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| Int J Lepr Other Mycobact Dis, 2002 Jun, 70(2), 111 - 8 Contrary to BCG, MLM fails to induce the production of TNF alpha and NO by macrophages; Rojas-Espinosa O et al.; Pathogenic mycobacteria must possess efficient survival mechanisms to resist the harsh conditions of the intraphagosomal milieu . In this sense, Mycobacterium lepraemurium (MLM) is one of the most evolved intracellular parasites of murine macrophages; this microorganism has developed a series of properties that allows it not only to resist, but also to multiply within the inhospitable environment of the phagolysosome . Inside the macrophages, MLM appears surrounded by a thick lipid-envelope that protects the microorganism from the digestive effect of the phagosomal hydrolases and the acid pH . MLM produces a disease in which the loss of specific cell-mediated immunity ensues, thus preventing activation of macrophages . In vitro, and possibly also in vivo, MLM infects macrophages without triggering the oxidative (respiratory burst) response of these cells, thus preventing the production of the toxic reactive oxygen intermediaries (ROI) . Supporting the idea that MLM is within the most evolved pathogenic microorganisms, in the present study we found, that contrary to BCG, M . lepraemurium infects macrophages without stimulating these cells to produce meaningful levels of tumor necrosis factor alpha (TNF alpha) or nitric oxide (NO) . Thus, the ability of the microorganisms to stimulate in their cellular hosts, the production of ROI and RNI (reactive nitrogen intermediates), seems to be an inverse correlate of their pathogenicity; the lesser their ability, the greater their pathogenicity. Parasitology, 2002 Aug, 125(Pt 2), 177 - 85 Characterization of Meloidogyne javanica surface coat with antibodies and their effect on nematode behaviour; Sharon E et al.; The surface coat of the 2nd-stage juveniles (J2) of plant-parasitic nematodes is considered to be involved in interactions with microorganisms in the soil and rhizosphere, as well as with the host plant . Characterization of surface antigens might be important in the development of new nematode control strategies . In this study, polyclonal and monoclonal antibodies raised against Meloidogyne javanica, M . incognita and other plant-parasitic nematodes were tested for their binding to the surface coat and secreted-excreted products of M . javanica . Some of the monoclonal and polyclonal antibodies raised against M . incognita showed cross-reactivity with the surface coat of M . javanica . Western blot analysis of M . javanica surface coat extracts revealed labelling of several polypeptides with a 48 kDa main band for the polyclonal antibody IACR-PC Mi 373, and a 55 kDa main band for PC Mj E2 . Further characterization of the antigens recognized by the polyclonal antibody PC Mj E2, in planta, showed that they were present in the parasitic stages J2 and J3 and that the surface coat was shed during root penetration . The hypodermis of the infective juveniles was labelled by PC Mj E2 and the monoclonal antibody IACR-Misec 3F.4, suggesting that these surface antigens are produced in the hypodermis . Nematode behaviour was affected by all the antibodies that bound to the surface coat of the pre-parasitic J2, and we demonstrated that the movement pattern of the M . javanica J2 was affected by these antibodies . Continuous binding of the antibodies to the M . javanica surface inhibited the infection of Arabidopsis thaliana roots on agar plates. Proteins, 2002 Nov 1, 49(2), 216 - 31 Impact of incorporating the 2C5 crystal structure into comparative models of cytochrome P450 2D6; Kirton SB et al.; Cytochrome P450 2D6 (CYP2D6) metabolizes approximately one third of the drugs in current clinical use . To gain insight into its structure and function, we have produced four different sets of comparative models of 2D6: one based on the structures of P450s from four different microorganisms (P450 terp, P450 eryF, P450 cam, and P450 BM3), another on the only mammalian P450 (2C5) structure available, and the other two based on alternative amino acid sequence alignments of 2D6 with all five of these structures . Principal component analysis suggests that inclusion of the 2C5 crystal structure has a profound effect on the modeling process, altering the general topology of the active site, and that the models produced differ significantly from all of the templates . The four models of 2D6 were also used in conjunction with molecular docking to produce complexes with the substrates codeine and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP); this identified Glu 216 {in the F-helix; substrate recognition site (SRS) 2} as a key determinant in the binding of the basic moiety of the substrate . Our studies suggest that both Asp 301 and Glu 216 are required for metabolism of basic substrates . Furthermore, they suggest that Asp 301 (I-helix, SRS-4), a residue thought from mutagenesis studies to bind directly to the basic moiety of substrates, may play a key role in positioning the B'-C loop (SRS-1) and that the loss of activity on mutating Asp 301 may therefore be the result of an indirect effect (movement of the B'-C loop) on replacing this residue . Am J Phys Anthropol, 2002 Sep, 119(1), 27 - 36 Investigation of the link between visceral surface rib lesions and tuberculosis in a Medieval skeletal series from England using ancient DNA; Mays S et al.; Seven human skeletons from a large assemblage from a rural English Medieval burial site show lesions, predominantly proliferative in nature, on the visceral surfaces of the ribs . In order to investigate whether these rib lesions were regularly associated with tuberculous infection, these individuals, together with a group of age- and sex-matched control skeletons without bony signs of infection, were subjected to polymerase chain reaction (PCR) assays aimed at detecting traces of DNA from infecting microorganisms of the Mycobacterium tuberculosis complex . The results provided no evidence for any regular association between visceral surface rib lesions and the presence of M . tuberculosis complex DNA in the study group . The significance of these findings for the paleopathological interpretation of visceral surface rib lesions is discussed . Atherosclerosis, 2002 Nov, 165(1), 79 - 88 Chlamydia pneumoniae activates IKK/I kappa B-mediated signaling, which is inhibited by 4-HNE and following primary exposure; Donath B et al.; Chlamydia pneumoniae may be involved in atherosclerosis by inducing inflammation as well as LDL oxidation . The transcription factor NF-kappa B is found in an active state in atherosclerotic lesions . This study examined the effect of C . pneumoniae exposure on the NF-kappa B system in human monocytic lineage cells . Short exposure to C . pneumoniae as well as chlamydial heat shock protein 60 activated NF-kappa B, accompanied by increased cytokine production . Incubation with C . pneumoniae-induced depletion of I kappa B-alpha and later I kappa B-epsilon which was preceded by I kappa B kinase complex activation . 4-Hydroxynonenal, an aldehyde LDL oxidation product, was shown to inhibit C . pneumoniae induced NF-kappa B activation by preventing I kappa B phosphorylation/proteolysis . During long-term incubation with C . pneumoniae I kappa B-alpha returned to baseline, whereas the levels of I kappa B-epsilon and p65 were upregulated . Interestingly, long-term preincubation with C . pneumoniae selectively prevented restimulation by this microorganism, which appears to be at least partly facilitated by inhibition of I kappa B proteolysis . C . pneumoniae-induced NF-kappa B activation as well as the inhibition of that effect under certain conditions may contribute to chronic inflammation with potential relevance to vascular disease. Respir Res . 2002;3(1):21 . Epub 2002 Aug 07. Activation of adherent vascular neutrophils in the lung during acute endotoxemia; Sunil VR et al.; BACKGROUND: Neutrophils constitute the first line of defense against invading microorganisms . Whereas these cells readily undergo apoptosis under homeostatic conditions, their survival is prolonged during inflammatory reactions and they become biochemically and functionally activated . In the present study, we analyzed the effects of acute endotoxemia on the response of a unique subpopulation of neutrophils tightly adhered to the lung vasculature . METHODS: Rats were treated with 5 mg/kg lipopolysaccharide (i.v.) to induce acute endotoxemia . Adherent neutrophils were isolated from the lung vasculature by collagenase digestion and sequential filtering . Agarose gel electrophoresis, RT-PCR, western blotting and electrophoretic mobility shift assays were used to evaluate neutrophil activity . RESULTS: Adherent vascular neutrophils isolated from endotoxemic animals exhibited decreased apoptosis when compared to cells from control animals . This was associated with a marked increase in expression of the anti-apoptotic protein, Mcl-1 . Cells isolated 0.5-2 hours after endotoxin administration were more chemotactic than cells from control animals and expressed increased tumor necrosis factor-alpha and cyclooxygenase-2 mRNA and protein, demonstrating that they are functionally activated . Endotoxin treatment of the animals also induced p38 and p44/42 mitogen activated protein kinases in the adherent lung neutrophils, as well as nuclear binding activity of the transcription factors, NF-kappaB and cAMP response element binding protein . CONCLUSION: These data demonstrate that adherent vascular lung neutrophils are highly responsive to endotoxin and that pathways regulating apoptosis and cellular activation are upregulated in these cells. Science, 2002 Aug 30, 297(5586), 1543 - 5 Natural product terpenoids in Eocene and Miocene conifer fossils; Otto A et al.; Numerous saturated and aromatic hydrocarbons, but not polar compounds, originating from plants and microorganisms (biomarkers) have been reported in sediments, coals, and petroleum . Here we describe natural product terpenoids found in two fossil conifers, Taxodium balticum (Eocene) and Glyptostrobus oregonensis (Miocene) . A similar terpenoid pattern is also observed in extant Taxodium distichum . The preservation of characteristic terpenoids (unaltered natural products) in the fossil conifers supports their systematic assignment to the Cypress family (Cupressaceae sensu lato) . The results also show that fossil conifers can contain polar terpenoids, which are valuable markers for (paleo)chemosystematics and phylogeny. Water Sci Technol, 2002, 45(12), 175 - 9 Isolation and characterization of the ethynylestradiol-biodegrading microorganism Fusarium proliferatum strain HNS-1; Shi JH et al.; We cultivated hundreds of sediment, soil, and manure samples taken from rivers and farms in a medium containing ethynylestradiol (EE2) as the sole source of carbon, so that microorganisms in the samples would acclimatize to the presence of EE2 . Finally, we isolated an EE2-degrading microorganism, designated as strain HNS-1, from a cowshed sample . Based on its partial nucleotide sequence (563 bp) of the 28S rRNA gene, strain HNS-1 was identified as Fusarium proliferatum . Over 15 days, F . proliferatum strain HNS-1 removed 97% of EE2 at an initial concentration of 25 mg.L-1, with a first-order rate constant of 0.6 d-1 . Unknown products of EE2 degradation, which may be more polar compounds that have a phenolic group, remained in the culture medium. Appl Environ Microbiol, 2002 Sep, 68(9), 4431 - 40 Molecular evidence for a uniform microbial community in sponges from different oceans; Hentschel U et al.; Sponges (class Porifera) are evolutionarily ancient metazoans that populate the tropical oceans in great abundances but also occur in temperate regions and even in freshwater . Sponges contain large numbers of bacteria that are embedded within the animal matrix . The phylogeny of these bacteria and the evolutionary age of the interaction are virtually unknown . In order to provide insights into the species richness of the microbial community of sponges, we performed a comprehensive diversity survey based on 190 sponge-derived 16S ribosomal DNA (rDNA) sequences . The sponges Aplysina aerophoba and Theonella swinhoei were chosen for construction of the bacterial 16S rDNA library because they are taxonomically distantly related and they populate nonoverlapping geographic regions . In both sponges, a uniform microbial community was discovered whose phylogenetic signature is distinctly different from that of marine plankton or marine sediments . Altogether 14 monophyletic, sponge-specific sequence clusters were identified that belong to at least seven different bacterial divisions . By definition, the sequences of each cluster are more closely related to each other than to a sequence from nonsponge sources . These monophyletic clusters comprise 70% of all publicly available sponge-derived 16S rDNA sequences, reflecting the generality of the observed phenomenon . This shared microbial fraction represents the smallest common denominator of the sponges investigated in this study . Bacteria that are exclusively found in certain host species or that occur only transiently would have been missed . A picture emerges where sponges can be viewed as highly concentrated reservoirs of so far uncultured and elusive marine microorganisms. Acta Crystallogr D Biol Crystallogr, 2002 Sep, 58(Pt 9), 1494 - 6 Epub 2002 Aug 23. Crystallization and preliminary X-ray analysis of a xylanase from the psychrophile Pseudoalteromonas haloplanktis; Van Petegem F et al.; The 46 kDa xylanase from the Antarctic microorganism Pseudoalteromonas haloplanktis is an enzyme that efficiently catalyzes reactions at low temperatures . Here, the crystallization of both the native protein and the SeMet-substituted enzyme and data collection from both crystals using synchrotron radiation are described . The native data showed that the crystals diffract to 1.3 A resolution and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 50.87, b = 90.51, c = 97.23 A . SAD data collected at the peak of the selenium absorption edge proved to be sufficient to determine the heavy-atom configuration and to obtain electron density of good quality. Acta Crystallogr D Biol Crystallogr, 2002 Sep, 58(Pt 9), 1476 - 8 Epub 2002 Aug 23. Preliminary X-ray crystallographic analysis of a soluble form of MntC, a periplasmic manganese-binding component of an ABC-type Mn transporter from Synechocystis sp . PCC 6803; Adir N et al.; Manganese is recruited in microorganisms by way of ABC-type transporter systems . Here, the expression, purification and preliminary crystallographic analysis of a soluble form of the MntC solute-binding protein component of the MntABC manganese-import system from the cyanobacterium Synechococystis sp . PCC 6803 is reported . The protein (321 amino-acid residues) was expressed exclusively in inclusion bodies, which required unfolding and refolding in the presence of manganese prior to purification . The purified protein was crystallized in the presence of PEG and zinc . The crystals belong to space group P6(2)22, with unit-cell parameters a = b = 128.1, c = 90.0 A and a single molecule in the asymmetric unit . The crystals diffract to 2.6 A under cryoconditions using synchrotron radiation. Helicobacter, 2002, 7 Suppl 1, 50 - 5 Helicobacter pylori infection in pediatrics; Roma-Giannikou E et al.; In the past year the main interest was focused on the role of family for transmission of Helicobacter pylori to children; the evaluation of noninvasive diagnostic tests, especially in young children; extra-intestinal clinical manifestations; the lack of consensus on treatment; and the problem of high resistance of the microorganism to antibiotics. Vnitr Lek, 2002 Jul, 48(7), 657 - 66 {The role of infection and inflammation in the pathogenesis of atherosclerosis}; Jaber J et al.; Pathogenesis of the atherosclerotic process is deemed as multifactorial . To the most important risk factors, besides certain family predisposition, there belongs hypercholesterolemia, arterial hypertension, obesity, diabetes mellitus, smoking and others . In the last years there are more and more data about the role of inflammation and infection in the whole development of atherosclerosis . The witness for this hypothesis is the findings of high parameters of inflammation in involved vessels as well as in the blood of atherosclerosis suffering persons . Opinions about the inflammation theory appear from the 90th . Local sterile inflammation in the subendotelium of the middle and big arteries has been proved to consist of specific immune reaction (activation of the T-lymphocytes) as well as nonspecific characteristic by elevated monocytes in the artery wall during the whole process of atherogenesis . Inflammation in the plaque can trigger and hold several factors engaged in the atherosclerotic process, such as oxidized LDL cholesterol, elevated production of various superoxides, activated macrophages, activated T-lymphocytes, cytokines (IL-1, IL-6, interferon gamma) and lipoprotein Lp (a) . In this inflammation process levels of CRP (acute phase protein), fibrinogen and erythrocyte sedimentation are elevated as a reaction of the organism to nonspecific chronic infections . Because of this it is thought that elevated fibrinogen and erythrocyte sedimentation are markers of the cardiovascular risk . Some papers deal with antiinflammatory effects of statins, because these lower CRP levels so they also lower atherosclerotic risk through not only lowering of cholesterol levels . Also asprine, as an antiinflammation agent, changing the CRP levels, would be of benefit for patients with vascular disease because its antiaggregation and antiinflammatory effects . ACE inhibitors are also antiinflamatory through blocking of tissue production of angiotensin II (artery wall and atherosclerotic plaque) . Enzymatic inhibitors changing angiotensin can also have a partial antiinflammatory effect . The infection theory is supported also by tracing of some microorganisms in the atherosclerotic plaque or in the blood, as e.g . Helicobacter pylori or Chlamydia pneumoniae; to the autoimmune origin is indicated the presence of the specific immunity reaction against heat shock proteins (HSP) or oxidized LDL . This infection theory offers new therapy possibilities . Therefore eradication for example by antibiotics can lead to stabilization of the atherosclerotic plaque with positive consequences, as it was discovered by many studies. Diabetes Care, 2002 Sep, 25(9), 1534 - 8 Acidic drinking water and risk of childhood-onset type 1 diabetes; Stene LC et al.; OBJECTIVE: To estimate the associations of acidity and concentration of selected minerals in household tap water with the risk of type 1 diabetes . RESEARCH DESIGN AND METHODS: We designed a population-based case-control study with 64 cases of type 1 diabetes and 250 randomly selected control subjects . Acidity, color, and mineral content were measured in tap water from each participant's household . RESULTS: Tap water pH 6.2-6.9 was associated with a fourfold higher risk of type 1 diabetes compared with pH > or =7.7 (OR 3.73, 95% CI 1.52-9.15) . This result was similar after exclusion of individuals with the highly protective HLA-DQB1*0602 allele, but adjustment for maternal education, urban/rural residence, sex, and age tended to strengthen the estimated association . Higher tap water concentration of zinc was associated with lower risk of type 1 diabetes after adjustment for pH and other possible confounders, but the overall association was strictly not significant . CONCLUSIONS: These results suggest the possibility that quality of drinking water influences the risk of type 1 diabetes . The possible mechanisms by which water acidity or mineral content may be involved in the etiology of type 1 diabetes remain unknown, but the mechanisms are most likely indirect and may involve an influence on survival of microorganisms in the water. Genetics, 2002 Aug, 161(4), 1395 - 410 The tRNA-Tyr gene family of Saccharomyces cerevisiae: agents of phenotypic variation and position effects on mutation frequency; Ito-Harashima S et al.; Extensive phenotypic diversity or variation exists in clonal populations of microorganisms and is thought to play a role in adaptation to novel environments . This phenotypic variation or instability, which occurs by multiple mechanisms, may be a form of cellular differentiation and a stochastic means for modulating gene expression . This work dissects a case of phenotypic variation in a clinically derived Saccharomyces cerevisiae strain involving a cox15 ochre mutation, which acts as a reporter . The ochre mutation reverts to sense at a low frequency while tRNA-Tyr ochre suppressors (SUP-o) arise at a very high frequency to produce this phenotypic variation . The SUP-o mutations are highly pleiotropic . In addition, although all SUP-o mutations within the eight-member tRNA-Tyr gene family suppress the ochre mutation reporter, there are considerable phenotypic differences among the different SUP-o mutants . Finally, and of particular interest, there is a strong position effect on mutation frequency within the eight-member tRNA-Tyr gene family, with one locus, SUP6, mutating at a much higher than average frequency and two other loci, SUP2 and SUP8, mutating at much lower than average frequencies . Mechanisms for the position effect on mutation frequency are evaluated. Hamostaseologie, 2002 Feb, 22(1), 43 - 7 {Inhibitors of factor XIIIa}; Prasa D et al.; Factor XIIIa (FXIIIa) catalyzes the covalent crosslinking of fibrin polymers and incorporation of proteins into the fibrin network and thus confers on the thrombus additional structural stability and relative resistance to plasmin-mediated degradation . Moreover, FXIIIa is involved in other physiological and pathophysiological processes such as wound healing and arteriosclerosis . Selective FXIIIa inhibitors may be a valuable tool for evaluation of the various functions of FXIIIa and their pharmacological control . This paper presents an overview of the inhibitors of FXIIIa . Analogues of natural FXIIIa substrates - including glutamine containing peptides and low molecular weight substituted alkylamines - are incorporated into the fibrin network and thus prevent crosslinking of fibrin . Naturally occurring, direct inhibitors of FXIIIa have been isolated from a leech species and microorganisms . With effective concentrations in the nanomolar range the peptide tridegin is the most potent FXIIIa inhibitor up to now . The majority of the synthetic, low molecular Weight inhibitors bind covalently to Cys314 at the active site of FXIIIa . Besides the relatively nonspecific thiol reagents, azol derivatives, azolium salts and related substances are described as specific inhibitors of FXIIIa . They inhibit the activity of FXIIIa at nanomolar concentrations . Animal experiments have demonstrated improved thrombolysis by a plasminogen activator in combination with a FXIIIa inhibitor. J Immunol, 2002 Sep 1, 169(5), 2719 - 26 Functional activation of myelin-specific T cells by virus-induced molecular mimicry; Olson JK et al.; Molecular mimicry is the process by which T cells activated in response to determinants on an infecting microorganism cross-react with self epitopes, leading to an autoimmune disease . Normally, infection of SJL/J mice with the BeAn strain of Theiler's murine encephalomyelitis virus (TMEV) results in a persistent CNS infection, leading to a chronic progressive, CD4(+) T cell-mediated demyelinating disease . Myelin damage is initiated by T cell responses to virus persisting in CNS APCs, and progressive demyelinating disease (50 days postinfection) is perpetuated by myelin epitope-specific CD4(+) T cells activated by epitope spreading . We developed an infectious model of molecular mimicry by inserting a sequence encompassing the immunodominant myelin epitope, proteolipid protein (PLP) 139-151, into the coding region of a nonpathogenic TMEV variant . PLP139-TMEV-infected mice developed a rapid onset paralytic inflammatory, demyelinating disease paralleled by the activation of PLP139-151-specific CD4(+) Th1 responses within 10-14 days postinfection . The current studies demonstrate that the early onset demyelinating disease induced by PLP139-TMEV is the direct result of autoreactive PLP139-151-specific CD4(+) T cell responses . PLP139-151-specific CD4(+) T cells from PLP139-TMEV-infected mice transferred demyelinating disease to naive recipients and PLP139-151-specific tolerance before infection prevented clinical disease . Finally, infection with the mimic virus at sites peripheral to the CNS induced early demyelinating disease, suggesting that the PLP139-151-specific CD4(+) T cells could be activated in the periphery and traffic to the CNS . Collectively, infection with PLP139-151 mimic encoding TMEV serves as an excellent model for molecular mimicry by inducing pathologic myelin-specific CD4(+) T cells via a natural virus infection. Sheng Wu Gong Cheng Xue Bao, 2002 May, 18(3), 276 - 81 {Gene and gene engineering of carotenoid biosynthesis}; Tao J et al.; Carotenoids have a range of diverse biological functions and actions, especially playing an important role in human health with provitamin A activity, anti-cancer activity, enhancing immune ability and so on . Human body can't synthesis carotenoids by itself and must absorb them from outside . However, carotenoid contents in many plant are very low, and many kinds of carotenoid are difficult to produce by chemical ways . With the elucidation of carotenoid biosynthetic pathway and cloning genes of relative enzymes from microorganisms and higher plants, it is possible to regulate carotenoid biosynthesis via genetic engineering . This article reviews gene cloning of carotenoid biosynthetic enzymes in microorganisms and higher plants, and advances in the studies of carotenoid production in heterologous microorganisms and crop plants using gene-manipulated carotenoid biosynthesis. Immunol Rev, 2002 Jul, 185, 136 - 58 Cellular immunology in a historical perspective; Good RA; Bruton's XLA and DiGeorge syndrome patients show that two basic immune systems are distinct from each other in humans - thymus-dependent cell-mediated immunodeficiencies vs . antibody-based immunodeficiencies . The appendix-sacculus lymphoid organ of rabbits, like the bursa of Fabricius, represents a central lymphoid organ . Chronic granulomatous disease of childhood (CGD) revealed that phagocytosis killing of catalase-positive microorganisms employ oxidative burst . Bone marrow transplantation (BMT) proved life saving in severe combined immunodeficiency (SCID) . The first BMT cured XSCID and the second BMT cured a complicating aplastic anemia launching BMT as a treatment of many diseases . Now 75 fatal diseases have been cured by myeloablative BMT . BMT also cured experimental autoimmune diseases . BMT alone did not cure lupus with polyarthritis in MRL/lpr mice or polyarthritis in NZB/KN mice, but BMT plus bone (stromal cell) transplants cured these diseases . Autoimmune diseases and lethal glomerulonephritis were prevented or cured in BXSB mice by mixed allogeneic plus syngeneic BMT . X-linked Hyper IgM syndrome (XHIM) was also cured by BMT from a 2-year-old MHC-matched sibling donor . Nonmyeloablative BMT plus mesenchymal stem cells (stromal cells) was effective treatment for a form of collagen-vascular disease and also a lethal form of hypophosphatasia . Mannan-binding lectin, an opsonin that activates the complement system when mutated and at low levels in blood, opens a door to frequent infections throughout childhood and adult life . This new immunodeficiency is based on genetic mutations that involve a native defense system. South Med J, 2002 Aug, 95(8), 884 - 8 Incidental granulomatous inflammation of the uterus; Hoff E et al.; BACKGROUND: Granulomas of the uterine corpus have been reported in a variety of pathologic conditions but are relatively rare findings in routine histopathologic material . METHODS: This retrospective clinicopathologic study reviewed patients diagnosed with uterine granulomas between 1980 and 1999 in a tertiary referral center . RESULTS: The study group was comprised of 11 women, ranging in age from 37 to 90 years . All patients had histologically confirmed, non-necrotizing granulomas . The most common symptom prompting biopsy or hysterectomy was abnormal bleeding . Several concomittant histopathologies were noted . Eight of 11 patients had a known history of uterine instrumentation . None of the patients had clinical evidence of sarcoidosis or systemic infection, and stains for microorganisms were negative in all cases . Polarizable or foreign materials were not seen . CONCLUSIONS: Well-formed, non-necrotizing granulomas are an infrequent finding in the uterus . A history of instrumentation may explain the presence of granulomas in a subset of patients. Pol J Vet Sci, 2002, 5(2), 99 - 101 Pathogenicity of the microorganisms of the family Chlamydiaceae respecting the changes in their classification; Niemczuk K et al.; The comparison of names of the species before the reclassification of the family Chlamydiaceae into two genera with the species names acknowledged at present is presented . The pathogenicity of these species for animals and man is characterised. J Agric Food Chem, 2002 Aug 28, 50(18), 5167 - 78 Chitosan as an edible invisible film for quality preservation of herring and atlantic cod; Jeon YJ et al.; The effect of chitosan with different molecular weights as coatings for shelf-life extension of fresh fillets of Atlantic cod (Gadus morhua) and herring (Clupea harengus) was evaluated over a 12-day storage at refrigerated temperature (4 +/- 1 degrees C) . Three chitosan preparations from snow crab (Chinoecetes opilio) processing wastes, differing in viscosities and molecular weights, were prepared; their apparent viscosities (360, 57, and 14 cP) depended on the deacetylation time (4, 10, and 20 h, respectively) of the chitin precursor . Upon coating with chitosans, a significant (p < or = 0.05) reduction in relative moisture losses of 37, 29, 29, 40, and 32% was observed for cod samples coated with 360 cP chitosan after 4, 6, 8, 10, and 12 days of storage, respectively . Chitosan coating significantly (p < or = 0.05) reduced lipid oxidation as displayed in peroxide value, conjugated dienes, 2-thiobarbituric acid reactive substances and headspace volatiles, chemical spoilage as reflected in total volatile basic nitrogen, trimethylamine, and hypoxanthine, and growth of microorganisms as reflected in total plate count in both fish model systems compared to uncoated samples . The preservative efficacy and the viscosity of chitosan were inter-related; the efficacy of chitosans with viscosities of 57 and 360 cP was superior to that of chitosan with a 14 cP viscosity . Thus, chitosan as edible coating would enhance the quality of seafoods during storage. Water Sci Technol, 2002, 45(10), 313 - 9 Novel anaerobic process for the recovery of methane and compost from food waste; Han SK et al.; Multi-step sequential batch two-phase anaerobic composting (MUSTAC) process was used to recover methane and composted material from food waste . The MUSTAC process consists of five leaching beds for hydrolysis, acidification and post-treatment, and an upflow anaerobic sludge blanket (UASB) reactor for methane recovery . This process involves the combined methods of sequential batch operation and two-phase anaerobic digestion for simple operation and high efficiency . Rumen microorganisms are inoculated due to their enhanced cellulolytic activity . Each leaching bed is operated in a sequential batch mode . Five leaching beds are operated in a multi-step mode with a two-day interval between degradation stages . Acidified products in the leachate from the leaching beds are converted to methane in the UASB reactor . The MUSTAC process demonstrated that it was capable of removing 84.9% of volatile solids (VS) and converting 85.6% of biochemical methane potential (BMP) into methane at 10.9 kg VS/m3 x d in 10 days . Methane gas production rate was 2.31 m3/m3 x d . The output from the post-treatment of residues in the same leaching bed without troublesome moving met the Korean regulation on compost, indicating that it could be used for soil amendment. Environ Sci Technol, 2002 Aug 1, 36(15), 3290 - 4 Dechlorination of chlorobenzenes by a culture containing bacterium DF-1, a PCB dechlorinating microorganism; Wu Q et al.; Polychlorinated benzenes were reductively dechlorinated by an enrichment culture containing the polychlorinated biphenyl (PCB) dechlorinating bacterium DF-1 . The culture dechlorinated hexachlorobenzene (hexa-CB) --> pentachlorobenzene (penta-CB) --> 1,2,3,5-tetrachlorobenzene (1,2,3,5-CB) --> 1,3,5-trichlorobenzene (1,3,5-CB) and did not dechlorinate other tetrachlorobenzenes or any trichlorobenzenes . This restricted series of reactions is the most predominant and frequently reported pathway for the dechlorination of hexa-CB and penta-CB by enrichment cultures inoculated with either freshwater or estuarine sediments . The culture did not dechlorinate hydroxylated and methoxylated polychlorinated benzenes or a hydroxylated PCB . Bacterium DF-1 was detected by PCR/DGGE analysis following dechlorination of penta-CB but was not detected when a chlorinated benzene (CB) was not dechlorinated; detection of other members in the communitywas unaffected by the presence or absence of CB dechlorination . This is the first report of a bacterium that reductively dechlorinates both PCBs and CBs and the first identification of an organism that can dechlorinate a CB with more than four chlorines. Water Res, 2002 Jul, 36(13), 3193 - 202 Biological leaching of Mn, Al, Zn, Cu and Ti in an anaerobic sewage sludge effectuated by Thiobacillus ferrooxidans and its effect on metal partitioning; Lombardi AT et al.; The chemical fractionation and bioleaching of Mn, Al, Zn, Cu and Ti in municipal sewage sludge were investigated using Thiobacillus ferrooxidans as leaching microorganism . As a result of the bacterial activity, ORP increase and pH reduction were observed . Metal solubilization was accomplished only in experimental systems supplemented with energy source (Fe(II)) . The solubilization efficiency approached approximately 80% for Mn and Zn, 24% for Cu, 10% for Al and 0.2% for Ti . The chemical fractionation of Mn, Al, Zn, Cu and Ti was investigated using a five-step sequential extraction procedure employing KNO3, KF, Na4P2O7, EDTA and HNO3 . The results show that the bioleaching process affected the partitioning of Mn and Zn, increasing its percentage of elution in the KNO3 fraction while reducing it in the KF, Na4P2O7 and EDTA fractions . No significant effect was detected on the partitioning of Cu and Al . However, quantitatively the metals Mn, Zn, Cu and Al were extracted with higher efficiency after the bacterial activity . Titanium was unaffected by the bioleaching process in both qualitative and quantitative aspects. J Chromatogr A, 2002 Jul 19, 963(1-2), 411 - 8 Detection of explosives and their degradation products in soil environments; Halasz A et al.; Polynitro organic explosives {hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) and 2,4,6-trinitrotoluene (TNT)} are typical labile environmental pollutants that can biotransform with soil indigenous microorganisms, photodegrade by sunlight and migrate through subsurface soil to cause groundwater contamination . To be able to determine the type and concentration of explosives and their (bio)transformation products in different soil environments, a comprehensive analytical methodology of sample preparation, separation and detection is thus required . The present paper describes the use of supercritical carbon dioxide (SC-CO2), acetonitrile (MeCN) (US Environmental Protection Agency Method 8330) and solid-phase microextraction (SPME) for the extraction of explosives and their degradation products from various water, soil and plant tissue samples for subsequent analysis by either HPLC-UV, capillary electrophoresis (CE-UV) or GC-MS . Contaminated surface and subsurface soil and groundwater were collected from either a TNT manufacturing facility or an anti-tank firing range . Plant tissue samples were taken fromplants grown in anti-tank firing range soil in a greenhouse experiment . All tested soil and groundwater samples from the former TNT manufacturing plant were found to contain TNT and some of its amino reduced and partially denitrated products . Their concentrations as determined by SPME-GC-MS and LC-UV depended on the location of sampling at the site . In the case of plant tissues, SC-CO2 extraction followed by CE-UV analysis showed only the presence of HMX . The concentrations of HMX (<200 mg/kg) as determined by supercritical fluid extraction (SC-CO2)-CE-UV were comparable to those obtained by MeCN extraction, although the latter technique was found to be more efficient at higher concentrations (>300 mg/kg) . Modifiers such as MeCN and water enhanced the SC-CO2 extractability of HMX from plant tissues. Microb Ecol, 2002 Jul, 44(1), 59 - 68 Epub 2002 Apr 04. Effectiveness of municipal waste compost and its humic fraction in suppressing Pythium ultimum; Pascual JA et al.; The effect of addition of a municipal solid waste (MSW) compost and its water-soluble and humic fraction to suppress the effect of Pythium ultimum on pea plants was studied and compared with that of a chemical pesticide (metalaxyl) . The biotic and abiotic characteristics of compost involved in the biocontrol effects of these materials were also evaluated . The addition into soil of whole composts and their humic fractions reduced the effect of the pathogen on pea plants, significantly reducing the number of root lesions and Pythium populations and avoiding reductions of plant growth . The greatest pathogen suppression was achieved with the chemical pesticide . However, it also caused a significant decrease in the number of nontarget bacteria and fungi and on beneficial soil microorganisms such as Trichoderma and Pseudomonas . Addition of organic amendments increased population size of nontarget and specific biocontrol microorganisms . The humic fraction showed similar results to compost . All this suggests that metalaxyl has a nonspecific effect, producing adverse effects on aspects of soil quality . This was avoided if the chemical pesticide was reduced and replaced by organic amendments such as an MSW compost or its humic fraction. Biol Pharm Bull, 2002 Aug, 25(8), 945 - 53 Regulating factors for microglial activation; Nakamura Y; Microglia, residential macrophages in the central nervous system, can release a variety of factors including cytokines, chemokines, etc . to regulate the communication among neuronal and other types of glial cells . Microglia play immunological roles in mechanisms underlying the phagocytosis of invading microorganisms and removal of dead or damaged cells . When microglia are hyperactivated due to a certain pathological imbalance, they may cause neuronal degeneration . Pathological activation of microglia has been reported in a wide range of conditions such as cerebral ischemia, Alzheimer's disease, prion diseases, multiple sclerosis, AIDS dementia, and others . Nearly 5000 papers on microglia can be retrieved on the Web site PubMed at present (November 2001) and half of them were published within the past 5 years . Although it is not possible to read each paper in detail, as many factors as possible affecting microglial functions in in vitro culture systems are presented in this review . The factors are separated into "activators" and "inhibitors," although it is difficult to classify many of them . An overview on these factors may help in the development of a new strategy for the treatment of various neurodegenerative diseases. Orig Life Evol Biosph, 2002 Apr, 32(2), 103 - 27 D-amino acids in living higher organisms; Fujii N; The homochirality of biological amino acids (L-amino acids) and of the RNA/DNA backbone (D-ribose) might have become established before the origin of life . It has been considered that D-amino acids and L-sugars were eliminated on the primitive Earth . Therefore, the presence and function of D-amino acids in living organisms have not been studied except for D-amino acids in the cell walls of microorganisms . However, D-amino acids were recently found in various living higher organisms in the form of free amino acids, peptides, and proteins . Free D-aspartate and D-serine are present and may have important physiological functions in mammals . D-amino acids in peptides are well known as opioid peptides and neuropeptides . In protein, D-aspartate residues increase during aging . This review deals with recent advances in the study of D-amino acids in higher organisms. Dent Traumatol, 2002 Apr, 18(2), 47 - 56 The role of endodontics in the treatment of luxated permanent teeth; Barnett F; Pulp necrosis is a common complication following traumatic dental injuries and is related to the type and severity of the injury, as well as to the stage of development of the injured tooth . Endodontic intervention is required when there are clinical and radiographic signs of pulpal infection and its sequelae . Arrested tooth development with periradicular pathosis, external inflammatory root resorption, sinus tract formation and pain on percussion are indicative of root-canal infection in the post-traumatized teeth, and require immediate endodontic treatment . The use of calcium hydroxide in the treatment of teeth with post-traumatic pulp necrosis and its sequelae has been shown to be extremely beneficial for the long-term retention of the injured teeth . Calcium hydroxide has been shown to arrest and repair external inflammatory root resorptive defects, eliminate the endodontopathic microorganisms from the root canal system and induce hard-tissue barrier formation at the apex of non-vital immature teeth . This paper reviews the endodontic treatment required by post-traumatic non-vital permanent teeth. Wiad Lek, 2002, 55(3-4), 158 - 63 {Antibodies seroprevalence for mycoplasma pneumoniae antigens in patients with bronchial asthma}; Friedek D et al.; Microorganisms causing respiratory system infections, mainly viruses but also bacteria, among which there are atypical such as Chlamydophila pneumoniae, play a role in etiopathogenesis of bronchial asthma . Mycoplasma pneumoniae is suggested to take part in the initiation and the bronchial asthma exacerbation . The aim of the paper was to determine the frequency of occurrence of anti-Mycoplasma pneumoniae antibodies in serum of patients suffering from bronchial asthma in comparison with the control group of healthy persons . The presence of IgG, IgM and IgA class anti-Mycoplasma pneumoniae antibodies was assessed by immunoenzymatic assay ELISA . Serologic markers of Mycoplasma pneumoniae infection were more frequently observed in patients with bronchial asthma (15%) than in the control group (5.13%) . The diagnosis of Mycoplasma pneumoniae infection is especially important in patients with bronchial asthma . The pathogen causing bronchial hyperreactivity is eliminated by the appropriate antibiotic therapy, which allows reducing the dose of inhaled corticosteroids . The immunoenzymatic assay determining the level and class of specific antibodies to find mycoplasmatic infection quickly and precisely. Int Microbiol, 2002 Jun, 5(2), 65 - 71 Biology of killer yeasts; Marquina D et al.; Killer yeasts secrete proteinaceous killer toxins lethal to susceptible yeast strains . These toxins have no activity against microorganisms other than yeasts, and the killer strains are insensitive to their own toxins . Killer toxins differ between species or strains, showing diverse characteristics in terms of structural genes, molecular size, mature structure and immunity . The mechanisms of recognizing and killing sensitive cells differ for each toxin . Killer yeasts and their toxins have many potential applications in environmental, medical and industrial biotechnology . They are also suitable to study the mechanisms of protein processing and secretion, and toxin interaction with sensitive cells . This review focuses on the biological diversity of the killer toxins described up to now and their potential biotechnological applications. J AOAC Int, 2002 Jul-Aug, 85(4), 979 - 83 Rapid detection of single cell bacteria as a novel approach in food microbiology; D'Haese E et al.; Solid-phase cytometry (SPC) is a novel technique that allows rapid detection of bacteria at the single cell level, without the need for a growth phase . After filtration of the sample, the retained microorganisms are fluorescently labeled on the membrane filter and automatically counted by a laser scanning device . Each fluorescent spot can be visually inspected with an epifluorescence microscope connected to the ChemScan by a computer-driven moving stage . Depending on the fluorogenic labels used, information on the identity and the physiological status of the microorganisms can be obtained within a few hours . Although SPC was originally recommended for the determination of the total viable microbial count in water and other liquid samples, it may also be a promising technique for the detection and enumeration of bacteria in food samples, provided they can be isolated from the unfilterable matrix . The short detection time inherent in this approach is a considerable advantage over conventional plate counting, especially for slow-growing microorganisms . The basic principles of SPC are discussed as well as its potential for the detection of Mycobacterium paratuberculosis, a model example of a slow-growing bacterium in milk. Rev Argent Microbiol, 2002 Apr-Jun, 34(2), 57 - 65 {Inhibition of the proteolytic activity of Helicobacter pylori by mouse immune serum}; Gonzalez Segovia R et al.; The proteolytic activity (PA) of some microorganisms is an important pathogenic factor during tissue invasion . However, its role in Helicobacter pylori infection is not clear . Due to the importance of the immunological response to inhibit pathogenic factors of microorganisms, this study aims to establish an in vitro system to detect inhibitory antibodies to the PA of H . pylori in mouse serum . We obtained mouse sera from animals immunized by oral and intraperitoneal inoculations with the raw bacterial extract (BE) of H . pylori, in which we had previously detected PA . The degradation of azocasein subtract for BE was inhibited in 49.23% and 22.6% using 5 micrograms/ml of serum proteins (SP) from oral and intraperitoneal inoculation, respectively . However, when using more than 25 micrograms/ml of SP of immune serum, PA was inhibited in a similar way than with control serum . In conclusion we present a methodology for the detection of inhibitory antibodies to PA of H . pylori in the serum of the immunized mouse. Rev Argent Microbiol, 2002 Apr-Jun, 34(2), 100 - 3 {Quantitative cultures from catheters, using the Brun Buisson technique}; Soloaga R et al.; The objective of this collaborative work carried out in the Fundacion Favaloro and the Fundacion para la Lucha contra las Enfermedades Neurologicas de la Infancia, was to determine optimal conditions for incubation (time and atmosphere) of quantitative cultures of catheters processed according to the technique of vortex agitation (Brun Buisson method) . From 689 processed catheters, 551 yielded negative cultures . From the 138 positive cultures, 125 yielded monomicrobial cultures and 13 polimicrobial cultures (total number of microorganisms was 151) . In the last situation each micoorganism was considered on an individual basis . A total of 58 episodes of catheter related bacteremias occurred, being 52 monomicrobial and 6 polimicrobial (total number of microorganisms was 64) . When colony counts were compared in aerobic and in 5-10% CO2 atmospheres, a very good correlation was obtained (p = 0.27; r2 = 0.9268) . No advantage was observed by incubating plates for more than 48 hours . Colony counts performed at the second versus the third day, and at the second day versus the seventh, gave very good correlation (p = 0.10 and r2 = 0.9996; p = 0.31 and r2 = 0.9995, respectively). Curr Opin Biotechnol, 2002 Jun, 13(3), 262 - 6 Fish spoilage bacteria--problems and solutions; Gram L et al.; Microorganisms are the major cause of spoilage of most seafood products . However, only a few members of the microbial community, the specific spoilage organisms (SSOs), give rise to the offensive off-flavours associated with seafood spoilage . Combining microbial ecology, molecular techniques, analytical chemistry, sensory analysis and mathematical modelling allows us to characterise the SSOs and to develop methods to determine, predict and extend the shelf life of products. Curr Opin Biotechnol, 2002 Jun, 13(3), 213 - 7 Molecular community analysis of microbial diversity; Dahllof I; New technologies that avoid the need for either gene amplification (e.g . microarrays) or nucleic acid extraction (e.g . in situ PCR) have recently been implemented in microbial ecology . Together with new approaches for culturing microorganisms and an increased understanding of the biases of molecular methods, these techniques form the most exciting advances in this field during the past year. Rev Gastroenterol Peru, 1997 Sep, 17(3), 203 - 213 {DUODENAL PEPTIC ULCER TREATMENT BY H . PYLORI ERADICATION: CONTROLED RANDOMIZED STUDY}; Paucar H et al.; As Helicobacter pylori infection represents a very important causal factor in the development of duodenal peptic ulcer, several therapeutic trials have been developed in order to obtain an efficient eradication of the microorganism . The aim of our study has been to evaluate, in patients with duodenal peptic ulcer, the effect of double antibiotic short therapy associated to famotidine on the eradication of Helicobacter pylori, cicatrization of ulcerous lesion and the histological changes after the treatment . Forty patients with duodenal ulcer and Helicobacter pylori infection were distributed al random in two groups (I and II) . Group I received tetracycline 2 gr per day plus furazolidone 400 mg per day for a week, and famotidine 40 mg per day for four weeks . Group II received just famotidine 40 mg per day for six weeks . Al the initial endoscopy, four biopsies were taken from the gastric antrurn for histological study and determination of Helicobacter pylori . The endoscopy was repeated between 6 and 8 weeks after the beginning of the antibiotic treatment, when four biopsies were taken again from the gastric antrum with the same purpose, and the results were compared . Twenty-eight patients completed the study, 14 in group I and 14 in group II . Helicobacter pylori eradication was obtained in 85,7% of patients in group I and 0% in group II (p=0.00002) . Ulcer cicatrization was obtained in 100% of patients of group I compared to 85,7% of patients of group II (p=0.48) . The histological degree of gastritis significantly diminished after the treatment in patients of Group I, in relation to the histological parameters of severity (p=0,002), activity (p=0,00002) and presence of lymphoid follicles (p=0.02) . These changes were not significantly observed in group II . Two patients of group I had mild sideeffects (14,3%), and none of them was observed in group II (p=0.48) . We concluded that the treatment used (tetracycline + furazolidone) was highly efficacious in the eradication of Helicobacter pylori, as well as in the cicatrization, reduction of antral gastric mucous inflammation, and a low rate of side-effects. J Virol Methods, 2002 Aug, 105(1), 123 - 32 Simultaneous detection and identification of hepatitis A virus and rotavirus by multiplex nucleic acid sequence-based amplification (NASBA) and microtiter plate hybridization system; Jean J et al.; Human rotavirus and hepatitis A virus (HAV) are two of the most common causes of virus-mediated food-borne illness . Epidemiological investigations of outbreaks associated with these viruses have been hindered by the lack of available methods for their detection in foodstuffs . In this study, a multiplex nucleic acid sequence-based amplification (NASBA) system was developed to detect specifically and simultaneously human rotavirus and HAV . Two sets of primers selected from published nucleic acid sequences were used in the NASBA mixture to amplify viral RNA from both viruses . Denaturing gel electrophoresis revealed two distinct RNA products with 268 and 474 nucleotides amplified from rotavirus and HAV, respectively . The specificity of the multiplex NASBA was confirmed by a microtiter plate hybridization and detection system and by Northern blot analysis using specific oligonucleotide probes . The presence of non-homologous nucleic acid and non-target microorganisms did not have any effect on the specificity of the multiplex NASBA . Using the optimized NASBA and microtiter plate hybridization conditions, as little as 400 PFU ml x (-1) of HAV and 40 PFU ml x (-1) of rotavirus were detected . The multiplex NASBA system offers advantages over monoplex virus detection systems in terms of turnaround time and cost-effectiveness. Am J Ind Med, 2002 Jun, 41(6), 498 - 505 Exposure to culturable microorganisms in paper mills and presence of symptoms associated with infections; Haug T et al.; BACKGROUND: Previous studies of paper machine operators have to a large extent focused on endotoxins as a possible health hazard, but not culturable micro-organisms (MOs) . METHODS: Based on exposure assessment in 11 paper mills workers exposed to culturable bio-aerosols were grouped in three exposure groups . 781 exposed and 285 unexposed workers completed a questionnaire that provided data pertaining to infections and associated symptoms . RESULTS: Concentrations of culturable bacteria in process waters varied in the range 10(4)-10(6) colony forming units (cfu)/ml, and in bio-aerosols concentrations varied typically in the range 10(4)-->10(5) cfu/m3 . Operators exposed to bio-aerosols reported higher cumulative incidence of symptoms associated with infections compared to the reference population (ORs = 1.7-5.9), and the group of highest exposed workers reported higher cumulative incidence than the group of lowest exposed (ORs = 1.2-3.6) . CONCLUSION: Exposure to bio-aerosols containing culturable MOs may induce symptoms associated with infections among operators in paper mills. Clin Immunol, 2002 Jun, 103(3 Pt 1), 296 - 302 Human gamma delta T cells induce dendritic cell maturation; Ismaili J et al.; gamma delta T cells are known to be involved in the innate immune defenses against infectious microorganisms . Herein, we considered that gamma delta T cells could also influence adaptative immunity by interacting with dendritic cells (DC) in the early phase of the immune response . To investigate this hypothesis, gamma delta T cells isolated from the peripheral blood of healthy volunteers were cocultured with autologous monocyte-derived dendritic cells, which were subsequently analyzed for their expression of key surface molecules and for their production of IL-12 . First, we found that gamma delta T cells induced the upregulation of HLA-DR, CD86, and CD83 on DC . This effect did not require cell to cell contact and could be blocked by a neutralizing anti-TNF antibody . We then observed that gamma delta T cells activated by the synthetic phosphoantigen bromohydrin pyrophosphate (BrHPP) induced the production of IL-12 (p40) and IL-12 (p70) by DC, an effect that involved IFN-gamma production . The relevance of this finding to DC function was demonstrated by the increased production of IFN-gamma by alloreactive T cells when stimulated in a mixed leucocyte reaction with DC preincubated with activated gamma delta T cells . We conclude that gamma delta T cell activation might result in DC maturation and thereby in enhanced alpha beta T cell responses. Clin Infect Dis, 2002 Sep 1, 35(Suppl 1), S72 - 7 DNA probes and primers in dental practice; Conrads G; In clinical microbiology, molecular genetic techniques are increasingly being used to detect and/or differentiate uncultivable, anaerobic, or fastidious microorganisms . During the past decade, DNA probe hybridization and in vitro amplification by polymerase chain reaction have also been introduced to detect oral pathogens . The present review describes experiences with methods and commercial test systems for the detection of pathogens in periodontitis and caries. Eur J Clin Microbiol Infect Dis, 2002 Jul, 21(7), 546 - 8 Epub 2002 Jul 12. Improved decontamination method for recovering mycobacteria from patients with cystic fibrosis; Bange FC et al.; In order to improve the recovery of mycobacteria from patients with cystic fibrosis, the present study evaluated a two-step decontamination procedure for clinical specimens . A total of 920 specimens obtained from 239 patients with cystic fibrosis were treated initially with N-acetyl-L-cysteine/sodium hydroxide . Of these specimens, 31 (3.3%) showed mycobacterial growth and 415 (45.1%) remained contaminated . Contaminated specimens were then subjected to a second round of decontamination, using a combination of N-acetyl-L-cysteine/sodium hydroxide and oxalic acid . Following this second decontamination, the number of specimens overgrown by microorganisms other than mycobacteria was reduced to 7.3%, and an additional 10 specimens positive for mycobacteria were found . The results suggest this two-step protocol could improve the recovery of mycobacteria from heavily contaminated specimens. Appl Microbiol Biotechnol, 2002 Aug, 59(4-5), 523 - 8 Epub 2002 Jun 27. Comparison of green fluorescent protein expression in two industrial Escherichia coli strains, BL21 and W3110, under co-expression of bacterial hemoglobin; Kang DG et al.; Vitreoscilla hemoglobin (VHb) has been successfully used to enhance production of foreign proteins in several microorganisms including Escherichia coli . We compared the expression of an oxygen-dependent foreign protein, green fluorescent protein (GFP) under co-expression of VHb in two typical industrial E . coli strains, BL21 (a B derivative) and W3110 (a K12 derivative), which have different metabolic properties . We employed the nar oxygen-dependent promoter for self-tuning regulation of VHb expression due to the natural transition of dissolved oxygen (DO) level during culture . We observed several interesting and differing behaviors in cultures of the two strains . VHb co-expression showed a positive influence on expression, and even on solubility, of GFP in both strains; while strain BL21 had the higher GFP expression level, W3110 showed higher solubility of expressed GFP . GFP expression in strain BL21 was very largely affected by variation of aeration environments, but W3110 was not significantly impacted . We surmised that this arose from different oxygen utilization abilities and indeed the two strains showed different patterns of oxygen uptake rate . Interestingly, the VHb co-expressing W3110 strain exhibited a peculiar increasing pattern of GFP expression during the late culture period even under low aeration conditions and this enhancement was more obvious in large-scale cultures . Therefore, this strain could be successfully employed in practical large-scale production cultures where DO levels tend to be limited. Rev Soc Bras Med Trop, 2002 Jul-Aug, 35(4), 395 - 9 Clinicopathological and immunohistochemical features of the severe pulmonary form of leptospirosis; Silva JJ et al.; Four cases of severe pulmonary form of leptospirosis (SPFL) are described . In all four of these blood culture proven cases, there was severe pulmonary injury characterized by alveolar hemorrhage and acute respiratory failure . Three patients died in less than 48 hours after onset of the first respiratory signs . Leptospiral antigen detection in lung tissues was positive by immunoperoxidase in all three of these cases, suggesting that the microorganism exerts a local direct destructive action . Patients with SPFL should be carefully monitored, as the abrupt onset of severe alveolar hemorrhage can lead to respiratory insufficiency and death . The authors emphasize the importance of radiological findings and blood gas analysis for prompt clinical diagnosis, and suggest that corticosteroids, associated with antibiotics, early respiratory support, and platelet transfusions are useful as an attempt to prevent further development of SPFL. J Allergy Clin Immunol, 2002 Aug, 110(2 Suppl), S21 - 6 Blood-borne pathogens and nosocomial infections; Goldmann DA; Guidelines to prevent the transmission of blood-borne infections have evolved rapidly since the recognition that "serum hepatitis" could be transmitted to health care personnel via percutaneous exposure to blood . The HIV epidemic focused renewed attention on the problem of protecting health care personnel, culminating in "standard precautions" for patient care, which emphasized the use of gloves for all contact with blood and body fluids . This focus on protection of the health care worker sometimes obscures the other important functions of gloves: protection of patients from microorganisms on the hands of providers and prevention of patient-to-patient transmission of nosocomial pathogens . The risk of infection after percutaneous exposure to the 3 major blood-borne viruses-hepatitis B virus (HBV), hepatitis C virus (HCV), and HIV-varies greatly . The risk for a nonimmune individual exposed to HBV may be >30% if the source is Hb(e)Ag-positive . The average infection rate for HCV is 1.8% . For HIV, the average risk is 0.3%, but is higher with deep injury, when there is visible blood on the device, when a needle has been in an artery or vein, or when the source patient is in the terminal phase of HIV . Prompt administration of anti-HIV therapy reduces risk by about 80% . Mucous membrane and cutaneous exposures carry less risk . Recent efforts to reduce needlestick injuries in hospitals have reduced the risk to health care providers . Surgeons and other health care professionals who are infected with HIV or HCV pose a very small risk to their patients, although a number of outbreaks have been traced to surgeons who are HBV carriers; most have been Hb(e)Ag-positive. J Contemp Dent Pract . 2000 Nov 15;1(4):103. Strategic planning for infection control; Palenik CJ; Proper infection control procedures help to protect dental practitioners, patients, and the surrounding community . The two basic goals of dental asepsis are simple and straightforward: minimize the spread of potentially pathogenic microorganisms and neutralize organisms that have contaminated dental instruments, equipment, and other office environmental surfaces . Meeting these goals, however, can be complicated . A valuable tool would be the generation and maintenance of a written set of infection control procedures ("an office infection control manual") . One way to organize a procedures manual is to arrange required tasks in response to specific pathways of cross-infection . Also, the effectiveness of an infection control program can be positively affected by office design. J Contemp Dent Pract, 2000 Feb 15, 1(2), 28 - 41 Denture hygiene: a review and update; Shay K; Growth in the aging population has resulted in an increasing number of older persons requiring dentures . The microporous surfaces of an acrylic denture provide a wide range of environments to support microorganisms that can threaten the health of a physically vulnerable patient . The maintenance of denture prostheses is important for the health of patients and to maintain an esthetic, odor-free appliance . Mechanical, chemical, and a combination of mechanical and chemical strategies are available to patients to facilitate denture hygiene . Brushing is an ineffective method of denture disinfection . Household bleach or vinegar are effective as are the commercial, effervescent products sold for denture soaking . A new denture cleaner contains silicone polymer that provides a protective coating for dentures as a final step in the cleaning process . The coating helps to minimize the adhesion of accretions to the denture throughout the day until the next cleaning . Dental professionals must have a current knowledge of denture cleansing strategies in order to maximize the service offered to denture patients. Surg Endosc, 2002 Jul, 16(7), 1091 - 7 Epub 2002 Apr 09. Specific hygiene issues relating to reprocessing and reuse of single-use devices for laparoscopic surgery; Roth K et al.; OBJECTIVE: To determine whether reprocessed single-use devices (SUD) would (1) meet regulatory standards for sterility, and (2) meet the same material standards as new devices or if they pose an infection risk to other patients . DESIGN: The study included in the first stage single-use laparoscopic dissection devices and in the second stage a variety of clinically used and reprocessed SUDs . The suitability of these devices for cleaning, disinfection, and sterilization was examined . METHODS: Testing of cleanability was conducted on devices contaminated with radioactively labeled blood . Instruments were cleaned using hospital recommended practices . Gamma counts/second were determined before and after cleaning to localize contaminants, which were additionally visualized using light and scanning electron microscopy (SEM) . X-ray photoelectron spectroscopy (XPS) was used to quantify contamination elements on the materials tested . Residual bioburden testing on instruments contaminated with microorganisms suspended in blood prior to reprocessing was carried out to establish the efficacy of disinfection and sterilization . RESULTS: During the first stage of the study all devices remained contaminated after cleaning, but were effectively disinfected . Sterilization could not eliminate the challenge microorganisms completely . The findings during the second stage--examination of clinically used devices--were as follows: 11% of the sterile packages were damaged; 33% of the devices were incomplete and parts were missing; 54% did not meet the criteria for functionality; light microscopy, SEM, and XPS showed contamination on the outside and inside of all devices . Of the tested SUDs, 40% remained unsterile following resterilization.CONCLUSIONS: None of the reprocessed SUDs were effectively cleaned or sterilized . This may provide an opportunity for survival and growth of non-resistant or nosocomial organisms and viruses . The use of such inadequately reprocessed SUDs increases the risk for the patient, and can lead to nosocomial infection and to legal consequences for the health care facility. Clin Exp Immunol, 2002 Aug, 129(2), 359 - 69 Combination of molecular mimicry and aberrant autoantigen expression is important for development of anti-Fas ligand autoantibodies in patients with systemic lupus erythematosus; Mihara S et al.; We have reported previously that circulating anti-Fas ligand (FasL) autoantibodies able to inhibit Fas/FasL-mediated apoptosis were present in patients with systemic lupus erythematosus (SLE) . In the present study, we describe the epitopes recognized by these anti-FasL autoantibodies . Rabbit antihuman antibody, raised against a FasL fragment consisting of amino acids (aa) 103-179 (fragment 2.0), inhibited Fas/FasL-mediated apoptosis, whereas an antibody against a FasL aa 103-146 fragment (fragment 1.0) did not . This suggested that an epitope around aa 146-179 was important for Fas/FasL interaction . Epitope mapping of anti-FasL autoantibodies using deletion mutants indicated that the epitope was located around aa 163-179 . Three-dimensional molecular modelling of the Fas/FasL complex revealed that the aa 162-169 region was located on the outermost side of FasL, which suggested that the anti-FasL autoantibody would easily have access to the epitope . FasL point mutants involving aa positions 162-169 resulted in complete loss of apoptosis-inducing capability, which suggested that the aa 162-169 region was important for Fas/FasL interaction . A synthetic FasL peptide consisting of aa 161-170 blocked the binding of anti-FasL autoantibodies to FasL fragment 2.0 (aa 103-179) . The FasL aa 161-170 sequence was found to be highly homologous with aa sequences from several infectious agents . Synthetic peptides derived from some of these microorganisms cross-reacted with the epitope recognized by the autoantibodies, suggesting that several foreign infectious agent-derived proteins may share an epitope with human FasL . As lymphocytes from SLE patients aberrartly expressed FasL, it is possible that infection by one of several infectious agents may trigger cross-reactive antibody responses, after which aberrantly expressed endogenous FasL might induce the shift from a cross-reactive response to an authentic autoimmune response . Therefore, a combination of molecular mimicry and aberrant autoantigen expression may be important for the development of anti-FasL autoantibodies in SLE patients. Khirurgiia (Mosk), 2002, (7), 11 - 3 {Efficacy of clinical-serologic diagnosis of wound infection in burned patients}; Pal'tsyn AA et al.; Titer of antibodies to microorganisms obtained from wound and blood of 46 patients with burns of IIIAB--IV degree was studied . The majority of the burned patients have ability to synthesize antibodies . Agglutination of the bacteria by antibodies promoted location of infectious agent in primary focus and impeded bacteria to spread into blood and generalization of process . Not all microorganisms of wound provoke antibodies synthesis, i.e . wound infection . Therefore bacteriologic study must be supplemented by serum diagnosis . Detection of bacteria in the blood without serologic data do not permit to consider these bacteria as etiologic factor of wound infection. Biochim Biophys Acta, 2002 Jul 1, 1554(3), 202 - 11 Production and preliminary characterization of a recombinant triheme cytochrome c(7) from Geobacter sulfurreducens in Escherichia coli; Londer YY et al.; Multiheme cytochromes c have been found in a number of sulfate- and metal ion-reducing bacteria . Geobacter sulfurreducens is one of a family of microorganisms that oxidize organic compounds, with Fe(III) oxide as the terminal electron acceptor . A triheme 9.6 kDa cytochrome c(7) from G . sulfurreducens is a part of the metal ion reduction pathway . We cloned the gene for cytochrome c(7) and expressed it in Escherichia coli together with the cytochrome c maturation gene cluster, ccmABCDEFGH, on a separate plasmid . We designed two constructs, with and without an N-terminal His-tag . The untagged version provided a good yield (up to 6 mg/l of aerobic culture) of the fully matured protein, with all three hemes attached, while the N-terminal His-tag appeared to be detrimental for proper heme incorporation . The recombinant protein (untagged) is properly folded, it has the same molecular weight and displays the same absorption spectra, both in reduced and in oxidized forms, as the protein isolated from G . sulfurreducens and it is capable of reducing metal ions in vitro . The shape parameters for the recombinant cytochrome c(7) determined by small angle X-ray scattering are in good agreement with the ones calculated from a homologous cytochrome c(7) of known structure. Bioelectrochemistry, 2002 Sep, 57(2), 173 - 8 Effect of initial carbon sources on the electrochemical detection of glucose by Gluconobacter oxydans; Lee SA et al.; An electrochemical system consisted of Gluconobacter oxydans as a microorganism and 2-hydroxy-1,4-naphthoquinone (HNQ) as a mediator has been setup to examine the effect of initial carbon sources on the detection of glucose . Catalytic current due to the oxidation of glucose was observed only when both G . oxydans and HNQ were present . From amperometric measurements, it was found that the sensitivity strongly depended on the initial carbon sources . The sensitivity was highest for the cells cultured in a fructose-containing medium and decreased in the order, mannitol > sucrose > glucose > galactose > glycerol . The difference in sensitivity was explained by considering the current rising pattern at an initial stage of a microbial fuel cell constructed with the same components . The rising time, not the fuel cell efficiency, could directly be related to the sensitivity order . A sensor where G . oxydans was confined at the vicinity of the electrode by the semipermeable membrane was constructed . A linear response over a millimolar range of glucose concentration was observed with a cell grown in galactose-containing medium . This work demonstrates that the initial carbon source play an important role on glucose sensoring and should be considered in a real application. Demos, 1990, (3), 25 - 6 {It is imperative to stop environmental degradation}; Leff E; PIP: Environmental demography deals with socioenvironmental determinants of demographic processes . The fertility decline of recent years in Mexico is not associated with economic transformation, as the economic crisis, poverty, unemployment, and social inequality have been barely mitigated . Mexico is mainly urban: 51% of the population lived in cities with more than 100,000 inhabitants in 1990 taking an increasing toll on the environment . The metropolitan area of Mexico City has 50% of industrial production with 36,000 industrial plants, 2100 of all 5400 chemical plants, 22% of total population, and 25% of economically active population . There are 3 million cars . 11,000 tons of solid waste is generated daily (75% domestic and 25% industrial), and 5 million tons of contaminants are emitted annually . Sulphur, dioxide, nitrogen, ozone, and suspended particulate levels often exceed international standards . In the federal district more than 1000 squatter colonies are without drainage that affects 5 million people; and 30% of the population is without a sewage network . There are 35 cities with over 100,000 population that cannot meet their needs of drinking water with the result of a high level of gastrointestinal ailments, enteritis, diarrhea, rabies, and dengue fever caused by pathogenic microorganisms, the major cause of mortality . The degradation of the environment produces a pathological environmental emergency from chemical contamination of the air and from exposure to industrial emissions, to agrochemical products and toxic substances, to lead, insecticides, arsenic, and asbestos . Mexico has one of the highest levels of lead in the blood and DDT in mother's milk . In Mexico City lead concentration of 5 mg/cubic meter in the air, 20 mg/100 ml in maternal blood, and 13 mg/100 ml in the umbilical cord was detected associated with retardation of mental development in the 1st year of life and chronic lead poisoning . Przegl Lek, 2002, 59(2), 125 - 8 {Perioperative infections in implantative surgery . Patogenesis and prevention}; Szczepanik AM et al.; The last decades have been witnessing rapid development of the implantation surgery . The use of artificial materials to replace damaged tissues has become more and more popular . One of the complications of these procedures is graft infection . The presence of foreign body can impair local host defence on the tissue level and reduce the number of contaminating microorganisms necessary for infection to 104-105 . The most common pathogens responsible for graft infections are S . epidermidis, S . aureus and other Gram + and Gram - bacteria . The sources of infection are numerous and include patients, operative, and personnel factors . Graft-related infections are hazardous to the patients and can have even fatal consequences . Due to the limited effectiveness of applied methods to treat graft infections, more attention should be paid to prophylactic measures . These should cover all range of problems related to hospital work organisation, adequate sanitary and epidemiological conditions in the hospital wards and operating theatres as well as the use of local and systemic perioperative antibiotic prophylaxis. Biochem Biophys Res Commun, 2002 Jul 12, 295(2), 458 - 62 ORF2 gene involves in the construction of high-order structure of bacterial cellulose; Nakai T et al.; An ORF2 gene located upstream of the cellulose synthase (bcs) operon of Acetobacter xylinum BPR2001 was disrupted and a mutant (M2-2) was constructed . In static cultivation, the parent strain produced a tough, colorless, and insoluble cellulose pellicle, whereas M2-2 culture produced a thin, yellow, and fragile pellicle . The results of X-ray diffraction and 13C solid-state NMR indicated that the product of M2-2 is a mixture of cellulose I, cellulose II, and amorphous cellulose . The cellulose I to cellulose II ratio of the mixture was evaluated from the signal areas of C6 to be about 1:2 . Electron microscopy revealed that the product of M2-2 included ribbon-like cellulose and irregularly shaped particles attached to the ribbons . On the other hand, the mutant complemented with plasmid pSA-ORF2/k containing the ORF2 gene and BPR2001 produced only cellulose I . These results indicate that the ORF2 gene is involved in the production and crystallization of cellulose I microfibrils by this microorganism. Cell, 2002 Jul 26, 110(2), 191 - 202 IRAK-M is a negative regulator of Toll-like receptor signaling; Kobayashi K et al.; Toll-like receptors (TLRs) detect microorganisms and protect multicellular organisms from infection . TLRs transduce their signals through MyD88 and the serine/threonine kinase IRAK . The IRAK family consists of two active kinases, IRAK and IRAK-4, and two inactive kinases, IRAK-2 and IRAK-M . IRAK-M expression is restricted to monocytes/macrophages, whereas other IRAKs are ubiquitous . We show here that IRAK-M is induced upon TLR stimulation and negatively regulates TLR signaling . IRAK-M prevented dissociation of IRAK and IRAK-4 from MyD88 and formation of IRAK-TRAF6 complexes . IRAK-M(-/-) cells exhibited increased cytokine production upon TLR/IL-1 stimulation and bacterial challenge, and IRAK-M(-/-) mice showed increased inflammatory responses to bacterial infection . Endotoxin tolerance, a protection mechanism against endotoxin shock, was significantly reduced in IRAK-M(-/-) cells . Thus, IRAK-M regulates TLR signaling and innate immune homeostasis. Sci Total Environ, 2002 May 27, 291(1-3), 73 - 83 Cadmium biosorption by free and immobilised microorganisms cultivated in a liquid soil extract medium: effects of Cd, pH and techniques of culture; Lebeau T et al.; Instead of soil clean-up, a process not very technically and economically suited to agricultural soil contaminated by heavy metals (with a low concentration of heavy metals but highly or potentially highly contaminated surfaces), the control of the transfer of cadmium from the soil to the crops may well be a convenient method . We tested the bacterium ZAN-044, the actinomycete R27 and a basidiomycete Fomitopsis pinicola isolated for their ability to biosorb Cd, in order to inoculate agricultural soils afterwards . We then compared the cadmium biosorption by viable microbial cells which were free or immobilised in alginate beads and incubated in a soil extract liquid medium at various pH values (5, 6 and 7) and cadmium concentrations (1 and 10 mg/l) . The Cd concentration in the medium had the most important effect on the percentage of Cd biosorbed by the microorganisms, but the culture mode (free or immobilised cells) was not a side effect . In the case of F . pinicola and the actinomycete R27, the percentage of Cd biosorbed by free cells did not decrease when the Cd concentration in the medium increased (6-42% at the lowest Cd concentration to 11-48% at 10 mg Cd/l) . On the other hand, with a low Cd concentration (1 mg Cd/l), the percentage of Cd biosorbed by the bacterium ZAN-044 was maximum (69%) at pH 7, while this bacterium did not grow at 10 mg Cd/l and it did not accumulate Cd . For the three micro-organisms tested, relatively low specific biosorptions of Cd were observed, when the microorganisms were cultivated with a soil extract medium ('poor' medium), comparatively to those with a 'rich' medium . Finally, the choice of microorganism for the inoculation of contaminated soils depends on the cadmium level in the medium and on the distribution of the metal between the biomass and the medium. J Clin Microbiol, 2002 Aug, 40(8), 2725 - 8 PCR-based method for detecting viral penetration of medical exam gloves; Broyles JM et al.; The test approved by the U.S . Food and Drug Administration for assessment of the barrier quality of medical exam gloves includes visual inspection and a water leak test . Neither method tests directly the ability of gloves to prevent penetration by microorganisms . Methods that use microorganisms (viruses and bacteria) to test gloves have been developed but require classical culturing of the organism to detect it . We have developed a PCR assay for bacteriophage phiX174 that allows the rapid detection of penetration of gloves by this virus . The method is suitable for use with both latex and synthetic gloves . The presence of glove powder on either latex or synthetic gloves had no effect on the ability of the PCR assay to detect bacteriophage DNA . The assay is rapid, sensitive, and inexpensive; requires only small sample volumes; and can be automated. Int J Syst Evol Microbiol, 2002 Jul, 52(Pt 4), 1277 - 83 Thalassospira lucentensis gen . nov., sp . nov., a new marine member of the alpha-Proteobacteria; Lopez-Lopez A et al.; A novel bacterium from the Mediterranean Sea was isolated under oligotrophic conditions at in situ temperature after prolonged continuous culture . The isolates were initially characterized by partial 16S rRNA gene sequencing . Similarity searches of one of the isolates, QMT2T, indicated high sequence identity to the well-characterized Rhodospirillum rubrum, {Aquaspirillum} itersonii and {Oceanospirillum} pusillum micro-organisms, which are representatives of the alpha-subclass of the Proteobacteria . The highest level of similarity of the complete 165 rRNA gene with respect to these microorganisms was 89% . Features such as the low similarities of 165 rRNA of QMT2T with its phylogenetically close neighbours, the distinct G+C content, and the differences in phenotypic features, including pigmentation, fatty acid composition, salt tolerance, the lack of bacteriochlorophyll a, and the capacity to use carbohydrates as carbon sources, are indicative of the novel nature of the isolate QMT2T among the alpha-Proteobacteria . This report describes the classification of strain QMT2T (= DSM 14000T = CECT 5390T) as a new genus and species, Thalassospira lucentensis gen . nov, sp . nov., in the family Rhodospirillaceae. Annu Rev Phytopathol, 2002, 40, 309 - 48 Epub 2002 May 13. Microbial populations responsible for specific soil suppressiveness to plant pathogens; Weller DM et al.; Agricultural soils suppressive to soilborne plant pathogens occur worldwide, and for several of these soils the biological basis of suppressiveness has been described . Two classical types of suppressiveness are known . General suppression owes its activity to the total microbial biomass in soil and is not transferable between soils . Specific suppression owes its activity to the effects of individual or select groups of microorganisms and is transferable . The microbial basis of specific suppression to four diseases, Fusarium wilts, potato scab, apple replant disease, and take-all, is discussed . One of the best-described examples occurs in take-all decline soils . In Washington State, take-all decline results from the buildup of fluorescent Pseudomonas spp . that produce the antifungal metabolite 2,4-diacetylphloroglucinol . Producers of this metabolite may have a broader role in disease-suppressive soils worldwide . By coupling molecular technologies with traditional approaches used in plant pathology and microbiology, it is possible to dissect the microbial composition and complex interactions in suppressive soils. Sci Total Environ, 2002 Jun 26, 292(3), 215 - 29 Soiling and decay of N.M.E.P . limestone tablets; Viles HA et al.; The British National Materials Exposure Programme (N.M.E.P.) ran from 1987 to 1995 and involved exposure of a range of materials samples (including tablets of Monks Park and Portland Limestones) at over 20 sites around Britain for 1-, 2-, 4- and 8-year periods, under known climate and pollution conditions . Deterioration of the limestone tablets has previously been recorded in terms of weight change, contents of soluble salts, and visual soiling . In the present study samples from exposed and sheltered positions at Wells, Bolsover and Lough Navar have been studied using a spectrophotometer, optical microscopy and scanning electron microscopy (SEM) to investigate the distribution and nature of particulate material and its role in soiling and decay . Clearly, recognisable pollutant particles such as perforated cenospheres, are only rarely present . Organisms and organic remains, including filamentous microorganisms and pollen grains, are widely distributed . At each site, soiling has different characteristics in terms of composition and change over time related in part to differences in climate and pollution histories . There is no general link between degree of soiling and amount of decay (in terms of surface recession) as the nature of decay is a key influence on the relation between soiling and decay. Huan Jing Ke Xue, 2002 May, 23(3), 49 - 52 {Effect of the COD and total phosphorus concentration on biological phosphorus removal supplied with acetate as a sole carbon source}; Ruan W et al.; The effect of the concentration of acetate and total phosphorus on biological phosphorus removal process in sequencing batch reactor fed with sodium acetate (as sole carbon source) was studied in this paper . The reason of the inefficiency of phosphorus removal by sludge fed with high concentration acetate solution was analyzed . The results indicated that when COD < 600 mg.L-1 the phosphorus removal increased with the increase of COD/TP; and this removal increased obviously when COD/TP < 50 but not when COD/TP > 50 . The phosphorus removal descended at high acetate concentration (COD > 600 mg.L-1) and broke down when COD > 1000 mg.L-1 . It is the transmission of high acetate concentration from the anaerobic stage to the aerobic stage that led to the proliferation of the filamentous microorganism and the expansion of sludge, then polyP bacteria was wash out, which resulted in the drop of phosphorus removal. Eur Surg Res, 2002 Jul-Aug, 34(4), 275 - 84 Phagocytosis by emigrated, intra-abdominal neutrophils is depressed during human secondary peritonitis; Holzer K et al.; The phagocytic function of neutrophils is a crucial element in host defense against invading microorganisms . Patients with diffuse peritonitis depend on adequate reactivity of neutrophils, in particular locally in the peritoneal cavity as well as in the circulation . This study examined phagocytosis as well as numerical expression of Fcgamma I-III (CD16, CD32, CD64) and complement receptors (CD18, CD35) of emigrated, intra-abdominal and circulating neutrophils during human secondary peritonitis using fluorescence-activated cell analysis . Optimally opsonized E . coli bacteria were used independently of the well-known low level of opsonic molecules during peritonitis . Compared with controls (abdominal surgery without peritonitis), the percentage of emigrated neutrophils which engulfed E . coli bacteria was significantly depressed until 48 h after diagnosis of, and surgery for, peritonitis . When patients with complicated peritonitis (septic shock, multiple organ failure) were compared with patients without complications, phagocytosis was even more depressed in patients with complications . Numerical expression of CD64 (Fcgamma RI) and CD35 (CR1) increased significantly on emigrated polymorphonuclear leukocytes (PMNs) during peritonitis when compared to controls . There was no difference in CD18 and CD32 (Fcgamma RII) expression between the two groups . Numerical expression of CD16 (Fcgamma RIII) on emigrated PMNs decreased significantly in peritonitis . This was more pronounced in patients with complicated peritonitis . We conclude that there is a long-lasting depression of phagocytosis by emigrated PMNs during peritonitis, independent of the opsonic activity . Our data suggest that decreased phagocytosis might be correlated to the profound drop in CD16 on these cells . Allergy, 2002, 57 Suppl 72, 75 - 8 Role of complement in xenotransplantation; Mollnes TE et al.; The xenotransplantation research is driven by the increasing gap between the number of patients with end-stage organ failure on waiting lists for transplantation and the supply of allografts . The lack of success in developing suitable artificial organs for permanent treatment of organ failure has further strengthened the need for xenotransplantation research . Pigs are now generally accepted to be the source animal of choice . Transplantation of pig organs to humans faces several barriers which have to be overcome before it comes to clinical application: (1) anatomical and physiological conditions; (2) immunological rejection mechanisms; (3) molecular compatibility between signal molecules of the two species; (4) risk of transmission of microorganisms, particularly pig endogenous retroviruses; and (5) legal and ethical aspects both with respect to the animal and the recipient . Here we will focus on the role of the complement system in the rejection of immediately vascularized pig-to-primate xenografts . The hyperacute rejection occurring within minutes after transplantation is mediated by binding of natural antibodies to the Galalpha(l-3)Gal epitope on the endothelial cells with subsequent complement activation . Whereas inhibition of complement activation protects against hyperacute rejection, the role of complement in the later rejection phases is less clarified. CLAO J, 2002 Jul, 28(3), 146 - 8 Infectious keratitis: correlation between corneal and contact lens cultures; Martins EN et al.; PURPOSE: This article reports a study of the effectiveness of performing contact lens cultures to identify causative organisms of infectious keratitis in patients using extended-wear soft contact lenses (cosmetic or therapeutic) . METHODS: Patients' records from the Ocular Microbiology Laboratory of the Department of Ophthalmology, Paulista School of Medicine, from 1976 to 2000, and data of patients with contact lens-related keratitis in which the contact lens had been cultured were collected and reviewed . RESULTS: One hundred thirteen patients were included . Only soft contact lenses had been cultured and 29.2% of them were therapeutic contact lenses . An overall concordance of 84.0% was found between cultures obtained from corneal scrapings and contact lenses, with a higher rate in fungal keratitis (100%) compared to amebic (80.0%) and bacterial (74.5%) keratitis . In cases of concomitant bacterial growth in cultures of scrapings and contact lenses, the same organism was detected in 97.04% . Most cases of bacterial infection were caused by Pseudomonas spp . Among the discordant cases (16%), a microorganism was more likely to be isolated by contact lens culture (94.4%) . CONCLUSION: Contact lens cultures may identify the causative organisms in most cases of contact lens-related keratitis . The clinical relevance of positive contact lens cultures in keratitis with negative corneal cultures needs further investigation. J Biotechnol, 2002 Sep 25, 98(2-3), 227 - 41 DNA chips for yeast biotechnology . The case of wine yeasts; Perez-Ortin JE et al.; The yeast Saccharomyces cerevisiae is one of the most popular model organisms . It was the first eukaryote whose genome was sequenced . Since then many functional analysis projects have tried to find the function of many genes and to understand its metabolism in a holistic way . Apart from basic science this microorganism is of great interest in several biotechnology processes, such as winemaking . Only global studies of the cell as a whole can help us to understand many of the technical problems facing winemaking . DNA chip technology is one of the most promising tools for the analysis of cell physiology . Yeast has been the model organism for the development of this technique . Many of the studies can be applied to improve our knowledge of wine strains . Nevertheless wine strains are quite different in some aspects from the laboratory reference strains so a particular study of wine strains and especially during the winemaking process is needed . During the past two years some groups have started this study and the first results have been published . We review here the current state of the knowledge of wine yeast and the capacity of DNA chip technology for its improvement. Dig Dis Sci, 2002 Jul, 47(7), 1638 - 43 Helicobacter infection in patients with HCV-related chronic hepatitis, cirrhosis, and hepatocellular carcinoma; Dore MP et al.; Chronic hepatitis may progress to cirrhosis and hepatocellular carcinoma (HCC) . HCC represents one of the most common human cancers . Incidence rates for this tumor vary widely on a worldwide, suggesting that environmental factors such as infectious microorganisms, carcinogens, or nutrition play a role in its pathogenesis . Several Helicobacter spp . colonize the liver of animals and induce hepatitis . The aim of this study was to determine whether Helicobacter infection was associated with HCV-related liver diseases in humans . Liver tissue samples, including biopsy and surgically excised tissues, were collected from patients positive for hepatitis C viruses (HCV) RNA in the serum . Genomic DNA was extracted from sections of formalin-fixed paraffin-embedded tissues by using the QIAamp Tissue Kit and subjected to polymerase chain reaction (PCR) analysis using two sets of Helicobacter-specific 16S ribosomal RNA primers . To identify positive samples for H . pylori, a set of primers specific for a conserved region in the H . pylori vacA gene were also used . The patients' H . pylori status was determined by ELISA . Forty-one patients (mean age 54.9, range 19-78 years; 24 men) were studied . Thirty patients had chronic viral hepatitis (CH) without (N = 18) or with (N = 12) cirrhosis (CIR), and 11 patients had HCC . Anti-H . pylori IgG was detected in 54% . The expected 422- and 210-bp fragments of Helicobacter 16S rRNA were amplified from 27% of liver samples, including 17% of CH-CIR and 55% of HCC (P = 0.004) . The vacA sequence was amplified in 10 of 41(24%) samples (27% of those with HCC) . These data confirm the presence of H . pylori DNA sequences in human liver and suggest an association of Helicobacter spp . with HCV-related chronic liver diseases . Further studies are needed to ascertain whether Helicobacter spp . infection plays a role in the development of HCC. Dig Dis Sci, 2002 Jul, 47(7), 1447 - 57 Increased proliferation and apoptosis of colonic epithelial cells in dextran sulfate sodium-induced colitis in rats; Vetuschi A et al.; We have evaluated morphologic alterations and epithelial cell apoptosis and proliferation of colonic mucosa in the acute and chronic phases of DSS-induced colitis . Colitis was induced in Sprague-Dawley rats by 7 days of 4% DSS oral administration followed by 7 days of tap water for one, two, and three cycles . Control rats receved tap water only . Morphological changes in colonic mucosa were evaluated and scored by light and scanning electron microscopy . Apoptosis was studied by TUNEL assay and cell proliferation by Ki-67 immunoreaction . The expression of both proapoptotic (Fas, FasL, Bax, p53) and antiapoptotic (Bc12) cellular proteins was determined by immunohistochemistry . Morphologic assessment showed the most severe colonic epithelial lesions and inflammation in the distal colon with a trend to increasing severity from the first to the third DSS cycle . In DSS rats, the epithelial apoptotic index increased 20-fold after the first cycle and 120-fold after the second and third cycles compared with the controls; in the same way, the expression index of proapoptotic proteins (Fas, FasL, Bax, p53) dramatically increased . The proliferative index increased about 40 to 60-fold compared to controls, with no difference among the three DSS cycles . In conclusion, DSS-induced colitis in rats, which has many structural and ultrastructural features similar to those seen in human ulcerative colitis, is a suitable model for studying increased epithelial apoptosis and proliferation . Further studies employing this model will permitt two hypotheses to be tested . (1) Increased apoptosis may lead to a breakdown of the epithelial barrier function and facilitate the mucosal invasion of intraluminal microorganisms and/or antigens . (2) Abnormal and persistent epithelial hyperproliferation could be causally related to the development of colorectal cancers in the setting of chronic colonic inflammation. Environ Sci Technol, 2002 Jul 15, 36(14), 3170 - 5 Fulvic acid oxidation state detection using fluorescence spectroscopy; Klapper L et al.; Humic substances are a heterogeneous class of moderate molecular weight, yellow-colored biomolecules present in all soils, sediments, and natural waters . Although humic substances are generally resistant to microbial degradation under anaerobic conditions, some microorganisms in soils and sediments can use quinone moieties in humic substances as electron acceptors . Laboratory experiments have shown that humic substances can act as electron shuttles in the microbial reduction of ferric iron . Field studies of electron shuttling processes have been constrained by the lack of methods to characterize the oxidation state of quinone moieties in humic substances at natural concentrations . All humic substances have fluorescent properties, and fluorescence spectroscopy can indicate differences in precursor organic source of humic substances . Here we show that the quinone moieties responsible for electron transfer reactions contribute significantly to the fluorescence of humic substances . Further we use fluorescence spectroscopy to elucidate the oxidation state of quinone moieties in humic substances at natural concentrations found in sediment interstitial waters. Br J Haematol, 2002 Aug, 118(2), 654 - 62 Induction and characterization of cutaneous lymphocyte antigen on natural killer cells; Tsuchiyama J et al.; Cutaneous lymphocyte antigen (CLA) has been reported to be expressed mainly by memory/effector T lymphocytes infiltrating inflammatory skin lesions and cutaneous T-cell lymphoma . It has been suggested that CLA is a specific homing receptor, facilitating the T-cell migration into skin lesions, and also an indicator of the skin-homing T-cell subset . In the present study, we investigated the expression of CLA in natural killer (NK) cells defined phenotypically as surface CD3- and CD56+ cells in peripheral blood . CLA was definitely expressed on CD3-CD56+ cells at a level comparable to CD3+ cells in peripheral blood of normal Japanese volunteers . After in vitro stimulation of peripheral blood mononuclear cells with interleukin 2 (IL-2) and IL-12, there was a significant increase in the number and percentage of CLA+ NK cells but not CLA+ T cells (P < 0.01) . To analyse the characteristics of CLA expressed by NK cells, we investigated a CLA+ NK-leukaemia cell line, NK-YS, established from a patient with NK leukaemia/lymphoma with skin infiltration . In the in vitro study, the CLA-expressing NK-leukaemic cell line bound to E-selectin-transfected cells and was inhibited by HECA 452 antibody or neuraminidase treatment of leukaemic cells . These findings suggest that CLA expressed by NK cells is a homing receptor for the E-selectin molecule and may explain skin infiltration by NK cells and NK lymphoma cells analogous to T cells . An NK-cell subset expressing CLA must play an important role in host defence against microorganisms and neoplasms in skin lesions. Biochemistry (Mosc), 2002 Jul, 67(7), 826 - 35 Novel class of polytopic proteins with domains associated with putative protease activity; Grigorenko AP et al.; A significant proportion of early onset Alzheimer's disease (AD) is caused by mutations in human genes for amyloid precursor protein (APP), presenilins 1 and 2 (PSEN1,2) . AD associated mutations in PSEN1,2 genes alter the gamma-secretase cleavage activity of APP resulting in increased production of amyloidogenic Abeta42 . PSEN dependent intramembrane proteolysis was described as an important step required for cleavage of Notch receptors, Notch-dependent signal transduction, and processing of other proteins . It is still unclear whether presenilins are unusual intramembrane proteases or they are necessary cofactors of gamma-secretase cleavage of APP and Notch . Identification of other proteins similar to presenilins may resolve this dilemma . We describe here the identification of novel families of genes encoding polytopic transmembrane proteins of Eukaryotes (IMPASes) and Arachaea(membrases) . These proteins have a predicted structure similar to presenilins . The amino acid similarity is significant in domains carrying invariant amino acid residues, which are critical in specific presenilin-regulated endoproteolysis . Many members of the IMPAS family have protease associated domains (PA) typical of proteases . We identified and cloned five human IMPAS genes . Expression analysis of the hIMP1 gene (located on chromosome 20) was performed in human cell tissues and transfected cell cultures . The data demonstrate that a conservative class of putative protease-related polytopic proteins related to presenilins exists in multicellular eukaryotes and microorganisms. Anal Chem, 2002 Jul 15, 74(14), 3336 - 41 Biosensor for asparagine using a thermostable recombinant asparaginase from Archaeoglobus fulgidus; Li J et al.; Asparaginase from the hyperthermophilic microorganism Archaeoglobus fulgidus was cloned and expressed in Escherichia coli as a fusion protein with a polyhistidine tail . After heat treatment to denature most of the native E . coli proteins, the enzyme was purified by an immobilized metal ion affinity chromatography method . The activity of the enzyme was determined by monitoring the change in ammonium concentration in solution . It was found that the enzyme is thermostable at temperatures as high as 85 degrees C . The KM for L-asparagine was 8 x 10(-5) and 5 x 10(-6) M at 37 and 70 degrees C, respectively . The catalytic activity for L-asparagine was 5-fold higher than for D-asparagine . The enzyme was immobilized in front of an ammonium-selective electrode and used to develop a biosensor for asparagine . The biosensor had a detection limit of 6 x 10(-5) M for L-asparagine . Unlike a sensor based on asparaginase from E . coli, the biosensor based on recombinant asparaginase from A . fulgidus demonstrated higher stability. J Antibiot (Tokyo), 2002 May, 55(5), 480 - 94 Cyclipostins, novel hormone-sensitive lipase inhibitors from Streptomyces sp . DSM 13381 . II . Isolation, structure elucidation and biological properties; Vertesy L et al.; Hormone-sensitive lipase (HSL) is a key enzyme of lipid metabolism and its control is therefore a target in the treatment of diabetes mellitus . Cultures of the Streptomyces species DSM 13381 have been shown to potently inhibit HSL . Ten inhibitors of HSL, termed cyclipostins, have been isolated from the mycelium of this microorganism and a further nine related compounds detected . Their structures were characterized by 2-D NMR experiments and by mass spectrometry and were found to comprise neutral cyclic enol phosphate esters with an additional y-lactone ring . On account of their ester-bound fatty alcohol side chain, the cyclipostins have physicochemical properties similar to those of triglycerides . The outstanding characteristic of the cyclipostins is their strong anti-HSL activity, with IC50 values in the nanomolar range. Middle East J Anesthesiol, 2002 Jun, 16(5), 529 - 34 An alternative simple method in laryngoscope blade decontamination; Orhan ME et al.; The cleaning and disinfection of laryngoscope blades is controversial . The aim of this study is to investigate the efficacy of two different chemical disinfectant agents and tap water where the laryngoscope blades were contaminated by different microorganisms and try to create a simple, effective and easy decontamination method . The results of our study demonstrate that the decontamination of the laryngoscope blades, which are cleansed with tap water, is not a reliable approach . In conclusion, mechanical cleaning of blades with water and the immersion in 2% glutaraldehyde or 10% polyvinyl pyrrolidine iodine for 10 minutes is an effective method for decontamination of laryngoscope blades. Gene, 2002 Jun 26, 293(1-2), 161 - 8 Differences in expression of two endoxylanase genes (xynA and xynB) from Penicillium purpurogenum; Chavez R et al.; A number of xylanolytic microorganisms secrete to the medium several molecular forms of endoxylanases . The physiological function of these isoforms is not clear; one possibility is that they are produced under different growth conditions . To study this problem, we have used two endoxylanases (XynA and XynB) produced by the fungus Penicillium purpurogenum . These enzymes have been previously purified and characterized; they belong to family 10 and 11 of the glycosyl hydrolases, respectively . The promoters of the xynA and xynB genes have been sequenced; both present consensus sequences for the binding of the carbon catabolite repressor CreA, but otherwise show substantial differences . The xynB promoter has eight boxes in tandem for the binding of the XlnR activator and lacks the consensus sequence for the PacC pH regulator . On the other hand, the xynA promoter contains one XlnR box and three PacC consensus sequences . To investigate if these differences are reflected in gene expression, Northern blot assays were carried out . The xynA gene is transiently expressed when oat spelt xylan is used as carbon source, but negligible expression was observed with birchwood xylan, xylose or xylitol . In contrast, xynB is broadly induced by all these carbon sources; this may be related to the presence of several XlnR boxes . Similar results were obtained by zymogram analysis of the expressed proteins . The different induction capabilities of birchwood and oat spelt xylan may be due to differences in their composition and structure . Expression assays carried out at different pH reflects that, despite the lack of PacC binding sites in the xynB promoter, this gene is tightly regulated by pH . The findings described here illustrate new and important differences between endoxylanases from families 10 and 11 in P . purpurogenum . They may help explain the production of multiple endoxylanase forms by this organism. Mediators Inflamm, 2002 Jun, 11(3), 149 - 53 Effects after inhalation of (1-->3)-beta-D-glucan and relation to mould exposure in the home; Beijer L et al.; BACKGROUND: Damp conditions indoors favour the growth of microorganisms, and these contain several agents that may cause inflammation when inhaled . Moulds contain a polyglucose in their cell wall, defined as (1-->3)-beta-D-glucan, exhibiting effects on inflammatory cells . AIM: The aim of the present study was to evaluate whether an inhalation challenge to purified (1-->3)-beta-D-glucan (grifolan) in humans could induce effects on inflammatory markers in blood, and to evaluate whether the reactions were related to the home exposure to (1-->3)-beta-D-glucan . METHODS: Seventeen subjects in homes with high levels of airborne (1-->3)-beta-D-glucan (G-high) and 18 subjects in homes with low levels of (1-->3)-beta-D-glucan (G-low) underwent two randomised, double-blind inhalation challenges, one to (1-->3)-beta-D-glucan suspended in saline and one to saline alone . A blood sample was taken before and after the challenges, and differential cell count, granulocyte enzymes in serum and the secretion of cytokines from peripheral blood mononuclear cells (PBMC) were measured . RESULTS: Inhalation challenge with (1-->3)-beta-D-glucan induced a decrease in the secretion of tumour necrosis factor alpha from endotoxin-stimulated PBMC in the G-high group as well as in the G-low group . In the G-high group, the inhalation of (1-->3)-beta-D-glucan induced an increase in blood lymphocytes that was significantly different from the saline-induced effect . CONCLUSIONS: The results suggest that an inhalation challenge to (1-->3)-beta-D-glucan has an effect on inflammatory cells and this effect may be related to a chronic exposure to moulds at home. Chemosphere, 2002 Jul, 48(1), 133 - 8 Integrated assessment for aerobic biodegradability of organic substances; Jiang Z et al.; In this paper, the amount of oxygen consumption, end products and activities of microorganisms are considered as the three factors which affect the biodegradability of organic substances in water . Two integrated assessment methods for biodegradability of organic substances, fuzzy clustering integrated assessment and weighted integrated assessment, have been developed . Simultaneously, the detail steps for assessing a new organic substance biodegradability with these two integrated assessment methods are proposed. Acta Crystallogr D Biol Crystallogr, 2002 Aug, 58(Pt 8), 1374 - 5 Epub 2002 Jul 20. Crystallization and preliminary X-ray crystallographic studies of rice alpha-galactosidase; Fujimoto Z et al.; alpha-Galactosidases catalyze the hydrolysis of galactooligosaccharides and galactopolysaccharides to alpha-galactose residues and are widely distributed in microorganisms, plants and animals . alpha-Galactosidase from rice (Oryza sativa L . ssp . japonica) was crystallized by the hanging-drop vapour-diffusion method . The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 63.1, b = 71.3, c = 85.6 A, and diffract beyond 1.9 A resolution. Int J Antimicrob Agents, 2002 Jun, 19(6), 466 - 70 The role of Chlamydia trachomatis in prostatitis; Weidner W et al.; Ascending chlamydial infections have been thought to be an infective cause of prostatitis for the last three decades . Unfortunately, the definitive association between isolation of an infective agent and its prostatic origin is limited by various factors, although modern techniques of molecular biology for identification of the microorganisms are available . Two major problems are: (1) diagnostic material passing the urethra may reflect only urethral contamination, (2) prostatic biopsy specimens from the gland may also contain urethral material . The ejaculate has the same limitations, and an ideal test for detection of Chlamydia species in ejaculate specimens is not available yet . Investigations for local chlamydial IgA-antibodies may be useful; the overlap with Chlamydia pneumoniae and Chlamydia psittaci means a clear differentiation on an type-specific basis is necessary, which is normally provided by the 'elaborate' microimmunofluorescence test . Modern p-ELISAs using major outer membrane protein parts as antigens may deliver identical results in the future . In the follow-up of standardized prostatitis patients, a combination of such urological tests in EPS and seminal plasma combined with genital chlamydial DNA material, may further elucidate the chlamydial aetiology of prostate infection. Vet Rec, 2002 Jun 22, 150(25), 782 - 4 Food safety on the slaughterline: inspection of pig heads; Petersen JV et al.; This paper summarises information on the current inspection procedures for pig heads on the slaughterline and their impact on food safety, and considers the implications for food safety of certain lesions . It is argued that although a modified slaughter and inspection technique would decrease the contamination of the carcase with pathogenic microorganisms, leaving lesions in the head undiscovered would be of little or no importance either for human health or for the overall supervision of animal health. J Soc Biol, 2002, 196(1), 37 - 46 {Regulation of human neutrophil oxidative burst by pro- and anti-inflammatory cytokines}; Gougerot-Pocidalo MA et al.; Human polymorphonuclear neutrophils play a key role in host defenses against invading microorganisms . In response to a variety of stimuli, neutrophils release large quantities of superoxide anion (O2.-) in a phenomenon known as the respiratory burst . O2.- is the precursor of potent oxidants, which are essential for bacterial killing and also potentiate inflammatory reactions . Regulation of this production is therefore critical to kill pathogens without inducing tissue injury . Neutrophil production of O2.- is dependent on the respiratory burst oxidase, or NADPH oxidase, a multicomponent enzyme system that catalyzes NADPH-dependent reduction of oxygen to O2.- . NADPH oxidase is activated and regulated by various neutrophil stimuli at infectious or inflammatory sites . Proinflammatory cytokines such as GM-CSF, TNF and IL-8 modulate NADPH oxidase activity through a priming phenomenon . These cytokines induce a very weak oxidative response by PMN but strongly enhance neutrophil release of reactive oxygen species on exposure to a secondary applied stimulus such as bacterial N-formyl peptides . Priming phenomena are involved in normal innate immune defense and in some inflammatory diseases . The mechanisms underlying the priming process are poorly understood, although some studies have suggested that priming with various agonists is regulated at the receptor and post-receptor levels . Resolution of inflammation involves desensitization phenomena and cytokines are involved in this process by various mechanisms . A better understanding of phenomena involved in the regulation of NADPH oxidase could help to develop novel therapeutic agents for inflammatory diseases involving abnormal neutrophil superoxide production. J Histochem Cytochem, 2002 Aug, 50(8), 1081 - 9 Cellular expression of gut chitinase mRNA in the gastrointestinal tract of mice and chickens; Suzuki M et al.; Recently, the second mammalian chitinase, designated acidic mammalian chitinase (AMCase), has been identified in human, mouse, and cow . In contrast to the earlier identified macrophage-derived chitinase (chitotriosidase), this chitinase is richly expressed in the gastrointestinal (GI) tract, suggesting its role in digestion of chitin-containing foods as well as defense against chitin-coated microorganisms and parasites . This in situ hybridization study first revealed cellular localization of the gut-type chitinase in the mouse and chicken . In adult mice, the parotid gland, von Ebner's gland, and gastric chief cells, all of which are exocrine cells of the serous type, expressed the gut chitinase mRNA . In the chicken, oxyntico-peptic cells in glandular stomach (proventriculus) and hepatocytes expressed the chitinase mRNA . Because cattle produce the gut chitinase (chitin-binding protein b04) only in the liver, the gut chitinases in mammals and birds have three major sources of production, i.e., the salivary gland, stomach, and liver . During ontogenetic development, the expression level in the parotid gland and stomach of mice increased to the adult level before weaning, whereas in the stomach of chickens intense signals were detectable in embryos from incubation day 7. Klin Lab Diagn, 2002 Jun, (6), 38 - 9 {Use of specific antibodies labelled with colloid silver particles for detection of Brucella antigens with dot immune analysis}; Zagoskina TIu et al.; Fitness of dot immuno-analysis for detection of Brucella antigens labeled with colloid silver is evaluated . Soluble lipopolysaccharides and protein-saccharide antigen and corpuscular antigens of 22 Brucella strains (7 species) pathogenic for humans and animals in the S and R forms were used . The specificity of the method was tested on 10 heterologous microorganisms whose antigens were closely related . The suggested test system is simple, economic, highly sensitive (from 62 thousands to 8 million CFU/ml) and specific, requires no expensive equipment, and is an alternative to enzyme immuno-assay and dot immuno-analysis with gold immunosole. Rev Environ Contam Toxicol, 2002, 174, 49 - 170 Physical and chemical properties of pyrethroids; Laskowski DA; The physical and chemical properties of the pyrethroids bifenthrin, cyfluthrin, cypermethrin (also zetacypermethrin), deltamethrin, esfenvalerate (also fenvalerate), fenpropathrin, lambda-cyhalothrin (also cyhalothrin), permethrin, and tralomethrin have been reviewed and summarized in this paper . Physical properties included molecular weight, octanol-water partition coefficient, vapor pressure, water solubility, Henry's law constant, fish biocencentration factor, and soil sorption, desorption, and Freundlich coefficients . Chemical properties included rates of degradation in water as a result of hydrolysis, photodecomposition, aerobic or anaerobic degradation by microorganisms in the absence of light, and also rates of degradation in soil incubated under aerobic or anaerobic conditions . Collectively, the pyrethroids display a highly nonpolar nature of low water solubility, low volatility, high octanol-water partition coefficients, and have high affinity for soil and sediment particulate matter . Pyrethroids have low mobility in soil and are sorbed strongly to the sediments of natural water systems . Although attracted to living organisms because of their nonpolar nature, their capability to bioconcentrate is mitigated by their metabolism and subsequent elimination by the organisms . In fish, bioconcentration factors (BCF) ranged from 360 and 6000 . Pyrethroids in water solution tend to be stable at acid and neutral pH but {table: see text} become increasingly susceptible to hydrolysis at pH values beyond neutral . Exceptions at higher pH are bifenthrin (stable), esfenvalerate (stable), and permethrin (half-life, 240 d) . Pyrethroids vary in susceptibility to sunlight . Cyfluthrin and tralomethrin in water had half-lives of 0.67 and 2.5 d; lambda-cyhalothrin, esfenvalerate, deltamethrin, permethrin, and cypermethrin were intermediate with a range of 17-110 d; and bifenthrin and fenpropathrin showed the least susceptibility with half-lives of 400 and 600 d, respectively . Pyrethroids on soil can also undergo photolysis, often at rates similar to that in water . Half-lives ranged from 5 to 170 d . {table: see text} Pyrethroids are degradable in soils with half-lives ranging from 3 to 96 d aerobically, and 5 to 430 d anaerobically . For those pyrethroids studied in water (cypermethrin, deltamethrin, esfenvalerate, fenpropathrin, and lambda-cyhalothrin), aerobic and anaerobic degradation often continued at rates similar to that displayed in soil. FEMS Microbiol Lett, 2002 Jul 16, 213(1), 133 - 9 Phospholipid etherlipid and phospholipid fatty acid fingerprints in selected euryarchaeotal monocultures for taxonomic profiling; Gattinger A et al.; Phospholipid etherlipid (PLEL) derived isoprenoids and phospholipid fatty acids (PLFA) were determined in eight Euryarchaeotal monocultures for taxonomic profiling . For the first time significant amounts of fatty acids in the PLFA of Euryarchaeota were determined . The PLFA proportion varied between 11.3 and 35.5% of the total phospholipid side chains except in Methanothermus fervidus where PLFA accounted for 89.0% of the total phospholipid side chains . Fractionation of fatty acids prior to gas chromatography mass spectrometry analysis revealed that non-ester-linked fatty acids dominated which accounted for 85.5-95.2% of total PLFA in all investigated archaeal strains . PLEL concentration and composition was estimated in accordance with previous studies with two exceptions . In the polar (phospho)lipid fraction of Methanopyrus kandleri side chains possibly derived from hydroxyarchaeol as well as acyclic and cyclic caldarchaeol were identified . In phospholipid extracts of Methanothermus fervidus the 'H-formed' caldarchaeol could not be detected . Overall, PLEL derived isoprenoids as well as PLFA enabled taxonomic differentiation of the selected microorganisms into phylogenetically related groups. Exp Cell Res, 2002 Aug 1, 278(1), 112 - 22 Spatiotemporal regulation of moesin phosphorylation and rear release by Rho and serine/threonine phosphatase during neutrophil migration; Yoshinaga-Ohara N et al.; Neutrophil motility is crucial to effective host defenses against microorganisms . While uropod retraction is a critical step in the migration of neutrophils, the underlying molecular mechanism is not well understood . Here, we show that inhibition of the Rho small GTPase with C3 exoenzyme prevented the retraction of trailing uropods, indicating that the process of rear release is mediated by a Rho signaling pathway . C3 exoenzyme caused marked elongation of directionally migrating neutrophils, suggesting an additional role for Rho in the maintenance of functional polarized cell shape . We also show that phosphorylation and dephosphorylation of the plasma membrane-actin filament cross-linker moesin are spatiotemporally controlled in migrating neutrophils . In particular, phosphorylation of moesin at threonine 558 depended on Rho activity . Videomicroscopy showed that dephosphorylation of this carboxy-terminal threonine preceded uropod retraction . Calyculin A, an inhibitor of type 1 and type 2A serine/threonine phosphatases, suppressed the moesin dephosphorylation and impaired uropod retraction in a dose-dependent manner . Cypermethrin, an inhibitor of type 2B serine/threonine phosphatase, had no such effects . The finding that Rho small GTPase and type 1/type 2A phosphatases are involved in rear release yields novel insights into the biochemical mechanisms of neutrophil migration. J Mol Biol, 2002 Jul 26, 320(5), 1147 - 56 Allosteric inhibition of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase alters the coordination of both substrates; Shumilin IA et al.; 3-Deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHPS), the first enzyme of the aromatic biosynthetic pathway in microorganisms and plants, catalyzes the aldol-like condensation of phosphoenolpyruvate and D-erythrose-4-phosphate with the formation of 3-deoxy-D-arabino-heptulosonate-7-phosphate . In Escherichia coli, there are three isoforms of DAHPS, each specifically feedback-regulated by one of the three aromatic amino acid end products . The crystal structure of the phenylalanine-regulated DAHPS from E.coli in complex with its inhibitor, L-phenylalanine, phosphoenolpyruvate, and metal cofactor, Mn(2+), has been determined to 2.8A resolution . Phe binds in a cavity formed by residues of two adjacent subunits and is located about 20A from the closest active site . A model for the mechanism of allosteric inhibition has been derived from conformational differences between the Phe-bound and previously determined Phe-free structures . Two interrelated paths of conformational changes transmit the inhibitory signal from the Phe-binding site to the active site of DAHPS . The first path involves transmission within a single subunit due to the movement of adjacent segments of the protein . The second involves alterations in the contacts between subunits . The combination of these two paths changes the conformation of one of the active site loops significantly and shifts the other slightly . This alters the interaction of DAHPS with both of its substrates . Upon binding of Phe, the enzyme loses the ability to bind D-erythrose-4-phosphate and binds phosphoenolpyruvate in a flipped orientation. Curr Infect Dis Rep, 2002 Aug, 4(4), 339 - 350 Molecular Techniques in the Diagnosis of Central Nervous System Infections; Lu HZ et al.; Development of polymerase chain reaction (PCR)-based molecular techniques has initiated a revolution in the field of diagnostic microbiology . These techniques have not only provided rapid, noninvasive detection of microorganisms that cause central nervous system (CNS) infections, but have also demonstrated that several neurologic disorders are linked to infectious agents . While PCR-based techniques are predicted to be widely used in diagnosing and monitoring CNS infections, the limitations, as well as strengths, of these techniques must be clearly understood by both clinicians and laboratory personnel to ensure proper utilization. Biochemistry (Mosc), 2002 Jun, 67(6), 672 - 5 Structure of O-specific polysaccharide from Pseudoalteromonas nigrifaciens strain KMM 161; Gorshkova RP et al.; On mild acid degradation of the lipopolysaccharide of the marine microorganism Pseudoalteromonas nigrifaciens KMM 161 an O-specific polysaccharide containing D-galactose, 2-acetamido-2-deoxy-D-glucose, 3,6-dideoxy-3-(4-hydroxybutyramido)-D-galactose, and 2-acetamido-2-deoxy-L-guluronic acid residues was obtained . From the results of Smith degradation, O-deacetylation of the polysaccharide, and NMR spectroscopy the following structure of the tetrasaccharide repeating unit of the O-specific polysaccharide was established {see reaction} . It should be noted that the same structure occurs in the antigenic polysaccharide of Pseudoalteromonas nigrifaciens KMM 158 described earlier as Alteromonas macleodii 2MM6. Protist, 2002 Jun, 153(2), 133 - 42 A simple and rapid PCR-based method to isolate complete small macronuclear minichromosomes from hypotrich ciliates: 5S rDNA and S26 ribosomal protein gene of Oxytricha (Sterkiella) nova; Callejas S et al.; Hypotrich ciliates present a macronuclear genome consisting of gene-sized instead of chromosome-sized DNA molecules . Exploiting this unique eukaryotic genome feature, we introduce, for the first time in ciliates, a rapid and easy PCR method using telomeric primers to isolate small complete macronuclear DNA molecules or minichromosomes . Two presumably abundant macronuclear DNA molecules, containing ribosomal genes, were amplified from the Oxytricha (Sterkiella) nova complete genome after using this method, and then were cloned and sequenced . The 5S rDNA sequence of O . (S.) nova is the third one reported among hypotrich ciliates; its primary and secondary structure is compared with other eukaryotic 5S rRNAs . The ribosomal protein S26 gene is the first one reported among ciliates . This "End-End-PCR" method might be useful to obtain similar gene-sized macronuclear molecules from other hypotrich ciliates, and, therefore, to increase our knowledge on ribosomal genes in these eukaryotic microorganisms. J Inorg Biochem, 2002 Jul 25, 91(1), 125 - 31 Primary structure of a Japanese lacquer tree laccase as a prototype enzyme of multicopper oxidases; Nitta K et al.; The cDNA library of the Japanese lacquer tree (Rhus vernicifera) was constructed by the reverse transcription of mRNA . A cDNA encoding laccase was amplified by PCR using primers based on the N-terminal amino acid sequences of the purified laccase and its peptide fragments formed by digestions with chymotrypsin and trypsin, and subcloned . The laccase cDNA clone contained a single, large open reading frame of 1599 nucleotides, encoding a protein of 533 amino acids with a calculated molecular mass of 58981 Da . The lacquer laccase was found to have 42 to 62% identity with other plant laccases and 20 to 24% identity with microorganism laccases at the deduced amino acid level . Differing from microorganism laccases the lacquer laccase utilizes a Met residue in addition to one Cys and two His residues to construct the type 1 Cu site . The secondary structure of the lacquer laccase was predicted to mainly consist of the beta-structure (28.7%) and loop and random structures (67.0%) . The alpha-helix content was predicted to be only 4.3% . The location of these secondary structures was assumed to be very similar to those of ascorbate oxidase and fungal laccase, the crystal structures of which have been determined. Mol Plant Microbe Interact, 2002 Jul, 15(7), 693 - 700 Induction of trehalase in Arabidopsis plants infected with the trehalose-producing pathogen Plasmodiophora brassicae; Brodmann A et al.; Various microorganisms produce the disaccharide trehalose during their symbiotic and pathogenic interactions with plants . Trehalose has strong effects on plant metabolism and growth; therefore, we became interested to study its possible role in the interaction of Arabidopsis thaliana with Plasmodiophora brassicae, the causal agent of clubroot disease . We found that trehalose accumulated strongly in the infected organs (i.e., the roots and hypocotyls) and, to a lesser extent, in the leaves and stems of infected plants . This accumulation pattern of trehalose correlated with the expression of a putative trehalose-6-phosphate synthase (EC 2.4.1.15) gene from P . brassicae, PbTPS1 . Clubroot formation also resulted in an induction of the Arabidopsis trehalase gene, ATTRE1, and in a concomitant increase in trehalase (EC 3.2.1.28) activity in the roots and hypocotyls, but not in the leaves and stems of infected plants . Thus, induction of ATTRE1 expression was probably responsible for the increased trehalase activity . Trehalase activity increased before trehalose accumulated; therefore, it is unlikely that trehalase was induced by its substrate . The induction of trehalase may be part of the plant's defense response and may prevent excess accumulation of trehalose in the plant cells, where it could interfere with the regulation of carbon metabolism. Mem Inst Oswaldo Cruz, 2002 Jun, 97(4), 443 - 57 Some aspects of protozoan infections in immunocompromised patients- a review; Ferreira MS et al.; Protozoa are among the most important pathogens that can cause infections in immunocompromised hosts . These microorganisms particularly infect individuals with impaired cellular immunity, such as those with hematological neoplasias, renal or heart transplant patients, patients using high doses of corticosteroids, and patients with acquired immunodeficiency syndrome . The protozoa that most frequently cause disease in immunocompromised patients are Toxoplasma gondii, Trypanosoma cruzi, different Leishmania species, and Cryptosporidium parvum; the first two species cause severe acute meningoencephalitis and acute myocarditis, Leishmania sp . causes mucocutaneous or visceral disease, and Cryptosporidium can lead to chronic diarrhea with hepatobiliary involvement . Various serological, parasitological, histological and molecular methods for the diagnosis of these infections are currently available and early institution of specific therapy for each of these organisms is a basic measure to reduce the morbidity and mortality associated with these infections. Infect Immun, 2002 Aug, 70(8), 4247 - 53 Leukotriene B(4) induces nitric oxide synthesis in Trypanosoma cruzi-infected murine macrophages and mediates resistance to infection; Talvani A et al.; The production of nitric oxide (NO) by gamma interferon (IFN-gamma)-activated macrophages is a major effector mechanism during experimental Trypanosoma cruzi infection . In addition to IFN-gamma, chemoattractant molecules, such as platelet-activating factor (PAF) and CC chemokines, may also activate macrophages to induce NO and mediate the killing of T . cruzi in an NO-dependent manner . Here we investigated the ability of leukotriene B(4) (LTB(4)) to induce the production of NO by macrophages infected with T . cruzi in vitro and whether NO mediated LTB(4)-induced parasite killing . The activation of T . cruzi-infected but not naive murine peritoneal macrophages with LTB(4) induced the time- and concentration-dependent production of NO . In addition, low concentrations of LTB(4) acted in synergy with IFN-gamma to induce NO production . The NO produced mediated LTB(4)-induced microbicidal activity in macrophages, as demonstrated by the inhibitory effects of an inducible NO synthase inhibitor . LTB(4)-induced NO production and parasite killing were LTB(4) receptor dependent and were partially blocked by a PAF receptor antagonist . LTB(4) also induced significant tumor necrosis factor alpha (TNF-alpha) production, and blockade of TNF-alpha suppressed LTB(4)-induced NO release and parasite killing . A blockade of LTB(4) or PAF receptors partially inhibited IFN-gamma-induced NO and TNF-alpha production but not parasite killing . Finally, daily treatment of infected mice with CP-105,696 was accompanied by a significantly higher level of blood parasitemia, but not lethality, than that seen in vehicle-treated animals . In conclusion, our results suggest a role for LTB(4) during experimental T . cruzi infection . Chemoattractant molecules such as LTB(4) not only may play a major role in leukocyte migration into sites of inflammation in vivo but also, in the event of an infection, may play a relevant role in the activation of recruited leukocytes to kill the invading microorganism in an NO-dependent manner. Med Pr, 2002, 53(2), 145 - 9 {Risk assessment of known and new chemicals}; Dawidek-Pietryka K et al.; It has become obvious that exposure to chemicals entails risk . Their hazardous and threatening effects may be not only direct but also indirect affecting microorganisms, animals and plants . This aspect of hazard and risk assessment is almost identical to that of toxicity . There are various categories of chemicals and for each of them risk assessment regulations have been developed . There are two major groups of chemicals: those which are already known and new ones . There are also some specific categories of chemical compounds, such as pesticides, biocides, medicinal products, cosmetics, food additives, feed additives, as well as radioactive substances and others . At present, the binding national and international regulations regarding different categories of chemical compounds are the subject of discussion in the member states of the European Union . In Poland, and attempt has been made to find appropriate examples of legal regulations that could be followed in our country, and this problem remains still open. Arch Environ Contam Toxicol, 2002 Aug, 43(2), 168 - 74 Suppression of pyrite oxidation by iron 8-hydroxyquinoline; Lan Y et al.; One of the important approaches to prevent pyrite (FeS(2)) oxidation and subsequent formation of acid mine drainage (AMD) is to create a surface coating on pyrite . In this study, a coating of iron 8-hydroxyquinoline was formed by leaching pyrite with a 0.10 M H(2)O(2)/0.0034 M 8-hydroxyquinoline solution; stability of the coated pyrite was tested under various pH and temperature conditions . The results showed that iron 8-hydroxyquinoline coating could significantly suppress further pyrite oxidation by both chemical (H(2)O(2)) and biological ( e.g., Thiobacillus ferrooxidans) processes . At pH from 3.0 to 5.0 and temperature from 10-40 degrees C, the amount of SO(4)(2-) leached out by 0.10 M H(2)O(2) from the coated pyrite samples was 54.8-70.1% less than that from the uncoated controls . The oxidation of pyrite followed a pseudo-zero-order kinetics under the constant concentration of H(2)O(2) . In the presence of microorganisms, sulfate leached out of the uncoated pyrite in 1 year accounted for 5.32% of the total pyrite in the system, with a concurrent pH drop to 2.35 under the ambient room temperatures . In contrast, the amount leached out from the coated samples was only 0.15% of the total pyrite and the final pH was 5.48 . Thus, the coating decreased the leachability of pyrite by 97% in the inoculated systems . In comparison to the more widely studied iron phosphate coating, the advantage of iron 8-hydroxyquinoline coating was that it inhibited both chemical and biological pyrite oxidation, whereas iron phosphate coating could only inhibit chemical pyrite oxidation. FEMS Microbiol Lett, 2002 Jul 2, 212(2), 217 - 20 Chemical stress-responsive genes from the lignin-degrading fungus Phanerochaete chrysosporium exposed to dibenzo-p-dioxin; Kurihara H et al.; The stress-responsive genes expressed against the exogenous addition of dibenzo-p-dioxin from the lignin-degrading basidiomycete, Phanerochaete chrysosporium, were determined utilizing a differential display reverse transcription-PCR technique . Six cDNA fragments, exhibiting a high homology with various proteins from other microorganisms, were identified via a BLAST search; that is, NADH-ubiquinone oxidoreductase (NUO), ATP/ADP carrier, uric acid-xanthine permease, manganese superoxide dismutase, 3-hydroxybutyryl-coenzyme A dehydrogenase, and cytoskeletal protein . The expression of NUO was also up-regulated by catechol and trihydroxybenzene but not by dibenzofuran, suggesting that NUO expression was initiated by the formation of quinone products through the reaction of extracellular one-electron oxidizing enzymes with dibenzo-p-dioxin. Chirality, 2002 Jun, 14(6), 495 - 7 Regio- and stereoselective hydroxylation of taxoids by filamentous fungi; Hu S et al.; Paclitaxel (Taxol), is one of the most promising chemotherapeutic agents developed for cancer treatment in past two decades . Microorganisms such as filamentous fungi are known to perform regio- and stereoselective hydroxylation of taxoids . Herein, we describe highly regio- and stereoselective hydroxylation at the 1beta and 9alpha positions of the taxane skeleton . Such hydroxylation reactions proceed readily for the taxadienes as substrates rather than taxoids having an oxetane ring . The presence of different oxygen substituents on the taxane nucleus, such as 5-acetoxy, has a significant effect on the selectivity and yield of the hydroxylation catalyzed by the microbial oxidases . J Mol Model (Online), 2002 Jan, 8(1), 8 - 23 Comparative molecular field analysis (CoMFA) for sulfoxidation reactions in Mortierella isabellina ATCC 42613 and Helminthosporium sp . NRRL 4671; Huang WH et al.; Previous models for mechanisms of enzymatic sulfoxidation have been somewhat limited by a lack of knowledge of the essential features of substrate-enzyme versus product-enzyme relationships . Computerized methods for modeling ligand-protein (substrate-enzyme) interactions can overcome some of these limitations . Specifically, CoMFA (comparative molecular field analysis) provided a useful general approach in which to evaluate substrate-enzyme and product-enzyme relationships . The present investigation examined the relationship between substrate and product structure in predicting enantioselective sulfoxidation reactions using CoMFA for two species of microorganisms that have been used as models for mammalian metabolism, Mortierella isabellina and Helminthosporium sp . The overall enantioselectivity observed was based on the composite stereoselectivity of sulfoxide formation, sulfone formation (from the sulfoxide), and sulfoxide reduction back to the achiral substrate (sulfide). Appl Microbiol Biotechnol, 2002 Jul, 59(2-3), 246 - 51 Epub 2002 May 04. Expression of a gene for Mn-peroxidase from Coriolus versicolor in transgenic tobacco generates potential tools for phytoremediation; Iimura Y et al.; In efforts aimed at the detoxification of contaminated areas, plants have many advantages over bacteria and fungi . We are attempting to enhance the environmental decontamination functions of plants by transferring relevant genes from microorganisms . When the gene for Mn-peroxidase (MnP) from Coriolus versicolor was expressed in transgenic tobacco plants, one line (designated fMnP21) expressed MnP activity at levels 54-fold higher than in control lines . When undamaged roots of transgenic plants were applied to liquid medium supplemented with 250 microM pentachlorophenol (PCP), the decrease in the level of PCP in fMnP21 (86% reduction) was about 2-fold higher than that in control lines (38% reduction) . Expression of the gene for MnP in the transgenic plants had no obvious negative effects on their vegetative and sexual growth . Our system should contribute to the development of novel methods for the removal of hazardous chemicals from contaminated environments using transgenic plants. EMBO J, 2002 Jul 15, 21(14), 3632 - 42 Ferrichrome induces endosome to plasma membrane cycling of the ferrichrome transporter, Arn1p, in Saccharomyces cerevisiae; Kim Y et al.; Siderophores are small iron-binding molecules that are synthesized and secreted in the iron-free form by microorganisms . Saccharomyces cerevisiae takes up iron bound to siderophores by two separate systems, one of which requires the ARN family of sidero phore-iron transporters . Arn1p and Arn3p are expressed in endosome-like intracellular vesicles . Here we present evidence that, in the absence of its specific substrate, ferrichrome, Arn1p is sorted directly from the Golgi to the endosomal compartment and does not cycle to the plasma membrane . When cells are exposed to ferrichrome at low concentrations, Arn1p stably relocalizes to the plasma membrane . At higher concentrations of ferrichrome, Arn1p relocalizes to the plasma membrane and rapidly undergoes endocytosis . Plasma membrane localization of Arn1p occurs only in the presence of its specific substrate, and not in the presence of other siderophores . Despite expression of Arn1p on the plasma membrane, mutant strains with defects in endocytosis exhibit reduced uptake of ferrichrome-iron . Thus, siderophores influence the trafficking of the Arn transporters within the cell and this trafficking is important for transporter function. PDA J Pharm Sci Technol, 2002 May-Jun, 56(3), 150 - 71 Method for qualifying microbial removal performance of 0.1 micron rated filters . Part IV: Retention of hydrogenophaga pseudoflava (ATCC 700892) and Ralstonia pickettii (ATCC 700591) by 0.2 and 0.22 micron rated filters; Sundaram S et al.; Ralstonia pickettii has emerged as a bioburden microorganism of considerable importance in pharmaceutical processes utilizing conventional 0.2 or 0.22 micron rated "sterilizing grade" filters . In this article, we re-evaluated and studied the retention efficiencies of 0.2 micron rated nylon 6.6 and 0.22 microns rated modified polyvinylidene fluoride (PVDF) filters for Hydrogenophaga pseudoflava (ATCC 700892) and R . pickettii (ATCC 700591) . Out of a total of forty-four 0.2/0.22 micron rated filters discs tested in this study (spanning different challenge fluids, different challenge conditions, and different filter types), H . pseudoflava penetration was observed for every filter disc tested . Log titer reduction (LTR) values ranged from 0.3 to 2.0 logs for 20-48 hour challenges conducted in Water for Injection (WFI), and 3.8-7.1 logs for 6-hour challenges conducted in Minimal Media Davis (MMD) . For 0.2 micron nylon 6.6 filter discs, penetration by R . pickettii was observed only in WFI challenges and was dependent on the culture and challenge conditions used . Penetration by R . pickettii was also restricted to only those membrane discs that were very close to the filter manufacturer's production integrity test (the Quantitative Bubble Point, QBP, test) limit . Where R . pickettii penetration was observed, LTR values were significantly higher than those observed for H . pseudoflava with the same filter discs . This study: 1) supports the use of H . pseudoflava as a worst-case challenge model for R . pickettii in process- and product-specific bacterial retention testing; 2) provides experimental evidence, for the first time, for the need to include filter membrane lots that have a physical integrity test value at or near the filter manufacturer's production (lower) limit in these tests; and 3) demonstrates how a standardized membrane integrity test (such as the QBP test) can be used select such "worst-case" membranes and to verify the inclusion of such "worst-case" membranes in these tests, thus serving as the link between the membrane disc used in bacterial retention validation testing and the production process filter. J Chromatogr A, 2002 May 17, 956(1-2), 261 - 70 Ion chromatographic determination of trace hydroxylamine in waste streams generated by a pharmaceutical reaction process; Fernando PN et al.; Hydroxylamine is a key raw material used in a synthetic drug process at Pharmacia . Since hydroxylamine is harmful to microorganisms, concentrations above 5 ppm could interfere with the biological sewage plant performance . This necessitated the development of a sensitive analytical method for detecting low levels of hydroxylamine in the waste streams generated from the pharmaceutical process . The present report describes a cation-exchange chromatographic method coupled with pulsed amperometric detection at a gold electrode for trace analysis of hydroxylamine . This method was evaluated by generating data on the parameters of specificity, precision, linearity, recovery and sensitivity. Proc Natl Acad Sci U S A, 2002 Jul 23, 99(15), 9727 - 32 Epub 2002 Jul 08. Trehalose synthesis is induced upon exposure of Escherichia coli to cold and is essential for viability at low temperatures; Kandror O et al.; Trehalose accumulates dramatically in microorganisms during heat shock and osmotic stress and helps protect cells against thermal injury and oxygen radicals . Here we demonstrate an important role of this sugar in cold-adaptation of bacteria . A mutant Escherichia coli strain unable to produce trehalose died much faster than the wild type at 4 degrees C . Transformation of the mutant with the otsA/otsB genes, responsible for trehalose synthesis, restored trehalose content and cell viability at 4 degrees C . After temperature downshift from 37 degrees C to 16 degrees C ("cold shock"), trehalose levels in wild-type cells increased up to 8-fold . Although this accumulation of trehalose did not influence growth at 16 degrees C, it enhanced cell viability when the temperature fell further to 4 degrees C . Before the trehalose build-up, levels of mRNA encoding OtsA/OtsB increased markedly . This induction required the varsigma factor, RpoS, but was independent of the major cold-shock protein, CspA . otsA/B mRNA was much more stable at 16 degrees C than at 37 degrees C and contained a "downstream box," characteristic of cold-inducible mRNAs . Thus, otsA/otsB induction and trehalose synthesis are activated during cold shock (as well as during heat shock) and play an important role in resistance of E . coli (and probably other organisms) to low temperatures. Circulation, 2002 Jul 9, 106(2), 184 - 90 Predisposition to atherosclerosis by infections: role of endothelial dysfunction; Prasad A et al.; BACKGROUND: Several microorganisms have been implicated in the pathogenesis of atherosclerosis . We hypothesized that infections may predispose to atherosclerosis by inflicting endothelial injury . METHODS AND RESULTS: Of 375 patients undergoing coronary angiography, 218 had assessment of endothelial function using intracoronary acetylcholine (ACH) and of endothelium-independent function with sodium nitroprusside and adenosine . Immunoglobulin-G antibody titers to cytomegalovirus, Chlamydia pneumoniae, Helicobacter pylori, hepatitis A virus, and herpes simplex virus-1 were measured . Pathogen burden was defined as the number of positive antibodies . Although positive serology to individual pathogens tended to be associated with increased incidence of coronary arteriosclerosis (CAD), the pathogen burden correlated with the presence of CAD, even after adjustment for risk factors (OR 1.3; 95% CI, 1.05 to 1.6, P=0.018) . Moreover, the severity of CAD was independently associated with the pathogen burden (P=0.001) . Pathogen burden was an independent predictor of endothelial dysfunction, determined as the percent change in coronary vascular resistance in response to ACH (P=0.009) but not the responses to sodium nitroprusside or adenosine . Pathogen burden was also an independent determinant of endothelial function in the subgroup with angiographically normal coronary arteries . CONCLUSIONS: The immunoglobulin-G antibody response to multiple pathogens (pathogen burden) is an independent risk factor for endothelial dysfunction and the presence and severity of CAD . Endothelial dysfunction provides the crucial link by which pathogens may contribute to atherogenesis. Biomol Eng, 2002 Jun, 19(1), 17 - 30 Modification of polysaccharides and plant cell wall by endo-1,4-beta-glucanase and cellulose-binding domains; Levy I et al.; Cellulose is one of the most abundant polymers in nature . Different living systems evolved simultaneously, using structurally similar proteins to synthesize and metabolize polysaccharides . In the growing plant, cell wall loosening, together with cellulose biosynthesis, enables turgor-driven cell expansion . It has been postulated that endo-1,4-beta-glucanases (EGases) play a central role in these complex activities . Similarly, microorganisms use a consortium of lytic enzymes to convert cellulose into soluble sugars . Most, if not all, cellulases have a modular structure with two or more separate independent functional domains . Binding to cellulose is mediated by a cellulose-binding domain (CBD), whereas the catalytic domain mediates hydrolysis . Today, EGases and CBDs are known to exist in a wide range of species and it is evident that both possess immense potential in modifying polysaccharide materials in-vivo and in-vitro . The hydrolytic function is utilized for polysaccharide degradation in microbial systems and cell wall biogenesis in plants . The CBDs exerts activity that can be utilized for effective degradation of crystalline cellulose, plant cell wall relaxation, expansion and cell wall biosynthesis . Applications range from modulating the architecture of individual cells to an entire organism . These genes, when expressed under specific promoters and appropriate trafficking signals can be used to alter the nutritional value and texture of agricultural crop and their final products . EGases and CBDs may also find applications in the modification of physical and chemical properties of composite materials to create new materials possessing improved properties. Bioresour Technol, 2002 Jul, 83(3), 221 - 4 Microbial detoxification of waste rubber material by wood-rotting fungi; Bredberg K et al.; The extensive use of rubber products, mainly tires, and the difficulties to recycle those products, has resulted in world wide environmental problems . Microbial devulcanisation is a promising way to increase the recycling of rubber materials . One obstacle is that several microorganisms tested for devulcanisation are sensitive to rubber additives . A way to overcome this might be to detoxify the rubber material with fungi prior to the devulcanisation . In this study, 15 species of white-rot and brown-rot fungi have been screened with regard to their capacity to degrade an aromatic model compound in the presence of ground waste tire rubber . The most effective fungus, Resinicium bicolor, was used for detoxification of rubber material . Increase in growth of the desulfurising bacterium Thiobacillus ferrooxidans in presence of the rubber treated with Resinicium bicolor compared to untreated rubber demonstrated that detoxification with fungi is possible. Adv Mar Biol, 2002, 42, 137 - 236 The Scaphopoda; Reynolds PD; The Scaphopoda are marine infaunal carnivores that feed on foraminiferans and other microorganisms selected and manipulated by their unique feeding tentacles or captacula . Their tusk-like shell is open at both ends; the burrowing foot and captacula protrude anteriorly, while respiratory currents pass through the posterior opening . Although the scaphopods comprise one of the smallest molluskan classes in terms of species diversity, they have a worldwide distribution ranging from intertidal to depths in excess of 6000 m . Despite detailed monographic work from the late nineteenth and early twentieth centuries, however, the biology of scaphopods is today among the least understood of mollusks . To some extent, this is related to a significant departure from more typical molluscan organization inferred from studies of the larger classes . For example, the mantle cavity is elongate and restricted in space, and several associated organs are lost, including the osphradium and ctenidia; the latter is associated with the loss of auricles from the scaphopod heart . There is also little record of ontogeny in the adult shell, as the older portion is periodically decollated to maintain the posterior aperture for passage of inhalant and exhalant currents . This and other constraints on scaphopod diversification are reflected in the range limits of shell shape within the class . In contrast, the "Dentalium" egg has been a model system for experimental embryology, particularly in studies of cell lineage and early morphogenesis . Later larval development and most organogenesis is nonetheless poorly known, although recent studies have considerably enhanced our knowledge in this area . Global biogeographic patterns of scaphopod diversity have been studied only preliminarily . A general decrease in diversity with depth is broken by small diversity peaks at bathyal depths . There is also evidence for a marked latitudinal diversity gradient in the world's oceans, near equatorial in the Pacific but at approximately 20 degrees N in the Atlantic . Scaphopods have a wide diversity of ecto- and endo-symbiotic associations with other organisms, including commensal bacteria and ciliates, mutualistic anemones and corals, and parasitic algae and platyhelminths . Other documented associations include predation by naticid gastropods and ratfish, and the inhabitation of empty scaphopod shells by a variety of sipunculans and hermit crabs . Phylogeny within the class is still unresolved, although significant progress has been made recently in documenting morphological variation among families, genera, and species for application in a cladistic context . The evolutionary relationships of Scaphopoda within Mollusca are farther from resolution, with every conchiferan class a proposed sister group among modern analyses . Molecular avenues should provide significant progress in scaphopod phylogenetics, as indicated by recent comparative sequence analysis and gene expression patterns . Scaphopods comprise the most recent class of mollusks to appear in the fossil record, dating at least from the Mississippian Carboniferous (362.5 My) . They appear from first-order occurrence data to have a substantially documented fossil record, with several sharp extinction and radiation events evident during their comparatively short geologic history . The unresolved position of the Scaphopoda within molluscan phylogeny, and their relatively limited range of morphological disparity but substantial derivation from other conchiferans, renders the scaphopods an enigmatic group . They represent a significant evolutionary avenue within the Mollusca, but one that is poorly examined . Nonetheless, comprehensive phylogenetic analysis and documentation of diversity within this small, monophyletic, class-level taxon, well-preserved in the fossil record, is imminently tractable, and presents a singular opportunity in the study of molluscan evolution. Planta Med, 2002 Jun, 68(6), 564 - 7 Ferulagone: a new monoterpene ester from Ferulago thirkeana essential oil; Baser KH et al.; Essential oil obtained by micro-distillation and hydrodistillation of the endemic Ferulago thirkeana (Boiss.) Boiss . (Apiaceae) was analysed by GC/MS . An unknown component isolated from the essential oil was characterized by chromato-spectral techniques (1D-, 2D-NMR, HRMS, IR and UV) as (1 S)-2,6,6-trimethyl-4-oxobicyclo{3.1.1}hept-2-enyl(2 E)-2-methylbut-2-enoate (= ferulagone or 1-angeloyloxyverbenone) (1) . Biological activities of the essential oil and the purified major component (1) were tested against various human pathogenic microorganisms resulting in moderate inhibition (62.5 - 125 microg/ml). Pediatr Pathol Mol Med, 2002 Jul-Aug, 21(4), 353 - 99 Pathogenesis and pathology of hematogenous infections of the fetus and newborn; Bittencourt AL et al.; The earlier infections occur in intrauterine life the more severe are they . When the infection develops during embryogenesis, the lesions are much more serious, sometimes causing disruptions (malformations) . At this time the organs are not completely formed and microorganisms may interfere with organogenesis to such an extent that the development of the functions necessary for viability become impaired . Infection acquired in utero may result in resorption of the embryo, abortion, stillbirth, neonatal death, intrauterine growth retardation (IUGR), or prematurity . The infected newborns commonly are apparently normal at birth but they may develop a late onset disease . Otherwise, neonates presenting symptomatology may derelop untoward sequelae. Infect Dis Clin North Am, 2002 Jun, 16(2), 339 - 61, ix Diagnostic methods current best practices and guidelines for histologic evaluation in infective endocarditis; Lepidi H et al.; Infective endocarditis (IE) often presents diagnostic and therapeutic challenges and continues to cause high morbidity and mortality . Confirmation of the diagnosis of IE is important for the purposes of epidemiologic and clinical studies and is crucial for patient management . Despite recent advances in diagnostic techniques, about 10% of IE cases remain culture-negative . Because pathological examination of cardiac valves to demonstrate vegetations and valvular inflammation remains the gold standard for the diagnosis of IE, the role of the pathologist is often decisive, especially when bacteriologists fail to isolate a microorganism or when a microorganism that has been isolated may be a contaminant . Furthermore, the pathologist may play an important role in identification of previously unknown infectious agents. Infect Dis Clin North Am, 2002 Jun, 16(2), 255 - 72, vii Progress toward a global understanding of infective endocarditis . Early lessons from the International Collaboration on Endocarditis investigation; Cabell CH et al.; In the century and a quarter since William Osler delivered his framed lectures on endocarditis substantial advancements have occurred in the understanding and treatment of this disease . This article summarizes current understanding of endocarditis in the areas of population epidemiology, host factors, microorganisms, diagnosis, and therapy . In addition, the authors discuss possible directions for investigation in the future, including a new multinational consortium, the International Collaboration on Endocarditis (ICE) . This collaboration aims to provide a mechanism to advance the understanding of endocarditis in areas difficult to study without an established network . The multinational nature of the collaboration may also permit a more global view of IE and provide opportunities for studies such as randomized trials of therapeutic treatment strategies. Nat Biotechnol, 2002 Aug, 20(8), 821 - 5 Epub 2002 Jul 01. Harnessing microbially generated power on the seafloor; Tender LM et al.; In many marine environments, a voltage gradient exists across the water sediment interface resulting from sedimentary microbial activity . Here we show that a fuel cell consisting of an anode embedded in marine sediment and a cathode in overlying seawater can use this voltage gradient to generate electrical power in situ . Fuel cells of this design generated sustained power in a boat basin carved into a salt marsh near Tuckerton, New Jersey, and in the Yaquina Bay Estuary near Newport, Oregon . Retrieval and analysis of the Tuckerton fuel cell indicates that power generation results from at least two anode reactions: oxidation of sediment sulfide (a by-product of microbial oxidation of sedimentary organic carbon) and oxidation of sedimentary organic carbon catalyzed by microorganisms colonizing the anode . These results demonstrate in real marine environments a new form of power generation that uses an immense, renewable energy reservoir (sedimentary organic carbon) and has near-immediate application. Int Dent J, 2002 Jun, 52 Suppl 3, 217 - 20 Methyl mercaptan production by periodontal bacteria; Nakano Y et al.; Oral malodour is principally caused by volatile sulphur compounds (VSC) such as hydrogen sulphide, methyl mercaptan and dimethyl sulphide . Methyl mercaptan is highly toxic, and its presence within a periodontal pocket suggests involvement in the induction and/or progression of periodontal disease . Methyl mercaptan is produced from L-methionine by L-methionine- alpha -deamino- gamma -mercaptomethane-lyase (METase) . METase catalyses the alpha,gamma-eliminating reaction of L-methionine, which results in the release of alpha-ketobutyrate, methyl mercaptan and ammonia . Although methyl mercaptan is produced by a variety of microorganisms, Porphyromonas gingivalis is considered to be the most potent producer . METases of P . gingivalis have been characterised and the genes responsible for their production, the mg/genes, have been sequenced . To ascertain the role of METase in P . gingivalis pathogenicity, a METase-deficient mutant strain (M1217) from P . gingivalis strain W83 was engineered . Only 7.7% of the mice infected with W83 survived 4 days after subcutaneous injection, whereas 36% of the mice infected with M1217 survived over the same time period . Many papers have reported the periodontal pathogenesis of VSC . It has been argued that methyl mercaptan may play a significant role in the pathogenicity of P . gingivalis. Trends Microbiol, 2002 Jun, 10(6), 254 - 7 To sialylate, or not to sialylate: that is the question; Vimr E et al.; Most oropharyngeal pathogens express sialic acid units on their surfaces, mimicking the sialyl-rich mucin layer coating epithelial cells and the glycoconjugates present on virtually all host cell surfaces and serum proteins . Unlike the host's cells, which synthesize sialic acids endogenously, several microbial pathogens use truncated sialylation pathways . How microorganisms regulate sialic acid metabolism to ensure an adequate supply of free sugar for surface remodeling is a new area of research interest to basic scientists and those focused on the clinical outcome of the host-pathogen interaction. Respir Care, 2002 Jul, 47(7), 761 - 8 Physiology of airway mucus clearance; Rubin BK; Respiratory tract secretions consist of mucus, surfactant, and periciliary fluid . The airway surface fluid is present as a bilayer, with a superficial gel or mucous layer and a layer of periciliary fluid interposed between the mucous layer and the epithelium . A thin layer of surfactant separates the mucous and periciliary fluid layers . The mucous layer extends from the intermediate airway to the upper airway and is approximately 2-10 microm thick in the trachea . Airway mucus is the secretory product of the goblet cells and the submucosal glands . It is a nonhomogeneous, adhesive, viscoelastic gel composed of water, carbohydrates, proteins, and lipids . In health, the mucous gel is primarily composed of a 3-dimensional tangled polymer network of mucous glycoproteins or mucin . Mucin macromolecules are 70-80% carbohydrate, 20% protein, and 1-2% sulfate bound to oligosaccharide side chains . The protein backbones of mucins are encoded by mucin genes (MUC genes), at least 8 of which are expressed in the respiratory tract, although MUC5AC and MUC5B are the 2 principal gel-forming mucins secreted in the airway . Mucus is transported from the lower respiratory tract into the pharynx by air flow and mucociliary clearance . Expectorated sputum is composed of lower respiratory tract secretions along with nasopharyngeal and oropharyngeal secretions, cellular debris, and microorganisms . Disruption of normal secretion or mucociliary clearance impairs pulmonary function and lung defense and increases risk of infection . When there is extensive ciliary damage and mucus hypersecretion, airflow-dependent mucus clearance such as cough becomes critically important for airway hygiene. J Dairy Sci, 2002 May, 85(5), 1183 - 90 Response of forage fiber degradation by ruminal microorganisms to branched-chain volatile fatty acids, amino acids, and dipeptides; Yang CM; This study evaluated the effect of branched-chain volatile fatty acids (VFA; isobutyric acid, isovaleric acid), amino acids (valine, leucine), and dipeptides (valine-valine, leucine-leucine) on neutral detergent fiber (NDF) degradation by rumen microorganisms in vitro . The CP (%) and in situ NDF degradation rate (%/h) for alfalfa, bermudagrass, and pangolagrass hays, and napiergrass silage were 17.2 and 7.5, 4.7 and 3.1, 8.3 and 5.3, and 9.6 and 3.4, respectively . In vitro NDF digestibility was the lowest for bermudagrass; alfalfa and napiergrass were the highest . When the incubation contained more ammonia initially, digestibilities increased, but relative differences among forages were unchanged . Adding branched-chain VFA (2 mM) to incubations increased digestibilities more than controls on 15 out of 16 occasions . The effectiveness varied with isoacids and forages used . Amino acid (2 mM) or dipeptide (1 mM) addition consistently increased digestibility over controls . Amino acids further increased digestibility over corresponding isoacids on 14 occasions . Improvement in digestibility over control by leucine appeared to be greater than that by valine . Digestibilities with dipeptides were always greater than those with isoacids, except for one case . Dipeptide addition further increased digestibility significantly over corresponding amino acids on only six occasions, while percent improvement in digestibility numerically by dipeptides occurred in 10 cases . Valine-valine seemed to exert different effect than leucine-leucine, depending on initial ammonia availability . The results indicate that dipeptides could be more effective than isoacids and amino acids in improving NDF digestion . Forages with high CP content or rapid NDF degradation rate appeared to respond to additives to smaller degrees. Cancer Metastasis Rev, 2001, 20(3-4), 245 - 77 On the role of cell surface carbohydrates and their binding proteins (lectins) in tumor metastasis; Gorelik E et al.; This review focuses on the recent advances in investigations of the role of cell surface carbohydrates in tumor metastasis . It also summarizes the results of extensive studies of endogenous lectins, their structure, carbohydrate specificity and biological functions with the major emphasis on the significance of lectin-cell surface carbohydrate interactions in a metastatic process . Numerous data demonstrate that malignant transformation is associated with various and complex alterations in the glycosylation process . Some of these changes might provide a selective advantage for tumor cells during their progression to more invasive and metastatic phenotype . Cell glycosylation depends on the expression and function of various glycosyltransferases and glycosidases . Recently, transfection of genes encoding various glysosyltransferases gene in sense and antisense orientation helped to bring direct evidence that changes in cell surface carbohydrates are important for the metastatic behavior of tumor cells . Cell surface carbohydrates affect tumor cell interactions with normal cells or with the extracellular matrix during metastatic spread and growth . These interactions can be mediated via tumor cell carbohydrates and their binding proteins known as endogenous lectins . The family of the discovered endogenous lectins is rapidly expanding . The number of C-type lectins has reached 50 and at least 10 galectins have been identified . The biological significance of the endogenous lectins and their possible role in tumor growth and metastasis formation has started to unravel . Some lectins recognize the 'foreign' patterns of cell surface carbohydrates expressed by microorganisms and tumor cells, and play a role in innate and adaptive immunity . It was shown that lectins affect tumor cell survival, adhesion to the endothelium or extracellular matrix, as well as tumor vascularization and other processes that are crucial for metastatic spread and growth. J Environ Sci Health B, 2002 Jul, 37(4), 345 - 53 Persistence and bioactivity of metsulfuron-methyl in three soils; Ismail BS et al.; The persistence of metsulfuron-methyl (methyl 2-{{(4-methoxy-6-methyl-1,3,5-triazin-2-yl)aminocarbonyl}aminosul fonyl}benzoate) in nonautoclaved and autoclaved Selangor, Lating, and Serdang series soils incubated at different temperatures and with different moisture contents was investigated under laboratory conditions using cucumber (Cucumis sativus L.) as the bioassay species . Significant degradation of metsulfuron-methyl was observed in nonautoclaved soil compared with the autoclaved soil sample, indicating the importance of microorganisms in the breakdown process . At higher temperatures the degradation rate in nonautoclaved soil improved with increasing soil moisture content . In nonautoclaved Selangor, Lating and Serdang series soils, the half-life was reduced from 4.79 to 2.78 days, 4.9 to 3.5, and from 3.3 to 1.9 days, respectively, when the temperature was increased from 20 degrees to 30 degrees C at 80% field capacity . Similarly, in nonautoclaved soil, the half-life decreased with an increasing soil moisture from 20% to 80% at 30 degrees C in the three soils studied . In the autoclaved soil, the half-life values were slightly higher than those obtained in the nonautoclaved soils, perhaps indicating that the compound may be broken down by nonbiological processes . The fresh weight of the bioassay species was reduced significantly in Serdang series soil treated with metsulfuron-methyl at 0.1 ppm . However, the reduction in fresh weight of the seedlings was least in Lating series soil, followed by Selangor series soil. Chem Biol, 2002 Jun, 9(6), 721 - 9 Engineering deoxysugar biosynthetic pathways from antibiotic-producing microorganisms . A tool to produce novel glycosylated bioactive compounds; Rodriguez L et al.; A plasmid (pLN2) was generated in which genes involved in the biosynthesis of L-oleandrose in the oleandomycin producer Streptomyces antibioticus ATCC11891 were cloned . pLN2 was used to direct the biosynthesis of different deoxysugars by exchanging and/or adding genes from other antibiotic biosynthetic clusters . Transfer of the synthesized deoxysugars to the tetracenomycin C aglycon, 8-demethyl-tetracenomycin C, through the use of the "sugar flexible" glycosyltransferase ElmGT, validated the system . Several pLN2 derivatives were constructed by replacement of the oleU 4-ketoreductase gene by different 4-ketoreductase genes . Some of them, such as EryBIV and UrdR, reduced the keto group of the 4-keto intermediates, generating L-olivosyl and D-olivosyl derivatives, respectively . The system was also used to generate an L-rhamnosyl derivative (through a two-gene deletion) and an L-rhodinosyl derivative (through a combination of a gene replacement and a gene addition). Rinsho Byori, 2002 May, 50(5), 474 - 8 {Laboratory tests for infectious diseases and medical reimbursement}; Shimetani N; Emerging and reemerging infectious diseases, drug-resistant bacteria and nosocomial infection are becoming increasingly serious social problems . To control infectious diseases, it is important to develop effective therapies and to achieve complete prevention of pathogen transmission . For these purposes, rapid detection of pathogenic microorganisms is essential . Recent development and technical innovation in laboratory tests and diagnosis for infectious diseases are remarkable . New techniques such as antigen detection and genetic diagnosis, neither of which require conventional culture, have been intensively developed and introduced into clinical practice . These methods are now widely accepted as effective diagnostic tools because of their high specificity, high sensitivity and rapidity . In this article, we describe the burden of medical expenses under the law on novel infectious diseases and medical reimbursement for diagnostic tests for these diseases. Bioorg Khim, 2002 May-Jun, 28(3), 196 - 208 {Bromo- and iodo-containing alkaloids from marine microorganisms and sponges}; Dembitsky VM; The taxonomic distribution, structure, and biological activity of halogenated alkaloids isolated from marine microorganisms and sponges are reviewed . The structures of nearly 140 natural bromo- and iodo-containing alkaloids are shown. Int J Food Microbiol, 2002 Jul 25, 77(1-2), 71 - 81 Alicyclobacillus acidoterrestris spores as a target for Cupuaçu (Theobroma grandiflorum) nectar thermal processing: kinetic parameters and experimental methods; Vieira MC et al.; The kinetic parameters of thermal inactivation of a spore former, Alicyclobacillus acidoterrestris, in a tropical fruit nectar {25% of Cupuacu (Theobroma grandiflorum) pulp and 15% sugar} were determined by the isothermal method (IM), under batch heating, and by the paired equivalent isothermal exposures (PEIE) method, under non-isothermal continuous conditions . The isothermal experiments were repeated three times, every 4 months, with the same spore suspension kept frozen between experiments . The aging of spores, under frozen storage, seemed to produce a notorious increase in the z-value from experiment to experiment: Experiment 1 (z = 7.8 +/- 2.6 degrees C, D(95 degrees C) = 5.29 +/- 0.96 min), Experiment 2 (z = 22 +/- 5 degrees C, D(95 degrees C) = 5.99 +/- 0.63 min), and Experiment 3 (z =29 +/- 10 degrees C, D(95 degrees C) = 3.82 +/- 0.48 min) . The evaluation of the kinetic parameters by the PEIE method was carried out in parallel with Experiment 3, with the same aged spores, and the results (z = 31 +/- 6 degrees C, D(95 degrees C) = 5.5 +/- 1.2 min) were close to the ones obtained in this experiment . From this work, it seems that the PEIE method can also be applied to evaluate the reduction parameters of a spore-forming microorganism, and in a more realistic way, since the continuous system eliminates the errors caused by come-up and cool-down times (CUT and CDT) that are unavoidable in isothermal experiments . Therefore, when designing a thermal process for a continuous system, the PEIE method should be used, or the chances are that the process would be underdesigned, risking that the desired level of spore inactivation would not be achieved . An optimization of the thermal processing conditions was next performed for Cupuacu nectar, considering a 5D reduction in A . acidoterrestris spores . If a pasteurization process is considered, the conditions that ensure safety (9 min at 98 degrees C) only allow a 55% retention of ascorbic acid (AA) . If sterilization is considered, 8 s at 115 degrees C will ensure a safe product and retain 98.5% of the original ascorbic acid . Therefore, if A . acidoterrestris is considered as the target microorganism, the nectar should undergo an aseptic high temperature short time principle (HTST) process to achieve a 5D reduction in this acidophilus spore former . However, if the hot-fill-and-hold pasteurization process is preferred, the product should be fortified with ascorbic acid. Anaesth Intensive Care, 2002 Jun, 30(3), 275 - 82 Most nosocomial pneumonias are not due to nosocomial bacteria in ventilated patients . Evaluation of the accuracy of the 48 h time cut-off using carriage as the gold standard; Silvestri L et al.; A prospective observational cohort study was undertaken with two endpoints: (1) to compare the time cut-off of 48h and the carrier state criterion for classifying lower airway infections in adult and paediatric long-term ventilated patients, and (2) to evaluate the potential of optimized time cut-offs for characterizing imported and ICU-acquired lower airway infections . All patients admitted to the general and paediatric intensive care units and expected to require mechanical ventilation for a period > or = 3 days were enrolled . Surveillance cultures of throat and rectum were obtained on admission and thereafter twice weekly to distinguish micro-organisms that were imported into the unit from those acquired during the stay on the unit . A total of 130 adults and 400 children were studied . In the adult population, 70% of lower airway infections were classified as ICU-acquired by the 48 h cut-off and 48% by the criterion of carriage; on the paediatric ICU the percentages were 65% and 20%, respectively . To separate imported from ICU-acquired infections, eight days was optimal in the adult population and 10 days in the paediatric population . Sensitivity, specificity, positive predictive value and negative predictive value for a time cut-off of eight days for adults were 86, 77, 80, 83%, respectively, and using 10 days for children were 87, 62, 90, 56%, respectively . The use of the 48 h cut-off rule classifies patients as having nosocomial pneumonia, when in fact the infections are commonly caused by microorganisms carried in by the patients . In contrast, using the carriage method, the proportion of lung infections due to nosocomial bacteria was relatively small and was a late phenomenon . Although in prolonging the time cut-off the difference between the two types of classification was shorter, time cut-offs were still found to be unreliable for distinguishing imported from unit-acquired lower airway infections. J Ind Microbiol Biotechnol, 2002 Feb, 28(2), 70 - 3 Transformation and mineralization of benzo{a}pyrene by microbial cultures enriched on mixtures of three- and four-ring polycyclic aromatic hydrocarbons; Dries J et al.; Microorganisms originating from a soil contaminated by low levels of polycyclic aromatic hydrocarbons (PAHs) were enriched with three- and four-ring PAHs as primary substrates in the presence of benzo{a}pyrene (BaP) . Most enrichment cultures, isolated in the presence or absence of a sorptive matrix, significantly transformed BaP . Evidence of BaP mineralization was obtained with cultures enriched on phenanthrene and anthracene . Our findings supplement literature data suggesting the wide occurrence of microbial activity against BaP. Acta Microbiol Immunol Hung, 2002, 49(1), 1 - 14 Ecophysiology and breeding of mycoparasitic Trichoderma strains (a review); Manczinger L et al.; Losses due to plant diseases may be as high as 10-20% of the total worldwide food production every year, resulting in economic losses amounting to many billions of dollars and diminished food supplies . Chemical control involves the use of chemical pesticides to eradicate or reduce the populations of pathogens or to protect the plants from infection by pathogens . For some diseases chemical control is very effective, but it is often non-specific in its effects, killing beneficial organisms as well as pathogens, and it may have undesirable health, safety, and environmental risks . Biological control involves the use of one or more biological organisms to control the pathogens or diseases . Biological control is more specialized and uses specific microorganisms that attack or interfere with the pathogens . The members of the genus Trichoderma are very promising against soil-born plant parasitic fungi . These filamentous fungi are very widespread in nature, with high population densities in soils and plant litters {1} . They are saprophytic, quickly growing and easy to culture and they can produce large amounts of conidia with long lifetime. Kansenshogaku Zasshi, 2002 May, 76(5), 341 - 6 {A multicenter study of a new Helicobacter pylori selective medium . Columbia horse blood agar HP}; Hasegawa M et al.; We conducted a study for the growth of and selectivity for the desired microorganisms using a newly developed selective culture medium for Helicobacter pylori, Columbia horse blood agar HP (CHBHP), at three different Japanese clinical laboratories, Hokkaido, Kanto and Kyusyu . When standard strains and clinical isolates of H . pylori were examined, the recovery of the organism on the CHBHP media was comparable to that of conventional selective and nonselective media . However, colonies were obviously larger on the CHBHP media . These media yielded the highest H . pylori positive rate for clinical specimens at all the three laboratories . The detection rate of the CHBHP media in H . pylori-positive specimens was higher than that of media commonly used at the three laboratories (98.1% to 100% vs . 88.0% to 96.2%) . The CHBHP media also achieved a higher detection rate for specimens from H . pylori-infected animals . CHBHP media have an excellent growth supporting ability and selectivity originating from Columbia agar base and do not require the combined use of non-selective media for the growth and isolation of the organism, resulting in lower cost . Thus, they are useful media for the selective culture and isolation of H . pylori from clinical and animal specimens. Appl Microbiol Biotechnol, 2002 Jun, 59(1), 15 - 32 Epub 2002 Apr 20. Bacterial alkaline proteases: molecular approaches and industrial applications; Gupta R et al.; Proteolytic enzymes are ubiquitous in occurrence, being found in all living organisms, and are essential for cell growth and differentiation . The extracellular proteases are of commercial value and find multiple applications in various industrial sectors . Although there are many microbial sources available for producing proteases, only a few are recognized as commercial producers . A good number of bacterial alkaline proteases are commercially available, such as subtilisin Carlsberg, subtilisin BPN' and Savinase, with their major application as detergent enzymes . However, mutations have led to newer protease preparations with improved catalytic efficiency and better stability towards temperature, oxidizing agents and changing wash conditions . Many newer preparations, such as Durazym, Maxapem and Purafect, have been produced, using techniques of site-directed mutagenesis and/or random mutagenesis . Directed evolution has also paved the way to a great variety of subtilisin variants with better specificities and stability . Molecular imprinting through conditional lyophilization is coming up to match molecular approaches in protein engineering . There are many possibilities for modifying biocatalysts through molecular approaches . However, the search for microbial sources of novel alkaline proteases in natural diversity through the "metagenome" approach is targeting a hitherto undiscovered wealth of molecular diversity . This fascinating development will allow the biotechnological exploitation of uncultured microorganisms, which by far outnumber the species accessible by cultivation, regardless of the habitat . In this review, we discuss the types and sources of proteases, protease yield-improvement methods, the use of new methods for developing novel proteases and applications of alkaline proteases in industrial sectors, with an overview on the use of alkaline proteases in the detergent industry. Eur J Clin Microbiol Infect Dis, 2002 Apr, 21(4), 307 - 9 Epub 2002 Apr 13. Serological evidence of acute infection with the Chlamydia-like microorganism Simkania negevensis (Z) in acute exacerbation of chronic obstructive pulmonary disease; Lieberman D et al.; The aims of this study were twofold: (i) to test for possible associations between serological evidence of acute Simkania negevensis (Sn) infection and acute exacerbation of chronic obstructive pulmonary disease and (ii) to examine the prevalence of past infections with Sn in patients with chronic obstructive pulmonary disease . In 120 patients (63%) there was serological evidence of past infection with Sn, which was not significantly different from the rate in a control population . In five hospitalizations serological evidence existed of acute infection with Sn around the time of the exacerbation of chronic obstructive pulmonary disease . In four of these cases, there was serological evidence of acute infection with at least one other respiratory pathogen . It is concluded that Sn can be associated serologically with exacerbation of chronic obstructive pulmonary disease, in most cases together with other respiratory pathogens . The implications of these findings should be investigated further. Eur J Clin Microbiol Infect Dis, 2002 May, 21(5), 362 - 7 Epub 2002 May 07. Usefulness of sputum culture for diagnosis of bacterial pneumonia in HIV-infected patients; Cordero E et al.; The use of sputum culture in immunocompetent patients with community-acquired pneumonia is controversial . The usefulness of this technique in HIV-infected patients has not been evaluated . A prospective, observational, multicenter, hospital-based study of bacterial community-acquired pneumonia was carried out to analyze the value of sputum culture in HIV-infected patients . Only good-quality sputum samples were cultured . Altogether, 355 cases of bacterial community-acquired pneumonia were included . An etiological diagnosis was obtained in 190 (53.5%) cases . Sputum was cultured in 313 (88.1%) cases, being diagnostic in 108 (34.5%) . The microorganism identified in sputum culture was the same as that identified in sterile samples in 26 of 27 (96.3%) cases in which both cultures were diagnostic . The microbiologic findings in sputum and bronchoscopic cultures were concordant in seven of eight (87.5%) cases in which both were positive . These results suggest that sputum culture is a useful technique, given its availability and ease of performance and its good correlation with culture of sterile samples. Cornea, 2002 Jul, 21(5), 532 - 4 Fungal keratitis after nonpenetrating glaucoma surgery; Tamcelik N et al.; PURPOSE: The purpose was to report a case of fungal keratitis that occurred following viscocanalostomy . METHODS: A 63-year-old man who underwent viscocanalostomy in his left eye presented with pain, redness, watering, and a decrease in visual acuity . Slit lamp examination showed teardrop-like stromal infiltration of the superior cornea . Corneal and conjunctival scrapings were obtained and a corneal biopsy was performed . RESULTS: Microscopic examination of smears demonstrated no bacteria and fungi . Corneal biopsy revealed dichotomously branching, septate hyphae suggestive of Aspergillus species . Culture identified no microorganism . The patient responded to amphotericin B treatment and the corneal infiltration resolved, leaving a plaquelike corneal scar . CONCLUSION: Fungal keratitis may occur after viscocanalostomy, but prompt diagnosis and treatment can preserve the eye. Vet Immunol Immunopathol, 2002 Sep 10, 87(3-4), 443 - 50 The concept of sickness behavior: a brief chronological account of four key discoveries; Johnson RW; Neurons do not have receptors to detect bacteria or viruses, yet the presence of these microorganisms can cause a sickness behavior syndrome that includes, e.g., fever, anorexia, and lethargy . Because the immune system has receptors capable of detecting these non-cognitive stimuli, how the immune system transmits a message to the brain has been studied to understand why behavior is altered in sick animals . The focus has been on several cytokines secreted by leukocytes; these include interleukin (IL)-1beta, IL-6, and tumor necrosis factor-alpha (TNF-alpha) . These cytokines are secreted by activated mononuclear phagocytic cells, and numerous studies show that both peripheral and central injection of IL-1beta, IL-6, and TNF-alpha induce sickness behavior . Moreover, these cytokines and their receptors are present in the brain and inhibiting the secretion of cytokines or blocking their receptors in the brain blocks or abrogate the behavioral responses induced by inflammatory stimuli . Because the sickness behavior syndrome modulates the immune system and enhances recovery, the interplay between the immune system and central nervous system is an essential part of the overall host defense against pathogenic microorganisms . The purpose of this brief paper is to provide a chronological description of four critical advances that have led to the current understanding of how the immune system communicates with the brain to induce sickness behavior . Results from several key studies will be discussed, which showed that: (1) sickness behavior is a motivational state; (2) sickness behavior is a well-organized adaptive response to infection; (3) cytokines produced by activated leukocytes induce sickness behavior; and (4) cytokines transmit messages from the periphery to the brain using humoral and neural pathways. Vet Immunol Immunopathol, 2002 Sep 10, 87(3-4), 195 - 8 Antigen delivery systems and immunostimulation; Schijns VE; The immune system evolved to free the host from invading noxious pathogens . Vaccines are inoculated as a prophylactic measure in order to program the immune system for accelerated recognition and elimination of specific pathogens . During vaccination the immune system is exposed to attenuated or inactivated microorganisms, or their fragments . The immune response to these structures, in contrast to virulent pathogens, is often inadequate for the generation of memory cells or immune effector elements such as antibodies, perforines, granzymes or cytokines . Vaccine adjuvants help to overcome these limited responses . They provide instructive signals for the host immune system by mimicking the conditions associated with virulent infection . Hence, they either enhance and prolong expression of antigen components to reactive T cells in lymph nodes (signal 1) or they increase expression of membrane-bound or soluble costimulatory molecules (signal 2) . The enhancement of both signals by vaccine adjuvants is not mutually exclusive . Moreover, adjuvants may encode a third signal instructing the type of immune reaction to be generated . Supported by animations this presentation addresses putative immunological concepts of vaccine adjuvant activity, a phenomenon long been known as "the immunologist's dirty little secret" . Insight in the mechanisms that underlie adjuvant-induced immunostimulation and generation of memory cells will facilitate rational vaccine design. Clin Exp Dermatol, 2002 May, 27(3), 235 - 9 Associations between human leukocyte antigens and leprosy in the Turkish population; Kocak M et al.; Leprosy is a chronic infection caused by an intracellular microorganism . Genetic predisposition to both disease susceptibility and to host immunological response has been postulated for many years . The aim of this study was to determine whether there is HLA-linked susceptibility to leprosy and its different types . HLA-class I (A, B, C) and II (DR, DQ) antigen frequencies in 80 patients with leprosy (35 borderline lepromatous, 25 lepromatous, 15 borderline tuberculoid, five tuberculoid) were compared with those in 120 healthy individuals . HLA-class I antigens A9, A10, A32, B5, B21, Bw4, Bw6, Cw1, Cw2 and HLA-class II antigens DR9, DR10, DRw52, DQ1, DQ3 were found to be significantly more frequent in patients with leprosy, whereas HLA-class I antigens A3, B44, B49 and HLA-class II antigen DQ5 were so in controls . However, there was no significant difference in HLA-class I and II antigen frequencies between subtypes of leprosy . HLA-A null antigen was found to have weak expression in patients with leprosy . In conclusion, factors other than HLA-class I and class II antigens may have a more critical role in the pathophysiology of leprosy infection in man. Eur J Biochem, 2002 Jun, 269(12), 2971 - 9 EPR characterization of the mononuclear Cu-containing Aspergillus japonicus quercetin 2,3-dioxygenase reveals dramatic changes upon anaerobic binding of substrates; Kooter IM et al.; Quercetin 2,3-dioxygenase (2,3QD) is a copper-containing dioxygenase that catalyses the oxidation of the flavonol quercetin to 2-protocatechuoylphloroglucinol carboxylic acid with concomitant production of carbon monoxide . In contrast to iron dioxygenases, very little is known about copper dioxygenases . We have characterized 2,3QD from the fungus Aspergillus japonicus by electron paramagnetic resonance spectroscopy (EPR) . At pH 6.0, 2,3QD shows a mixture of two EPR species . The major form has parameters typical of type 2 Cu sites (g// = 2.330, A// = 13.7 mT), the minor one has a more distorted geometry (g// = 2.290, A// = 12.5 mT) . Anaerobic addition of the substrate quercetin results in a different, single species EPR spectrum with g// = 2.336, A// = 11.4 mT, parameters, which are in-between those of the type 2 and type 1 Cu sites in the Peisach-Blumberg (g// vs . A//) plot . After turnover, a new EPR signal is observed, which is ascribed to the carboxylic acid ester product complex . This spectrum is similar to that of the native enzyme at pH 10.0 and has g-tensor parameters suggesting a trigonal bipyramidal site . Of a variety of flavonoids studied, only flavonols are able to bind to the copper centre of 2,3QD . Nine flavonols with different hydroxylation patterns at the A- and B-ring have been analysed . They cluster in two different regions of the Peisach-Blumberg plot and show that the presence of a 5-OH group has a large effect on the A// parameter . Several differences are noted between A . japonicus 2,3QD and the enzyme from A . niger German Collection of Microorganisms 821. J Am Podiatr Med Assoc, 2002 Jun, 92(6), 327 - 30 Study of clinically suspected onychomycosis in a podiatric population; Jennings MB et al.; Onychomycosis, by definition, is a mycotic infection of the keratinized tissue of the nail plate . Although it is commonly considered to be caused by one of the dermatotropic fungi, a variety of other organisms have been implicated as etiologic agents in the disease, including some bacteria and yeasts . When it is caused by a fungus, any or all of three types of organisms can be involved: dermatophytes, yeasts, and nondermatophyte organisms . The purpose of this study was to identify the microorganisms found in fungal cultures of clinically suspected onychomycosis in the patient population of the Foot Clinics of New York in New York City, the largest foot clinic in the world . Of the 1,800 medical charts reviewed, 214 had culture results, of which 120 were positive . Trichophyton rubrum was the most prevalent pathogen, found in 67% of positive cultures . The most remarkable risk factor was age, with 80% of affected individuals older than 35 years . False-negatives may account for the high percentage (44%) of negative cultures in this study. J Microbiol Methods, 2002 Sep, 51(1), 1 - 18 Methods for detection and visualization of intracellular polymers stored by polyphosphate-accumulating microorganisms; Serafim LS et al.; Polyphosphate-accumulating microorganisms (PAOs) are important in enhanced biological phosphorus (P) removal . Considerable effort has been devoted to understanding the biochemical nature of enhanced biological phosphorus removal (EBPR) and it has been shown that intracellular polymer storage plays an important role in PAO's metabolism . The storage capacity of PAOs gives them a competitive advantage over other microorganisms present that are not able to accumulate internal reserves . Intracellular polymers stored by PAOs include polyphosphate (poly-P), polyhydroxyalkanoates (PHAs) and glycogen . Staining procedures for qualitative visualization of polymers by optical microscopy and combinations of these procedures with molecular tools for in situ identification are described here . The strengths and weaknesses of widely used polymer quantification methods that require destruction of samples, are also discussed . Finally, the potential of in vivo nuclear magnetic resonance (NMR) spectroscopy for on-line measurement of intracellular reserves is reported. Adv Ther, 2002 Mar-Apr, 19(2), 85 - 90 Phosphorylation of Helicobacter pylori CagA in patients with gastric ulcer and gastritis; Cheng KS et al.; The effects of Helicobacter pylori infection are strongly associated with chronic gastritis, gastric and duodenal ulcer, gastric cancer, and MALT lymphoma . The microorganism has been classified as a type I carcinogen by the World Health Organization . Varying clinical results from H . pylori infection are believed due, in part, to differences in virulence among species . The cag pathogenicity island is a complex of virulent genes and a coding region for the type IV phosphorylated secretion system . Through this system, many virulent gene products or proteins are phosphorylated into the host cells . This study demonstrated the positive CagA-phosphorylation effect of H . pylori in patients with chronic gastritis and benign gastric ulcer and revealed significantly different rates of CagA phosphorylation between these two diseases (P<.05). Anal Chem, 2002 Jun 1, 74(11), 2529 - 34 Mass spectrometry-based proteolytic mapping for rapid virus identification; Yao ZP et al.; A novel method is proposed for rapid identification of viruses and other organisms that show a low number of biomarkers, based on the construction of databases of organism-specific tryptic peptide masses . The peptide products of any protease that cuts at specific residues can be accommodated . Experimentally, a sample of intact virus, e.g., one collected from the atmosphere, is digested with a selective protease for a short time, and the digestion products are analyzed by MALDI-TOF mass spectrometry without fractionation or purification . In the present proof of concept, the Sindbis virus AR 339 was identified by using the masses of observed tryptic peptide products to query a database composed of tryptic peptide masses generated in silico for six viruses whose genomes have been sequenced . Two algorithms were tested for identification--a direct score-ranking algorithm and an algorithm that evaluates the probability of random matching . The Sindbis virus was unambiguously identified by either approach . The influence of factors such as experimental mass accuracy, number of missed cleavages, and database size on the identification algorithms has also been evaluated, with the objective of extending the approach to other microorganisms. Arq Gastroenterol, 2001 Oct-Dec, 38(4), 276 - 80 {Esophageal manometry: equipment cleaning and disinfection with glutaraldehyde}; Muller S et al.; BACKGROUND: Many publications have emphasized the need of proper cleaning, disinfection and sterilization process for reused materials intended to prevent cross infections . As the endoscope the esophageal manometry catheters are considered as semicritical materials and must be free of microrganisms . AIM: To standardize the esophageal manometry materials cleaning and disinfection process to guarantee the safety of patients when reusing semicritical materials . It was based on international protocols and according to recommendations of the Hospital Infection Control Commission of the "Hospital de Clinicas de Porto Alegre", Porto Alegre, RS, Brazil . MATERIALS AND METHODS: Enzymatic detergent was used for catheter cleaning, followed by immersion with 2% glutaraldehyde solution during 20 minutes for high-level disinfection . The water reservatory was kept clean and dry to prevent microorganisms proliferation . CONCLUSIONS: The high level disinfection with 2% glutaraldehyde, preceded by enzymatic detergent cleaning, is a safe and simple technique that avoids cross infection in the esophageal manometry equipment . This care must be taken after each manometric procedure . The transducers must be resterilized in ethylene oxide . The professionals of this area must work in concordance with the Hospital Infection Control Commission, being acquainted with the country laws and regulations and keeping sterilizing process and materials updated. Arq Gastroenterol, 2001 Oct-Dec, 38(4), 240 - 6 Detection of Helicobacter pylori in gastric cancer; Pereira LP et al.; BACKGROUND AND OBJECTIVES: Considering the high prevalence of stomach cancer in the northern region of Brazil and the recognized relationship between chronic gastric inflammation caused by Helicobacter pylori, and its carcinogenic potential, the objective we had with this study was to investigate the presence of the microorganism in macro and microscopic presentations of neoplasm in different regions of the stomach, and in non-malignant lesions concomitant to the adenocarcinoma in patients originating from the metropolitan area of Belem (State of Para, Brazil) . METHODS: Examinations were made on 172 patients divided into two groups: group I, formed by 75 patients with gastric carcinoma, and group II, formed by 97 patients with mild enanthematic gastritis, considered control group . The diagnosis was obtained during endoscopic examination and the respective biopsy . Gastric neoplasms were classified macroscopically in accordance with Borrmann's classification, and microscopically in accordance with Lauren's classification . In group I, 54 patients were male and 21 female while in group II, 22 patients were male and 75 female . The average age in group I was 61.2 years (range 27 to 86 years), while in group II it was 37.5 years (range 16 to 69 years) . Thin sections were prepared and stained using the hematoxylin-eosin method . In the Helicobacter pylori research, the modified Gram stain was utilized . Statistical analysis was done by utilizing the chi-squared (chi 2) test, Mann-Whitney test (U), and Fisher's exact test . RESULTS: The results showed the detection of Helicobacter pylori were significantly greater in patients with mild enanthematic gastritis than in patients with gastric carcinoma . The presence of Helicobacter pylori in patients with gastric carcinoma and mild enanthematic gastritis was significantly greater in the antral region than in other gastric regions . Helicobacter pylori detection in patients with gastric carcinoma did not present a significant difference in relation to the macroscopic aspect of the tumor either intestinal or diffuse histological types . CONCLUSIONS: These data suggest the presence of the bacteria is predominant in the antral region and it does not show relation with the macroscopic types or histological intestinal or diffuse types of gastric carcinoma. Microbes Infect, 2002 Jun, 4(7), 741 - 50 The multitalented pore-forming proteins of intracellular pathogens; Almeida-Campos FR et al.; Being an intracellular pathogen demands being able to invade a host cell, to circumvent the host immune response and to survive in the intracellular environment . Pore-forming proteins are among the innumerable tools used by intracellular microorganisms to achieve these goals . Remarkably, this seems to be a multipurpose group of proteins that can act in several ways . Making channels may signify entering into host cells, inhibiting phagocytosis, escaping phagosomes or promoting pathogen dissemination . In certain cases, pore-forming proteins are double-edged tools and may benefit the host by eliminating infected cells and/or inducing inflammation. Ageing Res Rev, 2002 Jun, 1(3), 425 - 42 Genes, mitochondria and aging in filamentous fungi; Osiewacz HD; Fungi are eukaryotic microorganisms studied in various areas of general and applied biology . A few species were among the first systems in which specific aspects of aging were addressed experimentally . Various factors, both environmental and genetic, were found to affect lifespan and aging . Mitochondrial pathways play a paramount role . Since mitochondria are semiautonomous organelles and depend on both nuclear as well as mitochondrial genes, mitochondrial-nuclear interactions are of major relevance . As a main generator of reactive oxygen species (ROS), mitochondria are prone to molecular damage . However, cells can cope with the negative effects of ROS utilizing different scavenging systems and, once defects became manifested, by repair of damaged molecules . Both, lowering ROS generation and increasing mitochondrial "caretaker" systems bear great potential to interfere with natural aging processes. J Appl Microbiol, 2002, 93(1), 69 - 76 Escherichia coli survival in groundwater and effluent measured using a combination of propidium iodide and the green fluorescent protein; Banning N et al.; AIMS: The aim of this study was to deterimine the survival of an enteric bacterium in anaerobic groundwater and effluent microcosms using the green fluorescent protein (GFP) marker gene in combination with the viability indicator propidium iodide (PI) . METHODS AND RESULTS: The pEGFP vector (Clontech) was transformed into Escherichia coli DH5alpha and was stable for at least 100 generations of growth in nonselective medium at 28 degrees C and 37 degrees C . Using an epifluorescent microscope, GFP cells could be detected under blue light (450-490 nm) and the numbers of PI-positive GFPs could be detected under green light (530-560 nm) . GFP-tagged E . coli could be detected for at least 132 d in sterilized water microcosms . GFP fluorescence was not lost from the culturable cell population for the duration of the experiment . However, a slow decline in the number of GFP-fluorescent cells in sterilized groundwater was observed . Escherichia coli die-off and loss of green fluorescence was more rapid in nonsterilized waters than in sterilized . Viable numbers of the GFP-tagged E . coli determined by PI counterstaining were compatible with numbers of colony-forming units . CONCLUSIONS: The long-term survival of E . coli and maintainance of GFP-conferred fluorescence in these cells was demonstrated in both groundwater and effluent, under sterilized conditions . However, severe starvation and/or the presence of indigenous microorganisms were found to be factors affecting the maintenance of fluorescence in dead or dying cells . SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the successful application of PI with GFP-tagging to monitor long-term bacterial survival in nutrient-limited conditions and mixed microbial populations. Caring, 2002 Jun, 21(6), 34 - 6 Maintaining skin integrity; Bryant R; One of the most basic needs of patients is to maintain intact, healthy, moisturized skin . Intact skin is the body's first line of defense against the invasion of microorganisms, provides a protective barrier from numerous environmental threats, and facilitates retention of moisture . Unfortunately, many routine patient-care activities may have a detrimental effect on the skin and consequently, they may develop into significant problems. Scand J Infect Dis, 2002, 34(4), 307 - 9 An 8-y-old boy with acute stroke and radiological signs of cerebral vasculitis after recent Mycoplasma pneumoniae infection; Ovetchkine P et al.; A young boy was admitted to hospital for acute stroke . Cerebral angiography showed a pattern suggestive of vasculitis and a recent Mycoplasma pneumoniae infection was detected . The absence of microorganisms in cerebrospinal fluid, the pattern of vasculitis and previous reports supporting a post-infectious immunological mechanism for certain complications of M . pneumoniae infection suggest this mechanism as the cause of the stroke. Hautarzt, 2002 May, 53(5), 334 - 7 {Buruli ulcer . A mycobacterial skin disease}; Thomssen H; Buruli ulcer is a chronic ulcerative skin disease caused by Mycobacterium ulcerans . It is the third most common mycobacterial disease in immunocompetent people and affects mainly children living in humid areas of the tropical rain forest . The mode of transmission is unknown . The microorganisms penetrate the skin via microinjuries . A few weeks after infection, a subcutaneous nodule develops, followed by necrosis of the subcutaneous fat and finally by a large dermal ulceration . Typical is the lack of an acute inflammatory response, likely due to an immunosuppressive toxin produced by M . ulcerans called mycolactone . The lesions mostly affect the limbs . Constitutional symptoms are normally absent . The only effective treatment consists of wide excision, often followed by skin grafts . Conservative measures are rarely successful . Buruli ulcer is characterized by low mortality and high morbidity . Early recognition and treatment are decisive for the complete cure and prevention of debilitating deformities. Z Kardiol, 2002 Apr, 91(4), 290 - 6 {Chlamydia pneumoniae and cytomegalovirus in degenerative aortic valve stenoses}; Skowasch D et al.; Recent evidence suggests a causal relationship between inflammatory as well as infectious pathomechanisms and valvular degeneration . Based on the concept of chronic Chlamydia pneumoniae and cytomegalovirus (CMV) infections, and of variable stressors working on valvular microecology, the present study sought to assess the presence of the specific chlamydial heat shock protein (cHSP) 60, of CMV, of macrophages and of the human homologue hHSP60 . Serial sections of high-grade degenerated native (n = 16) and prosthetic (n = 6) aortic valves were analyzed by immunohistochemistry for the presence of these determinants . Degenerated aortic valves revealed prevalence of Chlamydia pneumoniae in 41% (10 of 22) and CMV in 73% (16 of 22), while immunoreactive hHSP60 was present in 64% (14 of 22) and CD68 in 86% (19 of 22) . Chlamydial HSP60, CMV and hHSP60 were predominantly found in valvular fibrosa; CMV showed a second predilection site at the ventricular luminal border . Both microorganisms revealed a strong correlation between each other (r = 0.73; p < 0.001) as well as with hHSP60 (cHSP60: r = 0.74; p < 0.001; CMV: r = 0.80; p < 0.001) . Macrophage infiltration correlated with cHSP60 (r = 0.78; r < 0.001), CMV (r = 0.78; r < 0.001) and hHSP60 (r = 0.56; r = 0.007) . Of note, the frequency of cHSP60, CMV and CD68 signaling was increased more than 5-fold in prosthetic valves compared to native valves (p = 0.017, p = 0.002 and p = 0.005) . In summary, valvular infections of Chlamydia pneumoniae and of cytomegalovirus are frequently seen in degenerated aortic valves, irrespective of native or prosthetic origin . Colocalization of both HSP60 homologues and cytomegalovirus within macrophages in valvular fibrosa points to regional stressor effects that might be at least partly attributable to chronic persistent pathogen burden and molecular mimicry. Biol Psychiatry, 2002 Jun 15, 51(12), 979 - 87 High-avidity human serum antibodies recognizing linear epitopes of Borna disease virus proteins; Billich C et al.; BACKGROUND: The recent observation that Borna disease virus (BDV)-reactive antibodies from psychiatric patients exhibit only low avidity for BDV antigen called into question their diagnostic value and raised the possibility that antigenically related microorganisms or self antigens caused the production of these antibodies . We further characterized the specificity of these antibodies.METHODS: We established a peptide array-based screening test that allows the identification of antibodies directed against linear epitopes of the two major BDV proteins, the nucleoprotein (N) and the phosphoprotein (P).RESULTS: Initial tests employing sera of BDV-infected mice and rats or horses with Borna disease revealed a high specificity and sensitivity of this test . All sera recognized epitopes of N, P, or both . Sera of noninfected rats, mice, and horses showed no signals on either peptide array . Several human sera that recognized BDV antigen by indirect immunofluorescence contained antibodies that recognized various linear epitopes of one or even both BDV proteins . Remarkably, antibodies purified from such human serum by matrix-immobilized peptides showed high-avidity binding to BDV antigens when assayed by IFA or Western blotting.CONCLUSIONS: These data suggest that reactive antibodies found in psychiatric patients indeed indicate infection with BDV or a BDV-like agent . However, the poor affinity maturation of BDV-specific human antibodies remains unexplained. Am J Cardiol, 2002 Jun 15, 89(12), 1400 - 4 Usefulness of cytokines interleukin-6 and interleukin-2R concentrations in diagnosing active infective endocarditis involving native valves; Alter P et al.; The most important diagnostic value in infective endocarditis (IE) is isolation of the causative microorganism . Because premature antibiotic treatment is commonly administered before the assessment of blood cultures, the percentages of isolated microorganisms has decreased significantly within the last decades . Therefore, additional criteria for the diagnosis of IE may be helpful . It was hypothesized that assessment of interleukin-6 (IL-6) and interleukin-2R (IL-2R) may provide new diagnostic criteria for inflammation in IE . IL-6 and IL-2R serum concentrations, white blood cell count (WBC), and C-reactive protein (CRP) were measured in the blood of 47 patients with IE at the time of diagnosis and during treatment . WBC and CRP were elevated in patients with IE at the time of diagnosis . Both parameters were higher (p <0.05) in patients with positive blood cultures when compared with negative cultures . The differences persisted during the first week of treatment (p <0.01) . In contrast, IL-6 and IL-2R concentrations were elevated (p <0.001) independently of the status of blood cultures . Serum concentrations of IL-6 and IL-2R decreased continuously during antibiotic treatment . Assessment of IL-6 and IL-2R could thus provide new diagnostic criteria for inflammation in IE, and these interleukins could also be suitable for monitoring the course of inflammation during treatment. Bratisl Lek Listy . 2002;103(2):94. Retrobulbar neuritis associated with Borrelia afzelii infection; Strmen P et al.; PURPOSE: To report retrobulbar neuritis caused by Borrelia afzelii culturally proved from cerebrospinal fluid (CSF) . METHODS: A 23 year old female underwent ophthalmologic, laboratory and other auxilliary examinations . RESULTS: CSF cultures grew spirochetal microorganisms, serotyped by monoclonal antibodies as Borrelia afzelii . Following the serological and cultural results, treatment with doxycycline 200 mg daily was started and kept for three weeks . Gradual improvement of the visual acuity of the right eye was observed with full recovery to 20/20 . CONCLUSIONS: Borrelia infection should be considered in the differential diagnosis of retrobulbar neuritis . CSF should be examined also culturally . (Ref . 5.) Proc Natl Acad Sci U S A, 2002 Jun 11, 99(12), 7968 - 73 The biosynthetic gene cluster of the maytansinoid antitumor agent ansamitocin from Actinosynnema pretiosum; Yu TW et al.; Maytansinoids are potent antitumor agents found in plants and microorganisms . To elucidate their biosynthesis at the biochemical and genetic level and to set the stage for their structure modification through genetic engineering, we have cloned two gene clusters required for the biosynthesis of the maytansinoid, ansamitocin, from a cosmid library of Actinosynnema pretiosum ssp . auranticum ATCC 31565 . This is a rare case in which the genes involved in the formation of a secondary metabolite are dispersed in separate regions in an Actinomycete . A set of genes, asm22-24, asm43-45, and asm47, was identified for the biosynthesis of the starter unit, 3-amino-5-hydroxybenzoic acid (AHBA) . Remarkably, there are two AHBA synthase gene homologues, which may have different functions in AHBA formation . Four type I polyketide synthase genes, asmA-D, followed by the downloading asm9, together encode eight homologous sets of enzyme activities (modules), each catalyzing a specific round of chain initiation, elongation, or termination steps, which assemble the ansamitocin polyketide backbone . Another set of genes, asm13-17, encodes the formation of an unusual "methoxymalonate" polyketide chain extension unit that, notably, seems to be synthesized on a dedicated acyl carrier protein rather than as a CoA thioester . Additional ORFs are involved in postsynthetic modifications of the initial polyketide synthase product, which include methylations, an epoxidation, an aromatic chlorination, and the introduction of acyl and carbamoyl groups . Tentative functions of several asm genes were confirmed by inactivation and heterologous expression. Proc Natl Acad Sci U S A, 2002 Jun 25, 99(13), 8654 - 9 Epub 2002 Jun 11. Tuning the membrane surface potential for efficient toxin import; Zakharov SD et al.; Membrane surface electrostatic interactions impose structural constraints on imported proteins . An unprecedented sensitive dependence on these constraints was seen in the voltage-gated import and channel formation by the C-terminal pore-forming domain of the bacteriocin, colicin E1 . At physiological ionic strengths, significant channel current was observed only in a narrow interval of anionic lipid content ({L-}), with the maximum current (I(max)) at 25-30 mol% (dioleoyl)-phosphatidylglycerol ({L-}max) corresponding to a surface potential of the lipid bilayer in the absence of protein, psi(o)max = -60 +/- 5 mV . Higher ionic strength shifted {L-}max to larger values, but psi(o)max remained approximately constant . It is proposed that the channel current (i) increases and (ii) decreases at /psi(o)/ values <55 mV and >65 mV, because of (i) electrostatic interactions needed for effective insertion of the channel polypeptide and (ii) constraints due to electrostatic forces on the flexibility needed for cooperative insertion into the membrane . The loss of flexibility for /psi(o)/ 65 mV was demonstrated by the absence of thermally induced intraprotein distance changes of the bound polypeptide . The anionic lipid content, 25-30 mol%, corresponding to the channel current maxima, is similar to that of the target Escherichia coli cytoplasmic membrane and membranes of mesophilic microorganisms . This suggests that one reason the membrane surface potential is tuned in vivo is to facilitate protein import. Physiol Plant, 2001 Nov, 113(3), 301 - 307 An aquatic perspective on the concepts of Ingestad relating plant nutrition to plant growth; Raven JA; The Ingestad approach to the culture of higher terrestrial plants for physiological studies is discussed in relation to a number of resources, organisms and growth situations that were not part of the original design and rationale of Ingestad's methodology . The additional resource considered is photosynthetically active radiation, and difficulties of applying the Ingestad approach to this resource as well as to atmospheric CO2 are considered . The relationship of the Ingestad approach to reductionist studies based on enzyme kinetic studies is then briefly considered . The organisms considered next are aquatic plants, including both micro- and macrophytes . The consideration of photosynthetic microorganisms leads to a comparison of the Ingestad approach with growth in batch, and in continuous (chemostat and turbidostat) cultures, and with studies on growth in synchronous cultures in which cyclic changes in cell composition in the cell growth and division cycle can be identified . The natural environmental conditions for these organisms are a natural extension of the light/dark synchronization of laboratory cultures, and the bloom (batch culture equivalent to new production) and of grazing and parasitism removing biomass and recycling nutrients (chemostat or turbidostat culture equivalent to recycled production) situations for phytoplankton . The overall conclusion is that, while the Ingestad approach is a useful mirror in which to examine other concepts of plant resource acquisition and manipulation, the Ingestad methodology seems to make assumptions about the intrinsic growth rate and composition of plants that cannot be independently verified. Int Endod J, 2002 Apr, 35(4), 366 - 71 Invasion of vascular cells in vitro by Porphyromonas endodontalis; Dorn BR et al.; AIM: The objective of this study was to determine whether laboratory strains and clinical isolates of microorganisms associated with root canal infections can invade primary cultures of cardiovascular cells . METHODOLOGY: Quantitative levels of bacterial invasion of human coronary artery endothelial cells (HCAEC) and coronary artery smooth muscle cells (CASMC) were measured using a standard antibiotic protection assay . Transmission electron microscopy was used to confirm and visualize internalization within the vascular cells . RESULTS: Of the laboratory and clinical strains tested, only P . endodontalis ATCC 35406 was invasive in an antibiotic protection assay using HCAEC and CASMC . Invasion of P . endodontalis ATCC 35406 was confirmed by transmission electron microscopy . DISCUSSION: Certain microorganisms associated with endodontic infections are invasive . If bacterial invasion of the vasculature contributes to the pathogenesis of cardiovascular disease, then microorganisms in the pulp chamber represent potential pathogens. Trans R Soc Trop Med Hyg, 2002 Apr, 96 Suppl 1, S9 - 13 Evolutionary genetics and molecular diagnosis of Leishmania species; Banuls AL et al.; An extensive study has been performed on various natural populations of Leishmania from the 'Old' and 'New Worlds' using multilocus enzyme electrophoresis and random amplification of polymorphic deoxyribonucleic acid . The data are interpreted in evolutionary genetic terms in order to give a firm basis to studies dealing with the relevant medical properties of pathogens . We confirm that Leishmania undergoes clonal evolution with occasional phenomena of hybridization . This suggests that the microorganism genotypes are stable in space and time and consequently have epidemiological and medical relevance . It is crucial to have a clear definition of the taxa to be identified . In the case of Leishmania, there is at present no firm consensus on the species concept . We propose that any new species of Leishmania should correspond to a 'discrete typing unit' that exhibits specific medical and/or epidemiological characters . Based on this approach, the species status of L . peruviana can be supported . On the contrary, L . panamensis cannot be clearly distinguished from L . guyanensis . Our studies on the genetic diversity of Leishmania show that a given stock is not representative of the genus, the subgenera, or any species . We suggest that the genetic polymorphism of Leishmania has a strong impact on the parasite's biomedical properties. Arch Biochem Biophys, 2002 Apr 15, 400(2), 273 - 81 Reduction of iron by extracellular iron reductases: implications for microbial iron acquisition; Cowart RE; The extracellular enzymatic reduction of iron by microorganisms has not been appropriately considered . In this study the reduction and release of iron from ferrioxamine were examined using extracellular microbial iron reductases and compared to iron mobilization by chemical reductants, and to chelation by EDTA and desferrioxamine . A flavin semiquinone was formed during the enzymatic reduction of ferrioxamine, which was consistent with the 1 e(-) reduction of iron by an enzyme . The rates for the enzymatic reactions were substantially faster than both the 2 e(-) chemical reductions and the chelation reactions . The rapid rates of the enzymatic reduction reactions demonstrated that these enzymes are capable of accomplishing the extracellular mobilization of iron required by microorganisms . The data suggest that mechanistically there are two phases for the mobilization and transport of iron by those microorganisms that produce both extracellular iron reductases and siderophores, with reduction being the principle pathway. Int J Syst Evol Microbiol, 2002 May, 52(Pt 3), 719 - 28 Geoglobus ahangari gen . nov., sp . nov., a novel hyperthermophilic archaeon capable of oxidizing organic acids and growing autotrophically on hydrogen with Fe(III) serving as the sole electron acceptor; Kashefi K et al.; A novel, regular to irregular, coccoid-shaped, anaerobic, Fe(III)-reducing microorganism was isolated from the Guaymas Basin hydrothermal system at a depth of 2000 m . Isolation was carried out with a new technique using Fe(III) oxide as the electron acceptor for the recovery of colonies on solid medium . The isolate, designated strain 234T, was strictly anaerobic and exhibited a tumbling motility . The cells had a single flagellum . Strain 234T grew at temperatures between 65 and 90 degrees C, with an optimum at about 88 degrees C . The optimal salt concentration for growth was around 19 g l(-1) . The isolate was capable of growth with H2 as the sole electron donor coupled to the reduction of Fe(III) without the need for an organic carbon source . This is the first example of a dissimilatory Fe(III)-reducing micro-organism capable of growing autotrophically on hydrogen . In addition to molecular hydrogen, strain 234T oxidizes pyruvate, acetate, malate, succinate, peptone, formate, fumarate, yeast extract, glycerol, isoleucine, arginine, serine, glutamine, asparagine, stearate, palmitate, valerate, butyrate and propionate with the reduction of Fe(III) . This isolate is the first example of a hyperthermophile capable of oxidizing long-chain fatty acids anaerobically . Isolate 234T grew exclusively with Fe(III) as the sole electron acceptor . The G+C content was 58.7 mol% . Based on detailed analysis of its 16S rDNA sequence, G+C content, distinguishing physiological features and metabolism, strain 234T is proposed to represent a novel genus within the Archaeoglobales . The name proposed for strain 234T is Geoglobus ahangari gen . nov., sp . nov.. Acta Haematol, 2002, 107(4), 220 - 3 Bacteria in blood smears: overwhelming sepsis or trivial contamination; van der Meer W et al.; It is unusual to find microorganisms in peripheral blood smears, and their presence is frequently associated with overwhelming sepsis and consequently a poor prognosis . In this report, we demonstrate 4 cases with bacteria in blood smears . Two of them had a fatal outcome, but the other 2 were caused by a contamination either via the central venous catheter or in vitro, both without dramatic outcome . The finding of bacteria in blood smears has to be interpreted carefully, and thorough examination of peripheral blood smears may be of great importance in the early diagnosis of bacteremia; however, in vitro contamination must be excluded . J Mol Biol, 2002 Apr 26, 318(2), 273 - 85 Transfer RNA gene-targeted retrotransposition of Dictyostelium TRE5-A into a chromosomal UMP synthase gene trap; Beck P et al.; The genome of the eukaryotic microorganism Dictyostelium discoideum hosts a family of seven non-long terminal repeat retrotransposons (TREs) that show remarkable insertion preferences near tRNA genes . We developed an in vivo assay to detect tRNA gene-targeted retrotransposition of endogenous TREs in a reporter strain of D . discoideum . A tRNA gene positioned within an artificial intron was placed into the D . discoideum UMP synthase gene . This construct was inserted into the D . discoideum genome and presented as a landmark for de novo TRE insertions . We show that the tRNA gene-tagged UMP synthase gene was frequently disrupted by de novo insertions of endogenous TRE5-A copies, thus rendering the resulting mutants resistant to 5-fluoroorotic acid selection . Approximately 96% of all isolated 5-FOA-resistant clones contained TRE5-A insertions, whereas the remaining 4% resulted from transposition-independent mutations . The inserted TRE5-As showed complex structural variations and were found about 50 bp upstream of the reporter tRNA gene, similar to previously analysed genomic copies of TRE5-A . No integration by other members of the TRE family was observed . We found that only 51% of the de novo insertions were derived from autonomous TRE5-A.1 copies . The remaining 49% of new insertions were due to TRE5-A.2 elements, which lack the proteins required for reverse transcription and integration, but retain functional promoter sequences . (c) 2002 Elsevier Science Ltd. Int Microbiol, 2001 Dec, 4(4), 227 - 36 Cyanobacterial diversity in extreme environments in Baja California, Mexico: a polyphasic study; Lopez-Cortes A et al.; Cyanobacterial diversity from two geographical areas of Baja California Sur, Mexico, were studied: Bahia Concepcion, and Ensenada de Aripez . The sites included hypersaline ecosystems, sea bottom, hydrothermal springs, and a shrimp farm . In this report we describe four new morphotypes, two are marine epilithic from Bahia Concepcion, Dermocarpa sp . and Hyella sp . The third, Geitlerinema sp., occurs in thermal springs and in shrimp ponds, and the fourth, Tychonema sp., is from a shrimp pond . The partial sequences of the 16S rRNA genes and the phylogenetic relationship of four cyanobacterial strains (Synechococcus cf . elongatus, Leptolyngbya cf . thermalis, Leptolyngbya sp., and Geitlerinema sp.) are also presented . Polyphasic studies that include the combination of light microscopy, cultures and the comparative analysis of 16S rRNA gene sequences provide the most powerful approach currently available to establish the diversity of these oxygenic photosynthetic microorganisms in culture and in nature. Acta Chir Belg, 2002 Apr, 102(2), 75 - 7 Bacterial translocation as a cause for septic complications in obstructive colonic ileus; Belyansky LS et al.; Failure of intestinal barrier function and subsequent translocation of microorganisms and their degradation products play a decisive role in development of systemic septic complications for many systemic and intra-abdominal pathologies, for example, following obstructive colonic ileus (OCI) . This study was aimed at the evaluation of the intestinal barrier state in OCI . Sixty albino Wistar rats weighting 250 to 300 g (mean 265 g) were divided into four groups (15 animals in each) . Acute colonic ileus (ACI) was modeled as follows except a control group (Group 1) . Our objective was to examine changes in bacterial flora in the abdomen, mesenteric lymphatic nodes (MLN), liver, spleen, and lungs during the model of OCI after 72 hours following the beginning of experiment . The composition of parietal mucus in normal and in OCI 48 hours following the beginning of experiment examined . Interleukin (IL-VI) levels were determined in both portal and peripheral blood . The right-hand half of colon was ligated at the level of ileocaecal junction in animals of Group 2 (n = 15), whereas in animals of Group 3 (n = 15) it was ligated at the level of sigmoid colon . With the same purpose, a portion of the suspended caecal content was administered into lumen of the jejunum at a concentration of 10(6) colony-forming units (CFU) in animals of Group 4 (n = 15) . Experimentally--induced OCI causes significant bacterial translocation (BT) in rats . The process of colonization of the proximal small intestine with colonic flora takes place under the conditions of ileus . The conditions favorable for the development of BT are generated with colonization of 10(6) CFU in volume . As a result, intestinal flora penetrates into the abdominal organs and lungs . Its highest concentrations are noted in the lymph nodes, lungs and liver . The modeling of the small intestine colonization with colonic flora (Group 4) demonstrates critical parameters of microbial semination. IUBMB Life, 2002 Feb, 53(2), 125 - 9 Animals are dependent on preformed alpha-amino nitrogen as an essential nutrient; Katagiri M et al.; It has traditionally been thought that animals can utilize ammonia for amino acid biosynthesis, and that for them some amino acids are nutritionally nonessential . Presumably this idea originates from the notions of Schoenheimer (G . L . Foster et al . {1939} J . Biol . Chem . 127, 319-327) and of Rose (W . C . Rose et al . {1948} J . Biol . Chem . 176, 753-762), which we question for the following reasons . First, Schoenheimer's experiments only showed the incorporation of ammonia into amino acids . This may occur simply as an exchange between ammonia and the alpha-amino group of endogenous amino acids and reflects the enzymatic properties of glutamate dehydrogenase, which is a reversible enzyme . Second, Rose's nutritional experiments were concerned with whether carbon skeletons of particular amino acids can (nonessential) or cannot (essential) be synthesized from common intermediates of carbohydrate metabolism . We propose that mammals, living as they do at the top of the food web, are absolutely dependent directly or indirectly on higher plants and microorganisms for preformed alpha-amino nitrogen per se and that the first joining of C- and N-atoms to make glutamate constitutes a basic anabolic system in nature after the fixation of CO2 and N2. Arch Pathol Lab Med, 1996 Nov, 120(11), 1009 - 14 Extraneous tissue in surgical pathology: a College of American Pathologists Q-Probes study of 275 laboratories; Gephardt GN et al.; OBJECTIVE: To develop a multi-institutional reference database of extraneous tissue (contaminants) in surgical pathology . DESIGN: In 1994, participants in the College of American Pathologists Q-Probes quality improvement program performed prospective and retrospective evaluations of extraneous tissue found in surgical pathology microscopic sections for a period of 4 weeks or until 1000 slides were reviewed in each participating laboratory . PARTICIPANTS: Two hundred seventy-five surgical pathology laboratories institutions, predominantly from North America . MAIN OUTCOME MEASURES: Extraneous tissue contamination rate for slides in prospective and retrospective reviews; staffing and practice procedures; location of extraneous tissue on slides; type of extraneous tissue (normal, abnormal, nonneoplastic, neoplasm, microorganisms, etc); class of extraneous tissue (slide or block contaminants); source of extraneous tissue (different or same case); origin of extraneous tissue (pathology laboratory, physician's office or operating room); and degree of diagnostic difficulty caused by extraneous tissue . RESULTS: Three hundred twenty-one thousand seven hundred fifty-seven slides were reviewed in the prospective study and 57083 slides in the retrospective study . There was an overall extraneous tissue rate of 0.6% of slides (2074/321757) in the prospective study and 2.9% of slides (1653/57083) in the retrospective study . Of those slides with extraneous tissue, the extraneous tissue was located near diagnostic tissue sections in 59.5% of the slides reviewed prospectively and in 25.3% of slides reviewed retrospectively; deeper sections were performed to evaluate extraneous tissue in 12.2% of prospective cases and in 3.1% of retrospective cases . Of the laboratories, 98% had written guidelines for changing solution in tissue processors, and 64.9% had guidelines for maintaining water baths free of extraneous tissue . A total of 98.9% used lens paper, filter bags, or sponges for processing fragmented and small specimens . Written protocols for documentation of extraneous tissue in surgical pathology reports were established in 6.1% of laboratories, for removal of extraneous tissue from blocks in 5.7%, and for removal of extraneous tissue from microscopic slides in 4.7% . In 24% of laboratories no comment or record was kept to document extraneous tissue . Extraneous tissue consisted of neoplasm in 12.7% of the prospectively reviewed slides and in 6.0% of the retrospectively reviewed slides . For the prospective study, 59.4% of extraneous tissue was classified as slide contaminants, and 28.4% was found to be contaminants within the paraffin block; for the retrospective study, 72.9% was classified as slide contaminants and 15.9% as block contaminants . For the prospective study, 63.2% of extraneous tissue was presumed to be from a different case, and in the retrospective study, 48.5% was presumed to be from a different case . Over 90% of extraneous tissue was thought to originate from the pathology laboratory . The degree of diagnostic difficulty caused by extraneous tissue was judged to be severe in 0.4% of slides in the prospective study and 0.1% of slides in the retrospective study . In the prospective study, it could not be determined whether the tissue in the diagnostic sections was extraneous in 0.6% of slides, and in the retrospective study, it could not be determined whether tissue in the diagnostic sections was extraneous in 0.1% . CONCLUSIONS: This study has documented the frequency, type, origin, source, and diagnostic difficulty of extraneous tissue and presents benchmarks of extraneous tissue experienced in the general practice of surgical pathology. Cad Saude Publica, 2002 May-Jun, 18(3), 873 - 5 {What is the source of mycelial fungi in expressed human milk?}; Novak FR et al.; The authors characterized the genera of mycelial fungi found in samples of expressed human milk received through home collection by the Human Milk Bank of the Instituto Fernandes Figueira in Rio de Janeiro . A total of 821 samples of expressed human milk were taken randomly from bottles collected at home by the milk donors themselves and were investigated for molds, yeasts, and mesophilic microorganisms . The analyses showed the occurrence of molds and yeasts in 43 (5.2%) of the samples, with counts reaching 103CFU/ml . Some 48 strains of mycelial fungi were identified by standard laboratory techniques, including: Aspergillus Niger group (6.3%), Aspergillus sp . (4.2%), Paecilomyces sp . (12.6%), Penicillium sp . (60.4%), Rhizopus sp . (2.0%), and Syncephalastrum sp . (14.5%) . The authors discuss the importance of donor hands' asepsis prior to collecting human milk. Pediatr Dermatol, 2002 May-Jun, 19(3), 232 - 6 Fatal septicemia in an infant with keratitis, ichthyosis, and deafness (KID) syndrome; Gilliam A et al.; Keratitis, ichthyosis, and deafness (KID) syndrome is a rare congenital disorder of unknown etiology in which increased susceptibility to viral, bacterial, and mycotic infections has been observed . We report an infant with KID syndrome who died from overwhelming systemic infection . To date, investigations into the immune function of patients with this syndrome have not revealed a common underlying systemic immune deficit . However, the severity of infections and multiplicity of organisms observed in this syndrome suggest that a primary immunodeficiency is present in addition to an impaired cutaneous barrier to microorganisms. Naturwissenschaften, 2002 Feb, 89(2), 79 - 83 Midgut tissue of male pine engraver, Ips pini, synthesizes monoterpenoid pheromone component ipsdienol de novo; Hall GM et al.; For over three decades the site and pathways of bark beetle aggregation pheromone production have remained elusive . Studies on pheromone production in Ips spp . bark beetles have recently shown de novo biosynthesis of pheromone components via the mevalonate pathway . The gene encoding a key regulated enzyme in this pathway, 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-R), showed high transcript levels in the anterior midgut of male pine engravers, Ips pini (Say) (Coleoptera:Scolytidae) . HMG-R expression in the midgut was sex, juvenile hormone, and feeding dependent, providing strong evidence that this is the site of acyclic monoterpenoid (ipsdienol) pheromone production in male beetles . Additionally, isolated midgut tissue from fed or juvenile hormone III (JH III)-treated males converted radiolabeled acetate to ipsdienol, as assayed by radio-HPLC . These data support the de novo production of this frass-associated aggregation pheromone component by the mevalonate pathway . The induction of a metazoan HMG-R in this process does not support the postulated role of microorganisms in ipsdienol production. J Environ Sci (China), 2002 Apr, 14(2), 264 - 8 Composting MSW and sewage sludge with effective complex microorganisms; Xi BD et al.; The effects of complex microorganisms in composting process of the municipal solid waste (MSW) and sludge were examined through inspecting biomass, temperature, oxygen consumption, organic mater, and C/N (the ratio of carbon and nitrogen) . The experimental results shows: complex microorganisms are effective to compose organic matter and speedup composting change into humus. Cas Lek Cesk, 2002 Mar 1, 141(4), 107 - 11 {Genetically modified organisms--problems and legislation}; Drobnik J; Genetically modified organisms are defined by law as entities capable of replication and/or transmission of hereditary material that had been altered by the insertion or removal of a DNA fragment . By the EU legal regulation as well as by the Czech law, such organisms are considered risky whereas other products of breeding, though obtained by, e.g., induced mutagenesis, are claimed as safe . Organisms transferred from other ecosystems are also considered safe . The Czech law on the use of genetically modified organisms is based on registers of users and organisms for specific use . Application for the registration that is valid as an approval should be submitted to the Ministry of Environment . The applicant is obliged to present the risk assessment of the particular use of genetically modified organisms . Genetically modified organisms are connected with certain risk to ecology, however health risks are brought about almost exclusively by microorganisms . Modified organisms used for food production are thoroughly tested for substantial equivalency with standard crops and with respect to health parameters of the protein(s) newly introduced due to genetic modification . Detail tests as well as their cost are close to the testing of new drugs . European as well as Czech rules for food labelling are motivated by the psychology of consumers rather than by health impact . They result to absurdities but do not meet the task of public psychology . This is why the EU authorities are looking for measures to change the present situation that other wise would bring Europe well behind the developed countries. Front Biosci, 2002 Jun 01, 7, d1487 - 93 Association of alcohol consumption and exaggerated immunopathologic effects in the liver induced by infectious organism; Jerrells TR; The cause of alcoholic liver disease (ALD) is multifactorial and poorly understood . It is clear that alcohol alone is not responsible for most of the changes associated with ALD and that cofactors are involved in initiation and production of ALD . One cofactor that has received a great deal of attention recently is the concomitant infection with hepatitis C virus (HCV) and alcohol abuse . The interactive effects of HCV and alcohol abuse are still unclear, but apparently they are the result of an inability of the immune system to control the viral infection and exaggerated hepatocyte damage mediated by either the cells of the inflammatory response or factors produced by the inflammatory cells . A major effort in my laboratory has been focused on defining the effects of alcohol consumption on immunity to various infectious agents . Efforts have also been directed to elucidating the pathologic effects in the liver of inflammatory and immune responses to microorganisms that either specifically or ultimately infect the liver from an initial site of infection other than the liver . This review will focus on one aspect of the possible pathogenic effects associated with alcohol abuse and hepatic infections: the possible role of the immune system, notably the cytotoxic T lymphocyte (CTL) response . It is clear that the development of a CTL response is critical for the control of HCV and other infections, and it is also likely that this response is involved in liver damage . In this review, the evidence that shows the importance of the CD8+ CTL in bacterial and viral clearance and the role for pathogenesis will be presented . Findings obtained from animal studies that support the suggestion that activated CD8+ CTLs can induce liver damage will be presented, as will results of recent studies from my laboratory that provide evidence for an effect of alcohol to enhance the liver damage mediated by activated CD8+ T cells. FEMS Microbiol Lett, 2002 May 7, 210(2), 215 - 9 The effects of volatile microbial secondary metabolites on protein synthesis in Serpula lacrymans; Humphris SN et al.; The effects of volatile secondary metabolites produced by Trichoderma pseudokoningii, Trichoderma viride and Trichoderma aureoviride on growth rate and protein synthesis in two Serpula lacrymans isolates were investigated . Mycelial growth was affected to differing degrees, depending on the specific interactive microbial couplet involved . Protein synthesis by both S . lacrymans (Forfar) and S . lacrymans (H28) was affected by the volatile secondary metabolites of T . aureoviride and T . viride, but not by those of T . pseudokoningii . Mycelial growth and the original pattern of protein synthesis resumed when the antagonists were removed . It is probable that volatile secondary metabolites have played an important role during the evolution of microorganisms in the context of community, population and functional dynamics. Zh Mikrobiol Epidemiol Immunobiol, 2002 Mar-Apr, (2), 9 - 13 {Viability and virulence of Francisella tularensis subsp . Holarctica in water ecosystems (experimental study)}; Mironchuk IuV et al.; Under conditions of artificial water biocenosis a virulent strain of F . tularensis could be detected in fresh water shrimps and mollusks for about a month, in Conepoda for up to 20 days and in Chydorus sphaericus for up to 7 days from the moment of the aquaria water contamination . In silt F . tularensis could be detected for a longer period (up to 2 months) . Daphnia, Oligochaeta and C . sphaericus appeared to be unfavorable environment for this microorganism . The virulence level of F . tularensis microbial cells decreased in paralell with prolongation of their stay in water biocenosis . The presence of water biota favours F . tularensis preservation in water reservoirs for a longer time. Nat Biotechnol, 2002 Jun, 20(6), 567 - 74 Potential for the environmental impact of transgenic crops; Dale PJ et al.; In recent years, there has been increasing interest in how changes in agricultural practice associated with the introduction of particular genetically modified (GM) crops might indirectly impact the environment . There is also interest in any effects that might be associated with recombinant and novel combinations of DNA passing into the environment, and the possibility that they may be taken up by microorganisms or other live biological material . From the current state of knowledge, the impact of free DNA of transgenic origin is likely to be negligible compared with the large amount of total free DNA . We can find no compelling scientific arguments to demonstrate that GM crops are innately different from non-GM crops . The kinds of potential impacts of GM crops fall into classes familiar from the cultivation of non-GM crops (e.g., invasiveness, weediness, toxicity, or biodiversity) . It is likely, however, that the novelty of some of the products of GM crop improvement will present new challenges and perhaps opportunities to manage particular crops in creative ways. J Chemother, 1991 Jan, 3 Suppl 1, 218 - 21 Imipenem use in serious childhood infections; Pavesio D et al.; Imipenem plus cilastatin is a beta-lactam antibiotic with a broad spectrum and good tolerance . For this its use is indicated in serious infections even in children . The authors carried out a preliminary bacteriological study in patients affected by serious infection, cystic fibrosis, tumors, neutropenia, subjects undergoing intensive therapy and those with abdominal surgical infection . Isolated microorganism sensitivity in these patients, towards imipenem was very high (89%-100%) . With these premises, 35 patients with serious infections in various locations were treated only with imipenem . Of these, 15 were oncologic neutropenic and 20 normal . In the first group therapeutic success was 66.6% and in the second one 80% . Local and systemic tolerance always showed good results. Southeast Asian J Trop Med Public Health, 2001, 32 Suppl 2, 183 - 9 Inactivation of Cryptosporidium parvum oocysts in field soil; Kato S et al.; Cryptosporidium parvum oocysts from dairy calves are believed to regularly contaminate watersheds . Identifying oocysts and measuring their viability in the natural environment are important elements in estimating the risk posed by this resistant organism . A 152 day field study was conducted to measure the viabilities of oocysts inoculated into 25 sampling points . Water potential, pH, and ammonium content were also measured at the same 25 sampling sites . A three-dimensional mapping program (Surfer) was used to create 3-D maps of the viabilities of C . parvum oocysts and other factors measured during the experiment . The results indicate that 3-D graphical presentation may be a useful means to identify potential sites of greatest risk of oocyst survival and could indicate areas where natural conditions are causing the most rapid oocyst inactivation, and this method can be a means for the future measurement of microorganism inactivation in the natural environment. Microbiol Mol Biol Rev, 2002 Jun, 66(2), 223 - 49 Genetics and assembly line enzymology of siderophore biosynthesis in bacteria; Crosa JH et al.; The regulatory logic of siderophore biosynthetic genes in bacteria involves the universal repressor Fur, which acts together with iron as a negative regulator . However in other bacteria, in addition to the Fur-mediated mechanism of regulation, there is a concurrent positive regulation of iron transport and siderophore biosynthetic genes that occurs under conditions of iron deprivation . Despite these regulatory differences the mechanisms of siderophore biosynthesis follow the same fundamental enzymatic logic, which involves a series of elongating acyl-S-enzyme intermediates on multimodular protein assembly lines: nonribosomal peptide synthetases (NRPS) . A substantial variety of siderophore structures are produced from similar NRPS assembly lines, and variation can come in the choice of the phenolic acid selected as the N-cap, the tailoring of amino acid residues during chain elongation, the mode of chain termination, and the nature of the capturing nucleophile of the siderophore acyl chain being released . Of course the specific parts that get assembled in a given bacterium may reflect a combination of the inventory of biosynthetic and tailoring gene clusters available . This modular assembly logic can account for all known siderophores . The ability to mix and match domains within modules and to swap modules themselves is likely to be an ongoing process in combinatorial biosynthesis . NRPS evolution will try out new combinations of chain initiation, elongation and tailoring, and termination steps, possibly by genetic exchange with other microorganisms and/or within the same bacterium, to create new variants of iron-chelating siderophores that can fit a particular niche for the producer bacterium. J Antimicrob Chemother, 2002 Jun, 49(6), 897 - 903 Patient compliance with antibiotic treatment for respiratory tract infections; Kardas P; Despite doctors' expectations, non-compliance is common in short-term antibiotic therapy of respiratory tract infections (RTIs) . This phenomenon has profound practical implications . It leads to ineffective management, the deterioration of patients' health, hospital admissions, additional costs and the emergence of antibiotic-resistant microorganisms . This article reviews methods of measuring compliance with antibiotics in the outpatient-based management of RTIs and research results . Causes of non-compliance are also discussed . Factors influencing compliance are analysed, as well as practical strategies for addressing non-compliance with antibiotic therapies for RTIs . The influence of the frequency of doses on compliance is particularly stressed, as it has been observed that once daily dosing has almost a 100% compliance rate . As a number of once-daily antibiotic preparations are available now, the possibility of using once-daily schedules for improving compliance in RTI cases is stressed. Appl Environ Microbiol, 2002 Jun, 68(6), 3108 - 13 Universal immunoprobe for (per)chlorate-reducing bacteria; O'Connor SM et al.; Recent studies in our lab have demonstrated the ubiquity and diversity of microorganisms which couple growth to the reduction of chlorate or perchlorate {(per)chlorate} under anaerobic conditions . We identified two taxonomic groups, the Dechloromonas and the Dechlorosoma groups, which represent the dominant (per)chlorate-reducing bacteria (ClRB) in the environment . As part of these studies we demonstrated that chlorite dismutation is a central step in the reductive pathway of (per)chlorate that is common to all ClRB and which is mediated by the enzyme chlorite dismutase (CD) . Initial studies on CD suggested that this enzyme is highly conserved among the ClRB, regardless of their phylogenetic affiliation . As such, this enzyme makes an ideal target for a probe specific for these organisms . Polyclonal antibodies were commercially raised against the purified CD from the ClRB Dechloromonas agitata strain CKB . The obtained antiserum was deproteinated by ammonium sulfate precipitation, and the antigen binding activity was assessed using dot blot analysis of a serial dilution of the antiserum . The titers obtained with purified CD indicated that the antiserum had a high affinity for the CD enzyme, and activity was observed in dilutions as low as 10(-6) of the original antiserum . The antiserum was active against both cell lysates and whole cells of D . agitata, but only if the cells were grown anaerobically with (per)chlorate . No response was obtained with aerobically grown cultures . In addition to D . agitata, dot blot analysis employed with both whole-cell suspensions and cell lysates of several diverse ClRB representing the alpha, beta, and gamma subclasses of Proteobacteria tested positive regardless of phylogenetic affiliation . Interestingly, the dot blot response obtained for each of the ClRB cell lysates was different, suggesting that there may be some differences in the antigenic sites of the CD protein produced in these organisms . In general, no reactions were observed with cells or cell lysates of the organisms closely related to the ClRB which could not grow by (per)chlorate reduction . These studies have resulted in the development of a highly specific and sensitive immunoprobe based on the commonality of the CD enzyme in ClRB which can be used to assess dissimilatory (per)chlorate-reducing populations in environmental samples regardless of their phylogenetic affiliations. Int J Food Microbiol, 2002 Jun 5, 76(1-2), 127 - 34 Intra-species chromosome-length polymorphism in Geotrichum candidum revealed by pulsed field gel electrophoresis; Gente S et al.; Geotrichum candidum is an ascomycetous anamorph yeast-like fungus found in various habitats . It is a component of the natural flora of milk and is used as a maturing agent for both soft and hard cheeses . This microorganism displays phenotypic variability and may act as an opportunist pathogen, causing geotrichosis . Cytological analysis of G . candidum strain ATCC 204307 showed this strain to have eight chromosomes . We prepared chromosomal DNA from 13 strains of G . candidum differing in habitat and morphotype . We used pulsed field gel electrophoresis (PFGE) in two sets of conditions to determine the size of the chromosomal DNA molecules . The strains investigated had five to eight chromosomes, 0.6 to 4.5 Mb in size . We estimated genome size in these 13 strains to be between 11 and 19 Mb . Pulsed-field gel electrophoresis profiles showed a high degree of polymorphism, indicating considerable variability between strains . Genome size and the presence of large chromosomes appeared to be correlated with morphotype . Strains with a mold-like or intermediate morphotype tended to have larger genomes than strains with a yeast-like morphotype did. Med Pregl, 2002 Jan-Feb, 55(1-2), 19 - 22 {Aminoglycoside antibiotics and post-antibiotic effects}; Stefan-Mikic S et al.; INTRODUCTION: Man has been fighting diseases for centuries . One of the major battles is against microorganisms and diseases they cause . A health education course was organized on prescribing aminoglycoside antibiotics and postantibiotic effect . The aim of the course was to change the prescription habits in our colleagues . The postantibiotic effect of aminoglycoside antibiotics as well as impact of subinhibiting doses on duration of postantibiotic effect requires modification of previous therapeutic protocols . Single daily dose has the same or even greater effect than multiple daily doses . The toxicity of aminoglycosides is not increased and remains the same or smaller in single daily regimens . RESULTS: The single daily dose regimen of aminoglycosides has been used in 63.6% of cases in Clinic for Infectious Diseases of the Clinical Center of Novi Sad, 41.2% in Outpatient Health Care Center of Novi Sad "Liman" and this regimen has not been used in General Practice Department, Children's Health Care Department and Ear, Nose and Throat Clinic at all . The twice daily regimen has been used instead . CONCLUSION: Doctors are aware of the postantibiotic effect, but vast majority are still bound to their old habits in regard to prescribing antibiotics . Our educational course failed to achieve its goal. Intern Med J, 2002 May-Jun, 32(5-6), 242 - 51 Surface-active phospholipid: a Pandora's box of clinical applications . Part II . Barrier and lubricating properties; Hills BA; In Part I, it was described how their configuration renders phospholipid molecules surface active and capable of acting at interfaces in addition to the liquid-air interface to which conventional theory has hitherto confined the study of 'surfactant' in the lung . Surface-active phospholipid (SAPL) appears no different to comparable surfactants studied in the physical sciences for the highly desirable properties that their adsorption (reversible binding) can impart to solid surfaces . In Part II, these properties are considered in sites where there is no air . Highly desirable properties include boundary lubrication (lubricity), release (antistick) and the ability of the strongly adsorbed and strongly cohesive SAPL linings to act as barriers against abrasion, corrosion and, possibly, against invasion by microorganisms . As the 'sealant', it could be the true barrier rather than the cells providing its mechanical support . Evidence is reviewed for SAPL providing the gastric mucosal barrier to acid in the stomach and preventing the digestion of Helicobacter pylori until that barrier is broken by bile in the duodenum, where H . pylori cause ulcers . The concept that SAPL provides effortless sliding of many tissues, including pleura, pericardium and peritoneum is reviewed . Particular attention is paid to the load-bearing joints, where a deficiency has been associated with osteoarthritis . The ability of the same SAPL lining to perform multiple roles is discussed in relation to the peritoneum, where it could provide the lubricant/release agent preventing surgical adhesions, while imparting semipermeability to 'the membrane' vital for peritoneal dialysis . In each site, the prophylactic use of exogenous SAPL is discussed for its potential clinical applications. Biosci Biotechnol Biochem, 2002 Apr, 66(4), 883 - 6 Formation of new lipoaminopeptides, acremostatins A, B, and C, by co-cultivation of Acremonium sp . Tbp-5 and Mycogone rosea DSM 12973; Degenkolb T et al.; Formation of new lipoaminopeptides, acremostatins A, B, and C, was observed during co-cultivation of Acremonium sp . Tbp-5 and Mycogone rosea DSM 12973 . Thus, co-cultivation of microorganisms producing related products could be suggested as a suitable way towards diversification of microbial structures. Biosci Biotechnol Biochem, 2002 Apr, 66(4), 801 - 7 Isolation and characterization of a beta-primeverosidase-like endo-manner beta-glycosidase from Aspergillus fumigatus AP-20; Yamamoto S et al.; A novel beta-glycosidase-producing microorganism was isolated from soil and identified as Aspergillus fumigatus AP-20 based on its taxonomical characteristics . The enzyme was found to be an extracellular protein in the culture of the isolated fungus and was purified 88-fold by fractionation with ammonium sulfate followed by successive column chromatographies on phenyl-Sepharose HP and Mono P HR . The molecular mass was estimated to be 47 kDa by SDS-PAGE and the isoelectric point to be pH 6.0 by isoelectric focusing . The purified enzyme was highly specific for a substrate, p-nitrophenyl beta-primeveroside (6-O-beta-D-xylopyranosyl-beta-D-glucopyranoside), which was cleaved in an endo-manner into primeverose and p-nitrophenol. Life Sci Space Res, 1965, 3, 64 - 73 Response of microorganisms to a simulated Martian environment; Hawrylewicz EJ et al.; A study was undertaken to determine whether terrestrial microorganisms can survive in a simulated Martian environment . The ultimate objective is to establish whether earth organisms can contaminate Mars . In addition, any demonstration of survival and growth in a simulated Martian environment will provide information relating to the biology of Mars . In the experimental design, exhaustive consideration was given to the duplication of the known and the theoretical environmental parameters of Mars . These included composition of the soil and the atmosphere, barometric pressure, moisture content, solar radiation, and diurnal temperature extremes . Based upon these considerations, a simulated Martian summer environment was defined and used in the experiments . One group of microorganisms was selected from culture collections on the basis of their known characteristics . The other group was made of microorganisms isolated from soils . The soil samples were obtained from the Antarctic, from New Mexico, and California deserts, and from the Colorado tundra . The studies showed that a number of microorganisms can survive the simulated Martian environment . However, no substantial growth under such conditions could be demonstrated . The ability of microorganisms to form spores as a mechanism for survival will be discussed . Also, experiments utilizing augmented environments to establish minimum environmental conditions which will permit growth will be described. Am J Respir Cell Mol Biol, 2002 Jun, 26(6), 739 - 47 Porcine surfactant protein D is N-glycosylated in its carbohydrate recognition domain and is assembled into differently charged oligomers; van Eijk M et al.; Surfactant protein D (SP-D) belongs to a subgroup of mammalian collagenous Ca(2+)-dependent lectins known as the collectins . It is thought to play a significant role in the innate immune response against microorganisms within the lungs and at other mucosal surfaces . This report documents the isolation and characterization of SP-D purified from porcine lung lavage using mannan affinity chromatography and gel filtration . Ultrastructural analysis shows both dodecameric and higher order oligomeric complexes of SP-D . The molecular mass of monomeric porcine SP-D (50 kD) is larger than that of SP-D from humans (43 kD) . The difference in mass is due to the presence of an Asparagine-linked glycosylation in the carbohydrate recognition domain of porcine SP-D, which is absent in SP-D of other species investigated so far . Analysis of this carbohydrate moiety indicates that it is a highly heterogeneous, complex type oligosaccharide which is sialylated . The heterogeneity of oligosaccharide sialylation results in the existence of many differently charged porcine SP-D isoforms . The removal of the carbohydrate moiety reduces the inhibitory effect of porcine SP-D on influenza A virus haemagglutination . Therefore, the carbohydrate moiety may influence interactions with pathogens. Microb Ecol, 2001 Feb, 41(2), 149 - 161 Design and Performance of rRNA Targeted Oligonucleotide Probes for in Situ Detection and Phylogenetic Identification of Microorganisms Inhabiting Acid Mine Drainage Environments; Bond PL et al.; At Iron Mountain, CA, there is an extreme occurrence of acid mine drainage (AMD) . This is a result of past mining activity that has exposed a sulfide ore body to weathering and microbial activity . This study presents seven new oligonucleotide probes for the detection of microorganisms at this AMD site by fluorescent in situ hybridization . In the design of these probes we have accounted for a large body of 16S rRNA sequence data recently compiled by us . This was obtained by PCR and cloning directly from environmental DNA and was mostly represented by novel sequences . The probes were developed to include detection of novel and uncultivated organisms . This includes detection for the Thermoplasmales group, a new group of Leptospirillum, the genus Sulfobacillus, the Acidiphilium genus, Acidimicrobium and relatives, and for organisms within the delta Proteobacteria . These probes have been used to examine the abundance and distribution of organisms, including novel and uncultivated taxa, and to clarify their potential contributions to AMD production at the site . We anticipate that these probes will be useful tools for exploration of the microbiology of other natural acidic environments and bioleaching systems. Microb Ecol, 2001 Feb, 41(2), 114 - 123 Distribution of Nitrogen-Fixing Microorganisms along the Neuse River Estuary, North Carolina; Affourtit J et al.; Nitrogen fixation genes (nifH) were amplified and sequenced from DNA extracted from surface water samples collected from six stations along the length of the Neuse River Estuary, North Carolina, in order to determine the distribution of nitrogen-fixing organisms in the transition from fresh- to saltwater . Nitrogenase genes were detected in all samples by a nested polymerase chain reaction method, and the amplification products from the upriver, midriver, and downriver stations were cloned, sequenced, and used for phylogenetic analysis . The composition of nifH clone libraries from upriver, midriver, and downriver stations (each composed of 14 randomly selected clones) were very diverse (samples from upriver and midriver stations were composed of 14 unique sequences, downriver station composed of 7 unique sequences) and differed among the stations . Some phylotypes were found at more than one station, but were usually found in the upriver and midriver stations or in the midriver and downriver stations, indicating that the phylotypes were probably transported along the river . Cyanobacterial nifH were not found at the most upriver site, but were a large fraction of sequences (50%) recovered from the downriver station, where nitrate concentration was an order of magnitude lower and salinity was higher . In contrast, g proteobacteria nifH sequences were much more common at the midriver and upriver sites (58% and 64%, respectively), compared to the downriver site (14%) . Results indicate that substantially different nitrogen-fixing assemblages are present along the river, reflecting differential watershed hydrological inputs, sedimentation, and environmental selection pressures, along the salinity gradient. Arch Orthop Trauma Surg, 2002 May, 122(4), 204 - 11 Epub 2002 Apr 06. Arthroscopic treatment of septic joints: prognostic factors; Vispo Seara JL et al.; From 1987 to 1993, 88 patients (average age 48 years, range 3 months to 83 years) with septic arthritis were treated at the orthopaedic clinic Konig Ludwig Haus of the University of Wurzburg by arthroscopic means consisting of joint debridement and application of suction drains, combined with appropriate antibiotics and early functional treatment . The series consisted of 78 knees, 8 shoulders and 3 ankles . At the first visit to our clinic, patients typically presented with fever, leucocytosis, elevated sedimentation rate and localized findings in almost every joint involved (generalized tenderness, swelling, effusion, painful and limited range of motion) . All the patients were taken to the operating room on an emergency basis . Broad-spectrum antibiotics were given before the initial culture was evaluated, and its sensitivity was determined . Recovery from arthritis by elimination of joint effusion and disappearance of the inflammatory syndrome occurred in all the joints except for 3 (1 knee joint and 2 shoulders) . The number of arthroscopic procedures needed to become free from infection depended on the one hand on the time between onset of symptoms and arthroscopic surgery and on the other on the kind of microorganism discovered . At the average follow-up evaluation after 2.5 years (range 6 months to 5 years), the functional results were excellent or good in 61% of the patients, satisfactory in 20% and poor in 19% from our series . This functional outcome depends on the degenerative changes of the joint before infection, patient's age, and the time interval between onset of symptoms and surgical intervention; indirectly, the time lapse between the onset of infection and surgical intervention also had an influence on the intraoperative macroscopic appearance. J Periodontol, 2002 May, 73(5), 494 - 500 Association between periodontal pockets and elevated cholesterol and low density lipoprotein cholesterol levels; Katz J et al.; BACKGROUND: Periodontitis is associated with increased prevalence of cardiovascular morbidity and mortality; however, the nature of this association is unclear . There is a rationale that indicates that the presence of periodontal pockets which can harbor pathogenic microorganisms and evoke a host response could elicit a systemic effect . The hypothesis of this study is that periodontal pockets may be associated with elevated blood lipid levels, a known risk factor for atherosclerotic disease . METHODS: The periodontal health of 10,590 Israeli military service men and women was assessed using the Community Periodontal Index of Treatment Needs (CPITN) . The relationship of blood lipids and periodontal disease and CPITN index was tested, controlling for factors that are related to elevated cholesterol levels, including high body mass index (BMI), age, diastolic blood pressure, and smoking . RESULTS: The presence of periodontal pockets was positively associated with higher cholesterol and low density lipoprotein (LDL) cholesterol blood levels in men . No significant association was found in women . CONCLUSIONS: In this large cohort study, the presence of periodontal pockets as measured by CPITN was positively associated with total cholesterol and LDL-cholesterol . The findings of the study support the reports linking increased prevalence of cardiovascular mortality among patients with periodontal disease. J Environ Qual, 2002 May-Jun, 31(3), 997 - 1003 A simple numerical model for predicting organic matter decomposition in a fed-batch composting operation; Nakasaki K et al.; Using dog food as a model of the organic waste that comprises composting raw material, the degradation pattern of organic materials was examined by continuously measuring the quantity of CO2 evolved during the composting process in both batch and fed-batch operations . A simple numerical model was made on the basis of three suppositions for describing the organic matter decomposition in the batch operation . First, a certain quantity of carbon in the dog food was assumed to be recalcitrant to degradation in the composting reactor within the retention time allowed . Second, it was assumed that the decomposition rate of carbon is proportional to the quantity of easily degradable carbon, that is, the carbon recalcitrant to degradation was subtracted from the total carbon remaining in the dog food . Third, a certain lag time is assumed to occur before the start of active decomposition of organic matter in the dog food; this lag corresponds to the time required for microorganisms to proliferate and become active . It was then ascertained that the decomposition pattern for the organic matter in the dog food during the fed-batch operation could be predicted by the numerical model with the parameters obtained from the batch operation . This numerical model was modified so that the change in dry weight of composting materials could be obtained . The modified model was found suitable for describing the organic matter decomposition pattern in an actual fed-batch composting operation of the garbage obtained from a restaurant, approximately 10 kg d(-1) loading for 60 d. Hindustan Antibiot Bull, 1999 Feb-Nov, 41(1-4), 1 - 14 D-amino acid oxidase: its potential in the production of 7-aminocephalosporanic acid; Mujawar SK; D-Amino acid oxidase (DAAO) used in the preparation of alpha-keto acids, in the determination of D-amino acids and in the resolution of racemic mixture of amino acids is produced by a wide range of microorganisms . In the recent past this enzyme is being recognized for its potential in the commercial production of 7-aminocephalosporanic acid (7-ACA), a starting material for various semisynthetic cephalosporins . Though this enzyme is widespread among microorganisms, very few microbial species have been explored for the production of 7-ACA; this is because cephalosporin C is quantitatively deaminated by limited microbial DAAOs . Comparison of physico-chemical properties of enzyme preparations indicate wide variations, however in general DAAOs are specific for D-configuration of amino acids . Both immobilized enzyme and cell preparations are developed for its various applications . The advantages of DAAO in the production of 7-ACA are discussed. Nature, 2002 May 23, 417(6887), 432 - 6 Recovery of 16S ribosomal RNA gene fragments from ancient halite; Fish SA et al.; During the last decade, sensitive techniques for detecting DNA have been successfully applied to archaeological and other samples that were a few hundred to a few thousand years old . Nevertheless, there is still controversy and doubt over claims of exceptionally ancient DNA . Additional accounts stretching back nearly a century suggest that microorganisms may survive over geological time in evaporite deposits . There is, however, often doubt over the age relationship between evaporite formation and the incorporation of microorganisms . Here, we have used petrographic and geochemical techniques (laser ablation microprobe inductively coupled plasma mass spectrometry) to verify the estimated geological age of halite (NaCl) evaporite samples . Fragments of 16S ribosomal RNA genes were detected by polymerase chain reaction amplification of DNA extracted from halite samples ranging in age from 11 to 425 Myr (millions of years) . Haloarchaeal 16S rDNA amplicons were present in one sample (11 16 Myr), whereas other samples (65 425 Myr) yielded only bacterial 16S rDNA amplicons . Terminal restriction fragment length polymorphism analyses indicate complex and different populations of microorganisms or their free DNA in ancient halites of different ages. J Infect Dis, 2002 Jun 1, 185(11), 1637 - 43 Epub 2002 May 17. The molecular basis of sulfonamide resistance in Toxoplasma gondii and implications for the clinical management of toxoplasmosis; Aspinall TV et al.; Polymerase chain reaction amplification and DNA sequencing of the Toxoplasma gondii dihydropteroate synthase gene (dhps) identified 4 alleles among parasite populations from 32 cases of human toxoplasmosis . Heterologous expression and enzyme assay reveal that 3 of these alleles encode sulfadiazine (Sdz)-sensitive enzymes . The fourth, generating a highly Sdz-resistant enzyme, differs from 1 of the other 3 at only a single residue (407) of DHPS . Of interest, a fifth allele, found in a laboratory-induced Sdz-resistant line, also differs from another of these 3 drug-sensitive forms by the same single mutation that affects residue 407 of DHPS . Significantly, residues corresponding to DHPS-407 are implicated in sulfonamide resistance in other microorganisms . The human-derived allelic form encoding the Sdz-resistant enzyme was found in T . gondii associated with a fatal infection, and its presence within clinical material may have implications for sulfonamide use, particularly in cases of toxoplasmosis in which the initial response to drug treatment is poor. J Infect Dis, 2002 Jun 1, 185(11), 1614 - 20 Epub 2002 May 06. Correlation of local interleukin-8 with immunoglobulin A against Gardnerella vaginalis hemolysin and with prolidase and sialidase levels in women with bacterial vaginosis; Cauci S et al.; Mucosal immune system activation may represent a critical determinant of adverse consequences associated with bacterial vaginosis (BV), such as sexual human immunodeficiency virus transmission, upper genital tract infections, postsurgical infections, and adverse pregnancy outcomes . Concentrations of sialidase, prolidase, and anti-Gardnerella vaginalis hemolysin (Gvh) immunoglobulin A (IgA) were higher in vaginal fluids of 75 fertile women with BV, compared with concentrations in vaginal fluids of 85 healthy control subjects . Interleukin (IL)-8 levels were positively associated with anti-Gvh IgA response and inversely correlated with high levels of prolidase and sialidase in women with BV . IL-8 concentration was strongly associated with leukocyte count in both healthy and BV-positive women . The absence of leukocytes in most women with BV likely is due to lack of IL-8 induction . Parallel impairment of innate and adaptive mucosal immune factors, likely through microbial hydrolytic effects, may allow for the ascent of microorganisms to the upper genital tract and may facilitate viral infections. BMC Evol Biol . 2002 May 03;2(1):9. The diversity of microorganisms associated with Acromyrmex leafcutter ants; Van Borm S et al.; BACKGROUND: Molecular biological techniques are dramatically changing our view of microbial diversity in almost any environment that has so far been investigated . This study presents a systematic survey of the microbial diversity associated with a population of Acromyrmex leafcutter ants . In contrast to previous studies on social insects, which targeted specific groups of symbionts occurring in the gut (termites, Tetraponera ants) or in specialised cells (Camponotus ants) the objective of our present study was to do a total screening of all possible micro-organisms that can be found inside the bodies of these leafcutter ants . RESULTS: We amplified, cloned and sequenced SSU rRNA encoding gene fragments from 9 microbial groups known to have insect-associated representatives, and show that: (1) representatives of 5 out of 9 tested groups are present, (2) mostly several strains per group are present, adding up to a total of 33 different taxa . We present the microbial taxa associated with Acromymex ants in a phylogenetic context (using sequences from GenBank) to assess and illustrate to which known microorganisms they are closely related . The observed microbial diversity is discussed in the light of present knowledge on the evolutionary history of Acromyrmex leafcutter ants and their known mutualistic and parasitic symbionts . CONCLUSIONS: The major merits of the screening approach documented here is its high sensitivity and specificity, which allowed us to identify several microorganisms that are promising candidates for further study of their interactions with Acromyrmex leafcutter ants or their gardens. Kaohsiung J Med Sci, 2002 Jan, 18(1), 17 - 22 Monocyte-derived cytokine--IL-12, TGF-beta 1 and TNF-alpha in patients with tuberculosis; Tsai WC et al.; Pulmonary tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (MTB) . This microorganism is capable of inducing a delayed hypersensitivity in the lung, with subsequent expression of the disease . This reaction depends on the presence of different cytokines that exert specific functions . To study the variability of different cytokine responses after MTB antigenic challenge, we used antigens derived from MTB to stimulate the monocytes from both normal healthy contact and the patients with active tuberculosis . We found in the resting state monocytes from healthy contact secreted higher amounts of IL-12 than those from patients . After stimulation with MTB antigen, the secretion of IL-12 did not increase in normal healthy contact, but in patients with tuberculosis the secretion increased . After MTB antigen stimulation, monocytes from patients with active tuberculosis secreted a higher amount of TNF-alpha . In summary, the patterns of monocyte-derived cytokine secretion upon mycobacterial antigen challenge were different in these two groups. Appl Microbiol Biotechnol, 2002 May, 58(6), 823 - 9 Epub 2002 Mar 07. Reconstruction of the biomass history from carbon and nitrogen substrate consumption, ammonia release and proton transfer during solid cultures of Geotrichum candidum and Penicillium camembertii; Aldarf M et al.; Geotrichum candidumand Penicillium camembertii were cultivated on the surface of a gelified medium, simulating the composition of the aqueous phase of a Camembert cheese . The relation of their growth with substrate consumption (carbon or nitrogen), metabolite production (ammonia), or proton transfer (deduced from pH by means of the buffer capacity of the medium) was examined . The coefficients associated with cellular biosynthesis and resulting from cellular maintenance were determined . From these coefficients and the considered substrate utilization or metabolite production kinetics, the growth kinetics were reconstructed until the end of growth . The model allowed analysis of biosynthesis and cellular maintenance contributions to the considered kinetics . At the end of growth, almost all the peptone was used for G . candidum biosynthesis, while most of the lactic acid (62%) was used for cellular maintenance . P . camembertii metabolized fewer amino acids as carbon sources, resulting in use of peptone for maintenance (12%), and lactic acid (80%) for cell biosynthesis . For both microorganisms, ammonia production was growth-associated, since this production resulted from the deamination of carbon- and nitrogen-source amino acids, in close relation with peptone consumption. Med Arh, 2002, 56(2), 89 - 92 {Laboratory diagnosis of Q-fever with the indirect immunofluorescence test}; Beslagic E et al.; Q-Fever nowadays presents the most diffuse disease in the world, caused by the microorganisms from the family Risckettiacae . This disease is Coxiellae burnetii . The laboratory diagnosis of the Q-Fever can be stated either by the isolation of the causer from the patient material, either by the proving of the specifically antibodies . The serologic diagnostics presents the choile method in Q-Fever . The aim of this work is to illustrate the results of the detection of the serum in patients with the clinical symptoms at the infection Coxiellom burnetii . We tested the sera of the patient from the region of the Federation Bosnia and Herzegovina, which arrived in our laboratory in the period of November 2000 till May 2001 . From the total 174 prepared sera specific IgM of the antibodies we found in cases, and the specific IgG of the antibodies in 54 sera. Rhinology, 2002 Mar, 40(1), 24 - 7 Correlation of middle meatus and ethmoid sinus microbiology in patients with chronic sinusitis; Ozcan M et al.; Empirical antibiotics constitute the cornerstone of medical therapy for chronic sinusitis due to difficulties of obtaining cultures from the paranasal sinuses . Indirect isolation of the pathogenic microorganisms outside the paranasal sinuses with a non-invasive method may enable administration of specific antibiotics . In this prospective study, we obtained cultures from the middle meatus and ethmoid sinuses of 193 sides from 127 patients who had undergone FESS for chronic sinusitis with a method that minimizes the risk of nasal contamination . The same bacterial species were isolated from both the ethmoid sinus and middle meatus in 59.3% of the cultures . There was no bacterial growth in either site in 32.3% of the cultures . The overall correlation rate of middle meatus and ethmoid sinus cultures was estimated to be 91.6% . In conclusion, middle meatal cultures can be used for the isolation of pathogenic microorganisms indirectly, while administration of specific antibiotics can be possible according to the results of these cultures. Am J Obstet Gynecol, 2002 May, 186(5 Suppl Understanding), S190 - 7 Nausea and vomiting of pregnancy in an evolutionary perspective; Sherman PW et al.; The proximate mechanisms underlying gestational nausea and vomiting have been intensively studied, but the possibility that the symptoms themselves serve a useful function has only recently been considered seriously . We synthesized evidence to evaluate various hypotheses for the adaptive significance of nausea and vomiting of pregnancy, as well as the possibility that symptoms are nonfunctional byproducts of pregnancy hormones . We found greatest support for the hypothesis that normal levels of nausea and vomiting of pregnancy (excluding hyperemesis) protect pregnant women and their embryos from harmful substances in food, particularly pathogenic microorganisms in meat products and toxins in strong-tasting plants . We discuss the data that support critical predictions of this "maternal and embryo protection hypothesis" (and contradict other hypotheses), as well as appropriate implications of these results . Knowledge that normal nausea and vomiting of pregnancy indicates the functioning of a woman's defense system, rather than a bodily malfunction, may reassure patients and enable health care providers to develop new ways of minimizing the uncomfortable symptoms. Dermatology, 2002, 204 Suppl 1, 42 - 6 Skin disinfection before epidural catheterization: comparative study of povidone-iodine versus chlorhexidine ethanol; Kasuda H et al.; BACKGROUND: Chlorhexidine is better than povidone-iodine for skin preparation before intravascular device insertion or blood culture collection, but it is not known whether chlorhexidine is superior in reducing the colonization of continuous epidural catheters . METHODS: Patients requiring an epidural catheter for postoperative analgesia were randomly assigned to receive skin preparation with 0.5% chlorhexidine ethanol (CE group) or 10% povidone-iodine (PI group) before catheter insertion . Using surgical aseptic techniques, catheters were inserted into either the lumbar or the thoracic epidural space . Gloves used at catheter insertion, swabs of insertion site skin and the catheter tip at catheter removal were qualitatively cultured . RESULTS: Of 70 randomly assigned patients, 62 were evaluable . The clinical characteristics of the patients and the risk factors for infection were similar in the two groups . Catheters were kept in place for 49 +/- 7 h (mean +/- SD) . Seven cultures from gloves yielded microorganisms . In 1 case, the leak test of gloves was positive . Fifteen cultures of catheter insertion sites yielded microorganisms: 7/28 (25%) in the PI group and 8/34 (24%) in the CE group . Six cultures of catheter tips yielded microorganisms: 3/28 (11%) in the PI group and 3/34 (9%) in the CE group . As for the incidence of isolation of bacteria, no difference was seen between the two groups . In none of these 62 cases was any inflammatory reaction seen in the skin locally at catheter removal . CONCLUSIONS: The effect of 0.5% chlorhexidine ethanol is not different from that of 10% povidone-iodine in reducing catheter colonization associated with short-term epidural catheter placement . Proc Natl Acad Sci U S A, 2002 May 14, 99(10), 6556 - 61 Origin of supposedly biogenic magnetite in the Martian meteorite Allan Hills 84001; Barber DJ et al.; Crystals of magnetite (Fe(3)O(4)) and periclase (MgO) in Fe-Mg-Ca carbonate in the Martian meteorite Allan Hills 84001 were studied by using transmission electron microscopy to understand their origin and evaluate claims that the magnetites were made by Martian microorganisms . In magnesian carbonate, periclase occurs as aggregates of crystals (grain size approximately equal to 3 nm) that are preferentially oriented with respect to the carbonate lattice . Larger periclase crystals approximately equal to 50 nm in size are commonly associated with voids of similar size . Periclase clearly formed by precipitation from carbonate as a result of partial decomposition and loss of CO(2) . Magnetite occurs in more ferroan carbonate, and, like periclase, it is associated with voids and microfractures and the two oxides may be intermixed . Magnetite nanocrystals that are commonly euhedral and entirely embedded in carbonate are topotactically oriented with respect to the carbonate lattice, showing that they formed as solid-state precipitates . Magnetites in Fe-rich carbonate rims are not well oriented . These magnetites are generally more irregular in shape and diverse in size than the euhedral variety . All occurrences of magnetite and periclase are entirely consistent with in situ growth by solid-state diffusion as a result of carbonate decomposition during impact heating . Biogenic sources should not be invoked for any magnetites. J Appl Microbiol, 2002, 92(6), 1028 - 34 D,L-Hydantoinase activity of an Ochrobactrum anthropi strain; Pozo C et al.; AIMS: A microorganism with the ability to release methionine from D,L-(2-methylthioethyl) hydantoin (strain 245) was isolated from soil . The aim of this study was the identification of the strain and the adjustment of the conditions of growth and of the enzymatic reaction, in order to achieve high specific activities of bioconversion of the hydantoin . METHODS AND RESULTS: Strain 245 was identified as Ochrobactrum anthropi . The strain grew at alkaline pH (up to 10.0) and its hydantoinase activity was found to be inducible by the substrate D,L-(2-methylthioethyl) hydantoin . The enzyme is also alkalostable, with a pH optimum of 9.0 . Under these conditions, hydantoinase activity was significantly enhanced and its half life prolonged when 200 mmol l-1 ammonium and phosphate were added . The addition of Ca2+, Na+, Cu2+, Co2+, Mg2+, Zn2+ or Fe3+ (0.5 mmol l-1) to the reaction mixture increased the hydantoinase activity of strain 245 up to tenfold after 24 h of incubation, compared with unamended controls . CONCLUSION: The adequate adjustment of some environmental parameters (pH, addition of inducer, presence of ammonium, phosphate, heavy metals and other ions) can considerably increase the D, L-hydantoinase activity of strain 245 . SIGNIFICANCE AND IMPACT OF THE STUDY: The findings reported here set up the initial conditions for a further application of strain 245 in the production of methionine from hydantoine. J Agric Food Chem, 2002 May 22, 50(11), 3236 - 43 Effects of soil pH and soil water content on prosulfuron dissipation; Hultgren RP et al.; The sulfonylurea herbicide prosulfuron, 1-(4-methoxy-6-methyltriazin-2-yl)-3-{2-(3,3,3-trifluoropropyl)phenylsulfonyl}urea, is used for the selective control of broadleaf weeds in corn, sorghum, and cereal grains . To investigate its fate in soils, this study examined the effects of soil pH and water content on the rates of dissipation processes and the products formed under aerobic conditions . Radiometry and chromatography analyses were used to quantify the degradation products and bound residues formed in incubations of 10 different soils . The pH-dependent hydrolysis of the sulfonylurea bridge to form phenyl sulfonamide was the primary transformation process . Significant microbial degradation of prosulfuron occurred in 2 of the 10 soils, yielding (14)CO(2) and desmethyl prosulfuron among the major products . The time required for 50% dissipation of the herbicide (DT(50)) was determined for each soil and water content treatment . At equivalent water contents, prosulfuron DT(50) values were positively correlated with soil pH (P < 0.0001), varying from 6.5 days at pH 5.4 to 122.9 days at pH 7.9 . Soil pH and water content strongly influence the fate of sulfonylurea herbicides in agricultural fields . Differences in the effect of soil water content on dissipation kinetics in a comparison of two soils were attributed to differences in soil pH, texture, and the ability of indigenous microorganisms to transform the herbicide. Metab Eng, 2002 Apr, 4(2), 170 - 81 A general method for determining the contribution of split pathways in metabolite production in the yeast Saccharomyces cerevisiae; Woldman Y et al.; We describe a simple method using (13)C labeling and NMR spectroscopy to determine the flux contributions of alternative pathways in Saccharomyces cerevisiae that produce the same metabolite with identical labeling patterns . Cells were incubated with a (13)C-labeled precursor for one of the branches, and the absolute enrichment of the product and its metabolic precursor(s) was quantified . The ratio of the absolute enrichment of the product to that of its precursor reflects the contribution of the pathway . The method was applied to the biosynthesis of glycine in yeast, which can occur from threonine via threonine aldolase or from serine via serine hydroxymethyltransferase . {2-(13)C}Aspartate and {2-(13)C}serine were used as labeled precursors for the threonine aldolase and serine hydroxymethyltransferase pathways, respectively . The data show that in cells possessing both pathways, the serine hydroxymethyltransferase pathway contributes 65-75% of the total glycine production . In comparison with other approaches, this method provides an inexpensive, flexible alternative to determining the flux contributions of split pathways under controlled conditions and should have wide applicability in the metabolic engineering of microorganisms. FEMS Microbiol Lett, 2002 Apr 9, 209(2), 313 - 9 Detection of methanotrophs with highly divergent pmoA genes from Arctic soils; Pacheco-Oliver M et al.; Tundra soil samples from the Canadian Arctic community, Kuujjuaq, were analyzed for the presence of the soluble (sMMO) and particulate (pMMO) methane monooxygenase genes . Total genomic DNA extracted from these soils was used as template for PCR using sMMO- and pMMO-specific primers, mmoX1-mmoX2 and A189-A682, respectively . pMMO and sMMO genes were detected in the Kuujjuaq soil samples . Isolation of sMMO-possessing methanotrophic microorganisms from the three soils, as determined by the colony naphthalene oxidation assay, was carried out using direct plating (5 degrees C) and methane enrichment studies (5 degrees C and 25 degrees C) . Direct plating did not yield sMMO-possessing methanotrophic bacteria, whereas methane enrichments yielded isolates possessing and expressing sMMO activity . Analysis of derived amino acid sequences of pmoA genes and partial 16S rRNA genes obtained by PCR, using DNA isolated directly from this environment and from isolates, revealed the presence of highly divergent PmoA/AmoA sequences and 16S rRNA sequences that cluster closely with but are distinct from the genes from the genera Methylosinus and Methylocystis. FEMS Microbiol Lett, 2002 Apr 9, 209(2), 295 - 9 Quantitative analysis of phytopathogenic ascomycota on leaves of pedunculate oaks (Quercus robur L.) by real-time PCR; Heuser T et al.; Leaves of oak trees are often infected by various pathogenic fungi . As it is difficult to remove such organisms quantitatively from the leaf surface and as it is often impossible to grow these organisms independently from their host, there are almost no molecular data available from these oak leaf specific pathogens . For the quantitative removal of the microorganisms a procedure was developed combining a wax and microorganism freezing method with a DNA extraction technique . For the development of a species specific detection, DNA of pathogenic filamentous fungi was isolated from hyphae of the upper leaf surface of Quercus robur . Three different species could be identified as (i) Cladosporium sp., (ii) Ramularia sp . and (iii) Microsphaera alphitoides by amplifying and sequencing an 18S-28S segment of their rDNA . For the final quantification a real-time PCR protocol was established allowing the species specific quantification of the three pathogenic filamentous fungi . The whole procedure was successfully applied to quantify the amount of the three species on oak leaves collected in autumn. FEMS Microbiol Rev, 2002 Mar, 26(1), 73 - 81 Microbial carboxyl esterases: classification, properties and application in biocatalysis; Bornscheuer UT; Esterases (EC 3.1.1.x) represent a diverse group of hydrolases catalyzing the cleavage and formation of ester bonds and are widely distributed in animals, plants and microorganisms . Beside lipases, a considerable number of microbial carboxyl esterases have also been discovered and overexpressed . This review summarizes their properties and classification . Special emphasis is given on their application in organic synthesis for the resolution of racemates and prostereogenic compounds . In addition, recent results for altering their properties by directed evolution are presented. J Electron Microsc (Tokyo), 2002, 51(2), 87 - 91 Study of NADPH oxidase-activated sites in human neutrophils; Seguchi H et al.; Neutrophils represent the first line of defence against invading microorganisms . An important part of this defence mechanism is the generation of superoxide (O2*-) and its reactive derivatives after stimulation by a variety of agents . This oxidant production is linked to the activation of NADPH oxidase, which is composed of cytosolic components (p47-phox and p67-phox) and membrane components (p22-phox and gp91-phox) . Previous studies have shown that NADPH oxidase resides in the plasma membrane and the traditional view holds that cytoplasmic components of NADPH oxidase are brought into the neighbourhood of the plasma membrane and then conjugated with its membrane components upon stimulation . This review focuses on the evaluation of NADPH oxidase-activated sites in human neutrophils . Based on electron microscopic analysis, O2*- is first generated upon stimulation with certain stimulants, such as phorbol myristate acetate, within a specialized intracellular compartment containing alkaline phosphatase, and not on the plasma membrane, as previously thought . In addition, the cytosolic component of NADPH oxidase, p47-phox, accumulates at the juxtaposition of intracellular compartments but not of the plasma membrane . These results demonstrate that initial O2*- production occurs in an intracellular pool in human neutrophils . The oxidant-producing granules then bind directly to the plasma membrane or fuse to form larger structures that eventually become to be associated with the plasma membrane, and O2*- is released extracellularly from the neutrophils. Science, 2002 May 10, 296(5570), 1071 - 7 Geomicrobiology: how molecular-scale interactions underpin biogeochemical systems; Newman DK et al.; Microorganisms populate every habitable environment on Earth and, through their metabolic activity, affect the chemistry and physical properties of their surroundings . They have done this for billions of years . Over the past decade, genetic, biochemical, and genomic approaches have allowed us to document the diversity of microbial life in geologic systems without cultivation, as well as to begin to elucidate their function . With expansion of culture-independent analyses of microbial communities, it will be possible to quantify gene activity at the species level . Genome-enabled biogeochemical modeling may provide an opportunity to determine how communities function, and how they shape and are shaped by their environments. Science, 2002 May 10, 296(5570), 1068 - 71 Microbial behavior in a heterogeneous world; Fenchel T; Most microorganisms are motile during at least part of their life cycle, because they need to find optimal conditions in a patchy world . The sheer volume of microorganisms in the biosphere means that their motile sensory behavior also contributes to the global transformation and cycling of matter . How microorganisms move and how they orient themselves using environmental cues are integral to understanding the complex structure and function of microbial communities, but although motility in response to external stimuli was first described more than 120 years ago, understanding of the cellular and molecular mechanisms involved has only been achieved more recently. Waste Manag, 2002, 22(2), 215 - 28 Waste ecocompatibility in storage and reuse scenarios: global methodology and detailed presentation of the impact study on the recipient environments; Perrodi Y et al.; In 1995, the ADEME launched a research program called "Waste Ecocompatibility" in order to define a reliable methodology for measuring the impact of waste in storage or reuse scenarios . The French concept of "Ecocompatibility" is defined as the situation where the pollutant flux from waste disposed of or used in specified conditions is compatible with the environmental acceptance of the receiving environments . The chief feature of this definition is to integrate the evaluation of the three following terms: pollutants emission from the waste, transport of the pollutants from the waste to the receptor cells and the environmental acceptance of the receiving environments . The "Waste Ecocompatibility" program consisted of a literature survey and an experimental part . The literature study aimed to determine factors and waste characteristics to be considered for a reliable ecocompatility assessment, to provide an overview of the available tools for measuring those factors and characteristics and to propose a first approach of the methodology . In the framework of the experimental program, this approach was then applied to three theoretical scenarios to validate the laboratory tools (comparative study of laboratory and field results) and to calibrate the global methodology . This paper deals with the results of the experimental program concerning the impact study on receiving environments: impact on plants and microorganisms living in soil, impacts on soil fauna and aquatic fauna . In other papers we intend to present the operational methodology for the assessment of waste ecocompatibility . It includes bio-assays at laboratory scale (microcosms), pilot scale (mesocosms) and in situ experiments (experimental prairie) . To limit the use of in situ experiments other research works are necessary to validate bio-assays at laboratory or pilot scale. Crit Rev Microbiol, 2002, 28(1), 1 - 26 Emerging waterborne infections: contributing factors, agents, and detection tools; Theron J et al.; Because microorganisms are easily dispersed, display physiological diversity, and tolerate extreme conditions, they are ubiquitous and may contaminate and grow in water . The presence of waterborne enteric pathogens (bacteria, viruses, and protozoa) in domestic water supplies represents a potentially significant human health risk . Even though major outbreaks of waterborne disease are comparatively rare, there is substantial evidence that human enteric pathogens that are frequently present in domestic water supplies are responsible for low-level incidence of waterborne microbial disease . Although these diseases are rarely debilitating to healthy adults for more than a few hours to a few days, enteric pathogens can cause severe illness, even death, for young children, the elderly, or those with compromised immune systems . As the epidemiology of waterborne diseases is changing, there is a growing global public health concern about new and reemerging infectious diseases that are occurring through a complex interaction of social, economic, evolutionary, and ecological factors . New microbial pathogens have emerged, and some have spread worldwide . Alternative testing strategies for waterborne diseases should significantly improve the ability to detect and control the causative pathogenic agents . In this article, we provide an overview of the current state of knowledge of waterborne microbial pathogens, their detection, and the future of new methods in controlling these infectious agents. J Med Assoc Thai, 2001 Dec, 84 Suppl 3, S650 - 7 Chronic infections and atherosclerosis; Leowattana W; The established risk factors for atherosclerosis such as hypertension, smoking, diabetes mellitus, hyperlipidemia, and hyperhomocysteinemia do not explain clinical and epidemiological features of coronary heart disease (CHD) . The role of infectious disease as a CHD risk factor may partly explain these features . Chronic infection with various microorganisms, particularly, Chlamydia pneumoniae, Cytomegalovirus (CMV) and Helicobacter pylori may play a role in etiological factors, linking inflammation and atherogenesis . Results from epidemiological studies, pathology of atherosclerotic plaques, animal studies, molecular biology and clinical antibiotic trials indicated a positive association between C . pneumoniae infection and CHD . Chronic infection might also influence preexisting plaque by enhancing T cell activation, which participate in destabilization of intimal cap . However, the exact nature of pathophysiological link between the organisms and CHD remains to be elucidated . Future antibiotic interventional studies may help to further clarify the role of chronic infection and inflammation in CHD. J Appl Microbiol, 2002, 92 Suppl, 72S - 7S Products containing biocides: perceptions and realities; Favero MS; The mechanisms of action for chemical germicides and antibiotics for inactivating microorganisms are significantly different and methods for determining resistance by microorganisms to these agents are also different . Chemical germicides usually have multiple targets and the mechanisms for inactivation and resistance are not measured in absolute terms but rather in the rapidity with which they reduce levels of microorganisms . The term tolerance is much more suited for germicides than the term resistance . The mechanism of resistance to chemical germicides is often dependent on the concentration of the germicide . At high concentrations multiple cellular and metabolic targets are involved, and at low concentrations fewer cellular targets . In contrast antibiotics usually have a singular cellular or metabolic target and resistance implies the ability of the microorganism to grow in the presence of the antibiotic, and in a clinical sense, to initiate or continue infection in the presence of the antibiotic . When methods used to assess resistance to antibiotics are applied to chemical germicides, inappropriate interpretations can be made regarding the ability of microorganisms to develop resistance to antibiotics as a result of developing resistance to chemical germicides . The use of chemical germicides in health-care institutions and especially the home setting has increased in recent years . Although there may be an overuse of germicides in these settings the consequence is a cost issue and not one that involves the development of antibiotic resistant microorganisms. J Contam Hydrol, 2002 Mar, 55(1-2), 161 - 74 Microbially mediated redox processes in natural analogues for radioactive waste; Haveman SA et al.; Natural analogues allow scientists to investigate biogeochemical processes relevant to radioactive waste disposal that occur on time scales longer than those that may be studied by time-limited laboratory experiments . The Palmottu U-Th deposit in Finland and the Bangombe natural nuclear reactor in Gabon involve the study of natural uranium, and are both considered natural analogues for subsurface radioactive waste disposal . The microbial population naturally present in groundwater may affect the redox conditions, and hence, the radionuclide solubility and migration . Therefore, groundwater samples from the two sites were investigated for microbial populations . The total numbers of cells ranged from 10(4) to 10(6) cells ml(-1) . Iron-reducing bacteria (IRB) were the largest culturable microbial population in the Palmottu groundwater and were present at up to 1.3 x 10(5) cells ml(-1) . Sulfate-reducing bacteria (SRB) and acetogens could also be cultured from the Palmottu groundwater . The numbers of IRB and SRB were largest in groundwater with the lowest uranium concentrations . Removal of dissolved U(VI) from solution was concomitant with the growth of IRB enrichment cultures and the reduction of iron . The redox buffer in the Palmottu groundwater consists of iron and uranium species, both of which are affected by IRB . IRB and aerobic heterotrophs were cultured from the Bangombe groundwater, where redox potentials are buffered by iron and organic carbon species . Microbial populations similar to those found at Palmottu and Bangombe are found throughout the Fennoscandian Shield, a potential host rock for subsurface radioactive waste disposal . These results confirm that microorganisms can be expected to play a role in stabilizing radioactive waste disposed of in the subsurface by lowering redox potential and immobilizing radionuclides. Free Radic Res, 2002 Jan, 36(1), 33 - 45 N-acetylcysteine improves in vitro the function of macrophages from mice with endotoxin-induced oxidative stress; Victor VM et al.; Reactive oxygen species (ROS) and proinflammatory cytokines produced by immune cells cause the oxidative stress involved in septic shock induced by endotoxin . This oxidative stress can be controlled to a certain degree by antioxidants, which is specially important for a type of immune cell, i.e . the phagocyte, that uses ROS to kill microorganisms and needs antioxidants in order to support its functions . In a previous study we have observed changes in several functions of peritoneal macrophages from BALB/c mice with lethal endotoxic shock caused by intraperitoneal injection of Escherichia coli lipopolysaccharide (LPS) (100 mg/kg), which were associated with a high production of superoxide anion . N-acetylcysteine (NAC) is a thiolic antioxidant that improves the immune response, and we have observed that when administered intraperitoneally (150 mg/kg) at 30 min after LPS injection it counteracts the effects of LPS on macrophages and lymphocytes . In the present work, we have studied the in vitro effect of several concentrations of NAC (0.001, 0.01, 0.1, 1 and 2.5 mM) on the following functions: adherence to substrate, chemotaxis, ingestion of particles, ROS production and the release of tumor necrosis factor (TNFalpha) of peritoneal macrophages from BALB/c mice at 2, 4,12 and 24 h after LPS injection . The results show that the administration of NAC (especially at 0.1 mM) decreases raised adherence, ingestion, ROS production and TNFalpha levels in macrophages from animals injected with LPS, bringing these functions to values near those of control animals . These effects which seem to be linked to a modulation of NF-kappaB, suggest that the improvement of immune functions observed in previous work after injection of NAC to animals with endotoxic shock could be due to a direct action of this thiol antioxidant on immune cells. Immunobiology, 2002 Mar, 205(1), 120 - 31 Immunoregulatory activities of lactoferrin in the delayed type hypersensitivity in mice are mediated by a receptor with affinity to mannose; Zimecki M et al.; The mannose receptor (MR) mediates clearance of pathogenic microorganisms and potentially harmful glycoproteins by recognition of the carbohydrate structures . This manuscript describes the significance of the MR in mediation of the lactoferrin (LF) effects in delayed type hypersensitivity (DTH) in mice . Here we demonstrate that the inhibitory effect of LF on the effector phase of DTH to sheep red blood cells (SRBC) was abolished by intraperitoneal (i.p.) pretreatment of mice with 50 mg of methyl-alpha-D-mannopyranoside (MMan) . Also, we report that the inhibition of the effector DTH response to ovalbumin (OVA) was reversed by MMan but not by D-galactose (Gal) . MMan was found to reduce the adjuvant activity of LF given together with OVA, however, did not interfere with the upstimulatory effect of the complete Freund's adjuvant . Comparative studies between bovine and human lactoferrins (BLF and HLF), revealed that the adjuvant effect of BLF was stronger than that of HLF, nevertheless, both effects were inhibited by MMan . Thus, we postulate, that MR is a primary receptor for lactoferrin in mediation of its some immunotropic activities. Atherosclerosis, 2002 Jun, 162(2), 253 - 9 Chlamydial infection in canine atherosclerotic lesions; Sako T et al.; We attempted to detect chlamydial antigens in canine atherosclerotic lesions from seven dogs by immunohistochemical technique using anti-Chlamydia psittaci (C . psittaci) polyclonal and anti-C . pneumoniae monoclonal antibodies . Immunopositive signals to both antibodies were recognized in the atherosclerotic lesions of the aortas, coronary and splenic arteries of all dogs . Positive signals were found in the foamy cytoplasm of infiltrated macrophages and extracellular matrices in the lesions . In some lesions, cytoplasm of the endothelial cells and smooth muscle cells was also immunopositive against both antibodies . By electron microscopy, chlamydial microorganisms were found in the cytoplasm of endothelial cells and smooth muscle cells . Using polymerase chain reaction (PCR), detection of C . pneumoniae DNAs were performed in the spleen, heart (coronary arteries) and kidney in one of the seven dogs . Positive 314 bp PCR products were obtained in all samples of the dog . These results confirmed the presence of viable Chlamydiae in atheromas and supported the conclusion that the organism may be an active factor in the pathogenesis of canine, as well as human atherosclerosis. J Toxicol Environ Health A, 2002 May 10, 65(9), 685 - 93 Furosemide and spironolactone reduce transmigration of leukocytes through endothelial cell monolayers; Hofbauer R et al.; Furosemide and spironolactone reduce transmigration of leukocytes through endothelial cell monolayers . Leukocytes play a tremendous role during inflammation . Leukocytes migrate from intravascular space into the tissue to attack microorganisms . Various agents are able to influence leukocyte recruitment . The influence of diuretics, such as furosemide and spironolactone, on inflammatory processes is not well known . The aim of our study was to examine the influence of furosemide and spironolactone on leukocyte migration through endothelial cell monolayers (ECM) . Human umbilical vein endothelial cells were cultured on microporous membranes achieving a monolayer . Polymorphonuclear leukocytes (PMNL) were used in a currently described migration assay . PMNL and/or ECM were pretreated with furosemide and spironolactone using therapeutic, as well as higher and lower, concentrations . Furosemide (76 +/- 7.2%) and spironolactone (70 +/- 7.7%) were able to inhibit PMNL migration through ECM significantly, when both cell types were treated simulating the situation after an iv injection . Furosemide and spironolactone were identified as potent inhibitors of leukocyte migration through ECM. Environ Sci Technol, 2002 Apr 15, 36(8), 1809 - 14 The application of tris buffer and copper sulfate for the preservation of phenylurea pesticides analyzed using U.S . EPA method 532 in the UCMR Survey; Bassett MV et al.; A high performance liquid chromatographic method was developed to meetthe U.S . Environmental Protection Agency's (EPA) Unregulated Contaminant Monitoring Rule (UCMR) Survey need for the analysis of phenylurea pesticides in drinking waters . Many of these phenylurea compounds were demonstrated to degrade rapidly in the presence of the residual chlorine disinfectant in drinking waters . This degradation was halted by the addition of a tris buffer, which was initially chosen to optimize the sample pH prior to extraction . Copper sulfate was found to prevent the regrowth of microorganisms in surface waters, which was observed upon dechlorination . Tris buffer provided the additional benefit of keeping the copper sulfate preservative in solution even in groundwater matrices that caused precipitation of copper in its absence . A C18 solid phase, in cartridge or disk form, was used to efficiently extract target compounds from the preserved drinking water matrices . A 21-day storage stability study, together with precision and accuracy studies, demonstrated thatthis method had suitable sensitivity, selectivity, accuracy, precision, and ruggedness for use in the EPA's UCMR drinking water occurrence survey. Adv Biochem Eng Biotechnol, 2002, 74, 65 - 98 The molecular mechanism of ATP synthesis by F1F0-ATP synthase: a scrutiny of the major possibilities; Nath S; A critical goal of metabolism in living cells is the synthesis of adenosine triphosphate (ATP) . ATP is synthesized by the enzyme F1F0-ATP synthase . This enzyme, the smallest-known molecular machine, couples proton translocation through its membrane-embedded, hydrophobic domain, F0, to the synthesis of ATP from adenosine diphosphate (ADP) and inorganic phosphate (Pi) in its soluble, hydrophilic headpiece, F1 . Animals, plants and microorganisms all capture and utilize energy by this important chemical reaction . How does it occur? The binding change mechanism and the torsional mechanism of energy transduction and ATP synthesis are two mechanisms that have been proposed in the literature . According to the binding change mechanism (which considers reversible catalysis and site-site cooperativity), energy is required primarily for release of synthesized ATP, but not for its synthesis . On the other hand, according to the torsional mechanism (which considers an irreversible mode of catalysis and absence of cooperativity), all the elementary steps require energy, and the ion-protein interaction energy obtained from the ion gradients is used to synthesize ATP, for Pi binding, and for straining the beta-epsilon bond in order to enable ADP to bind . The energy to release preformed ATP from the tight catalytic site (betaDP) is provided by the formation of the beta-epsilon ester linkage . First, the central features of these mechanisms are clearly delineated . Then, a critical scrutiny of these mechanisms is undertaken . The predictions of the torsional mechanism are listed . In particular, how the torsional mechanism deals with the specific difficulties associated with other mechanisms, and how it seeks to explain a wealth of structural, spectroscopic, and biochemical data is discussed in detail . Recent experimental data in support of the mechanism are presented . Finally, in view of the molecular machine nature of energy transduction, the indispensability of applying engineering tools at the molecular level is highlighted . This paves the way for the development of a new field: Molecular Physiological Engineering. Adv Biochem Eng Biotechnol, 2002, 74, 207 - 38 Integrated approach to explore the potential of marine microorganisms for the production of bioactive metabolites; Wagner-Dobler I et al.; During the last 10 years marine organisms have provided a large number of new natural products . Interesting compounds have mainly been derived from macroorganisms such as sponges, ascidians, corals and bryozoans . The number of secondary metabolites from marine microorganisms is smaller, but rapidly increasing . Because of the enormous difficulties involved in harvesting products from marine animals, and the fact that some of the bioactive compounds are produced by associated bacteria, the advantages of sustainable production of bioactive metabolites by bacteria or fungi, under the protection of natural resources, seem to be very attractive for the future . This review describes current progress in the isolation and identification of novel marine microorganisms, the discovery of new secondary metabolites, the biotechnological approaches to overproduce them, as well as the evaluation and characterization of their bioactivity. Biochem Cell Biol, 2002, 80(2), 189 - 95 Production and characterization of an in vitro engineered human oral mucosa; Rouabhia M et al.; The role of epithelial cells in oral pathologies is poorly understood . Until now, most studies have used normal or transformed epithelial cell monolayers, a system that largely bypasses oral mucosal complexity . To overcome these limitations, an engineered human oral mucosa (EHOM) model has been produced and characterized . Following histological and immunohistochemical analyses, EHOM showed well-organized and stratified tissues in which epithelial cells expressed proliferating keratins such as Ki-67, K14, and K19 and also differentiating keratin (K10) . In this model, epithelial cells interacted with fibroblasts in the lamina propria by secreting basement membrane proteins (laminins) and by expressing integrins (beta1 and alpha2beta1) . Cytokine analyses using cultured supernatants showed that cells in EHOM were able to secrete interleukins (IL) including IL-1beta and IL-8 and tumor necrosis factor alpha (TNF-alpha) . Finally, cells in this engineered model were able to secrete different metalloproteinases such as gelatinase-A and gelatinase-B . In conclusion, using tissue engineering technology, we produced well-organized EHOM tissues . It is anticipated that this model will be useful for examining mechanisms involved in oral diseases under controlled conditions by modeling the interactions between mucosa and microorganisms in the oral cavity. Nature, 2002 May 23, 417(6887), 411 - 8 Epub 2002 May 01. Adaptation in the chemotactic guidance of nerve growth cones; Ming GL et al.; Pathfinding by growing axons in the developing nervous system may be guided by gradients of extracellular guidance factors . Analogous to the process of chemotaxis in microorganisms, we found that axonal growth cones of cultured Xenopus spinal neurons exhibit adaptation during chemotactic migration, undergoing consecutive phases of desensitization and resensitization in the presence of increasing basal concentrations of the guidance factor netrin-1 or brain-derived neurotrophic factor . The desensitization is specific to the guidance factor and is accompanied by a reduction of Ca2+ signalling, whereas resensitization requires activation of mitogen-associated protein kinase and local protein synthesis . Such adaptive behaviour allows the growth cone to re-adjust its sensitivity over a wide range of concentrations of the guidance factor, an essential feature for long-range chemotaxis. Microb Ecol, 2002 Jan, 43(1), 134 - 45 Epub 2001 Jan 23. Specific 16S rDNA sequences associated with naphthalene degradation under sulfate-reducing conditions in harbor sediments; Hayes LA et al.; Previous studies have demonstrated that naphthalene and other polycyclic aromatic hydrocarbons (PAHs) can be anaerobically oxidized with the reduction of sulfate in PAH-contaminated marine harbor sediments, including those in San Diego Bay . In order to learn more about the microorganisms that might be involved in anaerobic naphthalene degradation, the microorganisms associated with naphthalene degradation in San Diego Bay sediments were evaluated . A dilution-to-extinction enrichment culture strategy, designed to recover the most numerous culturable naphthalene-degrading sulfate reducers, resulted in the enrichment of microorganisms with 16S rDNA sequences in the d-Proteobacteria, which were closely related to a previously described pure culture of a naphthalene-degrading sulfate reducer, NaphS2, isolated from sediments in Germany . A more traditional enrichment culture approach, expected to enrich for the fastest-growing naphthalene-degrading sulfate reducers, yielded 16S rDNA sequences closely related to those found in the dilution-to-extinction enrichments and NaphS2 . Analysis of 16S rDNA sequences in sediments from two sites in San Diego Bay that had been adapted for rapid naphthalene degradation by continual amendment with low levels of naphthalene suggested that the microbial community composition in the amended sediments differed from that present in the unamended sediments from the same sites . Most significantly, 6-8% of the sequences recovered from 100 clones of each of the naphthalene-amended sediments were closely related to the 16S rDNA sequences in the enrichment cultures as well as the sequence of the pure culture, NaphS2 . No sequences in this NaphS2 phylotype were recovered from the sediments that were not continually exposed to naphthalene . A PCR primer, which was designed based on these phylotype sequences, was used to amplify additional 16S rDNA sequences belonging to the NaphS2 phylotype from PAH-degrading sediments from Island End River (Boston), MA, and Liepaja Harbor, Latvia . Closely related sequences were also recovered from highly contaminated sediment from Tampa Bay, FL . These results suggest that microorganisms closely related to NaphS2 might be involved in naphthalene degradation in harbor sediments . This finding contrasts with the frequent observation that the environmentally relevant microorganisms cannot be readily recovered in pure culture and suggests that further study of the physiology of NaphS2 may provide insights into factors controlling the rate and extent of naphthalene degradation in marine harbor sediments. Med Oral, 2002 May-Jun, 7(3), 206 - 21 Quantitative and functional neutrophil deficiencies; Diz-Dios P et al.; Neutrophil is a cell involved in the maintenance of homeostasis of the organism . It plays an important role in the elimination and control of certain pathogenic microorganisms . Deficiency in function and quantity of neutrophils, either due to congenital or secondary etiological extrinsic factors, lead to recurrent infectious processes of variable severity . The most frequent oral complications in these patients are ulcers, periodontal disease and candidiasis . Due to their high susceptibility to infections and the risk implied in undergoing surgical procedures, actions leading to a higher degree of prophylaxis must be carried out on these patients (oral hygiene habits, fluoridation, dietary advise, etc.) . The use of granulocyte colony-stimulating factors (G-CSF) allows, in some cases, the fullfillment of some, until recently considered controversial dental procedures . Its administration demands an adequate selection of patients and a close collaboration between physician and dental practitioner . Despite G-CSF effectiveness, dental treatment in severe neutropenic patients must be carried out in a hospital and, in some cases, regardless of adequate care, the progress of periodontal disease cannot be avoided. Proc Natl Acad Sci U S A, 2002 May 14, 99(10), 6901 - 6 Epub 2002 Apr 30. Global changes in gene expression by human polymorphonuclear leukocytes during receptor-mediated phagocytosis: cell fate is regulated at the level of gene expression; Kobayashi SD et al.; Phagocytes are a critical component of the innate immune response in humans and eliminate invading microorganisms through a process known as phagocytosis . Two distinct receptor-linked phagocytic pathways, one with Ab receptors (FcRs; FcR, Fc receptor) and the other complement receptors (CRs), mediate binding and ingestion of pathogens by human polymorphonuclear leukocytes (PMNs) . Although progress has been made toward defining complex signal transduction processes that underlie phagocytosis in each pathway, very little is known about gene regulation during or after phagocytosis . Therefore, we used human oligonucleotide microarrays to identify changes in expression of 12,561 genes accompanying FcR- and CR-mediated phagocytosis . Eighty-four percent of 279 differentially expressed genes were induced or repressed 90 min after ingestion of Ab- and/or complement-opsonized particles . Unexpectedly, more than 30 of these genes encoded proteins involved in at least three distinct apoptotic pathways . Ninety-four differentially expressed cell fate-related genes were identified between 180 and 360 min after phagocytosis and most were induced or repressed by PMNs activated through both receptors simultaneously . By using flow cytometry, we found that FcR- and CR-mediated phagocytosis each promoted programmed cell death in human PMNs; however, phagocytosis mediated by the combination of FcRs and CRs induced apoptosis earlier than that by either receptor alone . Our results reveal distinct patterns of receptor-mediated gene expression that define complex inducible apoptotic pathways in activated PMNs . Most significantly, we discovered that programmed cell death is regulated at the level of gene expression . Thus, we hypothesize that gene regulation in PMNs facilitates resolution of inflammatory responses. Biochim Biophys Acta, 2002 Mar 25, 1595(1-2), 392 - 6 Exploring hyperthermophilic proteins under pressure: theoretical aspects and experimental findings; Mombelli E et al.; Proteins from hyperthermophilic microorganisms are generally capable of withstanding temperatures close to, or even higher than the boiling point . As a rule, these proteins are strongly piezostable as well, although exceptions have been also reported . This observation has a theoretical relevance, as the understanding of the effects of pressure and temperature on protein stability is equally important to develop a comprehensive model for their thermodynamic stability . Nevertheless, the structural features justifying the correlation between heat resistance and pressure resistance are poorly understood . Actually, most reports do not exceed the phenomenological level . Only in the case of the small protein Sso7d from Sulfolobus solfataricus, characterisation of wild-type and some mutants showed that both properties are largely accounted for by a network of aromatic residues found in the hydrophobic core of the molecule . Current knowledge, however, does not allow to establish to what extent this finding may be generalised . In a biotechnological perspective, hyperthermophilic enzymes seem to be more suitable for bioprocesses at high pressure with respect to their mesophilic counterparts . Indeed, thanks to their higher resistance towards pressure and temperature, they may be exploited in a much broader range of working conditions for tuning activity and specificity . Furthermore, they are often activated by increasing pressure, although it cannot be established, to date, to what extent this is a common feature. Biochim Biophys Acta, 2002 Mar 25, 1595(1-2), 367 - 81 Pressure effects on in vivo microbial processes; Bartlett DH; Pressures between 10 and 100 MPa can exert powerful effects on the growth and viability of organisms . Here I describe the effects of elevated pressure in this range on mesophilic (atmospheric pressure adapted) and piezophilic (high-pressure adapted) microorganisms . Examination of pressure effects on mesophiles makes use of this unique physical parameter to aid in the characterization of fundamental cellular processes, while in the case of piezophiles it provides information on the essence of the adaptation of life to high-pressure environments, which comprise the bulk of our biosphere . Research is presented on the isolation of pressure-resistant mutants, high-pressure regulation of gene expression, the role of membrane lipids and proteins in determining growth ability at high pressure, pressure effects on DNA replication and topology as well as on cell division, and the role of extrinsic factors in modulating enzyme activity at high pressure. Transfus Med, 2002 Apr, 12(2), 85 - 106 Tick-borne diseases in transfusion medicine; Pantanowitz L et al.; Ticks are effective vectors of viral, bacterial, rickettsial and parasitic diseases . Many of the tick-borne diseases (TBDs) are of significance to transfusion medicine, either because of the risks they pose to the blood supply or the necessity for blood products required in their treatment . The transmission of tick-borne pathogens via blood transfusion is of global concern . However, among transfusion medicine practitioners, experience with most of these microorganisms is limited . Transfusion transmission of TBDs has been documented largely by means of single case reports . A better understanding of the epidemiology, biology and management of this group of diseases is necessary in order to assess the risks they pose to the blood supply and to help guide effective prevention strategies to reduce this risk . Unique methods are required to focus on donor selection, predonation questioning, mass screening and inactivation or eradication procedures . The role of the transfusion medicine service in their treatment also needs to be better defined . This article reviews the growing body of literature pertaining to this emerging field of transfusion medicine and offers some recommendations for transfusionists in dealing with TBDs. Ther Apher, 2002 Apr, 6(2), 116 - 8 Adsorption of endotoxin by beta2-microglobulin adsorbent column (Lixelle): the new approach for endotoxinemia; Tsuchida K et al.; We previously reported that Lixelle, which was used for beta2-microglobulin (BMG) adsorption columns, could adsorb not only BMG but also inflammatory cytokines . We became interested in the application of Lixelle for patients with endotoxinemia and researched its ability to adsorb microorganism components in vitro using lipopolysaccharide (LPS) (Escherichia coli B8), endotoxin (ET) contaminated water . The initial concentrations of each water solution were LPS (ET 29,135 EU/L) and contaminated water (ET 3,523 EU/L) whole blood solution was LPS (ET 1,197.6 EU/L) . Each 2.5 ml of the stock solution and adjusted diluted solutions contained 0.5 ml of Lixelle beads . After shaking at 37 degrees C for 2 h, ET in the solutions was determined using the endotoxin specific-limulus amebocyte lysate method . The results revealed that even though ET concentrations in LPS and contaminated water incubated in water solution and in whole blood were high, the samples containing Lixelle beads showed significant decreases . Thus, Lixelle beads can adsorb not only BMG but also microorganism components such as LPS and ET . These findings together with the ability of Lixelle to adsorb ET show the possibility of the application for treatment of endotoxinemia. J Agric Food Chem, 2002 May 8, 50(10), 2731 - 41 Phytoalexins from the Vitaceae: biosynthesis, phytoalexin gene expression in transgenic plants, antifungal activity, and metabolism; Jeandet P et al.; Resistance of plants to infection by phytopathogenic microorganisms is the result of multiple defense reactions comprising both constitutive and inducible barriers . In grapevine, the most frequently observed and best characterized defense mechanisms are the accumulation of phytoalexins and the synthesis of PR-proteins . Particular attention has been given here to stilbene phytoalexins produced by Vitaceae, specifically, their pathway of biosynthesis (including stilbene phytoalexin gene transfer experiments to other plants) and their biological activity together with fungal metabolism. Med Clin North Am, 2002 Mar, 86(2), 219 - 38 Prevention of tick-borne diseases; Wilson ME; Ticks are a part of the landscape where humans live, work, and play . Because ticks carry a wide range of organisms that potentially can cause disease in humans, many studies have focused on ways to reduce risk of these diseases . Ticks have biologically complex interactions with microorganisms and with their vertebrate hosts, on whom they depend for blood meals and survival . To consider ways to reduce the burden of tick-borne diseases in humans, it is necessary to understand the biology and ecology of ticks and their interface with humans . In many areas, changes in land use, reforestation, and patterns of human settlements have led to more abundant tick populations, increasing rates of infections in ticks, and increasing contact with human populations . Warmer winter temperatures in temperate regions may extend the transmission season for some ticks and pathogens . Although much of the discussion in this article has focused on I . scapularis and the Lyme disease spirochete (because they have been studied extensively), other tick-pathogen pairs may differ in risk factors for infection and transmission dynamics . Interventions studied to reduce the burden of tick-borne diseases include changing the environment, controlling vertebrate hosts, killing ticks, altering the behavior of humans, treating tick bites, and trying to protect humans through immunologic means (vaccine) . All of these approaches have limitations and drawbacks . From a public health perspective, a plan that employs multiple strategies may be most effective . This article has reviewed what is known about preventive interventions, including the vaccine. Life Sci Space Res, 1968, 6, 108 - 14 The survival of terrestrial microorganisms in space at orbital altitudes during Gemini satellite experiments; Hotchin J et al.; In a previous series of rocket- and satellite-borne experiments, microorganisms were exposed to space between altitudes of 60 to 460 km for periods between 3 minutes and 4 months . The results showed that some of the unprotected microorganisms survived the direct exposure for up to 17 hours . Complete survival was found when the microorganisms were shielded from nonpenetrating radiation . These results made it desirable to systematically study the lethal influence of the space environment on a wider spectrum of unprotected microorganisms over various exposure periods . This communication presents the results of exposure experiments on board the Gemini XII satellite . The microorganisms were dried on plastic coated aluminum plates using techniques which will be described elsewhere . During the flight of the Gemini XII space capsule, the microorganisms were directly exposed to space for approximately 6 1/2 hours . After the successful completion of the Gemini XII mission the payload was returned to the laboratory for elution and titration of the microorganisms using techniques described elsewhere . The results showed that again survival of some microorganisms had occurred . An attempt is made to integrate these results with data obtained in previously published experiments. Life Sci Space Res, 1968, 6, 100 - 7 The biological effectiveness of solar electromagnetic radiation in space; Lorenz PR et al.; An attempt has been made to determine the solar ultraviolet action spectrum for dried coliphage T-1 exposed to space under various filters at sounding rocket altitudes between 80 to 150 km . The survival of these microorganisms was compared with the preliminary results of laboratory ultraviolet irradiation experiments using monochromatic light sources of wavelengths of 1633 angstroms, 2062 angstroms, 2537 angstroms and of 3064 angstroms to 3200 angstroms . The results indicate that ultraviolet light of wave-lengths between 2000 angstroms to 3000 angstroms was mainly responsible for the killing of the filter protected microorganisms . More studies of the biological action spectrum of the vacuum ultraviolet light including the Lyman alpha emission are needed to determine the cause of the more than 100 fold higher death rate of the microorganisms which were not protected during the space exposure by filters. Int J Food Microbiol, 2002 Apr 5, 74(3), 203 - 16 Osmoregulation and its importance to food-borne microorganisms; O'Byrne CP et al.; The control of water activity has been used as a means of preserving foods for thousands of years . This preservation strategy presents food-borne microorganisms with serious problems, many of which relate to the management of water flow . Although the specific details of how each organism deals with these problems are different, several common themes have emerged . Bacteria induce specific responses . both physiological and genetic, to respond to either the loss or the gain of water, triggered by changes in the osmolarity of the environment . Many of the key systems have now been identified and the mechanisms of their regulation are beginning to be understood . Here we review recent developments in the field of bacterial osmoregulation with emphasis on key food-borne genera. Electrophoresis, 2002 Apr, 23(7-8), 1070 - 9 Adaption of a fragment analysis technique to an automated high-throughput multicapillary electrophoresis device for the precise qualitative and quantitative characterization of microbial communities; Trotha R et al.; The analysis of microbial communities is of increasing importance in life sciences and bioengineering . Traditional techniques of investigations like culture or cloning methods suffer from many disadvantages . They are unable to give a complete qualitative and quantitative view of the total amount of microorganisms themselves, their interactions among each other and with their environment . Obviously, the determination of static or dynamic balances among microorganisms is of fast growing interest . The generation of species specific and fluorescently labeled 16S ribosomal DNA (rDNA) fragments by the terminal restriction fragment length polymorphism (T-RFLP) technique is a suitable tool to overcome the problems other methods have . For the separation of these fragments polyacrylamide gel sequencers are preferred as compared to capillary sequencers using linear polymers until now because of their higher electrophoretic resolution and therefore sizing accuracy . But modern capillary sequencers, especially multicapillary sequencers, offer an advanced grade of automation and an increased throughput necessary for the investigation of complex communities in long-time studies . Therefore, we adapted a T-RFLP technique to an automated high-throughput multicapillary electrophoresis device (ABI 3100 Genetic Analysis) with regard to a precise qualitative and quantitative characterization of microbial communities. Clin Infect Dis, 2002 May 15, 34(10), 1362 - 8 Epub 2002 Apr 17. The promise of novel technology for the prevention of intravascular device-related bloodstream infection . II . Long-term devices; Crnich CJ et al.; Intravascular devices (IVDs) are widely used for vascular access but are associated with a substantial risk of IVD-related bloodstream infection (BSI) . The development of novel technologies based on our understanding of pathogenesis promises a quantum reduction in IVD-related infections in an era of growing nursing shortage . Infections of long-term IVDs (most are in place for > or =10 days), including cuffed and tunneled central venous catheters (CVCs), implanted subcutaneous central venous ports, and peripherally inserted central catheters (PICCs), are primarily due to microorganisms that gain access to the catheter hub and lumen . Novel securement devices and antibiotic lock solutions have been shown to reduce the risk of IVD-related BSI in prospective randomized trials . The challenge for the future will be to identify new preventative technologies and to begin to more-widely adapt those technologies that have already been shown to be efficacious and cost effective. J Clin Microbiol, 2002 May, 40(5), 1621 - 5 18S ribosomal DNA typing and tracking of Acanthamoeba species isolates from corneal scrape specimens, contact lenses, lens cases, and home water supplies of Acanthamoeba keratitis patients in Hong Kong; Booton GC et al.; We examined partial 18S ribosomal DNA (Rns) sequences of Acanthamoeba isolates cultured in a study of microbial keratitis in Hong Kong . Sequence differences were sufficient to distinguish closely related strains and were used to examine links between strains obtained from corneal scrape specimens, contact lenses, lens cases, lens case solutions, and home water-supply faucets of patients with Acanthamoeba . We also looked for evidence of mixed infections . Identification of Acanthamoeba Rns genotypes was based on sequences of approximately 113 bp within the genus-specific amplicon ASA.S1 . This permitted genotype identification by using nonaxenic cultures . Of 13 specimens obtained from corneal scrapes, contact lenses, lens cases, or lens case solutions, 12 were Rns genotype T4 and the remaining one was Rns genotype T3 . The sequences of corneal scrape specimens of two patients also were the same as those obtained from their contact lenses or lens case specimens . A possible triple-strain infection was indicated by three different T4 sequences in cultures from one patient's lenses . Although faucet water used by patients to clean their lenses is a possible source of infections, specimens isolated from the faucets at two Acanthamoeba keratitis patients' homes differed from their corneal scrape or lens specimens . The overall results demonstrate the potential of this Rns region for tracking Acanthamoeba keratitis strains in infections and for distinguishing single-strain and closely related multiple-strain infections even when other microorganisms might be present with the cultured specimens . They also confirm the predominance of Rns genotype T4 strains in Acanthamoeba keratitis infections. Folia Microbiol (Praha), 2002, 47(1), 56 - 60 Quantitative distribution of microbial biomass in the soil profile of a high-mountain grassy ecosystem; Bogoev VM et al.; The soil microbiota of a grassy ecosystem in the subalpine belt of the Rila Mountain National Park showed greater amounts of fungal biomass . This remained relatively constant throughout the months of sampling while bacterial biomass was a dynamic value fluctuating within a wide range . The two groups of microorganisms also differed in their in-depth distribution in the soil profile: the bacterial biomass was relatively homogeneously distributed while the fungal biomass gradually decreased with depth . Good correlation between the amount of biomass and the values of some abiotic factors of the environment was shown by correlation analysis in a "warm" period of investigation; no distinct correlation between microbial biomass and environmental factors was observed in a "cold" period. Inflamm Bowel Dis, 2002 May, 8(3), 213 - 8 Role of infections in the manifestation or reactivation of inflammatory bowel diseases; Stallmach A et al.; The etiology of inflammatory bowel disease (IBD) is unknown . In addition to genetic and environmental factors, microorganisms have been discussed as possibly playing an important role . Recent reports in the literature do not suggest that a specific persistent infection causes IBD, but indicate that enteric pathogens could cause initial onset of IBD and are associated with reactivation of quiescent disease . Despite their self-limited character, these infections initiate a cascade of inflammatory events leading to chronic, relapsing disease in a genetically susceptible host ("hit-and-run" hypothesis) . Epidemiological and microbiologic studies suggest that enteropathogenic microorganisms play a substantial role in the clinical initiation and relapses of IBD . However, similar to traveler's diarrhea, the frequency of infections in the first manifestation and in relapses of IBD is probably understated, due to the problems in detecting enteric pathogens . Thus microbiologic screening is helpful in patients with flares of IBD for optimal medical treatment. Free Radic Biol Med, 2002 May 1, 32(9), 790 - 6 Carbonyl modified proteins in cellular regulation, aging, and disease; Levine RL; The oxidative modification of proteins by reactive species is implicated in the etiology or progression of a panoply of disorders and diseases . The level of these modified molecules can be quantitated by measurement of the protein carbonyl content, which has been shown to increase in a variety of diseases and processes, notably during aging . For the most part, oxidatively modified proteins are not repaired and must be removed by proteolytic degradation, a process which normally proceeds very efficiently, from microorganisms to mammals . In eukaryotes, removal is usually carried out by the proteosome, which selectively degrades oxidatively modified proteins, whether they be damaged by reactive oxygen species or specifically oxidized by cellular regulatory processes . The molecular deficiencies that cause accumulation of oxidatively modified proteins are not identified, but regardless of cause, the accumulation is likely to disrupt normal cellular function. Acta Crystallogr D Biol Crystallogr, 2002 May, 58(Pt 5), 861 - 3 Epub 2002 Apr 26. Crystallization and preliminary X-ray crystallographic analysis of aspartate 1-decarboxylase from Helicobacter pylori; Kwon AR et al.; Aspartate 1-decarboxylase (PanD) catalyzes the alpha-decarboxylation of L-aspartate in the major route of beta-alanine production for pantothenate biosynthesis in bacteria . Pantothenate is synthesized in microorganisms, plants and fungi but not in animals and thus the enzymes of its biosynthetic pathway are potential targets for developing agents against these organisms . PanD from the pathogenic bacterium Helicobacter pylori has been overexpressed in Escherichia coli and crystallized using sodium formate as a precipitant . Crystals diffracted to better than 1.5 A Bragg spacing upon exposure to synchrotron X-rays . Diffraction data to 1.55 A have been collected from a crystal grown in the presence of the substrate analogue isoasparagine . The crystal belongs to the tetragonal space group I422, with unit-cell parameters a = b = 81.83, c = 93.78 A . The asymmetric unit contains one subunit of PanD, with a corresponding crystal volume per protein mass (V(M)) of 2.85 A(3) Da(-1) and a solvent content of 56.8%. Appl Environ Microbiol, 2002 May, 68(5), 2610 - 3 Thermodynamic analysis of growth temperature dependence in the adhesion of Candida parapsilosis to polystyrene; Gallardo-Moreno AM et al.; The purpose of this work was to study the adhesion to polystyrene of two Candida parapsilosis strains, grown at 22 and 37 degrees C, in terms of hydrophobicity, surface charge, and interaction free energy . Growth temperature changed the surface properties of microorganisms, yielding a good correlation between thermodynamic predictions and adhesion behavior. Appl Environ Microbiol, 2002 May, 68(5), 2605 - 9 Fate of Escherichia coli O157:H7 in manure-amended soil; Jiang X et al.; Escherichia coli O157:H7 cells survived for up to 77, >226, and 231 days in manure-amended autoclaved soil held at 5, 15, and 21 degrees C, respectively . Pathogen populations declined more rapidly in manure-amended unautoclaved soil under the same conditions, likely due to antagonistic interactions with indigenous soil microorganisms . E . coli O157:H7 cells were inactivated more rapidly in both autoclaved and unautoclaved soils amended with manure at a ratio of 1 part manure to 10 parts soil at 15 and 21 degrees C than in soil samples containing dilute amounts of manure . The manure-to-soil ratio, soil temperature, and indigenous microorganisms of the soil appear to be contributory factors to the pathogen's survival in manure-amended soil. Heart Dis, 2001 Jan-Feb, 3(1), 14 - 7 Effects of garlic extract (Allium sativum) on neutrophil migration at the cellular level; Hofbauer R et al.; Studies of the effects of garlic extract on oxidative and lipoprotein levels have yielded widely different findings . Leukocytes play an important role during many processes, including inflammation . They migrate from intravascular spaces into tissues and attack microorganisms . In a recent study, the inhibitory effects of the nonsteroidal antiinflammatory drug, ibuprofen, on leukocyte transmigration were demonstrated using an in vitro assay . Little is known about the cellular effects of garlic extracts (Allium sativum) . The aim of the current study was to investigate the influence of garlic extract on leukocyte migration through endothelial cell monolayers and thereby evaluate a possible role in inflammatory processes . Human umbilical endothelial cells were cultured on microporous membranes to make an endothelial cell monolayer (ECM) . Freshly isolated neutrophils were used in a recently described migration assay . The amount of untreated neutrophils migrating through the untreated ECM was used as control and set at 100% . Neutrophils and/or ECM were pretreated with garlic extract using moderate, higher, and lower concentrations . Moderate plasma concentrations of garlic extract inhibited neutrophil migration through ECM significantly (64 +/- 5.8% standard deviation {SD}; P < 0.05) when both cell types were treated, (a situation that may have clinical relevance) . Treating either neutrophils or ECM alone showed significant reductions in migratory rate (neutrophils treated alone: 81 +/- 7.7% SD, P < 0.05; ECM alone: 70 +/- 6.7%, P < 0.05) . Thus, garlic extract is identified as a potent inhibitor of leukocyte migration through endothelial cell monolayers . Treatment of both cell types has an additive effect . Endothelial cells seem to be more affected than neutrophils . Further investigations are necessary to understand the potential clinical consequences. Pathologe, 2002 Jan, 23(1), 20 - 37 {Histological diagnosis of inflammatory skin diseases . Use of a simple algorithm and modern diagnostic methods}; Hugel H; The histological diagnosis of inflammatory skin diseases on a day-to-day routine basis poses the difficult task to characterise a dynamic clinical process by histomorphological analysis of one single lesion . Very complex algorithms which are meant to lead to the correct diagnosis are difficult to use . To facilitate the process and reduce the number of algorithms the following method is proposed: 1 . Histological examination under low power Definition of lesions altered by scratching 2 . Localisation of the significant pathological alterations as follows: Changes in epidermis and dermis Blister formation Changes in dermis/subcutis without characteristic changes in the epidermis Changes mainly in the subcutis 3 . Closer examination using a simple algorithm and planning of further investigations following defined criteria: In the case of changes in epidermis and dermis definition of spongiotic, psoriasiform or lichenoid dermatitis (Abb . 1) . In the case of blister formation definition of the blister following the given algorithm (Abb . 1) . In the case of changes in dermis/subcutis without characteristic changes in the epidermis after exclusion of vasculitis and lymphoma definition of the main pattern as lymphocytic, neutrophilic, eosinophilic, lymphoplasmocytic or granulomatous and following the given algorithm (Abb . 2) . In the case of changes mainly in the subcutis after exclusion of vasculitis and lymphoma definition of the main pattern of panniculitis as septal or/and lobular (Abb . 2) . Exclusion of PAS positive microorganisms and checking if the general pattern fits into infectious correlation . Use of clinicopathological correlation. J Immunol, 2002 May 1, 168(9), 4553 - 8 Dominant effects of mutations in the collagenous domain of mannose-binding protein; Wallis R; Individuals heterozygous for mutant alleles encoding serum mannose-binding protein (MBP, also known as mannose-binding lectin) show increased susceptibility to infections caused by a wide range of pathogenic microorganisms . To investigate the molecular defects associated with heterozygosity, wild-type rat serum MBP polypeptides (MBP-A: 56% identical in sequence to human MBP) and rat MBP polypeptides containing mutations associated with human immunodeficiency have been coexpressed using a well-characterized mammalian expression system . The resulting proteins are secreted almost exclusively as heterooligomers that are defective in activating the complement cascade . Functional defects are caused by structural changes to the N-terminal collagenous and cysteine-rich domains of MBP, disrupting interactions with associated serine proteases . The dominant effects of the mutations demonstrate how the presence of a single mutant allele gives rise to the molecular defects that lead to the disease phenotype in heterozygous individuals. Antioxid Redox Signal, 2002 Feb, 4(1), 49 - 60 Assembly and activation of the neutrophil NADPH oxidase in granule membranes; Karlsson A et al.; Phagocyte activation is accompanied by assembly of an NADPH oxidase that reduces oxygen to form a number of reactive species . These oxygen radicals can eradicate invading microorganisms, regulate the function of other immune reactive cells, and cause damage to "innocent bystander" cells . It is generally assumed that the NADPH oxidase is activated exclusively in the plasma membrane . In neutrophils, this assumption does not fit with the subcellular localization of the membrane component of the oxidase, which is stored in granule compartments . It has now become increasingly evident that oxidants are also produced in an intracellular compartment that we identify as the specific granules . Myeloperoxidase is stored in another granule subset, the azurophil granules, and participates in the processing of the oxidative metabolites . We suggest that neutrophil activation is accompanied by fusion between azurophil and specific granules, allowing these peroxidase-dependent reactions to take place . The presented data suggest a requisite role for neutrophil oxidants complementing their function as microbial killing agents . Signaling capabilities of the oxidants, affecting for example, the state of protein phosphorylation, regulation of transcription factors, and induction of apoptosis, are discussed.
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