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| Methods Cell Sci, 2001, 23(4), 175 - 84 Comparing qualitative and quantitative spectroscopic techniques for the detection of the effect of direct iron loading of mammalian cell cultures; Traore HN et al.; Iron overload augments diseases of the liver and microorganism infection as well as deregulates the immune system . In vitro analysis of the effects of iron loading and its chelation involves determining the amount of iron constituting overload, which metal sources and cell lines to use and reliable assay methods . The uptake of 500 microM FeSO4 or FeEDTA by CEMss, U937 or leukocytes was confirmed by inductively coupled plasma-atomic emission spectroscopy (ICP-AES) . Excess iron increased CEMss viability (assessed by MTT, XTT, Trypan- and Alamar Blue) by an average of 18% (P = 0.034) . Flow cytometry indicated dye-viable cells to be undergoing apotosis/necrosis while still confirming an increase (9%, P < 0.001) in excess iron-induced viability . The iron chelator desferioxamine (DFO) when added in addition to Fe, reversed the effects of excess iron (and vice versa) and had detrimental effects when used on its own (33% inhibition of viability as measured by dyes and 10.85%; P = 0.0427 assessed by flow cytometry) . The 4 dyes demonstrated different levels of sensitivity in detecting the influence of iron or DFO but gave a related, qualitative picture while flow cytometry and ICP-AES data was more quantitative. J Bacteriol, 2003 Jan, 185(1), 285 - 94 Common extracellular sensory domains in transmembrane receptors for diverse signal transduction pathways in bacteria and archaea; Zhulin IB et al.; Transmembrane receptors in microorganisms, such as sensory histidine kinases and methyl-accepting chemotaxis proteins, are molecular devices for monitoring environmental changes . We report here that sensory domain sharing is widespread among different classes of transmembrane receptors . We have identified two novel conserved extracellular sensory domains, named CHASE2 and CHASE3, that are found in at least four classes of transmembrane receptors: histidine kinases, adenylate cyclases, predicted diguanylate cyclases, and either serine/threonine protein kinases (CHASE2) or methyl-accepting chemotaxis proteins (CHASE3) . Three other extracellular sensory domains were shared by at least two different classes of transmembrane receptors: histidine kinases and either diguanylate cyclases, adenylate cyclases, or phosphodiesterases . These observations suggest that microorganisms use similar conserved domains to sense similar environmental signals and transmit this information via different signal transduction pathways to different regulatory circuits: transcriptional regulation (histidine kinases), chemotaxis (methyl-accepting proteins), catabolite repression (adenylate cyclases), and modulation of enzyme activity (diguanylate cyclases and phosphodiesterases) . The variety of signaling pathways using the CHASE-type domains indicates that these domains sense some critically important extracellular signals. Ecotoxicol Environ Saf, 2002 Nov, 53(3), 416 - 21 INT-dehydrogenase activity test for assessing anaerobic biodegradability of organic compounds; Hongwei Y et al.; This study assessed anaerobic biodegradability of organic compounds from microorganism activity . Dehydrogenase activity can be a good parameter characterizing the microorganism activity . A modified method of 2-(p-iodophenyl-3-(p-nitrophenyl)-5-pheny tetrazolium chloride-dehydrogenase activity determination was proposed in anaerobic biodegradability assessment . Cubic spline curves were adopted to link the data points . This curve was integrated twice to calculate areas . The microorganism activity index in anaerobic biodegradability assessment was calculated by standardizing the integral . According to the results of the activity index, 14 kinds of organic compounds were classified into readily, partially, and poorly biodegradable under anaerobic conditions, respectively . As a result, some conclusions for anaerobic biodegradability of organic compounds were reached, based on the activity index value. Pediatr Allergy Immunol, 2002 Dec, 13(6), 438 - 42 Immunoglobulin G antibodies of children exposed to microorganisms in a water-damaged school; Savilahti R et al.; The aim of this study was to determine whether exposure to fungi (molds and yeasts) among children attending a water-damaged school was reflected by the children's immunoglobulin G (IgG) response to microorganisms typical of water damage and whether the presence of these IgG antibodies was associated with respiratory symptoms and morbidity . The relationships between positive IgG antibodies and atopy, described as elevated allergen-specific immunoglobulin E (IgE) antibodies, were also examined . The study population consisted of a randomly selected group of exposed children attending a water-damaged school and a group of unexposed children of the same age . Serum samples for analyses of IgG and IgE antibodies were drawn from the children . The respiratory morbidity, the number of positive IgG antibodies to nine microorganisms indicating water damage, and IgE sensitization to common environmental allergens (Phadiatop) were studied . The mean number of positive IgG findings was significantly higher among the exposed children . The number of positive IgG antibodies did not correlate with respiratory illnesses or symptoms at the individual level even though the exposed children who had positive IgG antibodies to four or more microorganisms in the total group comparison tended to have higher respiratory morbidity . In the exposed group, a negative correlation was found between the number of positive IgG antibodies and the total value of allergen-specific IgE antibodies . As among adults exposed to microorganisms at work, IgG antibodies in children seem to be a relevant indicator of exposure to microorganisms in a water-damaged school on the group level. Helicobacter, 2002 Dec, 7(6), 378 - 83 A group-specific PCR assay for the detection of Helicobacteraceae in human gut; Bohr UR et al.; BACKGROUND: Enterohepatic Helicobacter species are emerging pathogens, which are increasingly isolated from humans with enteric diseases . Nevertheless, current methods to detect Helicobacteraceae in the human gut have significant limitations . METHODS: Based on 16S-rRNA gene alignments and computer aided primer analysis a set of group-specific PCR primers was designed . The evaluation of the PCR assay was performed using 36 ATCC reference strains and intestinal biopsies from 10 patients with defined gastric Helicobacter pylori status . The amplification products derived from clinical samples were cloned and subsequently analyzed by DNA sequencing . Sensitivity of the PCR-assay was determined by spiking previously negative tested samples with decreasing amounts of Helicobacter DNA . RESULTS: The analysis of the ATCC reference strains revealed amplification products in all 14 Helicobacter strains and Wolinella succinogenes, 21 other microorganisms representing negative controls did not produce PCR fragments . Four out of the 10 patient-derived samples were positive . Three of them represented H . pylori-derived DNA confirming the gastric H . pylori infection in these patients . In the fourth patient, who was suffering from Crohn's disease, H . pullorum was identified . The sensitivity of the PCR assay was 0.1 pg of Helicobacter-derived DNA representing about 40 bacteria . CONCLUSION: The novel PCR assay described here is an important new tool in rapid and sensitive assessment for the presence of Helicobacteraceae in human gut. J Gen Appl Microbiol, 2000 Dec, 46(6), 283 - 296 Biochemical and genetic bases of microbial degradation of polychlorinated biphenyls (PCBs); Furukawa K; The microbial degradation of polychlorinated biphenyls (PCBs) has been extensively conducted by many workers, and the following general results have been obtained . (1) PCBs are degraded oxidatively by aerobic bacteria and other microorganisms such as white rot fungi . PCBs are also reductively dehalogenated by anaerobic microbial consortia . (2) The biodegradability of PCBs is highly dependent on chlorine substitution, i.e., number and position of chlorine . The degradation and dehalogenation capabilities are also highly strain dependent . (3) Biphenyl-utilizing bacteria can cometabolize many PCB congeners to chlorobenzoates by biphenl-catabolic enzymes . (4) Enzymes involved in the PCB degradation were purified and characterized . Biphenyl dioxygenase, ring-cleavage dioxygenase, and hydrolase are crystallized, and two ring-cleavage dioxygenases are being solved by x-ray crystallography . (5) The bph gene clusters responsible for PCB degradation are cloned from a variety of bacterial strains . The structure and function are analyzed with respect to the evolutionary relationship . (6) The molecular engineering of biphenyl dioxygenases is successfully performed by DNA shuffling, domain exchange, and subunit exchange . The evolved enzymes exhibit wide and enhanced degradation capacities for PCBs and other aromatic compounds. Proc Natl Acad Sci U S A, 2002 Dec 24, 99(26), 17037 - 42 Epub 2002 Dec 13. Discovery of sulfated metabolites in mycobacteria with a genetic and mass spectrometric approach; Mougous JD et al.; The study of the metabolome presents numerous challenges, first among them being the cataloging of its constituents . A step in this direction will be the development of tools to identify metabolites that share common structural features . The importance of sulfated molecules in cell-cell communication motivated us to develop a rapid two-step method for identifying these metabolites in microorganisms, particularly in pathogenic mycobacteria . Sulfurcontaining molecules were initially identified by mass spectral analysis of cell extracts from bacteria labeled metabolically with a stable sulfur isotope (34SO 4 2-) . To differentiate sulfated from reduced-sulfur-containing molecules, we employed a mutant lacking the reductive branch of the sulfate assimilation pathway . In these sulfur auxotrophs, heavy sulfate is channeled exclusively into sulfated metabolites . The method was applied to the discovery of several new sulfated molecules in Mycobacterium tuberculosis and Mycobacterium smegmatis . Because a sulfur auxotrophic strain is the only requirement of the approach, many microorganisms can be studied in this manner . Such genetic engineering in combination with stable isotopic labeling can be applied to various metabolic pathways and their products. J Pharmacol Toxicol Methods, 2001 Sep-Oct, 46(2), 93 - 101 Permeability measurement of macromolecules and assessment of mucosal antigen sampling using in vitro converted M cells; Liang E et al.; INTRODUCTION: M cells are located in the epithelial layer covering the gut-associated lymphoid tissue and are responsible for delivery of macromolecules and microorganisms to the underlying lymphoid cells . It has been shown that the human colonic cell line Caco-2 can be converted to M cells in vitro following coculture with isolated lymphocytes from murine Peyer's patches . Studies were undertaken to evaluate and characterize the transepithelial transport of select macromolecules across these in vitro derived M cells . METHODS: Caco-2 cells were converted to M cells as reported previously . The morphology of Caco-2 cells and M cells was compared by transmission electron microscopy (TEM) . The transport properties of macromolecules such as horseradish peroxidase, FITC-conjugated polystyrene beads, and radiolabeled dextrans were examined . The activation of murine antigen-specific T cells following transport of the antigen ovalbumin across the M-cell barrier was assessed by measuring cytokine production . RESULTS: M cells were shown to be irregular in shape and have fewer and shorter microvilli compared to the Caco-2 cell progenitors . These cells were still able to form tight junctions and monolayers on polycarbonate membranes . Time-course studies demonstrated that the transport of polystyrene beads and large-molecular-weight dextrans at physiological temperature across M-cell-containing monolayers was size dependent and more rapid than across Caco-2 cell monolayers . The transport of dextrans was also shown to be temperature and concentration dependent . Befitting the role of the M cell in mucosal defense, protein antigen could be delivered by these cells in order to be processed and presented to antigen-specific CD4+ T lymphocytes . DISCUSSION: The M-cell permeability model is a functional and practical system for evaluating the transport properties of macromolecules and assessing the potential for intestinal mucosal antigen sampling to elicit immunological responses. Symp Ser Soc Appl Microbiol, 2002, (31), 72S - 77S 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. Boll Chim Farm, 2002 Sep-Oct, 141(5), 367 - 71 Comparative study between culture media employed in sterility test; Bugno A et al.; The sterility test aims at the contact of a product with a culture medium, as a way of detecting the possible presence of viable microorganisms in products which have been submitted to a sterilization process and/or to aseptic processing . Since the official introduction of the methodology, in 1932, several culture media have been proposed and adopted, in a constant attempt to offer conditions which support the growth of as many contaminants as possible . This work aimed at a comparative evaluation of the efficiency of microbial contaminants detection in the different types of culture media employed in sterility tests . The study led to the conclusion that the culture media recommended by the pharmacopoeia compendia, soybean casein digest and fluid thioglycollate, present the best results in microorganisms detection . Besides, the microbial strains, as well as microbial suspension density, recommended by the main pharmacopoeias to verify culture media growth promoting capacity, have also proved suitable for use. Electrophoresis, 2002 Nov, 23(22-23), 3847 - 57 Recent advances in capillary isoelectric focusing: 1997-2001; Shimura K; The methodological developments in the field of capillary isoelectric focusing (CIEF) published between 1997-2001 are reviewed as a continuation of the previous review by Rodriguez-Diaz et al . (Electrophoresis 1997, 18, 2134-2144) . The applications are summarized and the progress in CIEF technologies, including experimental setup with coated and uncoated capillaries, remedies for the presence of salts in samples, additives to reduce precipitation of samples during the focusing process, calibration of the pH gradients, issues of reproducibility, carrier ampholyte-free CIEF, and a computer simulation of focusing process are discussed . Developments of IEF separations in fabricated microchannels and the advances in detection schemes, i.e., imaging, fluorescence and chemiluminescence, are summarized . The progress in micropreparation was noted, and the massive works for two-dimensional separations are described for the coupling with size-exclusion chromatography and mass spectrometry, in which the developments aimed at proteomics are discussed separately . The applications for the detection of noncovalent complexes and the separations of microorganisms are reviewed. Plant Physiol, 2002 Dec, 130(4), 2177 - 87 The elicitor cryptogein blocks glucose transport in tobacco cells; Bourque S et al.; Cryptogein is a 10-kD protein secreted by the oomycete Phytophthora cryptogea that induces a hypersensitive response on tobacco (Nicotiana tabacum var . Xanthi) plants and a systemic acquired resistance against various pathogens . The mode of action of this elicitor has been studied using tobacco cell suspensions . Our previous data indicated that within minutes, cryptogein signaling involves various events including changes in ion fluxes, protein phosphorylation, sugar metabolism, and, eventually, cell death . These results suggested that transport of sugars could be affected and, thus, involved in the complex relationships between plant and microorganisms via elicitors . This led us to investigate the effects of cryptogein on glucose (Glc) uptake and mitochondrial activity in tobacco cells . Cryptogein induces an immediate inhibition of Glc uptake, which is not attributable to plasma membrane (PM) depolarization . Conversely, cryptogein-induced valine uptake is because of PM depolarization . Inhibition of the PM Glc transporter(s) was shown to be mediated by a calcium-dependent phosphorylation process, and is independent of active oxygen species production . This inhibition was associated with a strong decrease in O(2) uptake rate by cells and a large mitochondrial membrane depolarization . Thus, inhibition of Glc uptake accompanied by inhibition of phosphorylative oxidation may participate in hypersensitive cell death . These results are discussed in the context of competition between plants and microorganisms for apoplastic sugars. FEMS Microbiol Lett, 2002 Dec 17, 217(2), 213 - 7 Models of 2,4,6-trinitrotoluene (TNT) initial conversion by yeasts; Zaripov SA et al.; The original models of the initial steps of 2,4,6-trinitrotoluene (TNT) conversion by yeasts are presented . Saccharomyces sp . ZS-A1 reduced nitro groups of TNT producing isomeric monohydroxylaminodinitrotoluenes (HADNT) as the key initial metabolites (molar ratio HADNT/TNT was up to 0.81), whereas aminodinitrotoluenes (ADNT) and the hydride-Meisenheimer complex of TNT (H-TNT) were the minor products . Conversely, Candida sp . AN-L13 transformed TNT almost quantitatively into H-TNT, thus realizing the alternative attack, consisting of the TNT aromatic ring reduction . The third type of conversion, revealed in Candida sp . AN-L14, is the combination of both above mechanisms and produces an equimolar mix of HADNT and H-TNT . In the toxicity tests with Paramecium caudatum, the supernatant of Saccharomyces sp . ZS-A1, which converts TNT into HADNT, was most toxic while the supernatant of Candida sp . AN-L13 (TNT --> H-TNT) was least toxic . The microorganisms converting TNT quantitatively to the reactive metabolites can be useful for their immobilization through the detoxifying interaction with the soil components such as humic compounds . Scand J Infect Dis, 2002, 34(10), 772 - 4 Severe Legionella pneumophila pneumonia in a patient with iron overload; Vikram HR et al.; Iron plays a crucial role in the energy metabolism of microorganisms . Humans have developed iron-withholding mechanisms as a form of non-specific immunity . We describe a patient with iron overload and severe Legionella pneumophila pneumonia . This report emphasizes the importance of early consideration of and appropriate therapy against Legionella for patients with iron overload who present with community-acquired pneumonia. Curr Drug Targets Immune Endocr Metabol Disord, 2001 Nov, 1(3), 199 - 208 Helicobacter pylori infection, immune response and vaccination; Lembo A et al.; It is well known that abnormal immune responses may play a pathogenic role in the H . pylori-related gastropathy . Indeed, as far as humoral immune response is concerned, it is still debated whether specific anti-H . pylori antibodies have a protective or noxious effect in infected hosts . Besides proinflammatory cytokines released from macrophages, such as tumor-necrosis factor-a and interleukin-1beta, and IFN-gamma derived from T-helper 1 lymphocytes, also interleukin-10, a product of T-helper 2 lymphocytes with antiinflammatory properties, seems to be surprisingly involved in the pathogenesis of H . pylori-induced gastritis . In addition, lipopolysaccharide derived from the outher membrane of H . pylori acts as a chemoattractant for monocytes and induces release of free radicals, interleukin-1beta, interleukin-6, interleukin-8 and tumor necrosis factor-alpha . On the other hand, H . pylori lipopolysaccharide could be responsible for the increased polyamine concentrations in the gastric mucosa and polyamines, such as putrescine, spermidine and spermine, could be involved in the increased cell proliferation and consequent possible neoplastic transformation of the gastric mucosa . Incubation of peripheral blood mononuclear cells with H . pylori increases significantly the surface expression of CD95 receptor (Fas), thus suggesting that these bacteria are able to induce apoptosis . In animal models, different types of vaccination have been investigated, including stimulation of nasal and rectal lymphoid tissue, as well as adoptive transfer of T cell from donors immunized with H . pylori . However, results obtained are frequently disappointing . In humans, urease of H . pylori was safely used as oral vaccine in the absence or presence of adjuvants with encouraging results . Finally, DNA vaccines could offer in the future advantages for prophylactic H . pylori eradication, especially where population is infected by this microorganism since childhood. Gig Sanit, 2002 Sep-Oct, (5), 27 - 8 {Sanitary-toxicological assessment of petroleum slimers undergone biological purification}; Tsinberg MB et al.; Pilot production and production studies using a biological method for treating and neutralizing toxic waste polluted with petroleum products at high concentrations (up to 30%) were conducted . The results have indicated that the use of oil degrading microorganisms is highly promising . Within the first vegetative period as high as 80% of petroleum products withdrew from waste and over 95% of those migrated into the aqueous medium . Purification decreased the content of mobile phases of metals to the values below the maximum allowable concentrations . The waste that is highly toxic for protozoa and the test crop wheat shall be considered to be nontoxic. Gig Sanit, 2002 Sep-Oct, (5), 24 - 6 {Lysozyme of hydrobionts and its role in self-purification of water reservoirs}; Solovykh GN et al.; Model experiments were carried out to study the effects of hydrobiont lysozyme on self-purifying processes of water reservoirs through its participation in the formation of a community of destructive microorganisms . Lysozyme-resistant forms have been found to lie at basis of bacterial cenosis of the Urals, among which there are lysozyme-active and anti-lysozyme-active ones, their population shows an inverse relationship . The exogenous lysozyme that enters the water reservoir in the period when there is a change in the dominant forms of hydrobionts affects the composition of bacterial cenosis by preserving anti-lysozyme-activity with a high activity (6-8 (g/ml), which causes a change in the microbial community of water reservoir, thus affecting the processes of its self-purification . The bacteria having ALA, which do maintain the capacity of a water reservoir for self-clearance from organic substance, persist under the action of endogenous lysozyme of phyto- and zooplankton among saprophytic microorganisms that are destructive agents in the biocenosis. J Biol Chem, 2003 Feb 28, 278(9), 7540 - 52 Epub 2002 Dec 09. Carbohydrate-induced differential gene expression patterns in the hyperthermophilic bacterium Thermotoga maritima; Chhabra SR et al.; The hyperthermophilic bacterium Thermotoga maritima MSB8 was grown on a variety of carbohydrates to determine the influence of carbon and energy source on differential gene expression . Despite the fact that T . maritima has been phylogenetically characterized as a primitive microorganism from an evolutionary perspective, results here suggest that it has versatile and discriminating mechanisms for regulating and effecting complex carbohydrate utilization . Growth of T . maritima on monosaccharides was found to be slower than growth on polysaccharides, although growth to cell densities of 10(8) to 10(9) cells/ml was observed on all carbohydrates tested . Differential expression of genes encoding carbohydrate-active proteins encoded in the T . maritima genome was followed using a targeted cDNA microarray in conjunction with mixed model statistical analysis . Coordinated regulation of genes responding to specific carbohydrates was noted . Although glucose generally repressed expression of all glycoside hydrolase genes, other sugars induced or repressed these genes to varying extents . Expression profiles of most endo-acting glycoside hydrolase genes correlated well with their reported biochemical properties, although exo-acting glycoside hydrolase genes displayed less specific expression patterns . Genes encoding selected putative ABC sugar transporters were found to respond to specific carbohydrates, and in some cases putative oligopeptide transporter genes were also found to respond to specific sugar substrates . Several genes encoding putative transcriptional regulators were expressed during growth on specific sugars, thus suggesting functional assignments . The transcriptional response of T . maritima to specific carbohydrate growth substrates indicated that sugar backbone- and linkage-specific regulatory networks are operational in this organism during the uptake and utilization of carbohydrate substrates . Furthermore, the wide ranging collection of such networks in T . maritima suggests that this organism is capable of adapting to a variety of growth environments containing carbohydrate growth substrates. Hematology, 2002 Dec, 7(6), 359 - 69 Thrombus formation with rehydrated, lyophilized platelets; Fischer TH et al.; Stored human platelets are frequently used in hemorrhagic emergencies, but have limited immediate utility for controlling bleeding due to storage lesion and are frequently contaminated with microorganisms . The development of paraformaldehyde-treated, lyophilized and rehydrated (RL) platelets, which are sterile and have a prolonged shelf life (years), ameliorate the efficacy and sterility problems with stored platelets . RL platelets have been shown to have many native functions of fresh platelets in vitro and to mediate hemostasis in vivo in large animal models of hemorrhagic shock and cardiopulmonary bypass induced platelet dysfunction . To further evaluate the functional properties of this transfusion product, we studied the role of RL platelets in three aspects of thrombus formation and lysis . First, the interaction between RL platelets and fibrinogen was investigated . The surface density of unligated GPIIb-IIIa on RL and fresh platelets were, respectively 30000 and 70000 molecules per cell as detected with the monoclonal antibody 10E-5 . Freezing, lyophilization and rehydration steps in the preparation of RL platelets resulted in the surface presentation of 120000 molecules of fibrinogen per cell from alpha granule sources . After ADP activation, RL platelets bound exogenous 125I-labeled fibrinogen in a dose-dependent manner with an affinity that is similar to that of fresh platelets and was inhibited by RGD peptides . 125I-Labeled fibrinogen binding to RL and fresh platelets, respectively, saturated at 14000 and 32000 molecules per cell . Scanning electron microscopic ultrastructural analysis showed that fibrin strands interacted with the surface of RL platelets in a normal manner . The second set of studies investigated the ability of RL platelets to catalyze and amplify the clot formation process in an activation-dependent manner . We showed that RL platelets undergo degranulation in fibrin in clots and functioned as thrombogenic surfaces for the generation of activated coagulation factors and fibrin generation . A final set of studies was performed to investigate fibrin of clots that contained RL platelets . RL platelet clots were lysed in the presence of tissue plasminogen activator with a similar time course as clots without platelets, and lysis occurred faster than when fresh platelets were included in the fibrin mass . The results of these three studies demonstrate that RL platelets are capable of mediating thrombus formation and do not inhibit lysis . Our results help explain how RL platelets restore hemostasis in vivo, and indicate that these cells might be a viable alternative to fresh stored platelets in transfusion medicine. Medicina (Kaunas), 2002, 38(1), 36 - 42 {Dependence of peripheral blood lymphocyte subpopulations on causative microorganisms able to produce superantigens}; Verba V et al.; A retrospective study of 176 immunologically tested patients admitted to Kaunas Medical University Hospital during 1997-2000 was performed . All patients had positive bacteriological culture test result confirming an infectious etiology of the disease . Our results showed that majority of immunological parameters were dependent on such non-specific factors as intensity and localization of the inflammatory process, an overall functional status of the patient, and the number of the disease exacerbation episodes during the last year before admission . In contrast to this, the absolute number of CD4 lymphocytes, the relative amount of HLA-DR positive lymphocytes and the index of neutrophil latex phagocytosis were exceptionally dependent on the species of the causative microorganism, in particular on superantigen producing cocci . In this case, the HLA-DR positive lymphocyte amount and the neutrophil phagocytosis index were significantly higher . In addition, the CD4/CD8 lymphocyte ratio (the immunoregulatory index) was significantly lower in this group . As much as those findings are concordant with the signs of excessive immune activation, we conclude that they reflect a possible superantigenic action of the disease causing bacteria . Therefore, a need for immunomodulating therapy during the infections caused by species able to produce superantigens is confirmed. Pol Merkuriusz Lek, 2002 Sep, 13(75), 238 - 41 {The role of monocytes/macrophages and their cytokines in the development of immunosuppression after severe injury}; Jedynak M et al.; Despite new generations of antibiotics and great improvement in the fields of rescue and modern intensive care medicine, sepsis remains one of the most frequent causes of complications and death in severely injured patients . Shock, bacterial colonization, invasive methods of treatment and immune mechanisms are reported to be responsible for the increased susceptibility of patients to sepsis after trauma . Posttraumatic immune abnormalities consist of two mechanistic entities: inappropriately hyperactive inflammatory processes and profound depression of cell-mediated immune function . Monocytes/macrophages carry out the fundamental protective functions of ingesting and killing invading microorganisms . Macrophages play a central role in the immune response by presenting antigens to lymphocytes, modulating T cell functions and by secreting a large number of inflammatory mediators . Macrophage-derived cytokines play key roles in the amplification of both humoral and cell-mediated immune responses . Depression of macrophage function appears to have serious deleterious effects in critically injured patients and has been associated with increased mortality . Phagocytosis, oxidative burst activity and cytokine secretion are impaired in macrophages early after traumatic injury . However, a widely accepted clinical treatment for post-injury immunosuppression does not currently exist . As our understanding of the pathogenesis of injury-induced immunosuppression progresses, our treatment approaches will likely improve. Clin Cancer Res, 2002 Dec, 8(12), 3877 - 84 Interleukin 15 promotes antigen-independent in vitro expansion and long-term survival of antitumor cytotoxic T lymphocytes; Lu J et al.; The survival and expansion of effector cytotoxic T lymphocytes (CTLs) during an immunological response are critical for the successful elimination of life-threatening attacks by microorganisms, parasites, or malignant cells . Among the numerous factors that regulate the immune response, interleukin (IL)-2, and its close relative, IL-15 are known to function as growth and survival factors for antigen-experienced T cells . However, major differences appear to exist between these lymphokines in their capacity to act on various T-cell types such as CD4+ versus CD8+ or effector versus memory T lymphocytes . Although several studies have been done in the mouse system, less information is available regarding the function of these lymphokines in the human system . Here, we report that IL-15 or high concentrations of IL-2 induced antigen-independent expansion of effector CD8+ CTLs . Neither IL-2 nor IL-15 induced the proliferation of CD4+ T cells . In the absence of antigen, at least one of these lymphokines was required for the long-term survival of the cells in tissue culture . Most significantly, the effector cytolytic activity of CTLs expanded and maintained in IL-15 for up to 60 days remained stable, indicating that these cells do not differentiate into a memory functional phenotype . The expression of IL-15Ralpha, which was detected on CD8+ CTLs but not on CD4+ helper T cells, suggests that this receptor subunit somehow participates in the transduction of the mitogenic signals of IL-15 . The present findings have practical implications for the propagation of antigen-specific T-cell lines in vitro and could be useful for expansion of therapeutic T cells for adoptive transfer. Clin Infect Dis, 2002 Dec 15, 35(12), e141 - 4 Epub 2002 Nov 20. Vertebral osteomyelitis due to Bartonella henselae in adults: a report of 2 cases; Verdon R et al.; We describe 2 adult patients (1 of whom was infected with human immunodeficiency virus) with osteomyelitis due to Bartonella henselae . Diagnosis was established on the basis of direct identification of the microorganism in one case and seroconversion in the other . Both patients recovered completely within 3 months. J Environ Qual, 2002 Nov-Dec, 31(6), 2045 - 50 Volatilization and degradation of soil-applied dimethylselenide; Dungan RS et al.; Dimethylselenide (DMSe) is a highly volatile gas that is produced by indigenous microorganisms in seleniferous soils and sediments; however, little is known about the soil conditions that affect the persistence of DMSe and its transport to the atmosphere . In this study we investigated the effect of moisture content, temperature, and organic amendments on the degradation of soil-applied DMSe . The degradation of DMSe was entirely a result of biological mechanisms, but changes in temperature (20-40 degrees C) and soil moisture content (30-70% of the maximum water holding capacity) had little influence on the degradation rate . In contrast, amending soil with either 1% casein or gluten (by weight) had an inhibitory effect on the degradation of DMSe . After 18 d, 2.1 times more DMSe was present in the casein-amended soil and 2.6 times more DMSe was present in the gluten-amended soil . The transport of DMSe in packed soil columns was also investigated . Increasing the depth to soil surface was found to significantly decrease the amount of DMSe transported to the air . After 6 d, 57% of DMSe injected 10 cm below the soil surface was volatilized . At an injection depth of 20 cm the cumulative emissions were reduced by 38% and at 30 cm the cumulative emissions were reduced by 51% . In columns containing 1% casein or gluten in the top 5 cm of soil the cumulative loss of DMSe was about 9% higher than in unamended soil . Increasing our understanding of the soil conditions that influence the gaseous diffusion of DMSe should help in determining the feasibility of using Se volatilization as a remediation technique. Org Lett, 2002 Dec 12, 4(25), 4483 - 6 A new strategy toward the total synthesis of stachyflin, a potent anti-influenza A virus agent: concise route to the tetracyclic core structure; Nakatani M et al.; {reaction: see text} A new strategy directed toward the total synthesis of stachyflin, a potent and novel anti-influenza A virus agent isolated from a microorganism, has been presented through the enantioselective synthesis of the tetracyclic core structure . The synthetic method features a BF(3) x Et(2)O-induced domino epoxide-opening/rearrangement/cyclization reaction as the key step. Nucl Med Commun, 2002 Dec, 23(12), 1177 - 82 Detection of inflammatory lymph nodes in rabbits by 99mTc-HIG lymphoscintigraphy; Ergun EL et al.; Tc-Human immunoglobulin G ( Tc-HIG) is a well-known radiopharmaceutical for the evaluation of inflammatory lesions . Recently, it has been demonstrated as a new agent for the visualization of the lymphatic system by our group . Our aim was to investigate the feasibility of detection of inflammatory lymph nodes by Tc-HIG lymphoscintigraphy . Ten adult New Zealand rabbits were used as group A . In a baseline study, 37 MBq Tc-HIG (0.1 ml) was injected into both hind legs of the rabbits, and sequential posterior gamma imaging with the rabbits lying prone was performed at 5, 15, 30, 60, 90 and 120 min using a single-headed gamma camera (Toshiba GCA G01 E) . One week later, microorganisms ( ) were injected in a volume of 0.1 ml intradermally into the web space between the second and third toes in the bilateral hind legs of each rabbit in order to obtain inflammation in the popliteal lymph nodes . After 4 days, 37 MBq Tc-HIG (0.1 ml) was injected into the hind legs of the rabbits bilaterally, and sequential posterior gamma imaging was performed as described above (second study) . Another group of 10 adult New Zealand rabbits (group B) was injected with the same microorganisms in the right hind legs only . After 4 days, scintigraphic imaging was carried out in the same way as described above (third study) . Regions of interest were drawn over the injection sites and popliteal lymph nodes on each image for semiquantitative analysis . Count rates for each were calculated and a decay correction was applied . Time-activity curves were generated to show the percentage retention of radioactivity in each region . After the scintigraphic study, some of the group B rabbits were killed by intravenous injection of pentobarbitone (100-150 mg.kg, and both left and right lymph nodes were removed for microscopic examination . On the scintigrams, lymphatic channels and popliteal lymph nodes were visualized within 15 min . In the second study, bilateral popliteal lymph nodes were visualized more clearly than in the baseline study . The right popliteal lymph nodes of the rabbits were more clearly visualized in the third study . Semiquantitative analysis showed a higher percentage uptake of radioactivity in the right compared to the left popliteal lymph nodes in group B rabbits . Microscopic examination of the tissue sections demonstrated inflammation in the right lymph nodes of group B rabbits . In this preliminary study, it was found that Tc-HIG is a new promising agent for the demonstration and evaluation of inflammatory lymph nodes. J Chromatogr A, 2002 Nov 8, 976(1-2), 103 - 11 Investigating the effects of protein patterns on microorganism identification by high-performance liquid chromatography-mass spectrometry and protein database searches; Ho YP et al.; High-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS) has been employed for separation and detection of protein biomarkers from E . coli samples . LC-MS is suitable for microbial identification because it can couple on-line with sample clean-up devices and is readily amenable to automation . In this work, we have investigated the effects of sample preparation methods on the detection of bacterial proteins by LC-MS . Many factors effect the degree of variations in the protein patterns (i.e . number and masses of proteins) . For example, changing the polarity as well as pH of the extraction solvent may control the number of detected proteins . It is also noted that the protein patterns can vary even when the total ion chromatography plots seem to be the same under the same sample preparation conditions . Further, we have tested experimentally the influence of LC-MS-analyzed protein patterns (molecular masses between 2000 and 60,000) on microbial identification by protein database searches . This is in contrast to the current database search approach, where only the masses of smaller proteins (< or = 20,000) from direct matrix-assisted laser/desorption ionization MS are used . In spite of the variations in protein patterns, all the database search results show that the best matches come from the correct microorganism. Int J Occup Med Environ Health, 2002, 15(3), 303 - 8 Firefighting efforts may lead to massive fungal growth and exposure within one week . A case report; Rautiala SH et al.; A case study on extensive fungal growth that occurred in an apartment building after firefighting efforts is described in this paper . Exposure to airborne microorganisms (both viable and total) was investigated by filter sampling in three periods before and during remedial actions after the fire . Material samples were also analyzed . Extensive mold growth was observed on the building materials as soon as eight days after the fire . High concentrations of fungal spores, 10(7) cfu/g, were found when material samples were analyzed . Concentrations of airborne fungal spores (10(4) spores/m3) were also high and increased by two orders of magnitude during the demolition of moldy building materials and during the clean-up after the demolition . The proportions of airborne viable fungi in comparison with the total spore concentrations were 28-83% immediately after the fire, but they had decreased to <1% two months after the fire during the reconstruction phase . Paecilomyces was the main fungal genus in the indoor air before and during the demolition, while Penicillium dominated during the reconstruction . Paecilomyces was not detected in the outdoor air . Paecilomyces and Penicillium were also found in the material samples . The results show that fast and extensive mold growth in a building may take place also in subarctic climates, at least during summer . High concentrations of fungal spores are released to the air during the demolition of moldy building materials and the following clean-up . Therefore, personal protection is necessary during such work. Ocul Immunol Inflamm, 2002 Mar, 10(1), 27 - 39 Distribution, markers, and functions of retinal microglia; Chen L et al.; Retinal microglia originate from hemopoietic cells and invade the retina from the retinal margin and the optic disc, most likely via the blood vessels of the ciliary body and iris, and the retinal vasculature, respectively . The microglial precursors that appear in the retina prior to vascularization are major histocompatibility complex (MHC) class I- and II-positive and express the CD45 marker, but lack specific macrophage markers . They differentiate into ramified parenchymal microglia in the adult retina . A second category of microglial precursors, which do express specific macrophage markers, migrate into the retina along with vascular precursors . They appear around blood vessels in the adult retina and are similar to macrophages or cells of the mononuclear phagocyte series (MPS) . Microglia are distributed in the outer plexiform layer (OPL), outer nuclear layer (ONL), inner plexiform layer (IPL), ganglion cell layer (GCL), and nerve fiber layer (NFL) of the primate retina . The pattern of microglial distribution in the avascular retina of the quail indicates that blood vessels are not responsible for the final location of microglia in the retina . In the human retina, microglia express MHC class I, MHC class II, CD45, CD68, and S22 markers . In the rat and mouse retina, OX41, OX42, OX3, OX6, OX18, ED1, Mac-1, F4/80, 5D4 anti-keratan sulfate, and lectins are used to recognize microglia . Microglial cells play an important role in host defense against invading microorganisms, immunoregulation, and tissue repair . During neurodegeneration, activated microglial cells participate in the phagocytosis of debris and facilitate regenerative processes . In autoimmune disease, microglia have dual functions: initiating uveoretinitis, but also limiting subsequent inflammation . Retinal microglia may be associated with vitreoretinopathy, diabetic retinopathy, glaucoma, and age-related macular degeneration . The goal of this article was to review the present knowledge about retinal microglia and the function of retinal microglia in pathological conditions. Am J Infect Control, 2002 Dec, 30(8), S1 - 46 Guideline for Hand Hygiene in Health-Care Settings . Recommendations of the Healthcare Infection Control Practices Advisory Committee and the HIPAC/SHEA/APIC/IDSA Hand Hygiene Task Force; Boyce JM et al.; The Guideline for Hand Hygiene in Health-Care Settings provides health-care workers (HCWs) with a review of data regarding handwashing and hand antisepsis in health-care settings . In addition, it provides specific recommendations to promote improved hand-hygiene practices and reduce transmission of pathogenic microorganisms to patients and personnel in health-care settings . This report reviews studies published since the 1985 CDC guideline (Garner JS, Favero MS . CDC guideline for handwashing and hospital environmental control, 1985 . Infect Control 1986;7:231-43) and the 1995 APIC guideline (Larson EL, APIC Guidelines Committee . APIC guideline for handwashing and hand antisepsis in health care settings . Am J Infect Control 1995;23:251-69) were issued and provides an in-depth review of hand-hygiene practices of HCWs, levels of adherence of personnel to recommended handwashing practices, and factors adversely affecting adherence . New studies of the in vivo efficacy of alcohol-based hand rubs and the low incidence of dermatitis associated with their use are reviewed . Recent studies demonstrating the value of multidisciplinary hand-hygiene promotion programs and the potential role of alcohol-based hand rubs in improving hand-hygiene practices are summarized . Recommendations concerning related issues (e.g., the use of surgical hand antiseptics, hand lotions or creams, and wearing of artificial fingernails) are also included. Environ Microbiol, 2002 Nov, 4(11), 770 - 3 A fluorimetric method for the estimation of G+C mol% content in microorganisms by thermal denaturation temperature; Gonzalez JM et al.; G+C mol% content in microorganisms is one of the recommended characteristics for the standard description of bacterial species . In this study we present a novel fluorimetric method to estimate the G+C mol% content in microorganisms . Double-stranded DNA was specifically stained with SYBR Green I, and its thermal denaturalization was followed by measuring a decrease in fluorescence using a real-time PCR thermocycler . Unlike most previous determinations of G+C mol%, in this study only DNA from microorganisms with an available completely sequenced genome were used to prepare the calibration curves . Calibration curves showed a linear relationship between G+C mol% content and melting temperature and they were performed both in the absence and presence of 30% formamide . This protocol proves to be a rapid and inexpensive method to estimate DNA base ratios of novel microorganisms. Environ Microbiol, 2002 Nov, 4(11), 654 - 66 Cultivation of globally distributed soil bacteria from phylogenetic lineages previously only detected in cultivation-independent surveys; Sait M et al.; The culturability of microorganisms in a 10 cm core of an Australian pasture soil was investigated using a minimal agar medium with xylan as the growth substrate . Culturability decreased with increasing depth, from a maximum of 19% of the total microscopically countable cells in the 0-2 cm section to 2.4% in the 8-10 cm section . Seventy-one isolates from the core were identified by comparative 16S rRNA gene sequence analysis . Many of these isolates belong to groups of globally distributed soil bacteria, including well-characterized families of the classes Alphaproteobacteria and Betaproteobacteria, and of the subclass Actinobacteridae . Other isolates belong to groups with few or no cultivated representatives: 10 isolates in two subdivisions of the phylum Acidobacteria, five isolates in a new order and nine isolates in a new family of the class Alphaproteobacteria, two isolates in a new order of the class Gammaproteobacteria, three isolates in two new families of the subclass Actinobacteridae, and two isolates in the subclass Rubrobacteridae . These new isolates represent the first laboratory cultures able to be assigned to some of these groups and greatly increase the number of cultivated strains known for others . This demonstrates that a minimal change in cultivation strategy (using a polymeric growth substrate and longer incubation times) can result in the isolation of globally distributed but previously uncultured phylogenetically novel soil bacteria. J Microbiol Methods, 2003 Feb, 52(2), 267 - 71 Development of a fluorescence in situ hybridization method for cheese using a 16S rRNA probe; Ercolini D et al.; A 16S rRNA fluorescence in situ hybridization (FISH) method for cheese was developed to allow detection in situ of microorganisms within the dairy matrix . An embedding procedure using a plastic resin was applied to Stilton cheese, providing intact embedded cheese sections withstanding the hybridization reaction . The use of a fluorescein-labelled 16S rRNA Domain Bacteria probe allowed observation of large colonies of microbial cells homogeneously distributed in the cheese matrix . FISH experiments performed on cheese suspensions provided images of the different microbial morphotypes occurring . The technique has great potential to study the spatial distribution of microbial populations in situ in foods, especially where the matrix is too fragile to allow manipulation of cryosections . Turk J Pediatr, 2002 Oct-Dec, 44(4), 283 - 8 Frequency of Mycoplasma pneumoniae among atypical pneumonia of childhood; Oguz F et al.; We aimed to investigate the frequency of Mycoplasma pneumoniae among atypical pneumonia of childhood that is acquired from the community and to determine a practical approach to the diagnosis of these patients . In this prospective study, 55 patients (31 male and 24 female) with atypical pneumonia were investigated with conventional laboratory and radiological methods as well as culture and polymerase chain reaction (PCR) on throat swab . In addition, serum of the patients was tested for M . pneumoniae specific IgM . The patients were reevaluated clinically at 3-5 days and 3-4 weeks and serologically at 3-4 weeks . The data on patients with M . pneumoniae pneumonia were compared with the other patients with atypical pneumonia and controls . All patients were treated with macrolide antibiotics . The mean age of the patients was 7.8+/-2.9 years . The frequency of M . pneumoniae by this method was 34.5% . Neither clinical, laboratory, or epidemiological data nor response to macrolide antibiotics was useful in detecting the etiology of atypical pneumonia . Diagnostic sensitivity and specificity of IgM+IgG antibodies plus PCR on throat swab were estimated as 100% . M . pneumoniae was an important microorganism in the etiology of atypical pneumonia of childhood in our community . In order to prevent loss of time with beta-lactamase antibiotics, which are usually started in severe pneumonia, serologic tests and PCR must be done during the initial evaluation of the patient for the reliable diagnosis of M . pneumoniae, which will increase the chance of early and appropriate therapy. Anal Bioanal Chem, 2002 Nov, 374(6), 1101 - 4 Epub 2002 Oct 19. Empirical method for prediction of the coordination environment of Eu(III) by time-resolved laser-induced fluorescence spectroscopy; Ozaki T et al.; The number of water molecules in the inner-sphere (N(H2O)) was determined for Eu(III) and the strength of ligand field (R(E/M)) was evaluated for a variety of coordination environments from the luminescence lifetime and the relative intensity at 615 nm and at 592 nm, by time-resolved laser-induced fluorescence spectroscopy . When R(E/M) and deltaN(H2O) for Eu(III) with a known coordination environment were plotted clear regularity was apparent between the location of the R(E/M)-deltaN(H2O) plot and the coordination environment of Eu(III) . Here, deltaN(H2O) was calculated by use of the equation, deltaN(H2O)=9-N(H2O) . Unknown coordination environments of Eu(III) can, in turn, be characterized, including both the inner- and the outer-sphere, simply by plotting R(E/M) and deltaN(H2O) for Eu(III) on the diagram . This empirical method is effective for prediction of the coordination environment of hydrated and complexed Eu(III) in solutions and that of the adsorbed Eu(III) on ion-exchange resins and by microorganisms. J Biol Chem, 2003 Feb 7, 278(6), 3552 - 61 Epub 2002 Nov 26. Prophenoloxidase-activating proteinase-2 from hemolymph of Manduca sexta . A bacteria-inducible serine proteinase containing two clip domains; Jiang H et al.; Proteolytic activation of prophenoloxidase in insects is a component of the host defense system against invading pathogens and parasites . We have purified from hemolymph of the tobacco hornworm, Manduca sexta, a new serine proteinase that cleaves prophenoloxidase . This enzyme, designated prophenoloxidase-activating proteinase-2 (PAP-2), differs from another PAP, previously isolated from integuments of the same insect (PAP-1) . PAP-2 contains two clip domains at its amino terminus and a catalytic domain at its carboxyl terminus, whereas PAP-1 has only one clip domain . Purified PAP-2 cleaved prophenoloxidase at Arg(51) but yielded a product that has little phenoloxidase activity . However, in the presence of two serine proteinase homologs, active phenoloxidase was generated at a much higher level, and it formed covalently linked, high molecular weight oligomers . The serine proteinase homologs associate with a bacteria-binding lectin in M . sexta hemolymph, indicating that they may be important for ensuring that the activation of prophenoloxidase occurs only in the vicinity of invading microorganisms . PAP-2 mRNA was not detected in naive larval fat body or hemocytes, but it became abundant in these tissues after the insects were injected with bacteria. Front Biosci, 2003 Jan 01, 8, e23 - 35 Coxsackievirus experimental heart diseases; Gauntt C et al.; Many microorganisms, particularly viruses, can cause myocarditis, an inflammatory disease of the heart . The frequency of and major factors that contribute to this disease, including a pronounced gender (male) bias, age and genetic background parameters are discussed, along with signs and symptoms of disease in infants to adults . Individuals with acute disease generally recover without sequelae; the chronic form can develop into idiopathic dilated cardiomyopathy and death can follow . Among viruses most frequently associated with cases in the U.S., the coxsackieviruses group B (CVB) are major etiologic agents . The association between the CVB and disease is based on detection of viral RNA in heart biopsy specimens by polymerase chain reaction assays . Excellent CVB-, particularly coxsackievirus B3 (CVB3)-, mouse models of the disease have identified mechanisms of induction and establishment of chronic myocarditis . CVB3-murine models share many biologic parameters of the acute and chronic diseases in humans, and show that cardiopathologic alterations result from virus-induced and immunologic reactions in heart tissues . Several immune responses to a CVB3 infection that become cardiopathogenic, instead of protective, are discussed in an attempt to explain why immunosuppressive treatments are not effective . Bed rest and supportive therapy are the current treatment for patients with myocarditis. Contemp Top Lab Anim Sci, 1998 Mar, 37(2), 50 - 51 Effectiveness of Rack Sanitation Procedures for Elimination of Bacteria from Automatic Watering Manifolds; Costello T et al.; An important responsibility of animal care programs is to protect research animals from exposure to potentially pathogenic microorganisms . To validate the need for steam sterilization of rodent automatic watering racks, we evaluate the post-sanitation microbial contamination of experimentally inoculated racks and of racks that had been used to house conventional mice . We tested three sanitation protocols: rack-washer sanitation without manifold flush, sanitation that included manifold flush, and sanitation that included manifold flush followed by autoclaving . Rack sanitation, with or without manifold flush, did not reliably eliminate microbial flora from the lixits or manifold drainage water . A total of 43% of all non-autoclaved racks were positive for bacterial contamination after sanitation, and racks that had been used for conventional animal housing were more frequently positive than were experimentally inoculated racks (79% vs 18%) . These data indicate that steam sterilization is necessary for eliminating bacteria from automatic watering systems . These observations are particularly important in light of increasing numbers of immune-impaired rodents that may be inadvertently and unnecessarily exposed to microbial pathogens via the automatic watering system. J Clin Microbiol, 2002 Dec, 40(12), 4771 - 5 Are there naturally occurring pleomorphic bacteria in the blood of healthy humans? McLaughlin RW, Vali H, Lau PC, Palfree RG, De Ciccio A, Sirois M, Ahmad D, Villemur R, Desrosiers M, Chan EC. Dark-field microscopy of blood from healthy individuals revealed the existence of pleomorphic microorganisms . These bacteria exhibited limited growth and susceptibility to antibiotics and could be detected by fluorescent in situ hybridization and flow cytometry . They were further characterized by analysis of their 16S rRNA and gyrB genes. Emerg Infect Dis, 2002 Nov, 8(11), 1342 - 6 Genomewide pattern of synonymous nucleotide substitution in two complete genomes of Mycobacterium tuberculosis; Hughes AL et al.; Comparison of the pattern of synonymous nucleotide substitution between two complete genomes of Mycobacterium tuberculosis at 3298 putatively orthologous loci showed a mean percent difference per synonymous site of 0.000328 0.000022 . Although 80.5% of loci showed no synonymous or nonsynonymous nucleotide differences, the level of polymorphism observed at other loci was greater than suggested by previous studies of a small number of loci . This level of nucleotide difference leads to the conservative estimate that the common ancestor of these two genotypes occurred approximately 35000 ago, which is twice as high as some recent estimates of the time of origin of this species . Our results suggest that a large number of loci should be examined for an accurate assessment of the level of nucleotide diversity in natural populations of pathogenic microorganisms. Clin Exp Immunol, 2002 Dec, 130(3), 449 - 58 The scavenger receptor, cysteine-rich domain-containing molecule gp-340 is differentially regulated in epithelial cell lines by phorbol ester; Kang W et al.; Gp-340 is a glycoprotein belonging to the scavenger receptor cysteine rich (SRCR) group B family . It binds to host immune components such as lung surfactant protein D (SP-D) . Recent studies found that gp-340 interacts directly with pathogenic microorganisms and induces their aggregation, suggesting its involvement in innate immunity . In order to investigate further its potential immune functions in the appropriate cell lines, the expression of gp-340 in four conventional immune cell lines (U937, HL60, Jurkat, Raji), and two innate immune-related epithelial cell lines (A549 derived from lung and AGS from stomach), was examined by RT-PCR and immunohistochemistry . The resting immune cell lines showed weak or no gp-340 mRNA expression; while the two epithelial cell lines expressed gp-340 at much higher level, which was differentially regulated by phorbol myristate acetate (PMA) treatment . In the A549 cells, gp-340 was up-regulated along with the PMA-induced proinflammatory expression of both IL-6 and IL-8 . In AGS cells, PMA down-regulation of gp-340 was seen in parallel with an up-regulation of the two mature gastric epithelial specific proteins TFF1 (trefoil factor 1) and TFF2, which are implicated as markers of terminal differentiation . Analysis of the distribution of gp-340, together with the TFFs and SP-D in normal lung and gastric mucosa, supported further our in vitro data . We conclude that the differential regulation of gp-340 in the two epithelial cell lines by PMA indicates that gp-340 s involvement in mucosal defence and growth of epithelial cells may vary at different body locations and during different stages of epithelial differentiation. Jpn Heart J, 2002 Sep, 43(5), 475 - 85 Pacemaker related endocarditis: analysis of seven cases; Erdinler I et al.; Vegetative electrode infection following permanent pacemaker implantation is a rare and serious complication . Among 1920 patients who underwent permanent pacemaker implantation in our institute between 1980 and 2000, 7 patients aged 65 to 78 years were diagnosed to have pacemaker related endocarditis . In this study, the clinical course and management strategies for these patients are reviewed . The most frequently encountered factors contributing to development of pacemaker infection were local complications such as postoperative hematoma and inflammation, and recurrent surgical interventions on the pacemaker system . In blood cultures S . aureus was the most common causative microorganism . Echocardiography could be performed in 5 patients . Three patients were referred to open-heart surgery for total removal of the pacemaker system, and one patient had his pacemaker system removed percutaneously . The remaining 3 patients did not agree to either surgical or percutaneous removal . These patients have been under antibiotic therapy for approximately 3 years and they still do not have any signs of a serious infection . Consequently, in patients with permanent pacemakers, infective endocarditis should be considered in the presence of fever and local symptoms . Blood cultures should be obtained and echocardiography should be performed . Complete removal of the pacemaker system with intensive antibiotic treatment is necessary for complete eradication of the infection . However, if percutaneous or surgical removal of the electrodes cannot be done because of high perioperative risk or the patient does not agree to undergo either method, medical treatment with long term antibiotic use may be considered as an alternative. Appl Environ Microbiol, 2002 Dec, 68(12), 6403 - 4 The environmental pathogen Mycobacterium ulcerans grows in amphibian cells at low temperatures; Drancourt M et al.; Mycobacterium ulcerans, the etiological agent of Buruli ulcers, is an environmental pathogen . We cultivated it in an amphibian (XTC-2) cell line that grows at 28 degrees C . By counting of Ziehl-Neelsen-stained mycobacteria and by quantitative PCR analysis, we found that M . ulcerans multiplies rapidly in association with XTC-2 cells . Transmission electron microscopy demonstrated the presence of intracellular M . ulcerans microorganisms . These data suggest an intracellular environmental niche, and we propose use of XTC-2 cells for isolation of M . ulcerans from environmental sources. Appl Environ Microbiol, 2002 Dec, 68(12), 6392 - 8 Bacterial group II introns in a deep-sea hydrothermal vent environment; Podar M et al.; Group II introns are catalytic RNAs and mobile retrotransposable elements known to be present in the genomes of some nonmarine bacteria and eukaryotic organelles . Here we report the discovery of group II introns in a bacterial mat sample collected from a deep-sea hydrothermal vent near 9 degrees N on the East Pacific Rise . One of the introns was shown to self-splice in vitro . This is the first example of marine bacterial introns from molecular population structure studies of microorganisms that live in the proximity of hydrothermal vents . These types of mobile genetic elements may prove useful in improving our understanding of bacterial genome evolution and may serve as valuable markers in comparative studies of bacterial communities. Appl Environ Microbiol, 2002 Dec, 68(12), 6121 - 8 Molecular analysis of the nitrate-reducing community from unplanted and maize-planted soils; Philippot L et al.; Microorganisms that use nitrate as an alternative terminal electron acceptor play an important role in the global nitrogen cycle . The diversity of the nitrate-reducing community in soil and the influence of the maize roots on the structure of this community were studied . The narG gene encoding the membrane bound nitrate reductase was selected as a functional marker for the nitrate-reducing community . The use of narG is of special interest because the phylogeny of the narG gene closely reflects the 16S ribosomal DNA phylogeny . Therefore, targeting the narG gene provided for the first time a unique insight into the taxonomic composition of the nitrate-reducing community in planted and unplanted soils . The PCR-amplified narG fragments were cloned and analyzed by restriction fragment length polymorphism (RFLP) . In all, 60 RFLP types represented by two or more clones were identified in addition to the 58 RFLP types represented by only one clone . At least one clone belonging to each RFLP type was then sequenced . Several of the obtained sequences were not related to the narG genes from cultivated bacteria, suggesting the existence of unidentified nitrate-reducing bacteria in the studied soil . However, environmental sequences were also related to NarG from many bacterial divisions, i.e., Actinobacteria and alpha, beta, and gamma proteobacteria . The presence of the plant roots resulted in a shift in the structure of the nitrate-reducing community between the unplanted and planted soils . Sequencing of RFLP types dominant in the rhizosphere or present only in the rhizosphere revealed that they are related to NarG from the Actinobacteria in an astonishingly high proportion. Appl Environ Microbiol, 2002 Dec, 68(12), 6106 - 13 Linking of microorganisms to phenanthrene metabolism in soil by analysis of (13)C-labeled cell lipids; Johnsen AR et al.; Phenanthrene-metabolizing soil microbial communities were characterized by examining mineralization of {(14)C}phenanthrene, by most-probable-number (MPN) counting, by 16S-23S spacer DNA analysis of the numerically dominant, culturable phenanthrene-degrading isolates, and by examining incorporation of {(13)C}phenanthrene-derived carbon into sterols and polar lipid fatty acids (PLFAs) . An unpolluted agricultural soil, a roadside soil diffusely polluted with polycyclic aromatic hydrocarbons (PAHs), and two highly PAH-polluted soils from industrial sites were analyzed . Microbial phenanthrene degraders were not detected by MPN counting in the agricultural soil and the roadside soil . In the industrial soils, phenanthrene degraders constituted 0.04 and 3.6% of the total number of CFU . 16S-23S spacer DNA analysis followed by partial 16S DNA sequencing of representative isolates from one of the industrial soils showed that one-half of the isolates belonged to the genus Sphingomonas and the other half were closely related to an unclassified beta-proteobacterium . The (13)C-PLFA profiles of the two industrial soils were relatively similar and resembled the profiles of phenanthrene-degrading Sphingomonas reference strains and unclassified beta-proteobacterium isolates but did not match the profiles of Pseudomonas, Mycobacterium, or Nocardia reference strains . The (13)C-PLFA profiles of phenanthrene degraders in the agricultural soil and the roadside soil were different from each other and different from the profiles of the highly polluted industrial soils . Only in the roadside soil were 10me/12me18:0 PLFAs enriched in (13)C, suggesting that actinomycetes metabolized phenanthrene in this soil . The (13)C-PLFA profiles of the unpolluted agricultural soil did not resemble the profiles of any of the reference strains . In all of the soils investigated, no excess (13)C was recovered in the 18:2omega6,9 PLFA, suggesting that fungi did not contribute significantly to assimilation of {(13)C}phenanthrene. Med Hypotheses, 2003 Jan, 60(1), 69 - 83 Are lipid peroxidation processes induced by changes in the cell wall structure and how are these processes connected with diseases? Spiteller G. Apparently nature uses the unique sensitivity of polyunsaturated fatty acids (PUFAs) versus oxygen to generate chemical signals if the surface of a cell is influenced by an outside or inside event; for instance the attack of microorganisms, proliferation, aging or by treatment of isolated cells with surfactants . It seems that mammalian and plant cells respond equally to such changes in their structures by transformation of polyunsaturated fatty acids localized in the phospholipid layer of the cell wall to lipidhydroperoxides (LOOHs) . These lipid peroxidation (LPO) processes involve all PUFAs, not only arachidonic acid.Slight physiological changes of the cell wall for instance by proliferation seem to activate enzymes, e.g., phospholipases and lipoxygenases (LOX) . When an outside impact (for instance by attack of microorganisms) exceeds a certain level LOX commit suicide and liberate iron ions . These start a nonenzymatic LPO . Enzymatic and nonenzymatic LPO distinguish fundamentally which has not been recognized in the past . In the enzymatic LPO processes peroxyl radicals generated as intermediates cannot leave the enzyme complex . In contrast in a nonenzymatic LPO process peroxyl radicals are not trapped . They attack nearly any kind of biological molecules, for instance proteins . Thus only the amount of an outside impact decides if proliferation, apoptosis, or necrosis is started.Some evidence indicates that cancer might be the consequence of a low response of cells to induce apoptotic LPO processes . In contrast to high level of LPO processes induces diseases combined with inflammation, for instance rheumatic arthritis . After consumption of food rich in linoleic acid its LPO products become increased in low density lipoprotein (LDL) . This LDL is able to enter endothelial cells and damage cells from inside, long before an inflammatory response is detectable. Gene Expr, 2002, 10(5-6), 213 - 30 A DNA microarray-based approach to elucidate the effects of the immunosuppressant SR31747A on gene expression in Saccharomyces cerevisiae; Cinato E et al.; SR31747A is an immunosuppressive agent that arrests cell proliferation in the yeast Saccharomyces cerevisiae . In this microorganism, SR31747A was shown to inhibit the ERG2 gene product, namely the delta8-delta7 sterol isomerase, involved in the ergosterol biosynthesis pathway . Although previous genetic experiments pointed to this enzyme as the target for SR31747A in yeast, the existence of other potential targets could not be ruled out . To enlighten this issue, we undertook a DNA microarray-based approach in which the expression profile of SR31747A-treated wild-type cells defining the "drug signature" was compared with the "mutant signature," the expression profile of the corresponding ERG2-deleted strain . We observed that treatment of ERG2-positive cells with SR31747A resulted in the modulation of mRNA levels of numerous genes . Among them, 121 werealso affected in untreated ERG2-disrupted cells compared with wild-type cells . By contrast, drug exposure did not induce any significant transcriptional change in the ERG2 null mutant . These results were consistent with SR31747A being an inhibitor of the sterol isomerase and demonstrated the absence of any additional SR31747A target . The detailed analysis of the observed 121 modulated genes provides new insights into the cellular response to ergosterol deprivation induced by SR31747A through inhibition of the ERG2 gene product. Astrobiology, 2002 Spring, 2(1), 77 - 82 Aspartic acid racemization and age-depth relationships for organic carbon in Siberian permafrost; Brinton KL et al.; We have analyzed the degree of racemization of aspartic acid in permafrost samples from Northern Siberia, an area from which microorganisms of apparent ages up to a few million years have previously been isolated and cultured . We find that the extent of aspartic acid racemization in permafrost cores increases very slowly up to an age of approximately 25,000 years (around 5 m in depth) . The apparent temperature of racemization over the age range of 0-25,000 years, determined using measured aspartic acid racemization rate constants, is -19 degrees C . This apparent racemization temperature is significantly lower than the measured environmental temperature (-11 to -13 degrees C) and suggests active recycling of D-aspartic acid in Siberian permafrost up to an age of around 25,000 years . This indicates that permafrost organisms are capable of repairing some molecular damage incurred while in a "dormant" state over geologic time. Astrobiology, 2002 Spring, 2(1), 59 - 76 Planetary resources and astroecology . Planetary microcosm models of asteroid and meteorite interiors: electrolyte solutions and microbial growth--implications for space populations and panspermia; Mautner MN; Planetary microcosms were constructed using extracts from meteorites that simulate solutions in the pores of carbonaceous chondrites . The microcosms were found to support the growth of complex algal and microbial populations . Such astroecology experiments demonstrate how a diverse ecosystem could exist in fluids within asteroids, and in meteorites that land on aqueous planets . The microcosm solutions were obtained by extracting nutrient electrolytes under natural conditions from powders of the Allende (CV3) and Murchison (CM2) meteorites at low (0.02 g/ml) and high (10.0 g/ml) solid/solution ratios . The latter solutions contain > 3 mol/L electrolytes and about 10 g/L organics, that simulate natural fluids in asteroids during aqueous alteration and in the pores of meteorites, which can help prebiotic synthesis and the survival of early microorganisms . These solutions and wet solids were in fact found to support complex self-sustaining microbial communities with populations of 4 x 10(5) algae and 6 x 10(6) bacteria and fungi for long periods (> 8 months) . The results show that planetary microcosms based on meteorites can: assay the fertilities of planetary materials; identify space bioresources; target astrobiology exploration; and model past and future space-based ecosystems . The results show that bioresources in the carbonaceous asteroids can sustain a biomass of 10(18) kg, comprising 10(32) microorganisms and a human population of 10(14) . The results also suggest that protoplanetary nebulae can support and disperse microorganisms and can be therefore effective environments for natural and directed panspermia. Astrobiology, 2002 Spring, 2(1), 43 - 57 Fluid inclusion studies of chemosynthetic carbonates: strategy for seeking life on Mars; Parnell J et al.; Fluid inclusions in minerals hold the potential to provide important data on the chemistry of the ambient fluids during mineral precipitation . Especially interesting to astrobiologists are inclusions in low-temperature minerals that may have been precipitated in the presence of microorganisms . We demonstrate that it is possible to obtain data from inclusions in chemosynthetic carbonates that precipitated by the oxidation of organic carbon around methane-bearing seepages . Chemosynthetic carbonates have been identified as a target rock for astrobiological exploration . Other surficial rock types identified as targets for astrobiological exploration include hydrothermal deposits, speleothems, stromatolites, tufas, and evaporites, each of which can contain fluid inclusions . Fracture systems below impact craters would also contain precipitates of minerals with fluid inclusions . As fluid inclusions are sealed microchambers, they preserve fluids in regions where water is now absent, such as regions of the martian surface . Although most inclusions are < 5 microns, the possibility to obtain data from the fluids, including biosignatures and physical remains of life, underscores the advantages of technological advances in the study of fluid inclusions . The crushing of bulk samples could release inclusion waters for analysis, which could be undertaken in situ on Mars. Zh Mikrobiol Epidemiol Immunobiol, 2002 Jul-Aug, (4), 63 - 72 {Integrative processes in modern epidemiology}; Litvin VIu et al.; The integration of achievements in population ecology and the molecular genetics of pathogenic microorganisms into nowadays epidemiology of infectious diseases and the formation of its corresponding trends are discussed . The decisive role of highly complicated key problems of general epidemiology, such as the regularities of the autochthonous existence of pathogenic bacteria in the environment, the mechanisms of the pathogenicity regulation of infective agents and the formation of their epidemic variants, the mechanisms of the prolonged reservation of pathogenic bacteria at interepidemic (interepizootic) periods, the mechanisms of transformation of active (vegetative) and dormant forms in bacterial populations at different stages of epidemic and epizootic cycles, is demonstrated . The detection and analysis of such processes, and especially their latent mechanisms, become possible with the use of new concepts and methodological approaches, highly productive for the progress of epidemiology. Mikrobiologiia, 2002 Sep-Oct, 71(5), 690 - 6 {The classification and the monitoring of the state of mouth riverine and lacustrine ecosystems in lake Baikal based on the composition of local microbiocenoses and their activity}; Maksimov VV et al.; The paper presents the results of the long-term investigation of microbial communities in the technogenically vulnerable mouth riverine and lacustrine ecosystems of Lake Baikal . The structural and functional parameters of the microbial communities were analyzed from the standpoint of developing destructive processes . The analysis showed that the total number of microorganisms (TNM), the number of saprophytic bacteria (NSB), and bacterial production (BP) were greater in the river-mouth water than in the near-mouth lake water . In the offshore direction, TNM and NSB decreased by a factor of 1.5 to 2, and BP decreased by a factor of 4 to 7 . Based on TNM, NSB, and BP data, we classified the Lake Baikal rivers with respect to the degree of the impact of human activities on them . The degrading capability of the riverine microbial communities was found to be such that they degrade daily from tenths of a percent to 3.5% of the total amount of organic compounds polluted the river waters. Eur Respir J, 2002 Nov, 20(5), 1351 - 3 Extrinsic allergic alveolitis induced by the yeast Debaryomyces hansenii; Yamamoto Y et al.; A 65-yr-old female developed cough, fever and dyspnoea following repeated exposure to a home ultrasonic humidifier . High-resolution computed tomography showed ground-glass opacity in both lung fields . Arterial blood gas analysis gave an oxygen tension of 8.38 kPa (63 Torr) . Pulmonary function testing revealed restrictive ventilatory impairment with a reduction in the diffusing capacity . The diagnosis of extrinsic allergic alveolitis (EAA) was confirmed by radiographic findings, pathological evidence of alveolitis and reproductive development by a provocation test to the humidifier water . The yeast Debaryomyces Hansenii was the only microorganism cultured from the water of the humidifier . The double diffusion precipitating test and lymphocyte proliferative response was positive for an extract of D . Hansenii, providing evidence to incriminate this fungus . This is the first described case of EAA caused by D . Hansenii. Astrobiology, 2001 Spring, 1(1), 89 - 109 The physics, biology, and environmental ethics of making mars habitable; McKay CP et al.; The considerable evidence that Mars once had a wetter, more clement, environment motivates the search for past or present life on that planet . This evidence also suggests the possibility of restoring habitable conditions on Mars . While the total amounts of the key molecules--carbon dioxide, water, and nitrogen--needed for creating a biosphere on Mars are unknown, estimates suggest that there may be enough in the subsurface . Super greenhouse gases, in particular, perfluorocarbons, are currently the most effective and practical way to warm Mars and thicken its atmosphere so that liquid water is stable on the surface . This process could take approximately 100 years . If enough carbon dioxide is frozen in the South Polar Cap and absorbed in the regolith, the resulting thick and warm carbon dioxide atmosphere could support many types of microorganisms, plants, and invertebrates . If a planet-wide martian biosphere converted carbon dioxide into oxygen with an average efficiency equal to that for Earth's biosphere, it would take > 100,000 years to create Earth-like oxygen levels . Ethical issues associated with bringing life to Mars center on the possibility of indigenous martian life and the relative value of a planet with or without a global biosphere. Astrobiology, 2001 Winter, 1(4), 447 - 65 Mineralogical biosignatures and the search for life on Mars; Banfield JF et al.; If life ever existed, or still exists, on Mars, its record is likely to be found in minerals formed by, or in association with, microorganisms . An important concept regarding interpretation of the mineralogical record for evidence of life is that, broadly defined, life perturbs disequilibria that arise due to kinetic barriers and can impart unexpected structure to an abiotic system . Many features of minerals and mineral assemblages may serve as biosignatures even if life does not have a familiar terrestrial chemical basis . Biological impacts on minerals and mineral assemblages may be direct or indirect . Crystalline or amorphous biominerals, an important category of mineralogical biosignatures, precipitate under direct cellular control as part of the life cycle of the organism (shells, tests, phytoliths) or indirectly when cell surface layers provide sites for heterogeneous nucleation . Biominerals also form indirectly as by-products of metabolism due to changing mineral solubility . Mineralogical biosignatures include distinctive mineral surface structures or chemistry that arise when dissolution and/or crystal growth kinetics are influenced by metabolic by-products . Mineral assemblages themselves may be diagnostic of the prior activity of organisms where barriers to precipitation or dissolution of specific phases have been overcome . Critical to resolving the question of whether life exists, or existed, on Mars is knowing how to distinguish biologically induced structure and organization patterns from inorganic phenomena and inorganic self-organization . This task assumes special significance when it is acknowledged that the majority of, and perhaps the only, material to be returned from Mars will be mineralogical. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 557 - 64 Mechanisms of natural soil suppressiveness to soilborne diseases; Mazzola M; Suppressive soils are characterized by a very low level of disease development even though a virulent pathogen and susceptible host are present . Biotic and abiotic elements of the soil environment contribute to suppressiveness, however most defined systems have identified biological elements as primary factors in disease suppression . Many soils possess similarities with regard to microorganisms involved in disease suppression, while other attributes are unique to specific pathogen-suppressive soil systems . The organisms operative in pathogen suppression do so via diverse mechanisms including competition for nutrients, antibiosis and induction of host resistance . Non-pathogenic Fusarium spp . and fluorescent Pseudomonas spp . play a critical role in naturally occurring soils that are suppressive to Fusarium wilt . Suppression of take-all of wheat, caused by Gaeumannomyces graminis var . tritici, is induced in soil after continuous wheat monoculture and is attributed, in part, to selection of fluorescent pseudomonads with capacity to produce the antibiotic 2,4-diacetylphloroglucinol . Cultivation of orchard soils with specific wheat varieties induces suppressiveness to Rhizoctonia root rot of apple caused by Rhizoctonia solani AG 5 . Wheat cultivars that stimulate disease suppression enhance populations of specific fluorescent pseudomonad genotypes with antagonistic activity toward this pathogen . Methods that transform resident microbial communities in a manner which induces natural soil suppressiveness have potential as components of environmentally sustainable systems for management of soilborne plant pathogens. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 549 - 56 Significance of lytic enzymes from Trichoderma spp . in the biocontrol of fungal plant pathogens; Viterbo A et al.; The use of specific mycolytic soil microorganisms to control plant pathogens is an ecological approach to overcome the problems caused by standard chemical methods of plant protection . The ability to produce lytic enzymes is a widely distributed property of rhizosphere-competent fungi and bacteria . Due to the higher activity of Trichoderma spp . lytic enzymes as compared to the same class of enzymes from other microorganisms and plants, effort is being aimed at improving biocontrol agents and plants by introducing Trichoderma genes via genetic manipulations . An overview is presented of the data currently available on lytic enzymes from the mycoparasitic fungus Trichoderma. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 509 - 20 Effects of above-ground plant species composition and diversity on the diversity of soil-borne microorganisms; Kowalchuk GA et al.; A coupling of above-ground plant diversity and below-ground microbial diversity has been implied in studies dedicated to assessing the role of macrophyte diversity on the stability, resilience, and functioning of ecosystems . Indeed, above-ground plant communities have long been assumed to drive below-ground microbial diversity, but to date very little is known as to how plant species composition and diversity influence the community composition of micro-organisms in the soil . We examined this relationship in fields subjected to different above-ground biodiversity treatments and in field experiments designed to examine the influence of plant species on soil-borne microbial communities . Culture-independent strategies were applied to examine the role of wild or native plant species composition on bacterial diversity and community structure in bulk soil and in the rhizosphere . In comparing the influence of Cynoglossum officinale (hound's tongue) and Cirsium vulgare (spear thistle) on soil-borne bacterial communities, detectable differences in microbial community structure were confined to the rhizosphere . The colonisation of the rhizosphere of both plants was highly reproducible, and maintained throughout the growing season . In a separate experiment, effects of plant diversity on bacterial community profiles were also only observed for the rhizosphere . Rhizosphere soil from experimental plots with lower macrophyte diversity showed lower diversity, and bacterial diversity was generally lower in the rhizosphere than in bulk soil . These results demonstrate that the level of coupling between above-ground macrophyte communities and below-ground microbial communities is related to the tightness of the interactions involved . Although plant species composition and community structure appear to have little discernible effect on microbial communities inhabiting bulk soil, clear and reproducible changes in microbial community structure and diversity are observed in the rhizosphere. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 357 - 64 The consequences of volatile organic compound mediated bacterial and fungal interactions; Wheatley RE; Microbial interactions via infochemicals are fundamental to the development of spatial distribution and activity variations in ecosystems . Microorganisms produce a wide range of infochemicals, frequently secondary metabolites, most of which are soluble and many volatile . Volatile organic compounds (VOCs) have been identified in soil atmospheres and related to community structure and function . VOC profiles produced by microorganisms are consistent, relating to cultural conditions, environment and inputs, and so to population and function dynamics . VOC-mediated interactions can result in functional responses by the organisms involved that result in selective advantage to some community members . Positive, negative or neutral interactions can occur between a very wide range of soil bacteria and fungi . These effects include both stimulation and inhibition of growth, by 40 and 60%, respectively, and enzyme production . These effects are usually transient, e.g . removal of an antagonist is followed by complete recovery . Up- and down-regulation of gene expression, by mRNA and protein profiling has been demonstrated . VOCs have played an important role during the evolution of microorganisms in the context of their communities. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 343 - 51 Mycorrhizosphere interactions to improve plant fitness and soil quality; Barea JM et al.; Arbuscular mycoruhizal fungi are key components of soil microbiota and obviously interact with other microorganisms in the rhizosphere, i.e . the zone of influence of plant roots on microbial populations and other soil constituents . Mycorrhiza formation changes several aspects of plant physiology and some nutritional and physical properties of the rhizospheric soil . These effects modify the colonization patterns of the root or mycovrhizas (mycorrhizosphere) by soil microorganisms . The rhizosphere of mycorrhizal plants, in practice a mycorrhizosphere, harbors a great array of microbial activities responsible for several key ecosystem processes . This paper summarizes the main conceptual principles and accepted statements on the microbial interactions between mycorrhizal fungi and other members of rhizosphere microbiota and discusses current developments and future trends concerning the following topics: (i) effect of soil microorganisms on mycorrhiza formation; (ii) mycorrhizosphere establishment; (iii) interactions involved in nutrient cycling and plant growth; (iv) interactions involved in the biological control of plant pathogens; and (v) interactions to improve soil quality . The main conclusion is that microbial interactions in the rhizosphere of mycorrhizal plants improve plant fitness and soil quality, critical issues for a sustainable agricultural development and ecosystem functioning. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 293 - 308 Significance of predation by protists in aquatic microbial food webs; Sherr EB et al.; Predation in aquatic microbial food webs is dominated by phagotrophic protists, yet these microorganisms are still understudied compared to bacteria and phytoplankton . In pelagic ecosystems, predaceous protists are ubiquitous, range in size from 2 gm flagellates to > 100 microm ciliates and dinoflagellates, and exhibit a wide array of feeding strategies . Their trophic states run the gamut from strictly phagotrophic, to mixotrophic: partly autotrophic and partly phagotrophic, to primarily autotrophic but capable of phagotrophy . Protists are a major source of mortality for both heterotrophic and autotrophic bacteria . They compete with herbivorous meso- and macro-zooplankton for all size classes of phytoplankton . Protist grazing may affect the rate of organic sinking flux from the euphotic zone . Protist excretions are an important source of remineralized nutrients, and of colloidal and dissolved trace metals such as iron, in aquatic systems . Work on predation by protists is being facilitated by methodological advances, e.g., molecular genetic analysis of protistan diversity and application of flow cytometry to study population growth and feeding rates . Examples of new research areas are studies of impact of protistan predation on the community structure of prey assemblages and of chemical communication between predator and prey in microbial food webs. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 189 - 95 Origins and diversification of sulfate-respiring microorganisms; Stahl DA et al.; If the diversification of microbial life can be depicted as a single tree, as inferred by comparative sequencing of ribosomal RNAs, this could provide a framework for defining the order of emergence of new metabolic pathways . However, recent recognition that lateral gene transfer has been a significant force in microbial evolution has created uncertainty about the interpretation of taxonomies based on gene sequences . In this context, the origins and evolution of sulfate respiration will be evaluated considering the evolutionary history of a central enzyme in this process, the dissimilatory sulfite reductase . These studies suggest at least two major lateral transfer events during the early diversification of sulfate respiring microorganisms . The high sequence conservation of this enzyme has also provided a mechanism to directly explore the natural diversity of sulfate-respiring organisms using molecular techniques, avoiding the bias of culture-based identification . These studies suggest that the habitat range and evolutionary diversity of this key functional group of organisms is greater than now appreciated. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 117 - 33 Principles of the light-limited chemostat: theory and ecological applications; Huisman J et al.; Light is the energy source that drives nearly all ecosystems on planet Earth . Yet, light limitation is still poorly understood . In this paper, we present an overview of the principles of the light-limited chemostat . The theory for light-limited chemostats differs considerably from the standard theory for substrate-limited chemostats . In particular, photons cannot be mixed by vigorous stirring, so that phototrophic organisms experience the light-limited chemostat as a heterogeneous environment . Similar to substrate-limited chemostats, however, light-limited chemostats do reach a steady state . This allows the study of phototrophic microorganisms under well-controlled light conditions, at a constant specific growth rate, for a prolonged time . The theory of the light-limited chemostat is illustrated with several examples from laboratory experiments, and a variety of ecological applications are discussed. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 107 - 15 Trade-offs and coexistence in microbial microcosms; Bohannan BJ et al.; Trade-offs among the abilities of organisms to respond to different environmental factors are often assumed to play a major role in the coexistence of species . There has been extensive theoretical study of the role of such trade-offs in ecological communities but it has proven difficult to study such trade-offs experimentally . Microorganisms are ideal model systems with which to experimentally study the causes and consequences of ecological trade-offs . In model communities of E . coli B and T-type bacteriophage, a trade-off in E . coli between resistance to bacteriophage and competitive ability is often observed . This trade-off can allow the coexistence of different ecological types of E . coli . The magnitude of this trade-off affects, in predictable ways, the structure, dynamics and response to environmental change of these communities . Genetic factors, environmental factors, and gene-by-environment interactions determine the magnitude of this trade-off . Environmental control of the magnitude of trade-offs represents one avenue by which environmental change can alter community properties such as invasability, stability and coexistence. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 85 - 97 Molecular biology of extremophiles: recent progress on the hyperthermophilic archaeon Sulfolobus; Ciaramella M et al.; Extremophiles are microorganisms that flourish in habitats of extreme temperature, pH, salinity, or pressure . All extreme environments are dominated by microorganisms belonging to Archaea, the third domain of life, evolutionary distinct from Bacteria and Eucarya . Over the past few years the biology of extremophilic Archaea has stimulated a lot of interest, aimed at understanding at molecular level the adaptation to their life conditions, as well as their evolutionary relationships to other organisms . Here, we review recent insights in the molecular biology of thermoacidophilic Archaea of the genus Sulfolobus, which has been used as a model system for biochemical, structural, and genetic studies in Archaea and extremophiles in general . With the recent completion of the genome sequence of Sulfolobus solfataricus it is expected that these organisms will contribute new discoveries in the near future. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 73 - 83 Novel thermoactive glucoamylases from the thermoacidophilic Archaea Thermoplasma acidophilum, Picrophilus torridus and Picrophilus oshimae; Serour E et al.; The thermoacidophilic Archaea Thermoplasma acidophilum (optimal growth at 60 degrees C and pH 1-2), Picrophilus torridus and Picrophilus oshimae (optimal growth at 60 degrees C and pH 0.7) were able to utilize starch as sole carbon source . During growth these microorganisms secreted heat and acid-stable glucoamylases into the culture fluid . Applying SDS gel electrophoresis activity bands were detected with appearent molecular mass (Mw) of 141.0, 95.0 kDa for T . acidophilum, 133.0, 90.0 kDa for P . torridus and 140.0, 85.0 kDa for P . oshimae . The purified enzymes were incubated with various polymeric substrates such as starch, pullulan, panose and isomaltose . The product pattern, analyzed by HPLC, showed that in all cases glucose was formed as the sole product of hydrolysis . The purified glucoamylases were optimally active at pH 2.0 and 90 degrees C and have an isoelectric points (pI) between 4.5 and 4.8 . Enzymatic activity was detected even at pH 1.0 and 100 degrees C . The glucoamylases were thermostable at elevated temperature with a half-life of 24 h at 90 degrees C for both P . torridus and T acidophilum, and 20 h at 90 degrees C for P oshimae . The enzyme system of T acidophilum has a lower Km value for soluble starch (1.06 mg/ml) than the enzymes from P . oshimae and P . torridus (4.35 mg/ml and 2.5 mg/ml), respectively . Enzyme activity was not affected by Na+, Mg++, Ca++, Ni++, Zn++, Fe++, EDTA and DTT. Mol Biol Evol, 2002 Dec, 19(12), 2265 - 76 Evolution of microbial genomes: sequence acquisition and loss; Berg OG et al.; We present models describing the acquisition and deletion of novel sequences in populations of microorganisms . We infer that most novel sequences are neutral . Thus, sequence duplications and gene transfer between organisms sharing the same environment are rarely expected to generate adaptive functions . Two classes of models are considered: (1) a homogeneous population with constant size, and (2) an island model in which the population is subdivided into patches that are in contact through slow migration . Distributions of gene frequencies are derived in a Moran model with overlapping generations . We find that novel, neutral or near-neutral coding sequences in microorganisms will not be fixed globally because they offer large target sizes for mutations and because the populations are so large . At most, such genes may have a transient presence in only a small fraction of the population . Consequently, a microbial population is expected to have a very large diversity of transient neutral gene content . Only sequences that are under strong selection, globally or in individual patches, can be expected to persist . We suggest that genome size is maintained in microorganisms by a quasi-steady state mechanism in which random fluctuations in the effective acquisition and deletion rates result in genome sizes that vary from patch to patch . We assign the genomic identity of a global population to those genes that are required for the participation of patches in the genetic sweeps that maintain the genomic coherence of the population . In contrast, we stress the influence of sequence loss on the isolation and the divergence (speciation) of novel patches from a global population. Med Hypotheses, 2002 Dec, 59(6), 626 - 9 The psychological puzzle presented by physiological acts; Seely S; Physiological acts present the puzzle that we are unable to explain how we performed them, how we solved the technical difficulties involved in their execution . For instance, when we sing, we produce various musical notes by changing the shape of the larynx . The normal person does not even know what the larynx looks like, let alone its configurations corresponding to various notes and the combination of the 19 muscles acting on the larynx that has to be activated to bring about the required configuration . Some psychologists offer the explanation that the technical details are worked out by a computer-like mechanism . But how could a computer start controlled cell proliferation in the repair of an injury? Another idea (originally put forward by Leibniz around 1700) is the complex character of the mind . The mind may incorporate a multitude of personalities . Subordinate personalities, some, perhaps, specialising on various tasks, may carry out the orders of the commanding personality . Objection: most physiological tasks, for instance digestion or the fight against invading microorganisms, are completely unconscious . The 'myself' personality could not even issue commands regarding the repair of injuries or the fight against microorganisms . PROPOSED SOLUTION: At an early stage of evolution the body of a higher animal divides itself into three compartments, each specialising on a different task of life . The three parts correspond to the entoderm, mesoderm and ectoderm appearing at an early part of ontogenetic development . The task of the entoderm is metabolism, that of the mesoderm mainly the mechanical aspects of life, that of the ectoderm gathering information from and reacting to the outside world . Each of these is probably of a composite character . The myself personality is the equivalent of the ectoderm, not of the whole body . The technical details of its commands are worked out by subordinate personalities . So are the commands of the mesodermal and entodermal divisions . Dreams could be attempts of communication between the germ layers . Clin Microbiol Infect, 2002 Nov, 8(11), 749 - 52 Detection of Brucella melitensis by the BacT/Alert automated system and Brucella broth culture; Ozkurt Z et al.; This study was conducted to evaluate the ability of the BacT/Alert automated blood culture system to detect Brucella spp . in comparison with traditional Brucella broth culture . Overall, 100 (50 bone marrow and 50 blood samples) paired cultures were obtained, and 59 were positive by at least one method . The Brucella broth culture method detected all 59 positive cultures (100%), and the BacT/Alert system detected 30 (50.8%) (P < 0.05) . The mean detection times for B . melitensis were 4.5 days in the BacT/Alert system and 5 days in Brucella broth culture (P > 0.05) . There is no significant difference between the two methods with respect to growth time of the microorganism, but Brucella broth culture is more sensitive than the BacT/Alert system. J Nat Prod, 2002 Nov, 65(11), 1683 - 4 A new carotenoid glycosyl ester isolated from a marine microorganism, Fusarium strain T-1; Sakaki H et al.; A new carotenoid glycosyl ester, neurosporaxanthin beta-D-glucopyranoside (2), together with neurosporaxanthin (1), beta-carotene, gamma-carotene, and torulene were isolated from cultured cells of a marine microorganism, strain T-1, which was identified as Fusarium sp . Their structures were determined by chemical and spectral data. Med Sci Monit, 2002 Nov, 8(11), BR454 - 9 Uric acid is a main electron donor to peroxidases in human blood plasma; Padiglia A et al.; BACKGROUND: Peroxidases are widely distributed and have been isolated from many higher-order plants, animal tissues, yeast and microorganisms . During measurements of peroxidase activities in samples of human plasma, we noticed the presence of a compound in the plasma which was interfering with the peroxidase assay . In this paper we describe the purification and characterization of this factor, which was identified as uric acid . MATERIAL/METHODS: The procedure used to purify uric acid from plasma involved ultra-filtration of the plasma, heat denaturation, DEAE-cellulose chromatography, and high performance liquid chromatography . The lyophilized powder was tested for homogeneity using an HPLC apparatus and capillary electrophoresis . Genuine uric acid samples were used for comparison . RESULTS: The compound obtained by the above-reported purification procedure was identified as uric acid by spectrophotometric analysis through comparison with genuine uric acid samples . Spectrophotometric measurements indicated that uric acid was degraded by HRP in the presence of H2O2 . CONCLUSIONS: The experimental procedures described above allowed us to isolate and identify uric acid as the component in human plasma that acts as a true substrate for peroxidases. Microbes Infect, 2002 Nov, 4(13), 1327 - 33 Human herpesvirus 6 and Chlamydia pneumoniae as etiologic agents in multiple sclerosis - a critical review; Swanborg RH et al.; Multiple sclerosis (MS) is thought by many investigators to have an infectious component, and several microorganisms have been associated with the disease during the last three decades . Recent studies have implicated both human herpesvirus 6 (HHV-6) and the obligate intracellular bacterium Chlamydia pneumoniae in the etiology of MS . As with earlier studies of other potential agents, however, evidence linking either of these organisms to the disease is equivocal . In this article, we review data for and against involvement of HHV-6 and C . pneumoniae in MS, as well as evidence concerning auxiliary factors, such as possession of the APOE epsilon4 allele, which may influence the role of these organisms in pathogenesis . Further, we suggest several lines of investigation that should clarify whether either or both pathogens are associated meaningfully with this disease. J Basic Microbiol, 2002, 42(6), 396 - 407 Diversity of microfungi in the phylloplane of plants growing in a Mediterranean ecosystem; Pereira PT et al.; Mediterranean ecosystems have not been investigated as natural habitats for microorganisms in general, and microfungi in particular . Plants harvested in "Serra da Arrabida" (38 degrees 27' N, 9 degrees 02' W), a Mediterranean ecosystem in Portugal, were analyzed for the filamentous microfungi inhabiting their surface . Two field locations with distinct climatic characteristics were studied: 'Fonte do Veado' (38 degrees 28'50" N, 9 degrees 0'17" W; 300 m elevation) located on the northern slope, and 'Mata do Solitario' (38 degrees 27'55" N, 8 degrees 59'35" W; 50 m elevation), on the southern slope . From Veado zone, leaf samples yielded a total of 3,049 isolates, ranging from 317 to 1,328/sample (mean = 762) . The number of species/sample ranged from 12 to 24 . From Solitario zone, leaf samples yielded a total of 1,337 isolates, ranging from 189 to 528/sample (mean = 334) . The number of species/sample, in this case, ranged from 10 to 17 . Veado zone showed a wider range of species . The fungal species more frequently isolated from both zones (Aureobasidium pullulans (De Bary) Arnaud, Cladosporium cladosporioides (Fresen.) De Vries, C . sphaerospermum Penzig and Alternaria alternata (Fr.) Keissler) were found in all plant samples and represents 80% (Veado) and 85% (Solitario) of the total isolates. Curr Opin Ophthalmol, 2002 Dec, 13(6), 393 - 6 Sarcoidosis; Jones NP; Sarcoidosis commonly involves the eye, causing uveitis, the lacrimal gland, and the cranial nerves, including the optic nerve itself . Several microorganism types have recently been located in sarcoid granulomas, suggesting an infective cause in predisposed individuals . The approach to diagnosis has been refined, including the use of high-resolution computed tomography of the chest . New ophthalmic manifestations have been described, suggesting a wider role for sarcoidosis in intraocular inflammation, and indocyanine green angiography has clarified choroidal involvement . Immunosuppressive and anticytokine treatments can be effective in severe systemic sarcoidosis and should be considered in sight-threatening disease. Protein Sci, 2002 Dec, 11(12), 2958 - 68 Cloning, overexpression, and characterization of a bacterial Ca2+-dependent phospholipase D; Yang H et al.; Phospholipase D (PLD), an important enzyme involved in signal transduction in mammals, is also secreted by many microorganisms . A highly conserved HKD motif has been identified in most PLD homologs in the PLD superfamily . However, the Ca(2+)-dependent PLD from Streptomyces chromofuscus exhibits little homology to other PLDs . We have cloned (using DNA isolated from the ATCC type strain), overexpressed in Escherichia coli (two expression systems, pET-23a(+) and pTYB11), and purified the S . chromofuscus PLD . Based on attempts at sequence alignment with other known Ca(2+)-independent PLD enzymes from Streptomyces species, we mutated five histidine residues (His72, His171, His187, His200, His226) that could be part of variants of an HKD motif . Only H187A and H200A showed dramatically reduced activity . However, mutation of these histidine residues to alanine also significantly altered the secondary structure of PLD . Asparagine replacements at these positions yielded enzymes with structure and activity similar to the recombinant wild-type PLD . The extent of phosphatidic acid (PA) activation of PC hydrolysis by the recombinant PLD enzymes differed in magnitude from PLD purified from S . chromofuscus culture medium (a 2-fold activation rather than 4-5-fold) . One of the His mutants, H226A, showed a 12-fold enhancement by PA, suggesting this residue is involved in the kinetic activation . Another notable difference of this bacterial PLD from others is that it has a single cysteine (Cys123); other Streptomyces Ca(2+)-independent PLDs have eight Cys involved in intramolecular disulfide bonds . Both C123A and C123S, with secondary structure and stability similar to recombinant wild-type PLD, exhibited specific activity reduced by 10(-5) and 10(-4) . The Cys mutants still bound Ca(2+), so that it is likely that this residue is part of the active site of the Ca(2+)-dependent PLD . This would suggest that S . chromofuscus PLD is a member of a new class of PLD enzymes. Biochem Soc Trans, 2002 Nov, 30(Pt 6), 1006 - 10 Structure and function of complement C5 convertase enzymes; Pangburn MK et al.; The multisubunit enzymes of the complement system that cleave C5 have many unusual properties, the most striking of which is that they acquire their specificity for C5 following cleavage of another substrate C3 . C5 convertases are assemblies of two proteins C4b and C2a (classical or lectin pathways) or C3b and Bb (alternative pathway) and additional C3b molecules . The catalytic complexes (C4b, C2a or C3b, Bb) are intrinsically unstable ( t (1/2)=1-3 min) and the enzymes are controlled by numerous regulatory proteins that accelerate this natural decay rate . Immediately after assembly, the bi-molecular enzymes preferentially cleave the protein C3 and exhibit poor activity toward C5 (a K (m) of approx . 25 microM and a C5 cleavage rate of 0.3-1 C5/min at V (max)) . Efficient C3 activation results in the covalent attachment of C3b to the cell surface and to the enzyme itself, resulting in formation of C3b-C3b and C4b-C3b complexes . Our studies have shown that deposition of C3b alters the specificity of the enzymes of both pathways by changing the K (m) for C5 more than 1000-fold from far above the physiological C5 concentration to far below it . Thus, after processing sufficient C3 at the surface of a microorganism, the enzymes switch to processing C5, which initiates the formation of the cytolytic membrane attack complex of complement. Cell Mol Life Sci, 2002 Sep, 59(9), 1428 - 59 The superoxide-generating NADPH oxidase: structural aspects and activation mechanism; Vignais PV; Flavocytochrome b558 is the catalytic core of the respiratory-burst oxidase, an enzyme complex that catalyzes the NADPH-dependent reduction of O2 into the superoxide anion O2 in phagocytic cells . Flavocytochrome b558 is anchored in the plasma membrane . It is a heterodimer that consists of a large glycoprotein gp91phox (phox forphagocyte oxidase) (beta subunit) and a small protein p22phox (alpha subunit) . The other components of the respiratory-burst oxidase are water-soluble proteins of cytosolic origin, namely p67phox, p47phox, p40phox and Rac . Upon cell stimulation, they assemble with the membrane-bound flavocytochrome b558 which becomes activated and generates O2- . A defect in any of the genes encoding gp91phox, p22phox, p67phox or p47phox results in chronic granulomatous disease, a genetic disorder characterized by severe and recurrent infections, illustrating the role of O2- and the derived metabolites H2O2 and HOCl in host defense against invading microorganisms . The electron carriers, FAD and hemes b, and the binding site for NADPH are confined to the gp91phox subunit of flavocytochrome b558 . The p22phox subunit serves as a docking site for the cytoso lic phox proteins . This review provides an overview of current knowledge on the structural organization of the O2(-)-generating flavocytochrome b558, its kinetics, its mechanism of activation and the regulation of its biosynthesis . Homologues of gp91phox, called Nox and Duox, are present in a large variety of non-phagocytic cells . They exhibit modest O2(-)-generating oxidase activity, and some act as proton channels . Their role in various aspects of signal transduction is currently under investigation and is briefly discussed. Proc Natl Acad Sci U S A, 2002 Nov 26, 99(24), 15681 - 6 Epub 2002 Nov 18. Cultivating the uncultured; Zengler K et al.; The recent application of molecular phylogeny to environmental samples has resulted in the discovery of an abundance of unique and previously unrecognized microorganisms . The vast majority of this microbial diversity has proved refractory to cultivation . Here, we describe a universal method that provides access to this immense reservoir of untapped microbial diversity . This technique combines encapsulation of cells in gel microdroplets for massively parallel microbial cultivation under low nutrient flux conditions, followed by flow cytometry to detect microdroplets containing microcolonies . The ability to grow and study previously uncultured organisms in pure culture will enhance our understanding of microbial physiology and metabolic adaptation and will provide new sources of microbial metabolites . We show that this technology can be applied to samples from several different environments, including seawater and soil. Ann N Y Acad Sci, 2002 Oct, 971, 359 - 61 Structural and biological characterization of chromofungin, the antifungal chromogranin A (47-66)-derived peptide; Lugardon K et al.; The antifungal peptide named chromofungin is the most active vasostatin-I-derived peptide, corresponding to the sequence 47-66 of chromogranin A . (1)H-NMR analysis revealed that it adopts a helical structure . The mechanism implicated in the interaction of chromofungin with fungi and yeast cells was studied by penetration of monolayers and confocal laser microscopy . Chromofungin is able to interact with the cell wall, to cross the plasma membrane, to accumulate in the microorganism, and to inhibit calcineurin activity. Biomarkers, 2002 Sep-Oct, 7(5), 347 - 421 Biomonitoring of arylamines and nitroarenes; Sabbioni G et al.; Arylamines and nitroarenes are very important intermediates in the industrial manufacture of dyes, pesticides and plastics, and are significant environmental pollutants . The metabolic steps of N-oxidation and nitroreduction to yield N-hydroxyarylamines are crucial for the toxic properties of arylamines and nitroarenes . Nitroarenes are reduced by microorganisms in the gut or by nitroreductases and aldehyde dehydrogenase in hepatocytes to nitrosoarenes and N-hydroxyarylamines . N-Hydroxyarylamines can be further metabolized to N-sulphonyloxyarylamines, N-acetoxyarylamines or N-hydroxyarylamine N-glucuronide . These highly reactive intermediates are responsible for the genotoxic and cytotoxic effects of this class of compounds . N-Hydroxyarylamines can form adducts with DNA, tissue proteins, and the blood proteins albumin and haemoglobin in a dose-dependent manner . DNA and protein adducts have been used to biomonitor humans exposed to such compounds . All these steps are dependent on enzymes, which are present in polymorphic forms . This article reviews the metabolism of arylamines and nitroarenes and the biomonitoring studies performed in animals and humans exposed to these substances. Biotechnol Annu Rev, 2002, 8, 227 - 65 Antibiotics; Behal V; The chapter informs about different types of antibiotics, their structure, biosynthesis and their regulation . Industrial cultivation and isolation of antibiotics is described in the chapter . Search for microorganisms producing antibiotics and preparation of high-producing strains is described . Resistance against antibiotics in producing microorganisms and pathogens is discussed. Mikrobiol Z, 2002 Jul-Aug, 64(4), 50 - 61 {Infection of cell cultures by mollicutes: disease symptoms, means and methods of its detection and treatment}; Skrypal' IH; Up-to-date data concerning the cell cultures infection by mollicutes have been generalized in the paper in a brief but maximum complete form . Different symptoms of such infection as well as the changes taking place in the cell cultures under the effect of various types of lesion--from latent to acute have been described . The methods of mollicutes detection in cell cultures, methods of their release of these microorganisms have been stated in detail . The work is called to help specialists who work with cell cultures, use them in biotechnological production or keep their collections. Pathologe, 2002 Nov, 23(6), 465 - 71 Epub 2002 Oct 08. {Differentiation of granulomatous lesions in the bone marrow}; Kvasnicka HM et al.; Granulomas are an infrequent finding in bone marrow biopsies and may be associated with a broad spectrum of infectious and non-infectious disorders . In this context sarcoidosis generally reveals the highest incidence of such bone marrow lesions . Other granulomas may be encountered in drug-induced secondary changes (toxic myelopathy) and in particular also in HIV myelopathy . In these cases the granulomas are small, ill-defined and difficult to recognize . Special staining methods are required to detect microorganisms . Fibrin ring granulomas (doughnut granulomas) are typical for Q-fever, but may also be seen in reactive conditions, after drug therapy and in the course of other infectious disorders, such as Lyme disease . Therefore a broad serological testing should be performed . In conclusion, in most cases the morphological findings in the bone marrow are not specific so that a synoptical approach regarding histological, clinical and serological data is warranted to reach the correct diagnosis. FEMS Microbiol Lett, 2002 Nov 5, 216(2), 179 - 83 Overexpression in Escherichia coli of the AT-rich trpA and trpB genes from the hyperthermophilic archaeon Pyrococcus furiosus; Ishida M et al.; Expression of AT-rich genes from microorganisms such as archaea is often inefficient in Escherichia coli . The trpA and trpB genes encoding the tryptophan synthase subunits were cloned from the hyperthermophilic archaeon Pyrococcus furiosus . No apparent difference in codon bias was found between the genes . However, using a conventional cloning vector having the lac promoter, the trpB gene was expressed poorly in E . coli, whereas the trpA gene was overexpressed . The expression of the trpB gene was remarkably enhanced (>12-fold) by the introduction of an overlapping leader open reading frame . The expression of the trpA gene was also improved ( approximately 1.5-fold) . This approach may be useful for overexpressing various kinds of AT-rich genes. FEMS Microbiol Lett, 2002 Nov 5, 216(2), 123 - 31 Microbial aldo-keto reductases; Ellis EM; The aldo-keto reductases (AKR) are a superfamily of enzymes with diverse functions in the reduction of aldehydes and ketones . AKR enzymes are found in a wide range of microorganisms, and many open reading frames encoding related putative enzymes have been identified through genome sequencing projects . Established microbial members of the superfamily include the xylose reductases, 2,5-diketo-D-gluconic acid reductases and beta-keto ester reductases . The AKR enzymes share a common (alpha/beta)(8) structure, and conserved catalytic mechanism, although there is considerable variation in the substrate-binding pocket . The physiological function of many of these enzymes is unknown, but a variety of methods including gene disruptions, heterologous expression systems and expression profiling are being employed to deduce the roles of these enzymes in cell metabolism . Several microbial AKR are already being exploited in biotransformation reactions and there is potential for other novel members of this important superfamily to be identified, studied and utilized in this way. Carbohydr Res, 2002 Nov 19, 337(21-23), 2017 - 22 A convenient synthesis of C-galactofuranosylic compounds (C-galactofuranosides); Owen DJ et al.; Galactofuranose sugar units are essential for the production of the cell coat of many pathogenic microorganisms . This sugar is not found in mammals, and so compounds that may interfere with the biosynthetic processing of this sugar unit provide interesting targets for drug design . This paper describes the use of a cyanation reaction for the production of a one-carbon extension of a galactofuranosylic unit at C-1, giving 2,5-anhydro-3,4,6,7-tetra-O-benzoyl-D-glycero-L-manno-heptononitrile . A procedure for the efficient hydrolysis of the introduced nitrile group to produce the methyl ester is reported, along with procedures for the synthesis of both the corresponding alpha,beta-unsaturated, and 3-deoxy ester derivatives . Anal Chem, 2002 Nov 1, 74(21), 5523 - 30 Mechanistic aspects in the generation of apparent ultrahigh efficiencies for colloidal (microbial) electrokinetic separations; Armstrong DW et al.; Under specific experimental conditions, the electrokinetic separation of certain microorganisms can produce peaks of very high apparent efficiencies (approximately 10(6)-10(10) theoretical plates/m) . This is unusual in that no deliberate focusing mechanism was employed . To investigate this process further, the separation was monitored in real time using a charge-coupled device (CCD) imaging system . At least two different processes seem to be operative when these narrow peaks are observed . The initial field-induced association of cells appears to require a dilute polymer solution, electroosmotic flow (preferably countercurrent to the direction of cell electrophoresis), and a direct current electric field . Three possible models are presented that may explain aspects of the observed behavior . The balance between dispersive forces and intercellular adhesive forces also affects the observed bandwidths . Understanding and controlling the dynamic and aggregation of cells in microfluidic processes is essential, since it can be beneficial for some experiments and detrimental to others. Radiographics, 2002 Nov-Dec, 22(6), 1411 - 28 Neuroimaging in pediatric leukemia and lymphoma: differential diagnosis; Vazquez E et al.; Recent advances in therapy for pediatric hematologic neoplasms have greatly improved the prognosis but have resulted in an increased incidence of associated complications and toxic effects . The main neuroimaging features in pediatric patients with leukemia or lymphoma treated with chemotherapy or radiation therapy were retrospectively reviewed . To simplify the approach and facilitate differential diagnosis, the neuroimaging features have been classified into three main categories: central nervous system manifestations of primary disease, side effects of therapeutic procedures (radiation therapy, chemotherapy, bone marrow transplantation), and complications due to immunosuppression, particularly infections . Manifestations of primary disease include cerebrovascular complications (hemorrhage, cerebral infarction) and central nervous system involvement (infiltration of the meninges, parenchyma, bone marrow, orbit, and spine) . Effects of radiation therapy include white matter disease, mineralizing microangiopathy, parenchymal brain volume loss, radiation-induced cryptic vascular malformations, and second neoplasms . Effects of chemotherapy and bone marrow transplantation include hemorrhage, dural venous thrombosis, white matter disease, reversible posterior leukoencephalopathy syndrome, and anterior lumbosacral radiculopathy . Both the underlying malignancy and antineoplastic therapy can cause immunosuppression . Fungi are the most frequent causal microorganisms in immunosuppressed patients with infection . Familiarity with the imaging findings is essential for proper diagnosis of neurologic symptoms in pediatric patients with oncohematologic disease . Copyright RSNA, 2002 J Eur Acad Dermatol Venereol, 2002 Sep, 16(5), 438 - 40 Manifestation of cellulitis after saphenous venectomy for coronary bypass surgery; Karakas M et al.; A few cases of onset of cellulitis after saphenous venectomy for coronary by-pass surgery were first reported by Baddour and Bisno in 1982 . We reviewed the dinical characteristics of 31 subjects followed up in our department following onset of manifestations of cellulitis after saphenous venectomy for coronary by-pass surgery . In all the subjects the cellulitis originated at the scar of the saphenous venectomy, and most presented ill-defined, mildly erythematous, slightly oedematous lesions . Mycologically confirmed tinea pedis was found in 25 subjects . All the patients responded well to penicillins or cephalosporins . In this report we comment on the pathogenesis of this complication of saphenous venectomy based on the clinical features of the cases reviewed . We suggest that saphenous venectomy may destroy the lymphatics in the lower leg, that are located adjacent to the great saphenous vein, and thus microorganisms penetrating into the skin may easily cause cellulitis in areas with impaired lymphatic drainage. Cas Lek Cesk, 2002 Sep 22, 141 Suppl, 23 - 5 {Vaccinia virus as a vector for transduction of dendritic cells}; Nemeckova S et al.; Dendritic cells (DC) are very heterogenous population of professional antigen-presenting cells . Precursor cells migrate from bone marrow to peripheral tissues, where immature DC ingest pathogenic microorganisms and then migrate to secondary lymphoid organs . DC differentiate into mature cells that are capable to prime naive T lymphocytes . DC can be used for immunotherapy of cancer and infectious diseases . Transduction of DC by recombinant viral vectors expressing tumor associated antigens (TAA) can result in efficient antigen presentation to T lymphocytes . DC transduced with recombinant vaccinia virus expressing E7 oncoprotein of human papilloma virus 16 are able to protect mice against the growth of syngeneic papillomavirus transformed tumor cells TC1 . Antitumor effect was observed also with nonreplicating viruses. Nihon Kokyuki Gakkai Zasshi, 2002 Aug, 40(8), 703 - 7 {A case of mixed infection by Nocardia asteroides and Mycobacterium tuberculosis}; Shimokubo T et al.; A 55-year-old man with diabetes mellitus was admitted to our hospital because of abnormal shadows in his chest radiographs . Both chest radiography and CT revealed infiltrative shadows in the right upper lung field . Repeated sputum smears showed no mycobacterium, so bronchoalveolar lavage (BAL) was performed bronchoscopically at the right B3b . The BAL fluid and the sputum obtained on the day after BAL contained acid-fast, branching filamentous structures . The microorganism was identified as Nocardia asteroides . Trimethoprim-sulfamethoxazole (ST) and SPFX were therefore administered . Later, Mycobacterium tuberculosis was detected in a 6-week culture of the sputum and BAL fluid . This case was diagnosed as a mixed infection by Nocardia asteroides and Mycobacterium tuberculosis, so the three anti-tuberculosis agents INH, RFP and EB were added . After 6 months of the combined therapy, neither microorganism could be detected in the sputum, and the lesion in the CT scan had decreased markedly in size . Since such a mixed infection is very rare, no treatment strategy has yet been established . The combined therapy was judged to have been effective in this case. Pharmacoepidemiol Drug Saf, 2002 Sep, 11(6), 447 - 53 Changes in prescribed drug doses after market introduction; Heerdink ER et al.; PURPOSE: The establishment of recommended dosing regimens has always been a difficult aspect of drug development . This paper examines the extent to which postmarketing prescribing deviates from initially recommended dosing regimens . We used the World Health Organization's (WHO) periodically updated compilation of the 'Defined Daily Dose' (DDD) to reflect prevailing patterns of prescribing in national markets . The aim of this study was to evaluate DDD changes over time (1982-2000) and to identify possible determinants of these changes . METHODS: Data on DDD changes were obtained from the WHO's Oslo Collaborating Centre . We performed a case-control analysis in which we compared drugs with (cases) and without (controls) postmarketing changes in DDD on possible determinants associated with DDD change . RESULTS: We found 115 instances of a change of DDD in the period 1982-2000 (45 (39.1%) increases and 70 (60.9%) decreases) . Antibiotics showed the greatest number of changes in DDD: predominantly increases in the 1980s, while the 1990s were dominated by decreases in DDD of mostly cardiovascular drugs . CONCLUSION: Changes in DDD reflect the outcome of a melange of forces, including misconceptions of dose requirements during pre-market development of drug and postmarketing changes in pharmacotherapeutic knowledge, clinical concepts, economic forces, and, in the case of anti-infective agents, changing patterns of resistance/sensitivity of target microorganisms to the anti-infective agent(s) in question. Environ Health Perspect, 2002 Oct, 110 Suppl 5, 807 - 11 Signaling from toxic metals to NF-kappaB and beyond: not just a matter of reactive oxygen species; Chen F et al.; The nuclear factor kappa B (NF-kappaB) family of transcription factors controls expression of a number of early response genes associated with inflammatory responses, cell growth, cell cycle progression, and neoplastic transformation . These genes include a multitude of cytokines, chemokines, adhesion molecules, immune receptors, stress proteins, apoptotic or anti-apoptotic regulators, and several oncogenes . Accumulating evidence indicates that a variety of toxic metals are able to affect the activation or activity of NF-kappaB, but the molecular mechanisms involved in this process remain largely unknown . The signaling pathways mediating cytokine- or microorganism-induced NF-kappaB activation have been well established recently . Whether the same signaling systems are involved in metal-induced NF-kappaB activation, however, is unclear . In the present review, we have attempted to evaluate and update the possible mechanisms of metal signals on the activation and function of NF-kappaB. Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet, 2001, 66(2a), 249 - 56 The physiological development of the chlorotic lesion induced by coronatine; Harzallah D et al.; Toxins are secondary metabolites produced by several plant pathogenic microorganisms . These toxins play a major role in the development of disease symptoms . Coronatine, the non specific toxin, was extracted and purified from the culture medium of Pseudomonas syringae pv . glycinea . In this study, the effect of coronatine on the development of the chlorotic lesion on the Phaseolus vulgaris L . leaves, indicates that coronatine induced chlorosis on treated leaves as well as or untreated leaves on the same plant . An other effect of this toxin is the reduction of amount of both chlorophyll a and b . These results, provide that the development of chlorotic lesion is a primarily mode of action of coronatine. J Clin Lab Anal, 2002, 16(6), 304 - 7 Changes in the mannan binding lectin (MBL) concentration in human milk during lactation; Tregoat V et al.; The mannan binding lectin (MBL) activates the complement system by the lectin pathway after the recognition of some structural motifs (saccharides) present on the surface of microorganisms . MBL has been mostly identified and quantified in human serum by ELISA or microparticle immunonephelometry assays . This article reports the MBL levels as assessed by a microparticle immunonephelometric assay in 76 human milk samples . Immunonephelometry was performed using skim-milk samples diluted 20 times over a calibration range of 0.07-4.82 mg/L . MBL is indeed present in human milk and its concentration decreases significantly during development from colostrum (0.55+/-0.09 mg/L) to transitional (0.18+/-0.02 mg/L) and mature milk (0.17+/-0.02 mg/L) . This innate molecule may be involved in the primary defenses of the mammary gland and the neonate, whose immune system is immature . The high levels observed during the first days of lactation support the hypothesis that this molecule plays a key role in limiting the colonization of the newborn gut by pathogens . J Biol Chem, 2003 Jan 17, 278(3), 1561 - 8 Epub 2002 Nov 06. Minimally modified LDL binds to CD14, induces macrophage spreading via TLR4/MD-2, and inhibits phagocytosis of apoptotic cells; Miller YI et al.; Minimally modified low density lipoprotein (mmLDL) is a pro-inflammatory and pro-atherogenic lipoprotein that, unlike profoundly oxidized LDL (OxLDL), is not recognized by scavenger receptors and thus does not have enhanced uptake by macrophages . However, here we demonstrate that mmLDL (as well as OxLDL) induces actin polymerization and spreading of macrophages, which results in such pro-atherogenic consequences as inhibition of phagocytosis of apoptotic cells but enhancement of OxLDL uptake . We also demonstrate for the first time that the lipopolysaccharide receptor, CD14, and toll-like receptor-4/MD-2 are involved in these mmLDL effects . Macrophages of the J774 cell line exhibited higher mmLDL binding and F-actin response than its CD14-deficient mutant, LR-9 cells . Similarly, Chinese hamster ovary cells transfected with human CD14 specifically bound mmLDL and responded with higher F-actin compared with control cells . Macrophages from C3H/HeJ mice, which have a point mutation in the Tlr4 gene, responded with lower F-actin to mmLDL and did not spread as well as macrophages from control animals . A significantly higher F-actin response was also observed in Chinese hamster ovary cells transfected with human toll-like receptor-4/MD-2 but not with TLR4 alone or TLR2 . Thus, in addition to inhibition of phagocytosis, the recognition of mmLDL by macrophage lipopolysaccharide receptors results in convergence of cellular immune responses to products of microorganisms and to oxidation-specific self-antigens, which could both influence macrophage function and atherogenesis. FEMS Microbiol Lett, 2002 Oct 29, 216(1), 15 - 21 Competitive hybridization on spotted microarrays as a tool to conduct comparative genomic analyses of Xylella fastidiosa strains; Costa de Oliveira R et al.; Xylella fastidiosa strains are responsible for several plant diseases and since such isolates display a broad host range and complex biological behavior, genomic comparisons employing microarray hybridizations may provide an effective method to compare them . Thus, we performed a thorough validation of this type of approach using two recently sequenced strains of this phytopathogen . By matching microarray hybridization results to direct sequence comparisons, we were able to establish precise cutoff ratios for common and exclusive sequences, allowing the identification of exclusive genes involved in important biological traits . This validation will enable the use of microarray-based comparisons across a wide variety of microorganisms Mol Plant Microbe Interact, 2002 Nov, 15(11), 1165 - 72 Fungal ABC transporters and microbial interactions in natural environments; Schoonbeek HJ et al.; In natural environments, microorganisms are exposed to a wide variety of antibiotic compounds produced by competing organisms . Target organisms have evolved various mechanisms of natural resistance to these metabolites . In this study, the role of ATP-binding cassette (ABC) transporters in interactions between the plant-pathogenic fungus Botrytis cinerea and antibiotic-producing Pseudomonas bacteria was investigated in detail . We discovered that 2,4-diacetylphloroglucinol, phenazine-1-carboxylic acid and phenazine-1-carboxamide (PCN), broad-spectrum antibiotics produced by Pseudomonas spp., induced expression of several ABC transporter genes in B . cinerea . Phenazines strongly induced expression of BcatrB, and deltaBcatrB mutants were significantly more sensitive to these antibiotics than their parental strain . Treatment of B . cinerea germlings with PCN strongly affected the accumulation of {14C}fludioxonil, a phenylpyrrole fungicide known to be transported by BcatrB, indicating that phenazines also are transported by BcatrB . Pseudomonas strains producing phenazines displayed a stronger antagonistic activity in vitro toward ABcatrB mutants than to the parental B . cinerea strain . On tomato leaves, phenazine-producing Pseudomonas strains were significantly more effective in reducing gray mold symptoms incited by a ABcatrB mutant than by the parental strain . We conclude that the ABC transporter BcatrB provides protection to B . cinerea in phenazine-mediated interactions with Pseudomonas spp . Collectively, these results indicate that fungal ABC transporters can play an important role in antibiotic-mediated interactions between bacteria and fungi in plant-associated environments . The implications of these findings for the implementation and sustainability of crop protection by antagonistic microorganisms are discussed. J Immunol, 2002 Nov 15, 169(10), 5726 - 34 CL-46, a novel collectin highly expressed in bovine thymus and liver; Hansen S et al.; Collectins are oligomeric molecules with C-type lectin domains attached to collagen-like regions via alpha-helical neck regions . They bind nonself glycoconjugates on the surface of microorganisms and inhibit infection by direct neutralization, agglutination, or opsonization . During the characterization of the gene encoding bovine CL-43 (43-kDa collectin), we identified a novel collectin-gene . We report the cloning and partial characterization of the novel collectin CL-46 . The mRNA comprises 1188 nucleotides encoding a protein of 371 aa with an included leader peptide of 20 residues . CL-46 has two cysteine residues in the N-terminal segment, a potential N-glycosylation site in the collagen region, and an extended hydrophilic loop close to the binding site of the carbohydrate recognition domain . It is expressed in the thymus, liver, mammary gland, and tissues of the digestive system . Recombinant CL-46 corresponding to the alpha-helical neck region and the C-type lectin domain binds preferential N-acetyl-D-glucoseamine and N-acetyl-D-mannoseamine . The gene encoding CL-46 spans approximately 10 kb and consists of eight exons, with high structural resemblance to the gene encoding human surfactant protein D . It is located on the bovine chromosome 28 at position q1.8 together with the gene encoding conglutinin and CL-43 . Several potential thymus-related cis-regulatory elements were identified in the 5'-upstream sequence, indicating that the expression in thymus may be modulated by signals involved in T cell development. J Biomed Opt, 2002 Oct, 7(4), 546 - 54 Biocompatible semiconductor optoelectronics; Gourley PL et al.; We investigate optoelectronic properties of integrated structures comprising semiconductor light-emitting materials for optical probes of microscopic biological systems . Compound semiconductors are nearly ideal light emitters for probing cells and other microorganisms because of their spectral match to the transparency wavelengths of biomolecules . Unfortunately, the chemical composition of these materials is incompatible with the biochemistry of cells and related biofluids . To overcome these limitations, we investigate functionalized semiconductor surfaces and structures to simultaneously enhance light emission and the flow of biological fluids in semiconductor microcavities . We have identified several important materials problems associated with the semiconductor/biosystem interface . One is the biofluid degradation of electroluminescence by ionic diffusion into compound semiconductors . Ions that diffuse into the active region of a semiconductor light emitter can create point defects that degrade the quantum efficiency of the radiative recombination process . In this paper we discuss ways of mitigating these problems using materials design and surface chemistry, and suggest future applications for these materials. Water Res, 2002 Oct, 36(17), 4227 - 34 Fluorescent dye labeled bacteriophages--a new tracer for the investigation of viral transport in porous media: 1 . Introduction and characterization; Gitis V et al.; A new method for the study of pathogen transport in porous media is presented . The method is based on conjugation of fluorescent dyes to target bacteriophages and application of the modified bacteriophages for tracer studies . We demonstrate that the relevant transport determining properties of Rhodamine and several fluorescein-labeled phages are practically identical to those of the native bacteriophages . The advantages of the proposed method relative to direct enumeration of bacteriophages by plaque forming unit method, turbidity, fluorescent microspheres, and other alternative tracers are discussed . Notable advantages include simple quantitation by optical methods, unbiased signals even when virus aggregates are formed, and the ability to decouple inactivation kinetics from transport phenomena . Additionally, the signal reflects the removal and transport of the studied microorganism and not a surrogate. J Oral Maxillofac Surg, 2002 Nov, 60(11), 1234 - 40 Microbial complexes detected in the second/third molar region in patients with asymptomatic third molars; White RP Jr et al.; PURPOSE: Our goal was to report the detection and levels of pathogenic bacteria in subgingival plaque samples taken from the distal of all second molars in 295 patients with asymptomatic third molars . PATIENTS AND METHODS: Data assessing oral health were collected from each of these healthy patients (ASA Classes I and II) . Probing depth (PD), at 6 sites per tooth, including third molars, was obtained to determine periodontal status . Subgingival plaque samples were taken from the distal of all second molars before periodontal probing . The presence and levels of 11 bacterial species were determined using whole chromosomal DNA probes and checkerboard DNA-DNA hybridization . Detected bacterial species were grouped into clusters of periodontal pathogens designated as "red" or "orange" complex microorganisms as described by Socransky et al (J Clin Periodontal 25:134, 1998) who found an association of these specific microorganisms with periodontitis . RESULTS: As a group these relatively young patients were periodontally healthy . "Orange and red" complex microorganisms were detected at levels equal to or greater than 10(5) more often if patients had a PD equal to or greater than 5 mm with periodontal attachment loss at the distal of second molars or around third molars at their entry examination . In patients with no PD equal to or greater than 5 mm in the third molar region, "orange and red" complex microorganisms were detected at levels equal to or greater than 10(5) more frequently than would be anticipated in patients with little clinical evidence of periodontal disease . CONCLUSIONS: The clinical findings of increased periodontal PDs and periodontal attachment loss coupled with colonization of periodontal pathogens support the concept that clinical and microbial changes associated with the initiation of periodontitis may present first in the third molar region in young adults . Trends Microbiol, 2002 Nov, 10(11), 483 - 4; discussion 484-5 The use of animal infection models to study the pathogenesis of melioidosis and glanders; Woods DE; The use of animal infection models is central to the study of microbial pathogenesis . In combination with genetic, immunological and antigen purification techniques, much can be learned regarding the pathogenesis of diseases caused by microorganisms . This update focuses on the recent use of animal infection models to study the pathogenesis of melioidosis and glanders. Curr Biol, 2002 Oct 29, 12(21), R731 - 2 Innate immunity: the worm fights back; Nicholas HR et al.; Innate immunity is an evolutionarily ancient defense system that enables animals and plants to resist invading microorganisms . Recent studies have demonstrated the existence of innate immune responses in Caenorhabditis elegans. Acta Odontol Scand, 2002 Oct, 60(5), 257 - 64 Effect of pregnancy on periodontal and dental health; Laine MA; Clinical studies have shown that oral tissues can be affected by pregnancy . Pregnancy-related changes are most frequent and most marked in gingival tissue . Pregnancy does not cause gingivitis, but may aggravate pre-existing disease . The most marked changes are seen in gingival vasculature . Characteristic of pregnancy gingivitis is that the gingiva is dark red, swollen, smooth and bleeds easily . Women with pregnancy gingivitis may sometimes develop localized gingival enlargements . The gingival changes usually resolve within a few months of delivery if local irritants are eliminated . The inflammatory changes are usually restricted to the gingiva and probably do not cause permanent changes in periodontal tissues more often than those in the non-pregnant state . Although it is widely believed that pregnancy is harmful to the teeth, the effect of pregnancy on the initiation or progression of caries is not clear . Previous studies, however, indicate that the teeth do not soften, i.e . no significant withdrawal of calcium or other minerals occurs in the teeth . It is mainly the environment of the tooth that is affected . The number of certain salivary cariogenic microorganisms may increase in pregnancy, concurrently with a decrease in salivary pH and buffer effect . Changes in salivary composition in late pregnancy and during lactation may temporarily predispose to dental caries and erosion . Although their underlying mechanisms of action are not fully understood, pregnancy-related changes in the oral environment may have some untoward temporary or permanent effects on oral health . Most of these effects could be avoided by practising good oral hygiene. MMWR Recomm Rep, 2002 Oct 25, 51(RR-16), 1 - 45, quiz CE1-4 Guideline for Hand Hygiene in Health-Care Settings . Recommendations of the Healthcare Infection Control Practices Advisory Committee and the HICPAC/SHEA/APIC/IDSA Hand Hygiene Task Force . Society for Healthcare Epidemiology of America/Association for Professionals in Infection Control/Infectious Diseases Society of America; Boyce JM et al.; The Guideline for Hand Hygiene in Health-Care Settings provides health-care workers (HCWs) with a review of data regarding handwashing and hand antisepsis in health-care settings . In addition, it provides specific recommendations to promote improved hand-hygiene practices and reduce transmission ofpathogenic microorganisms to patients and personnel in health-care settings . This report reviews studies published since the 1985 CDC guideline (Garner JS, Favero MS . CDC guideline for handwashing and hospital environmental control, 1985 . Infect Control 1986;7:231-43) and the 1995 APIC guideline (Larson EL, APIC Guidelines Committee . APIC guideline for handwashing and hand antisepsis in health care settings . Am J Infect Control 1995;23:251-69) were issued and provides an in-depth review of hand-hygiene practices of HCWs, levels of adherence of personnel to recommended handwashing practices, and factors adversely affecting adherence . New studies of the in vivo efficacy of alcohol-based hand rubs and the low incidence of dermatitis associated with their use are reviewed . Recent studies demonstrating the value of multidisciplinary hand-hygiene promotion programs and the potential role of alcohol-based hand rubs in improving hand-hygiene practices are summarized . Recommendations concerning related issues (e.g., the use of surgical hand antiseptics, hand lotions or creams, and wearing of artificial fingernails) are also included. Plant Cell, 2002 Nov, 14(11), 2837 - 47 Molecular mechanisms of proline-mediated tolerance to toxic heavy metals in transgenic microalgae; Siripornadulsil S et al.; Pro has been shown to play an important role in ameliorating environmental stress in plants and microorganisms, including heavy metal stress . Here, we describe the effects of the expression of a mothbean delta(1)-pyrroline-5-carboxylate synthetase (P5CS) gene in the green microalga Chlamydomonas reinhardtii . We show that transgenic algae expressing the mothbean P5CS gene have 80% higher free-Pro levels than wild-type cells, grow more rapidly in toxic Cd concentrations (100 microM), and bind fourfold more Cd than wild-type cells . In addition, Cd-K edge extended x-ray absorption fine structure studies indicated that Cd does not bind to free Pro in transgenic algae with increased Pro levels but is coordinated tetrahedrally by sulfur of phytochelatin . In contrast to P5CS-expressing cells, Cd is coordinated tetrahedrally by two oxygen and two sulfur atoms in wild-type cells . Measurements of reduced/oxidized GSH ratios and analyses of levels of malondialdehyde, a product of the free radical damage of lipids, indicate that free Pro levels are correlated with the GSH redox state and malondialdehyde levels in heavy metal-treated algae . These results suggest that the free Pro likely acts as an antioxidant in Cd-stressed cells . The resulting increased GSH levels facilitate increased phytochelatin synthesis and sequestration of Cd, because GSH-heavy metal adducts are the substrates for phytochelatin synthase. J Periodontol, 2002 Oct, 73(10), 1133 - 40 The role of CD4+ cells in vivo on the induction of the immune response to Porphyromonas gingivalis in mice; Sosroseno W et al.; BACKGROUND: It has previously been suggested that CD4+ T cells play a pivotal role in regulating the immune response to periodontal pathogens . The aim of the present study therefore was to determine delayed type hypersensitivity (DTH), spleen cell proliferation, serum and splenic anti-Porphyromonas gingivalis antibody levels, and lesion sizes following challenge with viable P . gingivalis in CD4-depleted BALB/c mice immunized with P . gingivalis outer membrane proteins (OMP) . METHODS: Four groups of BALB/c mice were used . Groups 1 and 2 were injected intraperitoneally (ip) with saline for 3 consecutive days and then weekly throughout the experiment . Groups 3 and 4 were injected ip with rat immunoglobulin and a monoclonal rat anti-mouse CD4 antibody, respectively . Two days later, group 1 mice were injected ip with saline only, while all the other groups were immunized ip with P gingivalis OMP weekly for 3 weeks . One week later following the last immunization of OMP, 3 separate experiments were conducted to determine: 1) the DTH response to P gingivalis OMP by measuring footpad swelling; 2) the levels of antibodies to P gingivalis in serum samples and spleen cell cultures using an enzyme-linked immunosorbent assay, as well as spleen cell proliferation after stimulation with OMP; and 3) the lesion sizes after a subcutaneous challenge with viable P . gingivalis cells . RESULTS: In CD4+ T-cell-depleted mice (group 4), the DTH response and antigen-stimulated cell proliferation were significantly suppressed when compared to groups 2 and 3 . Similarly, the levels of serum and splenic IgM, IgG, and all IgG subclass antibodies to P . gingivalis OMP were depressed . Delayed healing of P gingivalis-induced lesions was also observed in the CD4+ T-cell-depleted group . CONCLUSIONS: This study has shown that depletion of CD4+ T cells prior to immunization with P gingivalis OMP led to the suppression of both the humoral and cell-mediated immune response to this microorganism and that this was associated with delayed healing . These results suggest that the induction of the immune response to P . gingivalis is a CD4+ T-cell-dependent mechanism and that CD4+ T cells are important in the healing process. Eur J Clin Microbiol Infect Dis, 2002 Oct, 21(10), 755 - 7 Epub 2002 Oct 03. Oro-dental bacteria in various atherosclerotic arteries; Taylor-Robinson D et al.; Chlamydia pneumoniae DNA has been detected in at least 40% of all major arteries affected by atherosclerosis, but several other microorganisms have also been detected . In this study, diseased vessels were evaluated for the presence of the DNA of seven oro-dental bacteria and two nonoral bacteria . A polymerase chain reaction technique was employed using primer pairs based on 16S rRNA genes . Of 32 specimens tested, 10 (31.2%) were DNA positive: seven for Actinobacillus actinomycetemcomitans and three for Prevotella intermedia . The DNA was found in specimens from the aorta and the iliac, internal mammary and coronary arteries . Eleven (35.4%) of 31 specimens had been shown previously to be positive for Chlamydia pneumoniae DNA . A mixture of chlamydiae and oro-dental bacteria was found in three cases . These findings may have implications for antibiotic prophylaxis of coronary heart disease if directed solely at Chlamydia pneumoniae. Vet Microbiol, 2002 Dec 20, 90(1-4), 447 - 59 Diagnosis of brucellosis by serology; Nielsen K; Serological diagnosis of brucellosis began more than 100 years ago with a simple agglutination test . It was realized that this type of test was susceptible to false positive reactions resulting from, for instance, exposure to cross reacting microorganisms . It was also realized that this test format was inexpensive, simple and could be rapid, although results were subjectively scored . Therefore, a number of modifications were developed along with other types of tests . This served two purposes: one was to establish a rapid screening test with high sensitivity and perhaps less specificity and a confirmatory test, usually more complicated but also more specific, to be used on sera that reacted positively in screening tests . This led to another problem: if a panel of tests were performed and they did not all agree, which interpretation was correct? This problem was further compounded by the extensive use of a vaccine which gave rise to an antibody response similar to that resulting from field infection . This led to the development of an assay that could distinguish vaccinal antibody, starting with precipitin tests . These tests did not perform well, giving rise to the development of primary binding assays . These assays, including the competitive enzyme immunoassay and the fluorescence polarization assay are at the apex of current development, providing high sensitivity and specificity as well as speed and mobility in the case of the fluorescence polarization assay . J Theor Biol, 2002 Nov 21, 219(2), 137 - 52 Axisymmetric bioconvection in a cylinder; Ghorai S et al.; In three-dimensional bioconvection, the regions of rising and sinking fluid are dissimilar . This geometrical effect is studied for axisymmetric bioconvection in a cylindrical cell with stress-free (i.e . normal velocity and tangential stress vanish) lateral and top boundaries, and rigid bottom boundary . Using the continuum model of Pedley et al . (1988, J . Fluid Mech.195, 223-237) for bioconvection in a suspension of swimming, gyrotactic microorganisms, the structure and stability of an axisymmetric plume in a deep chamber are investigated . The system is governed by the Navier-Stokes equations for an incompressible fluid coupled with a microorganism conservation equation . These equations are solved numerically using a conservative finite-difference scheme . Comparisons are made with two-dimensional bioconvection. Mol Immunol, 2002 Nov, 39(7-8), 465 - 73 Evaluation and clinical interest of mannan binding lectin function in human plasma; Dumestre-Perard C et al.; The mannan binding lectin (MBL) plays a major role in innate immunity through its ability to activate complement upon binding to carbohydrate arrays on the surface of various microorganisms . The question of a possible association of the MBL structural gene polymorphism and the oligomeric state of MBL was poorly documented . For these reasons, it appears difficult to evaluate MBL in blood patients on the only basis of protein contents, even in combination with MBL genotyping . This study reports a method to calculate a specific activity for circulating MBL, that relies on: (i) the availability of purified MBL; and (ii) a simplified MBL activity assay based on complement activation . The three-step MBL purification from human plasma reported here is characterized by a highly purified MBL, that occurs in two different oligomeric forms . The results on the specific activity of these forms show that the higher oligomeric forms of MBL have the ability to induce C4 cleavage more efficiently than the corresponding lower oligomers . The usefulness of this approach is illustrated by its potential interest in the biological exploration of certain pathology, for example in the follow-up of chronic hepatitis C . Further investigation is needed to establish whether MBL specific activity (MBLsa) is correlated to the polymorphic state of the molecule . The relative simplicity of the test described here allows better investigation on the relationship between MBL biological activity and its genotype. Crit Rev Oncol Hematol, 2002 Nov, 44(2), 163 - 74 Fever and neutropenia in cancer patients: the diagnostic role of cytokines in risk assessment strategies; Oude Nijhuis CS et al.; Cancer patients treated with chemotherapy are susceptible to bacterial infections . Therefore, all neutropenic cancer patients with fever receive standard therapy consisting of broad-spectrum antibiotics and hospitalization . However, febrile neutropenia in cancer patients is often due to other causes than bacterial infections . Therefore, standard therapy should be re-evaluated and new treatment strategies for patients with variable risk for bacterial infection should be considered . This paper reviews the changing spectrum of microorganisms and resistance of microorganisms to antibiotics in infection during neutropenia and discusses new strategies for the selection of patients with low-risk for bacterial infection using clinical and biochemical parameters such as acute phase proteins and cytokines . These low-risk patients may be treated with alternative therapies such as oral antibiotics, early discharge from the hospital or outpatient treatment. Ann Pathol, 2002 Sep, 22(4), 289 - 304 {Cytopathologist's role in detecting and identifying pathogens}; Hofman P et al.; This review highlights the contribution of the cytopathologist in the diagnosis of infectious diseases, with emphasis on the detection and identification of microorganisms in various cytologic specimens . Morphological analysis of the main bacteria, parasites and mycoses observed in cytologic specimens, as well as the cytopathogenic effects of the viruses are discussed . The differential diagnosis (contaminants and exogenous or endogenous foreign bodies) is discussed . Finally, the main pathogens noted in each specimen are briefly reviewed. Dev Cell, 2002 Oct, 3(4), 581 - 92 An immune-responsive Serpin regulates the melanization cascade in Drosophila; De Gregorio E et al.; In arthropods, the melanization reaction is associated with multiple host defense mechanisms leading to the sequestration and killing of invading microorganisms . Arthropod melanization is controlled by a cascade of serine proteases that ultimately activates the enzyme prophenoloxidase (PPO), which, in turn, catalyzes the synthesis of melanin . Here we report the biochemical and genetic characterization of a Drosophila serine protease inhibitor protein, Serpin-27A, which regulates the melanization cascade through the specific inhibition of the terminal protease prophenoloxidase-activating enzyme . Our data demonstrate that Serpin-27A is required to restrict the phenoloxidase activity to the site of injury or infection, preventing the insect from excessive melanization. Dig Liver Dis, 2002 Sep, 34 Suppl 2, S8 - 11 Probiotics: clinics and/or nutrition; Morelli L; The primary claim of probiotics is their beneficial influence on the intestinal ecosystem, which, in turn, may provide protection against gastro-intestinal infections . The positive actions on human health include antagonistic activity against pathogens, anti-allergic effects and other effects on the immune system . In recent years, the concept of probiotics evolved from food with beneficial, even if not clearly established, effects on well-being, to the new area of the nutraceuticals or functional foods . This concept was developed further into the biotherapeutics, which are the clinical applications of probiotic microorganisms originally selected for food use . Whereas several of these health claims remain controversial, well-planned clinical trials increasingly support the claims for some carefully selected probiotic strains . Aim of this review is to provide a first attempt to evaluate these two areas of use of probiotics and to draw a borderline between them . Only a more in-depth knowledge of the mechanisms underlying the probiotic effects will allow a clear discrimination between the food use and the clinical use. Methods Mol Med, 2003, 75, 121 - 33 Myeloperoxidase promoter region polymorphism and lung cancer risk; Wu X et al.; Historically, myeloperoxidase activity and subsequent production of hypochlorous acid has been associated with the killing of host-invading microorganisms (bacteria, viruses, and fungi) . Currently, there is a wealth of evidence that the MPO polymorphism and enzyme activity is associated with a wide range of pathological and biological processes, including lung cancer carcinogenesis . Although the molecular epidemiology reports reviewed in this chapter are not in complete agreement on all aspects of their findings, it is evident that the MPO polymorphism contributes to the modulation of overall lung cancer risk . Four of the five molecular epidemiologic studies reviewed in this chapter utilized similar case-control study designs with quite different sources of populations . These studies all demonstrate that the MPO variant genotype modulates overall lung cancer risk . However, these studies are not in agreement regarding age and gender effects, and gene-environmental interactions . The nested case control study designed utilized by Misra et al . (35) is a valid and sound approach, but their results found no evidence of an association . Certainly, heterogeneity in study populations can contribute to the variability between these studies . Additionally, epidemiologic issues such as case-control matching and sources of control populations may contribute to the conflicting findings . Thus, the publication of inconsistent or null studies as well as other positive findings is certainly encouraged to elucidate the range of effects associated with the MPO polymorphism and lung cancer risk. J Pediatr Surg, 2002 Nov, 37(11), 1540 - 2 Does colonization of Helicobacter pylori in the heterotopic gastric mucosa play a role in bleeding of Meckel's diverticulum? Ergun O, Celik A, Akarca US, Sen T, Alkanat M, Erdener A. BACKGROUND/PURPOSE: Helicobacter pylori is a microorganism known to colonize in gastric type of mucosa and is associated with gastritis and peptic ulceration . The aim of the study was to determine whether colonization of H pylori in heterotopic gastric mucosa plays a role in bleeding of Meckel's diverticulum . METHODS: Histopathologic slides of patients who had undergone resection of Meckel's diverticulum in recent 5 years were reexamined for the presence of H pylori in heterotopic gastric mucosa . Polimerase chain reaction (PCR) test was used to trace the genetic material of urease gene and 16s rDNA amplifications for H pylori . RESULTS: Thirteen of the 30 histopathologic slides of Meckel's diverticula had heterotopic gastric mucosa . Ten of the 13 patients presented with acute bleeding of the diverticula, whereas 3 of them were asymptomatic . None of the 13 gastric mucosa bearing diverticula were colonized with H pylori . PCR was unable to show any trace of genetic material for H pylori . CONCLUSION: Although the role of H pylori is well established in the gastric mucosal ulceration, its presence is not essentially required to induce "heterotopic gastritis" that may result in bleeding of the Meckel's diverticulum . . Rev Neurol (Paris), 2002 Oct, 158(10 Pt 1), 993 - 7 {Tick-borne neurological diseases}; Christmann D et al.; Some microorganisms transmitted to man via tick bite are responsible for infections, which can be associated with neuro-meningeal complications . TBE virus is the most frequent virus associated with potentially severe neurological lesions . No treatment is available so far . The most frequent bacterial diseases in which neurological complications may appear are Lyme borreliosis, Q fever and some rickettsial infections . More recently ehrlichiosis have emerged as new infections that may be associated with neuro-meningeal complications . Appropriate antibiotic therapy may be used for the treatment of these bacterial infections, but no vaccine is available so far. Best Pract Res Clin Rheumatol, 2002 Sep, 16(4), 507 - 22 Reactive arthritis or post-infective arthritis? Keat A. Infective mechanisms probably underlie a wide range of inflammatory arthropathies . There appears to be a spectrum of mechanisms ranging from the frankly septic, through low-grade infection with very small numbers of microorganisms in the joint to arthritides in which no hard evidence for an infective cause exists . In the midst of the spectrum lie 'post-infective' and 'reactive' arthritides, characterized clinically, genetically and by epidemiological links with infection . Identification of bacterial components within joint material from such patients suggested that the causes of the arthritis had been found . It is now clear that many bacteria are present in inflamed joints; establishing their significance will be of crucial importance, but not easy. Med Clin (Barc), 2002 Oct 19, 119(13), 481 - 4 {Community-acquired pneumonia: serum adenosine-deaminase activity in the aetiological diagnosis}; Fernandez Alvarez R et al.; BACKGROUND: Adenosine deaminase (ADA) is a cytoplasmic enzyme which activity is increased in disorders that stimulate cells involved in the immune system . In community-acquired pneumonia (CAP), increased levels of serum ADA have been associated with the presence of atypical microorganisms as the source of the former . Previous studies have shown ADA increases in non-infectious diseases . We evaluated the factors that may influence plasmatic ADA (ADAp) levels in CAP patients . PATIENTS AND METHODS: A study with cases (245 episodes of CAP) and controls (49) was designed, and the differences in ADAp activity with regard to organisms, comorbidity factors and complications were analyzed . A logistic regression analysis was performed . RESULTS: CAP caused by atypical microorganisms were found to have increased ADAp values . Variables that independently increased ADAp levels were: atypical etiology (OR = 5.9), liver disease (OR = 5.8), diabetes mellitus (OR = 1.9), and prior antibiotic consumption (OR = 1.7) . CONCLUSIONS: ADAp is an etiologic marker that could be useful in the empiric approach of the treatment of CAP. Crit Rev Biotechnol, 2002, 22(3), 245 - 79 Botryococcus braunii: a renewable source of hydrocarbons and other chemicals; Banerjee A et al.; Botryococcus braunii, a green colonial microalga, is an unusually rich renewable source of hydrocarbons and other chemicals . Hydrocarbons can constitute up to 75% of the dry mass of B . braunii . This review details the various facets of biotechnology of B . braunii, including its microbiology and physiology; production of hydrocarbons and other compounds by the alga; methods of culture; downstream recovery and processing of algal hydrocarbons; and cloning of the algal genes into other microorganisms . B . braunii converts simple inorganic compounds and sunlight to potential hydrocarbon fuels and feedstocks for the chemical industry . Microorganisms such as B . braunii can, in the long run, reduce our dependence on fossil fuels and because of this B . braunii continues to attract much attention. Presse Med, 2002 Oct 5, 31(32), 1512 - 6 {The risk of contagious epidemics in geriatric facilities}; Trivalle C; THE RISKS OF EPIDEMICS IN INSTITUTIONAL SETTINGS: An epidemic must be suspected when an increase in the number of cases of a same type of infection is observed . Numerous microorganisms are responsible for epidemics in geriatric facilities: viruses, bacteria and parasites . In the case of an epidemic, a certain number of specific measures must be taken in order to prevent the transmission of infection and eradicate the epidemic . IN THE CASE OF INFLUENZA: Other than the vaccination of elderly institutional residents, that of the nursing staff appears essential . If a severe epidemic occurs, specific antivirals can be used, three of which are already available . IN THE CASE OF PNEUMOCOCCI: Examples of epidemics of pneumococcal infections in elderly institutional residents in the United States underlines the interest of pneumococcal vaccines, particularly since the strain responsible corresponded to a serotype contained in the 23 valence vaccine . WITH REGARD TO SCABIES: All the patients and all the staff must be treated on the same day and at the same time their clothing and bed linen . All persons in contact with the patient, the families and friends of the staff, their clothes and the environement must be treated. Ann Ital Med Int, 2002 Jul-Sep, 17(3), 157 - 65 {Probiotics: history, definition, requirements and possible therapeutic applications}; Montalto M et al.; The ingestion of probiotics is associated with various beneficial effects on human health and modifies the physiological homeostasis of the intestinal flora . Probiotics are microorganisms with some particular characteristics: human origin, safety in human use, bile and acid resistance, survival in the intestine, at least temporary colonization of the human gut, adhesion to the mucosa and bacteriocine production . Thanks to these characteristics, probiotics block the invasion of human intestinal cells by the enteroinvasive bacteria . Furthermore, they should be able to stimulate and modulate the intestinal immune response, and to protect and stabilize the mucosal barrier . Finally, the efficacy of probiotics should be evident and documented with valid studies . All their properties should be maintained during processing and storage . Probiotics are usually used to protect the host from pathogens . With regard to this, they are useful in the prevention of antibiotic and traveler's diarrhea and they may play a role in the management of gastric Helicobacter pylori infection . Furthermore, their efficacy in the treatment of infectious diarrhea, in inflammatory bowel diseases, in pouchitis and in food allergy has been shown . Probiotics can improve the symptoms of irritable bowel syndrome and of lactose malabsorption . Finally, it has been suggested that such microorganisms may play a role in the prevention of carcinogenesis and of tumor growth. Adv Perit Dial, 2002, 18, 177 - 83 Peritonitis in continuous ambulatory peritoneal dialysis: cytokines in peritoneal fluid and blood; Enriquez J et al.; Cytokines are soluble mediators of the immune system that regulate the response to antigens and microorganisms . In patients on continuous ambulatory peritoneal dialysis (CAPD) who have peritonitis, an inflammatory process exists that must be understood if susceptibility to, and the mechanisms of, complications such as fibrosis and others are to be understood . To that end, we studied 9 CAPD patients with peritonitis . The case series was conducted in Popayan, Colombia, at the RTS Cauca dialysis unit and the University of Cauca hospital, a tertiary health care facility . Peritonitis was diagnosed by standard clinical and laboratory criteria . Using flow cytometry, we measured the percentage production of intracellular cytokines {interleukin-1 alpha (IL-1 alpha), IL-6, IL-12, tumor necrosis factor alpha (TNF alpha), IL-4, and interferon-gamma (IFN-gamma) in T lymphocytes from blood and peritoneal fluid . Among the studied patients, all (100%) produced high levels of IL-1, IL-6, TNF alpha, IL-12, and IL-4 in both fluids (blood: 89% +/- 63% of cells; peritoneal fluid: 81.6% +/- 10.1% of cells) . In blood, 25% of patients produced IFN-gamma (mean: 15.7% of cells), showing that 75% of patients had the TH2 pattern, and 25% were close to TH0 . In peritoneal fluid, 34% of patients produced IFN-gamma spontaneously (mean: 24.5% of cells), indicating that 66% of patients were TH2, and 34% were close to TH0 . After stimulation, expression of cytokines, including IFN-gamma (39% of T cells), was increased, and high production of IL-4 indicated that 25% of patients were TH2, and 75% were TH0 . In peritoneal fluid, production of cytokines, including IFN-gamma, was increased, with high production of IL-4, indicating switching from TH2 (34% of patients) to TH0 (66% of patients) . Of the studied patients, 35% had a CD4:CD8 ratio < 1.1 in blood, and also produced IL-12 (94.5% of cells) and IFN-gamma (30% of cells), as compared with patients in whom the CD4:CD8 ratio was > 1.2 . Patients on CAPD who have peritonitis produce large amounts of pro-inflammatory and TH2 cytokines . More IFN-gamma is produced in peritoneal fluid than in blood, which suggests more inflammation . Immunodeviation TH2 is seen in blood and peritoneal fluid of CAPD patients with peritonitis . Patients with a CD4:CD8 ratio < 1.1 produce more IFN-gamma and IL-12, and are more able to switch from TH2 to TH0. Arkh Patol, 2002 Jul-Aug, 64(4), 37 - 40 {Clinico-morphological and molecular-genetic correlations in the stomach pathology associated with Helicobacter pylori}; Chukov SZ et al.; Some clinicomorphological correlations are made in Helicobacter pylori (H . pylori) associated diseases of the gastroduodenal zone . Genes vacA and cagA in the H . pylori genome were revealed by PCR in stomach biopsies of infected patients and the results were compared with histological changes in the patients' mucous membrane . Specific IgG antibodies to some H . pylori antigens were detected in the serum and a comparative analysis of serological and histological data was performed . The findings suggest that H . pylori associated gastroduodenal pathology is determined by interaction between macro- and microorganisms taking account of genetic characteristics of the infective microorganism, its phenotypic variations and properties of inflammatory and immune response of the host. Acta Otorrinolaringol Esp, 2002 Jun-Jul, 53(6), 427 - 30 {Subdural empyema of sinus origin caused by Gemella morbillorum, a strange etiology}; Martinez Beneito MP et al.; We present a case of frontal sinusitis complicated with a subdural empyema, in which the identified microorganism was Gemella morbillorum, a frequent host of the aerodigestive tract and occasionally related to infections . The problem was resolved successfully using endoscopic surgery and an external approach of the sinus . Afterwards it was completed with a subdural drainage through craniotomy . Subdural empyema is a rare complication of sinusitis although very severe . We want to emphasize the importance of early diagnosis of intracranial complications, the need of a detailed microbiology test the method used to obtain samples, and the convenience of a combined approach by the otolaryngologists and the neurosurgeons for its complete drainage. Nippon Ishinkin Gakkai Zasshi, 2002, 43(4), 203 - 13 A new experimental murine aspergillosis model to identify strains of Aspergillus fumigatus with reduced virulence; Sarfati J et al.; Experimental animals are an obligate screen to investigate microorganism pathogenicity . Numerous animal models have been used to analyse the virulence of the opportunistic human pathogen Aspergillus fumigatus but none of the experimental models used previously have been satisfactory . This report discuss these models and presents a murine model of pulmonary aspergillosis that is very easy and the most adapted to compare the pathogenicity of A . fumigatus strains . Strains to be tested are inoculated intranasally and synchronously to mice and strains isolated from the lung of mice killed by the infection are typed . The number of colonies recovered is directly correlated to the virulence of the strain. J Environ Monit, 2002 Oct, 4(5), 667 - 72 Volatile metabolites from microorganisms grown on humid building materials and synthetic media; Claeson AS et al.; Growth of different microorganisms is often related to dampness in buildings . Both fungi and bacteria produce complicated mixtures of volatile organic compounds that include hydrocarbons, alcohols, ketones, sulfur- and nitrogen-containing compounds etc . Microbially produced substances are one possible explanation of odour problems and negative health effects in buildings affected by microbial growth . A mixture of five fungi, Aspergillus versicolor, Fusarium culmorum, Penicillium chrysogenum, Ulocladium botrytis and Wallemia sebi were grown on three different humid building materials (pinewood, particle board and gypsum board) and on one synthetic medium . Six different sampling methods were used, to be able to collect both non-reactive volatile organic compounds and reactive compounds such as volatile amines, aldehydes and carboxylic acids . Analysis was performed using gas chromatography, high-performance liquid chromatography and ion chromatography, mass spectrometry was used for identification of compounds . The main microbially produced metabolites found on pinewood were ketones (e.g . 2-heptanone) and alcohols (e.g . 2-methyl-1-propanol) . Some of these compounds were also found on particle board, gypsum board and the synthetic medium, but there were more differences than similarities between the materials . For example, dimethoxymethane and 1,3,5-trioxepane and some nitrogen containing compounds were found only on particle board . The metabolite production on gypsum board was very low, although some terpenes (e.g . 3-carene) could be identified as fungal metabolites . On all materials, except gypsum board, the emission of aldehydes decreased during microbial growth . No low molecular weight carboxylic acids were identified. Biotechniques . 2002 Oct;33(4):798, 800, 802 passim. Microplate diffusion assay for screening of beta-glucanase-producing microorganisms; Zantinge JL et al.; A method is described to screen fungal strains rapidly for overexpression of extracellular beta-1,4-endoglucanase in the presence of high levels of sugar compounds . The semi-quantitative assay utilizes microplates in a 96-well format and an azurine dye covalently cross-linked (AZCL) chromogenic substrate . The digestion of AZCL-hydroxyethyl-beta-1,4-endoglucanase results in the release of a blue dye directly proportional to the amount of enzyme activity present in the sample . Sample absorbance was read at 590 nm . and the enzyme activity was determined by reference to a standard curve . The results from the microplate diffusion assay were similar to the results derived from the Ostazin Brilliant Red-hydroxyethyl cellulose assay . The technique described allowed the rapid comparison and screening beta-1,4-glucanase activity directly in spent fungal supernatant, from cultures grown in potato dextrose broth . The method could also be easily adapted for the screening of the presence of other activities such as beta-1,3-glucanase activity by using either AZCL-beta-glucan or AZCL-pachyman in place of the AZCL-hydroxyethyl-cellulose . This assay could be used to measure supernatant within an activity range of 0.1-2 U/mL Hepatogastroenterology, 2002 Nov-Dec, 49(48), 1484 - 6 Infectious factors in acute acalculous cholecystitis; Teixeira JP et al.; BACKGROUND/AIMS: The infection is shown as a secondary phenomenon in lithiasic acute cholecystitis . However, its importance in acute gallbladder inflammation without lithiasis has not been sufficiently clarified . In this context, we tried to investigate this issue by resorting to animal research . Similarly we studied the repercussions of Diclofenac (NSAID) in bacteriological features of the bile in the operated animals . METHODOLOGY: We provoked acute cholecystitis in pigs by the ligature of biliary pedicle associated to the gallbladder introduction of "Lysophosphatidylcoline" . Two series of animals were used: P1 (pigs not treated with Diclofenac) with 17 pigs and P2 (pigs treated with Diclofenac) with 23 . In the latter group we associated the administration of Diclofenac given before the production of acute cholecystitis and administered for 4 days . The microbiology of the bile was studied in 34 pigs . The following parameters were valorized: histological gallbladder lesions and microorganisms in bacteriological examination of the bile . RESULTS: Severe lesions of acute cholecystitis were registered in 82.3% of the animals . Positive bile cultures were registered in 64.1% of group P1 and 65.5% in the group P2 . The simultaneous administration of Diclofenac in the P2 group did not cause a regression of gallbladder lesions or in microbial agents in the bile in relation to the first group (P = 0.649, and P = 0.781, respectively) . CONCLUSIONS: Our investigations allow us to conclude by valorization the infection factor in acute acalculous cholecystitis . On the other hand, the absence of repercussion of Diclofenac in acute acalculous cholecystitis was demonstrated not only in histological gallbladder lesions, but also in bile infection. Immunobiology, 2002 Sep, 205(4-5), 498 - 517 Lung surfactant protein D (SP-D) and the molecular diverted descendants: conglutinin, CL-43 and CL-46; Hansen S et al.; Lung surfactant protein D (SP-D), conglutinin, CL-43 and CL-46 belong to a group of proteins designated collectins that, besides a common structure made of a collagen-like region and a C-type lectin domain, are important components of the innate immune defence . They all bind complex glycoconjugates on microorganisms thereby inhibiting infection, enhancing the clearance by phagocytes and modulating the immune response . In addition, SP-D inhibits the generation of radical oxygen species or the propagation of lipid peroxidation . Knock-out mice deficient in SP-D have a disturbed homeostasis of pulmonary surfactant and suffer from oxidative stress leading to pulmonary inflammation upon microbial challenge . Conglutinin, CL-43 and CL-46 have in contrast to the rest of the collectin family only been found in cattle . During the characterization of the genes encoding conglutinin, CL-43 and CL-46 we observed several features showing that the additional bovine collectins are diverted molecular descendants of an ancestral SP-D gene . Since structural similarity often associates with common functionality, some of SP-D's effector mechanisms may apply to conglutinin, CL-43 and CL-46--and vice versa . This review focus on the structural and functional relationship of this group of collectins. Immunobiology, 2002 Sep, 205(4-5), 467 - 75 The biological functions of MBL-associated serine proteases (MASPs); Hajela K et al.; The Mannose-binding lectin-associated serine proteases (MASPs) have been the subject of intensive research particularly over the past 10 years . First one, then two, and currently 3 MASPs have been characterized . Initially it was thought likely that the MBL + MASPs system would resemble very closely the C1 complex of the complement classical pathway, and that MASP1 and MASP2 would have similar activities to their classical pathway homologues C1r and C1s . MASP2 does certainly have similar activities to C1s, but MASP1 does not have the activities of either C1r or C1s . MASP1 has been thought to act on the complement system by cleaving C3 directly, but work with recombinant and purified native MASP1 shows that direct C3 cleavage by this protease is very slow, and may not be biologically significant . MASP1 and MASP2 appear not to have such a narrow specificity as C1r and C1s, and may have significant substrates other than complement proteins . As an example, MASP1 does cleave fibrinogen, releasing fibrinopeptide B (a chemotactic factor) and also cleaves and activates plasma transglutaminase (Factor XIII) . These reactions are also relevant to defence against microorganisms, and may represent a biologically significant action of MASP1. Immunobiology, 2002 Sep, 205(4-5), 433 - 45 Structural and functional aspects of complement activation by mannose-binding protein; Wallis R; Serum mannose-binding protein (MBP) is the first component of the lectin pathway of the complement cascade . It binds to sugars on the surface of pathogenic microorganisms and triggers complement fixation by activating an associated serine protease, designated MBP-associated serine protease-2 (MASP-2) . Recent studies have provided insight into the interactions between MBP and MASP-2 that trigger complement activation . MBP/MASP complexes share many features with the C1 complex of the classical pathway . The relatively simple MBP/MASP complexes serve as useful models for understanding activation of the classical pathway of the complement cascade. CLAO J, 2002 Oct, 28(4), 177 - 80 Spherical indentations of human and rabbit corneal epithelium following extended contact lens wear; Ladage PM et al.; PURPOSE: Mucin balls appear to cause spherical indentations in the corneal epithelium during silicone hydrogel extended contact lens wear . The purpose of this report is to describe and quantify these spherical indentations, as examined in the human cornea by in vivo confocal microscopy and by in vitro immunocytochemistry in the rabbit cornea . METHODS: Confocal images of full-thickness corneal epithelium were taken from three human patients participating in a 1-year extended contact lens-wear trial . Diameter and depth of the indentations were determined and measured . Two rabbit corneas showing identical indentations were stained with propidium iodide (nuclear stain) and Ki-67 (proliferation marker) and were examined using a laser scanning confocal microscope . RESULTS: The diameter of the spherical indentations is largest on the epithelial surface, ranging from 33.9 to 78.8 microm . Indentations form spherical sections whose depth variably extends into the corneal epithelium, reaching as far as the basal lamina . The rabbit model showed no epithelial nuclei within the indentation . Furthermore, stromal cells localized immediately beneath the indentations were positive for Ki-67 (proliferation) . DISCUSSION: Spherical indentations of the corneal epithelium induced by mucin balls appear to be gaps or holes that can extend deep into the corneal epithelium . Indentations may potentially open a pathway for infectious microorganisms to penetrate the cornea . Surprisingly, stromal cells immediately beneath the holes were stimulated to proliferate, and there seemed to be an increase in localized cell density. Curr Opin Hematol, 2002 Nov, 9(6), 485 - 9 Cellular immunity to viral and fungal antigens after stem cell transplantation; Einsele H et al.; Cellular immunity to viral and fungal antigens is often suppressed after stem cell transplantation . Viral infections, in particular infections with cytomegalovirus and Epstein-Barr virus, are often reactivated after allogeneic stem cell transplantation . Pathogenetic factors are immune deficiency and T-cell stimulation in the course of graft-versus-host reactions . Fungal infections are opportunistic infections derived with ubiquitous microorganisms . Immune deficiency, neutropenia, steroid treatment, and antibiotic treatment contribute to the pathogenesis . The study of viral and fungal immunity after allogeneic stem cell transplantation gives insight into the reconstitution of the immune system and tolerance. J Biol Chem, 2003 Jan 17, 278(3), 1735 - 43 Epub 2002 Oct 21. Biochemical and structural studies of malate synthase from Mycobacterium tuberculosis; Smith CV et al.; Establishment or maintenance of a persistent infection by Mycobacterium tuberculosis requires the glyoxylate pathway . This is a bypass of the tricarboxylic acid cycle in which isocitrate lyase and malate synthase (GlcB) catalyze the net incorporation of carbon during growth of microorganisms on acetate or fatty acids as the primary carbon source . The glcB gene from M . tuberculosis, which encodes malate synthase, was cloned, and GlcB was expressed in Escherichia coli . The influence of media conditions on expression in M . tuberculosis indicated that this enzyme is regulated differentially to isocitrate lyase . Purified GlcB had K(m) values of 57 and 30 microm for its substrates glyoxylate and acetyl coenzyme A, respectively, and was inhibited by bromopyruvate, oxalate, and phosphoenolpyruvate . The GlcB structure was solved to 2.1-A resolution in the presence of glyoxylate and magnesium . We also report the structure of GlcB in complex with the products of the reaction, coenzyme A and malate, solved to 2.7-A resolution . Coenzyme A binds in a bent conformation, and the details of its interactions are described, together with implications on the enzyme mechanism. Blood, 2003 Jan 15, 101(2), 711 - 21 Epub 2002 Sep 05. Glycosphingolipid expression in acute nonlymphocytic leukemia: common expression of shiga toxin and parvovirus B19 receptors on early myeloblasts; Cooling LL et al.; Glycosphingolipids (GSLs) are complex macromolecules on cell membranes that have been shown to play a role in neutrophil differentiation, activation, phagocytosis, and adhesion to both microorganisms and vascular endothelium . Because GSLs are often cryptic antigens on cell membranes, little is known regarding GSL expression in early myelopoiesis . To study the latter, myeloblasts were collected from patients with acute nonlymphocytic leukemia (ANLL) who required therapeutic leukocytopheresis for hyperleukocytosis . The neutral GSLs were isolated and identified by high-performance thin-layer chromatography (HPTLC), HPTLC immunostaining, gas chromatography, nuclear magnetic resonance, and fast atom bombardment-mass spectrometry . Like mature peripheral blood neutrophils, myeloblasts expressed glucosylceramide, lactosylceramide, and the neolacto-family GSLs, lactotriaosylceramide and neolactotetraosylceramide . Unlike neutrophils and chronic myeloid leukemia, most ANLL samples also expressed the globo-series GSLs, globotriaosylceramide and globotetraosylceramide . Globo GSL expression was strongly associated with a myeloblastic (ANLL M0-M2) and monoblastic phenotype (M5) . A weak association was also noted with expression of either lymphoid (P <.10) or early hematopoietic markers (terminal deoxynucleotidyl transferase {TdT}, CD34; P <.10) . Globo-positive ANLL samples bound both shiga toxin and parvovirus B19 on HPTLC immunostaining . Based on these findings, we propose that neolacto and globo GSLs are expressed during early myeloid differentiation . Globotriaosylceramide expression on myeloblasts, and possibly myeloid stem cells, may have important implications for the use of shiga toxin as an ex vivo purging agent in autologous stem cell transplantation . Expression of globotetraosylceramide, the parvovirus B19 receptor, on myeloblasts may also explain the association between B19 infection, aplastic anemia, and chronic neutropenia of childhood. Blood, 2002 Dec 1, 100(12), 3908 - 18 Epub 2002 Jul 25. Host defense and inflammatory gene polymorphisms are associated with outcomes after HLA-identical sibling bone marrow transplantation; Rocha V et al.; We made the hypothesis that donor and recipient gene polymorphisms that drive the host response to microorganisms could be associated with infections after bone marrow transplantation (BMT) . HLA-identical BMT was performed for patients with acute (n = 39) or chronic leukemia (n = 68) . Genotyping was performed in 107 D/R DNA pairs for gene polymorphisms of cytokines (tumor necrosis factor-alpha {TNF-alpha} and TNF-beta, interleukin-1 receptor antagonist {IL-1Ra}, IL-6, and IL-10), adhesion molecules (CD31 and CD54), Fcgammareceptors (FcgammaRIIa, IIIa, IIIb), mannose-binding lectin (MBL), and myeloperoxidase (MPO) . First infection (overall) and first episodes of bacterial, viral, or invasive fungal infection were studied retrospectively for 180 days after BMT . Univariate and multivariate analyses, using death as a competing event, were performed to study risk factors . In multivariate analysis, first overall infections were increased in patients with the FcgammaRIIa R-131 genotype (hazard ratio {HR} = 1.92; P =.04), and severe bacterial infections were increased when the MPO donor genotype was AG or AA (HR = 2.16; P =.03) . Viral and invasive fungal infections were not influenced by any genetic factor studied . Interestingly, we also found that (1) time to neutrophil recovery was shorter when donors were FcgammaRIIIb HNA-1a/HNA-1b (HR = 1.77; P =.002); (2) donor IL-1Ra (absence of IL-1RN*2) increased the risk for acute graft-versus-host disease (GVHD) (II-IV) (HR = 2.17; P =.017); and (3) recipient IL-10 (GG) and IL-1Ra genotypes increased the risk for chronic GVHD (P =.03 and P =.03, respectively) . Finally, 180-day transplantation-related mortality rates were increased when donors were FcgammaRIIIb HNA-1a/HNA-1a or HNA-1b/HNA-1b (HR = 2.57; P =.05) and donor MPO genotype was AA (HR = 5.14; P =.004) . In conclusion, donor and recipient gene polymorphisms are informative genetic risk factors for selecting donor/recipient pairs and could help in the understanding of mechanisms involved in host defenses of BM transplant recipients. Blood, 2003 Jan 1, 101(1), 210 - 5 Epub 2002 Aug 15. Early macrophage influx to sites of cutaneous granuloma formation is dependent on MIP-1alpha /beta released from neutrophils recruited by mast cell-derived TNFalpha; von Stebut E et al.; Macrophages (MPhi) play a crucial role in the development of cutaneous granulomas (CGs) initiated by foreign bodies or invasive microorganisms . However, little is known about how MPhi are recruited to sites of CG formation . To test whether mast cells (MCs) contribute to early MPhi recruitment to developing granulomas, CGs were induced in MC-deficient Kit(W)/Kit(W-v) mice by injection of polyacrylamide gel (PAG) . Kit(W)/Kit(W-v) mice as well as mice deficient in the MC product TNFalpha exhibited markedly reduced MPhi numbers in CGs . MPhi recruitment was restored in Kit(W)/Kit(W-v) mice reconstituted with MCs from Kit(+/+) or TNFalpha(+/+), but not from TNFalpha(-/-) mice . MC-TNFalpha-dependent MPhi influx required prior recruitment of MIP-1alpha/beta-producing neutrophils (PMNs), as PMN depletion before induction of CGs completely inhibited MPhi influx, which was restored after reconstitution with PMN supernatants . These findings indicate that MPhi recruitment to cutaneous PAG- induced granulomas is the result of a sequence of inflammatory processes initiated by MC-derived TNFalpha followed by PMN influx and MIP-1a/beta release. J Appl Microbiol, 2002, 93(5), 800 - 9 Hygiene measures considering actual distributions of microorganisms in Japanese households; Ojima M et al.; AIMS: Effective household hygiene measures require that sources of bacterial contamination and the places to which contamination spreads be carefully identified . Therefore, a study was performed to examine the distribution of microorganisms throughout ordinary households in Japan, which has its own unique customs of daily life and food preparation . METHODS AND RESULTS: Using the stamping method, samples were taken from 100 different places and items in each of 86 households . This study found kitchens/dining rooms to have the greatest level of microbial contamination and bathrooms, the next highest level . Toilets (water closets) were found to have an unexpectedly low level of bacterial contamination . The largest bacterial counts were found on items such as drain traps, dish-washing sponges, counter towels, sinks, dish-washing tubs, and bathroom sponges . CONCLUSIONS: It is necessary to carefully identify both the items that can become instruments for spreading bacterial contamination and the places that easily become subject to secondary contamination, and then to take timely and effective disinfection/sanitizing measures . SIGNIFICANCE AND IMPACT OF THE STUDY: The data gathered in this study will be very valuable for anticipating the pathways over which bacteria are transported and prioritizing disinfection targets, to make effective disinfection possible. J Dermatol, 2002 Sep, 29(9), 547 - 55 Increased microorganisms DNA levels in peripheral blood monocytes from psoriatic patients using PCR with universal ribosomal RNA primers; Okubo Y et al.; It has long been suspected that systemic and focal infections cause or exacerbate psoriatic lesions . We previously showed that peripheral blood monocytes in psoriatic patients are activated and overproduce inflammatory cytokines . In addition, it has been reported that macrophages activated by ingesting microorganisms release tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta . Therefore we hypothesized that the monocytes in psoriatic patients may be activated by ingesting microorganisms and overproduce inflammatory cytokines . We examined the detection of microorganism DNA in monocytes from 15 patients with psoriasis vulgaris and from 12 healthy controls . DNA was extracted from monocytes, and a polymerase chain reaction (PCR) assay was performed for the detection using universal primers from conserved regions of the bacterial 16S ribosomal RNA gene or the fungal 18S rRNA gene . At the same time, we calculated the psoriasis area and severity index (PASI) scores and analyzed their correlations with the microorganisms DNA levels . The results showed that bacterial 16S DNA levels in monocytes were significantly higher in psoriatic patients than in controls . The fungal 18S DNA levels were also higher in psoriatic patients than in controls, but the differences were not significant . Although the microorganisms DNA levels in monocytes of psoriatic patients were high, there was no correlation between the bacterial DNA levels in monocytes of the psoriatics and PASI scores . Our study suggests that monocytes in psoriatic patients engulf more bacteria than there in controls, causing an activation of monocytes and triggering the formation of new lesions in the initial stages of psoriasis. Prikl Biokhim Mikrobiol, 2002 Sep-Oct, 38(5), 513 - 7 {Selection of microorganisms capable of degrading petroleum and petroleum products at decreased temperatures}; Belousova NI et al.; Of 150 cultures capable of degrading petroleum at +6 degrees C, 40 strains growing in the liquid mineral nutrient medium containing petroleum (2%) as a sole source of carbon were selected . Of them, 13 cultures displaying a petroleum degradation rate exceeding 25% were selected . Abilities of these cultures and their associations to utilize fuel oil and its components--oils and benzene resins--were studied . The culture exhibiting degradation rates of fuel oil, its oils, benzene resins, and petroleum amounting to 17, 26, 10, and 51%, respectively, was selected . This culture can be used for cleanup of petroleum pollution under cold climatic conditions. Salud Publica Mex, 2002 Sep-Oct, 44(5), 464 - 75 {Principal characteristics and diagnosis of the pathogenic groups of Escherichia coli}; Rodriguez-Angeles G; Escherichia coli colonizes the human intestinal tract within hours of birth and is considered a non-pathogenic member of the normal intestinal flora . However, there are six pathogenic groups that may produce diarrhea: enterotoxigenic (ETEC), enterohemorrhagic (EHEC), enteroinvasive (EIEC), enteropathogenic (EPEC), enteroaggregative (EAEC) and diffusely adherent (DAEC) groups . E . coli can be isolated and classified using traditional methods, by identifying its biochemical or serum characteristics . The pathogenic mechanisms may be studied in cell cultures and animal model assays, as well as more up to date molecular biology methods for study and diagnosis . The latter have proven that genes are involved in pathogenesis . The objective of the present work is to draw attention to the importance of E . coli as a pathogenic organism . This microorganism is an etiologic agent of sporadic cases of diarrhea, hemorrhagic colitis, dysentery, and hemolytic uremic syndromes and outbreaks . Diarrheic E . coli manifestations occur mainly among infants, and deep knowledge and understanding of this microorganism are crucial to better epidemiologic surveillance. J Pharmacol Toxicol Methods, 2002 Jan-Feb, 47(1), 53 - 8 A simple method to study the activity of natural compounds on the chemiluminescence of neutrophils upon stimulation by immune complexes; Lucisano-Valim YM et al.; INTRODUCTION: Neutrophils (PMNs) are the main effector cells involved in the immune response to microorganisms . However, in various noninfectious states, such as autoimmune and immune complex (ICs) diseases, ICs are found to be deposited in various organs, leading to recruitment and activation of PMNs at these sites of deposition . Consequently, reactive oxygen species (ROS) and lysosomal enzymes are extensively released by activated PMNs into the extracellular milieu, leading to host tissue injury . METHODS: In the present study, we discuss some experimental conditions of a luminol-enhanced chemiluminescence (LECL) assay to study the effect of natural compounds on the production of ROS by rabbit PMNs stimulated with precipitated ICs . Moreover, we evaluated the activities of quercetin and 7-allyloxycoumarin on this ROS-producing system and their toxicity to PMNs . RESULTS: Both compounds had concentration-dependent inhibitory effects on LECL . Quercetin at concentration of 5 micromol/l inhibited 94.5+/-1.0% of LECL, whereas 7-allyloxycoumarin at concentration of 200 micromol/l inhibited 53.8+/-2.4% of LECL . Neither compound was toxic to PMNs under the tested conditions . DISCUSSION: The proposed method may be useful for the screening of nontoxic compounds that can modulate ROS production by IC-stimulated PMNs . Special attention should be devoted to natural compounds from higher plants, since their potential as sources of new drugs is still largely unexplored. Crit Rev Microbiol, 2002, 28(3), 187 - 248 Macrophage-mediated innate host defense against protozoan parasites; Stafford JL et al.; Macrophages are immune cells that play a pivotal role in the detection and elimination of pathogenic microorganisms . Macrophages possess a variety of surface receptors devoted to the recognition of non-self by discriminating between host and pathogen-derived structures . Recognition of foreign microorganisms by the macrophage ultimately results in phagocytosis and the eventual destruction of microorganisms by lysosomal enzymes, toxic reactive oxygen and nitrogen intermediates, and/or nutrient deprivational mechanisms . However, protozoan parasites such as Toxoplasma gondii, Trypanosoma cruzi, and Leishmania spp., parasitize macrophages, utilizing them as a host cell for their growth, replication, and/or maintenance of their life cycles . The protozoan parasites of the genus Leishmania are unique in that their intracellular replication in the host is predominantly restricted to a single cell type, the macrophage . This review focuses on the cellular processes involved in macrophage-mediated host defense against protozoan parasites, from the initial host-parasite interactions that mediate recognition to the mechanisms employed by macrophages to destroy and eliminate the pathogen . As an example model system of experimental study, we describe in more more detail the cellular interactions between macrophages and the obligate intracellular parasite of mammalian macrophages, Leishmania spp. Ying Yong Sheng Tai Xue Bao, 2002 Jul, 13(7), 901 - 5 {Bio-active substances derived from marine microorganisms}; Liu Q et al.; Marine microorganisms, which are taxonomically diverse and genetically special, have powerful potential in producing novel bio-active substances . This article summarized research progress in this respect . The results showed that marine bacteria which are main marine microorganism flora can produce rich kinds of bio-active substances and that even though marine actinomycetes and marine fungi are not as many as marine bacteria in species and quantity, they should be paid no less attention about their bio-active substances . Besides, present research are limited to those marine microorganisms which are easily cultured . One of the future research trends will be focused on bio-active substances derived from non-culturable marine microorganisms. Eur J Intern Med . 2002 Oct;13(7):412. C-reactive protein: history and revival; Ablij H et al.; C-reactive protein (CRP) is the prototype acute-phase protein, which can increase up to 1000-fold after the onset of a stimulus . Aside from its disputed role as a marker of infection and/or inflammation in daily clinical practice, the protein has a wide variety of biological properties and functions . Due to its opsonizing abilities and its capability to activate human complement, CRP plays an important role in the innate host defense against different microorganisms, such as bacteria and fungi . The same opsonophagocyting properties can lead to clearance of host cell material, including nuclear constituents . Inflammation is one of the cornerstones in the etiology and pathogenesis of atherosclerosis, which led to worldwide attention being focused on CRP and its role in the process of atherosclerosis . This role may have a dual character . First, CRP levels reflect the 'burden' of inflammation within atherosclerotic lesions, thus reflecting the grade of vulnerability and instability of the plaques . For this reason, an increased level of the protein may be a prelude to rupture of the plaque and, thus, to occlusive arterial disease . Secondly, CRP may play an active role in the atherosclerotic process . CRP plays a role in the expression of different adhesion molecules on endothelial cells and the protein is able to activate human complement within the plaque . Furthermore, the recent discovery of local production of CRP and complement proteins within the plaque suggests an active role for the protein in the inflammatory cascade . Whatever the role for CRP in the atherosclerotic process, it has been proven that an elevated CRP level, with a cut-off point of approximately 3 mg/l, is associated with an increased risk of occlusive arterial disease, especially acute coronary syndromes. Int J Food Microbiol, 2002 Nov 15, 79(1-2), 27 - 34 Bacterial membranes: the effects of chill storage and food processing . An overview; Russell NJ; The shelf life of food is extended by refrigeration because the metabolic processes of food-associated microorganisms are slowed by the lowered temperature . Nonetheless, cold-adapted psychrotrophic food-poisoning and food-spoilage bacteria remain a concern because they possess cold-adapted proteins and membrane lipids that facilitate growth at low temperatures . The use of membrane-disrupting novel preservation techniques, such as ultrasound, high hydrostatic pressure or pulsed electric field, offer the potential for an extension of shelf life . This review considers the interacting and potentially synergistic effects of chill storage or mild heat treatment on membrane properties, with the disruptive effects of membrane-targeted physical treatments. Appl Microbiol Biotechnol, 2002 Oct, 60(1-2), 1 - 11 Epub 2002 Aug 24. Metabolic engineering towards biotechnological production of carotenoids in microorganisms; Lee PC et al.; Carotenoids are important natural pigments produced by many microorganisms and plants . Traditionally, carotenoids have been used in the feed, food and nutraceutical industries . The recent discoveries of health-related beneficial properties attributed to carotenoids have spurred great interest in the production of structurally diverse carotenoids for pharmaceutical applications . The availability of a considerable number of microbial and plant carotenoid genes that can be functionally expressed in heterologous hosts has opened ways for the production of diverse carotenoid compounds in heterologous systems . In this review, we will describe the recent progress made in metabolic engineering of non-carotenogenic microorganisms for improved carotenoid productivity . In addition, we will discuss the application of combinatorial and evolutionary strategies to carotenoid pathway engineering to broaden the diversity of carotenoid structures synthesized in recombinant hosts. Exp Nephrol, 2002, 10(5-6), 408 - 20 Mixed bone marrow or mixed stem cell transplantation for prevention or treatment of lupus-like diseases in mice; Good RA et al.; Scientific analyses fortified by interpretations of immunodeficiency diseases as 'experiments of nature' have revealed the specific immune systems to be comprised of T cells subserving cell-mediated immunities plus B cells and plasma cells which produce and secrete antibodies . These two separate cellular systems regularly interact with each other to produce a coordinated defense which permits mammals to live within a sea of microorganisms that threaten the integrity and the survival of individuals . We have shown that bone marrow transplantation (BMT) can be used as a form of cellular engineering to construct or reconstruct the immune systems and cure otherwise fatal severe combined immunodeficiency . When severe aplastic anemia complicated the first BMT which was performed to cure a fatal severe combined immunodeficiency, a second BMT cured for the first time a complicating severe aplastic anemia . Subsequently, BMT has been used effectively to treat some 75 otherwise fatal diseases such as resistant leukemias, lymphomas, inborn errors of metabolism, and genetic anomalies of the hematopoietic development such as sickle cell anemia, thalassemia, congenital neutropenias, and many other diseases . More recently, we have employed BMT in mice both to cure and cause autoimmunities, and, together, these experiments showed that autoimmunities actually reside in the hematopoietic stem cells . We have also found that mixed BMT or mixed hematopoietic stem cell transplantation (HSCT) can be used to prevent and cure the most complex autoimmunities such as those occurring in BXSB mice and in (NZW x BXSB)F1 W/BF1 mice . Untreated, the former develop fulminating lethal glomerulonephritis plus numerous humoral autoimmunities . Mice of the (W/B)F1 strain develop autoimmune thrombocytopenic purpura, coronary vascular disease with myocardial infarction, glomerulonephritis, and numerous autoantibodies . All of these abnormalities are prevented or cured by mixed syngeneic (autoimmune) plus allogeneic (normal healthy) BMT or mixed peripheral blood HSCT . Thus, the most complex autoimmune diseases can be prevented or cured in experimental animals by mixed syngeneic plus allogeneic BMT or HSCT which produce stable mixed chimerism as a form of cellular engineering . Ann N Y Acad Sci, 2002 Oct, 969, 48 - 50 Public health considerations in human consumption of wild game; Ahl AS et al.; The role of a few microorganisms, like Brucella and Mycobacterium and certain parasites of food animals, in causing human disease has been recognized for a hundred years . By the 1990s, other microorganisms derived from food animals were recognized as contributing to human illness . Handling and/or consumption of wild game may result in human exposure to novel microorganisms; these unrecognized or unknown agents or diseases in wild species may cross into humans and cause "new" diseases with which we are not familiar. J Agric Food Chem, 2002 Oct 23, 50(22), 6628 - 34 Plasmin system and microbial proteases in milk: characteristics, roles, and relationship; Nielsen SS; Proteolysis of milk proteins can be attributed to both native proteases and the proteases produced by psychrotrophic bacteria during storage of fresh raw milk . These proteases cause beneficial or detrimental changes, depending on the specific milk product . Plasmin, the major native protease in milk, is important for cheese ripening . Milk storage and cheese-making conditions can affect the level of plasmin in the casein and whey fractions of milk . A microbial protease from a psychrotrophic microorganism can indirectly increase plasmin levels in the casein curd . This relationship between the plasmin system and microbial proteases in milk provides a means to control levels of plasmin to benefit the quality of dairy products . This paper is a short review of both the plasmin system and microbial proteases, focusing on their characteristics and relationship and how the quality of dairy products is affected by their proteolysis of milk proteins. Can J Microbiol, 2002 Aug, 48(8), 717 - 27 Predicting survival of a genetically engineered microorganism, Pseudomonas chlororaphis 3732RN-L11, in soil and wheat rhizosphere across Canada with linear multiple regression models; Edge TA et al.; Pseudomonas chlororaphis 3732RN-L11 survival rates in soil and wheat rhizosphere were measured using intact soil core microcosms representing 23 sites across Canada . Linear multiple regression (LMR) models were developed to predict the survival rate of this genetically engineered microorganism (GEM) as a function of soil parameters measured at the time of microcosm inoculation . LMR models were tested by comparing their predicted survival rates with observed survival rates from environmental introductions of the GEM by Gagliardi et al . (2001) at five field sites across Canada over two years . No soil parameter (e.g., % clay) was highly correlated with GEM survival rates in soil or wheat rhizosphere . Total fungal colony-forming units (CFUs), % soil titanium (positive correlations), and % soil magnesium (negative correlation) were found to be the best LMR predictors of GEM survival rates in soil over two years . Total soil bacterial CFUs, nitrate, % soil potassium (positive correlations), and exchangeable magnesium (negative correlation) were found to be the best LMR predictors of GEM survival rate in wheat rhizosphere over two years . While LMR models were statistically significant, they were unable to reliably predict the survival rate of the GEM in field trial introductions . The results indicate that there can be considerable uncertainty associated with predicting GEM survival for multi-site environmental introductions. Vestn Ross Akad Med Nauk, 2002, (9), 38 - 40 {Environmental and occupational problems in the utilization of industrial and home waste}; Rusakov NV et al.; One of the acute hygienic problems of today is handling industrial and consumer wastes stored in Russia in the amount of 30 milliard tons; 30 million of solid garbage and 120 million tons of industrial wastes being formed . There are 4 garbage-handling and 10 garbage disposal plants built in the country . A third of them don't operate now . An absence of the initial selection in garbage collection causes the situation when only 3% of solid garbage is being processed . There is no state unified wastes recycling and utilizing system, no training of personnel to work out special technologies and do this work . Special attention should be given to medical wastes dangerous for the epidemiological situation, since they contain pathogenic microorganisms and helminth eggs; they may as well be contaminated by toxic and radioactive chemicals . A complex solution of the problems of industrial and consumer wastes handling is of vital importance for human health protection and protection of environment. Tsitol Genet, 2002 Jul-Aug, 36(4), 30 - 6 {Study of microorganism genome DNA by atomic force microscopy}; Lymans'kyi OP et al.; The potential of atomic force microscopy (AFM) for the investigation of peculiarities of microorganisms genome structure is demonstrated . AFM images of phage lambda DNA linear molecules and supercoiled mica in buffer solution was imaged in air . New experimental method of DNA stretching based on using amino-modified mica with a decreased surface density of active amino-groups is proposed . Stretched molecules of phage lambda DNA were imaged by AFM. Drug Chem Toxicol, 2002 Nov, 25(4), 459 - 72 Effect of lead and arsenic on murine macrophage response; Sengupta M et al.; Splenic macrophages are highly efficient in trapping and concentrating foreign substances carried in the blood and also the major sites where antibodies are synthesised and from where they are released into the circulation . Lead and Arsenic as environmental agents are considered to be high priority toxic substances largely due to their carcinogenic potentials in humans . However, these heavy metals as carcinogens remain an enigma because while they are definitely active in humans, carcinogenesis in the rodent model has never been convincingly demonstrated . Although macrophages are predominantly recruited to the site of inflammation during inflammatory distress as well as in immune response, nothing is known about the interaction of lead and arsenic with macrophages and their possible role in immunotoxicologic effect . In the present study it is reported that in vivo lead acetate treatment (10 mg/kg body wt) inhibits the cell adhesion property and alters the cell morphology in the splenic macrophages . Results show that there is a significant decrease in alkaline phosphatase release in lead treated macrophages (6.7 +/- 0.88 IU/100 mL) with respect to control (14.17 +/- 0.18) . In vivo exposure of sodium arsenite (0.5 mg/kg body wt) also decreases phagocytic activity for ingestion and digestion of exogenous antigens, such as whole microorganism, as evident from the phagocytic index, 11555.55 +/- 62.86 (in control) to 5555.5 +/- 1571.33 in arsenic treated cells . Arsenic exposed cells release 8.15 +/- 0.05 microM nitric oxide, whereas control cells release 10.95 +/- 0.15 microM of nitric oxide, which is also identical following LPS stimulation . Results show that the functional integrity of the target cell is also decreased after arsenic exposure as obtained from the percentage of DNA fragmentation . A greater percentage of DNA fragmentation upon arsenic treatment (43.1 +/- 0.05%) with respect to control (14.9 +/- 0.34%) indicates that arsenic induces apoptosis . In immune cells which are rapidly proliferating and differentiating, inhibition of these heavy metal induced functions may result in similar degree of toxicity and lead to diseased state. Klin Khir, 2002 Jul, (7), 21 - 3 {Colonic sanation in peritonitis caused by acute destructive cholecystitis}; Karliichuk OO; Specific and quantitative microorganisms composition of colon cavity in 53 patients with an acute biliary peritonitis was investigated . Pathogenetic substantiated method of colonic sanation for treatment of peritonitis, caused by an acute destructive cholecystitis was elaborated . There was established, that in occurrence of the biliary genesis peritonitis the disorders of microbic intestinal cenosis and its colonizational resistance play an important role . Application of proposed method of colonic sanation allowed to eliminate its disorders and to improve result of treatment of patients. Prep Biochem Biotechnol, 2002 Aug, 32(3), 217 - 37 Biocatalyst-adsorbant systems: a viable alternative to proteolytic processes in solution; Furtado S et al.; Proteolytic biocatalysts were adsorbed and stabilized using alumina as a support medium . Two biocatalyst-adsorbant systems were prepared with different physical characteristics of the adsorbant: alumina powder and alumina pearls . Direct adsorption onto the support medium has the main advantage, over other fixation methods, that preliminary steps are not required for a good interaction between the support and the biocatalyst . Proteases were adsorbed and stabilized without modifying or sterically hindering their active sites . Parameters affecting adsorption (pH, temperature, ionic strength) were varied so as to optimize adsorption conditions . Operational viability of the immobilized biocatalysts was demonstrated, taking into account the rate of desorption, resistance to microbial attack, and stability during storage . Desorption in water was studied in batch and continuous-flow processes, at various flow rates . The systems also proved to be resistant to microorganisms . Tests for stability during storage found the systems' activity remained constant after 60 days, and they performed better than biocatalysts in solution . Proteolysis of a solution of g per litre of azocasein was carried out in continuous-flow and batch modes, using our biocatalyst-adsorbant systems we prepared . In all cases, free amino group concentrations were around 2.5 times greater after treatment with biocatalyst-adsorbants than they were in the starting solution. J Investig Allergol Clin Immunol, 2002, 12(2), 73 - 9 Chlamydia pneumoniae infection and its role in asthma and chronic obstructive pulmonary disease; Clementsen P et al.; Chlamydia pneumoniae (CP) is a common cause of respiratory tract infections, and several studies have asked whether it may play a pathogenic role in connection with bronchial asthma and chronic obstructive pulmonary disease (COPD) . Evidence that CP infection is associated with these diseases is a cardinal item . However, evaluation of CP infection is hampered by difficulties in obtaining agreement on the definition of a gold standard . In the literature, serology is based on different cutoff points of antibody titres, which complicates the definition of CP seropositive findings and the classification of acute infection, chronic and past infection . In connection with acute and chronic infection, it is important to demonstrate the presence of CP by culture or polymerase chain reaction (PCR) in the respiratory tract, especially in the lower airways . Often, the results of serology is not associated with the findings by culture or PCR testing, which may involve the risk of inconclusive evidence . Evaluation of a possible presence of CP by clinical improvement after treatment with antibiotics is difficult since uncontrolled studies have been used and other microorganisms are also affected by antibiotics . Furthermore, many patients improve without antibiotics, and improvement has also been observed in patients remaining culture positive after treatment with antibiotics . It should also be noted that the antiinflammatory effects of antibiotics may improve the clinical status of patients . Despite these obstacles, studies point to the possibility that in some patients acute CP infections may lead to acute exacerbations of bronchial asthma . Whether a persistent CP infection contributes to chronic asthma or severe COPD, or whether it incites the diseases in previously healthy individuals is a question for further studies . Whether a causal relationship exists between CP infection and obstructive pulmonary disease or whether these patients are more susceptible to CP infection is unknown . Nevertheless, a cooperative role of CP in the proinflammatory mechanisms involved in these diseases remains to be examined since cellular studies show that CP stimulates the production and expression of cytokines, chemokines and adhesion molecules, actions that may amplify and prolong the inflammation. Environ Toxicol Chem, 2002 Oct, 21(10), 2099 - 107 Prediction of uptake dynamics of persistent organic pollutants by bacteria and phytoplankton; Del Vento S et al.; Phytoplankton and bacteria play an important role on the biogeochemical cycles of persistent organic pollutants (POPs) . However, experimental data and quantitative knowledge of the kinetics of uptake and depuration of most POPs by bacteria and phytoplankton are scarce . In the present paper, a procedure to predict the sorption kinetics to bacteria and phytoplankton is developed . The prediction method is the combination of a mechanistic model for sorption and quantitative structure-activity relationships relating bioconcentration factors and membrane permeability to the chemical physical-chemical properties . The model consists of two compartments where the first compartment is the cellular surface and the second compartment is the cell biomass or matrix . Equations for estimating uptake and depuration rate constants into the matrix and adsorption and desorption rate constants onto the surface are obtained . These expressions depend on the physical-chemical properties of the chemical, the environmental temperature, the microorganism size, and species-specific quality of organic matter . While microorganism shape has a secondary influence on uptake dynamics, microorganism size and chemical hydrophobicity arise as the key factors controlling the kinetics of POP incorporation into bacteria and plankton . Uptake, depuration, adsorption, and desorption rate constants are reported for POPs such as polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), polychlorinated dioxins and furans (PCDD/Fs), and POPs of emerging concern, such as polybrominated diphenyl ethers (PBDEs) . Finally, implications of uptake and depuration dynamics on the biogeochemical cycling of POPs are discussed. Clin Chest Med, 2002 Sep, 23(3), 569 - 83 Pathogenesis of nontuberculous mycobacteria infections; McGarvey J et al.; M avium is a microorganism well adapted to living in the environment and in different hosts . During the past 15 years, a substantial amount of information has been accumulated about the mechanisms used by M avium to cross the host's mucosal barrier, replicate inside cells, circumvent the host's immune response, and persist inside the host . It turns out that M avium is a fascinating pathogen after all . The increasing knowledge about M avium pathogenesis may one day provide means for a more effective prophylaxis as well as for treatment of the infection. EMBO Rep, 2002 Oct, 3(10), 938 - 43 ABC transporters: one, two or four extracytoplasmic substrate-binding sites? van der Heide T, Poolman B. Two families of ATP-binding cassette (ABC) transporters in which one or two extracytoplasmic substrate-binding domains are fused to either the N- or C-terminus of the translocator protein have been detected . This suggests that two, or even four, substrate-binding sites may function in the ABC transporter complex . This domain organization in ABC transporters, widely represented among microorganisms, raises new possibilities for how the substrate-binding protein(s) (SBPs) might interact with the translocator . One appealing hypothesis is that multiple substrate-binding sites in proximity to the entry site of the translocation pore enhance the transport capacity . We also discuss the implications of multiple substrate-binding sites in close proximity to the translocator in terms of broadened substrate specificity and possible cooperative interactions between SBPs and the translocator. Trends Biochem Sci, 2002 Oct, 27(10), 497 - 500 BLUF: a novel FAD-binding domain involved in sensory transduction in microorganisms; Gomelsky M et al.; A novel FAD-binding domain, BLUF, exemplified by the N-terminus of the AppA protein from Rhodobacter sphaeroides, is present in various proteins, primarily from Bacteria . The BLUF domain is involved in sensing blue-light (and possibly redox) using FAD and is similar to the flavin-binding PAS domains and cryptochromes . The predicted secondary structure reveals that the BLUF domain is a novel FAD-binding fold. J Periodontal Res, 2002 Oct, 37(5), 360 - 5 Effects of hippocampal lesioning on experimental periodontitis in Wistar rats; Breivik T et al.; The hippocampus, which is a brain structure involved in learning and memory processes, plays a key role in the feedback regulation of the hypothalamic-pituitary-adrenal (HPA) axis and autonomic sympathetic nervous system, and the subsequent secretion of immuno-modulatory hormones in response to pathogenic microorganisms . Dysregulation of these brain-neuroendocrine-immune regulatory networks, which act in concert to maintain homeostasis, is found to be of critical importance to the host defence against pathogens, as well as susceptibility to diseases, including periodontal disease . The present study was designed to determine the effects of hippocampal lesioning on the progression of periodontitis . Experimental ligature-induced periodontitis was induced in 16 Wistar rats, which were bilaterally lesioned in their hippocampal region with an aspiration technique that is well documented to impair learning and memory, as well as in 15 sham-operated control rats . The disease progression was evaluated radiographically and histometrically . The results revealed that the hippocampal lesioned rats developed significantly more destruction of the periodontium than did the sham-operated controls . This finding supports recent studies that indicate that inappropriate brain-neuroendocrine regulation of inflammatory responses to infectious agents may play an important role in disease susceptibility and progression. Science, 2002 Oct 4, 298(5591), 216 - 8 Excess polymorphisms in genes for membrane proteins in Plasmodium falciparum; Volkman SK et al.; The detection of single-nucleotide polymorphisms in pathogenic microorganisms has normally been carried out by trial and error . Here we show that DNA hybridization with high-density oligonucleotide arrays provides rapid and convenient detection of single-nucleotide polymorphisms in Plasmodium falciparum, despite its exceptionally high adenine-thymine (AT) content (82%) . A disproportionate number of polymorphisms are found in genes encoding proteins associated with the cell membrane . These genes are targets for only 22% of the oligonucleotide probes but account for 69% of the polymorphisms . Genetic variation is also enriched in subtelomeric regions, which account for 22% of the chromosome but 76% of the polymorphisms. Clin Microbiol Rev, 2002 Oct, 15(4), 545 - 63 Atopic dermatitis and fungi; Faergemann J; Atopic dermatitis (AD) is a chronic, itching, inflammatory skin disease which is associated with asthma and/or hay fever and a familial occurrence of these conditions . Genetic factors are important in the development of AD, but the exact hereditary pathway is still unknown . Dry skin and the weakened barrier function in patients with AD is very important for the patient's reactions to irritants and other external trigger factors including microorganisms . The standard treatments are topical corticosteroids, topical immunomodulating agents, and emollients . If AD cannot be controlled by this type of treatment, systemic immunomodulating agents may be used . UVB, UVA, or psoralen-UVA may also be used for widespread severe lesions . However, some patients do not respond to these standard treatment, and then it is important to consider the role of microorganisms, house dust mites or food . The role of the Malassezia yeasts in AD, especially AD located to the head and neck region, is now documented in several papers . There are also several papers indicating the role of Candida as an aggravating factor in AD . Patients with AD also develop chronic dermatophyte infections more easily, and patients with AD and chronic dermatophyte infections may show improvement in their AD when treated with antifungal drugs. Adv Drug Deliv Rev, 2002 Oct 4, 54(6), 851 - 61 A strategic approach to vaccine development: animal models, monitoring vaccine efficacy, formulation and delivery; Griffin JF; One of the most pressing issues in animal and human medicine currently is the need to develop new and more effective vaccines, against an ever increasing range of infectious diseases . Traditional vaccines have involved the use of killed microorganisms, live attenuated cultures or antigenic extracts . In spite of extensive research, there have been very few newly developed vaccines for humans or animals, over the past 20 years . Recent developments in our understanding of the pathways of immunity required to produce protection against different infections, allow immunological principles to be incorporated into the design new and better vaccines . This paper outlines the key variables that need to be taken into consideration when developing a new vaccine . Three critical areas will be considered in the review: (1) use of an appropriate experimental animal model of infection; (2) targeting appropriate immune markers for protection; (3) design of optimal vaccine formulation and delivery systems. Biotechnol Prog, 2002 Sep-Oct, 18(5), 994 - 1002 Multiple substrate growth kinetics of Leptothrix discophora SP-6; Yurt N et al.; The growth parameters of Leptothrix discophora SP-6 were quantified on the basis of the steady-state concentrations and utilization rates of pyruvate, dissolved oxygen, and concentration of microorganisms in a chemostat operated at 25 degrees C, pH 7.2, and an agitation rate of 350 rpm . The results showed that the microbial growth was limited by both pyruvate and dissolved oxygen . A combined growth kinetics model using Monod growth kinetics for pyruvate and Tessier growth kinetics for oxygen showed the best correlation with the experimental data when analyzed using an interactive multiple substrate model . The growth kinetics parameters and the respective confidence limits, estimated using the Monte Carlo simulation, were mu(max) = 0.576 +/- 0.021 h(-1), K(sMp) = 38.81 +/- 4.24 mg L(-1), K(sTo) = 0.39 +/- 0.04 mg L(-1), Y(X/p) = 0.150 (mg microorganism mg(-1) pyruvate), Y(X/o) = 1.24 (mg microorganism mg(-1) oxygen), the maintenance factors for pyruvate and oxygen were m(p) = 0.129 (mg pyruvate consumed mg(-1) microorganism h(-1)) and m(o) = 0.076 (mg oxygen consumed mg(-1) microorganism h(-1)), respectively. J Environ Radioact, 2002, 63(2), 187 - 97 Biologically induced Po emission from fresh water; Momoshima N et al.; Behavior of Po in fresh waters was examined in laboratory culture experiments using fresh water collected from a small pool, Xi river and Xiqing lake, showing formation of volatile Po compounds followed by emission to air . Addition of tryptone to the fresh water cultures increased the emission of Po considerably along with a growth of microorganisms, suggesting a connection of chemoheterotrophs to Po emission . Participation of photoautotrophs was also considered because Po emission was increased when NaHCO3 was added to the fresh water cultures . The emission behavior of Po and S in these experiments appeared in different ways . The quantity of Po emitted was comparable to the previous culture experiments (Momoshima, Song, Osaki & Maeda, Environ . Sci . Technol., 35, 2956-2960, 2001) in which artificial culture medium containing 3% NaCl was used and inoculated with sea sediment extract . The biological support for Po emission, thus, would be a general phenomenon in fresh water as well as a seawater environment and is possibly a source for atmospheric Po. Oral Dis, 2002 Sep, 8(5), 254 - 60 Nitric oxide synthesis and severity of human periodontal disease; Batista AC et al.; The expression of the inducible nitric oxide synthase enzyme (iNOS) is a response to an inflammatory stimulus and produces a large amount of nitric oxide (NO), which may act as a cytotoxic molecule against the invading microorganism and may be related to both harmful and beneficial effects to tissues . OBJECTIVE AND MATERIAL AND METHODS: In order to further characterize the presence of NO in human periodontal disease, we undertook a quantitative study of iNOS positive cells in samples of clinically healthy gingival tissues, plaque-induced gingivitis and localized chronic periodontitis using immunohistochemistry . RESULTS: A significant increase in the number of iNOS+ cells mm-2 was found in the samples of the gingivitis and periodontitis compared with those of the control . In all groups most of the polymorphonuclear cells showed intense immunoreactivity for iNOS independent of the disease stage, and the percentage of iNOS+ polymorphonuclear cells increased significantly in periodontal disease when compared with the control . CONCLUSION: Our results indicate that iNOS increases in the presence of periodontal disease . In addition, our findings suggest that polymorphonuclear cells present an additional activation pathway in periodontal disease, expressing significant iNOS and probably representing an important source of NO in human periodontal disease that has not been previously reported. Pol Merkuriusz Lek, 2002 Jun, 12(72), 522 - 5 {Immunological phenomenons connected with etiology and pathogenesis of sarcoidosis}; Mamelka B et al.; Despite of numerous studies on etiology and pathogenesis of sarcoidosis, it has been impossible to explain the reason of disease initiation . The possible beginning of sarcoid process is broken balance of cytokines secreted by T-lymphocytes (Th1 and Th2), which generates various complicated immunological phenomenons . This broken balance of cytokines may be caused by particular antigen in the shape of microorganism or other factor, which disturbs organism's homeostasa. J Dairy Sci, 2002 Sep, 85(9), 2308 - 14 Nucleosides are efficiently absorbed by Na(+)-dependent transport across the intestinal brush border membrane in veal calves; Theisinger A et al.; In previous work, a comparatively high capacity for Na(+)-dependent transport of nucleosides across the intestinal brush border membrane (BBM) was observed in dairy cows, which might be related to digestion of the large amount of nucleic acids present in ruminal microorganisms in the ruminant small intestine . If this were the case, the capacity for Na(+)-dependent intestinal nucleoside transport should be much lower in veal calves, in which only small amounts of nucleic acids, nucleotides, and nucleosides reach the small intestine via the milk replacer . To test this hypothesis, we investigated Na(+)-dependent transport of 3H-labeled thymidine and guanosine across the BBM using BBM vesicles (BBMV) isolated from the small intestine of veal calves . In the presence of a transmembrane Na+ gradient both substrates were transported against a concentration gradient . Inhibitory studies showed that thymidine and guanosine are transported by two different transporters with overlapping substrate specificity, one accepting predominantly pyrimidine nucleosides (N2) and one accepting particularly purine nucleosides (N1) . Nucleoside transport was inhibited by glucose along the whole small intestine . Maximal transport rates similar to those in dairy cows were obtained for the proximal, mid-, and distal small intestine . These findings suggest that the high absorptive capacity for nucleosides is a genetically fixed property in the bovine small intestine, which is already present in the preruminant state of veal calves . It may contribute to the high digestibility of nucleic acids observed by others in veal calves receiving milk replacer supplemented with RNA . Its main function may be the efficient absorption of nucleosides resulting from the digestion of nucleic acids associated with desquamated enterocytes . Due to the limited de novo synthesis of nucleotides in enterocytes intracellular uptake of nucleosides across the BBM may contribute to nucleic acid synthesis in enterocytes and thus may have a trophic effect on the intestinal epithelium. J Basic Microbiol, 2002, 42(5), 327 - 36 Molecular analysis of arylalcohol dehydrogenase of Coriolus versicolor expressed against exogenous addition of dibenzothiophene derivatives; Ichinose H et al.; Using the differential display reverse-transcriptional polymerase chain reaction (DDRT-PCR) technique, several cDNA fragments were isolated as chemical stress responsive genes from the white-rot basidiomycete, Coriolus versicolor, exposed to either 4-methyldibenzothiophene-5-oxide (4MDBTO) or dibenzothiophene-5-oxide (DBTO) . A database search on deduced amino acid sequences of cDNAs revealed that they showed a high similarity with various proteins from other organisms . These results strongly suggested that cell responding systems might be involved in the fungal metabolism of exogenous chemicals by C . versicolor . One of the significantly up-regulated cDNA fragments by MDBTO, DD16gc, showed a high similarity to arylalcohol dehydrogenases (AADs) from several microorganisms . The full-length cDNA sequence of the DD16gc determined by 5' rapid amplification of cDNA ends method revealed that the gene consisted 1,295 nucleotide and poly(A) tail, encoding 394 amino acids in an open reading frame . The deduced protein showed a remarkable homology to AAD from Phanerochaete chrysosporium (66% identity) and to that from Saccharomyces cerevisiae (54% identity) . The AAD gene was specifically transcripted under chemically-stressed conditions by 4MDBTO, suggesting that the enzyme encoded by the stress responsive gene may play an important role in the fungal conversion of 4MDBTO or its metabolic product(s). Br J Community Nurs . 2002 Sep;:8, 10, 12, passim. To use or not to use? The debate on the use of antiseptics in wound care; Scanton E et al.; The use of antiseptics for wound care has long been controversial . Antiseptics have a broad spectrum of activity that, in addition to making them effective against a broad range of microorganisms, makes them toxic to living human tissue . Consequently antiseptics gradually fell out of favour following the introduction of systemic antibiotics . In recent years, however, there has been renewed interest in antiseptics as a result of the evolution of antibiotic-resistant bacteria, and considerable funds are being invested in the development of modern antiseptic dressings . This article aims to discuss some of these issues and set them in a practical context so that readers recognize the complexities of decision-making where antiseptics are concerned . This in turn should enable them to make a better-informed choice for their patients or, where appropriate, seek expert advice. Eur J Gastroenterol Hepatol, 2002 Oct, 14(10), 1113 - 8 Bleeding peptic ulcers and presence of Helicobacter pylori by various tests: a case-control study; Castillo-Rojas G et al.; BACKGROUND: Virulence factors of Helicobacter pylori are associated with peptic ulcer disease and may be also associated with bleeding peptic ulcers (BPU) . AIM: To determine whether H . pylori and/or the cytotoxin-associated gene (cagA) can increase the risk of bleeding in peptic ulcers . PATIENTS: Sixty-seven patients were studied . Thirty had BPU, 20 had non-bleeding peptic ulcers (NBPU), and 17 were control subjects (NPU) . METHODS: The prevalence of H . pylori was assessed by the urease fast test, histological examination, serology, and 16S ribosomal RNA and cagA gene amplification by polymerase chain reaction (PCR) . RESULTS: Histology and PCR showed greater sensitivity for diagnosis of H . pylori under bleeding circumstances when compared with other tests . Association of H . pylori was greater in the NBPU group (odds ratio {OR} 4.91, P = 0.06) than in the BPU group (OR 1.27, P = NS) when compared with the control group . When the BPU and NBPU groups were compared, H . pylori was found more often in the NBPU group (OR 0.26, P < 0.10 ) . The cagA-positive gene showed a similar distribution in the three groups . The titres for anti-CagA immunoglobulin A (IgA) antibodies were higher in NBPU patients (83%) than in BPU or control patients . Furthermore, anti-urease immunoglobulin G (IgG) was detected more frequently among BPU and NBPU patients . CONCLUSIONS: NBPU patients had the highest prevalence of H . pylori by PCR . It seems unlikely that either H . pylori or the cagA-positive gene act as significant risk factors for bleeding in peptic ulcers . The lower prevalence of the microorganism among patients who bleed cannot be explained as an artificial finding. Rev Immunogenet, 2000, 2(4), 477 - 91 Identifying cytotoxic T cell epitopes from genomic and proteomic information: "The human MHC project."; Lauemoller SL et al.; Complete genomes of many species including pathogenic microorganisms are rapidly becoming available and with them the encoded proteins, or proteomes . Proteomes are extremely diverse and constitute unique imprints of the originating organisms allowing positive identification and accurate discrimination, even at the peptide level . It is not surprising that peptides are key targets of the immune system . It follows that proteomes can be translated into immunogens once it is known how the immune system generates and handles peptides . Recent advances have identified many of the basic principles involved . The single most selective event is that of peptide binding to MHC, making it particularly important to establish accurate descriptions and predictions of peptide binding for the most common MHC variants . These predictions should be integrated with those of other steps involved in antigen processing, as these become available . The ability to translate the accumulating primary sequence databases in terms of immune recognition should enable scientists and clinicians to analyze any protein of interest for the presence of potentially immunogenic epitopes . The computational tools to scan entire proteomes should also be developed, as this would enable a rational approach to vaccine development and immunotherapy . Thus, candidate vaccine epitopes might be predicted from the various microbial genome projects, tumor vaccine candidates from mRNA expression profiling of tumors ("transcriptomes") and auto-antigens from the human genome. J Biochem Mol Biol, 2002 Sep 30, 35(5), 482 - 7 Endotoxins of enteric pathogens are chemotactic factors for human neutrophils; Islam LN et al.; Early activation of human peripheral blood polymorphonuclear neutrophils is characterized by their morphological changes from spherical to polarized shapes . The endotoxins from enteric pathogens (S . dysenteriae type 1, V . cholerae Inaba 569B, S . typhimurium, and K . pneumoniae) were assessed by their ability to induce morphological polarization of the neutrophils as measures of early activation . Phagocytic activity, adhesion, chemokinetic locomotion, and nitroblue tetrazolium (NBT) dye-reduction ability measured the later activation of the cells . Neutrophils showed distinct morphological polarization in suspension over a wide range of concentrations of these endotoxins when were compared with those that were induced by the standard chemotactic factor, N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) . It was discovered that all of the endotoxins induced locomotor responses in neutrophils in suspension that were dose- and time-dependent . The optimum concentration for the endotoxins of S . dysenteriae, V . cholerae, and K . pneumoniae was 1 mg/ml in which 71, 69, and 66% of the neutrophils were polarized . However, the S . typhimurium dose was 2 mg/ml in which 50% of the cells responded . Neutrophils that were stimulated with endotoxins also showed increased random locomotion (p<0.005) through cellulose nitrate filters, but an enhanced adhesion of the cells to glass surfaces (p<0.03) . These are important functions of these cells to reach and phagocytose damaged cells, as well as invading microorganisms . Interestingly, the endotoxins had a highly-significant inhibitory effect upon the proportions of neutrophils phagocytosing opsonized yeast (p<0.01) with a small number of yeast that were engulfed by the cells (p<0.02) . Further, endotoxin-treated cells showed an enhanced ability to reduce NBT dye (p<0.03) . Therefore, we concluded that endotoxins of enteric pathogens are neutrophil chemotactic factors. J Biochem (Tokyo), 2002 Oct, 132(4), 621 - 7 Expression and regulation of the gene for arginase I in mouse salivary glands: requirement of CCAAT/enhancer-binding protein alpha for the expression in the parotid gland; Akiba T et al.; Arginase in salivary glands is potentially involved in the synthesis of proline, glutamate, and polyamines that play specific physiological roles in the glands, and also in depletion of arginine in the oral cavity to protect teeth from microorganisms . We detected protein and mRNA for the type I isoform of arginase in mouse salivary glands . Enzymes of the arginine-biosynthetic pathway were also detected . Immunohistochemical analysis revealed that arginase I was enriched in the striated duct, and was also present in the acinus, demilune and granulated duct . Mice with targeted disruption of the gene for C/EBPalpha, which is a transcription factor essential for expression of the arginase I gene in the liver, showed dramatically reduced immunoreactivity for arginase I in the parotid gland but not in the submandibular and sublingual glands . Therefore, C/EBPalpha is specifically required for expression of the arginase I gene in the parotid gland. Urologiia, 2002 Jul-Aug, (4), 39 - 43 {Status and development of the kidney after surgical treatment of hydronephrosis in children}; Cheskis AL et al.; Development of the kidneys after correction of hydronephrosis was studied in 40 patients operated at the age of 8 months-12 years . Follow-up lasted 10-25 years . X-ray control showed efficiency of correction of the uretheropelvic segment (UPS) . Roentgenoplanimetric study demonstrated a uniform 50-100% increase in renal area and its parenchyma in 28 patients (32 kidneys) . In 9 patients parenchyma increased 2.5-4-fold, renal area by 50-100% . In three cases parenchyma diminished by 3-10% in growth of the kidney by 24-80% . This evidenced of nephrosclerosis progress . UPS correction favoured renal growth and development . There were only few cases when correction of UPS was positive but nephrosclerosis aggravated . Development of the kidney may depend on many factors such as preoperative condition of the kidney, severity of UPS obstruction, the patient's age, duration and severity of pyelonephritis, dysplasia and age-related immaturity of renal parenchyma, immune status of the organism, microorganism virulence, etc . But decisive was the condition of renal parenchyma before the operation . In 3 patients with progressive nephrosclerosis renal parenchyma affection was additionally characterized by advanced sclerosis and inflammatory infiltration of the intertitium which ended in a poor outcome . The above observations suggest the necessity of long-term postoperative follow-up of the patients to design effective measures aimed at the treatment and prevention of nephrosclerosis. Curr Opin Crit Care, 2002 Oct, 8(5), 461 - 4 Prevention of infection in the intensive care unit: current advances and opportunities for the future; Lorente C et al.; Recent studies have contributed to our understanding of the risk factors and the impact of nosocomial infections in the ICU, allowing a more rational approach to the prevention of such infections . Ventilator-associated pneumonia, bloodstream infections, and outbreaks all occur in the presence of artificial devices . High antibiotic pressure, prolonged hospitalization, and the presence of comorbidities facilitate the selection of multiresistant strains in the ICU setting . In clinical practice, prevention is the more effective investment to reduce costs . Potential measures of control should focus on the patient, the microorganisms, and the device . A number of recent studies addressing these issues have been published and will be reviewed in this article. Comput Biol Med, 2002 Nov, 32(6), 529 - 49 Analogy between temperature-dependent and concentration-dependent bacterial killing; Neef C et al.; In this article an analogy between temperature-dependent and concentration-dependent bacterial killing is described . The validation process of autoclaves uses parameters such as reduction rate constant k, decimal reduction time D and resistance coefficient z from an imaginary microorganism to describe the sterilization process . Total lethality of the process is calculated as the integral of the lethality (a function of the temperature) over time.In the case of concentration-dependent killing-i.e . using antibiotic drugs-the k-value is not necessarily a constant; it is the difference between growth and killing of the microorganism . Equations are derived for the decimal reduction time D and resistance coefficient z . Pharmacodynamic models of tobramycin, ciprofloxacin and ceftazidime are used to demonstrate that there is an optimal concentration for all three drugs: C(opt-tobra)=3.20 MICmg/l, C(opt-cipro)=3.45 MICmg/l and C(opt-cefta)=1.35 MICmg/l. J Ind Microbiol Biotechnol, 2002 Oct, 29(4), 204 - 8 Deep-sea hydrothermal vents: a new source of innovative bacterial exopolysaccharides of biotechnological interest? Guezennec J. Polysaccharides and, in particular, microbial polysaccharides represent a class of important products of growing interest for many sectors of industry . Although many known marine bacteria produce exopolysaccharides (EPS), continuation in looking for new polysaccharide-producing microorganisms is promising . Hydrothermal deep-sea vents could be a source of novel EPS as indicated by the screening of a number of mesophilic heterotrophic bacteria recovered from different locations . Although originating from such extreme environment, some bacteria were shown to biosynthesize innovative EPS under laboratory conditions . Their specific rheological properties either in the presence or absence of monovalent and divalent ions, biological activities, metal binding capabilities, and novel chemical composition mean that these EPS are expected to find many applications in the near future. Nat Biotechnol, 2002 Oct, 20(10), 1035 - 9 Epub 2002 Sep 23. Engineering hypervirulence in a mycoherbicidal fungus for efficient weed control; Amsellem Z et al.; Agents proposed for biocontrol of major weeds in arable row-crop agriculture have not met expectations because an evolutionary balance has developed between microorganism and weed, even when the mycoherbicide is used inundatively at very high levels (>10(4)spores/cm<(2)) . Sufficient virulence can be achieved by transferring genes to the microorganism, tipping the evolutionary balance . Virulence was increased ninefold and was more rapidly effected; furthermore, the requirement for a long duration at high humidity was decreased by introducing NEP1 encoding a phytotoxic protein, to an Abutilon theophrasti-specific, weakly mycoherbicidal strain of Colletotrichum coccodes . The parent strain was at best infective on juvenile cotyledons of this intransigent weed . The transgenic strain was lethal through the three-leaf stage, a sufficient time window to control this asynchronously germinating weed . Strategies of coupling virulence genes with fail-safe mechanisms to prevent spread (due to broadened host range) and to mitigate transgene introgression into crop pathogens could be very useful in the biocontrol of major weeds in row crops. Neurosci Res, 2002 Oct, 44(2), 207 - 12 Nestin and small heat shock protein expression on reactive astrocytes and endothelial cells in cerebral abscess; Ha Y et al.; Cerebral abscess results in an extensive capsular formation, which is an important limiting barrier for the spread of microorganism . Reactive astrocytes and endothelial cells are major cellular components of the abscess capsule together with fibrocytes . Molecular pathogenesis that results in the migration and proliferation of these cells remain speculative . Intermediate filament (IF) nestin and small heat shock proteins (sHSP) are developmentally regulated protein . In this study, we found that nestin expression is re-induced in not only reactive astrocytes, but also in endothelial cells in the gliotic area of the capsule . These nestin expressing astrocytes and endothelial cells also expressed sHSPs, such as heat shock protein 27 (HSP27) and alphaB-crystalline . These results indicated that embryonic reversions of cytoskeletal proteins to nestin and the increased expression of sHSPs in in reactive astrocytes and endothelial cells are caused by pathogenic microorganism induced inflammatory stress. FEBS Lett, 2002 Oct 2, 529(1), 17 - 21 Prion diseases: pathogenesis and public health concerns; Dormont D; Transmissible spongiform encephalopathy (TSE) agents or prions induce neurodegenerative fatal diseases in humans and in some mammalian species . Human TSEs include Creutzfeldt-Jakob disease (CJD), Gerstmann-Straussler-Scheinker syndrome, kuru and fatal familial insomnia . In animals, scrapie in sheep and goats, feline spongiform encephalopathy, transmissible mink encephalopathy, chronic wasting disease in wild ruminants, and bovine spongiform encephalopathy (BSE), which appeared in the UK in the mid-1980s {Wells, G.A.H . et al . (1987) Vet . Rec . 121, 419-420}, belong to the TSE group . Prions have biological and physicochemical characteristics that differ significantly from those of other microorganisms; for example, they are resistant to inactivation processes that are effective against conventional viruses, including those that alter nucleic acid structure or function . Alternatively, infectivity is highly susceptible to procedures that modify protein conformation . Today, the exact nature of prions remains unknown even though it is likely that they consist of protein only . At the biochemical level, TSEs are characterised by the accumulation, within the central nervous system of the infected individual, of an abnormal isoform of a particular protein from the host, the prion protein {Prusiner, S.B . (1982) Science 216, 136-144} . TSEs are transmissible among their species of origin, but they can also cross the species barrier and induce chronic infection and/or disease in other species . Transmissibility has been proven in natural situations such as the outbreak of CJD among patients treated with pituitary-derived hormones and the appearance of BSE that affected UK cattle in the mid-1980s. Eksp Klin Gastroenterol, 2002, (3), 45 - 8, 120 {Results of long-term observation of patients with ulcers associated with Helicobacter pylori after eradication of the microorganism}; Burakov II; The change of approaches to the ulcer treatment with the inclusion of H . pylori-affecting preparations into therapeutic complexes has greatly modified the course of the disease and allowed to reduce the frequency of its relapses in developed countries {1, 2} . However, less optimistic results were obtained in studies carried out in countries with the low social and economic level {3} . This work presents the results of a long-term supervision over patients with stomach and duodenal ulcers associated with the helicobacteriosis infection. Support Care Cancer, 2002 Sep, 10(6), 466 - 73 Epub 2002 Mar 23. Periodontal infection in cancer patients treated with high-dose chemotherapy; Raber-Durlacher JE et al.; The infected and inflamed periodontium can act as a focus for systemic infection in neutropenic cancer patients . The incidence of these oral infections is unknown, but probably underestimated . Periodontal infections can easily be overlooked, primarily because symptoms of gingival inflammation may be minimal and the infection may be located in deeper parts of the periodontium . Assessment of a patient's periodontal condition before the onset of profound neutropenia is critical to the diagnosis and the management of these potentially life-threatening infections . This review article is aimed at informing supportive care providers of recent insights into the pathogenesis of periodontal diseases and the role of subgingival microorganisms, with the emphasis on these infections in cancer patients treated with high-dose chemotherapy . Furthermore, a multidisciplinary approach to the management of periodontal infections and the need for future research is discussed. J Anim Sci, 2002 Sep, 80(9), 2349 - 61 Effects of ventilation regimen on the welfare and performance of lactating ewes in summer; Sevi A et al.; A 6-wk trial was performed with thirty-six lactating Comisana ewes during the summer of 2001 . The animals were divided into three groups of 12, which were designated low (LVR), moderate (MVR), and programmed (PROGR) ventilation regimens . In LVR and MVR rooms, fans provided 10 ventilation cycles of 12.5 and 25 min/h, respectively, whereas in the PROGR room, the fan was programmed to operate at 30 degrees C air temperature and 70% relative humidity . Mean ventilation rates were 33, 66, and 173 m3/h per ewe in LVR, MVR, and PROGR rooms . Air concentrations of microorganisms and dust and of gaseous pollutants were measured twice weekly . Respiration rate and rectal temperature were monitored throughout the trial at 1430 . Behavioral traits of ewes were recorded once per week from 0930 to 1230 . Cell-mediated immune response to phytohemagglutinin at d 3, 20, and 40 and humoral response to chicken egg albumin at d 11, 21, 30, and 40 were determined . At d 37, ewes were injected with 2 IU porcine ACTH/kg body weight(0.75) and subjected to blood sampling for evaluation of cortisol concentrations immediately before and 1, 2, and 4 h after ACTH injection . Milk yield was recorded daily . Individual milk samples were analyzed weekly for composition and renneting parameters and fortnightly for bacteriological characteristics . Averages of temperature-humidity index values were 78.9, 76.8, and 74.5 in LVR, MVR, and PROGR rooms, respectively . The LVR and MVR treatments resulted in higher NH3 and CO2 air concentrations than PROGR treatment (P < 0.05) . The LVR and MVR ewes had higher rectal temperatures than PROGR ewes (P = 0.001) . LVR animals also exhibited higher idling compared to PROGR (P < 0.01) and lower feeding times than MVR (P < 0.05) and PROGR animals (P < 0.01) . Ewes under the LVR treatment displayed significant lower averages of antibody titers and higher plasma cortisol levels than PROGR (P < 0.01) and MVR ewes (P < 0.05) 60 min after ACTH injection . The LVR treatment resulted in lower yields of milk (P < 0.01) and reduced feed efficiency (P < 0.01) than PROGR treatment . Results suggest that a fan-ventilated system, providing ventilation cycles during the warmest hours of the day and the night at a mean ventilation rate of 66 m3/ewe per hour, may sustain the performance and welfare in lactating ewes raised in warm climates during summer . A ventilation regimen, programmed to operate over upper critical air temperature and relative humidity, may be economically unattractive under these conditions. Glob Issues, 1996 Nov, 1(17), 6 - 9 Here to stay . An interview with Dr . David Satcher, Director, U.S . Centers for Disease Control and Prevention; Stilkind J; PIP: This paper presents an interview with Dr . David Satcher of the US Centers for Disease Control and Prevention (CDC) on the issue of infectious and chronic diseases . The problem of new and reemerging infectious diseases around the world, particularly in developing countries, is discussed . While drug resistant malaria and sporadic outbreaks of Ebola in Africa have alarmed health programs, AIDS is the major emerging infectious disease throughout the world . Microorganisms have a way of surviving: they mutate, they adapt, and they emerge . The CDC inverted question marks major partner in the global response to emerging infections is WHO . The CDC has learned lessons from its experience with AIDS and Ebola virus events . A system of domestic surveillance and global prevention should be developed . Poverty, hunger and malnutrition may contribute to the rise of chronic diseases in developing countries . The introduction of richer diets higher in cholesterol in combination with a sedentary lifestyle also will also lead to more chronic diseases among people in developing countries . In the future, smoking will have a greater impact on mortality than will any other factor in these countries . Infectious diseases are much less predictable . Drug resistant microorganisms and urbanization counterbalance the development of vaccines . The U.S . health care system wrongly prioritizes tertiary care while spending only 1% of its health budget on the prevention of diseases . Dr . Satcher would like to see the implementation of programs dealing with human behavior, sanitation, immunization and health education as preventive measures . Afr Health, 1998 Mar, 20(3), 10 - 2 Sexually transmitted diseases in Africa: time for action; Gerbase AC et al.; PIP: Sexually transmitted diseases (STDs) are major public health problems which often lead to serious complications and sequelae, including infertility . Infection with STDs also facilitates the transmission of HIV, making the early diagnosis and care of STDs integrated into other services one of the most cost-effective strategies to prevent the spread of HIV . The direct and indirect costs of STDs worldwide are considerable . Sub-Saharan Africa ranks first in STD yearly incidence compared to other world regions . The World Health Organization has estimated that every year in Africa there are 3.5 million cases of syphilis, 15 million cases of chlamydial disease, 16 million cases of gonorrhea, and 30 million cases of trichomoniasis . STDs are a high public health priority especially because of their widespread prevalence and treatability . Herpes simplex virus infection and human papillomavirus infection are growing problems in sub-Saharan Africa . While STDs are caused by more than 20 microorganisms, they present themselves mainly in 4 syndromes and may therefore be treated syndromically . Africa must implement effective and comprehensive integrated activities against the STD epidemic . Elements of such a strategy will include disease prevention, screening and case finding, and the early diagnosis and treatment of cases . Shengzhi Yu Biyun, 1997, 8(1), 15 - 21 The relationship between infection and the tailed IUD; Jin BH et al.; PIP: To clarify the association between the tailed IUD and pelvic infection, vaginal smears from 111 Chinese IUD users (40 acceptors of tailed IUDs and 71 of tailless devices) were analyzed for bacterial microorganisms . All study subjects had used the IUD for at least 5 years . Of the 11 samples positive for bacteria taken from the internal os, eight were from users of tailed IUDs (p 0.01) . Of the 7 positive samples taken from the uterine cavity, 5 were linked to a tailed device . Although the correlation between the tailed IUD and pelvic symptoms was not significant, infected users of the tailed device were slightly more likely to be symptomatic . The tailed IUD did not increase leukorrhea excretion . ZPG Report . 1991 Dec;23(6):1, 4. Unraveling the tapestry of life . Can we mend our earth? Sherman D. PIP: The industrial revolution has reached its most significant turning point . For the 1st time in its history the limiting factor in its growth has not been our ability to utilize resources, but rather a lack of the resources themselves . At the same time the largest problem to ever face our species, the loss of biodiversity, is threatening our own survival . Clearly we can not support our current numbers under the current consumptive system . All our food is provided by wild species that have been genetically manipulated to serve our ends . Of the 80,000 known edible plant species only about 200 are cultivated regularly and only 3 (corn, wheat, rice) function as staples . Many of our industrial resources come from natural sources . 40% of our pharmaceuticals come from wild sources . It is impossible to isolate ourselves from the Earth ecosystem . Soil microorganisms are but one example of our dependence upon nature . The rainforests also serve as our planets lungs, without them we will choke to death . Yet we continue to destroy them at an alarming rate . They account for only 7% of our land area, and they have already shrunk some 40% . In purely economic terms the wholesale destruction of the rainforests is bad business: in rubber and fruit, 2.5 acres of rainforest is worth $9000 annually compared to cow pasture ($2500) and logging ($1000) . Of course with fruit and rubber the rainforest remains sustainable while cow pasture and logging mean the destruction of the rainforest . Network, 1989, 10(4), 12 - 3 Tarnished copper IUDs still safe and effective. {Contraception and pelvic infection in women} Keith L, Berger GS, Brown ER. PIP: Although sexually transmitted diseases are a major public health problem at the international level, the relationship between contraception and pelvic infection is seldom examined . Numerous STDs are more difficult to diagnose, more frequent, and more serious in women than in men . Differential diagnosis between pelvic infection and other intraabdominal syndromes has been a concern for practitioners for years, and many pelvic infections are probably never diagnosed . Lower abdominal pain and sensitivity as well as fever, leucocytosis, accelerated sedimentation rate, inflammatory annexial mass evident on sonography, and microorganisms in the pouch of Douglass and presence of leucocytes in the peritoneal fluid are diagnostic criteria . Apart from errors in treatment resulting from errors in diagnosis, pelvic infections are often inadequately treated, especially in the initial phase before symptoms are confirmed . The exact incidence of pelvic infections in the US is unknown, but pelvic inflammatory disease (PID) accounted for over 200,000 hospitalizations per year between 1970-75 . PID carries grave risks of subsequent ectopic pregnancy, chronic pelvic pain, and infertility which is more likely as the number of acute episodes increases . The female genital tract has diverse microenvironments propitious for growth of microorganisms of different types, aerobic and anaerobic . Each anatomic site has specific features conditioning bacterial growth . Histological modifications during the menstrual cycle and pregnancy affect the microbial flora . Except in the case of gonorrhea, it is not known how many female lower genital tract infections spread to the upper tract . Since 1970, several studies have domonstrated a growing diversity of cervical and vaginal flora in asymptomatic subjects . The principal risk factors for PID have been well described in the literature . All contraceptive methods except the IUD provide some degree of protection against PID . Even among IUD users the risk of PID is probably not greater than among women with a comparable risk of exposure to STDs . The protective effect of condoms has been recognized since the era of Casanova, but it is difficult to quantify . Studies describing the protective effects of spermicides used one against pelvic infection are very rare, and protective effects have usually been demonstrated only in vitro . Surfactants such as nonoxynol probably have viricidal properties against herpes simplex . Condoms and diaphragms have been seen to exercise a protective effect independent of spermicide, with relative risks of .6 and .4 compared to nonouse of contraception . There is as yet no consensus on changes in risk of PID during oral contraceptive (OC) use, but several studies have shown OCs to have a protective effect . Risks of PID in IUD users apparently stem from contamination during insertion or of the thread during prolonged use, but both possibilities remain controversial . The use of women not using contraception as controls in studies of relative risks of PId may not be appropriate because their sexual behavior and risks of exposure to STDs may differ . At the moment of ovulation, when the mucus is most receptive, IUDs do not place any barrier in the way of ascension of sperm and bacteria to the upper genital tract . Prikl Biokhim Mikrobiol, 2002 Jul-Aug, 38(4), 355 - 73 {Cellulases from microorganisms}; Rabinovich ML et al.; Compositions of cellulase-hemicellulase systems of aerobic fungi (hyphomycetes, ascomycetes, and basidiomycetes), aerobic bacteria, actinomycetes, as well as anaerobic fungi and bacteria, are considered in the context of modern structural classification of glycosyl hydrolases . A new nomenclature of cellulases and relative enzymes based on their structural classification is reviewed . Some opportunities of cellulase improvement by means of protein engineering are discussed. Chembiochem, 2002 Jul 2, 3(7), 619 - 27 Big effects from small changes: possible ways to explore nature's chemical diversity; Bode HB et al.; Fungi or bacteria that produce secondary metabolites often have the potential to bring up various compounds from a single strain . The molecular basis for this well-known observation was confirmed in the last few years by several sequencing projects of different microorganisms . Besides well-known examples about induction of a selected biosynthesis (for example, by high- or low-phosphate cultivation media), no overview about the potential in this field for finding natural products was given . We have investigated the systematic alteration of easily accessible cultivation parameters (for example, media composition, aeration, culture vessel, addition of enzyme inhibitors) in order to increase the number of secondary metabolites available from one microbial source . We termed this way of revealing nature's chemical diversity the 'OSMAC (One Strain-Many Compounds) approach' and by using it we were able to isolate up to 20 different metabolites in yields up to 2.6 g L(-1) from a single organism . These compounds cover nearly all major natural product families, and in some cases the high production titer opens new possibilities for semisynthetic methods to enhance even more the chemical diversity of selected compounds . The OSMAC approach offers a good alternative to industrial high-throughput screening that focuses on the active principle in a distinct bioassay . In consequence, the detection of additional compounds that might be of interest as lead structures in further bioassays is impossible and clearly demonstrates the deficiency of the industrial procedure . Furthermore, our approach seems to be a useful tool to detect those metabolites that are postulated to be the final products of an amazing number of typical secondary metabolite gene clusters identified in several microorganisms . If one assumes a (more or less) defined reservoir of genetic possibilities for several biosynthetic pathways in one strain that is used for a highly flexible production of secondary metabolites depending on the environment, the OSMAC approach might give more insight into the role of secondary metabolism in the microbial community or during the evolution of life itself. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2002 Sep, 94(3), 281 - 93 Endodontic infections: concepts, paradigms, and perspectives; Siqueira JF Jr; Overwhelming evidence indicates that periradicular diseases are infectious disorders . The question now is no longer whether microorganisms are involved in the pathogenesis of such diseases, but which specific microbial species are . The list of microorganisms involved in periradicular diseases keeps expanding and has the potential to become increasingly more accurate during the next few years . Molecular methods have contributed significantly to the knowledge about the microbial species involved . Undoubtedly, a great deal of additional research is needed to define the specific role played by suspected endodontic pathogens in the etiology of each form of periradicular disease and to determine the best therapeutic measures for the pathogen's eradication . In addition, there is an emergent need to define markers that permit the clinician to know when he or she should conclude the treatment and to predict the outcome of the treatment . Although endodontic procedures and some acute endodontic infections can cause bacteremia, there is no clear evidence that microorganisms from the root canal can cause diseases in remote sites of the body . However, there is a risk in some compromised individuals, and prophylactic measures should be taken . Prescription of systemic antibiotics in endodontic therapy is rarely necessary . Because of the emergence of bacterial resistance against most known antibiotics, their use in endodontics should be highly limited and restricted to a few cases. Thorax, 2002 Oct, 57(10), 917 - 8 Barking up the wrong tree? Use of polymerase chain reaction to diagnose syphilitic aortitis; O'Regan AW et al.; The presentation of syphilitic aortitis is often atypical and available serological tests are non-specific . The diagnostic gold standard remains direct identification of microorganisms in tissue . We present a case of syphilitic aortitis that presented as a mediastinal mass and report the use of polymerase chain reaction for Treponema pallidum to diagnose syphilitic aortic disease. Appl Environ Microbiol, 2002 Oct, 68(10), 4827 - 34 Characterization and implications of the cell surface reactivity of Calothrix sp . strain KC97; Phoenix VR et al.; The cell surface reactivity of the cyanobacterium Calothrix sp . strain KC97, an isolate from the Krisuvik hot spring, Iceland, was investigated in terms of its proton binding behavior and charge characteristics by using acid-base titrations, electrophoretic mobility analysis, and transmission electron microscopy . Analysis of titration data with the linear programming optimization method showed that intact filaments were dominated by surface proton binding sites inferred to be carboxyl groups (acid dissociation constants {pK(a)} between 5.0 and 6.2) and amine groups (mean pK(a) of 8.9) . Sheath material isolated by using lysozyme and sodium dodecyl sulfate generated pK(a) spectra similarly dominated by carboxyls (pK(a) of 4.6 to 6.1) and amines (pK(a) of 8.1 to 9.2) . In both intact filaments and isolated sheath material, the lower ligand concentrations at mid-pK(a) values were ascribed to phosphoryl groups . Whole filaments and isolated sheath material displayed total reactive-site densities of 80.3 x 10(-5) and 12.3 x 10(-5) mol/g (dry mass) of cyanobacteria, respectively, implying that much of the surface reactivity of this microorganism is located on the cell wall and not the sheath . This is corroborated by electrophoretic mobility measurements that showed that the sheath has a net neutral charge at mid-pHs . In contrast, unsheathed cells exhibited a stronger negative-charge characteristic . Additionally, transmission electron microscopy analysis of ultrathin sections stained with heavy metals further demonstrated that most of the reactive binding sites are located upon the cell wall . Thus, the cell surface reactivity of Calothrix sp . strain KC97 can be described as a dual layer composed of a highly reactive cell wall enclosed within a poorly reactive sheath. Curr Opin Pharmacol, 2002 Oct, 2(5), 507 - 12 Two-component and phosphorelay signal-transduction systems as therapeutic targets; Stephenson K et al.; Two-component and phosphorelay signal-transduction systems of pathogenic bacteria control the expression of genes encoding virulence factors and essential functions . Recent systematic gene inactivation studies have confirmed the integral role of two-component systems in the pathogenesis of diseases caused by several microorganisms and highlighted the validity of using these systems as targets for therapeutic intervention . Structural studies of signal-transduction proteins have recently revealed common features that may allow rational drug design for therapeutic intervention . In particular, the conserved domains of response regulators may represent the best targets for inhibition. Chem Biol, 2002 Sep, 9(9), 956 - 8 Toward bioengineering anticancer drugs; Kerwin S; The biosynthetic route for enediyne production remained mysterious until two independent groups recently reported the genes that orchestrate enediyne synthesis in two different microorganisms . These discoveries lay the foundations for engineering this pathway to generate improved anticancer drugs. People Planet, 1997, 6(3), 14 - 7 The threatened plague; Epstein P; PIP: This article discusses changes in disease patterns affecting human health that may be related to environmental and social changes in the world . The World Health Report reveals that 30 new diseases emerged in the past 20 years . Old diseases are becoming resistant to new drugs . Infectious diseases that were in decline are spreading: diphtheria, whooping cough, and measles . Illnesses such as malaria, fevers, cholera, and rodent-borne viruses are becoming more frequent . Diseases that are transmitted by animals or water are related to environmental and social changes . Degraded environments are susceptible to the appearance of opportunistic species, such as weeds, rodents, insects, and microorganisms . Stable environments support the welfare of large predators and control opportunistic species . Owls, coyotes, and snakes eat rodents that carry Lyme disease ticks and a variety of viruses, plague, and bacteria . Reptiles, birds, spiders, ladybugs, bats, and fish consume larvae and mosquitoes that cause malaria and fevers . Habitat loss and fragmentation, monocultures, excessive use of toxic chemicals, climate change, and weather instability are widespread global changes that reduce the predator population . Small wilderness habitats favor pests . Monocultures reduce genetic diversity and increase vulnerability . Excessive use of pesticides harms birds and helpful insects . A sign of a failing ecosystem is the population explosion of pests and disequilibrium . The Environmental Distress Syndrome is characterized as: 1) emerging infectious diseases, 2) loss of biodiversity, 3) increased generalist species and decreased specialist species, 4) declines in specific specialists, such as pollinators responsible for preservation of flowering plants, and 5) increased coastal algal blooms . The impacts of disease mean considerable costs to humans, agriculture, and livestock . Loss of resources is also costly . Contracept Technol Update, 1995 May, 16(5), 59 - 61 New label broadens IUD candidacy profile . History of some diseases now just a precaution; Experimental cervical cap has advantages over other methods; PIP: A physician in San Diego, California, developed a new cervical cap, "Fem Cap." A feasibility study (N = 121) showed that, at 1 year, it was 95.2% effective at preventing pregnancy . It became dislodged in 2 of 5 women who conceived, while the other 3 women did not use the cap during every act of intercourse . 8 women experienced dislodgement because they did not check to ensure proper placement over the cervix . Suction caused about 20% of the women to have difficulty removing the cap, but after several months of use, this problem was reduced to 2-3% . None of the sexual partners objected to the cap . The Fem Cap has an upturned brim which fits and adheres to the vaginal walls . The dome is shaped to completely cover the posteriorly longer and anteriorly shorter cervix . The groove between the brim and the done acts as a reservoir for the spermicide, keeping the vaginal wall from absorbing the spermicide, and for the sperm . Fem Cap's material, silicone rubber, is nonporous, nonreactive, and nonallergenic . It does not harbor microorganisms and can be cleaned easily and sterilized . The 24 mm cap is for nulliparous women . The 32 mm cap is for women who have undergone vaginal delivery . Women who cannot be fitted for either of these 2 sizes can almost always be fitted for the 28 mm size . Such women tend to include those who have undergone abortion or Cesarean section . Insertion is very easy . Health care providers can train patients in insertion and removal of Fem Cap in 15-20 minutes . The physician had developed a video on inserting and removing Fem Cap . Giving this video and written instructions to women in the study resulted in 90% learning very well how to use it . The safety and calmness of practicing in the privacy of one's own home contributed to this success . Estimated cost of the Fem Cap is about $60 . Women can use it for about 3 years . Popul Sci, 1987, 7, 55 - 8 Breast feeding as seen by Islam; Hawwas AW; PIP: Islamic law requires mothers to breast feed their children for 2 years . the father must assist the mother in breast feeding the children by providing her with food and clothing . If the father dies or does not live at home, the heir must support the mother thereby allowing her to breast feed her infants . A wet nurse can feed a child only if the mother's milk is not available . Allah recognizes that breast milk is the most wholesome food source for infants . It is made daily based on the infant's needs for growing and nourishment, while other milks and/or formulas remain the same regardless of the infant's daily requirements . For example, the 1st milk, a small amount of yellowish colostrum, contains protein, minerals, and antibodies . Colostrum also acts as a laxative . Breast milk is also comprised of enzymes which make breast milk easy to digest . These digestive enzymes also destroy any microorganisms in the stomach that may have entered thereby providing extra protection against disease . Further, breast milk is sterile . Infant mortality rates (IMRs) among breast fed infants are 4 time lower than IMRs of artificially fed infants . In addition, breast feeding establishes a strong bond between mother and child in which the child feels happiness and security . Breast feeding also benefits the mother . The uterus of a breast feeding mother fully contracts to its original size, while the uterus of a mother who does not breast feed does not . Further, breast feeding women are less prone to breast cancer than those women who have never breast fed . In addition, the more a women breast feeds the less likely she is to get breast cancer . Breast feeding also acts to a certain degree as a contraceptive . Jinko Mondai Kenkyu, 1983 Jan, (165), 48 - 53 {Sex differences in mortality in Japan}; Takahashi S; PIP: An analysis of sex differentials in mortality in Japan is presented . In particular, the author explores differences in causes of death by sex between 1950 and 1980 . It is concluded that the increase in the differences in life expectancy by sex is primarily due to the improvement in mortality of persons over 65 years of age . The significance of structural changes in the causes of death over this time period is also considered, as internal causes became more important than causes related to microorganisms . Biol Zent Bl, 1976 July-August, 95(4), 405 - 13 {Mutagenicity testing with Drosophila as a method for detecting potential carcinogens}; Vogel E et al.; PIP: Drosophila, a higher organism with a short generation period, which can be reproduced quickly, cheaply, and in large numbers, offers, at the present time, the most rapid process for detecting point mutations and small deletions in animals . A technique perfected in the last decades provides subtle methods for detecting all mutation types relevant to humans . Of the different testing procedures, the extremely sensitive Basc-technique is most highly recommended for ascertaining X-chromosomal recessive lethal mutations; forward mutations and small deletions may be demonstrated in 115 of the total genomes . Enzyme systems in Drosophila are similar to liver microsomes in their metabolic capacity and indirect carcinogens can be detected as mutagens . Thus Drosophila procedures can be used to corroborate findings of preliminary tests with microorganisms . A further advantage of Drosophila is the practicable identification of substances with short-lived metabolites as end products of activation processes in the gonads . Biofizika, 2002 Jul-Aug, 47(4), 677 - 82 {Adaptive mutagenesis accelerates microorganism evolution only while competing for resources: mathematic modeling}; Vel'kov VV et al.; Mathematical simulation of adaptive mutagenesis was carried out . The results obtained indicate that adaptive mutagenesis can play an essential role in the survival of microbial populations . We show that the competition for natural resources is a necessary factor of adaptive mutagenesis. Trends Immunol, 2002 Oct, 23(10), 480 - 5 Immune escape through C-type lectins on dendritic cells; Engering A et al.; Dendritic cells (DCs) detect different pathogens and elicit tailored anti-microbial immune responses . They express C-type lectins that recognise carbohydrate profiles on microorganisms, resulting in internalisation, processing and presentation . Intracellular sequences of distinct DC-specific lectins point to differences in intracellular routing that influence antigen presentation . Moreover, putative signalling motifs hint to the activation of DCs on carbohydrate recognition . Recent evidence shows that not only pathogens, but also tumour antigens, exploit C-type lectins to escape intracellular degradation resulting in abortive immunity . More insight into ligand specificity, intracellular targeting and signalling will reveal the pathways by which pathogens modulate immunity through C-type lectins. AIDS Wkly Plus . 1997 Jul 21;:19-20. Controversial report warns of recycled glove use in Kenyan hospital; Some unknown episodes in the history of population dynamics concerning the logistic law: the lesson of the great epidemiologists; The author discusses the work of "the two great epidemiologists R . Ross and A . G . McKendrick . This talk is basically devoted to their inverted question marklesson' about logistic theory and it is organised as follows: section 2 is devoted to McKendrick's {1911} contribution to the growth of microorganism populations, a paper which was not widely acclaimed, but which marks the first experimental laboratory verification of the logistic law . Section 3 is also devoted to McKendrick and to the way he repeatedly derived the logistic equation as a model for inverted question marksimple epidemics' . Finally, in section 4, we present Ross's inverted question markA priori pathometry' theory that really emphasizes...the role played by the logistic law as the inverted question markfirst principle' of the mathematical epidemiology." excerpt Profamilia, 1991 Dec, 7(18), 28 - 30 {Towards an urban world}; The problem of septic abortion in Tanta University hospital: a clinical and bacteriological study; PIP: Clinical and bacteriological features of 141 cases of septic abortion admitted to Tanta University Hospital (Egypt) are reviewed . The pathogenesis and management of septic abortion, septic shock, and acute renal failure are discussed, and the types of microorganisms identified in uterine and cervical cultures are tabulated . Itiis recommended that bacteriological examination should routinely be performed in all cases of septic abortion in order to identify the micoorganisms involved so that appropriate measures may be taken to prevent serious complications . Jinko Mondai Kenkyu, 1982 Oct, (164), 19 - 36 {Causes of death and age patterns of mortality in Japan since 1950}; Takahashi S; PIP: Trends in mortality in Japan since 1950 are analyzed . The relationship between changes in the structure of causes of death and the level of overall mortality is examined using a variety of demographic techniques . Changes in the crude death rate, the age-standardized death rate, and life expectancy at birth are reviewed, and variations in mortality resulting from changes in the age structure of the population are considered . Changes in the causes of death by sex are also examined over time . It is noted that between 1950 and 1965, major reductions were made in mortality from diseases caused by microorganisms; between 1965 and 1980, the major reductions were in mortality from cerebrovascular diseases and other diseases not caused by microorganisms . Wien Med Wochenschr, 2002, 152(15-16), 379 - 81 {Acid oligosaccharides as the active principle of aqueous carrot extracts for prevention and therapy of gastrointestinal infections}; Kastner U et al.; Adherence of microorganisms to the intestinal mucosa is an important and initial step in the pathogenesis of gastrointestinal infections and mediated by carbohydrate structures on the cell surface . Adherence can be blocked by carbohydrate receptor analogues . Aqueous extracts from carrots (carrot soup) contain acidic oligosaccharides, which are able to block adherence of various enteropathogenic microorganisms to HEp-2 cells and human intestinal mucosa in vitro . Dependent on the grade of polymerisation the most potent blocking ability was seen for trigalacturonic acid . Clinical studies revealed, that aqueous carrot extracts are significantly superior to the basic glucose-electrolyt-solution for oral rehydration in acute gastrointestional infections of children. Proc Natl Acad Sci U S A, 2002 Oct 1, 99(20), 13043 - 8 Epub 2002 Sep 20. C-reactive protein binds to both oxidized LDL and apoptotic cells through recognition of a common ligand: Phosphorylcholine of oxidized phospholipids; Chang MK et al.; C-reactive protein (CRP) is an acute-phase protein that binds specifically to phosphorylcholine (PC) as a component of microbial capsular polysaccharide and participates in the innate immune response against microorganisms . CRP elevation also is a major risk factor for cardiovascular disease . We previously demonstrated that EO6, an antioxidized LDL autoantibody, was a T15 clono-specific anti-PC antibody and specifically binds to PC on oxidized phosphatidylcholine (PtC) but not on native PtC . Similarly, EO6 binds apoptotic cells but not viable cells . In addition, such oxidized phospholipids are recognized by macrophage scavenger receptors, implying that these innate immune responses participate in the clearance because of their proinflammatory properties . We now report that CRP binds to oxidized LDL (OxLDL) and oxidized PtC (OxPtC), but does not bind to native, nonoxidized LDL nor to nonoxidized PtC, and its binding is mediated through the recognition of a PC moiety . Reciprocally, CRP binds to PC, which can be competed for by OxLDL and OxPtC but not by native LDL, nonoxidized PtC, or by oxidized phospholipids without the PC headgroup . CRP also binds to apoptotic cells, and this binding is competed for by OxLDL, OxPtC, and PC . These data suggest that CRP binds OxLDL and apoptotic cells by recognition of a PC moiety that becomes accessible as a result of oxidation of PtC molecule . We propose that, analogous to EO6 and scavenger receptors, CRP is a part of the innate immune response to oxidized PC-bearing phospholipids within OxLDL and on the plasma membranes of apoptotic cells. Scand J Infect Dis, 2002, 34(8), 628 - 9 Bordetella bronchiseptica pleural infection in a patient with AIDS; Viejo G et al.; Bordetella bronchiseptica is an important veterinary pathogen but a rare cause of opportunistic respiratory tract infection in humans . We describe a case of pleural effusion caused by B . bronchiseptica in a patient with AIDS who was coinfected with Mycobacterium tuberculosis . To the best of our knowledge this is the first case in the medical literature in which the microorganism has been isolated from pleural fluid. Inflammation, 2002 Oct, 26(5), 215 - 22 Dexamethasone inhibits apoptosis of human neutrophils induced by reactive oxygen species; Ruiz LM et al.; Neutrophils are completely differentiated cells that die in tissues a few days after they migrate from the vascular compartment as a consequence of a rigouous apoptotic program . Many of the mediators produced during an inflammatory response delay neutrophil apoptosis allowing a more efficient removal of microorganisms but also favoring the tissue damage by reactive oxygen species (ROS) and lysosomal proteins released by neutrophils . Glucocorticoids delay the apoptosis of neutrophils but the mechanisms are not completely understood . To investigate the inhibition of glucocorticoids on neutrophil apoptosis we have used the glucose/glucose oxidase (G/GO) system as a constant source of hydrogen peroxide . When neutrophils are incubated in the presence of the G/GO system, a significant acceleration of their apoptotic response is observed . Preincubation with 10(-6) M, 10(-7) M, 10(-8) M or 10(-9) M of dexamethasone, negatively modulated the spontaneous and G/GO induced apoptosis of neutrophils . Then the G/GO system is a useful model to simulate the oxidative stress of neutrophils, and that the effect of DXM on neutrophil apoptosis depends, at least in part, on blocking the proapoptotic effect of ROS. Rocz Panstw Zakl Hig, 2002, 53(2), 197 - 202 {Tests of the microbiologic barrier in sterile packaging}; Jakimiak B et al.; After decontamination, cleaning, maintenance and functional testing, sterilised items must be packed suitably . The package must protect sterilised items against microbial contamination during removal from the sterilising chamber, and during storage or transport until use . The ability of any given pack to withstand penetration by bacteria is termed bacterial barrier efficiency . The assurance to keep the contents of the packaging sterile is determined by quality of the materials and the quality of seals . The research was done to proof the microbial barrier, determined by quality of the packaging materials/systems used to sterilization in hospital practice in Poland . 4 packaging materials and 4 types of containers were tested . In all used methods the growth of microorganisms indicated the lack of the bacterial efficiency, the lack of growth--the bacterial barrier efficiency . It was proved that two tested specialistic sterilization papers, non woven material and specialistic containers (metal with a filter or valve, plastic with a filter) are effective microbial barriers because it was no microorganisms growth . It has been determined that sulphite bleachery paper of 60 g/m2 and Schimmelbusch container do not provide the microbial barrier . Based on the performed studies it has been determined that sulphite bleachery paper and Schimmelbusch container can not be used as sterilization packaging system in sterilization practice in hospitals. Arch Med Res, 2002 Jul-Aug, 33(4), 398 - 404 From noxiustoxin to scorpine and possible transgenic mosquitoes resistant to malaria; Possani LD et al.; Scorpion venom contains different types of peptides toxic to a variety of organisms whose molecular targets have been described as mainly ion-channels of excitable cells where they cause impairment of function . Based on mouse, cricket, and crustacean bioassays, specific toxins for each group of animals have been found . Chromatographic techniques were used to isolate and chemically characterize these peptides . One of the best-studied peptides is noxiustoxin, a 39-amino acid residue-long peptide specific for K(+)-channels . Hadrurin is another scorpion venom peptide whose activity was shown to be bactericidal to a variety of species . Structural similarities of a newly discovered peptide (scorpine) with those of defensins and cecropins showed that scorpion venom contains peptides toxic to microorganisms and malaria parasites . Scorpine was shown to disrupt the sporogonic development of Plasmodium berghei . Using this system as a model for malaria, we introduced the gene of scorpine into a vector for generation of transgenic flies resistant to the infection by Plasmodium . The final aim of this work is to incorporate this gene under the promoter of proteolytic enzymes of digestive tract of mosquitoes for synthesis and liberation of toxic peptide(s) into stomach of freshly fed mosquitoes potentially carrying Plasmodium gametes . In this manner, a putative transgenic mosquito with these characteristics would secrete a toxic peptide with digestive enzymes into midgut, impairing proper development of Plasmodium, hence controlling malaria, one of the most important tropical diseases worldwide. J Rheumatol, 2002 Sep, 29(9), 1942 - 9 Uricase formulated with polyethylene glycol (uricase-PEG 20): biochemical rationale and preclinical studies; Bomalaski JS et al.; OBJECTIVE: Humans have a non-sense codon inserted into the 5 prime end of the open reading frame of urate oxidase, and thus express an enzymatically inactive fragment of this enzyme; and consequently are unable to metabolize uric acid into allantoin and are prone to develop hyperuricemia and gout . Various urate oxidases (uricase) from mammals and microorganisms have been administered to humans with hyperuricemia and gout . Although successful in lowering plasma uric acid, these therapies have had limited application due to undesirable biochemical properties of the enzymes used, the short circulating half-life, and inherent antigenicity of these preparations . METHODS: We compared urate oxidase from a variety of sources for specific enzyme activity, pH optimum, affinity, and retention of enzyme activity under physiological conditions . A variety of polyethylene glycols (PEG) were tested to formulate uricase . RESULTS: Urate oxidase from Candida utilis had more favorable enzymatic properties and PEG of 20,000 MW (termed uricase-PEG 20) had greatly reduced antigenicity and increased circulating half-life as compared to those previously described . CONCLUSION: It is anticipated that uricase-PEG 20 may have utility as a treatment for hyperuricemia and gout. J Food Prot, 2002 Sep, 65(9), 1480 - 2 Use of pulsed ultraviolet laser light for the cold pasteurization of bovine milk; Smith WL et al.; Because of concerns that some potentially dangerous microorganisms may survive conventional heat pasteurization of milk and because the heat needed to sterilize milk affects marketability, the ability to efficiently cold pasteurized milk may become more desirable . In this pilot study, we investigated the use of pulsed ultraviolet (PUV) laser light to nonthermally (cold) pasteurized bovine milk . Dairy bulk tank milk was treated with UV light (248 nm) emitted from a pulsed excimer laser . The samples were then analyzed for surviving bacteria by spiral plate counting and subculturing in Trypticase soy broth . Other bulk tank milk samples were inoculated with one of eight relevant milk bacterial species before being exposed to laser light . There was no growth observed for any of the plated or subcultured samples exposed to 25 J/cm2 . One bacterial isolate was then used to inoculate milk to further investigate bactericidal laser light doses . Growth was observed for samples treated with an average of 0.3 to 6.6 J/cm2 but not for those treated with 12.6 J/cm2 . The results indicate that in principle, the bacterial content of milk can be adequately controlled by exposure to PUV laser light. Plant Physiol, 1994 Sep, 106(1), 71 - 77 Iron Inefficiency in Maize Mutant ys1 (Zea mays L . cv Yellow-Stripe) Is Caused by a Defect in Uptake of Iron Phytosiderophores; Von Wiren N et al.; To determine the Fe inefficiency factors in the maize mutant ys1 (Zea mays L . cv Yellow Stripe), root exudates of Fe-inefficient ys1 and of two Fe-efficient maize cultivars (Alice, WF9) were collected in axenic nutrient solution cultures . Analysis by thin-layer chromatography and high-performance liquid chromatography revealed that under Fe deficiency ys1 released the phytosiderophore 2{prime}-deoxymugineic acid (DMA) in quantities similar to those of Alice and WF9 . Under nonaxenic conditions, DMA released by plants of all three cultivars was rapidly decomposed by microorganisms in the nutrient solution . Uptake experiments with 59Fe-labeled DMA, purified from root exudates of either Fe-deficient Alice or ys1 plants, showed up to 20 times lower uptake and translocation of 59Fe in ys1 than in Alice or WF9 plants . The presence of microorganisms during preculture and short-term uptake experiments had no significant effect on uptake and translocation rates of 59Fe in Alice and ys1 plants . We conclude that Fe inefficiency in the maize mutant ys1 is the result of a defect in the uptake system for Fe-phytosiderophores. Exp Mol Pathol, 2002 Oct, 73(2), 128 - 34 Ultrastructure of the interaction between mycobacterium tuberculosis- H37Rv-containing phagosomes and the lysosomal compartment in human alveolar macrophages; Borelli V et al.; The ultrastructure of the interaction between the lysosomal compartment and the Mycobacterium tuberculosis-containing phagosomes in human resident alveolar macrophages has been analyzed in detail . Our findings confirm the widely accepted notion that the parasitophore vacuole is made nonfusogenic by the microorganism; however, the association between the lysosomal compartment and the phagosomes does not seem to be impaired as the organelles were shown to spread around the ingested pathogen . Furthermore, interruptions in the phagosome membrane that connect the bacterial surface with the cytosol were observed. J Inorg Biochem, 2002 Sep 30, 92(1), 37 - 42 Bactericidal activity of Ag-zeolite mediated by reactive oxygen species under aerated conditions; Inoue Y et al.; The bactericidal activity induced by the introduction of silver ions into zeolite was studied . Escherichia coli was used as the test microorganism . Silver ions were loaded into zeolite by the ion-exchange method . Silver-loaded zeolite was demonstrated the strong bactericidal activity . Dissolved oxygen was an essential factor for the occurrence of the bactericidal activity because the activity was observed only under aerated condition . Superoxide anions, hydrogen peroxide, hydroxyl radicals and singlet oxygen were formed . Scavengers of these each reactive oxygen species (ROS) inhibited the bactericidal activity . This means that all ROS contributed to the activity. Bioresour Technol, 2002 Nov, 85(2), 107 - 11 Composting of a crop residue through treatment with microorganisms and subsequent vermicomposting; Singh A et al.; Preliminary studies were conducted on wheat straw to test the technical viability of an integrated system of composting, with bioinoculants and subsequent vermicomposting, to overcome the problem of lignocellulosic waste degradation, especially during the winter season . Wheat straw was pre-decomposed for 40 days by inoculating it with Pleurotus sajor-caju, Trichoderma harzianum, Aspergillus niger and Azotobacter chroococcum in different combinations . This was followed by vermicomposting for 30 days . Chemical analysis of the samples showed a significant decrease in cellulose, hemicellulose and lignin contents during pre-decomposition and vermicomposting . The N, P, K content increased significantly during pre-decomposition with bioinoculants . The best quality compost, based on chemical analysis, was prepared where the substrate was treated with all the four bioinoculants together followed by vermicomposting . Results indicated that the combination of both the systems reduced the overall time required for composting and accelerated the composting of ligno-cellulosic waste during the winter season besides producing a nutrient-enriched compost product. Appl Microbiol Biotechnol, 2002 Sep, 59(6), 706 - 12 Epub 2002 Aug 13. Monitoring of morphological development of the arachidonic-acid-producing filamentous microorganism Mortierella alpina; Park EY et al.; Morphological parameters, such as hyphal growth rate, tip formation rate, tip extension rate and branch formation rate, of Mortierella alpina have been measured using a flow-through chamber under 25 different combinations of carbon and nitrogen concentrations . Morphological parameters were influenced not by C/N ratio but by carbon concentration in the medium . Specific rates of hyphal growth and tip formation both remained constant at a low carbon concentration of 5 g/l . Tip extension rate from one tip was 60 microm tip(-1) h(-1) at a carbon concentration below 15 g/l, and the branching formation rate was independent of carbon concentration . Tip extension rate was a function of specific hyphal growth rate, which in turn was linearly proportional to the specific tip formation rate, demonstrating that tip extension rate was exponentially proportional to the specific tip formation rate . Branch formation rate per hyphal element remained unchanged even at tip extension rates lower than 60 microm tip(-1) h(-1) and at specific hyphal growth rates lower than 0.83 h(-1), but decreased drastically at higher rates of tip extension and hyphal growth. Heart Vessels, 2002 May, 16(4), 137 - 45 Chlamydia pneumoniae seropositivity predicts the risk of restenosis after percutaneous transluminal coronary angioplasty; Hayashida K et al.; This study was done to evaluate whether anti-Chlamydia pneumoniae seropositivity can be a predictor of restenosis after coronary intervention . Recent studies indicate that latent infection with C . pneumoniae is associated with and could possibly cause atherosclerosis . However, it is unknown whether chronic infection with this microorganism is involved in the mechanism of restenosis after percutaneous transluminal coronary angioplasty . We prospectively studied 78 consecutive patients (90 target lesions) with symptomatic coronary artery disease who underwent successful coronary intervention to a de novo lesion (conventional balloon angioplasty to 31 lesions and stent implantation to 59 lesions) . At angioplasty, blood samples were collected to measure the serum level of anti-C . pneumoniae IgG to examine whether seropositive patients were prone to restenosis and whether the seropositivity could predict the risk of restenosis determined by follow-up coronary angiography performed within 6 months after the angioplasty . Restenosis, defined as more than 50% stenosis with an increase of 15% or more in the degree of stenosis from that measured on cineangiograms after angioplasty, developed in 36 of 62 seropositive patients and in 4 of 16 seronegative patients (58% vs 25%, P = 0.025) . Lesions in the seropositive patients had a greater mean loss index (mean +/- SD 0.75 +/- 0.45 vs 0.35 +/- 0.41, P < 0.001), which was defined as late loss (luminal diameter reduction at follow-up angiography) divided by acute gain (luminal diameter gain by angioplasty), in late loss (1.07 +/- 0.64mm vs 0.65 +/- 0.79mm, P = 0.019), in percentage of diameter stenosis (57% +/- 20% vs 41% +/- 21%, P = 0.003) and a lesser mean in minimal luminal diameter (1.18 +/- 0.58 mm vs 1.67 +/- 0.63 mm, P = 0.002) at follow-up angiography . In a multivariate logistic regression model, anti-C . pneumoniae IgG seropositivity was a strong independent predictor of restenosis compared to the other risk factors (odds ratio = 6.2, P = 0.01) . C . pneumoniae could play an important role in the mechanism of restenosis and evaluation of the IgG seropositivity, and may help to identify patients at high risk for restenosis. J Microbiol Methods, 2002 Nov, 51(3), 255 - 71 Identification of medically relevant microorganisms by vibrational spectroscopy; Maquelin K et al.; In the recent years, vibrational spectroscopies (infrared and Raman spectroscopy) have been developed for all sorts of analyses in microbiology . Important features of these methods are the relative ease with which measurements can be performed . Furthermore, in order to obtain infrared or Raman spectra, there is only a limited amount of sample handling involved without the need for expensive chemicals, labels or dyes . Here, we review the potential application of vibrational spectroscopies for the use in medical microbiology . After describing some of the basics of the techniques, considerations on reproducibility and standardisation are presented . Finally, the use of infrared and Raman spectroscopy for the (rapid) identification of medically relevant microorganisms is discussed . It can be concluded that vibrational spectroscopies show high potential as novel methods in medical microbiology . Chemosphere, 2002 Sep, 48(9), 975 - 9 Volatile organic compounds from leaves litter; Isidorov V et al.; Qualitative composition of volatile emissions of litter of five species of deciduous trees was investigated by GC-MS . The list of identified substances contains more than 70 organic compounds of various classes . It was established that the composition of components emitted by the litter into the gas phase greatly differs from that of essential oils extracted by hydrodistillation from turned leaves collected from trees during fall . It is suggested that most compounds found in litter emissions are products of vital activity of microorganisms decomposing it . The reported data indicate that after the vegetative period is over the decomposition processes of litter are important seasonal sources of reactive organic compounds under the forest canopy. Int J Food Microbiol, 2002 Sep 15, 78(1-2), 171 - 80 Development and application of new nucleic acid-based technologies for microbial community analyses in foods; Rudi K et al.; Several challenges still persist in the analysis of microorganisms in foods, particularly in studies of complex communities . Nucleic acid-based methods are promising tools in addressing new questions concerning microbial communities . We have developed several new methods in the field of nucleic acid-based microbial community analyses . These methods cover both sample preparation and detection approaches . The sample preparation method involves simplified DNA purification using paramagnetic beads . As an extension of this method, the same paramagnetic beads are used for both cell separation and DNA purification . This enables full automation . The separate detection of viable and dead bacteria is a major issue in nucleic acid-based diagnostics . We have applied a living/dead dye that binds covalently to DNA and inhibits the PCR from dead cells . In addition, a DNA array-based detection assay has been developed . The assay combines the specificity obtained by enzymatic labeling of DNA probes with the possibility of detecting several targets simultaneously by DNA array hybridization . In combination with 16S rDNA amplification, this is a promising tool for community analyses . Also, we have developed a novel approach for multiplex quantitative PCR . The multiplex PCR has been combined with our DNA array-based detection method . Finally, we are now in the process of adapting a system for monitoring microbial growth and death in real-time through the tagging of bacteria with green fluorescent protein (GFP) combined with fluorescence detection using a high-resolution confocal laser scanner. Ying Yong Sheng Tai Xue Bao, 2002 Apr, 13(4), 425 - 8 {Characteristics of functional and nutritious soilless culture substrate for vegetables}; Zhu S et al.; A functional and nutritious substrate for soilless culture, which consists of peanut shell, sawdust, vermiculite, chicken manure, coal cinder, etc, was used to grow cucumbers, tomatoes and peppers in this experiment . The results showed that the substrate was rich in organic matter, N, available P and K, in which the nutrients were basically in balance for three vegetables . High content of microorganisms and high activity of soil enzymes were propitious to the transformation of organic components in the substrate . The yields of cucumber, tomato and pepper increased by 23.83%, 27.34% and 32.98%, respectively . The production value of peppers increased by 180.85%, and its net income increased by 109.69% . The qualities of three vegetables were coincident with 'harmless vegetable' standards. Proc Natl Acad Sci U S A, 2002 Sep 17, 99(19), 12489 - 94 Epub 2002 Sep 09. Folate synthesis in plants: the first step of the pterin branch is mediated by a unique bimodular GTP cyclohydrolase I; Basset G et al.; GTP cyclohydrolase I (GCHI) mediates the first and committing step of the pterin branch of the folate-synthesis pathway . In microorganisms and mammals, GCHI is a homodecamer of approximately 26-kDa subunits . Genomic approaches identified tomato and Arabidopsis cDNAs specifying approximately 50-kDa proteins containing two GCHI-like domains in tandem and indicated that such bimodular proteins occur in other plants . Neither domain of these proteins has a full set of the residues involved in substrate binding and catalysis in other GCHIs . The tomato and Arabidopsis cDNAs nevertheless encode functional enzymes, as shown by complementation of a yeast fol2 mutant and by assaying GCHI activity in extracts of complemented yeast cells . Neither domain expressed separately had GCHI activity . Recombinant tomato GCHI formed dihydroneopterin triphosphate as reaction product, as do other GCHIs, but unlike these enzymes it did not show cooperative behavior and was inhibited by its substrate . Denaturing gel electrophoresis verified that the bimodular GCHI polypeptide is not cleaved in vivo into its component domains, and size-exclusion chromatography indicated that the active enzyme is a dimer . The deduced tomato and Arabidopsis GCHI polypeptides lack overt targeting sequences and thus are presumably cytosolic, in contrast to other plant folate-synthesis enzymes, which are mitochondrial proteins with typical signal peptides . GCHI mRNA and protein are strongly in expressed unripe tomato fruits, implying that fruit folate is made in situ rather than imported . As ripening advances, GCHI expression declines sharply, and folate content drops, suggesting that folate synthesis fails to keep pace with turnover. J Neurol Sci, 2002 Oct 15, 202(1-2), 13 - 23 The role of macrophage/microglia and astrocytes in the pathogenesis of three neurologic disorders: HIV-associated dementia, Alzheimer disease, and multiple sclerosis; Minagar A et al.; Macrophage/microglia (M phi) are the principal immune cells in the central nervous system (CNS) concomitant with inflammatory brain disease and play a significant role in the host defense against invading microorganisms . Astrocytes, as a significant component of the blood-brain barrier, behave as one of the immune effector cells in the CNS as well . However, both cell types may play a dual role, amplifying the effects of inflammation and mediating cellular damage as well as protecting the CNS . Interactions of the immune system, M phi, and astrocytes result in altered production of neurotoxins and neurotrophins by these cells . These effects alter the neuronal structure and function during pathogenesis of HIV-1-associated dementia (HAD), Alzheimer disease (AD), and multiple sclerosis (MS) . HAD primarily involves subcortical gray matter, and both HAD and MS affect sub-cortical white matter . AD is a cortical disease . The process of M phi and astrocytes activation leading to neurotoxicity share similarities among the three diseases . Human Immunodeficiency Virus (HIV)-1-infected M phi are involved in the pathogenesis of HAD and produce toxic molecules including cytokines, chemokines, and nitric oxide (NO) . In AD, M phis produce these molecules and are activated by beta-amyloid proteins and related oligopeptides . Demyelination in MS involves M phi that become lipid laden, spurred by several possible antigens . In these three diseases, cytokine/chemokine communications between M phi and astrocytes occur and are involved in the balance of protective and destructive actions by these cells . This review describes the role of M phi and astrocytes in the pathogenesis of these three progressive neurological diseases, examining both beneficent and deleterious effects in each disease . Arch Otolaryngol Head Neck Surg, 2002 Sep, 128(9), 1061 - 6 Differing pathways of lower airway colonization and infection according to mode of ventilation (endotracheal vs tracheotomy); Morar P et al.; OBJECTIVES: To determine whether the pathogenesis of lower airway colonization and infection was endogenous (via the oropharynx) or exogenous (via the endotracheal tube or tracheotomy) during the 2 modes of ventilation in the same subset of children requiring long-term ventilation . DESIGN: Prospective, observational cohort study . SETTING: A pediatric intensive care unit and a respiratory ward . PATIENTS: Consecutive admissions between September 1, 1993, and August 30, 1998 . MEASUREMENTS AND MAIN RESULTS: Cultures were obtained simultaneously from the oropharynx and tracheobronchial tree on admission to the pediatric intensive care unit, at placement of the tracheotomy, and afterward twice weekly . Forty-five patients were studied . Lower airways were always sterile in 6 children, 39 children (87%) developed a total of 82 episodes of colonization, and 17 (38%) progressed to 25 episodes of infection . The number of infected children was halved once they had a tracheotomy (7 children {16%}) . Of the 107 episodes of colonization and infection, 41 and 66 occurred during endotracheal ventilation and via a tracheotomy, respectively . Primary endogenous episodes of colonization and infection due to bacteria present in the admission flora in the pediatric intensive care unit were significantly more common with endotracheal ventilation than during ventilation via a tracheotomy (31/41 {76%} vs 36/66 {55%}; P =.03) . Secondary endogenous and exogenous episodes of colonization and infection due to bacteria associated with the respiratory ward were significantly more frequent when ventilation was continued through a tracheotomy than during endotracheal ventilation (30/66 {45%} vs 10/41 {24%}; P =.02) . CONCLUSIONS: Surveillance samples allow the distinction between primary endogenous ("imported" bacteria) from secondary endogenous and exogenous ("nosocomial" microorganisms) colonization and infection . This classification permits the development of preventive strategies to control both endogenous and exogenous pathways. Eur J Immunol, 2002 Sep, 32(9), 2460 - 70 Role of chlamydial heat shock protein 60 in the stimulation of innate immune cells by Chlamydia pneumoniae; Costa CP et al.; Chlamydia pneumoniae stimulates potently maturation of and cytokine secretion by bone marrow-derived dendritic cells (BMDDC) . BMDDC responses depend mainly on Toll-like receptor (TLR)2 and to a minor extent on TLR4 . We demonstrate here using C . pneumoniae in an infectious model with the replication-permissive epithelial cell line HEp2 that HSP60 is produced in substantial amounts in chlamydial inclusions during infection . Electron microscopy of chlamydial inclusions revealed that HSP60 was mainly associated with reticulate bodies, but was also located in between the different chlamydial developmental forms . Supernatants of permissive HEp2 cells infected with C . pneumoniae contained soluble chlamydial HSP60 as demonstrated by Western blotting and were able to stimulate BMDDC of wild-type mice . The stimulatory capacity of culture supernatants correlated with the presence of chlamydial HSP60 . In contrast, BMDDC from TLR4-mutant mice crossed to TLR2-deficient mice were not stimulated by the culture supernatant, indicating that chlamydial HSP60 but not cytokines, possibly secreted by infected HEp2 cells, are responsible for the observed stimulation of BMDDC . Purified recombinant HSP60 from C . pneumoniae stimulated BMDDC in a TLR2- and TLR4-dependent fashion similar to the whole microorganism . In summary, these data suggest chlamydial HSP60 as an important mediator of inflammatory responses during infection with C . pneumoniae. Dev Comp Immunol, 2002 Oct, 26(8), 715 - 21 Characterization and partial purification of a lectin from the hemolymph of the white shrimp Litopenaeus schmitti; Cominetti MR et al.; The agglutinating activity of the hemolymph of Litopenaeus schmitti is insensitive to calcium and specific for acetylated sugars, particularly sialic acid (Neu5Ac) and O-sialoglycoconjugates (bovine submaxillary mucin) and has varying specificity for different LPS, which may suggest a putative role in microorganism recognition . Affinity chromatography on fetuin-agarose of the agglutinin resulted in a 220 kDa band (lectin), and a 82.5 kDa band, which probably is hemocyanin . The 220 kDa protein consists of 31 and 34 kDa subunits, suggesting that this lectin is multimeric . The lectin molecular mass was estimated by gel filtration to be 153+/-10 kDa . The hemolymph of L . schmitti comprises at least another soluble lectin, with distinct chemical and carbohydrate specificity than the 220 kDa lectin. J Immunol, 2002 Sep 15, 169(6), 2971 - 8 Regeneration of natural antibody repertoire after massive ablation of lymphoid system: robust selection mechanisms preserve antigen binding specificities; Nobrega A et al.; Natural Abs (NAbs) are Igs present in the serum and body fluids of healthy vertebrate animals, without any previous intentional immunization . NAbs often exhibit autoreactivity but also play an essential role in immunity, being a first line of defense against infectious microorganisms . We have previously analyzed the natural serum IgM Ab repertoire of normal mice, characterizing their reactivity with hundreds/thousands of self Ags; a significant similarity among different individuals was observed, and it was found that many reactivities of NAbs stably kept during adulthood were established early in life, implicating that period as a critical time window in the physiology of NAb repertoire selection . In the work reported here, experiments were conducted to address the role of normal lymphocyte ontogeny to the formation and stability of adult NAb repertoire . The massive destruction of the lymphoid system was promoted in adult mice with gamma-irradiation, and regeneration of hemopoietic tissues was granted by bone marrow or fetal liver inoculum . NAb repertoire regeneration was followed for 60 days after gamma-irradiation in bone marrow or fetal liver chimeric animals . The analysis of serum IgM reactivity with hundreds/thousands of self Ags showed that the NAb repertoire regenerated most of its original format after massive destruction of lymphoid compartments, characterizing autoreactive repertoire selection as a robust biological process . The data also show that regeneration of the NAb repertoire occurred similarly in fetal liver and bone marrow chimeras, although the latter animals poorly reconstituted their CD5(+) B1 cell compartment, suggesting that B1 cells are not essential for natural Ab regeneration. Trends Microbiol, 2002 Sep, 10(9), 425 - 61 Dendritic cells and the complexity of microbial infection; Rescigno M; Dendritic cells (DCs) can discriminate between different classes of microorganisms, present antigens to T cells and initiate innate and adaptive immune responses . DCs do not act individually, and their function can be fine-tuned by environmental and tissue factors as well as by the microorganisms themselves . Recent studies have reported DCs can integrate stimuli derived from microbial pathogens and other cells present at, or recruited to, the site of infection . These interactions can determine the success or failure of the immune response induced against pathogens. Trends Microbiol, 2002 Sep, 10(9), 410 - 8 Nutrient dynamics in the deep blue sea; Karl DM; For more than a century, oceanographers have studied the interactions between the photosynthetic production of organic matter and nutrient dynamics in the sea . This research has been field-oriented and transdisciplinary, occurring at the intersections of research in microbiology, physics, analytical chemistry, cell physiology and ecology . The global database derived from this collective effort established a sound scientific understanding of nutrient dynamics and the vital role of microorganisms, both autotrophic and heterotrophic, in the coupled organic-matter production and decomposition cycles in the sea . However, novel approaches used over the past two decades, including new designs for field experiments, repeat field observations and remote-sensing capabilities, together with updated methods of sample analysis, have led to a revolution in our thinking about the mechanisms and controls of nutrient dynamics in the deep blue sea . Contemporary paradigms bear only partial resemblance to the dogma of the past, and are likely to evolve further as new data and new ideas are presented for open discussion and debate. Naturwissenschaften, 2002 Jul, 89(7), 319 - 21 Plant development in the absence of epiphytic microorganisms; Kutschera U et al.; Microorganisms (bacteria, fungi) are common residents of the roots, stems and leaves of higher plants . In order to explore the dependency of plant development on the presence of epiphytic microorganisms, the achenes (seeds) of sunflower (Helianthus annuus L.) were sterilized and germinated under aseptic conditions . The sterility of the seedlings was determined with the agar impression method . In seedlings from non-sterile seeds (control) that were likewise raised in a germ-free environment, all plant organs investigated (stem, cotyledons and primary leaves) were contaminated with bacteria . Hypocotyl elongation was not affected by epiphytic microorganisms . However, the growth rates of the cotyledons and primary leaves were higher in sterile seedlings compared with the control . The implications of this differential inhibition of organ development by epiphytic bacteria that are transmitted via the outer surface of the seed coat are discussed . We conclude that epiphytes in the above-ground phytosphere are not necessary for the development of the sunflower seedling. Lipids, 2002 Jul, 37(7), 653 - 62 Distinction between esterases and lipases: a kinetic study with vinyl esters and TAG; Chahinian H et al.; The better to characterize enzymes hydrolyzing carboxyl ester bonds (carboxyl ester hydrolases), we have compared the kinetic behavior of various lipases and esterases against solutions and emulsions of vinyl esters and TAG . Short-chain vinyl esters are hydrolyzed at comparable rates by esterases and lipases and have higher limits of solubility in water than corresponding TAG . Therefore, they are suited to study the influence of the physical state of the substrate on carboxyl ester hydrolase activity within a large concentration range . Enzymes used in this study are TAG lipases from microorganisms, lipases from human and guinea pig pancreas, pig liver esterase, and acetylcholinesterase . This study also includes cutinase, a fungal enzyme that displays functional properties between esterases and lipases . Esterases display maximal activity against solutions of short-chain vinyl esters (vinyl acetate, vinyl propionate, and vinyl butyrate) and TAG (triacetin, tripropionin, and tributyrin) . Half-maximal activity is reached at ester concentrations far below the solubility limit . The transition from solution to emulsion at substrate concentrations exceeding the solubility limit has no effect on esterase activity . Lipases are active on solutions of short-chain vinyl esters and TAG but, in contrast to esterases, they all display maximal activity against emulsified substrates and half-maximal activity is reached at substrate concentrations near the solubility limit of the esters . The kinetics of hydrolysis of soluble substrates by lipases are either hyperbolic or deviate from the Michaelis-Menten model and show no or weak interfacial activation . The presence of molecular aggregates in solutions of short-chain substrates, as evidenced by a spectral dye method, likely accounts for the activity of lipases against soluble esters . Unlike esterases, lipases hydrolyze emulsions of water-insoluble medium- and long-chain vinyl esters and TAG such as vinyl laurate, trioctanoin, and olive oil . In conclusion, comparisons of the kinetic behavior of carboxyl ester hydrolases against solutions and emulsions of vinyl esters and TAG allows the distinction between lipases and esterases . In this respect, it clearly appears that guinea pig pancreatic lipase and cutinase are unambiguously classified as lipases. Water Sci Technol, 2002, 46(1-2), 195 - 8 Density separation and molecular methods to characterize enhanced biological phosphorus removal system populations; Schuler AJ et al.; A novel approach to the identification of microorganisms that accumulate high density microbial storage products based on density separation, denaturing gradient gel electrophoresis (DGGE), and DNA sequencing was developed and applied to bench and pilot scale enhanced biological phosphorus removal (EBPR) systems . Polyphosphate (PP), glycogen, and polyhydroxyalkanoates (PHAs), are all of higher density than a typical bacterial cell . PP-accumulating organisms (PAOs), the organisms responsible for EBPR, accumulate all three of these storage products . Density separation in a homogenous solution of Percoll produced a high-density biomass fraction with a relatively high concentration of PAOs, as determined by Neisser staining . DNA was extracted from these fractions, amplified, and separated by DGGE . DGGE profiles demonstrated some bacterial strains were present at a greater concentration in the high density fractions than in low density fractions . These strains were considered PAO candidates . 5 of 12 PAO candidates from high density fractions were gamma Proteobacteria and only 1 was a beta Proteobacterium . 2 PAO candidates were most similar to recently identified gamma Proteobacteria sequences obtained by DGGE analysis of a deteriorated benchtop EBPR system. Waste Manag, 2002, 22(6), 667 - 75 Bioleaching of zinc and aluminium from industrial waste sludges by means of Thiobacillus ferrooxidans; Solisio C et al.; Biological solubilisation of heavy metals contained in two different kinds of industrial wastes was performed in batches employing a strain of Thiobacillus ferroxidans . The wastes tested were: a dust coming from the iron-manganese alloy production in an electric furnace (sludge 1) and a sludge coming from a process treatment plant of aluminium anodic oxidation (sludge 2) . The experimental results pointed out the ability of the used strain to maintain the environment, that initially has a pH about 8, at strongly acid conditions (pH 2.5-3.5), producing sulphuric acid that is the chemical agent responsible for the metals solubilisation . At wastes initial concentration of 1%, the percentage of solubilised metals was 76 and 78% for the wastes 1 and 2, respectively, but the lag phase was considerably longer for sludge 2 than for sludge 1, indicating a different affinity of microorganisms for the solid phase . Increasing the initial slurry concentration, the percentage of removed metal reached 72-73% for the sludge 1, while in case of sludge 2, the total amount of solubilized metal progressively decreased . Two kinetic models are proposed to describe the trends of metals solubilization curves. Int J Vitam Nutr Res, 2002 Jul, 72(4), 251 - 6 Differences in the accumulation of ascorbic acid in normal, myeloperoxidase deficient and NADPH-oxidase deficient granulocytes; Gerber CE et al.; Granulocytes contain large quantities of ascorbic acid (AA) . The uptake mechanism is mainly restricted to the accumulation of the oxidized form, dehydroascorbate (DHA) . We investigated the uptake of ascorbic acid and dehydroascorbate of normal, myeloperoxidase (MPO)-deficient, and NADPH-oxidase-deficient granulocytes . The accumulation of ascorbic acid was increased in all types of granulocytes after stimulation with phorbol-myristate-acetate, whereas the NADPH-oxidase-deficient cells showed a decreased uptake compared to normal and MPO-deficient cells . The intracellular concentration of ascorbic acid was further enhanced after incubation of granulocytes with DHA, most prominently in NADPH-oxidase-deficient granulocytes . MPO-deficient granulocytes are not able to produce HOCl after activation . The granulocytes of one individual with total MPO deficiency accumulated ascorbate in higher concentrations than did cells with partial MPO deficiency, indicating that HOCl is of minor importance for the oxidation of ascorbate . Since the ability of MPO-deficient cells to kill microorganisms is pronounced in contrast to NADPH-oxidase-deficient cells, effective mechanisms of compensating for the absence of HOCl must exist . We hypothesize that the enhanced uptake of ascorbic acid combined with an enhanced superoxide anion production may favor the generation of OH radicals via the Fenton reaction. J Immunol Methods, 2002 Oct 1, 268(1), 107 - 21 CD1 tetramers: a powerful tool for the analysis of glycolipid-reactive T cells; Sidobre S et al.; CD1 proteins constitute a third class of antigen-presenting molecules . They bind lipids rather than peptides, and the T cells reactive to lipids presented by CD1 have been implicated in the protection against autoimmune diseases and infectious microorganisms and in the immune surveillance for tumors . Thus, the ability to identify, purify, and track the response of CD1-reactive cells is of paramount importance . Previously existing methods for identifying these T cells were not based on TCR specificity, and therefore the data obtained by these methods were in some cases difficult to interpret . The recent generation of tetramers of alpha-galactosyl ceramide (alpha-GalCer) with CD1d has already permitted significant insight into the biology of NKT cells . Tetramers constructed from other CD1 molecules also have been obtained during the previous year . Collectively, these new reagents promise to greatly expand knowledge of the functions of lipid-reactive T cells, with potential use in monitoring the response to lipid-based vaccines and other treatments and in the diagnosis of autoimmune diseases. Mol Immunol, 2002 Sep, 39(1-2), 39 - 43 Novel characterisation of the gene encoding conglutinin reveals that previously characterised promoter corresponds to the CL-43 promoter; Hansen S et al.; Conglutinin, a collectin found in bovine serum, is an opsonin that binds to glycoconjugates on the surface of microorganisms or on deposited iC3b, and acts in concert with phagocytes to establish a first-line of immune defense . We have isolated a genomic conglutinin phage clone and found that the 5'-flanking region shows 95.8% identity with the sequence previously published, which on the other hand shows 99.7% identity with the CL-43 promoter . We conclude that the previously published promoter corresponds to the CL-43 promoter and that the functional studies performed on it apply to transcription of CL-43 . Comparison of potential cis-regulatory elements in relation to the functional studies indicates that the two genes are regulated by different mechanisms. Exp Cell Res, 2002 Sep 10, 279(1), 118 - 27 Induction of EGF-dependent apoptosis by vacuolar-type H(+)-ATPase inhibitors in A431 cells overexpressing the EGF receptor; Yoshimoto Y et al.; The stimulation of human tumor cells overexpressing epidermal growth factor receptor (EGFR) with EGF enhances tumor development and malignancy . Therefore, compounds that modulate the EGF-mediated signal inducing apoptosis in EGFR-overexpressing cells would represent a new class of antitumor drug and might be useful in the treatment of a subset of human tumors . In the course of screening for compounds that induce apoptosis in EGFR-overexpressing human epidermal carcinoma A431 cells from secondary metabolites of microorganisms, we found that vacuolar-type H(+)-ATPase (V-ATPase) inhibitors, such as concanamycin B and destruxin E, induced apoptosis only when the cells were stimulated with EGF . The EGF-dependent apoptosis by V-ATPase inhibitors was not observed in other types of human tumor cells which do not overexpress EGFR . The apoptosis in A431 cells was inhibited by anti-FasL antibody which neutralized the cytotoxic effect of FasL, indicating that the Fas/FasL system was involved . The expression of cell surface FasL was upregulated by stimulation with EGF and increased further by V-ATPase inhibitors . Moreover, EGF inhibited cytotoxic Fas antibody-induced apoptosis, whereas V-ATPase inhibitors disrupted the protective effect of EGF on apoptosis in A431 cells . Taken together, these results suggested that V-ATPase inhibitors induced EGF-dependent apoptosis in A431 cells, possibly through both the enhancement of EGF-induced cell surface expression of FasL and the disruption of an EGF-induced survival signal.
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