Genitourin Med, 1993 Oct, 69(5), 400 - 3 Clinical features and management of recurrent balanitis; association with atopy and genital washing; Birley HD et al.; OBJECTIVE--To evaluate clinical features and diagnostic investigations in patients with recurrent or unresponsive balanitis in order to institute rational management . DESIGN--Forty-three patients presenting to a genitourinary medicine clinic with recurrent or persistent balanitis were studied . All patients were asked whether they had a history of atopic illness and about their practice of genital washing . All patients were investigated by taking a swab specimen from the preputial area for bacterial and viral culture and 30 underwent biopsy of the affected skin . Follow-up was between three and six months . SETTING--Outpatient genitourinary medicine clinic, St Mary's Hospital, London, UK . RESULTS--In 31 (72%) of the patients a diagnosis of irritant dermatitis was made . In comparison with the remaining patients, they had a greater lifetime incidence of atopic illness and more frequent daily genital washing with soap . For 28 (90%) of these patients, use of emollient creams and restriction of soap washing alone controlled symptoms satisfactorily . For the remaining 12 patients, a variety of diagnoses were made . Biopsy proved a well tolerated and diagnostic investigation, but the isolation of microbial pathogens from preputial swabs was irrelevant to management . CONCLUSION--A history of atopic illness and of the practice of penile washing are important aspects in the evaluation of patients with recurrent balanitis . Biopsy is an important investigation in the condition when it does not seem to be caused by irritant dermatitis. Am J Obstet Gynecol, 1993 Oct, 169(4), 764 - 74 Antibiotic treatment of preterm labor with intact membranes: a multicenter, randomized, double-blinded, placebo-controlled trial; Romero R et al.; OBJECTIVE: Although an association between subclinical intrauterine infection and preterm birth is well established, there is conflicting evidence regarding the benefits of antibiotic administration to women in preterm labor with intact membranes . We attempted to determine the effect of ampicillin-amoxicillin and erythromycin treatment on prolongation of pregnancy, the rate of preterm birth, and neonatal morbidity in patients with preterm labor and intact membranes . STUDY DESIGN: A multicenter, randomized, double-blinded, placebo-controlled trial was designed and implemented by the Maternal-Fetal Medicine Units Network of the National Institute of Child Health and Human Development . Two hundred seventy-seven women with singleton pregnancies and preterm labor with intact membranes (24 to 34 weeks) were randomly allocated to receive either antibiotics or placebos . RESULTS: Of the 2373 patients screened for participation in this study in six medical centers, 277 women were enrolled (n = 133 for antibiotics group vs n = 144 for placebo group) . In each study group, 60% of patients completed all the study medications . The overall prevalence of microbial invasion of the amniotic cavity was 5.8% (14/239) . No significant difference between the antibiotic group and the placebo group was found in maternal outcomes, including duration of randomization-to-delivery interval, frequency of preterm delivery (< 37 weeks), frequency of preterm premature rupture of membranes, clinical chorioamnionitis, endometritis, and number of subsequent admissions for preterm labor . Similarly, no significant difference in neonatal outcomes could be detected between the two groups including respiratory distress syndrome, bronchopulmonary dysplasia, intraventricular hemorrhage, sepsis, and admission and duration of newborn intensive special care unit hospitalization . CONCLUSION: The results of this study do not support the routine use of antibiotic administration to women in preterm labor with intact membranes. J Anim Sci, 1993 Oct, 71(10), 2777 - 84 Metabolizable protein and amino acid requirements of growing cattle; Wilkerson VA et al.; Metabolizable protein and amino acid requirements for growing cattle were estimated using data from 11 research trials . A total of 543 steers were individually fed a high-roughage diet supplemented with protein at several levels above a urea supplement control . The mean weight for all animals was 253 kg, with a range in mean initial to final weights of 200 to 316 kg, respectively . Daily gain ranged from -.04 to .89 kg . Metabolizable protein for each treatment group was calculated at the point at which the protein requirement was met . The sum of dietary escape protein (basal and supplemental) and calculated microbial protein represented metabolizable protein supplied per test protein source analyzed in each trial . Daily gain was regressed against calculated metabolizable protein flow using weighted regression analysis (r2 = .69, n = 45) to determine the metabolizable protein requirements for maintenance (3.8 x BW.75 g/d, where BW is expressed in kilograms) and growth (305 g/kg of live weight gain) . Calculated metabolizable amino acid requirements as a percentage of metabolizable protein for a 253-kg animal gaining .49 kg/d were as follows: methionine, 3.0%; total sulfur amino acids, 5.8%; lysine, 8.0%; tryptophan, 1.0%; threonine, 5.2%; valine, 5.7%; isoleucine, 5.6%; leucine, 6.9%; phenylalanine, 3.9%; and histidine, 1.6% . The proposed requirements were based on live animal gain and intake of metabolizable protein and should represent the needs of the growing beef animal. Ann Hematol, 1993 Oct, 67(4), 191 - 6 The acute infection-associated hemolytic anemia of childhood: immunofluorescent detection of microbial antigens altering the erythrocyte membrane; Seitz RC et al.; The majority of acute infection-associated hemolytic diseases of infancy and childhood have been suggested to be caused by exogenic alterations of the erythrocyte surface, though laboratory methods for their further evaluation were not yet available . Investigating 96 children, the present study characterizes 72% of cases as corresponding to this type of acute acquired hemolytic anemia, which cannot be clearly related to autoantibodies against unmodified components of the host's own red cells . Using a new immunofluorescence test, the erythrocyte membrane of 80% of these children was found to be altered in vivo by nonspecific adsorption of foreign material released from the infectious micro-organisms . In 24% of cases additive binding of complement was detectable by an antiglobulin test . Thus, the adsorption of microbial antigens to the red cell surface is suggested to be one of the causes for the removal of altered erythrocytes due to phagocytosis or a complement-dependent destruction during the course of infection-associated hemolytic anemia . Especially in childhood, the immunofluorescent detection of an erythrocyte sensitization in vivo provides a further characterization of this type of mostly transient hemolytic disease, which probably can be treated without any immunosuppressive drug, merely by elimination of the underlying infection. Mol Cell Biol, 1993 Oct, 13(10), 5970 - 80 The erbA oncogene represses the actions of both retinoid X and retinoid A receptors but does so by distinct mechanisms; Chen HW et al.; Genetic lesions that function as dominant negative mutations in microbial systems have long been recognized . It is only relatively recently, however, that similar dominant negative mutations have been implicated as a basis for genetic and neoplastic disorders in vertebrates . We describe here a dissection of the actions of the erbA oncogene protein, an aberrant form of thyroid hormone receptor that acts as a dominant negative inhibitor of other nuclear hormone receptors . We demonstrate that the ErbA oncoprotein interferes with thyroid hormone and trans-retinoic acid receptors by competing for binding to the corresponding response elements . Heterodimerization of the ErbA oncoprotein with these receptors does not play an observable role in repression . In contrast, however, the ErbA oncoprotein does efficiently form a heterodimer with the retinoid X receptor (RXR) class of nuclear hormone receptors; complex formation enhances the DNA-binding properties of the ErbA protein but dramatically interferes with the ability of the RXR component to activate gene expression . Our results indicate that the erbA oncogene may play a previously unanticipated role in neoplasia by interfering with RXR function. Biosci Biotechnol Biochem, 1993 Oct, 57(10), 1628 - 31 Effects of microbial products on glucose consumption and morphology of macrophages; Magae J et al.; We studied the effects of microbial products on glucose consumption and morphology of macrophages which were elicited with thioglycollate medium . Macromolecules such as lipopolysaccharide (LPS), tumor promoters, and respiratory inhibitors increased macrophage glucose consumption without inducing evident morphological changes . The assay system was used to screen for active substances in culture broth extracts from actinomycetes . Among them, aureothin increased glucose consumption of macrophages and inhibited respiration of a rat mitochondrial fraction . Concanamycin A induced morphological changes of macrophages into needle-like shapes but not of cloned cells including the macrophage-like cells J774.1 . This compound changed fibrosarcoma L929 cells into round shapes without affecting the shape of a nontransformed fibroblast, BALB/3T3 cells . Antimycin and concanamycin A increased tumor-killing activity of macrophages when added during the effector phase . These results suggest that this assay system is simple and sufficiently reproducible and thus usable for screening for modulators of macrophage function among natural products. Curr Opin Immunol, 1993 Oct, 5(5), 763 - 73 Cyclosporin A and FK506: molecular mechanisms of immunosuppression and probes for transplantation biology; Bierer BE et al.; The microbial products cyclosporin A (CsA), FK506 and rapamycin are potent immunosuppressive agents . The introduction of CsA in the early 1970's significantly improved the outcome of organ and bone marrow allograft transplantation and advanced therapeutic options in autoimmune diseases . FK506 appears to have a higher therapeutic index than CsA, and has been used with encouraging results in clinical transplantation trials . FK506 and CsA, although structurally unrelated, appear to target similar signal transduction pathways in hematopoietic cells by inhibiting the action of calcineurin, a serine/threonine phosphatase . A structural analog of FK506, rapamycin, inhibits cellular function by a different molecular mechanism . These agents have advanced our understanding of signal transmission pathways in lymphocyte activation. Mutat Res, 1993 Oct, 297(3), 293 - 312 Genetic toxicity studies of gossypol; de Peyster A et al.; Gossypol, a polyphenolic compound extracted from cotton plants, shows promise for use as a male contraceptive, as well as a treatment for gynecological disorders, cancer, and certain microbial diseases . Before gossypol can be considered safe for widespread human use, particularly by healthy people of childbearing age, its effect on normal genetic processes should be understood . Characterization of gossypol's genotoxic potential has not been approached systematically, although numerous clinical and laboratory studies have addressed issues relating to genetic effects of gossypol . This review summarizes results of relevant studies and offers recommendations for the emphasis of future efforts to understand gossypol's genotoxicity potential . Evidence suggesting that gossypol has any genotoxic effects in mammals under normal physiologic conditions so far is weak, at best . However, several unresolved issues that are important for establishing long-term genetic safety of gossypol were uncovered by this analysis . These include the need for a better understanding of the significance of weak increases in SCE frequency seen in a number of laboratory exposure studies, and more definitive, comprehensive animal tumor bioassay data. Mutat Res, 1993 Oct, 292(2), 191 - 8 An improved estimator of spontaneous mutation rates in Luria-Delbrück fluctuation experiments; Jones ME; In estimating the spontaneous mutation rate, mu, in cultured mammalian cells, the number of mutant cells in several parallel cultures, each a clone from a single cell, is determined . Luria and Delbruck (1943), and subsequently Lea and Coulson (1949), proposed several estimators of mu using data from such experiments . These methods of analysis were originally proposed to apply to mutants arising in microbial cultures, and some of the limitations in applying them to mammalian cell cultures have been outlined by Featherstone et al . (1987), and by Kendal and Frost (1988) . For a given value of mu, the number of mutants, r, found in a culture is exceedingly variable, so that r contains very little information concerning mu . It is important, therefore, that an estimator of mu be efficient; that it not waste the limited information available in the experimental data . To this end, a new estimator is proposed which compounds the information previously utilized by the so-called P0 and median estimators . Using a spreadsheet program such as that available with MINITAB, the algorithm can be programmed in as few as 26 instructions . Tested against simulated data across a range of values of mu, the proposed compound estimator is more efficient than the currently used estimators. J Antibiot (Tokyo), 1993 Oct, 46(10), 1512 - 9 Novel microbial metabolites of the phoslactomycins family induce production of colony-stimulating factors by bone marrow stromal cells . II . Isolation, physico-chemical properties and structure determination; Kohama T et al.; Leustroducsins (LSNs) A, B and C, novel inducers of colony-stimulating factors (CSFs), were isolated from culture broth of Streptomyces platensis SANK 60191 mainly by ethyl acetate extraction and preparative reverse-phase HPLC . The molecular weights and molecular formulae of LSNs A, B and C are 641: C32H52O10NP, 669: C34H56O10NP and 669: C34H56O10NP, respectively . The structure elucidation revealed that they belong to the phoslactomycin group antibiotics, and their structures contain an alpha,beta-unsaturated delta-lactone, an amino group, a phosphate ester and a cyclohexane ring moiety . The structures differ only at the substituent bound to the cyclohexane ring. J Antibiot (Tokyo), 1993 Oct, 46(10), 1494 - 502 Screening method for colony-stimulating factor inducers using a human bone marrow stromal cell line, KM-102; Kohama T et al.; A new screening method for inducers of colony-stimulating factors (CSFs) was established using KM-102, a human bone marrow stromal cell line as the producer . In this method, the assay system which uses CSF dependent cell lines is combined with the CSF production system . Interleukin-1 (IL-1), which is known to upregulate CSF production in many cell populations, was used as a positive control for production of granulocyte CSF (G-CSF) and granulocyte-macrophage CSF (GM-CSF) . Induction in the positive controls was clearly detected within 24 hours . Activators of protein kinase C (PKC), protein phosphatase inhibitors and lipopolysaccharide (LPS) were positive in this assay system, but muramyl dipeptide (MDP) and Bestatin which are known macrophage activators, were negative . Inducers of CSFs were successfully detected using this assay method . Among 1,600 microbial strains tested, 2 actinomycete strains were found to produce active substances . One strain produces teleocidin-A, a strong activator of PKC, and the other strain produces a mixture of active compounds including three novel compounds . These three compounds do not induce terminal differentiation of HL-60 cells, suggesting that they are not teleocidin-like substances and form a new class of CSF inducers. J Mol Biol, 1993 Sep 20, 233(2), 305 - 12 Step-wise mutation of barnase to binase . A procedure for engineering increased stability of proteins and an experimental analysis of the evolution of protein stability; Serrano L et al.; We have chosen two members of the microbial RNase family, barnase and binase, which have 85% identity (17 substitutions and 1 deletion) and almost identical three-dimensional structure, to study the evolution of protein stability . The 17 residues that differ are scattered throughout the molecule . Each of the 17 differing residues has been mutated independently and the effect on protein stability analysed . Each point mutation has an effect on protein stability that ranges from +1.1 to -1.1 kcal mol-1 . These changes in energy are additive . There is no clear correlation between the type of mutation and the effect on protein stability . A multiple mutant having six of the single mutations that increase the stability of barnase is 3.3 kcal mol-1 more stable than wild type and has the same activity . There could be selective pressure to maintain proteins at a certain stability and, consequently, mutations that decrease stability tend to be counterbalanced by stabilizing mutations . Alternatively, there could simply be pressure to maintain stability above a certain level, and any further increases in stability need not be maintained during evolution . These results suggest a simple way to improve the stability of proteins: choose two homologous proteins that have high similarity, mutate individually all of the residues that differ between the two, and combine the mutations that increase the stability in a multiple mutant. Proc Natl Acad Sci U S A, 1993 Sep 15, 90(18), 8591 - 4 Cytolytic activity of intestinal intraepithelial lymphocytes in germ-free mice is strain dependent and determined by T cells expressing gamma delta T-cell antigen receptors; Kawaguchi M et al.; We have compared the cytolytic activities and the cellular compositions of the intestinal intraepithelial lymphocyte (i-IEL) populations in three different combinations of conventional (CV) and germ-free (GF) mice . Cytolytic activity of i-IELs expressing gamma delta T-cell antigen receptors (TCRs) is strain dependent in CV mice (high vs . low), and this strain-dependent variability is unaltered in the GF condition . Although absolute numbers of gamma delta i-IELs are slightly decreased, the composition of CD8 alpha alpha+ and CD4-CD8- subsets and the usage of TCR gamma- and delta-chain variable gene segments by gamma delta i-IELs remain the same in GF mice . By contrast, cytolytic activity of alpha beta TCR-expressing i-IELs is uniformly high in CV mice but attenuated sharply in the GF condition . A conspicuous decrease in the total numbers of alpha beta i-IELs is also noted, and CD8 alpha beta+ and CD4+CD8+ subsets are reduced, whereas the CD8 alpha alpha+ subset is expanded in GF mice . These results indicate that microbial deprivation preferentially influences the alpha beta i-IEL population to decrease and become noncytolytic but has little effect on the pool size or characteristics of gamma delta i-IELs . Consequently, cytolytic activity of freshly isolated i-IELs from GF mice is determined by T cells expressing gamma delta TCRs and is found to be strain dependent. Eur J Pharmacol, 1993 Sep 14, 241(2-3), 221 - 7 Radicicol, a microbial cell differentiation modulator, inhibits in vivo angiogenesis; Oikawa T et al.; Angiogenesis plays a significant role in various pathological states, including the progressive growth of solid tumors, rheumatoid arthritis, psoriasis, and diabetic retinopathy, in addition to its crucial role in embryonic development . Recent studies have revealed that an angiogenesis inhibitor is efficacious for these so-called angiogenic diseases . In the previous studies, we found that retinoids and vitamin D3 analogs, which are known to exhibit cell differentiation-modulating activity, effectively inhibit angiogenesis in vivo, thus forming the basis of our working hypothesis that a modulator of cell differentiation is capable of affecting angiogenesis . In this study, to verify this hypothesis further, radicicol (syn . monorden; 5-chloro-6-(7,8-epoxy-10-hydoxy-2-oxo-3,5-undecadienyl)-beta -resorcylic acid mu-lactone), a microbial cell differentiation modulator from a fungus, a strain of Neocosmospora tenuicristata, was examined for its anti-angiogenic activity in a bioassay system involving chorioallantoic membranes of growing chick embryos . The microbial cell differentiation modulator dose dependently inhibited embryonic angiogenesis, the ID50 value being 200 ng/egg . Radicicol also inhibited both the proliferation of and plasminogen activator production by vascular endothelial cells in the nM concentration range in a concentration-dependent manner, suggesting the possible involvement of these inhibitory effects in the anti-angiogenic action of the microbial product . These results indicate that radicicol might be a potential drug for treating different angiogenesis-dependent diseases, such as solid tumors, psoriasis, rheumatoid arthritis, and diabetic retinopathy. MMWR Morb Mortal Wkly Rep, 1993 Sep 10, 42(35), 672 - 5 Handwashing and glove use in a long-term-care facility--Maryland, 1992. {Effect of the addition of tropical fruits--pineapple (Ananas comosus) and papaya (Carica papaya)--on the production of biological silage from fish} Bello R, Cardillo E, Martinez R. Instituto de Ciencia y Tecnologia de Alimentos, Facultad de Ciencias, Universidad Central de VenezuelaMicrobial Fish Silage was produced from under-utilized fish mixed with juice and waste fruits (pineapple and papaya) at 35 degrees C . Six different products were elaborated as following: A: fish muscle with pineapple juice; B: fish muscle with fruit wastes; C: gutted fish with pineapple juice; D: gutted fish with fruit wastes; E: whole fish with pineapple juice; F: whole fish with fruit wastes . Process development was evaluated by measuring: pH, acidity, non-protein nitrogen, consistency and exudate liquid . Results indicated a slow decrease in pH value and production of acidity during 20 storage days . The addition of fruits to silage did not have any effect on these values . Silage liquefaction or hydrolysis was related to the following parameters: consistency, non-protein nitrogen and exudate liquid . The addition of fruits was related to silage liquefaction or hydrolysis, and it was measured by the consistency . Exudate liquid and non-protein nitrogen increased during storage time . However silage made from whole fish showed highest values in those parameters than other processing conditions . This results suggested that hydrolysis involve first enzymes from guts and second enzymes from muscle and head . Pineapple juice did not contribute to hydrolysis process . Mainly sensory changes in the silage occurs during first 24 hours and they were related to chemical changes . Proximal analysis did not change during silage process and microbial counts indicated the freshness of raw material used i this study. J Anim Sci, 1993 Sep, 71(9), 2310 - 21 Effect of source of supplemental nitrogen on the utilization of citrus pulp-based diets by sheep; Rihani N et al.; Two trials were conducted to examine the influence of N enrichment of dried citrus pulp (DCP) through ammoniation vs urea supplementation on digestion and feeding value of a DCP-based diet for sheep . Treatments were 1) ammoniation of DCP with urea (CP-U), 2) ammoniation with ammonium hydroxide (CP-A), 3) supplementation with urea, and 4) supplementation with horse bean (HB) . Diets were isonitrogenous (1.8% N) . In Trial 1, treatment effects on characteristics of digestion were evaluated in a 4 x 4 Latin square experiment using four rams . Source of supplemental nonprotein N (NPN) did not influence (P > .10) ruminal OM digestion . Net microbial N synthesis, N absorption, N retention, and microbial efficiency were greater (P < .05) for the CP-U than for the CP-A or urea diets . Apparent N digestibility of CP-A was 14 to 25% lower (P < .05) than that of the other diets supplemented with NPN . In Trial 2, treatment effects on lamb growth performance were evaluated in a 105-d growing-finishing trial involving 32 Sardi lambs (19 kg) . Intake was similar (P > .10) across treatments (1.13 kg of DM/d) . Daily gain of the urea-fed lambs (182 g) was higher (P < .05) than that of CP-U- or CP-A-fed lambs (138 g) . Feed efficiency and dietary NE values were similar (P > .10) among diets containing NPN . Growth performance seemed to be more directly affected by intake, rather than N economy . Enrichment of DCP through ammoniation with urea was a superior method of NPN supplementation with respect to efficiency of N utilization . However, the ultimate response to the potential benefits of such treatment may be seen only if N is truly limiting performance. Fertil Steril, 1993 Sep, 60(3), 540 - 9 Evaluation of polyacrylamide gel as substitute for human cervical mucus in the sperm penetration test; Eggert-Kruse W et al.; OBJECTIVE: To compare polyacrylamide gel as synthetic medium with human cervical mucus (CM) for the in vitro sperm-penetration test during infertility investigation . PATIENTS: One hundred sixty-nine randomly chosen couples with a median duration of infertility of 4 (range, 1 to 16) years presenting at the infertility unit of the Women's University Hospital of Heidelberg, Germany . MAIN OUTCOME MEASURES: Evaluation of sperm migration in polyacrylamide gel used in four different concentrations (1.5%, 1.6%, 1.7%, 1.8%) in the capillary tube test in parallel with CM of patients' female partners and CM of fertile donors, obtained under standardized conditions . Correlation of migration test results with outcome of semen analysis including microbial cultures and testing for local antisperm antibodies by means of the mixed antiglobulin reaction, postcoital testing, and the subsequent pregnancy rate after control for female infertility factors in a prospective study . RESULTS: Sperm ability to penetrate the synthetic medium (concerning all concentrations) correlated significantly with the penetration of human CM, although polyacrylamide proved to be a stronger barrier . Sperm velocity and duration of progressive motility were markedly reduced in polyacrylamide . Polyacrylamide results correlated with the outcome of standard sperm analyses but not with sperm antibody testing . No clear differentiation was obtained with regard to subsequent fertility (19% after 6 months), although adequate sperm migration in polyacrylamide 1.8% was significantly more frequent in the fertile group . CONCLUSIONS: In analyzing the intrinsic motility, penetration testing with polyacrylamide gel provides important information not obtained by routine sperm analysis . However, particularly with regard to immunological factors and fertility prognosis, human CM should be preferred whenever possible. Xenobiotica, 1993 Sep, 23(9), 973 - 82 Microbial models of mammalian metabolism: involvement of cytochrome P450 in the N-demethylation of N-methylcarbazole by Cunninghamella echinulata; Yang W et al.; 1 . As previously reported (Yang and Davis 1992), N-methylcarbazole (NMC) is converted to N-hydroxymethylcarbazole (NHMC), and 3-hydroxy-N-hydroxymethylcarbazole (3-OH-NHMC), two relatively stable carbinolamine metabolites by the fungus Cunninghamella echinulata (ATCC 9244) . Decomposition of these two carbinolamines yields the corresponding dealkylated metabolites, carbazole and 3-hydroxycarbazole . In the present study, the possible involvement of cytochrome P450 in the requisite N-alkyl hydroxylation reaction was examined . 2 . Carbon monoxide, a classical P450 inhibitor, markedly inhibited the formation of NHMC, as did potassium cyanide . 1-Benzylimidazole, piperonyl butoxide and SKF-525A inhibited the formation of both NHMC and 3-OH-NHMC, while beta-naphthoflavone (5,6-benzoflavone) induced their formation . 3 . The source of the oxygen atom in the metabolite NHMC was examined by GC/MS analysis of NHMC formed during incubation of NMC in H218O-enriched medium which resulted in no incorporation of labelled oxygen into the metabolite . 4 . An intermolecular isotope effect was not observed for the formation of NHMC suggesting that C-H bond cleavage is not a rate limiting step in the formation of this metabolite under the conditions examined . 5 . It was concluded that P450 enzymes may be involved in the N-demethylation of NMC catalyzed by this fungal model of mammalian metabolism, and provides further support for biochemical and mechanistic parallels between mammalian metabolism and microbial systems catalyzing phase-1 biotransformations. Br J Nutr, 1993 Sep, 70(2), 647 - 61 The effect of condensed tannins in Lotus pedunculatus on the digestion and metabolism of methionine, cystine and inorganic sulphur in sheep; McNabb WC et al.; Two experiments were conducted with sheep fed on fresh Lotus pedunculatus containing 50-55 g condensed tannin (CT)/kg dry matter . Effects of CT were assessed by comparing control sheep (CT operating) with sheep receiving a continuous intraruminal infusion of polyethylene glycol (PEG) to bind and inactivate CT . Digestion of methionine and cystine was determined using a continuous intraruminal infusion of indigestible markers, whilst plasma irreversible loss (IRL) of methionine, cystine and inorganic sulphate was determined using 35S labelling . The proportion of microbial non-NH3-N (NAN) in whole rumen digesta NAN and the IRL of reducible S from the rumen were determined using a continuous intraruminal infusion of (NH4)2(35)SO4 . The proportion of microbial NAN in whole rumen digesta NAN (0.44 v . 0.71) and the IRL of reducible S from the rumen (0.84 v . 2.49 g S/d) were lower in control than PEG sheep . PEG sheep lost 30% of ingested methionine and cystine across the rumen, whereas the control sheep lost no methionine and cystine across the rumen . Apparent absorption of methionine from the small intestine was 27% higher in control than PEG sheep, but both groups had a similar apparent absorption of cystine . The apparent digestibility of cystine in the small intestine was lower in control (0.42) than PEG (0.53) sheep, whereas the apparent digestibility of methionine was similar (0.78) for both groups . CT had no effect on plasma methionine IRL, but markedly increased the IRL of cystine (39.8 v . 22.4 mumol/min) and reduced the IRL of plasma inorganic sulphate (35.9 v . 50.2 mumol/min) . A three-pool model comparing interconversions between the three plasma metabolites showed that CT increased the flow of cystine to body synthetic reactions (36.5 v . 17.3 mumol/min) . This was due to trans-sulphuration of methionine to cystine being greater in control than in PEG sheep, whilst the oxidation of both methionine and cystine were reduced in control sheep . It was concluded that CT reduced the proteolysis of forage protein and the degradation of S amino acids to inorganic sulphide in the rumen, resulting in increased net absorption of methionine and increased utilization of cystine for body synthetic reactions in sheep with a high capacity for wool growth (and, hence, high cystine requirement). Br J Nutr, 1993 Sep, 70(2), 557 - 66 Adaptation to the digestion of non-starch polysaccharide in growing pigs fed on cereal or semi-purified basal diets; Longland AC et al.; Pigs (25-45 kg) were fed on either cereal or semi-purified basal diets supplemented with either high or low levels of sugar-beet pulp or wood cellulose (Solka-floc) . The apparent digestibility and retention of N and apparent digestibility and metabolizability of energy (GE) and the apparent digestibility of non-starch polysaccharides (NSP) and their constituent monomers were measured during weeks 2, 4 and 6 of the trial . N and GE were less well-digested, retained or metabolized from cereal basal diets than from the corresponding semi-purified diets during all three periods . NSP from sugar-beet pulp was highly digestible, unlike that from Solka-floc which was relatively poorly digested . These differences of NSP digestibility were seen more clearly when incorporated in semi-purified diets . There was no significant increase in the digestibility or retention of N, or digestibility or metabolizability of GE, or in the digestibility of sugar-beet pulp NSP with increasing time-period on the diets . In contrast, the digestibility of Solka-floc NSP tended to increase with the time-period . The digestibility of NSP from the semi-purified diet with the high level of Solka-floc inclusion was much lower than that for the low level of inclusion, indicating that microbial activity had been reduced . In conclusion, adaptation to the diets in terms of N and GE balance may be complete after 1 week, but 3-5 weeks may be necessary before stability of measurements of the digestibility of resistant NSP monomers can be obtained. Br J Nutr, 1993 Sep, 70(2), 471 - 83 An integrated, dynamic model of feed hydration and digestion, and subsequent bacterial mass accumulation in the rumen; van Milgen J et al.; Hydration of feeds and bacterial attachment to feed particles are thought to play major roles in rumen digestion of fibrous feedstuffs . The objective of the present study was to integrate these phenomena in a mechanistic model that could be used for data analysis . The proposed model was based on the conversion of biomass, where digestion end-products can be used for the synthesis of bacterial mass . Digestion of the potentially digestible fraction and subsequent accumulation of bacterial mass was based on a sequential, three-compartment model . These compartments represented substrate undergoing hydration, digestion, and bacterial mass accumulation . A fraction of the substrate was used for synthesis of bacterial mass . It was assumed that these bacteria associate either temporarily or permanently with the remaining substrate . Dacron bags containing either dry or fully-hydrated lucerne (Medicago sativa), maize (Zea mays) cobs, orchard grass (Dactylis glomerata), and wheat straw were incubated in the rumen of a steer that was infused continuously with (15NH4)2SO4 . The 15N-enrichments of isolated particle-associated bacteria and residue remaining in the bags were used to estimate bacterial attachment . Substrate remaining and microbial mass accumulation were analysed simultaneously . Hydration did not appear to limit digestion . Fractional rate of digestion and appearance of attached bacterial mass was fastest for lucerne . For lucerne, 5% of the digestion end-products were used for synthesis of bacteria that associated with the substrate, whereas for maize cobs, orchard grass, and wheat straw this was 16, 14, and 19% respectively . Less than 2% of digestion end-products were used for synthesis of bacteria that permanently remained associated with the substrate . Permanent association can occur only with the indigestible fraction, and probably represents bacterial debris . Lysis and/or detachment of bacterial cells was highest for lucerne, and was indicative of the rapid dynamics of lucerne digestion. Hum Reprod, 1993 Sep, 8(9), 1405 - 13 Determination of antisperm antibodies in serum samples by means of enzyme-linked immunosorbent assay--a procedure to be recommended during infertility investigation? Eggert-Kruse W, Huber K, Rohr G, Runnebaum B. The clinical significance of antispermatozoal antibody (ASA) testing in serum samples with an enzyme-linked immunosorbent (ELISA) technique using a commercially available kit was evaluated in 95 randomly chosen couples under infertility investigation . Results were related to many other parameters of male and female fertility, e.g . outcome of sperm analysis, testing of sperm functional capacity including the crossed in-vitro sperm-cervical mucus penetration test (SCMPT), results of a microbial screening in genital secretions, testing for local IgG- and IgA-class sperm antibodies in semen by means of the mixed antiglobulin reaction (MAR) and the subsequent pregnancy rate in a prospective study . Results of ASA testing did not show any relationship with medical history and results of clinical examination, the semen quality including sperm function tests and seminal cultures, outcome of the crossed SCMPT, and local ASA . All patients with MAR (IgG or IgA) positive ejaculates were ELISA negative in serum samples . No significant difference with regard to circulating ASA (ELISA) was found in couples with and without a subsequent pregnancy . Furthermore there was no significant difference of antibody levels in subfertile female patients, virgins (n = 36), 'normal' pregnant women (n = 39) and prostitutes (n = 40) . In summary, the results of this study clearly demonstrate that the use of this method for detection of antisperm antibodies during infertility investigation cannot be recommended. Antimicrob Agents Chemother, 1993 Sep, 37(9), 1737 - 41 Microbial conversion of avermectins by Saccharopolyspora erythraea: glycosylation at C-4' and C-4''; Schulman M et al.; Avermectins and ivermectins are glycosylated at C-4'' and C-4' by both growing and resting cells of Saccharopolyspora erythraea . The reaction is catalyzed by a glycosyltransferase which is constitutive . The enzyme uses UDP-glucose as the glycosyl donor and avermectin or ivermectin mono- and disaccharides as acceptors . Avermectin and ivermectin aglycones and erythromycin A are not substrates for the enzyme. Trends Biochem Sci, 1993 Sep, 18(9), 349 - 53 Inhibitors of Ras farnesyltransferases; Tamanoi F; Farnesyltransferase catalyses the post-translational modification of proteins by a cholesterol precursor, farnesylpyrophosphate . One of the substrates of this enzyme is the product of the ras oncogene . Recently, inhibitors of farnesyltransferase have been identified through two different approaches: microbial screens for natural compounds, and substrate analogues . These inhibitors may be useful in blocking the action of Ras proteins, in further characterizing protein prenyltransferases, and in elucidating the regulation of cholesterol metabolism. Steroids, 1993 Sep, 58(9), 396 - 9 Synthesis and 1H and 13C nuclear magnetic resonance of 16-methylene-17 alpha-hydroxypregna-1,4,9(11)-triene-3,20-dione; Garcia-Martinez C et al.; A three-step synthesis of 16-methylene-17 alpha-hydroxypregna-1,4,9(11)- triene-3,20-dione, which has been previously obtained by microbial transformation, was achieved from 3 beta-hydroxy-16 alpha, 17-epoxy-16 beta-methyl-5 alpha-pregn-9(11)-en-20-one in an overall yield of 35% . All compounds involved in the process were characterized spectroscopically and their 13C NMR shielding effects are briefly discussed. J Fla Med Assoc, 1993 Sep, 80(9), 627 - 30 Polymerase chain reaction from bench to bedside . Applications for infectious disease; Emmanuel PJ; The past decade has seen the rapid advancement of molecular biology and its application in the field of infectious diseases . The polymerase chain reaction (PCR) is a technique which allows for the in vitro amplification of DNA . The ability of this method to recognize and amplify small quantities of foreign genome from unpurified samples will revolutionize the diagnoses of microbial diseases which have previously been cumbersome or impossible . It has already been widely applied in this area, and clinical labs will soon be offering this test for selected pathogens . There are ongoing studies evaluating its use in CNS infections and for the identification of viruses in the immunocompromised host . The PCR is not without problems, particularly false positive results from contamination . Because it is so sensitive, the significance of a positive result in patients with latent or chronic infections is yet to be determined . As our knowledge and familiarity with PCR expands, it will become a powerful tool for the clinician to use to identify various infectious diseases. J Forensic Sci, 1993 Sep, 38(5), 1172 - 5 Microbial DNA challenge studies of variable number tandem repeat (VNTR) probes used for DNA profiling analysis; Webb MB et al.; DNA probes commonly used for forensic DNA profiling analysis were hybridized to Hinfl digested DNA isolated from various common microbial species . Extended exposures to light sensitive film failed to detect any DNA fragments of a microbial origin following hybridizations to radio-labeled DNA insert single-locus probes (SLPs) and nonisotopically labeled oligonucleotide SLPs. J Lipid Res, 1993 Sep, 34(9), 1593 - 602 Structure-function relationships of lipoprotein lipase: mutation analysis and mutagenesis of the loop region; Henderson HE et al.; The molecular models of two microbial lipases and human pancreatic lipase (PL) have suggested the existence of common structural motifs including a buried active site shielded by an amphipathic surface loop . In an effort to explore the role of residues comprising the loop of lipoprotein lipase (LPL), we have used site-directed mutagenesis to generate three new LPL variants . In variant LPLM1 we deleted 18 amino acids leaving a loop of only 4 residues which resulted in an LPL protein inactive against triolein substrates . In contrast, two other LPL variants with only partial deletions, involving the apical section of the loop {LPLM2 (-8 amino acids) and LPLM3 (-2 amino acids)} manifested normal lipolytic activity . These findings indicate a critical requirement for the maintenance of charge and periodicity in the proximal and distal segments of the LPL loop in normal catalytic function . This is further highlighted by the detection of a mutation in the proximal section of the loop in a patient with LPL deficiency at position 225 which results in a substitution of threonine for isoleucine . The intact catalytic activity of the partial deletion variants (LPLM2 and LPLM3) further suggests that the apical residues of the loop contribute minimally to the functional motifs of the active site . We support this postulate by showing that the conserved glycine in the apical turn section (G229) can be substituted by glutamine, lysine, proline, or threonine without significantly affecting catalytic activity. J Antibiot (Tokyo), 1993 Sep, 46(9), 1327 - 33 WB2838 {3-chloro-4-(2-amino-3-chlorophenyl)-pyrrole}: non-steroidal androgen-receptor antagonist produced by a Pseudomonas; Hori Y et al.; In the course of our search for non-steroidal androgen-receptor antagonists of microbial origin, Pseudomonas sp . No . 2838 was found to produce an inhibitor of androgen binding to its receptor . This compound, named WB2838, was isolated and identified as 3-chloro-4-(2-amino-3-chlorophenyl)-pyrrole . The IC50 value of WB2838 for partially purified rat prostate cytosol receptor was 8.0 x 10(-7) M . However, the IC50 value of WB2838 against estrogen-receptor binding was about 90-fold greater than that against androgen-receptor binding . WB2838 inhibited the growth of androgen-responsive mouse mammary carcinoma SC-3 cells in the presence of 10(-8) M testosterone at IC50 value of 4.1 x 10(-7) M . This inhibition was reversed by adding 10(-5) M testosterone to the culture medium . WB2838 also showed the inhibitory activity against the growth of the ventral prostate induced by testosterone propionate in castrated immature rats . Therefore, it was concluded that WB2838 was a non-steroidal androgen-receptor antagonist. Immunol Today, 1993 Sep, 14(9), 430 - 5 A three-tiered view of the role of IgA in mucosal defense; Mazanec MB et al.; Mucosal IgA has generally been viewed as an immune barrier to prevent the adherence and absorption of antigens . Recent studies employing polarized epithelial monolayers have suggested two additional functions for mucosal IgA . One is to neutralize intracellular microbial pathogens, such as viruses, directly within epithelial cells . The second is to bind antigens in the mucosal lamina propria and excrete them through the adjacent epithelium into the lumen, thereby ridding the body of locally formed immune complexes and decreasing their access to the systemic circulation. Klin Lab Diagn, 1993 Sep-Oct, (5), 19 - 22 {The isolation of Helicobacter pylori from the gastric juice}; Bazhenkov LG et al.; Gastric contents was examined in 49 patients with duodenal bulb ulcers . Helicobacter pylori were isolated in 83.7% of cases . H . pylori density was found to be 10(3)-10(5) microbial cells per ml . Bacterioscopic studies with gram staining helped detect H . pylori in 79.6% of cases, cultural method revealed the bacteria in 63.3%, and urease test in 4.1% of cases . Poor efficacy of urease test is explained by its poor sensitivity (10(6) microbial cells per ml or higher) . The authors analyze the frequency of H . pylori isolation in relation to the month of the examination and gastric contents coloring. FEBS Lett, 1993 Aug 30, 329(3), 336 - 40 Complete amino acid sequence of puroindoline, a new basic and cystine-rich protein with a unique tryptophan-rich domain, isolated from wheat endosperm by Triton X-114 phase partitioning; Blochet JE et al.; A new basic protein has been isolated from wheat endosperm by Triton X-114 phase partitioning . It contains five disulfide bridges and is composed of equal amounts of a polypeptide chain of 115 amino acid residues and of the same chain with a C-terminus dipeptide extension . The most striking sequence feature is the presence of a unique tryptophan-rich domain so that this protein isolated from wheat seeds has been named puroindoline . The similar phase partitioning behavior in Triton X-114 of this basic cystine-rich protein and of purothionins suggests that puroindoline may also be a membranotoxin that might play a role in the defense mechanism of plants against microbial pathogens. Proc R Soc Lond B Biol Sci, 1993 Aug 23, 253(1337), 131 - 5 Wave mechanisms of pattern formation in microbial populations; Agladze K et al.; The occurrence of spatially ordered structures plays an important role in biology (examples: morphogenesis, ecosystems, dynamics of populations, etc.) . Turing proposed a reaction-diffusion process that is the basis for most theoretical studies of stationary biological pattern formation . Now, when Turing structures are obtained in experiments (40 years after Turing's publication), it is interesting to discover whether Turing structures are the only mechanism used by nature in biological pattern formation . In microbial growth, we have found experimental evidence of an alternative to the Turing model that is based on waves displayed in excitable media . In studies of Escherichia coli populations, we observed that interacting taxis waves create motionless patterns . Taxis waves consuming two different substrates (serine and aspartic acid) were involved . Taxis waves consuming serine stop when they collide . However, those supported by consumption of aspartic were initiated at the collision line . Colliding and annihilating in turn, the waves give rise to stationary pattern formation, and wave theory provides an alternative to the classical Turing mechanism. Eur J Biochem, 1993 Aug 15, 216(1), 301 - 5 Complex of ribonuclease Sa with a cyclic nucleotide and a proposed model for the reaction intermediate; Sevcik J et al.; The structure of the complex of ribonuclease from Streptomyces aureofaciens (RNase Sa) with exo guanosine 2',3'-cyclophosphorothioate has been refined against 0.2-nm resolution synchrotron data using, as a starting model, coordinates from the RNase Sa: 2'-GMP complex . The refinement was based on all data over 1.0-0.2 nm and converged to a crystallographic R factor of 11.9% . This is the first structure of a microbial ribonuclease complexed with a 2',3'-cyclophosphorothioate, which is a thio analogue of the intermediate of the two-step reaction . However, exo guanosine 2',3'-cyclophosphorothioate is bound in a non-functional mode and is not hydrolysed . This structure therefore does not provide direct evidence on the identity of the amino acid residues responsible for catalytic cleavage of the substrate . However, based on present and previous results, a plausible model is proposed for the complex of the cyclic intermediate which acts as substrate for the second step of the catalysis. J Immunol, 1993 Aug 15, 151(4), 2273 - 82 T cell epitope expression of mycobacterial and homologous human 65-kilodalton heat shock protein peptides in short term cell lines from patients with Behçet's disease; Pervin K et al.; T cell epitopes of the 65-kDa heat shock protein (HSP) were mapped in patients with Behcet's disease (BD), by stimulating T cells with the overlapping synthetic peptides derived from the sequences of the Mycobacterium tuberculosis 65-kDa HSP . Significant lymphoproliferative responses were stimulated with four HSP peptides in BD, as compared with the related disease (recurrent oral ulcers), unrelated disease, and healthy controls (p < 0.05 to 0.005) . In order to assess the relative frequency of sensitized lymphocytes by these peptides, 7353 short term cell lines were generated from the lymphocytes of patients and controls . Peptides 111-125, 154-172, and 311-325 (p < 0.001) and peptide 219-233 (p < 0.02) yielded significantly greater frequency of STCL in BD than in healthy and disease controls . All but peptide 154-172 stimulated only the CD4+ subset of T cells, although there was no evidence that reactivity to the selected peptides is restricted by DR2 to DR7 Ag . HLA-B51 is significantly associated with BD, but there was no evidence that B51 was a restricting element, when B51+ patients were compared with B51- patients with BD, and with B51+ healthy control subjects . A comparative investigation was then carried out between the corresponding mycobacterial and human HSP peptides . Similar or higher lympho-proliferative responses were stimulated by the human peptides compared with the mycobacterial peptides . These results suggest that the four peptide determinants within the 65-kDa HSP might be involved in the pathogenesis of BD . Whereas the high microbial load and associated stress proteins found in oral ulceration of BD may initiate an immune response to these conserved epitopes, expression of autoreactive T cell clones might be stimulated by immunodominant T cell epitopes of endogenous HSP which may induce immunopathologic changes. J Chromatogr, 1993 Aug 6, 644(2), 389 - 95 Determination of ionic species formed during growth of Escherichia coli by capillary isotachophoresis; Futschik K et al.; The ionic species that are formed during the microbial growth of Escherichia coli were determined by capillary isotachophoresis as a function of the time of cultivation . This formation was indicated by the change in a sum parameter, the impedance of the nutrient broth, measured by a special electrode system . Based on the determination of the individual ions formed under the given conditions (identified as acetate, lactate, alpha-ketoglutarate, fumarate, ammonium and probably a simple amine), the change in conductivity was calculated and compared with that obtained by the impedance measurement of the bulk medium . From the results it can be concluded that the change in the sum parameter as a function of time is originated by the ions determined. J Periodontol, 1993 Aug, 64(8), 694 - 700 Comparative analysis between a modified ultrasonic tip and hand instruments on clinical parameters of periodontal disease; Copulos TA et al.; The goal of this study was to determine whether an ultrasonic scaler with a modified tip is as effective as a curet in providing supportive periodontal treatment for patients, based on clinical parameters of periodontal disease . Nine patients with 10 sites exhibiting probing pocket depth > or = 3 mm were treated at 0, 90, and 180 days in a single-blind, split-mouth design for supportive periodontal treatment with either Gracey curets (GC) or an ultrasonic scaler with a modified tip (MU) . Clinical parameters included plaque index, gingival index, bleeding on probing, darkfield microscopy, and elastase presence . Probing pocket depths and attachment levels were measured using an electronic probe . Measurements of clinical parameters were taken at 0, 14, 45, 90, 135, and 180 days . The results showed that treatment with MU was as effective as treatment with GC in all clinical parameters measured . Both treatment modalities were effective in reducing the elastase levels . Instrumentation time was significantly reduced with the MU (3.9 minutes vs . 5.9 minutes, P < 0.05) . The MU instrument effectively reduced the microbial environment in a significantly shorter time as compared to GC. J Antibiot (Tokyo), 1993 Aug, 46(8), 1219 - 31 Microbial glycosidation of some anthracycline antibiotics by an antibiotic-negative mutant of aclarubicin producer; Johdo O et al.; Microbial conversion of anthracyclinone monosaccharides using aclarubicin-negative mutant of Streptomyces galilaeus was found to produce anthracyclinone disaccharides which had either rhodinose or 2-deoxyfucose as an additional sugar . By this conversion we obtained twelve new anthracyclines from seven anthracyclines which had rhodosamine, N-monomethyldaunosamine or daunosamine at C-7 as a glycosidic sugar . All products had a reduced cytotoxic activity in comparison with those of parent compounds . However, some of them showed a therapeutically improved antitumor effects against L1210 leukemia in vivo. J Antibiot (Tokyo), 1993 Aug, 46(8), 1214 - 8 Phthoxazolin A, a specific inhibitor of cellulose biosynthesis from microbial origin . II . Isolation, physico-chemical properties, and structural elucidation; Tanaka Y et al.; Phthoxazolin A is a new inhibitor of cellulose biosynthesis produces by Streptomyces sp . OM-5714 . The active compound was isolated, and the structure was elucidated by spectrometric analyses. Inflammation, 1993 Aug, 17(4), 521 - 30 Phorbol myristate acetate induction of chemotactic migration of human polymorphonuclear neutrophils; Gabler WL et al.; The in vitro migration of human polymorphonuclear neutrophils (PMNs) was studied employing an enzymatic assay of cell migration with phorbol myristate acetate (PMA) as the test stimulant . Our data clearly show that PMA in concentrations between 1 and 100 ng/ml in the lower wells of blind-well chambers induced chemotactic migration . Chemokinesis (increased migration) was not induced when PMA was present in both the upper and lower chambers (i.e., in a nongradient mode) . Clearly our data indicate that PMA is chemotactic for human PMNs and, coupled with published studies of the effect of PMA on PMNs, suggest activation of an intracellular gradient of membrane-associated protein kinase C as a possible new mechanism for the induction of oriented migration of PMNs . Such a mechanism may be generalized to include membrane-soluble materials (e.g., inflammatory mediators, microbial products), which establish internal gradients of activated PKC rather than via the "classic" agonist-surface receptor mechanism, providing an alternative pathway for the induction of leukocyte chemotaxis. Electrophoresis, 1993 Aug, 14(8), 782 - 8 Temperature-gradient gel electrophoresis for analysis and screening of thermostable proteases; Sattler A et al.; The thermal unfolding of microbial serine proteases was studied by temperature-gradient gel electrophoresis (TGGE) . Conditions for a native polyacrylamide gel electrophoresis were established, and the temperature gradient was applied perpendicularly to the direction of electrophoretic migration . Mobility changes of the protease molecules were indicative for thermally induced conformational changes . The transition temperature was determined with good accuracy . The native and active protease conformation was detected by an activity assay in the temperature-gradient gel . As a consequence of the typical protease-autoproteolytic reaction at elevated temperatures, the unfolded protease conformation could not be detected for non-inhibited, active subtilisin . After inhibition by phenylmethylsulfonyl fluoride (PMSF) the complete structural transition could be followed by TGGE . This transition is "discontinuous", i.e . the thermal transition is either very slow, compared to the time of electrophoresis, or irreversible, as known for subtilisins from calorimetric data . Inhibition by the strong serine specific inhibitor diisopropyl fluorophosphate (DFP) led to two conformations at low temperature . One conformation is stabilized by 8 degrees C, the other by at least 20 degrees C as compared with PMSF inhibition . The influence of calcium ions on the subtilisin stability was investigated by a series of TGGE under different calcium concentrations . The strong calcium binding site is occupied even without added calcium, occupation of the weak binding site leads to a stabilization of 10 degrees C with a binding constant around 10(6) M-1 . The subtilisin Carlsberg stability could also be investigated in unpurified bacterial culture supernatants . Thus, the method is suitable for screening of thermostable subtilisin mutants directly after expression in a bacterial host.(ABSTRACT TRUNCATED AT 250 WORDS) Comput Appl Biosci, 1993 Aug, 9(4), 403 - 5 A numerical method for allocating microbial isolates to strain types when characterized by typing methods that are not 100% reproducible; Hunter PR; Many methods for typing microbial strains are not 100% reproducible . This can create problems when deciding whether different groups of isolates are really distinct or represent typing errors or variation of a single strain . Neither hierarchical clustering nor iterative partitioning methods are suited for analysing such data . A novel iterative partitioning method is described which allows for the uncertainty of the typing method in use . Before grouping strains, the maximum dimension of the groups is set based on a previous knowledge of the typing method's reproducibility . Isolates are only allocated to a group if they differ from that group's typical strain type by less than the number of reaction differences required to distinguish between two strains . In a series of Monte Carlo studies the accuracy of strain allocation was found to be very good, even when the two groups were situated close to each other. Clin Infect Dis, 1993 Aug, 17 Suppl 1, S236 - 42 Pathogenesis of lymphoid interstitial pneumonia in natural and experimental ovine lentivirus infection; DeMartini JC et al.; Ovine lentivirus (OvLV), as a member of the lentivirinae subfamily of Retroviridae, shares morphological, genomic, and cytopathic features with human immunodeficiency virus (HIV) . Although OvLV infection does not induce profound immune deficiency in sheep, it has many similarities with HIV infection, such as the capacity to infect macrophages, undergo antigenic variation in vivo, and induce slow progressive diseases involving the pulmonary, lymphoid, and central nervous systems . Studies of the pathogenesis of disease in sheep naturally or experimentally infected by OvLV are providing clues to the pathogenesis of HIV infection, including the significance of viral load, the emergence of cytopathic variants, the mechanisms and significance of viral antigenic variation, and viral neutralization, and mechanisms of lymphoproliferation and tissue destruction induced by the virus . Preliminary evidence suggests that infection by other microbial agents, including Mycoplasma species, may play a cofactor role in the pathogenesis of lentivirus-associated lymphoid interstitial pneumonia in sheep, but further studies are required to address this issue. Immunol Today, 1993 Aug, 14(8), 373 - 5 Heat shock proteins in multiple sclerosis and other autoimmune diseases; Georgopoulos C et al.; Because of the abundance of the highly conserved heat shock proteins (hsp) in microbial pathogens and in mammalian cells, hsps have been considered candidates as target antigens in autoimmune disorders such as multiple sclerosis (MS) . Consequently, this workshop examined the current understanding of the biology of hsps and discussed the evidence that they may contribute to autoimmune disease processes . This article reports the outcome of the discussion. Gastroenterol Jpn, 1993 Aug, 28(4), 541 - 6 Decreased production of active oxygen species by neutrophils in patients with liver cirrhosis and hepatocellular carcinoma; Itoh K et al.; Adult respiratory distress syndrome (ARDS) following infection is one of the postoperative complications of hepatectomy of cirrhosis . In this study we focused on the anti-microbial activity of neutrophils . We measured production of active oxygen species by neutrophils, and simultaneously examined their nutritional status, immunity and ICG (K-ICG) disappearance ratio . When compared to the controls, the patients with cirrhosis had significantly lower production of O2- upon stimulation by N-formyl-Met-Leu-Phe (fMLP) or opsonic zymosan (OZ) . The presence of cancer did not affect results for the groups studied . Overall H2O2 production was lower in cirrhotic patients than in controls . There was a positive correlation between O2- production and K-ICG, which was used to estimate the severity of cirrhosis . Indicators of nutrition and immunity were also lower in cirrhotic patients, but neither of these indicators correlated with the production of active oxygen species . From these results, we concluded that the production of active oxygen species by neutrophils is lower in cirrhotic patients than in controls . Moreover, this decline correlates with the severity of cirrhosis. Circ Shock, 1993 Aug, 40(4), 243 - 9 Prostaglandin E1 analogues misoprostol and enisoprost decrease microbial translocation and modulate the immune response; Gianotti L et al.; The aim of this study was to investigate the ability of two prostaglandin E1 (PGE1) analogues, misoprostol and enisoprost, to alter bacterial translocation following burn injury . Balb/c mice were treated with misoprostol (n = 36) or enisoprost (n = 36) for 3 days with different doses (20 or 200 micrograms/kg/day) prior to receiving a 20% full-thickness burn and simultaneous gavage with 1 x 10(10) 14C-Escherichia coli . Animals were sacrificed 4 and 24 hr postburn, and blood, peritoneal fluid, mesenteric lymph nodes, spleen, liver, and lungs were harvested aseptically . Radionuclide counts, number of viable bacteria, and percentage of translocating bacteria remaining alive in each tissue suggested that the high doses of misoprostol or enisoprost decreased the magnitude of 14C-E . coli translocation, while the low dose of both drugs enhanced bacterial clearance . Therefore, both misoprostol and enisoprost reduce bacterial translocation, and modulate bacterial clearance in a dose-dependent manner. Int J Biol Macromol, 1993 Aug, 15(4), 215 - 20 Enzymatic degradation of poly(hydroxyalkanoates) by Pseudomonas pickettii; Yamada K et al.; A bacterium capable of degrading poly(3-hydroxybutyrate) {P(3HB)} was isolated from laboratory media and identified as Pseudomonas pickettii . The strain P . pickettii excreted an extracellular poly(hydroxyalkanoate) (PHA) depolymerase and grew on P(3HB) as the sole carbon source . P . pickettii also grew on 3-hydroxybutyrate, glucose, fructose, citrate or succinate . However, only 3-hydroxybutyrate apart from P(3HB) induced the secretion of PHA depolymerase . The PHA depolymerase was purified from the culture medium containing 3-hydroxybutyrate at the sole carbon source by hydrophobic column chromatography and gel filtration, and its molecular weight was determined as about 40,000 by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate . The optimum activity of degrading P(3HB) by the depolymerase was observed at pH 5.5 and 40 degrees C . The enzymatic degradation of microbial copolyesters was studied by the weight loss (erosion) of solution-cast films of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and poly(3-hydroxybutyrate-co-4-hydroxybutyrate) . 1H-n.m.r . analysis of the water-soluble products after the hydrolysis of P(3HB) film by PHA depolymerase revealed that the main product was the monomer 3-hydroxybutyric acid. Anal Chem, 1993 Aug 1, 65(15), 2036 - 9 Detection and removal of Escherichia coli using fluorescein isothiocyanate conjugated monoclonal antibody immobilized on bacterial magnetic particles; Nakamura N et al.; A novel fluoroimmunoassay method using bacterial magnetic particles for the highly sensitive detection of bacteria has been developed . Fluorescein isothiocyanate (FITC) conjugated monoclonal anti-Escherichia coli antibody was immobilized onto bacterial magnetic particles (BMPs) using a heterobifunctional reagent, N-succinimdyl 3-(2-pyridyldithio)propionate (SPDP) . E . coli cells were reacted with FITC-antibody-BMP conjugates for 15 min in an inhomogeneous magnetic field which enhanced aggregation . The cell/BMP complexes sedimented, causing relative fluorescence intensity of the solution to decrease with increasing microbial cell concentration . A linear relationship was obtained between the relative fluorescence intensity and cell concentration in the range of 10(2)-10(6) cells/mL . Selectivity of this detection system was satisfactory . Monoclonal antibody immobilized on BMPs was also applied to the specific removal of E . coli from the bacterial suspension. Appl Environ Microbiol, 1993 Aug, 59(8), 2648 - 52 Abundance of airborne Penicillium CFU in relation to urbanization in Mexico City; Rosas I et al.; Air was sampled simultaneously at three localities in Mexico City differing in urbanization index and air pollution level on 22 days during a period covering both dry and rainy seasons . An Andersen two-stage microbial sampler was used for 15 min at 28 liters min-1 to isolate culturable fungi on malt extract agar . After exposure, plates were incubated at 25 degrees C for 48 to 72 h before colonies were counted and identified to give concentrations of total fungal spores and of Penicillium spp., expressed as CFU per cubic meter of air . Total fungi numbered 91 to 602 CFU m-3 in Tlalpan Borough (southern area), 40 to 264 CFU m-3 in Cuauhtemoc Borough (downtown), and 26 to 495 CFU m-3 in Gustavo A . Madero Borough (northern area) . Although Penicillium spp . were the second most frequently isolated fungal genus, concentrations were small, with a maximum of only 133 CFU m-3 . Twice as many colonies were isolated in the southern area, with an urbanization index of 0.25 (arithmetic mean, 41 CFU m-3), as at other sampling stations with greater urbanization indices (arithmetic means, 19 and 20 CFU m-3) . In the downtown area, with an urbanization index of 1.0, Penicillium spp . were more numerous than any other genus and formed 25% of the total fungal count compared with 14 and 17% in the other areas . Concentrations of airborne Penicillium spp . did not differ significantly between rainy and dry seasons.(ABSTRACT TRUNCATED AT 250 WORDS) J Clin Periodontol, 1993 Aug, 20(7), 528 - 36 Healing in periodontal defects treated by decalcified freeze-dried bone allografts in combination with ePTFE membranes (I) . Clinical and scanning electron microscope analysis; Guillemin MR et al.; This study clinically evaluates the use of decalcified freeze-dried bone allograft (DFDBA) in conjunction with an expanded polytetrafluoroethylene (ePTFE) membrane specifically designed for the treatment of interproximal intraosseous defects . It also examines by SEM, plaque contaminated membranes retrieved from patients . 15 advanced periodontitis patients with two bilateral interproximal probing depths of > or = 6 mm participated . After hygiene phase, measurements were made to determined soft tissue recession, pocket depth, clinical attachment levels and amount of keratinized tissue . Defects from each pair were randomly treated with ePTFE plus DFDBA (experimental) or DFDBA alone (control) . Measurements were made during the surgery to determine crestal resorption, defect resolution and defect fill . Membranes were removed at 4 to 6 weeks and analyzed by SEM . Each site was surgically reentered and measurements repeated at six months . Both groups showed clinical and statistically significant changes when compared to baseline (P < 0.01), but no difference between groups . The experimental group showed increased soft tissue recession vs control group, 0.9 versus 0.4 mm, and loss of keratinized tissue 1.6 versus 0.1 mm (P < 0.0001) . Control sites showed a 58% bone fill while experimental sites had 70% bone fill . There were no clear patterns of microbial colonization or cell adherences in either side of the membrane . It was concluded that the presence of plaque on the membranes did not compromise the initial clinical healing during the first 4-6 weeks . Results suggest a beneficial effect with the use of either technique for the treatment of intraosseous defects. J Clin Periodontol, 1993 Aug, 20(7), 471 - 5 Aging and microcirculatory dynamics in human gingiva; Matheny JL et al.; There have been no published reports characterizing gingival vascular function in humans of different ages . In this study, we compared gingival vascular dynamics in 60 healthy male humans ranging in age from 18-75 years (young, 18-25 years (Y); middle, 35-45 years (M); old, 65-75 years (O), 20 subjects/group) . Both videomicroscopy of individual microvessels and laser doppler flowmetry were employed to assess marginal gingival circulation . Systemic cardiopulmonary parameters were monitored as controls . The number of gingival vessels visible in a microscopic field was higher and the number of microvessels exhibiting active flow was lower in M and O compared to Y . There were no differences among age groups in laser doppler flow values (tissue blood flow) or in red blood cell velocity in individual vessels . Although not statistically significant with age there was a trend towards decreasing blood flow velocity in both the laser doppler and videomicroscopic measurements . Blood pressure increased slightly with age . Peripheral oxygen saturation was lower in O compared to Y and M . No differences were seen in heart rate, respiratory rate or end-tidal carbon dioxide concentration . The changes in blood pressure and oxygen saturation were expected . The differences in numbers of visible vessels and the number of vessels with active blood flow may reflect the way the gingival microcirculation adapts with age to changing nutritional needs or to microbial and/or mechanical challenge. Clin Chem, 1993 Aug, 39(8), 1608 - 12 Calcium determination in serum with stable alkaline Arsenazo III and triglyceride clearing; Morgan BR et al.; We describe an analytical procedure for determining serum calcium, using the ligand Arsenazo III in an aqueous alkaline medium . The choice of pH for the proposed technique differs from current procedures, which are for the most part carried out in a slightly acidic medium . An acidic medium avoids interference from magnesium, but is spectrophotometrically suboptimal for this pH-dependent reaction: the molar absorptivity of the Arsenazo III complex with calcium at acid pH is 13,787 L mol-1 cm-1, about one-half of that at a more optimal alkaline pH (26,574 L mol-1 cm-1) . We have included a clearing technique in the reagent to avoid spectral aberrations from hypertriglyceridemic samples . alpha-Cyclodextrin absorbs the nonesterified fatty acids liberated from triglycerides by a microbial lipase . This modification may also be helpful for binding nonesterified fatty acids, which are known to interfere with calcium procedures by forming calcium soaps and thus preventing the reaction with intended ligands . The use of 8-hydroxyquinoline sulfonate as the magnesium-masking agent facilitates the use of alkaline pH . The less-water-soluble alternative, 8-hydroxyquinoline, commonly used as a precipitating agent for several methods, is difficult to solubilize in the alkaline reagent and tends to precipitate when complexed to magnesium . Finally, the use of alkaline pH results in a prolonged (> or = 6 weeks) shelf life for the reagent. Proc Natl Acad Sci U S A, 1993 Aug 1, 90(15), 6966 - 70 Composite, large spirochetes from microbial mats: spirochete structure review; Margulis L et al.; Phenomena previously unknown in free-living spirochetes are reported: large-sized cells with variable diameter (length to 100 microns, width between 0.4 and 3.0 microns), composite structure (smaller spirochetes inside larger ones), and positive phototropic behavior . These bacteria, Spirosymplokos, are compared with all other spirochete genera . The large spirochete, grown in mixed culture, was studied live and by transmission EM . The protoplasmic cylinder was replete with spherical granules 20-32 nm in diameter, and three to six periplasmic 26-nm flagella were inserted subterminally . Comparably granulated and flagellated small spirochetes were located inside the protoplasmic cylinder and in the periplasm of the large ones . When exposed to air, movement became erratic, protoplasmic cylinders retracted to lie folded inside the outer membrane, and refractile membranous structures formed . From one to four structures per still-moving spirochete were seen . Spirosymplokos was enriched from laboratory samples exposed to oxygen-rich and desiccating, but not dry, conditions for at least 4 mo after removal of microbial mat from the field. Proc Natl Acad Sci U S A, 1993 Aug 1, 90(15), 6919 - 23 Dominant lethality by expression of a catalytically inactive class I tRNA synthetase; Schmidt E et al.; Alignment-guided mutagenesis was used to create an inactive, but toxic, aminoacyl-tRNA synthetase . An Asp-96-->Ala (D96A) replacement in the nucleotide binding fold of the class I Escherichia coli isoleucyl-tRNA synthetase inactivates the enzyme without disrupting its competence for binding isoleucine tRNA . Expression of plasmid-encoded mutant enzyme in a cell with a wild-type ileS chromosomal allele resulted in cell death . Introduction of a second K732T substitution previously shown to weaken tRNA binding gives an inactive D96A/K732T double mutant . Expression of the double mutant is not lethal to E . coli . D96A but not the double mutant significantly inhibited in vitro charging of isoleucine tRNA by the wild-type enzyme . The results suggest a dominant tRNA binding-dependent arrest of cell growth caused by a reduction in the pool of a specific tRNA . Specific tRNA binding drugs may have therapeutic applications for treatment of microbial pathogens. Am Rev Respir Dis, 1993 Aug, 148(2), 425 - 30 Inhibitory effect of staurosporine on protein kinase C stimulation of airway smooth muscle cells; Souhrada M et al.; Phospholipid/calcium-dependent protein kinase (protein kinase C {PKC}) is a critical system in signal transduction of many different cells including airway smooth muscle (ASM) cells . We have previously shown that after administration of different phorbol esters, specific activators of PKC, characteristic electrical and contractile changes of ASM cells can be demonstrated . Similarly, our data showed that stimulation of PKC is implicated in the process of sensitization and the specific antigen challenge response of ASM cells . In this study we examined the effect of staurosporine, a microbial alkaloid, which has been reported to be a specific inhibitor of PKC, on sensitization- and specific antigen challenge-induced electrical and contractile changes of ASM cells . The effect of staurosporine was compared with those of amiloride, furosemide, and compounds NA-0345 and H-7, both synthetic PKC inhibitors . We used ASM preparations isolated from adult male guinea pigs (Camm-Hartley strain) . Changes in both membrane potential (Em), measured by a glass microelectrode technique, and isometric force, measured by copper-beryllium strain gauge, were continuously monitored . Experiments were conducted with optimal length (Lmax) of ASM preparations and at 37 degrees C . We found that the exposure of ASM preparations to staurosporine, NA-0345, H-7, amiloride, and furosemide (all in 10(-12) to 10(-4) M) had no measurable effect on the resting membrane potential or isometric force of ASM preparations . In contrast, pretreatment of ASM preparations with staurosporine, NA-0345, H-7, amiloride, or furosemide significantly attenuated (p < 0.001) phorbol myristate acetate-induced changes of ASM preparations.(ABSTRACT TRUNCATED AT 250 WORDS) Am J Pathol, 1993 Aug, 143(2), 473 - 9 Villitis of unknown etiology is associated with major infiltration of fetal tissue by maternal inflammatory cells; Redline RW et al.; Villitis of unknown etiology (VUE) is a common placental lesion and an important cause of intrauterine growth retardation and recurrent reproductive failure . Two theories have been proposed to explain VUE . One proposes that VUE is an exclusively fetal immune response to microbial antigen, whereas the other suggests that maternal cells cross the maternal-fetal interface and mount an immune response to fetal antigens . To differentiate between these alternatives, we performed in situ hybridization using X and Y chromosome-specific probes and immunostaining for CD3 and CD45 on VUE placentas from male infants . A total of eight foci and 40 villi were studied from four male VUE placentas . Controls included nonaffected villi from each male VUE placenta, a female VUE placenta, and male and female tonsils . Affected villi showed a major proportion of XX (maternal) cells (range 30 to 54%) . An appropriate percentage of the remaining (fetal) cells contained Y chromosomes . The fraction of cells within the eight foci staining for CD3 (T lymphocytes) ranged from 34 to 57%, whereas the fraction staining for CD45/LCA (total leukocytes) ranged from 45 to 74% . The proportion of maternal cells within each focus was significantly correlated with the number of CD3-positive T lymphocytes but not with the number of CD3-negative leukocytes . We conclude that maternal cells, probably CD3-positive T cells, cross the maternofetal barrier and participate in VUE. Ophthalmology, 1993 Aug, 100(8), 1191 - 8 Immunocytologic findings in a case of Vogt-Koyanagi-Harada syndrome; Kahn M et al.; BACKGROUND: Vogt-Koyanagi-Harada (VKH) syndrome is a form of chronic panuveitis that often has a poor long-term visual outcome . To the authors' knowledge, there have been only two previous reports describing immunocytologic findings in the eyes of patients with VKH syndrome, all studied in relatively late stages of the disease . METHODS: The authors performed immunocytologic studies on the enucleated eyes of a patient with active VKH syndrome, using monoclonal antibodies to mononuclear subsets, major histocompatibility antigens, and viral and treponemal agents, to better understand the pathophysiologic mechanisms underlying the disease . Emulsified chorioretinal tissue also was processed for viral isolation in tissue culture and for Treponema pallidum by rabbit inoculation studies . RESULTS: The dense uveal infiltrates in active VKH syndrome are composed predominantly of T lymphocytes and HLA-DR+ macrophages . Scattered T cells and plasma cells were present in the retina . In addition, nondendritic-appearing CD1 (Leu-6) positive cells were localized in the choroid, in close proximity to choroidal melanocytes . No microbial agents were identified . CONCLUSION: Whereas the etiology of the VKH syndrome remains an enigma, the authors' immunocytologic findings are consistent with a T-cell-mediated disorder . In demonstrating CD1 (Leu-6) positive cells in the infiltrates, the authors propose a potential role for these cells in the etiology of VKH syndrome. Chest, 1993 Aug, 104(2), 552 - 9 Does the bronchoscope propagate infection? Prakash UB. The flexible and rigid bronchoscopes traverse the nasopharynx or oropharynx and carry with them the indigenous microbial flora to distal regions and may thus inoculate the tracheobronchial tree and possibly the pulmonary parenchyma . The three potential consequences of this event include: (1) onset of new infection in the tracheobronchial tree or lung parenchyma or, if the patient has preexisting infection, further spread of infection locally or to extrapulmonary sites; (2) spread of infection from one patient to another via the bronchoscope, if the methods of disinfection and sterilization are inadequate; and (3) pseudoinfection due to cross-contamination of the bronchoscope, resulting in isolation of organisms from the bronchoscopic specimens of a patient who is clinically not infected . Review of the literature indicates that the last-mentioned consequence is more commonly encountered in clinical practice . The occurrence of pseudoinfection inevitably leads to costly and time-consuming procedures to guarantee that the patients are not infected . Rigorous adherence to sterilization and disinfection procedures and a common sense approach to protecting the uninfected patients and bronchoscopy personnel from infected patients and instruments will prevent the risk of propagating infection through the bronchoscope . This can be accomplished by establishing a set of policies regarding disinfection, sterilization, and protection of uninfected patients, as well as the bronchoscopist and paramedical personnel involved in bronchoscopy. J Nat Prod, 1993 Aug, 56(8), 1431 - 3 Perfragilins A and B, cytotoxic isoquinolinequinones from the bryozoan Membranipora perfragilis; Choi YH et al.; Detailed spectroscopic and cytotoxicity data are reported for two new isoquinolinequinones, perfragilins A {1} and B {2}, which were isolated from the bryozoan Membranipora perfragilis collected in South Australia . Both compounds, which are speculated to be of microbial origin, contain a relatively uncommon thiomethyl ether functionality. Can J Microbiol, 1993 Aug, 39(8), 795 - 803 Activity of microbial peptidases, oxidases, and esterases in lake waters of varying trophic status; Alun J et al.; The range and activities of microbial enzymes present in lake water were assessed directly in cells concentrated by tangential flow filtration . A total of 108 enzymes were assayed in this study, which included tests for 60 peptidases, 20 oxidases, and 10 esterases, and 18 miscellaneous tests . In general, no trends in the range of enzymes were associated with trophic status of the lakes . However, one lake that was hypereutrophic had a greater range of enzymes than the other lakes tested . An increase in total enzyme activity (activity/mL) was recorded with an increase in trophic status of the water . The relationship between the physical and chemical attributes of each lake and microbial enzyme activities was investigated by principal component analysis . Quantitative changes between lakes in 11 of the 21 variables were shown to be closely related to changes in the enzyme activities of the lakes; total organic carbon, particulate carbon, particulate nitrogen, pH, and chlorophyll a showed the closest relationships. Trends Microbiol, 1993 Aug, 1(5), 192 - 6 Attachment factors of Bordetella pertussis: mimicry of eukaryotic cell recognition molecules; Sandros J et al.; Bacteria localize to specific target organs during infection by recognizing eukaryotic cell surface addresses . This address system exists to facilitate normal trafficking of host cells such as leukocytes . The mechanisms by which bacteria subvert host targeting codes provide a unique link between microbial pathogenesis and eukaryotic cell trafficking, and can point to a new array of antibodies and peptides of potential therapeutic value. Trends Microbiol, 1993 Aug, 1(5), 181 - 7 Microbial recognition and activation of plant defense systems; Lindsay WP et al.; Molecules released or generated during microbial entry (elicitors) are recognized by components of plant cells, ultimately resulting in the induction of a battery of plant defense responses . The molecular mechanisms underlying these signaling systems, as well as the plant defense responses they control, are becoming increasingly well characterized. Trends Biotechnol, 1993 Aug, 11(8), 353 - 9 Microbial treatment of metal pollution--a working biotechnology? Gadd GM, White C. Some of the main processes that remove, immobilize or detoxify heavy metals and radionuclides in the natural environment result from microbial activities . These activities can be harnessed to clean up toxic metal wastes before they enter the wider environment . To date, the most successful biotechnological processes utilize biosorption and bioprecipitation, but other processes such as binding by specific macromolecules may have future potential . Technologies using these processes are currently used to control pollution from diverse sources, including smelters and mine workings. Biosci Biotechnol Biochem, 1993 Aug, 57(8), 1396 - 7 Site-directed mutagenesis of chitinase from Alteromonas sp . strain O-7; Tsujibo H et al.; Chitinase (Chi85) from Alteromonas sp . strain O-7 contains the two conserved regions common to microbial and plant chitinases . We did site-directed mutagenesis of Chi85 to investigate the effects of the conserved amino acid residues on chitinase activity . We suggest that Asp-290 and Glu-292 of Chi85 may be the essential amino acid residues for the cleavage of beta-glycosidic linkage of chitin. J Biol Chem, 1993 Jul 15, 268(20), 14805 - 11 Identification of a G protein alpha subunit from Neurospora crassa that is a member of the Gi family; Turner GE et al.; Heterotrimeric G proteins, consisting of alpha, beta, and gamma subunits, are implicated in major signal transduction pathways controlling a diversity of functions in eukaryotic organisms . In the filamentous fungus Neurospora crassa, G proteins are implicated in the regulation of several environmental responses . As a first step in studying the role of G proteins in these processes, we have cloned the genes for two alpha subunits, gna-1 and gna-2, from Neurospora . The genes are located on different chromosomes and are differentially regulated during asexual development . The encoded proteins (Gna-1 and Gna-2) are the same size as members of the Gi-alpha family (approximately 40 kDa) . The Gna-1 protein sequence is 55% identical overall to members of the Gi family and contains the consensus sequences for ADP-ribosylation by pertussis toxin and incorporation of myristic acid, which are found in this group . These properties make Gna-1 the first identified microbial alpha subunit to be a member of any class . Furthermore, incubation of a N . crassa plasma membrane fraction with pertussis toxin results in ADP-ribosylation of a protein substrate which is the approximate size of Gna-1 . The predicted Gna-2 protein sequence does not share a high degree of sequence identity with the Gi class . However, the coding region contains at least one intron in a position conserved in the Gi family . We propose that the Gi family of alpha subunits is ancient and during evolution may have first appeared in filamentous fungi. J Healthc Mater Manage . 1993 Aug;11(7):14, 16, 20, passim. Thoughts on levels of microbial cleanliness and on validating disinfection procedures; Nystrom B; Efforts of the European Committee of Standardization (CEN) and the International Organization for Standardization (ISO) to harmonize production standards for medical devices have brought to the fore the need to standardize definitions and names for different levels of microbial contamination as well as validation procedures for microbial inactivation . Spaulding's categories of medical devices (critical, semi-critical and noncritical items) suggest that different levels of cleanliness can be accepted . Current terminology is imprecise and does not provide the necessary information to the user . A more appropriate one is needed . So are methods to validate microbial inactivation processes to lower levels of cleanliness than sterility . In Sweden, a couple of validation studies have been performed on bacterial inactivation in automatic washing machines which disinfect through a final hot water or steam rinse . However, more studies need to be done, especially on viral inactivation. Ann Neurol, 1993 Jul, 34(1), 8 - 17 Myoblast transfer in Duchenne muscular dystrophy; Karpati G et al.; One biceps muscle of 8 patients with Duchenne muscular dystrophy was injected at 55 sites with a total of 55 million viable, purified, and contamination-free normal myoblasts (myoblast transfer) . The other biceps of each patient was injected with a placebo to serve as a control . The procedure was blinded to the patients, parents, and investigators . Myoblasts derived from a biopsy specimen of the fathers were cultured and purified under strict conditions and carefully screened for microbial contamination . All patients received cyclophosphamide for immunosuppression for 6 or 12 months . No serious complications were observed after myoblast transfer, indicating that the procedure is safe . The overall therapeutic efficiency of myoblast transfer was poor as judged by the results in maximal voluntary force generation, dystrophin content of the muscle, magnetic resonance imaging of the muscle, and the lack of donor-derived DNA and dystrophin messenger RNA in the injected muscle . An improved efficiency of the take of myoblasts might be achieved by using younger cells and injecting the myoblasts with a myonecrotic agent (to increase the prevalence of regeneration) and a basal laminal fenestrating agent. Hum Reprod, 1993 Jul, 8(7), 1025 - 31 Antisperm antibodies in cervical mucus in an unselected subfertile population; Eggert-Kruse W et al.; To determine the incidence and the clinical significance of antisperm antibodies (ASA) in cervical secretions in an unselected subfertile population, cervical mucus samples of 192 patients with long-standing infertility were screened by means of the indirect mixed antiglobulin reaction (MAR) test allowing differentiation for immunoglobulin (Ig)A and IgG in a parallel test setting . In addition, the indirect MAR IgG test in cervical mucus was evaluated by means of IgG coated latex particles instead of sensitized erythrocytes as the indicator system . All cervical mucus samples were taken under standardized conditions . Results of ASA determination were related to microbial findings in the cervix and the outcome of sperm-mucus interaction testing in vivo and in vitro, and the subsequent fertility in a prospective study . The total incidence of cervical mucus ASA within this population was low (< 2%) . A significant correlation was found between sperm antibodies of the IgG and IgA class and of IgG ASA, determined with the standard MAR and testing with latex microspheres as indicator particles . Results were not influenced by microbial colonization of the cervix and were not markedly related to lymphocytes subset testing . All ASA positive women had a negative outcome of the post-coital test, but the majority of negative post-coital tests was not caused by local mucus antibodies of the IgG and/or IgA class . In patients with positive indirect MAR testing in cervical mucus, there was no pregnancy within an observation period of > 12 months . In conclusion, the results indicate the ASA in cervical secretions are not frequent but nevertheless are a severe cause of infertility when present.(ABSTRACT TRUNCATED AT 250 WORDS) J Antimicrob Chemother, 1993 Jul, 32 Suppl A, 69 - 75 Foreign body associated infection; Jansen B et al.; One of the major complications associated with the use of medical devices and implants is foreign body associated infection . Its significance in modern medicine and its pathogenesis are discussed . Adherence to and colonization of a foreign body material and the interference of microbial products with the host defence are critical steps leading to foreign body infections . Therapy is difficult and hence the immediate administration of antibiotics and the removal of an infected device or implant is still preferred . New strategies for the prevention of foreign body infections are briefly introduced. J Anim Sci, 1993 Jul, 71(7), 1831 - 40 Efficacy of phytase in improving the bioavailability of phosphorus in soybean meal and corn-soybean meal diets for pigs; Cromwell GL et al.; Four experiments involving 225 pigs were conducted to assess the efficacy of a microbial phytase (FINASE, Alko Ltd . Biotechnology, Rajamaki, Finland) produced by Aspergillus niger in corn-soybean meal or dextrose-cornstarch-soybean meal-based diets . In two experiments with growing-finishing pigs, fortified corn-soybean meal diets were formulated to be adequate (.50%) or inadequate (.40 or .30%) in P during the growing phase followed by adequate (.40%) or inadequate (.30%) P in the finishing phase . Mono-dicalcium phosphate was the source of supplemental P . Half the diets were supplemented with phytase (500 phytase units/g) . Rate and efficiency of gain and bone breaking strength were decreased when P-deficient diets were fed . Phytase supplementation of the low-P diets restored growth rate and feed:gain to levels that approached those of pigs fed the adequate-P control diet . Bone strength was partially restored to that of the controls . In two additional experiments, pigs were fed low-P basal diets in which all the dietary P came from soybean meal or a corn-soybean meal blend . Both diets contained .05% available P . Graded levels of monosodium phosphate were added to these diets, up to .15% added P, to establish a standard curve . Phytase was added to the basal diet at 250, 500, or 1,000 units/g . Growth rate and bone strength improved linearly (P < .01) with added monosodium phosphate and with increasing levels of supplemental phytase . Based on estimates of total and available P intakes, the highest level of phytase (1,000 units/g) increased the bioavailability of the P from 25% in the soybean meal diet to 57% in the phytase-supplemented diet, and from 15% in the corn-soybean diet to 43% in the phytase-supplemented diet . Expressed on the basis of the improvement in phytate P availability, this level of phytase converted approximately one-third of the unavailable P to an available form . The results indicate that the phytase was efficacious in improving the bioavailability of phytate P for pigs. New Microbiol, 1993 Jul, 16(3), 245 - 9 Rapid methods for microbial diagnosis of mycobacterial infections; Pirali F et al.; The Authors describe their experience in rapid diagnosis of mycobacterial infections using a combination of a radiometric blood culture (Bactec 13 A) and a nucleic acid hybridization system (Gen probe, Accuprobe) to detect and identify Mycobacteria . They found out that a high number of septicaemias in HIV positive patients are due to Mycobacterium avium, while in HIV negative subjects Mycobacterium tuberculosis is the most frequent mycobacterium. J Antibiot (Tokyo), 1993 Jul, 46(7), 1116 - 25 Isolation of trehalamine, the aglycon of trehazolin, from microbial broths and characterization of trehazolin related compounds; Ando O et al.; Trehalamine, (3aR,4R,5S,6S,6aS)-2-amino-4-(hydroxymethyl)-3a,5,6,6a- tetrahydro-4H-cyclo-pent{d}oxazole-4,5,6-triol (1) and D-glucose were obtained by acid hydrolysis of trehazolin (3), a trehalase inhibitor produced by actinomycetes . More vigorous hydrolytic treatment of trehazolin afforded an aminocyclitol, (1R,2S,3R,4S,5R)-5-amino-1- (hydroxymethyl)cyclopentane-1,2,3,4-tetraol (2) . Trehalamine, the aglycon of trehazolin, was also found in the culture broths of two trehazolin producing strains, Micromonospora sp . SANK 62390 and Amycolatopsis sp . SANK 60791 . These trehazolin related compounds trehalamine (1) and 2 were poor inhibitors of trehalase (1; IC50 1.8 x 10(-4) M, 2; > 5.0 x 10(-4) M) . On the other hand they inhibited more potently rat intestinal sucrase (1; IC50 6.8 x 10(-5) M) and sweet almond beta-glucosidase (2; IC50 5.6 x 10(-6) M) than trehazolin. Appl Environ Microbiol, 1993 Jul, 59(7), 2304 - 10 Effect of treated-sewage contamination upon bacterial energy charge, adenine nucleotides, and DNA content in a sandy aquifer on Cape Cod; Metge DW et al.; Changes in adenylate energy charge (ECA) and in total adenine nucleotides (A(T) and DNA content (both normalized to the abundance of free-living, groundwater bacteria) in response to carbon loading were determined for a laboratory-grown culture and for a contaminated aquifer . The latter study involved a 3-km-long transect through a contaminant plume resulting from continued on-land discharge of secondary sewage to a shallow, sandy aquifer on Cape Cod, Mass . With the exception of the most contaminated groundwater immediately downgradient from the contaminant source, DNA and adenylate levels correlated strongly with bacterial abundance and decreased exponentially with increasing distance downgradient . ECAS (0.53 to 0.60) and the ratios of ATP to DNA (0.001 to 0.003) were consistently low, suggesting that the unattached bacteria in this groundwater study are metabolically stressed, despite any eutrophication that might have occurred . Elevated ECAS (up to 0.74) were observed in glucose-amended groundwater, confirming that the metabolic state of this microbial community could be altered . In general, per-bacterium DNA and ATP contents were approximately twofold higher in the plume than in surrounding groundwater, although ECA and per-bacterium levels of A(T) differed little in the plume and the surrounding uncontaminated groundwater . However, per-bacterium levels of DNA and A(T) varied six- and threefold, respectively, during a 6-h period of decreasing growth rate for an unidentified pseudomonad isolated from contaminated groundwater and grown in batch culture.(ABSTRACT TRUNCATED AT 250 WORDS) J Clin Microbiol, 1993 Jul, 31(7), 1876 - 81 Curvilinear-gradient high-performance liquid chromatography for identification of mycobacteria; Guthertz LS et al.; Over a 1-year period, 502 mycobacterial cultures submitted to the Microbial Diseases Laboratory were identified by high-performance liquid chromatography (HPLC) in parallel with standard biochemical methods . Identification by HPLC using a curvilinear gradient was achieved by comparing the chromatograms of the unknown cultures to chromatograms for known reference strains, together with calculation of peak height or peak area ratios, as necessary . The overall agreement between HPLC and biochemical identification was 97.2% . In addition, 7 of 12 cultures of Mycobacterium bovis were identified by HPLC as the BCG strain . Of 111 cultures biochemically identified as members of the M . avium complex (MAC), 108 were confirmed as MAC by DNA probe and 106 were confirmed by HPLC . Of the latter 106, 58 probe-positive strains were identified as M . avium, 38 were identified as M . intracellulare, and 10 were identified as Mycobacterium sp . strain "X" by HPLC . Of the remaining five nonchromogenic cultures, four had MAC-like chromatograms that did not match any in our library sufficiently to permit definitive identification . Of the latter four, two were confirmed as MAC strains by DNA probe and two were not . The last of the cultures biochemically identified as MAC (1 of 111) was a mixture of MAC and non-MAC strains . Overall, only 2 of 502 cultures yielded results by HPLC that differed from those obtained by standard biochemical methods . The HPLC result was confirmed in both cases by an independent national reference laboratory . In the 12 instances in which HPLC did not provide identification, the chromatograms were either uninterpretable or did not match available reference chromatograms . These findings show that the identification obtained by HPLC concurs well with that obtained by both the standard biochemical methods and the DNA probes . Thus, identification by HPLC provides mycobacteriology laboratories with a reproducible and specific method for accurate and timely identification of most medically important mycobacteria. Dent Clin North Am, 1993 Jul, 37(3), 457 - 63 Restorative dentistry . Interactions with periodontics; Page L et al.; The integration of periodontal considerations with restorative planning is now the standard of care . Instilling motivation toward prevention continues to be one of our most challenging goals . A stable healthy periodontium and minimum restorative needs are the resulting rewards . With the use of available knowledge and microbial testing for both periodontitis and caries susceptibility, we now can offer our patients a higher likelihood of success in retaining natural teeth and implants. AORN J, 1993 Jul, 58(1), 84 - 8 Effects of nail polish on microbial growth of fingernails . Dispelling sacred cows; Baumgardner CA et al.; Nail polish worn on short, healthy nails does not appear to be associated with increased microbial counts on the fingernails . Additional studies to examine the effect of wearing nail polish on other aspects of hand hygiene may be warranted, however. Med Trop (Mars), 1993 Jul-Sep, 53(3), 391 - 2 {Prevalence of human immunodeficiency virus infection (HIV) in patients with microbial abscesses}; Strecker W et al.; From 1987 1/1 to 1989 6/30, 80 patients with purulent abscesses underwent serological test . Seropositivity rate of HIV was 20% i.e . higher of a 2.4 factor than seropositivity rate of 8.2% found in the local population not showing any symptoms of AIDSPIP: Purulent infections are the most common pathology in tropical surgery, and drainage of pus is the most frequent surgical intervention in the Third World . Between June 1987 and June 1989, 3003 operations were performed at the General Referral Hospital of Gbadolite in northern Zaire . 637 of the operations were for infections, including 302 laparotomies for tubo-ovarian abscess and 335 drainages of microbial abscesses of various types . Serologic testing for HIV infection was conducted on 34 women with tubo-ovarian abscesses, of whom 7 (20.6%) were seropositive, and on 42 men and 4 women with microbial abscesses, among whom 9 (19.6%) were seropositive . The seroprevalence rate in the local population represented by 370 blood donors and 297 patients without clinical symptoms of AIDS was 8.2% . The seroprevalence rate among the patients with abscesses was 2.4 times higher than that of the general population . The factors favoring purulent infections in seropositive persons are not known . Nat Immun, 1993 Jul-Oct, 12(4-5), 235 - 49 Regulation of natural killer function by nonlymphoid cells; Shau H et al.; Natural killer (NK) cells and interleukin-2 activated NK cells, termed lymphokine-activated killer (LAK) cells, can lyse tumor cells without restriction by the MHC complex . The cytotoxic activity of these cells is subject to regulation by nonlymphoid cells in vitro . Interaction of NK cells with red blood cells (RBC) increases NK cytotoxicity . Interaction with polymorphonuclear neutrophils (PMN) suppresses LAK development and also inhibits NK- and LAK-mediated cytotoxicity at the effector cell phase . Exposure to tumor necrosis factor-alpha greatly enhances the inhibitory effect of PMN at the effector cell phase . Depending on their states of activation and differentiation, monocytes have the capacity to either enhance or suppress LAK induction . In contrast, macrophages derived from the lung are potent inhibitors of NK and LAK activity at both the induction and effector cell phases . Several cytokines are secreted by monocytes and macrophages and these include both suppressive and enhansive factors of cytotoxic function . Platelets are also capable of releasing suppressive factors . NK cells are believed to be involved in host surveillance of tumors, control of microbial infections and regulation of hematopoiesis . Regulation of NK cells by nonlymphoid cells and their products likely provides an in vivo mechanism for locally regulating NK function. Biosci Biotechnol Biochem, 1993 Jul, 57(7), 1231 - 3 Stimulation of nerve growth factor production by pyrroloquinoline quinone and its derivatives in vitro and in vivo; Yamaguchi K et al.; Pyrroloquinoline quinone (PQQ), which is a cofactor of microbial quinoprotein enzymes, was found to be a potent enhancer of nerve growth factor (NGF) production in vitro . One of PQQ derivatives, oxazopyrroloquinoline trimethylester, had little activity in vitro, but increased the NGF content in rat brain in vivo. J Physiol, 1993 Jul, 466, 329 - 44 Proton currents in human granulocytes: regulation by membrane potential and intracellular pH; Demaurex N et al.; 1 . To determine whether conductive pathways contribute to the H+ efflux from granulocytes, we used the whole-cell patch-clamp technique combined with microfluorimetric determinations of cytosolic pH (pHi) in single, dimethylsulphoxide-differentiated HL-60 cells . 2 . In voltage-clamp mode, depolarization of the cell from the resting potential (around -60 mV) to +60 mV caused an increase in pHi that was accompanied by a sizeable outward current . 3 . Ion substitution experiments and analysis of the reversal potential of tail currents indicated that the outward current is carried largely by H+ ions . 4 . Full activation of the H+ current occurred within 1-2s after depolarization and deactivation within 100-200 ms upon repolarization . 5 . This H+ conductance was strongly dependent on pHi, being larger at acidic pH . In addition, at low pHi the threshold for voltage activation of the H+ conductance was shifted to more negative values . 6 . Addition of millimolar concentrations of Cd2+ and Zn2+ to the bath solution reduced the maximum H+ conductance and shifted the voltage dependence of the H+ conductance to more positive potentials . The effects were reversible . 7 . In conclusion, our results demonstrate that granulocytic HL-60 cells possess a voltage-gated and pHi-sensitive H+ conductance . Because both a depolarization and a cytosolic acidification occur during the activation of granulocytes, this conductance may play a role in pHi homeostasis of granulocytes during microbial killing. Mutat Res, 1993 Jul, 297(1), 61 - 95 A critical review of the genotoxic potential of electric and magnetic fields; McCann J et al.; 55 published articles were identified which reported results of tests of ELF (extremely low frequency) or static electric or magnetic fields for genotoxic effects . The biological assays used spanned a wide range, including microbial systems, plants, Drosophila, mammalian and human cells in vitro and in vivo . Experimental results were grouped into four exposure categories: ELF Electric; ELF Magnetic; Static Electric; and Static Magnetic . The internal electric fields present in media (for in vitro experiments) and in the torso and extremities (for in vivo experiments) were estimated, providing an index of comparison . All experiments were critically analyzed with respect to basic data quality criteria . Experiments within each exposure category were then compared to determine if results reinforced or contradicted one another . The preponderance of evidence suggests that neither ELF nor static electric or magnetic fields have a clearly demonstrated potential to cause genotoxic effects . However, there may be genotoxic activity from exposure under conditions where phenomena auxiliary to an electric field, such as spark discharges, electrical shocks, or corona can occur . In addition, two unconfirmed reports suggest the genotoxic potential of certain chemical mutagens or ionizing radiation may be affected by co-exposure to electric or magnetic fields . Certain exposure categories are not represented or are under-represented by tests in some genotoxicity test systems that are usually included in minimal test batteries as specified by EPA for chemicals . It is suggested that consideration be given to whether additional genotoxicity testing is warranted to fill these gaps. Ann N Y Acad Sci, 1993 Jun 23, 685, 330 - 5 Molecular mechanisms of immunosuppression by cyclosporins; Zenke G et al.; Despite the successful clinical application of the immunosuppressive drug cyclosporin A (CsA, Sandimmun), its precise mechanism of action in the process of T cell activation remains elusive . CsA binds to the high-affinity cytosolic receptor cyclophilin whose peptidyl-prolyl cis-trans isomerase activity is inhibited upon binding . The linkage of this effect with the inhibition of the T cell receptor-mediated signal transduction pathway, which leads to a suppression of lymphokine gene transcription, is still unclear . We analyzed the relationship between cyclophilin-binding and immunosuppressive activity (e.g., effect on IL-2 transcription) of cyclosporin derivatives in vitro . The results show that binding to cyclophilin is required, but not sufficient for immunosuppression . Cyclosporin analogues which completely lack immunosuppressive activity but fully retained their cyclophilin-binding capacity antagonize the immunosuppressive activity of CsA . These derivatives inhibit the isomerase activity of cyclophilin, which clearly demonstrates that inhibition of the cyclophilin isomerase activity does not lead to immunosuppression . In analogy to the other immunosuppressants of microbial origin, FK-506 and rapamycin, a specific structure of the "effector" domain of CsA, which is unrelated to the cyclophilin-binding domain, determines the biological activity . In the nucleus, CsA interferes with the DNA-binding of inducible transcription factors to their respective DNA motifs within lymphokine promoters by affecting intracellular translocation of transcription factor subunits. J Theor Biol, 1993 Jun 21, 162(4), 447 - 63 An unexpected correlation between cardinal temperatures of microbial growth highlighted by a new model; Rosso L et al.; A new model for the prediction of microbial-specific growth rate as a function of temperature is presented . The four parameters of this model are the three cardinal temperatures (Tmax, Tmin and Topt) and the specific growth rate at the optimum temperature (mu opt) . A comparison with three other models was made on the basis of several criteria (simplicity and biological significance of parameters, applicability, quality of fit, minimum structural correlations and ease of determination of parameters) . A detailed comparison of a 217-point data set, and an extensive comparison of 47 different data sets show that the new model is better than its competitors . The three cardinal temperatures were found to be independent of mu opt . A very strong and unexpected linear correlation between the cardinal temperatures was observed . The consequences of this biological result are discussed, even though causes remain unknown. Transfusion, 1993 Jun, 33(6), 458 - 65 Identification of the blood component responsible for increased susceptibility to gut-derived infection; Gianotti L et al.; It has previously been reported that the transfusion of allogeneic whole blood increases sepsis-related mortality and decreases the ability of the host to kill bacteria that have translocated from the intestinal tract . To determine which blood component contributes to this adverse effect, the impact of the transfusion of white cells (WBCs), red cells (RBCs), and plasma on microbial translocation, bacteria killing, and mortality rate was studied . Blood from C3H/HeJ mice was separated into WBCs, RBCs, and plasma, and these fractions were transfused to Balb/c mice . Controls received sterile saline . Five days after transfusion, all Balb/c mice underwent a 20-percent burn and gavage with 1 x 10(10) Escherichia coli labeled with 14C-glucose . Mortality was observed for 10 days . Four additiona