compounds poses multiple phytosterol pathways that diverge from a common intermediate to generate 24beta-methyl sterols: route 1, 1 --> 2 --> 11 --> 12 --> 13; route 2, 1 --> 2 --> 9 --> 10 --> 13; or 24beta-ethyl sterols: route 3, 1 --> 2 --> 4 --> 6 --> 7; route 4, 1 --> 2 --> 5 --> 8 --> 7 . Formation of 3 is considered to form an interrupted sterol pathway . Taken together, operation of distinct sterol methyl transferase (SMT) pathways that generate 24beta-alkyl sterols in P . carinii with no counterpart in human biochemistry suggests a close taxonomic affinity with fungi and provides a basis for mechanism-based inactivation of SMT enzyme to treat Pneumocystis pneumonia.

Ann Surg, 2003 Mar, 237(3), 408 - 16
Regional and systemic cytokine responses to acute inflammation of the vermiform appendix; Rivera-Chavez FA et al.; OBJECTIVE: To measure local (peritoneal fluid) and systemic (plasma) cytokine profiles in patients with infection-inflammation of the vermiform appendix, a relatively mild, localized inflammatory process . SUMMARY BACKGROUND DATA: The systemic host response to invading microorganisms, often termed the systemic inflammatory response syndrome (SIRS), includes changes in heart rate, respiratory rate, body temperature, and circulating white blood cell numbers . Although these changes can be induced experimentally by administering proinflammatory cytokines, the mediators that appear in the bloodstream during early, localized infection in humans have not been defined . METHODS: The authors studied 56 patients with pathologically proven appendicitis . Blood was obtained before the induction of anesthesia, when 82% of the patients met the criteria for SIRS . Peritoneal fluid (PF) was obtained by intraoperative lavage . Cytokines were measured by immunoassay . To assess the net impact of the mediators within plasma, the authors studied the ability of patient plasma to augment or suppress bacterial lipopolysaccharide (LPS) stimulation of monocytes in vitro . RESULTS: Of the proinflammatory cytokines, tumor necrosis factor-alpha was present in PF but not in plasma, interleukin (IL)-1beta and interferon-gamma were found in low concentrations in both PF and plasma, and IL-12 (p70) was detectable in plasma but not PF . In contrast, IL-6 and IL-1 receptor antagonist (IL-1ra) were the most abundant cytokines in the PF and plasma, and the concentrations of IL-4 and IL-10 were also elevated in both compartments . Patients with more severe appendicitis had higher plasma levels of IL-6 and IL-10 and lower plasma levels of IL-12 and interferon-gamma than did those with uncomplicated disease . Patient plasma inhibited LPS-induced stimulation of a monocyte cell line, and this inhibition was accentuated by complicated disease . CONCLUSIONS: As judged from the pattern of soluble cytokines in plasma and the effect of the plasma on monocyte activation by LPS, mild, localized infection can induce a systemic response that is predominantly anti-inflammatory.

J Biotechnol, 2003 Mar 20, 101(3), 241 - 52
Decolorization of azo dye using PVA-immobilized microorganisms; Chen KC et al.; A microbial consortium having a high capacity for rapid decolorization of azo dye (RED RBN) was immobilized by a phosphorylated polyvinyl alcohol (PVA) gel . The immobilized-cell beads exhibited a color removal capability of 75%, even at a high concentration of RED RBN (500 mg l(-1)) within 12 h using flask culture . The continuous operation was conducted at a hydraulic retention time (HRT) of 5-20 h in which the dye loading rate ranged from 240 to 60 mg dye h(-1) . A removal efficiency exceeding 90% was obtained at the HRT higher than 10 h . No recognizable destruction of bead appearance was observed in the 6-month operation . Examination of the mechanism of the decolorization process by cell beads indicated that it proceeded primarily by biological decolorization associated with partial adsorption of the dye onto the entrapped cells and gel matrix . Microscopic observation revealed that the microbial consortium contained in the gel beads was at least made up of three kinds of bacterial species . From the economical viewpoint, alternative cheaper nitrogen sources such as fish meal, soybean meal, pharmamedia and vita yeast powder were examined.

Biochemistry, 2003 Mar 11, 42(9), 2552 - 9
Phosphorylation triggers domain separation in the DNA binding response regulator NarL; Zhang JH et al.; DNA binding proteins of two-component signal transduction systems in microorganisms are activated by phosphorylation through an unknown mechanism . NarL is an example from the nitrate/nitrite signal transduction system of Escherichia coli . NarL consists of N- and C-terminal domains, the latter of which contains the DNA binding elements . To explore the mechanism of activation, single nitroxide side chains were introduced, one at a time, at nine different sites throughout the C-terminal domain to monitor the tertiary structure and the status of the surface in contact with the N-terminal domain . In addition, three pairs of doubly labeled proteins were prepared to monitor the interdomain distance using the magnetic dipolar interaction . The results of these site-directed spin-labeling studies reveal that phosphorylation at a distant site in the N-terminal domain triggers domain separation, likely by a hinge-bending motion . This in turn presents key elements of the C-terminal domain for docking to the DNA target in the configuration described in the recent crystal structure . The data also imply that a single conformation of unphosphorylated NarL exists in solution, and there is no detectable equilibrium between the closed and open conformations.

Pediatr Infect Dis J, 2003 Feb, 22(2), 199 - 200
Capofungin therapy for Aspergillus lung infection in a boy with chronic granulomatous disease; Sallmann S et al.; Neutrophils of patients with chronic granulomatous disease (CGD) are profoundly defective in killing microorganisms . As in patients with chemotherapy-induced neutropenia, CGD patients are highly susceptible to life-threatening invasive Aspergillus infections, and conventional antimycotic treatment is not always successful . The new drug, caspofungin, was used to treat Aspergillus lung infection in one child with CGD.

Curr Opin Drug Discov Devel, 2003 Jan, 6(1), 57 - 65
Metabolite identification in drug discovery; Watt AP et al.; Recent developments in the technologies and approaches to identify metabolites in a drug discovery environment are reviewed . Samples may be generated using either in vitro systems--typically, but not exclusively, liver subcellular fractions, such as microsomes, or whole cells, such as hepatocytes . Alternatively, metabolites are generated in vivo using excreta obtained following dosing in preclinical species . Recombinant drug metabolizing enzymes or microorganisms may offer alternate vectors . New techniques, such as the use of solid-phase microextraction, have found application in the isolation of metabolites from biological matrices . However, this is still dominated by the use of preparative chromatography, which has advanced through the use of mass-directed detection . Detection and structural elucidation by mass spectrometry have improved markedly with increases in sensitivity, allowing lower abundance metabolites to be detected, and increases in selectivity, with the use of high-resolution time-of-flight and quadrupole-time-of-flight instruments . Finally, higher field strength magnets coupled with novel probe designs and increased use of liquid chromatographic hyphenation techniques continue to drive the capabilities of nuclear magnetic resonance spectroscopy as the definitive structural elucidation tool.

Plant J, 2003 Feb, 33(4), 775 - 92
GFP-tagging of cell components reveals the dynamics of subcellular re-organization in response to infection of Arabidopsis by oomycete pathogens; Takemoto D et al.; Cytoplasmic aggregation, the rapid translocation of cytoplasm and subcellular components to the site of pathogen penetration, is one of the earliest reactions of plant cells against attack by microorganisms . We have investigated cytoplasmic aggregation during Arabidopsis-oomycete interactions . Infection by non-pathogenic Phytophthora sojae was prevented in the plant epidermal cell layer, whereas Peronospora parasitica isolates Cala2 (avirulent) and Noks1 (virulent) could both penetrate into the mesophyll cell layer . Epidermal cell responses to penetration by these oomycetes were examined cytologically with a range of transgenic Arabidopsis plants expressing Green Fluorescent Protein (GFP)-tagged cell components . These included plants containing GFP-TUA6 for visualizing microtubules, GFP-hTalin for actin microfilaments, GFP-tm-KKXX for endoplasmic reticulum (ER), and STtmd-GFP for the Golgi apparatus . In all interactions, actin microfilaments were actively re-arranged and formed large bundles in cytoplasmic strands focused on the penetration site . Aggregation of ER membrane and accumulation of Golgi bodies at the infection site were observed, suggesting that production and secretion of plant materials were activated around the penetration site . Microtubules did not become focused on the penetration site . No difference was evident between the responses of epidermal cells in the non-host, incompatible and compatible interactions . This result indicates that the induction of cytoplasmic aggregation in Arabidopsis epidermal cells was neither suppressed by the virulent strain of Peronospora, nor effective in stopping infection.

Indoor Air, 2003 Mar, 13(1), 38 - 48
'EUROPART' . Airborne particles in the indoor environment . A European interdisciplinary review of scientific evidence on associations between exposure to particles in buildings and health effects; Schneider T et al.; The relevance of particle mass, surface area or number concentration as risk indicators for health effects in non-industrial buildings has been assessed by a European interdisciplinary group of researchers (called EUROPART) by reviewing papers identified in Medline, Toxline, and OSH . Studies dealing with dermal effects or cancer or specifically addressing environmental tobacco smoke, house dust-mite, cockroach or animal allergens, microorganisms and pesticides were excluded . A total of 70 papers were reviewed, and eight were identified for the final review: Five experimental studies involving mainly healthy subjects, two cross-sectional office studies and one longitudinal study among elderly on cardiovascular effects . From most studies, no definite conclusions could be drawn . Overall, the group concluded that there is inadequate scientific evidence that airborne, indoor particulate mass or number concentrations can be used as generally applicable risk indicators of health effects in non-industrial buildings and consequently that there is inadequate scientific evidence for establishing limit values or guidelines for particulate mass or number concentrations.

Kansenshogaku Zasshi, 2002 Dec, 76(12), 1010 - 5
{Effect of the pretreatment with organic acid buffer solutions on the recovery of Legionella species from environmental water samples}; Kasuga O et al.; Anti-microbial effect of the pretreatment with various organic acid buffer solutions against co-existing microorganisms which were isolated from cooling-tower water samples along with Legionella spp . was examined . Among several buffer solutions, a 0.1 M potassium citrate-citric acid buffer solution (hereafter, citrate buffer solution, pH 2.2) hardly affected the recovery of Legionella spp., but effectively inhibited the growth of co-existing microorganisms . To evaluate the buffer action of these buffer solutions, pHs of 9 cooling-tower water samples were evaluated after addition of an equal volume of each buffer solution . When a citrate buffer solution . pH 2.2 was combined to a 200-fold concentrated solution of each cooling-tower water sample, the pH of the combined solution became 2.5 to 2.7 and maintained a stably low pH value than that (pH 3.0 to 7.4) obtained after mixture of a 0.2 M HCl-KCl buffer solution (hereafter, HCl buffer solution, pH 2.2), suggesting strong buffer action of the citrate buffer solution, pH 2.2 in the combined solutions . Furthermore, when cooling-tower water samples were pretreated with a citrate buffer solution, pH 2.2, the recovery of Legionella spp . was successful in 7 out of 9 cooling-tower water samples, suggesting 3 times higher recovery rate than that obtained by addition of a HCl buffer solution, pH 2.2 (3 out of 9 cooling-tower water samples).

Environ Int, 2003 Mar, 28(8), 771 - 8
Novel approach to monitoring of the soil biological quality; Hofman J et al.; In this study, a new approach to interpretation of results of the simple microbial biomass and respiration measurements in the soil microbiology is proposed . The principle is based on eight basal and derived microbial parameters, which are standardized and then plotted into sunray plots . The output is visual presentation of one plot for each soil, which makes possible the relative comparison and evaluation of soils in the monitored set . Problems of soil microbiology, such as the lack of benchmarking and reference values, can be avoided by using the proposed method . We found that eight parameters provide enough information for evaluation of the status of the soil microorganisms and, thus, for evaluation of the soil biological quality . The usage of rare parameters (potential respiration PR, ratio of potential and basal respiration PR/BR, biomass-specific potential respiration PR/C(bio), available organic carbon C(ext), and biomass-specific available organic carbon C(ext)/C(bio)) can be recommended, besides classical and well-known parameters (microbial biomass C(bio), basal respiration BR, metabolic coefficient qCO(2)) . The combination of basal parameters and derived coefficients can also extend our knowledge about the condition of the soil microorganisms . In monitoring the case studies presented, we observed that soils evaluated to possess good biological quality displayed generally higher values of organic carbon, total nitrogen, clay, and cation exchange capacity . The soils of good biological quality can display higher levels of contaminants . This is probably related with the higher content of organic carbon and clay in these soils.

Biosens Bioelectron, 2003 Apr, 18(4), 405 - 13
RNA biosensor for the rapid detection of viable Escherichia coli in drinking water; Baeumner AJ et al.; A highly sensitive and specific RNA biosensor was developed for the rapid detection of viable Escherichia coli as an indicator organism in water . The biosensor is coupled with protocols developed earlier for the extraction and amplification of mRNA molecules from E . coli {Anal . Biochem . 303 (2002) 186} . However, in contrast to earlier detection methods, the biosensor allows the rapid detection and quantification of E . coli mRNA in only 15-20 min . In addition, the biosensor is portable, inexpensive and very easy to use, which makes it an ideal detection system for field applications . Viable E . coli are identified and quantified via a 200 nt-long target sequence from mRNA (clpB) coding for a heat shock protein . For sample preparation, a heat shock is applied to the cells prior to disruption . Then, mRNA is extracted, purified and finally amplified using the isothermal amplification technique Nucleic acid sequence-based amplification (NASBA) . The amplified RNA is then quantified with the biosensor . The biosensor is a membrane-based DNA/RNA hybridization system using liposome amplification . The various biosensor components such as DNA probe sequences and concentration, buffers, incubation times have been optimized, and using a synthetic target sequence, a detection limit of 5 fmol per sample was determined . An excellent correlation to a much more elaborate and expensive laboratory based detection system was demonstrated, which can detect as few as 40 E . coli cfu/ml . Finally, the assay was tested regarding its specificity; no false positive signals were obtained from other microorganisms or from nonviable E . coli cells .

Chem Biol Interact, 2003 Feb 1, 143-144, 229 - 38
Properties and functional significance of Saccharomyces cerevisiae ADHVI; Larroy C et al.; The completion of the Saccharomyces cerevisiae genome project has provided the opportunity to explore for new genes of the medium-chain dehydrogenase/reductase enzyme superfamily . Our group has recently identified a new gene, the YMR318C open reading frame, which coded for a Zn-containing NADP(H)-dependent alcohol dehydrogenase (ADHVI) . ADHVI has been purified to homogeneity from over expressing yeast cells, and found to be a homodimer of 40 kDa subunits . The enzyme showed a strict specificity for NADP(H) and high activity with a variety of long chain aliphatic and bulky substrates . Aldehydes exhibited 50-12000 times higher catalytic efficiency than the corresponding alcohols . Substrates with high k(cat)/K(m) were: pentanal, veratraldehyde and cinnamaldehyde . The ADHVI expression was strongly induced when galactose was the sole carbon source in the culture medium . Phylogenetic trees include ADHVI in the cinnamyl alcohol dehydrogenase (CADH) family . In contrast to the plant CADH, involved in lignin biosynthesis, this is not the function for ADHVI, since yeast does not synthesize lignin . ADHVI may be physiologically involved in several steps of the lignin degradation pathway, initiated by other microorganisms, in the synthesis of fusel alcohols, products derived from the aminoacidic metabolism, and in the homeostasis of NADP(H) . Disruption of ADH6 was not lethal for the yeast, under laboratory conditions.

J Environ Biol, 2002 Apr, 23(2), 169 - 73
Researches on Türkoğlu Gavur Lake in Kahramanmaraş-Turkey and some projections from past to future; Inac S et al.; Gavur Lake, takes place on the Saghk Plain in the south of Turkoglu subprovince in Kahramanmaras, was one of the most important places, that was available in the past time, south of our homeland . Gavur Lake represents the northest point of the bird immigration route that extends to Victoria Lake in Africa together with Emen and Amik Lakes dried locate on more south . Gavur Lake had been dried because of the some reasons those were being the marsh inn time by without oxygen cycle and emerging the situation of the microorganisms those were big threat for human health and to provide land in 1950's for the people of immigrants in Turkoglu . Presently, the big part of the Gavur Lake has dried . Drying activities are continuing . Whereas, Gavur Lake have all the conditions of the Ramsar Accord . With the improving of the Turkoglu reed bed the local people will earn more money by tourism than agricultural . Otherwise, the future of the Gavur Lake is going to be like Amik Lake.

J Hum Genet, 2003, 48(2), 82 - 5
Haplotype analysis of the human collectin placenta 1 (hCL-P1) gene; Ohmori H et al.; Collectins are a family of C-type lectins found in vertebrates . These proteins have four regions, a relatively short N-terminal region, a collagen-like region, an alpha-helical coiled coil, and a carbohydrate recognition domain . Collectins are involved in host defense through their ability to bind carbohydrate antigens on microorganisms . Type A scavenger receptors are classical-type scavenger receptors that also have collagen-like domains . We previously described a new scavenger receptor, collectin from placenta {collectin placenta 1 (CL-P1)} . CL-P1 is a type II membrane protein with all four regions . We found that CL-P1 can bind and phagocytize both bacteria and yeast . In addition to that, it reacts with oxidized low-density lipoprotein (LDL) but not with acetylated LDL . These results suggest that CL-P1 might play important roles in host defenses and/or atherosclerosis formation . One rational strategy to study the role of CL-P1 in these pathological conditions would be to perform a haplotype association study using human samples . As a first step for this strategy, we analyzed the haplotype structure of the CL-P1gene . By sequencing the CL-P1 gene in ten Japanese volunteers, we identified five single-nucleotide polymorphisms (SNPs) with a minor allele frequency of at least 29% . To obtain SNPs in the 5'-upstream region of the gene, we screened a total of 20 SNPs described in the database and finally picked up one SNP for the present study . Thus, a total of six SNPs, one in the 5'-upstream region, two in intron 2, one in exon 5, and two in exon 6, were used to analyze the haplotype structure of the gene, with DNAs derived from 54 individuals (108 alleles) . The analysis revealed that only two of six SNPs showed significant linkage disequilibrium ( r(2) > 0.5) with each other . This haplotype information may be useful in disease-association studies in which a contribution of the CL-P1 gene has been suspected, especially in immunological disturbance or atherosclerosis . Two SNPs in exon 6, both leading to amino acid substitutions, could be candidates for influencing disease susceptibility.

Proc Natl Acad Sci U S A, 2003 Mar 18, 100(6), 3031 - 4 Epub 2003 Feb 24.
Investigating antibody-catalyzed ozone generation by human neutrophils; Babior BM et al.; Recent studies have suggested that antibodies can catalyze the generation of previously unknown oxidants including dihydrogen trioxide (H(2)O(3)) and ozone (O(3)) from singlet oxygen ((1)O(2)(*)) and water . Given that neutrophils have the potential both to produce (1)O(2)(*) and to bind antibodies, we considered that these cells could be a biological source of O(3) . We report here further analytical evidence that antibody-coated neutrophils, after activation, produce an oxidant with the chemical signature of O(3) . This process is independent of surface antibody concentration down to 50% of the resting concentration, suggesting that surface IgG is highly efficient at intercepting the neutrophil-generated (1)O(2)(*) . Vinylbenzoic acid, an orthogonal probe for ozone detection, is oxidized by activated neutrophils to 4-carboxybenzaldehyde in a manner analogous to that obtained for its oxidation by ozone in solution . This discovery of the production of such a powerful oxidant in a biological context raises questions about not only the capacity of O(3) to kill invading microorganisms but also its role in amplification of the inflammatory response by signaling and gene activation.

Chemosphere, 2003 May, 51(5), 419 - 28
Biological oxidation of arsenite: batch reactor experiments in presence of kutnahorite and chabazite; Lievremont D et al.; Arsenic represents a threat to all living organisms due to its toxicity which depends on its speciation . This element is carcinogenic, teratogenic and is certainly one of the most important contaminants affecting millions of people around the world . Abiotic and biotic processes control its speciation and distribution in the environment . We have previously shown that a new bacterial strain named ULPAs1 performed oxidation of As(III) (1.33 mM) to As(V) in batch cultures . In order to develop new methods to remove arsenic from contaminated effluents or waste, by bacterial oxidation of As(III) to As(V) followed by its sorption, the conservation of oxidative properties of ULPAs1 was investigated when cultivated in batch reactors in the presence of two solid phases, chabazite and kutnahorite, already used as microorganisms immobilizing materials in biological remediation processes . In parallel, the retention efficiency of these solid phases toward arsenic ions and particularly arsenate was studied . Pure quartz sand was used as a reference material . Kutnahorite efficiently sorbed As(V), chabazite alone performed As(III) oxidation and pure quartz sand did not sorb arsenic at all . The arsenite oxidative properties of ULPAs1 were conserved when cultivated in the presence of quartz or chabazite.

Pol J Pathol, 2002, 53(4), 223 - 8
Immunofluorescence in situ and the serologic indices of Chlamydia pneumoniae infection in patients with an abdominal aortic aneurysm; Wolski A et al.; The aim of the study was to evaluate the frequency of Chlamydia pneumoniae infection in patients with abdominal aortic aneurysm (AAA) using selected methods . The histological specimens of aneurysm wall were evaluated, the method of immunofluorescence was used to reveal the antigen in the wall of AAA and the titers of specific antibodies of IgG, IgM and IgA classes in blood plasma were marked . Atherosclerotic changes in the aneurysm wall were found in all patients . In 20(87%) patients the C . pneumoniae antigen was seen in the wall of abdominal aneurysm using the indirect immunofluorescence method . A significant relation between the method of direct C . pneumoniae diagnosis, aneurysm symptoms and histologically detected inflammation in its wall was confirmed . Serologic markers of the chronic C . pneumoniae infection were seen in 20(87%) out of 23 patients and in 6(30%) out of 20 subjects of the control group and this difference was statistically significant . It was observed, that all patients with serologic indices of active C . pneumoniae infection, had symptomatic aneurysm . The presence of Chlamydia pneumoniae in the wall of AAA as well as the occurrence of serologic indices of the chronic infection in these patients can confirm the hypothesis of the relation between the infection with this microorganism and the development of the disease.

Indian J Exp Biol, 2002 Aug, 40(8), 874 - 81
Retinoic acid--a player that rules the game of life and death in neutrophils; Mehta K; Neutrophils are the most prevalent white blood cells in the circulation . They represent the first line of defense against invading microorganisms and have been implicated in the pathogenesis of a number of human diseases . In response to various factors, the pluripotent stem cells in bone marrow differentiate into mature neutrophils, enter the blood stream, and die within 24 hr via apoptosis . Numerous defects can occur during the process of neutrophils' differentiation that can manifest in the form of a variety of clinical disorders . Retinoids (Vitamin A and analogues), in general, and all-trans retinoic acid (tRA), in particular, play a critical role during differentiation of neutrophils . tRA can directly modulate gene expression via binding to its nuclear receptors, which in turn, can activate transcription of genes that are essential for differentiation of immature cells to neutrophils . Involvement of retinoic acid receptor in pathogenesis of acute promyelocytic leukemia (APL), reflects an important role played by this receptor in differentiation of immature myeloid cells to neutrophils . This review summarizes evidence on involvement of retinoic acid-mediated events in differentiation process of neutrophils and their subsequent apoptosis.

Biosci Biotechnol Biochem, 2002 Dec, 66(12), 2651 - 7
Old Yellow Enzyme from Candida macedoniensis catalyzes the stereospecific reduction of the C=C bond of ketoisophorone; Kataoka M et al.; Microorganisms were screened for ones that reduced 3,5,5-trimethyl-2-cyclohexene-1,4-dione (ketoisophorone; KIP), and several strains were found to produce (6R)-2,2,6-trimethylcyclohexane-1,4-dione (levodione) . The enzyme catalyzing the reduction of the C=C bond of KIP to yield (6R)-levodione was isolated from Candida macedoniensis AKU4588 . The results of primary structural analysis and its enzymatic properties suggested that the enzyme might be an Old Yellow Enzyme family protein.

Biosci Biotechnol Biochem, 2002 Dec, 66(12), 2614 - 20
Novel enzymatic method for the production of xylitol from D-arabitol by Gluconobacter oxydans; Suzuki S et al.; Microorganisms capable of producing xylitol from D-arabitol were screened for . Of the 420 strains tested, three bacteria, belonging to the genera Acetobacter and Gluconobacter, produced xylitol from D-arabitol when intact cells were used as the enzyme source . Among them, Gluconobacter oxydans ATCC 621 produced 29.2 g/l xylitol from 52.4 g/l D-arabitol after incubation for 27 h . The production of xylitol was increased by the addition of 5% (v/v) ethanol and 5 g/l D-glucose to the reaction mixture . Under these conditions, 51.4 g/l xylitol was obtained from 52.4 g/l D-arabitol, a yield of 98%, after incubation for 27 h . This conversion consisted of two successive reactions, conversion of D-arabitol to D-xylulose by a membrane-bound D-arabitol dehydrogenase, and conversion of D-xylulose to xylitol by a soluble NAD-dependent xylitol dehydrogenase . Use of disruptants of the membrane-bound alcohol dehydrogenase genes suggested that NADH was generated via NAD-dependent soluble alcohol dehydrogenase.

Biomedica, 2002 Dec, 22(4), 510 - 8
{Phenotypic diagnosis of primary immunodeficiencies in Antioquia, Colombia, 1994-2002}; Montoya CJ et al.; Recurrent infections are a frequent cause of medical visits . They can be due to a heterogeneous group of dysfunctions that increase the susceptibility to pathogenic and opportunistic microorganisms, such as immunological deficiencies . To define an opportune rational treatment and to guide the molecular diagnosis of primary immunodeficiency diseases, we establish a program for the phenotypic diagnosis of these illnesses in Antioquia, Colombia, including clinical and laboratory evaluations of patients who present recurrent infections with abnormal evolution . Between August 1, 1994 and July 31, 2002, phenotypic diagnosis of primary immunodeficiency was made in 98 patients . Similar to data reported in the literature, antibody deficiencies were the most frequent (40.8%), followed by combined deficiencies (21.4%) . This phenotypic characterization has allowed for appropriate treatments for each patient and, in some cases, functional and molecular studies that can lead to a definite molecular diagnosis.

J Basic Microbiol, 2003, 43(1), 68 - 74
The influence of co-cultivation on expression of the antifungal protein in Aspergillus giganteus; Meyer V et al.; The afp gene of Aspergillus giganteus encodes a small, highly basic polypeptide with antifungal activity, named Antifungal Protein (AFP) . The protein is secreted by the mould and inhibits the growth of various filamentous fungi . In this paper we report that co-cultivation of A . giganteus with various microorganisms alters afp expression . It was found that co-cultivation modulates afp expression on the level of transcription, using a reporter system based on the beta-glucuronidase gene . The presence of Fusarium oxysporum triggered afp transcription whereas dual cultures of A . giganteus and A . niger resulted in suppression of afp transcription . Growth tests performed with several carbon and nitrogen sources, revealed that the influence of co-cultivation is strongly dependent on the medium composition.

Biochimie, 2002 Nov, 84(11), 1173 - 9
Quality control of biotechnology-derived vaccines: technical and regulatory considerations; Fuchs F; Vaccines for human use have been produced for decades using classical manufacturing methods including culture of viruses and bacteria followed by various concentration-, inactivation-, detoxification-, conjugation production processes . Availability of techniques for molecular biology and for the complete chemical synthesis of genes provides prospects of genetic engineering of microorganisms so as to generate novel biotechnological/biological-derived vaccines . The potential large-scale availability of biotechnology-derived vaccines makes feasible their evaluation in the prevention and/or treatment of various infectious, chronic, degenerative and cancer human diseases . There are potential safety concerns that arise from the novel manufacturing processes and from the complex structural and biological characteristics of the products . These products have distinguishing characteristics to which consideration should be given in a well-defined quality control testing programme . The evaluation of their quality, safety, efficacy and stability necessitate complex analytical methods and appropriate physicochemical, biochemical and immunochemical methods for the analysis of the molecular entity . A flexible approach to the control of these novel products is being developed by regulatory authorities so that recommendations can be modified in the light of experience of research and development in vaccinology, production and use of biotechnology products and with the further development of new technologies.

Int J Food Microbiol, 2003 May 15, 82(3), 233 - 43
A hybrid Bayesian-neural network approach for probabilistic modeling of bacterial growth/no-growth interface; Hajmeer MN et al.; A hybrid probabilistic modeling approach that integrates artificial neural networks (ANNs) with statistical Bayesian conditional probability estimation is proposed . The suggested approach benefits from the power of ANNs as highly flexible nonlinear mapping paradigms, and the Bayes' theorem for computing probabilities of bacterial growth with the aid of Parzen's probability distribution function estimators derived for growth and no-growth (G/NG) states . The proposed modeling approach produces models that can predict the probability of growth of targeted microorganism as affected by a set of parameters pertaining to extrinsic factors and operating conditions . The models also can be used to define the probabilistic boundary (interface) between growth and no-growth, and as such can define and predict the values of critical parameters required to keep a desired pre-specified bacterial growth risk in check . A modular system incorporating the various computational modules was constructed to illustrate the application of the hybrid approach to the probabilistic modeling of growth of pathogenic Escherichia coli strain as affected by temperature and water activity . The proposed approach was compared to other techniques including the traditional linear and nonlinear logistic regression . Results indicated that the hybrid approach outperforms the other approaches in its accuracy as well as flexibility to extract the implicit interrelationships between the various parameters . Advantages and limitations of the approach were also discussed and compared to those of other techniques.

Astrobiology, 2002 Winter, 2(4), 383 - 402
Long-term manipulations of intact microbial mat communities in a greenhouse collaboratory: simulating earth's present and past field environments; Bebout BM et al.; Photosynthetic microbial mat communities were obtained from marine hypersaline saltern ponds, maintained in a greenhouse facility, and examined for the effects of salinity variations . Because these microbial mats are considered to be useful analogs of ancient marine communities, they offer insights about evolutionary events during the >3 billion year time interval wherein mats co-evolved with Earth's lithosphere and atmosphere . Although photosynthetic mats can be highly dynamic and exhibit extremely high activity, the mats in the present study have been maintained for >1 year with relatively minor changes . The major groups of microorganisms, as assayed using microscopic, genetic, and biomarker methodologies, are essentially the same as those in the original field samples . Field and greenhouse mats were similar with respect to rates of exchange of oxygen and dissolved inorganic carbon across the mat-water interface, both during the day and at night . Field and greenhouse mats exhibited similar rates of efflux of methane and hydrogen . Manipulations of salinity in the water overlying the mats produced changes in the community that strongly resemble those observed in the field . A collaboratory testbed and an array of automated features are being developed to support remote scientific experimentation with the assistance of intelligent software agents . This facility will permit teams of investigators the opportunity to explore ancient environmental conditions that are rare or absent today but that might have influenced the early evolution of these photosynthetic ecosystems.

Surg Infect (Larchmt), 2001 Fall, 2(3), 241 - 5
Handwashing compliance depends on professional status; Lipsett PA et al.; BACKGROUND: Nosocomial infections can be transmitted from microorganisms on the hands of health care workers to patients . Handwashing (HW) has a proven benefit in preventing transmission of infection, yet compliance with handwashing, especially in intensive care units, ranges between 28% and 74% . METHODS: To determine if HW behavior varies as a function of health care professional status and patient interaction, we conducted an observational study of a surgical intermediate care unit in a large university teaching hospital . HW compliance was observed among all health care workers (HCW): physicians (MD; N = 46), nurses (RN; N = 295), and nursing support personnel (NSP; N = 93) . Over an 8-week period, unidentified, trained observers documented all HCW interactions in 1-h random blocks . HW opportunities were classified into low and high risk of pathogen acquisition and transmission . RESULTS: A total of 493 HW opportunities were observed, of which 434 involved MD, RN, and NSP . Two hundred and sixty-one low-risk (MD 35, RN 171, NSP 55) and 173 (MD 11, RN 124, NSP 38) high-risk interactions were observed . Overall HW rates were low (44%) . Significant differences existed among HCW, with MDs being the least likely to wash (15% versus RN 50%, NSP 37%, p < 0.01) . In adjusting for high-risk situations, MDs (odds ratio {OR} 5.58, 95% CI 2.49-12.54; NSP, OR 1.73, 95% CI 1.13-2.64; RN, OR 0.98, 95% CI 0.77-1.23) were significantly less likely to perform HW when compared to RNs . Nursing groups were significantly less likely to wash in low-risk versus high-risk situations (MD 9.2% versus 17.1%; RN 69.4% versus 39.6%; NSP 85% versus 23.3%), suggesting individual discrimination of the importance of HW . Although nurses were less likely to wash in high-risk situations compared to NSP, the overall number of opportunities was greater, suggesting that improvement in HW to the level of NSP could have a major impact on infection transmission . CONCLUSION: Significant opportunities exist for quality improvement, novel educational strategies, and assessment of reasons why MDs and, to a lesser extent, RNs fail to follow simple HW practices.

J Periodontol, 2003 Jan, 74(1), 66 - 75
Neutrophil-mediated tissue injury in periodontal disease pathogenesis: findings from localized aggressive periodontitis; Kantarci A et al.; Neutrophils play a major role in the host response against invading periodontopathogenic microorganisms . Localized aggressive periodontitis (LAgP) is associated with various functional abnormalities of neutrophils . Based on the recent findings, LAgP neutrophils are not "hypofunctional" or "deficient." They are "hyperfunctional," and their amplified activity is responsible for the tissue destruction in periodontal disease . Several signal transduction abnormalities are associated with elevated neutrophil function in LAgP . There is a strong correlation between defective chemotaxis and decreased intracellular Ca2+ levels; total calcium-dependent protein Kinase C (PKC) activity of neutrophils is significantly lower than healthy subjects; and there is a marked increase in diacylglycerol (DAG) accompanied by a pronounced decrease in DAG kinase activity . In a separate set of experiments on the involvement of the inducible cyclooxygenase isoform (COX-2) and the role of novel lipid mediators in the pathogenesis of periodontal disease, crevicular fluid samples from LAgP patients were found to contain prostaglandin E2 (PGE2) and 5-LO-derived products, leukotriene B4 (LTB4), and the biosynthesis interaction product, lipoxin LXA4 . Neutrophils from peripheral blood of LAgP patients, but not from healthy volunteers, also generated LXA4, suggesting that this immunomodulatory molecule may have a role in periodontal disease . Lipoxin generation and its relationship to PGE2 and LTB4 can be visualized as an important marker for the pathogenesis of periodontal disease . Thus, major advances in our understanding of the role of the neutrophil in host defense against periodontal organisms have been made through studies of LAgP . LAgP is used as an example of a severe periodontal disease that is related to abnormal neutrophil function . In this model, it appears that a hyperresponsiveness of the neutrophil, due to cell priming/predisposition, results in enhanced tissue damage.

Infection, 2003 Jan, 31(1), 31 - 7
Prolonged application of closed in-line suction catheters increases microbial colonization of the lower respiratory tract and bacterial growth on catheter surface; Freytag CC et al.; BACKGROUND: Microorganisms become attracted to polymer surfaces for a number of reasons including positive charge of biomaterial or concentration of nutrients on the biomaterial surface . Many bacteria additionally possess specific receptors for the interaction with extracellular host protein components that adhere on the biomaterial surface . Several authors suggest that application of closed in-line polypropylene suction catheters (CISC) in intubated patients for more than 24 h is safe and can reduce the costs associated with mechanical ventilation . Therefore, we evaluated the possible role of prolonged application of CISC to cause enhanced colonization of both the biomaterial and the lower respiratory tract . MATERIALS AND METHODS: The prospective, randomized study included 23 mechanically ventilated patients . The CISC tips, adjacent segments and tracheobronchial aspirates of each patient were examined for microbial growth . RESULTS: Application for 72 h significantly enhanced the microbial growth on the CISC tips and on the adjacent catheter segment . Usage for 3 days led to a significant increase in colonization in the lower respiratory tract . CONCLUSION: Normal saline instillation in conjunction with endotracheal suctioning may lead to a dispersion of microorganisms into the lower respiratory tract . More effective self-cleaning mechanisms are necessary to decontaminate the CISC surface after suctioning.

Am J Respir Crit Care Med, 2003 Feb 15, 167(4), 593 - 8
Pulmonary matrix metalloproteinase excess in hospital-acquired pneumonia; Hartog CM et al.; In hospital-acquired pneumonia, extracellular matrix destruction is common and may be caused by excessive activity of matrix metalloproteinases (MMPs) . Thirty patients with hospital-acquired pneumonia and 16 control subjects were studied . We evaluated the concentrations of MMP-8, MMP-9, and tissue inhibitor of metalloproteinase-1 in mini-bronchoalveolar lavage fluid (mini-BALF) and blood using zymography and specific immunoassays . In patients with hospital-acquired pneumonia concentrations of MMP-8 and MMP-9 in mini-BALF were increased 10-fold, whereas their specific inhibitor tissue inhibitor of metalloproteinase-1 was not concomitantly increased . In 80% of patients with pneumonia, but in none of the control subjects, the active form of MMP-9 was detected by zymography . Zymography furthermore showed the banding pattern of neutrophil-derived MMP-9, indicating that neutrophils were the main source of MMP-9 . Comparison of neutrophils from blood and mini-BALF showed higher basal release of MMPs by pulmonary neutrophils . Stimulation analysis indicated that pulmonary neutrophils were already maximally activated . In patients with detection of potentially pathogenic microorganisms, concentrations of MMPs were fivefold increased compared with patients with negative cultures . Furthermore, MMP-levels were related to clinical severity . These are the first data suggesting that neutrophil-derived MMPs are increased in hospital-acquired pneumonia in association to the detection of causative microorganisms and clinical severity.

J Appl Microbiol, 2003, 94(3), 475 - 82
Temperature effect on Fusarium oxysporum f.sp . melonis survival during horticultural waste composting; Suarez-Estrella F et al.; AIMS: The aim of this work was to study the effect of high temperatures generated during composting process, on the phytopathogen fungus Fusarium oxysporum f.sp . melonis . This investigation was achieved by both in vivo (semipilot-scale composting of horticultural wastes) and in vitro (lab-scale thermal treatments) assays . METHODS AND RESULTS: Vegetable residues infected with F . oxysporum f.sp . melonis were included in compost piles . Studies were conducted in several compost windrows subjected to different treatments . Results showed an effective suppression of persistence and infective capacity, as this process caused complete fungal elimination after 2-3 days of composting . In order to confirm the effect of high temperature during this process, in vitro experiments were carried out . Temperature values of 45, 55 and 65 degrees C were tested . All three treatments caused the elimination of fungal persistence . Treatment at 65 degrees C was especially effective, whereas 45 degrees C eliminated fungal persistence only after 10 days . CONCLUSIONS: The composting process is an excellent alternative for the management of plant wastes after harvesting, as this procedure is able to suppress infective capacity of several harmful phytopathogens such as F . oxysporum f.sp . melonis . SIGNIFICANCE AND IMPACT OF THE STUDY: Fusarium oxysporum f.sp . melonis is a plant pathogen fungus specially important in the province of Almeria (south-east Spain), where intensive greenhouse horticulture is very extended . High temperatures reached during composting of horticultural plant wastes ensure the elimination of phytopathogen microorganisms such as F . oxysporum f.sp . melonis from vegetable material, providing an adequate hygienic quality in composts obtained.

Br J Dermatol, 2003 Feb, 148(2), 224 - 8
Pattern analysis of human cutaneous mast cell populations by total body surface mapping; Weber A et al.; BACKGROUND: Mast cells (MCs), critical effector cells in allergic inflammation and innate immunity to bacteria, are located in large numbers in tissues that interface the external environment, including the skin . However, little is known about the distribution and numbers of human skin MCs . OBJECTIVES: To assess the influence of age, sex and skin region on size and spatial distribution of MC populations in normal human skin . METHODS: Biopsies of healthy skin were obtained from 150 male and female individuals (age range 10-86 years) . MCs were quantified and mapped planimetrically by histomorphometry in 15 anatomical sites (abdomen, thorax, lower and upper back, lower and upper arm, lower and upper leg, foot, hand, chin, nose, cheek, forehead, back of head) . RESULTS: No differences in skin MC numbers or distribution were found when comparing skin obtained from male or female and from young or old individuals . At all skin sites, regardless of age or sex, MC numbers were highest in the most superficial skin layers where up to 10-fold more MCs were found as compared with the subcutis, which consistently contained the lowest numbers of MCs . Interestingly, MC numbers were highest at peripheral skin sites (maximum: chin and nose) and lowest at central skin sites (minimum: abdomen) . Thus, healthy human skin exhibits a proximal/distal and a central/peripheral MC gradient and 'skin MC numbers' may vary by a factor of more than 20, depending on the skin layer and skin site analysed . CONCLUSIONS: These findings support the recently identified role of MCs in the elicitation of protective immune responses against infectious microorganisms . One possible explanation for these unexpected findings is that skin site/layer-specific factors (e.g . the density of nerves or vessels, exposure to ultraviolet irradiation, frequency of minimal trauma) are involved in the regulation of skin MC numbers and distribution.

Environ Microbiol, 2003 Mar, 5(3), 212 - 6
Molecular diversity among marine picophytoplankton as revealed by psbA analyses; Zeidner G et al.; Photosynthetic microorganisms play a crucial role in the marine environment . In vast areas of the oceans, marine primary productivity is performed by cells smaller than 2-3 micro m (picoplankton) . Here, we report on molecular analyses of the conserved photosynthetic psbA gene (coding for protein D1 of photosystem II reaction centre) as a diversity indicator of naturally occurring marine oxygenic picophytoplankton . The psbA genes proved to be good indicators of the presence of a wide variety of photosynthetic marine microbial groups, including new cyanobacterial groups and eukaryotic algae (prasinophytes) . Furthermore, using environmental bacterial artificial chromosome (BAC) libraries, we were able to correlate psbA genes with small subunit rRNAs and, therefore, to confirm their phylogenetic affiliation.

Eur J Clin Invest, 2003 Feb, 33(2), 169 - 75
Pneumococcal immune adherence to human erythrocytes; Hament JM et al.; BACKGROUND: Human red blood cells bind various C3b-coated microorganisms via their C3b/CR1 receptor, a phenomenon referred to as immune adherence . The aim of the present study was to measure pneumococcal adherence to human red blood cells by flow cytometry and to study kinetic aspects of this binding . MATERIAL AND METHODS: We quantified pneumococcal adherence to human erythrocytes by FACS analysis and tested the involvement of antibodies and complement activation in this process . RESULTS: Pneumococci are able to bind to human red blood cells in the presence of human serum . Coating with C3b/C4b appeared obligatory for pneumococcal adherence to red blood cells . The ligand on erythrocytes was confirmed to be complement receptor 1 . Kinetic studies showed that innate (mannose-binding lectin) and specific immune factors (IgG antibodies) contributed to the binding of C3b-coated pneumococci to human erythrocytes . After initial binding, serum-derived factor I was found to induce bacterial detachment from the erythrocyte . CONCLUSIONS: Pneumococci are able to adhere to red blood cells . Both the classical and lectin complement pathways are important for optimal C3b-coating of pneumococci for immune adherence . Bound pneumococci are detached from red blood cells by factor I . These findings are in line with the hypothesis of immune adherence in which human erythrocytes are able to bind pneumococci and target the bacteria to the reticulo-endothelial system in the spleen.

Rev Environ Contam Toxicol, 2000, 164, 1 - 26
Metalaxyl: persistence, degradation, metabolism, and analytical methods; Sukul P et al.; Metalaxyl is a systemic fungicide used to control plant diseases caused by Oomycete fungi . Its formulations include granules, wettable powders, dusts, and emulsifiable concentrates . Application may be by foliar or soil incorporation, surface spraying (broadcast or band), drenching, and seed treatment . Metalaxyl registered products either contain metalaxyl as the sole active ingredient or are combined with other active ingredients (e.g., captan, mancozeb, copper compounds, carboxin) . Due to its broad-spectrum activity, metalaxyl is used world-wide on a variety of fruit and vegetable crops . Its effectiveness results from inhibition of uridine incorporation into RNA and specific inhibition of RNA polymerase-1 . Metalaxyl has both curative and systemic properties . Its mammalian toxicity is classified as EPA toxicity class III and it is also relatively non-toxic to most nontarget arthropod and vertebrate species . Adequate analytical methods of TLC, GLC, HPLC, MS, and other techniques are available for identification and determination of metalaxyl residues and its metabolites . Available laboratory and field studies indicate that metalaxyl is stable to hydrolysis under normal environmental pH values, It is also photolytically stable in water and soil when exposed to natural sunlight . Its tolerance to a wide range of pH, light, and temperature leads to its continued use in agriculture . Metalaxyl is photodecomposed in UV light, and photoproducts are formed by rearrangement of the N-acyl group to the aromatic ring, demethoxylation, N-deacylation, and elimination of the methoxycarbonyl group from the molecule . Photosensitizers such as humic acid, TiO2, H2O2, acetone, and riboflavin accelerate its photodecomposition . Information is provided on the fate of metalaxyl in plant, soil, water, and animals . Major metabolic routes include hydrolysis of the methyl ester and methyl ether oxidation of the ring-methyl groups . The latter are precursors of conjugates in plants and animals . In soils the most relevant metabolite is the metalaxyl acid, which is formed predominantly by soil microorganisms . Plant uptake, microbial degradation, photodecomposition, and leaching are the major route of metalaxyl dissipation . It has a tendency to migrate to deeper soil horizons with a potential to contaminate groundwater, particularly in soils with low organic matter and clay content . Therefore, precautions should be taken for the continuous application of metalaxyl to crops . If use of metalaxyl is greately increased, the risk of occurrence in groundwater must be reassessed, as by monitoring studies in the most vulnerable areas in main use regions . The R-isomer of metalaxyl (mefenoxam) has recently been registered as the only active compound . Therefore, quantitative studies on the fate of this specific isomer are needed, including appropriate analytical methods . As the use rates of mefenoxam are approximately one-half those recommended for metalaxyl and mefenoxam dissipates more rapidly, concerns for mefenoxam reaching groundwater are even less justified.

Indian J Exp Biol, 2002 Sep, 40(9), 1083 - 6
Production of glycolipids containing biosurfactant by Pseudomonas species; Ellaiah P et al.; Microorganisms, that degrade hydrocarbon were isolated and screened for their biosurfactant activity . A total of 68 strains were isolated and tested for their glycolipid activity of which 4 isolates showed good glycolipid activity . Isolate K10 gave the maximum biosurfactant production in medium A (containing kerosene as a sole carbon source) as compared to medium B (containing glucose as a sole carbon source) . Characterization of isolate K10 showed that it belongs to Pseudomonas species.

FEMS Microbiol Lett, 2003 Jan 28, 218(2), 223 - 30
Cloning and characterization of a gene cluster for geldanamycin production in Streptomyces hygroscopicus NRRL 3602; Rascher A et al.; We illustrate the use of a PCR-based method by which the genomic DNA of a microorganism can be rapidly queried for the presence of type I modular polyketide synthase genes to clone and characterize, by sequence analysis and gene disruption, a major portion of the geldanamycin production gene cluster from Streptomyces hygroscopicus var . geldanus NRRL 3602.

Dev Cell, 2003 Feb, 4(2), 146 - 8
Neutrophils with a license to kill: permeabilized, not stirred; Finkel T; The neutrophil responds to invading microorganisms in part by assembling the NADPH oxidase complex and producing superoxide radicals . Relatively little is known about the intracellular assembly or activation of the oxidase, but Brown et al . in the January issue of Molecular Cell provide a useful strategy involving permeabilized neutrophils to tackle this question.

Anal Chem, 2003 Feb 1, 75(3), 694 - 7
Use of bioactive glass slides for matrix-assisted laser desorption/ionization analysis: application to microorganisms; Afonso C et al.; Glass slides are widely used in high-throughput analysis and are available commercially with surfaces activated, etched, and channeled . Thin glass microscope slides are shown here to be suitable sample supports for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry . As a demonstration, lectins immobilized on glass slides with activated surfaces are used to concentrate and purify agglutinated Bacillis spores . It is expected that such slides will provide a rapid, inexpensive way to evaluate and implement new strategies involving MALDI MS readout.

Biotechnol Bioeng, 2003 Apr 20, 82(2), 180 - 9
A light distribution model for an internally radiating photobioreactor; Suh IS et al.; Analysis of light energy distribution in culture is important for maximizing the growth efficiency of photosynthetic cells and the productivity of a photobioreactor . To characterize the irradiance conditions in a photobioreactor, we developed a light distribution model for a single-radiator system and then extended the model to multiple radiators using the concept of parallel translation . Mathematical expressions for the local light intensity and the average light intensity were derived for a cylindrical photobioreactor with multiple internal radiators . The proposed model was used to predict the irradiance levels inside an internally radiating photobioreactor using Synechococcus sp . PCC 6301 as a model photosynthetic microorganism . The effects of cell density and radiator number were interpreted through photographic and model simulation studies . The predicted light intensity values were found to be very close to those obtained experimentally, which suggests that the proposed model is capable of accurately interpreting the local light energy profiles inside the photobioreactor system . Due to the simplicity and flexibility of the proposed model, it was also possible to predict the light conditions in other complex photobioreactors, including optical-fiber and pond-type photobioreactors .

Adv Skin Wound Care, 2003 Jan-Feb, 16(1), 12 - 23; quiz 24-5
The clinical relevance of microbiology in acute and chronic wounds; McGuckin M et al.; PURPOSE: To improve clinical practice and the quality of patient care by providing a learning opportunity that enhances the participant's understanding of how wound microbiology affects healing . TARGET AUDIENCE: This CME/CE activity is intended for physicians and nurses with an interest in interpreting the role of microorganisms in wound healing . OBJECTIVES: At the conclusion of this course, participants should be able to: 1 . Identify the microbiology of acute and chronic wounds, risk factors for infection, and advantages and disadvantages of wound culturing . 2 . Identify methods of debridement and wounds for which they are appropriate . 3 . Identify systemic antibiotic treatment options for acute and chronic wound infections.

Lett Appl Microbiol, 2003, 36(3), 177 - 81
Use of CAS-agar plate modified to study the effect of different variables on the siderophore production by Aspergillus; Machuca A et al.; AIMS: To evaluate the suitability of chrome azurol S (CAS) agar plate assay as a quantitative methodology for siderophore production . METHODS AND RESULTS: Aspergillus species (A . flavus, A . niger, A . tamarii) were inoculated in the CAS-agar plates and the siderophores production was determined and expressed as CAS-reaction rate (mm per day) . All the species showed positive CAS reaction with different rates depending on culture conditions and A . flavus showed the highest CAS-reaction rate . The siderophore production in solid medium expressed as CAS-reaction rate was correlated with siderophore production in liquid medium . CONCLUSIONS: The use of CAS-agar plate assay was modified and the evaluation of CAS reaction in mm per day made it possible to study and quantify the effect of several variables on the siderophore production by Aspergillus fungi . SIGNIFICANCE AND IMPACT OF THE STUDY: We describe the CAS-agar plate assay as a quantitative methodology, which make it possible to select and evaluate the siderophore production by several microorganisms (fungi and bacteria) according to different culture conditions.

Mol Microbiol, 2003 Feb, 47(4), 879 - 89
Microarray expression profiling: capturing a genome-wide portrait of the transcriptome; Conway T et al.; The bacterial transcriptome is a dynamic entity that reflects the organism's immediate, ongoing and genome-wide response to its environment . Microarray expression profiling provides a comprehensive portrait of the transcriptional world enabling us to view the organism as a 'system' that is more than the sum of its parts . The vigilance of microorganisms to environmental change, the alacrity of the transcriptional response, the short half-life of bacterial mRNA and the genome-scale nature of the investigation collectively explain the power of this method . These same features pose the most significant experimental design and execution issues which, unless surmounted, predictably generate a distorted image of the transcriptome . Conversely, the expression profile of a properly conceived and conducted microarray experiment can be used for hypothesis testing: disclosure of the metabolic and biosynthetic pathways that underlie adaptation of the organism to chang-ing conditions of growth; the identification of co-ordinately regulated genes; the regulatory circuits and signal transduction systems that mediate the adaptive response; and temporal features of developmental programmes . The study of bacterial pathogenesis by microarray expression profiling poses special challenges and opportunities . Although the technical hurdles are many, obtaining expression profiles of an organism growing in tissue will probably reveal strategies for growth and survival in the host's microenvironment . Identifying these colonization strategies and their cognate expression patterns involves a 'deconstruction' process that combines bioinformatics analysis and in vitro DNA array experimentation.

Cell Microbiol, 2003 Feb, 5(2), 75 - 83
In the belly of the beast: subversion of macrophage proinflammatory signalling cascades during Toxoplasma gondii infection; Denkers EY et al.; Macrophages (MO) are used as the intracellular niche by several bacterial and protozoan microorganisms . Such microbial pathogens adopt diverse strategies to avoid MO microbicidal effects . Recent insights into the Toxoplasma gondii-MO interaction reveal novel ways that intracellular parasites subvert MO function . In contrast to some microbial pathogens, Toxoplasma infection is not silent but induces rapid activation of transcription factors such as STAT-1 and NFkappaB . However, the parasite blocks nuclear translocation of both factors, and MO cannot produce IL-12 or TNF-alpha when subsequently triggered with lipopolysaccharide . The nuclear import blockade is lifted 24 h after infection, but cells remain actively suppressed in TNF-alpha production . Nevertheless, IL-12 synthesis is initiated at this later time point . Toxoplasma gondii-induced production of this cytokine occurs through both MyD88- and CCR5-dependent pathways . The balance of cytokine subversion and stimulation during infection probably results from the parasite's need to simultaneously avoid immune elimination and trigger immunity to prevent host death.

Contemp Top Lab Anim Sci, 2003 Jan, 42(1), 52 - 6
Coinfection of laboratory rats with Mycoplasma pulmonis and Chlamydia pneumoniae; Damy SB et al.; Routine examinations of conventional outbred Wistar rats in our laboratory showed increased serum levels of alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and urea . Electron microscopy and specific reactions showed C . pneumoniae and M . pulmonis in lung, liver, spleen, heart, and kidney sections . We could not exclude the fact that other infectious microorganisms detected through routine health surveillance affected the Wistar rat colony; however, we have not identified any of those microorganisms by electron microscopy of the organs listed . Natural coinfection of C . pneumoniae and M . pulmonis can occur in laboratory rats and is associated with histopathological and functional compromise of many organs . Further studies comparing different conventional animals and specific pathogen-free animals are necessary to better understand the present findings and to define whether coinfection influences the results of experimental studies with rats.

Adv Space Res, 2003, 31(1), 195 - 9
Recycling efficiencies of C, H, O, N, S, and P elements in a Biological Life Support System based on microorganisms and higher plants; Gros JB et al.; MELiSSA is a microorganism based artificial ecosystem conceived as a tool for understanding the behavior of ecosystems and developing the technology for future Manned Space Missions . MELiSSA is composed of four compartments colonized by the microorganisms required by the function of this ecosystem : breakdown of waste produced by men, regeneration of atmosphere and biosynthesis of edible biomass . This paper reports the mass balance description of a Biological Life Support System composed of the MELiSSA loop and of a Higher Plant Compartment working in parallel with the photosynthetic Spirulina compartment producing edible biomass . The recycling efficiencies of the system are determined and compared for various working conditions of the MELiSSA loop with or without the HPC . c2002 Published by Elsevier Science Ltd on behalf of COSPAR.

Eur J Med Res, 2003 Jan 28, 8(1), 25 - 32
Effects of selected immunouppressive drugs on prostaglandin release, protein synthesis and cell proliferation in human gingival fibroblasts and on the growth of plaque bacteria; Pistorius A et al.; BACKGROUND: Immunosuppressants play an essential role in transplantation therapy . In view of the side effects, e.g . gingival overgrowth, the present in vitro study was performed in order to investigate the effect of selected immunosuppressants on metabolic activities of gingival fibroblasts . Furthermore, the effect on the growth of six oral microorganisms was investigated . METHODS: Human gingival fibroblasts were incubated in the presence of azathioprine (Aza), cyclosporin A (CsA), tacrolimus (Tac) or mycophenolatmofetil (Myc) . PGE subset 2 release was determined by means of a specific competitive enzyme immunoassay, using monoclonal antibodies specific for PGE subset 2 (clone E2R1) . The protein content was measured spectrophotometrically . A redox indicator system was employed to assess the proliferation activity . In an additional trial the growth of six strains of oral bacteria (A . viscosus T14V, S . oralis H1, S . mutans 10449, C . gingivalis DR2001, A . actinomycetemcomitans Y4, and M . micros 33270) in the presence of the immunosuppressants was measured . RESULTS: In comparison with the controls, the PGE subset 2 release was increased by 39.3% following incubation with Aza, and by 77.0% with CsA . The protein concentrations (1 g immunosuppressant / ml medium) were reduced by 26.0% for Aza and 17.0% for Myc . Furthermore, a drug-dependent inhibition in the cell proliferation rate was noted after an incubation period of 6 hours (Aza 70.7%, CsA 78.2%, Myc 69.8%, Tac 64.0%) . The most pronounced growth-inhibiting effects were observed for CsA at values ranging from 21.0% (S . mutans 10449) to 48.6% (A . viscosus T14V) growth inhibition . CONCLUSIONS: The present study with common immunsuppresants demonstrated both a medication- and dose-dependent alteration in the metabolic activity of gingival fibroblasts . Furthermore, growth-inhibitory effects on the selected bacterial strains could be observed.

J Neuropathol Exp Neurol, 2003 Feb, 62(2), 154 - 61
The microvasculature of the cerebral white matter: arteries of the subcortical white matter; Nonaka H et al.; The microvascular architecture of the human cerebral subcortical white matter was studied . Most of the subcortical arteries ran straight through the cortex, but upon entering the white matter, they began to coil, loop, and spiral . Vascular stains showed wide spaces between the adventitial sheaths and blood vessels . The blood vessels coiled, looped, and spiraled within these wide adventitial spaces . This phenomenon was observed in the brains from persons ranging from the first to ninth decades of life and there were no statistically significant age-related correlations . Furthermore, there was no evidence of a reduction in the volume of white matter after fixation . Therefore, the observed tortuosity does not appear to be the result of shrinkage of brain tissue following fixation . While the mechanisms responsible for the subcortical arteries circuitry remain undetermined, this coiling architecture may serve as a trap for tumor cells and microorganisms passing through the blood stream, suggesting that these coiling arterial blood vessels may play a significant role in the pathogenesis of tumor metastasis and the brain abscess that frequently occurs in the gray-white matter junction.

Adv Space Res, 2003, 31(1), 97 - 102
Sterilisation properties of the Mars surface and atmospheric environment; Moreau D et al.; The radiative and chemical conditions at the surface and in the lower Martian atmosphere are computed at various latitudes and seasons combining a 2D photochemical model and radiation simulations . In most situations, the solar UV B and C radiations reach the surface however, suspended dust and, in polar cases, ozone can constitute an effective UV shield . The daytime and night time concentrations of the sterilizing oxidants: OH, H2O2 and O3 are determined, as well as the concentration of the substances which could influence the metabolism of microorganisms . The possible habitats of a remaining Mar's life as well as the possibilities of contamination by resistant earth life forms will be described . c2002 COSPAR . Published by Elsevier Science Ltd . All rights reserved.

Adv Space Res, 2003, 31(1), 87 - 95
Critical issues in connection with human missions to Mars: protection of and from the Martian environment; Horneck G et al.; Human missions to Mars are planned to happen within this century . Activities associated therewith will interact with the environment of Mars in two reciprocal ways: (i) the mission needs to be protected from the natural environmental elements that can be harmful to human health, the equipment or to their operations; (ii) the specific natural environment of Mars should be protected so that it retains its value for scientific and other purposes . The following environmental elements need to be considered in order to protect humans and the equipment on the planetary surface: (i) cosmic ionizing radiation, (ii) solar particle events; (iii) solar ultraviolet radiation; (iv) reduced gravity; (v) thin atmosphere; (vi) extremes in temperatures and their fluctuations; and (vii) surface dust . In order to protect the planetary environment, the requirements for planetary protection as adopted by COSPAR for lander missions need to be revised in view of human presence on the planet . Landers carrying equipment for exobiological investigations require special consideration to reduce contamination by terrestrial microorganisms and organic matter to the greatest feasible extent . Records of human activities on the planet's surface should be maintained in sufficient detail that future scientific experimenters can determine whether environmental modifications have resulted from explorations . c2002 COSPAR . Published by Elsevier Science Ltd . All rights reserved.

Bioresour Technol, 2003 Jun, 88(2), 137 - 42
Performance of a glucose fed periodic anaerobic baffled reactor under increasing organic loading conditions: 2 . Model prediction; Stamatelatou K et al.; A model was developed for the anaerobic digestion of a glucose-based medium in an innovative high-rate reactor, the periodic anaerobic baffled reactor (PABR) . The model considers each PABR compartment as two variable volume interacting sections, of constant total volume, one with high solids and one with low solids concentration, with the gas and liquid flows influencing the material flows between the two sections . For the simulation of glucose degradation, the biomass was divided into acidogenic, acetogenic and methanogenic groups of microorganisms . The kinetic part of the model accounted for possible inhibition of acidogenesis, acetogenesis and methanogenesis by volatile fatty acids . The model succeeded in predicting the reactor performance upon step increases in the organic loading rate.

Adv Space Res, 2002, 30(6), 1539 - 45
Survival of microorganisms in space protected by meteorite material: results of the experiment 'EXOBIOLOGIE' of the PERSEUS mission; Rettberg P et al.; During the early evolution of life on Earth, before the formation of a protective ozone layer in the atmosphere, high intensities of solar UV radiation of short wavelengths could reach the surface of the Earth . Today the full spectrum of solar UV radiation is only experienced in space, where other important space parameters influence survival and genetic stability additionally, like vacuum, cosmic radiation, temperature extremes, microgravity . To reach a better understanding of the processes leading to the origin, evolution and distribution of life we have performed space experiments with microorganisms . The ability of resistant life forms like bacterial spores to survive high doses of extraterrestrial solar UV alone or in combination with other space parameters, e.g . vacuum, was investigated . Extraterrestrial solar UV was found to have a thousand times higher biological effectiveness than UV radiation filtered by stratospheric ozone concentrations found today on Earth . The protective effects of anorganic substances like artificial or real meteorites were determined on the MIR station . In the experiment EXOBIOLOGIE of the French PERSEUS mission (1999) it was found that very thin layers of anorganic material did not protect spores against the deleterious effects of energy-rich UV radiation in space to the expected amount, but that layers of UV radiation inactivated spores serve as a UV-shield by themselves, so that a hypothetical interplanetary transfer of life by the transport of microorganisms inside rocks through the solar system cannot be excluded, but requires the shielding of a substantial mass of anorganic substances . c2002 COSPAR . Published by Elsevier Science Ltd . All rights reserved.

Trends Biotechnol, 2003 Feb, 21(2), 82 - 8
Global organellar proteomics; Taylor SW et al.; Cataloging the proteomes of single-celled microorganisms, cells, biological fluids, tissue and whole organisms is being undertaken at a rapid pace as advances are made in protein and peptide separation, detection and identification . For metazoans, subcellular organelles represent attractive targets for global proteome analysis because they represent discrete functional units, their complexity in protein composition is reduced relative to whole cells and, when abundant cytoskeletal proteins are removed, lower abundance proteins specific to the organelle are revealed . Here, we review recent literature on the global analysis of subcellular organelles and briefly discuss how that information is being used to elucidate basic biological processes that range from cellular signaling pathways through protein-protein interactions to differential expression of proteins in response to external stimuli . We assess the relative merits of the different methods used and discuss issues and future directions in the field.

Wei Sheng Yan Jiu, 2002 Oct, 31(5), 398 - 401
{Recent advances for research on chemopreventive effect of beta-carotene}; Zhong W; Cancer is the culmination of the chronic disease process, and can be attributed the interactions of environment and genes . An optimistic message for 35% cancer prevention may be in dietary constituents, food supply . Carotenoids are isoprenoid compounds synthesized in plants and microorganisms, but not in animals . Human tissue contains only a fraction of the total number of carotenoids(nearly 600 have been identified in nature) present in food supply such as fruit and vegetables . People with high intake of beta-Carotene have a reduced risk of cancer . This outcome seems to bear relation to the chemical to quench singlet oxygen and inhibit peroxyl free radical reaction . But the ATBC and CARET cancer prevention study found the increase in lung cancer incidence in smokers and asbestos . This adverse effect may be explained by beta-Carotene prooxidant when tobacco smoke exposure and oxidative stress occurs, and cellular DNA damage was further aggrevated . Phase I enzymes were induced . Cell transformation was enhanced.

Aviakosm Ekolog Med, 2002, 36(5), 55 - 7
{Microbial destruction of harmful organic contaminants inherent to atmosphere of living space}; Nazarov NM et al.; The investigation into the microbial destruction of harmful organic contaminants in the air of habitable rooms was performed with the assumption that microorganisms have labile metabolism and synthesize inducible enzymes involved in transformation of organics compounds . Based on our observations, Methylobacterium D-08 and Arthrobacter pastens are able to synthesize inducible enzymes and transform ethanol, acetic acid, acetone, and aldehyde . Association of these microorganisms does not reveal acute antagonism in case of co-cultivation on mineral substrates with carbon provided by ethanol, acidic acid, acetone, and acetate aldehyde . The most rapid growth of microorganisms was observed at 28 degrees C and pH 7 . Determination of the effects of different concentrations of harmful organics on carbon dioxide production by the microbial association showed that the threshold concentration of acetate aldehyde was close to 2 mg/l and acetone--4 mg/l . The transformation activity was not reduced even at the ethanol and acidic acid concentrations of 80 and 100 mg/l, respectively . The association of A . pastens and M.D-08 oxidized ethanol, acidic acid, acetone and acetate aldehyde to carbon dioxide and water up to 90%.

J Hum Hypertens, 2003 Jan, 17(1), 21 - 7
Increased prevalence of Chlamydophila pneumoniae but not Epstein-Barr antibodies in essential hypertensives; Pitiriga VCh et al.; Conflicting data exist regarding the relationship between Chlamydophila pneunoniae (C . pneumoniae) and hypertension . In this study, both C . pneumoniae IgG and IgA titres and Epstein-Barr virus antibody levels were measured in 146 sustained hypertensives defined by 24 h ambulatory blood pressure monitoring (ABPM) and 54 normotensives . C . pneumoniae antibodies were measured by microimmunofluorescence test . IgG > or = 80 and IgA > or = 40 were defined as elevated antibody titres . Epstein-Barr antibodies were measured in order to investigate whether a possible association exists between hypertension and other, similarly widespread in the general population, intracellular microorganisms . All participants underwent casual blood pressure (BP) readings and 24 h ABPM . Subjects having mean 24 h systolic/diastolic ambulatory BP>125/80 mmHg, with or without antihypertensive medication were defined as hypertensives . Controls were free of any history or clinical evidence of hypertension, cardiovascular or pulmonary disease . Of the total participants, 77 hypertensives (52.7%) and 10 normotensives (18.5%) had IgA titres > or = 40 (crosstabs P < 0.000), whereas 76 hypertensives (52.1%) and 15 normotensives (27.8%) had IgG titres > or = 80, (crosstabs P < 0.002) . No difference was found in Epstein-Barr antibodies, between hypertensives and normotensives . In conclusion, C . pneumoniae, but not Epstein-Barr, antibody levels were found significantly higher in sustained hypertensives, suggesting high frequency of chronic C . pneumoniae, infections in this specific group of patients.

Appl Environ Microbiol, 2003 Feb, 69(2), 953 - 9
Growth of a Dehalococcoides-like microorganism on vinyl chloride and cis-dichloroethene as electron acceptors as determined by competitive PCR; Cupples AM et al.; A competitive PCR (cPCR) assay targeting 16S ribosomal DNA was developed to enumerate growth of a Dehalococcoides-like microorganism, bacterium VS, from a mixed culture catalyzing the reductive dehalogenation of cis-1,2-dichloroethene (cDCE) and vinyl chloride (VC), with hydrogen being used as an electron donor . The growth of bacterium VS was found to be coupled to the dehalogenation of VC and cDCE, suggesting unique metabolic capabilities . The average growth yield was (5.2 +/- 1.5) x 10(8) copies of the 16S rRNA gene/ micromol of Cl(-) (number of samples, 10), with VC being used as the electron acceptor and hydrogen as the electron donor . The maximum VC utilization rate (q) was determined to be 7.8 x 10(-10) micromol of Cl(-) (copy(-1) day(-1)), indicating a maximum growth rate of 0.4 day(-1) . These average growth yield and q values agree well with values found previously for dechlorinating cultures . Decay coefficients were determined with growth (0.05 day(-1)) and no-growth (0.09 day(-1)) conditions . An important limitation of this cPCR assay was its inability to discriminate between active and inactive cells . This is an essential consideration for kinetic studies.

Appl Environ Microbiol, 2003 Feb, 69(2), 835 - 44
Microbial community composition affects soil fungistasis; de Boer W et al.; Most soils inhibit fungal germination and growth to a certain extent, a phenomenon known as soil fungistasis . Previous observations have implicated microorganisms as the causal agents of fungistasis, with their action mediated either by available carbon limitation (nutrient deprivation hypothesis) or production of antifungal compounds (antibiosis hypothesis) . To obtain evidence for either of these hypotheses, we measured soil respiration and microbial numbers (as indicators of nutrient stress) and bacterial community composition (as an indicator of potential differences in the composition of antifungal components) during the development of fungistasis . This was done for two fungistatic dune soils in which fungistasis was initially fully or partly relieved by partial sterilization treatment or nutrient addition . Fungistasis development was measured as restriction of the ability of the fungi Chaetomium globosum, Fusarium culmorum, Fusarium oxysporum, and Trichoderma harzianum to colonize soils . Fungistasis did not always reappear after soil treatments despite intense competition for carbon, suggesting that microbial community composition is important in the development of fungistasis . Both microbial community analysis and in vitro antagonism tests indicated that the presence of pseudomonads might be essential for the development of fungistasis . Overall, the results lend support to the antibiosis hypothesis.

Curr Top Med Chem, 2003, 3(5), 471 - 84
Naturally occurring iminosugars and related compounds: structure, distribution, and biological activity; Asano N; Alkaloids mimicking sugars in size and shape are now believed to be widespread in plants and microorganisms . Iminosugars are monosaccharide analogs in which the ring oxygen has been replaced by an imino group . Such iminosugars inhibit the glycosidases involved in a wide range of important biological processes because of their structural resemblance to the sugar moiety of the natural substrate and the presence of the nitrogen atom mimicking the positive charge of the glycosyl cation intermediate in the enzyme-catalyzed glycoside hydrolysis . These iminosugars and their derivatives are arousing considerable attention as potential therapeutic agents . In this review, the distribution of naturally occurring iminosugars and their biological activities and therapeutic applications will be reviewed and the prospects of iminosugars and their derivatives for new therapeutic applications will also be described.

Cell Mol Life Sci, 2002 Dec, 59(12), 2097 - 111
Dictyostelium mobile elements: strategies to amplify in a compact genome; Winckler T et al.; Dictyostelium discoideum is a eukaryotic microorganism that is attractive for the study of fundamental biological phenomena such as cell-cell communication, formation of multicellularity, cell differentiation and morphogenesis . Large-scale sequencing of the D . discoideum genome has provided new insights into evolutionary strategies evolved by transposable elements (TEs) to settle in compact microbial genomes and to maintain active populations over evolutionary time . The high gene density (about 1 gene/2.6 kb) of the D . discoideum genome leaves limited space for selfish molecular invaders to move and amplify without causing deleterious mutations that eradicate their host . Targeting of transfer RNA (tRNA) gene loci appears to be a generally successful strategy for TEs residing in compact genomes to insert away from coding regions . In D . discoideum, tRNA gene-targeted retrotransposition has evolved independently at least three times by both non-long terminal repeat (LTR) retrotransposons and retrovirus-like LTR retrotransposons . Unlike the nonspecifically inserting D . discoideum TEs, which have a strong tendency to insert into preexisting TE copies and form large and complex clusters near the ends of chromosomes, the tRNA gene-targeted retrotransposons have managed to occupy 75% of the tRNA gene loci spread on chromosome 2 and represent 80% of the TEs recognized on the assembled central 6.5-Mb part of chromosome 2 . In this review we update the available information about D . discoideum TEs which emerges both from previous work and current large-scale genome sequencing, with special emphasis on the fact that tRNA genes are principal determinants of retrotransposon insertions into the D . discoideum genome.

Planta Med, 2003 Jan, 69(1), 9 - 14
Anti-inflammatory properties of piperlactam S: modulation of complement 5a-induced chemotaxis and inflammatory cytokines production in macrophages; Chiou WF et al.; Macrophages infiltrate tissues in response to chemoattractants including complement 5a (C5a) . Infiltrating macrophages clear microorganisms but also can cause tissue damage . We hypothesized that prevention of macrophages from excessive recruitment into infected sites may underlie the anti-inflammatory effects of piperlactam S, an alkaloid isolated from Piper kadsura (Choisy) Ohwi . To test this hypothesis, chemotactic migration of RAW264.7 macrophages was induced by C5a and the effects of piperlactam S were studied . The results showed that piperlactam S (1-30 microM) concentration-dependently suppressed C5a-induced migration across a fibrinogen-coated barrier with an IC50 of 4.5 +/- 0.3 microM . At 30 microM, piperlactam S inhibited chemotaxis by more than 95 % and also decreased phagocytosis by 25 % without reducing macrophage viability and adherent capacity . Furthermore, piperlactam S treated cells adhered but failed to spread and elongate as in control cells . Finally, piperlactam S inhibited the C5a-stimulated release of tumor necrosis factor-alpha and interleukin-1beta . We conclude that retardation of macrophage recruitment by interfering with the migration process and suppression of cytokines production might underlie the potential usefulness of piperlactam S as an anti-inflammatory agent.

Ophthalmologica, 2003 Jan-Feb, 217(1), 53 - 7
Elevated adrenomedullin in the vitreous of patients with diabetic retinopathy; Ito S et al.; Adrenomedullin (AM) is a multifunctional peptide with various physiological actions, including vasodilatation, a defense mechanism against microorganisms, the regulation of growth and the regulation of insulin and glucose . In this study, we measured the vitreous AM levels in patients with diabetes mellitus to determine its potential involvement in the pathogenesis of diabetic retinopathy (DR) . We used an immunoradiometric assay to measure the vitreous AM concentrations in a total of 28 eyes: 13 with DR and 15 with macular holes (15 men and 13 women, 62.9 +/- 10.4 years old) . The AM levels in the vitreous fluid of patients with DR (22.9 +/- 7.9 fmol/ml) were found to be significantly higher than the corresponding AM levels in patients with macular holes (4.7 +/- 1.1 fmol/ml) (p < 0.05) . These results indicate that the increase in the vitreous AM is related to DR .

Proc Natl Acad Sci U S A, 2003 Feb 18, 100(4), 1803 - 7 Epub 2003 Jan 31.
Facultative bacterial symbionts in aphids confer resistance to parasitic wasps; Oliver KM et al.; Symbiotic relationships between animals and microorganisms are common in nature, yet the factors controlling the abundance and distributions of symbionts are mostly unknown . Aphids have an obligate association with the bacterium Buchnera aphidicola (the primary symbiont) that has been shown to contribute directly to aphid fitness . In addition, aphids sometimes harbor other vertically transmitted bacteria (secondary symbionts), for which few benefits of infection have been previously documented . We carried out experiments to determine the consequences of these facultative symbioses in Acyrthosiphon pisum (the pea aphid) for vulnerability of the aphid host to a hymenopteran parasitoid, Aphidius ervi, a major natural enemy in field populations . Our results show that, in a controlled genetic background, infection confers resistance to parasitoid attack by causing high mortality of developing parasitoid larvae . Compared with uninfected controls, experimentally infected aphids were as likely to be attacked by ovipositing parasitoids but less likely to support parasitoid development . This strong interaction between a symbiotic bacterium and a host natural enemy provides a mechanism for the persistence and spread of symbiotic bacteria.

J Egypt Soc Parasitol, 1999, 29(3), 745 - 61
Mites infesting two migratory birds, Coturnix c . coturnix (quail or Simmaan) and Sturnus v . vulgaris (starling or zarzuur) with reference to avian zoonosis; Mazyad SA et al.; Birds are a marvelous group of creatures . Their beautiful coloration, singing, dancing and their attractive ways of life offer great pleasure to birdwatcher . Egypt is one of the most important countries of migratory birds . Not less than 300 species of birds visit Egypt annually from allover the world . The mite fauna of two migratory birds was studied in North Sinai Governorate and Suez Canal Zone . Thirty-one species of mites were recovered from quail and 39 from starling . Both types of birds serve hosts for 26 species of mites . Besides, five species were only recovered from quail and 13 species from starling . These totaled 44 species belonging to 30 families of three suborders (Mesostigmata, Trombidiformes and Sarcoptidiformes) . The mite index on quail ranged between 1.0 to 5.0 in North Sinai G . and 1.0 to 17.0 in Suez Canal Z . The mite index on starling ranged between 1.0 to 4.75 in North Sinai G . and 1.0 to 4.5 in Suez Canal Z . Sixteen of the recovered species of mites were not recorded before on the Egyptian resident birds (house sparrow and laughing dove) . The medical and veterinary importance of avian zoonosis was discussed . It is hoped to stimulate the awareness to migratory birds as reservoir hosts for microorganisms and parasites from allover the world . Besides, there is an urgent need to protect the resident and the visiting birds.

Ying Yong Sheng Tai Xue Bao, 2002 Sep, 13(9), 1179 - 86
{Advances in studies on the effect of surfactant on bioavailability of polycylcic aromatic hydrocarbons (PAHs) in soil}; Jiang X et al.; The solubility and adsorption/desorption equilibrium of PAHs in soil and their interaction with soil bacterium can be altered by surfactants, which lead to the alternation of PAHs bioavailability . The bioavailability of PAHs can be enhanced by the decrease of interface tension between soil and water, the increasement of PAHs' solubility, and the transportation facilitation of PAHs in the presence of surfactants . It also can be inhibited by the surfactant toxicity to the bacteria or by the competitive ultilization between non-toxicitic surfactants and PAHs by the bacteria . In addition, the effects of surfactants on the PAHs of different existence-forms in soils are different . The bioavailability of PAHs can be affected by the type and concentrion of surfactants, PAHs and soil microorganisms, and also by soil physi-chemical characteristics.

Ying Yong Sheng Tai Xue Bao, 2002 Sep, 13(9), 1169 - 73
{Effect of atmospheric CO2 and temperature increment on crop physiology and ecology}; Li F et al.; The effect of atmospheric CO2 and temperature increment on crop growth, photosynthesis, yield, and water and nutrient use efficiency was summarized and discussed in this paper . Under high CO2 concentration in the future, the photosynthetic rate of crops may be enhanced to some extent, and their biomass and yield may also be increased . Their stomatal conductance may be reduced, while water use efficiency (WUE) may be increased under elevated CO2 concentration . The aboveground biomass and root production, especially fine root production, will be greatly enhanced, and consequently, litter production increased, C/N ratios increased, plant litter decomposition rates declined under CO2 enrichment . Plants grown under elevated CO2 concentration may release more substrate into rhizosphere, which may lead to an increased activity of soil microorganisms, acclerated mineralization of nutrients, and improved nutritional status of the plant.

Environ Microbiol, 2003 Feb, 5(2), 133 - 8
Testing the limits of biological tolerance to arsenic in a fungus isolated from the River Tinto; Canovas D et al.; The Tinto river in Spain, with its high acidity and heavy metal concentrations (As, Cu, Cr, Zn), is an example of an environment hostile to life . Yet despite these extreme conditions, the site possesses a great diversity of eukaryotic life forms . We report the isolation of a filamentous fungus able to grow at 200 mM arsenic ( approximately 15 000 p.p.m.), i.e . a concentration 20-fold above that withstood by the reference microorganisms Escherichia coli, Saccharomyces cerevisiae and Aspergillus nidulans, and 200 times greater than that tolerated by Aspergillus niger . Based on morphological, physiological and genotypic criteria, the strain belongs to the genus Aspergillus . High concentrations of the metalloid induced vacuolation, suggesting that this organelle is someway connected to arsenic tolerance . Concentrations that are lethal to other organisms do not stress Aspergillus sp . P37 . The fungus was capable of removing arsenic from culture media . In addition to arsenic hyper-resistance, it also displayed a polyresistant phenotype to copper and chromium.

Res Microbiol, 2002 Dec, 153(10), 659 - 66
Expression of UreI is required for intragastric transit and colonization of gerbil gastric mucosa by Helicobacter pylori; Mollenhauer-Rektorschek M et al.; Helicobacter pylori colonizes the antral mucosa of the human stomach . There is a controversy as to whether the microorganism is exposed to acidity in its ecological niche . In vitro, the microorganism requires urease for gastric colonization and survival at pH < 4.0 . UreI encodes an acid activated urea channel enabling urea access to intrabacterial urease at acidic pH . UreI is also necessary for survival at pH < 4.0 . However, the role of UreI for both intragastric transit and colonization of the epithelial gastric mucosa has never been analyzed in detail . We therefore infected gerbils, whose intragastric pH and response to infection resemble those of man, with H . pylori G1.1 wild type bacteria and their corresponding isogenic ureI mutants . Inhibitors of gastric acid secretion and colonization were used for manipulation of gastric pH . Gastric colonization was determined by urease assay and PCR . Gastric pH was measured with pH electrodes . Whereas H . pylori wild type or ureI complemented ureI knockout bacteria colonized the antrum, ureI deletion mutants were unable to colonize . However, continuous inhibition of acid secretion resulted in gastric colonization by the ureI mutants, as also observed with the wild type strain . Restoration of acid secretion resulted in eradication of ureI mutants but not wild type bacteria . The data show that ureI is essential for both gastric transit after inoculation and mucosal colonization in the untreated stomach . The eradication of ureI mutants following restoration of acid secretion suggests that the organism is exposed to pH < 4.0 at the surface of the antral mucosa and that UreI provides a target for specific monotherapy of H . pylori infections.

Mikrobiol Z, 2002 Sep-Oct, 64(5), 35 - 46
{Effect of some electron acceptors on metabolism of methanogenic biotic community}; Chernyshenko DV; The dynamics of redox potential, pH, CH4, H2, CO2, N2 formation under the community development under anaerobic conditions in presence of acceptors of electrons (potassium ferricyanide, molecular oxygen, sodium nitrate and sodium sulphate) has been investigated . It has been established that the optimal value at the redox potential under the development of methanogenic community was created at the expense of microorganisms vital activity . The decrease of methanogenesis under the electron acceptors introduction is connected with the increase of the culture medium redox potential.

Can J Microbiol, 2002 Nov, 48(11), 955 - 64
Soil and plant effects on microbial community structure; Buyer JS et al.; We investigated the effects of two different plant species (corn and soybean) and three different soil types on microbial community structure in the rhizosphere . Our working hypothesis was that the rhizosphere effect would be strongest on fast-growing aerobic heterotrophs, while there would be little or no rhizosphere effect on oligotrophic and other slow-growing microorganisms . Culturable bacteria and fungi had larger population densities in the rhizosphere than in bulk soil . Communities were characterized by soil fatty acid analysis and by substrate utilization assays for bacteria and fungi . Fatty acid analysis revealed a very strong soil effect but little plant effect on the microbial community, indicating that the overall microbial community structure was not affected by the rhizosphere . There was a strong rhizosphere effect detected by the substrate utilization assay for fast-growing aerobic heterotrophic bacterial community structure, with soil controls and rhizosphere samples clearly distinguished from each other . There was a much weaker rhizosphere effect on fungal communities than on bacterial communities as measured by the substrate utilization assays . At this coarse level of community analysis, the rhizosphere microbial community was impacted most by soil effects, and the rhizosphere only affected a small portion of the total bacteria.

Indian Pediatr, 2003 Jan, 40(1), 36 - 40
A study of Vitamin K status in children on prolonged antibiotic therapy; Bhat RV et al.; Vitamin K deficiency is known to cause coagulopathy and bleeding in patients on prolonged antibiotic therapy . This study was conducted to evaluate the status of vitamin K deficiency in hospitalized children on prolonged antibiotic therapy and its role in reversing the coagulopathy . A prospective non-randomized study was conducted on children on antibiotic therapy at a tertiary care hospital . Children in the 1 month-1 year age group developed significant coagulopathy as compared to other age groups . Coagulation abnormalities were also seen to be more in children with greater grades of malnutrition, on a more prolonged course of antibiotics and in children who were critically ill in intensive care . Hypoprothrombinemia previously reported to be due to B-lactam antibiotics containing the N-Methyl Thio Tetrazole (NMTT) group also resulted from antibiotics without this side chain . Inhibition of intestinal microorganisms by antibiotics was thought to be a likely explanation of this phenomenon . We suggest Vitamin K prophylaxis in severely ill patients, on extended periods of antibiotics and inadequate diet to prevent morbidity and mortality.

J Invest Surg, 2003 Jan-Feb, 16(1), 23 - 8
Disruption of the small-intestine mucosal barrier after intestinal occlusion: a study with light and electron microscopy; Kabaroudis A et al.; It is known that the gut may serve as a reservoir for various microorganisms, which under specific circumstances may intrude into the systemic circulation, causing systemic infections . The aim of the present study was to estimate the "critical time" of disruption of the small-intestine mucosal barrier in conditions of experimentally induced intestinal occlusion, based on the histopathological alterations observed under light and electron microscopy . Forty rabbits underwent small-intestine obstruction through ligation with a nonabsorbable suture . Blood cultures from portal vein and inferior vena cava, as well as cultures from the peritoneal fluid, a hepatic fragment, and a mesenteric lymph node, were obtained before the ligation (0 h) . The same cultures were repeated at 4 and 8 h (group A, 20 rabbits) and at 6 and 12 h after the ligation (group B, 20 rabbits) . Small-intestine specimens proximal to the occlusion were taken for examination under the optic and electronic microscope in the same time intervals . Five of 20 rabbits of group A died within 4 h and 6 of 20 rabbits of group B died within 6 h after the operation . All anaerobic cultures were negative . All aerobic cultures that became positive developed Escherichia coli colonies . Intestinal epithelium of dead animals was transformed to cuboid with destruction of goblet cells and alteration in secretion of acid polysaccharides . The mucosal appearance of all rabbits that survived 12 hours after ligation was the same . The disruption of the mucosal barrier begins 4 h after complete intestinal occlusion . At 12 h after complete intestinal occlusion, the disruption is total with different degrees of severity.

Ugeskr Laeger, 2002 Dec 9, 164(50), 5911 - 5
{Autoimmune thyroiditis--an infectious disease?}; Rasmussen AK et al.; The aim was to review existing evidence of a possible role of infectious agents in the pathogenesis of autoimmune thyroid disease . Autoimmune thyroid disease is a polygenic, multifactorial disease in which genetically susceptible individuals are exposed to an environmental insult resulting in immune system activation . Different viruses (influenza B, rubella, retrovirus) have been associated with thyroiditis, but no single agent appears to be causative . There is no firm evidence of infection being an important trigger of autoimmune thyroid disease, and it has not been possible to isolate a microorganism neither by culture nor by molecular identification . Infection may be a precipitating factor in the development of autoimmune thyroid disease.

J Infect Dis, 2003 Feb 1, 187(3), 518 - 21 Epub 2003 Jan 24.
Detection of Ureaplasma urealyticum in second-trimester amniotic fluid by polymerase chain reaction correlates with subsequent preterm labor and delivery; Gerber S et al.; Ureaplasma urealyticum is the microorganism most frequently isolated from the amniotic fluid of women in preterm labor . The relationship between intra-amniotic U . urealyticum in healthy second-trimester pregnant women and subsequent pregnancy outcome was investigated . Transabdominal amniotic fluid obtained from 254 asymptomatic women at 15-17 weeks' gestation were tested by polymerase chain reaction (PCR) . U . urealyticum was identified in 29 subjects (11.4%) . A subsequent preterm labor occurred in 17 U . urealyticum-positive women (58.6%), compared with 10 (4.4%) U . urealyticum-negative women (P<.0001) . Preterm birth was documented in 7 (24.1%) U . urealyticum-positive women compared with only 1 U . urealyticum-negative woman (0.4%) (P<.0001) . U . urealyticum-positive women also had a higher prevalence of preterm labor in a prior pregnancy (20.7%) than did the negative women (2.7%; P=.0008) . PCR testing of second-trimester amniotic fluid for U . urealyticum can identify women at risk for subsequent preterm labor and delivery.

Wei Sheng Wu Xue Bao, 2001 Feb, 41(1), 49 - 53
{Determination of chromosome of Gluconobacter oxydans SCB329}; Lin W et al.; After the pure culture of Gluconobacter oxydans SCB329 was researched, its growth curve was measured and its logarithmic phase was determined as between 4-24 h . After the microorganisms were havested in its logarithmic phase, The intact chromosome was prepared by agarose-embedded method . Then the genome of SCB329 was analyzed by Pulsed-field Gel Electrophoresis . The result indicated that there are one chromosome and one great plasmid . The length of intact chromosome of SCB329 has been estimated to be approximately between 2.2 Mb and 3.5 Mb.

Shi Yan Sheng Wu Xue Bao, 2000 Sep, 33(3), 207 - 14
{Cloning and tissue expression pattern analysis of the human citrate synthase cDNA}; Liu Q et al.; Tricarboxylic acid (TCA) cycle is an important way to generate ATP, which is widely distributed in the cells of animal, plant or microorganism . It catalyses the catabolism of sugar as well as protein and fat . Citrate synthase plays a key role in regulating TCA cycle and is responsible for catalysing the synthesis of citrate from oxaloacetate and acetyl CoA . Screening of genomic informatics was performed by using pig citrate synthase cDNA as a probe and a contig which is 1636 bp long and has highly homologous to the pig citrate synthase cDNA was obtained from selected ESTs with the ASSEMBLY program . According to the sequence of this contig, a pair of primers was designed and used to amplify cDNA libraries . A 1492 bp cDNA containing an open reading frame encoding 466 amino acids was cloned from human testis and skeletal muscle cDNA libraries . The deduced amino acid sequence of the cDNA showed 95%, 92% and 60.9% identity to pig, chicken and yeast citrate synthase respectively . Because the deduced amino acids sequence contains a highly conserved motif of citrate synthase from three different species, it is believed that this cDNA may be a transcript of human citrate synthase gene . Northern analysis showed that the human citrate synthase was expressed at high level in heart and muscle, at middle level in brain, kidney and pancreas tissues, not detectable in thymus and small intestine tissues, and at low level in other nine tested human tissues.

Wei Sheng Wu Xue Bao, 2001 Apr, 41(2), 191 - 7
{Biosynthesis of glutathione: construction of ATP regeneration system between recombinant E . coli and S . cerevisiae}; Li Y et al.; An ATP regeneration system between two microorganisms, in which recombinant E . coli II-1 and S . cerevisiae WSH-J702 were used as the biosynthesis enzymes source of glutathione and ATP, respectively, was constructed by using glucose as an energy substrate . Yeast cells after permeable treatment could hardly consume glucose . The glycolysis pathway of yeast was started by adding 1.0 mmol/L AMP and 0.05 mmol/L NADH simultaneously . GSH biosynthesis in the coupling system was improved with increasing glucose concentration to 400 mmol/L, where the concentration of GSH reached 10.4 mmol/L (3.2 g/L) . Both the coupling system and the non-coupling system with ATP added could hardly produce GSH under the absence of magnesium . The low efficiency of ATP regeneration system constructed was probably caused by the chelation between magnesium and ATP . By adding magnesium to the coupling system, a final concentration of GSH, 14.3 mmol/L (4.4 g/L), was achieved at 6 h of reaction.

Wei Sheng Wu Xue Bao, 2001 Apr, 41(2), 133 - 40
{Extraction of DNA from environmental samples and construction of mixed genomic DNA libraries}; Wang X et al.; A method has been developed for extracting and purifying genomic DNA from environmental samples . In this method, an environmental sample is treated first by grinding and freezing/thawing and subsequently by SDS/proteinase K-based DNA extraction . The yields of purified DNA from three samples used in this study ranged from 2 to 16 micrograms per gram of dry sample . Mixed genomic DNA libraries for two of the environmental samples were constructed by inserting restriction fragments (3-8 kb) of the purified DNAs into plasmid pUC18 and transforming E . coli DH5 alpha with the resultant plasmids . Approximately 10(3) to 10(4) insert-containing clones were obtained from 1 g of each sample . Clone libraries were analyzed by DNA sequencing and gene annotation . Among 20 randomly-selected clones, 14 contained an insert whose sequence had not been reported while the rest had an insert of either E . coli or vector origin . A search of sequence databases using the end sequences of each of the foreign inserts showed that each sequence was part of a gene encoding, in most cases, a predictable function . Our results are of significance to the collection, investigation and exploitation of the genes of uncultured microorganisms.

Wei Sheng Wu Xue Bao, 1998 Dec, 38(6), 411 - 6
{Microbial succession on lignite along with weathering}; Yuan H et al.; Six different weathered lignite samples were examined by scanning electron microscopy . Few microorganisms were observed on lignit just excavated and only spores and short hyphae were observed on lignite samples excavated 5 months, 1 year and 4 years before sampling . When lignite samples were moistened with distilled water and incubated for 10 days, actinomycetes proliferated significantly on lignite samples that were just excavated or excavated 5 months before sampling . The growth of bacteria was observed on lignite samples excavated 1 year before sampling . Fungi increased in length and in number on lignite samples excavated 4 years before sampling . These findings of microbial succession on lignite samples along with weathering were consistent with results of the plate count method; actinomycetes are the first colonizers, then bacteria and fungi are the last degrader . The dominant microorganisms were Actinosynnema sp., Streptomyces sp . and Nocardia sp . among actinomycetes, two Arthrobacter sp . among bacteria and two Aspergillus sp . among fungi.

Br J Sports Med, 2003 Feb, 37(1), 76 - 9
Effects of 12 months of exercise training on salivary secretory IgA levels in elderly subjects; Akimoto T et al.; BACKGROUND: The immune system declines in efficiency with advancing age, making the elderly less resistant to pathogenic microorganisms . Upper respiratory tract infection (URTI) is a common illness . Recent studies have shown that suppression of secretory immunoglobulin A (SIgA) is associated with increased incidence of URTI . OBJECTIVE: To assess the effect of exercise on salivary SIgA in elderly subjects . METHODS: Forty five elderly subjects (18 men, 27 women; mean (SD) age 64.9 (8.4) years) performed both 60 minute resistance and 60 minute moderate endurance training a week for 12 months . Saliva samples were obtained before training, and at four and 12 months during the training period . Salivary SIgA concentrations were measured by enzyme linked immunosorbent assay, and the SIgA secretion rate was calculated . RESULTS: SIgA concentrations before training, and at four and 12 months during training were 24.7 (14.4), 27.2 (14.2), and 33.8 (18.5) micro g/ml respectively . SIgA secretion rates were 29.5 (26.0), 33.8 (27.2) and 46.5 (35.1) micro g/min respectively . The results indicate that both the concentration and secretion rate of SIgA significantly (p<0.01) increased during 12 months of exercise in these elderly subjects . CONCLUSION: Regular moderate exercise seems to enhance mucosal immune function in elderly subjects.

Math Biosci, 2003 Mar, 182(1), 81 - 92
Design principles and operating principles: the yin and yang of optimal functioning; Voit EO; Metabolic engineering has as a goal the improvement of yield of desired products from microorganisms and cell lines . This goal has traditionally been approached with experimental biotechnological methods, but it is becoming increasingly popular to precede the experimental phase by a mathematical modeling step that allows objective pre-screening of possible improvement strategies . The models are either linear and represent the stoichiometry and flux distribution in pathways or they are non-linear and account for the full kinetic behavior of the pathway, which is often significantly effected by regulatory signals . Linear flux analysis is simpler and requires less input information than a full kinetic analysis, and the question arises whether the consideration of non-linearities is really necessary for devising optimal strategies for yield improvements . The article analyzes this question with a generic, representative pathway . It shows that flux split ratios, which are the key criterion for linear flux analysis, are essentially sufficient for unregulated, but not for regulated branch points . The interrelationships between regulatory design on one hand and optimal patterns of operation on the other suggest the investigation of operating principles that complement design principles, like a user's manual complements the hardwiring of electronic equipment.

Sheng Li Ke Xue Jin Zhan, 2001 Jul, 32(3), 233 - 9
{The role of immune cells and inflammatory mediators in pathogenesis of intestinal inflammation}; Ding GF; The initiators of intestinal inflammation are greatly variable, but the mechanisms underlying the immunologically mediated mucosal damage are similar . A great progress in our understanding of the functions of gut-associated lymphoid tissue was achieved because of the advances in immuno-biology and molecular immunology . At the beginning of an inflammation microorganisms and hither to derived products or allergen firstly stimulate and activate the specific and nonspecific immune cells, and then intestine epithelials, macrophages and lymphocytes release various cytokines . These cytokines are able to recruit more immune cells to be activated and more cytokines to be released . A "cascade" is then generated between epithelials, macrophages and lymphocytes . Finally the inflammation in certain area of intestinal tract is generated . The balance between inflammatory factors and anti-inflammatory molecules depends the results of the inflammation . Therefore, the better understanding of the mechanisms on inflammatory and anti-inflammatory factors is helpful for the diagnosis and treatment of intestinal inflammation.

Wei Sheng Yan Jiu, 2002 Jun, 31(3), 160 - 2
{Variations and transformations of nitrogen pollutants in source and tap water of D-lake}; Liu D et al.; Ammonia, nitrite and nitrate pollutions and variations in source water and tap water of D-Lake are investigated . The removal effects of nitrogen in routine proceeding of water production and the transformations of the different forms of nitrogen in natural condition with simulated test are studied at the same time . The results show that the nitrogen pollutions in source and tap water of D-waterworks are higher than T-waterworks', although both waterworks have same resource . Compared with D-waterworks, the nitrogen pollutions of source and tap water in Z-waterworks are lower than D-waterworks' and they have different variation and distribution . The forms of nitrogen could be partly transformed during the routine production, but the total percentage of nitrogen removal efficiency is not significant (just about 20%) . The simulate test shows that NH4+(-)N could change into NO2-(-)N, while No2-(-)N into NO3-(-)N, which is partly transformed into nitrogen gas by microorganism and majority of which are remained in tap water by form of NO3-(-)N.

Am J Epidemiol, 2003 Feb 1, 157(3), 227 - 33
Exposure to metal fume and infectious pneumonia; Palmer KT et al.; To test the hypothesis that inhalation of metal fume reversibly increases susceptibility to pneumonia, the authors conducted a case-control study . Men aged 20-64 years, admitted to 11 hospitals in West Midlands, England, with community-acquired pneumonia during 1996-1999 were interviewed about their lifetime occupational history, exposure to metal fume, and potential confounding factors . Similar information was collected from controls admitted to the same hospitals with nonrespiratory illness . For cases, exposures were timed relative to the onset of their illness (on average, 6 months before interview) . Exposure histories for controls were censored 6 months before interview . Interviews were completed by 525 cases and 1,122 controls (response rates of 74% and 99%) . Pneumonia was associated with reported occupational exposure to metal fume in the previous year (adjusted odds ratio (OR) = 1.6, 95% confidence interval (CI): 1.1, 2.4) but not in earlier periods (OR = 1.1) . The risk was highest for lobar pneumonia and recent exposure to ferrous fume (OR = 2.3, 95% CI: 1.2, 4.3) . The association was not specific to any one microorganism . These findings support the hypothesis that ferrous and possibly other metal fumes reversibly predispose to infectious pneumonia . Research should now focus on the underlying mechanisms and prevention.

Environ Microbiol, 2003 Jan, 5(1), 72 - 4
Monitoring the colonization of monuments by bacteria: cultivation versus molecular methods; Laiz L et al.; Building materials commonly used in wall paintings and monuments (mortar, limestone and sandstone) were inoculated with an artificial consortium composed of 14 microorganisms and incubated for 6 months at 28 degrees C . The colonization of the different materials by the consortium was investigated . Culture-independent techniques revealed the presence of a diversity of bacteria, whereas culture-dependent techniques yielded mainly spore-forming bacteria . The data suggest that plating leads to an overestimation of the number of spore-forming bacteria with respect to quiescent vegetative forms; the latter are less easily cultured, but are readily detected by culture-independent techniques.

Cell Microbiol, 2003 Jan, 5(1), 3 - 14
Insect immunity and its implication in mosquito-malaria interactions; Dimopoulos G; Insects' resistance to infectious agents is essential for their own survival and also for the health of the plant, animal and human populations with which they closely interact . Several of the major human diseases are spread by insects and are rapidly expanding as a result of the development of insecticide resistance in vectors and drug resistance in parasites . A vector insects' permissiveness to a pathogen, and hence the spread of the disease, will largely depend on the compatibility of the molecular interactions between the two species and the capability of the insect immune system to recognize and kill the pathogen . The innate immune system comprises a variety of components and mechanisms that can discriminate between different microorganisms and mount specific responses to control pathogenic infections . An impressive body of knowledge on the insects' innate immunity has been generated from studies in the model organism Drosophila . These studies are now guiding the exploration of the immune system in the vector mosquito of human malaria, Anopheles, and its implication in the elimination of parasites . Anopheles immune responses have been linked to parasite losses and some refractory mosquitoes can kill all parasites through specific defence mechanisms . The recently sequenced Drosophila and Anopheles genomes provide a detailed and comparative view on their immune gene repertoires that in combination with post-genomic analyses is used to further dissect the complex mechanisms of Plasmodium killing in the mosquito.

Appl Microbiol Biotechnol, 2003 Jan, 60(5), 556 - 9 Epub 2002 Dec 04.
Development of a low-cost technology for mass production of the free-living nematode Panagrellus redivivus as an alternative live food for first feeding fish larvae; Ricci M et al.; The free-living nematode Panagrellus redivivus is a suitable food source for first feeding fish . In the present report, a new method for the mass production of P . redivivus is presented . The technique involves multiplication of the nematode in monoxenic (single microorganism: Saccharomyces cerevisiae) solid culture (fluid media supported by 1- to 4-cm(3) sponge cubes) in autoclavable plastic bags (size range: 50 x 30 cm to 75 x 67 cm) . Two growing media were tested: oat-meal medium (OM), which is an oat-based medium (16.7% oat-meal flour in 0.8% saline solution), and purified ingredient medium (PIM), a semi-synthetic medium (1.64% meat peptone, 0.94% yeast extract, 12.6% corn starch, 0.24% glucose, 1.48% sunflower oil, in 0.8% saline solution) . The bags were inoculated with 350 nematodes/g medium . After an average period of 12 days (11-13 days) at 25 degrees C, the average yield (number of nematodes/g medium) was 241 x 10(3) for OM and 333 x 10(3) for PIM in 12-l bags (50 x 30 cm) . The production scale has currently reached a bag volume of 50 l (75 x 67 cm); using PIM and the conditions described above, it was possible to harvest more than 1.3 x 10(9) nematodes/bag (291 x 10(3) nematodes/g medium) . In PIM, when sun flower oil was replaced with the same amount of fish oil or cod liver oil, yields of 259 x 10(3) and 290 x 10(3) nematodes/g medium, respectively, were attained . The technology for mass production and formulation of P . redivivus should enable fish-hatchery operators to rely on a cheap, standardised, and permanently available live food product for first feeding fish larvae.

Immunol Cell Biol, 2003 Feb, 81(1), 52 - 8
Immune responses to hepatitis B surface antigen following epidermal powder immunization; Osorio JE et al.; Langerhans cells in the epidermis of skin are potent antigen-presenting cells that trigger the immune system to respond to invading microorganisms . We have previously shown that epidermal powder immunization with a powdered inactivated influenza virus vaccine, by targeting the Langerhans cell-rich epidermis, was more efficacious than deeper tissue injection using a needle and syringe . We now report enhanced humoral and cellular immune responses to recombinant hepatitis B surface antigen following epidermal powder immunization . We observed that epidermal powder immunization with unadjuvanted hepatitis B surface antigen elicited an antibody titre equivalent to that induced by the alum-adjuvanted vaccine delivered by intramuscular injection, suggesting that epidermal powder immunization can overcome the need for adjuvantation . We demonstrated that synthetic CpG oligonucleotides (CpG DNA) could be coformulated with hepatitis B surface antigen and delivered by epidermal powder immunization to further augment the antibody response and modulate T helper cell activities . Epidermal powder immunization of hepatitis B surface antigen formulated with CpG DNA formulations resulted in 1.5-2.0 logs higher IgG antibody titres than alum-adjuvanted commercial vaccines administered by intramuscular injection . Formulation of hepatitis B surface antigen with CpG DNA elicited an augmented IgG2a antibody response and increased frequency of IFN-gamma secreting cells . In addition, CpG DNA was found to activate epidermal Langerhans cells and stimulate the production of TNF-alpha and IL-12 cytokines by epidermal cells, explaining its strong adjuvant activity following epidermal powder immunization . These results show that epidermal powder immunization is a safe and effective method to deliver hepatitis B surface antigen and the addition of new adjuvants, such as CpG DNA, may further enhance the efficacy of this vaccine.

In Vitro Cell Dev Biol Anim, 2002 Jul-Aug, 38(7), 423 - 7
In vitro cultivation of Wolbachia in insect and mammalian cell lines; Noda H et al.; Wolbachia infecting the small brown planthopper, Laodelphax striatellus, were successfully maintained and cultivated in two insect and one mammalian cell lines . The bacteria with the planthopper ovary were introduced into the flasks with the cultures of the cell lines . The Wolbachia proliferated in mosquito (Aedes albopictus) and lepidopteran (Heliothis zea) cell lines and in the mouse cell line, L929 . Proliferation of Wolbachia was confirmed by electron microscopy and quantitative polymerase chain reaction . This simple method for the cultivation of Wolbachia was applicable to other strains of Wolbachia, such as the one found in the lepidopteran eggs, and should facilitate fundamental and applied studies of this important group of microorganisms.

Exp Cell Res, 2003 Jan 15, 282(2), 70 - 7
NADPH-oxidase activation in murine neutrophils via formyl peptide receptors; Bylund J et al.; Neutrophils play a key role at inflammatory sites where, in addition to destroying infecting microorganisms, they may also have deleterious effects on host tissues . Both activities involve activation of the NADPH-oxidase that produces bactericidal and tissue-destructive reactive oxygen species (ROS) . We activated the murine NADPH-oxidase using different types of neutrophil activators and characterized the oxidative responses with respect to magnitude, localization, and kinetics . We show that agonist-induced activation of murine neutrophils results exclusively in extracellular release of ROS and no intracellular production could be detected . We also show that the formylated peptide, formyl-Met-Leu-Phe (fMLF), is a much less potent activator of the murine NADPH-oxidase than of the human analogue . Nevertheless, fMLF responses can be primed by pretreating the murine neutrophils with either cytochalasin B or bacterial lipopolysaccharide . Finally, we show that a synthetic hexapeptide, WKYMVM, is a more potent stimulus than fMLF for murine neutrophils and that these two agonists probably act via nonidentical high-affinity receptors.

J Microbiol Methods, 2003 Mar, 52(3), 389 - 93
Removal of PCR inhibitors from soil DNA by chemical flocculation; Braid MD et al.; Extracting high-purity DNA directly from soil has become essential for the study of microorganisms in environmental samples . However, many soils contain compounds that inhibit enzymes involved in manipulating DNA . In this study, chemical flocculation using multivalent cations was investigated as a potential method for eliminating soil-based inhibitors during the extraction process . The addition of AlNH(4)(SO(4))(2) during extraction significantly reduced the co-purification of PCR inhibitors with minimal loss of DNA yield.

Vaccine, 2003 Jan 30, 21(7-8), 809 - 11
Targeting of plant-derived vaccine antigens to immunoresponsive mucosal sites; Rigano MM et al.; Most pathogenic microorganisms enter their host via the mucosal surfaces lining the digestive, respiratory and urino-reproductive tracts of the body . The most efficient means of protecting these surfaces is through mucosal immunization . Transgenic plants are safe and inexpensive vehicles to produce and mucosally deliver protective antigens . However, the application of this technology is limited by the poor response of the immune system to non-particulate, subunit vaccines . Co-delivery of therapeutic proteins with targeting proteins, such as the B subunit of the Escherichia coli heat labile enterotoxin (LTB), could increase the effectiveness of such antigens.

Water Res, 2003 Feb, 37(4), 753 - 62
Virus removal during simulated soil-aquifer treatment; Quanrud DM et al.; Removals of indigenous coliphage and seeded poliovirus type 1 during simulated soil-aquifer treatment were evaluated during transport of secondary effluent under unsaturated flow conditions in 1-m soil columns . Independent variables included soil type (river sand or sandy loam) and infiltration rate . Removal of coliphage was in all cases less than removal of poliovirus type 1 (strain LSc-2ab), supporting contentions that indigenous coliphage can act as a conservative indicator of groundwater contamination by viral pathogens of human origin . Coliphage retention was significantly more efficient (p<0.001) in the finer-grained sandy loam (93%) than in sand (76%) . Increasing reactor detention time from 5 to 20 h increased coliphage attenuation from 70% to 99% in a 1-m sand column . There was a significant linear correlation (p=0.012) between log-transformed (fractional) coliphage concentration {log(C/C(0))} and reactor detention time . Re-mobilization of attached coliphage occurred during simulated rainfall using low-ionic-strength water . Inhibition of aerobic respiration resulted in significantly less efficient coliphage attenuation (p=0.033), suggesting the involvement of aerobic microorganisms in the survival/retention of this virus.

Anal Biochem, 2003 Jan 15, 312(2), 182 - 90
Enhanced nucleic acid capture and flow cytometry detection with peptide nucleic acid probes and tunable-surface microparticles; Chandler DP et al.; New methods for automated, direct nucleic acid purification and detection are required for the next generation of unattended environmental monitoring devices . In this study we investigated whether tunable-surface bead chemistry and peptide nucleic acids (PNA) could enhance the recovery and detection of intact rRNA in both test tube and automated suspension array hybridization formats . Intact rRNA was easily captured and detected on PNA-coated Lumavidin beads from 0.1 ng total RNA with a 15-min hybridization in pH 7 buffer, representing 1.7 x 10(3) cell equivalents of total RNA . DNA-conjugated beads in pH 5 hybridization buffer required an overnight hybridization to achieve a detectable signal at 0.1 ng target RNA . Standard DNA hybridization conditions (pH 7) were one order of magnitude less sensitive than the tunable-surface (pH 5) condition . The PNA-conjugated particles were 100x more sensitive than the tunable-surface DNA particles in the automated format, with a detection limit of 0.1 ng total RNA . The detection limits for total RNA on PNA-conjugated microparticles is immediately conducive to the detection and characterization of microorganisms in low-biomass environments or to the identification of rare sequences in a complex sample mixture, without using PCR.

Astrobiology, 2002 Fall, 2(3), 281 - 92
Extremophiles and the search for extraterrestrial life; Cavicchioli R; Extremophiles thrive in ice, boiling water, acid, the water core of nuclear reactors, salt crystals, and toxic waste and in a range of other extreme habitats that were previously thought to be inhospitable for life . Extremophiles include representatives of all three domains (Bacteria, Archaea, and Eucarya); however, the majority are microorganisms, and a high proportion of these are Archaea . Knowledge of extremophile habitats is expanding the number and types of extraterrestrial locations that may be targeted for exploration . In addition, contemporary biological studies are being fueled by the increasing availability of genome sequences and associated functional studies of extremophiles . This is leading to the identification of new biomarkers, an accurate assessment of cellular evolution, insight into the ability of microorganisms to survive in meteorites and during periods of global extinction, and knowledge of how to process and examine environmental samples to detect viable life forms . This paper evaluates extremophiles and extreme environments in the context of astrobiology and the search for extraterrestrial life.

Vestn Khir Im I I Grek, 2002, 161(3), 11 - 5
{Electrostimulation therapy--a new method of preventing postoperative wound complications in abdominal surgery}; Lokhvitskii SV et al.; Experimental investigations have shown that chemotaxic activity of microorganisms in the parietal peritoneum and abdominal organ tissues is inhibited when they are treated with impulse electric discharges preventing the process of infecting the tissues . Results of the experiments underlay the development of a new method of prophylactics of postoperative complications in abdominal surgery based on the electroimpulsive treatment of the operation wound . Clinical approbation of the method showed its effectiveness, simplicity and safety.

Expert Rev Mol Diagn, 2003 Jan, 3(1), 105 - 15
Detection of Shiga toxin-producing Escherichia coli in food; Alexandre M et al.; Shiga toxin-producing Escherichia coli are emerging as a significant source of food-borne infectious disease all over the world . Illness caused by Shiga toxin-producing E . coli can range from self limited, watery diarrhea to life-threatening manifestations such as hemorrhagic colitis, hemolytic uremic syndrome or thrombotic thrombocytopenic purpura and death . Shiga toxin-producing E . coli can potentially enter the human food chain from a number of animal sources, most commonly by contamination of meat with feces or intestinal contents after slaughter or cross-contamination of unpasteurized milk products . Because of the low infectious dose of the O157:H7 Shiga toxin-producing E . coli strain, laboratory diagnosis of Shiga toxin-producing E . coli in food samples has developed a great importance . This review will focus on the microorganism, giving priority to illness prevention and Shiga toxin-producing E . coli detection in food.

Biochim Biophys Acta, 2003 Jan 3, 1625(1), 1 - 10
Genomic and molecular characterization of CL-43 and its proximal promoter; Hansen S et al.; Collectins are part of the innate immune system as they bind nonself glycoconjugates on the surface of microorganisms and inhibit infection by direct neutralization, agglutination or opsonization of the invaders . Conglutinin and CL-43 are serum proteins that have only been found and characterized in Bovidae . We have studied molecular and genomic characteristics of CL-43 to identify polymorphisms that might be associated with disease-susceptible phenotypes or other traits in cattle, and to elucidate how the Bovidae may benefit from possessing additional collectins . Screening a bovine cDNA library resulted in the isolation of two plasmid clones that encoded the entire translated sequence of CL-43 . The 5'-untranslated end and start point of transcription were identified by 5'-RACE and showed that the mRNA transcript comprises either 1326 or 1241 nucleotides because of alternative splicing . Both transcripts encode a protein of 321 amino acids including a signal peptide of 20 residues . Characterization of two overlapping genomic lambda phage clones showed that the gene comprised seven exons spanning 8.5 kbp . The CL-43 gene, like the conglutinin gene, was mapped to Bos taurus chromosome 28 at q1.8 . The CL-43 promoter has 96% identity with the conglutinin promoter recently described by us, and the assignment of potential cis-regulatory elements shows that several hepatic transcription factors may regulate transcription in the acute phase response and in response to metabolic changes.

Trends Microbiol, 2003 Jan, 11(1), 44 - 51
The cellular paradigm of chlamydial pathogenesis; Stephens RS; Diseases caused by Chlamydia are based on intense and chronic inflammation elicited and maintained by reinfection or persistent infection . The traditional view in the field is that disease is mediated by antigen-dependent delayed-type hypersensitivity or autoimmunity . This immunological paradigm has served as the basis for years of chlamydial research but the mechanism or the antigen that causes pathology has yet to be unequivocally revealed . Recent research on responses elicited in Chlamydia-infected cells defines a new direction for our understanding of this microorganism-host interaction and provides the basis for a reassessment of disease mechanisms . Chlamydia-infected non-immune mammalian cells produce proinflammatory chemokines, cytokines, growth factors and other cellular modulators . This cellular response to infection supports an alternative hypothesis for chlamydial pathogenesis: the inflammatory processes of chlamydial pathogenesis are elicited by infected host cells and are necessary and sufficient to account for chronic and intense inflammation and the promotion of cellular proliferation, tissue remodeling and scarring, the ultimate cause of disease sequelae.

J Control Release, 2003 Jan 17, 86(2-3), 339 - 48
Gene expression and mucosal immune responses after vaginal DNA immunization in mice using a controlled delivery matrix; Shen H et al.; IgA antibodies in the vaginal tract are essential as a first defense line against microorganisms that enter the body via mucosal surfaces . Several studies have shown that direct application of DNA to the vaginal mucosal surface can induce secretion of IgA molecules specific to the expressed protein . The major challenge of formulating effective vaccines is to overcome the barriers to DNA administration caused by the estrus cycle and physical environment of the vaginal tract . In this study, we investigated whether controlled delivery of DNA to the vaginal surface would induce long-term IgA antibody production by applying controlled delivery matrices to the vaginal tract . The controlled DNA delivery matrices were composed of poly(ethylene-co-vinyl acetate) (EVAc) and loaded with a model plasmid encoding sperm-specific lactate dehydrogenase C(4) (LDH-C(4)) . These EVAc matrices provided a controlled and sustained DNA release to the vaginal mucosal surface . The DNA released from the EVAc disks was functionally active and capable of transfecting vaginal tissues . When inserted into the vaginal tract of mice, the DNA-loaded EVAc matrices triggered the immune system and induced specific IgA to LDH-C(4) in the vaginal secretions . These results demonstrate that the EVAc disks are efficient and convenient vehicles for delivering DNA to the vaginal tract and providing long-term local immunity.

Mikrobiologiia, 2002 Nov-Dec, 71(6), 836 - 48
{Detection of microorganisms in the environment and the preliminary appraisal of their physiological state by X-ray microanalysis}; Muliukin AL et al.; The paper deals with the X-ray microanalysis of the elemental composition of bacteriomorphic particles in 170,000-year old Antarctic permafrost sediments and in indoor dust . A comparative analysis of the phosphorus, sulfur, calcium, and potassium contents and the Ca/K and P/S ratios in these particles and in reference microbial cells occurring in different physiological states showed that the absence of P and/or S peaks in the X-ray spectrum of an object may indicate that it is abiotic . Resting microbial forms can be revealed on the basis of the following characteristic features: an increased content of Ca, a high Ca/K ratio, and a low P/S ratio . Model experiments with nonviable bacterial and yeast micromummies with alterations in the structural and barrier functions of the cytoplasmic membrane showed that micromummies can be recognized by a super-high content of a marker element (e.g., P, K, or Si), accumulated due to facilitated diffusion along the deliberately created concentration gradient . Such an analysis of the permafrost sediment and dust made it possible to suggest the presence of mummified cells in these objects . The possibility of using X-ray microanalysis for the detection of microbial cells in natural habitats in order to enhance the efficiency of ecological monitoring of the environment is discussed.

Eur J Clin Microbiol Infect Dis, 2002 Dec, 21(12), 890 - 1 Epub 2002 Dec 11.
Eikenella corrodens infective endocarditis in a previously healthy non-drug user; Watkin RW et al.; Reported here is a case of Eikenella corrodens aortic valve infective endocarditis presenting as a stroke in a previously healthy 31-year-old man . The patient had no evidence of structural heart disease and reported no history of intravenous drug use or recent dental treatment . There are no other cases reported in the available literature in which this microorganism has caused endocarditis in the absence of recognised risk factors.

J Virol, 2003 Feb, 77(3), 1992 - 2002
Molecular and functional analysis of an interferon gene from the zebrafish, Danio rerio; Altmann SM et al.; The interferon (IFN) family consisting of alpha IFN (IFN-alpha), IFN-beta, IFN-omega, IFN-delta, IFN-kappa, and IFN-tau is a large group of cytokines involved in the innate immune response against various microorganisms . Genes for IFN have been cloned from a variety of mammalian and avian species; however, IFN genes from lower-order vertebrates have not been forthcoming . Here, we report the cloning and characterization of the IFN gene from the zebrafish, Danio rerio . Zebrafish IFN (zfIFN) is 185 amino acids in length, with the first 22 amino acids representing a putative signal peptide . Treatment with the known IFN inducer polyinosinic acid-polycytidylic acid (poly{I}-poly{C}) resulted in an increase in zfIFN mRNA transcripts . zfIFN was also able to activate the IFN-inducible Mx promoter when cotransfected with a plasmid containing the zebrafish Mx promoter upstream of a luciferase reporter gene . To demonstrate antiviral activity, zebrafish cells were transfected with zfIFN and challenged with a fish rhabdovirus . A 36% reduction in plaque number was seen in zfIFN-transfected cells, compared to cells transfected with a control vector . Phylogenetic analysis has shown zfIFN to be approximately equally divergent from avian and mammalian IFN, consistent with its origin from an IFN present in the most recent common ancestor of these divergent lineages . A putative IFN from puffer, Fugu rubripes, was also found when zfIFN was used to search the fugu genome database, demonstrating that zfIFN can be used to find additional fish IFN genes . These results demonstrate that zebrafish can be used as an effective model for studying innate immunity and immune response to infectious disease.

Semin Neurol, 2002 Jun, 22(2), 133 - 42
Infectious myelopathies; Berger JR et al.; Although infectious myelopathies are rare, timely and accurate diagnosis is essential to improving outcome . There are a number of organisms that may cause infectious myelopathies, including human immunodeficiency virus (HIV), human T-cell lymphotropic virus type I (HTLV-I), herpesviruses, enteroviruses, Treponema pallidum, Mycobacterium tuberculosis, fungi, and parasites . Vacuolar myelopathy, the most common form of spinal cord disease in HIV-infected individuals, is underrecognized clinically . The failure to diagnose this condition is generally a consequence of the attribution of the lower extremity weakness and paresthesias to general debility and concomitant peripheral neuropathy . Tropical spastic paraparesis or HTLV-I-associated myelopathy involves the pyramidal tracts, chiefly at the thoracic level, and results in spastic lower extremity weakness and a spastic bladder . The herpesviruses (varicella-zoster, herpes simplex type 2, cytomegalovirus) and the enteroviruses cause myelitis . Prior to the development of antibiotics, syphilis was the most frequent infectious cause of spinal cord disease . In light of the broad spectrum of pathogens that may affect the spinal cord and the variegate fashion in which these disorders may present, the physician must always consider an infectious etiology in the differential diagnosis for the patient presenting with myelopathy . This review addresses the infectious myelopathies by microorganism.

Nat Struct Biol, 2003 Feb, 10(2), 126 - 30
Nickel coordination is regulated by the DNA-bound state of NikR; Carrington PE et al.; The uptake of nickel in Escherichia coli and other microorganisms is transcriptionally regulated by the NikR repressor or its homologs . Here we report the structure of the high-affinity nickel-binding site in NikR and show that it responds dramatically to DNA binding . X-ray absorption spectroscopy reveals that nickel in the holo-NikR protein is bound in a novel four-coordinate planar site consisting of two histidines, one additional O- or N-donor ligand and one S-donor ligand . Site-directed mutation of His87, His89, Cys95 or Glu97 in NikR to alanine eliminates high-affinity nickel binding and abolishes DNA binding but maintains stable protein folding . An unanticipated feature of the NikR structure is that the nickel coordination responds to DNA binding . A six-coordinate nickel site composed of O- or N-donor ligands, but lacking cysteine, forms when NikR binds to operator DNA . Because nickel binding and DNA binding are mediated by different domains within NikR, a communication link between the two domains is implicated, consistent with the finding that the nickel-binding site in a fragment corresponding to the C-terminal domain of NikR is structurally distinct from that found in holo-NikR.

Gut, 2003 Feb, 52(2), 224 - 30
Glutamine deprivation facilitates tumour necrosis factor induced bacterial translocation in Caco-2 cells by depletion of enterocyte fuel substrate; Clark EC et al.; BACKGROUND AND AIMS: Factors that induce luminal bacteria to cross the intestinal epithelium following injury remain poorly defined . The aim of this study was to investigate the interaction between glutamine metabolism, energy supply, and inflammatory mediators in determining the translocation of non-pathogenic bacteria across cultured enterocytes . METHODS: The effect of tumour necrosis factor alpha (TNF-alpha) on translocation of Escherichia coli C25 across Caco-2 epithelial monolayers was studied in the presence of products and inhibitors of glutamine metabolism . Simultaneous measurements of transepithelial electrical resistance (TEER) and flux of lucifer yellow were used to assess effects on the paracellular pathway . Lactate dehydrogenase release was used to monitor enterocyte integrity . Imaging of monolayers in these experimental conditions was undertaken with transmission electron microscopy . RESULTS: Exposure to basolateral TNF-alpha (20 ng/ml) for six hours induced translocation of E coli across Caco-2 but only if accompanied by simultaneous glutamine depletion (p<0.01) . Translocation was inhibited by addition of glutamine for two hours (p<0.01) but not by an isonitrogenous mixture of non-glutamine containing amino acids . Inhibition of glutamine conversion to alpha-ketoglutarate, but not blockade of glutathione or polyamine synthesis, also induced translocation in the presence of TNF-alpha . Manipulations that induced bacterial translocation were associated with a marked reduction in enterocyte ATP levels . No effect of these treatments on paracellular permeability or lactate dehydrogenase release was observed . Conditions in which translocation occurred were associated with the presence of bacteria within enterocyte vacuoles but not the paracellular space . CONCLUSIONS: In inflammatory conditions, the availability of glutamine as an enterocyte fuel substrate is essential for the preservation of a functional barrier to microorganisms . In conditions of acute glutamine depletion, cytokine mediated bacterial translocation appears to be primarily a transcellular process.

Vet Parasitol, 2003 Jan 20, 111(1), 19 - 30
In vitro growth requirements for the fish pathogen Philasterides dicentrarchi (Ciliophora, Scuticociliatida); Iglesias R et al.; Philasterides dicentrarchi is a scuticociliate causing fatal disease in farmed turbot and sea bass . In view of its high virulence and endoparasitic location, this parasite cannot be effectively controlled by formalin baths, and no systemic chemotherapeutic treatments have yet proved effective; immunoprophylaxis may thus be an attractive alternative approach . Since vaccine development is greatly facilitated by axenic culture of the pathogen, we have developed a simple axenic culture system based on commercially available Leibovitz L-15 medium, supplemented with fetal bovine serum, lipids (lecithin and Tween 80), nucleosides and glucose . After 1 week's culture under optimal conditions (salinity 10 per thousand, pH 7.2, temperature between 18 and 23 degrees C), yields of 1-2 x 10(5)cells/ml were obtained . Even cultures with seeding densities as low as 20 cells/ml were found to produce a good yield of ciliates (about 6 x 10(4)cells/ml) after 11 days of incubation . The ciliates thus obtained were free of contamination by other microorganisms, enabling preparation of pure P . dicentrarchi antigens for vaccine development studies.

Water Sci Technol, 2002, 46(11-12), 381 - 7
The quantification and characterization of endocrine disruptor bisphenol-A leaching from epoxy resin; Bae B et al.; Bisphenol-A (BPA), a known endocrine disruptor, is a main building block of epoxy resin which has been widely used as a surface coating agent on residential water storage tanks . Therefore, BPA leaching from the epoxy resin can adversely affect human health . In this study, BPA leaching from three epoxy resins were quantified at 20, 50, 75 and 100 degrees C both in deionized water and the specified test water, respectively . BPA leached to the test water was identified using GC-MS and quantified with GC-FID after a sequential extraction and concentration . The results showed that BPA leaching has occurred in all three samples tested . The quantity of BPA from unit area of epoxy resin coating was in the range of 01.68-273 . 12 microg/m2 for sample A, 29.74-1734.05 microg/m2 for sample B and 52.86-548.78 microg/m2 for sample C depending on the test temperature, respectively . In general, the amount of BPA leashing increased as the water temperature increases . This result implies a higher risk of BPA leaching to drinking water during a summer season . In addition, microbial growth, measured by colony forming units, in epoxy coated water tanks was higher than that in a stainless steel tank . The results suggest that compounds leaching from epoxy resin may support the growth of microorganisms in a residential water holding tank.

Ugeskr Laeger, 2002 Dec 2, 164(49), 5769 - 72
{Lactic bacteria and other probiotics in infections and inflammatory diseases in children . What do we believe? - What do we know?}; Nielsen VR et al.; Probiotics are microorganisms which, when ingested, may have a positive effect in the prevention or treatment of a specific pathologic condition . Probiotics are used for a number of gastrointestinal and certain systemic infectious and inflammatory diseases . The efficacy of selected probiotics to promote recovery from acute viral gastroenteritis in young children is well-documented . However, the role of probiotics in systemic diseases, e.g . atopic dermatitis, need to be confirmed in larger randomised controlled trials . This paper describes the theoretical background for the use of probiotics and reviews results from randomised controlled trials describing the effect of probiotics, particularly in paediatric diseases.

Intern Med, 2002 Dec, 41(12), 1179 - 82
Humidifier lung: possible contribution of endotoxin-induced lung injury; Ohnishi H et al.; A 56-year-old man was admitted with cough, fever, myalgia, and arthralgia . Chest computed tomography demonstrated bilateral diffuse ground-glass opacities predominantly in the upper lungs . Subpleural non-segmental consolidation was observed in the late phase . Hypersensitivity pneumonitis was suspected, and an environmental provocation test with the incidental use of a home ultrasonic humidifier was positive . Unlike typical hypersensitivity pneumonitis, serum KL-6 levels were normal . Although several microorganisms were isolated from the humidifier water, there was no evidence for immune sensitization . We detected high amounts of endotoxin in the humidifier water, which may have contributed to the lung injury of this patient.

C R Biol, 2002 Dec, 325(12), 1159 - 74
{Hemoglobin, from microorganisms to man: a single structural motif, multiple functions}; Wajcman H et al.; Haemoglobins from unicellular organisms, plants or animals, share a common structure, which results from the folding, around the heme group, of a polypeptide chain made from 6-8 helices . Nowadays, deciphering the genome of several species allows one to draw the evolutionary tree of this protein going back to 1800 millions of years, at a time when oxygen began to accumulate in the atmosphere . This permits to follow the evolution of the ancestral gene and of its product . It is likely that, only in complex multicellular species, transport and storage of oxygen became the main physiological function of this molecule . In addition, in unicellular organisms and small invertebrates, it is likely that the main function of this protein was to protect the organism from the toxic effect of O2, CO and NO* . The very high oxygen affinity of these molecules, leading them to act rather as a scavenger as an oxygen carrier, supports this hypothesis . Haemoglobins from microorganisms, which may probably be the closest vestiges to the ancestral molecules, are divided into three families . The first one is made from flavohaemoglobins, a group of chimerical proteins carrying a globin domain and an oxido-reduction FAD-dependant domain . The second corresponds to truncated haemoglobins, which are hexacoordinated with very high oxygen-affinity molecules, 20-40 residues shorter than classical haemoglobins . The third group is made from bacterial haemoglobins such as that of Vitreoscilla . Some specific structural arrangements in the region surrounding the heme are cause of their high oxygen affinity . In plants, two types of haemoglobins are present (non-symbiotic and symbiotic), that arose from duplication of an ancestral vegetal gene . Non-symbiotic haemoglobins, which are probably the oldest, are scarcely distributed within tissues having high energetic consumption . Conversely, symbiotic haemoglobins (also named leghaemoglobins) are present at a high concentration (mM) mostly in the rhizomes of legumes, where they are involved in nitrogen metabolism . In some species, haemoglobin was proposed to be an oxygen sensor bringing to the organism information to adjust metabolism or biosynthesis to the oxygen requirement . Elsewhere haemoglobin may act as final electron acceptors in oxido-reduction pathways . Evolution of haemoglobin in invertebrates followed a large variety of scenarios . Some surprising functions as sulphide acquisition in invertebrates living near hydrothermal vents, or a role in the phototrophism of worm need to be mentioned . In invertebrates, the size of haemoglobin varies from monomers to giant molecules associating up to 144 subunits, while in vertebrates it is always a tetramer . In some species, several haemoglobins, with completely different structure and function, may coexist . This demonstrates how hazardous may be to extrapolate the function of a protein from only structural data.

Circ J, 2003 Jan, 67(1), 26 - 30
Association between hepatitis C virus core protein and carotid atherosclerosis; Ishizaka Y et al.; A link between certain infectious microorganisms and an increased risk of atherosclerotic disease has been suggested . By analyzing the data of subjects who had undergone general health-screening tests, a possible association between carotid atherosclerosis and seropositivity of antibody against hepatitis C virus (HCV) has been previously reported . In the present study, a possible link between carotid atherosclerosis and HCV core protein positivity was assessed, because it is postulated to be a better marker of viremia and thus persistent infection . Of the 1992 enrolled subjects, 496 (25%) had carotid artery plaque, and 25 (1.3%) were positive for HCV core protein . Carotid artery plaque was positive in 480/1967 (24%) and 16/25 (64%) of the core protein-negative and core protein-positive subjects, respectively (p<0.0001 by chi(2) test) . Serum concentrations of transaminases were higher in core protein-positive subjects, but albumin concentrations were not significantly different between the 2 groups . Multivariate logistic regression analysis showed that HCV core protein positivity is an independent predictor of carotid plaque with an odds ratio of 5.61 (95% confidence interval 2.06-15.26, p<0.001) . These data further support the possible link between persistent HCV infection and carotid atherosclerosis in the subjects without severe liver dysfunction.

Nucleic Acids Res, 2003 Jan 1, 31(1), 514 - 6
probeBase: an online resource for rRNA-targeted oligonucleotide probes; Loy A et al.; Ribosomal RNA-(rRNA)-targeted oligonucleotide probes are widely used for culture-independent identification of microorganisms in environmental and clinical samples . ProbeBase is a comprehensive database containing more than 700 published rRNA-targeted oligonucleotide probe sequences (status August 2002) with supporting bibliographic and biological annotation that can be accessed through the internet at Each oligonucleotide probe entry contains information on target organisms, target molecule (small- or large-subunit rRNA) and position, G+C content, predicted melting temperature, molecular weight, necessity of competitor probes, and the reference that originally described the oligonucleotide probe, including a link to the respective abstract at PubMed . In addition, probes successfully used for fluorescence in situ hybridization (FISH) are highlighted and the recommended hybridization conditions are listed . ProbeBase also offers difference alignments for 16S rRNA-targeted probes by using the probe match tool of the ARB software and the latest small-subunit rRNA ARB database (release June 2002) . The option to directly submit probe sequences to the probe match tool of the Ribosomal Database Project II (RDP-II) further allows one to extract supplementary information on probe specificities . The two main features of probeBase, 'search probeBase' and 'find probe set', help researchers to find suitable, published oligonucleotide probes for microorganisms of interest or for rRNA gene sequences submitted by the user . Furthermore, the 'search target site' option provides guidance for the development of new FISH probes.

Saudi Med J, 2002 Dec, 23(12), 1504 - 8
Evaluation of direct light microscopy for rapid detection of microorganisms in blood cultures; Kaplan NM; OBJECTIVE: The aim of this study was to evaluate the use of direct light microscopy (x 40 objective) of unstained uncentrifuged blood-broth mixtures for rapid detection of microorganisms in blood cultures . METHODS: Two thousand two hundred and ninety-four blood cultures were investigated in this study . Blood cultures were processed in the Clinical Microbiology Laboratory, Queen Alia General Hospital, Amman, Jordan from January to December 2001 . We compared the results of direct light microscopy and subcultures in 3 stages: macroscopically positive blood culture bottles, macroscopically negative blood culture bottles after 9-17 hours, and macroscopically negative blood culture bottles after 7 days of incubation . RESULTS: The total positive blood cultures was 434 (18.9%) . Three hundred and fifty three (81.3%) were positive by macroscopic examination and direct light microscopy and grew viable organisms, 34 were macroscopically negative blood cultures that grew viable organisms after 9-17 hours of incubation in which 32 (94.1%) were positive by direct light microscopy and 47 macroscopically negative blood cultures that grew viable organisms after 7 days of incubation in which 45 (95.7%) were positive by direct light microscopy . CONCLUSION: The direct light microscopy methodology proved to be simple, rapid, cost effective, accurate and sensitive technique for the early detection of bacteremia.

J Lab Clin Med, 2003 Jan, 141(1), 33 - 40
Expansion of gammadelta T-cells in Behçet's disease: role of disease activity and microbial flora in oral ulcers; Bank I et al.; Gammadelta T-cells participate in the immune response to infections and in autoimmunity by recognizing bacteria-derived and autologous antigens . The goal of this study was to evaluate the involvement of gammadelta T-cells in Behcet's disease (BD) . Gammadelta T-cells in the peripheral-blood mononuclear cells (PBMCs) of Israeli patients with definite BD (n = 23), normal controls (n = 16), and patients with familial Mediterrranean fever (FMF; n = 20) were evaluated by means of flow cytometry . The responses of patient and control gammadelta T-cells to medium conditioned by microorganisms cultured from an oral ulcer of a patient with active BD were compared . The proportions of CD3(+) and CD8(+) cells in the PBMCs were not significantly different between groups . In contrast, gammadelta-T-cells accounted for 7.01% +/- 4.42% of the PBMCs in BD compared with 3.56% +/- 3.45% in FMF (P < .005) and 3.7% +/- 3.15% in normal individuals (P < .009) . Their numbers were significantly higher during active disease than in remission (9.45% +/- 5.08% versus 2.27% +/- 3.3%; P < .009) . The number of T-cell-receptor gammadelta(+) and Vdelta2(+) cells of BD patients, but not of controls, increased after 96 hours of culture in medium containing supernatant of microorganisms cultured from an oral ulcer in a patient with BD relative to their proportions in control medium: 58.2% vs 13.9% (P < .05) and 28% vs 9% (P < .04), respectively, of the cultured T-cells (n = 4).gammadelta T-cells are expanded in BD PBMCs during active disease . An exaggerated proliferative response to products released by microorganisms present in oral ulcers may play a role in this phenomenon.

J Clin Microbiol, 2003 Jan, 41(1), 324 - 9
Prospective study of the performance of vibrational spectroscopies for rapid identification of bacterial and fungal pathogens recovered from blood cultures; Maquelin K et al.; Rapid identification of microbial pathogens reduces infection-related morbidity and mortality of hospitalized patients . Raman spectra and Fourier transform infrared (IR) spectra constitute highly specific spectroscopic fingerprints of microorganisms by which they can be identified . Little biomass is required, so that spectra of microcolonies can be obtained . A prospective clinical study was carried out in which the causative pathogens of bloodstream infections in hospitalized patients were identified . Reference libraries of Raman and IR spectra of bacterial and yeast pathogens highly prevalent in bloodstream infections were created . They were used to develop identification models based on linear discriminant analysis and artificial neural networks . These models were tested by carrying out vibrational spectroscopic identification in parallel with routine diagnostic phenotypic identification . Whereas routine identification has a typical turnaround time of 1 to 2 days, Raman and IR spectra of microcolonies were collected 6 to 8 h after microbial growth was detected by an automated blood culture system . One hundred fifteen samples were analyzed by Raman spectroscopy, of which 109 contained bacteria and 6 contained yeasts . One hundred twenty-one samples were analyzed by IR spectroscopy . Of these, 114 yielded bacteria and 7 were positive for yeasts . High identification accuracy was achieved in both the Raman (92.2%, 106 of 115) and IR (98.3%, 119 of 121) studies . Vibrational spectroscopic techniques enable simple, rapid, and accurate microbial identification . These advantages can be easily transferred to other applications in diagnostic microbiology, e.g., to accelerate identification of fastidious microorganisms.

J Clin Microbiol, 2003 Jan, 41(1), 310 - 7
Performance evaluation of the VERSANT HCV RNA qualitative assay by using transcription-mediated amplification; Gorrin G et al.; A preclinical evaluation of a qualitative assay for the detection of hepatitis C virus (HCV) RNA by transcription-mediated amplification (TMA) was conducted according to the guidelines of the National Committee for Clinical Laboratory Standards and the U.S . Food and Drug Administration . Our results showed that this assay, HCV TMA, detected 95% of samples with HCV RNA concentrations of 5.3 IU/ml and 29 copies/ml . HCV TMA showed an overall specificity of 99.6% and was highly reproducible, detecting 99.3% of samples with HCV RNA concentrations of 50 copies/ml across seven different lots of reagents . Experiments with clinical samples showed that HCV TMA detected all HCV genotypes with similar efficiencies, detecting > or = 95% of samples at 50 HCV RNA copies/ml from patients infected with HCV genotypes 1a, 2b, 3a, 4a, 5a, and 6a . In experiments with RNA transcripts, HCV TMA detected > or = 96.6% of transcripts derived from HCV genotypes 1a, 1b, 2a, 2c, 3a, 4a, 5a, and 6a at 50 HCV RNA copies/ml . Detection of transcripts derived from HCV genotype 2b was slightly lower (88.4%) at 50 copies/ml but was 97.0% at 75 copies/ml . In addition, HCV TMA exhibited robust performance in detecting HCV RNA in samples subjected to various conditions commonly encountered in a clinical laboratory, including long-term storage, multiple freeze-thaw cycles, different collection tubes, and the presence of endogenous substances, commonly prescribed drugs, or other microorganisms and viruses . With its high sensitivity, specificity, reproducibility, and equivalent genotype reactivity, HCV TMA may provide an attractive alternative for routine qualitative HCV RNA testing in clinical laboratories.

Infect Control Hosp Epidemiol, 2002 Dec, 23(12 Suppl), S3 - 40
Guideline for Hand Hygiene in Health-Care Settings: recommendations of the Healthcare Infection Control Practices Advisory Committee and the HICPAC/SHEA/APIC/IDSA Hand Hygiene Task Force; Boyce JM et al.; The Guideline for Hand Hygiene in Health-Care Settings provides health-care workers (HCWs) with a review of data regarding handwashing and hand antisepsis in health-care settings . In addition, it provides specific recommendations to promote improved hand-hygiene practices and reduce transmission of pathogenic microorganisms to patients and personnel in health-care settings . This report reviews studies published since the 1985 CDC guideline (Garner JS, Favero MS . CDC guideline for handwashing and hospital environmental control, 1985 . Infect Control 1986;7:231-43) and the 1995 APIC guideline (Larson EL, APIC Guidelines Committee . APIC guideline for handwashing and hand antisepsis in health care settings . Am J Infect Control 1995;23:251-69) were issued and provides an in-depth review of hand-hygiene practices of HCWs, levels of adherence of personnel to recommended handwashing practices, and factors adversely affecting adherence . New studies of the in vivo efficacy of alcohol-based hand rubs and the low incidence of dermatitis associated with their use are reviewed . Recent studies demonstrating the value of multidisciplinary hand-hygiene promotion programs and the potential role of alcohol-based hand rubs in improving hand-hygiene practices are summarized . Recommendations concerning related issues (e.g., the use of surgical hand antiseptics, hand lotions or creams, and wearing of artificial fingernails) are also included.

Nat Biotechnol, 2003 Feb, 21(2), 143 - 9 Epub 2003 Jan 06.
Total biosynthesis of hydrocortisone from a simple carbon source in yeast; Szczebara FM et al.; We report on the production of hydrocortisone, the major adrenal glucocorticoid of mammals and an important intermediate of steroidal drug synthesis, from a simple carbon source by recombinant Saccharomyces cerevisiae strains . An artificial and fully self-sufficient biosynthetic pathway involving 13 engineered genes was assembled and expressed in a single yeast strain . Endogenous sterol biosynthesis was rerouted to produce compatible sterols to serve as substrates for the heterologous part of the pathway . Biosynthesis involves eight mammalian proteins (mature forms of CYP11A1, adrenodoxin (ADX), and adrenodoxin reductase (ADR); mitochondrial forms of ADX and CYP11B1; 3beta-HSD, CYP17A1, and CYP21A1) . Optimization involved modulating the two mitochondrial systems and disrupting of unwanted side reactions associated with ATF2, GCY1, and YPR1 gene products . Hydrocortisone was the major steroid produced . This work demonstrates the feasibility of transfering a complex biosynthetic pathway from higher eukaryotes into microorganisms.

Appl Environ Microbiol, 2003 Jan, 69(1), 452 - 60
Compost-induced suppression of Pythium damping-off is mediated by fatty-acid-metabolizing seed-colonizing microbial communities; McKellar ME et al.; Leaf composts were studied for their suppressive effects on Pythium ultimum sporangium germination, cottonseed colonization, and the severity of Pythium damping-off of cotton . A focus of the work was to assess the role of fatty-acid-metabolizing microbial communities in disease suppression . Suppressiveness was expressed within the first few hours of seed germination as revealed by reduced P . ultimum sporangium germination, reduced seed colonization, and reduced damping-off in transplant experiments . These reductions were not observed when cottonseeds were sown in a conducive leaf compost . Microbial consortia recovered from the surface of cottonseeds during the first few hours of germination in suppressive compost (suppressive consortia) induced significant levels of damping-off suppression, whereas no suppression was induced by microbial consortia recovered from cottonseeds germinated in conducive compost (conducive consortia) . Suppressive consortia rapidly metabolized linoleic acid, whereas conducive consortia did not . Furthermore, populations of fatty-acid-metabolizing bacteria and actinobacteria were higher in suppressive consortia than in conducive consortia . Individual bacterial isolates varied in their ability to metabolize linoleic acid and protect seedlings from damping-off . Results indicate that communities of compost-inhabiting microorganisms colonizing cottonseeds within the first few hours after sowing in a Pythium-suppressive compost play a major role in the suppression of P . ultimum sporangium germination, seed colonization, and damping-off . Results further indicate that fatty acid metabolism by these seed-colonizing bacterial consortia can explain the Pythium suppression observed.

Appl Environ Microbiol, 2003 Jan, 69(1), 320 - 6
Evaluation of PCR amplification bias by terminal restriction fragment length polymorphism analysis of small-subunit rRNA and mcrA genes by using defined template mixtures of methanogenic pure cultures and soil DNA extracts; Lueders T et al.; Terminal restriction fragment length polymorphism (T-RFLP) analysis is a widely used method for profiling microbial community structure in different habitats by targeting small-subunit (SSU) rRNA and also functional marker genes . It is not known, however, whether relative gene frequencies of individual community members are adequately represented in post-PCR amplicon frequencies as shown by T-RFLP . In this study, precisely defined artificial template mixtures containing genomic DNA of four different methanogens in various ratios were prepared for subsequent T-RFLP analysis . PCR amplicons were generated from defined mixtures targeting not only the SSU rRNA but also the methyl-coenzyme M reductase (mcrA/mrtA) genes of methanogens . Relative amplicon frequencies of microorganisms were quantified by comparing fluorescence intensities of characteristic terminal restriction fragments . SSU ribosomal DNA (rDNA) template ratios in defined template mixtures of the four-membered community were recovered absolutely by PCR-T-RFLP analysis, which demonstrates that the T-RFLP analysis evaluated can give a quantitative view of the template pool . SSU rDNA-targeted T-RFLP analysis of a natural community was found to be highly reproducible, independent of PCR annealing temperature, and unaffected by increasing PCR cycle numbers . Ratios of mcrA-targeted T-RFLP analysis were biased, most likely by PCR selection due to the degeneracy of the primers used . Consequently, for microbial community analyses, each primer system used should be evaluated carefully for possible PCR bias . In fact, such bias can be detected by using T-RFLP analysis as a tool for the precise quantification of the PCR product pool.

Tohoku J Exp Med, 2002 Oct, 198(2), 125 - 32
Sulfite is generated from PAPS by activated neutrophils; Mitsuhashi H et al.; We previously reported that neutrophils produce sulfite in response to stimulation with lipopolysaccharide, and sulfite production is dependent on inorganic sulfate contained in culture media . Microorganisms such as yeast assimilate sulfate, during which process sulfite is generated by reduction of 3'-phosphoadenosine 5'-phosphosulfate (PAPS), an activated sulfate donor . However, little is known about how sulfite is produced in mammalian cells . In the current study, we demonstrated that chlorate, a specific inhibitor for PAPS synthesis, significantly suppressed production of sulfite by activated neutrophils obtained from rat peritoneal cavity that had been injected with glycogen to induce inflammation . Addition of excess amounts of PAPS could partially overcome the inhibitory effect of chlorate . Moreover, sulfite production from PAPS was clearly demonstrated in the cytosolic fraction of activated neutrophils . These findings strongly suggest that sulfite is generated, at least in part, from PAPS by activated neutrophils.

Vet Clin North Am Small Anim Pract, 2003 Jan, 33(1), 89 - 118
Abdominal, thoracic, and pericardial effusions; Alleman AR; The laboratory evaluation of abdominal, thoracic, and pericardial effusions is a useful diagnostic tool for the assessment of disease states that result in fluid accumulation . Although the numeric values pertaining to cell count and protein content are important, the microscopic evaluation is a critical aspect of the diagnostic procedure; not only does it allow complete classification of the fluid but it allows identification of specific cell types or microorganisms that might be responsible for the fluid accumulation . These findings should always be interpreted in conjunction with the history, signalment, physical findings, and other diagnostic aids in making a definitive diagnosis.

J Clin Invest, 2003 Jan, 111(1), 109 - 19
src homology 2 domain-containing tyrosine phosphatase SHP-1 controls the development of allergic airway inflammation; Kamata T et al.; Th2 cells are generated from naive CD4 T cells upon T cell receptor (TCR) recognition of antigen and IL-4 stimulation and play crucial roles in humoral immunity against infectious microorganisms and the pathogenesis of allergic and autoimmune diseases . A tyrosine phosphatase, SHP-1, that contains src homology 2 (SH2) domains is recognized as a negative regulator for various intracellular signaling molecules, including those downstream of the TCR and the IL-4 receptor . Here we assessed the role of SHP-1 in Th1/Th2 cell differentiation and in the development of Th2-dependent allergic airway inflammation by using a natural SHP-1 mutant, the motheaten mouse . CD4 T cells appear to develop normally in the heterozygous motheaten (me/+) thymus even though they express decreased amounts of SHP-1 (about one-third the level of wild-type thymus) . The me/+ naive splenic CD4 T cells showed enhanced activation by IL-4 receptor-mediated signaling but only marginal enhancement of TCR-mediated signaling . Interestingly, the generation of Th2 cells was increased and specific cytokine production of mast cells was enhanced in me/+ mice . In an OVA-induced allergic airway inflammation model, eosinophilic inflammation, mucus hyperproduction, and airway hyperresponsiveness were enhanced in me/+ mice . Thus, SHP-1 may have a role as a negative regulator in the development of allergic responses, such as allergic asthma.

Aust Endod J, 2002 Dec, 28(3), 112 - 5
Discuss that the coronal seal is more important than the apical seal for endodontic success; Sritharan A; One of the main principles for successful root canal treatment is the prevention of microorganisms and toxins from the oral flora penetrating through the root canal system into the periapical tissues (1) . This is achieved by obturating the root canal system completely, including the coronal and apical ends . Inadequate obturation of the root canal system has been found to be the most frequent cause of failure after endodontic treatment (2) . A number of studies have indicated that leakage, whether from a coronal or apical direction, adversely affects the success of root canal treatment (3-7) . The significance of the coronal seal has been increasingly recognized in the dental literature (4, 5, 8, 9) and in more recent times, it has been suggested that apical leakage may not be the most important factor leading to the failure of endodontic treatment--but that coronal leakage is far more likely to be the major determinant of clinical success or failure (10) . This paper will discuss aspects of: endodontic success and failure; the use of leakage studies to assess the quality of the seal; the significance of both apical and coronal leakage; followed by a review of the literature.

Protein Expr Purif, 2003 Jan, 27(1), 19 - 26
Expression and purification of functional, recombinant Trypanosoma cruzi complement regulatory protein; Beucher M et al.; The complement regulatory protein (CRP) of Trypanosoma cruzi is a developmentally regulated glycosylphosphatidylinositol (GPI)-anchored membrane protein that protects the parasite from complement-mediated killing, and is an important virulence determinant of the microorganism . CRP binds human complement components C3b and C4b to restrict activation of the complement cascade . Here, we report production of functional, recombinant T . cruzi CRP in mammalian cells and a one-step purification of the recombinant protein . Exchange of the crp DNA sequence encoding the carboxy-terminal GPI signal sequence with the corresponding sequence of decay accelerating factor (DAF) was necessary for recognition, cleavage, and addition of GPI in mammalian cells . CRP production was assessed in two mammalian cell lines with crp-daf gene expression driven by three different transcription control regions: Rous sarcoma virus long terminal repeat, cytomegalovirus (CMV) immediate early gene, and chicken beta-actin promoter/CMV enhancer . We present evidence that CRP produced in transfected Chinese hamster Ovary (CHO) cells was functional and protected the cells from complement-mediated lysis . To facilitate purification of the recombinant protein, a hexahistidyl tag was incorporated at 3(') end of the cDNA upstream of the GPI anchor addition sequence . An additional histidine fusion construct was made that allowed for secretion and recovery of recombinant protein from culture supernatant fluid . Both membrane and secreted forms of the protein were purified in one step by nickel nitrilotriacetic acid . The production and purification of functionally active CRP in a non-infectious expression system will allow for structure and function studies aimed at identifying the active site(s) of this protein.

J Law Med Ethics, 2002 Fall, 30(3 Suppl), 96 - 104
Protecting our vulnerable food supply; Pestronk RM et al.; From farmyard to dinner table, our food supply presents ample opportunity for dangerous microorganisms or their products to thrive and infect or intoxicate human beings, often with harmful and sometimes fatal results . Traditional controls to protect the food supply include, but are not limited to, law and regulation . But law and regulation are only enablers, an underpinning . Most important to protection of the food supply is organizational leadership and commitment at federal, state, and local levels of government to protect the public's health . This article provides examples of such leadership in locales as diverse as Australia, New Zealand, and Genesee County, Michigan . Even when a supportive law is in place and the will and resources to make the law work exist, competing political and economic world-views are constantly at work to amend the law and thus adversely affect the public's health.

Vet Immunol Immunopathol, 2003 Jan 10, 91(1), 1 - 12
TOLL-like receptors linking innate and adaptive immune response; Werling D et al.; Invading pathogens are controlled by the innate and adaptive arms of the immune system . Adaptive immunity, which is mediated by B and T lymphocytes, recognises pathogens by rearranged high affinity receptors . However, the establishment of adaptive immunity is often not rapid enough to eradicate microorganisms as it involves cell proliferation, gene activation and protein synthesis . More rapid defense mechanisms are provided by innate immunity, which recognises invading pathogens by germ-line-encoded pattern recognition receptors (PRR) . Recent evidence shows that this recognition can mainly be attributed to the family of TOLL-like receptors (TLR) . Binding of pathogen-associated molecular patterns (PAMP) to TLR induces the production of reactive oxygen and nitrogen intermediates (ROI and RNI), pro-inflammatory cytokines, and up-regulates expression of co-stimulatory molecules, subsequently initiating the adaptive immunity . In this review, we will summarize the discovery and the critical roles of the TLR family in host defense, briefly allude to signaling mechanisms mediating the response to TLR ligands, and will provide an update on current knowledge regarding the ligand specificity of these receptors and their role in immunity of domestic animals, particularly cattle.

Zh Mikrobiol Epidemiol Immunobiol, 2002 Nov-Dec, (6), 105 - 11
{Flow laser cytometry for evaluation of the human immune system}; Pinegin BV et al.; Some flow laser cytometry (FLC) techniques intended for studies of the immune system cells are reviewed . A widespread analytical method is the phenotyping of lymphocytes by the markers they express . The use of FLC permits the evaluation of practically all functional parameters of immunocompetent cells . Thus, to analyze their ingestive and microbicidal activity fluorochrome-labeled microorganisms are used . The apploication of indicator dyes makes it possible to evaluate calcium mobilization and formation of active forms of oxygen . FLC is used for the identification of cytokines inside the cell and in the medium . The authors propose tests for the analysis of the proliferative activity of lymphocytes, the cytotoxicity of natural killers, the evaluation of apoptosis and protein processing with monocytes/macrophages.

Vet J, 2002 Nov, 164(3), 275 - 9
Influence of diet on the shedding of Candida glabrata by experimentally infected preweaned calves; Elad D et al.; The influence of three diets, comprising dam's milk (DM) from the same farm, commercial milk replacer with (CMRL) or without 3.2% lactose (CMR), on the duration and intensity of Candida glabrata shedding in the faeces of preweaned calves following experimental oral infection was examined . Shedding of other potential enteric pathogens was also monitored . The duration and intensity of C . glabrata shedding in DM-fed calves were reduced significantly compared with the calves fed the two diets based on milk replacers . Consequently, feeding calves with DM might disrupt the infective cycle, resulting in the yeast's elimination from a farm . In the CMR and CMRL groups, the periods of intensive shedding of C . glabrata and rotavirus overlapped but no diarrhoea was associated with the shedding of either microorganism . There was no evidence that lactose diminished colonization in vivo .

Vet J, 2002 Nov, 164(3), 202 - 15
Recent developments in avian influenza research: epidemiology and immunoprophylaxis; Tollis M et al.; Influenza A viruses have been isolated from humans, from several other mammalian species and a wide variety of avian species, among which, wild aquatic birds represent the natural hosts of influenza viruses . The majority of the possible combinations of the 15 haemagglutinin (HA) and nine neuraminidase (NA) subtypes recognized have been identified in isolates from domestic and wild birds . Infection of birds can cause a wide range of clinical signs, which may vary according to the host, the virus strain, the host's immune status, the presence of any secondary exacerbating microorganisms and environmental factors . Most infections are inapparent, especially in waterfowl and other wild birds . In contrast, infections caused by viruses of H5 and H7 subtypes can be responsible for devastating epidemics in poultry . Despite the warnings to the poultry industry about these viruses, in 1997 an avian H5N1 influenza virus was directly transmitted from birds to humans in Hong Kong and resulted in 18 confirmed infections, thus strengthening the pandemic threat posed by avian influenza (AI) . Indeed, reassortant viruses, harbouring a combination of avian and human viral genomes, have been responsible for major pandemics of human influenza . These considerations warrant the need to continue and broaden efforts in the surveillance of AI . Control programmes have varied from no intervention, as in the case of the occurrence of low pathogenic (LP) AI (LPAI) viruses, to extreme, expensive total quarantine-slaughter programmes carried out to eradicate highly pathogenic (HP) AI (HPAI) viruses . The adoption of a vaccination policy, targeted either to control or to prevent infection in poultry, is generally banned or discouraged . Nevertheless, the need to boost eradication efforts in order to limit further spread of infection and avoid heavy economic losses, and advances in modern vaccine technologies, have prompted a re-evaluation of the potential use of vaccination in poultry as an additional tool in comprehensive disease control strategies . This review presents a synthesis of the most recent research on AI that has contributed to a better understanding of the ecology of the virus and to the development of safe and efficacious vaccines for poultry .

Ann Pharmacother, 2003 Jan, 37(1), 132 - 5
Bioterrorism web sites for pharmacists; Misita CP et al.; OBJECTIVE: To identify Internet Web sites for ease of accessibility to bioterrorism-related information, comprehensive provision of bioterrorism-related information, and provision of bioterrorism information that specifically pertains to the pharmacy profession . DATA SOURCES: Web sites of national pharmacy organizations, US government agencies, and medical organizations, as well as Web sites related to bioterrorism . DATA SYNTHESIS: Pharmacists need access to relevant bioterrorism information in a timely manner . An evaluation of Web sites was performed to identify those that include a discussion of the potential infectious microorganisms and prevention and treatment methods, as well as unique features for pharmacy practice . RESULTS: The American Society of Health-System Pharmacists and American Pharmaceutical Association Web sites provide pharmacy-specific recommendations . The Centers for Disease Control and Prevention provides biological agent information and health department contact numbers . Additional agent-specific data are provided by the American Medical Association, The Johns Hopkins University, and the Food and Drug Administration (FDA) Web sites . Information addressing food safety is provided by the FDA . CONCLUSIONS: Pharmacy-specific bioterrorism information is available only at selected national pharmacy organization Internet Web sites . However, other Web sites provide comprehensive bioterrorism information useful for pharmacists.

Folia Microbiol (Praha), 2002, 47(5), 521 - 6
The effect of fulvic acids on the toxicity of lead and manganese to arbuscular mycorrhizal fungus Glomus intraradices; Malcova R et al.; The effect of fulvic acids (FA) on arbuscular mycorrhizal (AM) fungi and on the toxicity of lead and manganese toward these symbionts were demonstrated in vitro . Incubation of root segments colonized with the AM fungus Glomus intraradices in undiluted fraction of FA (813 mg/L carbon) decreased an outgrowth of intraradical hyphae . Diluted FA solutions (< 271 mg/L C) did not influence the proportion of root segments bearing proliferating hyphae; solution containing 27.1 mg/L C even increased the proliferation . A decrease of heavy metal toxicity toward the fungus was observed when FA (81.3 mg/L C) were added to the solutions containing higher concentrations (< or = 100 mumol/L) of Mn and Pb; the positive effect of FA was not significant at higher concentrations of metals (0.5 mmol/L) . A short-term cultivation of six different saprophytic microorganisms--three actinomycetes and three filamentous fungi--on the FA media (81.3 mg/L C) did not result in a modification of hyphal proliferation from the root segments subsequently incubated in these media.

J Virol, 2003 Jan, 77(2), 1347 - 56
Contrasting roles of endosomal pH and the cytoskeleton in infection of human glial cells by JC virus and simian virus 40; Ashok A et al.; Infection of eukaryotic cells by pathogens requires the efficient use of host cell endocytic and cytoplasmic transport mechanisms . Understanding how these cellular functions are exploited by microorganisms allows us to better define the basic biology of pathogenesis while providing better insight into normal cellular functions . In this report we compare and contrast intracellular transport and trafficking of the human polyomavirus JC virus (JCV) with that of simian virus 40 (SV40) . We have previously shown that infection of human glial cells by JCV requires clathrin-dependent endocytosis . In contrast, infection of cells by SV40 proceeds by caveola-dependent endocytosis . We now examine the roles of endosomal pH and the cellular cytoskeleton during infection of glial cells by both viruses . Our results demonstrate that JCV infection is sensitive to disruption of endosomal pH, whereas SV40 infection is pH independent . Infection by JCV is inhibited by treatment of glial cells with cytochalasin D, nocodazole, and acrylamide, whereas SV40 infection is affected only by nocodazole . These data point to critical differences between JCV and SV40 in terms of endocytosis and intracellular trafficking of their DNA genomes to the nucleus . These data also suggest a unique sequential involvement of cytoskeletal elements during infection of glial cells by JCV.

J Gen Appl Microbiol, 1999 Aug, 45(4), 143 - 147
Tryptophan biosynthesis and production of other related compounds from indolepyruvic acid by mixed ruminal bacteria, protozoa, and their mixture in vitro; Mohammed N et al.; Tryptophan (Trp) biosynthesis and the production of other related compounds by mixed ruminal bacteria (B), protozoa (P), and their mixture (BP) in an in vitro system were quantitatively investigated by using 1 mM of indole-3-pyruvic acid (IPA) as substrate . Ruminal microorganisms were anaerobically incubated at 39 degrees C for 12 h . Trp and other related compounds in both the supernatants and the microbial hydrolyzates of the incubation were analyzed by HPLC . As a whole, about 334, 440, and 436 &mgr;M of Trp were produced from IPA in 12 h by B, P, and BP suspensions, respectively . In the B suspension, a greater portion of synthesized Trp (242 &mgr;M) from IPA was accumulated as free form in the medium, whereas a large amount of Trp (92 &mgr;M) was incorporated into cell protein in a 12-h incubation . On the other hand, in the P suspension, a large amount of Trp (475 &mgr;M) from IPA was also found as free form in the supernatant in a 12-h incubation . Protozoa did not incorporate Trp into cell protein, but they liberated endogenous Trp (34 &mgr;M) into the medium . The net productions of Trp from IPA were 344.3 and 447.7 &mgr;mol/g of microbial nitrogen in 12 h by B and P suspensions, respectively . The ability of P to synthesize Trp from IPA was about 30% higher than that of B in 12 h . Trp produced from IPA by B, P, and BP suspensions were simultaneously degraded into its related compounds, and among them, indoleacetic acid (IAA) was a major product found in all microbial suspensions . Productions of IAA were 124, 25, and 99 &mgr;M from IPA in 12 h by B, P, and BP suspensions, respectively . The formation of indolelactic acid (ILA) from IPA was observed for the first time in all microbial suspensions, and it was about 84, 24, and 54 &mgr;M in 12 h by B, P, and BP, respectively . Higher IAA and ILA productions were observed in B when compared with P . A small amount of skatole (SKT) (26 &mgr;M) was produced from IPA in B, whereas a sizable amount of SKT (38 &mgr;M) was found in BP after a 12-h incubation . p-Cresol (CRL) was also produced from IPA by both B (43 &mgr;M) and BP (65 &mgr;M) suspensions in 12 h, and this is also the first discovery to show the formation of CRL from IPA by B and BP suspensions . BP suspension was more active to produce both SKT and CRL from IPA, though P suspension has no ability to produce either SKT or CRL from IPA during a 12-h incubation.

J Gen Appl Microbiol, 1999 Oct, 45(5), 247 - 251
Isolation of a microorganism capable of degrading poly-(L-lactide); Ikura Y et al.; The isolation of poly-(L-lactide) (PLA)-degrading microorganisms was investigated . A PLA-degrading actinomycete, strain No . 3118, was isolated and tentatively identified as a member of the genus Amycolatopsis . The optimum conditions for degradation of PLA were 43 degrees C at about pH 7 in a mineral salt medium with a low concentration of organic nutrients (0.002% yeast extract) . The original shape of PLA film (Mw=2.3x10(5) after sterilization, 20 &mgr;m thick) disappeared within 2 weeks . Lactic acid was detected after the film was incubated with culture supernatant.

J Gen Appl Microbiol, 1997 Feb, 43(1), 9 - 15
Synthesis of phenylalanine and production of other related compounds from phenylpyruvic acid and phenylacetic acid by ruminal bacteria, protozoa, and their mixture in vitro; Amin MR et al.; Phenylalanine (Phe) synthesis and the production of other related compounds by mixed ruminal bacteria (B), protozoa (P), and a combination of the two mixture (BP) in an in vitro system were quantitatively investigated using phenylpyruvic acid (PPY) and phenylacetic acid (PAA) as substrates . Rumen microorganisms were collected from fistulated goats fed lucerne cubes (Medicago sativa) and a concentrated mixture twice a day . Microbial suspensions were anaerobically incubated at 39 degrees C for 12 h . Phe and some other related compounds in both supernatants and microbial hydrolysates of the incubations were analysed by HPLC . A large quantity of Phe was produced from both PPY and PAA not only in B but also in P . In B suspensions, free Phe also accumulated in the medium only when PPY was used as a substrate . The ability of B to synthesize Phe from both PPY and PAA (expressed as unit 'per microbial nitrogen') was 5.1 and 24.8% higher than P, respectively . Phe production from PPY in B and P was 43.5 and 55.2% higher than that from PAA . Large amounts of PAA (17-27%) were produced from PPY in all microbial suspension and production amounts were similar in B and P . Small amounts of benzoic acid (BZA) were produced from PPY and PAA in B, P, and BP, and higher BZA production was observed in P as compared to B . Phenylpropionic acid (PPR) was produced in B from both PPY and PAA, but not in P or BP . A trace amount of phenyllactic acid (PLA) was detected only from PPY in B . Higher concentrations of an unknown compound from PPY and PAA were found to be accumulated in the body protein of B and also in the medium of P, and production of the compound from both PPY and PAA was also higher in B than P.

J Gen Appl Microbiol, 1997 Feb, 43(1), 1 - 7
In vitro metabolism of phenylalanine by ruminal bacteria, protozoa, and their mixture; Amin MR et al.; An in vitro study was conducted to examine the metabolism of phenylalanine (Phe) by mixed rumen bacteria (B), mixed rumen protozoa (P), and a combination of the two (BP) . Rumen microorganisms were collected from fistulated goats fed lucerne cubes (Medicago sativa) and a concentrated mixture twice a day . Microbial suspensions were anaerobically incubated at 39 degrees C for 12 h . Phe and some other related compounds in both supernatants and microbial hydrolysates of the incubations were analysed by HPLC . The net degradation rate (&mgr;mol/g microbial nitrogen) of Phe in B was about 1.5-fold higher than that in P . Phe was converted mainly into phenylacetic acid (PAA) and unknown compound(s) that presumably involved tyrosine in B, P, and BP during the 12 h incubation period . Small amounts of benzoic acid (BZA), and traces of phenylpropionic acid (PPR) and phenyllactic acid (PLA) were also produced from Phe . PAA production in B was found to be higher than that in P, whereas it was significantly higher in BP . Although BZA production was less than one-tenth that of PAA production, it was higher in P than in B and BP . PPR was detected in both B and BP, but not in P . PLA was detected only in B . The production of unknown compound(s) was higher in B than in P and BP.

J Gen Appl Microbiol, 1997 Apr, 43(2), 81 - 87
Evidence for existence of lectins on the ruminal bacteria from steers fed roughage and concentrate diets; Meng Q et al.; Mixed rumen bacteria, isolated by centrifugation from the rumen of steers fed a roughage (R) or concentrate (C) diet, were used to determine if lectins are present on rumen bacteria, based on hemagglutination (HA) and HA inhibition assays in vitro . Rumen bacteria from steers fed either diet agglutinated erythrocytes from cattle, sheep, pigs, and rats, suggested that lectins exist on rumen bacteria . Bacterial HA titers from steers receiving the R diet were much higher (p<0.001) than those from steers fed the C diet, depending on the erythrocyte source used . Centrifugation at 20,000xg for 30 min fractionated the rumen bacteria into upper (U) and lower (L) layers . The HA titers of the U bacterial fractions were significantly higher (p<0.001) than those of the L fractions . A remarkable reduction or complete disappearance of HA titers following treatment of rumen bacteria with protease, trypsin, sodium dodecyl sulfate (SDS) or sodium periodate indicates that rumen bacterial lectins are probably glycoproteins . Lectin specificity for saccharides (galactose, lactose, N-acetyl-D-galactosamine, methyl-alpha-D-galactopyranoside and methyl-beta-D-galactopyranoside) and glycoproteins (mucin, fetuin, and thyroglobulin) was found in the RU, RL, and CU bacterial fractions; no specific binding was determined in the CL fractions . The potential role of lectins in mediating the attachment of rumen bacteria to feed particles, rumen epithelia and other microorganisms is discussed.

J Gen Appl Microbiol, 1997 Aug, 43(4), 217 - 224
Isolation and characterization of Saccharomyces cerevisiae genes differentially expressed under different growth conditions; Wang WY et al.; The budding yeast Saccharomyces cerevisiae, like many other microorganisms, responds to nutrient starvation by arresting growth and entering into a non-proliferating stationary phase . Studies on the response of S . cerevisiae cells to growth arrest might provide further insight into the non-proliferative states of cells in multi-cellular eukaryotic organisms . Changes might occur at the transcription, translation, and post-translational levels in cells upon entry into the stationary phase . To search for the genes differentially expressed in yeast cells during different growth phases, we have performed systematic Northern hybridization experiments using probes prepared for a large number of genes/ORFs . We have thus isolated and characterized 42 cDNA clones containing genes hyper-expressed in the post-diauxic phase . Some of them have already been characterized, and many others show similarity to known yeast genes or genes of other organisms . However, eleven of them were found to be unrelated to any known genes . We have characterized some of these genes as described below . Also, a possible cis-element for transcriptional regulation was identified.

J Gen Appl Microbiol, 1998 Feb, 44(1), 1 - 9
Effects of salinomycin and vitamin B(6) on in vitro metabolism of phenylalanine and its related compounds by ruminal bacteria, protozoa and their mixture; Amin MR et al.; An in vitro study was conducted to examine the effects of salinomycin (SL) and vitamin B(6) (B(6)) on the production of phenylalanine (Phe) from phenylpyruvic acid (PPY) and phenylacetic acid (PAA) and of PAA from Phe and PPY by mixed rumen bacteria (B), mixed rumen protozoa (P) and their mixture (BP) . Rumen microorganisms were collected from fistulated goats fed lucerne cubes (Medicago sativa) and a concentrate mixture (3 : 1) twice a day . Microbial suspensions were anaerobically incubated at 39 degrees C for 12 h . Phe and some other related compounds in both supernatants and microbial hydrolysates of the incubations were analyzed by HPLC . When PPY was used as a substrate, it completely disappeared without additives and converted mainly to Phe and PAA on the average by 396 and 178, 440 and 189, and 439 and 147 &mgr;M in B, P and BP, respectively, during the 12 h incubation period . The rate of disappearance showed no significant differences between the microbial suspensions with and without SL and B(6) during the incubation period . The production of Phe from PPY with SL was enhanced (p<0.05) by 40, 20 and 19% in B, P and BP, respectively, while PAA production from PPY with SL was inhibited (p<0.05) by 35, 37 and 38% in B, P and BP, respectively, during the 12 h incubation period . On the other hand, with B(6), the production of Phe and PAA from PPY tended to be enhanced by 14 and 17, 9 and 11, and 7 and 22% in B, P and BP, respectively, during the 12 h incubation period . When PAA added as a substrate was incubated in the incubation medium without any additives, it disappeared by 483, 462 and 507 &mgr;M and converted mainly to Phe on the average by 231, 244 and 248 &mgr;M in B, P and BP, respectively . The disappearance of PAA with SL was inhibited (p<0.05) by 16, 15 and 20%, in B, P and BP, respectively, whereas the disappearance of PAA with B6 was almost the same as that without B(6) in B and BP suspensions but tended to be enhanced by more than 9% in P suspensions during the 12 h incubation period . The production of Phe from PAA with SL tended to be inhibited by 12, 11 and 8% in B, P and BP, respectively, during the 6 h incubation period, but the inhibition was weakened during the 12 h incubation period, whereas Phe production from PAA with B(6) tended to be enhanced by 13, 16 and 8% in B, P and BP, respectively . When Phe was added as a substrate, the net Phe disappearance without additives was 549, 365 and 842 &mgr;M and converted mainly to PAA on the average by 254, 205 and 461 &mgr;M in B, P and BP, respectively . The net disappearance of Phe with SL was inhibited (p<0.05) by 38, 28 and 46%, whereas the net disappearance of Phe with B(6) was enhanced (p<0.05) by 9, 8 and 7% in B, P and BP, respectively . The production of PAA from Phe with SL was inhibited (p<0.05) by 73, 54 and 76% in B, P and BP, respectively . On the other hand, with B(6), PAA production from Phe was enhanced (p<0.05) by 19, 18 and 20% in B, P and BP, respectively . Based on these results, it seems that SL inhibited Phe disappearance and enhanced the synthesis of Phe from PPY, though not from PAA, and accumulated free Phe in the medium, whereas B(6) also enhanced Phe synthesis both from PPY and PAA, which could provide additional amino N for animals.

J Oral Implantol, 2001, 27(4), 164 - 73
A new implant designed to maximize contact with trabecular bone: survival to 18 months; Morris HF et al.; PROBLEM: A roughened, commercially pure titanium (CP-Ti) implant design has been developed that features a different length and pitch for each screw thread to direct functional stresses away from cortical bone and to the more resilient trabecular bone . Abutment-implant connection is made using a conical taper to provide a seal against invasion by microorganisms . PURPOSE: To assess short-term (18 months) clinical performance of this innovative implant design . METHODS: A total of 1,419 implants were placed in 313 patients to support 419 prostheses in a multidisciplinary, multicentered, prospective clinical study conducted by the Ankylos Implant Clinical Research Group (AICRG) . More than 100 dentists at 32 centers in the United States, 1 in Korea, and 1 in Taiwan are involved in the study . Failure was defined as implant removal for any reason . The influence of mobility at placement, implant length and diameter, incision type, augmentation, crestal bone reduction, bone density, and the use of the operating room or dental clinic on survival were evaluated over 18 months . Crestal bone loss between placement and uncovering was also determined . RESULTS: Crestal bone loss ranged from 0.2 to 0.5 mm . The overall success rate from placement to 18 months was 96.6% . Implants mobile at placement failed more frequently (16.9%) compared with stable implants (3.1%) . Wide-diameter implants and longer implants exhibited higher survival rates . Incision design and surgery location did not influence survival . Bone density was important to clinical survival.

Ann Agric Environ Med, 2002, 9(2), 183 - 97
Toxin producing micromycetes on fruit, berries, and vegetables; Lugauskas A et al.; In 1999-2001 the investigations on mycological state of stored and sold fruit, berries, and vegetables grown in Lithuania and imported from other countries were performed . The samples of foodstuff were taken from storehouses, various supermarkets, and market places . Such ecological conditions lead to a rapid spreading of micromycetes and contamination of other articles of food stored and sold nearby . On fresh fruit and berries the development of microorganisms is slow . However, microorganisms penetrate into internal tissues of berries and fruit, thus becoming difficult to notice visually . Some microorganisms, especially micromycetes of some species belonging to the Penicillium Link, Aspergillus Mich . ex Fr., and other genera, are able to produce secondary metabolites (mycotoxins) of various compositions that are toxic to plants, animals, and humans . Therefore, the ability of micromycetes to synthesise and excrete toxic secondary metabolites was examined . Considering this issue, 393 micromycete strains ascribed to 54 genera and 176 species were tested . 46 strains were identified as active producers of toxic substances and were selected for further examinations . Most of them belonged to the Penicillium, Aspergillus and Fusarium genera . Their detection frequency on the investigated berries, fruit, and vegetables was determined, and the impact upon warm-blooded animals (BALB/c mice) was tested . Significant changes of the internal organs and blood composition were found in mice infected with toxic micromycetes . In conclusion, it was evidenced that more than 10% of micromycete strains developing on incorrectly-preserved fruit, berries and vegetables, produce toxic secondary metabolites that pose a potential health hazard for people eating or handling the foodstuff.

Int Microbiol, 2002 Dec, 5(4), 215 - 22 Epub 2002 Aug 27.
New approaches to the study of Antarctic lithobiontic microorganisms and their inorganic traces, and their application in the detection of life in Martian rocks; Ascaso C et al.; Microbial life in the harsh conditions of Antarctica's cold desert may be considered an analogue of potential life on early Mars . In order to explore the development and survival of this epilithic and endolithic form of microbial life, our most sophisticated, state-of-the-art visualization technologies have to be used to their full potential . The study of any ecosystem requires a knowledge of its components and the processes that take place within it . If we are to understand the structure and function of each component of the microecosystems that inhabit lithic substrates, we need to be able to quantify and identify the microorganisms present in each lithobiontic ecological niche and to accurately characterize the mineralogical features of these hidden microhabitats . Once we have established the techniques that will allow us to observe and identify these microorganisms and mineral substrates in situ, and have confirmed the presence of water, the following questions can be addressed: How are the microorganisms organized in the fissures or cavities? Which microorganisms are present and how many are there? Additional questions that logically follow include: What are the existing water relationships in the microhabitat and what effects do the microorganisms have on the mineral composition? Mechanical and chemical changes in minerals and mineralization of microbial cells can give rise to physical and/or chemical traces (biomarkers) and to microbial fossil formation . In this report, we describe the detection of chains of magnetite within the Martian meteorite ALH84001, as an example of the potential use of SEM-BSE in the search for plausible traces of life on early Mars.

Int Microbiol, 2002 Dec, 5(4), 177 - 88 Epub 2002 Nov 07.
Microbial mats and the search for minimal ecosystems; Guerrero R et al.; This article reviews some ecological concepts common to all kinds of ecosystems, describes the characteristics of microbial mats, and focuses on the description of the Ebro Delta microbial mats, to assess whether they fit the concept of a minimal ecosystem . First, microorganisms as components of ecosystems are considered, and some features of microbial life, including ubiquity, size and metabolism, genetic versatility, and strategies to overcome unfavorable conditions, are discussed . Models for ecosystems, regardless of their size, have the same basic components; tropical forests, multilayered planktonic microbial communities, and benthic microbial mats are analogous ecosystems at different scales . The structure--in terms of populations and communities--and ecophysiology of microbial mats are also discussed . The linear distribution of microbial populations along steep gradients of light and hydrogen sulfide allows for the simultaneous presence of different microbial populations . Defining the minimal ecosystem requirements necessary for the survival and proliferation of organisms is crucial in the search for extraterrestrial life and for establishing ecosystems beyond the Earth.

Infect Immun, 2003 Jan, 71(1), 271 - 4
Interleukin-1 receptor antagonist gene polymorphism, vaginal interleukin-1 receptor antagonist concentrations, and vaginal ureaplasma urealyticum colonization in pregnant women; Barton PT et al.; Ureaplasma urealyticum is the microorganism most frequently isolated from amniotic fluids of women in preterm labor . The relationship between vaginal colonization with U . urealyticum, vaginal interleukin-1 receptor antagonist (IL-1ra) levels, and the IL-1ra genotype in pregnant women was examined . Vaginal specimens, obtained with a cotton swab from 207 women in their first trimester of pregnancy, were tested for IL-1ra concentrations by enzyme-linked immunosorbent assay and for U . urealyticum and IL-1ra genotypes by PCR . U . urealyticum was detected in 85 (41.1%) women . The median IL-1ra level was 450 ng/ml in women positive for U . urealyticum, as opposed to 225 ng/ml in women negative for this microorganism (P < 0.0001) . Sixty-two percent of the 16 women who were homozygous for allele 2 of the IL-1ra gene (IL-1RN*2) were colonized with U . urealyticum, as opposed to 47% of the 49 women who were IL-1RN*1/IL-1RN*2 heterozygotes and 34% of the 133 women who were IL-1RN*1 homozygotes (P < 0.05) . Median IL-1ra levels were 750 ng/ml in IL-1RN*2 homozygotes, 300 ng/ml in IL-1RN*1/IL-1RN*2 heterozygotes, and 250 ng/ml in IL-1RN*1 homozygotes (P = 0.02) . The vast majority of subjects had an uneventful pregnancy and delivered a healthy infant at term . The IL-1ra genotype or U . urealyticum colonization was unrelated to birth weight . Pregnant women who are colonized with U . urealyticum during the first trimester have elevated vaginal IL-1ra concentrations and a higher prevalence of the IL-1RN*2 homozygote genotype than do noncolonized women.

Infect Immun, 2003 Jan, 71(1), 61 - 7
Interaction of pulmonary surfactant protein C with CD14 and lipopolysaccharide; Augusto LA et al.; In addition to their effects on alveolar surface tension, some components of lung surfactant also have immunological functions . We found recently that the hydrophobic lung surfactant protein SP-C specifically binds to the lipid A region of lipopolysaccharide (LPS) . In this study, we show that SP-C also interacts with CD14 . Four observations showed cross talk between the three molecules SP-C, LPS, and CD14 . (i) Like LBP, SP-C allows the binding of a fluorescent LPS to cells expressing CD14 (the other surfactant components were ineffective) . (ii) Recombinant radiolabeled CD14 and SP-C (or a synthetic analog of SP-C) interact in a dose-dependent manner . (iii) LPS blocks the binding of radiolabeled CD14 to SP-C-coated wells . (iv) SP-C enhances the binding of radiolabeled CD14 to LPS-coated wells . These results, obtained with native murine SP-C and with three synthetic analogs, suggest that LPS and CD14 interact with the same region of SP-C and that binding of SP-C modifies the conformation of CD14 or the accessibility of its LPS-binding site, allowing it to bind LPS . This ability of SP-C to interact with the pattern recognition molecule CD14 extends the possible immunological targets of SP-C to a large panel of microorganisms that can enter the airways.

Infect Immun, 2003 Jan, 71(1), 22 - 9
Resistance of macrophages to Mycobacterium avium is induced by alpha2-adrenergic stimulation; Weatherby KE et al.; The ability of macrophages to control the growth of microorganisms is increased by macrophage activation . Previously, it was shown that epinephrine activated mouse macrophages to resist the growth of Mycobacterium avium via alpha(2)-adrenergic stimulation . In the present study, we show that the alpha(2)-adrenergic agonist (alpha(2)-agonist) clonidine induced resistance to M . avium growth in the RAW264.7 mouse macrophage cell line . The ability of catecholamines to induce resistance to mycobacteria was specific to alpha(2)-adrenergic stimulation, as alpha(1)-, beta(1)-, and beta(2)-agonists had no effect . Receptor signaling through Gi proteins was required . A G-protein antagonist specific for the alpha subunits of the Go/Gi family blocked the increased resistance induced by clonidine, while a Gs-protein antagonist was without effect . Both nitric oxide (NO) production and superoxide (O(2)(-)) production were required for the increased resistance to M . avium growth induced by clonidine . Although NO production was required, clonidine did not increase the level of NO in M . avium-infected cells . Since NO and O(2)(-) interact to produce peroxynitrite (ONOO(-)), we examined whether ONOO(-) mediates the increased resistance to M . avium induced by clonidine . 5,10,15,20-Tetrakis(4-sulfonatophenyl)prophyrinato iron (III) chloride (FeTPPS), a specific scavenger of ONOO(-), inhibited the effect of clonidine on M . avium growth . Clonidine also increased the production of ONOO(-) in M . avium-infected RAW264.7 cells, as measured by the oxidation of 123-dihydrorhodamine and the production of nitrated tyrosine residues . We therefore conclude that alpha(2)-adrenergic stimulation activates macrophages to resist the growth of M . avium by enhancing the production of ONOO(-).

Immunol Rev, 2002 Dec, 190, 9 - 25
Single nucleotide polymorphisms in innate immunity genes: abundant variation and potential role in complex human disease; Lazarus R et al.; Under selective pressure from infectious microorganisms, multicellular organisms have evolved immunological defense mechanisms, broadly categorized as innate or adaptive . Recent insights into the complex mechanisms of human innate immunity suggest that genetic variability in genes encoding its components may play a role in the development of asthma and related diseases . As part of a systematic assessment of genetic variability in innate immunity genes, we have thus far have examined 16 genes by resequencing 93 unrelated subjects from three ethnic samples (European American, African American and Hispanic American) and a sample of European American asthmatics . Approaches to discovering and understanding variation and the subsequent implementation of disease association studies are described and illustrated . Although highly conserved across a wide range of species, the innate immune genes we have sequenced demonstrate substantial interindividual variability predominantly in the form of single nucleotide polymorphisms (SNPs) . Genetic variation in these genes may play a role in determining susceptibility to a range of common, chronic human diseases which have an inflammatory component . Differences in population history have produced distinctive patterns of SNP allele frequencies, linkage disequilibrium and haplotypes when ethnic groups are compared . These and other factors must be taken into account in the design and analysis of disease association studies.

J Clin Periodontol, 2002 Dec, 29(12), 1065 - 71
Quantitative assessment of apoptotic and proliferative gingival keratinocytes in oral and sulcular epithelium in patients with gingivitis and periodontitis; Jarnbring F et al.; BACKGROUND: Periodontal disease is caused by a chronic infection inducing an inflammatory reaction that leads to a breakdown of tooth-supporting tissue . The maintenance of an equilibrium between the host defence and microorganisms in the sulcus is essential to preserve health . All multicellular organisms have mechanisms for killing their own cells, and use physiological cell death for defence, development, homeostasis and ageing . Apoptosis and proliferation are very important phenomena in regulating this and a disturbance is often associated with disease e.g . cancer, AIDS, Alzheimer's disease, rheumatoid arthritis . OBJECTIVE: The aim of this study was to determine whether the number of apoptotic and proliferative gingival keratinocytes differed between patients with gingivitis and those with periodontitis . MATERIAL AND METHODS: The distribution of neutrophil elastase, PCNA/cyclin, DNA fragmentation (apoptosis) and p53 was determined with immunocytochemical techniques . We used paraffin-embedded sections from gingival biopsies and did quantitative analyses . RESULTS AND CONCLUSION: These showed that 5-12% of the keratinocytes in the basal layers of the epithelium proliferated in the two groups . Fewer apoptotic cells were seen in the oral epithelium than in the sulcus in all subjects in both groups . Only in the most apical part of the sulcus, close to the junctional epithelium, did the number of apoptotic keratinocytes exceed the proliferative ones in patients with periodontitis.

Semin Pediatr Infect Dis, 2002 Oct, 13(4), 249 - 56
Ehrlichiosis in children; Lantos P et al.; Ehrlichia are obligate intracellular bacteria that belong to the family Rickettsiaceae . Human monocytic ehrlichiosis (HME) and human granulocytic ehrlichiosis (HGE) are the 2 ehrlichial diseases that are of greatest health concern in the United States . The agents causing HME and HGE are zoonotic pathogens requiring a mammalian reservoir and an arthropod vector . Differences in the geographic distribution of their tick vectors account for the concentration of HME in the South and southeastern United States and HGE in the Northeast and northern Midwest . Both infections have been reported in coastal regions from Rhode Island to Florida . HME and HGE are flulike illnesses that usually are self-limited but may be fatal . Diagnosis is confirmed by identification of the microorganism on blood smear or polymerase chain reaction or by detection of anti-ehrlichial antibody . Doxycycline is the antibiotic of choice for treatment of ehrlichiosis .

Isr Med Assoc J, 2002 Nov, 4(11), 1023 - 4
The leukocyte NADPH oxidase; Babior BM; The leukocyte NADPH oxidase catalyzes the reduction of oxygen to O2- (superoxide) at the expense of NADPH . The O2- then dismutes to H2O2, which serves to oxidize Cl- to HOCl, a potent microbicidal agent that is used by leukocytes to kill invading microorganisms . This oxidation is catalyzed by myeloperoxidase . O2 is also used to make other microbicidal oxidants, some in reactions with nitric oxide . The oxidase itself is highly complex, consisting of four unique subunits and Rac2 . In the resting cell, two of the subunits, p22PHOX and gp91PHOX, are located in the membrane, and the other two, p47PHOX and p67PHOX, are in the cytosol . The electron-carrying components of the oxidase are located in gp91PHOX; the NADPH binding site is generally regarded to be in gp91PHOX as well, but there is some evidence that it may be in p67PHOX . When the oxidase is activated, p47PHOX is phosphorylated at specific sites, and the cytosolic components plus Rac2 migrate to the membrane to assemble the active oxidase.

Cad Saude Publica, 2002 Nov-Dec, 18(6), 1665 - 74
{From miasmas to microorganisms: hygienic lower-class housing}; Caponi S; By analyzing the influence of early Pasteurian thought on health discourse and practice, this paper discusses the thesis postulating an opposition between pre- and post- Pasteurian hygiene . Focusing on an analysis of the sanitary strategies to control lower-class housing that were proposed by Argentine and Brazilian hygienists, the author identifies the breaks and continuities between classical hygiene and hygiene reorganized by microbiology.

J Leukoc Biol, 2002 Dec, 72(6), 1180 - 9
CD40 ligation and phagocytosis differently affect the differentiation of monocytes into dendritic cells; Rosenzwajg M et al.; That monocytes can differentiate into macrophages or dendritic cells (DCs) makes them an essential link between innate and adaptive immunity . However, little is known about how interactions with pathogens or T cells influence monocyte engagement toward DCs . We approached this point in cultures where granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-4 induced monocytes to differentiate into immature DCs . Activating monocytes with soluble CD40 ligand (CD40L) led to accelerated differentiation toward mature CD83(+) DCs with up-regulated human leukocyte antigen-DR, costimulatory molecules and CD116 (GM-CSF receptor), and down-regulation of molecules involved in antigen capture . Monocytes primed by phagocytosis of antibody-opsonized, killed Escherichia coli differentiated into DCs with an immature phenotype, whereas Zymosan priming yielded active DCs with an intermediate phenotype . Accordingly, DCs obtained from cultures with CD40L or after Zymosan priming had a decreased capacity to endocytose dextran, but only DCs cultured with CD40L had increased capacity to stimulate allogeneic T cells . DCs obtained after E . coli or Zymosan priming of monocytes produced high levels of proinflammatory tumor necrosis factor alpha and IL-6 as well as of regulatory IL-10, but they produced IL-12p70 only after secondary CD40 ligation . Thus, CD40 ligation on monocytes accelerates the maturation of DCs in the presence of GM-CSF/IL-4, whereas phagocytosis of different microorganisms does not alter and even facilitates their potential to differentiate into immature or active DCs, the maturation of which can be completed upon CD40 ligation . In vivo, such differences may correspond to DCs with different trafficking and T helper cell-stimulating capacities that could differently affect induction of adaptive immune responses to infections.

AIDS Res Hum Retroviruses, 2002 Nov 20, 18(17), 1311 - 7
High mannose glycans and sialic acid on gp120 regulate binding of mannose-binding lectin (MBL) to HIV type 1; Hart ML et al.; Mannose-binding lectin (MBL) is a C-type lectin of the innate immune system that binds to carbohydrates on the surface of certain microorganisms . Previous studies showed that MBL binds to gp120, the envelope glycoprotein of HIV-1 . gp120 is extensively glycosylated, with N-linked complex and high mannose carbohydrates accounting for about half of the molecular weight . The objectives of this study were to determine the types of glycans on gp120 important for MBL binding and to determine if alteration of complex glycans with neuraminidase (NA) could enhance the interaction of MBL with virus . Lectin blot analyses revealed that MBL interacted with recombinant gp120 (rgp120) from both T cell-tropic and M-tropic virus strains . Treatment of rgp120 with endoglycosidase H (eH) or endoglycosidase F1 (eF1) abrogated binding of MBL, but did not decrease binding of wheat germ agglutinin indicating that high mannose and/or hybrid N-linked glycans were required for MBL binding . Removal of sialic acids from rgp120 with NA enhanced MBL binding . Treatment of intact virus from T cell lines or primary isolates with eF1 also significantly decreased HIV binding to MBL, while treatment with NA substantially increased binding . Treatment of virus with both eF1 and NA did not decrease binding compared to NA alone suggesting that NA treatment exposed binding sites on gp120 that are not high mannose glycans . These studies provide evidence that MBL binds to HIV via high mannose carbohydrates on gp120 and shows that the interaction of MBL with virus is regulated by sialylation.

Crit Rev Food Sci Nutr, 2002, 42(6), 627 - 45
Food processing by high hydrostatic pressure; San Martin MF et al.; The use of high hydrostatic pressures (HHP) for food processing is finding increased application within the food industry . One of the advantages of this technology is that because it does not use heat, sensory, and nutritional attributes of the product remain virtually unaffected, thus yielding products with better quality than those processed traditional methods . HHP have the ability to inactivate microorganisms as well as enzymes responsible for shortening the life of a product . In addition to lengthening the shelf-life of food products, HHP can modify functional properties of components such as proteins, which in turn can lead to the development of new products . Equipment for large-scale production of HHP processed products are commercially available nowadays . Guacamole, sliced ham, oysters, and fruit juices are some of the products currently available on the market . HHP technology is one of the most promising nonthermal processes.

Transpl Infect Dis, 2002, 4 Suppl 3, 46 - 51
Prevention of fungal infection in transplantation; Paya CV; Despite significant advances in the diagnosis, prevention, and treatment of fungal infection in transplant recipients, this infection complication remains a major cause of morbidity and mortality . Our understanding of the pathogenesis of the different fungal microorganisms has enabled us to identify patients at risk for such infections . While Candida infection remains a major complication in patients with intra-abdominal solid organ transplantations in which the bowel is surgically manipulated, Aspergillus infection remains the main fungal complication in lung transplantation recipients . The incidence of all types of fungal infection remains around 5-10%, while mortality following Aspergillus infection remains around 70% . Suppression of Candida growth at the time of surgical manipulation of the bowel should be the mainstay of prevention of this infection in intra-abdominal organ transplantation . Fluconazole is effective and relatively safe at 100-400 mg daily for the first 1-3 months post-liver transplantation . Prevention strategies toward Aspergillus infections remain elusive, but a number of manipulations, such as inhaled liposomal preparations post-organ transplantation or the preemptive use or universal prophylaxis of itraconazole are being validated . The next step is to determine the clinical value of molecular diagnostic techniques for the identification and preemptive therapy of patients at risk for the variety fungal infections.

MIC, Bioengineering, Cell Biology, Microbe, Pcr, Microorganisms, Bioremediation, Microorganism, Amino Acid, Microbiology, Bioreactor, Bacteria, Water Purification, Microbes, d, Dna, g, Bacteriology, p, Molecular Microbiology, t, Bacterium, e, Activated Sludge, Genetic Engineering, Antibiotic resistance, s, Thermophilic, b, Haemophilus, e, Escherichia coli, m, Antibacterial, f, Bacillus subtilis, s, Yeasts, s, Escherichia coli, b, Agrobacteria, s, Multidrug resistant, b, E. coli, p, Listeriosis, c, Antibacterial, y, Biodegradation, e, Pseudomonas, c, Bacterium, e, P. aeruginosa, s, Staphylococci, g, Phages, m, Bacterial, s, Cell culture, b, Bacillus, c, Acinetobacter, s, Streptococcal, s, Yeasts, s, Antibiotics, p, Bacteroides, b, Petri dish, s, Pseudomonas, Cholera,

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

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

Last modified: May 25, 2005