Environ Sci Technol, 2001 May 1, 35(9), 1785 - 90 Anaerobic degradation of methyl tert-butyl ether (MTBE) and tert-butyl alcohol (TBA); Finneran KT et al.; The potential for anaerobic degradation of methyl tert-butyl ether (MTBE) and tert-butyl alcohol (TBA) was investigated in laboratory incubations of sediments from a petroleum-contaminated aquifer and in aquatic sediments . The addition of humic substances (HS) stimulated the anaerobic degradation of MTBE in aquifer sediments in which Fe(III) was available as an electron acceptor . This is attributed to the fact that HS and other extracellular quinones can stimulate the activity of Fe(III)-reducing microorganisms by acting as an electron shuttle between Fe(III)-reducing microorganisms and insoluble Fe(III) oxides . MTBE was not degraded in aquifer sediments without Fe(III) and HS . {14C}-MTBE added to aquatic sediments adapted for anaerobic MTBE degradation was converted to 14CO2 in the presence or absence of HS or the HS analog, anthraquione-2,6-disulfonate . Unamended aquatic sediments produced 14CH4 as well as 14CO2 from {14C}-MTBE . The aquatic sediments also rapidly consumed TBA under anaerobic conditions and converted {14C}-TBA to 14CH4 and 14CO2 . An adaptation period of ca . 250-300 days was required prior to the most rapid anaerobic MTBE degradation in both sediment types, whereas TBA was metabolized in the aquatic sediments without a lag . These results demonstrate that, under the appropriate conditions, MTBE and TBA can be degraded in the absence of oxygen . This suggests that it may be possible to design strategies for the anaerobic remediation of MTBE in petroleum-contaminated subsurface environments. Am J Physiol Regul Integr Comp Physiol, 2001 Jun, 280(6), R1897 - 901 Influence of barometric pressure on interleukin-1 beta secretion; Becker WJ et al.; Monocytes and macrophages are activated by various environmental challenges, including microorganisms, radiation, and pollutants . These cells release cytokines, such as interleukin (IL)-1 beta, that mediate physiological adaptations to stress . This study sought to define further the role of IL-1 beta in general adaptation to environmental stress by testing the hypothesis that high altitude (20,000 ft, 6,096 m) would stimulate IL-1 beta secretion from isolated human blood mononuclear cells . Cells from six young men (aged 22--26 yr) were divided into separate cultures incubated in either standard ambient conditions or in one of three test conditions, hypobaric hypoxia (simulating 20,000 ft), hypobaric normoxia (20,000 ft, O(2) supplemented), and normobaric hypoxia (10% O(2)) . This design allowed differentiation between pressure-related vs . oxygen-related effects . Each subject made multiple blood donations in order that cells from all subjects were tested in all conditions . Contrary to the hypothesis, IL-1 beta secretion was not induced at simulated altitude in basal cell cultures . In lipopolysaccharide-stimulated cell cultures, exposure to altitude inhibited IL-1 beta secretion by approximately 40%, and the inhibition was due to the change in pressure (P = 0.039) rather than the change in oxygen . Secretion of other factors (IL-1 receptor antagonist and soluble IL-1 receptor type II) was not inhibited . Although these results are in opposition to the original hypothesis, they provide insight regarding adaptations necessary for hematopoiesis in response to high altitude and also provide a cellular rationale for the mountain sanatoriums of the 19th and early 20th centuries. Acc Chem Res, 2001 May, 34(5), 399 - 408 Enzymology, structure, and dynamics of acetohydroxy acid isomeroreductase; Dumas R et al.; Acetohydroxy acid isomeroreductase is a key enzyme involved in the biosynthetic pathway of the amino acids isoleucine, valine, and leucine . This enzyme is of great interest in agrochemical research because it is present only in plants and microorganisms, making it a potential target for specific herbicides and fungicides . Moreover, it catalyzes an unusual two-step reaction that is of great fundamental interest . With a view to characterizing both the mechanism of inhibition by potential herbicides and the complex reaction mechanism, various techniques of enzymology, molecular biology, mass spectrometry, X-ray crystallography, and theoretical simulation have been used . The results and conclusions of these studies are described briefly in this paper. Acc Chem Res, 2001 May, 34(5), 339 - 49 The central enzymes of the aspartate family of amino acid biosynthesis; Viola RE; The aspartate pathway is responsible for the biosynthesis of lysine, threonine, isoleucine, and methionine in most plants and microorganisms . The absence of this pathway in humans and animals makes the central enzymes potential targets for inhibition, with the aim of developing new herbicides and biocides, and also for enhancement, to improve the nutritional value of crops . Our current state of knowledge of these enzymes is reviewed, including recently determined structural information and newly constructed bifunctional fusion enzymes. Science, 2001 May 11, 292(5519), 1119 - 22 Factors that alter rumen microbial ecology; Russell JB et al.; Ruminant animals and ruminal microorganisms have a symbiotic relationship that facilitates fiber digestion, but domestic ruminants in developed countries are often fed an abundance of grain and little fiber . When ruminants are fed fiber-deficient rations, physiological mechanisms of homeostasis are disrupted, ruminal pH declines, microbial ecology is altered, and the animal becomes more susceptible to metabolic disorders and, in some cases, infectious disease . Some disorders can be counteracted by feed additives (for example, antibiotics and buffers), but these additives can alter the composition of the ruminal ecosystem even further. Environ Sci Technol, 2001 Mar 1, 35(5), 894 - 900 Metal speciation dynamics and bioavailability . 2 . Radial diffusion effects in the microorganism range; Pinheiro JP et al.; The free ion activity model for the biouptake of metals from complex media is limited to cases where mass transfer is not flux determining . This paper follows a previous paper (Van Leeuwen, H . P . Environ . Sci . Technol . 1999, 33, 3743) where speciation dynamics and bioavailability of metals are analyzed in terms of bioconversion kinetics and simultaneous metal transport in the medium coupled with dissociation kinetics . Such analysis shows under what conditions labile complex species contribute to the biouptake process or, equivalently, under what conditions the free ion activity model is not obeyed . The present work addresses the theoretical extension of the expressions for the metal flux in the medium by a radial diffusion term so that these are also applicable in the microorganism size range . The transition from macroscopic to microscopic surfaces affects not only the nature of the flux but also the extent of lability of complex species (Van Leeuwen, H . P.; Pinheiro, J . P . J . Electroanal . Chem . 1999, 471, 55), and this can have a dramatic influence on the rate of biouptake of metal ions . Labilities of metal complexes and the ensuing limiting metal fluxes are therefore systematically analyzed for various dimensions of the uptaking surface . Different conditions of bioaffinities and bioconversion capacities are considered, and a number of examples of metal complexes with specified kinetic features are discussed. Environ Toxicol Chem, 2001 Jan, 20(1), 84 - 98 Toxicity of pesticides to aquatic microorganisms: a review; DeLorenzo ME et al.; Microorganisms contribute significantly to primary production, nutrient cycling, and decomposition in estuarine eco-systems; therefore, detrimental effects of pesticides on microbial species may have subsequent impacts on higher trophic levels . Pesticides may affect estuarine microorganisms via spills, runoff, and drift . Both the structure and the function of microbial communities may be impaired by pesticide toxicity . Pesticides may also be metabolized or bioaccumulated by microorganisms . Mechanisms of toxicity vary, depending on the type of pesticide and the microbial species exposed . Herbicides are generally most toxic to phototrophic microorganisms, exhibiting toxicity by disrupting photosynthesis . Atrazine is the most widely used and most extensively studied herbicide . Toxic effects of organophosphate and organochlorine insecticides on microbial species have also been demonstrated, although their mechanisms of toxicity in such nontarget species remain unclear . There is a great deal of variability in the toxicity of even a single pesticide among microbial species . When attempting to predict the toxicity of pesticides in estuarine ecosystems, effects of pesticide mixtures and interactions with nutrients should be considered . The toxicity of pesticides to aquatic microorganisms, especially bacteria and protozoa, is an area of research requiring further study. Ann Rheum Dis, 2001 Jun, 60(6), 632 - 4 Intra-articular co-infection by Borrelia burgdorferi and Chlamydia trachomatis; Putschky N et al.; OBJECTIVE: Chlamydia trachomatis and Borrelia burgdorferi infections are frequently the cause of unexplained oligoarthritis, as shown by identification of bacteria specific DNA in joint material from patients with reactive arthritis, Lyme arthritis, and undifferentiated oligoarthritis . The aim of this study was to determine whether the two organisms occur simultaneously in joint material from patients with arthritis . METHODS: Seventy six patients with unexplained arthritis were prospectively studied . Synovial fluid was obtained from all patients and examined for DNA from C trachomatis and B burgdorferi using specific polymerase chain reaction (PCR) protocols . Data concerning prior genitourinary infection or a history of tick bite were recorded and serum antibodies to C trachomatis and B burgdorferi were determined . RESULTS: Six patients (8%) had DNA from both C trachomatis and B burgdorferi in the same synovial fluid specimen (mean leucocyte count 11.925/mm(3), 65% granulocytes) . These patients (four men, two women; mean age 33.7 years) all had oligoarthritis of the knee, ankle, or both (mean disease duration 11.3 months) . From the history and serological examination, four patients had some evidence of actual or previous infection with one or other of the bacteria, while the other two patients had a positive serological test for Chlamydia only . CONCLUSIONS: DNA from two different microorganisms which are known to be triggering agents for arthritis may be present simultaneously in joint material from patients with unexplained oligoarthritis . This finding raises the question as to whether, in such cases, one or both bacteria contribute to the pathogenesis of the disease or whether they are only innocent bystanders. Przegl Epidemiol, 2000, 54(3-4), 367 - 74 {Exposure to toxocariasis and tularemia among workers at Białowieza National Park}; Lapinski TW et al.; Everyday contact of the workers of Bialowieza sanctuary with animals may create a risk of infection by microorganisms attacking animals . The reported study was undertaken for carrying out an epidemiological analysis of tularemia and toxocarosis in this group of workers . Tularaemia infection was not found in then . In 12% of these workers antibodies to Toxocara canis were found, and fire workers had articular borreliosis . Antibodies to taxocara were disclosed in 10% of the Bialowieza population not employed in the sanctuary and 30% of the population of the village Kruklanka in the Pisz Forests . The workers of the Bialowieza Sanctuary are not exposed to Toxocara canis infection more than the remaining population of Bialowieza and the population in the Kruklanka village in a region far from Bialowieza. Environ Sci Technol, 2001 Feb 15, 35(4), 682 - 9 Signature metabolites attesting to the in situ attenuation of alkylbenzenes in anaerobic environments; Elshahed MS et al.; Accurate assessment of the fate of hydrocarbons spilt in aquifers is essential for gauging associated health and ecological risks . Regulatory pressure to actively remediate such contaminated ecosystems can be substantially diminished if solid evidence for in situ microbial destruction of pollutants is obtained . In laboratory incubations, sediment-associated microorganisms from a gas condensate-contaminated aquifer anaerobically biodegraded toluene, ethylbenzene, xylene, and toluic acid isomers with stoichiometric amounts of sulfate consumed or methane produced . The activation of the alkylated aromatic contaminants involved conversion to their corresponding benzylsuccinic acid derivatives, a reaction known to occur for toluene and m-xylene decay, but one previously unrecognized for ethylbenzene, o- and p-xylene, and m-toluate metabolism . Benzylsuccinates were further biodegraded to toluates, phthalates, and benzoate . In laboratory incubations, these metabolites were transiently produced . Several of the metabolites were also detected in groundwater samples from an aquifer where alkylbenzene concentrations decreased over time, suggesting that anaerobic microbial metabolism of these contaminants also occurs in situ . Our studies confirm the utility of the aforementioned compounds as signature metabolites attesting to the natural attenuation of aromatic hydrocarbons in anaerobic environments. Environ Sci Technol, 2001 Feb 15, 35(4), 658 - 62 Widespread potential for microbial MTBE degradation in surface-water sediments; Bradley PM et al.; Microorganisms indigenous to stream and lake bed sediments, collected from 11 sites throughout the United States, demonstrated significant mineralization of the fuel oxygenate, methyl-tert-butyl ether (MTBE) . Mineralization of {U-14C}MTBE to 14CO2 ranged from 15 to 66% over 50 days and did not differ significantly between sediments collected from MTBE contaminated sites and from sites with no history of MTBE exposure . This result suggests that even the microbial communities indigenous to newly contaminated surface water systems will exhibit some innate ability to attenuate MTBE under aerobic conditions . The magnitude of MTBE mineralization was related to the sediment grain size distribution . A pronounced, inverse correlation (p < 0.001; r2 = 0.73) was observed between the final recovery of 14CO2 and the percentage content of silt and clay sized grains (grain diameter < 0.125 mm) . The results of this study indicate that the microorganisms that inhabit the bed sediments of streams and lakes can degrade MTBE efficiently and that this capability is widespread in the environment . Thus aerobic bed sediment microbial processes may provide a significant environmental sink for MTBE in surface water systems throughout the United States and may contribute to the reported transience of MTBE in some surface waters. Infect Immun, 2001 Jun, 69(6), 3562 - 8 Mycobacterium bovis BCG recA deletion mutant shows increased susceptibility to DNA-damaging agents but wild-type survival in a mouse infection model; Sander P et al.; Pathogenic microorganisms possess antioxidant defense mechanisms for protection from reactive oxygen metabolites which are generated during the respiratory burst of phagocytic cells . These defense mechanisms include enzymes such as catalase, which detoxifies reactive oxygen species, and DNA repair systems, which repair damage resulting from oxidative stress . To (i) determine the relative importance of the DNA repair system when oxidative stress is encountered by the Mycobacterium tuberculosis complex during infection of the host and to (ii) provide improved mycobacterial hosts as live carriers to express foreign antigens, the recA locus was inactivated by allelic exchange in Mycobacterium bovis BCG . The recA mutants are sensitive to DNA-damaging agents and show increased susceptibility to metronidazole, the first lead compound active against the dormant M . tuberculosis complex . Surprisingly, the recA genotype does not affect the in vitro dormancy response, nor does the defect in the DNA repair system lead to attenuation as determined in a mouse infection model . The recA mutants will be a valuable tool for further development of BCG as an antigen delivery system to express foreign antigens and as a source of a genetically stable vaccine against tuberculosis. Environ Sci Technol, 2001 Jan 15, 35(2), 354 - 61 Kinetics of mass transfer-limited bacterial growth on solid PAHs; Wick LY et al.; The importance of mass transfer relative to the intrinsic microbial activity was examined in a laboratory system using Mycobacterium sp . LB501T and poorly soluble anthracene as sole carbon source . M . sp . LB501T was grown on various amounts of solid anthracene in batch cultures, and microbial biomass formation was compared to independently determined dissolution fluxes . Provision of only a few anthracene crystals (< or = 2 g L(-1)) resulted in pseudolinear growth due to low dissolution fluxes, whereas exponential growth was only obtained when high amounts of solid anthracene (30 g L(-1)) were provided . The influence of substrate bioavailability on microbial growth was predicted successfully by a dynamic, flux-based approach (Best-Equation), which combines substrate dissolution from crystals into solution, substrate uptake by microorganisms from solution, and concurrent biomass formation. Biomed Sci Instrum, 2001, 37, 191 - 6 Real time biodetection of individual pathogenic microorganisms in food and water; Johnson PE et al.; The primary objective of this research is to examine the feasibility of using an innovative technique based on laser-induced fluorescence coupled with flow cytometry to detect pathogenic microorganisms in food or water in real time . Our initial application is the rapid detection of E . coli O157:H7 in ground beef . The research performed demonstrated conclusively that this approach is feasible, and that the technique has key advantages over current alternatives including: it is (1) able to totally examine a large volume of food or water in real time, (2) capable of detecting single microorganisms (alternative techniques require in excess of 10(4) microorganisms), (3) intrinsically automatic, and (4) sensitive only to the selected bacteria . We have demonstrated the feasibility of detecting individual E . coli bacteria with a breadboard system . The performance of this system allows for rapid detection of individual specific pathogenic microorganisms . Two of the most significant commercial applications of this technique are the detection of infectious microorganisms in contaminated food and water . Food-borne microbial pathogens account for approximately 7 million illnesses and 9,000 deaths in the U.S . annually, with an estimated economic loss of at least $6 billion {1} . In addition, this method has the potential for a broad range of other commercial applications, including the detection of small numbers of molecules, such as the ultrasensitive detection of explosives and groundwater contaminants. Environ Technol, 2001 Mar, 22(3), 339 - 44 Quantitative bioregeneration of granular activated carbon loaded with phenol and 2,4-dichlorophenol; Vinitnantharat S et al.; Lignite based granular activated carbon of 20x30 mesh size was used to investigate the extent of bioregeneration of phenol and 2,4-DCP in a batch system . The adsorption isotherm fits very well with the Freundlich isotherm and it is apparent that 2,4-DCP is more adsorbable than phenol . The degree of reversible adsorption for phenol and 2,4-DCP were 32.9 and 10.6 %, respectively . The low percentages of their reversibility meant that the adsorption phenomena of both phenol and 2,4-DCP were not fully physicosorption . The microorganisms can regenerate 31.4% (21.2 mg g(-1)) of GAC loaded with phenol and 14.3% (24.8 mg g(-1)) of GAC loaded with 2,4-DCP over a period of 7-10 days . The bioregeneration of phenols closely followed the first order kinetics with the rate constant of 0.046 day(-1) at initial phenol concentration of 100 mg l(-1) and biomass concentration of 20 mg l(-1) MLVSS . The rate constant was 0.021 day(-1) at initial 2,4-DCP concentration of 200 mg l(-1) and 200 mg l(-1) MLVSS . In addition, the increasing in initial concentration of biomass in the solution shortened the time required to reach the asymptotic limit on the bioregeneration but rendered little impact on the bioregeneration percentage. Environ Toxicol Chem, 2001 Apr, 20(4), 721 - 6 Kinetics of polychlorinated biphenyl dechlorination and growth of dechlorinating microorganisms; Rhee GY et al.; The present study has investigated a correlation between the kinetics of polychlorinated biphenyl (PCB) dechlorination and the growth of dechlorinating microbial populations . Microorganisms were eluted from Aroclor 1248-contaminated St . Lawrence River (NY, USA) sediments and inoculated into clean sediments spiked with Aroclor 1248 at 10 concentrations ranging from 0 to 3.12 micromol/g sediment (0-900 ppm) . The time course of PCB dechlorination and population growth were concurrently determined by congener-specific analysis and the most probable number technique, respectively . The specific growth rate was a saturation function of PCB concentrations above the threshold concentration (0.14 micromol/g sediment, or 40 ppm), below which no dechlorination or growth of dechlorinations were observed . The maximum growth rate was 0.20/d with a half-saturation constant of 1.23 micromol/g sediment . The yield of dechlorinating microorganisms showed a peak at 0.70 micromol/g sediment (200 ppm), with a value of 10.3 x 10(12) cells/mol Cl removed, and decreased below and above this concentration . The dechlorination rate (micromol Cl removed/g sediment/d) was a linear function of Aroclor concentration . Both the log of this rate and the maximum level of dechlorination were significantly correlated with growth rate . The biomass-normalized dechlorination rate (micromol Cl removed/g sediment/cell/d) was first order because of the exponential manner of the population growth . The first-order rate constant was a saturation function of Aroclor concentrations, with a maximum of 0.24/d (a half-life of 2.9 d) and a half-saturation constant of 1.18 micromol/g sediment, which are similar to the constants for growth . These results indicate that the dechlorination rate is tightly linked to the population growth of dechlorinating microorganisms. J Clin Invest, 1974 Feb, 53(2), 343 - 53 Serum heat-labile opsonins in systemic lupus erythematosus; Jasin HE et al.; To study possible mechanisms responsible for the increased susceptibility to infection of patients with active systemic lupus erythematosus (SLE), a study of the serum heat-labile opsonic capacity (HLOC) in such patients was undertaken . With leukocytes from normal donors, the sera of 12 of 30 patients with active SLE demonstrated decreased HLOC for E . coli 075 . The phagocytic activity was partially restored by normal serum, suggesting that decreased HLOC was responsible for the defective phagocytosis . While 8 of 10 patients with active SLE and concomitant infections showed deficient opsonic capacity to E . coli 075, only 4 of 20 such patients without infections showed the defect (P = 0.01) . None of 12 patients with inactive disease had deficient opsonic capacity . Similar results were obtained with S . aureus 502A as the test bacterium . In the patients surviving infection, recovery of normal serum opsonic capacity was rapid and usually coincided with an increase of serum complement to normal levels . In three patients with active SLE and infection, the causative microorganisms were isolated and opsonic capacity for these organisms tested with the individual patients' sera . In each case, sera obtained at the onset of the infectious episode had low opsonic capacity when compared with normal sera . Serum C3 proactivator levels were low in 9 of 11 sera with deficient opsonic capacity . However, similar low values were found in other SLE sera with normal HLOC, suggesting that other factors of the opsonic system were also depleted . Addition of the classical complement components C1, C4, C2, C3, and C5 to sera with deficient HLOC failed to restore activity . Addition of pure C3 proactivator also failed to restore activity . However, addition of C3 proactivator together with 50 degrees C-heated normal serum restored activity, indicating that factors active at the early steps of opsonic activation via the alternative pathway of complement were necessary to restore opsonic activity . These findings indicate that in active SLE, a decrease of components of the alternate pathway of complement activation results in an acquired defect of serum HLOC and perhaps other related complement-mediated functions . This defect may be an important factor in the increased susceptibility to infections of patients with active systemic lupus erythematosus. J Allergy Clin Immunol, 2001 May, 107(5), 856 - 62 Group 13 grass allergens: structural variability between different grass species and analysis of proteolytic stability; Petersen A et al.; BACKGROUND: Determination of the allergen composition of an extract is essential for the improvement of hyposensitization therapy . Surprisingly, although grass pollen extracts have been studied intensively for 20 years, a further major allergen, Phl p 13, was detected recently in timothy grass pollen . OBJECTIVES: We sought to determine the occurrence and importance of group 13 allergens in various grass species and to investigate their proteolytic stability . METHODS: The group 13 allergens were determined by means of 2-dimensional PAGE blotting with patient sera and group 13-specific mAbs . The allergens were isolated chromatographically from several pollen extracts and analyzed by means of microsequencing . Cross-reactivity among various grass species was studied by using Western blots and immunoblot inhibition tests . The stability of the allergens was tested under defined extraction conditions . RESULTS: Group 13 allergens are detectable in all common grasses and show IgE cross-reactivity among them . The allergenic components were identified in the neutral pH range with molecular masses of 50 to 60 kd, and in the case of Phl p 13, maximal binding of the isoforms was observed at 55 kd and at an isoelectric point of 6 to 7.5 . Protein sequencing clearly confirms structural identities between different grass species, although individual variations are found . If low-molecular-mass components were depleted by means of gel filtration, a rapid degradation of group 13 allergens was observed . This is in contrast to other pollen allergens described thus far . CONCLUSION: Group 13 allergens are widespread and are major allergens in the grasses . Predicted from their primary structures, these allergens are polygalacturonases . This class of enzymes is already known from microorganisms, and these enzymes are recognized as potential inducers of asthma . Our studies indicate that the group 13 allergens show a considerable microheterogeneity and degradation, especially after depletion of low-molecular-mass components . One has to be aware of this pivotal fact when soluble grass pollen extracts are prepared for diagnostics and hyposensitization therapy. Dig Dis Sci, 2001 May, 46(5), 1127 - 32 Acute pancreatitis and bacterial translocation; Cicalese L et al.; Infectious complications are the most frequent and severe complications of acute narcotizing pancreatitis (AP) with a mortality rate up to 80% . Although experimental and clinical studies suggest that the microbiologic source of pancreatic infection could be enteric, information in this regard is scant . This study evaluated bacterial translocation (BT) using mild and severe models of AP . Mild AP was induced by 6-hr continuous intravenous infusion of cerulein, while severe AP was induced by additional infusion of glycodeoxycholic acid into the biliopancreatic duct . BT was evaluated with organ cultures performed when animals were killed (24 hr) . To confirm the gastrointestinal origin of the translocating microorganisms, fluorescent microspheres were also given to the animals in drinking water 24 hr before induction of AP . At the time of death beads were counted with a (fluorescence-activated cell sorter) (FACS) in peritoneal lavages and with fluorescent microscopy in frozen sections of the pancreata . Morphology of the distal small bowel showed significant changes in the animals with AP compared to controls, such as reduction of villus high and altered microvasculature . Mild AP induced BT to the pancreas in 100% of the animals, compared to pancreata from control groups . Severe AP induced increased BT to the pancreas . BT to liver and spleen was also significantly increased with AP . The presence of fluorescent microspheres confirmed their enteric derivation . This study provides evidence for the enteric origin of microorganisms responsible for pancreatic infectious complications during AP . The evidence of BT after laparotomy suggests an increased risk of infections with the association of these conditions . This could provide an explanation for the high mortality associated with laparotomy in course of AP. Appl Microbiol Biotechnol, 2001 Apr, 55(3), 290 - 5 Production of 3-nitrocatechol by oxygenase-containing bacteria: optimization of the nitrobenzene biotransformation by Nocardia S3; Kieboom J et al.; Twenty-one microorganisms were screened for their ability to convert nitroaromatics into 3-nitrocatechol as a result of the action of an oxygenase . Cultures containing toluene dioxygenases and phenol monooxygenases accumulated 3-nitrocatechol during incubation with nitrobenzene and nitrophenol, respectively . Nocardia S3 was selected and studied in more detail . Toluene-pregrown cultures were able to degrade nitrobenzene with a concomitant formation of 3-nitrocatechol . The rates of nitrobenzene utilization decreased throughout the biotransformation period and finally the accumulation ceased . The gradual deterioration of the biotransformation rates was not a consequence of depletion of the NADH pool, but was due to the accumulation of 3-nitrocatechol . The inhibition of nitrobenzene biotransformation by 3-nitrocatechol greatly impacts 3-nitrocatechol production processes. Ground Water, 2001 May-Jun, 39(3), 348 - 55 In situ determination of subsurface microbial enzyme kinetics; Istok JD et al.; The single-well, push-pull test has been used in previous field studies to measure in situ zero- and first-order rates for aerobic and anaerobic microbial respiration in the saturated zone . In this paper we demonstrate that the test can also be used to obtain more generalized descriptions of the kinetics of microbially mediated enzymatic reactions . Laboratory and field tests were performed with the model enzyme substrate p-nitrophenyl-beta-D-glucopyranoside (PNG) . During a push-pull test, injected PNG is hydrolyzed in situ to p-nitrophenol (PNP); the rate of PNP production is taken as a measure of the beta-glucosidase activity expressed by indigenous microorganisms . Laboratory tests were performed in physical aquifer models packed with natural aquifer sediment; field tests were performed in a shallow unconfined alluvial aquifer at a petroleum contaminated site . The laboratory and field tests demonstrate that it is possible to compute the in situ rate of PNP production as a function of PNG concentration using only data from a single push-pull test . These data can then be used to estimate the Michaelis-Menton kinetic parameters Vmax and Km for the hydrolysis reaction . This approach potentially extends the range of applicability of the push-pull test approach for use in determining kinetic parameters for a wide range of microbial processes in situ . These could include the broad class of substituted nitrophenyl substrates used to assay other enzyme systems, as well as microbially mediated redox reactions that occur during contaminant transformations. Clin Infect Dis, 2001 Jun 1, 32(11), 1589 - 601 Epub 2001 Apr 30. Infectious disease pathology; Procop GW et al.; The anatomic pathologist performs an important role in the diagnosis or exclusion of infectious diseases . The morphologic interpretation of biopsies and cytologic preparations allows for the definitive establishment or exclusion of a wide variety of diseases . Once the pathologist has determined that a disease is likely to be due to an infection and has characterized the inflammatory response, associated microorganisms or viral-associated cytopathic effects should be recorded . Although some microorganisms or their cytopathic effects may be clearly visible on routine hematoxylin and eosin-stained sections, additional histochemical stains are often needed for their complete characterization . Highly specific molecular techniques, such as immunohistochemistry, in situ hybridization, and nucleic acid amplification, may be needed in certain instances to establish the diagnosis of infection . Through appropriate morphologic diagnoses and interlaboratory communication and collaboration, the anatomic pathologist contributes greatly to the diagnosis and treatment of infectious diseases. Mikrobiologiia, 2001 Jan-Feb, 70(1), 106 - 10 {Effect of certain environmental factors on Bradyrhizobium japonicum}; Kurdish IK et al.; Dependence of motility and chemotaxis was studied in two strains of Bradyrhizobium japonicum upon several environmental factors . In both strains, chemotaxis was found to increase with an increasing concentration of the attractant (glucose) to 5.6 x 10(-2) M . Both motility and chemotaxis reached their maximum in the two- to three-day cultures at neutral pH . The maximum motility of these bacteria occurred at 40 degrees C . The maximum values of chemotaxis in these microorganisms were, however, observed at 20-25 degrees C . Chemotaxis in acidic or alkaline media and at low temperatures was found to be markedly weaker . Nonoptimal values of these parameters in soil may be a limiting factor for the interaction of the given bacteria with soybean roots. Am J Clin Pathol, 2000 Dec, 114(6), 880 - 9 Quantitative analysis of valvular lesions during Bartonella endocarditis; Lepidi H et al.; Cardiac valve pathology was evaluated in 15 patients with confirmed diagnosis of Bartonella endocarditis . Ten were infected by Bartonella quintana and 5 by Bartonella henselae . Histologic features of these cases, including fibrosis, calcification, vegetation, pattern of inflammation, and vascularization, were compared with those of valves from 25 cases of non-Bartonella endocarditis as controls using a computerized quantitative image analysis . Pathologic and immunohistologic testing for localization of Bartonella species in resected valves included Warthin-Starry stain and polyclonal antibody-based immunodetection . Compared with other cases of infective endocarditis, cases of Bartonella endocarditis are more fibrotic and calcified, less vascularized, with less extensive vegetation and chronic inflammation . These pathologic changes are suggestive of a prolonged infection . Warthin-Starry stain and immunohistologic testing demonstrated the presence of the organism, respectively, in 11 and 10 of the 13 tested valves . Results of both staining methods showed microorganisms in extracellular locations and in regions unaccompanied by inflammation . Pathology and immunohistology may contribute to the etiologic diagnosis of Bartonella endocarditis when serology and molecular techniques are not available. Vestn Ross Akad Med Nauk, 2001, (3), 18 - 21 {Cold shock in bacteria}; Basnak'ian IA; Various species and genera of bacterial show the same responses to low temperature . Cold induces cold shock genes, by causing significant changes in the regulation of protein synthesis . The synthesis of major proteins in the microbial cell is suppressed . However, there is synthesis of a great deal of new proteins, the so-called cold shock ones . The chief protein in this family is E . coli CspA that activates the translation of other cold shock genes and negatively regulates the expression of its own gene . E . coli CspA homologies were identified in many bacteria . They can be also identified in other microorganisms, including the pathogens of infectious diseases . This can be attributable to the presence of common antigens in different bacteria . The data presented should be borne in mind in identifying bacteria and in designing immunodiagnostic agents if there is a culture cooling stage in their technology. Ann Clin Lab Sci, 2001 Apr, 31(2), 157 - 61 Cytomegalovirus seropositivity and serum total cholesterol levels in young patients; Froberg MK et al.; Atherosclerosis is a chronic inflammatory disease of arteries, associated with multiple genetic and environmental factors, including hypertension, diabetes mellitus, cigarette smoking, modified and elevated LDL cholesterol, elevated plasma homocysteine, and infectious microorganisms such as Chlamydia pneumoniae and cytomegalovirus (CMV) . CMV has been implicated in atherogenesis by epidemiological studies, animal research, and molecular analyses that have demonstrated CMV nucleic acids within human atherosclerotic lesions . Studies have suggested that CMV infection may alter lipid metabolism and lead to accumulation of cholesterol within atheromatous plaques . Few studies have examined the relationship between CMV infection and serum cholesterol levels in younger individuals when much of atherogenesis occurs . To test if CMV-seropositivity is associated with high levels of serum total cholesterol in relatively young patients, CMV IgG levels and total cholesterol concentrations were analyzed in serums from 172 patients, age < 50 yr . Based on univariate analysis of variance, serum total cholesterol was significantly correlated to age and to CMV-seropositivity when gender was a cofactor, but not to gender or CMV-seropositivity alone . In 39 CMV-seropositive women, serum total cholesterol concentration averaged 218 +/- 50 mg/dL (mean +/- SD), which was significantly higher than in 53 CMV-seronegative women (194 +/- 39 mg/dL, p < 0.02) . No significant difference was observed between the serum total cholesterol concentrations in 26 CMV-seropositive men and 51 CMV-seronegative men (198 +/- 42 mg/dL versus 212 +/- 48 mg/dl, respectively) . Thus, this study provides evidence that CMV-seropositivity is associated with higher serum total cholesterol levels in female patients under 50 yr of age, but not in male patients of comparable age. Environ Toxicol Chem, 2001 May, 20(5), 1133 - 40 The relation between extrapolated risk, expressed as potentially affected fraction, and community effects, expressed as pollution-induced community tolerance; Van Beelen P et al.; The results of toxicity tests can be used to calculate the potentially affected fraction (PAF) of species in an ecosystem at a given pollutant concentration using statistical extrapolation methods . The PAF curve indicates the fraction of species from the original community that may become inhibited at each elevated pollutant concentration and is a measure of the ecotoxicological risk . Pollution-induced community tolerance (PICT) is a true community response that is measured under controlled conditions in the laboratory, using organisms from contaminated field sites . Microorganisms from experimental field plots with added Zn were exposed to various concentrations of Zn in the laboratory and the mineralization of 14C acetate was monitored . Microorganisms from plots with Zn concentrations above 124 mg/kg showed a significant increase in the effect concentration 10% (EC10) and, therefore, had a significant PICT . The pore-water concentrations of Zn in these field soils were in the same magnitude as the EC10 of the microorganisms from these soils . The PAF curve was calculated from previously reported toxicity tests with five different microbial species using the average and the standard deviation of the logarithmically transformed EC10 values . The average sensitivity of this PAF curve was similar to the EC50 of the acetate mineralization curve from the field plot without added Zn2+, but the PAF curve was less steep . Our experiments indicated that 27 to 84% of the original microbial species were inhibited at Zn concentrations from 334 to 1,858 mg/kg soil, respectively . Our results suggest that the PICT method can now also be used to quantify the fraction of the original species composition that is inhibited at a specific pollutant concentration. J Biol Chem, 2001 Jul 13, 276(28), 25894 - 902 Epub 2001 May 03. Stoichiometry of complexes between mannose-binding protein and its associated serine proteases . Defining functional units for complement activation; Chen CB et al.; Serum mannose-binding protein (MBP) initiates the lectin branch of the complement cascade by binding to sugars on the surfaces of microorganisms and activating two MBP-associated serine proteases (MASP-1 and MASP-2) . Rat serum MBP consists of oligomers containing up to four copies of a subunit that is composed of three identical polypeptide chains . Biophysical analysis of intact and truncated MASPs indicates that each MASP is a homodimer that is stabilized through interactions involving an N-terminal CUB domain . The binding sites for MBP are formed from the three N-terminal MASP domains, in which two CUB modules interact with MBP . Each MASP dimer contains binding sites for two MBP subunits . Both sites must be occupied by subunits from a single MBP oligomer to form a stable complex . Thus, the smallest functional unit for complement activation consists of MBP dimers bound to MASP-1 or MASP-2 homodimers . Trimers and tetramers of MBP form complexes containing up to two MASPs . The results reveal how MASP-1 and MASP-2 can function independently to activate the complement cascade. Diagn Microbiol Infect Dis, 2001 Mar, 39(3), 161 - 4 Trichosporon species infection in bone marrow transplanted patients; Moretti-Branchini ML et al.; Trichosporon species are emerging as opportunistic agents that cause systemic diseases in immunocompromised patients . Patients undergoing bone marrow transplant are submitted to intense and prolonged periods of neutropenia and consequently to several risk factors to fungal infections as the use of broad spectrum antibiotics and invasive devices . Two cases of fungal infections caused by Trichosporon asahii var . asahii and T . inkin in patients with bone marrow transplant are described T . asahii var . asahii was responsible for fungemia and the identification of this microorganism was later performed . T . inkin caused vascular accesses infection and was recovered from an implanted Hickman-Broviac catheter . Both patients were under oral fluconazole prophylaxis . The patient with systemic infection died despite the therapy with amphotericin B and the patient with catheter-related infection recovered from the fungal infection after catheter removal . Difficulties in the identification of this microorganism lead to delays in treatment and post-mortem diagnosis. Trends Microbiol, 2001 May, 9(5), 228 - 37 Maturation of the {NiFe} hydrogenases; Casalot L et al.; The high degree of similarity that exists between all the {NiFe} hydrogenase operons and the near universality of hydrogen metabolism among microorganisms suggest that the microbial ability to metabolize hydrogen is of great importance and ancient origin . The large number of genes present in these operons, which are mostly involved in the maturation of the structural subunit, is indicative of the complexity of the hydrogenase molecular structure . Two main groups of maturation genes can be differentiated based on the resulting phenotypes when mutated: the 'cis-genes', encoding narrow specificity proteins, are mainly located on the same transcription unit as the structural genes, and the 'trans-genes', encoding broad specificity proteins, are located on a different operon . The maturation of the large subunit starts with the formation of a complex with the chaperone HypC, which remains bound to the amino terminus throughout processing . The ligands CN and CO, which are derived from carbamoylphosphate, are then inserted via HypF and probably other accessory proteins . HypB is responsible for nickel atom delivery in a GTP-hydrolysis-dependent reaction . The last identified step in the large subunit maturation process is proteolytic cleavage at the carboxyl terminus . The possible roles of the other maturation proteins are also discussed. Med Clin (Barc), 2001 Feb 24, 116(7), 256 - 8 {Reduction of hematogenous infection in HIV positive patients following highly active antiretroviral therapy}; Luisa Pedro-Botet M et al.; BACKGROUND: Bloodstream infections have probably decreased in HIV patients since highly active antiretroviral therapy (HAART) usage . PATIENTS AND METHOD: Hematogenous infections, including mycobacteria and fungi in HIV patients, stratified into two periods (I: 1995-1996 and II: 1997-1998) were included to evaluate changes attributable to HAART . RESULTS: 226 episodes were attended (incidence: 38.8 in period I and 15.3 in period II; p < 0.01) . A significant decrease was observed in immunosuppression related microorganisms . HAART usage increased from 6.4% in period I to 31.2% in period II . Immunological parameters of HIV patients significantly improved in period II . CONCLUSIONS: Hematogenous infections have significantly decreased in HIV patients since the use of HAART. Gastroenterol Hepatol, 2001 Apr, 24(4), 186 - 90 {Infectious complications associated with liver transplantation: analysis of 104 patients}; Cubiella J et al.; OBJECTIVE: Infectious complications are the main cause of morbidity and mortality during the first year after liver transplantation . The aim of the present study was to determine the incidence, microorganisms and factors associated with the development of infectious complications . PATIENTS AND METHOD: Retrospective analysis of infectious episodes during the first year after transplantation in 104 patients undergoing transplantation between April 1995 and December 1996 . The various clinical variables related to the pre-transplant disease, the surgical intervention and post-transplant evolution were evaluated with the aim of identifying predictive factors for the development of bacterial infectious complications . RESULTS: During the first year, 51 patients (49%) presented 111 infectious episodes . The most frequent infections were bacterial (66%); 21% were cytomegalovirus infections and 22% were fungal . The incidence of bacterial infections was highest during the first month (80% of all infectious episodes in this period) . Two variables were independently associated with the development of bacterial infections in the first month following transplantation: prolonged ischemia of the graft (p = 0.002) and length of stay in the intensive care unit (p = 0.002) . Infectious complications caused 8 of the 11 deaths that occurred during the 1-year follow-up . Mortality associated with invasive fungal infections was 100% . CONCLUSIONS: Although the overall incidence of infections and associated mortality has decreased, it remains the main cause of mortality and morbidity in the first year after transplantation. Physiol Biochem Zool, 2001 May-Jun, 74(3), 398 - 403 Effect of age on seed digestion in parrots (Amazona aestiva); Vendramin-Gallo M et al.; The objective of this study was to compare the capacity of adult (more than 3 yr old) and young (less than 1 yr old) true parrots to digest seeds that are normally included in their diet in captivity, particularly soybean, sunflower, and corn . All the seeds were offered for 5 d with an interval of 15 d between different diets . The seeds of soybean and corn were boiled for 15 min and soaked in water at ambient temperature for 12 h before being fed to the birds . There were no differences in the digestibilities of crude protein and fats (ether extract) among animals, but the digestibilities of dry matter and crude fiber by the adult animals were higher than those of the young ones . The digestibility of carbohydrate (nitrogen-free extract) by adult birds was higher only for sunflower seeds . It is concluded that the capacity of parrots to digest fiber may change according to the age of the animal . Since the digestion of fiber depends on the action of microorganisms, these results suggest that the colonization of the gastrointestinal tract is delayed or very slow in young parrots. Appl Microbiol Biotechnol, 2001 Mar, 55(2), 219 - 25 Studies on the possibility of histidine biosynthesis from histodinol, imidazolepyruvic acid, imidazoleacetica acid, and imidazolelactic acid by mixed ruminal bacteria, protozoa, and their mixture in vitro; Wadud S et al.; The possibility of histidine (His) synthesis using a main biosynthetic pathway involving histidinol (HDL) and also the recycling capability of imidazolic compounds such as imidazolepyruvic acid (ImPA), imidazoleacetic acid (ImAA), and imidazolelactic acid (ImLA) to produce His were investigated using mixed ruminal bacteria (B), protozoa (P), and a mixture of both (BP) in an in vitro system . Rumen microorganisms were anaerobically incubated at 39 degrees C for 18 h with or without each substrate (2 mM) mentioned . His and other related compounds produced in both the supernatants and hydrolyzates of the incubation were analyzed by high-performance liquid chromatography . B, P, and BP suspensions failed to show His synthesizing ability when incubated with HDL . His was synthesized from ImPA by B, P, and BP . Expressed in units "per gram of microbial nitrogen (MN)", ImPA disappearance was greatest in B (72.7 micromol/g MN per hour), followed by BP (33.13 micromol/g MN per hour) and then P (18.6 micromol/g MN per hour) for the 18-h incubation period . The production of His from ImPA in B (240.0, 275.9, and 261.2 micromol/g MN in 6, 12, and 18 h incubation, respectively) was about 3.5 times higher than that in P (67.3, 83.8, and 72.7 micromol/g MN in 6, 12, and 18 h incubation, respectively) . Other metabolites produced from ImPA were ImLA, ImAA, histamine (HTM), and urocanic acid (URA), found in all microbial suspensions . ImLA as a substrate remained without diminution in all microbial suspensions . Although ImAA was found to be degraded to a small extent (3.4-6.3%) only after 18 h incubation, neither His nor other metabolites were detected on the chromatograms . These results have been demonstrated for the first time in rumen microorganisms and suggest that His may be an essential amino acid for rumen microorganisms. Appl Microbiol Biotechnol, 2001 Mar, 55(2), 143 - 9 Plate screening methods for the detection of polysaccharase-producing microorganisms; Ruijssenaars HJ et al.; Polysaccharide-degrading enzymes (polysaccharases) are widely applied in industry . One of the sources of these enzymes are polysaccharide-degrading microorganisms . To obtain such microorganisms from enrichment cultures, strain collections or gene libraries, efficient plate screening methods are required that discriminate between intact and degraded polysaccharide . This can be achieved by making use of specific physicochemical properties of the polysaccharide, such as complex formation with dyes and gelling capacity, or by the application of dye-labelled polysaccharides . This review presents a survey of plate methods based on these principles . Both theoretical and practical aspects of the methods are discussed. Biosci Biotechnol Biochem, 2001 Mar, 65(3), 570 - 8 Investigation of various genotype characteristics for inosine accumulation in Escherichia coli W3110; Matsui H et al.; For the derivation of an inosine-overproducing strain from the wild type microorganism, it is known that the addition of an adenine requirement, removal of purine nucleoside hydrolyzing activity, removal of the feedback inhibition, and repression of key enzymes in the purine nucleotides biosynthetic pathway are essential . Thus, the disruption of purA (adenine requirement), deoD (removal of purine nucleosides phosphorylase activity), purR (derepression of the regulation of purine nucleotides biosynthetic pathway), and the insensitivity of the feedback inhibition of phosphoribosylpyrophosphate (PRPP) amidotransferase by adenosine 5'-monophosphate (AMP) and guanosine 5'-monophosphate (GMP) were done in the Escherichia coli strain W3110, and then the inosine productivity was estimated . In the case of using a plasmid harboring the PRPP amidotransferase gene (purF) that encoded a desensitized PRPP amidotransferase, purF disrupted mutants were used as the host strains . It was found that the innovation of the four genotypes brought about a small amount of inosine accumulation . Furthermore, an adenine auxotrophic mutant of E . coli showed inappropriate adenine use because its growth could not respond efficiently to the concentration of adenine added . As the presence of adenosine deaminase is well known in E . coli and it is thought to be involved in adenine use, a mutant disrupted adenosine deaminase gene (add) was constructed and tested . The mutant, which is deficient in purF, purA, deoD, purR, and add genes, and harboring the desensitized purF as a plasmid, accumulated about 1 g of inosine per liter . Although we investigated the effects of purR disruption and purF gene improvement, unexpectedly an increase in the inosine productivity could not be found with this mutant. Epidemiol Mikrobiol Imunol, 2001 Apr, 50(2), 87 - 91 {Free radicals in immunology and infectious diseases}; Racek J et al.; Free radicals contribute significantly in modification of immune processes and inflammatory reactions . They are produced by activated phagocytes which use them for killing microorganisms . Free radicals facilitate production of cytokines, which are important as modifiers of inflammatory reactions . Formation of free radicals is influenced by antioxidants which can thus modify the intensity of inflammatory reaction and immune response . The authors describe in detail the contribution of free radicals in etiology and pathogenesis of autoimmune diseases including rheumatoid arthritis, multiple sclerosis or amyotrophic lateral sclerosis . The role of free radicals and modifying influence of antioxidants in viral, bacterial, parasitic and mycotic diseases is described in the second part of the review . Finally, influence of free radicals and antioxidants on immunity changes in patients with malignant tumours, during aging and physical exercise is discussed. Xenotransplantation, 2001 May, 8(2), 125 - 35 Sensitive and specific immunological detection methods for porcine endogenous retroviruses applicable to experimental and clinical xenotransplantation; Tacke SJ et al.; The use of organs from transgenic pigs for xenotransplantation may be associated with the risk of transmission of microorganisms, especially when the transgenic pigs express human proteins influencing complement activation . The porcine endogenous retroviruses (PERVs) are of particular concern as they can infect human cells in vitro . However, it is unknown whether PERVs can infect transplant recipients in vivo and, if so, whether they are pathogenic . It is therefore essential for experimental and clinical xenotransplantation procedures that specific and sensitive screening methods for PERVs are established . We developed Western blot and enzyme-linked immunosorbant assays (ELISA) based on purified PERVs produced by pig and human cells or recombinant viral protein and synthetic peptides corresponding to PERVs' transmembrane envelope protein, respectively . PERV-specific anti-sera generated against purified virus particles, purified viral proteins and synthetic peptides served as positive controls . Both assays were used for screening the sera of healthy blood donors, pregnant women, patients treated with pig tissues, and butchers with extensive contact to living porcine material to detect antibodies against PERV . None of the individuals showed an antibody pattern characteristic for retroviral infections . Some individuals had antibodies reactive against the major capsid protein p27, against smaller viral proteins of the group specific antigen (Gag) in Western blot assays, or against peptides in the ELISA, probably due to cross-reactivity . Here, we present specific and highly sensitive screening methods applicable for future xenotransplantation procedures, but using these methods we found no evidence of PERV-infection among humans potentially at risk. Acta Neurol Scand, 2001 Apr, 103(4), 207 - 13 Respiratory complications related to bulbar dysfunction in motor neuron disease; Hadjikoutis S et al.; Bulbar dysfunction resulting from corticobulbar pathway or brainstem neuron degeneration is one of the most important clinical problems encountered in motor neuron disease (MND) and contributes to various respiratory complications which are major causes of morbidity and mortality . Chronic malnutrition as a consequence of bulbar muscle weakness may have a considerable bearing on respiratory muscle function and survival . Abnormalities of the control and strength of the laryngeal and pharyngeal muscles may cause upper airway obstruction increasing resistance to airflow . Bulbar muscle weakness prevents adequate peak cough flows to clear airway debris . Dysphagia can lead to aspiration of microorganisms, food and liquids and hence pneumonia . MND patients with bulbar involvement commonly display an abnormal respiratory pattern during swallow characterized by inspiration after swallow, prolonged swallow apnoea and multiple swallows per bolus . Volitional respiratory function tests such as forced vital capacity can be inaccurate in patients with bulbofacial weakness and/or impaired volitional respiratory control . Bulbar muscle weakness with abundant secretions may increase the risk of aspiration and make successful non-invasive assisted ventilation more difficult . We conclude that an evaluation of bulbar dysfunction is an essential element in the assessment of respiratory dysfunction in MND. IUBMB Life, 2000 Oct-Nov, 50(4-5), 267 - 9 The NADPH oxidase of endothelial cells; Babior BM; The best known NADPH oxidase is that of phagocytes-neutrophils and monocytes . In these cells, the enzyme manufactures large quantities of O2- and other reactive oxidants that are used for the purpose of killing invading microorganisms . Recent studies, however, have suggested that a number of other tissues contain NADPH oxidases . In contrast to the very vigorous production of oxidants by phagocytes, rates of oxidant production by these other cell types are quite low . Oxidant production by these cells is generally thought to serve a signaling function. J Rheumatol, 2001 Apr, 28(4), 802 - 4 Musculoskeletal manifestations in patients positive for human immunodeficiency virus: correlation with CD4 count; Casado E et al.; OBJECTIVE: To determine the relationship between the CD4+ lymphocyte count and musculoskeletal manifestations of human immunodeficiency virus (HIV) infection . METHODS: All patients from 1991 to 1998 who were positive for HIV with osteoarticular manifestations were reviewed retrospectively . HIV positivity was confirmed by ELISA and Western blot . CD4 count was performed by flow cytometry . RESULTS: We studied 74 patients with osteoarticular manifestations . The study group comprised 61 men (82.4%) and 13 women (17.5%) with a mean age of 34.2 years (range 17-62) . Fifty-two patients were iv drug users (70.3%) . Septic arthritis was present in 20 cases (23.0%), soft tissue infections in 9 cases (12.2%), spondyloarthropathies in 6 cases (8.1%), lymphomas in 9 cases (12.2%), osteomyelitis in 6 (8.1%), and 24 miscellaneous cases (32.4%) . The mean CD4 count was as follows: septic arthritis 164.7 cells/mm3, soft tissue involvement 127.1 cells/mm3, spondyloarthropathies 245.8 cells/mm3, lymphoma 132.8 cells/mm3, and osteomyelitis 233.6 cells/mm3 . CONCLUSION: Osteoarticular manifestations in the setting of HIV infection tend to be predominantly infectious . S . aureus is the microorganism most frequently involved . Ostearticular infections always appeared when the CD4 count was < 200 and pyomyositis and lymphoma appeared when CD4 was < 150 . CD4 counts may be useful predictors to determine the type of musculoskeletal manifestation. J Calif Dent Assoc, 2000 Mar, 28(3), 204 - 14 Oral microorganisms and cardiovascular disease; Pallasch TJ et al.; The list of etiological factors for cardiovascular disease is long, complicated, intertwined, and yet to be completed . This paper will evaluate the current evidence for the pathogenic role of certain microorganisms, including those of the oral cavity, in the etiology of cardiovascular disease. J Calif Dent Assoc, 2000 Mar, 28(3), 194 - 200 The focal infection theory: appraisal and reappraisal; Pallasch TJ et al.; This paper discusses the past, present, and future of the focal infection theory of disease . A focal infection is a localized or general infection caused by the dissemination of microorganisms or toxic products from a focus of infection . The resurgence of the focal infection theory of disease has been greeted with great enthusiasm in some quarters; however, the present evidence for the relationship of oral microorganisms and systemic disease is very limited due not only to a dearth of prospective studies and a complete lack of interventional studies but also to very significant methodological difficulties associated with the clinical studies that have been performed. Mol Psychiatry, 2001 May, 6(3), 329 - 33 Low avidity of human serum antibodies for Borna disease virus antigens questions their diagnostic value; Allmang U et al.; Borna disease virus (BDV) can induce neurological disease in animals . Since viral nucleic acid, infectious particles and antibodies recognizing BDV antigens were found at higher frequencies in psychiatric patients than in healthy controls, BDV is suspected to cause psychiatric disorders in humans . However, the human origin of these viruses has recently been questioned . To diagnose BDV infections, sera are usually analyzed for antiviral antibodies by indirect immunofluorescence (IFA) on virus-infected cells . This study reveals that the reactive antibodies in human sera mainly recognized the BDV phosphoprotein, whereas animal sera preferentially detected the viral nucleoprotein . Immunoglobulin (Ig) G in sera of experimentally or naturally infected animals bound to the viral antigen with high avidity, ie resisting 3 M urea, whereas reactive IgG in human sera did not . Longitudinal studies showed that reactive human antibodies persisted for many years without gaining high avidity for BDV antigens, indicating that they were probably not induced by BDV but rather by infection with an antigenically related microorganism of unknown identity or by exposure to other related immunogens. Nature, 2001 Apr 26, 410(6832), 1103 - 7 TREM-1 amplifies inflammation and is a crucial mediator of septic shock; Bouchon A et al.; Host innate responses to bacterial infections are primarily mediated by neutrophils and monocytes/macrophages . These cells express pattern recognition receptors (PRRs) that bind conserved molecular structures shared by groups of microorganisms . Stimulation of PRR signalling pathways initiates secretion of proinflammatory mediators, which promote the elimination of infectious agents and the induction of tissue repair . Excessive inflammation owing to bacterial infections can lead to tissue damage and septic shock . Here we show that inflammatory responses to microbial products are amplified by a pathway mediated by triggering receptor expressed on myeloid cells (TREM)-1 . TREM-1 is an activating receptor expressed at high levels on neutrophils and monocytes that infiltrate human tissues infected with bacteria . Furthermore, it is upregulated on peritoneal neutrophils of patients with microbial sepsis and mice with experimental lipopolysaccaride (LPS)-induced shock . Notably, blockade of TREM-1 protects mice against LPS-induced shock, as well as microbial sepsis caused by live Escherichia coli or caecal ligation and puncture . These results demonstrate a critical function of TREM-1 in acute inflammatory responses to bacteria and implicate TREM-1 as a potential therapeutic target for septic shock. J Mol Microbiol Biotechnol, 2001 Apr, 3(2), 145 - 50 Comparative genomics of microbial drug efflux systems; Paulsen IT et al.; The complete genome sequences of 36 microorganisms have now been published and this wealth of genome data has enabled the development of comparative genomic and functional genomic approaches to investigate the biology of these organisms . Comparative genomic analyses of membrane transport systems have revealed that transporter substrate specificities correlate with an organism's lifestyle . The types and numbers of predicted drug efflux systems vary dramatically amongst sequenced organisms . Microarray and gene knockout studies to date have suggested that predicted drug efflux genes often appear to be a) non-essential and b) expressed at detectable levels under standard laboratory growth conditions. Environ Microbiol, 2001 Feb, 3(2), 81 - 91 Chemical properties of catechols and their molecular modes of toxic action in cells, from microorganisms to mammals; Schweigert N et al.; Catechols can undergo a variety of chemical reactions . In this review, we particularly focus on complex formations and the redox chemistry of catechols, which play an inportant role in the toxicity of catechols . In the presence of heavy metals, such as iron or copper, stable complexes can be formed . In the presence of oxidizing agents, catechols can be oxidized to semiquinone radicals and in a next step to o-benzoquinones . Heavy metals may catalyse redox reactions in which catechols are involved . Further chemical properties like the acidity constant and the lipophilicity of different catechols are shortly described as well . As a consequence of the chemical properties and the chemical reactions of catechols, many different reactions can occur with biomolecules such as DNA, proteins and membranes, ultimately leading to non-repairable damage . Reactions with nucleic acids such as adduct formation and strand breaks are discussed among others . Interactions with proteins causing protein and enzyme inactivation are described . The membrane-catechol interactions discussed here are lipid peroxidation and uncoupling . The deleterious effect of the interactions between catechols and the different biomolecules is discussed in the context of the observed toxicities, caused by catechols. Environ Microbiol, 2001 Feb, 3(2), 145 - 8 BMRI-2, Rossendorf/Dresden, Germany (30 August - 1 September 2000); Hobman JL; Clearly, there is much left to be understood about microbial processes and interactions with metals, but much progress has been made, and the multidisciplinary approach of groups who are studying both the microbial populations and the chemistry of biotransformations of metals by bacteria will ensure rapid progress in our understanding of these issues . Several major points from different speakers summarize this meeting and are usefully reiterated at this point: Toxic metal ions, unlike organic pollutants, are immutable, and their bioavailability is a critical feature of their toxicity . The mobility, transport and fate of toxic metals and radionuclides in the environment are dependent on chemical and geochemical processes in which micro-organisms are intimately involved . Metals can be mobilized as well as immobilized by microorganisms . Metal/radionuclide valencies and chemical properties are critical to their environmental mobility . Bacterial- or fungal-metal interactions will be complicated by the presence of other pollutants . The identification of bacteria from environmental samples should not rely on one methodology, as these have been shown to be biased . Sonja Selenska-Pobell organized both BMRI-1 in 1998 and BMRI-2, which had well over 100 participants from Europe, Russia, USA and Japan in attendance . Thirty-one oral presentations were given, and over 30 posters were displayed over two poster sessions . BMRI-3 is provisionally planned for 2002 at GBF, Braunschweig, Germany. Clin Hemorheol Microcirc, 2000, 23(2-4), 345 - 8 In vivo identification of parasinus macrophages in the mesenteric lymph node; Nagata H et al.; Macrophages beneath the marginal sinus in the lymph nodes may play a role in defense against microorganism . The purpose of this study was to directly visualize the parasinus macrophages in the mesenteric lymph node . Fluorescent latex particles were injected into the appendix submucosa of rats . The mesenteric lymph node was epi-illuminated and observed with a fluorescent microscope . Fluorescent particles entered the marginal sinus of the mesenteric lymph node through the afferent lymphatic vessels, and distributed diffusely all over the marginal sinus . The particles became aggregated and interspersed 3 hr after injection, suggesting that particles were incorporated by phagocytes . The number of these particle-laden phagocytes increased up to 12 hr after injection, and then declined . Some phagocytes migrated rapidly within the marginal sinus . Morphology of these phagocytes in cell suspension was consistent with macrophages . In conclusion, we successfully visualized parasinus macrophages in vivo, which incorporated foreign bodies and migrated within the marginal sinus. Med Pregl, 2000 Nov-Dec, 53(11-12), 600 - 2 {Effect of alcohol consumption on recurrence of venereal diseases}; Bjekic M et al.; INTRODUCTION: Sexually transmitted diseases (STDs) comprise a large group of infections caused by different microorganisms including spirochetes, bacteria, chlamydia, mycoplasma, protozoa, fungi, parasites, and viruses . A considerable number of patients with sexually transmitted diseases are STD recurrences . As reported by Marijanovic and Lalosevic, in Belgrade, among patients who visited the City Department of Skin and Venereal Diseases, during 1985 and 1986 because of syphilis or gonorrhea, 22.8% had these diseases two or more times during their lives (male/female ratio 10:1) . The aim of this study was to test the hypothesis that there is a relationship between alcohol use and recurrence of STD . MATERIAL AND METHODS: A case-control study was performed in Belgrade population, from June, 1997 to April, 1998 . Participants were recruited among patients attending the City Department of Skin and venereal Diseases of Belgrade because of sexually transmitted diseases (syphilis, gonorrhea, nongonoccocal urethritis and genital warts) . The case group comprised 101 patients who already had STD two or more times in their personal histories . The control group consisted of 210 patients treated at the same institution for micotic diseases, patients who either never had STD or had it only once (13% of controls) in their personal histories . All participants were men aged 20 to 50 years and all were from Belgrade . Data on demographic characteristics, sexual history and sexual behavior, and data on antisocial behavior were collected from all participants using an anonymous questionnaire . In the present paper only data on alcohol use are presented . In the analysis of data chi 2-test was used . RESULTS: STD recurrence patients in comparison to their controls used alcohol more frequently (56.3%:16.1%), especially hard liquors, and 55.5% of them used alcohol at the time of STD infection . DISCUSSION: In the present study STD recurrence patients consumed alcohol more frequently than their controls, especially hard liquors . In the study of Myliueva et al, 50% of venereal disease patients consumed alcohol now and then and 10% consumed alcohol frequently . Scheidt and Windle found that 60% of alcoholics had at least one sexually transmitted disease as the result of a high number of sexual partners, low use of condoms and practicing sex for drugs or money . Alcohol has a depressive effect on central nervous system, reduces anxiety and increases libido . CONCLUSION: The obtained results support the hypothesis that alcohol use is related to recurrence of STDs. Cancer Chemother Pharmacol, 2001 Mar, 47(3), 277 - 9 Typhlitis (neutropenic enterocolitis) after a single dose of vinorelbine; Ferrazzi E et al.; We report a case of a 50-year-old man with pretreated adenocarcinoma of the lung, who developed fatal neutropenic enterocolitis (typhlitis) after a standard dose of the cytotoxic drug vinorelbine . Blood cultures were negative for all microorganisms tested for . Stool cultures were negative for enteric rods but direct examination of fresh stool revealed the presence of Giardia lamblia . Abdominal pain and diarrhoea developed very rapidly while the patient was only moderately neutropenic . Metronidazole was prescribed without clinical benefit: the abdominal pain remained stable . The duration of neutropenia was very short (4 days) . The abdominal catastrophe ending in shock occurred after complete recovery of the neutrophil count . Neutropenic colitis has been reported with increasing frequency in solid tumours after the introduction of taxanes . This complication has been observed mainly in phase I studies, near the maximally tolerated doses (MTD) . The combined use of vinorelbine has recently been reported to exacerbate the toxic effects of taxane on the colon . The case presented here demonstrates that typhlitis can occur even with vinorelbine alone, used at a standard recommended dose (30 mg/m2). J Ind Microbiol Biotechnol, 2000 Dec, 25(6), 281 - 287 Human pathogens associated with raw produce and unpasteurized juices, and difficulties in decontamination; Burnett SL et al.; The ability of public health agencies to identify, through enhanced epidemiologic and surveillance techniques, raw fruits, vegetables, and unpasteurized juices as probable sources of infectious microorganisms, has undoubtedly resulted in increased numbers of documented outbreaks . Changes in agronomic, harvesting, distribution, processing, and consumption patterns and practices have also likely contributed to this increase . The risk of illness associated with raw produce and unpasteurized produce products can be reduced by controlling or preventing contamination, or by removing or killing pathogenic microorganisms by washing or treating them with sanitizers . However, the hydrophobic cutin, diverse surface morphologies, and abrasions in the epidermis of fruits and vegetables limit the efficacy of these treatments. Acta Crystallogr D Biol Crystallogr, 2001 May, 57(Pt 5), 737 - 9 Epub 2001 Apr 24. Purification, crystallization and preliminary X-ray analysis of two crystal forms of ribonuclease Sa3; Hlinkova V et al.; RNase Sa3 produced by Streptomyces aureofaciens strain CCM 3239 belongs to the T1 family of microbial ribonucleases . It is closely related both to RNase Sa, studied in detail earlier, and to RNase Sa2 produced by the same microorganism . The most important property of RNase Sa3 is the relatively high cytotoxic activity, which was not observed for RNase Sa and Sa2 . Recombinant RNase Sa3 was overexpressed in Escherichia coli and purified to high homogeneity . The hanging-drop vapour-diffusion method was used for crystallization . The two crystal forms are trigonal P3(1)21 and tetragonal P4(1)2(1)2, with unit-cell parameters a = b = 64.7, c = 69.6 A, gamma = 120 degrees and a = b = 34.0, c = 147.2 A, respectively . They diffract to 2.0 and to 1.7 A resolution, respectively, using synchrotron radiation . The asymmetric units of crystal forms I and II contain one molecule of the enzyme, which corresponds to V(M) = 3.8 A(3) Da(-1) with a solvent content of 68% and V(M) = 1.9 A(3) Da(-1) with a solvent content of 37%, respectively. Acta Crystallogr D Biol Crystallogr, 2001 May, 57(Pt 5), 734 - 6 Epub 2001 Apr 24. Crystallization and preliminary X-ray crystallographic analysis of FlhD/FlhC complex from Escherichia coli; Wang S et al.; The heterotetrameric (C(2)D(2)) FlhD/FlhC complex was first discovered as a transcriptional activator of the flagellar genes in Escherichia coli . Recent studies now show that FlhD/FlhC also regulates several non-flagellar target genes in E . coli . The FlhD/FlhC complex also plays several important roles in other microorganisms . The molecular interactions between FlhD and FlhC, as well as the mechanisms by which the complex may vary its DNA-binding specificity, are not clear . Determination of the FlhD/FlhC crystal structure will provide insight into these protein-protein and protein-DNA interactions . The initial steps in this investigation are reported here: the overexpression, purification and crystallization of the FlhD/FlhC complex, the characterization of this crystal form and the recording and processing of an initial diffraction data set . The obtained crystal form of the FlhD/FlhC complex is hexagonal (space group P6(1), unit-cell parameters a = b = 150.5, c = 115.9 A) . The crystal density is very low (V(M) = 5.5), with 81.7% of its volume occupied by solvent . A single C(2)D(2) tetramer is present in the crystallographic asymmetric unit . A complete native data set has been collected to 4.5 A resolution. Appl Environ Microbiol, 2001 May, 67(5), 2336 - 44 Natural communities of novel archaea and bacteria growing in cold sulfurous springs with a string-of-pearls-like morphology; Rudolph C et al.; We report the identification of novel archaea living in close association with bacteria in the cold (approximately 10 degrees C) sulfurous marsh water of the Sippenauer Moor near Regensburg, Bavaria, Germany . These microorganisms form a characteristic, macroscopically visible structure, morphologically comparable to a string of pearls . Tiny, whitish globules (the pearls; diameter, about 0.5 to 3.0 mm) are connected to each other by thin, white-colored threads . Fluorescent in situ hybridization (FISH) studies have revealed that the outer part of the pearls is mainly composed of bacteria, with a filamentous bacterium predominating . Internally, archaeal cocci are the predominant microorganisms, with up to 10(7) cells estimated to be present in a single pearl . The archaea appear to be embedded in a polymer of unknown chemical composition . According to FISH and 16S rRNA gene sequence analysis, the archaea are affiliated with the euryarchaeal kingdom . The new euryarchaeal sequence represents a deep phylogenetic branch within the 16S rRNA tree and does not show extensive similarity to any cultivated archaea or to 16S rRNA gene sequences from environmental samples. Am J Respir Crit Care Med, 2001 Apr, 163(5), 1198 - 205 Attenuation of interleukin 8-induced nasal inflammation by an inhibitor peptide; Cooper JA Jr et al.; Polymorphonuclear neutrophils (PMNs) infiltrate tissue in response to chemoattractants, including interleukin 8 (IL-8) . Infiltrating PMNs clear microorganisms but also cause tissue damage . We previously reported the presence in human bronchial lavage of a peptide that inhibits PMN functions . The current project assessed (1) effects of a synthetic analog of this peptide (synthetic neutrophil inhibitor peptide, SNIP) on IL-8-induced nasal inflammation in humans, (2) effects of SNIP on PMN apoptosis and chemotaxis, (3) specific binding of SNIP to PMNs, and (4) evidence of larger molecules with the SNIP sequence . Results show that SNIP attenuates IL-8-induced nasal inflammation, inhibits in vitro PMN chemotaxis to IL-8, and accentuates PMNs apoptosis . PMNs contain specific SNIP-binding sites and the integrin CR3 (CD11b/CD18), or a CR3-associated molecule, is one SNIP-binding molecule . Chemotaxis to IL-8 is most potently inhibited by SNIP in the presence of fibrinogen, a CR3 ligand . Antiserum against the SNIP sequence recognizes a 70-kDa protein in bronchoalveolar lavage and an anti-SNIP immunoaffinity column binds a 70-kDa protein in U937 cell culture supernatant . U937 cell mRNA contains a 1.8-kb transcript detected with degenerate oligonucleotides designed from the SNIP sequence . These studies demonstrate that a synthetic inhibitor peptide can attenuate in vivo nasal inflammation through downregulatory effects on PMNs. Am J Respir Crit Care Med, 2001 Apr, 163(5), 1097 - 100 Increased frequency of Chlamydia pneumoniae antibodies in patients with asthma; Gencay M et al.; The worldwide increase in asthma incidences and the impact of the disease on public health care have led to new investigations of the cause of the disease . Besides well-defined environmental causes, accumulating evidence suggests that respiratory tract infections play an important role in the pathogenesis of asthma . Among these microorganisms Chlamydia pneumoniae is an intracellular pathogen causing persistent infection . Chlamydia pneumoniae infection has been discussed as possibly inducing the development of asthma . This study was designed to investigate the presence of C . pneumoniae-specific IgG, IgA, and IgM antibodies in serum samples of 33 adults with a clinical history of asthma, positive methacholine test, and reduced FEV(1) . Patients with asthma were compared with age-, sex-, and locality-matched control subjects (n = 33) . We observed no acute infection either in patients with asthma or in control subjects, but 63% of all investigated individuals had signs of past infection . Chlamydia pneumoniae-specific IgA was detected in 52% of the patients with asthma and in 15% of the healthy control subjects (p < 0.01) . Serological evidence of chronic infection with C . pneumoniae (high IgG {> pr = 1:512} and high IgA {> or = 1:40}) was more frequent in patients with asthma (18.2%) compared with control subjects (3.0%) (p < 0.01) . Our results provide further evidence that chronic infection with C . pneumoniae is linked to asthma. Res Microbiol, 2001 Mar, 152(2), 123 - 9 Genetic interference in protozoa; Bastin P et al.; RNA interference first described in Caenorhabditis elegans and transgene-induced post-transcriptional gene silencing first described in plants and fungi now appear as different means of activating a conserved and ancient mechanism that can protect genomes against viruses and transposons and perhaps also control expression of endogenous genes . We present here similar genetic interference phenomena in highly divergent protozoa, Trypanosoma and Paramecium, and look ahead to what contribution these microorganisms could bring to this fast-moving area. Mikrobiologiia, 2000 Sep-Oct, 69(5), 727 - 36 {Regulation of the biomass and activity of soil microorganisms by microfauna}; Mamilov ASh et al.; Microcosm experiments showed that the microbial biomass and the respiration activity in soil were regulated by nematodes . Depending on nematode number and plant residue composition, the trophic activity of nematodes can either stimulate or inhibit microbial growth and respiration as compared to soil containing no nematodes . The stimulating effect was observed when nitrogen-free (starch) or low-nitrogen (wheat straw, C:N = 87) organic substrates were applied . Inhibition occurred when a substrate rich in nitrogen (alfalfa meal, C:N = 28) was decomposed and the nematode population exceeded the naturally occurring level . A conceptual model was developed to describe trophic regulation by microfauna (nematodes) of the microbial productivity and respiration activity and decomposition of not readily decomposable organic matter in soil . The stimulating and inhibiting influence of microfauna on soil microorganisms was not a linear function of the rate of microbial consumption by nematodes . These effects are largely associated with the induced change in the physiological state of microorganisms rather than with the mobilization of biogenic elements from the decomposed microbial biomass. Mikrobiologiia, 2000 Sep-Oct, 69(5), 597 - 605 {Distant informational interactions in bacteria}; Nikolaev IuA; Exchange of information between bacteria via physical signals, referred to as "distant interactions" (DI), is the subject of this review . All cases of DI reported to date are discussed, as well as the history of these studies and the place of DI in bacterial communication . Bacterial DI are a particular case of DI occurring in nature (in plants, animals, and fungi) . Along with the chemical signals of intracellular communications, DI play a significant role in the life of microorganisms, especially during critical periods. Mem Inst Oswaldo Cruz, 2001 Apr, 96(3), 415 - 25 Coevolutionary networks: a novel approach to understanding the relationships of humans with the infectious agents; Tosta CE; Human organism is interpenetrated by the world of microorganisms, from the conception until the death . This interpenetration involves different levels of interactions between the partners including trophic exchanges, bi-directional cell signaling and gene activation, besides genetic and epigenetic phenomena, and tends towards mutual adaptation and coevolution . Since these processes are critical for the survival of individuals and species, they rely on the existence of a complex organization of adaptive systems aiming at two apparently conflicting purposes: the maintenance of the internal coherence of each partner, and a mutually advantageous coexistence and progressive adaptation between them . Humans possess three adaptive systems: the nervous, the endocrine and the immune system, each internally organized into subsystems functionally connected by intraconnections, to maintain the internal coherence of the system . The three adaptive systems aim at the maintenance of the internal coherence of the organism and are functionally linked by interconnections, in such way that what happens to one is immediately sensed by the others . The different communities of infectious agents that live within the organism are also organized into functional networks . The members of each community are linked by intraconnections, represented by the mutual trophic, metabolic and other influences, while the different infectious communities affect each other through interconnections . Furthermore, by means of its adaptive systems, the organism influences and is influenced by the microbial communities through the existence of transconnections . It is proposed that these highly complex and dynamic networks, involving gene exchange and epigenetic phenomena, represent major coevolutionary forces for humans and microorganisms. J Biol Chem, 2001 Jun 29, 276(26), 23456 - 63 Epub 2001 Apr 19. Human intelectin is a novel soluble lectin that recognizes galactofuranose in carbohydrate chains of bacterial cell wall; Tsuji S et al.; Galactofuranosyl residues are present in various microorganisms but not in mammals . In this study, we identified a human lectin binding to galactofuranosyl residues and named this protein human intelectin (hIntL) . The mature hIntL was a secretory glycoprotein consisting of 295 amino acids and N-linked oligosaccharides, and its basic structural unit was a 120-kDa homotrimer in which 40-kDa polypeptides were bridged by disulfide bonds . The hIntL gene was split into 8 exons on chromosome 1q21.3, and hIntL mRNA was expressed in the heart, small intestine, colon, and thymus . hIntL showed high levels of homology with mouse intelectin, Xenopus laevis cortical granule lectin/oocyte lectin, lamprey serum lectin, and ascidian galactose-specific lectin . These homologues commonly contained no carbohydrate recognition domain, which is a characteristic of C-type lectins, although some of them have been reported as Ca(2+)-dependent lectins . Recombinant hIntL revealed affinities to d-pentoses and a d-galactofuranosyl residue in the presence of Ca(2+), and recognized the bacterial arabinogalactan of Nocardia containing d-galactofuranosyl residues . These results suggested that hIntL is a new type lectin recognizing galactofuranose, and that hIntL plays a role in the recognition of bacteria-specific components in the host. Gastroenterology, 2001 May, 120(6), 1485 - 501 The lymphoid liver: considerations on pathways to autoimmune injury; Kita H et al.; Immunologic injury in the liver involves immigrant T and B lymphocytes and a resident lymphoid population that comprises distinct lymphocytic cells and accessory cells . The forerunner to autoimmunity is breaching of natural self-tolerance and hence the disruption of a fundamental property of the immune system . Such breaching occurs by processes that include inflammatory activation of immunocytes and macrophages, spillage of intracellular constituents, and epitope mimicry by constituents of microorganisms, with these acting on a genetically conditional phenotype; compounding factors include aberrations of apoptosis, whether insufficient or excess . The downstream end requires specifically directed inflammatory leukocyte traffic as an essential component of autoimmune expressions in the liver . The culmination is an orchestrated attack on hepatocytes or biliary epithelial cells by multiple effector pathways . Progress in type 1 autoimmune hepatitis still requires knowledge of a disease-specific autoantigen(s) involved in T-cell reactivity, although such knowledge in type 2 autoimmune hepatitis, in which the known autoantigen is cytochrome P4502D6, has not yet been integrated into a clearly defined scheme of pathogenesis . For PBC there has been a very promising amalgamation of molecular knowledge of the mitochondrial autoantigens . Future insights require deeper analysis of molecular, genetic, macroenvironmental, and microenvironmental elements in predisposition. Eur J Med Res, 2001 Jan 29, 6(1), 33 - 8 Surfactant proteins D and A in sputum; Griese M et al.; The surfactant proteins SP-D and SP-A are collectins and central components of the innate immune system of the peripheral lungs . They bind to carbohydrates on microorganisms and promote their removal . In the larger airways their role and concentrations are not yet known . Sputum may thus potentially be useful to characterize the collectins in this compartment . Induced sputum was obtained from healthy adults and spontaneous sputum from patients with cystic fibrosis . ELISA and carbohydrate binding assays were used to quantify the amount and functional capacity of the collectins in plugs or sol-fractions, prepared by centrifugation of the plugs . SP-D and SP-A were detectable in only about 50% of the sputum plugs . Varying the salt, detergent and calcium conditions of the assay did not improve the results . In samples with reproducibly detectable SP-D or SP-A, their carbohydrate binding capacity was zero . Sputum sol fraction and LPS inhibited the binding of the collectins to carbohydrate in the presence of calcium . Whereas SP-D and SP-A are abundant in the peripheral lung, their presence in sputum derived from the larger airways is variable and their carbohydrate binding capacity is lost. Biotechnol Prog, 2001 Mar-Apr, 17(2), 379 - 82 Enzymatic hydrolysis of waste office paper using viscosity as operating parameter; Park EY et al.; Enzymatic hydrolysis of waste office (WO) paper with feeding WO paper in a reactor was investigated using apparent viscosity as operating parameter . Since the apparent viscosity was correlated with the concentration of pulping WO paper, the amount of hydrolyzed WO paper was assumed by measuring the decrease in the apparent viscosity . Then the amount of hydrolysis WO paper and the amount of enzyme corresponding to the desired ratio were fed into the reactor . When the WO paper and 1% (to the amount of WO paper) enzyme were fed to the hydrolytic reaction, 87 g/L of reducing sugar (RS) with a hydrolytic yield of 42.2% was obtained for a 24-h hydrolysis . However, when nonpulping WO paper and 5% (to the amount of WO paper) enzyme were fed to the hydrolytic reaction, 120 g/L of RS with a hydrolytic yield of 40% was obtained for a 24-h hydrolysis . Therefore, the RS concentration from this hydrolysis process feeding WO paper using apparent viscosity as operating parameter may be of sufficient concentration to serve as a carbon source in microorganism culture or chemical feedstock. Luminescence, 2001 Mar-Apr, 16(2), 193 - 8 Applications of biochemiluminescence to HACCP; Champiat D et al.; The hazard analysis critical control point) (HACCP) system is set up with the aim of ensuring the harmlessness of food along the process chain, from the producer to the ultimate consumer . To set up this system, it is necessary to perform an analysis to identify the dangers and the critical points of the process in real time . Living organisms are detected on the principle of measurement of cellular energy in the form of adenosine triphosphate (ATP), which is produced by all living cells and of particularly by microorganisms . The principle of the bioluminescence of firefly luciferase is applied, whereby a reduced form of luciferin, together with a molecule of oxygen and ATP in the presence of the enzyme luciferase, produces a reaction in which oxyluciferine is released, together with AMP . This is accompanied by a simultaneous release of photons, which are then quantified . The emission spectra recorded with the firefly luciferin/luciferase system is obtained with absolute sensitivity . The method of ATP assays here proposed is adapted for measuring the total content of different adenylates in the cell, i.e . the adenylate pool . ATP concentration is then measured through bioluminescence, i.e . the light is measured by a photomultiplier tube detector and the signal is converted to relative light units (RLU) . Thus, RLU have a relationship with the amount of ATP present in the sample, which is not the case with traditional methods . Adequate levels of sensitivity are calculated with appropriate controls and the readings are translated into a statistical designation of positive or negative result . There are numerous illustrations of HACCP applications using biochemiluminescence (BCL) techniques as detection technology in food industry production lines and in control of critical points in real time in industrial applications . J Biol Chem, 2001 Jun 22, 276(25), 23056 - 64 Epub 2001 Apr 18. A spirochete surface protein uncouples store-operated calcium channels in fibroblasts: a novel cytotoxic mechanism; Wang Q et al.; The cytotoxicity of infectious agents can be mediated by disruption of calcium signaling in target cells . Outer membrane proteins of the spirochete Treponema denticola, a periodontal pathogen, inhibit agonist-induced Ca(2+) release from internal stores in gingival fibroblasts, but the mechanism is not defined . We determined here that the major surface protein (Msp) of T . denticola perturbs calcium signaling in human fibroblasts by uncoupling store-operated channels . Msp localized in complexes on the cell surface . Ratio fluorimetry showed that in cells loaded with fura-2 or fura-C18, Msp induced cytoplasmic and near-plasma membrane Ca(2+) transients, respectively . Increased conductance was confirmed by fluorescence quenching of fura-2-loaded cells with Mn(2+) after Msp treatment . Calcium entry was blocked with anti-Msp antibodies and inhibited by chelating external Ca(2+) with EGTA . Msp pretreatment reduced the amplitude of {Ca(2+)}(i) transients upon challenge with ATP or thapsigargin . In experiments using cells loaded with mag-fura-2 to report endoplasmic reticulum Ca(2+), Msp reduced Ca(2+) efflux from endoplasmic reticulum stores when ATP was used as an agonist . Msp alone did not induce Ca(2+) release from these stores . Msp inhibited store-operated influx of extracellular calcium following intracellular Ca(2+) depletion by thapsigargin and also promoted the assembly of subcortical actin filaments . This actin assembly was blocked by chelating intracellular Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester . The reduced amplitude of agonist-induced transients and inhibition of store-operated Ca(2+) entry due to Msp were reversed by latrunculin B, an inhibitor of actin filament assembly . Thus, Msp retards Ca(2+) release from endoplasmic reticulum stores, and it inhibits subsequent Ca(2+) influx by uncoupling store-operated channels . Actin filament rearrangement coincident with conformational uncoupling of store-operated calcium fluxes is a novel mechanism by which surface proteins and toxins of pathogenic microorganisms may damage host cells. Peptides, 2001 Apr, 22(4), 557 - 65 Recognition and oxidative metabolism of cyclodipeptides by hepatic cytochrome P450; Delaforge M et al.; Possible recognition of peptide derivatives by hepatic cytochrome P450 3A has been suggested by binding and metabolism of numerous pseudopeptidic compounds such as ergot derivatives and cyclosporin.Natural linear or cyclic dipeptides containing hydrophobic amino acids produced by microorganisms and present in mammals are able to interact with the P450 active site through either iron-amine interactions (Type II) or hydrophobic Type I interactions . P450 3A from dexamethasone-treated rats or yeast-expressed P450 human 3A4 are the most potent in such interactions, which are particularly strong with peptides containing a histidyl residue.Some cyclodipeptides are rapidly transformed by rat cytochrome P450 3A to mono- or dihydroxylated metabolites, with turnovers around 3 nmoles min(-1) P450(-1) . Linear peptides are poorly transformed in these conditions . This metabolism of cyclodipeptides occurs in 8 species including man.Such interactions and metabolism have only minor consequences in terms of P450 3A binding and metabolism of classical P450 3A substrates . These data reinforce the concept that, in addition to their effect on the regulation of P450 neosynthesis, naturally occurring endogenous peptides are also substrates of P450 3A . The physiological activities of these peptides may be modulated by their metabolism. Biosystems, 2001 Mar, 59(3), 159 - 83 Modelling of hexadecane degradation in continuous-flow cultures; Ebenhoh W et al.; Microorganisms of Wadden Sea sediments are able to degrade hydrocarbons in suspensions . (Berthe-Corti, L., Bruns, A., Hulsch, R., 1997 . J . Microb . Methods 29, 129-137) have observed in continuous culture experiments that the growth rate of microorganisms increases roughly proportional to the dilution rate . The growth rate is nearly independent of the oxygen saturation down to about 0.5% . Even at very low oxygen supply, corresponding to an oxygen saturation far below 0.1%, growth takes place at a reduced rate . In this paper, a model is presented which can reproduce the results of these experiments . The model treats the following processes, selection of the active fraction of microorganisms growing on hexadecane, uptake of hexadecane and transformation into palmitate as a first metabolic step, synthesis of biomass, respiration and exudation . The processes are regulated by the substrate concentration, the internal palmitate quota, the exudates' concentration and an inhibiting factor . For the experiments under very low oxygen conditions, the observed growth with reduced O(2)-consumption and CO(2)-production is modelled by assuming an anoxic metabolic pathway. Virus Res, 2001 May, 75(1), 69 - 73 Comparison of the sialic acid binding activity of transmissible gastroenteritis coronavirus and E . coli K99; Schwegmann C et al.; Transmissible gastroenteritis coronavirus (TGEV) and Escherichia coli K99 are both enteropathogenic for pigs with infections being most severe in neonate animals . For both microorganisms, a sialic acid binding activity has been shown to be an essential pathogenicity factor . Here we demonstrate with haemagglutination and haemagglutination-inhibition assays that TGEV and E . coli K99 differ in their sialic acid binding activities with respect to the type and amount of sialic acid residues required on the erythrocytes surface as well as with respect to the type of sialoglycoconjugate preferentially recognized . Intestinal mucins from piglets (12-14 days old) and adult animals were shown to inhibit TGEV to the same extent . From our results we conclude that E . coli K99 and TGEV interact with different sialoglycoconjugates to establish an intestinal infection . The implications for the enteropathogenicity of TGEV are discussed. J Microbiol Methods, 2001 Jun, 45(2), 77 - 87 An advanced molecular strategy to identify bacterial communities on art objects; Schabereiter-Gurtner C et al.; The application of culture-independent techniques based on molecular biological methods, especially on the PCR amplification of 16S rRNA genes, attempts to overcome some shortcomings of conventional cultivation methods and reveals far more complex bacterial communities on art objects than can be shown by cultivation methods . One of the major challenges of investigating microbial growth on art objects by molecular means is the extraction of DNA, due to small sample amounts and PCR inhibitors . In the present study, we introduce a DNA extraction protocol, which allowed the extraction of PCR-amplifiable DNA from samples derived from lime wall paintings and loamy soil underground . The DNA extracts were used to amplify 16S ribosomal fragments, which were subsequently analyzed by denaturing gradient gel electrophoresis (DGGE) . In parallel with the DGGE analysis, clone libraries containing PCR fragments of the ribosomal gene were constructed and clones were screened by DGGE . Clone libraries allow the inclusion of the entire 16S rDNA sequence in the phylogenetic analyses of microorganisms, providing a more reliable phylogenetic identification of microorganisms than is obtained from sequence analyses of excised and directly sequenced DGGE bands. Bioorg Med Chem, 2001 Mar, 9(3), 793 - 800 Biotransformation of a 4(20),11(12)-taxadiene derivative; Sun DA et al.; A 4(20),11(12)-taxadiene derivative was converted to hydroxylated derivatives by Cunninghamella elegans AS3.2033 and Cunninghamella elegans var chibaensis ATCC 20230 . Both microorganisms led to C-1 hydroxylations and conversion to a C-15-hydroxylated abeo-taxane . Additional products from the two fungi differed: a C-14 oxidation and a trans-cis isomerization of the cinnamoyl for one and an unprecedented hydroxylation at C-17 for the other. PDA J Pharm Sci Technol, 2001 Mar-Apr, 55(2), 65 - 86 Retention of water-borne bacteria by membrane filters . Part I: Bacterial challenge tests on 0.2 and 0.22 micron rated filters; Sundaram S et al.; The results of bacterial challenge tests conducted on several 0.2 and 0.22 micron rated "sterilizing grade" filter cartridge types with bacteria from a natural water source are presented . Eight different 0.2/0.22 micron rated "sterilizing grade" filter types from four different filter manufacturers, claimed to be capable of retaining Brevundimonas diminuta at a challenge level of 10(7) CFU/cm2, were tested . The filters tested included nylon 6.6 and polyamide filters from two manufacturers, modified or hydrophilic PVDF filters from two manufacturers, modified or asymmetric PES filters from three manufacturers, and cellulose acetate filters from a single manufacturer . Consistent bacterial penetration was observed, over the 18-24 h challenge period, for all twenty-five integral 0.2 and 0.22 micron rated filter cartridges tested, at challenge levels of about 10(1)-10(4) CFU/cm2, indicating that natural waterborne bacteria were more penetrative than B . diminuta . The observed penetration was thus qualitatively independent of filter media type or manufacturer . These results add to the growing body of evidence that shows 0.2 and 0.22 micron rated filters may not remove all microorganisms under all conditions . These results further establish that bacterial penetration of 0.2/0.22 micron rated filters is not limited just to (1) specific membrane types, or (2) extended duration challenges (>> 24 h), or (3) extremely high challenge levels, or (4) bacteria that can only exist in a penetrative state in an artificial laboratory setting. J Food Prot, 2001 Apr, 64(4), 451 - 5 Hot water immersion to eliminate Escherichia coli O157:H7 on the surface of whole apples: thermal effects and efficacy; Fleischman GJ et al.; The effect of hot water immersion on both the reduction of Escherichia coli O157:H7 on the apple surface and internal temperatures of the apple was assessed in this study . Microbial reductions were measured experimentally, whereas internal temperatures were calculated through a mathematical analysis of experimental heat transfer data obtained from the apples . A method was developed to provide a purely surface-based inoculation of E . coli O157:H7 . Rinsing produced no reduction, and treatments at 80 and 95 degrees C produced reductions of more than 5 logs in 15 s or less . The heat transfer analysis based on experimental data was used to calculate surface heat transfer coefficients and predict temperatures throughout the apple . The analysis indicated a low heat transfer rate . Although it reduces thermal degradation, a low heat transfer rate precludes thermal-based reduction of any internalized microorganisms. Int Endod J, 2000 Jan, 33(1), 1 - 18 Root canal treatment and general health: a review of the literature; Murray CA et al.; REVIEW: The focal infection theory was prominent in the medical literature during the early 1900s and curtailed the progress of endodontics . This theory proposed that microorganisms, or their toxins, arising from a focus of circumscribed infection within a tissue could disseminate systemically, resulting in the initiation or exacerbation of systemic illness or the damage of a distant tissue site . For example, during the focal infection era rheumatoid arthritis (RA) was identified as having a close relationship with dental health . The theory was eventually discredited because there was only anecdotal evidence to support its claims and few scientifically controlled studies . There has been a renewed interest in the influence that foci of infection within the oral tissues may have on general health . Some current research suggests a possible relationship between dental health and cardiovascular disease and published case reports have cited dental sources as causes for several systemic illnesses . Improved laboratory procedures employing sophisticated molecular biological techniques and enhanced culturing techniques have allowed researchers to confirm that bacteria recovered from the peripheral blood during root canal treatment originated in the root canal . It has been suggested that the bacteraemia, or the associated bacterial endotoxins, subsequent to root canal treatment, may cause potential systemic complications . Further research is required, however, using current sampling and laboratory methods from scientifically controlled population groups to determine if a significant relationship between general health and periradicular infection exists. Eur J Clin Microbiol Infect Dis, 2001 Feb, 20(2), 127 - 31 Specificity of a polymerase chain reaction assay of a target sequence on the 31-kilodalton Brucella antigen DNA used to diagnose human brucellosis; Casanas MC et al.; The aim of this study was to evaluate the specificity of a polymerase chain reaction assay for detecting Brucella DNA using primers specific for the amplification of a 223 bp region of the sequence encoding a 31 kDa immunogenic Brucella abortus protein (BCSP31) . DNA from all Brucella strains, including type, reference, vaccine and field strains, were correctly amplified . With the exception of Ochrobactrum spp., no other amplification was detected with a broad panel of microorganisms serologically or phylogenetically related to Brucella spp . This very good degree of specificity, together with its high yield demonstrated in previous clinical studies, confirms that this polymerase chain reaction assay could be a useful tool for the diagnosis of human brucellosis. J Clin Pathol, 2001 Apr, 54(4), 293 - 7 The physiological expression of inducible nitric oxide synthase (iNOS) in the human colon; Roberts PJ et al.; BACKGROUND: Inducible nitric oxide synthase (iNOS) is expressed in the colonic epithelium in both inflammatory bowel disease and colorectal cancer . Nitric oxide (NO), the product of this enzyme, has been implicated in the pathogenesis of both conditions . However, there are conflicting data on whether iNOS is expressed in the normal, uninflamed human colon . AIMS: To evaluate the expression of iNOS in histologically normal, non-inflamed human colonic mucosa . PATIENTS/METHODS: Reverse transcription polymerase chain reaction (RT-PCR), immunoblotting, and immunohistochemistry were used to investigate the expression of iNOS in 17 histologically normal specimens obtained at colectomy performed for colorectal neoplasia . In addition, 16 endoscopic mucosal biopsies, taken from normal individuals, were also evaluated . Eleven surgical specimens and 16 endoscopic biopsies from patients with refractory ulcerative colitis were used as inflammatory controls . RESULTS: All types of specimens expressed iNOS mRNA . Immunoblotting revealed a protein of approximately 130 kDa consistent with iNOS in mucosal extracts of 77% of normal individuals, and 85% of diseased controls . Immunolabelling localised this protein to the surface epithelium in most of the normal specimens and also to the crypt epithelium and inflammatory cells in the diseased controls . CONCLUSIONS: These findings provide evidence that iNOS is often expressed in the surface epithelium of non-inflamed human colon, suggesting that it is induced by local luminal factors, such as bacterial lipopolysaccharide (endotoxin) . The resultant NO produced at this site might act as an oxidative barrier, reducing bacterial translocation and providing a means of defence against pathogenic microorganisms. Biochem Biophys Res Commun, 2001 Apr 20, 282(5), 1237 - 43 Roles of histidine residues in tobacco acetolactate synthase; Oh KJ et al.; Acetolactate synthase (ALS) catalyzes the first common step in the biosynthesis of valine, leucine, and isoleucine in plants and microorganisms . ALS is the target of several structurally diverse classes of herbicides, including sulfonylureas, imidazolinones, and triazolopyrimidines . The roles of three well-conserved histidine residues (H351, H392, and H487) in tobacco ALS were determined using site-directed mutagenesis . Both H487F and H487L mutations abolished the enzymatic activity as well as the binding affinity for the cofactor FAD . Nevertheless, the mutation of H487F did not affect the secondary structure of the ALS . The K(m) values of H351M, H351Q, and H351F are approximately 18-, 60-, and fivefold higher than that of the wild-type ALS, respectively . Moreover, the K(c) value of H351Q for FAD is about 137-fold higher than that of wALS . Mutants H351M and H351Q showed very strong resistance to Londax (a sulfonylurea) and Cadre (an imidazolinone), whereas mutant H351F was weakly resistant to them . However, the secondary structures of mutants H351M and H351Q appeared to be different from that of wALS . The mutation of H392M