Bacteria

Environ Microbiol, 2002 Jan, 4(1), 42 - 50
Prevalence and microdiversity of Alteromonas macleodii-like microorganisms in different oceanic regions; Garcia-Martinez J et al.; The presence, prevalence and variability of microorganisms related to the species Alteromonas macleodii, a well known culturable gamma-Proteobacterium, has been studied in different seawater samples from diverse geographical locations, in both the Northern and Southern hemispheres, and tested with two molecular techniques (rRNA hybridization and gene cloning and sequencing) . Results show that A . macleodii-like microorganisms are present in high proportions in North Atlantic and, especially, Mediterranean waters, being higher at deep samples and particle-associated fractions, in agreement with previous findings . In contrast, Southern samples (all from very cold areas near Antarctica) presented no significant hybridization signals . The analysis of the ribosomal ITS (16S-23S internal transcribed spacers) revealed that A . macleodii-like microorganisms from Mediterranean, North Atlantic, Caribbean and Red Sea waters differed in both size and sequence, mostly depending on their geographical origin, with Mediterranean and North Atlantic clones clustering into two main groups whereas Caribbean and Red Sea clones appeared separated.

Transplantation, 2002 Apr 15, 73(7), 1049 - 54
Tacrolimus (FK506) and cyclosporine reduce the uptake and transport of particles into rabbit Peyer's patches; Fujimura Y et al.; Tacrolimus (FK) and cyclosporine (Cs) are potent immunosuppressants that effectively prevent the rejection of transplanted organs including liver and small intestine . Our study examined the effects of these immunosuppressants on Peyer's patches, which play an important role in mucosal immune response through uptake and transport of enteric microorganisms and macromolecules in the gut-associated lymphoid tissues . After administration of FK and Cs, we assessed changes in lymphoid follicle structure and quantified the uptake and transport of particles in the follicle associated epithelium (FAE) including M cells, using fluorescent latex microspheres in rabbit Peyer's patches . Rabbits, five in each group, received oral administration of FK (3.2 mg/kg), Cs (10 mg/kg), or phosphate-buffered saline daily for 7 days . After 2 days of withdrawal, rabbits were anesthetized, and received injections with 2 ml of the suspension of 0.5-microm fluorescent microspheres (1010/ml) into ligated intestinal segments containing Peyer's patches . After 2 hr of gentle agitation, segments were removed, rinsed, fixed with periodate-lysine-2% paraformaldehyde, frozen, and sections were stained with fluorescent phalloidin to label brush border actin filaments . The size of the lymphoid follicles in each group was measured under a light microscope . The number of microspheres in follicles was assessed in graphically defined areas of follicles from each group . In addition, immunohistochemical analysis of CD43 and MHC-II positive cells in FAE of lymphoid follicles of each group was performed . FK and Cs significantly reduced the height of lymphoid domes and the height and width of follicles, as compared to those of controls . In both FK and Cs groups, the numbers of microspheres that adhered, were taken up and were transported into lymphoid follicles were smaller than in controls, indicating that their movement rates into deep layers were markedly reduced . Furthermore, FK and Cs reduced the mean numbers of CD43 and MHC-II positive cells in FAE per unit area (mm2) as compared with controls . These findings suggest that FK and Cs may produce immunosuppressive effects, at least in part, through reduction of the uptake and transport of particles into Peyer's patches, and by reduction of the number of immunoreactive cells in FAE of Peyer's patches.

Curr Opin Infect Dis, 2001 Aug, 14(4), 461 - 6
Infection control in developing countries; Starling C; Infection control activities in developing countries have increased substantially during the last decade, particularly in certain regions of the world . Pressure to improve hospital care cost-effectiveness, the emergence of highly resistant microorganisms, the perception of healthcare worker occupational hazards, and public claims for improved health assistance quality have been important factors responsible for this development . A major challenge for infection control personnel in developing countries is to sustain this development.

Curr Opin Infect Dis, 2001 Aug, 14(4), 437 - 42
Nosocomial transmission of antibiotic-resistant microorganisms; Harbarth S; The spread of nosocomially acquired, antibiotic-resistant microorganisms has grown dramatically over the past 20 years and has increasingly attracted the attention of academic inquiry . This article reviews recent epidemiological data about the trends and patterns of nosocomial transmission, presents clinical studies investigating mechanisms of cross-transmission, and discusses recent literature on preventive strategies aimed at reducing hand-carriage of antibiotic-resistant pathogens.

Curr Opin Infect Dis, 2000 Apr, 13(2), 103 - 107
Retroviruses and psoriasis; Mallon E; Psoriasis occurs with at least undiminished frequency in HIV infected individuals . The behaviour of psoriasis in HIV disease is of interest, both in terms of pathogenesis and therapy, because of the background of profound immunodysregulation . It is paradoxical that, while drugs that target T lymphocytes are effective in psoriasis, the condition should be exacerbated by HIV infection . Antiretroviral therapy may improve psoriasis in tandem with improvement in the overall clinical and virological condition of the patient . The aetiopathogenesis of psoriasis is unknown but genetic and environmental factors are thought to be involved . There are controversial issues regarding the immunological basis of psoriasis and the role of CD4+ versus CD8+ T lymphocytes . Current opinion favours an autoimmune basis for psoriasis, although the precipitating activating signal(s) within psoriatic plaques remains unknown . The immunodysregulation resulting from HIV infection may trigger psoriasis in those genetically predisposed by the Cw*0602 allele . Since CD8+ T cells recognize antigen in the context of class I molecules, the identification of a human leucocyte antigen class I association in HIV-associated psoriasis strengthens the argument for an important role for CD8+ T lymphocytes in the immunopathogenesis of psoriasis . HLA-Cw*0602 could act as a cross-reactive target for cytotoxic T lymphocytes responding to processed peptides from microorganisms . Human retrovirus-5 is a recently described, partially characterized retrovirus and has been implicated in the pathogenesis of psoriatic arthropathy but not psoriasis.

Food Addit Contam, 2002, 19 Suppl, 178 - 84
Determination of microbial contamination of plastic cups for dairy products and utilization of electron beam treatment for sterilization; Tacker M et al.; Packaging materials are often considered a critical control point in HACCP systems of food companies . Methods for the determination of the microbial contamination rate of plastic cups, especially for dairy products, must reliably detect single moulds, yeasts or coliforms . In this study, a comparison of a specially adapted coating method, impedance method, direct inoculation and membrane filter technique was carried out to determine contamination with yeasts, moulds, coliforms and total bacterial counts using the appropriate agar in each case . The coating method is recommended for determining yeasts, moulds and coliforms as it allows the localization of the microorganisms as well as the determination of single microorganisms . For total bacterial count, a direct inoculation technique is proposed . The employing of simple measures in the production and during transport of packaging materials, such as dust-prevention or tight sealing in polyethylene bags, heavily reduces microbial contamination rates of packaging material . To reduce contamination rates further, electron beam irradiation was applied: plastic cups sealed in polyethylene bags were treated with 4-5 kGy, a dose that already leads to sterile polystyrene and polypropylene cups without influencing mechanical characteristics of the packaging material.

An Acad Bras Cienc, 2002 Mar, 74(1), 151 - 70
Secondary metabolites from marine microorganisms; Kelecom A; After 40 years of intensive research, chemistry of marine natural products has become a mature field . Since 1995, there are signals of decreased interest in the search of new metabolites from traditional sources such as macroalgae and octocorals, and the number of annual reports on marine sponges stabilized . On the contrary, metabolites from microorganisms is a rapidly growing field, due, at least in part, to the suspicion that a number of metabolites obtained from algae and invertebrates may be produced by associated microorganisms . Studies are concerned with bacteria and fungi, isolated from seawater, sediments, algae, fish and mainly from marine invertebrates such as sponges, mollusks, tunicates, coelenterates and crustaceans . Although it is still to early to define tendencies, it may be stated that the metabolites from microorganisms are in most cases quite different from those produced by the invertebrate hosts . Nitrogenated metabolites predominate over acetate derivatives, and terpenes are uncommon . Among the latter, sesquiterpenes, diterpenes and carotenes have been isolated; among nitrogenated metabolites, amides, cyclic peptides and indole alkaloids predominate.

FEBS Lett, 2002 Apr 10, 516(1-3), 67 - 70
Nicotine-modulated formation of spiroiminodihydantoin nucleoside via 8-oxo-7,8-dihydro-2'-deoxyguanosine in 2'-deoxyguanosine-hypochlorous acid reaction; Suzuki T et al.; Hypochlorous acid (HOCl) is generated by myeloperoxidase of activated neutrophils which kill invading microorganisms, but also cause DNA damage in inflamed tissues . We report here that spiroiminodihydantoin nucleoside (dS), a further oxidized product of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG), is formed, in addition to 8-chloro-2'-deoxyguanosine and 8-oxo-dG, by reaction of 2'-deoxyguanosine with HOCl . Presence of low concentrations of nicotine significantly enhanced the yields of these HOCl-modified nucleosides . Our results imply that nicotine may enhance genotoxicity and tissue damage caused by neutrophil activation . dS may also serve as a new biomarker for oxidative DNA damage induced by oxidants such as HOCl.

Comp Biochem Physiol B Biochem Mol Biol, 2002 Mar, 131(3), 371 - 86
Mechanism of biosynthesis of trimethylamine oxide from choline in the teleost tilapia, Oreochromis niloticus, under freshwater conditions; Niizeki N et al.; The mechanism of biosynthesis of trimethylamine oxide (TMAO) from dietary precursors in the teleost tilapia (Oreochromis niloticus) was investigated . Diets supplemented with quaternary ammonium choline, glycine betaine, carnitine or phosphatidylcholine were administered and significant increases in TMAO levels in the muscle were only observed with choline . {Methyl-14C} and {1,2-14C} cholines were given through dietary and intraperitoneal injection routes, but 14C-TMAO was detected only in fish with dietary administration of {methyl-14C} choline . Dietary treatment with {15N} choline resulted in the formation of {15N} TMAO in the muscle . The incorporation of radioactivity into TMAO was also observed both following dietary administration and intraperitoneal injection of {14C} trimethylamine (TMA) . When choline was introduced into the isolated intestine, marked increases in TMA levels occurred . These increases were significantly suppressed in the presence of penicillin . {14C}-TMA derived from {methyl-14C} choline was detected in the cavity of the isolated intestine . The introduction of {15N} choline into the intestinal cavity resulted in the formation of {15N} TMA . TMA mono-oxygenase activities were detected in the liver and kidney . We conclude that tilapia possess the ability to produce TMAO from choline, which is related to intestinal microorganisms and tissue mono-oxygenase under freshwater conditions.

FEMS Microbiol Lett, 2002 Feb 5, 207(2), 111 - 20
Current status of immune mechanisms of killing of intracellular microorganisms; Ismail N et al.; The interaction between intracellular pathogens and the mammalian host follows different pathways that reflect evolved survival mechanisms of both the pathogen and the host to assure each one's own survival . From the host's perspective, different immune mechanisms predominate at different stages of infection . Both phagocytic and non-phagocytic target cells participate in microbial uptake and, in some cases, intracellular destruction . In addition, the development of specific immunity ensures sustained activation of intracellular microbicidal mechanisms in the target cells, and induction of apoptotic or lytic target cell death by cytotoxic T lymphocytes . From the pathogen's perspective, different evasion strategies are employed to counteract host defenses . Understanding microbial survival strategies and the immune mechanisms that result in killing of intracellular pathogens will deepen our insight into the pathogenesis of infection that could be applied towards the development of effective vaccination and immunotherapy.

Prog Lipid Res, 2002 Jul, 41(4), 315 - 67
Natural halogenated fatty acids: their analogues and derivatives; Dembitsky VM et al.; A comprehensive survey has been made of all fatty acids containing halogen atoms covalently bonded to carbon and which are deemed as naturally occurring . Generally thought to be minor components produced by many different organisms, these interesting compounds now number more than 300 . Recent research, especially in the marine area, indicates this number will increase in the future . Sources of halogenated fatty acids include microorganisms, algae, marine invertebrates, and higher plants and some animals . Their possible biological significance has also been discussed

J Agric Food Chem, 2002 Apr 24, 50(9), 2615 - 21
Volatile metabolites from actinomycetes; Scholler CE et al.; Twenty-six Streptomyces spp . were screened for their volatile production capacity on yeast starch agar . The volatile organic compounds (VOCs) were concentrated on a porous polymer throughout an 8-day growth period . VOCs were analyzed by gas chromatography with flame ionization detection and identified or characterized by gas chromatography-mass spectrometry . A total of 120 VOCs were characterized by retention index and mass spectra . Fifty-three compounds were characterized as terpenoid compounds, among which 18 could be identified . Among the VOCs were alkanes, alkenes, alcohols, esters, ketones, sulfur compounds, and isoprenoid compounds . Among the most frequently produced compounds were isoprene, acetone, 1-butanol, 2-methyl-1-propanol, 3-methyl-3-buten-1-ol, 3-methyl-1-butanol, 2-methyl-1-butanol, cyclopentanone, dimethyl disulfide, dimethyl trisulfide, 2-phenylethanol, and geosmin . The relationship between the excretion of geosmin and the production of spores was examined for one isolate . A good correlation between headspace geosmin and the number of spores was observed, suggesting that VOCs could be used to indicate the activity of these microorganisms in heterogeneous substrates.

Br J Dermatol, 2002 Mar, 146(3), 365 - 74
Slow release iodine preparation and wound healing: in vitro effects consistent with lack of in vivo toxicity in human chronic wounds; Zhou LH et al.; BACKGROUND: Antiseptic agents, particularly slow-release preparations, are increasingly being used in the management of chronic wounds . One such agent, cadexomer iodine, carries iodine (0.9% weight/weight) immobilized in beads of dextrin and epichlorhydrin and has been demonstrated to be highly effective in promoting healing of exudative wounds . However, there have been no studies directly assessing the potential lack of toxicity of cadexomer iodine on human cutaneous tissues . OBJECTIVES: To determine if, within a certain concentration range, cadexomer iodine is non-toxic to human cells and cutaneous tissue and to assess histologically human chronic exudative wounds that are being treated with cadexomer iodine . METHODS: We examined the effects of varying concentrations of cadexomer iodine on the viability of human fibroblasts in culture (by trypan blue exclusion) . The morphology, cellular proliferation capacity (measured by {3H}thymidine uptake), ability to produce alpha 1(I) procollagen chain mRNA, and cell outgrowth from neonatal foreskin explants were also evaluated in human fibroblasts after incubation with various concentrations of cadexomer iodine . Moreover, biopsies of chronic exudative wounds concurrently treated with cadexomer iodine were stained with haematoxylin and eosin or a Gram stain and evaluated microscopically . RESULTS: At concentrations of up to 0.45%, cadexomer iodine was found to be non-toxic to fibroblasts in vitro; there were no changes in viability, morphology, cellular proliferation, ability to produce collagen, and cell outgrowth from explants . In vivo, skin biopsies of chronic exudative wounds being treated with cadexomer iodine demonstrated no evidence of cell necrosis, displayed re-epithelialization, and revealed bacteria within the cadexomer beads . CONCLUSIONS: These studies demonstrate that cadexomer iodine has definite non-toxic concentration ranges for fibroblasts in vitro, which are consistent with a lack of cellular toxicity in human chronic exudative wounds treated with cadexomer iodine . Cadexomer iodine may also have the additional property of trapping microorganisms.

Am Surg, 2002 Apr, 68(4), 390 - 1
Empyema complicating muscle-sparing thoracotomy: the role of wound management; Sees DW et al.; The fascial layers bordering the latissimus dorsi and anchoring the serratus muscles often do not lend themselves to impervious closure during muscle-sparing thoracotomy . Fluid from the subcutaneous space may therefore drain into the pleural cavity after such procedures . If this fluid is contaminated with microorganisms the potential for development of empyema is present . Two patients are presented in whom this scenario was presumed to have occurred . Early intervention in the second patient was felt to have avoided the development of a major empyema.

Environ Toxicol Chem, 2002 Apr, 21(4), 715 - 9
Kinetics of polychlorinated biphenyl dechlorination by Hudson River, New York, USA, sediment microorganisms; Cho YC et al.; The kinetics of polychlorinated biphenyl (PCB) dechlorination by Hudson River (New York, USA) sediment microorganisms were investigated using Aroclor 1242 at 10 concentrations ranging from 0 to 900 ppm (0-11.2 micromol Cl/g sediment) . The time course of PCB dechlorination and population growth were determined by congener-specific analysis and the most-probable-number technique, respectively, over a 44-week incubation period . Dechlorination rate (nmol Cl removed/g sediment/d) was a linear function of PCB concentrations similar to the dechlorination of Aroclor 1248 by sediment microorganisms from the St . Lawrence River (New York, USA) . However, the rate was much slower, with the linear slope being only 24% that of the St . Lawrence River . The threshold concentration below which no dechlorination occurs was (mean +/- standard deviation) 1.06 +/- 0.18 micromol Cl/g sediment (85 +/- 14 ppm), threefold higher than that for the dechlorination of Aroclor 1248 . The maximum extent of dechlorination was greater at higher Aroclor concentrations . Dechlorinating microorganisms did not show any significant growth until late in the lag phase of dechlorination, and their maximum was greater at higher initial Aroclor 1242 concentrations . Although dechlorination rates were significantly lower with the Hudson River inoculum, when normalized to the maximum number of dechlorinating organisms, they were not significantly different from those for Aroclor 1248 by St . Lawrence River microorganisms . These results further support the idea that PCB dechlorination is tightly linked to the growth of dechlorinating microorganisms.

Intern Med J, 2002 Apr, 32(4), 170 - 8
Surface-active phospholipid: a Pandora's box of clinical applications . Part I . The lung and air spaces; Hills BA; Almost everywhere in the body there are phospholipids, not only comprising the lipid bilayer of membranes, but also in the free state . What is seldom appreciated, except in respirology, is that these 'free' phospholipids are unusual in that many are highly surface active . Surface activity is a property of certain substances (surfactants), conferred by their molecular constitution and configuration, which predisposes them to locate at interfaces because, in doing so, they reduce interfacial energy . When adsorbed (reversibly bound) to solid surfaces, surfactants can impart many highly desirable properties that have been widely studied and long accepted in the physical sciences, while their commercial applications have withstood the test of time . These desirable properties include lubricity (boundary lubrication), release (antistick) and dewatering, while providing a barrier to corrosion, abrasion, solute transmission and to biological microorganisms . Many of these offer obvious roles for surface-active phospholipid (SAPL), ranging from a corrosion inhibitor in the stomach to a load-bearing lubricant in the joints . This opens a veritable 'Pandora's box' of potential clinical applications . Part I of this review challenges traditional beliefs in respirology that 'surfactant' is unique to the lung and, moreover, that its actions are confined to the liquid-air interface . Evidence is discussed that, by binding to alveolar epithelium, SAPL imparts semi-permeability needed before channels pumping ions can also pump water vital for maintaining fluid balance . Evidence is also reviewed for a lining to upper airways, sinuses and Eustachian tube where it can act like a standard release agent.

Vnitr Lek, 2002 Feb, 48(2), 142 - 6
{Non-specific immune responses in patients with chronic diabetic foot syndrome and chronic bacterial infection}; Jirkovska A et al.; Chronic bacterial ulcers infection is a frequent cause of non-healing diabetic foot . The major factors of a non-specific immune response are phagocytic cells including polymporphonuclear (PMN) leukocytes, and humoral systems (complement) . PMN leukocytes remove microorganisms by phagocytosis a part of it is intracellular killing and degradation in a process requiring energy and associated with "respiratory burst" . The aim of our study was to assess non-specific immune response in patients with diabetic foot syndrome and chronic bacterial infection . 30 patients treated over one month with antibiotics for an infected diabetic foot in our foot clinic had significantly lower values of "oxidative burst" of PMN leukocytes in basal state (396 +/- 228 vs . 574 +/- 337, p < 0.05) in comparison with 25 matched healthy controls . There were no significant differences neither in the count of active phagocyting PMN leukocytes and their initial phagocytic activity nor in the humoral component of non-specific immunity (in circulating immunocomplexes, C3 and C4 components of complement) between both groups . The results of our study show a slightly altered non-specific immune response in patients with diabetic foot syndrome and chronic bacterial infection.

Anal Biochem, 2002 Apr 15, 303(2), 186 - 93
Highly sensitive and specific detection of viable Escherichia coli in drinking water; Min J et al.; A highly sensitive and specific assay method was developed for the detection of viable Escherichia coli as an indicator organism in water, using nucleic acid sequence-based amplification (NASBA) and electrochemiluminescence (ECL) analysis . Viable E . coli were identified via a 200-nt-long target sequence from mRNA (clpB) coding for a heat shock protein . In the detection assay, a heat shock was applied to the cells prior to disruption to induce the synthesis of clpB mRNA and the mRNA was extracted, purified, and finally amplified using NASBA . The amplified mRNA was quantified with an ECL detection system after hybridization with specific DNA probes . Several disruption methods were investigated to maximize total RNA extracted from viable cells . Optimization was also carried out regarding the design of NASBA primer pairs and detection probes, as well as reaction and detection conditions . Finally, the assay was tested regarding sensitivity and specificity . Analysis of samples revealed that as few as 40 E . coli cells/mL can be detected, with no false positive signals resulting from other microorganisms or nonviable E . coli cells . Also, it was shown that a quantification of E . coli cells was possible with our assay method . (C)2002 Elsevier Science (USA)

Chembiochem, 2001 Oct 1, 2(10), 747 - 57
Novel hyperbranched glycomimetics recognized by the human mannose receptor: quinic or shikimic acid derivatives as mannose bioisosteres; Grandjean C et al.; The mannose receptor mediates the internalization of a wide range of molecules or microorganisms in a pattern recognition manner . Therefore, it represents an attractive entry for specific drug, gene, or antigen delivery to macrophages and dendritic cells . In an attempt to design novel effective synthetic mannose receptor ligands, quinic and shikimic acid were selected as putative mannose mimics on the basis of X-ray crystallographic data from the related rat mannose-binding lectin . As the mannose receptor preferentially binds to molecules displaying several sugar residues, fluorescein-labeled cluster quinic and shikimic acid derivatives with valencies of two to eight were synthesized . Their mannose receptor mediated uptake was assayed on monocyte-derived human dendritic cells by cytofluorimetric analysis . Mannose-receptor specificity was further assessed by competitive inhibition assays with mannan, by confocal microscopy analysis, and by expression of the mannose receptor in transfected Cos-1 cells . Constructs derived from both quinic and shikimic acid were efficiently recognized by the mannose receptor with an optimum affinity for the molecules with a valency of four . As a result, commercially available quinic and shikimic acids appear as stable mannose bioisosteres, which should prove valuable tools for specific cell delivery.

J Bacteriol, 2002 May, 184(9), 2420 - 8
Demonstration that fbiC is required by Mycobacterium bovis BCG for coenzyme F(420) and FO biosynthesis; Choi KP et al.; Using the nitroimidazopyran-based antituberculosis drug PA-824 as a selective agent, transposon-generated Mycobacterium bovis strain BCG (M . bovis) mutants that could not make coenzyme F(420) were identified . Four independent mutants that could not make F(420) or the biosynthesis intermediate FO were examined more closely . These mutants contained transposons inserted in the M . bovis homologue of the Mycobacterium tuberculosis gene Rv1173, which we have named fbiC . Complementation of an M . bovis FbiC(-) mutant with fbiC restored the F(420) phenotype . These data demonstrate that fbiC is essential for F(420) production and that FbiC participates in a portion of the F(420) biosynthetic pathway between pyrimidinedione and FO . Homologues of fbiC were found in all 11 microorganisms that have been fully sequenced and that are known to make F(420) . Four of these homologues (all from members of the aerobic actinomycetes) coded for proteins homologous over the entire length of the M . bovis FbiC, but in seven microorganisms two separate genes were found to code for proteins homologous with either the N-terminal or C-terminal portions of the M . bovis FbiC . Histidine-tagged FbiC overexpressed in Escherichia coli produced a fusion protein of the molecular mass predicted from the M . bovis BCG sequence (approximately 95,000 Da), as well as three other histidine-tagged proteins of significantly smaller size, which are thought to be proteolysis products of the FbiC fusion protein.

Mikrobiol Z, 2002 Jan-Feb, 64(1), 82 - 6
{Frequency of isolation of viable tuberculosis bacteria from patients with different forms of lung tuberculosis}; Iavors'ka GV et al.; Tuberculosis mycobacteria isolated from patients with lung tuberculosis have been analyzed to define their viability . The character of bacterial excretions was studied in patients with different clinical forms and detection genesis of tuberculosis process . During 1998-2000 about 228 cases of growth absence after mycobacteria inoculation have been revealed under positive microscopy . When determining viability of tuberculosis mycobacteria, it was shown that in most cases patients with all the forms of the pathologic process, except for the fibrous-cavernysh form, excrete alive microorganisms . The considerably higher amount of alive tuberculosis mycobacteria was distinguished in people who had chronical or relapsed tuberculosis in comparison with newly diagnosed patients.

Mikrobiol Z, 2001 Nov-Dec, 63(6), 71 - 88
{Interaction of microorganisms with solids}; Kurdish IK; The paper generalizes data from literature dedicated to microorganisms interaction with solids . This interaction is widely spread in nature and considerably affects physiological activity of microbial populations . The role of physical, chemical and biological factors in this process is analyzed . It is noted that under certain conditions the hydrophobic forces are the basic factor in this interaction . The role of the cell surface components in this process is discussed.

Int J Biochem Cell Biol, 2002 Jun, 34(6), 645 - 56
Urea-induced unfolding studies of free- and ligand-bound tetrameric ATP-dependent Saccharomyces cerevisiae phosphoenolpyruvate carboxykinase . Influence of quaternary structure on protein conformational stability; Encinas MV et al.; ATP-dependent phosphoenolpyruvate (PEP) carboxykinases are found in plants and microorganisms, and catalyse the reversible formation of PEP, ADP, and CO(2) from oxaloacetate plus ATP . These enzymes vary in quaternary structure although there is significant sequence identity among the proteins isolated from different sources . To help understand the influence of quaternary structure in protein stability, the urea-induced unfolding of free- and substrate-bound tetrameric Saccharomyces cerevisiae PEP carboxykinase is described and compared with the unfolding characteristics of the monomeric Escherichia coli enzyme {Eur . J . Biochem . 255 (1998) 439} . The urea-induced denaturation of S . cerevisiae PEP carboxykinase was studied by monitoring the enzyme activity, intrinsic protein fluorescence, circular dichroism (CD) spectra, and 1-anilino-8-naphthalenesulfonate (ANS) binding . The unfolding profiles were multi-steps, and formation of hydrophobic structures were detected . The data indicate that unfolding and dissociation of the enzyme tetramer are simultaneous events . Ligand binding, most notably PEP in the presence of MnCl(2), conferred a marked protection against urea-induced denaturation . A similar protection effect was found when N-iodoacetyl-N'-(5-sulfo-1-napthyl)ethylene diamine (1,5-I-AEDANS) was covalently bound at Cys(365), within the active site region . Refolding experiments indicated that total recovery of tertiary structure was only obtained from samples previously unfolded to less than 30% . In the presence of substrates, complete refolding was achieved from samples originally denatured up to 50% . The unfolding behaviour of S . cerevisiae PEP carboxykinase was found to be similar to that of E . coli PEP carboxykinase, however all steps take place at lower urea concentrations . These findings show that, at least for monomeric and tetrameric ATP-dependent PEP carboxykinases, quaternary structure does not contribute to protein conformational stability.

FEBS Lett, 2002 Mar 27, 515(1-3), 44 - 50
Identification of differentially expressed genes by cDNA-AFLP technique during heat stress in cowpea nodules; Simoes-Araujo JL et al.; Legume nodules formed by diazotrophic microorganisms are active sites for biological nitrogen fixation (BNF) . In tropical regions, a significant part of N supply for soybean, peanut and bean crops is derived from BNF, which is nevertheless often limited by high temperature stress . In contrast, cowpea nodules are very resistant to high temperatures . To understand the molecular bases of thermotolerance during BNF under heat stress, we have used cDNA-amplified fragment length polymorphism experiments to identify differentially expressed transcripts from cowpea nodules subjected to heat shock treatment . The expression profiles obtained showed approximately 600 bands, 55 up-regulated and nine corresponding to genes repressed by heat stress . Twenty transcript-derived fragments were isolated, cloned and sequenced . The Vigna unguiculata nodule and stress response transcripts present similarities to those that encode low molecular weight heat shock proteins, wound-induced proteins, disease resistance protein, and xylan endohydrolase isoenzyme, as well as different housekeeping genes . The differential expression of 15 genes was confirmed by using Northern blot or reverse Northern hybridization experiments.

Lipids, 2002 Mar, 37(3), 317 - 23
Selective (R)-3-hydroxylation of FA by Stenotrophomonas maltophilia; Weil K et al.; Soil samples were screened for microorganisms selectively transforming FA . One of the isolated strains was identified as the bacterium Stenotrophomonas maltophilia by its phenotypic features and genotypic characterization by sequencing the ribosomal RNA gene . Using linoleic acid as substrate resulted in the formation of two major compounds . After liquid chromatographic isolation and separation, their structures were elucidated by HPLC-tandem MS, GC-MS, and NMR techniques to be 3-hydroxy-Z6-dodecenoic acid and 3-hydroxy-Z5,Z8-tetradecadienoic acid . In additional experiments, other FA, such as a-linolenic, oleic, palmitoleic, myristoleic, and cis-vaccenic acids, were converted to 3-hydroxylated metabolites of shorter chain lengths as well . Determination of the enantiomeric composition revealed highly enriched (R)-hydroxylation (88-98% enantiomeric excess).

Clin Infect Dis, 2002 May 1, 34(9), 1232 - 42 Epub 2002 Apr 02.
The promise of novel technology for the prevention of intravascular device-related bloodstream infection . I . Pathogenesis and short-term devices; Crnich CJ et al.; Intravascular devices (IVDs) are widely used for vascular access but are associated with substantial risk of development of IVD-related bloodstream infection (BSI) . The development of novel technologies, which are based on an understanding of pathogenesis, promises a quantum reduction in IVD-related infections in an era of growing nursing shortages . Infections of short-term IVDs (that is, those in place <10 days), including peripheral venous catheters, noncuffed and nontunneled central venous catheters (CVCs), and arterial catheters, derive mainly from microorganisms colonizing the skin around the insertion site, which most often gain access extraluminally . More-effective cutaneous antiseptics, such as chlorhexidine, a chlorhexidine-impregnated sponge dressing, CVCs with an anti-infective coating, anti-infective CVC hubs, and novel needleless connectors, have all been shown to reduce the risk of IVD-related BSI in prospective randomized trials . The challenge for the future will be to identify new preventative technologies and to begin to adapt more widely those technologies already shown to be efficacious and cost-effective.

Planta, 2002 Apr, 214(6), 821 - 8 Epub 2001 Dec 14.
Evidence that hydroxyl radicals mediate auxin-induced extension growth; Schopfer P et al.; Reactive oxygen intermediates, i.e . the superoxide radical (O*-)(2), hydrogen peroxide (H2O2) and the hydroxyl radical (*OH), are generally regarded as harmful products of oxygenic metabolism causing cell damage in plants, animals and microorganisms . However, oxygen radical chemistry may also play a useful role in polymer breakdown leading to wall loosening during extension growth of plant cells controlled by the phytohormone auxin . Backbone cleavage of cell wall polysaccharides can be accomplished in vitro by (*OH) produced from H2O2 in a Fenton reaction or in a reaction catalyzed by peroxidase supplied with O2 and NADH . Here, we show that coleoptile growth of maize seedlings is accompanied by the release of reactive oxygen intermediates in the cell wall . Auxin promotes release of (O*-)(2) and subsequent generation of (*OH)when inducing elongation growth . Experimental generation of (*OH) in the wall causes an increase in wall extensibility in vitro and replaces auxin in inducing growth . Auxin-induced growth can be inhibited by scavengers of (O*-)(2), H2O2 or (*OH), or inhibitors interfering with the formation of these molecules in the cell wall . These results provide the experimental background for a novel hypothesis on the mechanism of plant cell growth in which (*OH), produced from (O*-)(2) and H2O2 by cell wall peroxidase, acts as a wall-loosening agent.

Anal Bioanal Chem, 2002 Mar, 372(5-6), 737 - 9 Epub 2002 Feb 16.
A BOD biosensor based on a microorganism immobilized on an Al2O3 sol-gel matrix; Chen D et al.; A novel BOD biosensor has been fabricated from a microbial membrane on an oxygen electrode; a porous inorganic Al2O3 sol-gel matrix was used to immobilize the yeast . Use of the sensor is convenient and rapid compared with the official BOD5 method . The gelation time required to form the microorganism membrane was less than 15 min . There was linear relationship between the response (sensor current) and BOD values ranging from 10-50 mg x L(-1) . The lifetime of the BOD biosensor was more than 30 days at ambient temperature.

J Ind Microbiol Biotechnol, 2002 Jan, 28(1), 42 - 7
Industrial microbiology of solar salt production; Javor BJ; Solar salterns can be modeled as giant outdoor chemostats, much like a series of dams on a slow-moving river . Microorganisms and their products play an essential, but sometimes uncharacterized, role in salt production in these ponds, from seawater salinity up through NaCl saturation . They may physically affect the evaporation process and their by-products may chemically modify or bind with dissolved ions . Many solar salt facilities engage microbiologists to establish monitoring programs for analyses of nutrients, standing crop and associated biological variables in the ponds . Other solar salt companies engage microbiologists only when there are "crises" in the ponds that interfere with salt production.

Ann Biol Clin (Paris), 2002 Mar-Apr, 60(2), 193 - 200
{Emerging parasitosis ans mycosis: risk and threats for the new millenium}; Pinel C et al.; For multiple reasons, the emergent infectious risks do not stop increasing these last twenty years . The climatic modifications and the human interventions modifying the biotope as well as the rapid spreading of resistant strains to treatments, generate re-emergence or emergence, all the more dramatic as the means of fight are reduced . These emergent or re-emergent diseases are extremely worrisome as their diagnosis and their prevention are often difficult . The important infesting power of parasites and the particularly effective capacities of adaptation of these eucaryotes contributed to the public health problems . Anthropozoonoses and zoonoses constitute a permanent risk the control of which is imaginary . The new pathogenic agents, the unusual clinical demonstrations in the context of deficiency of the host immune functions imply attentiveness and a permanent up to date of the knowledge of the biologist and of the different professionals of health . The risks with which are confronted the humanity during this century underline the necessity of determining mechanisms involved in the pathogenesis . The determination of the specific and vital biologic processes for the microorganism, could allow to define the most appropriated targets and the most effective and original means of fight.

Structure (Camb), 2002 Apr, 10(4), 473 - 82
Early structural rearrangements in the photocycle of an integral membrane sensory receptor; Edman K et al.; Sensory rhodopsins are the primary receptors of vision in animals and phototaxis in microorganisms . Light triggers the rapid isomerization of a buried retinal chromophore, which the protein both accommodates and amplifies into the larger structural rearrangements required for signaling . We trapped an early intermediate of the photocycle of sensory rhodopsin II from Natronobacterium pharaonis (pSRII) in 3D crystals and determined its X-ray structure to 2.3 A resolution . The observed structural rearrangements were localized near the retinal chromophore, with a key water molecule becoming disordered and the retinal's beta-ionone ring undergoing a prominent movement . Comparison with the early structural rearrangements of bacteriorhodopsin illustrates how modifications in the retinal binding pocket of pSRII allow subtle differences in the early relaxation of photoisomerized retinal.

Eur Respir J, 2002 Mar, 19(3), 546 - 56
Role of persistent infection in the control and severity of asthma: focus on Chlamydia pneumoniae; von HL; Conventional risk factors have been unable to explain most of the substantial increase in the prevalence of asthma observed in many countries during the last few decades . Much attention has been directed at the "hygiene hypothesis", the apparent inverse relationship between intense systemic childhood infections and the subsequent development of asthma and atopy . However, it is not only the absence or scarcity of infections, but the prolonged presence of certain microorganisms in the lungs that may be involved in the development of asthma . Accumulating evidence suggests that Chlamydia pneumoniae, an intracellular ubiquitous pathogen with an innate propensity to persist and cause chronic infections, may be associated with asthma . This microorganism can achieve a state of "latency" in which it is viable but dormant and does not multiply . During this state, however, chlamydia continues to synthesize the "stress" protein, a 60-kDa heat shock protein (hsp60) . This protein is able to elicit a strong host inflammatory response at sites of its production and appears to be involved in tissue injury and scarring processes . As inflammation has been found to be present in almost all asthmatics, whatever the severity and aetiology of the disease, inhaled glucocorticoids now have an established position in the treatment of early stages . However, corticosteroids negatively affect many aspects of cell-mediated immunity and favour the shift from a T-helper-1-type response towards a T-helper-2-type response . Corticosteroids may thus severely deteriorate the host's ability to eradicate an intracellular pathogen, such as Chlamydia pneumoniae, which requires a properly functioning cell-mediated (T-helper-1-type) immune response to be cleared . These drugs are also able to reactivate persistent Chlamydia to an active growth phase, which, by increasing the production of pro-inflammatory cytokines at the site of infection, can further amplify inflammation in the airways of patients with asthma.

Int Immunol, 2002 Apr, 14(4), 411 - 9
IFN-alpha/beta enhances BCR-dependent B cell responses; Braun D et al.; Type I interferon (IFN-I) is constitutively produced in the bone marrow (BM), and induced at sites of inflammation and following infection by viruses or microorganisms . We have previously shown that IFN-I regulates the generation and selection of normal B cell populations in the BM . In the present work, we assess the effects of IFN-I on mature B cell function by monitoring the responses of IFN-alpha/beta-treated murine splenic B cells to apoptotic, mitogenic and activating stimuli . A similar analysis is performed on BM mature B cells obtained from wild-type or IFN-I receptor-deficient mice . IFN-alpha/beta is shown to induce B cells to a state of partial activation characterized by the up-regulation of CD69, CD86 and CD25 molecules in the absence of either proliferation or terminal differentiation . B cells treated with IFN-alpha/beta show an increased survival and resistance to Fas-mediated apoptosis . IFN-alpha/beta also enhances B cell responses to BCR ligation such as calcium fluxes, IgM internalization, induction of activation markers and proliferation . These results indicate that in addition to its inhibitory effect on viral replication and T cell apoptosis, IFN-alpha/beta plays an essential role during an inflammatory response by lowering the threshold for B cell induction, thereby promoting fast and polyclonal antibody responses.

Am J Respir Crit Care Med, 2002 Apr 1, 165(7), 954 - 60
Occupational asthma in greenhouse flower and ornamental plant growers; Monso E et al.; The objective of this study was to determine the prevalence of occupational asthma and sensitization to workplace allergens in greenhouse flower and/or ornamental plant growers . A random sample of 40 growers who cultivated such crops and had participated in the European Farmers' Study was selected for a cross-sectional study that included (1) greenhouse characteristics and air contaminants (dust, endotoxin, and microorganisms), (2) respiratory symptoms, (3) occupational asthma diagnosed by bronchial provocation challenge, and (4) sensitization to workplace flowers or molds . Associations among respiratory symptoms, sensitization, greenhouse characteristics, and air contaminants were analyzed by logistic regression modeling . Thirty-nine growers agreed to participate (response rate, 97.5%) . Bronchial provocation challenge confirmed occupational asthma in three workers (7.7%), all of them sensitized to workplace flowers or molds . No cases of occupational asthma were found among nonsensitized growers . Poor ventilation proved to be a marginal risk factor for wheezing (air velocity: odds ratio, 0.11; 95% confidence interval, 0.01-1.04) . Sensitization to flowers or molds was found in 13 of 38 growers (34.2%) but was not related to greenhouse characteristics . We conclude that one-fourth of greenhouse flower and/or ornamental plant growers sensitized to workplace flowers or molds have occupational asthma, a disease that is suffered by 8% of the growers who cultivate these crops.

Crit Rev Food Sci Nutr, 2002 Mar, 42(2), 151 - 61
Microbial control by packaging: a review; Cutter CN; Since early man first used a variety of natural containers to store and eat foods, significant developments in food packaging materials have provided the means to suppress microbial growth as well as protect foods from external microbial contamination . Throughout this progression, packaging materials have been developed specifically to prevent the deterioration of foods resulting from exposure to air, moisture, or pH changes associated with the food or the surrounding atmosphere . Both flexible and rigid packaging materials, alone or in combination with other preservation methods, have been developed to offer the necessary barrier, inactivation, and containment properties required for successful food packaging . Examples of flexible packaging used to inactivate microorganisms associated with foods include controlled atmosphere, vacuum, modified atmosphere, active, and edible packaging . Additionally, the combination of rigid packaging materials made from metal, glass, or plastic with heat provides the most effective and widely used method for inactivating microorganisms . As with all food products, it is necessary to integrate a HACCP-based program to assure quality throughout the packaging operation . In addition to packaging improvements, other novel technologies include the development of detectors for oxygen levels, bacterial toxins, and microbial growth, or the integration of time-temperature indicators for detection of improper handling or storage.

Int J Food Microbiol, 2002 Mar, 73(2-3), 395 - 407
Predictive microbiology: towards the interface and beyond; McMeekin TA et al.; This review considers the concept and history of predictive microbiology and explores aspects of the modelling process including kinetic and probability modelling approaches . The "journey" traces the route from reproducible responses observed under close to optimal conditions for growth, through recognition and description of the increased variability in responses as conditions become progressively less favourable for growth, to defining combinations of factors at which growth ceases (the growth/no growth interface) . Death kinetics patterns are presented which form a basis on which to begin the development of nonthermal death models . This will require incorporation of phenotypic, adaptive responses and may be influenced by factors such as the sequence in which environmental constraints are applied . A recurrent theme is that probability (stochastic) approaches are required to complement or replace kinetic models as the growth/no growth interface is approached and microorganisms adopt a survival rather than growth mode . Attention is also drawn to the interfaces of predictive microbiology with microbial physiology, information technology and food safety initiatives such as HACCP and risk assessment.

Int J Food Microbiol, 2002 Mar, 73(2-3), 343 - 9
Seafood Spoilage Predictor--development and distribution of a product specific application software; Dalgaard P et al.; To allow shelf-life prediction of a range of products, the Seafood Spoilage Predictor (SSP) software has been developed to include both kinetic models for growth of specific spoilage microorganisms and empirical relative rates of spoilage models . SSP can read and evaluate temperature profile data of different formats and in this way the software is a flexible device for electronic time-temperature integration . Predicted values of microbial growth and of remaining product shelf life can be exported from SSP as graphs and tables in ASCII, HTML and eXtensible Mark-up Language (XML) formats and this allows SSP to be used in combination with other programmes . More than 300 people have downloaded SSP and distribution of this software from the internet has been efficient in stimulating the application of predictive microbiology and of mathematical seafood shelf-life models within industry, research, seafood inspection and teaching.

Rev Stomatol Chir Maxillofac, 2002 Feb, 103(1), 26 - 9
{Systemic infections of dental origin}; Gilon Y et al.; Microorganisms harbored in the oral cavity have long been recognized to cause systemic disease with a well known mechanism of spread via the blood stream . Different factors, including the presence of periodontal disease, the number of dental extractions or the duration of dental surgery have an influence on the risk of bacteremia . Infectious endocarditis is classically cited, but there are other potential problems including brain abscess, meningitis, lung or lie abscess, and even for some authors, a risk of thrombotic heart disease . It is important to keep in mind that some of these focal infections may be possible complications of dental (or buccal) infection . Prevention is crucial.

Microbes Infect, 2002 Apr, 4(4), 433 - 40
Alternative food-preservation technologies: efficacy and mechanisms; Lado BH et al.; High-pressure processing, ionizing radiation, pulsed electric field and ultraviolet radiation are emerging preservation technologies designed to produce safe food, while maintaining its nutritional and sensory qualities . A sigmoid inactivation pattern is observed in most kinetic studies . Damage to cell membranes, enzymes or DNA is the most commonly cited cause of death of microorganisms by alternative preservation technologies.

Microbes Infect, 2002 Apr, 4(4), 413 - 23
Ecological factors influencing survival and growth of human pathogens on raw fruits and vegetables; Beuchat LR; Outbreaks of human infections associated with consumption of raw fruits and vegetables have occurred with increased frequency during the past decade . Factors contributing to this increase may include changes in agronomic and processing practices, an increase in per capita consumption of raw or minimally processed fruits and vegetables, increased international trade and distribution, and an increase in the number of immuno-compromised consumers . A general lack of efficacy of sanitizers in removing or killing pathogens on raw fruits and vegetables has been attributed, in part, to their inaccessibility to locations within structures and tissues that may harbor pathogens . Understanding the ecology of pathogens and naturally occurring microorganisms is essential before interventions for elimination or control of growth can be devised.

Int J Syst Evol Microbiol, 2002 Mar, 52(Pt 2), 671 - 8
Thiobaca trueperi gen . nov., sp . nov., a phototrophic purple sulfur bacterium isolated from freshwater lake sediment; Rees GN et al.; Two strains of a novel species of phototrophic micro-organism were isolated from the sediments of a shallow, freshwater, eutrophic lake . Both strains grew photolithoheterotrophically with sulfide as an electron donor, transiently accumulating intracellular sulfur globules . Photolithoautotrophic growth was not observed . One strain was designated BCH(T) (the type strain) and was studied in most detail . Cells contained bacteriochlorophyll a, and the dominant carotenoid was lycopene . Cell suspensions were brown . The photosynthetic membranes had a vesicular arrangement . Acetate, propionate, pyruvate, succinate and fumarate were each used as electron donors and carbon sources in the presence of sulfide and bicarbonate . In the presence of light, growth did not occur with hydrogen, thiosulfate or iron(II) . The optimum temperature for growth was between 25 and 30 degrees C, the maximum being 36 degrees C . The G+C content of the genomic DNA of strain BCH(T) was 63 mol% . Analysis of the 16S RNA genes showed that both strains belonged to the gamma-subclass of the Proteobacteria but were phylogenetically distinct from any described phototrophic organisms within the Chromatiaceae . On the basis of phylogenetic and physiological differences from other phototrophic microorganisms, strain BCH(T) is described as a novel species of a new genus, Thiobaca trueperi gen . nov., sp . nov.

Acta Microbiol Pol, 2001, 50(3-4), 189 - 204
Rep-PCR - a variant to RAPD or an independent technique of bacteria genotyping? A comparison of the typing properties of rep-PCR with other recognised methods of genotyping of microorganisms; Baldy-Chudzik K; The paper presents technical aspects of rep-PCR fingerprinting technique and compares its typing abilities, differentiation power and reproducibility with other recognised and recommended genotyping methods . Although rep-PCR fingerprinting is similar to MAAP techniques, it demonstrates some essentially different elements . The data presented in this review, indicate a rep-PCR genomic fingerprinting technique as a highly discriminating, independent screening method for determining the taxonomical diversity of bacterial population.

Z Gastroenterol, 2002 Apr, 40 Suppl 1, S92 - S4
{Extra-intestinal Manifestations of chronic inflammatory Bowel diseases}; Pierer M et al.; Approximately one third of all patients with Crohn's disease and ulcerative colitis suffer from extra-intestinal manifestations of their inflammatory bowel disease . Most commonly those symptoms occur simultaneously with the CED symptoms; they can, however, either precede them or appear later on in the course of the disease . The most frequent extra-intestinal symptoms are arthralgias of peripheral joints and spine, which are usually bland and self-limiting, while ankylosing spondylitis and erosive arthritides are rare.Skin lesion and eye affections can also parallel the bowel condition, but occasionally they precede intestinal manifestations and can be a first diagnostic clue . In addition, different extra-intestinal symptoms tend to simultaneously co-occur more frequently in some patients with CED, while others are not affected at all.Immunogenetic parameters play a role for the manifestations of the pathologic immune response both in the gut and in the musculo-skeletal systems, as indicated by associations with MHC class I alleles . Enteropathic microorganisms are also thought to be involved in the pathogenetic mechanisms.

Mol Microbiol, 2002 Feb, 43(3), 703 - 15
An ABC-type, high-affinity urea permease identified in cyanobacteria; Valladares A et al.; Urea is an important nitrogen source for many microorganisms, but urea active transporters have not been characterized at a molecular level in any bacterium . Cells of Synechocystis sp . PCC 6803 and Anabaena sp . PCC 7120 exhibited the capacity to take up {14C}-urea from low-concentration (<1 microM) urea solutions . The Ks of Anabaena cells for urea was about 0.11 microM, and the observed uptake activity involved the transport and metabolism of urea . In contrast to urease, which was constitutively ex-pressed, expression of the high-affinity urea uptake activity was subjected to nitrogen control . In an Anabaena ureG (urease-) mutant, a concentrative, active transport of urea could be demonstrated . We found that a mutant of open reading frame (ORF) sll0374 from the Synechocystis genomic sequence lacked urea transport activity . This ORF encoded a conserved component of an ABC-type transporter, but it is not clustered together with any other possible transporter-encoding gene . An Anabaena homologue of sll0374, urtE, was isolated and found to be part of a cluster of genes, urtABCDE, putatively encoding all the elements of an ABC-type permease . Although the longest transcript that we could detect only covered urtABC, the impairment of urea transport by inactivation of urtA, urtB or urtE suggested that the whole gene cluster is expressed producing the urea permease . Expression was induced under nitrogen-limiting conditions, and a complex promoter regulated by the cyanobacterial global nitrogen control transcription factor NtcA was found upstream from urtA . Our work adds urea to the known substrates of the versatile class of ABC-type transporters and suggests the involvement of a transporter of this superfamily in urea scavenging by some bacteria in natural environments.

J Agric Food Chem, 2002 Apr 10, 50(8), 2256 - 60
Resistance of soybean vegetative storage proteins (S-VSPs) to proteolysis by rumen microorganisms; Guenoune D et al.; Soybean vegetative storage proteins (S-VSPs) are lysine-rich and, hence, are potentially of high nutritive value for high productive ruminants . Using S-VSPs from wild-type soybean and from transgenic tobacco plants expressing either one of the two S-VSPs subunits (S-VSP alpha or S-VSP beta) or both, we tested their stability in cow rumen fluid under in situ conditions, using SDS-polyacrylamide gel electrophoresis . Proteolysis and degradation pattern of S-VSPs from transgenic tobacco leaves occurred relatively fast compared with that of wild-type (WT) soybean plants . Comparing the two S-VSPs subunits expressed in transgenic plants, we found that S-VSP alpha was degraded much faster than S-VSP beta . The degradation pattern of S-VSPs in transgenic tobacco plants expressing both subunits resembled that of WT soybean . In contrast, the degradation pattern of transgenic tobacco plants expressing a single subunit was different . These finding suggest that the quaternary structure of S-VSPs may be an important factor determining their resistance to rumen degradation . Our results also suggest that the stability to rumen proteolysis of a given protein, when expressed in a transgenic plant, may not always be predictable and has to be verified.

Mol Ecol, 2002 Mar, 11(3), 571 - 81
Community analysis of arbuscular mycorrhizal fungi associated with Ammophila arenaria in Dutch coastal sand dunes; Kowalchuk GA et al.; A polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) approach for the detection and characterization of arbuscular mycorrhizal fungi (AMF) 18S ribosomal DNA (rDNA) was developed and applied to the study of AMF communities associated with the main sand-stabilizing plant species of the Dutch sand dunes, marram grass (Ammophila arenaria, L.) . DNA was extracted directly from plant roots, soil or isolated AMF spores, and prominent bands resulting from AMF-specific DGGE profiles were excised for sequence analysis . This strategy provided a robust means of detecting and identifying AMF-like species without the use of trap plant cultivation methods . A number of Glomus-like and Scutellospora-like sequences was detected, including a putatively novel Glomus species, and differences were observed in the dominant AMF-like populations detected in healthy vs . degenerating stands of A . arenaria and in bulk sand dune soil . It has previously been suggested that plant pathogens, such as fungi and nematodes, may contribute to the decline of A . arenaria . Although no causal relationship can be drawn between the observed differences in the dominantly detected AMF-like populations and the vitality of plant growth, these results indicate that mutualistic interactions between this plant and AMF should not be overlooked when examining the role of soil-borne microorganisms in vegetation dynamics . In addition, there were discrepancies observed between the AMF-like groups detected in spore populations vs . direct 18S rDNA analysis of root material, corroborating previous suggestions that spore inspection alone may poorly represent actual AMF population structure.

J Biosci, 2002 Feb, 27(1 Suppl 1), 27 - 33
Horizontal gene transfer and bacterial diversity; Dutta C et al.; Bacterial genomes are extremely dynamic and mosaic in nature . A substantial amount of genetic information is inserted into or deleted from such genomes through the process of horizontal transfer . Through the introduction of novel physiological traits from distantly related organisms, horizontal gene transfer often causes drastic changes in the ecological and pathogenic character of bacterial species and thereby promotes microbial diversification and speciation . This review discusses how the recent influx of complete chromosomal sequences of various microorganisms has allowed for a quantitative assessment of the scope, rate and impact of horizontally transmitted information on microbial evolution.

Ophthalmology, 2002 Apr, 109(4), 687 - 93
Rapid direct antibiotic susceptibility testing in endophthalmitis; Mino de Kaspar H et al.; OBJECTIVE: To compare a new rapid antibiotic susceptibility test (RAST) to the conventional method in patients with endophthalmitis . DESIGN: Prospective nonrandomized comparative trial . PARTICIPANTS: Intraocular aspirates from 24 consecutive patients with endophthalmitis were tested . METHODS: Approximately 0.25 ml of vitreous or anterior chamber aspirate was obtained and tested for antibiotic sensitivity using the Kirby-Bauer agar disk diffusion method . Using this conventional testing method, the aspirates were cultured for bacterial growth before antibiotic susceptibility testing (AST) . In contrast, for direct RAST, the aspirates were inoculated directly onto agar plates with antibiotic disks without first culturing for bacterial growth . RESULTS: Of the 24 endophthalmitis aspirates studied, 4 (17%) could not be tested by direct RAST because either more than one bacterial species was present or no bacteria were cultured . The remaining 20 cases provided 467 antibiotic-microorganism combinations . The two methods yielded identical susceptibility results in 409 (88%) of the 467 tests . In 42 tests (9%), a minor discrepancy occurred between the conventional method and RAST, in which one resulted in intermediate susceptibility, whereas the other was either resistant or sensitive . Significant discrepancy, defined as a bacterial sample reported as sensitive in one method yet resistant in the other, occurred with 16 samples (3%) . Results of the RAST were available within 6 to 10 hours compared with more than 24 hours for the conventional method . Of clinical significance, the results of the RAST revealed that in 11 cases of epidemic Stenotrophomonas maltophilia endophthalmitis, the bacteria were resistant to the antibiotics prescribed, and change of antibiotic therapy was made on the basis of the RAST results . CONCLUSIONS: RAST provided accurate antibiotic susceptibility results in a much shorter time than did the conventional method . We suggest that conventional AST be performed for confirmation of the RAST results, but initial antibiotic therapy can quite reliably be based on the results of RAST.

EXS, 2002, (92), 247 - 303
Reexamining microbial evolution through the lens of horizontal transfer; Planet PJ; Our ability to understand the evolution of microbial organisms revolves around a central and increasingly unsettled question: what is the nature of the mode of inheritance? The extent to which genetic information is passed vertically from parent to daughter or horizontally between distant relatives must guide reconstructions and inferences of evolutionary history, and has direct bearing on any ideas about the mechanisms of selection and diversification . Recent evidence suggests that we may have previously underestimated the contribution of horizontal gene transfer, and the dynamics and extent of this process are only beginning to be understood . The recent flood of complete genome sequences of microorganisms has already presented us with a vast array of data from which to test our hypotheses about the evolution of the entire tree of life, but what remains unclear is how we can make sense of this unwieldy data set . Analyses of this newly available data set should include explicit examinations of the contributions of both types of inheritance.

Ned Tijdschr Tandheelkd, 1997 Aug, 104(8), 300 - 1
{Rational use of antibiotics by the dentist can prevent microbial resistance}; van Kasteren ME et al.; Worldwide resistance of microorganisms to antibiotics is becoming a problem of major concern . An antibiotic policy which tries to control the development of resistance by rational use is needed in all fields where antibiotics are prescribed . In dental practice, apart from endocarditisprophylaxis, prophylaxis is leukopenic patients and prophylaxis after dental implants, antibiotics can be used to support local therapy of an dentoalveolar abscess or refractory periodontitis . In the majority of these cases, a small spectrum penicillin is therapy of choice.

Adv Exp Med Biol, 2002, 504, 53 - 69
Biological control of Fusarium head blight of wheat and deoxynivalenol levels in grain via use of microbial antagonists; Schisler DA et al.; Efforts to reduce mycotoxin contamination in food logically start with minimizing plant infection by mycotoxin producing pathogens . Fusarium graminearum (perfect state, Gibberella zeae) infects wheat heads at flowering, causing the disease Fusarium head blight (FHB) and losses of over 2.6 billion dollars in the U.S . during the last 10 years . The pathogen often produces deoxynivalenol (DON) resulting in grain size and quality reduction . Highly resistant wheat cultivars currently are not available for reducing FHB, and labeled fungicides are not consistently effective . The feasibility of biologically controlling FHB is currently being evaluated . Microbial isolates obtained from wheat anthers were screened for their ability to utilize tartaric acid, a compound that is poorly utilized by F . graminearum and could be utilized in formulations of biological control agents . Four strains that utilized tartaric acid and three that did not were effective in reducing FHB disease severity by up to 95% in greenhouse and 56% in field trials . Additional research programs around the globe have identified other antagonist strains with potential for biologically controlling FHB . Though a considerable body of research remains to be completed, strategies and microorganisms for biologically controlling FHB have reached an advanced stage of development and offer the promise of being an effective tool that could soon contribute to the reduction of FHB severity and DON contamination of grain in commercial agriculture.

Gastrointest Endosc, 2002 Apr, 55(4), 518 - 22
Duration of antibiotic therapy for cholangitis after successful endoscopic drainage of the biliary tract; van Lent AU et al.; BACKGROUND: Drainage of the obstructed biliary tree is the mainstay of therapy for patients with acute cholangitis; antibiotic therapy is complementary . It is unknown whether it is necessary to continue therapy with antibiotics once biliary drainage is achieved and signs of systemic inflammation have subsided . METHODS: Patients who presented with acute cholangitis and were successfully treated at ERCP were studied retrospectively . Patients were followed for 6 months after ERCP . RESULTS: Eighty patients fulfilled study criteria . In 46% of patients blood cultures grew microorganisms . All patients recovered from the episode under study . Antibiotic therapy after ERCP was given for a median duration of 3 days (range: 0-42) . Forty-one patients received antibiotic therapy for 3 days or less, 19 for 4 or 5 days, and 20 patients longer than 5 days . The 3 groups were well-matched . In none of the patients did the index episode of cholangitis result in a secondary complication not present at the time of ERCP . The percentage of patients with recurrent cholangitis (24%) was not statistically different for the 3 groups (p = 0.80) . CONCLUSIONS: Short-duration antibiotic therapy (3 days) appears sufficient when adequate drainage is achieved and fever is abating.

Crit Rev Oncol Hematol, 2002 Apr, 42(1), 25 - 34
Carcinogenicity, mutagenicity and teratogenicity of manganese compounds; Gerber GB et al.; Manganese, an essential trace element, is one of the most used metals in the industry . Recently, several new manganese compounds have been introduced as fungicide, as antiknock agent in petrol and as contrasting agent in nuclear magnetic resonance tomography . Manganese displays a somewhat unique behaviour with regard to its toxicity . It is relatively non-toxic to the adult organism except to the brain where it causes Parkinson-like symptoms when inhaled even at moderate amounts over longer periods of time . Relatively high doses of manganese affect DNA replication and repair in bacteria and causes mutations in microorganism and mammalian cells although the Ames test does not appear to be particularly responsive to manganese . In mammalian cells, manganese causes DNA damage and chromosome aberrations . Information on organic manganese derivatives is still insufficient . Large amounts of manganese affect fertility in mammals and are toxic to the embryo and foetus . The fungicide MANEB and the contrasting agent MnDPDP also can be embryotoxic, but the latter only at doses much higher than those clinically employed . Information on the anti-knock agent MMT is inadequate . On the other hand, manganese deficiency can also affect fertility and be teratogenic . Information on cancer due to manganese is scanty but the results available do not indicate that inorganic manganese is carcinogenic . More information is desirable with regard to the organic manganese derivatives . It may surprise that an agent that causes mutations is not also carcinogenic . The experience with manganese shows that conclusions with regard to carcinogenicity of an agent based on the observation of mutations are subject to uncertainties . Altogether, it appears that, because of the very high doses at which positive effects have been found, manganese would not represent a significant carcinogenic risk to the population and workers . Care must, however, be exercised with respect to central-nervous symptoms after chronic exposure and with respect to effects on the embryo . Pregnant women should not be exposed to manganese at the work place.

Clin Chim Acta, 2002 May 7, 319(1), 9 - 17
Is the neutrophil reactive oxygen species production measured by luminol and lucigenin chemiluminescence intra or extracellular? Comparison with DCFH-DA flow cytometry and cytochrome c reduction; Caldefie-Chezet F et al.; BACKGROUND: Polymorphonuclear neutrophils (PMNs) are crucial in host defense against invading microorganisms through reactive oxygen species (ROS) production . However, generated ROS released in excess into media can damage the host tissue . It is therefore essential, when exploring oxygen species production, to discriminate between its intracellular (IC) and extracellular (EC) localization . Several methods of ROS detection are commonly used . However, the literature shows that it is not always clear whether the species detected are IC or EC, especially with the chemiluminescence technique . METHODS: We compared PMN ROS production, determined by chemiluminescence, using two different probes (luminol and lucigenin) with that measured by 2'-7'-dichlorofluorescin diacetate (DCFH-DA) flow cytometry for IC production and by cytochrome c reduction for EC production . RESULTS: We found that luminol-dependent chemiluminescence explored IC ROS production more specifically (r=0.77, p<0.01: correlation between luminol-amplified chemiluminescence and DCFH-DA flow cytometry) . Lucigenin-amplified chemiluminescence and cytochrome c reduction were closely related (r=0.55, p<0.01) . CONCLUSION: Luminometry detection can thus afford reproducible information on intracellular ROS kinetic production using luminol and extracellular ROS detection using lucigenin, simply and at low cost.

J Comp Physiol {B}, 2002 Apr, 172(3), 191 - 6 Epub 2002 Jan 10.
Properties of Na(+)-dependent nucleoside transport in the proximal and distal small intestine of cows; Scharrer E et al.; Large amounts of nucleic acids associated with rumen microorganisms are digested in the proximal part of the small intestine of ruminants . We studied how the proximal-distal gradient in nucleic acid digestion is related to activity of Na(+)-nucleoside transporters in brush border membrane vesicles isolated from the proximal and distal small intestine of cows . Two Na(+)-dependent nucleoside transporters with overlapping substrate specificity were shown to be present at the two intestinal sites, one for pyrimidine nucleosides and one for purine nucleosides . Affinity constants (K(m)-values) for both thymidine and guanosine transport were similar at the two intestinal sites, while transport capacity (V(max)) was 2-3 times higher in the proximal than in the distal small intestine . Glucose and alpha-methyl-D-glucoside (0.1 mmol/l or 2 mmol/l) inhibited transport of thymidine and guanosine markedly only in the proximal small intestine . It is concluded that absorption of nucleosides by the two Na(+)-nucleoside transporters reflects the proximal-distal gradient in nucleic acid digestion.

Infect Control Hosp Epidemiol, 2002 Mar, 23(3), 147 - 51
Scissors: a potential source of nosocomial infection; Embil JM et al.; This point-prevalence survey of healthcare workers' scissors demonstrates that of 232 scissors sampled, 182 (78.4%) were colonized with bacteria . The scissors of nurses and those for communal use were most frequently contaminated . Cleaning of scissors occurred infrequently, but wiping scissors with an alcohol swab effectively disinfected them . The suspicion that scissors may harbor and potentially transmit microorganisms, including antibiotic-resistant bacteria, has been confirmed.

Cornea, 2002 Apr, 21(3), 275 - 83
Amniotic membrane graft for the management of scleral melting and corneal perforation in recalcitrant infectious scleral and corneoscleral ulcers; Ma DH et al.; PURPOSE: To study the efficacy and safety of using cryopreserved human amniotic membrane (AM) graft as a patch graft to reduce stromal melting and promote reepithelialization in extensive infectious scleral and corneoscleral ulcers . METHODS: Four cases of infectious scleral ulcers with persistent scleral melting and no sign of reepithelialization and three cases of corneoscleral ulcers with corneal perforation were studied . All patients had previously undergone pterygium excision, and infections were caused by Pseudomonas (n = 4), fungi (n = 2), and atypical Mycobacterium (n = 1) . The area of limbus involved ranged from 3 to 9 (mean, 4.7) o'clock positions . Repeated debridements were performed, the causative microorganisms were identified, and the appropriate topical and systemic antibiotics were given to all patients before AM grafting . Postoperatively, the speed of reepithelialization, changes in the severity of scleral melting and inflammation, recurrence of infection, and visual acuity were documented . RESULTS: Melting and inflammation at the lesion site decreased after AM grafting . Reepithelialization of the scleral lesions was complete at an average 15.7 +/- 8.7 days (range, 5-31) postoperatively . Focal melting of the AM graft occurred in two cases, and in one case, it was necessary to perform further corneoscleral graft . No recurrent infection was encountered, but sterile abscess occurred in three cases that were located away from the original lesion . Useful vision above 20/400 was maintained in all patients at the end of follow-up . CONCLUSION: The AM graft is effective in promoting conjunctival reepithelialization and reducing scleral melting and inflammation and can be considered as an alternative biomaterial to improve wound healing in scleral and corneoscleral ulcerations.

Ann Nutr Metab, 2002, 46(1), 38 - 44
Chronic ethanol feeding affects intestinal mucus lipid composition and glycosylation in rats; Kaur J; Effect of chronic ethanol feeding on mucus lipid composition and glycosylation was investigated in rat intestine . Ethanol feeding to rats daily for 40 days enhanced cholesterol, phospholipids, triglyceride and cerebroside contents of intestinal mucus . Ethanol feeding exhibited a differential effect on various sugar contents of the mucus . Total hexose content was increased (42%) whereas hexosamines, sialic acid and fucose contents were reduced (p < 0.001) . Results on incorporation of labeled sugars into mucus revealed an enhanced (p < 0.01) galactosylation and reduced (p < 0.01) glucosamination of glycoproteins . Separation of glycoproteins on SDS-PAGE showed that proteins of molecular weight >200, 118, 65 and 55 kD were affected . These were highly galactosylated and less glucosaminated in ethanol-fed animals compared to controls . Thus, ethanol feeding markedly alters the lipid composition and glycosylation process of intestinal mucus in rats . These alterations may be of pathological significance in view of the role of mucus in protection and adhesion of microorganisms .

Ann Nutr Metab, 2002, 46(1), 9 - 16
Effect of heat-treated proteins on selected parameters of the biotransformation system in the rat; Wenzel E et al.; The intake of heat-damaged proteins from food causes various effects, like the loss of essential amino acids and a reduced protein digestibility . There is also an influence on gastrointestinal microorganisms and different digestion enzymes . Until now, very little is known about the influence of heat-treated proteins on the enzymes of the biotransformation system . In the present study, the influence of protein-bound L-lysino-D,L-alanine, N(epsilon)-fructoselysine, and N(epsilon)-carboxymethyllysine (CML) on selected enzymes of the biotransformation in liver, kidney, and intestinal mucosa of male Wistar rats was examined . The contents of cytochrome P-450 and cytochrome b(5) and the activity of NADPH-cytochrome c reductase served as indicators of phase I biotransformation . The influence on phase II biotransformation was shown by the content of glutathione and the glutathione S-transferase activity . The results showed that treatment with heat-damaged proteins mainly affected phase II biotransformation enzymes with CML, yielding the strongest effect . The activity of glutathione S-transferase in the kidney was 86% higher in animals treated with diets containing 4,930 mg.kg(-1) protein-bound CML than in animals of the control group which received a diet without any detectable CML . In addition, a higher level of glutathione was found in the kidneys of animals fed on diets containing CML . The glutathione S-transferase activity was 64% higher in the intestinal mucosa of animals fed on protein-bound N(epsilon)-fructoselysine (2,700 mg.kg(-1)) . The glutathione S-transferase activity was higher (p >0.05) in the intestinal mucosa of animals fed on protein-bound L-lysino-D,L-alanine (2,582 and 12,474 mg.kg(-1)) . In conclusion, ingestion of heat-treated proteins led to an activation of the enzymes of phase II biotransformation . Whether or not the released pure compounds or the degradation products of the test proteins are responsible for the altered enzyme activities remains to be evaluated .

Trends Microbiol, 2002 Apr, 10(4), 193 - 9
Intracellular vs extracellular recognition of pathogens--common concepts in mammals and flies; Girardin SE et al.; There are common themes in innate immune defense systems across the animal and plant kingdoms . Pathogen recognition is commonly based on the identification of microbial molecular patterns by defined receptors and the subsequent activation of signaling pathways that initiate a defense response to fend off the invading microorganism . The existence of mammalian Toll-like receptors (TLRs) and the recent identification of two mammalian nucleotide-binding site leucine-rich repeat (NBS-LRR) proteins (NOD1 and NOD2) as intracellular sensors of bacterial products bring new insights into the possibility of extracellular versus intracellular pathogen recognition and signal transduction depending on the nature of the infection . The homology between TLRs and the Toll system in Drosophila suggests that conserved defense mechanisms are likely to be shared by diverse organisms.

J Biotechnol, 2002 May 9, 95(2), 99 - 108
Carbon and nitrogen substrates consumption, ammonia release and proton transfer in relation with growth of Geotrichum candidum and Penicillium camemberti on a solid medium; Aldarf M et al.; It is difficult to determine fungal biomass growing on a solid medium directly . Therefore, indirect ways of measuring this growth have to be used instead . For this purpose, the kinetics of growth, of substrate consumption and of release of metabolites must be correlated . This has been carried out for Geotrichum candidum and Penicillium camemberti growing on a gelified lactic acid+peptone medium . Peptone was shown to be preferred by G . candidum as a carbon and energy source, in contrast to the behaviour observed for P . camemberti . At the end of growth, G . candidum released about three times more ammonia than P . camemberti . In both cases, the total number of protons consumed during growth was close to the sum of the number of moles of metabolized lactic acid and formed ammonia . During the main part of growth, constant biomass on substrate yields (peptone, lactic acid) was observed for both microorganisms . Therefore, in pure solid cultures, the biomass concentration of both species could be deduced either from DeltapH or from the reduction of substrates . From the above, an estimate of G . candidum biomass in mixed culture could be derived from the determination of ammonia release.

Ceska Slov Farm, 2002 Jan, 51(1), 6 - 10
{Penicillin G acylase--synthesis, regulation, production}; Grafkova J et al.; Penicillin G acylase (PGA) is one of very important industrial enzymes used in the production of polysynthetic beta-lactam antibiotics . This enzyme catalyzes the hydrolysis of the amidic bond of penicillin G with the development of 6-aminopenicillanic acid which serves as the initial substance for the production of semisynthetic penicillins . In the strain Escherichia coli W ATCC 11105 and ATCC 9637, PGA is coded by the pga gene on the chromozome and synthesized as the pre-pro-PGA (pp PGA) precursor, which is transported, with probable participation of the chaperon system, to the periplasmatic space of the cell . Here after a series of proteolytic reactions the active enzyme PGA develops, consisting of two subunits alpha and beta . Expression of the pga gene is subject to several regulatory mechanisms: temperature repression, catabolic repression by glucose, repression by oxygen, and induction by phenylacetic acid (FOK) . The formation of active PGA is also influenced at the post-translation level, where an important role is played by intracellular proteolytic reactions and the transport system of pre-pro-PGA across the cytoplasmatic membrane . The chromozomal area of the pga gene of the E . coli W strain was employed for the construction of many recombinant plasmids . These plasmids served to transform suitable host strains, some of which are now used in industry as highly productive microorganisms.

Biochim Biophys Acta, 2002 Feb 13, 1589(1), 1 - 13
Toll-like receptors as adjuvant receptors; Kaisho T et al.; The mammalian Toll-like receptors (TLRs) are expressed on macrophages and dendritic cells, which are primarily involved in innate immunity . At present, ligands for several of the TLRs, such as TLR2, TLR3, TLR4, TLR5, TLR6, and TLR9, have been identified . Most of these ligands are derived from pathogens, but not found in the host, suggesting that the TLRs are critical to sensing invading microorganisms . Pathogen recognition by TLRs provokes rapid activation of innate immunity by inducing production of proinflammatory cytokines and upregulation of costimulatory molecules . Activated innate immunity subsequently leads to effective adaptive immunity . In this regard, the TLRs are considered to be adjuvant receptors . Distinct TLRs can exert distinct, but overlapping sets of biological effects . Accumulating evidence indicates that this can be attributed to both the common and unique aspects of the signaling mechanisms that mediate TLR family responses . For example, TLR2 and TLR9 require MyD88 as an essential signal transducer, whereas TLR4 can induce costimulatory molecule upregulation in a MyD88-independent manner . Understanding the TLR system should offer invaluable opportunity for manipulating host immune responses.

Aviat Space Environ Med, 2002 Mar, 73(3), 211 - 5
Cabin air quality: an overview; Rayman RB; In recent years, there have been increasing complaints from cockpit crew, cabin crew, and passengers that the cabin air quality of commercial aircraft is deficient . A myriad of complaints including headache, fatigue, fever, and respiratory difficulties among many others have been registered, particularly by flight attendants on long-haul routes . There is also much concern today regarding the transmission of contagious disease inflight, particularly tuberculosis . The unanswered question is whether these complaints are really due to poor cabin air quality or to other factors inherent intlight such as lowered barometric pressure, hypoxia, low humidity, circadian dysynchrony, work/rest cycles, vibration, etc . This paper will review some aspects relevant to cabin air quality such as volatile organic compounds (VOCs), carbon dioxide (CO2), carbon monoxide (CO), ozone (O3), particulates, and microorganisms, as well as the cabin ventilation system, to discern possible causes and effects of illness contracted inflight . The paper will conclude with recommendations on how the issue of cabin air quality may be resolved.

Arch Microbiol, 2002 Mar, 177(3), 259 - 66 Epub 2002 Jan 23.
Morphological and physiological changes in Streptomyces lividans induced by different yeasts; Santamaria RI et al.; Streptomyces development is a complex process that eventually finishes with the formation of individual unigenomic spores from the aerial hyphae . Intraspecific and interspecific signals must play a key role in triggering or blocking this process . Here we show that interaction between two types of microorganisms, Streptomyces and yeasts, leads to alteration of the Streptomyces developmental program . This alteration is due to the action of invertase produced by the yeast on the sucrose present in the culture media, making glucose and fructose readily available for growth.

Lancet Oncol, 2001 Apr, 2(4), 221 - 5
Marine organisms as a source of new anticancer agents; Schwartsmann G et al.; Various active anticancer agents are derived from plants and terrestrial microorganisms . The isolation of C-nucleosides from the Caribbean sponge, Cryptotheca crypta, four decades ago, provided the basis for the synthesis of cytarabine, the first marine-derived anticancer agent to be developed for clinical use . Cytarabine is currently used in the routine treatment of patients with leukaemia and lymphoma . Gemcitabine, one of its fluorinated derivatives, has also been approved for use in patients with pancreatic, breast, bladder, and non-small-cell lung cancer . Over the past decade, several new experimental anticancer agents derived from marine sources have entered preclinical and clinical trials . This field has expanded significantly as a result of improvements in the technology of deep-sea collection, extraction, and large-scale production through aquaculture and synthesis . In this paper, examples of marine-derived experimental agents that are currently undergoing preclinical and early clinical evaluation are briefly discussed . A summary of the available information on the results of phase I and II trials of agents such as aplidine, ecteinascidin-734 (ET-734), dolastatin 10 and bryostatin 1 is also presented.

Nat Prod Rep, 2002 Feb, 19(1), 1 - 48
Marine natural products; Faulkner DJ; This review covers the marine natural products literature for the year 2000 and is organized phylogenetically, with sections on marine microorganisms and phytoplankton, green algae, brown algae, red algae, sponges, coelenterates, bryozoans, molluscs, tunicates . echinoderms and miscellaneous marine organisms . There is an emphasis on new structures, stressing their biological activities, source organisms and countries of origin, and also syntheses that confirm the structures of known compounds . The review contains 869 structures and 592 references, of which 434 appeared between January and December 2000.

Curr Gastroenterol Rep, 2002 Apr, 4(2), 167 - 71
The role of microorganisms in biliary tract disease; Ljungh A et al.; The biliary tract is normally sterile, but bile-tolerant bacteria are frequently isolated from patients with cholecystitis . Since the identification of about 25 Helicobacter species, some of which may grow in bile, studies have addressed the role of these organisms in primary biliary cirrhosis, primary sclerosing cholangitis, and cholelithiasis . Most of these bacteria show the presence of Helicobacter DNA or antigens in the bile tract and in liver samples . Altogether, data from studies on biliary and hepatic diseases, as well as pancreatic disorders, suggest that bile-tolerant Helicobacter species may induce a chronic infection with possible malignant transformation.

Biosci Rep, 2001 Aug, 21(4), 419 - 44
Keratitis; Sharma S; Corneal inflammation or keratitis is a significant cause of ocular morbidity around the world . Fortunately, the majority of the cases are successfully managed with medical therapy, but the failure of therapy does occur, leading to devastating consequences of either losing the vision or the eye . This review attempts to provide current information on most, though not all, aspects of keratitis . Corneal inflammation may be ulcerative or nonulcerative and may arise because of infectious or noninfectious causes . The nonulcerative corneal inflammation may be confined to the epithelial layer or to the stroma of the cornea or may affect both . For clarity, this section has been divided into nonulcerative superficial keratitis and nonulcerative stromal keratitis . While the former usually includes hypersensitivity responses to microbial toxins and unknown agents, the latter can be either infectious or noninfectious . In the pathogenesis of ulcerative keratitis, microorganisms such as bacteria, fungi, parasites (Acanthamoeba), or viruses play an important role . Approximately, 12.2% of all corneal transplantations are done for active infectious keratitis . Available world literature pertaining to the incidence of microbial keratitis has been provided special place in this review . On the other hand, noninfectious ulcerative keratitis can be related to a variety of systemic or local causes, predominantly of autoimmune origin.

Ann Transplant, 2001, 6(3), 6 - 9
Current status and perspectives for the generation of transgenic pigs for xenotransplantation; Niemann H; Xenotransplantation implies transplantation of organs between discordant, e.g . non-related species . This procedure usually is associated with a hyperacute rejection response (HAR) which destroys the transplanted organ within minutes . To overcome the growing shortage of human organs, transgenic pigs have been generated that express human complement regulatory genes . This approach enables to overcome the HAR as shown by an extended average survival rate of 40-90 days of the immunosuppressed primate recipient of a transgenic porcine heart . It is expected that transgenic pigs will be available as organ donors within the next 5-7 years . A major prerequisite is the prevention of the potential transfer of pathogenic microorganisms, in particular porcine endogenous retroviruses (PERV) . Transgenic livestock has been generated predominantly via microinjection of DNA-constructs into pronuclei of zygotes . However, efficiency is low and only 1-3% transgenic offspring are to be obtained . Integration of the transgene occurs at random and expression is independent from the number of integrated copies but can be affected by the integration site . Improvements of the efficiency in the generation of transgenic pigs will be achieved by the use of genetically modified donor cells in nuclear transfer technology (cloning).

Folia Microbiol (Praha), 2001, 46(5), 417 - 22
Vegetative growth, aging- and light-induced conidiation of Trichoderma viride cultivated on different carbon sources; Chovanec P et al.; The growth and conidiation of the aged Trichoderma viride culture grown in the dark, and after an induction by a light pulse, was examined in the presence of selected mono-, di(tri)saccharides, amino acids and alcohols as sole carbon sources . Hexoses and disaccharides, but not pentoses and amino acids, promoted proportionally both growth and conidiation induced by aging or light . All compounds but pentoses promoted the conidiation in aged cultures and photoconidiation in a close correlation . Ethanol, glycerol and ethylene glycol supported both growth and conidiation but these processes were not supported equally . Conidia formation with hexoses and amino acids as sole carbon sources seems to be a function of growth promotion, rather than of growth restriction (starvation, stress, aging) . With glucose as sole carbon source the conidiation was not triggered by nutrient limitation, nor by the accumulation of waste metabolites . The aging-induced conidiation can be considered to be triggered by the genetic program of the microorganism rather than by its nutrient status.

Curr Mol Med, 2001 Nov, 1(5), 589 - 95
Receptors for unopsonized particles: the role of alveolar macrophage scavenger receptors; Palecanda A et al.; The lung is constantly exposed to potentially pathogenic particles and microorganisms . Alveolar macrophage (AM) binding of inhaled environmental particles is a critical first step in phagocytosis and clearance, and must be accomplished without the benefit of opsonization by specific antibodies . Opsonin-independent phagocytosis is initiated by direct recognition of phagocytic target . The identities of receptors on AMs that mediate unopsonized particle binding were, until recently, not known . Using flow cytometry, monoclonal antibody and expression cloning techniques we have found a major role for the scavenger receptor, MARCO in AM binding of particles and bacteria . In this review we will discuss the role of scavenger receptors in AM binding of unopsonized particles and the use of flow cytomety in analyzing AM-particle interaction . We will also discuss other non-scavenger receptors involved in opsonin-independent phagocytosis.

Curr Mol Med, 2001 Sep, 1(4), 437 - 46
Biology of gammadelta T cells in tuberculosis and malaria; Dieli F et al.; Tuberculosis and malaria remain the leading causes of mortality among human infectious diseases in the world . It is estimated that 3 to 5 million people die from tuberculosis and malaria each year . Although it is traditionally believed that CD4 and CD8 alphabeta T lymphocytes are mandatory for protective immune responses against Mycobacterium tuberculosis and Plasmodium falciparum (the ethiologic agents of tuberculosis and the most severe form of malaria, respectively), there is still incomplete understanding of the mechanisms of immune protection and of the causes of its failure in the affected patients . Several studies in humans and animal models have suggested that Vgamma9/Vdelta2 T cells may play an important role in the immune responses against Mycobacterium tuberculosis and Plasmodium falciparum . Vgamma9/Vdelta2 T cells represent about 75% of all circulating gammadelta T cells while they can be greatly expanded during the acute phase of Mycobacterium tuberculosis and Plasmodium falciparum malaria . Vgamma9/Vdelta2 T recognize a new class of antigenic molecules which are nonpeptidic in nature and contain critical phosphate moieties (phosphoantigens) . Interestingly, phosphoantigens isolated from Mycobacterium tuberculosis and Plasmodium falciparum share strong structural homology and are probably identical . However, despite a large body of data reported in the literature, it is not yet clear whether Vgamma9/Vdelta2 T cells play a protective or pathogenic role in immune responses against Mycobacterium tuberculosis and Plasmodium falciparum . In this review we summarize our current knowledge of the biology of Vgamma9/Vdelta2 T cells in response to the two pathogens, Mycobacterium tuberculosis and Plasmodium falciparum, and provide evidence suggesting definition of a novel and important protective role through which Vgamma9/Vdelta2 T cells can contribute to the killing of microorganisms residing in intracellular compartments.

Curr Mol Med, 2001 Jul, 1(3), 339 - 55
Mechanisms of T cell receptor antagonism: implications in the treatment of disease; Dittel BN; The adaptive immune response is often required for the successful clearing of infectious pathogens . Antigen presenting cells (APC) present peptide antigens derived from pathogens to T cells via major histocompatibility complex (MHC) molecules . T cells then become activated and differentiate into effector cells with the capacity to kill infected cells or to induce an anti-pathogen antibody response . In autoimmunity, this T cell response is directed against self-antigens and often leads to deleterious effects on specific tissues . Likewise, T cell responses to allogeneic MHC molecules in transplants also leads to pathology . By introducing subtle changes in the antigenic peptide amino acid content, T cell activation can be inhibited, thereby preventing T cell effector functions . This strategy of TCR antagonism has been used successfully in vitro and in vivo to inhibit models of autoimmunity and allorecognition . In addition, a variety of pathogens that often result in chronic disease following infection, also have seemingly evolved natural mechanisms to inhibit T cell responses by antagonism . These microorganisms express natural variants of certain proteins, that when presented to T cells have the capacity to specifically inhibit T cell responses by functioning as antagonists or by modulating the nature of the T cell response . The understanding of how pathogens mediate this inhibition in vivo will be beneficial to ongoing studies in both autoimmunity and transplantation aimed at suppressing the harmful immune response, thereby controlling disease . TCR antagonism seems to have the potential to be used therapeutically to prevent or inhibit an undesired T cell response that will ultimately lead to disease.

J Food Prot, 2002 Mar, 65(3), 571 - 4
Reduction of spoilage microorganisms in fresh beef using hydrodynamic pressure processing; Williams-Campbell AM et al.; Hydrodynamic pressure processing (HDP) was investigated as a technology to reduce spoilage microorganisms found in fresh beef . In two separate studies (studies 1 and 2), retail ground beef and beef roasts were purchased (day 0) . The roasts were divided into stew pieces (30 to 40 g) . All meat samples, including control samples, were stored at 5 degrees C for 20 h in a plastic film . After storage, designated samples were treated with HDP In study 3, ground beef was treated with HDP (day 0) and stored aerobically (5 degrees C) for 14 days with control samples . Each meat type was vacuum-packaged for HDP (100 g binary explosive, steel shock wave container) . The pHs and the aerobic plate counts (log10 CFU/g) were measured on day 0 (studies I and 2) and on days 0, 7, and 14 (study 3) for control samples and for HDP-treated samples . There was no pH difference between control and HDP-treated meat types (studies 1 and 2); HDP reduced bacteria in both meat types in study 1 (2 log) and study 2 (1.5 log) on day 0 . In study 3, there was a significant difference (P < 0.05) in pH between control meat (8.2) and HDP-treated meat (5.6) after storage . There was an immediate reduction (1.5 log) of microorganisms following HDP (day 0) and a 4.5-log difference between control samples (9 log) and HDP-treated samples (4.5) after 14 days of storage . With HDP, it is possible to reduce spoilage microorganisms found in or on different meat types (ground beef versus stew pieces), which could extend the shelf life of meat products.

Folia Microbiol (Praha), 2001, 46(6), 495 - 503
Morphological details of microorganisms revealed by RCH-microscopy at high magnification--a ready-to-use adaptation of a light microscope; Zizka Z et al.; RCH-microscopy (Relief Contrast after Hostounsky) is a new method of optical microscopy in transmitted light developed with Lambda Ltd., Prague . This method was used to study bacteria, fungi including yeasts and algae at high magnification . The equipment provides a three-dimensional image of high contrast and resolution . The results of these microscopic observations can be used for both morphological (taxonomical) and ecological studies of microorganisms.

N Y State Dent J, 2002 Feb, 68(2), 34 - 7
W . D . Miller . The pioneer who laid the foundation for modern dental research; Ring ME; Toothache has been the most ubiquitous ailment to plague mankind from time immemorial . Until the late 1700s, it was thought that the cause of this torment was the wriggling, in a carious tooth, of a worm . And early attempts at treatment were focused on driving the "worm" out . It was one of the world's greatest scientists, the dentist W . D . Miller, who, after extensive research, in 1891 published his epochal work, The Microorganisms of the Human Mouth, which set forth a new theory regarding the cause of dental caries . His postulating a "chemico-parasitic" origin of caries laid the basis for all the modern research in dentistry aimed at wiping this scourge out . Contemporary research has proven the worth of Miller's groundbreaking theory.

Science, 2002 Mar 15, 295(5562), 2067 - 70
Metabolic activity of subsurface life in deep-sea sediments; D'Hondt S et al.; Global maps of sulfate and methane in marine sediments reveal two provinces of subsurface metabolic activity: a sulfate-rich open-ocean province, and an ocean-margin province where sulfate is limited to shallow sediments . Methane is produced in both regions but is abundant only in sulfate-depleted sediments . Metabolic activity is greatest in narrow zones of sulfate-reducing methane oxidation along ocean margins . The metabolic rates of subseafloor life are orders of magnitude lower than those of life on Earth's surface . Most microorganisms in subseafloor sediments are either inactive or adapted for extraordinarily low metabolic activity.

Infect Immun, 2002 Apr, 70(4), 1816 - 23
Entamoeba histolytica-induced dephosphorylation in host cells; Teixeira JE et al.; Activation of host cell protein tyrosine phosphatases (PTPases) and protein dephosphorylation is an important mechanism used by various microorganisms to deactivate or kill host defense cells . To determine whether protein tyrosine dephosphorylation played a role in signaling pathways affecting Entamoeba histolytica-mediated host cell killing, we investigated the involvement of PTPases during the attachment of E . histolytica to target cells . We observed a rapid decrease in cellular protein tyrosine levels in Jurkat cells, as measured with an antiphosphotyrosine monoclonal antibody, following adherence to E . histolytica . Ameba-induced protein dephosphorylation was contact dependent and required intact parasite, since blocking amebic adherence with galactose inhibited tyrosine dephosphorylation and amebic lysates had no effect on phosphotyrosine levels . Moreover, disruption of amebic adherence with galactose promoted recovery of phosphorylation in Jurkat cells, indicating that dephosphorylation precedes target cell death . The evidence suggests that ameba-induced dephosphorylation is mediated by host cell phosphatases . Prior treatment of Jurkat cells with phenylarsine oxide, a PTPase inhibitor, inhibited ameba-induced dephosphorylation . We also found proteolytic cleavage of the PTPase 1B (PTP1B) in Jurkat cells after contact with amebae . The calcium-dependent protease calpain is responsible for PTP1B cleavage and enzymatic activation . Pretreatment of Jurkat cells with calpeptin, a calpain inhibitor, blocked PTP1B cleavage and inhibited ameba-induced dephosphorylation . In addition, inhibition of Jurkat cell PTPases with phenylarsine oxide blocked Jurkat cell apoptosis induced by E . histolytica . These results suggest that E . histolytica-mediated host cell death occurs by a mechanism that involves PTPase activation.

Allergy Asthma Proc, 2002 Jan-Feb, 23(1), 53 - 7
Common variable immunodeficiency; Abonia JP et al.; We present a case report and review of the literature that illustrates many key features of patients with common variable immunodeficiency (CVID) . These patients frequently present with repeated infections with a variety of different microorganisms . Recurrent sinopulmonary infections can lead to serious chronic complications such as bronchiectasis, and gastrointestinal infections can result in malabsorption . In addition to serious infection, CVID is associated with a number of comorbid disorders including a variety of autoimmune diseases and neoplasms . Here, we provide an illustrative case report and discuss the primary features and therapy for patients with CVID.

Environ Monit Assess, 2002 Feb, 74(1), 45 - 65
Changes in soil properties of abandoned shrimp ponds in southern Thailand; Towatana P et al.; Chemical soil properties between active shrimp ponds and abandoned ones on the Bangkok soil series were compared, at Ranote District, Songkhla Province in southern Thailand . Soil samples were collected at depth intervals of 0-10, 10-20, 20-30, 30-40 and 40-50 cm from pond bottoms at the same ponds used in a former study conducted in 1994, for a total of 6 ponds with 3 sampling sites for each pond . These ponds were active during the previous study in 1994, abandoned in 1996 and investigated by this study in 1999 . All the samples were analyzed for exchangeable Ca, Mg, K and Na electrical conductivity (EC), organic matter, S, P and pH, and statistically compared with the analytical results of the previous study . An increase in amounts of Ca, Mg, K, Na and EC in the abandoned ponds as compared with the active ones by 1.3-3.4, 1.4-2.1, 7.0-30.0, 1.2-6.3 and 1.3-10.9 times respectively was observed . That more of these elements were gained than lost each time the seawater was introduced into the ponds, is explained by the 'Element Input/Output Consideration' as proposed herein . Furthermore, a decrease in organic matter, S and P was also observed in the abandoned pond soils, and attributed to the absence of shrimp food and shrimp excreta following the cessation of shrimp raising activities . An unexpected decline in the soil pH of the abandoned ponds was found as well . Aerobic decomposition of organic matter during the absence of shrimp raising activities caused by soil microorganisms triggering SO2 and H2SO4 formation probably played a more significant role than the increase in the amounts of the basic elements (Ca, Mg, K, and Na) eventually reducing soil pH in the abandoned ponds . The significant depletion of the amounts of organic matter in the abandoned pond soils also supports this observation.

J Chemother, 2002 Feb, 14(1), 54 - 8
Penetration of piperacillin/tazobactam (4 g/500 mg) into synovial tissue; Boselli E et al.; The degree of penetration of an antibiotic into the infected site is an important criterion for therapeutic success . This is particularly true for bone and joint infections . The association of piperacillin and tazobactam has been widely used in the treatment of serious infections including bone infections, but no study has been devoted to the subject of its diffusion into synovial tissue . Our objective was to quantify piperacillin/tazobactam synovial tissue penetration and to estimate the efficacy of the association against the microorganisms usually encountered in joint infections . In an open-label study, 6 subjects with similar age, weight, height and creatinine clearance, who were undergoing elective total hip replacement, received a single, parenteral, 4 g/500 mg dose of piperacillin/tazobactam . Plasma and synovial tissue samples were collected and analyzed by a validated HPLC method . The mean concentrations of piperacillin and tazobactam 1.5 h after the initiation of infusion were 69.9 +/- 4.9 microg/mL and 7.7 +/- 0.3 microg/mL, respectively, in plasma and 37.1 +/- 2.1 microg/g and 2.8 +/- 0.4 microg/g, respectively, in synovial tissue . The synovial tissue/plasma ratios were 0.5 +/- 0.0 for piperacillin and 0.4 +/- 0.0 for tazobactam . The piperacillin/tazobactam ratios were 9.1:1 in plasma and 13.5:1 in synovial tissue . The concentrations achieved in synovial tissue are above the MICs of most of the susceptible pathogens usually involved in joint infections, which suggests that the piperacillin/tazobactam combination should be effective in the treatment of most joint infections caused by susceptible microorganisms.

Curr Rheumatol Rep, 2002 Apr, 4(2), 113 - 22
Scleroderma-like cutaneous syndromes; Mori Y et al.; Several distinct entities associated with dermal fibrosis can mimic scleroderma/systemic sclerosis . The list of scleroderma-like conditions or scleroderma variants includes eosinophilic fasciitis, localized forms of scleroderma, scleredema and scleromyxedema, keloids, and environmental exposure-associated conditions including eosinophilia-myalgia syndrome and pseudosclerodermas induced by various drugs . Although these conditions are relatively uncommon, their accurate recognition is essential to avoid misdiagnosis and inappropriate therapy . The pathogenesis of these scleroderma variants appears to share similarities with each other and with that of scleroderma . Better understanding of scleroderma-like disorders is emerging through epidemiologic investigations, and in vivo and in vitro experimental research . Activation of eosinophils and disordered regulation of fibroblast collagen synthesis, apoptosis, and proliferation are recurrent findings in these disorders . The etiologic role of infection with Borrelia species or other microorganisms remains controversial . Cytokines such as transforming growth factor-beta, interleukin-4, interleukin-13, and connective tissue growth factor contribute to fibrosis in these disorders by inducing an accentuated and persistent fibrogenic response to tissue injury . The role of genetic factors in susceptibility and clinical expression of scleroderma-like conditions remains to be systematically addressed . Because of the relative rarity of these conditions, few well-controlled clinical treatment trials have been performed . In addition, there is no consensus on optimal management . Much anecdotal information and small clinical series indicate that phototherapy may have a role in the treatment of scleroderma-like conditions.

APMIS, 2001 Oct, 109(10), 656 - 64
In situ hybridisation for identification and differentiation of Mycoplasma hyopneumoniae, Mycoplasma hyosynoviae and Mycoplasma hyorhinis in formalin-fixed porcine tissue sections; Boye M et al.; Oligonucleotide probes targeting 16S ribosomal RNA were designed for species-specific identification of the porcine mycoplasmas Mycoplasma hyopneumoniae, Mycoplasma hyorhinis and Mycoplasma hyosynoviae using a fluorescent in situ hybridisation assay . The specificity of the probes was evaluated using pure cultures as well as porcine tissue sections with artificial presence of mycoplasma, and the probes were found specific for the target organisms . The assay was applied on sections of 28 tissue samples from pigs infected with one or more of the three Mycoplasma species as determined by cultivation . M . hyopneumoniae and M . hyorhinis were identified in accordance with cultivation in lung sections from nine pigs affected by catarrhal to purulent bronchopneumonia . Likewise, in eight cases of fibrinous pericarditis, M . hyopneumoniae, M . hyorhinis and M . hyosynoviae were the infectious agents according to cultivation and were correctly identified by in situ hybridisation . Out of 11 joints cultivation positive for M . hyosynoviae, the probe was only able to identify M . hyosynoviae in eight cases probably due to a low number of microorganisms in the tissue sections . The in situ hybridisation assay is well suited for use in diagnostic and experimental work as well as a tool for pathogenesis studies.

Toxicol Pathol, 2002 Jan-Feb, 30(1), 54 - 8
Immune responses: adverse versus non-adverse effects; Kimber I et al.; The adaptive immune system in vertebrates has evolved to provide host resistance to infectious microorganisms and malignant disease . Normal immune function and the induction of specific immune responses require the orchestrated interaction between cells and molecules both within and outside the lymphoid system . Immunotoxicology can be defined as the study of adverse health effects that may result from the interaction of xenobiotics with the immune system . In general terms such effects can take one of two forms . The first of these is immunotoxicity (or immunosuppression) where there is a perturbation of, or damage to, one or more components of the immune system resulting in impaired immune function and reduced host resistance . The design and interpretation of experimental immunotoxicity studies and the investigation of clinical immunosuppression require consideration of the relationship between changes in the structure and/or function of discrete components of the immune system and holistic changes in the susceptibility to infectious and malignant disease . The other main way in which chemicals may cause adverse health effects secondary to interaction with the immune system is through stimulation of specific immune responses that result in allergic disease . Allergy to chemicals and proteins can take many forms, including allergic contact dermatitis, allergic sensitization of the respiratory tract (associated with rhinitis and/or asthma), systemic allergic reactions (associated frequently with drug treatment), and gastrointestinal disease . Here there is a need to distinguish between immunogenic responses per se and those immune responses that are of sufficient vigor and of the quality necessary to provoke allergic sensitization . The purpose of this article is to explore the extent to which distinctions can be drawn between adverse and nonadverse effects in the context of immunotoxicity and allergy.

J Anim Sci, 2002 Mar, 80(3), 790 - 6
A comparison of enzymatic and molecular approaches to characterize the cellulolytic microbial ecosystems of the rumen and the cecum; Michalet-Doreau B et al.; We used RNA probes and enzyme activities to compare the cellulolytic microbial ecosystems of the rumen and the cecum . Four rumen- and cecum-cannulated wethers were fed a diet of barley plus hay (60:40) . Digesta samples were collected 1 h before feeding and 3, 6, and 9 h after feeding for measurements on microbial populations, and 1 h before feeding and 3 and 6 h after feeding for digestion measurements, pH, and VFA . Polysaccharidase and glycosidase specific activities of solid-adherent microorganisms were measured respectively by the amount of reducing sugars released from xylan or avicel or p-nitrophenol from the p-nitrophenol derivatives of xylose and glucose . The distribution and amounts of the three main cellulolytic bacterial species (Fibrobacter succinogenes, Ruminococcus albus, and Ruminococcus flavefaciens) were determined by dot-blot hybridization using specific 16SrRNA-targeting probes . Enzyme activities were higher in the rumen than in the cecum and before feeding than at 3 h after feeding . The sum of the three cellulolytic bacterial species represented, on average, 4.5% of the total bacterial RNA in the two compartments and did not vary with sampling time . The cellulolytic bacterial community structure was different in the two compartments, with F . succinogenes as the main species in the rumen and R . flavefaciens in the cecum . The lower cellulolytic activity in the cecum than in the rumen could not be ascribed to any difference in the structure of the cellulolytic bacterial community between these two compartments, and other hypotheses related to digestion are proposed.

Curr Opin Investig Drugs, 2001 Oct, 2(10), 1382 - 6
The role of the gut lymphoid tissue in induction of oral tolerance; Song F et al.; The gut-associated lymphoid tissue (GALT) maintains a balance between immunological tolerance to dietary proteins and induction of active immune responses to pathogenic microorganisms . The oral administration of soluble protein antigens induces a state of systemic immunological unresponsiveness specific to the fed protein, termed oral tolerance . The two major mechanisms to explain oral tolerance are anergy/deletion of autoreactive lymphocytes and active suppression . This review will discuss the mechanisms of therapeutic oral tolerance in relation to events occurring at the site of antigen entry.

Transplantation, 2002 Feb 27, 73(4), 565 - 8
Chronic parvovirus B19 infection in a pediatric lung transplanted patient; Moreux N et al.; In immunocompromised patients, clinical manifestations of human parvovirus B19 (PVB19) infection are mostly reported as acute or chronic hematological disorders . Recently, PVB19 infection has been associated with nonhematological symptoms . Four years after lung transplantation, a 9-year-old girl developed a severe anemia with reticulocytopenia requiring blood transfusion . PVB19 DNA was found by polymerase chain reaction in blood . Blood marrow aspiration revealed typical features of PVB19 infection . She was successfully treated with high dose of i.v . Ig . Then, she exhibited recurrent nonregenerative anemia requiring another course of i.v . Ig . PVB19 DNA has been persisted in blood with no specific immune response . At the same time, she suffered from several lung infection syndromes with no microorganism found except PVB19 DNA . Recurrent mild renal dysfunction was noticed with no other explanation than PVB19 infection . This report indicates that pediatric transplanted patients are at risk of chronic PVB19 infection, which can be associated with lung and/or renal disorders.

EMBO J, 2002 Mar 15, 21(6), 1241 - 7
Yeast vacuoles and membrane fusion pathways; Wickner W; Selective membrane fusion underlies subcellular compartmentation, cell growth, neurotransmission and hormone secretion . Its fundamental mechanisms are conserved among organelles, tissues and organisms . As befits a conserved process, reductionism led to its study in microorganisms . Homotypic fusion of the vacuole of Saccharomyces cerevisiae is particularly accessible to study as vacuoles are readily visualized, there is a rapid and quantitative in vitro assay of vacuole fusion, and the genetics and genomics of this organism and of vacuole fusion are highly advanced . Recent progress is reviewed in the context of general questions in the membrane fusion field.

Arch Biochem Biophys, 2002 Mar 15, 399(2), 232 - 8
Cloning and characterization of an S-formylglutathione hydrolase from Arabidopsis thaliana; Kordic S et al.; A cDNA from Arabidopsis thaliana resembling S-formylglutathione hydrolase (SFGH), an enzyme with putative roles in formaldehyde detoxification in animals and microorganisms, has been cloned and expressed in Escherichia coli . The purified recombinant Arabidopsis enzyme (AtSFGH) was a dimer composed of 31-kDa subunits . Like SFGHs from other sources, AtSFGH had thioesterase activity toward S-formylglutathione and carboxyesterase activity toward 4-methylumbelliferyl acetate . Unlike other SFGHs, the enzyme from Arabidopsis actively hydrolyzed S-acetylglutathione . AtSFGH activity was inhibited by heavy metals and sulfhydryl alkylating agents, but was insensitive to serine hydrolase inhibitors, suggesting that the enzyme was a cysteine-dependent hydrolase . Although Atsfgh transcripts were determined in plants and cultures of Arabidopsis, the respective enzyme could not be detected in planta after the esterase activities present were resolved using isoelectric focusing . Instead, Arabidopsis contained several carboxyesterases active toward alpha-naphthyl acetate, which were all sensitive to inhibition by the serine hydrolase inhibitor paraoxon.

Ann Periodontol, 2001 Dec, 6(1), 9 - 15
The association between periodontal diseases and cardiovascular diseases: a state-of-the-science review; Beck JD et al.; Early case-control and cross-sectional studies demonstrating associations between chronic periodontitis and cardiovascular disease (CVD) were quickly followed by secondary analyses of data available from existing longitudinal studies, which indicated that individuals with periodontitis, as determined by clinical measures, were at greater risk for CVD events . Many of these studies contained large numbers of subjects and were adjusted for traditional risk factors . Within the last 18 months, one case-control study and one longitudinal study have reported finding positive associations that were not statistically significant . The earlier studies stimulated a number of studies focused on identifying potential biological mechanisms that might underlie this association . While still early in that process, such studies have implicated a systemic role for oral microorganisms and for the quality and quantity of the host inflammatory response as key biologic processes that may underlie the association of CVD with the clinical manifestation of periodontitis . It is a positive development when changes in our knowledge regarding biologic mechanisms result in reevaluation of past studies, and this reevaluation leads to new studies that incorporate the design elements demanded by this new knowledge . In that spirit, we conclude that all longitudinal studies reported to date can be characterized as follows: none were initially designed to actually test the association of interest; almost all were restricted to clinical measures of periodontitis to index the exposure and lacked measures of infectious burden and host response; and they used a variety of cardiovascular clinical events to index the outcome and did not include subclinical measures of atherosclerosis . In addition, the longitudinal studies that failed to show a significant association between periodontitis and CVD used the least sensitive and crudest clinical measures of periodontal disease . Based upon the current state-of-the-science, all previous studies should be viewed as lacking sufficiently sensitive and comprehensive measures of periodontal disease as a systemic exposure . Since the potential health care impact of this relationship might be extensive, it is time to enter the next phase of research by conducting molecular epidemiology studies that are appropriately designed to test our current understanding of the molecular and cellular mechanisms involved.

Ann Periodontol, 2001 Dec, 6(1), 138 - 45
Treatment of periodontal disease and control of diabetes: an assessment of the evidence and need for future research; Grossi SG; Evidence points to an increased cytokine response in type 2 diabetes, especially the proinflammatory cytokines interleukin (IL)-1 beta, IL-6, and tumor necrosis factor (TNF)-alpha . Genetics, age, and, nutrition are important signals for this increased response and as reported more recently, infections and inflammation . Persistent elevation of IL-1 beta, IL-6, and TNF-alpha in the diabetic state have an effect on the liver, stimulate the release of acute-phase proteins, produce the characteristic dysregulation of lipid metabolism associated with type 2 diabetes, and have effects on pancreatic beta cells as well . In addition, TNF-alpha, a potent inhibitor of the tyrosine kinase activity of the insulin receptor, has been implicated as an etiologic factor for insulin resistance . Collectively, the evidence supports a role for cytokine elevation in the pathophysiology and metabolic abnormalities associated with diabetes . Periodontitis is an infection that is twice as prevalent in diabetic individuals compared to non-diabetics . Porphyromonas gingivalis, one of the microorganisms responsible for this infection, is able to invade endothelial cells and is a potent signal for monocyte and macrophage activation . Thus, once established in the diabetic host, this chronic infection complicates diabetes control and increases the occurrence and severity of microvascular and macrovascular complications . Unlike treatment of acute infections, modalities of treatment for chronic infections are a matter of debate . Evidence indicates that mechanical removal of subgingival infection does not result in complete elimination of periodontal infection and consequently there is no effect on diabetes control measured as reduction in glycated hemoglobin . On the other hand, studies incorporating systemic antibiotics as adjuncts to mechanical debridement result in a reduction of P . gingivalis to nondetectable levels and a concomitant reduction in glycated hemoglobin, independent of the hypoglycemic effects of diabetes drugs or insulin . The evidence supports the notion that treatment of chronic periodontal infection is essential in the diabetic patient . Assessment of infection status in diabetic patients is fundamental for appropriate treatment decisions.

Bioorg Med Chem, 2002 May, 10(5), 1337 - 46
Substituted indoloquinolines as new antifungal agents; Ablordeppey SY et al.; Cryptolepine (2) possesses desirable properties to serve as a lead in developing new antifungal agents . Using SAR techniques, several analogues of cryptolepine were designed to increase potency and to broaden the antifungal spectrum over several opportunistic microorganisms . A number of 2-substituted indoloquinolines have been synthesized and evaluated in antifungal screens and several have been shown to increase potency and expand the antifungal spectrum of cryptolepine . Comparison of MICs of a number of these analogues with standard antifungal agents, shows them to be comparable to Amphotericin B and Ketoconazole.

Eur J Clin Invest, 2002 Mar, 32 Suppl 1, 84 - 90
Iron chelation and hydroxyl radical scavenging reduce the inflammatory response of endothelial cells after infection with Chlamydia pneumoniae or influenza A; Visseren FL et al.; BACKGROUND: Chronic low-grade inflammation is associated with increased risk of vascular diseases . The source of inflammation is unknown but may well be chronic and/or repetitive infections with microorganisms . Direct infection of endothelial cells (ECs) may also be a starting point for atherogenesis by initiating endothelial procoagulant activity, increased monocyte adherence and increased cytokine production . We hypothesized that iron-mediated intracellular hydroxyl radical formation after infection is a key event in triggering the production of interleukin-6 (IL-6) by ECs in vitro . METHODS: Cultured ECs were incubated with Fe(II) and Fe(III) or infected with Chlamydia pneumoniae or influenza A/H1N1/Taiwan/1/81 for 48 and 24 h, respectively . To determine the role of iron and reactive oxygen species, cells were coincubated with the H2O2 scavenger N-acetyl-l-cysteine, with the iron chelator deferoxamine (DFO) or with the intracellular hydroxyl radical scavenger dimethylthiourea (DMTU) . After the incubation periods, supernatants were harvested for IL-6 determination . RESULTS: Incubating ECs with Fe(II) and Fe(III) resulted in increased IL-6 production . Similarly, infection with C . pneumoniae and influenza A also induced an IL-6 response . Coincubating ECs with DFO or DMTU blocked this response . Nuclear factor-kappaB activity was increased after infection and blocked by coincubation with DFO or DMTU . CONCLUSION: Cultured ECs respond to infection and iron incubation with increased production of IL-6 . Iron, the generation of intracellular hydroxyl radical and NF-kappaB activity are essential in cellular activation, suggesting that reactive oxygen species generated in the Haber-Weiss reaction are essential in invoking an immunological response to infection by ECs.

Environ Sci Pollut Res Int, 2002, 9(1), 29 - 47
Phytoremediation of polyaromatic hydrocarbons, anilines and phenols; Harvey PJ et al.; Phytoremediation technologies based on the combined action of plants and the microbial communities that they support within the rhizosphere hold promise in the remediation of land and waterways contaminated with hydrocarbons but they have not yet been adopted in large-scale remediation strategies . In this review plant and microbial degradative capacities, viewed as a continuum, have been dissected in order to identify where bottle-necks and limitations exist . Phenols, anilines and polyaromatic hydrocarbons (PAHs) were selected as the target classes of molecule for consideration, in part because of their common patterns of distribution, but also because of the urgent need to develop techniques to overcome their toxicity to human health . Depending on the chemical and physical properties of the pollutant, the emerging picture suggests that plants will draw pollutants including PAHs into the plant rhizosphere to varying extents via the transpiration stream . Mycorrhizosphere-bacteria and -fungi may play a crucial role in establishing plants in degraded ecosystems . Within the rhizosphere, microbial degradative activities prevail in order to extract energy and carbon skeletons from the pollutants for microbial cell growth . There has been little systematic analysis of the changing dynamics of pollutant degradation within the rhizosphere; however, the importance of plants in supplying oxygen and nutrients to the rhizosphere via fine roots, and of the beneficial effect of microorganisms on plant root growth is stressed . In addition to their role in supporting rhizospheric degradative activities, plants may possess a limited capacity to transport some of the more mobile pollutants into roots and shoots via fine roots . In those situations where uptake does occur (i.e . only limited microbial activity in the rhizosphere) there is good evidence that the pollutant may be metabolised . However, plant uptake is frequently associated with the inhibition of plant growth and an increasing tendency to oxidant stress . Pollutant tolerance seems to correlate with the ability to deposit large quantities of pollutant metabolites in the 'bound' residue fraction of plant cell walls compared to the vacuole . In this regard, particular attention is paid to the activities of peroxidases, laccases, cytochromes P450, glucosyltransferases and ABC transporters . However, despite the seemingly large diversity of these proteins, direct proof of their participation in the metabolism of industrial aromatic pollutants is surprisingly scarce and little is known about their control in the overall metabolic scheme . Little is known about the bioavailability of bound metabolites; however, there may be a need to prevent their movement into wildlife food chains . In this regard, the application to harvested plants of composting techniques based on the degradative capacity of white-rot fungi merits attention.

Acta Odontol Latinoam, 1996, 9(1), 13 - 9
Germicide effect of several glass ionomer cements; Molgatini SL et al.; One the most significant characteristics of glass ionomer cements is their ability to release fluoride compounds . This study was carried out to try establish relationships between this property and the possible effect on the growth of microorganisms that are found in carious lesions, Agar BHI medium containing Petri dishes were flooded with strains of Actinomyces naeslundii, Actinomyces israelii and Actinomyces odontolyticus . Cavities were then prepared in the agar and filled with mixtures of several glass ionomer cements . Some of them were polymerizable resin containing products . A zinc phosphate and a zinc oxide-eugenol cement were used as controls . After a seven day incubation at 37 degrees C under anaerobic conditions the inhibition halos around the specimens were measured in a way similar to that used for antibiograms . The statistical analysis of the results showed no significant differences among Actinomyces strains but a significant difference one among cements . Even when no definitive conclusions could be drawn it is worth taking into consideration the effect of glass ionomer cements on microorganisms such as the Actinomyces and continuing studies to establish more clearly what is required from the material to produce a clinically significant outcome.

J Immunol, 2002 Mar 15, 168(6), 2626 - 33
Homeostatic regulation of intestinal villous epithelia by B lymphocytes; Nishiyama Y et al.; The epithelial cell of the small intestine is one of the most rapidly regenerating cells in the body . However, the cellular mechanism and biological significance underlying this rapid regeneration remain elusive . In this study we examined the intestinal epithelia of mutant mice that lack B and/or T cells and those of normal littermates . The absence of B cells in Ig mu-chain mutant mice or B and T cells in recombination-activating gene (RAG)-2(-/-) as well as SCID mutant mice was associated with a marked acceleration of epithelial cell turnover and an up-regulation of the expression of MHC class II molecules . No such effects were observed in T cell-deficient TCR-delta and -beta double-mutant mice . As far as the goblet cells of villous epithelium are concerned, absolute numbers of them remained the same among these mutant mice that have no B and/or T cells . Alymphoplasia (aly/aly) mutant mice that lacked Peyer's patches and Ig-producing cells in the lamina propria, but harbored a large number of intestinal mucosal T cells, also displayed a significant acceleration of epithelial cell turnover and, to some extent, up-regulated expression of MHC class II molecules . Notably, the accelerated epithelial cell turnover was not observed and returned to normalcy in the Ig mu-chain mutant mice that had been given antibiotic-containing water . These findings indicate that B cells down-regulate the generation and differentiation of intestinal epithelial cells in the normal wild-type condition and suggest that enteric microorganisms are implicated in the accelerated generation of epithelial cells in mice that have no B cells.

Life Sci Space Res, 1964, 2, 433 - 9
Gnotobiotic techniques and their application to spacecraft fabrication; Trexler PC; Several colonies of rats and mice have been reared totally free of viable microorganisms, some for over 10 years . A variety of other species including monkeys, dogs, sheep and swine have been maintained under the same conditions for shorter periods . This operation requires more precise control of microbic contamination than can be achieved by conventional techniques . The necessary control was realized by the use of apparatus termed "isolators" . Obtaining sterility is a biological problem but its maintenance is an engineering problem . The gnotobiotic animal has been shown to be more sensitive to some microbic contamination than in vitro culture media . Apparently complex spacecraft cannot be sterilized effectively by postassembly treatment alone . Therefore, sterile subassemblies must be made as required with final assembly in a sterile environment to avoid trapping viable organisms . This can be accomplished within isolators since there appears to be no size limitations to the apparatus developed for biological and medical applications . Repairs and testing procedures under sterile conditions are possible with the isolator system . Fabrication of spacecraft within closed sterile isolators should present no major engineering problems provided all of the components are sterilizable.

Life Sci Space Res, 1964, 2, 385 - 405
A general review of chemical sterilization in space research; Opfell JB; The unsolved problems in the biology and the chemistry of chemical sterilization are legion . This discussion of them emphasizes some interesting areas for useful research . Studies in molecular biology and cancer chemotherapy have produced information which has important implications in chemical sterilization, particularly with the alkylating and the oxidizing agents . Many of these problems must be solved soon if chemical sterilization is to play the role it should in space research by providing means for effectively controlling the rate and extent of mingling of the forms of terrestrial life with those of extraterrestrial life . In space research in the United States, recent work on chemical sterilization has laid heavy emphasis on the identification and exploration of principles of engineering sciences which control sterilization effectiveness . These principles are reviewed and their significance in chemical sterilization is discussed in terms of the engineering factors involved . Among these factors are: consequence of sterilant polymerization or reaction with the object to be sterilized; criteria for chemical sterilization process effectiveness; depletion of the concentration of the sterilant by evaporation or chemical reaction, including polymerization; obstacles to the diffusion or convection of the sterilizing chemical to the immediate vicinity of the microorganisms to be destroyed; protection of the microorganisms from direct contact with the sterilant; phase behavior of the sterilant in the environment of the sterilization process; presence of water or water vapor; the nature, sources and numbers of microorganisms which survive standard chemical sterilization processes; and the length of exposure required to sterilize at each temperature . The significance of chemical sterilization to space science has been well reviewed at the previous COSPAR symposia . Because several thorough reviews of the empirical knowledge concerning the effects of chemical agents on microorganisms have been published recently, this information is not discussed again for chemical sterilants other than the alkylating agents and several of the oxidizing agents . This lack of emphasis indicates only that these other chemicals have not been applied to a large extent in space research to date and that discussions of their sterilizing properties are readily available in other recent reviews . In the future, interest in many of these other chemical sterilants will increase as the variety of devices to be sterilized and the variety of functions which sterilized devices are expected to perform increase.

Nature, 2002 Mar 7, 416(6876), 82 - 3
Arctic microorganisms respond more to elevated UV-B radiation than CO2; Johnson D et al.; Surface ultraviolet-B radiation and atmospheric CO2 concentrations have increased as a result of ozone depletion and burning of fossil fuels . The effects are likely to be most apparent in polar regions where ozone holes have developed and ecosystems are particularly sensitive to disturbance . Polar plant communities are dependent on nutrient cycling by soil microorganisms, which represent a significant and highly labile portion of soil carbon (C) and nitrogen (N) . It was thought that the soil microbial biomass was unlikely to be affected by exposure of their associated plant communities to increased UV-B . In contrast, increasing atmospheric CO2 concentrations were thought to have a strong effect as a result of greater below-ground C allocation . In addition, there is a growing belief that ozone depletion is of only minor environmental concern because the impacts of UV-B radiation on plant communities are often very subtle . Here we show that 5 years of exposure of a subarctic heath to enhanced UV-B radiation both alone and in combination with elevated CO2 resulted in significant changes in the C:N ratio and in the bacterial community structure of the soil microbial biomass.

Med Clin (Barc), 2002 Mar 2, 118(7), 251 - 5
{Sensitization and occupational asthma in poultry workers}; Borghetti C et al.; BACKGROUND: A high prevalence of asthma has been reported in poultry farmers . Our aim was to determine air contaminants in poultry confinement buildings and the prevalence of occupational asthma in these workers . PATIENTS AND METHOD: Spanish poultry farmers who participated in the European Study Prevalence and Risk Factors for Airway Obstruction in Farmers, who worked inside confinement buildings, were enrolled in a cross-sectional study that included the measurement of air contaminants (ammonia, total dust, endotoxin and microorganisms), symptoms, skin testing with common and poultry-related allergens, and a specific bronchial challenge . The Spearman correlation coefficient was used to assess the correlation between air contaminants . RESULTS: 14/15 poultry farmers participated in the study (response index: 93.3%) . Concentrations of air contaminants in the confinement buildings {median (interquartile range)} were: 7.5 (5-20) ppm for ammonia, 2.6 (1.8-4.6) mg/m3 for total dust, 137.1 (58.6-243.9) ng/m3 for endotoxin and 7,600 (907-27,750) colony-forming units/m3 * 103 for bacteria . Endotoxin correlated with bacterial concentration (r = 0.69, p < 0.01) . 5/14 poultry farmers reported wheezing and/or asthma within the last year (35.7%), and in 2 of them spirometry demonstrated current asthma (14.3%) . Specific bronchial provocation confirmed the diagnosis of occupational asthma in one subject with current asthma sensitized to the storage mite Lepidoglyphos destructor (7.1%) . CONCLUSIONS: One third of poultry farmers working inside confinement buildings reported wheezing, that is partly attributable to occupational asthma caused by storage mites.

Res Microbiol, 2002 Jan-Feb, 153(1), 33 - 6
Molecular characterization of a Spanish isolate of Mycobacterium malmoense; Ruiz M et al.; We present the genomic characterization of a Spanish isolate of Mycobacterium malmoense . Identification and molecular characterization of the microorganism were performed using the technique of PCR-restriction fragment length polymorphism analysis (PRA) and sequencing of the 16S and the ITS spacer (16S-23S rDNA intergenic spacer region) . It was seen that our isolate is very similar to the strain described in Sweden, but differs in the sequencing of the ITS from the isolate in Great Britain . Molecular techniques facilitate a rapid diagnosis and enable a better understanding of the epidemiology of this microorganism.

OMICS, 2002, 6(1), 61 - 90
The use of accurate mass tags for high-throughput microbial proteomics; Smith RD et al.; We describe and review progress towards a global strategy that aims to extend the sensitivity, dynamic range, comprehensiveness, and throughput of proteomic measurements for microbial systems based upon the use of polypeptide accurate mass tags (AMTs) produced by global protein enzymatic digestions . The two-stage strategy exploits high accuracy mass measurements using Fourier transform ion cyclotron resonance mass spectrometry (FTICR) to validate polypeptide AMTs for a specific organism, from potential mass tags tentatively identified using tandem mass spectrometry (MS/MS), providing the basis for subsequent measurements without the need for routine MS/MS . A high-resolution capillary liquid chromatography separation combined with high sensitivity, and high-resolution accurate FTICR measurements is shown to be capable of characterizing polypeptide mixtures of more than 10(5) components, sufficient for broad protein identification using AMTs . Advantages of the approach include the high confidence of protein identification, its broad proteome coverage, and the capability for stable-isotope labeling methods for precise relative protein abundance measurements . The strategy has been initially evaluated using the microorganisms Saccharomyces cerevisiae and Deinococcus radiodurans . Additional developments, including the use of multiplexed-MS/MS capabilities and methods for dynamic range expansion of proteome measurements that promise to further extend the quality of proteomics measurements, are also described.

Life Sci Space Res, 1964, 2, 305 - 10
Effect of ultraviolet radiation on microorganisms as a principal extremal factor of space environment; Fedorova RI; 1 . The problem of the transport of viable germs through interplanetary space has recently acquired special interest arising from the necessity of sterilization of space ships . 2 . Of all cosmic extremal factors ultraviolet solar radiation is the most dangerous for microorganisms . The intensity of the most bactericidal region of ultraviolet radiation at a distance of one astronomical unit from the sun is 2 x 10(3) erg per cm2 . 3 . Sensitivity of microorganisms to ultraviolet rays varies in a large range . For the most resistant forms the lethal dose is approximately 440000 erg per cm2 . 4 . The penetrating capacity of ultraviolet radiation is very low, and therefore, even insignificant amounts of organic or mineral substances may protect the bacterial cell . 5 . Dust particles of terrestrial origin carrying on themselves bacterial spores exert upon the latter a protective effect against ultraviolet radiation.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Nov-Dec, (6), 99 - 104
{Culture media as artificial environment ensuring the growth and development of microorganisms}; Akhapkina IG et al.; Literature review is presented on culture media that are regarded as artifical environment for microorganisms permitting modeling of various conditions for revealing cultural, morphological and biochemical properties, characteristic for a given microorganism, as well as manipulations with these conditions.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Nov-Dec, (6), 20 - 4
{Stimulating and inhibiting effect of Spirulina platensis on microorganisms}; Gorobets OB et al.; The blue-green microalga (cyanobacterium) S . platensis and the complex of its metabolites in the culture fluid being added into nutrient agar in doses of 0.01, 0.1 and 10 mg/ml, may produce a stimulating and inhibiting effect on microorganisms . In Spirulina biomass and in culture fluid substances which may be associated with the stimulation or inhibition of the growth of some microorganisms are supposedly contained . The manifestation of stimulating or inhibiting properties depended on the concentration of S . platensis and the complex of its metabolites, as well as on sterilization methods of nutrient media supplemented with these substances.

J Agric Food Chem, 2002 Mar 13, 50(6), 1625 - 30
Release of ferulic acid from oat hulls by Aspergillus ferulic acid esterase and trichoderma xylanase; Yu P et al.; Oat hulls, an agricultural byproduct, contain a relatively high amount of ferulic acid (FA; 4-hydroxy-3-methoxycinnamic acid), which is believed to be inhibitory to oat hull biodegradability by rumen microorganisms . In this paper, Aspergillus ferulic acid esterase (FAE) was investigated for its ability to release FA from oat hulls . The objectives were to determine the effects of particle size of oat hulls (ground to pass through 1 mm and 250 microm screens and a 100 microm sieve) on release of FA by FAE both in the presence and in the absence of Trichoderma xylanase . The results show that the release of FA by FAE was dependent upon the particle size of oat hulls (< or = 250 microm) . In the absence of Trichoderma xylanase, little FA was released by FAE . In the presence of Trichoderma xylanase, there was a significant release of FA by FAE, indicating a synergistic interaction between FAE and Trichoderma xylanase on release of FA from oat hulls . These results indicate that FAE is able to break the ester linkage between FA and the attached sugar, releasing FA from oat hulls . This may leave the remainder of the polysaccharides open for further hydrolytic attack by rumen microorganisms . It is likely that removing FA from oat hulls could improve rumen biodegradability, thus improving the nutritional value of oat hulls.

J Agric Food Chem, 2002 Mar 13, 50(6), 1435 - 40
Microbial inactivation of paprika by a high-temperature short-X time treatment . Influence on color properties; Almela L et al.; High-temperature short-time (HTST) treatments have been used to destroy the bioburden of paprika . With this in mind, we have designed a device to treat samples of paprika with a gas whose temperature, pressure, and composition can be selected . Temperatures and treatment times ranged from 130 to 170 degrees C and 4 to 6 s, respectively . The survival of the most commonly found microorganisms in paprika and any alteration in extractable and superficial color were examined . Data showed that the optimum HTST conditions were 145 degrees C, 1.5 kg/cm2 of overpressure, 6 s operation time, and a thermal fluid of saturated steam . No microbial growth was detected during storage after thermal treatment . To minimize the color losses, treated (HTST) paprika samples should be kept under refrigeration.

Tree Physiol, 2002 Mar, 22(4), 231 - 8
Characterization of a Pinus pinaster cDNA encoding an auxin up-regulated putative peroxidase in roots; Charvet-Candela V et al.; As part of a study to identify host plant genes regulated by fungal auxin during ectomycorrhiza formation, we differentially screened a cDNA library constructed from roots of auxin-treated Pinus pinaster (Ait.) Sol . seedlings . We identified three cDNAs up-regulated by auxin . Sequence analysis of one of these cDNAs, PpPrx75, revealed the presence of an open reading frame of 216 amino acids with the characteristic consensus sequences of plant peroxidases . The deduced amino acid sequence showed homology with Arabidopsis thaliana (L.) Heynh., Arachis hypogaea L . and Stylosanthes humilis HBK cationic peroxidases . Amino acid sequence identities in the conserved domains of plant peroxidases ranged from 60 to 100% . In PpPrx75, there are five cysteine residues and one histidine residue that are found at conserved positions among other peroxidases . A potential glycosylation site (NTS) is present in the deduced sequence . Phylogenetic analysis showed that PpPrx75 is closely related to two A . thaliana peroxidases . The PpPrx75 cDNA was induced by active auxins, ethylene, abscisic acid and quercetin, a flavonoid possibly involved in plant-microorganism interactions . Transcript accumulation was detected within 3 h following root induction by auxin, and the amount of mRNA increased over the following 24 h . The protein synthesis inhibitor cycloheximide did not inhibit indole-3-acetic acid-induced transcript accumulation, suggesting that PpPrx75 induction is a primary (direct) response to auxin . This cDNA can be used to study expression of an auxin-regulated peroxidase during ectomycorrhiza formation.

Appl Environ Microbiol, 2002 Mar, 68(3), 1157 - 64
Interactions among plant species and microorganisms in salt marsh sediments; Burke DJ et al.; The interactions among Spartina patens and sediment microbial populations and the interactions among Phragmites australis and sediment microbial populations were studied at monotypic sites in Piermont Marsh, a salt marsh of the Hudson River north of New York, N.Y., at key times during the growing season . Arbuscular mycorrhizal fungi (AMF) effectively colonized S . patens but not P . australis, and there were seasonal increases and decreases that coincided with plant growth and senescence (17 and 6% of the S . patens root length were colonized, respectively) . In sediment samples from the Spartina site, the microbial community and specific bacterial populations were at least twice as large in terms of number and biomass as the microbial community and specific bacterial populations in sediment samples from the Phragmites site, and peak values occurred during reproduction . Members of the domain Bacteria, especially members of the alpha-, gamma-, and delta-subdivisions of the Proteobacteria, were the most abundant organisms at both sites throughout the growing season . The populations were generally more dynamic in samples from the Spartina site than in samples from the Phragmites site . No differences between the two sites and no differences during the growing season were observed when restriction fragment length polymorphism analyses of nifH amplicons were performed in an attempt to detect shifts in the diversity of nitrogen-fixing bacteria . Differences were observed only in the patterns generated by PCR or reverse transcription-PCR for samples from the Spartina site, suggesting that there were differences in the overall and active populations of nitrogen-fixing bacteria . Regression analyses indicated that there was a positive interaction between members of the delta-subdivision of the Proteobacteria and root biomass but not between members of the delta-subdivision of the Proteobacteria and macroorganic matter at both sites . In samples from the Spartina site, there were indications that there were bacterium-fungus interactions since populations of members of the alpha-subdivision of the Proteobacteria were negatively associated with AMF colonization and populations of members of the gamma-subdivision of the Proteobacteria were positively associated with AMF colonization.

BMC Oral Health . 2002;2(1):1.
A new chemical formulation for control of dental unit water line contamination: An 'in vitro' and clinical 'study'
Montebugnoli L, Dolci G.
BACKGROUND: Water delivered by dental units during routine dental practice is highly contaminated . The aim of this study is to evaluate the efficacy of a new chemical solution flushed through Dental Unit Water Lines (DUWL) for the control of contamination inside dental units . MATERIALS AND METHODS: Six old dental units equipped with a device designed to automatically flush disinfecting solutions through the water system (Castellini Autosteril) were selected . Water samples from DUWL effluents were collected in each dental unit for 10 randomly selected days, before and after a 5 minute DUWL disinfecting cycle with TetraAcetylEthileneDiamine (TAED) and persalt (Ster4spray produced by Farmec spa, and distributed by Castellini spa) . Water samples were plated in R2A Agar and cultured at room temperature for 7 days, and the total number of heterotrophic microorganisms counted and expressed in Log10 CFU/mL A general linear model was fitted and multiple regression ANOVA for repeated measures was used for the statistical analysis . RESULTS: The mean contamination in DUWL effluent at baseline was 5.45 &#PlusMinus; 0.35 CFU/mL (range 4.79 to 5.93 CFU/mL) . When water samples were tested "in vitro" against the chemical, no growth of heterotrophic bacteria was detected after a 5 minute contact in any of the water samples tested . After undergoing a 5 minute disinfecting cycle with the chemical, DUWL mean contamination in water effluents was 2.01 &#PlusMinus; 0.32 CFU/mL (range 1.30 to 2.74 CFU/mL) (significant difference with respect to baseline) . CONCLUSIONS: An inbetween patient disinfecting procedure consisting of flushing DUWL with TAED and persalt equivalent to 0.26% peracetic acid could be useful in routine dental practice for cross-contamination control.

BJU Int, 2002 Mar, 89(4), 426 - 32
Knowledge and attitudes of Korean parents towards their son's circumcision: a nationwide questionnaire study; Oh SJ et al.; OBJECTIVE: To evaluate knowledge about the foreskin and circumcision, and to understand the attitudes of parents to circumcision in Korea, where circumcision in childhood is widely practised with no particular religious or medical background . SUBJECTS AND METHODS: A nationwide study involving questionnaires was conducted on 5500 parents with at least one son attending elementary school . Responses were obtained from one of the parents . RESULTS: The response rate was 76.1% (4183); circumcision was most common in boys when aged 11 years, followed by neonatal circumcision . Of the parents, 91.3% believed that circumcision is necessary, while 2.1% believed it to be unnecessary . The principal reasons given for circumcision were 'to improve penile hygiene' (82.4%), followed by 'to improve future sexual potency' (7.5%) . Among those who did not believe circumcision to be necessary, the most common reason was the expectation of spontaneous retraction of the prepuce with age (55.1%) . Most (88.4%) of the parents believed that smegma is not a clean material, and is infected by microorganisms . Most parents (80.6%) thought that circumcision would prevent genital tract infection of the future spouse . Peer pressure was one of the most influential factors in deciding upon circumcision; 41.9% of the parents were anxious that their child might be ridiculed by his peer group unless he was circumcised, while 27.4% of the parents believed that their child might be ridiculed if he was circumcised . Mothers were more positive about circumcision than fathers (P < 0.05) . Parents with a higher education and higher socio-economic status were also more positive about circumcision (P < 0.05) . Mothers were prone to emphasize improved sexual potency (P < 0.05) . There was no significant difference in response between urban and rural areas . CONCLUSION: This study indicates that common beliefs held by parents about the prepuce or circumcision differ significantly from current medical knowledge, and these beliefs have a major influence on the practice of circumcision in Korea . More clinical research on the natural history of the foreskin is needed, and it is critical that both children and parents are informed about the potential benefits and disadvantages of circumcision.

J Environ Monit, 2002 Feb, 4(1), 166 - 74
Personal exposures and microenvironmental concentrations of particles and bioaerosols; Toivola M et al.; The aim of this study was to compare the personal exposure to particles and bioaerosols with that measured by stationary samplers in the main microenvironments, i.e., the home and the workplace . A random sample of 81 elementary school teachers was selected from the 823 teachers working for two councils in eastern Finland for the winter time measurement period . Bioaerosol and other particles were collected on filters by button samplers using personal sampling and microenvironmental measurements in homes and workplaces . The 24-hour sampling period was repeated twice for each teacher . Particle mass, absorption coefficient of the filter and the concentration of viable and total microorganisms were analyzed from each filter . In this paper, the study design, quality assurance principles and results of particle and bioaerosol exposure are described . The results show that particle mass concentrations, absorption coefficient and fungi were higher in personal exposure samples than in home and workplace samples . Furthermore, these concentrations were usually lower in the home than in the workplace . Bacterial concentrations were highest in heavily populated workplaces, while the viable fungi concentrations were lowest in workplaces . The fungi and bacteria results showed high variation, which emphasises the importance of quality assurance (duplicates and field blanks) in the microbial field measurements . Our results indicate that personal exposure measurements of bioaerosols in indoor environments are feasible and supplement the information obtained by stationary samplers.

Zh Obshch Biol, 2001 Nov-Dec, 62(6), 472 - 95
{Genetic-evolutionary basis of symbiosis doctrine}; Provorov NA; The author presents the current notion of symbiosis as one of the main adaptation of an organism to changeable environment . Symbiosis is considered as a super organism genetic system within which there are different interactions (including mutualism and antagonism) . Genetic integration of symbiotic partners can be realized as cross regulation of their genes, exchange of gene products (proteins, RNA), gene amplification and sometimes gene transfer between organisms . On the phenotypic level these processes result in signal interactions, integration of partner metabolic systems and development of symbiotic organs . Co-evolution is considered as an assemblage of micro- and macroevolution processes basing on pre-adaptations and proceeding under influence of different forms of natural selection (individual, frequency-depended and kin selection) . Symbiosis can be compared with sexual process since both are the forms of organism integration characterized by different genetic mechanisms and evolutionary consequences . The genome evolution in symbiotic microorganisms can proceed by: 1) simplification of genome in obligate symbiosis (loss of genes that are necessary for independent existence, transfer of some genes to the host organism); 2) complication of genome in facultative symbiosis (increase in genome plasticity, structural and functional differentiation of genome into systems controlling free-living and symbiotic parts of life cycle) . Most of symbiotic interactions are correlated to an increase in genetic plasticity of an organism that can lead to evolutionary saltations and origin of new forms of life.

Cochrane Database Syst Rev . 2002;(1):CD002929.
Disposable surgical face masks for preventing surgical wound infection in clean surgery; Lipp A et al.; BACKGROUND: Surgical face masks were originally developed to contain and filter droplets of microorganisms expelled from the mouth and nasopharynx of healthcare workers during surgery, thereby providing protection for the patient . However there are several ways in which surgical face masks could potentially contribute to contamination of the surgical wound . OBJECTIVES: To identify and review all randomised controlled trials evaluating disposable surgical face masks worn by the surgical team during clean surgery to prevent post-operative surgical wound infection . SEARCH STRATEGY: All relevant publications about disposable surgical face masks were sought through the Specialised Trials Register of the Cochrane Wounds Group (March 2001) . Manufacturers and distributors of disposable surgical masks as well as professional organisations including the National Association of Theatre Nurses and the American Operating Room Nurses Association were contacted for details of unpublished and ongoing studies . SELECTION CRITERIA: Randomised controlled trials (RCTs) and quasi-randomised controlled trials comparing the use of disposable surgical masks with the use of no mask were included . DATA COLLECTION AND ANALYSIS: Data were extracted independently by AL and PE . MAIN RESULTS: Two randomised controlled trials were included involving a total of 1453 patients . In a small trial there was a trend towards masks being associated with fewer infections, whereas in a large trial there was no difference in infection rates between the masked and unmasked group . Neither trial accounted for cluster randomisation in the analysis . REVIEWER'S CONCLUSIONS: From the limited results it is unclear whether wearing surgical face masks results in any harm or benefit to the patient undergoing clean surgery.

Clin Microbiol Infect, 1998 Jan, 4 Suppl 4, S1 - S6
Epidemiology of Chlamydia pneumoniae; Blasi F et al.; Chlamydia pneumoniae is the most commonly occurring intracellular bacterial pathogen . It is frequently involved in respiratory tract infections and to a lesser degree in extrapulmonary diseases . According to seroepidemiologic surveys, C . pneumoniae infection seems to be both endemic and epidemic . Such studies indicate that C . pneumoniae infection is widespread, with frequent reinfection during a lifetime . In Western countries the highest rate of new infections occurs between the ages of 5 and 15 . The antibody prevalence worldwide is higher in adult males than in females . Currently available data suggest that C . pneumoniae is primarily transmitted from human to human without any animal reservoir . Transmission seems to be inefficient, although household outbreaks with high transmission rates are reported . Most reports rank C . pneumoniae among the three most common etiologic agents of community-acquired pneumonia, with an incidence ranging from 6% to 25%, and generally presenting a mild and, in some cases, self-limiting clinical course . Recent reports also indicate a possible role for C . pneumoniae in severe forms of community-acquired pneumonia and in respiratory infections in immunocompromised patients . C . pneumoniae infection has also been implicated in the pathogenesis of asthma in both adults and children . The hypothesis that C . pneumoniae infection could lead to asthma is based on clinical studies and on the evidence of specific IgE production, direct epithelial damage, induction of T-cell immunopathologic diseases, and vascular smooth cell infection . Chronic C . pneumoniae infection seems to be common in patients with chronic bronchitis whether exacerbated or not, and is characterized by a strong humoral immune response to this intracellular microorganism, which is present in the majority of patients with severe chronic bronchitis . More than 60% of subjects with chronic bronchitis have specific C . pneumoniae antibody titers, and the microorganism may be identified by culture or PCR in almost 40% of these patients . This pathogen has also been recently associated with atherosclerosis and coronary heart disease (CHD) . Seroepidemiological evidence indicates that the majority of patients with CHD present an anti-C . pneumoniae antibody pattern consistent with chronic infection . Furthermore, C . pneumoniae has been detected in atherosclerotic coronary plaques by several methods, including immunocytochemistry, transmission electron microscopy and molecular biology techniques . Recently, we detected C . pneumoniae DNA in a high percentage (51%) of aortic aneurysm plaques . Moreover, our serologic data support the hypothesis that a chronic C . pneumoniae antibody pattern may be a possible risk marker for atherosclerosis . Recently, C . pneumoniae has been isolated by culture from the coronary artery of a patient with coronary atherosclerosis, providing direct evidence of the presence of viable organisms in atheromatous lesions . Moreover, we recently demonstrated an association between C . pneumoniae reinfection and acute myocardial infarction.

EDTNA ERCA J, 2001 Jul-Sep, 27(3), 140 - 2, 146
Containers for waste causing peritonitis in CAPD patients; Alscher DM et al.; Fungal infection is a rare cause of peritonitis in patients on chronic peritoneal dialysis . In this population, fungi are found in less than 2 percent of all cases of primary episodes of peritonitis (1) . More often, a primary bacterial peritonitis treated with antibiotic therapy leads to secondary fungal infection (2) . Candida species cause 74.5% of the episodes of fungal peritonitis (2) . The fungi invade the peritoneal cavity from the skin peri- or intraluminally through the catheter (3) . Filamentous fungi are rare (4,5).Treatment of fungal peritonitis commonly consists of removal of catheter and antifungal drugs (3) . Here we describe two cases of fungal peritonitis caused by mycelial fungi, where the source of the microorganism could be special containers used for biological waste, which are popular in Germany

J Steroid Biochem Mol Biol, 2002 Jan, 80(1), 131 - 3
Conversion of liposomal 4-androsten-3,17-dione by A . simplex immobilized cells in calcium pectate; Llanes N et al.; Arthrobacter simplex ATCC 6946 free and immobilized cells were assayed for their ability to convert 4-androsten-3,17-dione (AD) to 1,4-androstadien-3,17-dione (ADD) in aqueous and liposomal media . Bioconversions were carried out in a 100 ml flask containing 25 ml of AD liposomal or aqueous medium for 3h, and AD concentrations ranging from 0.3 to 1.0 mM were tested . AD/ADD ratios in samples were determined by HPLC . Biotransformation of substrate entrapped in multilamellar vesicles (MLV) was demonstrated to be better than the corresponding free form . In the former case, 2h were necessary to completely bioconvert 1 mM AD . By contrast, 3h were needed to reach 50% bioconversion in (4%) ethanol medium containing 0.63 mM AD . The liposomal medium allows us to perform steroid conversions at high concentrations of AD, reusing immobilized cells in suitable conditions which are non-toxic for microorganisms.

Phytochemistry, 2002 Mar, 59(6), 579 - 96
The destruxins: synthesis, biosynthesis, biotransformation, and biological activity; Pedras MS et al.; Destruxins, secondary metabolites first reported in 1961, are cyclic hexadepsipeptides composed of an alpha-hydroxy acid and five amino acid residues . The name "destruxin" is derived from "destructor" from the species Oospora destructor, the entomopathogenic fungus from which these metabolites were first isolated . Individual destruxins differ on the hydroxy acid, N-methylation, and R group of the amino acid residues; where established, the configurations of the amino acid residues are S, and those of the hydroxy acids are R . Destruxins exhibit a wide variety of biological activities, but are best known for their insecticidal and phytotoxic activities . The great interest in destruxins derives from their potential role as virulence factors in fungi, whether such microorganisms are useful insect biocontrol agents or detrimental, causing great plant disease epidemics . Reports on isolation, chemical structure determination, total synthesis, transformation by diverse organisms, and biological activity of destruxins and related metabolites are reviewed for the first time.

Clin Microbiol Infect, 1996 Mar, 1 Suppl 1, S1 - S2
Definition of intracellular pathogens; Orfila J; Most microorganisms are destroyed by the host tissues through processes which usually involve phagocytosis and lysosomal disruption . However, some organisms are capable of growing inside macrophages and avoiding destruction . This paper describes the normal phagocytic process and how it is avoided by certain viruses, bacteria and protozoa, and the complex cycle of intracellular replication by species of Chlamydia.

Clin Microbiol Infect, 1998 Feb, 4(2), 88 - 93
Evaluation of the BacT/Alert and VITAL blood culture systems for the diagnosis of bacteremia; Fontanals D et al.; OBJECTIVE: To evaluate the detection of bacterial growth in the BacT/Alert (Organon Teknika) and VITAL (bioMerieux) automated blood culture systems . METHODS: In accordance with the protocol of study, 1021 blood sample pairs for culture were obtained from adult patients admitted to the Emergency Room and Intensive Care Unit . RESULTS: In total, 139 (13.6%) clinically significant blood cultures were detected, of which 79 (56.8%) were detected by both systems, 48 (34.5%) only by BacT/Alert and 12 (8.6%) only by VITAL (P cent0.0001) . The BacT/Alert system detected positive blood cultures more rapidly for all groups of microorganisms . The VITAL system showed six false-negative blood cultures, while the BacT/Alert system showed none (P50.03) . There was no significant difference between the number of false-positive blood cultures detected by the two systems . CONCLUSIONS: In our study, overall the BacT/Alert system achieved a better recovery of microorganisms than the VITAL system.

Clin Microbiol Infect, 1997 Jun, 3(3), 345 - 351
Comparison of BacT/Alert and BACTEC NR 860 blood culture systems in a laboratory not continuously staffed; Riest G et al.; OBJECTIVE: To evaluate the performance of continuously working blood culture systems in a discontinuous laboratory system . METHODS: The systems used were BacT/Alert (Organon Teknika Corp., Durham, NC) and BACTEC NR 860 (Becton Dickinson Diagnostic Instruments, Sparks, Md) in a comparison in a laboratory staffed 8 1/2 h on Mondays to Fridays and 4 1/2 h on Saturdays . Blood culture bottles (BacT/Alert aerobic and anaerobic, BACTEC NR 26 A and NR 27 A) were received thrice daily . RESULTS: From 1824 pairs of blood culture vials, 110 clinically significant microorganisms were recovered by both BACTEC and BacT/Alert, 43 by BACTEC alone, and 33 by BacT/Alert alone . The differences between the systems in total recovery and in recovery of individual species were not statistically significant . The average detection times were 13.36 h for BACTEC and 13.93 h for BacT/Alert (P>0.1) . These times represent only 35.6% (BACTEC) and 32.6% (BacT/Alert) of the total timespans from collection of blood to informing the ward of a positive result (tcrd, clinically relevant detection time) . If 24 h per day blood culture processing conditions and continuous transport of vials to the laboratory had been available, these percentages would have risen to 87% (BACTEC) and 87.5% (BacT/Alert) . Under such 'ideal' conditions, ttrd could have been reduced by 22.16 h using BACTEC and by 26.81 h using BacT/Alert . The BacT/Alert system showed more false-positive results than the BACTEC system (80 (4.39%) versus 23 (1.26%), P<0.001) . CONCLUSIONS: No time benefit for detection of positive blood cultures is gained with continuously measuring systems, if loading and processing of vials is organized discontinuously, as in our laboratory.

Arch Intern Med, 2002 Feb 25, 162(4), 477 - 9
Identification of Cardiobacterium hominis by broad-range bacterial polymerase chain reaction analysis in a case of culture-negative endocarditis; Nikkari S et al.; Culture-negative bacterial endocarditis may be attributed to fastidious microorganisms, prior institution of antibiotic treatment, or both . We describe a case of culture-negative endocarditis in which a modified Steiner stain revealed bacterial structures in the resected heart valve material . Prompted by this finding, broad-range polymerase chain reaction (PCR) amplification of small-subunit ribosomal DNA (16S rDNA) was performed, and Cardiobacterium hominis sequences were detected . This case demonstrates the usefulness of both the Steiner stain and broad-range direct molecular amplification as supplemental diagnostic tools in identification of otherwise unexplained infections.

C R Biol, 2002 Jan, 325(1), 75 - 6
Inactivation of the BSE agent; Taylor D; In the studies carried out so far, the BSE agent has proved to be just as resistant as other TSE agents to inactivation by procedures such as autoclaving or exposure to sodium hydroxide that are effective with conventional microorganisms . However, in common with other TSE agents, the BSE agent appears to be effectively inactivated by exposure to sodium hypochlorite solutions containing high levels of available chlorine . Not surprisingly, the BSE agent has been found to survive at least some of the rendering processes that were used to process tissues discarded by abattoirs in the EU during the early 1980s . Despite the survival of BSE infectivity after autoclaving or exposure to sodium hydroxide, it is known that combining these procedures results in a very reliable degree of inactivation for TSE agents generally . The combination of heat and alkali has also been shown to be effective with a mouse-passaged strain of BSE agent, even at a temperature of only 100 degrees C for a minute . Also, in carrying out BSE-spiked validation studies relating to the safety of bone-derived gelatin, it has also been found that the exposure of acid-treated bone (which is free from any obvious remains of fatty or proteinaceous tissue) to 0.3 M sodium hydroxide for two hours knocks out any residual BSE infectivity.

Int J Gynecol Cancer, 2002 Jan-Feb, 12(1), 3 - 17
Immunology of the peritoneal cavity: relevance for host-tumor relation; Melichar B et al.; The peritoneal membrane, formed by a single layer of mesothelial cells, lines the largest cavity of the human body . Anatomic structures of the peritoneal cavity, along with resident leukocyte populations, play an important role in the defense against microorganisms invading by breaching the gut integrity or ascending through the female genital tract . Local immune mechanisms in the peritoneal cavity are also important in patients undergoing peritoneal dialysis and in women with endometriosis . There is now extensive evidence demonstrating the significance of peritoneal immune mechanisms in the control of metastatic spread . Leukocytes belonging to both the innate and adaptive immune systems are present in the peritoneal cavity of normal subjects as well as in patients with intra-abdominal cancer . There is now increased understanding of the mechanisms that not only allow the tumor cells to escape the detection and destruction by the host immune system, but also to use the inflammatory mechanisms to promote tumor growth and spread inside the peritoneal cavity . Malignant ascites represents a model for the study of the interaction between tumor cells and the host immune system as well for the analysis of the tumor microenviroment . The peritoneal immune system may be stimulated by intraperitoneal administration of biologic agents . This peritoneal immunotherapy may be used for palliation of malignant ascites, or as a consolidation strategy in patients with minimal residual disease.

Amino Acids, 2001 Dec, 21(4), 383 - 91
Catabolism of methionine and threonine in vitro by mixed ruminal bacteria and protozoa; Or-Rashid MM et al.; In vitro studies were conducted to examine the metabolism of methionine (Met) and threonine (Thr) using mixed ruminal bacteria (B), mixed ruminal protozoa (P), and a combination of these two (BP) . Rumen microorganisms were collected from fistulated goats fed with lucerne cubes (Medicago sativa) and a concentrate mixture twice a day . Microbial suspensions were anaerobically incubated with or without 1 mM each of the substrates at 39 degrees C for 12h . Met, Thr and their related amino compounds in both the supernatants and microbial hydrolyzates of the incubation were analyzed by HPLC . Met was degraded by 58.7, 22.1, and 67.3% as a whole in B, P, and BP suspensions, respectively, during 12h incubation . In the case of Thr, these values were 67.3, 33.4, and 76.2% in B, P, and BP, respectively . Met was catabolized by all of the three microbial suspensions to methionine sulfoxide and 2-aminobutyric acid . Catabolism of Thr by B and BP resulted in the production of glycine and 2-aminobutyric acid, while P produced only 2-aminobutyric acid . From these results, the existence of diverse catabolic routes of Met and Thr in rumen microorganisms was indicated.

J Antibiot (Tokyo), 2001 Dec, 54(12), 1066 - 71
Biosynthesis of the thiazolylpeptide antibiotic GE2270; De Pietro MT et al.; The biosynthesis of the antibiotic GE2270 in the producing microorganism Planobispora rosea was investigated by adding labelled amino acid precursors . Efficient incorporation of glycine and serine was observed, leading to specific enrichments of selected positions of the thiazole, oxazoline and pyridine rings . Furthermore, efficient enrichment of the C-, N- and O-methyl groups was detected . These results indicate that GE2270 is made through a biosynthetic route similar to that determined for other thiazolylpeptides . At the same time, the result point to an efficient route for the conversion of glycine into serine and methyl equivalents in Planobispora rosea.

Mol Cell Biochem, 2001 Dec, 228(1-2), 1 - 7
The use of calcium blockers to study biochemical behaviour of Saccharomyces cerevisiae cells; de Souza Pereira R; Calcium channel blockers, currently used in clinical medicine, from the dihydropyridine class (nifedipine and nimodipine) were used to study biochemical behaviour of Saccharomyces cerevisiae cells . At a concentration of 200 microM, both drugs partially blocked calcium uptake from an extracellular medium . On the other hand, nifedipine (300 microM) induced a collapse of the membrane potential (delta psi) of these cells, while nimodipine, at the same concentration, produced a similar but less intense effect . Probably, the small difference in the chemical structure between these drugs may explain the same effect with different intensity . With these experiments was possible to observe that these microorganisms have type 'L' calcium channels.

Huan Jing Ke Xue, 2001 Nov, 22(6), 91 - 4
{Effect of toluene on urease activity of Lou soil}; He W et al.; Urease activity affected by toluene studying results showed as follows: Toluene had little action on Jackbean urease activity, but can remarkably increase soil urease activity in lower toluene concentrations and shorter time, the increasing variation was 1.48-3.96 times than without toluene treatment in seven soil samples tested . The variation was less than 3.98% when sterilized soil samples adsorbed the Jackbean urease . It may be because toluene can kill soil microorganism, release urease within cell and subsequently urease fixed into soil organic-inorganic colloidal . Different soil pH and microbiology can lead to vary differently for urease activity.

Infect Immun, 2002 Mar, 70(3), 1481 - 7
Expression of surfactant protein D in the human gastric mucosa and during Helicobacter pylori infection; Murray E et al.; Helicobacter pylori establishes persistent infection of gastric mucosa with diverse clinical outcomes . The innate immune molecule surfactant protein D (SP-D) binds selectively to microorganisms, inducing aggregation and phagocytosis . In this study, we demonstrated the expression of SP-D in gastric mucosa by reverse transcription-PCR and immunohistochemical analysis . SP-D is present at the luminal surface and within the gastric pits, with maximal expression at the surface . Levels of expression are significantly increased in H . pylori-associated gastritis compared to those in the normal mucosa . Immunofluorescence microscopy was used to demonstrate binding and agglutination of H . pylori by SP-D in a lectin-specific manner . These activities resulted in a 50% reduction in the motility of H . pylori, as judged on the basis of curvilinear velocity measured by using a Hobson BacTracker . Lipopolysaccharides extracted from three H . pylori strains were shown to bind SP-D in a concentration-dependent manner, and there was marked variation in the avidity of binding among the strains . SP-D may therefore play a significant role in the innate immune response to H . pylori infection.

J Vet Med Sci, 2002 Jan, 64(1), 57 - 62
Postweaning multisystemic wasting syndrome of pigs in Korea: prevalence, microscopic lesions and coexisting microorganisms; Kim J et al.; A retrospective study was performed on natural cases of postweaning multisystemic wasting syndrome (PMWS), recorded from January 1999 to December 2000, to determine the prevalence, microscopic lesions, and other coexisting pathogens associated with PMWS . PMWS is diagnosed based on three criteria: the presence of clinical signs (retardation of growth), characteristic microscopic lesions (granulomatous inflammation and inclusion body), and the presence of porcine circovirus (PCV)-2 within these lesions . One hundred and thirty three (8.1%) of the 1634 pigs submitted from 1243 pig farms were diagnosed for PMWS . The affected pigs were from 25 to 120 days old, the majority (78 cases, 58.6%) being 60 to 80 days old . PMWS occurred each month during the two-year study period, but the incidence peaked in May (38 cases, 28.6%), followed by April (18 cases, 13.5%) and June (13 cases, 9.8%) . The most consistent and characteristic lesions were multifocal, granulomatous inflammation in lymph nodes, liver and spleen, characterized by infiltration of epithelioid macrophages and multinucleated giant cells . The majority of cases (113 cases, 85.0%) was dual infection with other pathogens . The combination of PCV-2 and Hemophilus parasuis (43 cases, 32.3%) was shown to be the most prevalent followed by PCV-2 and porcine reproductive and respiratory syndrome virus (39 cases, 29.3%) . The consistent presence of PCV-2, but lower prevalence of other viral and bacterial pathogens in all pigs examined with PMWS, has led to the speculation that PCV-2 is the etiological agent causing PMWS.

Berl Munch Tierarztl Wochenschr, 2002 Jan-Feb, 115(1-2), 16 - 23
{The effectiveness of rare earth elements as a possible alternative growth promoter for pigs and poultry}; Schuller S et al.; Mixtures of Rare Earth Elements (REE) have been used as animal growth-promoters on a large scale in China during the last 20 years . Numerous studies carried out in China claim it produces quite sensational growth-promoting effects in all categories of farm animals . To explore the question of whether REE's might prove suitable as a growth-promoter under western keeping conditions, feeding experiments were performed on pigs and poultry . The animals received a typical diet, supplemented with REE salts in concentrations between 75 and 300 mg/kg feed . Weight-gain, feed-intake, feed-conversion and (where applicable) laying parameters were observed . It was shown that in pigs receiving feed supplemented with REEs, an increase in daily weight gain of up to 19% and an improvement in feed-conversion of up to 11% can be achieved, whereas, for poultry, no positive effects on growth or productivity of the animals could be observed . Testing of important organs via Neutron Activating Analysis (NAA) showed a minute accumulation of REE, principally in liver and bones . Analysis of the poultry gut-flora, using selective media, showed that the main microorganism populations of the alimentary canal were unaffected by feed-supplementation with REE.

Prikl Biokhim Mikrobiol, 2002 Jan-Feb, 38(1), 5 - 13
{The antiviral activity of chitosan (review)}; Chirkov SN; Data on the inhibitory effect of chitosan on viral infections in animals, plants, and microorganisms are reviewed . The effects of the physicochemical parameters and structure of chitosan on its antiviral activity are analyzed . Possible mechanisms of the inhibitory effect of chitosan on viral infections are discussed.

J Adv Nurs, 2002 Feb, 37(3), 227 - 33
Efficacy of the Shinki bioclean room for preventing infection in neutropenic patients; Shinjo K et al.; AIM OF THE STUDY: To investigate the effectiveness of a new type of bioclean room named Shinki bioclean room (SBCR) for the prevention of infection during neutropenia after intensive chemotherapy in comparison with a standard laminar air flow room (LAFR) . BACKGROUND: Recently, a new industrial technology, wherein a dust-free and aseptic environment is created by circulating air containing nanometre order ultra fine water droplets with abundant negative air ions, has been developed in Japan . METHODS: The air cleanliness of SBCR was examined by measuring airborne particles and microorganisms . Bacteriological samples for environment culture were taken by means of exposed settle-plates . In addition, the frequency of pneumonia and fever higher than 38 degrees C were examined in 34 patients with acute leukaemia who received intensive chemotherapy in SBCR or LAFR . RESULTS: The number of airborne particles (> or = 0.5 microm) was 70 particles/ft3, and that of airborne microorganisms was 0.0 colony forming unit/ft3 in SBCR, and neither bacteria nor fungi were detected . The numbers of colonies of bacteria and fungi on air settle-plates were fewer in the SBCR than in the LAFR regardless of the presence of patients or the nurse entering . The frequency of pneumonia during chemotherapy for acute leukaemia was lower in the SBCR group (0%, 0/19 cases) than in the LAFR group (27%, 4/15 cases) (P=0.0294) and the frequency of fever higher than 38 degrees C also tended to be lower in the SBCR group (53%, 10/19 cases) than in the LAFR group (80%, 12/15 cases) (P=0.0973) . CONCLUSION: The SBCR is equal or superior to LAFR in preventing infection during neutropenia . Other advantages for SBCR are a low level of noise (40 dB), easy control of temperature and humidity, and efficient removal of odour . In addition to the quiet and comfortable atmosphere, expected favourable effects of negative air ions may give higher quality of life for patients in SBCR than those in LAFR . Further studies will be needed to examine the safety, benefits and effects of the negative ion exposure.

Curr Opin Biotechnol, 2002 Feb, 13(1), 25 - 30
Stable carbon isotopes of lipid biomarkers: analysis of metabolites and metabolic fates of environmental microorganisms; Zhang CL; Lipid biomarkers are specific compounds that are characteristic of certain groups or species of microorganisms . The use of natural or labeled carbon isotopes of lipid biomarkers has enabled a better understanding of carbon flow pathways at the molecular level . Recent advances include, but are not limited to, the elucidation of mechanisms of anaerobic methane oxidation mediated by syntrophic sulfate-reducing bacteria and Archaea, linking microbial populations with specific microbial processes or bacterial transport mechanisms in natural or contaminated environments, and elucidation of the biosynthetic pathways of cellular material.

Curr Opin Biotechnol, 2001 Dec, 12(6), 587 - 604
Biocatalytic synthesis of intermediates for the synthesis of chiral drug substances; Patel RN; There has been an increasing awareness of the enormous potential of microorganisms and enzymes for the transformation of synthetic chemicals with high chemo-, regio- and enantioselectivity . Chiral intermediates and fine chemicals are in high demand, both from the pharmaceutical and agrochemical industries, for the preparation of bulk drug substances and agricultural products . Biocatalytic processes have been described for the synthesis of chiral intermediates for beta3- and beta2-receptor agonists, antihypertensive drugs, antiviral agents, melatonin receptor agonists, anticholesterol and anticancer drugs, and drugs to treat Alzheimer's disease.

J Food Prot, 2002 Feb, 65(2), 411 - 4
Decontaminative effect of frozen acidic electrolyzed water on lettuce; Koseki S et al.; We investigated the effects of frozen acidic electrolyzed water (AcEW) on lettuce during storage in a styrene-foam container . The lettuce was kept at 2 to 3 degrees C for 24 h . Populations of aerobic bacteria associated with lettuce packed in frozen AcEW were reduced by 1.5 log CFU/g after storage for 24 h . With frozen tap water, no microorganism populations tested in this study were reduced . A frozen mixture of AcEW and alkaline electrolyzed water (AlEW) also failed to reduce populations of microorganisms associated with lettuce . Although chlorine gas was produced by frozen AcEW, it was not produced by the AcEW-AlEW mixture . This result indicates that the main factor in the decontaminative effect of frozen AcEW was the production of chlorine gas . Accordingly, low-temperature storage and decontamination could be achieved simultaneously with frozen AcEW during distribution.

Chem Pharm Bull (Tokyo), 2002 Feb, 50(2), 175 - 8
Purification and characterization of ginsenoside-alpha-L-rhamnosidase; Yu H et al.; In this paper the ginsenoside-alpha-(1-->2)-L-rhamnosidase from microorganisms was purified and characterized . The enzyme hydrolyzed the 6-C, alpha-(1-->2)-L-rhamnoside of 20(S) and 20(R)-ginsenoside Rg2 to produce the 20(S) and 20(R)-ginsenoside Rh1, but hardly hydrolyzed the alpha-rhamnoside of pNPR . The enzyme molecular weight was about 53 kDa . The optimum temperature of enzyme reaction was 40 degrees C, and the optimum pH was 5.

Environ Toxicol, 2002 Feb, 17(1), 49 - 62
The fate of diesel hydrocarbons in soils and their effect on the germination of perennial ryegrass; Siddiqui S et al.; Hydrocarbon contamination in soils may be toxic to plants and soil microorganisms and act as a source of groundwater contamination . The objective of this study was to evaluate the fate of diesel in soils with or without added nutrients . The soils examined either had or had not a previous history of hydrocarbon contamination . Particular aspects examined were soil respiration, changes in microbial population, breakdown of diesel hydrocarbons, and phytotoxicity to the germination of perennial ryegrass . Soil respiration was measured as evolved CO2 . Bacterial population was determined as colony forming units in dilution plates and fungal activity was measured as hyphal length . The fate of individual hydrocarbons was determined by gas chromatography-mass spectrometry after extraction with dichloromethane . When diesel was added to soil with no previous history of hydrocarbon contamination at rates up to 50 mg/g, the respiration response showed a lag phase of 6 days and maximum respiration occurred at day 11 . The lag phase was 2 days and maximum respiration occurred at day 3 in soil with a previous history of hydrocarbon contamination . After the peak, respiration decreased up to about 20 days in both soils . Thereafter, respiration become more or less constant but substantially greater than the control . N and P addition along with diesel did not reduce the lag phase but increased the respiration over the first 20 days of incubation . Diesel addition with or without N and P increased the bacterial population 10- to 100-fold but fungal hyphal length did not increase . Diesel addition at a rate of 136 mg/g did not increase the microbial population . Removal of inhibition to germination of perennial ryegrass was linked to the decomposition of nC10 and nC11 hydrocarbons and took from 11 to 30 days at diesel additions up to 50 mg/g depending on the soil . Inhibition to germination of perennial ryegrass persisted to more than 24 weeks at the 136 mg/g of diesel addition.

Bioinformatics, 2002 Feb, 18(2), 351 - 61
Exploring the pathway structure of metabolism: decomposition into subnetworks and application to Mycoplasma pneumoniae; Schuster S et al.; MOTIVATION: Reconstructing and analyzing the metabolic map of microorganisms is an important challenge in bioinformatics . Pathway analysis of large metabolic networks meets with the problem of combinatorial explosion of pathways . Therefore, appropriate algorithms for an automated decomposition of these networks into smaller subsystems are needed . RESULTS: A decomposition algorithm for metabolic networks based on the local connectivity of metabolites is presented . Interrelations of this algorithm with alternative methods proposed in the literature and the theory of small world networks are discussed . The applicability of our method is illustrated by an analysis of the metabolism of Mycoplasma pneumoniae, which is an organism of considerable medical interest . The decomposition gives rise to 19 subnetworks . Three of these are here discussed in biochemical terms: arginine degradation, the tetrahydrofolate system, and nucleotide metabolism . The interrelations of pathway analysis of biochemical networks with Petri net theory are outlined.

Cryobiology, 2001 Sep, 43(2), 124 - 32
Anhydrobiotic engineering of bacterial and mammalian cells: is intracellular trehalose sufficient?
Tunnacliffe A, Garcia de Castro A, Manzanera M.
Anhydrobiotic engineering aims to confer a high degree of desiccation tolerance on otherwise sensitive living organisms and cells by adopting the strategies of anhydrobiosis . Nonreducing disaccharides such as trehalose and sucrose are thought to play a pivotal role in resistance to desiccation stress in many microorganisms, invertebrates, and plants, and in vitro trehalose is known to confer stability on dried biomolecules and biomembranes . We have therefore tested the hypothesis that intracellular trehalose (or a similar molecule) may be not only necessary for anhydrobiosis but also sufficient . High concentrations of trehalose were produced in bacteria by osmotic preconditioning, and in mammalian cells by genetic engineering, but in neither system was desiccation tolerance similar to that seen in anhydrobiotic organisms, suggesting that trehalose alone is not sufficient for anhydrobiosis . In Escherichia coli such desiccation tolerance was achievable, but only when bacteria were dried in the presence of both extracellular trehalose and intracellular trehalose . In mouse L cells, improved osmotolerance was observed with up to 100 mM intracellular trehalose, but desiccation was invariably lethal even with extracellular trehalose present . We conclude that anhydrobiotic engineering of at least some microorganisms is achievable with present technology, but that further advances are needed for similar desiccation tolerance of mammalian cells .

J Environ Sci Health A Tox Hazard Subst Environ Eng, 2002, 37(2), 213 - 36
Tetrachloroethylene (TCE) removal during anaerobic granulation in an upflow anaerobic sludge blanket (UASB) reactor; Sponza DT; The anaerobic biotransformation of TCE was investigated during the granulation process by reducing the HRT and increasing the chemical oxygen demand (COD) and TCE loadings in a 21 laboratory-scale upflow anaerobic sludge blanket (UASB) reactor . Anaerobic unacclimated sludge and glucose were used as seed and primary substrate, respectively . Initial granules were developed after 1.5 months of start-up, which grew at an accelerated pace for 7 months and then became fully grown . The effect of operational parameters such as influent TCE concentrations, COD and TCE loading, food to microorganism (F/M) ratio and specific methanogenic activity (SMA) were also considered during granulation . The granular sludge cultivated had a maximum diameter of 2.5 mm and SMA of 1.32 gCOD (gTSS day)(-1) while 94% COD and 90% TCE removal efficiencies were achieved when the reactor was operating at loading rates as high as 160 mg TCE (1 day)(-1) and 14 gCOD (1 day)(-1), respectively after 230 days of continuos operation . This corresponds to HRT of 0.28 day and F/M ratio of 0.60 gCOD (gTSS day)(-1) . Kinetic coefficients of k (maximum specific substrate utilization rate), Ks (half velocity coefficient), Y (growth yield coefficient) and b (decay coefficient) were determined to be 2.38 mgCOD (mgTSS day)(-1), 108 mgCOD 1(-1), 0.17 mgTSS (mgCOD)(-1) and 0.015 day(-1), respectively during TCE biotransformation based on glucose-COD during granulation.

Immunobiology, 2001 Dec, 204(5), 558 - 71
Complex encounters at the macrophage-mycobacterium interface: studies on the role of the mannose receptor and CD14 in experimental infection models with Mycobacterium avium; Reiling N et al.; The initial interactions between mycobacterial cell wall components and receptor structures on the surface of macrophages may be critical in determining the outcome of infection . They may trigger the ingestion and digestion of microorganisms, but they may also promote the intracellular persistence and growth of mycobacteria . Using Mycobacterium avium as a model system, three approaches of different complexities were used to analyse some structural features and some functional consequences of M . avium interacting with the macrophage mannose receptor or CD14, a pattern recognition receptor . Binding specificities of a recombinant, truncated extracellular portion of the mannose receptor were assayed in a novel ELISA-formatted system using viable M . avium cells as ligands . Infection with M . avium strains differing in their virulence were performed in murine bone marrow-derived macrophages and in mice with a targeted deletion of the CD14 gene . These parallel and converging approaches not only help define the molecular basis for understanding early events in the pathogenesis of mycobacterial infections, but are also necessary to ultimately determine the relevance of in vitro findings in the context of actual manifestations of disease in vivo.

Sci Total Environ, 2002 Feb 4, 284(1-3), 227 - 35
Selenium in soil and endemic diseases in China; Tan J et al.; Selenium is an essential element for humans, animals and some species of microorganisms . The biological function of selenium shows dual characteristics . The selenium content range between toxic and deficient concentration is very narrow . The present paper discusses the geographical distribution of two forms (total and water-soluble) of selenium in topsoil (plough layer for cultivated soils, eluvial horizon for natural soils) and evaluates its relationship with some human health problems in China . Topsoil samples, 354 in total, including 156 natural and 198 cultivated soils of 21 main soil types were collected . The total Se concentration in soil samples was determined with DAN (di-aminonaphthalene)-fluorescence spectrophotometer method . Soil water-soluble Se concentration was determined with the same method after extraction with water (water/soil = 5:1) . The results showed that the geometric and arithmetic means of total Se concentration in soil, for all samples, were 0.173 mg/kg and 0.239 mg/kg, respectively, with the lowest value being 0.022 mg/kg and the highest being 3.806 mg/kg . For the cultivated soil, the geometric mean of total Se was 0.188 mg/kg, its arithmetic mean was 0.269 mg/kg and higher than those in the natural soil, 0.154 mg/kg and 0.206 mg/kg, respectively . The geometric and arithmetic means of water-soluble Se in soil for all the samples were 4.0 and 6.4 microg/kg, the lowest 0.6 microg/kg and the highest value being 109.4 microg/kg . For the cultivated soils, the average concentration of water-soluble Se was 4.3 microg/kg, similar to that of natural soil, they are and 4.4 microg/kg by geometric mean . Two sequences of the soil types, arranged separately in the concentration of total Se and water-soluble Se, are different and this demonstrates that the proportions of the two forms of selenium existing in various soils are different . The percentages of water-soluble Se to total Se in different types of soils varied from 1.07 to 6.69% . However, generally the laterite and other subtropic soil still have relatively high absolute water-soluble Se contents because of their higher total Se contents . A very significant correlation between total Se and water-soluble Se has been found in cultivated soil with a correlation coefficient of 0.58 (P < 0.01) . The relationships between soil Se and human endemic diseases Keshan disease, Kashin-Back diseases and selenosis have been discussed . The reference criteria for evaluating Se deficiency and Se excess in soil were suggested.

Roum Arch Microbiol Immunol, 2000 Jan-Jun, 59(1-2), 43 - 53
The migratory and phagocytic activity of polymorphonuclear leukocytes in rheumatoid arthritis and osteoarthritis patients; Dolganiuc A et al.; Polymorphonuclear neutrophils (PMN) play a central role in the elimination of most extracellular pathogenic microorganisms and any impairment of their functions therefore predisposes to defect immune defence . We investigated the migratory and phagocytic functions of the PMNs isolated from peripheral blood and synovial fluid of patients with rheumatoid arthritis (RA) and osteoarthritis (OA) . The results suggest that in RA the number and the migratory but not phagocytic capacity of synovial fluid (SF) neutrophils were enhanced, while in OA they were significantly decreased in synovial fluid cells comparatively with peripheral blood (PB) . The migratory function of both PB and SF cells from RA patients was increased comparatively with that of the cells from OA patients . We found the different abnormal functions in synovial fluid neutrophils from RA and OA patients . These results may help to elucidate the underlying mechanism which leads to severe joint destruction and different susceptibility to infectious diseases in patients with rheumatic disorders.

Respirology, 2001 Dec, 6(4), 311 - 5
Clinical significance of potential pathogenic microorganisms of sputum in patients with pulmonary tuberculosis; Shirai M et al.; BACKGROUND: Bacterial culture of sputum is frequently positive in patients with pulmonary tuberculosis (TB) . However, it remains to be clarified whether detection of potential pathogenic microorganisms (PPM) in sputum represents bacterial infection or only colonization of the respiratory tract . In the present study, we investigated the clinical significance of PPM in patients with pulmonary TB . METHODOLOGY: Sputum culture for PPM was studied in 174 pulmonary TB patients (117 males and 57 females, mean age of 71 years) on the day of admission . Ninety-seven patients (63 males and 34 females, mean age of 67.1 years) also underwent transtracheal aspirates (TTA) . Quantitative culture of sputum and TTA was performed to distinguish infection from colonization . The results were evaluated as follows: more than 106 or 105 c.f.u./mL indicated infection in the case of sputum or TTA samples, respectively . We also compared various clinical parameters between patients with PPM and patients with non-potential pathogenic microorganisms (non-PPM) . RESULTS: Potential pathogenic microorganisms were positive in 44% and 24% of sputum and TTA specimens, respectively . However, quantitative culture for PPM showed positive findings in only 11% and 3% of sputum and TTA specimens, respectively . A comparative study of the clinical features revealed that PPM patients had a lower bodyweight and lower serum albumin levels than non-PPM patients . A fatal outcome was also more common in PPM patients than in non-PPM patients . Logistic regression analysis further confirmed that PPM clearly contributed to a fatal outcome in addition to the previously established parameters including age, performance status, haemoglobin, albumin and radiographic disease extent . CONCLUSION: Although sputum PPM represent only colonization of the upper respiratory tract in TB patients, they are associated with a poor prognosis.

J Eur Acad Dermatol Venereol, 2001 Nov, 15(6), 532 - 40
Acne inversa (alias hidradenitis suppurativa); Jansen I et al.; Acne inversa is a recurrent, suppurative disease manifested by abscesses, fistulas, and scarring . Once considered to be a disease of the apocrine glands, it is actually a defect of follicular epithelium . Thus, the term hidradenitis suppurativa is a misnomer and should be abandoned . In cases of familial acne inversa, the pattern of transmission and number of affected individuals are consistent with autosomal dominant inheritance . Aetiological factors such as hyperandrogenism, obesity, smoking and chemical irritants are not consistently associated with the affection . Bacterial involvement is not a primary event in acne inversa, but is secondary to the disease process . Potential complications include dermal contraction, local or systemic infection due to the spread of microorganisms, systemic amyloidosis, arthropathy, and squamous cell carcinoma . As spontaneous resolution is rare and progressive disability is the rule, early definitive surgical intervention is advisable . The surgical procedure of choice in most cases is wide local excision and healing by secondary intention . Pharmacotherapeutic drugs, including synthetic retinoids and antiandrogens, do not prevent progression of the disease.

Space Med Med Eng (Beijing), 2001 Oct, 14(5), 360 - 3
{Feasibility of the use of degraded inedible biomass of plants as a nutrient liquid for hydroponic cultivation}; Guo SS et al.; Objective . To demonstrate that the recycled liquid, which originated from lettuce inedible biomass degraded by fixed microorganism (correction of microorgannism) and enzyme, can be used as a nutrient solution for lettuce hydroponic cultivation . Method . After biologically degrading the weighted, oven-dried and milled leaves and roots of lettuce in a biological reactor under aerobic condition, the original effluent and its supplemented effluent were used as nutrients for lettuce hydroponic cultivation . Result . The average dried weight (ADW) of lettuce from the original effluent group was approximately half of that from the control group, and the ADW from supplemented effluent group was about equal to that from the control group; some qualities of the lettuce such as a relatively lower content of NO3- from both the original effluent group and the supplemented effluent one improved, and some of those such as a relatively higher content of NO2- dropped . Conclusion . The biologically-degraded effluent was able to be used as nutrient solution for lettuce hydroponic cultivation, although the effects of the inorganic ion-supplemented effluent were much better; the plants of lettuce from the biologically-degraded effluent were safely edible.

Biopolymers, 2002, 67(1), 1 - 9
Medical applications of Raman spectroscopy: from proof of principle to clinical implementation; Choo-Smith LP et al.; Raman spectroscopy has recently been applied ex vivo and in vivo to address various biomedical issues such as the early detection of cancers, monitoring of the effect of various agents on the skin, determination of atherosclerotic plaque composition, and rapid identification of pathogenic microorganisms . This leap in the number of applications and the number of groups active in this field has been facilitated by several technological advancements in lasers, CCD detectors, and fiber-optic probes . However, most of the studies are still at the proof of concept stage . We present a discussion on the status of the field today, as well as the problems and issues that still need to be resolved to bring this technology to hospital settings (i.e., the medical laboratory, surgical suites, or clinics) . Taken from the viewpoint of clinicians and medical analysts, the potential of Raman spectroscopic techniques as new tools for biomedical applications is discussed and a path is proposed for the clinical implementation of these techniques .

Med Clin (Barc), 2002 Feb 9, 118(4), 130 - 4
{Pulmonary function and quality of life in relation to bronchial colonization in adults with bronchiectasis not caused by cystic fibrosis}; Hernandez C et al.; BACKGROUND: We aimed at assessing the pulmonary function and life quality of patients with bronchiectasis not caused by cystic fibrosis (CF) on the basis of the presence or absence of colonization (with Pseudomonas or other microorganisms) . PATIENTS AND METHOD: Prospective, randomised control-case study of patients with bronchiectasis who came to the Pneumology area of the Hospital Universitario de Canarias between January 1999 and December 2000 in a stable clinic situation . Patients must had no antibiotic therapy over last six weeks before the study . Patients with CF and patients who had an acute respiratory disease were excluded . We obtained two sputum samples for culture with an interval of six weeks between both . We determined the expiratory flow in the first second (FEV1), the forced vital capacity (FVC) and arterial gases . Quality of life was measured by the St . George respiratory questionnaire . RESULTS: We included 70 patients, 25 males (35%) and 45 females (64%), with a mean (SD) age of 56 (17) years . There were 14 patients who had Pseudomonas (Ps-group), 10 who had other microorganisms (another-group) and 46 patients who had no microorganism in sputum culture (no-group) . The pulmonary function of the no-group (FEV<SUB<1%: 73 {24} and FVC%: 79 {21}) was better than that of the Ps-group (FEV1%: 47 {27}; p = 0.00; FVC%: 61 {28}; p = 0.04) . The quality of life of the no-group (total score (TS): 33.2 {18.9}) was better than that of the Ps-group (TS: 54 {23.2}; p = 0.00) and that of the another-group (TS: 52.2 {20.4}; p = 0.02) . CONCLUSIONS: Non CF patients whose bronchiectasis are colonized with Pseudomonas have worse pulmonary function and quality of life than uncolonized ones . Moreover, patients having microorganisms other than Pseudomonas have a worse quality of life than those without microorganisms.

Biochemistry, 2002 Feb 19, 41(7), 2198 - 208
Structure of Escherichia coli aminodeoxychorismate synthase: architectural conservation and diversity in chorismate-utilizing enzymes; Parsons JF et al.; Aminodeoxychorismate synthase is part of a heterodimeric complex that catalyzes the two-step biosynthesis of 4-amino-4-deoxychorismate, a precursor of p-aminobenzoate and folate in microorganisms . In the first step, a glutamine amidotransferase encoded by the pabA gene generates ammonia as a substrate that, along with chorismate, is used in the second step, catalyzed by aminodeoxychorismate synthase, the product of the pabB gene . Here we report the X-ray crystal structure of Escherichia coli PabB determined in two different crystal forms, each at 2.0 A resolution . The 453-residue monomeric PabB has a complex alpha/beta fold which is similar to that seen in the structures of homologous, oligomeric TrpE subunits of several anthranilate synthases of microbial origin . A comparison of the structures of these two classes of chorismate-utilizing enzymes provides a rationale for the differences in quaternary structures seen for these enzymes, and indicates that the weak or transient association of PabB with PabA during catalysis stems at least partly from a limited interface for protein interactions . Additional analyses of the structures enabled the tentative identification of the active site of PabB, which contains a number of residues implicated from previous biochemical and genetic studies to be essential for activity . Differences in the structures determined from phosphate- and formate-grown crystals, and the location of an adventitious formate ion, suggest that conformational changes in loop regions adjacent to the active site may be needed for catalysis . A surprising finding in the structure of PabB was the presence of a tryptophan molecule deeply embedded in a binding pocket that is analogous to the regulatory site in the TrpE subunits of the anthranilate synthases . The strongly bound ligand, which cannot be dissociated without denaturation of PabB, may play a structural role in the enzyme since there is no effect of tryptophan on the enzymic synthesis of aminodeoxychorismate . Extensive sequence similarity in the tryptophan-binding pocket among several other chorismate-utilizing enzymes, including isochorismate synthase, suggests that they too may bind tryptophan for structural integrity, and corroborates early ideas on the evolution of this interesting enzyme family.

Zhonghua Yan Ke Za Zhi, 2001 Nov, 37(6), 443 - 6
{Orthokeratology induced infective corneal ulcer}; Lu L et al.; OBJECTIVE: To investigate the clinical course, treatment and outcome of infective corneal ulcer induced by orthokeratology . METHODS: Sixteen cases of infective corneal ulcer caused by orthokeratology were reported, including 7 cases of pseudomonas corneal ulcers, 8 cases of acanthamoeba keratitis and 1 case of mycotic keratitis . Smears and cultures from corneal scrapings for bacteria, fungi and amoeba were performed for all of the patients . According to the results of pathogenic microorganisms, different therapeutic approaches were given . Lamellar keratoplasty or penetrating keratoplasty was performed for 9 patients whose corneal lesions were serious and unresponsive to medical therapy . RESULTS: After medical treatment or keratoplasty, corneal infections of all the patients were controlled . The visual acuity of most cases was improved . CONCLUSIONS: The infective corneal ulcer is the most serious complication of orthokeratology . The treatment of infective corneal ulcer should be directed toward the elimination of the pathogenic microorganisms from the cornea . Keratoplasty continues to have a central role in the management of some advanced cases . Although the infections can be controlled, the vision of these patients is seriously damaged.

Trends Mol Med, 2002 Jan, 8(1), 23 - 30
Rab GTPases, intracellular traffic and disease; Seabra MC et al.; Membrane and protein traffic in the secretory and endocytic pathways is mediated by vesicular transport . Recent studies of certain key regulators of vesicular transport, the Rab GTPases, have linked Rab dysfunction to human disease . Mutations in Rab27a result in Griscelli syndrome, caused by defects in melanosome transport in melanocytes and loss of cytotoxic killing activity in Tcells . Other genetic diseases are caused by partial dysfunction of multiple Rab proteins resulting from mutations in general regulators of Rab activity; Rab escort protein-1 (choroideremia), Rab geranylgeranyl transferase (Hermansky-Pudlak syndrome) and Rab GDP dissociation inhibitor-alpha (X-linked mental retardation) . In infectious diseases caused by intracellular microorganisms, the function of endocytic Rabs is altered either as part of host defences or as part of survival strategy of the pathogen . The human genome is predicted to contain 60 RAB genes, suggesting that future work could reveal further links between Rab dysfunction and disease.

Prog Biophys Mol Biol, 2001 Nov, 77(3), 177 - 268
Structure and function of the radical enzyme ribonucleotide reductase; Eklund H et al.; Ribonucleotide reductases (RNRs) catalyze all new production in nature of deoxyribonucleotides for DNA synthesis by reducing the corresponding ribonucleotides . The reaction involves the action of a radical that is produced differently for different classes of the enzyme . Class I enzymes, which are present in eukaryotes and microorganisms, use an iron center to produce a stable tyrosyl radical that is stored in one of the subunits of the enzyme . The other classes are only present in microorganisms . Class II enzymes use cobalamin for radical generation and class III enzymes, which are found only in anaerobic organisms, use a glycyl radical . The reductase activity is in all three classes contained in enzyme subunits that have similar structures containing active site cysteines . The initiation of the reaction by removal of the 3'-hydrogen of the ribose by a transient cysteinyl radical is a common feature of the different classes of RNR . This cysteine is in all RNRs located on the tip of a finger loop inserted into the center of a special barrel structure . A wealth of structural and functional information on the class I and class III enzymes can now give detailed views on how these enzymes perform their task . The class I enzymes demonstrate a sophisticated pattern as to how the free radical is used in the reaction, in that it is only delivered to the active site at exactly the right moment . RNRs are also allosterically regulated, for which the structural molecular background is now starting to be revealed.

Dev Biol, 2002 Feb 1, 242(1), 1 - 14
Unseen forces: the influence of bacteria on animal development; McFall-Ngai MJ; The diversity of developmental programs present in animal phyla first evolved within the world's oceans, an aquatic environment teeming with an abundance of microbial life . All stages in the life histories of these early animals became adapted to microorganisms bathing their tissues, and countless examples of animal-bacterial associations have arisen as a result . Thus far, it has been difficult for biologists to design ways of determining the extent to which these associations have influenced the biology of animals, including their developmental patterns . The following review focuses on an emerging field, the goal of which is to understand the influence of bacteria on animal developmental programs . This integrative area of research is undergoing a revolution that has resulted from advances in technology and the development of suitable animal-bacterial systems for the study of these complex associations . In this contribution, the current status of the field is reviewed and the emerging research horizons are examined.

Am J Med Genet, 2002 Feb 1, 107(4), 311 - 6
Association analysis of polymorphisms at the interleukin-1 locus in essential hypertension; Lin RC et al.; Infection with microorganisms such as Helicobacter pylori and Chlamydia pneumoniae has been associated with coronary heart disease (CAD) and hypertension (HT) . Infection increases the release of pro-inflammatory cytokines, thus facilitating interactions that lead to vascular damage and other effects . We hypothesized that genetically determined differences in activity or responsiveness of cytokine(s) might contribute to HT . The interleukin-1 gene (IL1) cluster on chromosome 2q14 contains three related genes (IL1A, IL1B, and IL1RN) located within a 430-kb region . These encode IL-1alpha and IL-1beta, as well as their endogenous receptor antagonist, IL-1ra . The IL1RN gene has a penta-allelic 86-bp tandem repeat in intron 2 . Allele IL1RN* 2 is associated with a wide range of chronic inflammatory and autoimmune conditions, and its combination with the -31T variant of an IL1B C(-31)T polymorphism constitutes a pro-inflammatory haplotype that leads to vigorous IL-1beta production . We therefore tested each of these polymorphisms for association with HT . Subjects were white Anglo-Celtic residents of Sydney, Australia . Frequencies of IL1B C(-31)T genotypes CC, CT, and TT were 0.50, 0.40, and 0.10 in normotensive (NT) and 0.46, 0.46, and 0.08 in HT, respectively (chi(2) = 1.2, P = 0.55) . T allele frequency in NT (0.30) was similar to that in HT (0.31) . For the IL1RN variant, frequencies of alleles IL1RN* 1 and * 2 and combined minor alleles * 3, * 4, and * 5 were 0.61, 0.36, and 0.03 in NT and 0.54, 0.36, and 0.10 in HT, respectively (chi(2) = 11, P = 0.004) . In conclusion, no association of the IL1B C(- 31)T with HT was found, whereas combined frequency of the minor alleles of the IL1RN polymorphism was increased in the HT cohort studied .

Crit Care Med, 2002 Jan, 30(1 Supp), S1 - S11
Toll-like receptors; Lien E et al.; The ability of a host to sense invasion by pathogenic organisms and to respond appropriately to control infection is paramount to survival . In the case of sepsis and septic shock, however, an exaggerated systemic response may, in fact, contribute to the morbidity and mortality associated with overwhelming infections . The innate immune system has evolved as the first line of defense against invading microorganisms . The Toll-like receptors (TLRs) are a part of this innate immune defense, recognizing conserved patterns on microorganisms . These TLRs and their signaling pathways are represented in such diverse creatures as mammals, fruit flies, and plants . Ten members of the TLR family have been identified in humans, and several of them appear to recognize specific microbial products, including lipopolysaccharide, bacterial lipoproteins, peptidoglycan, and bacterial DNA . Signals initiated by the interaction of TLRs with specific microbial patterns direct the subsequent inflammatory response . Thus, TLR signaling represents a key component of the innate immune response to microbial infection.

Leuk Res, 2002 Apr, 26(4), 329 - 33
Mix-ups and mycoplasma: the enemies within; Drexler HG et al.; Human leukemia-lymphoma (LL) cell lines represent important tools for experimental research . Among the various problems associated with cell lines, the two most common concern contaminations: (1) cross-contamination with unrelated cells and (2) contamination with microorganisms, in particular mycoplasma . The bad news is that about one-third of the cell lines are either cross-contaminated or mycoplasma-infected or both . The good news is that there are means to recognize and overcome these problems . In cases where, during attempts to establish new LL cell lines, primary LL cultures are cross-contaminated with continuous cell lines, intended new cell lines simply cannot be established ("early" cross-contamination) . In cases of "late" cross-contamination of existing LL cell lines where the intrusive cells have a growth advantage, the original ("uncontaminated") cell lines may still be available elsewhere . DNA fingerprinting and cytogenetic analysis appear to be the most suitable approaches to detect cross-contaminations and to authenticate LL cell lines . A different but related aspect of "false" LL cell lines is the frequent misclassification of cell lines whereby the actual cell type of the cell line does not correspond to the purported model character of the cell line . Mycoplasma infection can have a multitude of effects on the eukaryotic cells which, due to the variety of infecting mycoplasma species and many other contributing parameters, cannot be predicted, rendering resulting data questionable at best . Practical procedures for the detection and elimination of mycoplasma contamination have been developed . Diagnostic and preventive strategies in order to hem the alarming increase in "false" and mycoplasma-positive LL cell lines are recommended.

Curr Issues Mol Biol, 2002 Jan, 4(1), 19 - 32
Advances towards integrated biodetection systems for environmental molecular microbiology; Chandler DP; To overcome many of the limitations associated with indirect detection methods, new techniques for the sensitive, specific, and direct detection of nucleic acids are required in order to accurately and quantitatively ascribe phenotype/function to uncultivated microorganisms . However, if advanced diagnostic and detection systems are going to be applied in environmental microbiology, future "biodetection" technologies and systems must be developed not from the point of view of the detector, but from the unique aspects of the environmental sample and the entire analytical process . This article highlights recent advances in nucleic acid-based technologies, and looks towards future advances that may address the broad needs and conditions imposed by environmental molecular microbiology.

Microbiol Immunol, 2001, 45(12), 801 - 11
Immunological properties of trehalose dimycolate (cord factor) and other mycolic acid-containing glycolipids--a review; Ryll R et al.; Mycolic acids are characteristic fatty acids of Mycobacteria and are responsible for the wax-like consistence of these microorganisms . Decades of research revealed that mycolic acid-containing glycolipids, in particular trehalose-6,6'-dimycolate (TDM, cord factor) as their best-studied representative, exert a number of immunomodifying effects . They are able to stimulate innate, early adaptive and both humoral and cellular adaptive immunity . Most functions can be associated with their ability to induce a wide range of chemokines (MCP-1, MIP-1alpha, IL-8) and cytokines (e.g., IL-12, IFN-gamma, TNF-alpha, IL-4, IL-6, IL-10) . This review tries to link well-known properties of mycolic acid-containing glycolipids, e.g., stimulation of cellular and humoral immunity, granuloma formation and anti-tumor activity, with recent findings in molecular immunology and to give an outlook on potential practical applications.

Dtsch Tierarztl Wochenschr, 2002 Jan, 109(1), 22 - 5
{Efficacy of probiotic feed additives: guidelines for the evaluation of the efficiency of microorganisms in dogs, cats, and horses}; Lahrssen M et al.; Probiotic microorganisms are frequently in use as feed additives for farm and pet animals . For admission for the common market products have to be tested according to the feed additive directive 70/524/EC . The dossier for admission has to comprise data of efficiency as laid down in the directive 87/153/EC . During the last years it became obvious after the evaluation of several dossiers, that no definitive criteria for the assessment of efficacy were available for dogs, cats and horses (84/153/EC) . Aspects like the promotion of animal production are not relevant in this context . Therefore, the European commission launched the directive for the evaluation of efficacy of microorganisms for dogs, cats and horses, that supports the evaluation of microbial feed additives and which is described in this paper.

Vestn Ross Akad Med Nauk, 2001, (11), 5 - 8
{Role of works of S . V . Prozorovskii in solving problems of persistence of wall-free forms of microorganisms}; Rakovskaia IV; S . V . Prozovsky's works are of general biological significance as they show that there can be most bacteria that are two forms having various structural arrangements: one bacteria have a wall, the others have no, such as L-form ones . The paper shows the importance of persistence of bacteria as L-forms in the pathogenesis of different infectious diseases . Mechanisms that determine the long-term persistence of Mycoplasmas in the infected organisms are explored.

Nat Genet, 2002 Mar, 30(3), 311 - 4 Epub 2002 Feb 11.
Pathogen stress increases somatic recombination frequency in Arabidopsis; Lucht JM et al.; Evolution is based on genetic variability and subsequent phenotypic selection . Mechanisms that modulate the rate of mutation according to environmental cues, and thus control the balance between genetic stability and flexibility, might provide a distinct evolutionary advantage . Stress-induced mutations stimulated by unfavorable environments, and possible mechanisms for their induction, have been described for several organisms, but research in this area has mainly focused on microorganisms . We have analyzed the influence of adverse environmental conditions on the genetic stability of the higher plant Arabidopsis thaliana . Here we show that a biotic stress factor-attack by the oomycete pathogen Peronospora parasitica-can stimulate somatic recombination in Arabidopsis . The same effect was observed when plant pathogen-defense mechanisms were activated by the chemicals 2,6-dichloroisonicotinic acid (INA) or benzothiadiazole (BTH), or by a mutation (cim3) . Together with previous studies of recombination induced by abiotic factors, these findings suggest that increased somatic recombination is a general stress response in plants . The increased genetic flexibility might facilitate evolutionary adaptation of plant populations to stressful environments.

Mass Spectrom Rev, 2001 Jul-Aug, 20(4), 157 - 71
Characterization of intact microorganisms by MALDI mass spectrometry; Fenselau C et al.; The application of MALDI mass spectrometry to desorb protein biomarkers from intact viruses, bacteria, fungus, and spores is the focus of this review . Instrumentation, sample collection, sample preparation, and algorithms for data analysis are summarized . Optimally these analyses should be carried out in less than five minutes . Successful applications are discussed from biotechnology, cell biology, and the pharmaceutical industry .

J Trauma, 2002 Feb, 52(2), 315 - 22
The pathomechanism of posttraumatic edema of lower limbs: I . The effect of extravasated blood, bone marrow cells, and bacterial colonization on tissues, lymphatics, and lymph nodes; Szczesny G et al.; BACKGROUND: The mechanical injury of soft tissues and bones of lower extremities is frequently followed by long-lasting edema at the site of trauma and distally . The pathomechanism of this complication remains unclear . Venous thrombosis and interruption of lymphatics are considered to be the main etiologic factors . We propose a concept that protracted healing of injured tissues and bones with involvement of the regional lymphatic system (lymphatics and nodes) is responsible for persistence of edema . The events affecting the first (scavenging) phase of healing of traumatized tissues, such as hematoma, translocation of bone marrow cells to soft tissues, and colonization by microorganisms, and in particular their effects on lymphatics and lymph nodes, were studied . METHODS: Mongrel dogs weighing 15-20 kg were used . Fresh blood or bone marrow cell (BMC) suspension was injected subcutaneously or intralymphatically into the paw . Strains of saprophyte bacteria residing on the skin surface were cultured and injected intradermally . Oil-contrast lymphography was performed before and after injections to evaluate the changes in lymphatics and nodes . Biopsy samples of paw skin, subcutaneous tissue, and regional lymph nodes (LN) were taken . The responsiveness of LN lymphocytes was studied in autologous mixed cultures with peripheral blood lymphocytes (PBL), BMC, and cultured bacteria . RESULTS: The PBL from subcutaneously injected blood were evacuated by the lymphatic route at a rate of 1-3%/6 hr . There was no thrombosis of lymphatic vessels or obstruction of LN sinusoids . The BMC evoked major inflammatory changes in both the skin and the LN . Bacteria caused local inflammation, dilatation of lymphatics, and destruction of node parenchyma . Autologous BMC and PBL stimulated LN lymphocytes in a 6-day culture . The responsiveness of lymph node lymphocytes to previously subcutaneously injected bacterial antigens was increased . CONCLUSION: The extravasated blood did not produce changes in skin, subcutaneous tissue, and lymphatics; however, it stimulated LN lymphocytes . The BMC and saprophyte bacteria caused major local and lymph node inflammatory response . All these factors may contribute to the local edema in the initial phases of healing of traumatized tissues.

Cesk Slov Oftalmol, 2001 Nov, 57(6), 381 - 6
{Penetrating intraocular injuries caused by foreign bodies of organic origin} }; Dolezalova J et al.; OBJECTIVE: To demonstrate two eyes of two patients with a penetrating injury by an organic intraocular foreign body (CNT), to outline the course of treatment and to evaluate anatomical and functional results . MATERIAL AND METHODS: During the period between May and July 1999 the authors treated at their department an 11-year-old girl and a 17-year old boy with a penetrating CNT injury of organic origin . In the girl they extracted a CNT which pierced the sclera and protruded into the vitreous body . The foreign body was extracted by means of a forceps, the site of perforation was treated by a cryosurgical procedure with a radial Silastic filling . The man was shot into the OL by a grain of pepper from an air-gun . The organic body in the vitreous body caused a violent inflammatory reaction in the vitreous body and retina . Early extraction of the CNT could not be made because of an adverse corneal finding which made surgery impossible . The progressing proliferative vitreoretinopathy (PVR) led to repeated formation of epiretinal, subretinal and cyclitic membranes which caused relapsing detachment of the retina (OS) . The foreign body was extracted during pars plana vitrectomy (PPV) by the transvitreal route using a forceps . Definite adherence of the retina was achieved during the third PPV, membranectomy and implantation of silicone oil (SO) . Final functional success was achieved by eliminating SO and by partial perforating keratoplasty . RESULTS: The CNT in the girl was histologically and parasitologically identified as a fibre of animal origin (animal hair or human eyelash) . The follow up period is six months, VOP-5/5 nat., NOT 17 torr, bulbus undisturbed . In the second patient the cultivation finding from the vitreous body was negative . Extensive PVR developed as a result of breakdown products of the organic CNT and led to relapsing OS . Three months after the last operation the bulbus is at rest, the corneal disc clear, the retina attached, VOL-3/60 s + 8.0 D steop., NOT 12 torr . CONCLUSIONS: The final results of treatment of severe devastating penetrating injuries by a CNT is determined by the preoperative condition of the ocular tissues . CNT of organic origin damage intraocular tissues by their breakdown products as well as by more frequent contamination with pathological microorganisms . Our experience provided evidence that penetrating injuries by non-infected CNT of organic origin have a favourable prognosis . But even relapsing and prolonged OS after extraction of the CNT of organic origin and endophthalmitis need not lead to loss of the eye or its function.

Ned Tijdschr Tandheelkd, 1994 May, 101(5), 209 - 12
{HIV and dentistry . Part 5 . Practice hygiene}; Sjamsoedin DA et al.; When complying a good standard hygiene protocol in the dental office the risk of cross-infection of whatever type of microorganism, including the hepatitis-B-virus and HIV, is almost eliminated.

J Biol Chem, 2002 Apr 19, 277(16), 14281 - 7 Epub 2002 Feb 05.
Duplicated binding sites for (1-->3)-beta-D-glucan in the horseshoe crab coagulation factor G: implications for a molecular basis of the pattern recognition in innate immunity; Takaki Y et al.; The horseshoe crab factor G, a heterodimeric serine protease zymogen, is activated by (1-->3)-beta-D-glucan on fungal cell walls . The activation initiates the hemolymph-clotting cascade, a critical reaction for the defense against microorganisms . In the present study, we identified the domain responsible for the glucan recognition by factor G and characterized its interaction with (1-->3)-beta-d-glucan and its derivatives . Among three domains in subunit alpha of factor G, identified as the glucan-binding domain, was the COOH-terminal xylanase Z-like domain composed of two tandem-repeating units, each of which exhibits sequence similarities to the cellulose-binding domains of bacterial xylanases . Each of the single units bound to the glucan with lower affinities, and the association constant increased two orders with the tandem-repeating structure (K(a) = 8.0 x 10(8) m(-1)) . In addition to longer glucans, (1-->3)-beta-D-glucan oligosaccharides incapable of activating factor G bound also to factor G and competitively inhibited the zymogen activation . The minimum structure required for the binding was a (1-->3)-beta-d-glucan disaccharide, indicating that conformation-dependent structures are not essential for the recognition . Therefore, increasing avidity by multivalent binding sites with low affinities to simple structures on biologically active polymers may be one of the principles that allows stable and specific recognition of pathogens by pattern recognition receptors in innate immunity.

J Agric Food Chem, 2002 Feb 13, 50(4), 767 - 70
Behaviors of D- and L-lactic acids during the brewing process of sake (Japanese rice wine); Kodama S et al.; The amounts of D- and L-lactic acids during the brewing process of sake were determined by capillary electrophoresis using 2-hydroxypropyl-beta-cyclodextrin as a chiral selector . Because L-lactic acid, which prevents the growth of nonuseful microorganisms, is a raw material of sake, the ratio of L-lactic acid to total lactic acid is almost 1.0 at the initial stage of sake brewing . During brewing, the ratio decreased gradually and finally reached 0.39 . Yeast (Saccharomyces cerevisiae) for sake brewing produced D-lactic acid, but not L-lactic acid in a culture medium . These results suggest that the decrease in the ratio of L-lactic acid to total lactic acid during sake brewing resulted in D-lactic acid production by yeast . The ratios in 18 brands of sake obtained commercially ranged from 0.23 to 0.78 . The levels of D-lactic acid in sake (140-274 mg/L) were in a narrower range than those of L-lactic acid (61-461 mg/L) . Although the D-lactic acid level in sake did not correspond to total lactic acid level, the L-lactic acid level correlated well with total lactic acid level (R(2) = 0.867) . These results suggest that the ratio of L-lactic acid to total lactic acid in sake reflected the amount of L-lactic acid added at the initial stage of sake brewing.

Trends Microbiol, 2002 Feb, 10(2), 70 - 4
Acid survival of Helicobacter pylori: how does urease activity trigger cytoplasmic pH homeostasis?
Stingl K, Altendorf K, Bakker EP.
Helicobacter pylori can survive for several hours at pH 1 in the presence of urea . Under these conditions, the organism maintains its cytoplasmic pH at a value close to neutral . The role of the cytoplasmically located urease enzyme in this process is a matter of debate . We propose that cytoplasmic ammonia generated by the action of urease is protonated by H(+) ions leaking in from the acidic medium and that the NH(4)(+) formed is extruded from the cytoplasm via an as-yet-unidentified transport system . This mechanism is compared with the general mechanism of cytoplasmic pH homeostasis in microorganisms.

Chemosphere, 2002 Jan, 46(2), 187 - 96
Interactions between a polycyclic aromatic hydrocarbon mixture and the microbial communities in a natural freshwater sediment; Verrhiest GJ et al.; The toxicity of a polycyclic aromatic hydrocarbon (PAH) mixture was assessed on the indigenous microbial communities of a natural freshwater sediment . The fate and effects of the PAH mixture (phenanthrene, fluoranthene and benzo(k)fluoranthene) were studied over 28 days . Bacterial communities were described by bacterial counts (total bacteria and viable bacteria), and by some hydrolytic enzyme activities (beta-glucosidase and leucine-aminopeptidase), PAH concentrations were measured in the overlying waters and in the sediments . No effect of PAH was detected at 30 mg/kg for all bacterial parameters . At 300 mg/kg, the quantity of total bacteria and the proportion of viable bacteria markedly decreased, compared to the control (0 mg PAH/kg) . At 300 mg/kg, an increase of the beta-glucosidase activity and a decrease of the leucine-aminopeptidase activity were observed . For all treatments, the benzo(k)fluoranthene concentration in the sediment was stable over 28 days whereas, in the same time, only 3-6% of the initial concentrations of phenanthrene and fluoranthene remained . This study shows that (1) PAH induce perturbations of sediment microbial communities in terms of density and metabolism (but not always as an inhibition), (2) indigenous bacteria of sediments might be used for toxicity assessment of specific organic pollutants, (3) native microorganisms of sediment seem to have a high capacity for PAH degradation, depending on the physico-chemical properties and the bioavailability of the substance encountered.

Antonie Van Leeuwenhoek, 2001 Dec, 80(3-4), 215 - 24
Kinetic profile of the cellular lipid composition in an oleaginous Yarrowia lipolytica capable of producing a cocoa-butter substitute from industrial fats; Papanikolaou S et al.; Cell growth, lipid accumulation and cellular lipid composition of Yarrowia lipolytica growing on mixtures of industrial fats containing stearic, oleic, linoleic and palmitic acid have been studied . During growth, the strain incorporated oleic and linoleic acids more rapidly than the saturated fatty acids . Relatively high lipid accumulation (up to 0.44 g of lipids per g of dry matter) was observed when stearic acid was included in the culture medium . In contrast, substrates rich in oleic acid did not favor cellular lipid accumulation . The accumulated lipids, mainly composed of triacylglycerols (45-55% w/w), demonstrated a different total fatty acid composition compared with that of the substrate; in all cases, the microorganism showed the unusual capacity to increase its cellular stearic acid level, even if this fatty acid was not found in high concentrations in the substrate . This permitted the synthesis of interesting lipid profiles with high percentages of stearic acid and non-negligible percentages of palmitic and oleic acid, with a composition resembling that of cocoa-butter.

Biosci Biotechnol Biochem, 2001 Dec, 65(12), 2802 - 5
Purification of an aminopeptidase preferentially releasing N-terminal alanine from cucumber leaves and its identification as a plant aminopeptidase N; Yamauchi Y et al.; In this study, a highly active foliar aminopeptidase preferentially releasing N-terminal alanine from artificial substrates was purified and characterized from cucumber (Cucumis sativus L . suyo) . The enzyme had a molecular mass of 200 kDa consisting of two subunits of 95 kDa . It was a metalloprotease the pH optimum of which was 8 to 9 . It cleaved Ala-, Gly-, Met-, Ser-, Leu-, Lys-, and Arg artificial substrates . An internal amino acid sequence was similar to those of aminopeptidase N (clan MA, family M1) of microorganisms, and was very similar to that of a putative aminopeptidase N of Arabidopsis thaliana . From these results, the highly active aminopeptidase in cucumber leaves was identified to be a plant aminopepitdase N.

Biodegradation, 2001, 12(3), 201 - 7
Repeated inoculation as a strategy for the remediation of low concentrations of phenanthrene in soil; Schwartz E et al.; Phenanthrene, a polycyclic aromatic hydrocarbon, becomes increasingly unavailable to microorganisms for degradation as it ages in soil . Consequently, many bioaugmentation efforts to remediate polycyclic aromatic hydrocarbons in soil have failed . We studied the effect of repeatedly inoculating a soil with a phenanthrene-degrading Arthrobacter sp . on the mineralization kinetics of low concentrations of phenanthrene . After the first inoculation, the initial mineralization rate of 50 ng/g phenanthrene declined in a biphasic exponential pattern . By three hundred hours after inoculation, there was no difference in mineralization rates between the inoculated and uninoculated treatments even though a large fraction of the phenanthrene had not yet been mineralized . A second and third inoculation significantly increased the mineralization rate, suggesting that, though the mineralization rate declined, phenanthrene remained bioavailable . Restirring the soil, without inoculation, did not produce similar increases in mineralization rates, suggesting absence of contact between cells and phenanthrene on a larger spatial scale (>mm) is not the cause of the mineralization decline . Bacteria inoculated into soil 280 hours before the phenanthrene was added could not maintain phenanthrene degradation activity . We suggest sorption lowered bioavailability of phenanthrene below an induction threshold concentration for metabolic activity of phenanthrene-degrading bacteria.

Life Sci Space Res, 1970, 8, 33 - 8
Survival of microorganisms under simulated space conditions; Bucker H et al.; Monocellular layers of stationary phase cells of Escherichia coli B/r were exposed to high vacuum of 10(-6) Torr at different temperatures . After simultaneous irradiation with UV of 254 nm or X-rays, the survival was tested as colony forming ability . In vacuum the sensitivity to UV and X-rays was enhanced compared with the controls at atmospheric pressure . Of UV-inactivation in vacuum no photoreactivation was found.

Rev Cubana Med Trop, 2001 Jan-Apr, 53(1), 5 - 15
{Emerging viral diseases}; Guzman MG et al.; In the last few years an increasing rise of new infectious diseases or of other diseases considered to be under control has been observed . The so called emerging and reemerging diseases are those new infections that have come up in a population or those existing diseases which incidence and geographic extension are on a rapid increase . Factors such as social and economic situations, medical assistance, food production, changes in human behaviours, environmental changes, health systems deterioration, and adaptation and changes of microorganisms are related with the emergence or reemergence of a number of entities . This paper sets forth an analysis of the emergence and reemergence of viral diseases and of those factors that have had an impact on this situation.

J Clin Microbiol, 2002 Feb, 40(2), 400 - 6
Identification of Mycobacterium spp . by using a commercial 16S ribosomal DNA sequencing kit and additional sequencing libraries; Cloud JL et al.; Current methods for identification of Mycobacterium spp . rely upon time-consuming phenotypic tests, mycolic acid analysis, and narrow-spectrum nucleic acid probes . Newer approaches include PCR and sequencing technologies . We evaluated the MicroSeq 500 16S ribosomal DNA (rDNA) bacterial sequencing kit (Applied Biosystems, Foster City, Calif.) for its ability to identify Mycobacterium isolates . The kit is based on PCR and sequencing of the first 500 bp of the bacterial rRNA gene . One hundred nineteen mycobacterial isolates (94 clinical isolates and 25 reference strains) were identified using traditional phenotypic methods and the MicroSeq system in conjunction with separate databases . The sequencing system gave 87% (104 of 119) concordant results when compared with traditional phenotypic methods . An independent laboratory using a separate database analyzed the sequences of the 15 discordant samples and confirmed the results . The use of 16S rDNA sequencing technology for identification of Mycobacterium spp . provides more rapid and more accurate characterization than do phenotypic methods . The MicroSeq 500 system simplifies the sequencing process but, in its present form, requires use of additional databases such as the Ribosomal Differentiation of Medical Microorganisms (RIDOM) to precisely identify subtypes of type strains and species not currently in the MicroSeq library.

Glycobiology, 2002 Jan, 12(1), 1 - 14
The salivary mucin MG1 (MUC5B) carries a repertoire of unique oligosaccharides that is large and diverse; Thomsson KA et al.; The high-molecular-mass salivary mucin MG1, one of two major mucins produced by human salivary glands, plays an important role in oral health by coating the tooth surface and by acting as a bacterial receptor . Here this mucin was purified from the submandibular/sublingual saliva of a blood group O individual . The presence of MUC5B as the major mucin in this preparation was confirmed by amino acid analysis and its reactivity with the monoclonal antibody PAN H2 . To structurally characterize MG1 carbohydrates the O-glycans were released by reductive beta-elimination . Nuclear magnetic resonance spectroscopy of the nonfractionated mixture showed that (1) fucose was present in blood group H, Le(a), Le(x), Le(b), and Le(y) epitopes; (2) NeuAc was mainly linked alpha 2-3 to Gal or alpha 2-6 to GalNAcol; and (3) the major internal structures were core 1 and core 2 sequences . After this preliminary analysis the released oligosaccharides were separated into neutral (56%), sialylated (26%), and sulfated (19%) fractions, with an average length of 13, 17, and 41 sugar residues, respectively . Gas chromatography-mass spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of mixtures of neutral and sialylated oligosaccharides revealed at least 62 neutral and 25 sialylated oligosaccharides consisting of up to 20 monosaccharide residues . These results showed that the MG1-derived oligosaccharides were much longer than those of MG2, and only a few species were found on both molecules . Thus, these two mucins create an enormous repertoire of potential binding sites for microorganisms at one of the major portals where infectious organisms enter the body.

Ophthalmology, 2002 Feb, 109(2), 343 - 50
Possible role of the vitamin E solubilizer in topical diclofenac on matrix metalloproteinase expression in corneal melting: an analysis of postoperative keratolysis; Hargrave SL et al.; OBJECTIVE: To analyze tissue matrix metalloproteinase (MMP) expression in three patients who developed postoperative corneal melts after treatment with topical diclofenac sodium 0.1% (Falcon; Fort Worth, TX) ophthalmic solution . DESIGN: Retrospective noncomparative interventional case series with tissue analysis . MAIN OUTCOME MEASURES: Three patients were examined in this study . We report two patients from the same center with acute corneal melts after uncomplicated photorefractive keratectomy (PRK) . Prior to these cases, 1500 patients were treated at the Zale Lipshy University Laser Center for Vision with no adverse effects . All 1500 patients were treated with the same postoperative regimen of ciprofloxacin, rimexolone, and suprofen ({Profenal, (CIBA, Duluth, GA}) . The next 27 cases were treated postoperatively with ciprofloxacin and rimexolone . However, diclofenac sodium 0.1% was used instead of Profenal . A third case was also discussed . This melt occurred at another center in a postoperative cataract patient who developed cystoid macular edema after cataract extraction with intraocular lens placement . He was initially treated with diclofenac sodium 0.1% (Ciba Vision, Duluth, GA) then with diclofenac sodium 0.1% . He subsequently developed a corneal perforation requiring penetrating keratoplasty . All tissue specimens were examined by light microscopy . Microbiologic cultures and stains were also performed . Immunolocalization and in situ hybridization were performed on all keratoplasty specimens to detect expression and localization of MMPs . All patients had a complete diagnostic evaluation for systemic autoimmune diseases . RESULTS: Postoperatively, all patients developed corneal perforations requiring surgical intervention while being treated with diclofenac sodium 0.1% . Microbiologic cultures and special stains were negative for microorganisms . Induced expression of specific tissue degrading enzymes of the matrix metalloproteinase family was demonstrated within corneal epithelial cells, stromal keratocytes, and at the level of Descemet's membrane . The uniform distribution pattern of expression was not consistent with the localization expected of a repair response, suggesting the involvement of some outside agent . CONCLUSIONS: Whereas MMP expression is a normal component of repair, excessive or inappropriate MMP activity is associated with corneal keratolysis . Our study provides preliminary evidence that topical application of diclofenac sodium 0.1% may be associated with aberrant MMP expression in the cornea.

Biochim Biophys Acta, 2002 Jan 31, 1594(1), 199 - 205
Structure of the major glycolipid from Rothia dentocariosa; Pasciak M et al.; Structural studies of the major glycolipid isolated from Rothia dentocariosa were carried out by specific chemical degradation and nuclear magnetic resonance spectroscopy . The glycolipid was found to be a dimannosylacylmonoglyceride in which the carbohydrate part was the glycerol-linked dimannoside alpha-D-Manp-(1-->3)-alpha-D-Manp-(1-->3)-sn-Gro, and the internal mannose was esterified at C-6 by fatty acid residue . The other fatty acyl chain substituted the primary methylene position of glycerol . The occurrence of this glycolipid is limited to the related microorganisms . The structural characteristics can facilitate the differentiation of some genera.

Appl Environ Microbiol, 2002 Feb, 68(2), 1010 - 3
Quantitation of DNA sequences in environmental PCR products by a multiplexed, bead-based method; Spiro A et al.; A first application of a multiplexed, bead-based method is described for determining the abundances of target sequences in an environmental PCR product . Target sequences as little as 0.3% of the total amount of DNA can be quantified . Tests were conducted on 16S ribosomal DNA sequences from microorganisms collected from contaminated groundwater.

Can J Microbiol, 2001 Dec, 47(12), 1101 - 6
Isolation of an esterase-producing Trichosporon brassicae and its catalytic performance in kinetic resolution of ketoprofen; Shen D et al.; A yeast strain CGMCC 0574, identified as Trichosporon brassicae, was selected from 92 strains for its high (S) selectivity in the hydrolysis of ketoprofen ethyl ester . The effective strains of the microorganisms were isolated from soil samples with the ester as the sole carbon source . The ethyl ester proved to be the best substrate for resolution of ketoprofen among several ketoprofen esters examined . The resting cells of CGMCC 0574 could catalyze the hydrolysis of ketoprofen ethyl ester with an enantiomeric ratio of 44.9, giving (S)-ketoprofen an enantiomeric excess of 91.5% at 42% conversion.

J Biol Chem, 2002 Apr 19, 277(16), 13548 - 55 Epub 2002 Jan 30.
Uniport of NH4+ by the root hair plasma membrane ammonium transporter LeAMT1;1; Ludewig U et al.; The transport of ammonium/ammonia is a key process for the acquisition and metabolism of nitrogen . Ammonium transport is mediated by the AMT/MEP/Rh family of membrane proteins which are found in microorganisms, plants, and animals, including the Rhesus blood group antigens in humans . Although ammonium transporters from all kingdoms have been functionally expressed and partially characterized, the transport mechanism, as well as the identity of the true substrate (NH(4+) or NH(3)) remains unclear . Here we describe the functional expression and characterization of LeAMT1;1, a root hair ammonium transporter from tomato (Lycopersicon esculentum) in Xenopus oocytes . Micromolar concentrations of external ammonium were found to induce concentration- and voltage-dependent inward currents in oocytes injected with LeAMT1;1 cRNA, but not in water-injected control oocytes . The NH(4+)-induced currents were more than 3-fold larger than methylammonium currents and were not subject to inhibition by Na(+) or K(+) . The voltage dependence of the affinity of LeAMT1;1 toward its substrate strongly suggests that charged NH(4+), rather than NH(3), is the true transport substrate . Furthermore, ammonium transport was independent of the external proton concentration between pH 5.5 and pH 8.5 . LeAMT1;1 is concluded to mediate potential-driven NH(4+) uptake and retrieval depending on root membrane potential and NH(4+) concentration gradient.

Am Fam Physician, 2002 Jan 15, 65(2), 241 - 8
Genitourinary infections and their association with preterm labor; Cram LF et al.; Genitourinary tract infections are one cause of preterm delivery . Prematurity is one of the leading causes of perinatal mortality in the United States . Uterine contractions may be induced by cytokines and prostaglandins, which are released by microorganisms . Asymptomatic bacteriuria, gonococcal cervicitis and bacterial vaginosis are strongly associated with preterm delivery . The role of Chlamydia trachomatis, Trichomonas vaginalis and Ureaplasma urealyticum is less clear . By adopting a rational approach to the diagnosis and treatment of genitourinary infections, family physicians can substantially decrease a patient's risk of preterm delivery.

J Chem Ecol, 2001 Nov, 27(11), 2219 - 31
Volatiles from potential hosts of Rhopalicus tutela a bark beetle parasitoid; Pettersson EM; Host location cues for parasitic wasps that attack bark beetle larvae concealed under the bark of spruce trees were analyzed by collecting odor samples from entrance holes into the bark beetle galleries, isolated larvae, and pupal chambers with or without bark beetle larvae . Odor samples were collected by dynamic headspace adsorptions on Porapak Q or static adsorptions by using solid-phase microextraction (SPME) with Carbowax-divinylbenzene as the adsorbing phase . Samples were analyzed by coupled gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometry (GC-MS) . The antennae of Rhopalicus tutela females responded primarily to oxygenated monoterpenes that are typical for damaged host trees . These compounds are attractive to bark beetle parasitoids in long-range host location, suggesting that they are used in both long- and short-range host location . No differences could be detected between samples collected from pupal chambers with or without mature larvae . Larvae outside pupal chambers emitted low quantities of the same compounds present in empty pupal chambers . The data support the hypothesis that volatiles used by host foraging parasitoids arise from the interaction between introduced microorganisms and the bark and/or vascular tissue of the host tree rather than from the bark beetle larvae.

Antonie Van Leeuwenhoek, 2001 Sep, 79(3-4), 261 - 7
Assessment of chitin decomposer diversity within an upland grassland; Krsek M et al.; The breakdown of chitin within an acidic upland grassland was studied . The aim was to provide a molecular characterisation of microorganisms involved in chitin degradation in the soil using soil microcosms and buried litter bags containing chitin . The investigation involved an examination of the effects of liming on the microbial communities within the soil and their chitinolytic activity . Microcosm experiments were designed to study the influence of lime and chitin enrichment on the grassland soil bacterial community ex situ under controlled environmental conditions . Bacterial and actinomycete counts were determined and total community DNA was extracted from the microcosms and from chitin bags buried at the experimental site . PCR based on specific 16S rRNA target sequences provided products for DGGE analysis to determine the structure of bacterial and actinomycete communities . Chitinase activity was assessed spectrophotometrically using chitin labelled with remazol brilliant violet . Both liming and chitin amendment increased bacterial and actinomycete viable counts and the chitinase activity . DGGE band patterns confirmed changes in bacterial populations under the influence of both treatments . PCR products amplified from DNA isolated from chitin bags were cloned and sequenced . Only a few matched known species but a prominent coloniser of chitin proved to be Stenotrophomonas maltophilia.

J Crit Care, 2001 Dec, 16(4), 182 - 7
Suppression of Helicobacter pylori infection during intensive care stay: related to stress ulcer bleeding incidence?
van der Voort PH, van der Hulst RW, Zandstra DF, Geraedts AA, van der Ende A, Tytgat GN.
PURPOSE: To assess the prevalence of active Helicobacter pylori infection in patients admitted to the intensive care unit, to determine the effect of selective gut decontamination on the persistence of this organism, and to explore the possible relationship between H . pylori infection and stress ulcer bleeding incidence . MATERIALS AND METHODS: We determined in a prospective observational study of 300 consecutive, mechanically ventilated patients the activity of H . pylori infection and the incidence of stress ulcer-related upper gastrointestinal bleeding over time . H . pylori infection was detected by Laser-Assisted Ratio Analyzer (LARA)- (13)C-urea breath test (Alimenterics, Inc., NJ) and serology . Stress ulcer prophylaxis was not prescribed . Endoscopy was performed in cases of upper gastrointestinal bleeding . RESULTS: The prevalence of active H . pylori infection on admission was 38% as detected by urea breath test, and declined to 8% on the third day, and to 0% on the seventh day after admission as a result of antibiotic treatment . Stress ulcer-related upper gastrointestinal bleeding occurred in 1.0% (3 of 300) of the patients; none were infected with H . pylori on admission or at the time of bleeding . CONCLUSIONS: H . pylori infection monitored by LARA- (13)C-urea breath test was rapidly suppressed during intensive care treatment, which can be explained by the routine use of antibiotics for gut decontamination.The low incidence of stress ulcer-related bleeding might be related to the prevention of H . pylori-associated stress lesions by effective suppression of this microorganism, but further studies are warranted to test this hypothesis .

Curr Microbiol, 2002 Feb, 44(2), 88 - 93
Nickel affects activity more than expression of hydrogenase protein in Frankia; Mattsson U et al.; The effects of nickel on hydrogen uptake and the post-translational processing of the large subunit of the hydrogenase protein in three Frankia strains (one isolated from an Alnus-Frankia symbiosis and two from Casuarina-Frankia associations) were investigated . All three strains responded to the addition of nickel with an increase in hydrogen uptake . Additional nickel did not affect nitrogenase activity, however evolved hydrogen was detected in Frankia KB5 in the absence of additional nickel, indicating that hydrogenase was not active . No increase in the processing rate of the hydrogenase large subunit was found with increasing nickel concentrations for any of the strains, indicating that the strategy for regulating hydrogenase in Frankia is different from that in other microorganisms.

Protein Expr Purif, 2002 Feb, 24(1), 99 - 104
Overproduction, purification, and characterization of recombinant aspartate semialdehyde dehydrogenase from Arabidopsis thaliana; Paris S et al.; In plant and microorganisms, aspartate semialdehyde dehydrogenase (ASDH) produces the branch point intermediate between the lysine and threonine/methionine pathways . In this study, we report the first cDNA cloning, purification, and characterization of a plant ASDH . The Arabidopsis thaliana ASDH is an homodimeric enzyme composed of subunits of 36 kDa . The plant enzyme exhibited a specific activity of 26 micromol NADPH oxidized min(-1) mg(-1) of protein with a K(M) value for NADPH of 92 microM . ASDH showed cooperative behavior for aspartyl phosphate with a K(0.5) value of 37 microM .

J Theor Biol, 2002 Jan 21, 214(2), 305 - 19
INDISIM, an individual-based discrete simulation model to study bacterial cultures; Ginovart M et al.; An individual-based model has been developed and designed to simulate the growth and behaviour of bacterial colonies . The simulator is called INDISIM, which stands for INDividual DIScrete SIMulations . INDISIM is discrete in space and time, and controls a group of bacterial cells at each time step, using a set of random, time-dependent variables for each bacterium . These variables are used to characterize its position in space, biomass, state in the cellular reproduction cycle as well as other individual properties . The space where the bacterial colony evolves is also discrete . A physical lattice is introduced, subject to the appropriate boundary conditions . The lattice is subdivided into spatial cells, also defined by a set of random, time-dependent variables . These variables may include concentrations of different types of particles, nutrients, reaction products and residual products . Random variables are used to characterize the individual bacterium and the individual particle, as well as the updating of individual rules . Thus, the simulations are stochastic rather than deterministic . The whole set of variables, those that characterize the bacterial population and the environment where they evolve, enables the simulator to study the behaviour of each microorganism-such as its motion, uptake, metabolism, and viability-according to given rules suited for the system under study . These rules require the input of only a few parameters . Once this information is inputted, INDISIM simulates the behaviour of the system providing insights into the global properties of the system from the assumptions made on the properties of the individual bacteria . The relation between microscopic and global properties of the bacterial colony is obtained by using statistical averaging . In this work INDISIM has been used to study (a) biomass distributions, (b) the relationship between the rate of growth of a bacterial colony and the nutrient concentration and temperature, and (c) metabolic oscillations in batch bacterial colonies . The simulation results are found to be in very good qualitative agreement with available experimental data, and provide useful insights into the mechanisms involved in each case .

J Invertebr Pathol, 2001 Aug, 78(2), 87 - 97
Parasites and pathologic conditions of the cockle Cerastoderma edule populations of the coast of Galicia (NW Spain); Carballal MJ et al.; A survey of pathological conditions affecting cockle populations of the most economically important natural beds of Galician estuaries in NW Spain was performed . Samples of 30 adult cockles were collected from each of 34 natural beds in the spring of 1999 and processed by histological techniques . Disseminated neoplasia were seen in samples from most of the natural beds, in some cases with a high prevalence . The gregarine Nematopsis sp., larval trematode stages, and branchial extracellular large cysts enclosing bacteria-like microorganisms were the most prevalent parasites . Paravortex cardii, intracellular colonies of rickettsiae-like organisms in digestive and gill epithelium, Pseudoklossia sp . coccidians, Trichodina sp., and other ciliates were frequently seen in the samples . Copepods in gills and intestine and unidentified gregarines in intestine epithelia and surrounding connective tissue were less prevalent and were observed in samples of some natural beds . Large foci of heavy hemocytic infiltration were detected in a few sites only . Cysts of Steinhausia sp . and plasmodia and spores of a haplosporidian were seen in cockles from two localities . Inflammation was frequently observed in the samples . Some of the parasites and pathological conditions could be associated with mortality .

Anal Biochem, 2002 Feb 1, 301(1), 82 - 90
Long-read pyrosequencing using pure 2'-deoxyadenosine-5'-O'-(1-thiotriphosphate) Sp-isomer; Gharizadeh B et al.; Pyrosequencing, a nonelectrophoretic DNA sequencing method that uses a luciferase-based enzymatic system to monitor DNA synthesis in real time, has so far been limited to sequencing of short stretches of DNA . To increase the signal-to-noise ratio in pyrosequencing the natural dATP was replaced by dATPalphaS (M . Ronaghi et al., 1996, Anal . Biochem . 242, 84-89) . The applied dATPalphaS was a mixture of two isomers (Sp and Rp) . We show here that by the introduction of pure 2'-deoxyadenosine-5'-O'-(1-thiotriphosphate) Sp-isomer in pyrosequencing substantial longer reads could be obtained . The pure Sp-isomer allowed lower nucleotide concentration to be used and improved the possibility to read through poly(T) regions . In general, a doubling of the read length could be obtained by the use of pure Sp-isomer . Pyrosequencing data for 50 to 100 bases could be generated on different types of template . The longer read will enable numerous new applications, such as identification and typing of medically important microorganisms as well as resequencing of DNA fragments for mutation screening and clone checking.

J Food Prot, 2002 Jan, 65(1), 185 - 91
Survival of Pantoea agglomerans strain CPA-2 in a spray-drying process; Costa E et al.; Spray drying could be a suitable method for preserving microorganisms, as it allows large quantities of cultures to be dried at low cost . The aims of this paper were to evaluate the effects of spray-drying conditions on survival of the biocontrol agent Pantoea agglomerans CPA-2, which has shown antifungal activity against Penicillium expansum and Penicillium digitatum on citrus fruits . Various compounds cited in the bibliography as carriers were tested in our spray drying, and some salts (MgSO4, K2SO4 . and Na2CO3) and dairy products (lactoserum or nonfat skimmed milk {NFSM}) showed the best results in terms of recovered powder . Outlet temperature had more influence on the death of bacteria than inlet temperature . P . agglomerans was heat sensitive, and the activation energy was around 6 kcal/mol K when MgSO4 (10%) or NFSM (10%) were used as carriers and only 3 kcal/mol K when the combination of MgSO4 (10%) and NFSM (10%) was used . The highest powder recovery was obtained when NFSM was used as the rehydration medium . Although the percentage of powder recovery was not high (around 50%) and viability was low, the results suggest that with bigger spray dryers, we could expect a lower outlet temperature and probably an increased viability . Further research into spray-dryer design is needed in order to demonstrate this.

J Biosci, 2001 Dec, 26(5), 667 - 83
Stress-induced evolution and the biosafety of genetically modified microorganisms released into the environment; Velkov VV; This article is focused on the problems of reduction of the risk associated with the deliberate release of genetically modified microorganisms (GMMs) into the environment . Special attention is given to overview the most probable physiological and genetic processes which could be induced in the released GMMs by adverse environmental conditions, namely: (i) activation of quorum sensing and the functions associated with it, (ii) entering into a state of general resistance, (iii) activation of adaptive mutagenesis, adaptive amplifications and transpositions and (iv) stimulation of inter-species gene transfer . To reduce the risks associated with GMMs, the inactivation of their key genes responsible for stress-stimulated increase of viability and evolvability is proposed.

Acta Crystallogr D Biol Crystallogr, 2002 Feb, 58(Pt 2), 343 - 5 Epub 2002 Jan 24.
Crystallization and preliminary X-ray analysis of the molybdenum-dependent pyrogallol-phloroglucinol transhydroxylase of Pelobacter acidigallici; Abt DJ et al.; Crystals of the molybdo-/iron-sulfur protein pyrogallol:phloroglucinol hydroxyltransferase (transhydroxylase; EC 1.97.1.2) from Pelobacter acidigallici were grown by vapour diffusion in an N(2)/H(2) atmosphere using polyethylene glycol as a precipitant . In this microorganism, transhydroxylase converts pyrogallol to phloroglucinol in a unique reaction without oxygen transfer from water . Growth of crystals suitable for X-ray analysis was strongly dependent on the presence of dithionite as a reducing agent . The crystals belonged to space group P1 and MAD data were collected on the iron K edge to resolutions higher than 2.5 A.

J Bacteriol, 2002 Feb, 184(4), 1041 - 5
Speed versus efficiency in microbial growth and the role of parallel pathways; Helling RB; Many microorganisms have sets of parallel pathways for ATP production in respiration and for ATP utilization in glutamate synthesis . The alternatives differ in efficiency of ATP production and utilization . The choice among these parallel pathways has been hypothesized to control the speed and efficiency of growth . Thus, the organism should be able to alleviate (or exaggerate) deficiency in one pathway by deleting another . I show here that in Escherichia coli the effect of lack of the glutamate-synthesizing enzyme glutamate dehydrogenase on glucose-limited growth is altered predictably by ndh, cyo, and cyd mutations affecting parallel pathways leading to ATP synthesis in respiration.

N Z Med J, 2001 Nov 23, 114(1144), 519 - 21
Outbreak of cryptosporidiosis linked with a farm event; Stefanogiannis N et al.; AIM: To investigate an outbreak of cryptosporidiosis linked to a two-day farm educational event in the Wellington region . Methods . The outbreak was investigated by carrying out a site visit and interviewing cases (or their parents) identified through notifications to Wellington Regional Public Health . RESULTS: Twenty confirmed cases were linked to the event . Nineteen were aged under seven years . The most likely route of infection was the hand-to-mouth transfer of the parasite after touching an infected animal . CONCLUSIONS: To our knowledge this is the first outbreak of cryptosporidiosis associated with a farm event reported in New Zealand . Farm animals, particularly calves, are likely to carry cryptosporidiosis and other microorganisms that are pathogenic in humans . This outbreak highlights the need for organisers of similar events to implement preventive measures such as hand washing facilities and reminders . Adults supervising young children should also be vigilant in enforcing hand washing following animal contact.

Adv Nutr Res, 2001, 10, 39 - 65
Immunological activities associated with milk; Kelleher SL et al.; Milk contains a multitude of components that can, or may, provide immune protection to the suckling offspring and that also may promote development of neonatal immune competence . In addition, these specialized factors are essential for the protection of the mammary gland, the offspring's food source, from pathogen colonization and lactation failure . Breast milk also facilitates the establishment of a gut flora that inhibits colonization by many pathogens and stimulates the growth of beneficial microorganisms . Maternal immunity can be transferred to the infant via antibodies, primarily of the sIgA type in humans, as well as by leukocytes including effector and memory T lymphocytes . In this way, protection is provided passively against the pathogens to which the mother has been exposed . Currently, there is much interest in determining the protective efficacy of oral supplementation with immunoglobulins from the milk of lactating animals hyperimmunized against specific pathogens . An array of immunostimulatory components in milk, notably cytokines, may be protected against intestinal proteolysis, thereby providing the offspring with a prepackaged immune response system . These components may help to boost the infant's immature immune system . At the same time, anti-inflammatory factors in breast milk help to modulate cytokine responses to infection, thereby facilitating defense while minimizing tissue damage such as that which occurs in infants with necrotizing enterocolitis . Undoubtedly, the many components constituting the repertoire of immune and immunomodulating agents in milk interact synergistically to protect both the mammary gland and the offspring from invading pathogenic microorganisms.

Crit Care Med, 2001 Dec, 29(12), 2310 - 7
Apoptosis in cells of bronchoalveolar lavage: a cellular reaction in patients who die with sepsis and respiratory failure; Liacos C et al.; OBJECTIVE: Apoptosis represents a physiologic clearance mechanism in human tissues . The role of apoptosis has not been examined in lung cell populations, such as alveolar macrophages of septic patients, an organ frequently insulted in these patients . This study was designed to examine the effect of sepsis on the apoptosis of alveolar macrophages . DESIGN: Prospective study . SETTING: Intensive care unit and surgical intensive care and trauma unit of a large university hospital in Athens, Greece . PATIENTS: Bronchoalveolar lavage was obtained from 20 consecutive patients who met the criteria for sepsis, admitted to two intensive care units . Bronchoalveolar lavage was obtained from nine volunteers without lung disease who served as controls . INTERVENTIONS: None . MEASUREMENTS AND MAIN RESULTS: The specimens were analyzed by using annexin V binding, terminal deoxynucleotidyl transfer-mediated deoxyuridine 5-triphosphate nick end labeling (TUNEL), DNA laddering, light microscopy, and immunohistochemistry . Spontaneous apoptosis of bronchoalveolar lavage cells and particularly of alveolar macrophages was significantly decreased in septic patients compared with nonseptic controls . This finding was confirmed by using morphologic criteria and the TUNEL method . Furthermore, gel electrophoresis of DNA obtained from bronchoalveolar cells revealed that DNA fragmentation was not necessarily associated with apoptotic cell death . The bcl-2 gene was minimally expressed in the control group . An inverse correlation was found between the percentage of apoptotic alveolar macrophages and the severity of sepsis . CONCLUSIONS: The prolonged survival of lung cells in septic patients and especially of alveolar macrophages may be attributable to the inhibition of apoptosis . This seems to represent an initial attempt of the host to increase the defense capacity to kill the invading microorganism, resulting in an unbalanced tissue load of cells and an uncontrolled release of toxic metabolites . Furthermore, the inhibition of apoptosis in septic patients may explain why lung function is impaired, leading to sepsis-induced acute respiratory distress syndrome and death.

Crit Care Med, 2001 Dec, 29(12), 2303 - 9
Mortality rate attributable to ventilator-associated nosocomial pneumonia in an adult intensive care unit: a prospective case-control study; Bercault N et al.; OBJECTIVE: To evaluate the mortality rate attributable to nosocomial ventilator-associated pneumonia in an intensive care unit . DESIGN: Prospective, matched, risk-adjusted cohort study . SETTING: A 18-bed adult medical-surgical intensive care unit in a 1,100-bed regional and teaching hospital in France . PATIENTS: From January 1, 1996, to April 30, 1999, 135 patients who developed nosocomial pneumonia were matched with 135 control patients without nosocomial pneumonia . INTERVENTIONS: None . MEASUREMENTS AND MAIN RESULTS: Nosocomial pneumonia was identified on the basis of results of distal bronchial samples . The matching process was conducted according to the following primary criteria: cause of admission, indication for ventilatory support, immunologic status, cardiac status, probability of death (+/-5%), Glasgow Coma Scale score (+/-2 points), age (+/-7 yrs), and duration of exposure to risk . When possible, case and control patients were matched according to five secondary criteria: respiratory and alcoholism status before admission, diagnosis categories, surgical procedure or not, and gender . The mortality rates were compared between case and control patients by using the Kaplan-Meier estimate and the log-rank test . The influence of nosocomial pneumonia on mortality rate then was tested by adjusting for the secondary criteria and other possible confounding factors by using the Cox proportional-hazards model . The matching process was successful for 1,080 of 1,080 primary criteria . The crude intensive care unit mortality rate was higher in patients with nosocomial pneumonia than in control patients (41 vs . 14%; p <.0001) . In actuarial survival analysis, the probability of intensive care unit death was higher in the case patients (odds ratio = 2.7, 95% confidence interval = 1.8-3.1, p =.028) . After adjustment, the occurrence of nosocomial pneumonia remained an independent risk factor of death (odds ratio = 2.1, 95% confidence interval = 1.2-3.6, p =.008) . Nosocomial pneumonia attributable to multiresistant microorganisms was significantly associated with death (odds ratio = 2.6, 95% confidence interval = 1.1-5.8, p =.02) . The length of intensive care unit stay was higher in case than in control patients (31 +/- 19 vs . 26 +/- 17 days, p <.0001) . CONCLUSIONS: Nosocomial pneumonia is independently associated with death in the intensive care unit . In addition, it increases the length of intensive care unit stay.

Cell Growth Differ, 2002 Jan, 13(1), 27 - 38
Expression of toll-like receptors 2 and 4 and CD14 during differentiation of HL-60 cells induced by phorbol 12-myristate 13-acetate and 1 alpha, 25-dihydroxy-vitamin D(3); Li C et al.; Macrophages form a crucial bridge between the innate and adaptive immune response . One of their most important functions is to recognize infectious microorganisms . Toll-like receptors (TLRs) are key elements in pathogen recognition, and among them, TLR2 and TLR4 are most discussed . However, expression patterns of TLRs during myeloid cell differentiation to macrophage are unknown . In this study, we examined differentiation in the model human myeloid cell line, HL-60, treated with phorbol 12-myristate 13-acetate (PMA) or VitD(3) . Expression of TLR2, TLR4, and CD14 were measured by reverse transcription-PCR, RNase protection assay, and fluorescence-activated cell sorter assays . After treatment by PMA (1, 10, and 100 nM) for 12, 24, and 48 h, expression of TLR2 and CD14 mRNA was increased in a time- and dose-dependent manner . However, VitD(3) only induced expression of CD14 but not TLR2 in HL-60 cells . TLR4 was expressed constitutively before differentiation and increased slightly after that . Thus, PMA-mediated differentiation of HL-60 cells to macrophages is associated largely with TLR2 expression and, to a much lesser extent, with TLR4 . Furthermore, up-regulation of TLR2 and CD14 mRNA expression by PMA was abrogated by a protein kinase C inhibitor, Calphostine C, suggesting the up-regulation of TLR2 and CD14 mRNA is dependent on the activation of protein kinase C . Coexpression of CD14/TLR2 and/or CD14/TLR4 may be essential but not sufficient for the production of tumor necrosis factor-alpha in response to lipopolysaccharide in our system.

Nature, 2002 Jan 17, 415(6869), 312 - 5
A hydrogen-based subsurface microbial community dominated by methanogens; Chapelle FH et al.; The search for extraterrestrial life may be facilitated if ecosystems can be found on Earth that exist under conditions analogous to those present on other planets or moons . It has been proposed, on the basis of geochemical and thermodynamic considerations, that geologically derived hydrogen might support subsurface microbial communities on Mars and Europa in which methanogens form the base of the ecosystem . Here we describe a unique subsurface microbial community in which hydrogen-consuming, methane-producing Archaea far outnumber the Bacteria . More than 90% of the 16S ribosomal DNA sequences recovered from hydrothermal waters circulating through deeply buried igneous rocks in Idaho are related to hydrogen-using methanogenic microorganisms . Geochemical characterization indicates that geothermal hydrogen, not organic carbon, is the primary energy source for this methanogen-dominated microbial community . These results demonstrate that hydrogen-based methanogenic communities do occur in Earth's subsurface, providing an analogue for possible subsurface microbial ecosystems on other planets.

Antimicrob Agents Chemother, 2002 Feb, 46(2), 409 - 12
Effect of prolonged treatment with azithromycin, clarithromycin, or levofloxacin on Chlamydia pneumoniae in a continuous-infection Model; Kutlin A et al.; Persistent infections with Chlamydia pneumoniae have been implicated in the development of chronic diseases, such as atherosclerosis and asthma . Although azithromycin, clarithromycin, and levofloxacin are frequently used for the treatment of respiratory C . pneumoniae infections, little is known about the dose and duration of therapy needed to treat a putative chronic C . pneumoniae infection . In this study, we investigated the effect of prolonged treatment with azithromycin, clarithromycin, or levofloxacin on the viability of C . pneumoniae and cytokine production in an in vitro model of continuous infection . We found that a 30-day treatment with azithromycin, clarithromycin, and levofloxacin at concentrations comparable to those achieved in the pulmonary epithelial lining fluid reduced but did not eliminate C . pneumoniae in continuously infected HEp-2 cells . All three antibiotics decreased levels of interleukin-6 (IL-6) and IL-8 in HEp-2 cells, but this effect appeared to be secondary to the antichlamydial activity, as the cytokine levels correlated with the concentrations of microorganisms . The levels of IL-1beta, IL-4, IL-10, tumor necrosis factor alpha, and gamma interferon were too low to assess the effect of antibiotics . These data suggest that the dosage and duration of antibiotic therapy currently being used may not be sufficient to eradicate a putative chronic C . pneumoniae infection.

Klin Khir, 2001 Aug, (8), 13 - 4
{Complex treatment of patients with purulent-necrotic complications of diabetes mellitus}; Dmytriiev BI et al.; Biochemical, immunological investigations were performed in dynamics for identification of pathogenic microorganisms in the wound in 375 patients with complicated diabetes mellitus . Content of the APUD-system hormones was determined using radioimmune method . Conservative treatment included administration of insulinum pro injectionibus, plasm, albumin, rheopolyglycine, vitamins, antihypoxants, antibiotics, metronidazole, immunotherapy, dalargin, sandostatinum, D-phenylalanine . In preoperative and postoperative periods were widely applied measures of organism detoxication, physiotherapeutic methods . Original method of xenoskin was applied for closure of the wound and ulcerative defect . Application of proposed method of treatment had permitted to improve its result.

J Am Soc Nephrol, 2002 Jan, 13 Suppl 1, S78 - 83
On-line preparation of solutions for dialysis: practical aspects versus safety and regulations; Ledebo I; On-line preparation, i.e., continuous mixing and immediate use, was introduced for dialysis fluid in 1964, and it contributed significantly to the expansion of dialysis therapy through simplified handling, improved microbiology, and enhanced efficiency . On-line prepared replacement solution for hemofiltration was shown to be clinically safe as early as 1978, but the implementation was delayed for 20 yr because of regulatory conservatism . On-line preparation of sterile and pyrogen-free solutions for infusion is based on the use of water and concentrates that contribute a minimum of microorganisms and are mixed and distributed in a hygienically designed and maintained flow path . Ultrafilters with known retention capacity are placed in strategic positions and dimensioned to remove bacteria and endotoxins, which gives a sterility assurance level of at least six magnitudes, as required by the Pharmacopoeia for sterile products . Microbiologic testing of the fluid should be applied when designing, validating, and troubleshooting on-line systems but not for routine quality control, because it only gives retrospective information . Quality assurance has to be built into a system and the way it is operated . On-line fluid preparation, when properly performed, is safe, simple, and cost-effective and enhances the efficiency as well as the biocompatibility of dialysis therapy.

J Biotechnol, 2002 Mar 14, 94(1), 65 - 72
Overview of screening for new microbial catalysts and their uses in organic synthesis--selection and optimization of biocatalysts; Asano Y; As a typical example of screening for a microbial biocatalyst from nature, isolation of aldoxime-degrading microorganisms, characterization of a new enzyme phenylacetaldoxime dehydratase, and application of this enzyme to nitrile synthesis are described . The pathway in which aldoximes are successively degraded via nitrile in microorganisms could be named as 'aldoxime-nitrile pathway' . As an example of a post-screening procedure, a directed molecular evolution technique was successfully used to change the properties of nucleoside pyrophosphate phosphotransferase to make it suitable for synthesis of inosine-5'-monophosphate (5'-IMP) . With the mutant enzyme, the efficiency of the production of 5'-IMP, a food additive, was much improved.

J Biotechnol, 2002 Mar 14, 94(1), 3 - 13
Large scale analysis of sequences from Neurospora crassa; Schulte U et al.; After 50 years of analysing Neurospora crassa genes one by one large scale sequence analysis has increased the number of accessible genes tremendously in the last few years . Being the only filamentous fungus for which a comprehensive genomic sequence database is publicly accessible N . crassa serves as the model for this important group of microorganisms . The MIPS N . crassa database currently holds more than 16 Mb of non-redundant data of the chromosomes II and V analysed by the German Neurospora Genome Project . This represents more than one-third of the genome . Open reading frames (ORFs) have been extracted from the sequence and the deduced proteins have been annotated extensively . They are classified according to matches in sequence databases and attributed to functional categories according to their relatives . While 41% of analysed proteins are related to known proteins, 30% are hypothetical proteins with no match to a database entry . The entire genome is expected to comprise some 13000 protein coding genes, more than twice as many as found in yeasts, and reflects the high potential of filamentous fungi to cope with various environmental conditions.

Immunopharmacol Immunotoxicol, 2001 Nov, 23(4), 565 - 72
Neutrophil oxidative metabolism in aged humans: a perspective; Tortorella C et al.; During the last few years, several studies have pointed out the imbalance of immune responses with advancing age, this accounting for the increased susceptibility of elderly individuals to life-threatening diseases . This review is focussed on the role of neutrophil respiratory burst in the age-associated decline of cytotoxicity towards invading microorganisms . Particular emphysys is given to extracellular matrix proteins, acting as physiologic regulators of oxidative activity, as well as to neutrophil cytoskeleton, whose dysfunction is likely to be involved in the agonist-related defect of the coupling between beta2-dependent adhesive and oxidative events occurring in the aging . The role of oxidative metabolism in the increased incidence of apoptotic phenomena observed in neutrophils of aged following cell stimulation is also discussed.

J Cardiovasc Pharmacol, 2002 Feb, 39(2), 262 - 70
Anti-platelet and anti-thrombotic effects of triacetylshikimic acid in rats; Huang F et al.; Because shikimic acid is the key intermediate in the shikimate pathway in plants and microorganisms, shikimic acid and its derivatives have been described as herbicides and anti-microbial agents . Triacetylshikimic acid (TSA) is an acetylate derivative of shikimic acid . The possible anti-platelet activity and anti-thrombotic efficacy of TSA were evaluated and its effect on arachidonic acid (AA) metabolism and second messengers including cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) was evaluated . After oral pretreatment with TSA, adenosine diphosphate (ADP)-, collagen-, and AA-induced rat platelet aggregation was inhibited ex vivo in a dose-dependent manner . In an arteriovenous-shunt thrombosis model, oral administration of TSA resulted in a dose-dependent inhibition of thrombus growth . TSA markedly increased the cAMP level and showed no effect on the cGMP level in rat platelets . Also, no significant changes in ADP-induced thromboxane B2 formation in rat platelets or 6-keto-prostaglandin F 1alpha production from the abdominal aorta were observed after oral administration of low and medium doses of TSA (12.5 and 50 mg/kg) . Additionally, prothrombin time, activated partial thromboplastin time, and thrombin time were unchanged at effective anti-platelet doses of TSA . These results demonstrate that TSA exerts oral anti-platelet and anti-thrombotic efficacy without perturbation of systemic hemostasis in rats, which was partially concerned with the elevation of cAMP in platelets.

J Environ Qual, 2001 Nov-Dec, 30(6), 2081 - 90
Plant contamination by organic pollutants in phytoremediation; Sung K et al.; Phytoremediation is a remediation technique that involves plant uptake, transformation, accumulation, and/or volatilization of soil- and aqueous-phase contaminants or by the stimulation of microbial cometabolic activity in the rhizosphere of the plant . Even when the principal mechanism is by stimulation of bacteria, any resultant plant contamination cannot be overlooked . For the purpose of modeling, a two-compartment plant model has been developed . The model divides the plant into the shoot compartment (which can be harvested) and the root compartment (into which contaminants can accumulate) . Numerical experiments were conducted to investigate model behavior and to determine important parameters affecting plant contamination . Johnsongrass {Sorghum halepense (L.) Pers.} was used to evaluate the model behavior . The contaminants TNT (2,4,6,-trinitrotoluene) and chrysene were selected on the basis of their contrasting aqueous-phase solubilities . The results indicate that plant contamination and soil remediation by plants depend on soil properties such as soil organic carbon content, the physicochemical properties of the contaminants such as the octanol-water partition coefficient, and plant properties . The most important factor affecting plant contamination is bioavailability . As bioavailability increased, the concentrations in root and shoot compartments were predicted to increase . Microbial activities and plant contamination are closely related, which suggests that plants and microorganisms can have complementary roles in phytoremediation.

Bioelectrochemistry, 2002 Jan, 55(1-2), 107 - 12
Recent biotechnological developments of electropulsation . A prospective review; Teissie J et al.; During the last 25 years, basic research has improved our knowledge on the molecular mechanisms triggered at the membrane level by electric pulses . Applied aspects may now be used under safe conditions . Electropulsation is known as a very efficient tool for obtaining gene transfer in many species to produce genetically modified organisms (GMO) . This is routinely used for industrial purposes to transfer exogenous activities in bacteria, yeasts and plants . The method is simple and of a low cost . But electropulsation is not limited to this application for biotechnological purposes . It is known that the field-associated membrane alterations can be irreversible . The pulsed species cannot recover after the treatment . Their viability is strongly affected . This appears as a very promising technology for the eradication of pathogenic microorganisms . Recent developments are proposed for sterilization purposes . New flow technologies of field generation allow the treatment of large volumes of solution . When high flow rates are used, microorganisms are submitted both to a hydromechanical and to an electrical stress . The synergy of the two effects may be present when suitable pulsing conditions are chosen . Several examples for the treatment of domestic water and in the food industry are described . Walled microorganisms are affected not only at the membrane level . We observed that alterations are present on the cell wall . A very promising technology is the associated controlled leakage of the cytoplasmic soluble proteins . Large dimeric proteins such as beta-galactosidases can be extracted at a high yield . High volumes can be treated by using a flow process . Extraction of proteins is obtained with many systems including mammalian cells.

Mikrobiol Z, 2001 Sep-Oct, 63(5), 80 - 101
{Biological functions of microbial exopolysaccharides}; Pyrog TP; The review includes literature data and the author's experimental data concerning the role of microbial exopolysaccharides (EPS) in protection of the producer cells from different unfavourable factors (drying, effect of toxic metals, antibiotics, biocydes, etc), participation of EPS in trophic metabolism of microorganisms (use of EPS as a source of carbon supply, role in regulation of cell ion balance), as well as in interaction of producers with other microorganisms, macroorganisms, objects of inorganic nature (EPS as virulence factor, EPS participation in the processes of adhesion, sorption and aggregation, in formation of plant-bacterial symbioses) . A question of interrelation of physico-chemical properties of microbial EPS and their biological functions as well as of these data use in biotechnology for production of polysaccharides with preset properties is discussed.

Mikrobiologiia, 2001 Nov-Dec, 70(6), 796 - 803
{Stability of recombinant plasmids in transgenic microorganism cells under various environmental conditions}; Popova LIu et al.; The copy number of R plasmids weakly depends on the selective pressure of the respective antibiotic but does depend on the physiology of the host species and the type of plasmids and cloned genes, whose expression leads to a further load on the biosynthetic apparatus of cells . The last factor is critical in the maintenance of recombinant plasmids in transgenic microorganisms.

Exp Mol Pathol, 2002 Feb, 72(1), 68 - 76
Mice genetically lacking endothelial selectins are resistant to the lethality in septic peritonitis; Matsukawa A et al.; Leukocyte interactions with vascular endothelium are an initial step for leukocyte entry into infectious foci where endothelial selectins may play a key role . Infiltrating leukocyte is essential for bacterial clearance, suggesting that endothelial selectins would be important in host defense against microorganisms . To address this, E-, P-, and E/P-selectin-deficient mice (E(-/-), P(-/-), E/P(-/-)) and wild-type (WT) mice underwent cecal ligation and puncture (CLP) . Neither leukocyte infiltration nor bacterial load in the peritoneum was altered in E(-/-), P(-/-), and E/P(-/-) mice compared to WT mice . However, E(-/-), P(-/-), and E/P(-/-) mice were resistant to the lethality induced by CLP . At the mechanistic level, E(-/-), P(-/-), and E/P(-/-) mice did not develop renal dysfunction, a possible cause of death during sepsis . The serum level of interleukin-13 in E(-/-), P(-/-), and E/P(-/-) mice that had undergone CLP was higher than that in WT mice, whereas levels of macrophage inflammatory protein-2, KC in serum, and KC in kidney were lower than those in WT mice . These experiments demonstrate that endothelial selectin-mediated leukocyte rolling is not required for leukocyte entry in septic peritonitis and that endothelial selectins may affect mice survival during sepsis by influencing the cytokine profiles .

Appl Biochem Biotechnol, 2001 Oct-Dec, 96(1-3), 225 - 38
Immobilization and stabilization of biomaterials for biosensor applications; D'Souza SF; Biosensors are finding applications in a variety of analytical fields . A biosensor basically consists of a transducer in conjunction with a biologically active molecule that converts a biochemical signal into a quantifiable electric response . The specificity of the biosensor depends on the selection of the biomaterial . Enzymes, antibodies, DNA, receptors, organelles, microorganisms as well as animal and plant cells or tissues have been used as biologic sensing materials . Advances in biochemistry, molecular biology, and immunochemistry are expected to lead to a rapid expansion in the range of biologic recognition elements to be used in the field of biosensors . Biomaterials that are stable and function even in highly acidic, alkaline, hydrophobic, or oxidizing environments as well as stable to high temperature and immune to toxic substrates in the processing stream will play an important role . Techniques for immobilization of the biomaterials have played a significant role in the biosensor field . Immobilization not only brings about the intimate contact of the biologic catalysts with the transducer, but also helps in the stabilization of the biologic system, thus enhancing its operational and storage stability . A number of techniques have been developed in our laboratory for the immobilization of enzymes, multienzyme systems, cells, and enzyme-cell conjugates . Some of these aspects that are of significance in biosensor applications have been highlighted.

Environ Sci Technol, 2001 Sep 15, 35(18), 3780 - 5
Dense medium plasma environments: a new approach for the disinfection of water; Manolache S et al.; The levels to which microbial colony forming units are permitted in various waters fit for human contact are carefully regulated . Conventional chemical and physical approaches usually are complex processes with significant limitations due to the generation of toxic side-products . In this contribution a novel plasma reactor--dense medium plasma reactor--is described, and its efficiency for the disinfection of contaminated water is discussed . It has been shown that owing to the intense stirring of the reaction medium (e.g . contaminated water), as a result of the specially designed spinning electrode and gas-flow system, a volume-character discharge is created, which can efficiently kill bacteria . It has been demonstrated that treatment times as low as 20 s are enough for the total inactivation of microorganisms for 200 mL of 10(5) bacteria/mL contaminated water.

Crit Care Med, 2002 Jan, 30(1 Suppl), S1 - 11
Toll-like receptors; Lien E et al.; The ability of a host to sense invasion by pathogenic organisms and to respond appropriately to control infection is paramount to survival . In the case of sepsis and septic shock, however, an exaggerated systemic response may, in fact, contribute to the morbidity and mortality associated with overwhelming infections . The innate immune system has evolved as the first line of defense against invading microorganisms . The Toll-like receptors (TLRs) are a part of this innate immune defense, recognizing conserved patterns on microorganisms . These TLRs and their signaling pathways are represented in such diverse creatures as mammals, fruit flies, and plants . Ten members of the TLR family have been identified in humans, and several of them appear to recognize specific microbial products, including lipopolysaccharide, bacterial lipoproteins, peptidoglycan, and bacterial DNA . Signals initiated by the interaction of TLRs with specific microbial patterns direct the subsequent inflammatory response . Thus, TLR signaling represents a key component of the innate immune response to microbial infection.

Clin Microbiol Rev, 2002 Jan, 15(1), 111 - 24
Bacterial endophthalmitis: epidemiology, therapeutics, and bacterium-host interactions; Callegan MC et al.; Endophthalmitis is a severe inflammation of the interior of the eye caused by the introduction of contaminating microorganisms following trauma, surgery, or hematogenous spread from a distant infection site . Despite appropriate therapeutic intervention, bacterial endophthalmitis frequently results in visual loss, if not loss of the eye itself . Although the pathogenicity of bacterial endophthalmitis has historically been linked with toxin production during infection, a paucity of information exists as to the exact mechanisms of retinal toxicity and the triggers for induction of the intraocular immune response . Recently, research has begun to examine the bacterial and host molecular and cellular events that contribute to ocular damage during endophthalmitis . This review focuses on the causative agents and therapeutic challenges of bacterial endophthalmitis and provides current data from the analysis of the role of bacterial virulence factors and host inflammatory interactions in the pathogenesis of eye infections . Based on these and related studies, a hypothetical model for the molecular pathogenesis of bacterial endophthalmitis is proposed . Identifying and understanding the basic mechanisms of these bacterium-host interactions will provide the foundation for which novel, information-based therapeutic agents are developed in order to prevent vision loss during endophthalmitis.

Phys Med, 2001, 17 Suppl 1, 267 - 8
A device to study the effect of space radiation on photosynthetic organisms; Angelini G et al.; This research concerns the study of the effects of ionising space radiation on the oxygen-evolving activity of algae and cyanobacteria, focusing our attention on Photosystem II (PS-II), the oxygen-evolving complex . These microorganisms as higher plants, can use light energy to split water molecules and evolve oxygen in a process that produce storable energy-rich products from atmospheric carbon dioxide . Algae and cyanobacteria which can grow in the presence of nutrients and carbonate are expected to be utilised to maintain an oxygen-atmosphere and to constitute biomass in space shuttles . Irradiation was performed in gamma 60Co-sources of different activities; fluorescence techniques in vivo and SDS-PAGE analysis in vitro were used to determine PS-II efficiency during radiation stress . We determined the radiation target on PS-II by immunoblot . We built a miniaturised growth box that preserves constant pressure and temperature to measure automatically photosynthetic activity by a fluorescence sensor, directly in space during a mission in an ASI balloon.

Infect Dis Clin North Am, 2001 Dec, 15(4), 1047 - 71
Laboratory diagnosis of central nervous system infections; Thomson RB Jr et al.; The laboratory diagnosis of CNS infection is essential for optimal therapy . Acute infection requires rapid turn-around testing with high predictive values, that is, the ability of a test to accurately identify those patients who do or do not have disease caused by a specific etiology . The Gram's stain, fungal stains of direct smears, antigen testing for C . neoformans, and culture of bacteria, fungi, mycobacteria, and some viruses are important tests for the diagnosis of acute infection . The laboratory diagnosis of chronic infection necessitates discussion between the clinician and laboratory technician to allow triaging of testing . Antigen tests for bacteria, fungi, and viruses; antibody tests for multiple microorganisms; and PCR testing for bacteria, M . tuberculosis, and many viruses are all important in limited clinical situations . All testing for acute or chronic disease depends on sufficient specimen that is transported to the laboratory in a manner that will not compromise viability or chemical integrity . Sterile containers that maintain moisture content, exclude oxygen for anaerobic requests, and are stored at proper temperatures (22 degrees C room, 4 degrees C refrigeration, or -20 degrees C freezer depending on pathogen and test) are mandatory . Many laboratory issues addressing the diagnosis of CNS infection are changing or evolving . Most important is the recognition that bacterial antigen testing for the diagnosis of acute bacterial meningitis rarely impacts patient management and is not routinely needed, CSF shunt infections differ from usual meningeal infections and require rapid diagnosis, and TB meningitis remains a difficult disease to diagnosis but may be confirmed first by PCR testing of CSF . In addition, Whipple's disease of the CNS can be confirmed using PCR with CSF; CJD has a marker protein, referred to as 14-3-3 antigen, that can be detected in CSF, and the diagnosis of fungal CNS disease requires careful interpretation of direct smears, antigen and antibody testing, and culture . Most difficult to diagnose among the CNS infections are viral meningitis and encephalitis . The appearance of new etiologies, such as West Nile virus, and the common use of PCR for the herpes viruses and enteroviruses represent important advances . Evolving methods for the laboratory diagnosis of CNS infection represent significant improvements over previous testing; however, the array of tests available demands more attention for appropriate selection, is significantly more expensive, and requires new skills for performance and interpretation . The responsibility for proper use of laboratory testing lies both with the clinician and laboratory technician.

Biochem Biophys Res Commun, 2002 Jan 11, 290(1), 339 - 48
Salt stress and hyperosmotic stress regulate the expression of different sets of genes in Synechocystis sp . PCC 6803; Kanesaki Y et al.; Acclimation of microorganisms to environmental stress is closely related to the expression of various genes . We report here that salt stress and hyperosmotic stress have different effects on the cytoplasmic volume and gene expression in Synechocystis sp . PCC 6803 . DNA microarray analysis indicated that salt stress strongly induced the genes for some ribosomal proteins . Hyperosmotic stress strongly induced the genes for 3-ketoacyl-acyl carrier protein reductase and rare lipoprotein A . Genes whose expression was induced both by salt stress and by hyperosmotic stress included those for heat-shock proteins and the enzymes for the synthesis of glucosylglycerol . We also found that each kind of stress induced a number of genes for proteins of unknown function . Our findings suggest that Synechocystis recognizes salt stress and hyperosmotic stress as different stimuli, although mechanisms common to the responses to each form of stress might also contribute to gene expression . (c)2002 Elsevier Science.

Blood Cells Mol Dis, 2001 Jul-Aug, 27(4), 728 - 30
Synergy between TLR2 and TLR4: a safety mechanism; Beutler E et al.; BACKGROUND: The Toll (Tlr) receptors facilitate innate immunity by detecting products that are unique to invading microorganisms . Stimulation of these receptors can produce severe reactions and death . We propose that synergy between receptors for different microbial products would provide a safety mechanism, preventing inappropriate, potentially fatal reactions by reacting to low concentrations of ligands when more than a single ligand is present . RESULTS: Striking synergy is noted between the ligand for Tlr4, lipopolysaccharide (LPS), and a ligand for Tlr2, muramyl dipeptide (MDP), in the release of tumor necrosis factor from RAW cells . CONCLUSIONS: Synergy between a ligand for Tlr2 and Tlr4 can be demonstrated in a simple in vitro system . The greater sensitivity of MDP-stimulated cells to LPS may explain the data that were once interpreted incorrectly as indicating that Tlr2 is the endotoxin receptor.

Support Care Cancer, 2002 Jan, 10(1), 58 - 64
Fever and neutropenia in children with solid tumors is similar in severity and outcome to that in children with leukemia; Kocak U et al.; There is growing interest in developing criteria that will allow efficient prospective discrimination between cancer patients at high and at low risk for complex fever and neutropenia . The objective of this study was to determine whether there were differences in patterns of documented infections and outcome of episodes of fever and neutropenia in pediatric patients with leukemia and those with solid tumors, a potential risk factor . A total of 283 febrile neutropenia episodes in pediatric cancer patients at a single center were retrospectively reviewed; 38% of the patients concerned had leukemia and 62% had solid tumors . Fever of unexplained origin was seen in 73% and 74% of episodes in patients with leukemia and solid tumor, respectively . Bacteremia occurred in 18% and 16% of patients in these respective groups . There was no difference in the type of microorganisms that were isolated in the groups, with gram positives predominating in both . The median duration of fever was 2 days in both groups . The depth of neutropenia was similar, with 75% of leukemia patients and 70% of solid tumor patients presenting with ANC of 100 cells/microl or lower . The median duration of neutropenia was 9 days in patients with leukemia and 6 days in solid tumor patients . The median duration of antibiotic treatment was 9 days and 7.5 days in the same respective groups . Antibiotic modification occurred in 25% episodes of febrile neutropenia in leukemia patients and in 11% of episodes in solid tumor patients . No deaths occurred in either group . Subgroup analysis of leukemic patients suggested that patients in the induction phase of therapy have a higher rate of bacteremia and pneumonia . No substantial difference in course or outcome was seen between the leukemia and solid tumor groups, possibly because of the intensive treatment administered to pediatric patients with solid tumors . Risk assessment strategies based on chemotherapy dose intensity and patient comorbidities rather than underlying malignancy should be prospectively studied.

J Microbiol Methods, 2002 Feb, 48(2-3), 207 - 19
Culture of the marine cyanobacterium, Lyngbya majuscula (Oscillatoriaceae), for bioprocess intensified production of cyclic and linear lipopeptides; Burja AM et al.; Cyanobacteria are an ancient and diverse group of photosynthetic microorganisms, which inhabit many different and extreme environments . This indicates a high degree of biological adaptation, which has enabled these organisms to thrive and compete effectively in nature . The filamentous cyanobacterium, Lyngbya majuscula, produces several promising antifungal and cytotoxic agents, including laxaphycin A and B and curacin A . Samples of L . majuscula collected from Moorea Island, Tahiti (French Polynesia) and from the Culture Collection of Algae and Protozoa (CCAP 1446/4) were studied and adapted to large scale laboratory culture (5 l) . This constitutes a 100-fold scale-up for the culture of this particular strain of L . majuscula . The effect of culture vessel configurations, growth conditions and media compositions on growth of L . majuscula was examined . Using optimised culture conditions, two strains of L . majuscula are currently being evaluated for their production of secondary metabolites . Results will be compared with those obtained from four environmental extracts . Comparisons were made by thin layer chromatography (TLC), high performance liquid chromatography (HPLC) and fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) . It was shown that varying the culture conditions under which L . majuscula was grown had the greatest effect on secondary metabolite production, thus providing potential for future bioprocess intensified production.

J Microbiol Methods, 2002 Feb, 48(2-3), 127 - 38
Rapid typing of bacteria using matrix-assisted laser desorption ionisation time-of-flight mass spectrometry and pattern recognition software; Bright JJ et al.; Matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) of intact microorganisms, also known as intact cell MALDI-TOF-MS (ICM-MS), has been shown to produce characteristic mass spectral fingerprints of moieties desorbed from the cell surface . ICM-MS spectra can be obtained in minutes after removal of a colony from a culture plate . The similarity of ICM-MS spectra of replicate samples and of two different batches of the same bacterial strain demonstrates, in this study, the reproducibility of the technique . We have developed the Manchester Metropolitan University Search Engine (MUSE) to rapidly build and search databases of ICM-MS spectra . A database of 35 strains, representing 20 species and 12 genera, was built with MUSE and used to identify 212 isolates . The database was created in 26 s and loaded in 10 s, ready for searching, which took less than 1 s per isolate . Correct matches were made in 79%, 84% and 89% of the 212 samples at strain, species and genus levels, respectively . At least 50% of the replicates of 42 of the 45 isolates matched the correct strain, and the most commonly identified species for 43 of the 45 isolates was the correct one . The close match of the Escherichia coli strains containing the O157 antigen and the E . coli strains containing the K1 antigen suggests that these antigens may have a dominating influence on the ICM-MS fingerprints of these strains . We now have the ability to acquire ICM-MS fingerprints of bacteria and to search a database of these fingerprints within minutes, so that the rapid identification of bacteria to the strain level can be realised.

Rev Environ Contam Toxicol, 2002, 173, 39 - 83
Toxicity of azaarenes; Bleeker EA et al.; Heterocyclic compounds by far outnumber the homocyclic PAHs . In addition, they are often more soluble in water, which may imply a greater biological significance of these heterocycles . Yet, most research focuses on the homocyclics, based on the implicit assumption that the mostly higher concentration of the homocyclics rank these compounds as priority compounds . This review critically examines the available evidence and poses questions on the biological activity and environmental risk of one small group of heterocyclics, the azaarenes, which contain one nitrogen atom in one of the aromatic rings . In different sections, the biotransformation and different types of toxicity are discussed in comparison to those of homocyclic PAHs . The last section focuses on the implications for risk assessment of PAHs . Two- and three-ringed azaarenes can be relatively easily transformed by bacteria, fungi, invertebrates, and vertebrates . The presence of the N-moiety in the smaller azaarenes leads to metabolic routes that partly differ from those of the homoaromatic analogues . Major metabolic products of the azaarenes appear to be ketones and mono- or dihydroxylated azaarenes . Microorganisms can further degrade these into multiple oxygen-containing compounds or they can open up the aza-containing aromatic ring and fully metabolize the products . Fungi and vertebrates were shown to produce the mutagenic dihydrodiol metabolites . The metabolism of the larger azaarenes in vertebrates proceeds analogous to homoaromatic PAH, because in these larger molecules the N-moiety has less influence . Transformation of the larger azaarenes by microorganisms proceeds much slower if occurring at all . Direct toxicity data of azaarenes are mostly restricted to the effects of acridine and quinoline on a relatively small number of species . From this limited set it becomes clear that differences between species are relatively small . As with homocyclic PAHs, toxicity generally increases with increasing number of rings, and baseline toxicity models based on homocyclic PAHs do apply . Toxicity differences between isomers indicate that azaarene toxicity cannot be explained by molecular size-related parameters alone, indicating that electronic forces may be important as well . Considering chronic toxicity it becomes clear that the often-used acute-to-chronic-ratios often underestimate specific chronic toxicity, even within the very limited set of chronic data available . In contrast with homocyclic PAHs, photodegradation of azaarenes shows the same degradation products as biological transformation involving monooxygenases . In general, as for homocyclic PAHs, the degree of phototoxicity is related to the UV absorption characteristics of the azaarenes, which makes it possible to apply the QSAR models developed for homocyclic PAHs to azaarenes as well . Recent research on algae showed that UV-A is the main cause of photoenhanced toxicity . Together with the fact that in the water column UV-B is almost absent, this clearly demonstrates the relevance of phototoxicity in the field . Mutagenicity of azaarenes generally proceeds through similar pathways as in homocyclic PAHs, with bay region diol epoxides as major genotoxic metabolites . The N-moiety can, however, result in differences in genotoxic activities between isomers . Carcinogenicity of azaarenes in mammals is generally restricted to four-ringed and larger structures, and mechanisms leading to cancer are similar to those of homocyclic aromatics . An exception to this general pattern is quinoline, which has been shown to induce liver cancer . The present risk assessment for PAHs is solely based on homocyclic PAHs . Yet, from the present review it becomes clear that this approach fails to protect against a vast number of heterocyclic compounds and biotransformation products that may exhibit stronger or other toxic effects than their homocyclic analogues . Therefore, incorporating the role of heterocyclic compounds and their metabolism appears to be a necessity for a reliable risk assessment for polycyclic aromatic compounds . In addition, reliable long-term protection against PAHs demands data on chronic toxicity, including teratogenicity, both for homocyclic as for heterocyclic compounds.

Rev Environ Contam Toxicol, 2002, 173, 117 - 41
RMS Titanic and the emergence of new concepts on consortial nature of microbial events; Cullimore DR et al.; The RMS Titanic sank in 1912 and created a historical event that still ripples through time . Stories were told and lessons learned but the science has only just begun . Today the fading remains of the ship resemble the hanging gardens of Babylon except that it is not plants that drape the walls but complex microbial growths called rusticles . These organisms have been found to be not a species, like plants and animals, but to be structures created by complex communities of bacterial species . Like the discovery of tube worms in the mid-oceanic vents, the nature of these rusticles presents another biological discovery of a fundamental nature . Essentially these microbial consortia on the RMS Titanic have generated structures of a mass that would rival whales and elephants while gradually extracting the iron from the steel . Rusticle-like consortia appear to play many roles within the environment, and it is perhaps the RMS Titanic that is showing that there is a new way to understand the form, function, and nature of microorganisms . This understanding would develop by considering the bacteria not as individual species functioning independently but as consortia of species functioning in community structures within a common habitat . This concept, if adopted, would change dramatically the manner in which a microbial ecologist and any scientist or engineer would view the occurrence of a slime, encrustation, biocolloid, rust flake, iron pan, salt deposit, and perhaps even some of the diseases that remain unexplained as a disease of unknown cause.

J Antibiot (Tokyo), 2001 Oct, 54(10), 805 - 9
New biotransformation products of nemadectins; Fang K et al.; Selected nemadectins (formerly LL-F28249 series) have been fed to a panel of microorganisms with the aim of generating new derivatives . In addition to products resulting from the oxidation of the terminal methyl group (C-29), a unique phosphorylated nemadectin was isolated . The phosphate group was determined to be at C-23 by HMBC between phosphorus and H-23 . Milbemycin or nemadectin derivatives with natural substituents involving the 23-hydroxyl group were hitherto unknown.

J Ind Microbiol Biotechnol, 2001 Dec, 27(6), 352 - 6
Molecular genetics and industrial microbiology--30 years of marriage; Demain AL; Thirty years ago, molecular genetics and industrial microbiology joined their hands in marriage . The event took place in Prague at the first Symposium on the Genetics of Industrial Microorganisms . My closing plenary lecture, titled "The Marriage of Genetics and Industrial Microbiology--After a long Engagement, a Bright Future," dealt with industrial uses of mutants, the lack of success with genetic recombination, control of branched and unbranched pathways and thoughts about the future, e.g., identifying the biochemical sites of beneficial mutations, exploitation of recombination and genetic means to increase production of enzymes . It is quite amazing that the Symposium was held 3 years before the advent of recombinant DNA technology . This important meeting was followed in 1976 by the first Genetics and Molecular Biology of Industrial Microorganisms (GMBIM) meeting in Orlando, all of the six subsequent GMBIM meetings being held in Bloomington, Indiana . Today, thousands of biotechnology companies are in existence making great progress in the pharmaceutical and agricultural sectors . Hundreds of new genetically engineered compounds, produced in microbial, mammalian or insect cells, are in clinical trails and many are already being marketed . The field is booming with new technologies such as transgenic animals and plants, site-directed mutagenesis, combinatorial biosynthesis, gene therapy, antisense, abzymes, high-throughput screening, monoclonal antibodies, PCR and many more . Agricultural biotechnology has made great strides but unfortunately its progress is being delayed by political controversy.

J Clin Oncol, 2002 Jan 1, 20(1), 110 - 24
Pharmacodynamic studies of the epidermal growth factor receptor inhibitor ZD1839 in skin from cancer patients: histopathologic and molecular consequences of receptor inhibition; Albanell J et al.; PURPOSE: The epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor ZD1839 (Iressa; AstraZeneca Pharmaceuticals, Alderley Park, United Kingdom) is under development as an anticancer agent . We studied the pharmacodynamic effects of ZD1839 on EGFR in the skin, an EGFR-dependent tissue, in cancer patients participating in ZD1839 phase I clinical trials . PATIENTS AND METHODS: We studied 104 pre- and/or on-ZD1839 therapy ( approximately at day 28 of therapy) skin biopsies from 65 patients receiving escalating doses of daily oral ZD1839 . We measured ZD1839 effects on EGFR activation by immunohistochemistry using an antibody specific for the activated (phosphorylated) EGFR . Effects on receptor signaling (activated mitogen-activated protein kinase {MAPK}), proliferation, p27(KIP1), and maturation were also assessed . RESULTS: Histopathologically, the stratum corneum of the epidermis was thinner during therapy (P <.001) . In hair follicles, prominent keratin plugs and microorganisms were found in dilated infundibula . ZD1839 suppressed EGFR phosphorylation in all EGFR-expressing cells (P <.001) . In addition, ZD1839 inhibited MAPK activation (P <.001) and reduced keratinocyte proliferation index (P <.001) . Concomitantly, ZD1839 increased the expression of p27(KIP1) (P <.001) and maturation markers (P <.001) and increased apoptosis (P <.001) . These effects were observed at all dose levels, before reaching dose-limiting toxicities . CONCLUSION: ZD1839 inhibits EGFR activation and affects downstream receptor-dependent processes in vivo . These effects were profound at doses well below the one producing unacceptable toxicity, a finding that strongly supports pharmacodynamic assessments to select optimal doses instead of a maximum-tolerated dose for definitive efficacy and safety trials.

Biofizika, 2001 Nov-Dec, 46(6), 1048 - 61
{The use of various models of biosynthesis kinetics}; Arzamastsev AA et al.; The available models of the microorganism growth kinetics are analyzed . The possibility of application of the Monod model is studied . It is shown that these models do not permit describing adequately the experimental data at different initial concentrations of biomass and substrate that vary in a wide range . The calculations of the static modes of operation of biochemical react ors differ greatly.

Microbiol Res, 2001, 156(4), 303 - 9
Microorganisms asssociated with Withania somnifera leaves; Alwadi HM et al.; Microorganisms including bacteria, actinomycetes and fungi were recovered from the leaves of Withania somnifera, which were collected from two altitudinal ranges (0-300 m and 1700-2000 m) in the Asir region, Saudi Arabia . Types and numbers of microorganisms varied according to the altitude and the month of collection . The number of microorganisms was higher on old leaves than that on young ones in most cases . Low altitude exhibited more microorganisms than high altitude . The relationship between meteorological factors and type and number of the recovered microorganisms is discussed . Inoculation of detached healthy leaves of Withania by all recovered fungal species revealed only Alternaria solani as a pathogen of this plant . To confirm pathogenicity, scanning and transmission electron microscopic examination revealed the colonization of this pathogen inside the leaf tissue . Penetration of Withania leaves by the fungus occurred only through stomata, and the invading hyphae were located in the intercellular spaces of leaf tissues . Ultrastructural changes noted in infected cells included changes in chloroplasts and the invagination of the host plasma membrane.

Environ Sci Technol, 2001 Dec 1, 35(23), 4592 - 6
Emission of chiral organochlorine pesticides from agricultural soils in the cornbelt region of the U.S; Leone AD et al.; Several organochlorine pesticides are chiral molecules manufactured as racemic mixtures . Past research has shown that selective degradation of pesticide enantiomers by microorganisms occurs resulting in nonracemic signatures in soils . In this work, volatilization of chiral pesticides from soil was investigated to determine if enantioselective breakdown in soils could be used as a source signature to track releases of chiral pesticides to the atmosphere . Air samples were taken directly above agricultural soils at several sites, and enantiomeric signatures were found to be nonracemic following patterns found in the soil . A follow up study at one site showed that for most compounds concentration decreased with increasing height above the soil, while enantiomer fractions for chiral pesticides were similar to that found in the soil, signifying the soil as a source to the air . The enantiomer fractions of ambient air samples from rural nonagricultural areas in the region were also found to be nonracemic.

Ann Allergy Asthma Immunol, 2001 Dec, 87(6 Suppl 3), 57 - 63
Stachybotrys: relevance to human disease; Terr AI; LEARNING OBJECTIVES: Recent public concern about the danger of environmental fungi has focused attention on one particular mold, Stachybotrys . The purpose of this review is to examine and critique the published literature on Stachybotrys for objective scientific and clinical evidence of disease caused by the presence of this fungal organism in the environment . DATA SOURCES: Data were obtained from all published research and reviews of Stachybotrys indexed in MEDLINE since 1966 . STUDY SELECTION: The publications used for this review were those that contained information about human health effects of this microorganism . The critique of these publications is the author's . RESULTS: Stachybotrys is a minor component of the indoor mycoflora, found on certain building material surfaces in water-damaged buildings, but airborne spores are present in very low concentrations . Published reports fail to establish inhalation of Stachybotrys spores as a cause of human disease even in water-damaged buildings . A possible exception may be mycotoxin-caused pulmonary hemorrhage/hemosiderosis in infants, although scientific evidence to date is suggestive but not conclusive . Based on old reports ingestion of food prepared from Stachybotrys-contaminated grains may cause a toxic gastroenteropathy . No convincing cases of human allergic disease or infection from this mold have been published . CONCLUSIONS: The current public concern for adverse health effects from inhalation of Stachybotrys spores in water-damaged buildings is not supported by published reports in the medical literature.

J Food Prot, 2001 Dec, 64(12), 2088 - 102
Confocal microscopy and microbial viability detection for food research; Takeuchi K et al.; Confocal microscopy offers several advantages over other conventional microscopic techniques as a tool for studying the interaction of bacteria with food and the role of food microstructure in product quality and safety . When using confocal microscopy, samples can be observed without extensive preparation processes, which allows for the evaluation of food without introducing artifacts . In addition, observations can be made in three dimensions without physically sectioning the specimen . The confocal microscope can be used to follow changes over a period of time, such as the development of the food structure or changes in microbial population during a process . Microbial attachment to and detachment from food and food contact surfaces with complex three-dimensional (3-D) structures can be observed in situ . The fate of microbial populations in food system depends on processing, distribution, and storage conditions as well as decontamination procedures that are applied to inactivate and remove them . The ability to determine the physiological status of microorganisms without disrupting their physical relationship with a food system can be useful for determining the means by which microorganisms survive decontamination treatments . Conventional culturing techniques can detect viable cells; however, these techniques lack the ability to locate viable cells in respect to the microscopic structures of food . Various microscopic methods take advantage of physiological changes in bacterial cells that are associated with the viability to assess the physiologic status of individual cells while retaining the ability to locate the cell within a food tissue system . This paper reviews the application of confocal microscopy in food research and direct observation of viable bacteria with emphasis on their use in food microbiology.

J Food Prot, 2001 Dec, 64(12), 2078 - 82
A new kinetic model for thermal inactivation of microorganisms: development and validation using Escherichia coli O157:H7 as a test organism; Huang L et al.; A new kinetic model has been proposed to simulate the nonlinear behavior of survivor curves frequently observed in thermal inactivation of microorganisms . This model incorporates a time component into the first-order inactivation kinetics and is capable of describing the linear, convex, and concave survivor curves . The model was validated using Escherichia coli O157:H7 as a test microorganism . Ground beef (93% lean) samples inoculated to 10(7) to 10(8) CFU/g of meat were subjected to immersion heating at 55, 57.5, 60, 62.5, and 65 degrees C, respectively, in a water bath . All the survivor curves in this study showed upward concavity . Linear and nonlinear regressions were used to fit the survivor curves to the linear first-order inactivation kinetics and the proposed model . Analyses showed that the new kinetic model provides a much better estimate of the thermal inactivation behavior of E . coli O157:H7 in ground beef.

Clin Lab Med, 2001 Dec, 21(4), 897 - 909, x-xi
Microbiology; Shapiro HM; Although flow cytometry can detect microorganisms rapidly in low concentrations in some clinical samples, it has not proved cost-effective for clinical use in this application . Adaptation of fluorescence multiplexing methodology recently developed for flow cytometric bead immunoassays, and the introduction of hybrid cytometers based on microfluidic technology, however, may make it possible to place cost-effective cytometric apparatus in clinical microbiology laboratories in the near future.

Rev Saude Publica, 2001 Jun, 35(3), 315 - 7
{Mycobacterium avium complex in water buffaloes slaughtered for consumption}; Freitas J et al.; Two mycobacterium strains isolated from lung tissue a apical lymph nodes of slaughtered water buffaloes were biochemically analyzed and identified as Mycobacterium avium complex strains . Association between these microorganisms and the acquired immunodeficiency syndrome, and the potential risk posed by eating infected animals and their products, was discussed.

Environ Toxicol, 2001, 16(6), 523 - 34
Characterisation of a gene cluster involved in bacterial degradation of the cyanobacterial toxin microcystin LR; Bourne DG et al.; A novel pathway for degradation of the cyanobacterial heptapeptide hepatotoxin microcystin LR was identified in a newly isolated Sphingomonas sp . (Bourne et al . 1996 Appl . Environ . Microbiol . 62: 4086-4094) . We now report the cloning and molecular characterisation of four genes from this Sphingomonas sp . that exist on a 5.8-kb genomic fragment and encode the three hydrolytic enzymes involved in this pathway together with a putative oligopeptide transporter . The heterologously expressed degradation pathway proteins are enzymatically active . Microcystinase (MlrA), the first enzyme in the degradative pathway, is a 336-residue endopeptidase, which displays only low sequence identity with a hypothetical protein from Methanobacterium thermoautotrophicum . Inhibition of microcystinase by EDTA and 1,10-phenanthroline suggests that it is a metalloenzyme . The most likely residues that could potentially chelate an active-site transition metal ion are in the sequence HXXHXE, which would be unique for a metalloproteinase . Situated immediately downstream of mlrA with the same direction of transcription is a gene mlrD, whose conceptual translation (MlrD, 442 residues) shows significant sequence identity and similar potential transmembrane spanning regions to the PTR2 family of oligopeptide transporters . A gene mlrB is situated downstream of the mlrA and mlrD genes, but transcribed in the opposite direction . The gene encodes the enzyme MlrB (402 residues) which cleaves linear microcystin LR to a tetrapeptide degradation product . This enzyme belongs to the "penicillin-binding enzyme" family of active site serine hydrolases . The final gene in the cluster mlrC, is located upstream of the mlrA gene and is transcribed in the opposite direction . It codes for MlrC (507 residues) which mediates further peptidolytic degradation of the tetrapeptide . This protein shows significant sequence identity to a hypothetical protein from Streptomyces coelicolor . It is suspected to be a metallopeptidase based on inhibition by metal chelators . It is postulated on the basis of comparison with other microorganisms that the genes in this cluster may all be involved in cell wall peptidoglycan cycling and subsequently act fortuitously in hydrolysis of microcystin LR.

Fresenius J Anal Chem, 2001 Nov, 371(5), 675 - 81
Analytical determination of the microbial utilization and transformation of humic acids extracted from municipal refuse; Filip Z et al.; Humic substances are usually the refractory part of natural organic matter, and in a landfill they can retain inorganic and organic micropollutants . This study has investigated analytically whether humic acids (HA) extracted by use of alkali from either fresh municipal refuse or from refuse disposed of in a landfill for up to 12 months can resist microbial degradation under aerobic conditions . When added as a supplementary nutrient source, up to 63.6% of HA was utilized and this percentage was enhanced to a mean value of 88.5% when different HA preparations were used as the sole source of carbon . In cultures of a soil microbial community containing the same preparations as sole sources of nitrogen, HA was usually completely utilized . The remaining HA re-isolated from some microbial cultures were highly depleted in carbon and, simultaneously, the nitrogen content was enhanced . The FTIR spectra were indicative of strong participation of aliphatic structural units in the refuse-related HA preparations . Because of the microbial activity, different carbonaceous substances were primarily removed from the HA structure, and an increase in nitrogenous molecular groups became apparent . The structural transformations brought about by soil microorganisms "in vitro" corresponded to those occurring naturally in HA obtained from refuse aged for 12 months in a landfill.

Crit Care Nurse, 2001 Dec, 21(6), 35 - 47
Understanding the neonatal immune system: high risk for infection; McKenney WM; Neonates are vulnerable to infection from prenatal and postnatal exposure to microorganisms . The immaturity of the immune system in neonates and illness and/or prematurity place these infants at significant risk for bacterial invasion and systemic infection . Long-term sequelae include, but are not limited to, prolonged hospital stays and increased risk of neurodevelopmental problems . Nurses must understand the function of the immune system and the disadvantages unique to newborn infants to be able to provide education to others and appreciate the need for vigilant monitoring and protection of these infants.

PDA J Pharm Sci Technol, 2001 Nov-Dec, 55(6), 337 - 45
A method for the rapid detection of microbial contaminants in animal cell culture processes; Onadipe A et al.; The early detection of microbial contamination is an important issue in the production of biopharmaceuticals using animal cell culture systems . A new method, based on the ChemScan RDI analyser, was evaluated for the rapid detection and enumeration of low concentrations of microorganisms within a large population of animal cells and in cell culture media . The method was tested with suspension cell cultures and is applicable to adherent cell cultures . In both cases, the method uses an initial step to eliminate the animal cells followed by collection and fluorescent labelling of the viable microbial contaminants on a filter membrane . Total counts of the viable microorganisms were obtained after analysis of the membrane by the ChemScan RDI analyser . The results showed that the ChemScan RDI detected individual bacterial cells after filtration of the pre-treated animal cell culture . The detection limit of the ChemScan RDI was less than 10 bacteria/ml in cell culture containing 10(6) mammalian cells/ml and one bacterium in 500 ml of cell culture medium . A strong correlation between the standard plate count and the ChemScan RDI was observed, even at low bacterial concentrations . The total time for each analysis was less than two hours.

Mikrobiologiia, 2001 Sep-Oct, 70(5), 656 - 61
{Lysozyme-antilysozyme interactions in protozoa-bacteria communities (a model Tetrahymena-Escherichia community)}; Bukharin OV et al.; Lysozyme and antilysozyme activities present in a wide range of microorganisms determine the so-called lysozyme-antilysozyme system of hydrobionts, which greatly contribute to the formation of aquatic biocenoses . However, the mechanism of the functioning of this system in natural freshwater communities remains obscure . The experimental investigation of lysozyme-antilysozyme interactions in a model Tetrahymena--Escherichia community showed that the antilysozyme activity of Escherichia coli leads to incomplete phagocytosis, thus enhancing bacterial survival in a mixed culture with infusoria . The selection and reproduction of bacterial cells resistant to grazing by infusoria determine the character of host-parasite interactions and allow bacteria to survive . It was demonstrated that the antilysozyme activity of microorganisms, which is responsible for bacterial persistency in natural biocenoses, is involved in the maintenance of protozoa-bacteria communities in bodies of water.

Res Microbiol, 2001 Nov, 152(9), 767 - 70
Smart behavior of true slime mold in a labyrinth; Nakagaki T; Even for humans it is not easy to solve a maze . But the plasmodium of true slime mold, an amoeba-like unicellular organism, has shown an amazing ability to do so . This implies that an algorithm and a high computing capacity are included in the unicellular organism . In this report, we discuss information processing in the microorganism to focus on the issue as to whether the maze-solving behavior is akin to primitive intelligence.

Microbiol Immunol, 2001, 45(10), 721 - 7
Characterization of two genes encoding ferritin-like protein in Actinobacillus actinomycetemcomitans; Hirosue M et al.; Two genes encoding ferritin-like protein, designated afnA and afnB, were identified in the upstream region of actX on the Actinobacillus actinomycetemcomitans chromosomal DNA . The actX has been reported to be a regulatory gene homologous to the Escherichia coli fnr, which controls the growth and virulence of A . actinomycetemcomitans under anaerobic conditions . The afnB located 340 bp-upstream from the actX, and the afnA located just 15 bp-upstream from afnB . The afnA and afnB encoded 161 and 165 amino acid residues, respectively, which were similar to ferritin-like proteins of other microorganisms . Western immunoblotting using rabbit antiserum against E . coli ferritin showed these two proteins, which are reactive with the serum with 19-kDa molecular masses, are produced from A . actinomycetemcomitans . The N-terminal amino acid sequences of the two proteins were consequent with those deduced from afnA and afnB . Northern hybridization revealed that the afnA and afnB constituted a bicistronic operon and the accumulation of afnA and afnB mRNA was upregulated under aerobic conditions . These findings suggested that the operon was regulated by the presence of oxygen . The two ferritin-like proteins may have important roles in the adaptation of A . actinomycetemcomitans to oxidative environmental changes.

Appl Microbiol Biotechnol, 2001 Nov, 57(4), 474 - 81
Biotechnological development of effective phytases for mineral nutrition and environmental protection; Lei XG et al.; Phytases are hydrolytic enzymes that initiate the release of phosphate from phytate (myo-inositol hexakisphosphate), the major phosphorus (P) form in animal feeds of plant origin . These enzymes can be supplemented in diets for food animals to improve P nutrition and to reduce P pollution of animal excreta . This mini-review provides a synopsis of the concept of "ideal phytase" and the biotechnological approaches for developing such an enzyme . Examples of Escherichia coli AppA and Aspergillus fumigatus PhyA are presented to illustrate how new phytases are identified from microorganisms and developed by genetic engineering based on the gene sequences and protein structures of these enzymes . We also discuss the characteristics of different heterologous phytase expression systems, including those of plants, bacteria, fungi, and yeast.

Orv Hetil, 2001 Oct 28, 142(43), 2377 - 80
{Biological effects of microorganisms, their substances and immunosuppressive agents in mice}; Anderlik P; The author provide evidence from several aspects that the normal microbial flora has a permanent and life-long immune modulating role in conventional organisms and a stimulating effect both on specific and non-specific defence . However, in case of artificial interventions (stress, drugs) affecting the organism, existence of the normal flora may have an adverse effect (endotoxin effect, bacterial translocation) . The immunomodulants show a stimulating affect mainly in organisms with undeveloped immune system, and their effects are independent from the presence or absence of the microbial flora . With ageing, effect of immunomodulants can change and become indifferent or even suppressive . Dose-dependence of stimulating or suppressing effect of immunomodulants may be related to their non-immunological effects (endotoxin effect, bacterial translocation) . Finally, on the basis of the results, the authors consider the germfree mouse suitable for examining the effect of a given agent in the practice, on one hand, and for observing the host organism's reactions, free from the influence of the normal microbial flora, on the other . Along with the known physiological and pathological events, the results draw also attention to as distant fields as drug sensitivity, drug interactions influencing drug sensitivity . The author put emphasis on importance of germfree environment during immunosuppressive treatments in humans and when making special examinations under experimental conditions.

J Chemother, 2001 Oct, 13(5), 473 - 93
Preclinical and clinical pharmacology of biotherapeutic agents: Saccharomyces boulardii; Periti P et al.; Probiotic agents are living microorganisms that, upon ingestion, exert health benefits beyond inherent general nutrition . In this context, we must differentiate between biotherapeutics as approved drugs and dietary supplements and food products containing probiotic bacteria that are not considered drugs . At present the only biotherapeutic agent which is prescribable in some European countries, indicated to relieve specific diseases, is the yeast Saccharomyces boulardii . In this review we consider the various preclinical and clinical aspects of biotherapeutics as basic drugs and the biotherapeutic powers of their use in the treatment of some surgical enteropathies.

Int J Food Microbiol, 2001 Oct 22, 70(1-2), 53 - 61
Biological control of postharvest pear diseases using a bacterium, Pantoea agglomerans CPA-2; Nunes C et al.; Epiphytic microorganisms isolated from the fruits and leaf surfaces of apples and pears were screened for antagonistic activity against Penicillium expansum on pears . From 247 microorganisms tested for antagonistic properties against P . expansum, a bacterium strain identified as Pantoea agglomerans (CPA-2) was selected . This bacterium was very effective against Botrytis cinerea, P . expansum and Rhizopus stolonifer . Complete control at the three tested concentrations (2 x 10(7), 8 x 10(7) and 1 x 10(8) CFU ml(-1)) was obtained on wounded pears inoculated with 10(3), 10(4) and 10(5) conidia ml(-1) of P . expansum and R . stolonifer . At 8 x 10(7) CFU ml(-1), Pan . agglomerans reduced B . cinerea decay by more than 80% at the three concentrations of the pathogen . In over 3 years of experiments in semicommercial trials, Pan . agglomerans provided excellent control against B . cinerea and P . expansum under cold storage, either in air or in low oxygen atmospheres . Equal control was obtained with Pan . agglomerans at 8 x 10(7) CFU ml(-1), as with the fungicide imazalil at commercial doses, against both pathogens . Pan . agglomerans grew well inside wounds on pears at both room and cold temperatures and under modified atmospheres . In contrast, it grew poorly on the surface of intact fruit.

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