Phytochemistry, 2003 Aug, 63(8), 959 - 64 Glucosides from Vitex agnus-castus; Kuruuzum-Uz A et al.; The methanolic extract of the flowering stems of Vitex agnus-castus yielded three new iridoids: 6'-O-foliamenthoylmussaenosidic acid (agnucastoside A), 6'-O-(6,7-dihydrofoliamenthoyl)mussaenosidic acid (agnucastoside B) and 7-O-trans-p-coumaroyl-6'-O-trans-caffeoyl-8-epiloganic acid (agnucastoside C) in addition to four known iridoids (aucubin, agnuside, mussaenosidic acid and 6'-O-p-hydroxybenzoylmussaenosidic acid) and one known phenylbutanone glucoside (myzodendrone) . The structure elucidations were mainly done by spectroscopic methods (1D and 2D NMR spectra) and MS data interpretation . The purified compounds were tested for biological activities against various microorganisms and cancer cell lines. Nitric Oxide, 2003 Jun, 8(4), 222 - 30 Cell density-enhanced expression of inducible nitric oxide synthase in murine macrophages mediated by interferon-beta; Jacobs AT et al.; Nitric oxide (NO) has an important cytotoxic role in host defense processes against invading microorganisms and neoplastic cells . Here we demonstrate the effect of culture density on the expression of NO synthase and NO production by lipopolysaccharide (LPS)-activated RAW 264.7 macrophages . At high cell densities, the LPS-induced expression of iNOS message, protein, and activity is markedly enhanced . We demonstrate the effects to be mediated by a diffusible macrophage product . Increasing cell density correlates with activation of IFN-dependent signaling pathways . We observe enhanced phosphorylation of STAT-1 on tyrosine 701 and serine 727, and an increase in STAT-1 DNA binding . Expression of the IFN-stimulated transcription factor IRF-1 is also enhanced . The data are consistent with the reported involvement of IFN-beta as an autocrine co-activator of iNOS expression . Considering the importance of NO as a cytotoxic mediator of host immunity, the data suggest that macrophage density is important in regulating the magnitude of NO production, and thus, the host response to infection. Bioelectrochemistry, 2003 Aug, 60(1-2), 29 - 36 Electrochemical study of heavy metals and metallothionein in yeast Yarrowia lipolytica; Strouhal M et al.; The bioaccumulation of heavy metals (cadmium, nickel, cobalt and zinc) and the effect of these metals on the production of metallothionein and metallothionein-like proteins (MT) in Yarrowia lipolytica was studied by electrochemical methods . The concentrations of heavy metals were determined by differential pulse voltammetry (DPV) . A combination of the constant current chronopotentiometric stripping analysis (CPSA) and adsorptive transfer stripping technique (AdTS) was used to determine the content of MT in cells . Both the bioaccumulation of heavy metals and the production of MT in different cell compartments of Y . lipolytica exposed to heavy metals were monitored . The LD(50) of each metal was determined from the number of viable cells in yeast cultures: LD(50)Cd (37.5 microM), LD(50)Ni (570 microM), LD(50)Co (700 microM), and LD(50)Zn (1800 microM) . The highest concentrations of heavy metals were found in the cell wall and membrane debris while the lowest concentrations were detected in the cytoplasm . Cadmium and nickel showed the most significant effect on the production of MT . This study provides new insights into the ecophysiology of microorganisms and demonstrates the potential use of these electrochemical methods in biotechnology. Biotechnol Lett, 2003 Jun, 25(11), 883 - 6 A peptide antibody for rapid screening of Streptomyces species producing phospholipase D; Uhm TB et al.; By examining the conserved regions in the protein sequences of eight different Streptomyces phospholipase Ds (PLD) reported so far and the X-ray crystallographic structure of a Streptomyces PLD, we designed a peptide sequence, DPANRGAVGSGGYSQIKSL, for the screening of microorganisms producing PLD . In the enzyme-linked immunosorbent assay using a mouse antibody raised against the designed peptide, we recovered seven producing strains out of 128 soil isolates. J Nutr, 2003 Aug, 133(8), 2635 - 8 Iron-deficient mice fail to develop autoimmune encephalomyelitis; Grant SM et al.; Determinations of the effects of iron status on the immune system are complicated by the fact that microorganisms and immune cells both utilize iron . To determine the role of iron in immune function, we utilized a model {experimental autoimmune encephalomyelitis (EAE)} in which a strong antigen-specific CD4+ T-cell response develops in the absence of infection . EAE is an autoimmune disease frequently used as a model for the human disease multiple sclerosis (MS) . EAE was induced in B10.PL mice fed low iron (1 mg/kg), normal iron (10 mg/kg) or high iron (160 mg/kg) diets that were replete in all other nutrients . Liver iron measurements verified iron status, i.e., low iron mice had 1.9 micro mol/g tissue, normal iron mice, 3.27 micro mol/g tissue and high iron mice, 5.35 micro mol/g tissue . EAE symptoms were most severe in normal iron mice, and EAE did not develop in low iron mice . The incidence of EAE was 71% in normal iron mice, 62% in iron-overloaded mice and 0% in iron-deficient mice . Two of seven mice in the normal iron group developed severe EAE and were euthanized . None of the iron-overloaded mice developed severe EAE . Other measures of EAE severity were similar in the normal and iron-overloaded mice . The data suggest that iron deficiency provides protection from the development of EAE and that iron excess with its potential contribution to free radical formation was not an important factor . The mechanism of EAE inhibition in iron-deficient mice likely involves the delivery and metabolism of iron for optimal CD4+ T-cell development. Zhonghua Kou Qiang Yi Xue Za Zhi, 2003 May, 38(3), 206 - 9 {Effect of spirochetes and porphyromonus gingivalis on oral malodor}; Zhu WD et al.; OBJECTIVE: To assess the effect of Spirochetes and Porphyromonus gingivalis (Pg) on the production of volatile sulfur compounds (VSCs) and oral malodor, respectively . METHODS: 117 systemically healthy periodontitis patients were divided into two groups according to organoleptic rating (OR) by organoleptic method: 85 subjects with oral malodor (halitosis group, HG) and 32 cases without oral malodor (no-halitosis group, NHG) . VSCs measurements were made with halimeter . Subgingival plaques and tongue coating were taken to calculate the proportion of Spirochetes (with 2% Congo Red Smears, S%) and the amounts of Pg (with indirect immunofluorescence method) . The examination will be repeated in 46 HG cases after periodontal therapy . RESULTS: S% in PD < 4 mm sites of HG was significantly higher than that in NHG (P < 0.01) . S% in PD >or= 4 mm sites and tongue coating, Pg in all three sites were not different between two groups . In HG, OR and VSCs levels were correlated with S% in tongue coating (P < 0.01, P < 0.05) and PD < 4 mm sites (P < 0.05), and not correlated with S% in PD >or= 4 mm sites . OR was related to Pg in PD >or= 4 mm sites (P < 0.05) and there was no relationship between OR and Pg in tongue coating and PD < 4 mm sites . VSCs levels was not correlated with Pg in all three sites . S% in tongue coating and subgingival plaque, Pg in subgingival plaque, OR and VSCs levels were significantly reduced after periodontal therapy (P < 0.01) . CONCLUSIONS: The direct relationship of Spirochetes and Pg respectively to bad breath was not found obviously . But Spirochetes is one of the microorganism on oral malodor . Periodontal therapy can improve oral malodor. Braz J Med Biol Res, 2003 Aug, 36(8), 987 - 91 Epub 2003 Jul 23. Endothelial cells, tissue factor and infectious diseases; Lopes-Bezerra LM et al.; Tissue factor is a transmembrane procoagulant glycoprotein and a member of the cytokine receptor superfamily . It activates the extrinsic coagulation pathway, and induces the formation of a fibrin clot . Tissue factor is important for both normal homeostasis and the development of many thrombotic diseases . A wide variety of cells are able to synthesize and express tissue factor, including monocytes, granulocytes, platelets and endothelial cells . Tissue factor expression can be induced by cell surface components of pathogenic microorganisms, proinflammatory cytokines and membrane microparticles released from activated host cells . Tissue factor plays an important role in initiating thrombosis associated with inflammation during infection, sepsis, and organ transplant rejection . Recent findings suggest that tissue factor can also function as a receptor and thus may be important in cell signaling . The present minireview will focus on the role of tissue factor in the pathogenesis of septic shock, infectious endocarditis and invasive aspergillosis, as determined by both in vivo and in vitro models. Nippon Geka Gakkai Zasshi, 2003 Jul, 104(7), 518 - 22 {Molecular diagnosis of infectious disease}; Fukushima R et al.; With the development of molecular biology over the decades, molecular technologies have become available for clinical diagnosis of infectious disease . The principle of the technique is to detect specific genes of microorganisms . Compared with the traditional culturing techniques, molecular technologies have several advantages: 1) direct detection and rapid identification of organisms that are slow growing or those currently lacking a system for in vitro cultivation; 2) nucleic acid-based methods for epidemiologic typing of microorganisms; and 3) identification of genotypic markers of microbial resistance to specific antibiotics . When sufficient amounts of nucleic acid can be obtained, direct probe tests (simple molecular hybridization-based techniques) may be performed . In many instances with small amounts of sample nucleic acids, amplification techniques(e.g., polymerase chain reaction{PCR}) are extremely useful . However, the application of PCR to clinical specimens has potential pitfalls due to the susceptibility of PCR to inhibitors, contamination, and experimental conditions . In some cases, DNA fingerprinting or restriction fragment-length polymorphism analysis is used and it has become the method of choice for epidemiologic analysis . The major disadvantage of the molecular method is that we cannot differentiate whether the microorganisms identified are alive or dead and chemosensitivity testing cannot be performed. Semin Pediatr Infect Dis, 2003 Apr, 14(2), 90 - 5 Acute disseminated encephalomyelitis; Dale RC; Acute disseminated encephalomyelitis (ADEM) is a monophasic inflammatory disorder of the central nervous system (CNS) . Unlike viral encephalitis, microorganisms do not invade the CNS . Instead, ADEM is a postinfectious disease mediated by auto-reactive cells or molecules . Clinical characteristics of ADEM are consistent with disseminated involvement of the CNS, including encephalopathy and pyramidal, cerebellar, and brainstem signs . Bilateral optic neuritis and transverse myelitis are particularly suggestive of demyelinating diseases such as ADEM . Unlike viral encephalitis, seizures rarely are a prominent symptom . The most useful diagnostic investigation is magnetic resonance neuroimaging that commonly shows multifocal lesions throughout the brain and spinal cord . As ADEM is an immune-mediated disorder, treatment includes immunomodulatory therapies (particularly steroids), although no clinical trials have been performed to define the most efficacious agent . In view of the treatment differences between ADEM and viral encephalitis, being familiar with ADEM is essential for pediatricians managing acute neurological disorders . Science, 2003 Aug 15, 301(5635), 976 - 8 Epub 2003 Jul 24. Geographic barriers isolate endemic populations of hyperthermophilic archaea; Whitaker RJ et al.; Barriers to dispersal between populations allow them to diverge through local adaptation or random genetic drift . High-resolution multilocus sequence analysis revealed that, on a global scale, populations of hyperthermophilic microorganisms are isolated from one another by geographic barriers and have diverged over the course of their recent evolutionary history . The identification of a biogeographic pattern in the archaeon Sulfolobus challenges the current model of microbial biodiversity in which unrestricted dispersal constrains the development of global species richness. Protein Expr Purif, 2003 Aug, 30(2), 210 - 8 Expression and purification of Arg196 and Lys272 mutants of mevalonate kinase from Methanococcus jannaschii; Chu X et al.; In microorganisms and plants, mevalonate kinase is involved in the biosynthesis of isoprenoid derivatives, one of the largest groups of natural products . We subcloned the gene of mevalonate kinase from Methanococcus jannaschii into a bacterial expression vector pLM1 with six continuous histidine codons attached to the 5' end of the gene . A variety of mutant expression plasmids including pMMK(R196K), pMMK(R196Q), pMMK(R196V), pMMK(K272R), and pMMK(K272A) have been constructed using site-directed mutagenesis . The wild-type protein and mutants were overexpressed and purified with a nickel HiTrap chelating metal affinity column to homogeneity . CD spectroscopy of wild-type protein and mutants indicates that none of the above mutations induces significant secondary structural changes . The results from kinetic studies showed that Arg196 is an essential residue for the function of the enzyme . Kinetic studies of Lys272 mutants indicate that salt bridge Lys272-Glu14 plays an important role in maintaining the active site microenvironment that is essential for catalytic activity of the enzyme. J Oral Rehabil, 2003 Aug, 30(8), 790 - 5 Confocal laser scanning microscopic analysis of early plaque formed on resin composite and human enamel; Konishi N et al.; The purpose of this study was to analyse quantitatively the early bacterial plaque formed on resin composite and human enamel in vivo, using a confocal laser scanning microscope . Test pieces of resin composite and human enamel were retained at the buccal surfaces of the upper first molars of three volunteers for 4, 8 and 24 h to allow plaque formation . Then, the specimens were immersed in propidium iodide in phosphate-buffered saline to stain adherent bacteria and observed with a confocal laser scanning microscope . The ratios of the area occupied by microorganisms to the whole area of the optical field were calculated using a photo-image analysis system . The thickness of the plaque was also measured . Quantitative analysis revealed that the resin composite showed significantly higher bacterial adherence than human enamel throughout the test period . A difference was noticed in the morphology of the bacteria between the two groups . Our findings suggest that resin composite shows higher bacteria adherence during early plaque formation compared with human enamel . In addition, the present findings may suggest a presence of the difference in bacterial composition of plaque in both specimens. Ceska Gynekol, 2003 May, 68(3), 206 - 9 {Specific mucosal immunity in the female reproductive tract: general introduction (part 1)}; Zivny J et al.; Immune mediated defense against pathogenic microorganisms and maintenance of immune tolerance to allogeneic antigens of sperm and fetus are main function of female reproductive tract mucosal immune system . Basic mechanisms of mucosal immune system and the unique features of female reproductive tract immune system are reviewed in this article. Appl Microbiol Biotechnol, 2004 Jan, 63(4), 445 - 51 Epub 2003 Jul 15. Inorganic tin and organotin interactions with Candida maltosa; White JS et al.; As a consequence of the widespread industrial and agricultural applications of organotins, contamination of various ecosystems has occurred in recent decades . Understanding how these compounds interact with microorganisms is important in assessing the risks of organotin pollution . The organotins, tributyltin (TBT), trimethyltin (TMT) and inorganic tin, Sn(IV), were investigated for their physical interactions with non-metabolising cells and protoplasts of the yeast Candida maltosa, an organism that is often associated with contaminated environments . Uptake, toxicity and membrane-acting effects of these compounds, at concentrations approximating those found in polluted environments, were assessed . Sn(IV) and TBT uptake occurred by different mechanisms . Uptake of Sn(IV) was 2-fold greater in intact cells than protoplasts, underlining the importance of cell wall binding, whereas TBT uptake levels by both cell types were similar . TBT uptake resulted in cell death and extensive K+ leakage, while Sn(IV) uptake had no effect . TMT did not interact with cells . Of the three compounds, TBT alone altered membrane fluidity, as measured by the fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene incorporated into cells . Anisotropy of 1-(4-trimethylaminophenyl-6-phenyl-1,3,5-hexatriene) was not affected, implying that TBT is not confined to the surface of the cytoplasmic membrane, but acts within membrane lipids . These results indicate that the cell wall is the dominant site of Sn(IV) interactions with yeast, while lipophilic interactions play an important role in uptake and toxicity of TBT. Appl Microbiol Biotechnol, 2003 Oct, 62(5-6), 517 - 22 Epub 2003 Jul 16. Vesicular transport route of horseradish C1a peroxidase is regulated by N- and C-terminal propeptides in tobacco cells; Matsui T et al.; Peroxidases (PRX, EC 1.11.1.7) are widely distributed across microorganisms, plants, and animals; and, in plants, they have been implicated in a variety of secondary metabolic reactions . In particular, horseradish (Armoracia rusticana) root represents the main source of commercial PRX production . The prxC1a gene, which encodes horseradish PRX (HRP) C, is expressed mainly in the roots and stems of the horseradish plant . HRP C1a protein is shown to be synthesized as a preprotein with both a N-terminal (NTPP) and a C-terminal propeptide (CTPP) . These propeptides, which might be responsible for intracellular localization or secretion, are removed before or concomitant with production of the mature protein . We investigated the functional role of HRP C1a NTPP and CTPP in the determination of the vesicular transport route, using an analytical system of transgenically cultured tobacco cells (Nicotiana tabacum, BY2) . Here, we report that NTPP and CTPP are necessary and sufficient for accurate localization of mature HRP C1a protein to vacuoles of the vesicular transport system . We also demonstrate that HRP C1a derived from a preprotein lacking CTPP is shunted into the secretory pathway. Med Wieku Rozwoj, 2003 Apr-Jun, 7(2), 271 - 7 {Role of mycoplasma pneumoniae infection in aetiopathogenesis of juvenile idiopatic arthritis}; Postepski J et al.; Although more and more is known about chronic autoimmune diseases, attempts to establish one trigger factor have been unsuccessful . The role of endogenic factors is beyond doubt . But it is emphasized that environmental factors are necessary to cause the disease . Infections are taken under consideration as trigger mechanism in the development of autoimmune diseases including chronic arthritis . Both numerous viruses and bacteria are among the microorganism mentioned . We considered it sensible to conduct research on Mycoplasma pneumoniae infections in a group of patients with juvenile idiopathic arthritis (JIA) . MATERIALS: 19 patients diagnosed with JIA aged between 6-17 were investigated for Mycoplasma pneumoniae infection whose blood was examined for antibodies against Mycoplasma pneumoniae in class IgG and IgM . The control group comprised 20 children of similar age admitted to hospital with digestive tract complaints . Methods: Serologic tests were made in serum . Marking of antigens of class IgM and IgG were made by Elisa method using commercial kits produced by Scientific Point . Quantitative calculations of a level of antigens were done using appropriate standards, positive and negative serum of reference standard and calibration curve . RESULTS: In 11 patients positive reaction for Mycoplasma pneumoniae in class IgG was observed and only in 2 in class IgM with low titer . In the control group positive reaction was observed in 3 children (15%) . The fact that 58% of patients were infected (contact either with Mycoplasma pneumoniae) indicates that in the group of our patients with JIA, infections with these bacteria might have had a role in triggering the disease. Expert Opin Pharmacother, 2003 Aug, 4(8), 1345 - 54 Antibiotic prophylaxis strategies in the prevention of ventilator-associated pneumonia; Sirvent JM et al.; Ventilator-associated pneumonia (VAP) is defined as a nosocomial pneumonia occurring > 48 h after endotracheal intubation . VAP may occur very early after intubation and it is usually defined as early-onset pneumonia, which occurs during the first 4 days . The occurrence of VAP is associated with an increase in morbidity and mortality . The pathogenesis of VAP often results from aspiration of colonised secretions in injured patients and this colonisation of the upper airway acts as a main risk factor in the development of pneumonia . It has been hypothesised that the bacterial inoculum may be decreased through the administration of systemic antibiotic prophylaxis . Antibiotic prophylaxis strategies to prevent VAP can be administered over an extended period to cover all microorganisms using selective digestive decontamination regimens, or in a short-term course of no more than 24 h . Probably, the second strategy is the most useful in the prevention of VAP because it has a lower impact on the emergence of bacterial resistance . This manuscript aims to review current opinions regarding antibiotic prophylaxis strategies in the prevention of VAP. PDA J Pharm Sci Technol, 2003 May-Jun, 57(3), 160 - 85 Measuring the thermal resistance of microorganisms: selecting an appropriate test system, correcting for heat-transfer lags, and determining minimum heating times; Pflug IJ; Errors that occur in physical systems used to evaluate the heat resistance of microorganisms are discussed: namely, (a) not knowing the test heating-medium temperature accurately, (b) using heating times that are so short that the maximum temperature reached in the test unit is significantly below the test heating-medium temperature, and (c) ignoring significant heat-transfer lags, first in the heating and later in the cooling of the test units . Procedures and methods that can be used to minimize the effect of potential test-system errors on microbial resistance data are reported . Examples are included regarding the treatment of the different types of errors . Heating and cooling lag-correction values for several commonly-used testing systems, gleaned from the published literature and from the author's experience, are listed . A method is described and illustrated regarding how we may determine (in advance of carrying out an experiment to gather enumeration or survivor-curve data), the shortest heating time--highest temperature that should be used with a specific test-unit system and microbial DT -value. Cell Physiol Biochem, 2003, 13(3), 165 - 72 Involvement of phosphatidic acid in both degranulation and oxidative activity in fMet-Leu-Phe stimulated polymorphonuclear leukocytes; Vocks A et al.; BACKGROUND/AIM: Polymorphonuclear leukocytes (neutrophils) release a variety of toxic agents--proteins and reactive oxygen species (ROS)--that are used to inactivate foreign microorganisms in the non-specific immune response . This study was undertaken to compare intracellular signalling pathways that lead to the ROS production as well as degranulation of azurophilic granules of human fMet-Leu-Phe/cytochalasin B stimulated neutrophils . METHODS: Luminol-amplified chemiluminescence was used for monitoring the oxidative activity of human neutrophils in the presence of various inhibitors . The elastase activity was assessed in the neutrophil supernatant as a marker for degranulation of azurophilic granules . RESULTS: Tested inhibitors of enzymes of signalling cascades showed the same effect on the ROS production and on the activity of elastase released from neutrophils . The only difference was obtained with staurosporine: it inhibited the chemiluminescence response, but increased the elastase release . CONCLUSION: Early signalling pathways leading to the ROS production and the degranulation are ubiquitous in human neutrophils . They are branching most probably at the point of the phosphatidic acid production by phospholipase D . A protein kinase activated by this lipid second messenger might play a central regulatory role in human neutrophils . Int Dent J, 2003, 53 Suppl 3, 200 - 7 Update on general health risk of periodontal disease; Slots J; OBJECTIVES: To review the potential of periodontal infections to cause nonoral diseases . Therapeutic recommendations are provided to help patients and dental practitioners prevent systemic complications from periodontal infections . FINDINGS: Systemic diseases from oral bacteria are mostly caused by transient bacteraemias, which can occur spontaneously from dental foci of infection, from mastication, brushing, flossing or other daily manipulations, or from dental treatments . Examples of systemic infections that may involve oral microorganisms include infective endocarditis, aspiration pneumonia, HIV-related disseminated candidiasis and cancrum oris, septicaemia associated with cancer chemotherapy and radiotherapy, necrotising faciitis and various other life-threatening infections . Inflamed gingiva constitutes a significant reservoir for herpes viruses, which have the potential to cause serious systemic diseases in immunocompromised patients . Periodontal disease may also aggravate chronic insulin insensitivity and thus interfere with glycaemic control in diabetic patients . Controversy surrounds the involvement of periodontal infections in coronary heart disease . CONCLUSIONS: Cumulative evidence suggests that periodontal disease can be an important cause of morbidity and mortality of various systemic diseases, especially in individuals exhibiting compromised host defence . Maintaining a healthy dentition and periodontium by means of daily oral hygiene practice and regular professional care is the most effective way of preventing systemic diseases from oral infections. Cardiol Clin, 2003 May, 21(2), 147 - 58 Progress toward a global understanding of infective endocarditis . Lessons from the International Collaboration on Endocarditis; Cabell CH et al.; In the century and a quarter since William Osler delivered his famed Gulstonian lectures on endocarditis, continual advancements have been made in understanding and treating this disease . Here we have reviewed some key aspects of current knowledge in the areas of population epidemiology, host factors, microorganisms, and diagnosis . The advent of the ICE investigation provides the opportunity to further expand our understanding of IE by developing a very large, global database of IE patients whose clinical, echocardiographic, and microbiologic findings have been characterized with standard methodology . Further, ICE may serve as a rich source of material for investigators seeking to perform specific studies . Finally, the ICE infrastructure creates the opportunity for performing randomized trials to test therapeutic strategies . Although many obstacles remain to be overcome, ICE has created the opportunity for a quantum leap in our knowledge of IE over the next 25 years. Infect Immun, 2003 Aug, 71(8), 4375 - 81 Transfected Plasmodium knowlesi produces bioactive host gamma interferon: a new perspective for modulating immune responses to malaria parasites; Ozwara H et al.; Transgenic pathogenic microorganisms expressing host cytokines such as gamma interferon (IFN-gamma) have been shown to manipulate host-pathogen interaction, leading to immunomodulation and enhanced protection . Expression of host cytokines in malaria parasites offers the opportunity to investigate the potential of an immunomodulatory approach by generating immunopotentiated parasites . Using the primate malaria parasite Plasmodium knowlesi, we explored the conditions for expressing host cytokines in malaria parasites . P . knowlesi parasites transfected with DNA constructs for expressing rhesus monkey (Macaca mulatta) IFN-gamma under the control of the heterologous P . berghei apical membrane antigen 1 promoter, produced bioactive IFN-gamma in a developmentally regulated manner . IFN-gamma expression had no marked effect on in vitro parasite development . Bioactivity of the parasite-produced IFN-gamma was shown through inhibition of virus cytopathic effect and confirmed by using M . mulatta peripheral blood cells in vitro . These data indicate for the first time that it is feasible to generate malaria parasites expressing bioactive host immunomodulatory cytokines . Furthermore, cytokine-expressing malaria parasites offer the opportunity to analyze cytokine-mediated modulation of malaria during the blood and liver stages of the infection. Cell Signal, 2003 Oct, 15(10), 901 - 9 Roles played by Ras subfamily proteins in the cell and developmental biology of microorganisms; Weeks G et al.; The Ras subfamily proteins are monomeric GTPases that function as molecular switches in cellular signal transduction pathways . This review describes our current knowledge of the roles that these proteins play in the growth and differentiation of single celled microorganisms. Kansenshogaku Zasshi, 2003 Jun, 77(6), 443 - 50 {Study of a respiratory syncytial virus diagnostic test kit based on immunochromatography}; Saikusa M et al.; We carried out clinical and basic studies of the Directigen Lateral Flow RSV (Becton, Dickinson and Company, USA), a rapid test kit that detects respiratory syncytial virus (hereinafter referred to as "RSV") antigens based on immunochromatography . For the clinical study, 103 nasopharyngeal aspirates from patients with acute respiratory infections were used to evaluate the kit . Compared to the cell culture method, the Directigen Lateral Flow RSV showed a sensitivity of 100% (16/16) and a specificity of 94.3% (82/87), and an agreement rate of 95.1% (98/103) . When compared to conventional testing kits, we found that the total agreement rate with the Directigen RS (Nippon Becton Dickinson and Company) was 88.3% (91/103) and with RSV TestPack (Dainabot Co., Ltd.) was 91.3% (94/103) . The detection limit of the Directigen Lateral Flow RSV was 2 x 10(3) PFU/ml for both RSV subgroups A and B . In the crossreactivity test, only RSV was found positive . No other microorganisms were crossreactive . We also studied storage stability of nasopharyngeal aspirates and found that stability was not affected by storage at room and refrigerator temperatures for 14 days . Taken all together, the Directigen Lateral Flow RSV is useful for the diagnosis of RSV infection in a clinical setting because its performance is equivalent to conventional testing kits and is easy to use. Biomedica, 2003 Jun, 23(2), 213 - 24 {New fungal classification and their applications in medicine}; Montes B et al.; Fungi are gaining importance with the increased incidence of invasive, often fatal mycoses, in immunocompromised patients . In addition, the increased number of emerging opportunistic pathogens has prompted interest in studies pertaining to fungal classification . The traditional methods of identification and classification of these microorganisms are based on the morphology of the sexual and asexual reproductive structures . Recently, this system of classification has been shown incongruent with approaches that better establish phylogenetic relationships among these organisms . Novel aspects of fungal reclassification are described as follows: 1) the position of fungi in the historical classification systems, 2) the recognition of their polyphyly, based on comparative studies of nucleic acid sequences and on analysis of descent lines and, 3) the contribution of molecular phylogeny to medical mycology, including identification of mycotic agents, their molecular epidemiology, prevention of infection and methods of diagnosis. Genetics, 2003 Jul, 164(3), 1221 - 8 Scaling of mutational effects in models for pleiotropy; Wingreen NS et al.; Mutation-selection models provide a framework to relate the parameters of microevolution to properties of populations . Like all models, these must be subject to test and refinement in light of experiments . The standard mutation-selection model assumes that the effects of a pleiotropic mutation on different characters are uncorrelated . As a consequence of this assumption, mutations of small overall effect are suppressed . For strong enough pleiotropy, the result is a nonvanishing fraction of a population with the "perfect" phenotype . However, experiments on microorganisms and experiments on protein structure and function contradict the assumptions of the standard model, and Kimura's observations of heterogeneity within populations contradict its conclusions . Guided by these observations, we present an alternative model for pleiotropic mutations . The new model allows mutations of small overall effect and thus eliminates the finite fraction of the population with the perfect phenotype. Curr Opin Mol Ther, 2003 Jun, 5(3), 278 - 84 Protein microarray technology for unraveling the antibody specificity repertoire against microbial proteomes; Bacarese-Hamilton T et al.; The genomes of microorganisms responsible for diseases of worldwide medical importance have been sequenced or will be available in the near future . Combinatorial cloning technologies for producing large numbers of proteins have been developed and high-throughput assays such as protein microarrays have been clinically validated for detecting the presence of antibodies directed against microbial antigens in human serum . These scientific and technical achievements offer the opportunity to investigate the natural immune response against the whole proteome of a variety of microorganisms . A powerful combination of genomic information, molecular tools and immunological assays are potentially available to identify the antigens that, either alone or in combination, function as targets of protective immunity, or could be used as markers for serodiagnosis. Rocz Panstw Zakl Hig, 2003, 54(1), 119 - 27 {Fungi growth in buildings}; Nabrdalik M et al.; The results of research into occurrence of fungi in buildings are hereby presented . Indoor air pollution balanced between 10(2)-10(3) cfu/m3 . Examined building surfaces have been found significantly mouldy . Total number of fungi ranged from 3.28 x 10(5) to 9.75 x 10(10) cfu/100 cm2 . The results of the research show the active development of this group of microorganisms . In rooms, over 30 fungi species belonging to 11 genera have been detected . The most frequently occurring ones were fungi genera: Cladosporium, Penicillium, Aspergillus and Acremonium . Among those, potentially toxinogenic species have been separated (Aspergillus ochraceus, Cladosporium cladosporioides, Penicillium chrysogenum, Penicillium notatum) . Apart from micotoxins they have been found to produce huge amounts of conidia which can cause allergy in sensitive people . Hence, people should definitely avoid fungi in their environment. Eur J Biochem, 2003 Aug, 270(15), 3214 - 22 Transglutaminase from Streptomyces mobaraensis is activated by an endogenous metalloprotease; Zotzel J et al.; Streptomyces mobaraensis secretes a Ca2+-independent transglutaminase (TGase) that is activated by removing an N-terminal peptide from a precursor protein during submerged culture in a complex medium {Pasternack, R., Dorsch, S., Otterbach, J . T., Robenek, I . R., Wolf, S . & Fuchsbauer, H.-L . (1998) Eur . J . Biochem . 257, 570-576} . However, an activating protease could not be identified, probably because of the presence of a 14-kDa protein (P14) belonging to the Streptomyces subtilisin inhibitor family . In contrast, if the microorganism was allowed to grow on a minimal medium, several soluble proteases were extracted, among them the TGase-activating protease (TAMEP) . TAMEP was purified by sequential chromatography on DEAE- and Arg-Sepharose and used to determine the cleavage site of TGase . It was clearly shown that the peptide bond between Phe(-4) and Ser(-5) was hydrolyzed, indicating that at least one additional peptidase is necessary to complete TGase processing, even if TAMEP cleavage was sufficient to obtain total activity . Sequence analysis from the N-terminus of TAMEP revealed the close relationship to a zinc endo-protease from S . griseus . The S . griseus protease differs from other members of the M4 protease family, such as thermolysin, in that it may be inhibited by the Streptomyces subtilisin inhibitor . P14 likewise inhibits TAMEP in approximately equimolar concentrations, suggesting its important role in regulating TGase activity. Rhinology, 2003 Jun, 41(2), 76 - 9 Bacteriemia in septoplasty and septorhinoplasty surgery; Kaygusuz I et al.; This study was conducted in an attempt to investigate whether bacteriemia developed in patients with septoplasty and septorhinoplasty in the postoperative period during which an anterior nasal pack was in their nose . Fifty-three patients who went through septoplasty or septorhinoplasty operations were followed in this study . Nasal smear cultures were obtained from all the subjects before the operation . After the packs were retrieved, smears were also obtained from the pack material . Venous blood samples were obtained from the patients immediately before the operation, after the operation and immediately following the retrieval of the pack . When preoperative nasal smear cultures and postoperative pack material cultures of the patients that were obtained at 48 hours were compared, it was seen that different microorganisms were present in 66% of the patients . Bacterial growth was not observed in any of the preoperative blood cultures; whereas 8 patients (15.0%) had bacteriemia in the blood samples obtained immediately after the operation and 9 (16.9%) had growth in the blood samples obtained following the retrieval of the pack . With these results we have seen that bacteriemia can develop in patients with septoplasty and septorhinoplasty . It did not cause serious clinical problems . However, in patients with cardiovascular problems, this possibility should be taken into consideration and relevant preoperative precautions should be implemented. Arch Latinoam Nutr, 2002 Dec, 52(4), 387 - 92 {Use of nopal dietary fiber in a powder dessert formulation}; Saenz C et al.; The development of diverse types of foods of low caloric value and with high content in dietary fiber have occupied a preponderant place in the food industry in the last years, due to the growing interest of the consumers for a healthy and nutritious diet . Pre-cooked or quick to prepare foods are attractive for the time they save; if to this you add their nutritious value, the attractiveness is even greater . For this reason, this study analyzes different formulations of a powder to prepare a dessert (flan), with different percentages of incorporation of nopal flour, as a source of dietary fiber (16%, 18%, 20%) . Two flavors (melon and banana) were tried . It was observed that the flan flavored with banana and with 16% of nopal flour, reached better sensorial characteristics . Greater percentages of nopal flour negatively affected the sensorial characteristics, mainly flavor, color and texture . The analysis showed that the powder presented 5.7% of moisture, low water activity (0.48) and therefore a low total recount of microorganisms . The content of protein was high (27.2%), the ether extract low (2.0%) similar to the caloric contribution (40 Kcal/portion) . The flan showed a 9.8% of total dietary fiber, being greater the contribution of soluble fiber (6.1%) than that of insoluble fiber (3.7%) . Due to these characteristics this formulation could be considered as a food that provides benefits for the human health. J Vet Med Sci, 2003 Jun, 65(6), 727 - 9 Evaluation of the polymorphonuclear cell functions of bottlenose dolphins; Noda K et al.; The functions of polymorphonuclear cells (PMN) are the important non-specific defense mechanisms in the immune system . Especially marine mammals are protected by these mechanisms from the aquatic environment with a large variety of microorganisms . Therefore, we examined the PMN functions of bottlenose dolphins in order to obtain the normal ranges and to standardize the techniques . PMNs were isolated by using lymphocyte isolate solution whose density was 1.077; superoxide production was assessed by nitroblue tetrazolium reduction test (NBT) and phagocytosis was tested by using polystyrene latex beads . We showed that the optimal incubation time was 30 min in NBT assay and 12 hr in phagocytosis assay for dolphin PMNs. Chemosphere, 2003 Sep, 52(9), 1489 - 98 Microbial population of spruce soil in Tatachia mountain of Taiwan; Yang SS et al.; To investigate the role of microorganisms in the ecology and the nutrient transformation of forest soil, soil property, microbial population, biomass and organic acid content of Spruce soil in Tatachia mountain were determined during January 1997 to November 1999 . Soil temperatures were between 5.5 and 15.6 degrees C and soil pH ranged from 3.6 to 5.0 . Total organic carbon and nitrogen contents ranged from 5.83% to 34.35% and from 0.90% to 3.19%, respectively . C/N ratio was between 7.07 and 18.24 . Each gram of dry soil contained microbial biomass carbon 308-870 microg, microbial biomass nitrogen 107-240 microg, malic acid 74-211 nM and succinic acid 32-175 nM . In addition, each gram of dry organic layer contained microbial biomass carbon 216-653 microg, microbial biomass nitrogen 10.3-33.8 microg, formic acid 256-421 nM, acetic acid 301-435 nM, malic acid 795-1027 nM and succinic acid 204-670 nM . About 45.5-90.9% of topsoil samples had higher microbial population than those of subsoil especially in actinomycetes, cellulolytic and phosphate-solubilizing microorganisms . Although rhizosphere of Spruce had higher total organic carbon and total nitrogen content than non-rhizosphere and dwarf bamboo areas, the microbial population had no significant difference among them. Clin Immunol, 2003 Jul, 108(1), 29 - 37 Surfactant protein D (SP-D) serum levels in patients with community-acquired pneumonia small star, filled; Leth-Larsen R et al.; SP-D is a lectin involved in the first line of defense against microorganisms . It is primarily found in the lung but also at extrapulmonary sites and in the circulation . An immunoassay for the quantification of SP-D in serum was established and the SP-D concentration was measured in consecutive blood samples from 61 patients hospitalized for community-acquired pneumonia of suspected bacterial origin . On the day of admission to the hospital the serum SP-D concentration was significantly lower than that in healthy subjects . On day 5, the SP-D concentration had increased on average three times the concentration on admission and then slowly declined toward normal levels . CRP was measured simultaneously but no correlation was observed between concentrations of SP-D and CRP . The results show a wide range of serum SP-D concentration in healthy volunteers and indicate that significant changes occur during pulmonary infection. Cell Microbiol, 2003 Aug, 5(8), 561 - 70 Toll-like receptor (TLR) 2 and TLR4 are essential for Aspergillus-induced activation of murine macrophages; Meier A et al.; Aspergillus fumigatius is a ubiquitous saprophytic fungus that has become the most prevalent airborne fungal pathogen for immunocompromised patients during the last two decades . In this report we have analysed how macrophages recognize this microorganism . Using transfected human HEK 293 cells we demonstrate that NF-kappaB-dependent promoter activation triggered by A . fumigatus is mediated by Toll-like receptors TLR2 and TLR4, whereas no activation was observed in cells overexpressing other distinct TLR proteins (TLR1, TLR3, TLR5-10) . Using macrophages derived from mice lacking TLR2 expression, expressing defective TLR4 or both we found that A . fumigatus conidia and hyphae induce NF-kappaB translocation, release of pro-inflammatory molecules, like TNFalpha, and the chemoattractant MIP-2 in a TLR2- and TLR4-dependent manner . Recognition of A . niger and A . fumigatus, was similar in terms of the parameters analysed, suggesting that pathogenic and non-pathogenic aspergilli are sensed by macrophages in a similar fashion . Finally, we found that recruitment of neutrophils is severely impaired in mice lacking both functional TLR2 and TLR4, but is less impaired in single TLR2- or TLR4-deficient mice, providing evidence that both receptors are required for an optimal immune response to Aspergillus in vivo. Gig Sanit, 2003 Mar-Apr, (2), 34 - 6 {Evaluation of quality of Pacific Ocean squid by biotesting}; Shul'gin IuP; The influence of refrigerator storage on the quality of high-protein raw sea material and its products was studied by using squids as an example . The size of microorganisms in 1 g, the content of nitrogen of volatile bases, that of non-protein nitrogen, and the amount of free amino acids were estimated after cooling and freezing and during storage of frozen squids . In parallel, the relative food value of the cooled and frozen squids and their products were determined by biotesting by means of Tetrachymena pyriformis . Refrigerator storage of squids leads to its muscular tissue degradation whose depth determines the loss of their food value . The results of biotesting of squid meat with the infusoria Tetrachymena pyriformis completely correlate with the data of chemical analysis while assessing the quality. Nature, 2003 Jul 24, 424(6947), 464 - 8 Epub 2003 Jul 13. Crystal structure of human cytochrome P450 2C9 with bound warfarin; Williams PA et al.; Cytochrome P450 proteins (CYP450s) are membrane-associated haem proteins that metabolize physiologically important compounds in many species of microorganisms, plants and animals . Mammalian CYP450s recognize and metabolize diverse xenobiotics such as drug molecules, environmental compounds and pollutants . Human CYP450 proteins CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 are the major drug-metabolizing isoforms, and contribute to the oxidative metabolism of more than 90% of the drugs in current clinical use . Polymorphic variants have also been reported for some CYP450 isoforms, which has implications for the efficacy of drugs in individuals, and for the co-administration of drugs . The molecular basis of drug recognition by human CYP450s, however, has remained elusive . Here we describe the crystal structure of a human CYP450, CYP2C9, both unliganded and in complex with the anti-coagulant drug warfarin . The structure defines unanticipated interactions between CYP2C9 and warfarin, and reveals a new binding pocket . The binding mode of warfarin suggests that CYP2C9 may undergo an allosteric mechanism during its function . The newly discovered binding pocket also suggests that CYP2C9 may simultaneously accommodate multiple ligands during its biological function, and provides a possible molecular basis for understanding complex drug-drug interactions. Ann N Y Acad Sci, 2003 Jun, 990, 12 - 24 Ecology and epidemiology of spotted fever group Rickettsiae and new data from their study in Russia and Kazakhstan; Rudakov NV et al.; Rickettsiae represent a wide range of pathogenicity from classic and new pathogens to endosymbionts of eukaryotic cells . Recent studies of rickettsiae have widened the number of representatives of genus Rickettsia, especially in the spotted fever group (SFG) . Rickettsiae of SFG are tick-borne microorganisms with effective transovarial and transstadial transmission . The main hosts are ticks (Dermacentor, Rhipicephalus, Haemophysalis, Ixodes, and Amlyomma) . Strategy of maintenance of tick microorganisms is vector-type transfer and tropism to endothelial cells or blood cells of animals . The main epidemiological characteristics of SFG rickettsioses are different kinds of anthropogenic activity and connection of morbidity with seasonal tick activity . Two other important characteristics are quantitative and qualitative heterogeneity of its populations (different genotypes of Rickettsia in the same territory and species of ticks, for example) and coexistence of different tick microorganisms (rickettsiae, borreliae, ehrlichiae, tick-borne encephalitis complex viruses, etc.) . The role of new rickettsial genotypes in infectious diseases is poorly understood . Simultaneous study of ticks after bites, blood and skin biopsies of patients may be used for detection of spectrum of tick-borne pathogens in mixed natural foci . Interference between rickettsiae with different virulence may affect its populations and levels of morbidity. Radiat Res, 2003 Aug, 160(2), 198 - 204 Structure of dried cellular alginate matrix containing fillers provides extra protection for microorganisms against UVC radiation; Zohar-Perez C et al.; Soil microorganisms in general and biocontrol agents in particular are very sensitive to UV light . The packaging of biocontrol microorganisms into cellular solids has been developed as a means of reducing loss caused by exposure to environmental UV radiation . The bacterial and fungal biocontrol agents Pantoea agglomerans and Trichoderma harzianum were immobilized in freeze-dried alginate beads containing fillers and subjected to 254 nm UV radiation (UVC) . Immobilization of cells in freeze-dried alginate-glycerol beads resulted in greater survival after UV irradiation than for a free cell suspension . Adding chitin, bentonite or kaolin as fillers to the alginate-glycerol formulation significantly increased bacterial survival . Immobilization in alginate-glycerol-kaolin beads resulted in the highest levels of survival . The transmissive properties of the dried hydrocolloid cellular solid had a major influence on the amount of protection by the cell carrier . Dried alginate matrix (control) transmitted an average of 7.2% of the radiation . Filler incorporation into the matrix significantly reduced UV transmission: Alginate with kaolin, bentonite and chitin transmitted an average of 0.15, 0.38 and 3.4% of the radiation, respectively . In addition, the filler inclusion had a considerable effect on the bead's average wall thickness, resulting in a approximately 1.5- to threefold increase relative to beads based solely on alginate . These results suggest that the degree of protection of entrapped microorganisms against UVC radiation is determined by the UV-transmission properties of the dried matrix and the cellular solid's structure . It is concluded that for maximum protection against UV-radiation-induced cell loss, biocontrol microorganisms should be immobilized in alginate-glycerol beads containing kaolin. J Rheumatol, 2003 Jul, 30(7), 1436 - 9 Increased risk of tuberculosis in patients with rheumatoid arthritis; Carmona L et al.; OBJECTIVE: To quantify the risk of tuberculosis (TB) in an unselected sample of patients with rheumatoid arthritis (RA) compared to the risk in the general population . METHODS: The incidence of TB in the general population of Spain was obtained from the National Network of Epidemiological Surveillance reports . The incidence of TB was ascertained from a cohort of 788 patients with RA selected randomly from the registries of 34 participating centers throughout Spain . A patient was considered a TB case only if information about disease symptoms, microorganism identification, and TB treatment were confirmed in the clinical records . The relative risk of TB in RA was calculated by dividing the standardized mean incidence of TB from 1990 to 2000 in the RA cohort by the mean incidence of TB in Spain during the same years . RESULTS: The mean incidence of TB in the general population of Spain from 1990 to 2000 was 23 cases per 100,000 . Seven cases of TB were identified in the RA cohort, yielding a mean annual incidence (1990-2000) of 134/100,000 patients . The incidence risk ratio of pulmonary TB in patients with RA compared to the general population is 3.68 (95% CI 2.36-5.92) . CONCLUSION: We found a 4-fold increased risk of TB infection in patients diagnosed with RA . These results might help to interpret the magnitude of the problem attributable to the introduction of new therapies in RA. Skin Therapy Lett, 2003 Mar, 8(3), 1 - 4 Cutaneous cleansers; Kuehl BL et al.; Skin cleansers may be an important adjunct to the regimen of those who use cosmetics, have sensitive or compromised skin, or utilize topical therapies . Cleansers emulsify dirt, oil and microorganisms on the skin surface so that they can be easily removed . During cleansing, there is a complex interaction between the cleanser, the moisture skin barrier, and skin pH . Cleansing, with water soap or a liquid cleanser, will affect the moisture skin barrier . Soap will bring about the greatest changes to the barrier and increase skin pH . Liquid facial cleansers are gentler, effecting less disruption of the barrier, with minimal change to skin pH, and can provide people with a cleanser that is a combination of surfactant classes, moisturizers and acidic pH in order to minimize disruption to the skin barrier. Plant Physiol, 2003 Jul, 132(3), 1707 - 15 Identification and differential expression of two thioredoxin h isoforms in germinating seeds from pea; Montrichard F et al.; The NADPH/NADP-thioredoxin (Trx) reductase (NTR)/Trx system (NTS) is a redox system that plays a posttranslational regulatory role by reducing protein targets involved in crucial cellular processes in microorganisms and animals . In plants, the system includes several h type Trx isoforms and has been shown to intervene in reserve mobilization during early seedling growth of cereals . To determine whether NTS was operational during germination of legume seeds and which Trx h isoforms could be implicated, Trx h isoforms expression was monitored in germinating pea (Pisum sativum cv Baccara) seeds, together with the amount of NTR and NADPH . Two new isoforms were identified: Trx h3, similar to the two isoforms already described in pea but not expressed in seeds; and the more divergent isoform, Trx h4 . Active recombinant proteins were produced in Escherichia coli and used to raise specific antibodies . The expression of new isoforms was analyzed at both mRNA and protein levels . The lack of correlation between mRNA and protein abundances suggests the occurrence of posttranscriptional regulation . Trx h3 protein amount remained constant in both axes and cotyledons of dry and imbibed seeds but then decreased 2 d after radicle protrusion . In contrast, Trx h4 was only expressed in axes of dry and imbibed seeds but not in germinated seeds or in seedlings, therefore appearing as closely linked to germination . The presence of NTR and NADPH in seeds suggests that NTS could be functional during germination . The possible role of Trx h3 and h4 in this context is discussed. J Environ Sci Health B, 2003 Jul, 38(4), 479 - 88 Impact of soil management and two botanical insecticides on urease and invertase activity; Antonious GF; Yard waste compost provides an organic amendment useful for improving soil structure and nutrient status . The activities of the enzymes hydrolyzing urea (urease) and sucrose (invertase) in the rhizosphere of potato plants were determined under field conditions . Soil urease and invertase activities were monitored in compost amended soil, grass buffer strips, and in adjacent bare soils during 35 d following soil treatment . Soil urease activity was increased by application of yard waste compost compared to untreated soil which provide evidence of increased soil microbial population following application of compost . Some transitional effects on urease activities were observed following Pyrethrins and Neemix-4E application, these effects were neither drastic nor prolonged enough to be considered deleterious to the soil microorganisms and their activities important to soil fertility . No relationship was found between invertase activity and the three management practices or soil organic matter content . It is suggested that application of botanical insecticides like pyrethrins and Neemix-4E may be useful in delaying urea fertilizer mineralization to maintain N in a form less leachable, so that the duration of N availability to plants is prolonged . The present study may also provide information on urease activity as a sensitive bioindicator of soil quality that reflects the effects of land management on soil quality and may assist land managers in monitoring long-term productivity and sustainability of agricultural lands. J Environ Sci Health B, 2003 Jul, 38(4), 405 - 16 Experimental study on effect of anion surfactant on degradation rate of aldicarb in soil; Liu G et al.; Degradation kinetics of aldicarb {2-methyl-2-(methylthio) propionaldehyde O-(methyl carbamoyl) oxime} in surface and subsurface soil containing different levels of sodium dodecylbenzenesulfonate (SDBS) were determined to understand complex effect of SDBS on aldicarb degradation process . The results showed that degradation curves of aldicarb in soil can be described with first order kinetics formula and the degradation rate constant . k (d(-1)), in surface soil was larger than that in subsurface soil . SDBS can accelerate the degradation of aldicarb in soil and there was a good linear relationship between degradation rate constant and the logarithm of SDBS concentration . The possible reasons were that SDBS could change pH value of soil, have solubilization effect on aldicarb, and be used as carbon source of microorganisms . But SDBS had a larger promotion to the degradation of aldicarb in surface than in subsurface soil . When SDBS concentration was 1000 mg/kg of dried soil the first order degradation rate constant of aldicarb could be increased by 56.6 percent in surface soil and by 27.6 percent in subsurface soil, respectively. Photochem Photobiol, 2003 Jan, 77(1), 34 - 40 Solar UV irradiation conditions on the surface of Mars; Ronto G et al.; The UV radiation environment on planetary surfaces and within atmospheres is of importance in a wide range of scientific disciplines . Solar UV radiation is a driving force of chemical and organic evolution and serves also as a constraint in biological evolution . In this work we modeled the transmission of present and early solar UV radiation from 200 to 400 nm through the present-day and early (3.5 Gyr ago) Martian atmosphere for a variety of possible cases, including dust loading, observed and modeled O3 concentrations . The UV stress on microorganisms and/or molecules essential for life was estimated by using DNA damaging effects (specifically bacteriophage T7 killing and uracil dimerization) for various irradiation conditions on the present and ancient Martian surface . Our study suggests that the UV irradiance on the early Martian surface 3.5 Gyr ago may have been comparable with that of present-day Earth, and though the current Martian UV environment is still quite severe from a biological viewpoint, we show that substantial protection can still be afforded under dust and ice. Photochem Photobiol Sci, 2002 Jun, 1(6), 431 - 7 A quantitative method for evaluating the photoreactivation of ultraviolet damaged microorganisms; Beggs CB; The lethal effect of ultraviolet (UV) light on microorganisms is well known and many studies have been undertaken into the effects of UV induced damage . Most of this work has been experimental; by comparison relatively little theoretical work has been undertaken to analyse the kinetics of the UV inactivation process, or to develop quantitative methodologies to support the experimental work . This paper presents a new and simple model for quantifying the photolysis rate . A theoretical study is also presented in this paper which quantifies photolysis rates for E . coli O26 and E . coli O157:H7 . This study uses experimental data collected by Tosa and Hirata, and reveals the photolysis rate for E . coil O26 during the UV irradiation process to be 4.69 x 10(-3) m2 J(-1) . By comparison, E . coli O157:H7 is much more susceptible to UV induced damage than E . coli O26, having a photolysis constant of only 2.09 x 10(-3) m2 J(-1). FEMS Microbiol Lett, 2003 Jul 15, 224(1), 127 - 32 Vitreoscilla hemoglobin promoter is not responsive to nitrosative and oxidative stress in Escherichia coli; Frey AD et al.; The Vitreoscilla hemoglobin gene (vhb) is expressed under oxygen-limited conditions via an FNR-dependent mechanism . Furthermore, cAMP-CRP has been implicated in its regulation . Recently, VHb protein has been reported to protect a heterologous host from nitrosative stress . In this study we analyzed the regulation of the Vitreoscilla hemoglobin promoter (Pvhb) in Escherichia coli under nitrosative and oxidative stress conditions . Our results show unambiguously that expression of neither VHb nor chloramphenicol acetyltransferase under the control of Pvhb is induced under the experimental conditions used . Thus, a clear discrepancy between in vivo function, i.e . protection against nitrosative stress, and regulation of gene expression is obvious . The regulation of Pvhb reported here is in clear contrast to the expression pattern of flavohemoglobins from various microorganisms, which are generally induced by nitrosative stress . However, the length of Pvhb is only 146 bp and therefore, we cannot rule out that additional regulatory sequences may be located in the upstream region of Pvhb. Bull Mem Acad R Med Belg, 2002, 157(10-12), 479 - 84 {Hereditary abnormalities in the immune system: physiology and treatment}; Fischer A; There are multiple genetic abnormalities of the immune system . About one hundred syndromes have been so far described . All of them disturb immune defenses against microorganism invasions and/or the homeostasis of the immune system . Thanks to the advances in genomics, genetic and molecular basis of more than 80 of these conditions, have been unravelled . These achievements provide significant insight into the understanding of many steps of the biology of the immune system, i.e., its development, function and regulation . It is also the basis for the design of molecular diagnostic tools and perhaps for new therapeutics of these severe disorders. Nucleic Acids Res, 2003 Jul 15, 31(14), 4024 - 30 Processing of DNA lesions by archaeal DNA polymerases from Sulfolobus solfataricus; Gruz P et al.; Spontaneous damage to DNA as a result of deamination, oxidation and depurination is greatly accelerated at high temperatures . Hyperthermophilic microorganisms constantly exposed to temperatures exceeding 80 degrees C are endowed with powerful DNA repair mechanisms to maintain genome stability . Of particular interest is the processing of DNA lesions during replication, which can result in fixed mutations . The hyperthermophilic crenarchaeon Sulfolobus solfataricus has two functional DNA polymerases, PolB1 and PolY1 . We have found that the replicative DNA polymerase PolB1 specifically recognizes the presence of the deaminated bases hypoxanthine and uracil in the template by stalling DNA polymerization 3-4 bases upstream of these lesions and strongly associates with oligonucleotides containing them . PolB1 also stops at 8-oxoguanine and is unable to bypass an abasic site in the template . PolY1 belongs to the family of lesion bypass DNA polymerases and readily bypasses hypoxanthine, uracil and 8-oxoguanine, but not an abasic site, in the template . The specific recognition of deaminated bases by PolB1 may represent an initial step in their repair while PolY1 may be involved in damage tolerance at the replication fork . Additionally, we reveal that the deaminated bases can be introduced into DNA enzymatically, since both PolB1 and PolY1 are able to incorporate the aberrant DNA precursors dUTP and dITP. Proc Natl Acad Sci U S A, 2003 Jul 22, 100(15), 9061 - 6 Epub 2003 Jul 09. Identification of histidine kinases that act as sensors in the perception of salt stress in Synechocystis sp . PCC 6803; Marin K et al.; In plants and microorganisms, salt stress regulates the expression of large numbers of genes . However, the machinery that senses salt stress remains to be characterized . In this study we identified sensory histidine kinases that are involved in the perception of salt stress in the cyanobacterium Synechocystis sp . strain PCC 6803 . A library of strains with mutations in all 43 histidine kinases was screened by DNA microarray analysis of genomewide gene expression under salt stress . The results suggested that four histidine kinases, namely, Hik16, Hik33, Hik34, and Hik41, perceived and transduced salt signals . However, Hik33, Hik34, and Hik16 acting with Hik41 regulated the expression of different sets of genes . These histidine kinases regulated the expression of approximately 20% of the salt-inducible genes, whereas the induction of the remaining salt-inducible genes was unaffected by mutations in any of the histidine kinases, suggesting that additional sensory mechanisms might operate in the perception of salt stress . We also used DNA microarrays to investigate the effect of various salts on gene expression . Our results indicate that Hik33 responds to sodium salts and not to KCl, whereas the Hik16/Hik41 system responds only to NaCl. Ann Agric Environ Med, 2003, 10(1), 9 - 14 In vitro toxicity of indoor Chaetomium Kunze ex Fr; Pieckova E; Microscopic fungi in the indoor environment present a serious health risk for people living in affected buildings . The potentially toxic ascomycete genus Chaetomium is supposed to be the third most frequent indoor fungal contaminant . Its brief mycological, toxicological and ecological characterization is given . The work was aimed at in vitro study of toxicity of endo- and exometabolites of 14 strains of Chaetomium spp., including 4 strains of Ch . globosum, isolated from mouldy buildings in Slovakia and Denmark, and 3 Ch . globosum strains from the Czechoslovak Collection of Microorganisms (CCM) . The endometabolites of 10 isolates of Chaetomium spp . were active: 7 isolates (41% of total strain number) stopped tracheal ciliary movement of 1-d-old chickens after 24 h, 9 isolates (53%) after 48 h and 10 strains (59%) after 72 h . In the case of exometabolites, the extracts of 6 Chaetomium strains showed some ciliostatic activity: 2 isolates (12% of strains tested) after 24 h, 5 isolates (29%) after 48 h and 6 isolates (35%) after 72 h . In general, 5 isolates of Danish origin (83%) produced ciliostatically active exometabolites and 2 isolates (33%) produced such endometabolites, while only 4 strains isolated in Slovakia (50%) and 3 strains (37%) respectively did the same under experimental conditions . Most toxic metabolites were produced by Chaetomium spp . isolated from dwellings, whereas hospital isolates were not able to produce active compounds . Chaetomia as indoor contaminants can contribute to ill health of occupants of mouldy damp buildings. Glycobiology, 2003 Oct, 13(10), 93R - 104R Epub 2003 Jul 08. Glycosidase inhibitors: update and perspectives on practical use; Asano N; About 40 years have passed since the classical glycosidase inhibitor nojirimycin was discovered from the cultured broth of the Streptomyces species . Since then, over 100 glycosidase inhibitors have been isolated from plants and microorganisms . Modifying or blocking biological processes by specific glycosidase inhibitors has revealed the vital functions of glycosidases in living systems . Because enzyme-catalyzed carbohydrate hydrolysis is a biologically widespread process, glycosidase inhibitors have many potential applications as agrochemicals and therapeutic agents . Glycosidases are involved in the biosynthesis of the oligosaccharide chains and quality control mechanisms in the endoplasmic reticulum of the N-linked glycoproteins . Inhibition of these glycosidases can have profound effects on quality control, maturation, transport, and secretion of glycoproteins and can alter cell-cell or cell-virus recognition processes . This principle is the basis for the potential use of glycosidase inhibitors in viral infection, cancer, and genetic disorders . In this review, the past and current applications of glycosidase inhibitors to agricultural and medical fields and the prospect for new therapeutic applications are reconsidered. Curr Opin Biotechnol, 2003 Jun, 14(3), 303 - 10 Biotechnological prospects from metagenomics; Schloss PD et al.; The recognition that most microorganisms in the environment cannot be cultured by standard methods stimulated the development of metagenomics, which is the genomic analysis of uncultured microorganisms . Two types of analysis have been used to obtain information from metagenomic libraries: a function-driven approach, in which metagenomic libraries are initially screened for an expressed trait, and a sequence-driven approach, in which libraries are initially screened for particular DNA sequences . New antibiotics and enzymes are among the early discoveries from metagenomics . Future refinement of methods that enrich for genes of particular function will accelerate the rate of discovery of useful molecules. Curr Opin Biotechnol, 2003 Jun, 14(3), 296 - 302 Stable-isotope probing of nucleic acids: a window to the function of uncultured microorganisms; Radajewski S et al.; Phylogeny based on ribosomal RNA sequences alone is rarely a reliable indicator of microbial function . To circumvent this problem, nucleic acid based techniques have been developed that exploit the physical properties of stable isotopes to study microbially mediated processes within complex environmental samples . Investigations using labelled substrates, or which detect variations in the natural abundance of isotopes, have thus revealed the metabolic function of microorganisms without the need to isolate them in culture. Dent Clin North Am, 2003 Jul, 47(3), 545 - 57 Waterborne pathogens and dental waterlines; Mills SE; Humans, like every other living thing on Earth, have evolved in a world dominated by many billions of microscopic life forms . Most of the time, we live in a state of harmony (or even mutualism) with our invisible coinhabitants . When this balance becomes disturbed however, the consequences can be devastating . Infectious diseases including malaria, tuberculosis, and AIDS remain the world's greatest mass murderers . Dental workers strive to reduce infection risks for their patients through infection control measures that reduce or eliminate potentially pathogenic agents in the clinical environment . As increasing numbers of patients with varying degrees of immune suppression present for dental treatment, the need to ensure an aseptic treatment environment will become a higher priority for the dental profession . The possibility that exposure to aerosols contaminated with endotoxin might exacerbate asthma or cause chronic respiratory problems in dental health care workers should be investigated . Although direct evidence of widespread complications among patients or occupationally acquired illness among dental workers is presently lacking, reducing the numbers of microorganisms present in dental treatment water is consistent with other empiric measures that form the basis of infection-control practice. Crit Care Med, 2003 Jul, 31(7), 1938 - 46 Fluconazole improves survival in septic shock: a randomized double-blind prospective study; Jacobs S et al.; OBJECTIVE: To demonstrate whether fluconazole reduces multiple organ failure and mortality in early septic shock (<24 hrs) . DESIGN: A prospective randomized double-blind study . SETTING: A medical and surgical adult intensive care unit in a tertiary referral center . PATIENTS: Values were obtained from 71 general adult intensive care unit patients . INTERVENTIONS: During a 2.5-yr period, December 1998-June 2001, 71 patients with septic shock attributed to either nosocomial pneumonia (n = 37) or intra-abdominal sepsis (n = 34) were admitted to our intensive care unit and met the criteria of early septic shock and were entered into this study . All patients were randomized by our clinical pharmacist to receive daily either 200 mg of fluconazole in isotonic saline (fluconazole group = 32) or isotonic saline alone (placebo group = 39) intravenously during the course of their septic shock . MEASUREMENTS AND MAIN RESULTS: All patients were closely monitored with pulmonary artery catheters and parameters to calculate daily organ dysfunction and Acute Physiology and Chronic Health Evaluation II scores . There was a highly significant increase in 30-day survival in the fluconazole-treated patients compared with the placebo patients (78% vs . 46%) . However, fluconazole was found to be more effective in patients with septic shock attributed to intra-abdominal sepsis than to nosocomial pneumonia . Increased survival in the intra-abdominal sepsis clinical category was mirrored by a significantly lower number of organ failures in the treated group compared with the placebo group whereas the number of organ failures in the fluconazole group attributed to nosocomial pneumonia were not significantly increased compared with the control group . The septic shock state was considered in all cases to be attributed to bacterial and not to disseminated yeast infection with the exception of one patient in the control group who was admitted with candidemia . The mechanisms by which fluconazole exerts its protective effect against septic shock in patients is far from clear . However, fluconazole has been shown to enhance bactericidal activity of neutrophils and also to inhibit transmigration and adhesion of neutrophils in capillaries of distant organs . CONCLUSIONS: The development of organ failure and mortality in septic shock was significantly reduced by fluconazole given intravenously . The mechanism of action of fluconazole in reducing multiple organ dysfunction in this group of patients may be attributed to the ability of fluconazole to increase recruitment, improve bactericidal activity of neutrophils, and to contain microorganisms locally. Curr Treat Options Gastroenterol, 2003 Aug, 6(4), 283 - 288 Probiotics, Irritable Bowel Syndrome, and Inflammatory Bowel Disease; Floch MH; Probiotics are live, microbial food supplements that benefit the host animal by improving intestinal microbial balance . Their major role in preventing and treating gastrointestinal disease appears to be from their effect on the immune process, protection against abnormal invasive bacteria, and in the production of short-chain fatty acids from starch and non-starch polysaccharides . Probiotic microorganisms are administered in food supplements and yogurts . They are also now sold in the form of capsules and powders . There is great variation in the microorganisms in the various supplements . It is important to understand that all probiotic products are different . Some contain a single organism and others contain multiple organisms . Therapeutic results have been achieved with various probiotics in different diseases . In the treatment of inflammatory bowel diseases (IBD), success has been reported with Escherichia coli Nissle strain in ulcerative colitis, and with a multiple organism product, VSL#3 (VSL Pharmaceuticals, Fort Lauderdale, FL), in Crohn's disease and pouchitis . Initial reports in irritable bowel syndrome (IBS) have resulted in encouraging results with the use of E . coli Nissle strain, and recently with multiple organism probiotic supplements . However, caution must still apply to the use of probiotics in IBD and IBS because the reports and the number of patients treated are limited. Kidney Int, 2003 Aug, 64(2), 572 - 8 Effect of iron treatment on circulating cytokine levels in ESRD patients receiving recombinant human erythropoietin; Weiss G et al.; BACKGROUND: Anemia in patients with end-stage renal disease (ESRD) is treated with recombinant human erythropoietin (rhEPO) often in combination with iron . However, iron catalyzes the formation of toxic radicals, which might promote vascular damage, is a nutrient for microorganisms, and negatively affects immune pathways, thus increasing the risk for severe infections . METHODS: We investigated 28 patients on chronic hemodialysis who were randomized to receive either rhEPO alone (N = 15) or rhEPO in combination with intravenous iron (N = 13) for a period of 12 weeks . We analyzed iron therapy associated changes in cytokine patterns and endogenous radical formation . RESULTS: Tumor necrosis factor-alpha (TNF-alpha) levels were increased in ESRD patients at study entry and then decreased significantly over time in subjects receiving additional iron, while they increased with rhEPO alone . In contrast, we found serum concentrations of the anti-inflammatory cytokine interleukin (IL)-4 to increase with iron therapy . A significant negative correlation between iron availability, as determined by transferrin saturation, and TNF-alpha levels (P = 0.008) and a positive one between transferring saturation and IL-4 (P = 0.02) pointed to the potential role of iron to induce immunologic changes . Interestingly, iron therapy resulted in a slight decrease in the amounts of endogenous peroxides, which may be referred to reduced TNF-alpha concentrations since peroxide concentrations were positively correlated to TNF-alpha levels (P = 0.046) and negatively to transferrin saturation (P = 0.02) . CONCLUSION: Iron supplementation in ESRD patients down-regulates proinflammatory immune effector pathways and stimulates the expression of the anti-inflammatory cytokine IL-4 . Such a condition is detrimental for host response toward invading pathogens . However, tissue damage by radicals such as endogenous peroxides may be reduced in this condition due to impaired TNF-alpha formation. Ann Pharm Fr, 2003 Jul, 61(4), 270 - 5 {Bacterial aggression}; Berche P; In all living species, the first line of defence against microbial aggressions is constituted by innate immunity . During Evolution, it appears in invertebrates and plants, long before adaptive immunity, which appears in vertebrate . Adaptive immunity induces acquired resistance against microorganisms through random somatic rearrangements of genes encoding immunoglobulins and T cell receptors, thus generating a high level of diversity of receptors (>10(9)) in response to microbial aggressions . Acquired resistance is not vertically transmitted and reflects the "infectious history" of every individual . In contrast, innate immunity relies on recognition of antigens by a small number of weakly specific receptors (>10(2)) designated Pattern-Recognition Receptors (PRR) and is vertically transmitted by germinal cells . The PRR are expressed on macrophages dendritic cells and B lymphocytes and recognize antigenic structures highly conserved in the living world, termed Pathogen-Associated Molecular Patterns (PAMP), as lipopolysaccharides peptidoglycanes and lipoteichoic acids . PRR are secreted (complement, lectins), or expressed at the cell surface of cells to induce endocytosis or signaling (Toll-like receptors or TLRs) . The recognition of antigens induces an immediate inflammatory response and triggers adaptive immunity . Among secreted PRR, the system of complement plays a major role in the immediate inflammatory response, controlling infections by its major role in opsonization, chemotactism and activation of leucocytes . TLRs induce the inflammatory response against microorganisms through NF-kB, a cytoplasmic factor controlling transcription of many genes, including cytokines (TNF, INF, IL-1, IL-2, IL-8, IL-12.) and defensines . So, within few minutes following microbial aggression, the inflammatory response is rapidly triggered to destroy infectious agents and to generate a long-term memory against pathogens. J Neurosci, 2003 Jul 2, 23(13), 5536 - 44 Glucocorticoids play a fundamental role in protecting the brain during innate immune response; Nadeau S et al.; The innate immune system plays a crucial role in protecting the host against infectious microorganisms . An inappropriate control of this system may have profound consequences, because of the maintained production of specific proinflammatory molecules . Glucocorticoids are the most efficient endogenous molecules that provide negative feedback on proinflammatory signaling and gene expression . Here we show that activation of this system is not detrimental for the brain but a profound neurodegeneration takes place in animals treated with the glucocorticoid receptor inhibitor Mifepristone (RU486) . This drug increased the inflammatory reaction induced by a single intracerebral bolus of lipopolysaccharide (LPS) . Inhibition of tumor necrosis factor alpha (TNF-alpha) totally abolished the neurotoxic effect of the endotoxin, and chronic infusion of the cytokine mimicked the treatment combining RU486 and LPS . The neuronal damage caused by TNF-alpha is dependent on both nitric oxide and caspase pathways . In controlling the cerebral innate immunity and microglial TNF-alpha production, glucocorticoids play a major role in protecting the brain against bacterial cell wall components. Forensic Sci Int, 2003 Jun 24, 134(1), 62 - 71 Microbial degradation of illicit drugs, their precursors, and manufacturing by-products: implications for clandestine drug laboratory investigation and environmental assessment; Janusz A et al.; Chemicals associated with clandestine drug laboratories are often disposed of covertly into soil, sewerage systems, or public waste management facilities . There are two significant issues relating to such dumps of materials; they might contain valuable evidence as to drug manufacture, and they might be a source of pollution . This study presents initial findings in relation to the impact microorganisms from environmental sources have upon drugs, their precursors, and manufacturing by-products . The aim of this study was to identify which chemicals associated with clandestine drug laboratories persist in the environment in order to allow forensic drug chemists to link discarded residues with the method of manufacture, and to allow the environmental impact of clandestine drug laboratories to be assessed accurately . When exposed to soil microorganisms, phenyl-2-propanone (P2P) was rapidly metabolized into mixtures of 1-phenyl-2-propanol, 1-phenyl-1,2-propanedione, 1-hydroxy-1-phenyl-2-propanone, 2-hydroxy-1-phenyl-1-propanone, and the two diastereoisomers of 1-phenyl-1,2-propanediol . On the other hand, when exposed under the same conditions, methylamphetamine sulphate (MAS) remained virtually unchanged . Implications relating to evidence gathering for forensic purposes and to environmental assessment of clandestine drug laboratories are discussed. Semin Respir Infect, 2003 Jun, 18(2), 103 - 11 Evaluation of nonresolving and progressive pneumonia; Menendez R et al.; The concepts of nonresolving and progressive pneumonia are difficult to define: both refer to a failure in the therapeutic response, which in the case of progressive pneumonia may cause a medical emergency even in the first 72 hours after empiric treatment . The incidence of nonresolving pneumonia in community-acquired pneumonia is approximately 10%, and greater than 30% in nosocomial pneumonia . Mortality in nonresponding pneumonia increases 3-fold in community-acquired and 5-fold in nosocomial pneumonia compared with global mortality in hospitalized patients . Factors associated with the resolution of pneumonia are related to the host, the microorganisms, and the cytokine response that modulates the relationship between them . Causes of nonresolving or progressive pneumonia may be infectious or noninfectious . Management of nonresponding patients requires a reevaluation of epidemiologic data, a complete microbiologic investigation, with conventional and invasive respiratory samples, and performance of a new radiographic study . Empiric therapeutic changes are aimed at broadening bacteriologic coverage to treat resistant or unusual microorganisms. Appl Environ Microbiol, 2003 Jul, 69(7), 4249 - 55 Detection and diversity assessment of Xylella fastidiosa in field-collected plant and insect samples by using 16S rRNA and gyrB sequences; Rodrigues JL et al.; The causal agent of diseases in many economically important plants is attributed to the xylem-limited bacterium Xylella fastidiosa . The detection of this plant pathogen has been hampered due to its difficult isolation and slow growth on plates . Nearly complete nucleotide sequences of the 16S rRNA gene and partial sequences of the gyrB gene were determined for 18 strains of X . fastidiosa isolated from different plant hosts . A phylogenetic analysis, based on gyrB, grouped strains in three clusters; grape-isolated strains formed one cluster, citrus-coffee strains formed another cluster, and a third cluster resulted from all other strains . Primer pairs designed for the 16S rRNA and gyrB genes were extensively searched in databases to verify their in silico specificity . Primer pairs were certified with 30 target and 36 nontarget pure cultures of microorganisms, confirming 100% specificity . A multiplex PCR protocol was developed and its sensitivity tested . Sequencing of PCR products confirmed the validity of the multiplex PCR . Xylella fastidiosa was detected in field-collected plants, disease vector insects, and nonsymptomatic but infected plants . Specific detection of X . fastidiosa may facilitate the understanding of its ecological significance and prevention of spread of the disease. Appl Environ Microbiol, 2003 Jul, 69(7), 3858 - 67 Microbial diversity of cryptoendolithic communities from the McMurdo Dry Valleys, Antarctica; de la Torre JR et al.; In the McMurdo Dry Valleys of Antarctica, microorganisms colonize the pore spaces of exposed rocks and are thereby protected from the desiccating environmental conditions on the surface . These cryptoendolithic communities have received attention in microscopy and culture-based studies but have not been examined by molecular approaches . We surveyed the microbial biodiversity of selected cryptoendolithic communities by analyzing clone libraries of rRNA genes amplified from environmental DNA . Over 1,100 individual clones from two types of cryptoendolithic communities, cyanobacterium dominated and lichen dominated, were analyzed . Clones fell into 51 relatedness groups (phylotypes) with > or =98% rRNA sequence identity (46 bacterial and 5 eucaryal) . No representatives of Archaea were detected . No phylotypes were shared between the two classes of endolithic communities studied . Clone libraries based on both types of communities were dominated by a relatively small number of phylotypes that, because of their relative abundance, presumably represent the main primary producers in these communities . In the lichen-dominated community, three rRNA sequences, from a fungus, a green alga, and a chloroplast, of the types known to be associated with lichens, accounted for over 70% of the clones . This high abundance confirms the dominance of lichens in this community . In contrast, analysis of the supposedly cyanobacterium-dominated community indicated, in addition to cyanobacteria, at least two unsuspected organisms that, because of their abundance, may play important roles in the community . These included a member of the alpha subdivision of the Proteobacteria that potentially is capable of aerobic anoxygenic photosynthesis and a distant relative of Deinococcus that defines, along with other Deinococcus-related sequences from Antarctica, a new clade within the Thermus-Deinococcus bacterial phylogenetic division. J Invest Dermatol, 2003 Jul, 121(1), 28 - 36 Human glands of Moll: histochemical and ultrastructural characterization of the glands of Moll in the human eyelid; Stoeckelhuber M et al.; The function of the human gland of Moll of the eyelid is not exactly known . We studied the secretory and cytoskeletal components of these apocrine glands in males and females by immunohistochemical methods, and the ultrastructural organization of the glandular cells with an electron microscope . The glands of Moll are exclusively located at the margin of the eyelids and their ducts empty into the lash follicle . Immunohistochemical staining for actin and cytokeratins CK19 and CK7 points to the involvement of actin in the pinching-off mechanism of the apical cell protrusion during apocrine secretion and to a stabilizing role for the cytokeratins in this apical region of the glandular cells . The presence of the bacteriolytic enzyme lysozyme, the membrane-associated mucin 1, and the immunoglobulin A and its secretory component within the gland suggest a function in local immune defense . The presence of a variety of sugar components in the secretory product was verified by lectin histochemistry and periodic acid Schiff and Alcian blue stain . We suppose that these apocrine glands are active from birth in producing agents against pathogenic microorganisms in the eyelid shaft and on the ocular surface. Res Microbiol, 2003 Jun, 154(5), 315 - 20 Response of terrestrial microorganisms to ultraviolet-B radiation in ecosystems; Johnson D; Recent studies suggest that ultraviolet-B radiation, a major component of global climate change, can affect the community structure, biomass and functioning of terrestrial microorganisms . In this report, these studies are reviewed and gaps in our understanding highlighted so that future research can provide more conclusive evidence on the mechanisms and wider ecological implications of the impacts of elevated ultraviolet-B radiation on terrestrial microorganisms. J Dairy Sci, 2003 Jun, 86(6), 1932 - 40 Effects of carbon dioxide on bacterial growth parameters in milk as measured by conductivity; Martin JD et al.; Inhibition of bacterial growth by dissolved carbon dioxide (CO2) has been well established in many foods including dairy foods . However, the effects of dissolved CO2 on specific growth parameters such as length of lag phase, time to maximum growth rate, and numbers of organisms at the stationary phase have not been quantified for organisms of concern in milk . The effect of dissolved CO2 concentrations of 0.6 to 61.4 mM on specific bacterial growth parameters in raw or single organism inoculated sterile milk was determined at 15 degrees C by conductance . Commingled raw or sterile milks were amended to a final concentration of 0.5 mg/ml each of urea and arginine HCl . Sterile milks were inoculated singly with one of six different microorganisms to a final concentration of approximately 10(2) to 10(3) cfu/ml; raw milk was adjusted to a final indigenous bacterial population of approximately 10(3) cfu/ml . Conductivity of the milk was recorded every 60 s over 4 to 5 d in a circulating apparatus at 15 degrees C . Conductivity values were fit to Gompertz equations and growth parameters calculated . Conductance correlated with plate counts and was satisfactory for monitoring microbial growth . Data fit the Gompertz equation with high correlation (R2 = 0.96 to 1.00) . In all cases, dissolved CO2 significantly inhibited growth of raw milk bacteria, influencing lag, exponential, and stationary growth phases as well as all tested monocultures. Ying Yong Sheng Tai Xue Bao, 2003 Mar, 14(3), 457 - 60 {A review on phytoremediation of organic contaminants}; Xia H et al.; Plants can take up and accumulate most persistant chemicals such as PCBs and PAHs by roots . Some chemicals such as TNT, TCE, and most organic pesticides can be phytodegraded into polar metabolites, or assimilated to CO2 and water by some plants . On the other hand, more microorganisms in rhizosphere can enhance the degradation rate of organic contaminants in environment . The benefits and some issues on phytoremediation are also discussed. Ying Yong Sheng Tai Xue Bao, 2003 Mar, 14(3), 453 - 6 {Advances in extraction and purification of soil microorganism}; Xiang W et al.; For the studies on soil microbial ecology and bio-diversity as well as on the dynamics of C, N, S and P in soil, it is necessary to quantitatively extract microorganism from soil . The recent advances on extraction and purification of microorganism from soil were reviewed in this paper . The procedure for extracting microorganism from soil was comprised of three steps, i.e., soil dispersion, separation and purification . Most bacteria in soil could be successively extracted from soil particles by present techniques such as filtration, centrifugation and elutriation . However, filamentous fungi seem more difficult to be extracted than bacteria, because it is tangled with soil particles . Rotating wire frame, aqueous extraction and membrane filter technique, and low-speed centrifugation were used for extracting filamentous fungi from soil, but the extraction efficiency was rather low and needed to be raised . Aqueous two-phase partitioning system was successfully adopted as a technique for bacteria purification, and it would possibly become a technique suitable for fungi purification . The potential area of research on soil microbial ecology as well as on soil and plant nutrition by using microbial extraction technique was also discussed. J Orofac Orthop, 2003 May, 64(3), 214 - 27 Movement of periodontally affected teeth after guided tissue regeneration (GTR)--an experimental pilot study in animals; Diedrich P et al.; PURPOSE: The purpose of the present experimental animal study was to examine the relevance of regenerative periodontal therapy (membrane procedure, application of enamel matrix proteins) to subsequent orthodontic tooth movement . MATERIAL AND METHOD: After extraction of the third premolars in four foxhounds, 64 three-wall bony defects were artificially generated at the first and second premolars and colonized by periodontopathogenic microorganisms . After 6 months, regenerative periodontal therapy was performed: scaling, conditioning of the root surfaces with citric acid, combination of enamel matrix protein (Emdogain, Biora, Malmo, Sweden) and resorbable Vicryl membrane (Ethicon, Norderstedt, Germany) . The defects at the first premolars served as the control group . In the test group eight second premolars were intruded 1 month after surgical intervention; in addition eight premolars were translatorily distalized, i.e . moved both into the defect (pressure side) and away from the defect (tension side) . Qualitative histologic analysis was performed on non-decalcified microsection samples using the polychrome sequential labeling and histomorphometry . RESULTS: The histologic findings revealed extensive periodontal regeneration in the control group, at the intruded root segments, and on the tension sides . Cemento- and osseoneogenesis yielded mean values of 70-80%, whereas values for epithelial down-growth were low . The bony regeneration on the pressure side was markedly reduced; however, nearly 70% new formation of root cementum with Sharpey's fibers was also recorded . CONCLUSION: Periodontal regenerative procedures appear to enhance conditions for the movement of teeth with attachment loss; clinical testing is promising. Rev Argent Microbiol, 2003 Jan-Mar, 35(1), 8 - 18 {Prediction of the bacteriological quality of natural waters in urban environments . Effect of the El Niño-Southern Oscillation}; Emiliani F et al.; At the moment of examination, the bacteriological quality of water is not, to a large extent, more than an a posteriori evaluation: the population is advised afterwards that it had been exposed to a sanitary risk . The ideal situation for managing aquatic environments or recreational use would be to be able to predict at any moment their bacteriological quality . An approach would be to develop predictive models to relate the exceedance (number of days during which the bacteriological indicator exceeds the standards) with environmental variables . Studies were carried out at two beaches with different limnological characteristics, a fluvial beach and a lacustrine beach, from the beginning of 1998 to the end of 2000, so that it was possible to obtain information on the impact of very different hydroclimatic situations on microorganisms of sanitary interest (Escherichia coli and cyanobacteria) . Three variables, maximum concentration of E . coli, rains and water level, justified 92.6% of the exceedance variance in the multiple regression model at the fluvial beach . Other abiotic variables (temperature, pH, conductivity, transparency and dissolved oxygen) showed collinearity problems or did not make statistically significant contributions to the model . During the El Nino event (gauge heights > 5 m) there were no exceedances after the rains, even with precipitations of 200 mm . Moreover, cyanobacteria density remained low (x = 1.191 +/- 0.703 log10 cian./ml) . During the negative pluvial anomalies, although the exceedances of E . coli were not very frequent at both beaches due to the scarce rains, a massive development of cyanobacteria was produced (x = 4.657 +/- 0.578 log10 cian./ml) at the lacustrine beach . At this beach, the mean decay coefficient (K) for E . coli was significantly (p < 0.05) lower than that estimated for the fluvial beach (K = -0.682 +/- 0.32 versus K = -0.387 +/- 0.16, respectively), possibly due to the unfavourable environment created by the higher development of photosynthetic organisms . Analizing separately the anomalous period, variables that better explained (98.6%) the exceedances were (in addition to the maximum concentration of E . coli) pH, temperature and days without rains (DWR) before the sampling . The longest exceedances (6 to 8 days) were produced at the end of the negative pluvial anomalies, after prolonged periods (> 30 days) of droughts . Regression models obtained to predict the exceedance of E . coli were simplified using only three abiotic variables (water level, rains and DWR) . Although, in this way, the percentage of explanation of the exceedance is lower (78.5%), the information needed for the model is obtained quickly and preventive measures can be carried out immediately while waiting for more information. J Gen Appl Microbiol, 2003 Apr, 49(2), 63 - 73 Growth inhibition of the filamentous fungus Aspergillus nidulans by cadmium: an antioxidant enzyme approach; Guelfi A et al.; The heavy metal cadmium is very toxic to biological systems . Although its effect on the growth of microorganisms and plants has been investigated, the response of antioxidant enzymes of Aspergillus nidulans to cadmium is not well documented . We have studied the effect of cadmium (supplied as CdCl(2)) on catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR) . 0.005 mM CdCl(2) had a very slight stimulatory effect on the growth rate of A . nidulans, but at concentrations above 0.025 mM, growth was totally inhibited . The accumulation of Cd within the mycelium was directly correlated with the increase in the concentration of CdC(2) used in the treatments . Although a cadmium-stimulated increase in SOD activity was observed, there was no change in the relative proportions of the individual Mn-SOD isoenzymes . Higher concentrations of CdCl(2) induced a small increase in total CAT activity, but there was a major increase in one isoenzymic form, that could be separated by gel electrophoresis . GR activity increased significantly following treatment with the highest concentration (0.05 mM) of CdCl(2) . The increases in SOD, CAT, and GR activities suggest that CdCl(2) induces the formation of reactive oxygen species inside the mycelia of A . nidulans. Mol Biol Evol, 2003 Sep, 20(9), 1545 - 55 Epub 2003 Jun 27. Increased rates of sequence evolution in endosymbiotic bacteria and fungi with small effective population sizes; Woolfit M et al.; Mutualistic, maternally transmitted endosymbiotic microorganisms undergo severe population bottlenecks at each host generation, resulting in a reduction in effective population size (Ne) . Previous studies of Buchnera, the primary endosymbiont of aphids, and of several other species of endosymbiotic bacteria have shown that these species exhibit an increase in the rate of substitution of slightly deleterious mutations, among other predicted effects of increased drift due to small Ne, such as reduced codon bias . However, these studies have been limited in taxonomic scope, and it was therefore not clear whether the increase in rate is a general feature of endosymbiont lineages . Here, we test the prediction that a long-term reduction in Ne causes an increase in substitution rate using DNA sequences of the 16S rRNA gene from 13 phylogenetically independent comparisons between taxonomically diverse endosymbiotic microorganisms and their free-living relatives . Maximum likelihood and distance-based methods both indicate a significant increase in substitution rate in a wide range of bacterial and fungal endosymbionts compared to closely related free-living lineages . We use the same data set to test whether 16S genes from endosymbionts display increased A + T content, another indicator of increased genetic drift, and find that there is no significant difference in base composition between endosymbiont and nonendosymbiont 16S genes . However, analysis of an additional data set of whole bacterial genomes demonstrates that, while host-dependent bacteria have significantly increased genomic A + T content, the base content of the 16S gene tends to vary less than that of the whole genome . It is possible that selection for stability of rRNA is strong enough to overcome the effects of drift toward increased A + T content in endosymbiont 16S genes, despite the reduced effective population sizes of these organisms. Curr Opin Microbiol, 2003 Jun, 6(3), 310 - 6 Marker and reporter genes: illuminating tools for environmental microbiologists; Jansson JK; Fluorescent and luminescent marker and reporter genes provide easily detectable phenotypes to microbial cells and are therefore valuable tools for the study of microorganisms in the environment . Although these tools are becoming widely adopted, there are still issues that remain to be solved, such as the dependence of the reporter output on the physiological status of the cell . Eventually it might be the use of marker and reporter genes themselves that will contribute towards better understanding of the physiological status of specific microbial populations in nature. Curr Opin Microbiol, 2003 Jun, 6(3), 282 - 7 New approaches to analyzing microbial biodiversity data; Bohannan BJ et al.; Modern molecular techniques have revealed an extraordinary diversity of microorganisms, most of which are as yet uncharacterized . This poses a major challenge to microbial ecologists: how can one compare the microbial diversity of different environments when the vast majority of microbial taxa are usually unknown? Three statistical approaches developed by ecologists and evolutionary biologists--parametric estimation, nonparametric estimation and community phylogenetics--are proving to be promising tools to meet this challenge . The combination of these tools with molecular biology techniques allow the rigorous estimation and comparison of microbial diversity in different environments. Curr Opin Microbiol, 2003 Jun, 6(3), 251 - 60 Bioplastics from microorganisms; Luengo JM et al.; The term 'biomaterials' includes chemically unrelated products that are synthesised by microorganisms (or part of them) under different environmental conditions . One important family of biomaterials is bioplastics . These are polyesters that are widely distributed in nature and accumulate intracellularly in microorganisms in the form of storage granules, with physico-chemical properties resembling petrochemical plastics . These polymers are usually built from hydroxy-acyl-CoA derivatives via different metabolic pathways . Depending on their microbial origin, bioplastics differ in their monomer composition, macromolecular structure and physical properties . Most of them are biodegradable and biocompatible, which makes them extremely interesting from the biotechnological point of view. Curr Opin Microbiol, 2003 Jun, 6(3), 213 - 8 Extremophiles as a source for novel enzymes; van den Burg B; Microbial life does not seem to be limited to specific environments . During the past few decades it has become clear that microbial communities can be found in the most diverse conditions, including extremes of temperature, pressure, salinity and pH . These microorganisms, called extremophiles, produce biocatalysts that are functional under extreme conditions . Consequently, the unique properties of these biocatalysts have resulted in several novel applications of enzymes in industrial processes . At present, only a minor fraction of the microorganisms on Earth have been exploited . Novel developments in the cultivation and production of extremophiles, but also developments related to the cloning and expression of their genes in heterologous hosts, will increase the number of enzyme-driven transformations in chemical, food, pharmaceutical and other industrial applications. Curr Opin Microbiol, 2003 Jun, 6(3), 206 - 12 Microbial whole-cell sensing systems of environmental pollutants; Belkin S; The past decade has witnessed the development of a novel class of tools for environmental monitoring: genetically engineered microorganisms 'tailored' to respond in a dose-dependent manner to changes in environmental conditions . Recent advances in the field include the expansion of available reporter functions with multicolored fluorescent proteins, a broadening of the detected chemical effects such as the availability of nutrients and enhancement of the spectrum of reporter microorganisms to include cyanobacteria, yeast and fungi . Most importantly, the stage has been set for the incorporation of such cells into various whole-cell array formats on silicon chips, optic fibres and other configurations . The future of such multiplex detection and analysis systems seems bright. Nat Prod Rep, 2003 Jun, 20(3), 275 - 87 Nonribosomal peptides: from genes to products; Schwarzer D et al.; The ability to synthesize nonribosomally small bioactive peptides that find application in modern medicine is widely spread among microorganisms . As broad as the spectrum of biological activities is the structural diversity of these peptides, which are mostly cyclic or branched cyclic compounds containing non-proteinogenic amino acids, small heterocyclic rings and other unusual modifications in the peptide backbone . They are synthesized by multimodular enzymes, the so-called nonribosomal peptide synthetases (NRPSs), from simple building blocks . Biochemical and genetic studies have unveiled the key principles of nonribosomal peptide syntheses, as well as the realization of many structural features of these peptides . This review focuses on recent results in NRPS research and highlights how this knowledge can be exploited for biotechnological purposes . In addition, possibilities and limitations for prediction of structural features of uncharacterized NRPSs and approaches for their engineering are discussed. Ying Yong Sheng Tai Xue Bao, 2003 Feb, 14(2), 293 - 5 {Advances in studies on genetically engineered microorganism ecology}; Jin S et al.; Genetically engineered microorganism ecology has been one of main research contents in microorganism molecular ecology . Along with the introduction of molecular marker and molecular biology, traditional microorganism ecology has been developed; therefore, it is possible to study the relationship between GEM and environment, environmental microorganism under molecular level . The GEM ecology has become a new and intersection borderline discipline, related to molecular biology, microbiology, ecology and so on . Moreover, it brought forward that the prosecution of the research on the transgenic organism ecology and the risk assessment, and the foundation of checking means and valuating standard that adapt to the situation of China could help to the development of GEM ecology in China. Ying Yong Sheng Tai Xue Bao, 2003 Feb, 14(2), 286 - 92 {Introduction and progress of molecular microbial ecology}; Zhang H et al.; Molecular microbial ecology is an interdisciplinary field of molecular biological techniques and microbial ecology, which deals with microbial population, diversity, function, and relationships between microorganisms and biotic and abotic environments in microbial ecosystem . All the progress in molecular microbial ecology, especially in microbial diversity, phylogeny and communities of some microoganisms, indicate the renovation in traditional microbial ecology by the introduction of molecular techniques and strategies . This article reviewed the ideation, development, key domains, main methods and the future hot fields in molecular microbial ecology. Can J Diet Pract Res, 2003 Summer, 64(2), 59 - 61 Consumer food handling recommendations: is thawing of turkey a food safety issue? Lacroix BJ, Li KW, Powell DA. While it is important that dietitians and other health or food professionals provide consistent messages to the public about food safety, it is equally important that the information be evidence-based . Conflicting recommendations are evident when reviewing consumer publications from food safety advisory groups and the scientific literature . In addition, caveats are attached to the various food-handling methods . Pathogens, spoilage microorganisms, and contamination of the work area are the major concerns in thawing turkey . While several methods, including thawing on the counter at ambient temperatures, can be employed for thawing turkey, cooking to an adequate internal temperature, validated with a meat thermometer, is the more critical step . The findings indicate that providing clients or consumers with clear, consistent, evidence-based messages is difficult for food and health professionals . Further research is required to corroborate best practices in a kitchen setting . This paper is of interest to professionals who counsel clients at high risk for foodborne illness, or who counsel consumers about safe preparation of foods such as turkey. Mycol Res, 2003 Mar, 107(Pt 3), 267 - 76 Use of Coniothyrium minitans transformed with the hygromycin B resistance gene to study survival and infection of Sclerotinia sclerotiorum sclerotia in soil; Jones EE et al.; A Coniothyrium minitans strain (T3) co-transformed with the genes for beta-glucuronidase (uidA) and hygromycin phosphotransferase (hph), the latter providing resistance to the antibiotic hygromycin B, was used to investigate the survival and infection of sclerotia of Sclerotinia sclerotiorum by C . minitans over time in four different soils . Infection of sclerotia was rapid in all cases, with the behaviour of transformant T3 and wild type parent A69 being similar . Differences were seen between the soils in the rate of infection of sclerotia by C . minitans and in their indigenous fungal populations . Amendment of agar with hygromycin B enabled the quantification of C . minitan