Environ Int, 2003 Mar, 28(8), 779 - 82 Effect of heavy metals and PAH on soil assessed via dehydrogenase assay; Irha N et al.; Dehydrogenase enzyme activity (DHA) assay method using resazurin was accommodated for measuring of toxicity of compound contaminants on uncharacterized microbial communities present in any given soil . The method was used to compare the toxic effect of heavy metal and polycyclic aromatic hydrocarbon (PAH) contaminant mixture (Cr, Pb, Cu, Cd, Pyrene) on four typical Estonian soils covering a range of compositions . The method proved to be useable on all soils; the sensitivity of soil microbiology to toxic effect of contaminants was found to have a negative correlation with Ca and organic matter (OM) content and a positive correlation with amorphous phase content of soils. Pharmacogenomics, 2003 Mar, 4(2), 163 - 70 Applications of pharmacogenomics in general dental practice; Slavkin HC; The human oral cavity represents a complex ecology of approximately 500 microbial species existing as commensals, and interacting with human/host factors . Perturbation of this ecosystem can lead to diseases of the oral cavity . The oral cavity also acts as a mirror of complex systemic diseases . Unique challenges and opportunities in oral healthcare exist in the diagnosis and treatment of primary disorders of the oral cavity or manifestations of systemic diseases . The completion of human and microbial genome projects has provided a wealth of information that will permit the application of pharmacogenomics - how genetic variations within the commensal and the host will impact the efficacy of drugs, and the diagnosis and treatment of oral disease . This review highlights opportunities focusing on oral health where pharmacogenomics can be applied. Biosens Bioelectron, 2003 Apr, 18(4), 327 - 34 Operational parameters affecting the performannce of a mediator-less microbial fuel cell; Gil GC et al.; A mediator-less microbial fuel cell was optimized in terms of various operating conditions . Current generation was dependent on several factors such as pH, resistance, electrolyte used, and dissolved oxygen concentration in the cathode compartment . The highest current was generated at pH 7 . Under the operating conditions, the resistance was the rate-determining factor at over 500 omega . With resistance lower than 500 omega, proton transfer and dissolved oxygen (DO) supply limited the cathode reaction . A high strength buffer reduced the proton limitation to some extent . The DO concentration was around 6 mg l(-1) at the DO limited condition . The fact that oxygen limitation was observed at high DO concentration is believed to be due to the poor oxygen reducing activity of the electrode used, graphite . The current showed linear relationship with the fuel added at low concentration, and the electronic charge was well correlated with substrate concentration from up to 400 mg l(-1) of COD(cr) . The microbial fuel cell might be used as a biochemical oxygen demand (BOD) sensor . J Invest Dermatol, 2003 Mar, 120(3), 379 - 89 The cathelicidin anti-microbial peptide LL-37 is involved in re-epithelialization of human skin wounds and is lacking in chronic ulcer epithelium; Heilborn JD et al.; The human cathelicidin anti-microbial protein, hCAP18 is a component of the innate immune system and has broad anti-microbial activity conferred by its C-terminal fragment LL-37 . hCAP18 is constitutively produced in leukocytes and is induced in barrier organs upon inflammation and infection . We demonstrate here a novel role for this peptide in re-epithelialization of skin wounds . We show that high levels of hCAP18 are produced in skin in vivo upon wounding . The highest hCAP18 levels are attained at 48 h post-injury, declining to pre-injury levels upon wound closure . hCAP18 is detected in the inflammatory infiltrate and in the epithelium migrating over the wound bed . In chronic ulcers, however, hCAP18 levels are low and immunoreactivity for hCAP18/LL-37 is absent in ulcer edge epithelium . Using a noninflammatory ex vivo wound healing model, composed of organ-cultured human skin, we show that hCAP18 is strongly expressed in healing skin epithelium, and that treatment with antibodies raised and affinity purified against LL-37, inhibits re-epithelialization in a concentration-dependent manner . Immunoreactivity for the proliferation marker Ki67 is absent in the epithelium of such inhibited wounds, suggesting that LL-37 may play a part in epithelial cell proliferation . Thus, we suggest that, in addition to being an anti-microbial peptide, LL-37 also plays a part in wound closure and that its reduction in chronic wounds impairs re-epithelialization and may contribute to their failure to heal. Immunology, 2003 Mar, 108(3), 321 - 8 Immunomodulatory effects of cyclosporin A on human peripheral blood dendritic cell subsets; Tajima K et al.; Cyclosporin A (CsA) is a potent immuno-suppressant and is approved for the treatment of various disease conditions . The molecular biological mechanism of CsA has been investigated intensively in T cells and has been shown to involve the intracellular calcineurin pathway . Recently, it was reported that CsA has capacities to affect not only T cells but also antigen-presenting cells such as B cells and dendritic cells (DCs) . DCs are a master regulator of immune responses that have an integral capacity to prime naive T cells . In the present study, we investigated the biological effects of CsA on human peripheral blood DC subsets: CD11c+ myeloid and CD11c- lymphoid subsets . CsA inhibited the up-regulation of co-stimulatory molecules induced with or without microbial stimuli and CD40L on both CD11c+ and CD11c- subsets . In addition, CsA negatively regulated the endocytic activity of CD11c+ DC during the immature state . CsA inhibited the interleukin-12 (IL-12) production, but augmented the IL-10 production from the LPS-stimulated CD11c+ subset, whereas CsA reduced the interferon-alpha (IFN-alpha) production from the CD11c- subset infected with Sendai virus (SV) . Both the LPS-stimulated CD11c+ subset and SV-infected CD11c- subset preferentially induced the development of IFN-gamma-producing T helper-type 1 (Th1) cells . Pretreatment of these DC subsets with CsA inhibited the Th1 skewing . These findings suggested a DC-mediated mechanism of immunosuppression by CsA. Infect Control Hosp Epidemiol, 2003 Feb, 24(2), 142 - 4 Monsel's solution: a potential vector for nosocomial infection? Rupp ME, Medcalf SJ, Fey PD, Handke LD, Marion ND. Monsel's solution is a common topically applied hemostatic agent used in minor dermatologic and gynecologic surgery . Clinically, because it is often stored for long periods and dispensed from a common source for multiple patients, Monsel's solution is a potential vector for transmission of infection . However, microbiologic inoculation studies and contamination surveys indicate that Monsel's solution has properties that prohibit microbial growth, making it an unlikely vector for nosocomial infection. Ann Allergy Asthma Immunol, 2003 Feb, 90(2), 265 - 70 Hypersensitivity pneumonia-nonspecific interstitial pneumonia/fibrosis histopathologic presentation: a study in diagnosis and long-term management; Jacobs RL et al.; BACKGROUND: Nonspecific interstitial pneumonia/fibrosis (NSIP) has been classified a form of idiopathic interstitial pneumonia/fibrosis . We have shown that cases of NSIP without demonstrable serum precipitins may be caused by inhalation of high levels of mold and/or bacteria in closed environments . OBJECTIVE: We report a patient with a clinical and histopathologic diagnosis of NSIP without serum precipitins caused by a microbial contamination in her home . Her case was converted from an acute to an insidious clinical presentation by inadequate remediation . A prolonged avoidance-challenge technique demonstrated that this case of NSIP was a form of hypersensitivity pneumonia that was reversible by effective remediation . METHODS: The patient was identified by compatible signs and symptoms, roentgenographic studies, pulmonary function tests, and a transbronchial lung biopsy . She was further evaluated with a detailed environmental history, serologic tests, and investigation of the home environment . An environmental avoidance and challenge technique was performed to confirm cause and effect and to determine that remediation had been effective . RESULTS: Review of the biopsy showed NSIP and failed to reveal any non-caseating granuloma formation . Investigation of the home revealed a Cladosporium species contamination of the air conditioning system and Penicillium species beneath an entryway carpet . Serum precipitins to commercial antigens of common mold to the south Texas area were negative . Avoidance and challenge techniques confirmed the home as the causative environment in this case of NSIP . The patient has been free of signs and symptoms and has taken no medication for interstitial lung disease over the past 30 months . CONCLUSIONS: Some cases of NSIP may be caused by inhalation of microbial antigen(s) in a closed environment . An environmental challenge technique was an effective method to determine the causative environment and confirm that remediation had been effective . Inadequate remediation may lead to symptomatic improvement, but may convert a patient from an acute to an insidious presenter . The environmental challenge obviates a need for specific challenges to determine specific causation . Remediation of or moving from an environmental contamination to achieve reversibility or prevent progression was the treatment of choice to avoid use of long-term immunosuppressive agents. Phytother Res, 2003 Feb, 17(2), 97 - 106 Garlic as an antioxidant: the good, the bad and the ugly; Banerjee SK et al.; Garlic has played an important dietary and medicinal role throughout the history of mankind . In some Western countries, the sale of garlic preparations ranks with those of leading prescription drugs . The therapeutic efficacy of garlic encompasses a wide variety of ailments, including cardiovascular, cancer, hepatic and microbial infections to name but a few . However, the elucidation of its mechanism for therapeutic action has proved to be more elusive and a unifying theory, which could account for its reported multifarious activities, is yet to emerge . Reactive oxygen species (ROS) seem to be at the core of many disease processes and it is an attractive and convenient hypothesis that garlic might exert its activities through modulatory effects on ROS . A literature search on garlic and its antioxidant potential churned up a surprisingly large amount of data, some of it good, some bad and some of its definitely ugly.Various preparations of garlic, mainly aged garlic extract (AGE), have been shown to have promising antioxidant potential . However, the presence of more than one compounds in garlic, with apparently opposite biological effects, has added to the complexity of the subject . Raw garlic homogenate has been reported to exert antioxidant potential but higher doses have been shown to be toxic to the heart, liver and kidney.So where do we stand today on this issue of garlic? Is garlic always good for health? How safe is it? Is it necessary to isolate the antioxidant compounds for its medicinal use in a more effective way? These issues are addressed in this review . Soc Sci Med, 2003 Mar, 56(6), 1279 - 93 Different frames, different fears: communicating about chlorinated drinking water and cancer in the Canadian media; Driedger SM et al.; Risk issues become complicated when scientific evidence concerning a potential environmental exposure is equivocal; particularly when many argue that the public health benefits of a policy action outweigh any potential negative health effects . Chlorinated drinking water, and chlorinated disinfection byproducts (CDBPs) that are formed during the disinfection process, represent a useful case-study for examining these complications . We conduct a media analysis of chlorinated drinking water stories in the Canadian print media from 1977 to 2000 . We examine media presentations of science compared to framings by scientists, regulators, the chlorine industry, water utility representatives, and non-governmental organizations of the CDBP issue based on key informant interviews . We argue that there are two main framings of the debate, each of which are powerful in constructing risk perceptions . On the one hand, many frame the debate as a 'voluntary' risk: we choose chlorine disinfection to protect against microbial risks with a possible adverse consequence of that protection . On the other hand, others frame the issue as an 'involuntary' risk: chlorine disinfection was a 'choice' imposed by public health and water utility officials; a choice that carries a potential cancer risk, and alternative disinfection technologies are advocated . We demonstrate these different frames by examining metaphorical constructs of water, chlorine and cancer contained within them. AIDS, 2003 Mar 7, 17(4), 495 - 504 Interferon-alpha restores HIV-induced alteration of natural killer cell perforin expression in vivo; Portales P et al.; OBJECTIVE: The percentage and the activity of natural killer (NK) cells are known to be decreased in HIV-infected patients . However, the mechanisms responsible for this NK deficiency are poorly understood . Because of the role of NK cells in the host defence against microbial infections, this defect contributes to the virus-induced immune deficiency . The aim of the present study was to better understand this defect in order to be able to restore NK function in HIV infection . DESIGN AND METHODS: The expression of the cytolytic mediators perforin and granzyme A was analysed by flow cytometry, the lytic activity of peripheral blood NK cells of HIV-infected patients was analysed by cytotoxic assay, and the expression of perforin was followed during administration of interferon (IFN)alpha attached to polyethylene glycol (PEG)-IFNalpha . RESULTS: The lytic activity and the expression of perforin and granzyme A was low in NK cells of infected individuals in comparison with normal control volunteers . In both groups NK cytotoxic capacity was linked to perforin expression . The low perforin expression in HIV-infected subjects negatively correlated with HIV RNA plasma level . administration of PEG-IFNalpha restored perforin expression even in patients whose viral load was not reduced by this treatment . CONCLUSIONS: These results suggest that HIV-induced NK deficiency could be partly mediated by a defect in perforin and granzyme A expression, and that PEG-IFNalpha could be used in infected subjects to directly improve their natural immunity in addition to eventually reducing their viraemia. Microsc Microanal, 2001 Nov, 7(6), 470 - 485 In Situ Environmental Cell-Transmission Electron Microscopy Study of Microbial Reduction of Chromium(VI) Using Electron Energy Loss Spectroscopy; Daulton TL et al.; Reduction of Cr(VI) by the bacterium, Shewanella oneidensis (previously classified Shewanella putrefaciens strain MR-1), was studied by absorption spectrophotometry and in situ, environmental cell-transmission electron microscopy (EC-TEM) coupled with electron energy loss spectroscopy (EELS) . Bacteria from rinsed cultures were placed directly in the environmental cell of the transmission electron microscope and examined under 100 Torr pressure . Bright field EC-TEM images show two distinct populations of S . oneidensis in incubated cultures containing Cr(VI)O2- 4: those that exhibit low image contrast and heavily precipitate-encrusted cells exhibiting high image contrast . Several EELS techniques were applied to determine the oxidation state of Cr associated with encrusted cells . The encrusted cells are shown to contain a reduced form of Cr in oxidation state +3 or lower . These results demonstrate the capability to determine the chemistry and valence state of reduction products associated with unfixed, hydrated bacteria in an environmental cell transmission electron microscope. Crit Care Nurs Clin North Am, 2003 Mar, 15(1), 89 - 96, ix Economics of antibiotic administration; Sommers BD; This article examines several elements of antibiotic administration that make it worthy of policy analysis, including microbial resistance, contagion, competing brand and generic drugs, and formulary restrictions by insurers and hospitals . These topics are explored using two concepts from health economics, cost-effectiveness and externalities, revealing theoretical and empirical evidence that society may not be using antibiotics as efficiently as it could. Life Sci Space Res, 1977, 15, 59 - 63 Determinations of microbial loads associated with microscopic-size particles of Kennedy Space Center soil; Ruschmeyer OR et al.; Plate counts for six fractions, of micrometer-size, of Kennedy Space Center soil provided estimates of aerobic, mesophilic, heterotrophic, microbial loads on single soil particles . Analyses included unheated particles, particles subjected to wet heat at 80 degrees C for 20 min, and particles subjected to dry heat at 110 degrees C for 1 hr . Unheated particles yielded mean counts ranging from 6 colonies per particle for the smallest (44-53 micrometers) soil fraction to approximately 55 colonies per particle for the largest size (105-125 micrometers) soil fraction tested . Mean counts for heat-resistant forms ranged from 2 colonies per particle for the smaller particles to 12-15 colonies for the largest particles analyzed. Neurochirurgie, 2002 Dec, 48(6), 516 - 21 {Nocardia farcinica brain abscess: clinical and specific radiological findings and management . Report of two cases in immunononcompromised patients}; Vialle R et al.; Nocardia spp . cerebral abscesses are rare and usually occur in immunocompromised patients . We report two recent cases of cerebral abscesses due to Nocardia farcinica in immunocompetent patients and review the literature about diagnosis and therapeutic issues . Outcome was good for the two patients following an early identification of the bacteria . Stereotactic biopsy was performed in one case and craniotomy with excision of the abscess in the other case . In both cases, complete identification of the bacteria could be achieved, followed by prolonged antibiotic therapy . Exposure to the germ (mainly telluric) is often difficult to suspect from the past medical history of the patient . Diagnosis is also unexpected . Despite the typically and characteristic aspect on CT and MRI, specific identification and anti-microbial sensitivity profiles are necessary to optimize treatment . In some rare cases, unusual species like Nocardia farcinica, can be resistant to numerous antibiotics requiring adjustments of medical management . Early identification of the bacteria is necessary to achieve good outcome in immunocompetents patients. Philos Trans R Soc Lond B Biol Sci, 2003 Jan 29, 358(1429), 191 - 201; discussion 201-2 Mitochondria and hydrogenosomes are two forms of the same fundamental organelle; Embley TM et al.; Published data suggest that hydrogenosomes, organelles found in diverse anaerobic eukaryotes that make energy and hydrogen, were once mitochondria . As hydrogenosomes generally lack a genome, the conversion is probably one way . The sources of the key hydrogenosomal enzymes, pyruvate : ferredoxin oxidoreductase (PFO) and hydrogenase, are not resolved by current phylogenetic analyses, but it is likely that both were present at an early stage of eukaryotic evolution . Once thought to be restricted to a few unusual anaerobic eukaryotes, the proteins are intimately integrated into the fabric of diverse eukaryotic cells, where they are targeted to different cell compartments, and not just hydrogenosomes . There is no evidence supporting the view that PFO and hydrogenase originated from the mitochondrial endosymbiont, as posited by the hydrogen hypothesis for eukaryogenesis . Other organelles derived from mitochondria have now been described in anaerobic and parasitic microbial eukaryotes, including species that were once thought to have diverged before the mitochondrial symbiosis . It thus seems possible that all eukaryotes may eventually be shown to contain an organelle of mitochondrial ancestry, to which different types of biochemistry can be targeted . It remains to be seen if, despite their obvious differences, this family of organelles shares a common function of importance for the eukaryotic cell, other than energy production, that might provide the underlying selection pressure for organelle retention. Eur J Immunol, 2003 Jan, 33(1), 101 - 7 The ability of murine dendritic cell subsets to direct T helper cell differentiation is dependent on microbial signals; Manickasingham SP et al.; Dendritic cells (DC) initiate T cell responses and direct the class of T cell immunity through the production of Th-polarizing cytokines . In the mouse, immunization with CD8alpha(+) DC has led to Th1 priming whereas immunization with CD8alpha(-) DC has been associated with Th2 induction . Here, we use a direct T cell priming assay in vitro to re-examine the Th-directing potential of total DC or purified CD4(+) DC, CD8alpha(+) DC or CD4(-) CD8alpha(-) (double-negative; DN) DC subsets from mouse spleen . We show that the default Th effector phenotype induced by priming with DC depends on the protocol used for T cell purification, the T cell:antigen-presenting cell ratio and the antigen dose but is only marginally affected by DC subtype . All DC subsets can direct increased Th1 development in response to microbial stimuli known to elicit IL-12 production . Similarly, all subsets can suppress Th1 development and allow Th2 cellsto expand upon exposure to IL-10-inducing microbial agents . The flexibility of DC in directing Th development in function of microbial signals argues against the notion of pre-determined "DC1" and "DC2" subsets and suggests that multiple DC subtypes can direct an appropriate Th response to different classes of infectious agents. J Immunol, 2003 Mar 1, 170(5), 2409 - 16 The 19-kDa Mycobacterium tuberculosis protein induces macrophage apoptosis through Toll-like receptor-2; Lopez M et al.; Macrophages infected with Mycobacterium tuberculosis undergo increased rates of apoptosis . Important objectives are to define the microbial factors that cause apoptosis, the mechanisms involved and the impact on infection . The 19-kDa M . tuberculosis glycolipoprotein (p19) is both cell wall-associated and secreted and is a candidate virulence factor . We investigated the potential of recombinant, His-tagged p19 lacking the secretion/acylation signal to induce macrophage apoptosis . The TUNEL assay and annexin V binding to membrane phosphatidylserine were used to measure apoptosis . The results show that p19 does act to induce apoptosis in differentiated THP-1 cells and monocyte-derived macrophages and that this effect is both dose- and time-dependent . Furthermore, this effect of p19 is Toll-like receptor (TLR)-2-mediated because preincubation of either THP-1 cells or TLR-2-expressing CHO cells with anti-TLR-2 mAb inhibited apoptosis induced by p19 . Apoptosis of macrophages in response to p19 was found to be caspase-8 dependent and caspase-9 independent consistent with a transmembrane pathway signaling cell death through TLR-2 . The viability of M . tuberculosis in cells undergoing apoptosis induced by p19 was significantly reduced suggesting the possibility that this may favor containment of infection . Although native p19 is a mycobacterial glycolipoprotein, based upon the use of recombinant p19 where the acylation signal had been removed, we conclude that it is the polypeptide component of p19 that is responsible for signaling through TLR-2 and that the lipid moiety is not required. J Periodontol, 2003 Jan, 74(1), 90 - 6 Molecular interaction of Porphyromonas gingivalis with host cells: implication for the microbial pathogenesis of periodontal disease; Amano A; Porphyromonas gingivalis is a predominant periodontal pathogen, which expresses a number of potential virulence factors involved in the pathogenesis of periodontitis . Among them, fimbriae are a critical factor to mediate the bacterial interaction with host tissues, which promotes the bacterial adhesion to and invasion of the targeted sites . Fimbriae are capable of binding to human salivary components, commensal bacteria, and a variety of host cells including macrophages, epithelial cells, and fibroblasts . Human extracellular matrix (ECM) proteins such as vitronectin and fibronectin play important roles in cellular signal transduction via binding to receptor integrins . Fimbriae showed significant binding affinity to ECM proteins and clearly inhibited the molecular interactions between vitronectin/fibronectin and their receptor alphavbeta3 and alpha5beta1 integrins overexpressed on Chinese hamster ovary (CHO) cell strain . P . gingivalis fimbriae are likely to interrupt the cellular signaling via ECM proteins/integrins in periodontal regions . Fimbriae are also thought to be critically important in invasive events of the organism to host cells . The fimA genes, encoding FimA (a subunit of fimbriae), of P . gingivalis strains are classified into 5 types, I to V . Recent clinical investigations demonstrated the close relationship between the organisms with type II fimA and periodontitis development . Recombinant FimA (rFimA) proteins of types I to V were generated to compare their adhesion/invasion abilities to human gingival fibroblasts (HGF) and a human epithelial cell line (HEp-2 cells), respectively . There were no significant differences in the adhesion ability of microspheres (MS) coated with these rFimAs to HGF; however, the adhesion of type II rFimA-MS to HEp-2 cells was significantly greater than that of other rFimA types . It was also observed that the type II rFimA-MS markedly invaded the epithelial cells and accumulated around the nuclei . Collectively, these findings suggest that fimbriae of P . gingivalis, especially type II, are involved in the initiation and progression of human periodontitis. DNA Seq, 2002 Oct, 13(5), 301 - 6 Cloning and characterization of a novel human alcohol dehydrogenase gene (ADHFe1); Deng Y et al.; There are three types of NAD(P)(+)-dependent alcohol dehydrogenase: "medium-chain" zinc-containing enzymes, "short-chain" zinc-lacking enzymes and iron-activated enzymes . Although the medium-chain family and the short-chain family have been characterized in human, the iron-activated alcohol dehydrogenase family seems to exist only in microbial organisms . We have now cloned and characterized an iron-activated alcohol dehydrogenase gene, Fe-containing alcohol dehydrogenase 1 (ADHFe1) in human . The cDNA was isolated from the human fetal brain cDNA library . It contains a long open reading frame, which is not homologous with the other alcohol dehydrogenases isoenzymes characterized in human . The hypothetical alcohol dehydrogenase does, however, show strong homology to the iron-activated alcohol dehydrogenases from microbial organisms . Northern blotting analysis only detected expression in adult liver tissue . At least two different splicing variants were screened by PCR using the multiple tissue cDNA panels as templates. Dermatology, 2003, 206(2), 96 - 105 The role of innate immunity in the pathogenesis of acne; Koreck A et al.; Acne is a multifactorial disease of the pilosebaceous follicle . The most significant pathogenetic factors of acne are: abnormal ductal keratinization, increased sebum secretion, abnormalities of the microbial flora and inflammation . The pilosebaceous unit is an immunocompetent organ . Keratinocytes and sebocytes may act as immune cells capable of pathogen recognition and abnormal lipid presentation, and they might have an important role in initiating and perpetuating the activation of both innate and adaptive immune responses . The elements of the skin immune system are involved in the development of both noninflammatory and inflammatory acne lesions . Phytochemistry, 2003 Apr, 62(7), 1101 - 4 Microbial transformation of zaluzanin-D; Kumari GN et al.; Microbial transformation of zaluzanin-D using different fungi gave 11,13-dihydrozaluzanin-C, zaluzanin-C, 4,16,11,13 - tetrahydro zaluzanin-C, estafiatone, dihydroestafiatol and dihydroestafiatone. J Agric Food Chem, 2003 Feb 26, 51(5), 1460 - 3 Factors Influencing texture retention of salt-free, acidified, red bell peppers during storage; Papageorge LM et al.; Red bell peppers were stored in a salt-free, acidified cover solution with sulfite as a microbial preservative . The texture retention of stored peppers was evaluated as a function of pH, acid, calcium, blanch treatment, and growing conditions of the peppers . Field-grown peppers softened at a faster rate than greenhouse-grown peppers and exhibited more variability in their ability to maintain texture during storage . Improved firmness retention of red peppers stored at 30 degrees C was observed when the pH was adjusted to be in the range of 3.4-3.8, when at least 5 mM calcium was added to the peppers, and when red peppers were blanched at 75 degrees C for at least 1 min. J Agric Food Chem, 2003 Feb 26, 51(5), 1347 - 52 Fipronil metabolism and dissipation in a simplified aquatic ecosystem; Aajoud A et al.; Several phenylpyrazole derivatives are selective inhibitors of chloride channel activities in insects . In this chemical family, fipronil is a powerful insecticide now widely used for several purposes . The dissipation of this molecule in a simplified aquatic ecosystem has been studied for 3 months, using (14)C-labeled fipronil . The main features of the complex process leading to fipronil transformation in this system were the following . The fipronil aqueous solution was submitted to two chemical transformations: the photodependent desulfuration of the side chain bound to the 4-position of the heterocyclic ring and the chemical hydrolysis of the nitrile function bound to the 3-position . Fipronil, rapidly transferred from the water solution to the organic matter, was protected from the previously mentioned chemical transformations but evolved to give two main metabolites, which were either reduced or oxidized in the side chain on the 4-position . These derivatives were powerful insecticides as shown by LC(50) measurements on Aedes aegypti larvae (LC(50) for CF(3)-S-R and CF(3)-SO(2)-R = 8.8 nM) . During the course of this experiment, nitrile hydrolysis took place slowly, originating either from the chemical hydrolysis in the aqueous solution or from enzymatic hydrolysis inside the microbial biomass . The fipronil-amide (3-NH(2)-CO-R') derivative, although much more polar than fipronil itself, was mostly bound to the organic matter . Other more polar derivatives were also detected but in very small amounts . No (14)CO emission was observed during the experiment. Clin Chim Acta, 2003 Mar, 329(1-2), 95 - 102 A new method to evaluate spontaneous platelet aggregation in type 2 diabetes by Cellfacts; Gabbianelli R et al.; BACKGROUND: The alterations in the functional activities of platelets in diabetes produce an increase of spontaneous platelet aggregation (SPA) and release of platelet-derived microparticles . Platelet-derived microparticles are shed from platelets during activation by high shear stress, collagen and certain agonists . Although the physiologic role of microparticles has been difficult to assess, the characterization of their biological activity is of interest in view of a possible role in hemostasis and coagulation and their reported involvement in thrombotic disease . METHODS: We propose a new, simple method to evaluate spontaneous platelet aggregation and release of platelet-derived microparticles by the Cellfacts analyser (Microbial System Limited (MSL), Coventry, England) that uses electrical sensing flow impedance determination to detect the size particles and the cells in a conductive fluid . Platelet-rich plasma (PRP) from type 2 diabetes was employed for this study . The importance of platelet-activating factor (PAF) on spontaneous platelet aggregation was evaluated and the effect of vitamin E and WEB 2086-BS, an antagonist of platelet-activating factor, was measured . RESULTS AND CONCLUSIONS: Data presented show that Cellfacts could be an easy and fast instrument to check the state of platelets in patients with alterations in the functionality of platelets, and to follow the effect of pharmacological therapy on spontaneous platelet aggregation and the release of platelet-derived microparticles. Biochem Biophys Res Commun, 2003 Feb 21, 301(4), 1062 - 8 Inhibition of Epstein-Barr virus lytic cycle by (-)-epigallocatechin gallate; Chang LK et al.; (-)-Epigallocatechin gallate (EGCG), abundant in green tea, is a potent anti-microbial and anti-tumor compound . This investigation used immunoblot, flow cytometry, microarray, and indirect immunofluorescence analyses to show that at concentrations exceeding 50 microM, EGCG inhibits the expression of Epstein-Barr virus (EBV) lytic proteins, including Rta, Zta, and EA-D, but does not affect the expression of EBNA-1 . Moreover, DNA microarray and transient transfection analyses demonstrated that EGCG blocks EBV lytic cycle by inhibiting the transcription of immediate-early genes, thus inhibiting the initiation of EBV lytic cascade. Oral Microbiol Immunol, 2003 Feb, 18(1), 59 - 65 Denaturing gradient gel electrophoresis analysis to study bacterial community structure in pockets of periodontitis patients; Zijnge V et al.; Bacteria are involved in the onset and progression of periodontitis . A promising molecular technique, denaturing gradient gel electrophoresis (DGGE), to study microbial population dynamics in the subgingival pocket is presented . Twenty-three samples were taken from the subgingival pockets of nine patients and six healthy family members . From four periodontitis patients, 12 samples were evaluated before, 1 day after and 3 months after treatment . Part of the 16S rRNA gene of all bacteria was amplified by PCR and separated by DGGE, creating banding patterns representative of the community structure . Shifts in composition and diversity of the microbial population could be determined semiquantitatively, and this showed that treatment resulted in a decrease in the diversity of the population . After 3 months a microbial population 33-47% different from the population before treatment had re-established . Intense bands representing Exiguobacterium aurantiacum were present in 13 out of 25 samples, indicating that this species may play a role in periodontal disease. FEBS Lett, 2003 Feb 11, 536(1-3), 77 - 84 Unexpected catalytic site variation in phosphoprotein phosphatase homologues of cofactor-dependent phosphoglycerate mutase; Rigden DJ; The cofactor-dependent phosphoglycerate mutase (dPGM) superfamily contains, besides mutases, a variety of phosphatases, both broadly and narrowly substrate-specific . Distant dPGM homologues, conspicuously abundant in microbial genomes, represent a challenge for functional annotation based on sequence comparison alone . Here we carry out sequence analysis and molecular modelling of two families of bacterial dPGM homologues, one the SixA phosphoprotein phosphatases, the other containing various proteins of no known molecular function . The models show how SixA proteins have adapted to phosphoprotein substrate and suggest that the second family may also encode phosphoprotein phosphatases . Unexpected variation in catalytic and substrate-binding residues is observed in the models. Syst Appl Microbiol, 2002 Dec, 25(4), 555 - 71 Improvement of ciliate identification and quantification: a new protocol for fluorescence in situ hybridization (FISH) in combination with silver stain techniques; Fried J et al.; A new protocol for taxon specific probe based fluorescent in situ hybridization was developed for the identification and quantification of ciliates in microbial communities . Various fixatives and experimental parameters were evaluated and optimized with respect to cell permeability and morphological preservation . Optimum results were adaption by obatined of a modified fixation method using Bouin's solution . Furthermore, conventional staining procedures such as different Protargol stain techniques and a silver nitrate impregnation method were modified and can now be applied in combination with fluorescence in situ hybridization . The new protocol allows a rapid and reliable identification as well as quantification of ciliates based upon classical morphological aspects and rRNA based phylogenetic relationships performed in one experiment . Furthermore, a set of specific probes targeting different regions of the 18S rRNA was designed for Glaucoma scintillans Ehrenberg, 1830 and tested by applying this new approach of combining in situ cell hybridization with conventional staining techniques. Chirality, 2003 Mar, 15(3), 220 - 6 Lipase regio- and stereoselectivities toward three enantiomeric pairs of didecanoyl-deoxyamino-O methyl glycerol: a kinetic study by the monomolecular film technique; Douchet I et al.; A kinetic study was carried out on the regio- and stereoselectivities of 12 lipases of animal and microbial origin . For this purpose, monomolecular films consisting of three pairs of enantiomers (didecanoyl-deoxyamino-O methyl glycerol, DDG) containing a single hydrolyzable decanoyl ester bond and two lipase-resistant groups were spread at the air-water interface . Each of the lipases tested displayed a particular type of behavior, on the basis of which they were classified in two groups, depending on their ability to hydrolyze the sn-2 position . From the qualitative point of view, the sn-2 preference measured on triacylglycerides and DDG were in good agreement . The inductive chemical effect might explain why a greater level of hydrolytic activity was observed with the diglycerides than with DDG . With most of the lipases tested, it was observed that the enantiomeric pair having two distal acyl chains was more clearly differentiated stereochemically than the two homologous pairs with two adjacent acyl chains . This finding is consistent with the hypothesis that during the chiral recognition process two of the three attachment points may be the external (distal) hydrophobic chains, which is in line with the hypothesis of a tuning fork conformation of a triglyceride in the lipase active site . AIDS Res Hum Retroviruses, 2003 Jan 1, 19(1), 21 - 9 HIV-mediated gammadelta T cell depletion is specific for Vgamma2+ cells expressing the Jgamma1.2 segment; Enders PJ et al.; Circulating Vgamma2/Vdelta2(+) T cells, normally constituting 3-6% of all CD3(+) T cells in blood, are severely depleted after HIV infection . The mechanism(s) for Vgamma2/Vdelta2(+) T cell depletion are unknown, partly because these cells are CD4(-) and resistant to HIV infection . To determine whether this cell depletion was general for all Vgamma2(+) cells or specific for an individual subset, we analyzed the Vgamma2 repertoire and found consistent differences between HIV(+) and uninfected control samples . The change in Vgamma2 repertoire was the result of preferentially depleting only those Vgamma2 cells that express the Jgamma1.2 segment . The specific loss of Vgamma2-Jgamma1.2(+) cells was polyclonal, as the Vgamma subset retained normal diversity even after HIV infection, and loss occurred without significant changes in the paired chain (Vdelta2) repertoire, or in the alternate Vdelta1 chain repertoire . Specific depletion of Vgamma2-Jgamma1.2/Vdelta2 T cells is the first evidence of a common, T cell receptor-dependent cell loss in HIV disease and it provides a clear example of bystander cell depletion . Vgamma2-Jgamma1.2/Vdelta2 T cells mediate potent responses to microbial pathogens including HIV, and loss of this subset is an important aspect of AIDS pathogenesis. Lett Appl Microbiol, 2003, 36(3), 140 - 4 Pulsed electric fields cause sublethal injury in Escherichia coli; Garcia D et al.; AIMS: The objective was to investigate the occurrence of sublethal injury in Escherichia coli by pulsed electric fields (PEF) at different pH values . METHODS AND RESULTS: The occurrence of sublethal injury in PEF-treated E . coli cells depended on the pH of the treatment medium . Whereas a slight sublethal injury was detected at pH 7, 99.95% of survivors were injured when cells were treated at pH 4 for 400 micros at 19 kV . The PEF-injured cells were progressively inactivated by a subsequent holding at pH 4 . CONCLUSIONS: PEF cause sublethal injury in E . coli . The measurement of sublethal injury using a selective medium plating technique allowed prediction of the number of cells that would be inactivated by subsequent storage in acidic conditions . SIGNIFICANCE AND IMPACT OF THE STUDY: This work could be useful for improving food preservation by PEF technology and contributes to the knowledge of the mechanism of microbial inactivation by PEF. Poult Sci, 2003 Jan, 82(1), 92 - 9 Effects of phytase supplementation on the performance, egg quality, and phosphorous excretion of laying hens fed different levels of dietary calcium and nonphytate phosphorous; Lim HS et al.; An experiment employing a factorial arrangement of two levels (3.0 and 4.0%) of Ca, two levels (0.15 and 0.25%) of nonphytate phosphorus (NPP), and two levels (0 and 300 U/kg diet) of microbial phytase was carried out with 960 ISA-brown layers from 21 to 41 wk of age . There was a significant interaction between NPP level and phytase for egg production . High NPP level and phytase supplementation increased egg production only in the second 10-wk period (31 to 41 wk) . High NPP and low Ca increased feed intake, and a significant interaction between levels of NPP and Ca was observed in the first 10 wk . High NPP improved feed efficiency only in the second 10-wk period . Low NPP improved egg specific gravity and eggshell thickness but decreased Haugh units in the first 10-wk period; high NPP decreased the percentage of broken and soft-shell eggs in the second period . Low Ca decreased egg specific gravity, eggshell strength, and eggshell thickness in both periods and increased Haugh units in the second 10-wk period . Phytase supplementation decreased the percentage of broken and soft-shell eggs . High NPP increased fiber availability but decreased Ca availability . High Ca decreased Ca availability, whereas phytase increased availability of dry matter, fiber, and P . High NPP increased retention of P and Fe but also increased excretion of P . High Ca decreased retention of Zn and Fe . Phytase supplementation increased P retention, resulting in decrease of P excretion . In conclusion, supplementation of microbial phytase at a level of 300 U per kg diet of laying hens can improve egg production, decrease broken and soft egg production rate, and P excretion . The effects of phytase supplementation are significantly modified by the level of Ca and NPP. Int J Mol Med, 2003 Mar, 11(3), 395 - 400 Stem cell transplantation for treatment of liver diseases: from biological foundations to clinical experience (review); Sukhikh GT et al.; The insufficiency of liver functions remains one of the major causes of death in different regions of the world . This syndrome is an ultimate outcome of irreversible damage of liver parenchyma by a variety of stimuli such as environmental toxins and microbial metabolites . The acute and/or chronic inflammation caused by these agents leads to massive apoptosis of hepatocytes, highly specialized polyfunctional epithelial cells . A decrease in the number of hepatocytes below the critical level dictates the necessity of supplanting the exogenous viable cells . Such cells must fulfill two major requirements: i) they should be capable of proliferating in the recipient's liver, and ii) they should differentiate into mature (i.e, functionally competent) hepatocytes . Evidence is growing in support of the role of early pluripotent precursors (stem cells of various origin) as a unique source for transplantation . This review analyses the state-of-the-art in the fields of developmental biology, cytology, molecular physiology and biochemistry of fetal stem cells with the emphasis on their use in experimental models of liver insufficiency . These biological considerations validate the therapeutic manipulations with fetal cells in clinical trials. Zhongguo Shi Yan Xue Ye Xue Za Zhi, 2001 Jun, 9(2), 153 - 159 {Operating Procedure of Collection, Processing and Preservation of 3000 Units Umbilical Cord Blood in Shangdong Cord Blood Bank}; Zhou SL et al.; The experience with the umbilical cord blood (UCB) stem cells for unrelated transplantation from our 3 000 UCB storage was described . UCB, collected from closed blood bags, were mixed with hydroxyethyl starch for nucleated cell (NC) enrichment . After finishing CD34 analysis, culture of hematopoietic progenitors (CFU-GM and CFU-GEMM) assays, microbial culture, HLA Class I (A, B) serology and class II (DR) low resolution SSP typing, cord blood units are stored in the liquid nitrogen for clinical applicatoin . Cord blood contained an average of nuclear cell (NC) (1.2 +/- 0.6) x 10(9), CD34(+) cells (3.0 +/- 3.7) x 10(6), CFU-GM (1.1 +/- 0.7) x 10(6) and CFU-GEMM (1.1 +/- 1.2) x 10(6) for storage and the recovery rates were 91%, 88%, 85% and 82%, respectively . The recovery rates for red blood cell and Hb were (39 +/- 9)% and (40 +/- 8)%, respectively . The storage volume was (35.1 +/- 7.1) ml in a 50 ml storage bags . The mean time from collection to processing of 15 hours (range 4 - 24 hours) had no influence on cell viability . The cell viability before processing is more than 95% and 92% after UCB thawing . The recovery rates of NC, CD34(+) cells and CFU-GM post-thawing were 96%, 90% and 91%, respectively . There were no HIV antibody (HIVAb) positive in all of UCB units . For an incidence of processed samples, infection with syphilis, HBsAg, HBcAb, HCVAb, CMV, bacterial contamination and abnormal hemoglobin were 0.1%, 0.8%, 3.2%, 0.2%, 87.1%, 1.2% and 0.1%, respectively . More than 3 HLA loci matched can be found for random patients in our cord blood bank and 6 HLA loci matched have 5% . For transplantation with nucleated cell counts of > 2.7 x 10(7) cells/kg, our cord blood bank will be able to provide all of the umbilical cord blood stem cell samples for children and 50% of units can be used for some of adult recipients transplantation in the country . It is concluded that: (1) The large cord blood banking for 20 000 UCB storage is feasible in China . (2) Our system of whole procedure and methods is functionable for supplying qualified cord blood units in transplantation . (3) The volume for collection is critical to the yield of CD34(+) cells or hematopoietic progenitor cells, however cord blood NC is also important and proportional with CD34(+) cells . Only the units containing more than 8 x 10(8) cells and more than 60 ml of cord blood can be in the procession for storage. J Heart Valve Dis, 2003 Jan, 12(1), 68 - 75 Persistence of Chlamydia pneumoniae in degenerative aortic valve stenosis indicated by heat shock protein 60 homologues; Skowasch D et al.; BACKGROUND AND AIMS OF THE STUDY: Based on the concept of chronic persistent infections with Chlamydia pneumoniae among variable stressors for aortic valve degeneration, the study aim was to assess the presence of chlamydial heat shock protein (cHSP) 60 and its human homologue (hHSP60) in diseased valvular tissue . METHODS: Surgical specimens of high-grade stenosed, native (n = 33) and bioprosthetic (n = 10) aortic valves were examined immunohistochemically for the localization of cHSP60, hHSP60 and macrophages (CD68), supplemented by polymerase chain reaction (PCR) and electron microscopy to prove microbial presence . RESULTS: Degenerated valves showed specific immunostaining of cHSP60 in 27 cases (65%), of hHSP60 in 26 (63%), and of CD68 in 36 (84%) . Both HSP60 homologues were predominantly detected in valvular fibrosa, consistently co-localized with macrophages and, quantitatively, showed a strong correlation (r = 0.81, p < 0.001) . Presence of C . pneumoniae was demonstrated by PCR in a subset of 11 of 18 valves (61%) . Microbial persistence was confirmed by ultrastructural analysis . Degenerated prosthetic valves revealed markedly higher macrophage infiltration and cHSP60 signaling compared with degenerated native valves (each p < 0.05) . CONCLUSION: Beyond detection of C . pneumoniae, the present data on co-localization and valvular predilection sites (fibrosa) of both HSP60 homologues indicate the presence of chronic persistent C . pneumoniae infection as well as regional stressor effects, and suggest their involvement in native and prosthetic valve degeneration. J Lipid Res, 2003 Feb, 44(2), 342 - 8 Epub 2002 Nov 04. Effect of GM2 activator protein on the enzymatic hydrolysis of phospholipids and sphingomyelin; Shimada Y et al.; GM2 activator protein (GM2AP) is a specific protein cofactor that stimulates the enzymatic hydrolysis of the GalNAc from GM2, a sialic acid containing glycosphingolipid, both in vitro and in lysosomes . While phospholipids together with glycosphingolipids are important membrane constituents, little is known about the possible effect of GM2AP on the hydrolysis of phospholipids . Several recent reports suggest that GM2AP might have functions other than stimulating the conversion of GM2 into GM3 by beta-hexosaminidase A, such as inhibiting the activity of platelet activating factor and enhancing the degradation of phosphatidylcholine by phospholipase D (PLD) . We therefore examined the effect of GM2AP on the in vitro hydrolyses of a number of phospholipids and sphingomyelin by microbial (Streptomyces chromofuscus) and plant (cabbage) PLD . GM2AP, at the concentration as low as 1.08 microM (1 microg/50 microl) was found to inhibit about 70% of the hydrolyses of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol by PLD, whereas the same concentration of GM2AP only inhibited about 20-25% of the hydrolysis of sphingomyelin by sphingomyelinase and had no effect on the hydrolysis of sphingosylphosphorylcholine by PLD . Thus, GM2AP exerts strong and broad inhibitory effects on the hydrolysis of phospholipids carried out by plant and microbial PLDs . High ammonium sulfate concentration (1.6 M or 21.1%) masks this inhibitory effect, possibly due to the alteration of the ionic property of GM2AP. Int J Immunopathol Pharmacol, 2002 Sep, 15(3), 245 - 248 Prevalence of IgG antibodies against Borrelia Burgdorferi s.l . and Ehrlichia Phagocytophila in sera of patients presenting symptoms of Lyme disease in a central region of Italy; Santino I et al.; The aim of this study was to evaluate the prevalence (seroprevalence) of antibodies against Borrelia burgdorferi and Ehrlichia phagocytophila among patients resident in Lazio, a region of central Italy . Of a sample of 1,050 patients, which presented clinical manifestations related to Lyme disease, 34 (3.2%) were Borrelia-seropositive (Lyme index value >/= 1.2) . The sera of 25 out of the 34 patients that were Borrelia-positive were also analysed for the presence of antibodies against E . phagocytophila and 3 (12%) were found Ehrlichia-positive (titres >1:64) . No Ehrlichia positive samples were found among sera of 250 Borrelia-negative patients . Since both B . burgdorferi s.l . and Ehrlichia species share the same tick vector (Ixodes ricinus), our results indicate that concurrent transmission of these microbial pathogens might have been occurred among the patients included in this study. J Immunol, 2003 Feb 15, 170(4), 1763 - 9 Expression of IFN-gamma upon triggering of activating Ly49D NK receptors in vitro and in vivo: costimulation with IL-12 or IL-18 overrides inhibitory receptors; Ortaldo JR et al.; NK cells can express both activating and inhibitory Ly49 receptors on their cell surface . When cells expressing both receptors are presented with a ligand, inhibition dominates the functional outcome . In this report we demonstrate that costimulation of the activating Ly49D murine NK cell receptor with IL-12 or IL-18 is capable of over-riding the inhibitory Ly49G2 receptor blockade for cytokine production both in vitro and in vivo . This synergy is mediated by and dependent upon Ly49D-expressing NK cells and results in significant systemic expression of IFN-gamma . This would place NK cells and their activating Ly-49 receptors as important initiators of microbial, antiviral, and antitumor immunity and provide a mechanism for the release of activating Ly49 receptors from inhibitory receptor blockade. Appl Occup Environ Hyg, 2003 Mar, 18(3), 193 - 9 Extent of fungal growth on fiberglass duct liners with and without biocides under challenging environmental conditions; Samimi BS et al.; Eight brands of fiberglass duct liners, including three that contained biocides, were exposed to challenging environmental conditions that would promote fungal growth . Twenty-four rectangular sheet metal ducts in three groups of eight ducts per group were lined with the eight selected liners . Each group of ducts was exposed to one of the three test conditions within an environmental chamber for a period of 15 days . These conditions were a) 75 percent RH, b) 75 percent RH plus water spray, c) 75 percent RH plus dry nutrient, and d) 75 percent RH plus water plus nutrient . Viable spores of Aspergillus niger were aerosolized into each duct as seed . On the 16th day, air and surface samples for fungal spores were collected from inside ducts . The results of air sampling using N6 sampler and visual inspection indicated that two out of three biocide-containing liners, Permacote and Toughgard, inhibited fungal growth but only under condition A . The third biocide-containing liner, Aeroflex Plus, was effective even when it was wet (conditions A and B) . All three biocide-containing liners failed to inhibit fungal growth under conditions C and D . Among the five other types of liners that did not contain biocides, ATCO Flex with a smooth Mylar coating was more preferable, exhibiting lower fungal activity during conditions A, B, and C . All liners failed under condition D when nutrient and water were added together . Surface sampling using adhesive tape failed to produce representative results, apparently due to rough/porous surface of duct liners . It was concluded that duct liners with biocide treatment could be less promoting to microbial growth under high humidity as long as their surfaces remain clean and water-free . A liner with an impermeable and smooth surface seems to be less subject to microbial growth under most conditions than biocide-containing liners having porous and/or rough surfaces. Biometals, 2003 Jun, 16(2), 359 - 68 Early onset of virus infection and up-regulation of cytokines in mice treated with cadmium and manganese; Seth P et al.; A substantial database indicates that a large number of environmental pollutants, chemicals and therapeutic agents to which organisms are exposed cause immunotoxicity . The suppression of immune functions may cause increased susceptibility of the host to a variety of microbial pathogens potentially resulting in a life-threatening state . Evaluation of the immunotoxic potential of chemical xenobiotics is of great concern and, therefore, we have investigated the impact of exposure of inorganic metals, specifically cadmium (Cd) and manganese (Mn) on Encephalomyocarditis virus (EMCV), Semliki Forest virus (SFV), and Venezuelan Equine Encephalitis virus (VEEV) infection . Pretreatment with a single, oral dose of Cd or Mn increased the susceptibility of mice to a sub-lethal infection of these viruses as observed by increased severity of symptoms and mortality compared to untreated controls . An early onset of virus infection was found in brains of Cd and Mn treated animals . Histopathological observations of the brain indicate evidence of inflammation and greater tissue pathology in Cd-or Mn-exposed mice compared to control animals . Meningitis and vascular congestion was seen in virus infected mice in all the metal treated groups, and further, the perivascular inflammation appeared earlier in treated mice compared to control . Encephalitis was maximum in Cd pretreated mice . Widespread environmental contamination of metals and the potential for their exposure and subsequent infection of humans or animals is indicative that further studies of these and all other metals are important to understand the effect of environmental pollution on human health. J Matern Fetal Neonatal Med, 2002 Oct, 12(4), 237 - 46 Neutrophil elastase and secretory leukocyte protease inhibitor in prelabor rupture of membranes, parturition and intra-amniotic infection; Helmig BR et al.; OBJECTIVE: Neutrophil elastase (NE), a multifunctional serine protease stored in azurophilic granules of mature neutrophils, is capable of intracellular degradation of proteins during phagocytosis and extracellular degradation of connective tissue during an inflammatory process . Secretory leukocyte protease inhibitor (SLPI) is a natural NE inhibitor present in amniotic fluid, fetal membranes and cervical mucus . An imbalance between NE and SLPI has been implicated as a mechanism of abnormal tissue destruction in chronic inflammatory diseases . The purpose of this study was to determine if parturition, premature rupture of the membranes (PROM) and microbial invasion of the amniotic cavity are associated with changes in amniotic fluid concentrations of NE and SLPI . STUDY DESIGN: Amniotic fluid was retrieved by amniocentesis from 380 patients in the following groups: (1) preterm labor and intact membranes without microbial invasion of the amniotic cavity who delivered at term (n = 13) or prematurely (n = 26), and preterm labor with microbial invasion of the amniotic cavity (n = 9); (2) preterm PROM with (n = 34) and without (n = 51) microbial invasion of the amniotic cavity; and (3) term gestation without microbial invasion of the amniotic cavity with intact membranes not in labor (n = 63), in labor (n = 158), and with rupture of membranes not in labor (n = 26) . Microbial invasion of the amniotic cavity was determined by a positive amniotic fluid culture for micro-organisms including aerobic, anaerobic and Mycoplasma species . NE and SLPI amniotic fluid levels were determined by highly specific and sensitive immunoassays . RESULTS: Preterm PROM was associated with a significant increase in the amniotic fluid concentration of NE . Microbial invasion of the amniotic cavity was associated with a significant increase in the amniotic fluid concentration of NE in women with preterm labor and intact membranes, as well as in women with preterm PROM . Term and preterm parturition was associated with a significant increase in the amniotic fluid concentration of NE . In the absence of microbial invasion of the amniotic cavity, preterm and term PROM were associated with a significant reduction in the amniotic fluid concentration of SLPI . CONCLUSION: Preterm PROM, microbial invasion of the amniotic cavity, and parturition at term and preterm are associated with a significant increase in the amniotic fluid concentration of NE . PROM is associated with a reduced amniotic fluid concentration of SLPI. J Matern Fetal Neonatal Med, 2002 Oct, 12(4), 231 - 6 Protein kinase C stimulates release of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 by human decidual cells; Edwin SS et al.; OBJECTIVE: Increased concentrations of amniotic fluid matrix metalloproteinase (MMP)-9 and tissue inhibitor of metalloproteinase (TIMP)-1 have been observed in the context of premature rupture of membranes (PROM) and microbial invasion of the amniotic cavity . However, the source of the stimuli that contribute to the accumulation of these proteins in amniotic fluid remains to be identified . The present study was conducted to investigate MMP-2, MMP-9 and TIMP-1 secretion by decidual cells in response to activated protein kinase C (PKC) . METHODS: Decidual cells were isolated from term placentae, grown to confluence and incubated with control media or 10(-11) to 10(-8) mol/l concentrations of phorbol 12-myristate 13-acetate (PMA) . Concentrations of MMP-2, MMP-9 and TIMP-1 in the culture supernatant were determined using sensitive and specific immunoassays . Substrate zymography was conducted to confirm MMP-9 assays . RESULTS: PMA induced a concentration-dependent stimulation of release of MMP-9 (control vs . PMA l0(-9) and 10(-8) mol/l; p < 0.01) and TIMP-1 (control vs . PMA 10(-9) and 10(-8) mol/l; p < 0.001), but not MMP-2 . A direct positive correlation was observed between MMP-9 and TIMP-1 release (r = 0.645; p < 0.001) . Substrate zymography confirmed increased release of MMP-9 in response to PMA (control vs . PMA 10(-8) and PMA 10(-7) mol/l; p < 0.01) . CONCLUSIONS: Activation of PKC within the decidua will result in enhanced MMP-9 release, which upon activation could contribute to degradation of matrices within fetal membranes leading to PROM. Appl Environ Microbiol, 2003 Feb, 69(2), 1320 - 4 Dibenzyl sulfide metabolism by white rot fungi; Van Hamme JD et al.; Microbial metabolism of organosulfur compounds is of interest in the petroleum industry for in-field viscosity reduction and desulfurization . Here, dibenzyl sulfide (DBS) metabolism in white rot fungi was studied . Trametes trogii UAMH 8156, Trametes hirsuta UAMH 8165, Phanerochaete chrysosporium ATCC 24725, Trametes versicolor IFO 30340 (formerly Coriolus sp.), and Tyromyces palustris IFO 30339 all oxidized DBS to dibenzyl sulfoxide prior to oxidation to dibenzyl sulfone . The cytochrome P-450 inhibitor 1-aminobenzotriazole eliminated dibenzyl sulfoxide oxidation . Laccase activity (0.15 U/ml) was detected in the Trametes cultures, and concentrated culture supernatant and pure laccase catalyzed DBS oxidation to dibenzyl sulfoxide more efficiently in the presence of 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate) (ABTS) than in its absence . These data suggest that the first oxidation step is catalyzed by extracellular enzymes but that subsequent metabolism is cytochrome P-450 mediated. Appl Environ Microbiol, 2003 Feb, 69(2), 1299 - 304 High rate of uptake of organic nitrogen compounds by Prochlorococcus cyanobacteria as a key to their dominance in oligotrophic oceanic waters; Zubkov MV et al.; Direct evidence that marine cyanobacteria take up organic nitrogen compounds in situ at high rates is reported . About 33% of the total bacterioplankton turnover of amino acids, determined with a representative {(35)S}methionine precursor and flow sorting, can be assigned to Prochlorococcus spp . and 3% can be assigned to Synechococcus spp . in the oligotrophic and mesotrophic parts of the Arabian Sea, respectively . This finding may provide a mechanism for Prochlorococcus' competitive dominance over both strictly autotrophic algae and other bacteria in oligotrophic regions sustained by nutrient remineralization via a microbial loop. Appl Environ Microbiol, 2003 Feb, 69(2), 827 - 34 In-field spatial variability in the degradation of the phenyl-urea herbicide isoproturon is the result of interactions between degradative Sphingomonas spp . and soil pH; Bending GD et al.; Substantial spatial variability in the degradation rate of the phenyl-urea herbicide isoproturon (IPU) {3-(4-isopropylphenyl)-1,1-dimethylurea} has been shown to occur within agricultural fields, with implications for the longevity of the compound in the soil, and its movement to ground- and surface water . The microbial mechanisms underlying such spatial variability in degradation rate were investigated at Deep Slade field in Warwickshire, United Kingdom . Most-probable-number analysis showed that rapid degradation of IPU was associated with proliferation of IPU-degrading organisms . Slow degradation of IPU was linked to either a delay in the proliferation of IPU-degrading organisms or apparent cometabolic degradation . Using enrichment techniques, an IPU-degrading bacterial culture (designated strain F35) was isolated from fast-degrading soil, and partial 16S rRNA sequencing placed it within the Sphingomonas group . Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified bacterial community 16S rRNA revealed two bands that increased in intensity in soil during growth-linked metabolism of IPU, and sequencing of the excised bands showed high sequence homology to the Sphingomonas group . However, while F35 was not closely related to either DGGE band, one of the DGGE bands showed 100% partial 16S rRNA sequence homology to an IPU-degrading Sphingomonas sp . (strain SRS2) isolated from Deep Slade field in an earlier study . Experiments with strains SRS2 and F35 in soil and liquid culture showed that the isolates had a narrow pH optimum (7 to 7.5) for metabolism of IPU . The pH requirements of IPU-degrading strains of Sphingomonas spp . could largely account for the spatial variation of IPU degradation rates across the field. Curr Drug Targets Infect Disord, 2003 Mar, 3(1), 66 - 75 New approaches for anti-infective drug discovery: antibiotics, vaccines and beyond; Cheng Q et al.; Infectious disease is the leading cause of death worldwide, and billions of dollars are invested every year in developing anti-infective drugs . In the meantime, resistant bacteria are on the steady rise and render many once effective drugs useless . The tremendous funding and the urgent need to treat the resistant bacterial infections lead to the rapid progress on development of new drugs and potential new drug targets . New discoveries are being made that increase our understanding of microbial pathogenesis . Technological advancement is also being made to accelerate the drug discovery process . This review will mainly focus on discussing novel strategies on the development of antibiotics and vaccines for treating bacterial infections . Details of how some of the emerging technologies such as genomics and bioinformatics are accelerating the drug discovery process will be highlighted . Newly emerging concepts in controlling bacterial infections such as the use of probiotics and enzybiotics will also be briefly described. Curr Drug Targets Immune Endocr Metabol Disord, 2003 Mar, 3(1), 1 - 22 Controlled modulation of inflammatory, stress and apoptotic responses in macrophages; Malyshev IY et al.; An outstanding question of current immunology is to define the mechanisms by which microbial products influence the immunopathologic host response elements in the early stages of infection . Macrophages are now well recognized to have a critical role in both innate and acquired immunity . In order to adjust promptly to continuous changes in microenvironment and maintain the immunologic balance, macrophages adequately respond by activating one of the numerous immunologic programs . However, sustained macrophage activation and excessive production of inflammatory mediators can perpetuate the numerous pathological processes and contribute to induction of stress response and even apoptosis . Therefore, selective modulation of macrophage activity represents an important strategy for prevention and treatment of inappropriate inflammatory responses in order to minimize the unwanted side-effects of the immunity . Macrophages can be selectively reprogrammed for a specific phenotype of immune response, e.g . cytokine or nitric oxide (NO), by relatively short-term exposure of the cells to substimulatory concentrations of different microbial components, including LPS . These LPS-dependent reprogramming effects are mediated by IFN-gamma-independent autocrine cytokine regulatory mechanisms that also controlled at the transcriptional level . Furthermore, LPS reprogrammed macrophages exhibit differential capacity to resist experimentally induced apoptosis and to produce heat shock proteins . Complete analysis of, and appreciation for, the immunoregulatory mechanisms implicated in LPS-dependent reprogramming of immune responses in macrophages can be expected to increase our understanding of the host innate response, as well as allow investigators to utilize emerging immunologic technologies in effective treatment of infections and chronic inflammatory diseases. Curr Med Chem, 2003 Mar, 10(6), 505 - 24 Beta-propellers: associated functions and their role in human diseases; Pons T et al.; The beta-propeller fold appears as a very fascinating architecture based on four-stranded antiparallel and twisted beta-sheets, radially arranged around a central tunnel . Similar to the alpha/beta-barrel (TIM-barrel) fold, the beta-propeller has a wide range of different functions, and is gaining substantial attention . Some proteins containing beta-propeller domains have been implicated in the pathogenesis of a variety of diseases such as cancer, Alzheimer, Huntington, arthritis, familial hypercholesterolemia, retinitis pigmentosa, osteogenesis, hypertension, and microbial and viral infections . This article reviews some aspects of 3D structure, amino acids sequence regularities, and biological functions of the proteins containing beta-propeller domains . Major emphasis has been laid on beta-propellers whose functions are associated to human diseases . Recent research efforts reported in the fields of protein engineering, drug design, and protein structure-function relationship studies, concerning the beta-propeller architecture, have also been discussed. Phys Rev Lett . 2003 Jan 10;90(1):018101 . Epub 2003 Jan 03. Minimal model for genome evolution and growth; Hsieh LC et al.; Textual analysis of typical microbial genomes reveals that they have the statistical characteristics of a DNA sequence of a much shorter length . This peculiar property supports an evolutionary model in which a genome evolves by random mutation but primarily grows by random segmental duplication . That genomes grew mostly by duplication is consistent with the observation that repeat sequences in all genomes are widespread and intragenomic and intergenomic homologous genes are preponderant across all life forms. Tidsskr Nor Laegeforen, 2002 Dec 10, 122(30), 2891 - 4 {Mobilization of mesenchymal infection defense}; Seljelid R et al.; BACKGROUND: Most infections are arrested in epithelial and superficial connective tissues long before antibodies and antigen specific killer cells have been induced; i.e . before the specific defence system has had time enough to come to the rescue . Microbial substances that activate and modulate this non-specific first-line defence in and near the body surfaces may enhance disease resistance, mainly by stimulating the production of anti-microbial substances by epithelia and by local activation of tissue macrophages . MATERIAL AND METHODS: There are many different microbial substances that can activate macrophages . Beta-1,3-glucans from yeast and mushrooms are the most obvious candidates for pharmaceutical development because their chemical composition and mode of action has been clarified in great detail . RESULTS: Beta-1,3-glucans in purified form provide efficient protection of animals against infections by virus, bacteria, fungi and parasites . Such enhanced protection is obtained after injection as well as after oral or mucosal administration . Beta-1,3-glucans also counteract the toxic effects of bacterial endotoxins and enhance the body's capacity to destroy cancer cells . INTERPRETATION: Activation of non-specific immunity in epithelia and in connective tissues by purified microbial substances corresponds to early events in a natural infection process and renders animals more resistant to infections . This way to enhance resistance to microbial infections has been applied with success in animal husbandry with beta-1,3-glucans administrated orally or onto mucosal surfaces . Corresponding use in human medicine is a realistic possibility, in addition to the use of microbial immune modulators as adjuvants in mucosal vaccines. Zhongguo Yi Xue Ke Xue Yuan Xue Bao, 1999 Apr, 21(2), 140 - 5 {Establishment of a microbial assay for screening anti-human immunodeficiency virus type-1 protease inhibitors}; Leng Q et al.; OBJECTIVE: This study was to establish a microbial assay of human immunodeficiency virus type-1 protease (HIV-1 PR) activity for screening anti-HIV PR inhibitors . METHODS: A 24 bp synthetic oligonucleotide fragment that encodes the HIV-1 PR recognition sequence was inserted into the tetr gene of pBR322 (mtetr) . Escherichia coli containing HIV-1 PR expression vector-pPOLO was transformed with pACYC184M containing modified mtetr gene . The transformant could express both HIV-1 PR and the modified Tet protein . RESULTS: The growth of engineered E . coli was prevented in the presence of tetracycline because the resistance Tet protein was degraded by HIV-1 PR . However inhibition of the HIV-1 PR restored tetracycline resistance . 31 chemical synthetic compounds were tested by the microbial assay . CONCLUSIONS: A microbial assay method of HIV-1 PR activity was established through a engineered E . coli . 5 mumol/L saqunavir-a special HIV-1 PR inhibitor showed inhibitory effect on the engineered E . coli . That means this model could be used as a initial screening model for anti-HIV PR agents. J Microencapsul, 2002 Nov-Dec, 19(6), 797 - 809 Reacetylated chitosan microspheres for controlled delivery of anti-microbial agents to the gastric mucosa; Portero A et al.; The high aqueous solubility of chitosan restricts the utility of chitosan microspheres for gastric drug delivery . This paper describes the preparation of reacetylated chitosan microspheres with suitable properties for the controlled release of active anti-microbial agents, such as amoxycillin and metronidazole, in the gastric cavity . Two different microencapsulation approaches were developed and optimized in order to encapsulate hydrophilic (amoxycillin) and hydrophobic (metronidazole) compounds efficiently . The reacetylated chitosan microspheres exhibited a controlled water swelling capacity and gelified at acidic pH, resulting in prolonged release of the encapsulated antibiotics . The reacetylation time was found to be a key factor that affects not only drug release, but also encapsulation efficiency and anti-microbial activity of the encapsulated compound . The last two parameters were also dependent on drug solubility in the reacetylating agent . Using short reacetylation time periods, it was possible to efficiently control the release of both hydrophilic and lipophilic antibiotics while maintaining their activity against different bacteria . Consequently, reacetylated chitosan microspheres are promising vehicles for the controlled delivery of anti-microbial agents to the gastric cavity and, hence, for the eradication of Helicobacter pylori, a pathogen strongly associated with gastric ulcers and possibly gastric carcinoma. Cytotherapy, 2002, 4(6), 539 - 49 Catastrophic failures of freezing bags for cellular therapy products: description, cause, and consequences; Khuu HM et al.; BACKGROUND: Container integrity is critical for maintaining sterility of cryopreserved cellular therapy products . We investigated a series of catastrophic bag failures, first noticed in early 2001 . METHODS: Process records were reviewed for all PBPC and lymphocyte products cryopreserved in bags from January 2000 through April 2002 . Patient charts were also reviewed . RESULTS: One thousand two hundred and four bags were removed from storage for infusion to 261 patients . All products had been cryopreserved in Cryocyte poly(ethylene co-vinyl acetate) (EVA) bags in either 10% DMSO or 5% DMSO and 6% pentastarch . Product volumes were 25-75 mL, and bags were stored with overwrap bags in a liquid nitrogen tank . From January 2000 to April 2001, failure occurred in 10 of 599 (1.7%) bags . From May 2001 to April 2002, 58 of 605 (9.6%) bags failed, typically with extensive fractures that were visible before thaw . Of the 58 that failed, 24 were salvaged by aseptic methods and infused to patients under antibiotic coverage; 10 of those 24 (42%) had positive bacterial cultures . Bag failures were not related to product type, cryoprotectant solution, liquid versus vapor storage, or freezer location . Failures were linked to use of four Cryocyte bag lots manufactured in 2000 and 2001 . After replacing these lots with a 1999 Cryocyte lot and with KryoSafe polyfluoroethylene polyfluoropropylene (FEP) bags, no more failures occurred in 75 and 102 bags, respectively, thawed through April 2002 . DISCUSSION: High rates of bag failure were associated with four Cryocyte bag lots . No serious adverse patient effects occurred, but bag failures led to microbial contamination, increased product preparation time, increased antibiotic use, and increased resource expenditure to replace products. Br J Nutr, 2003 Jan, 89(1), 97 - 103 Effects of feed intake on composition of sheep rumen contents and their microbial population size; Rodriguez CA et al.; The present study was conducted to determine the effect of feed intake on the composition of the rumen contents of sheep and on their bacterial densities . Whole rumen contents were sampled after a period of continuous inter-rumen infusion of 15NH3 from four rumen-cannulated wethers successively fed on a hay-concentrate diet (2:1, w/w on a DM basis) at two rates of feed intake: 40 and 80 g DM/kg body weight0.75 . Total weight and chemical composition of rumen contents, as well as the distribution by size and chemical composition of particles, were determined . The populations of bacteria associated with the liquid (liquid-associated bacteria, LAB) and solid (solid-associated bacteria, SAB) fractions of rumen digesta and the distribution of SAB according to feed particle size were also examined . The greater feed intake caused an increase in the mass of the rumen contents, while its chemical composition did not change, except for a higher content of organic matter (P=0.023) . The distribution of feed particles by size was similar at both levels of intake . The concentrations of neutral- and acid-detergent fibre in feed particles decreased and those of total, dietary, and microbial N increased, both with a quadratic response (P=0.001), as particle size decreased . The proportion of LAB in the microbial biomass of rumen digesta reached only 8.0 % . This proportion and the density of LAB were unaffected by the level of feed intake, whereas an apparent reduction (10.4 %) occurred with the SAB biomass in whole rumen contents . A systematic, but not significant, reduction (mean value 11.9 %) in the level of microbial colonisation in the different particle fractions with the increase of feed intake was also observed. Hautarzt, 2003 Jan, 54(1), 45 - 52 Epub 2002 Dec 20. {Significance of immunologic tolerance in dermatology}; Steinbrink K et al.; The immunological tolerance processes enable the organism to distinguish between self and non-self and are, therefore, critical for an efficient immune system . Exogenous or endogenous factors that disturb tolerance mechanisms induce uncontrolled activation of the immune system and the development of autoimmune diseases . In the field of dermatology, the most relevant autoimmune diseases are connective tissue diseases and autoimmune bullous skin disorders . In contrast, increased activity of tolerance shuts down parts of the normal immune response and thus facilitates the development of neoplasia and microbial infections in the skin and other organs . Immunological mechanisms for the induction of tolerance have been studied with the help of experimental models of tolerance to contact allergens . T- and B-cells, as well as antigen presenting cells, in particular dendritic cells, are involved in the immunological mechanisms of tolerance . The modification of autologous immune cells of patients with malignant tumors, allergic and autoimmune diseases might have potential for the development of new therapies. Curr Microbiol, 2003 Mar, 46(3), 233 - 4 A thermodynamic interpretation of the Monod equation; Liu Y et al.; The Monod equation for microbial growth is purely empirical, and the theoretical basis of this model stays unclear . Similar to any chemical reactions, overall microbial growth process is dependent upon the changes in free energy . This study showed that the Monod equation could be interpreted in a thermodynamic sense very well . It was probably for the first time demonstrated that the Monod constant ( K(s)) was inversely related to the equilibrium constant of the overall microbial growth process. Genome Res, 2003 Feb, 13(2), 206 - 15 A microarray-based antibiotic screen identifies a regulatory role for supercoiling in the osmotic stress response of Escherichia coli; Cheung KJ et al.; Changes in DNA supercoiling are induced by a wide range of environmental stresses in Escherichia coli, but the physiological significance of these responses remains unclear . We now demonstrate that an increase in negative supercoiling is necessary for transcriptional activation of a large subset of osmotic stress-response genes . Using a microarray-based approach, we have characterized supercoiling-dependent gene transcription by expression profiling under conditions of high salt, in conjunction with the microbial antibiotics novobiocin, pefloxacin, and chloramphenicol . Algorithmic clustering and statistical measures for gauging cellular function show that this subset is enriched for genes critical in osmoprotectant transport/synthesis and rpoS-driven stationary phase adaptation . Transcription factor binding site analysis also supports regulation by the global stress sigma factor rpoS . In addition, these studies implicate 60 uncharacterized genes in the osmotic stress regulon, and offer evidence for a broader role for supercoiling in the control of stress-induced transcription. Nat Rev Mol Cell Biol, 2003 Feb, 4(2), 95 - 104 NALPs: a novel protein family involved in inflammation; Tschopp J et al.; A newly discovered family of cytoplasmic proteins--the NALPs--has been implicated in the activation of caspase-1 by the Toll-like receptors (TLRs) during the cell's response to microbial infection . Like the structurally related apoptotic protease-activating factor-1 (APAF-1), which is responsible for the activation of caspase-9, the NALP1 protein forms a large, signal-induced multiprotein complex, the inflammasome, resulting in the activation of pro-inflammatory caspases. Wei Sheng Yan Jiu, 2001 Nov, 30(6), 357 - 8, 361 {Survey on the microbial pollution of pure water in water dispenser and its disinfection}; Chen X et al.; The counts of bacteria, total coliforms and fungi of pure water in the water dispensers were detected and the regular pattern of bacterial reproduction in water were studied after the dispensers and water were disinfected . The results showed the bacterial pollution in the water dispensers was serious and the bacteria counts increased with the time of using dispensers lasted . The disinfectant could killed effectively the bacteria in the water sources but it could not control the new bacteria polluted and reproduced while the water dispensers were in use . It was suggested that the bacteria polluted from air should be prevented by taking effective methods and selecting appropriate organic materials for making dispensers. Arch Microbiol, 2003 Jan-Feb, 179(2), 75 - 82 Epub 2002 Dec 06. Relevance of airborne fungi and their secondary metabolites for environmental, occupational and indoor hygiene; Fischer G et al.; Airborne fungal contaminants are increasingly gaining importance in view of health hazards caused by the spores themselves or by microbial metabolites . In addition to the risk for infection, the allergenic and toxigenic properties, as well as the inflammatory effects are discussed in this review as possible health impacts of bioaerosols . A major problem is the lack of threshold values for pathogenic and non-pathogenic fungi, both in the workplace and in outdoor air . While the relevance of mycotoxins has been intensely studied in connection with contamination of food and feed, the possible respiratory uptake of mycotoxins from the air has so far not been sufficiently taken into account . Toxic secondary metabolites are expected to be present in airborne spores, and may thus occur in airborne dust and bioaerosols . Potential health risks cannot be estimated reliably unless exposure to mycotoxins is determined qualitatively and quantitatively . Microbial volatile organic compounds (MVOC) have been suggested to affect human health, causing lethargy, headache, and irritation of the eyes and mucous membranes of the nose and throat . The production of MVOC by fungi has been discussed in connection with domestic indoor microbial pollution, but the relevance of fungal metabolites in working environments remains insufficiently studied. J Mol Biol, 2003 Feb 14, 326(2), 517 - 27 Crystal structures of Tritrichomonasfoetus inosine monophosphate dehydrogenase in complex with substrate, cofactor and analogs: a structural basis for the random-in ordered-out kinetic mechanism; Prosise GL et al.; The enzyme inosine monophosphate dehydrogenase (IMPDH) is responsible for the rate-limiting step in guanine nucleotide biosynthesis . Because it is up-regulated in rapidly proliferating cells, human type II IMPDH is actively targeted for immunosuppressive, anticancer, and antiviral chemotherapy . The enzyme employs a random-in ordered-out kinetic mechanism where substrate or cofactor can bind first but product is only released after the cofactor leaves . Due to structural and kinetic differences between mammalian and microbial enzymes, most drugs that are successful in the inhibition of mammalian IMPDH are far less effective against the microbial forms of the enzyme . It is possible that with greater knowledge of the structural mechanism of the microbial enzymes, an effective and selective inhibitor of microbial IMPDH will be developed for use as a drug against multi-drug resistant bacteria and protists . The high-resolution crystal structures of four different complexes of IMPDH from the protozoan parasite Tritrichomonas foetus have been solved: with its substrate IMP, IMP and the inhibitor mycophenolic acid (MPA), the product XMP with MPA, and XMP with the cofactor NAD(+) . In addition, a potassium ion has been located at the dimer interface . A structural model for the kinetic mechanism is proposed. Carbohydr Res, 2003 Feb 14, 338(5), 423 - 31 Isolation and characterisation of the homogalacturonan from type II cell walls of the commelinoid monocot wheat using HF-solvolysis; Wietholter N et al.; In contrast to the typical type I cell wall of the dicot plants, the type II cell wall of the commelinoid monocot plants is known to be relatively poor in pectins . Assuming a critical role for the remaining pectins in terms of cell wall architecture and/or as a reservoir of signalling molecules, we have compared different protocols for the isolation of the main pectin polymer, homogalacturonan, from wheat leaf cell walls . Pectin was detected in these cell walls immunochemically using the monoclonal antibodies JIM5 and JIM7, and biochemically by monosaccharide analysis . The Ca(++)-chelators CDTA and imidazole extracted a pectin rich fraction from isolated cell walls which was however contaminated with significant amounts of hemicelluloses . Pretreatment of the cell walls with anhydrous hydrogen fluoride at controlled low temperatures followed by HF/ether- and water-extraction prior to imidazole-extraction of pectins yielded a purer homogalacturonan fraction . The near absence of rhamnosyl residues proved that the isolated homogalacturonan fraction was free of rhamnogalacturonans . If HF-solvolysis was performed at -23 degrees C, the resulting homogalacturonan had a degree of methyl esterification identical to that of the pectins in the initial wheat cell wall . The antibodies JIM5 and JIM7 as well as PAM1 and LM5 proved that the isolated homogalacturonan had a low methyl ester content, was polymeric and free of galactan side chains . We can thus isolate native homogalacturonan from the type II wheat cell walls with the original in muro pattern of methyl esterification still intact, to further investigate e.g., its degradability by plant or microbial pectic enzymes. J Periodontal Res, 2003 Feb, 38(1), 104 - 8 Relationship between fibrinolytic activity and gingival inflammatory reaction in young individuals; Olofsson A et al.; The fibrinolytic system (the plasminogen activating system) is involved in several physiological and pathological processes . Through the transformation of plasminogen to the aggressive broad spectrum protease plasmin, potent enzymatic activity is released . Plasmin acts directly on connective tissue components, and indirectly by activating proforms of the metalloproteinases . The destructive potential of the fibrinolytic system may thus be of importance for the initiation and progression of periodontal diseases . Earlier studies have shown high concentrations of the plasminogen activator t-PA and its inhibitor PAI-2 in gingival crevicular fluid (GCF) as well as enhanced concentrations in areas of gingival inflammation . The aim of this study was to investigate a possible relationship between the gingival inflammatory reactivity and the fibrinolytic activity in gingival crevicular fluid . Thirty-one young individuals took part in the study . Gingival Index scores and Plaque Index scores were assessed and used to formulate a score expressing an individuals' inflammatory response to microbial plaque levels (Relative G/P score) . The fibrinolytic activity of GCF was assessed with a fibrin gel lysis assay, and the levels of t-PA and PAI-2 were assayed with ELISAs . All samples showed fibrinolytic activity . A positive correlation between the fibrinolytic activity and Relative G/P score was found . Thus, in individuals with an enhanced reactivity to dental plaque, a higher plasminogen activating activity in GCF was seen . This indicates a higher potential for tissue proteolysis in these individuals, possibly facilitating spread and deeper involvement of the lesions. Environ Microbiol, 2003 Feb, 5(2), 92 - 102 Physiological and molecular characterization of anaerobic benzene-degrading mixed cultures; Ulrich AC et al.; Nine distinct anaerobic benzene-degrading cultures were enriched from sediment samples from four different sites . These cultures used nitrate, sulphate or CO2 as electron acceptors . The shortest doubling times were observed in nitrate-reducing cultures, although cell yield was lowest in these cultures . The highest substrate concentration utilized and maximum absolute rates of benzene degraded (in micro M day-1) were observed in methanogenic cultures . The microbial compositions of a methanogenic and nitrate-reducing culture were determined from a clone library of 16S rRNA genes . Five Bacterial 16S rRNA sequences, one of which resembled a clone previously found in a sulphate-reducing, benzene-degrading culture and four Archaeal 16S rRNA sequences were identified in a methanogenic culture . Four Bacterial and no Archaeal 16S rRNA sequences were identified in a nitrate-reducing culture . The relative abundance of the four nitrate-reducing putative species was determined by slot blot hybridization . Two green sulphur bacteria together formed 52% of the clone library, but were found to be less than 4% of the culture by slot blot analysis . One of the cloned 16S rRNA gene sequences comprised 70% of the culture and was phylogenetically 93% similar to both Azoarcus and Dechloromonas species, which have been shown to degrade aromatic compounds, including benzene, under nitrate-reducing conditions. Pest Manag Sci, 2003 Jan, 59(1), 114 - 8 Dissipation of the herbicide dithiopyr in soil and residues in wheat (Triticum aestivum L) grain under Indian tropical conditions; Saikia N et al.; Dissipation of dithiopyr in soil was monitored after application to wheat crop as pre- or post-emergence applications at two rates, viz 100 and 200 g AI ha(-1) . The level of dithiopyr in the soil was assessed by gas chromatography, and its disappearence was found to follow a first-order decay curve irrespective of rate or method of application . The half-life in soil ranged between 17.3 and 25.0 days and residues at harvest (150 days after application) ranged between 4.0 and 8.8% of amounts applied . Investigation of microbial degradation of dithiopyr was conducted in minimal salt and Czapek Dox media in which 80% of the compound degraded within 15 days . Residues were not detected in wheat grain at harvest. J Egypt Soc Parasitol, 2001 Apr, 31(1), 21 - 35 Laboratory evaluation of a Helicosporidium sp . (Protozoa: Helicosporida) as an agent for the microbial control of mosquitoes; Seif AI et al.; A Helicosporidium sp . isolated from diseased larvae of Culex pipiens L . in Egypt was assessed for its microbial control potential against mosquito larvae . Dose infectivity and dose mortality of the pathogen was determined for each larval instar of Cx . pipiens . Helicosporidium sp was infectious to Aedes caspius (Pallas), C . antennatus (Becker), C . perexiguus Theobald, and Culiseta longiareolata (Macquart) . Using 24-hr exposures of first instar larvae to a range of concentrations between 50 and 5 x 10(3) spores/ml, Cx . antennatus and Cx . perexiguus were more susceptible than Cx . pipiens, Ae . caspius and Cs . longiareolata with LC50's of 5 x 10(2), 5 x 10(2), 1.6 x 10(3), 1.4 x 10(3), and 1.7 x 10(3), respectively . Current experiments shown that Helicosporidium spores were tolerant of freezing and storage at 5 degrees C, but not to desiccation . Production trails indicated that up to 6.2 x 10(8) spores could be routinely produced in individual cotton leaf worms, Spodoptera littolaris (Boisd) (Lepidoptera: Noctuidae). Wei Sheng Wu Xue Bao, 2002 Aug, 42(4), 484 - 9 {Asymmetric microbial reduction of organosilyl ketone with immobilized Saccharomyces cerevisiae cells}; Lou W et al.; Asymmetric microbial reduction of acetyltrimethylsilane with immobilized Saccharomyces cerevisiae cells was studied . The effects of shake speeds, substrate concentration, immobilized cell concentration, pH and reaction temperature on the initial reaction rate, maximum yield and optical purity of the product were systematically studied . All above mentioned parameters have important effects on the raction . The optimum shake speed, substrate concentration, immobilized cell concentration, pH and reaction temperature were found to be 150 r/min, 14 mmol/L, 0.15 g/mL, 7.3 and 25 degrees C-30 degrees C respectively for the reaction, under which the maximum yield and optical purity of the product were as high as 84.9% and 90.2% ee. Biotechnol Bioeng, 2003 Mar 30, 81(7), 775 - 82 Bioprocess considerations for expanded-bed chromatography of crude canola extract: sample preparation and adsorbent reuse; Bai Y et al.; Compared to the conventional microbial and mammalian systems, transgenic plants produce proteins in a different matrix . This provides opportunities and challenges for downstream processing . In the context of the plant host Brassica napus (canola), this work addresses the bioprocessing challenges of solid fractionation, resin fouling by native plant components (e.g., oil, phenolics, etc.), hydrodynamic stability, and resin reuse for expanded bed adsorption for product capture . Plant tissue processing and subsequent protein extraction typically result in an extract with a high content of solids containing a wide particle-size distribution . Without removal of larger particles, the column inlet distributor plugged . The larger particles (> 50 microm) were easily removed through centrifugal settling comparable to that attainable with a scroll decanter . The remaining solids did not affect the column performance . Less than 4% of the lipids and phenolics in the fed extract bound to STREAMLINE trade mark DEAE resin, and this small proportion could be satisfactorily removed using recommended clean-in-place (CIP) procedures . Hydrodynamic expansion and adsorption kinetics of the STREAMLINE trade mark DEAE resin were maintained throughout 10 cycles of reuse, as was the structural integrity of the resin beads . No significant accumulation of N-rich (e.g., proteins) and C/O-rich components (e.g., oil and phenolics) occurred over the same period . Gastroenterology, 2003 Feb, 124(2), 521 - 36 The genetics of inflammatory bowel disease; Bonen DK et al.; The inflammatory bowel diseases (IBD) comprise complex genetic disorders, with multiple contributing genes . Linkage studies have implicated several genomic regions as likely containing IBD susceptibility genes, with some observed uniquely in Crohn's disease (CD) or ulcerative colitis (UC), and others common to both disorders . The best replicated linkage region, IBD1, on chromosome 16q contains the CD susceptibility gene, NOD2/CARD15 . NOD2/CARD15 is expressed in peripheral blood monocytes and is structurally related to the plant R proteins, which mediate host resistance to microbial pathogens . Three major coding region polymorphisms within NOD2/CARD15 have been highly associated with CD among patients of European descent . Having one copy of the risk alleles confers a 2-4-fold risk for developing CD, whereas double-dose carriage increases the risk 20-40-fold . All 3 major CD variants exhibit a deficit in NF-kappaB activation in response to bacterial components . Carriage of NOD2/CARD15 risk alleles is associated with ileal location, earlier disease onset, and stricturing phenotype . Other IBD genomic regions include IBD2 on chromosome 12q (observed more in UC), and IBD3, containing the major histocompatibility complex region . A short genomic region has been associated with CD on chromosome 5q, but the precise contributing gene is as yet unidentified . The characterization of additional IBD susceptibility genes could potentially lead to the identification of novel therapeutic agents for IBD, make possible a molecular reclassification of disease, and increase understanding of the contribution of environmental factors (notably, tobacco and the intestinal microbial milieu) to intestinal inflammation. Arch Tierernahr, 2002 Oct, 56(5), 327 - 37 Effect of dietary supplementation with alpha-galactosidase preparation and stachyose on growth performance, nutrient digestibility and intestinal bacterial populations of piglets; Pan B et al.; Two experiments were conducted to investigate the effect of dietary supplementation with alpha-galactosidase preparation and stachyose on growth performance, nutrient digestibility and intestinal bacterial populations of piglets . In Experiment I 72 crossbred piglets were allotted to three treatments, i.e . 1) control (basal) diet, 2) alpha-Gal1 and 3) alpha-Gal5, the basal diet supplemented with 0.01% and 0.05% alpha-galactosidase preparation (alpha-Gal), respectively . Average daily gain, average daily feed intake and feed conversion ratio were not influenced by alpha-Gal supplementation . In Experiment II 72 crossbred weanling piglets were randomly assigned to four treatments with a complete factorial design, i.e . 1) basal diet, 2) basal diet with 0.01% alpha-Gal supplementation, 3) basal diet with 1% stachyose supplementation and 4) basal diet with 1% stachyose and 0.01% alpha-Gal supplementation . Average daily gain and feed conversion ratio were influenced by alpha-Gal and stachyose supplementation . The ileal digestibility of stachyose, raffinose, gross energy and crude protein was improved significantly by alpha-Gal supplementation . The microbial populations in the intestine were modified by both alpha-Gal and stachyose supplementation . The data suggested that stachyose supplementation had an adverse effect on the growth performance of piglets and alpha-Gal supplementation could ameliorate it, especially in younger pigs. Ann Chim, 2002 Nov-Dec, 92(11-12), 1057 - 65 Presence and racemisation of amino acids in calcium oxalate patinas . A case study from the Baptistery in Parma, Italy; Casoli A et al.; The organic material present in oxalate patinas, collected from the stone external surfaces of the Baptistery in Parma (Italy), was analysed by GC-MS . The high racemisation levels determined for several specific amino acids indicate that microbial activity could contribute to the formation and modification of the organic material leading to the calcium oxalate patinas. Eur J Immunol, 2002 Nov, 32(11), 3235 - 45 Combined dendritic cell- and CpG oligonucleotide-based immune therapy cures large murine tumors that resist chemotherapy; Heckelsmiller K et al.; The use of dendritic cells (DC) loaded with tumor antigen is one of the most advanced approaches in cancer immunotherapy . CpG motifs within microbial DNA detected by toll-like receptor 9 are responsible for the favorable properties of CpG oligodeoxynucleotides (ODN) as immune modulators . In this study, mature antigen-pulsed DC or peritumoral injections of CpG ODN, both effective for the treatment of small established tumors, were almost ineffective against large established tumors (1-cm diameter) in a syngeneic murine colon carcinoma model . For large tumors, the antitumor activity of mature antigen-pulsed DC was strongly increased by coinjection of CpG ODN, resulting in a transient control of tumor growth . Rejection of large tumors and long-term cure of mice was achieved by combining injection of antigen-pulsed DC plus CpG ODN at a site distant to the tumor with peritumoral injections of CpG ODN . Depletion of CD8 T cells abrogated the therapeutic activity . Large numbers of DEC-205-positive DC infiltrated the tumor in treated mice . Therapy with 5-fluorouracil and leucovorin was unable to control tumors of the same size . In conclusion, we demonstrate that the immune system, provided that appropriate stimulation with DC and CpG ODN is given, has the potential to cure animals of large solid tumors in situations where even chemotherapy is not efficient. J Toxicol Environ Health A, 2003 Feb 28, 66(4), 345 - 63 Ecotoxicological risks associated with land treatment of petrochemical wastes . III . Immune function and hematology of cotton rats; Wilson J et al.; Landfarming is a widely used method of treating petrochemical waste through microbial bio-degradation . The effects of residual petrochemical contamination on wildlife, especially terrestrial mammals, are poorly understood . The effects of contaminants on the immune system and hematology of cotton rats (Sigmodon hispidus) living on five abandoned petrochemical landfarms (units 1-5) in Oklahoma were studied . Cotton rats were sampled seasonally (summer and winter) from each landfarm and from five ecologically matched reference sites for 2 yr (1998-2000) and returned to the laboratory for immunological and hematological assays . Overall analysis indicated that rats inhabiting landfarms exhibited decreased relative spleen size compared to rats collected from reference sites, with the landfarm at unit 1 showing the greatest reduction . Cotton rats collected from landfarms also had increased hemoglobin, hematocrit, and platelet levels and decreased blood leukocytes during summer . During winter, an increase in the number of popliteal node white blood cells was observed from rats collected on landfarms . No marked difference was detected for lymphocyte proliferation in response to concanavalin A, pokeweed, or interleukin-2 . Lymphokine-activated killer cell lytic ability showed a seasonal pattern, but no treatment differences . No differences between landfarm and reference sites were detected in the hypersensitivity reaction of rats given an intradermal injection of phytohemagluttinin (PHA-P) . Comparisons within individual sites indicated that two sites (units 1 and 3) had the greatest effects on immune function and hematology of cotton rats . The results of this study suggest that residual petrochemical waste affects the immune system and hematology of cotton rats living on abandoned landfarms during summer and is complicated by variation in the contaminants found on individual petroleum sites. Water Res, 2003 Mar, 37(5), 1064 - 70 Impact of UV disinfection on microbially available phosphorus, organic carbon, and microbial growth in drinking water; Lehtola MJ et al.; UV irradiation at a wavelength of 253.7 nm (UV(254)) is commonly used for drinking water disinfection . UV radiation is known to convert organically combined phosphorus to orthophosphate and to degrade natural organic matter . We studied if UV disinfection increases the amount of microbially available forms of organic carbon and phosphorus in drinking waters with different characteristics, and if these changes in water chemical quality could enhance the microbial growth in drinking water . The UV(254) dose (15-50 mWs/cm(2)) used in waterworks reduced the concentration of assimilable organic carbon and the sum of the molecular size fractions . The release of microbially available phosphorus needed higher doses (204 mWs/cm(2)) of UV(254) radiation . Of bacteria in drinking water, 90% were inactivated with UV(254)-irradiation doses below 50 mWs/cm(2) . A high dose (501 mWs/cm(2)) of UV(254) radiation inhibited the microbial growth in water. Dis Aquat Organ, 2002 Dec 10, 52(3), 217 - 31 Continuous in vitro culture of the carpet shell clam Tapes decussatus protozoan parasite Perkinsus atlanticus; Casas SM et al.; Continuous in vitro cultures of the clam Tapes decussatus parasite Perkinsus atlanticus were established from infected gill fragments, infected haemolymph and parasite hypnospores isolated from infected gill fragments following incubation in Ray's fluid thioglycollate medium (RFTM) . No continuous cultures could be initiated from P . atlanticus zoospores . Cultures initiated from hypnospores yielded the highest percentage of continuous cultures (100%, 6/6), followed by cultures initiated from gill fragments (93%, 43/46) and from haemolymph (30%, 3/10) . Failures to establish continuous cultures were due to microbial contamination . The source of parasite influenced the success rate, the time taken to establish cultures and the size of cultured cells . In vitro proliferation of parasite cells was mainly by vegetative multiplication . Zoosporulation, yielding motile biflagellated zoospores, was observed at a low frequency (< 1% of dividing cells) in every culture . Morphology of cultured cells examined with light and transmission electron microscopy corresponded to that of P . atlanticus found in clam tissues . Cultured cells enlarged in RFTM and stained blue-black with Lugol's solution, which are characteristics of the Perkinsus species cells . DNA sequences of the internal transcribed spacer (ITS) region of the ribosomal RNA gene complex matched those of P . atlanticus . All cultures were established in a medium designated JL-ODRP-2A that was similar in composition to the culture medium JL-ODRP-1 originally used to propagate Perkinsus marinus in vitro . Proliferation of P . atlanticus in vitro could be supported by the commercial culture medium (1:2 v/v) DME:Ham's F-12 with fetuin. Wei Sheng Wu Xue Bao, 2001 Aug, 41(4), 505 - 9 {Effect of drying and temperature on the sporulation of Pandora delphacis mycelia gelatinized}; Liu Z et al.; The entomophthoraceous fungus, Pandora delphacis, is a microbial agent highly potential for control of sucking-type insects . In this study, effort was made of gelatinizing the mycelia of the isolate F95129 from submerged culture using polyacrylamide-starch gel pwder and sodium alginate . The resulting film-like gel of the mycelia sporulated very well, indicating that the materials used for gelatinization of the mycelia was biologically compatible with P . delphacis . Then, the gelatinized mycelia were slowly dried in silica chamber at 4, 14, and 24 degrees C, respectively . After 35-day drying and storage in the chamber, however, a 22.7%-92.8% decrease of sporulation capacity occurred in the film-like gels, which then had different water content . The nutrient-supplemented polyacrylamide-starch gel stored at 14 degrees C with water content of 6.1%-7.4% was best for largely decreasing the loss of sporulation capacity among the combinations of gel, nutrition, water content and temperature . The results indicate that it could be possible to choose a desirable gel for longer biological activity of gelatinized P . delphacis mycelia. Se Pu, 1999 Sep, 17(5), 483 - 5 {Analysis of organic acids in human dental plaque by means of gas chromatography/mass spectrometry}; Li J et al.; The purpose of this study was to investigate the composition of plaque fatty acids in the healthy population . The study was made on 10 volunteers over the age of 18 who were divided into three sub-groups (3-4 individuals) . Neither subject exhibited clinical evidence of salivary gland disorder and any medication affecting salivary functions were not used . A sensitive GC/MS method with VG7070E mass spectrometer was developed in our study . The sample separation was carried out on a fused silica capillary column with OV-1 . The column size was 23 m x 0.23 mm . The temperature program was as follows: from 40 degrees C to 120 degrees C fast, then from 120 degrees C to 240 degrees C at 6 degrees C/min . The results showed that there were 14 organic acids and isomers present in plaque . They were C12:0, C14:0, C15:0, C16:0, C16:1, C18:0, C18:1, C18:2, C20:0 and phenylacetic acid, phenylpropionic acid . The higher content of fatty acids in the sample were C16:0, C18:0, and C18:1 . The aromatic acids were detected only in some samples . The odd-numbers fatty acids and aromatic acids were for the first time detected . The origin of organic acids in plaque is an ongoing area of research . Our data clearly identify the bacterial contribution to the organic acids of plaque matrix, which may have a special relationship with bacteria metabolism . The research will help us to understand fatty acids metabolism of dental plaque and to determine their role in the microbial homeostasis of dental plaque. Wei Sheng Wu Xue Bao, 2001 Dec, 41(6), 745 - 9 {Microbial reduction ability of various iron oxides in pure culture experiment}; Qu D et al.; The microbial reduction of ferrihydrite, lepidocrocite, hematite, goethite and aluminum-substituted iron oxides were examined by iron-reducer GS-15 under anaerobic pure culture condition . The results indicated that the ferrihydrite and lepidocrocite can be rapidly reduced by iron-reducer, and the percentage of microbial reduction are respectively 95.4% and 95.8% after 4 days incubation at 25 degrees C . The other iron oxides like hematite, Al-hematite, goethite and Al-goethite are very difficult to reduce during short-term incubation . The amounts of Fe(III)-reducing, Fe(II)-producing and Fe-reducing calculated b