Pathol Biol (Paris), 1996 Jan, 44(1), 36 - 41 {Modulation of the oxidative burst of human neutrophils by pro- and anti-inflammatory cytokines}; Gougerot-Podicalo MA et al.; Human polymorphonuclear neutrophils (PMN) and cytokines play a critical role in host defences against invading microorganisms . In response to a variety of stimuli, PMN are a major source of reactive oxygen species (ROS) which are essential for bacterial killing and may induce oxidative stress in tissue environment . A precise regulation of the oxidase activity is therefore necessary . Cytokines such as TNF alpha, GM-CSF, IL-8, IL-6, IL-1 alpha and IL-1 beta produced during the immune and inflammatory responses to pathogens have been reported to interact with PMN activities . However, contradictory results have been reported on their direct and priming effects on the PMN release of ROS (oxidative burst) . We have used a flow cytometry method to study the effects of these cytokines on the oxidative burst of PMN in whole blood, in order to avoid PMN activation related to isolation procedures . None of the cytokines tested directly activated the PMN oxidative burst, but they did have differential priming effects on the oxidative burst in response to N-formyl peptides . TNF, GM-CSF and IL-8 strongly primed a subpopulation of PMN to produce H2O2 in response to fMLP, while IL-1 alpha, IL-1 beta and IL-6 failed to do so . Furthermore, the addition of TNF, GM-CSF or IL-8 to whole blood increased the capacity of a subpopulation of PMN to bind N-formyl peptides, a phenomenon that could account for the strong H2O2 production in response to fMLP following priming by the cytokines . These results show that, among the various cytokines tested, TNF, GM-CSF and IL-8 strongly prime the PMN oxidative burst in response to bacterial peptides in whole blood and suggest that these cytokines may play a critical role in bacterial killing in vivo and also in the surrounding tissue injury secondary to pathological inflammatory reactions . In particular, TNF and IL-8 plasma levels as well as LPS-induced monocytic production of these cytokines ex vivo have been correlated with the production of ROS by stimulated PMN and with the lung injury score in patients with Adult Respiratory Distress Syndrom (ARDS) . However, desensitization phenomena have also been described . In particular, in HIV infected patients we demonstrated a decrease of H2O2 production by PMN in whole blood after ex vivo priming by IL-8 and TNF followed by fMLP stimulation . This decrease increased with the progression of the disease and was inversely correlated with IL-8 plasma level . Different mechanisms could explain such desensitization phenomena at the receptor and post receptor level . In addition cytokines are involved in a complex network of regulation and anti inflammatory cytokines, such as IL-10, could act as a negative signal on the proinflammatory cytokines induced-priming of oxidative burst. Biosens Bioelectron, 1996, 11(5), 455 - 77 Microbial detection; Hobson NS et al.; There is a widespread need for commercial instrumentation for the rapid and inexpensive detection of microbial contamination of food, industrial waste water and clinical samples . A large number of detection methods have been developed utilizing the optical, electrochemical, biochemical and physical properties of microorganisms . The need for a device which can produce a rapid, accurate, sensitive, real-time analysis for clinical, industrial and environmental applications has led to considerable progress being achieved in recent years in the development of biosensors for microbial detection . This intense research has resulted in the commercialization of several instruments . Techniques used for the quantification of microorganisms are reviewed under the general categories of non-bioelectrochemical and bioelectrochemical methods. Blood Purif, 1996, 14(1), 102 - 8 Plasma exchange in patients with septic shock including acute renal failure; Stegmayr BG; In patients with septic shock and multiorgan failure including acute renal failure, the prognosis is poor . This study evaluated the effects of plasma exchange as adjunctive therapy to the conventional treatment in such severely ill patients and compared the results with the expected outcome according to data from other studies . A total of 25 patients (17 men and 8 women, mean age 47, range 15-74 years) were treated by a median of 3 (range 1-10) plasma exchanges, mainly by centrifugation technique . The main replacement fluid was liquid-stored plasma and albumin (in a few cases fresh frozen plasma) . About 80% of the patients also received low doses of steroids (hydrocortisone 200-400 mg/day and heparin 1,000-25,000 U/day) . Twenty of the patients survived (80%) which was significantly better than was expected by conventional treatment (< 20% survival, p < 0.001) . The relative chance to survive was four times greater than expected . Five patients died . The reason of death was cerebral hemorrhagia, brain abscess, myocardial sudden death, relapsing sepsis from multiple hepatic abscesses, and a psoas abscess which was not drained . The patients who survived regained almost total renal function and could leave the hospital with only a few sequelae . The plasma exchange may be a good adjunct to conventional therapy to increase the chance of survival when other treatment is insufficient . The mechanisms by which plasma exchange acts may be by removal of endotoxins derived from microorganisms or tissue necrosis and removal of excessive amounts of cytokines together with a modification of various cascade systems enabled by the products in the plasma from the healthy donors, used for replacement. Blood Purif, 1996, 14(1), 94 - 101 Plasmapheresis in severe sepsis or septic shock; Stegmayr BG; During sepsis, there is release of various endotoxins from microorganisms which more or less activates cascade systems including release of cytokines such as tumor necrosis factor alpha and interleukin 6 and complement components . This causes impairment of vascular integrity and permeability which may progress into septic shock and a disseminated intravascular coagulation which progresses into multiorgan failure, including acute renal failure and subsequent death . Although most endotoxins and cytokines have a molecular size < 50 kD, there is little efficacy in removal of them by hemofiltration filters used for acute dialysis . The use of antibodies against different endotoxins has not been successful . The use of plasma exchange procedures (including blood exchange) to remove such toxins and cell debris, as free myoglobin and hemoglobin, has been successfully tried in smaller not controlled studies since 1984 . Once when more than three organs are involved in a progressive manner, the risk of death is at least 80% . In contrast, these studies showed a survival rate of about 75% by addition of such therapeutic interventions to the conventional intensive care unit treatment . The substitution of the removed plasma products must be considered to include products important for the host defense and coagulation process and to avoid infections, bleeding, or increased coagulation . This type of removal is unselective and probably in the future will include addition of absorption techniques which may add further benefit to the outcome. Avian Dis, 1996 Jan-Mar, 40(1), 246 - 51 Mycobacterium genavense infection in canaries; Ramis A et al.; A case of mycobacteriosis in a collection of canaries (Serinus canarius) is described . The affected birds showed nonspecific clinical signs and macroscopic lesions (slight splenomegaly) . Histologic lesions found in liver, spleen, lungs, and kidneys consisted of noncaseous nodules containing accumulations of large macrophages that showed a highly vacuolated cytoplasm in which numerous acid-fast organisms were detected . Attempts to isolate and culture the organisms using Lowenstein-Jensen and Coletsos media proved unsuccessful . Microorganisms belonging to the species Mycobacterium genavense were identified by means of polymerase chain reaction techniques in hepatic tissue from both birds . This finding confirms the existence of fastidiously growing mycobacterial infections, other than Mycobacterium avium, in birds. Acta Vet Scand, 1996, 37(1), 49 - 63 Welfare in Danish dairy herds 1 . Disease management routines in 1983 and 1994; Alban L et al.; This paper presents the first part of a questionnaire survey carried out in 2148 Danish dairy herds during 1994, as well as results from a similar survey carried out in 1983 . The welfare status in Danish dairy herds with respect to disease management routines currently applied is discussed . In detail this was: recording of mastitis incidents, use of veterinarian for milk fever cases, farmer's effort in reducing incidence of mastitis, milk fever, ketosis, calving problems, and lameness, as well as frequency of claw trimming, reasons for culling, and way of replacing cullings . Furthermore, trends during the 11 year period are discussed . The results show that the Danish dairy farmers in 1994 in general have a substantial knowledge of prevention and treatment of disease . However, adjustments in the following areas would be appropriate: 1) farmers should avoid making intravenous infusions, 2) they should be encouraged to use calving boxes for parturitions, 3) there should be more attention on claw health, and 4) to comply with the new Danish legislation, antibiotic dry cow treatment should only be carried out on the individual cow if pathogenic microorganisms have been isolated within 35 days prior to drying off. Int J STD AIDS, 1996 Jan-Feb, 7(1), 61 - 4 Coinfection with chlamydia and gonorrhoea among pregnant women and bacterial vaginosis; Joesoef MR et al.; The role of sexual transmission of microorganisms in bacterial vaginosis (BV) is controversial . If sexual intercourse were a risk factor for BV, then we would expect that women with BV would also be coinfected with other sexually transmitted diseases (STD) . We investigated the prevalence of STD among pregnant women a low socio-economic status with bacterial vaginosis in Indonesia . Among these women, 23.3% had at least one STD (chlamydia, gonorrhoea, syphilis or trichomoniasis) . Chlamydial infection was the most prevalent (19.5%), followed by trichomoniasis (3.8%), gonorrhoea (3.2%) and syphilis (0.4%) . Compared to the rates of STD observed in a previous study of all pregnant women (with or without BV) in Indonesia, pregnant women with BV have more than a 2-fold increase in chlamydia (19.5% vs 8.2%) and a 6-fold increase in gonorrhoea (3.2% vs 0.5%) . Because detection of BV by Gram stain is easy to perform and economical, detection of BV has potential as a prescreening marker for chlamydia and gonorrhoea among asymptomatic pregnant women of low socio-economic status in Indonesia . Further work is needed to evaluate the usefulness of BV as a prescreening marker for chlamydia and gonorrhoea. Ann Pathol, 1996, 16(1), 41 - 4 {Extra-pulmonary pneumocystosis in the course of AIDS . Report of a case}; Terris B et al.; We report a case of splenic pneumocystosis in a human immunodeficiency virus-positive individual treated prophylactically with aerosolized pentamidine . Despite this infection with Pneumocystis carinii, the bronchoalveolar lavage revealed no microorganisms . The use of aerosolized pentamidine as prophylaxis for Pneumocystis carinii is not protective against extrapulmonary pneumocystosis because of inadequate systemic distribution of the drug. Rev Stomatol Chir Maxillofac, 1996, 97(1), 3 - 6 {Primary tuberculosis of the oral cavity}; Junquera Gutierrez LM et al.; Primary tuberculosis in the oral cavity is a rare entity . Usually, the microorganisms need a disruption of the oral mucosa to become pathogenic . In this article the authors describe a clinical case of primary oral tuberculosis, on a female of 52 years-old who suffered an exodontia 20 days before . The bacteria identificated was Mycobacterium tuberculosis hominis . The microbiologic identification is essential to assure the efficacy of the treatment. Adv Biochem Eng Biotechnol, 1996, 54, 75 - 108 Reaction engineering methods to study intracellular metabolite concentrations; Weuster-Botz D et al.; The analysis of intracellular metabolite concentrations is of basic importance for metabolic engineering of microorganisms . In vivo NMR-spectroscopy as a non-invasive technique to measure intracellular metabolite concentrations and rapid sampling devices as invasive techniques are reviewed . The methods are discussed from a reaction engineering point of view . The objective is to obtain intracellular concentration data under well defined physiological conditions in balanced steady state and defined transitional states as well . Application examples are given for a membrane-cyclone-reactor configuration designed to achieve high signal sensitivity with in vivo 31P-NMR and 13C-NMR spectroscopy as well as for a sampling tube device designed for high sampling rates (2s-1) . This sampling device enables the measurement of dynamic metabolite profiles at a time scale of a few seconds. Life Sci, 1996, 58(13), PL249 - 55 Inducible nitric oxide synthase in uterine smooth muscle; Nakaya Y et al.; The expression of inducible nitric oxide synthase (iNOS) mRNA in rat uterus upon in vivo stimulation with lipopolysaccharide (LPS) was studied by reverse transcription and polymerase chain reaction . The injection of LPS induced an increase in mRNA levels of a macrophage-type iNOS . In unstimulated rats, low levels of iNOS mRNA was detected in the uterus and lungs, but absent or negligible in the kidneys and liver . NO was produced in the LPS-treated uterus by addition of 1 to 1000 microM L-arginine . The production of NO in uterine tissue that faces the outside of the body may provide a bacteriocidal protective function against microorganisms in physiological condition . However, NO produced in a large amounts by cytokine and LPS may play some pathological reaction during septic shock or infection. FEMS Microbiol Lett, 1996 Jan 1, 135(1), 37 - 43 Identification of a Renibacterium salmoninarum DNA fragment associated with bacterial internalization into CHSE-cultured cells; Maulen NP et al.; We report here the isolation of a Renibacterium salmoninarum DNA sequence capable of transforming a non-invasive Escherichia coli strain into a microorganism able to enter the fish cell line, CHSE-214 . Immunofluorescence and electron microscopy techniques were used to assess the acquired invasive phenotype by HB101 E . coli cells, upon transformation with pPMV-189 . This plasmid carries a 2282-bp R . salmoninarum DNA segment . The invasive phenotype is conserved upon deletion of approximately 1000 bp at the 3' end of the insert . The remaining segment contains an ORF region encoding a putative protein of about 30 kDa. Toxicol Lett, 1996 Jan, 84(1), 33 - 6 Characterization of 2,4-dichlorophenoxyacetic acid transport and its relationship with polyamines in Azospirillum brasilense; Castro S et al.; We have previously shown that 2,4-dichlorophenoxyacetic acid (2,4-D) inhibits Azospirillum brasilense growth, the synthesis of DNA, RNA and proteins . These toxic effects are prevented when polyamines are added to the culture medium . The purposes of our research were to determine the effects of the herbicide on the number of viable Azospirillum brasilense cells, characterize the 2,4-D transport system and to study the effects of polyamines upon the latter in this microorganism . We found that 2,4-D reduced the number of viable cells and that 2,4-D transport is energy-independent, since it was not affected by metabolic inhibitors . Polyamines did not alter 2,4-D uptake, further supporting the hypothesis that the herbicide most likely produces its toxic effects by interfering with the polyamine metabolism. Can J Microbiol, 1996 Jan, 42(1), 79 - 82 Isolation and description of carbazole-degrading bacteria; Shotbolt-Brown J et al.; Carbazole is a nitrogen heterocyclic compound associated with fossil fuels and their products . Enrichment cultures were established to isolate bacteria able to degrade carbazole and a plate assay was developed to select carbazole-degrading microorganisms from the enrichments . Three different bacterial isolates capable of mineralizing carbazole were obtained . No intermediates of carbazole degradation were detected . The bacteria had a limited substrate specificity; all used benzoate for growth but were unable to utilize the analogues of carbazole, fluorene, or dibenzothiophene. Trends Biotechnol, 1996 Jan, 14(1), 6 - 12 Isolating and characterizing deep-sea marine microorganisms; Kato C et al.; We have isolated several microorganisms that are adapted to living in the extremes of the deep-sea environment . They include barophilic bacteria, which are able to grow at high hydrostatic pressure, but that are unable to grow at atmospheric pressure, and organic-solvent-tolerant bacteria, which are able to grow in the presence of toxic organic solvents such as toluene or benzene . In this review, we describe how to isolate such extremophiles, and we outline the characteristics of several strains that have been recovered from the deep-sea environment. Arch Environ Contam Toxicol, 1996 Jan, 30(1), 107 - 13 Liver microsomal levels of cytochrome P450IA1 as biomarker for exposure and bioavailability of soil-bound polycyclic aromatic hydrocarbons; Roos PH et al.; The bioavailability of soil-bound polycyclic aromatic hydrocarbons (PAHs) for mammalian species was studied with rats fed with a diet containing contaminated soil preparations . The extent of cytochrome P450IA1 (CYP1A1) induction in the liver correlated with the amount of 5- and 6-ring PAHs in the soil samples but not with the total PAH content . Other cytochromes P450 were much less affected by the soil-contaminants . The highest induction of CYP1A1 was obtained with a sample containing 274 mg 5- and 6-ring PAH/kg soil, resulting in a nearly 360-fold increase in the ethoxyresorufin deethylase (EROD) activity . In a semilogarithmic plot, a linear correlation was found between the 5- and 6-ring PAH concentration in the soil and the microsomal CYP1A1 content . As a model for the action of intestinal fluids, soil samples were extracted by bile acid solution . In these experiments, the selectivity in the solubilization of individual PAHs parallels that of toluene extraction, although the yield is lower than the latter and varies with the soil sample . The bioavailability of PAHs for microorganisms, but not for mammals, was shown to be considerably reduced in the presence of high total organic carbon (TOC) values of the soil samples . This may have implications for decontamination strategies, diminishing the effectiveness of biological decontamination in cases with high TOC values . The data suggest that CYP1A1 induction in rats is a parameter that may be useful in risk assessments of contaminated soils for mammalian species. Int J Syst Bacteriol, 1996 Jan, 46(1), 167 - 72 Analysis of the genetic polymorphism of Borrelia burgdorferi sensu lato by multilocus enzyme electrophoresis; Balmelli T et al.; In recent years, Borrelia burgdorferi sensu lato has been subdivided into three species, Borrelia burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii, and a new species restricted to Japan, Borrelia japonica, has been isolated from Ixodes ovatus . In addition, members of several new genomic groups have been found in America and in Europe, suggesting that there are additional genospecies . In order to study the diversity of B . burgdorferi sensu lato, we analyzed 54 isolates, cultured from humans and from different tick species and obtained from diverse geographic areas, including Europe, the United States, Japan, and the People's Republic of China . In order to investigate the genetic relationship between microorganisms that are transmitted by soft ticks and microorganisms that cause Lyme disease, we also included three strains of relapsing fever spirochetes . The method which we used was multilocus enzyme electrophoresis; 12 genetic loci were characterized on the basis of the electrophoretic mobilities of their products, and 50 distinct allele profiles (electrophoretic types) were distinguished . The mean genetic diversity per locus was 0.747 . a cluster analysis of a matrix of genetic distances for pairs of electrophoretic types revealed 11 divisions that were separated at genetic distances greater than 0.65 . Five of these divisions corresponded to B . burgdorferi sensu stricto, B . garinii, B . afzelii, B . japonica, and the newly proposed species inverted question markBorrelia andersonii . inverted question mark Our results also confirmed that there are two additional genomic groups in Europe and at least one additional group in the United States . The relapsing fever spirochetes were were not clearly separated from the spirochetes associated with Lyme disease . In conclusion, we believe that the previously proposed subdivision of B . burgdorferi sensu lato into only four species should be reconsidered. FASEB J, 1996 Jan, 10(1), 49 - 56 In vitro folding of inclusion body proteins; Rudolph R et al.; Insoluble, inactive inclusion bodies are frequently formed upon recombinant protein production in transformed microorganisms . These inclusion bodies, which contain the recombinant protein in an highly enriched form, can be isolated by solid/liquid separation . After solubilization, native proteins can be generated from the inactive material by using in vitro folding techniques . New folding procedures have been developed for efficient in vitro reconstitution of complex hydrophobic, multidomain, oligomeric, or highly disulfide-bonded proteins . These protocols take into account process parameters such as protein concentration, catalysis of disulfide bond formation, temperature, pH, and ionic strength, as well as specific solvent ingredients that reduce unproductive side reactions . Modification of the protein sequence has been exploited to improve in vitro folding. J Bacteriol, 1996 Jan, 178(2), 385 - 95 Induction of synthesis of tetrahydropyrimidine derivatives in Streptomyces strains and their effect on Escherichia coli in response to osmotic and heat stress; Malin G et al.; The metabolic responses of a number of Streptomyces strains to osmotic and heat stress were studied by 13C nuclear magnetic resonance spectroscopy . During cell growth in a chemically defined medium supplemented with 0.5 M NaCl, tetrahydropyrimidine derivatives (THPs), 2-methyl-4-carboxy-5-hydroxy-3,4,5,6-tetrahydropyrimidine {THP(A)} and, to a lesser extent, 2-methyl-4-carboxy-3,4,5,6-tetrahydropyrimidine {THP(B)}, were found to accumulate in a significant amount in all bacteria examined . In addition, when the growth temperature was shifted from 30 to 39 degrees C, the intracellular concentration of THP(A) increased significantly . Moreover, exogenously provided THP(A) or THP(B) or both reversed inhibition of Escherichia coli growth caused by osmotic stress and increased temperature . Although the ability of Streptomyces strains to tolerate high concentrations of NaCl is well known, very little is known about the osmoregulatory strategy in Streptomyces strains . Similarly, the mechanism by which compatible solutes accumulate in a variety of microorganisms is not understood . Our findings suggest the possibility of a novel mechanism of protection of DNA against salt and heat stresses involving the THPs. Am J Pathol, 1996 Jan, 148(1), 95 - 104 Immunomagnetic separation can enrich fixed solid tumors for epithelial cells; Yaremko ML et al.; Immunomagnetic separation is a highly specific technique for the enrichment or isolation of cells from a variety of fresh tissues and microorganisms or molecules from suspensions . Because new techniques for molecular analysis of solid tumors are now applicable to fixed tissue but sometimes require or benefit from enrichment for tumor cells, we tested the efficacy of immunomagnetic separation for enriching fixed solid tumors for malignant epithelial cells . We applied it to two different tumors and fixation methods to separate neoplastic from non-neoplastic cells in primary colorectal cancers and metastatic breast cancers, and were able to enrich to a high degree of purity . Immunomagnetic separation was effective in unembedded fixed tissue as well as fixed paraffin-embedded tissue . The magnetically separated cells were amenable to fluorescence in situ hybridization and polymerase chain reaction amplification of their DNA with minimal additional manipulation . The high degree of enrichment achieved before amplification contributed to interpretation of loss of heterozygosity in metastatic breast cancers, and simplified fluorescence in situ hybridization analysis because only neoplastic cells were hybridized and counted . Immunomagnetic separation is effective for the enrichment of fixed solid tumors, can be performed with widely available commercial antibodies, and requires little specialized instrumentation . It can contribute to interpretation of results in situations where enrichment by other methods is difficult or not possible. Biochem Biophys Res Commun, 1995 Dec 26, 217(3), 1255 - 62 PI 3-kinase-dependent and independent chemotaxis of human neutrophil leukocytes; Thelen M et al.; The migration of neutrophil leukocytes to inflammatory sites is important for the elimination of microorganisms but can under pathological conditions lead to severe tissue damage . The initial chemotactic response is elicited by classical chemoattractants, such as fMet-Leu-Phe or the chemokine interleukin-8 which ligate to G-protein coupled receptors . Neutrophils show also a delayed chemotactic response to growth factors, such as platelet derived growth factor (PDGF) or tumor growth factor (TGF beta) . We describe here that classical chemoattractants and growth factors stimulate neutrophil chemotaxis through different signal transduction pathways . Wortmannin, a selective phosphatidylinositol 3-kinase inhibitor, completely blocks growth factor stimulated chemotaxis while having no effect on neutrophil migration stimulated with classical chemoattractants . The results suggest that cell migration can be selectively controlled through the inhibition of distinct signal transduction events. Arch Biochem Biophys, 1995 Dec 20, 324(2), 273 - 81 Characterization of the antifungal protein secreted by the mould Aspergillus giganteus; Lacadena J et al.; An antifungal polypeptide (AFP) of 51 amino acid residues, secreted by the mould Aspergillus giganteus, has been purified to homogeneity and characterized . The inhibitory effect of this protein on the growth of different microorganisms has been studied . Whereas the growth of many of the filamentous fungi assayed is inhibited, no effect has been observed against yeasts or bacteria . The minimal concentration for total inhibition of the growth is in the range 6 to 25 microM . The antifungal polypeptide does not produce any effect on the growth of the producing mould . The polypeptide promotes aggregation of acidic phospholipid vesicles . A remarkable resistance to proteolysis and a low hydrogen x deuterium exchange have been observed for this protein . The protein does not show any thermal transition up to 80 degrees C when studied by differential scanning calorimetry and infrared spectroscopy . The uv absorbance, fluorescence emission, and circular dichroism (CD) characteristics of this protein have been studied . The protein exhibits a strong positive band at 230 nm as a prominent feature of the CD spectrum in the far uv region . All the spectroscopical properties of the antifungal protein are highly influenced by the abundance of tyrosine residues . These can be grouped in two different populations, buried and exposed, based on the results of pH-titration experiments . Fourier-transform infrared spectroscopy reveals a high content of beta-structure in AFP . Reduction and carboxy-amidomethylation produces a rather unstructured polypeptide as deduced from its spectroscopical properties. Biochim Biophys Acta, 1995 Dec 20, 1241(3), 407 - 23 The epithelial mucin, MUC1, of milk, mammary gland and other tissues; Patton S et al.; MUC1 is a mucin-type glycoprotein that is integrally disposed in the apical plasma membrane of the lactating epithelial cell and protrudes from the cell surface into the alveolar lumen where milk is stored . Envelopment of milk fat globules by this membrane accomplishes their secretion and conveys MUC1 into milk . The human form of this mucin has been detected in many other organs, tissues and body fluids . It projects from the cell surface as long filaments . In the human and a number of other species, MUC1 is polymorphic due to variable numbers of a tandemly repeated segment 20 amino acids in length . The individual codominantly expresses two alleles for the mucin so that differences in its size among individuals and between the two forms of an individual are observed . The tandem repeats are rich in serines and threonines which serve as O-glycosylation sites . Carbohydrate content of MUC1, as isolated from milk of human, bovine and guinea pig, is approximately 50% . The oligosaccharides carry substantial sialic acid at their termini and this accounts for two putative functions of this mucin, i.e., to keep ducts and lumens open by creating a strong negative charge on the surface of epithelial cells which would repel opposite sides of a vessel, and to bind certain pathogenic microorganisms . MUC1 is protease resistant (trypsin, chymotrypsin and pepsin) and large fragments of it can be found in the feces of some but not all breast-fed infants . MUC1 has a highly varied structure because of its polymorphism, qualitative and quantitative variations in its glycosylation between tissues, individuals and species, and differences due to divergence in the nucleotide sequences among species . Sequencing of the MUC1 gene for various species is showing promise of revealing unique evolutionary relationships and has already indicated conserved aspects of the molecule that may be functionally important . Among these are positions of serine, threonine and proline in the tandem repeats and a high degree of homology in the transmembrane and cytoplasmic segments of the molecule. J Immunol, 1995 Dec 15, 155(12), 5671 - 7 Mouse surfactant protein-D . cDNA cloning, characterization, and gene localization to chromosome 14; Motwani M et al.; Surfactant protein-D (SP-D) is a collectin found associated with surfactant in the lung . SP-D has also been functionally characterized as an opsonin for diverse microorganisms and a chemoattractant for phagocytic cells . To determine the structure of mouse SP-D, we isolated and characterized clones from a B6/CBAF1J strain lung cDNA library using a PCR-derived genomic probe . The deduced sequence predicts a 19-amino acid signal sequence, a 25-amino acid long NH2 terminus with two cysteines, followed by an uninterrupted collagen domain with 59 Gly-X-Y repeats . Next, a short "neck" domain of 28 amino acids, with a potential to form trimeric alpha-helical coiled coil is found ending in a COOH-terminal 125-amino acid carbohydrate recognition domain . The mature mouse SP-D protein of 355 amino acids shows strong homology to rat (92% identity), human (76%), and bovine (72%) SP-D amino acid sequences . Northern blot and RT-PCR analysis revealed that the mouse SP-D gene is expressed predominantly in lung and, surprisingly, also in heart, stomach, and kidney but not in brain . In contrast, mouse surfactant protein-A (SP-A) mRNA expression was found to be restricted to lung . Human lung and stomach, but not heart or liver, were found to express SP-D mRNA, as determined by PCR . The mouse SP-D gene (Sftp4) has been localized to chromosome 14 (to a region syntenic to human chromosome 10), closely linked to the genes for other collagenous lectins, mannose-binding protein-A (MbI1), and SP-A (Sftp1). J Biol Chem, 1995 Dec 8, 270(49), 29229 - 35 From xenobiotic to antibiotic, formation of protoanemonin from 4-chlorocatechol by enzymes of the 3-oxoadipate pathway; Blasco R et al.; Chloroaromatics, a major class of industrial pollutants, may be oxidatively metabolized to chlorocatechols by soil and water microorganisms that have evolved catabolic activities toward these xenobiotics . We show here that 4-chlorocatechol can be further transformed by enzymes of the ubiquitous 3-oxoadipate pathway . However, whereas chloromuconate cycloisomerases catalyze the dechlorination of 3-chloro-cis,cis-muconate to form cis-dienelactone, muconate cycloisomerases catalyze a novel reaction, i.e . the dechlorination and concomitant decarboxylation to form 4-methylenebut-2-en-4-olide (protoanemonin), an ordinarily plant-derived antibiotic that is toxic to microorganisms. Ecotoxicol Environ Saf, 1995 Dec, 32(3), 244 - 53 The ecotoxicity of chlorate to aquatic organisms: a critical review; van Wijk DJ et al.; In order to assess the risk posed by chlorate in aquatic ecosystems, data on the effects of chlorate on aquatic organisms (microorganisms, algae, invertebrates, and fish) and mesocosm studies have been collated and critically reviewed . The geometric mean E(L)C50 values for both freshwater and marine species were (as ClO3-): microorganisms, 38,583 mg.liter-1; microalgae, 563 mg.liter-1; invertebrates, 2442 mg.liter-1; fish, 3815 mg.liter-1 . Marine macro red algae were insensitive to chlorate, whereas marine macro brown algae (e.g., Fucus sp.) appeared to be exceptionally sensitive to chlorate, adverse long-term effects having been reported at concentrations as low as 0.015 mg ClO3-.liter-1 . Evidence for the mechanism by which chlorate is thought to be particularly toxic to these species is also reviewed . It is concluded that, based on the species reported, chlorate is nontoxic (acute toxicity > 100 mg.liter-1) to most of the freshwater and marine species examined . However, chlorate is highly toxic (acute toxicity < 0.1 mg.liter-1) to certain macro brown algal species . For macro brown algae, the NOEC after 6 months was reported to be approximately 0.005 mg ClO3-.liter-1 . It is also concluded that an improved understanding of the actual mode of action of chlorate in sensitive species is desirable . Together with further information on the environmental fate of chlorate, this will improve the risk assessment for chlorate in the aquatic environment. Ecotoxicol Environ Saf, 1995 Dec, 32(3), 205 - 8 Effective concentrations of nitrophenolics toward growth yield of selected microalgae and cyanobacteria isolated from soil; Madhavi DR et al.; The effective concentrations (EC), which cause a certain percentage of inhibition in growth yield, of four nitrophenolics (o-nitrophenol, m-nitrophenol, p-nitrophenol (PNP), and 2,4-dinitrophenol) and the major metabolites of PNP reduction (p-nitrosophenol and p-aminophenol) toward growth of two microalgae (Chlorella vulgaris and Scenedesmus bijugatus) and two cyanobacteria (Nostoc muscorum and Nostoc linckia), all isolated from soil, were determined following linear regression analysis . The EC50 values of the selected toxicants toward the test organisms were in the range of 32 to 227 micrograms ml-1 . Such a wide range in EC figures emphasizes the need for a prior determination of EC values of a toxicant before establishing its toxicity pattern toward a nontarget microorganism. Arch Histol Cytol, 1995 Dec, 58(5), 517 - 22 A histochemical and immunohistochemical study of certain defense mechanisms in the human lacrimal sac epithelium; Perra MT et al.; The mucosal surface of the human lacrimal sac represents an area exposed to exogenous agents including potentially harmful microorganisms . The human lacrimal sac was examined histochemically to identify glycoproteins, and immunohistochemically to identify secretory IgA . Neutral and acid glycoconjugates were detected mainly in the cytoplasm of the surface cells of the columnar stratified epithelial lining . The same reactions were recognized in occasional clusters of secretory cells forming intraepithelial glands in the lining of the lacrimal sac . The presence of secretory IgA in the cytoplasm of the apical epithelial cells was demonstrated . The results indicate that the lacrimal sac mucosa possesses certain active defense mechanisms against ascending infections. Electrophoresis, 1995 Dec, 16(12), 2249 - 57 Gaining insight into a complex organelle, the phagosome, using two-dimensional gel electrophoresis; Burkhardt J et al.; Phagosomes are the organelles formed de novo in a variety of cells by the internalization of large particulate materials, including a wide range of pathogenic microorganisms . We present here a systematic approach that can be used to study the polypeptide composition of phagosomes/phagolysosomes and to yield analytical information on the characteristics of their proteins . A density shift approach was used to isolate pure preparations of phagosomes filled with low density latex beads from mouse J774 and human U937 macrophages . High resolution two-dimensional (2-D) gel electrophoresis was performed to generate a map of the overall {35S}methionine-labeled protein profile of the isolated phagosomes . The resulting map showed the minimal presence of over 200 polypeptides, indicating the complexity of this organelle . Comigration experiments showed that several phagosome polypeptides, among them several known proteins, are shared by the two species . Extraction with Triton X-114 and sodium carbonate was performed to distinguish between membrane and soluble proteins, and sensitivity to a panel of proteases was measured to identify proteins exposed on the cytoplasmic face of the phagosome membrane . The general value of the 2-D gel approach in the mapping of organelle proteins is discussed. Lymphology, 1995 Dec, 28(4), 196 - 207 Immune surveillance of the oral cavity and lymphocyte migration: relevance for alcohol abusers; Chiappelli F et al.; The health of the oral cavity is threatened by a variety of microorganisms . Impaired immune surveillance of the oral environment contributes to the development of infectious processes and tumors of the mouth . Elucidation of the physiological mechanisms that determine and control oral immune surveillance is crucial to an understanding of these oral diseases . The ability of lymphocytes to migrate is critical for successful immune surveillance . the cardinal facets of lymphocyte migration are reviewed here in the context of the oral cavity . One mechanism by which alcohol acts as an important cofactor in the onset and development of oral diseases is hypothesized to be through impaired lymphocyte migration to and from peri-oral lymphoid tissue. Cytopathology, 1995 Dec, 6(6), 388 - 402 Morphological study of bacteria of the respiratory system using fluorescence microscopy of Papanicolaou-stained smears with special regard to the identification of Mycobacteria sp; Kupper T et al.; In Papanicolaou-stained smears certain structures such as nucleoli, Pneumocystis carinii, Charcot-Leyden crystals, bacteria and fungi show a brilliant fluorescence . The morphological characteristics of microorganisms which can be detected by this system, especially mycobacteria, are described . This screening method offers the possibility of providing the clinician with a provisional diagnosis within hours . Proof of the nature of the organisms should be obtained by culture. Semin Cell Biol, 1995 Dec, 6(6), 357 - 65 NADPH oxidase and the respiratory burst; Wientjes FB et al.; Phagocytic cells possess an electron-transport system which accepts electrons from NADPH in the cytosol to reduce oxygen to the superoxide radical in the vacuolar lumen . The superoxide is instrumental in killing ingested microorganisms . Patients suffering from chronic granulomatous disease (CGD), in which this system is failing, are abnormally susceptible to infectious diseases . Studying CGD patients' neutrophils has been enormously helpful in identifying the components of the superoxide-generating system, known as the NADPH oxidase . This review will describe the components of the electron-transport chain involved in the oxidase and the factors needed for its regulation. Minerva Ginecol, 1995 Dec, 47(12), 557 - 60 {Genital infections as a cause of abortion in the first trimester of pregnancy . Review of the literature}; Capuzzo E et al.; The rate of spontaneous abortion from embryo-fetal infection is believed to range from 10 to 15% . This figure, however, is likely to be underestimated because of subclinical abortions occurring at the earliest stages of gestation that go undiagnosed . The actual frequency of infections resulting in 1st trimester spontaneous abortion is therefore unknown, whereas the impact of infectious agents in the TORCH group on the pathogenesis of such event is well known, although the pathogenetic mechanisms are not well understood . A wide range of microorganisms have been isolated by several investigators from the lower genital tract of women with 1st trimester spontaneous abortion . No causative relationship of HIV-1 infection to spontaneous abortion has been established yet . The diagnosis of abortion from infection can only be made retrospectively based on histologic examination of fetal and placental tissue and isolation on culture of the infectious agent assumed to have caused infection. Mol Biotechnol, 1995 Dec, 4(3), 275 - 95 Two-dimensional DNA typing: A cost-effective way of analyzing complex mixtures of DNA fragments for sequence variations; Vijg J; By two-dimensional (2D) DNA typing, multiple DNA fragments can be analyzed in parallel for all possible sequence variations . The method involves electrophoretic separation in polyacrylamide gels, first by size and subsequently on the basis of basepair sequence in a denaturing gradient . The system has been automated partly and a dedicated image analysis system for rapid interpretation of the often complex spot patterns was developed . In this review, the basic principles of 2D DNA typing, its methodology, and some major applications in genetic studies of animals, plants, and microorganisms will be discussed, with the emphasis on human genetics. Mol Biotechnol, 1995 Dec, 4(3), 227 - 37 Development of a novel, rapid processing protocol for polymerase chain reaction-based detection of bacterial infections in synovial fluids; Mariani BD et al.; We describe the development of a molecular detection system designed for use with synovial fluid (SF)-based infections . The methodology employs a lysis/extraction procedure that effectively disrupts microorganisms allowing for release of the microbial DNA and its amplification by polymerase chain reaction (PCR) . We tested the effectiveness of adding a mixed-bed, ion-exchange resin to the extract to remove PCR inhibitory components present in the SF . After centrifugation to separate the resin, DNA contained in the supernatant is subjected to PCR using oligonucleotide primers designed for broad-spectrum microorganism detection . Amplification products are analyzed by agarose gel electrophoresis and/or DNA hybridization methodology . We report here the detection sensitivity and specificity of the protocol using SF inoculated with Escherichia coli and Staphyloccocus aureus . We have applied this new methodology to clinical SF specimens with results superior to standard laboratory culturing assays. Int Dent J, 1995 Dec, 45(6), 352 - 7 Environmental microbial contamination . Pilot study in a dental surgery; Osorio R et al.; Environmental contamination by bacterial aerosols occurs every day in the dental surgery . The aim of this study was to determine bacterial levels in five different areas of a dental surgery during ultrasonic scaling procedures using bacterial cultures . Two areas with markedly different amounts of infective aerosols were identified . The role of the air conditioning system was also assessed . There was evidence that the air conditioning system could act as a vehicle for the transmission of microorganisms. Rev Sci Tech, 1995 Dec, 14(4), 937 - 50 Risk analysis in the manufacture of veterinary biologicals; Moreau Y; "Primum non nocere' must be the first quality of veterinary immunological medicinal products . Throughout the manufacturing process, stage by stage, various ingredients and/or operations could be responsible for safety problems observed when the product is administered . During the preliminary stages, various ingredients--e.g . master seed strains (virus, bacteria or parasite), cell substrates (cell-lines or primary cells) and substances of animal origin--could be contaminated by extraneous agents . These represent the most important risks . During the process--i.e . preliminary stages, microorganism growth, downstream processing (harvest, purification, concentration, inactivation, etc.), formulation, filling, freeze-drying and finally packaging--environmental conditions (working areas, equipment, etc.) might also be defective and responsible for product contamination . The author examines all aspects of risks and their assessments through consideration of "good manufacturing practice' based on quality assurance and quality control systems. Clin Oral Implants Res, 1995 Dec, 6(4), 197 - 204 The effect of periodontal parameters on the subgingival microbiota around implants; Papaioannou W et al.; The study aimed to examine the relationship between the subgingival flora around implants and their periodontal parameters . Plaque samples from 561 implants (279 patients) were analyzed by means of differential phase contrast microscopy and compared with the sample site's probing depth, bleeding tendency on probing, and plaque and gingivitis indices . If possible, one implant with deep and one with shallow pockets were selected within the same patient . The impact of the intraoral exposure time on the microbial composition around the implants was examined cross-sectionally, with the same group of patients . Only tendencies can be detected by the latter, and no concrete conclusions can be drawn . From the clinical parameters, increased probing depth was found to detrimentally increase the proportion of spirochetes and motile organisms, whereas the other parameters were found to be of minor importance . For partially edentulous patients only, there was a tendency for increased proportions of spirochetes and motile organisms the longer the intraoral exposure time . These observations emphasize the importance of the periodontal health of the remaining teeth (as a reservoir of pathogenic microorganisms) in partial edentulous patients rehabilitated by means of implants and indicate the importance of shallow pockets around implants (flap trimming when aesthetics and phonetics allow). Korean J Parasitol, 1995 Dec, 33(4), 387 - 90 Axenization of Entamoeba histolytica, a Korean strain YS-27; Chang JK et al.; In the present communication, YS-27, a Korean strain of Entamoeba histolytica is described for the isolation and establishment of axenic cultivation . E . histolytica, designated as strain "YS-27", was isolated from the pus of a hepatic abscess obtained from a 72 year old inpatient of August 10, 1969 . Specimens, were obtained by needle aspiration, inoculated immediately and weekly cultured in a modified diphasic medium at 37 degrees C . Strain YS-27 had been maintained for more than 15 years by weekly subculture until February, 1985 . These cultures were transferred to a monoxenic TTY-SB medium seeded with a trypanosomatid of the genus Crithidia . Penicillin G, 2 to 10 x 10(3) International units and Streptomycin, 2 to 10 mg per 100 ml, were added to the cultures to eliminate the bacteria . After more than one year later, these two organisms were well maintained by transfer every 3 or 4 days until January, 1986 at 37 degrees C in TTY-SB medium in the absence of other microorganisms . These monoxenic cultures were then transferred to TYI-S-33 medium . Strain YS-27 alone had not been growing at the time of transfer, but when overlaid with Crithidia at intervals of 3 to 4 days, strain YS-27 propagated well . The Crithidia died out several weeks later after several passages . Beginning in April, 1986, strain YS-27, was successfully established in axenic culture in TYI-S-33 medium and has been maintained in continuous culture and multiplied well to present. Comp Biochem Physiol B Biochem Mol Biol, 1995 Dec, 112(4), 637 - 42 Steroid hormone signalling system and fungi; Zakelj-Mavric M et al.; Three components of the steroid hormone signalling system, 17 beta-hydroxysteroid dehydrogenase, androgen binding proteins and steroid hormone signalling molecule testosterone were determined in the filamentous fungus Cochliobolus lunatus for the first time in a fungus . Their possible role in C . lunatus is discussed in comparison with their role in mammalian steroid hormone signalling system . The results are in accordance with the hypothesis, that the elements of primordial signal transduction system should exist in present day eukaryotic microorganisms. Microbiologia, 1995 Dec, 11(4), 429 - 38 The social behavior of myxobacteria; Munoz-Dorado J et al.; Myxobacteria are social microorganisms that undergo a spectacular cell cycle . Under starvation conditions, cells aggregate to certain points originating macroscopic fruiting bodies, inside which cells differentiate into myxospores . To accomplish this developmental cycle, cells must communicate . The signals that cells exchange during development as well as the signal transduction systems used by myxobacteria have been intensively studied during the last years . A family of eukaryotic-like protein serine/threonine kinases has been identified in Myxococcus xanthus, indicating that signal transduction systems similar to those used by eukaryotic cells may also function in myxobacteria. Cell Tissue Res, 1995 Dec, 282(3), 455 - 61 Selective binding and transcytosis of Ulex europaeus 1 lectin by mouse Peyer's patch M-cells in vivo; Clark MA et al.; The in vivo interaction of the lectin Ulex europaeus agglutinin 1 with mouse Peyer's patch follicle-associated epithelial cells was studied in the mouse Peyer's patch gut loop model by immunofluorescence and electron microscopy . The lectin targets to mouse Peyer's patch M-cells and is rapidly endocytosed and transcytosed . These processes are accompanied by morphological changes in the M-cell microvilli and by redistribution of polymerised actin . The demonstration of selective binding and uptake of a lectin by intestinal M-cells in vivo suggests that M-cell-specific surface glycoconjugates might act as receptors for the selective adhesion/uptake of microorganisms. Pneumologie, 1995 Dec, 49 Suppl 3, 636 - 42 {Mycobacteria and mycobacterioses}; Bottger EC; The genus Mycobacterium harbours a number of significant pathogens . The diagnosis of mycobacterial infections has traditionally relied on microscopical and cultural techniques which were compromised by the slow growth of these microorganisms . More recently, molecular methods suitable for use in diagnostic microbiology have been developed and have been demonstrated to significantly improve both our diagnostic capabilities as well as our understanding of this complex genus of microorganisms. Leber Magen Darm, 1995 Dec, 25(6), 251 - 7 {Preventing transmission of infection in endoscopy: hygienic maintainance of flexible endoscopes and measures for personal protection}; Leiss O et al.; For prevention of transfer of infective agents by the contaminated endoscope the importance of thorough mechanical cleaning of the endoscope and adequate disinfection is stressed . Mode of action and side effects of liquid disinfectants as well as resistance of microorganisms (spores and some mycobacteria) to germicidal chemicals are mentioned . The different steps of disinfection procedures and potential causes of failure are discussed . Automatic disinfection systems are required for a higher degree of security of both patient and staff of the endoscopic unit . A regular control of the efficacy of the disinfection process is recommended and models for implementation are discussed . For prevention of occupationally acquired infection general precaution guidelines (use of gloves, prevention of needle-stick injuries) and vaccination programs are stressed. Am J Physiol, 1995 Dec, 269(6 Pt 1), L721 - 6 Differential regulation of the mannose and SP-A receptors on macrophages; Chroneos Z et al.; Two carbohydrate-dependent mechanisms exist on alveolar macrophages to clear mannose-containing pathogens: receptor-mediated entry of non-opsonized microorganisms via the mannose receptor and receptor recognition of pathogens opsonized with surfactant-associated protein A (SP-A) . A number of studies have demonstrated that mannose receptor expression is tightly linked to the functional state of the macrophage . In the present study, we investigated regulation of binding of SP-A to its receptor on macrophages by the same agents that regulate mannose-receptor expression . Phorbol 12-myristate 13-acetate, lipopolysaccharide (LPS), and interferon-gamma treatment of rat marrow-derived macrophages increased SP-A binding by 163, 296, and 337%, respectively, over untreated controls . Mannose-receptor activity was reduced to 75, 60, and 25% of control levels by these agents . Dexamethasone increased mannose receptor activity to 225%, while decreasing SP-A binding to 44% of controls . Addition of granulocyte macrophage-colony stimulating factor (GM-CSF) to human monocytes on day 0 dramatically increased mannose-receptor activity on day 5 over the non-serum control . SP-A binding was highest to freshly isolated monocytes and decreased to < 10% after differentiation in the presence of GM-CSF . After intraperitoneal injection of dexamethasone, rat alveolar macrophages isolated at 24 h expressed increased mannose-receptor activity and decreased SP-A binding . LPS injection resulted in increased SP-A binding and decreased mannose-receptor activity . In every instance, SP-A binding was inversely regulated with respect to mannose-receptor expression . We therefore speculate that the mannose receptor is a first-line host-defense receptor that is turned off during inflammation . SP-A in the alveolar space can then act as a lung-specific opsonin and mediate clearance of pathogens via the upregulated SP-A receptor. J Biotechnol, 1995 Dec 1, 43(2), 145 - 58 Measurements of sulfur, phosphorus and other ions in microbial biomass: influence on correct determination of elemental composition and degree of reduction; Duboc P et al.; The elemental composition of microorganisms varies widely with respect to the N, S, P and ash content as well as many trace elements depending on microorganism and growth conditions . As a consequence, the mass of 1 C-mol of biomass can differ considerably from one microorganism to another . Experimental results show that sulfur is completely volatilized during combustion, so that the calculation of the mass fraction of oxygen is affected when S is not measured . However, its neglect does not seriously affect the elemental composition and the degree of reduction of biomass . Phosphorus is found in ash after combustion at the same degree of reduction as in biomass . Therefore, oxygen bound to it should be included in the elemental formula, so that the degree of reduction of biomass remains unchanged by inclusion of P into elemental formula . No external oxygen is incorporated into biomass during combustion and thus the measured ash fraction is correct. Can J Microbiol, 1995 Dec, 41(12), 1117 - 23 Alterations in cell pigmentation, protein expression, and photosynthetic capacity of the cyanobacterium Oscillatoria tenuis grown under low iron conditions; Trick CG et al.; To better describe the iron-limited nutrient status of aquatic photosynthetic microorganisms, we examined the effects of iron limitation on pigment content, maximum rates of photosynthetic oxygen evolution, and respiratory oxygen consumption in the filamentous cyanobacterium Oscillatoria tenuis Ag . Within the range of iron (4.2 x 10(-5)-5.1 x 10(-9) M FeCl3), growth rates were not limited by photosynthetic capacity but rather by another, as of yet undetermined, iron-requiring cellular function . We have also investigated membrane proteins that are induced when the cells are grown in low iron medium . Using membrane fractionation techniques we were able to recognize specific proteins localized in the outer membrane and periplasmic space of O . tenuis . The recovery of growth rates at low iron levels occurred in parallel with the induction of these proteins and the production of extracellular siderophores . The additional iron acquired by this high affinity transport system did not reestablish photosynthesis in O . tenuis to the iron-satiated level but did reestablish growth to iron-replete levels . Oscillatoria tenuis appears to invoke an alternate physiology to compensate for iron deficiency. Appl Environ Microbiol, 1995 Dec, 61(12), 4487 - 9 Distribution and properties of fructosyl amino acid oxidase in fungi; Yoshida N et al.; Fructosyl amino acid oxidase, and enzyme that can be used for the determination of glycated proteins in blood samples from diabetic patients, was used to screen cultures in our microorganism culture collection . Fructosyl amino acid oxidase was found only in the strains of four genera of fungi, Aspergillus, Fusarium, Gibberella, and Penicillium and exhibited different substrate specificities against fructosyl valine and N epsilon-fructosyl N alpha-Z-lysine . A fructosyl valine-specific enzyme from Penicillium janthinellum AKU3413 was monomeric (M(r), 49,000), was most active at 35 degrees C and pH 8.0, and had a covalently bound flavin adenine dinucleotide as a prosthetic group. Am J Respir Crit Care Med, 1995 Dec, 152(6 Pt 1), 1982 - 91 Bronchoscopic or blind sampling techniques for the diagnosis of ventilator-associated pneumonia; Papazian L et al.; The purpose of this prospective postmortem study was to assess the diagnostic accuracy of bronchoscopic techniques (bronchoalveolar lavage {BAL} and protected specimen brush {PSB}) and nonbronchoscopic techniques (blind bronchial sampling {BBS} and mini-BAL) in the diagnosis of ventilator-associated pneumonia (VAP) . The results of each technique were compared with histology and culture of lung tissue specimens obtained by surgical pneumonectomies in 38 patients who died after at least 72 h of mechanical ventilation . Histology was positive for VAP in 18 patients and negative in 20 patients . There were 12 definite VAP (positive histology and positive lung cultures) and 6 histologic VAP (positive histology and negative cultures) . Clinical pulmonary infection score (CPIS) at a threshold of 6 achieved a sensitivity of 72% and a specificity of 85% . When the CPIS was combined with the logarithmic concentration of the predominant microorganism obtained from the BBS sample culture, specificity was increased to 95%, for a threshold of 10 . Using 10(3) cfu/ml as the threshold of positivity for cultures obtained with PSB and mini-BAL samples and 10(4) cfu/ml for cultures obtained with BBS and BAL, the respective sensitivities of these techniques for definite VAP were 42, 67, 83, and 58% . The sensitivity of BBS was significantly higher than that of PSB (p < 0.05) . The area under the receiver operator characteristic curve was significantly greater for BBS than PSB (p < 0.05) . Given that it is more sensitive and noninvasive, BBS is preferable to PSB for the diagnosis of VAP. J Nutr, 1995 Dec, 125(12), 3020 - 9 Whole body metabolism is not restricted to D-sugars because energy metabolism of L-sugars fits a computational model in rats; Livesey G et al.; Can L-sugars contribute to whole body energy metabolism? Energy balance studies were undertaken in rats fed L-sugars at a rate of 10 g/100 g basal diet . Partial metabolizable energy values (MEVp) during the last 28 of 56 d while consuming the L-sugar diets showed that L-glucose contributed no energy (0.3 +/- 0.9 kJ/g, P > 0.5, mean +/- SEM), whereas L-fructose and L-glucose contributed 6.9 +/- 0.9 (P < 0.001) and 8.8 +/- 1.8 (P < 0.001) kJ/g, respectively . Over periods from 0 to 28 and 0 to 56 d of L-sugar treatment, measurements were made of energy intake, the average lean mass of animals, fat and lean mass deposition . Using these measurements and a computational model, estimates were made of each supplement's net metabolizable energy value for maintenance (NEVs) . These estimates confirmed the supply of energy from L-fructose, with NEVs of 5.6 +/- 3.2 (P < 0.32) and 6.1 +/- 1.7 (P < 0.01) kJ/g over 28 and 56 d, respectively, and L-gulose with NEVs of 10.2 +/- 3.4 (P < 0.01) and 11.4 +/- 1.8 (P < 0.001) kJ/g over 28 and 56 d, respectively . A lack of energy supply from L-glucose was confirmed with NEVs of -2.6 +/- 3.2 (P > 0.5) and 0.0 +/- 1.6 (P > 0.5) kJ/g over 28 and 56 d, respectively . As reference substrates, sucrose and cellulose gave expected energy values in all determinations . Methods comparison analysis showed no differences between MEVp and NEVS . The assimilation of energy from L-fructose and L-gulose is probably via large bowl microorganisms . These observations show that L-gulose and L-fructose contribute to whole body energy metabolism while L-glucose does not. Med Clin (Barc), 1995 Nov 25, 105(18), 681 - 6 {Treatment and outcome of pneumococcal meningitis in adults . Study of a recent series of 70 episodes}; Almirante B et al.; BACKGROUND: Pneumococcal meningitis (PM) is an infection with high morbidity and mortality . The aim of this study was to evaluate the most relevant clinical, epidemiologic and evolutive characteristics of a recent series of adult patients with this disease . METHODS: Over a period of 10 years all the patients with PM diagnosed by isolation of this microorganism in the cerebrospinal fluid (CSF) were evaluated from a clinical, therapeutic and evolutive points of view . The impact of the new therapies in the disease and the variables associated with mortality were analyzed . RESULTS: Seventy episodes of PM were diagnosed, 60% being found in patients over the age of 50 years . The male/female relationship was 2/1 . Fifty-three percent of the patients had other underlying diseases . Acute otitis media (AOM) was the source in 34% of the cases, in 11% the patients had a fistula of CSF and in 9% a pneumonia . At the time of diagnosis 74% of the patients had some degree of reduction in the level of consciousness and in 40% of the episodes the presence of neurologic local manifestations were observed . A decrease in sensitivity to penicillin was observed in 33% of the microorganisms isolated . Third generation cephalosporins were used as initial treatment in 57 episodes and penicillin in other 11 episodes . Adjuvant treatment with dexamethasone, mannitol and/or diphenylhydantoin was administered in 54% of the patients . Overall mortality was 23%: the factors associated with an unfavourable evolution were the existence of underlying disease, deep alteration in the level of consciousness at the time of diagnosis, the coexistence of pneumonia and the absence of adjuvant therapy . CONCLUSIONS: Mortality in pneumococcal meningitis is high . The most relevant risk factor is the initial degree of consciousness . Adjuvant therapies probably determine a reduction in the rate of mortality. Structure, 1995 Nov 15, 3(11), 1207 - 15 Substrate binding and carboxylation by dethiobiotin synthetase--a kinetic and X-ray study; Alexeev D et al.; BACKGROUND: The vitamin biotin is a ubiquitous prosthetic group of carboxylase and transcarboxylase enzymes . Biotin biosynthesis occurs by similar pathways in microorganisms and plants . The penultimate step in biotin biosynthesis, catalyzed by dethiobiotin synthetase (DTBS), involves a unique ATP-dependent N-carboxylation, resulting in formation of the ureido ring function of dethiobiotin . The first two steps of dethiobiotin formation, which is a complex, multistep enzymatic reaction, have been elucidated by a combination of X-ray crystallography and kinetic methods . RESULTS: The first step in catalysis by DTBS is the formation of an enzyme-substrate complex and the second is the enzymatic carboxylation of the bound substrate . Both steps are Mg2+ dependent . The kinetic constants in the presence and absence of Mg2+ have been measured and a set of X-ray structures determined at different stages of the reaction . The conformational changes in the active site of the enzyme, induced by Mg2+, substrate binding and substrate carboxylation, have been monitored crystallographically and are discussed . Sulfate ions bound to DTBS may mimic the behaviour of the alpha- and gamma-phosphates of ATP in Mg2+ binding and in the subsequent steps of the reaction . CONCLUSIONS: Mg2+ is an essential cation for both substrate binding and carbamate formation by DTBS, when sulfate is present . The conformational changes induced at the active site in the DTBS-substrate complex, when Mg2+ is present, are small yet highly significant and serve to optimize the interactions between substrate and enzyme . DTBS is active as a homodimer and the substrate-binding site straddles both monomers in the dimer . The carboxylation site is unambiguously identified as the N-7 amino group of the substrate, rather than the N-8 amino group, as previously suggested . The elongated nucleotide-binding loop (the P loop) binds both ATP and substrate in a manner which suggests that this feature may be of wider importance. Rev Invest Clin, 1995 Nov-Dec, 47(6), 499 - 506 {Strategies of Histoplasma capsulatum for evading the cytocidal mechanisms of phagocytes}; Taylor ML et al.; Although proliferation of Histoplasma capsulatum within the macrophage is restricted by cell mediated immunity and, that there is no apparent deficiency in the fungicidal power of the macrophage, H . capsulatum can prosper in this intracellular microenvironment in some circumstances, which provides special nutritional advantages to intracellular growth for H . capsulatum, and access to other organs via the bloodstream and lymphatics . Nevertheless the environment within professional phagocytes is complex and generally hostile to microorganisms . Intracellular pathogens must avoid or overcome a series of obstacles in order to prevent their destruction . We present the strategies used by H . capsulatum to escape the host cell aggressiveness from the moment it reaches the cell surface up to its survival inside the phagocyte . Our aim is to discuss the advances on the escape mechanisms used by H . capsulatum to survive in the intracellular environment. Insect Mol Biol, 1995 Nov, 4(4), 237 - 43 PCR-based detection of Wolbachia, cytoplasmic incompatibility microorganisms, infected in natural populations of Laodelphax striatellus (Homoptera: Delphacidae) in central Japan: has the distribution of Wolbachia spread recently? Hoshizaki S, Shimada T. Cytoplasmic incompatibility is caused in various insects by intracellular infection with rickettsia-like microorganisms of the genus Wolbachia . In Japan Laodelphax striatellus shows unidirectional cytoplasmic incompatibility between northeastern and southwestern populations . In this study, nine natural populations of L . striatellus collected from central Japan, including the geographic boundary between the two cytotype populations, were analysed for Wolbachia infection by PCR using primers specific to Wolbachia 16S rDNA . The geographic pattern of the infection rates of the southwestern (high) and the northeastern (low or zero) populations broadly resembled that of a previous study of incompatibility . In populations which originated from the boundary regions between the southwestern and northeastern populations, the infected and uninfected cytotypes coexisted . It is suggested that in some populations of L . striatellus, which formerly had been uninfected with Wolbachia, the infection property has changed to the infected . Based on our results, we conclude that the distribution of Wolbachia-infected L . striatellus populations have spread northeasterly during the last 12 years. Histochem J, 1995 Nov, 27(11), 897 - 905 Ultrastructural localization of xanthine oxidase activity in the digestive tract of the rat; Van Den Munckhof RJ et al.; Precise localization of xanthine oxidase activity might elucidate physiological functions of the enzyme, which have not been established so far . Because xanthine oxidase is sensitive to chemical (aldehyde) fixation, we have localized its activity in unfixed cryostat sections of rat duodenum, oesophagus and tongue mounted on a semipermeable membrane . Previous studies had shown that this procedure enables the exact localization of activities of peroxisomal oxidases with maintenance of acceptable ultrastructure . Moreover, leakage and/or diffusion of enzyme molecules was prevented with this method . The incubation medium to detect xanthine oxidase activity contained hypoxanthine as substrate and cerium ions as capturing agent for hydrogen peroxide . After incubation, reaction product in the sections was either visualized for light microscopy or sections were fixed immediately and processed for electron microscopy . At the ultrastructural level, crystalline reaction product specifically formed by xanthine oxidase activity was found to be present in the cytoplasmic matrix of enterocytes and goblet cells and in mucus duodenum . Moderate activity was found in the cytoplasm of apical cell layers of epithelia of oesophagus and tongue, with highest activity in the cornified layer . Moreover, large amounts of reaction product were found to surround bacteria present between cell remnants of the cornified layer of the oesophagus . Many bacteria surrounded by the enzyme showed signs of destruction and/or cell death . The intracellular localization of xanthine oxidase activity in the cytoplasm of epithelial cells as well as the extracellular localization suggest that the enzyme plays a role in the lumen of the digestive tract, for instance in the defence against microorganisms. Zhonghua Kou Qiang Yi Xue Za Zhi, 1995 Nov, 30(6), 343 - 5, 384 {Production and characterization of specific monoclonal antibody against Porphyromonas endodontalis}; Xue Y et al.; Porphyromonas endodontalis was known to be important microorganisms in the etiology of pulp and apical infection . In this paper, we generated hybridomas secreting monoclonal antibody against Porphyromonas endodontalis ATCC 35406 . The specificity of the monoclonal antibody was examined by ELISA against a battery organisms (109 Strains) . The results indicated that the monoclonal antibody did not react with any non-Porphy romanas endodontalis (104 Strains) . So our monoclonal antibody is specific for Porphyromanas endodontalis and can be used in clinical samples for detection of pulp and apical infections. Gig Sanit, 1995 Nov-Dec, (6), 8 - 11 {The disinfection and preservation of drinking water with low-voltage pulsed electrical discharges}; Avchinnikov AV et al.; Two-hour exposure to low-voltage (2.8 to 3 kV) pulsed electric discharges (PED) effectively (by 99.9999%) disinfected drinking water . Preservation of water with IED had a pronounced bactericidal effect in respect of numerous pathogenic and opportunistic microorganisms . The aftereffect of PED persists for at least 2 months. Mikrobiologiia, 1995 Nov-Dec, 64(6), 833 - 44 {Bacterial processes of photosynthesis and dark assimilation of carbon dioxide in the lakes of Bunger Hills Oasis in Eastern Antarctica}; Gal'chenko VF et al.; Photosynthetic and dark assimilation of carbon dioxide was studied in both perennially and temporarily ice-covered lakes of the antarctic oasis Bunger Hills . The intensity of these processes in the lakes studied varied from 0.08 to 326 mg C/(m3 day) and correlated with water mineralization . Irrespective of the character of ice-cover, a chemical composition of water in antarctic water ecosystems affected the organic matter photo-production to a greater extent than the intensity of photosynthetically available radiation . With the method of polymerase chain reaction, the predominance of oxygenic phototrophic microorganisms carrying RuBisCo genes was revealed in all the lakes examined . The results obtained and the literature data unambiguously point to the photosynthetic production of organic matter by cyanobacteria as the main biogeochemical process that determines all the other metabolic strategies in antarctic water ecosystems studied hitherto. Mikrobiologiia, 1995 Nov-Dec, 64(6), 788 - 91 {Detection of methylotrophs in natural samples by amplifying a fragment of the moxF gene}; Beliaev AS et al.; A method for detection of methylotrophic microorganisms with the help of the polymerase chain reaction was developed . Two primers were synthesized, designed on the basis of comparing conservative sequences of the genes encoding the large subunit of methanol dehydrogenase . The developed system was tested with six strains of methylotrophic bacteria and with natural samples collected in the Bunger Hills oasis, eastern Antarctica . The results suggested a possibility of application of the developed method for qualitative detection of methylotrophs in natural samples. J Biochem Biophys Methods, 1995 Nov, 30(4), 257 - 69 Model components of luminol chemiluminescence generated by PMNL; Magrisso MJ et al.; The production of activated oxygen species (AOS) by neutrophils (PMNL) is thought to play a key role in the host defence against invading microorganisms . However, the oxygen metabolites are toxic not only to the invading bacteria but also to the surrounding tissue . The oxidative metabolites production can be evaluated by means of chemiluminescent methods . In this study, the possibility of a new analytical approach for quantitative assessment of chemiluminescent kinetics (AOS generation) of isolated PMNL was estimated . Based on the assumption that the kinetics of luminol-amplified chemiluminescence (LCL) of stimulated PMNL possesses a time-probabilistic nature, this kinetics was described with three components . These components, obtained from different investigated systems, were analyzed and a conclusion was made that the first and the second component represent the processes resulting in extra-and intracellular myeloperoxidase (MPO)-dependent light emission (AOS generation), respectively . The second component was found to be completely dependent on the stimulus ingestion . The third component was not completely MPO-dependent and complicated for interpretation . This component was weakly dependent on the stimulus ingestion, and presents at least some intracellular processes different from those presented by the second component . A conclusion is made that the examined approach for analysis of LCL kinetics allows an assessment of extra-and intracellularly generated quantities of AOS by stimulated PMNL . The assessment could be done for emitting systems in which no additional modifications are used. Chin Med J (Engl), 1995 Nov, 108(11), 839 - 43 MRI appearance of spinal lesions: metastatic tumors or infections? Thajeb P, Huang KM, Jeng CM, Shih CC, Lie SK, Huang JS. Three patients with pathologically verified spinal osteomyelitis and another three with metastatic tumors of the spine were investigated . MRI of th spine of four patients showed several unusual findings . The preservation of intervertebral discs and endplates did not predict accurately the diagnosis of either infections or tumors . The "pepper and salt" appearing feature may also occur in a metastatic tumor . Plain radiographs, CT, and radioisotope bone scans were less sensitive than MRI to disclose the lesions . In cases of osteomyelitis, the systemic sources of infections were frequently not found, and the responsible microorganisms could not be identified even from the surgical specimens of two patients . However, mycobacterium tuberculosis was found in the surgical specimen of a patient with cervical spinal lesion whose MRI was indistinguishable from a metastatic tumor . Surgicopathological diagnosis was therefore crucial and mandatory in these instances. Antimicrob Agents Chemother, 1995 Nov, 39(11), 2511 - 5 Methionine recycling pathways and antimalarial drug design; Sufrin JR et al.; 5'-Deoxy-5'-(methylthio)adenosine (MTA) is an S-adenosylmethionine metabolite that is generated as a by-product of polyamine biosynthesis . In mammalian cells, MTA undergoes a phosphorolytic cleavage catalyzed by MTA phosphorylase to produce adenine and 5-deoxy-5-(methylthio)ribose-1-phosphate (MTRP) . Adenine is utilized in purine salvage pathways, and MTRP is subsequently recycled to methionine . Whereas some microorganisms metabolize MTA to MTRP via MTA phosphorylase, others metabolize MTA to MTRP in two steps via initial cleavage by MTA nucleosidase to adenine and 5-deoxy-5-(methylthio)ribose (MTR) followed by conversion of MTR to MTRP by MTR kinase . In order to assess the extent to which these pathways may be operative in Plasmodium falciparum, we have examined a series of 5'-alkyl-substituted analogs of MTA and the related MTR analogs and compared their abilities to inhibit in vitro growth of this malarial parasite . The MTR analogs 5-deoxy-5-(ethylthio)ribose and 5-deoxy-5-(hydroxyethylthio)ribose were inactive at concentrations up to 1 mM, and 5-deoxy-5-(monofluoroethylthio)ribose was weakly active (50% inhibitory concentration = 700 microM) . In comparison, the MTA analogs, 5'-deoxy-5'-(ethylthio)adenosine,5'-deoxy-5'-(hydroxyethylthio)ade nosine (HETA), and 5'-deoxy-5'-(monofluoroethylthio)adenosine, had 50% inhibitory concentrations of 80, 46, and 61 microM, respectively . Extracts of P . falciparum were found to have substantial MTA phosphorylase activity . Coadministration of MTA with HETA partially protected the parasites against the growth-inhibitory effects of HETA . Results of this study indicate that P . falciparum has an active MTA phosphorylase that can be targeted by analogs of MTA. Am J Vet Res, 1995 Nov, 56(11), 1486 - 92 Pharmacokinetics of bacampicillin in equids; Sarasola P et al.; Bacampicillin hydrochloride is an ester prodrug that is hydrolyzed to ampicillin after its absorption from the gastrointestinal tract . It was administered intragastrically at a dose rate of 13.5 mg/kg of body weight to ponies and horses, and was highly bioavailable (F = 41.0%), compared with other penicillins in adult horses . The high peak ampicillin plasma concentration of 6.1 +/- 0.5 micrograms/ml achieved and persistence of the antibiotic at concentration of 0.3 +/- 0.1 microgram/ml 6 hours after its intragastric administration, suggest that bacampicillin hydrochloride may reach suitable bactericidal concentrations for treatment of infections caused by susceptible microorganisms . In a separate experiment, dichlorvos, an organophosphate compound that inhibits some of the esterase activity in plasma, was administered orally to the same animals at a dose rate of 40 mg/kg, followed by intragastric administration of bacampicillin hydrochloride at a dose of 13.5 mg/kg . Plasma pseudocholinesterase and erythrocyte acetylcholinesterase activities were reduced to < 5% of reference (predichlorvos) values after dichlorvos administration . However, rate of hydrolysis of bacampicillin into ampicillin was not affected . Consequently, the disposition and fate of bacampicillin when administered intragastrically 1 day after dichlorvos administration were similar to the values obtained after administration of bacampicillin alone . Intragastric coadministration of probenecid at a dose rate of 75 mg/kg and bacampicillin at 13.5 mg/kg limited absorption of the antibiotic from the gastrointestinal tract . This suggests existence of a common transport mechanism for bacampicillin and probenecid in the gastrointestinal wall, and precludes use of this combination for treatment . The bioavailable fraction of ampicillin after combination treatment indicated prolonged residence time in the plasma, presumably as a consequence of reduced renal tubular secretion. J Periodontol, 1995 Nov, 66(11), 978 - 83 Slow release of tetracycline hydrochloride from a cellulose membrane used in guided tissue regeneration; Markman C et al.; The objective of this study was to evaluate if the biologic membrane utilized for GTR can be impregnated by tetracycline hydrochloride and if the chemotherapeutic agent, once impregnated, can be released in minimal inhibitory concentrations for a period compatible with clinical application . Initially, an in vitro study was done with cellulose membranes cut in pieces measuring 9 cm2 . A volume of 100 microliters containing a 72,000 micrograms/ml solution of tetracycline was dispensed onto each fragment, and dried for 70 minutes at 37 degrees C . Four pieces measuring 0.5 cm2 were cut from different points of the 9 cm2 membrane (presumably, containing 400 micrograms of tetracycline), placed in test tubes containing 4 ml of sterile deionized water, and agitated for 2 minutes . A standard curve was made from known concentrations of tetracycline and compared to 10 microliters of the test solutions obtained by the elution of the 0.5 cm2 fragments . The concentrations were determined through the bioassay technique in 3 duplicate experiments . The samples recovered from the membrane fragments had a mean of 101 micrograms/ml of tetracycline liberated, demonstrating that the membrane was impregnated homogeneously by the chemotherapeutic agent . In a second phase, an in vivo study was carried out to determine the length of time the drug was liberated from the membranes and at which concentrations, in the presence of an inflammatory process . Fourteen 0.5 cm2 fragments containing 400 micrograms of tetracycline were placed in 14 polypropylene chambers containing 200 microliters of thioglycolate medium . The chambers were implanted in the peritoneal cavities of 14 mice, one chamber per animal, and left in from 1 to 14 days . They were then removed and the concentrations of tetracycline determined from 20 microliters samples using a bioassay . The results showed that the antibiotic was released slowly from the 1st through the 12th day in decreasing concentrations that varied from 218 to 20.8 micrograms/ml . The impregnated cellulose membrane can probably be used in GTR acting as a membrane and as a slow-release device, liberating the chemotherapeutic agent in concentrations high enough to eliminate periodontopathic microorganisms. J Antibiot (Tokyo), 1995 Nov, 48(11), 1320 - 9 Chemical modification of tazobactam . Synthesis of 2 beta-{(4-substituted)-1,2,3-triazol-1-yl}methyl penicillanic acid sulfone derivatives; Setti EL et al.; A series of 2 beta-{(4-substituted)-1,2,3-triazol-1-yl} methyl penicillanic acid sulfones was synthesized as beta-lactamase inhibitors . Many of these compounds showed good in vitro inhibitory activity against penicillinase, cefotaximase and plasmid-mediated class III TEM enzymes, but exhibited weaker cephalosporinase inhibition . One member in this series--2 beta-{(4-pyridiniummethyl)-1,2,3-triazol- 1-yl}-6,6-dihydropenicillanate 1,1-dioxide (12a), when tested in combination with piperacillin, showed excellent synergistic activity against microorganisms producing plasmid-mediated enzymes, but had insufficient activity against microorganisms producing chromosomally mediated class I enzymes. J Clin Periodontol, 1995 Nov, 22(11), 842 - 9 Antibody responses to suspected periodontal pathogens in elderly subjects with periodontal disease; McArthur WP et al.; Little is known about the relationship of aging to periodontal disease . The immune response undergoes aging-related changes resulting in loss of functional capacity . The aim of this study was to investigate the relationship between levels of serum IgG antibodies against suspected periodontal pathogenic microorganisms to the presence or absence of periodontal disease in an elderly (65-75 yrs) population . From this study, we obtained information concerning: (1) the ability to differentiate elderly individuals without disease from those with disease by their levels of antibodies against periodontal pathogens and (2) which periodontal pathogen(s) triggered those responses . IgG anti- Porphyromonas gingivalis (strains W83 and 381) levels in the serum of elderly patients with severe periodontal disease were the only antibody responses measured which were elevated compared to the elderly control group of subjects with no periodontal disease . Anti- Prevotella intermedia IgG levels in both elderly patient groups were depressed compared to anti- P . intermedia levels in the young normal control subjects . Serum IgG antibody levels to six other plaque microorganisms did not differentiate between diseased and normal, elderly or young subjects . This data suggested that P . gingivalis was associated with periodontal disease in this elderly group of individuals and that those elderly individuals were able to respond with a normal IgG immune response. Bone Marrow Transplant, 1995 Nov, 16(5), 675 - 81 Autopsy-identified infections among bone marrow transplant recipients: a clinico-pathologic study of 56 patients . Bone Marrow Transplantation Team; Chandrasekar PH et al.; We reviewed the autopsy records of 56 patients who had undergone BMT at the Detroit Medical Center during 1988-1992 . Most patients (43 of 56) had died within 2 months of BMT . One or more infections were identified at autopsy in 25 of 40 (63%) allogeneic and four of 16 (25%) autologous BMT recipients . Microorganisms isolated at autopsy were cytomegalovirus (CMV) (14 patients), yeasts (13 patients), molds (aspergillus six patients, mucor one patient) and bacteria (seven patients) . Presence of infection was not identified or proven prior to death in nine of 14 patients (65%) with CMV, six of 13 patients (46%) with yeasts and four of six patients (67%) with aspergillus . Most bacterial infections (five of seven patients) were identified ante-mortem . Lungs and the gastrointestinal tract were the organ systems mostly involved in patients with or without autopsy-identified infections . Pathologic findings in the lungs were diffuse alveolar damage, interstitial pneumonia and bronchopneumonia and, in the gastrointestinal tract, were ulcerations and hemorrhages of esophagus, stomach, small and large intestines . Examination of the heart showed non-bacterial thrombotic endocarditis (NBTE) in five patients, four with right-sided involvement only . Failure to identify non-bacterial pathogens ante-mortem and their frequent association with mortality in bone marrow transplant patients are high-lighted in the present study. Clin Exp Immunol, 1995 Nov, 102(2), 231 - 7 Accumulation of activated CD4+ lymphocytes in the lung of individuals infected with HIV accompanied by increased virus production in patients with secondary infections; Franchini M et al.; The lung is continuously exposed to infectious and non-infectious agents causing cell activation . Activated cells in the lung such as antigen-presenting cells which harbour HIV may favour this organ as a site for virus production . To test this hypothesis, cells from blood and bronchoalveolar lavage (BAL) of HIV-infected patients and healthy controls were obtained and the activation of the cells were analysed by measuring the expression of IL-2 receptor, HLA-DR and VLA-1 . The HIV-infected individuals were subdivided into 'lung symptomatic' or 'lung asymptomatic' patients, depending on the presence or absence of secondary lung diseases besides HIV . All HIV-infected individuals demonstrated a decreased number of CD4+ lymphocytes in blood; however, normal numbers of these cells were found in BAL . The activation state of CD4+ and CD8+ T lymphocytes in blood and BAL was higher in lymphocytes from HIV-infected patients compared with controls . The activation state was highest in the lung symptomatic group . Lung symptomatic patients and lung asymptomatic patients with extrapulmonary infections had increased levels of free virus in plasma . Four out of four individuals without or with only low amounts of cell-free HIV in plasma belonged to the symptom-free subgroup . These results suggest that microorganisms other than HIV may promote viral replication via antigen-driven accumulation and activation of CD4+ cells in the lung or other organs, and thus may be responsible for the loss of helper T cells and the progression of the disease. Am J Obstet Gynecol, 1995 Nov, 173(5), 1527 - 31 Intravaginal clindamycin treatment for bacterial vaginosis: effects on preterm delivery and low birth weight; Joesoef MR et al.; OBJECTIVE: Our goal was to evaluate whether treatment of bacterial vaginosis during pregnancy with 2% clindamycin vaginal cream reduces the incidence of either preterm delivery or low birth weight or of both . STUDY DESIGN: A multicenter, double-blind, randomized, placebo-controlled trial in Indonesia compared a 2% clindamycin vaginal cream with a placebo cream . Women seeking prenatal care at 14 to 26 weeks of gestational age who had bacterial vaginosis (Gram stain score > 6 and pH of vaginal fluid > 4.5) were invited to participate . Of the 745 women enrolled, 681 (91.4%) women were followed up through delivery . RESULTS: Clindamycin vaginal cream was an effective treatment for bacterial vaginosis . Two weeks after completion of the treatment, 85.5% of the women were cured . The rate of preterm delivery (< 37 weeks) was 15.0% for clindamycin patients and 13.5% for placebo patients (odds ratio 1.1, 95% confidence interval 0.7 to 1.7) . The rate of low birth weight was 9.0% for clindamycin patients and 6.8% for placebo patients (odds ratio 1.3, 95% confidence interval 0.8 to 2.4) . CONCLUSIONS: Treatment of bacterial vaginosis with clindamycin vaginal cream did not reduce preterm delivery or low birth weight . Although clindamycin vaginal cream is an effective treatment for bacterial vaginosis, intravaginal treatment would not be effective against bacterial vaginosis-associated microorganisms harbored in the upper genital tract . Systemic treatment may be required to eradicate upper tract infection to reduce preterm delivery. Tidsskr Nor Laegeforen, 1995 Oct 30, 115(26), 3259 - 62 {New immunopathogenetic aspects of infectious diseases}; Muller F et al.; The clinical course of infectious diseases depends on the complicated interaction between the microorganism and the immune system of the host . The vast amount of knowledge acquired during recent years in the field of immunology will probably lead to new ways of treating infectious diseases by selective immunomodulation . In this paper, we review some newer aspects of host defence mechanisms of relevance to immunomodulation of infectious diseases . The cytokine network, apoptosis, superantigens, and oxidative stress are all discussed with reference to various infectious diseases . Furthermore, the possible role of physical and psychological stress in the immunopathogenesis of infectious disease is discussed in the light of the interplay between the immune system and the neuroendocrine system. FEBS Lett, 1995 Oct 16, 373(3), 212 - 6 Alcohol dehydrogenase of class III: consistent patterns of structural and functional conservation in relation to class I and other proteins; Hjelmqvist L et al.; Class III alcohol dehydrogenase from the lizard Uromastix hardwickii has been characterized . This non-mammalian, gnathostomatous vertebrate class III form allows correlations of structures and functions of this class, the traditional class I alcohol dehydrogenase, and other well-studied proteins . Catalytically, results show similar recoveries and activities of all vertebrate class III forms independent of source, similar activities also in invertebrates but in lower amounts, and considerably higher specific activities in microorganisms . Structurally, variability patterns are consistent throughout the vertebrate system with a ratio in accepted point mutations versus class I of 0.4 . This ratio between different classes of a zinc enzyme is comparable to that between different heme proteins (cytochrome c and myoglobin), suggesting defined but non-identical functions also for the alcohol dehydrogenase classes. Am J Vet Res, 1995 Oct, 56(10), 1390 - 5 Ruminal microbial alterations associated with sulfide generation in steers with dietary sulfate-induced polioencephalomalacia; Cummings BA et al.; Holstein steers were fed carbohydrate-rich, short-fiber basal diets with and without added sodium sulfate . Steers fed the high-sulfate diet developed the CNS disorder polioencephalomalacia (PEM) . The onset of signs of PEM was associated with increased sulfide concentration in the rumen fluid . Over the course of the disease, anaerobic rumen bacteria were enumerated in roll tubes by use of the Hungate method Lo determine the effect of dietary sulfate on sulfate-reducing bacterial numbers . Media used included a general type for total counts and sulfate containing media with and without cysteine to assess sulfate-reducing bacteria . Changes in total and sulfate reducing bacterial numbers attributable to dietary sulfate content were not observed . The capacity to generate hydrogen sulfide from sulfate in fresh rumen fluid in vitro was substantially increased only after steers had been fed the high sulfate diet for 10 to 12 days, which coincided with the onset of signs of PEM . The low capacity for hydrogen sulfide production of rumen fluid taken at earlier times in the feeding period suggests that rumen microorganisms must adapt to higher dietary sulfate content before they are capable of generating potentially toxic concentrations of sulfide. Singapore Med J, 1995 Oct, 36(5), 539 - 44 Community-acquired pneumonia; Ong KC et al.; Major changes have occurred in the epidemiology of community-acquired pneumonia recently . The emergence of new pathogens emphasises the need for continued vigilance in the diagnosis of pneumonia while changes in the microorganism or in the host have resulted in exciting new aspects of several old pathogens . Clinical and radiologic signs are unreliable in predicting the infecting organisms . Thus initial therapy is nearly always empiric . This approach often requires good clinical judgement and a knowledge of local epidemiological patterns in choosing an appropriate regimen . State-of-the-art invasive diagnostic procedures are usually reserved for pneumoniae that fail to resolve with initial treatment . Non-specific measures like stabilisation of underlying medical conditions, adequate nutrition and cessation of smoking or alcohol may help prevent the development of community-acquired pneumonia . On a larger scale, influenza and pneumococcal vaccinations are cost-effective preventive measures. Eur J Clin Microbiol Infect Dis, 1995 Oct, 14(10), 839 - 50 The role of bronchoalveolar lavage in the diagnosis of bacterial pneumonia; Sanchez Nieto JM et al.; Bronchoalveolar lavage (BAL) has become an invaluable diagnostic tool with important clinical implications in both opportunistic infections and the pulmonary pathology of immunologic disease . Until recently, the use of BAL was limited primarily to two areas: the study of interstitial lung diseases and the diagnosis of lung infections by opportunistic microorganisms in severely immunocompromised patients with lung infiltrates . Over the past decade, the use of BAL has been expanded to include the conventional diagnosis of bacterial pneumonia in non-immunocompromised patients . In the past, different clinical studies proposed using BAL to quantify cultures in the sample obtained as a means of increasing the tool's effectiveness . Recent developments have led to a number of newer applications of BAL, such as bronchoscopic BAL, non-bronchoscopic BAL and protected BAL . The most important use of BAL in the non-immunocompromised patient is the diagnosis of pneumonia in the mechanically ventilated patient. J Ind Microbiol, 1995 Oct, 15(4), 352 - 60 Biosynthesis of the polysialic acid capsule in Escherichia coli K1; Vimr E et al.; The extracellular polysaccharides elaborated by most or all bacterial species function in cell-to-cell and cell-substratum adhesion, cell signaling, and avoidance or inhibition of noxious agents in animal hosts or free-living environments . Recent advances in our understanding of exopolysaccharide synthesis have been facilitated by comparative approaches in both plant and animal pathogens, as well as in microorganisms of industrial importance . One of the best understood of these systems is the kps locus for polysialic acid synthesis in Escherichia coli K1 . The genes for sialic acid synthesis, activation, polymerization and translocation have been identified and assigned at least tentative functions in the synthetic and export pathways . Initial studies of kps thermoregulation suggest that genetic control mechanisms will be involved which are distinct from those already described for several other exopolysaccharides . Information about the common as well as unique features of polysialic acid biosynthesis will increase our knowledge of microbial cell surfaces which in turn may suggest novel targets for therapeutic or industrial interventions. Compend Contin Educ Dent . 1995 Oct;16(10):1042, 1044, 1046 passim; quiz 1058. The importance of periodontal pathogens in guided periodontal tissue regeneration and guided bone regeneration; Nowzari H et al.; Although guided tissue regeneration (GTR) procedures in periodontitis lesions and around endosseous dental implants represent exciting new therapeutic modalities in periodontics, these treatments can fail because of shortcomings in surgical techniques, restriction in the size and shape of the defect, anatomic features interfering with surgery, or infectious complications . Our studies show that optimal tissue regeneration cannot be expected for a nonbioabsorbable barrier membrane placed in a site infected by periodontopathic microorganisms . Our data also indicate that treatment failure is most frequent in patients who harbor high levels of periodontal pathogens and show evidence of severe periodontitis in numerous teeth . To decrease the risk of infection and to ensure proper healing, periodontal therapy should precede insertion of the barrier membrane for GTR . Recently, we have studied the effect of the pathogens on periodontal GTR and guided bone regeneration around dental implants and the results are reviewed in this article. J Dairy Sci, 1995 Oct, 78(10), 2230 - 8 Validity of using accumulated gas pressure readings to measure forage digestion in vitro: a comparison involving three forages; Schofield P et al.; Gas output from in vitro digestion of forage can be used to measure both forage digestibility and the kinetics of microbial digestion . Two different approaches are used to measure gas volumes: 1) either the gas is collected at atmospheric pressure and its volume determined directly, or 2) the gas accumulates in a fixed volume container, and the volume is calculated from pressure changes . We have used the latter technique to study the in vitro digestion of alfalfa, timothy, and corn silage by ruminal microorganisms in bottles of different sizes . We measured pH changes, NDF disappearance, and VFA concentrations and characterized the digestion profiles using a single- or dual-pool logistic model . Bottle size had a slight effect on pH but was not associated with changes in any of the other characteristics measured including the kinetic parameters . Gas measurements based on pressure increases in a fixed volume container appear to offer a valid alternative to methods based on gas collection at atmospheric pressure . We present an equation to adjust pressure sensor calibration values for use in different sized containers. Can J Microbiol, 1995 Oct, 41(10), 862 - 8 Study of the reductive dechlorination of pentachlorophenol by a methanogenic consortium; Juteau P et al.; Pentachlorophenol (PCP) dechlorination by a methanogenic consortium was observed when glucose, formate, lactate, or yeast extract was present in the mineral medium as a secondary carbon source . Acetate was not a good substrate to sustain dechlorination . The consortium was able to dechlorinate the different monochlorophenols, although the chlorine in position ortho and meta was removed more readily than in para position . Dechlorination was most efficient at 37 degrees C . At 45 degrees C, the first PCP dechlorination steps were very rapid, but 3,5-dichlorophenol (3,5-DCP) was not further dechlorinated . At 15 and 4 degrees C, dechlorination was very slow . The dechlorination of PCP to 3-chlorophenol (3-CP) was still observed after the consortium had been subjected to heat treatment (80 degrees C, 60 min), suggesting that spore-forming bacteria were responsible . The dechlorinating activity of the consortium was significantly reduced by the presence of hydrogen, 2-bromoethanosulfonic acid (BESA), or sulfate but not of nitrate . The dechlorination of 3-CP was completely inhibited by heat treatment or the presence of BESA, suggesting that a syntrophic microorganism would be involved . Vigorous agitation of the consortium stopped the dechlorination, but the presence of DEAE-Sephacel acting as a support was very efficient in restoring the activity, suggesting that association between certain members of the consortium was important. New Microbiol, 1995 Oct, 18(4), 399 - 407 Monoclonal antibodies against antigens exposed on the surface of vegetative forms and spores of Myxococcus virescens; Martinelli F et al.; Twelve monoclonal antibodies (mAbs) directed against cell-surface antigens of Myxococcus virescens cells were developed and partially characterized . All of them recognized multiple, diffuse proteic bands in Western blot and four were also reactive to living bacteria, as assessed by flow cytometry . The four latter mAbs recognized antigens common to a number of vegetative forms and spores . The selective expression of proteins recognized by mAbs on the microorganisms and the possible applications of mAbs to the study of myxobacterial cell interaction are discussed. Klin Monatsbl Augenheilkd, 1995 Oct, 207(4), 253 - 63 {Surface quality of flexible silicone intraocular lenses . A scanning electron microscopy study}; Kohnen T et al.; BACKGROUND: Soft intraocular lenses (IOLs) have been developed to be folded during insertion to allow implantation through a small incision . The surface of the IOL is of great importance in postoperative inflammation and long-term acceptance of the implant . Rough and sharp edges can damage delicate intraocular tissues . The purpose of this study was to analyze new, foldable silicone IOLs for surface quality prior to and following folding . MATERIALS AND METHODS: Eleven silicone IOLs of different types were included in this study (four one-piece plate-haptic silicone IOLs and seven three-piece silicone IOLs with polypropylene, PMMA or polyimid haptics) . We performed scanning electron microscopy on brand-new IOLs prior to and following folding either with forceps or inserter . Special attention was given to the silicone optic surface, optic edges, haptic-optic junctions and the haptic itself . Photographs were taken at 5-350 times magnification . RESULTS: All IOLS demonstrated a smooth and homogeneous optic surface at low magnification . At high-power magnification (X 350), distinctive surface patterns were evident in some IOLs, which turned out to be artefacts . The edge finish showed surplus silicone material and molding flash in six of 11 IOLs . Positioning holes of the 4 plate-haptic IOLs were, except in one IOL, rounded and not rough . Photographs of the haptic-optic junctions revealed surplus material or clefts between the haptic and optic in six of the 7 three-piece IOLs; the loop ends of two IOLs showed a roughened or irregular surface . We did not detect any IOL changes produced by folding . CONCLUSIONS: The silicone IOLs tested demonstrated generally acceptable surface properties, but most IOLs had regional surface irregularities of varying magnitude . The clinical impact of these remains to be established, but surplus material or surface defects might result in deposition of inflammatory cells, protein or microorganisms and synechia formation . Folding of the IOLs did not produce superficial defects. Res Microbiol, 1995 Oct, 146(8), 697 - 706 High pressure influences on gene and protein expression; Bartlett DH et al.; Elevated hydrostatic pressure can influence gene and protein expression in both 1 atmosphere-adapted and high pressure-adapted microorganisms . Here we review experiments documenting these effects and describe their significance towards understanding the molecular bases of life in deep-sea high pressure environments. J Vet Diagn Invest, 1995 Oct, 7(4), 465 - 72 Detection of Anaplasma marginale DNA in bovine erythrocytes by slot-blot and in situ hybridization with a PCR-mediated digoxigenin-labeled DNA probe; Ge NL et al.; A 409-base pair (bp) DNA fragment derived from the msp-1 beta gene of Anaplasma marginale was amplified and simultaneously labeled with digoxigenin-11-dUTP by a polymerase chain reaction (PCR) assay . The resulting digoxigenin-labeled 409-bp PCR product was used as a probe for slot-blot and in situ hybridization to detect A . marginale DNA from experimentally infected bovine erythrocytes . The hybrid formation was detected with alkaline phosphatase-conjugated anti-digoxigenin antibody and substrates 5-bromo-4-chloro-3-indolyl phosphate and nitroblue tetrazolium salt . In slot-blot hybridizations, the probe detected A . marginale DNA from approximately 1,000-10,000 infected erythrocytes in 1.25 ml of whole blood, which is equivalent to a parasitemia level of 0.00001% . The probe proved to be a A . marginale-specific when tested with 17 species of microorganisms . The applicability of the probe for diagnosis was tested by screening A . marginale infections in 2 experimentally infected splenectomized cattle before microscopically detectable parasitemias and after acute infection . After inoculation of infected blood, A . marginale infections were detected with the probe 14 days prior to detection in stained smears . Microscopically inapparent parasitemias were also detected with the probe for 2 months after acute disease . When the probe was used for in situ hybridization on methanol-fixed blood smears, probe reaction could be visualized with light microscopy on A . marginale inclusions within infected erythrocytes . The probe reaction was not observed on leukocytes and uninfected erythrocytes from infected blood smears, on erythrocytes from uninfected blood samples, or on samples infected with A . ovis, Babesia bovis, or B . bigemina . This PCR-mediated nonradioactive probe appears to be a sensitive diagnostic test for A . marginale. J Clin Microbiol, 1995 Oct, 33(10), 2710 - 4 Hemocytic rickettsia-like organisms in ticks: serologic reactivity with antisera to Ehrlichiae and detection of DNA of agent of human granulocytic ehrlichiosis by PCR; Magnarelli LA et al.; Ixodid ticks were collected from Connecticut, Massachusetts, Missouri, Pennsylvania, Rhode Island, and British Columbia (Canada) during 1991 to 1994 to determine the prevalence of infection with hemocytic (blood cell), rickettsia-like organisms . Hemolymph obtained from these ticks was analyzed by direct and indirect fluorescent antibody (FA) staining methods with dog, horse, or human sera containing antibodies to Ehrlichia canis, Ehrlichia equi, or Rickettsia rickettsii . Of the 693 nymphal and adult Amblyomma americanum, Dermacentor variabilis, Ixodes scapularis, and Ixodes pacificus ticks tested with dog anti-E . canis antiserum, 209 (32.5%) contained hemocytic bacteria . The prevalence of infected ticks varied greatly with species and locale . In parallel tests of duplicate hemolymph preparations from adult I . scapularis ticks, the hemocytic organisms reacted positively with E . canis and/or E . equi antisera, including sera from persons who had granulocytic ehrlichiosis . In separate PCR analyses, DNA of the agent of human granulocytic ehrlichiosis was detected in 59 (50.0%) of 118 adult and in 1 of 2 nymphal I . scapularis ticks tested from Connecticut . There was no evidence of Ehrlichia chaffeensis DNA in these ticks . In indirect FA tests of hemolymph for spotted fever group rickettsiae, the overall prevalence of infection was less than 4% . Specificity tests of antigens and antisera used in these studies revealed no cross-reactivity between E . canis and E . equi or between any of the ehrlichial reagents and those of R . rickettsii . The geographic distribution of hemocytic microorganisms with shared antigens to Ehrlichia species or spotted fever group rickettsiae is widespread. J Interferon Cytokine Res, 1995 Oct, 15(10), 837 - 8 Effective treatment with interferon-alpha in chronic recurrent multifocal osteomyelitis; Andersson R; Chronic recurrent multifocal osteomyelitis (CRMO) is a rare disease of unknown etiology characterized by multiple osteomyelitic changes in the predominantly metaphysial regions of long bones . It was first described by Giedon et al . in 1972 . Cultures for all known microorganisms are negative . Pain is the most common symptom, and sometimes soft tissue swelling is present . Patients are usually treated with nonsteroidal antiinflammatory drugs (NSAIDs) or corticosteroids and respond, at least partly, to these treatments . CRMO is most commonly seen in children and is in the majority of cases self-limiting but has a protracted course of several years . Some patients have a more prolonged disease period, as in the patient reported here . Treatment with corticosteroids in children has the risk of causing growth retardation as a potential adverse effect, and alternative treatments are of great interest . In the actual paper, a successful treatment with interferon-alpha 2b in a 34-year-old man with CRMO is presented. Lipids, 1995 Oct, 30(10), 899 - 906 A temperature-sensitive strain of Histoplasma capsulatum has an altered delta 9-fatty acid desaturase gene; Gargano S et al.; We have isolated and characterized the delta 9-desaturase gene (Ole1), which codes for a key enzyme involved in regulating membrane fluidity in animal cells and microorganisms, from two strains of Histoplasma capsulatum, one that is temperature-tolerant (G217B) and the other temperature-susceptible (Downs) . These pathogenic fungi are dimorphic in that they undergo a morphologic transition from the mycelial to yeast-like form when the temperature of incubation is switched from 25 to 37 degrees C or when they infect a susceptible host . The coding sequences of the two genes, both containing an intron of 93 nucleotides, are virtually identical and analogous to the delta 9-desaturase gene of Saccharomyces cerevisiae and those of the rat, mouse and human . Ole1 transcription of the thermotolerant G217B and thermosensitive Downs strains is similar in yeast phase cells and during the temperature shift down from 34, 37, or 40 to 25 degrees C (yeast-to-mycelia transition) . Nevertheless, the delta 9-desaturase gene is transcriptionally inactive in mycelia of G217B at 25 degrees C while it is actively transcribed in the Downs strain at the same temperature . These results are in agreement with the finding that membranes of the Downs strain have a higher level of oleic acid . The differential expression of delta 9-desaturase genes is discussed in relationship to differences in thermosensitivity in the fungal isolates and in regulating the level of expression of heat shock genes.