Microbiological

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 additional groups, receiving the same treatment as above, were sacrificed 4 hours after the burn, and mesenteric lymph nodes, liver, kidney, and blood were harvested aseptically . For each tissue, quantitative colony counts, radionuclide counts, and percentage of translocated bacteria that remained alive were calculated . By radionuclide counts, no difference was observed in the degree of 14C E . coli translocation among the groups . In contrast, the percentage of viable bacteria and the mortality rate were significantly higher in the group receiving allogeneic WBCs than in all other groups (p < 0.05) . It is concluded that WBCs are the component in transfused blood that has an adverse effect on host resistance to gut-derived infection.

Infect Immun, 1993 Jun, 61(6), 2681 - 6
A bacterial protease perturbs the paracellular barrier function of transporting epithelial monolayers in culture; Azghani AO et al.; Tight junctions between cells and adhesion to the substratum maintain the barrier function of epithelia throughout the body . Damage to the epithelial barrier by microbial products allows penetration of bacteria and promotion of infection . We studied the effects of Pseudomonas elastase (PE) on the barrier function of epithelia by using Madin-Darby canine kidney (MDCK) epithelial cells; these cells form tight junctions (zonula occludens {ZO}) in vitro . PE decreased electrical resistance across the monolayers in a concentration- and time-dependent manner . Immunostaining of selected proteins of the ZO and zonula adherens was used to explore the effects of PE on junctional proteins . PE-treated monolayers of MDCK cells had markedly decreased immunostaining of ZO-1, a protein of the ZO, but light microscopy of PE-treated cells revealed no obvious morphologic changes . A chromium release assay indicated that, even with marked changes in transmonolayer electrical resistance, the permeability defect was not due to membrane disruption . Fluorescence staining of F-actin indicated diminution of cellular microfilaments in PE-treated cells, but E cadherin (uvomorulin), a protein of the zonula adherens, was unaffected by the enzyme . Elastases from porcine pancreas and human leukocytes with similar enzymatic activity (6 U/ml) did not decrease transmonolayer electrical resistance or degrade ZO-1 . These results suggest that PE disturbs the barrier function of epithelial monolayers, in part, by changing the cell architecture and altering at least one protein of the ZO.

Fiziol Zh Im I M Sechenova, 1993 Jun, 79(6), 115 - 27
{Disordered permeability of the gastrointestinal tract barrier for macromolecules and the possibilities for its experimental dietetic correction}; Gmoshinskii IV et al.; Macromolecular absorption of food and microbial antigens being enhanced in the intestine under pathological conditions may well be the cause of such diseases as food allergy, coeliac disease, Crohn's disease, atopic eczema, etc . The polyethyleneglycol-4000 (PEG-4000) and food antigens absorption was found to be similar in the animal intestine . The PEG-4000 gastrointestinal permeability is considerably increased in the rats with anaphylaxis, experimental biliary malabsorption and experimental colitis . Fatty acids of the omega-3 and omega-6 series as well as histidine were found to change the permeability for the PEG-4000.

J Periodontol, 1993 Jun, 64(6), 553 - 8
Short-term sequential administration of amoxicillin/clavulanate potassium and doxycycline in the treatment of recurrent/progressive periodontitis; Matisko MW et al.; The systemic use of a single antibiotic was compared to that of a sequential antibiotic regimen in the treatment of A . actinomycetemcomitans and/or P . gingivalis-associated periodontitis . Eleven patients with recurrent/progressive periodontitis and demonstrating subgingival infection with A.a . and/or P.g . were selected . Six patients received oral administration of doxycycline (Do), 200 mg the first day and 100 mg for 4 days thereafter, and then amoxicillin/clavulanate potassium (Au), 500 mg 3 times daily for 5 days . The other 5 patients received only doxycycline for 10 days . Eight sites with > or = 5 mm probe depth per patient were selected, of which 4 received root planing at time 0 . Clinical measurements (GI and PI, probing pocket depth, probing attachment level, and bleeding upon probing/suppuration) and microbial infection levels (2 sites/patient as per DNA probe) for A.a . and P.g . were recorded at 0, 4, 12, and 25 weeks . Clinical data were subjected to statistical analysis of variance and t-tests for significance . The Do + Au groups produced significant reduction in probing pocket depth (PPD) at 4, 12, and 25 weeks (1.1, 1.3, and 1.1 mm, respectively) . The Do group produced significant reduction in PPD only at 4 and 12 weeks (0.8 and 0.8 mm); the Do + Au group produced significant gain of 0.8 mm in probing attachment level at 4 and 12 weeks; and the Do + Au group in conjunction with root planing produced the most sustained reduction in PPD and gain in PAL . These findings suggest that the sequential use of multiple antibiotic agents may offer greater promise as an adjunctive treatment approach for the management of recurrent and/or progressive periodontitis than a single antibiotic regimen.

J Anim Sci, 1993 Jun, 71(6), 1657 - 65
Influence of level of urea and method of supplementation on characteristics of digestion of high-fiber diets by sheep; Rihani N et al.; Effects of level and method of urea supplementation of N utilization and characteristics of digestion of a N-deficient (.85% N), citrus pulp-based diet were evaluated in a 4 x 4 Latin square design experiment, using four ruminally and intestinally cannulated crossbred (D'man x Sardi) rams . Two levels of urea (15.1 and 8.8 g/d, providing total N intakes of 12.0 and 9.1 g/d, respectively) and two methods of supplementation (urea mixed with the diet or continuously infused in the rumen) were compared in a 2 x 2 factorial arrangement of treatments . Ruminal pH varied between 6.2 and 7.0 across treatments and tended (P > .10) to be lower in animals receiving urea infusion . Ruminal ammonia concentrations were lower (P < .01) and more stable with urea infusion than with urea feeding . Ruminal VFA concentrations and profiles were unaffected (P > .10) by treatments . Site and extent of OM and fiber digestion were not influenced (P > .10) by either level of urea in the diet or method of supplementation . Average ruminal OM and ADF digestion were 53.1 and 64.5%, respectively . Total tract digestion values averaged 75.9 and 73.5%, respectively . With the exception of N efficiency (duodenal nonammonia N/total N intake) that was 31% lower (P < .10) for the high- than for the low-urea treatment, there were no treatment responses or interactions (P > .10) on characteristics of N utilization . Microbial N flow to the small intestine averaged (across treatments) 9.0 g/d and constituted 87% of the nonammonia N leaving the abomasum.(ABSTRACT TRUNCATED AT 250 WORDS)

J Nutr, 1993 Jun, 123(6), 1117 - 23
Supplemental microbial phytase improves bioavailability of dietary zinc to weanling pigs; Lei X et al.; Two experiments were conducted to determine the effects of supplemental microbial phytase on utilization of dietary zinc by weanling pigs . Experiment 1 was a 2 x 3 factorial arrangement of treatments with 24 pigs for 4 wk . Two levels of phytase activity (0 and 1350 units/g) and three levels of zinc (0, 30 and 60 mg/kg as ZnSO4.7H2O) were added to a corn-soybean meal basal diet . Weekly measures included growth performance, plasma alkaline phosphatase activity and plasma mineral concentrations . In Experiment 2, mineral balances were determined in 12 pigs fed the basal diet or the diet with added zinc (30 mg/kg) or phytase (1350 units/g) . The results indicated that either supplemental phytase or supplemental zinc increased plasma alkaline phosphatase activity and plasma zinc concentrations, but these increases were not additive . Supplemental phytase decreased plasma alkaline phosphatase activity in pigs supplemented with zinc . Supplemental phytase also significantly enhanced weight gain, feed intake, gain:feed ratio, plasma concentrations of inorganic phosphorus, and retention of phosphorus and calcium . Neither supplemental phytase nor zinc affected zinc retention . Supplementing corn-soybean meal diets with microbial phytase at 1350 units/g feed improves bioavailability of zinc as well as of phytate phosphorus to weanling pigs.

Med Hypotheses, 1993 Jun, 40(6), 377 - 8
Prevention of AIDS in HIV-infected people by means of T cells stimulated with microbial superantigens?
Elomaa E.
It has been shown that neuroleukin, a secretary product of T cells, is partly homologous to HIV gp 120 envelope protein . Since HIV naive T cells sometimes induce the production of antibodies against HIV coat proteins in a new host, it is suggested that the development of AIDS in HIV-infected people could be prevented by means of T cells stimulated with superantigens of either bacterial or viral origin.

Rinsho Byori, 1993 Jun, 41(6), 719 - 25
{Studies on the fragments of FDP in 3 patients with DIC associated with acute promyelocytic leukemia}; Okumura N et al.; We previously reported a study on fibrinolysis and fibrinogenolysis that analyzed fragments of fibrin/fibrinogen degradation products (FDP) using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting . In this report, we characterized the fragments of FDP in three patients with disseminated intravascular coagulation (DIC) associating with acute promyelocytic leukemia (APL) . D, Y, DD, DY/X and high molecular weight fragments were observed in sera of all the patients obtained at the onset of APL . These results showed that various degrees of fibrinogenolysis, concomitant with fibrinolysis, was occurring in APL patients presenting DIC . However, changes in the patterns of the FDP fragments during anticoagulation therapy were apparently different among three patients . Namely, fibrinogenolysis was dominant in case 1, while fibrinolysis was dominant in case 2 . Interestingly, fibrinogenolysis and fibrinolysis were almost equivalent from the onset to the end of DIC in case 3 . In case 3, FDP and FDP-D dimer were remarkably elevated about two months before the onset of APL, although their elevation was not complicated with DIC but with bone marrow necrosis . At that time, serum LDH levels and plasma polymorphonuclear elastase (PMN-Ela) were increased presumably due to the release of these enzymes from necrotic bone marrow, and the levels of CRP and plasma fibrinogen were increased probably due to an infectious complication . In non-DIC period of case 3, FDP and FDP-D dimer were spontaneously decreased without reduction of PMN-Ela levels, after three weeks of chemotherapy for microbial agents . Taken together, characteristics of FDP fragments were unique to each case of APL-DIC, probably because many factors differently affected the degradation of fibrin and/or fibrinogen.

Immunol Invest, 1993 Jun, 22(4), 267 - 82
Induction of experimental autoimmune myasthenia gravis with acetylcholine receptors using a nonionic block copolymer as adjuvant; Shenoy M et al.; To induce autoimmune diseases in animals, the auto-antigen has to be emulsified in adjuvants (e.g., complete Freund's adjuvant) containing microbial products such as Mycobacterium tuberculosis . But these powerful immunoadjuvants are not without undesirable immune response to the microbial proteins and induction of adjuvant arthritis, which could interfere with the antigen specific autoimmune response to be tested . This study was performed to evaluate the requirement of microbial products in the induction of experimental autoimmune diseases, and to identify an adjuvant without unwanted immune responses . C57BL/6 mice were inoculated with Torpedo acetylcholine receptors (T-AChR) emulsified in Titermax (TM), an adjuvant containing nonionic block copolymer and no microbial products, and evaluated for experimental autoimmune myasthenia gravis (EAMG) susceptibility . Mice immunized with T-AChR in TM demonstrated characteristic myasthenic muscle weakness with electrophysiological defect, elevated serum anti-AChR antibodies, and muscle AChR loss . None of the mice that received TM alone had muscle weakness, serum anti-AChR antibodies or muscle AChR loss . The data imply that microbial products are not critical in the induction of autoimmune diseases like myasthenia gravis in mice . Further, nonionic block copolymer could be an ideal adjuvant in the induction of autoimmune diseases in animals.

Zentralbl Bakteriol, 1993 Jun, 278(4), 522 - 8
Optimal selection of biochemical tests to identify microbial species; Claus H; Microbial species are mainly identified by biochemical tests . A numerical procedure is described, serving as an aid to decide which biochemical tests should be used and to check if these tests are sufficient to discriminate the species under consideration.

Vet Parasitol, 1993 Jun, 48(1-4), 261 - 71
The potential for avermectins to affect the nutrient economy of grazed pastures; King KL; This examination of the potential ecotoxic effects of the avermectins in temperature pastures grazed by sheep is based on a community approach and is focussed on one important aspect of ecosystem function, the nutrient cycle . Data on the amount and distribution of sheep dung on pastures grazed at different stocking rates indicated that areas of high stocking and sheep camps would be affected by avermectin residues to the greatest extent . Mineral losses from sheep dung which does not contain ivermectin, have been examined to provide a background against which the potential effects of avermectins on nutrient cycling can be appraised . The source of the diet is important; for example, dung from sheep grazing on improved pasture loses sulphur faster, and has higher microbial activity, than that of native pasture, regardless of whether it is fresh or old . Dung-dwelling fauna such as dung beetles and microarthropods are most abundant in areas of high dung concentration and microbial activity is greatest in sheep camps where a large quantity of excreta is voided . However, while there is evidence that avermectin does affect certain of the larger dung-dwelling fauna, little is known of its effects on the smaller invertebrate biota such as free-living nematodes and microarthropods . Calculations for phosphorus budgets on both native and improved pastures indicate that the amount of phosphorus recycled in these systems could be reduced by up to 5% the dosage levels currently recommended for the drug.

Int J Biol Macromol, 1993 Jun, 15(3), 189 - 92
Recognition of microbial cells by the surface of polymers with oxyethylene units; Zhaoxin L et al.; The recognition of microbial cells by the surface of polymers with oxyethylene units was investigated using the organism Trichoderma reesei, in which the surface was modified by the radiation polymerization method . The cells were recognized by their adhesion to the surface of the polymer giving a bioaffinity similar to cell fusion . The adhesion was affected by the molecular structure of the surface relating to the length of the oxyethylene unit, and enzyme productivity from the adhered cell walls was affected by the property of the polymer.

J Anim Sci, 1993 Jun, 71(6), 1648 - 56
Effect of bird-resistant and non-bird-resistant sorghum grain on amino acid digestion by beef heifers; Streeter MN et al.; The effects of sorghum type on amino acid digestion were determined by feeding heifers (230 kg) equipped with ruminal, duodenal, and ileal cannulas bird-resistant (BR) or non-BR grains that had a normal or waxy endosperm (NORMAL-BR, WAXY-BR, NORMAL, WAXY) . Dry-rolled grain diets were fed at 2% of BW in a 4 x 4 Latin square design . Total (TAA), essential (EAA), and nonessential (NAA) amino acid intake (grams/day) tended to be greater for BR than for non-BR grains . Feed (plus endogenous) amino acids reaching the duodenum were calculated by subtracting amino acids of microbial protein from total flow . Flow (grams/day) of total and feed TAA, EAA, NAA, and proline-rich-protein (sum of aspartate, glutamate, glycine, and proline) to the duodenum was greater (P < .05) for BR than for non-BR grains . The ratio between feed proline-rich-protein and NAA reaching the duodenum was similar to that noted with feed intake . Amino acid disappearance (grams/day) from the small intestine was unchanged (P > .10) by sorghum type, although the digestibility (percentage of duodenal flow) of TAA, EAA, NAA, and most individual amino acids was decreased (P < .05) for BR varieties . Amino acid flow to the cecum (grams/day) was generally greater for NORMAL-BR than for WAXY-BR (P < .10) or for NORMAL (P < .01) and greater (P < .10) for WAXY-BR than for WAXY.(ABSTRACT TRUNCATED AT 250 WORDS)

Anal Biochem, 1993 Jun, 211(2), 288 - 92
Matrix-assisted ultraviolet laser desorption/ionization mass spectrometry applied to multiple forms of lipases; Hedrich HC et al.; Matrix-assisted ultraviolet laser desorption/ionization mass spectrometry was used to investigate heterogeneous patterns and molecular masses of microbial lipases from Penicillium camembertii, Geotrichum candidum, and Pseudomonas sp . Mass spectral peaks of the native, glycosylated lipases from P . camembertii and G . candidum were broader than those of the corresponding deglycosylated enzymes, indicative of heterogeneous glycosylations . The broader peaks in the mass spectra were caused by an overlapping of unresolved peaks, derived from single glycoprotein species . Molecular masses determined for the deglycosylated proteins were in excellent agreement with those deduced from amino acid composition and sequence data, whereas with conventional biochemical methods (gelfiltration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis) only very rough estimations of molecular masses were possible . By mass spectrometric analysis of the four fractions of chromatographically separated P . camembertii lipase molecular masses of 29,990, 34,030, 31,990, and 32,140 Da were found before and 29,960, 29,980, 29,990 and 30,010 Da, respectively, after deglycosylation . Thus from the four native fractions of P . camembertii lipase three were glycoproteins . G . candidum lipase showed an average molecular mass of 63,500 Da for the heterogeneously deglycosylated native form and a molecular mass of 59,650 Da for the deglycosylated enzyme . For the Pseudomonas lipase, which could only be isolated with lipids firmly attached, a molecular mass of 32,890 Da was determined, in close agreement with that derived from the cDNA sequence.

Am J Obstet Gynecol, 1993 Jun, 168(6 Pt 1), 1654 - 64; discussion 1664-8
The relationship between spontaneous rupture of membranes, labor, and microbial invasion of the amniotic cavity and amniotic fluid concentrations of prostaglandins and thromboxane B2 in term pregnancy; Romero R et al.; OBJECTIVE: The purpose of this study was to examine the relationship between rupture of membranes, labor, and microbial invasion of the amniotic cavity and amniotic fluid concentrations of eicosanoids in patients with spontaneous rupture of membranes at term . STUDY DESIGN: Amniotic fluid was retrieved by transabdominal amniocentesis from patients with rupture of membranes and patients with intact membranes at term . Studies to determine the microbial state of the amniotic cavity included culture for bacteria and mycoplasmas, Gram stain, amniotic fluid white blood cell count, and Limulus amebocyte lysate . Eicosanoids (prostaglandin E2, prostaglandin F2 alpha and its stable metabolite, 6-keto-prostaglandin F1 alpha, and thromboxane B2) were determined with sensitive and specific radioimmunoassays validated for human amniotic fluid . Statistical inference was conducted with analysis of variance and linear contrast . RESULTS: (1) Spontaneous rupture of membranes at term was associated with a significant increase in amniotic fluid concentrations of all eicosanoids measured in this study except 6-keto-prostaglandin F1 alpha . (2) Early labor in patients with rupture of membranes was associated with a significant increase in the amniotic fluid concentration of all eicosanoids . (3) A significant increase in amniotic fluid eicosanoids in women with microbial invasion of the amniotic cavity could not be documented . CONCLUSIONS: Whereas preterm labor in the absence of microbial invasion of the amniotic cavity is not associated with a significant increase in amniotic fluid concentrations of prostaglandins, a clear increase was documented in women with early labor after spontaneous rupture of membranes . These observations suggest that there are fundamental differences in the biochemistry of term and preterm parturition.

Clin Oral Implants Res, 1993 Jun, 4(2), 53 - 64
Microbial colonization on natural tooth structure compared with smooth and plasma-sprayed dental implant surfaces; Gatewood RR et al.; The purpose of this study was to characterize and compare by scanning electron microscopy (SEM) the maturation of supragingival microbial plaque on enamel and smooth titanium and that of subgingival plaque on cementum, plasma-sprayed titanium and hydroxyapatite surfaces . Specimens (6.5 x 2 x 1 mm) were obtained from titanium implants with smooth transmucosal collars and plasma-sprayed endosseous posts of titanium or hydroxyapatite . Unerupted third molars were sectioned to similar dimensions that included equal lengths of enamel and root surface . Ten patients with post-treatment pocket depths of > 6 mm on 3 non-adjacent teeth were selected and each had 1 set of the 3 specimen types bonded by random assignment to the selected teeth . Specimens were positioned so that enamel and/or smooth titanium surfaces were supragingival and cementum and titanium or hydroxyapatite plasma-sprayed surfaces were subgingival . Patients were instructed to stop oral hygiene, after which 2 specimen sets were removed at 1, 3, 5, 7 and 10 days . Two specimens of each type were examined by SEM at each time interval for supra- and subgingival plaque . The sequence of appearance of various microbial morphotypes in supra- and subgingival plaque was similar regardless of surface . In both supra- and subgingival plaque, depending on time interval, cocci, rods of various lengths, filamentous organisms, fusiforms, spirochetes and corn-cob formations were observed . Mineralized plaque was noted on most subgingival specimens by day 10 . These observations indicate that surface of natural teeth, smooth titanium and plasma-sprayed titanium and hydroxyapatite support the maturation of microbial plaque with the successive inclusion of a variety of morphotypes.

J Exp Med, 1993 Jun 1, 177(6), 1779 - 84
Promoter of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon gamma and bacterial lipopolysaccharide; Xie QW et al.; Inducible nitric oxide synthase (iNOS) can be expressed by many types of mammalian cells in response to diverse signals acting synergistically, including cytokines and microbial products . We previously showed that induction of iNOS in mouse macrophages by interferon gamma (IFN-gamma) and lipopolysaccharide (LPS) was at the transcriptional level . From a mouse genomic library, we now cloned a 1,749-bp fragment from the 5'-flanking region of the iNOS gene, and used S1 nuclease mapping and primer extension to identify the mRNA transcription start site within it . The mRNA initiation site is preceded by a TATA box and at least 22 oligonucleotide elements homologous to consensus sequences for the binding of transcription factors involved in the inducibility of other genes by cytokines or bacterial products . These include 10 copies of IFN-gamma response element; 3 copies of gamma-activated site; 2 copies each of nuclear factor-kappa B, IFN-alpha-stimulated response element, activating protein 1, and tumor necrosis factor response element; and one X box . Plasmids in which all or the downstream one half or one third of this region of iNOS were linked to a reporter gene encoding chloramphenicol acetyltransferase were transfected into cells of the RAW264.7 macrophage-like line . All these constructs conferred inducibility of the iNOS promoter by LPS, but only the construct containing all 1,749 bp conferred synergistic inducibility by IFN-gamma plus LPS.

J Mol Biol, 1993 May 20, 231(2), 161 - 6
Phenotypic suppression by incorporation of an alien amino acid; Lemeignan B et al.; Azaleucine is a naturally occurring amino acid antibiotic that can be incorporated into proteins by mimicking leucine . In spite of its highly toxic character, the contrast between the hydrophobic side-chain of leucine and the basic side-chain of azaleucine suggested a mechanism for rescuing certain mutants . We constructed a thymidine auxotrophic mutant of Escherichia coli by replacing an arginine residue with leucine in the catalytic centre of thymidylate synthase, and indeed showed that activity could be restored by incorporation of azaleucine . This result extends the current scope of phenotypic suppression to mischarging with amino acid analogues . Microbial strains with a clear-cut requirement for an additional amino acid, as reported here, should be instrumental for widening the genetic code experimentally.

Tidsskr Nor Laegeforen, 1993 May 20, 113(13), 1566 - 8
{Frequency and clinical consequences of self-injection among salmon vaccinators}; Dyrkorn R et al.; As general practitioners in one of Norways largest fish farming regions, we carried out a prospective study of 12 vaccinators of salmon in order to determine the frequency of self-injection, and the subsequent clinical reactions . The study was carried out in close cooperation with the local responsible veterinarian . We recorded 2,430,615 vaccinations of vaccine in salmon and smolt during the spring of 1992, with 12 accidental self-injections among the vaccinators . Clinical reactions were reported in four cases . There were only moderate local reactions . In our opinion, most of the reactions were of an inflammatory nature and were not caused by microbial infection . We question the first-choice, routine treatment with antibiotics that has been practised by physicians.

Eur J Pharmacol, 1993 May 19, 236(2), 263 - 8
The neuroprotective properties of ES-242s, novel NMDA receptor antagonists, in neuronal cell culture toxicity studies; Nozawa M et al.; ES-242-1, a novel bioxanthracene of microbial origin, blocked glutamate-induced neuronal death in a dose-dependent manner at concentrations ranging from 0.01 to 1.0 microM, but not the neuronal death caused by kainic acid or quisqualic acid . ES-242-1 also prevented cell death induced by 2,4-methanoglutamate, which is a specific agonist for the NMDA receptor . ES-242-1 showed protective effects in cultured neurons prepared from cerebellum and septum as it did in cultured hippocampal neurons but to different extents . There was a positive correlation between the potencies of ES-242s as inhibitors of ligand binding to the NMDA receptor and as inhibitors of neuronal death . Hypoxic treatment for 4 h under 95% N2 and 5% CO2 caused neuronal death of the cultured hippocampal neurons . Again, ES-242-1 at 1.0 microM was effective to protect neurons against hypoxic injury . ES-242 compounds are new chemical entities possessing neuroprotective properties useful in the treatment of diseases involving glutamate toxicity.

J Dairy Sci, 1993 May, 76(5), 1353 - 64
Ruminal, intestinal, and total digestibilities of nutrients in cows fed diets high in fat and undegradable protein; Palmquist DL et al.; To study relationships of high undegradable intake protein and dietary fat on intestinal AA supply, the ruminal, intestinal, and total digestibilities of diets with or without added fat (5% of DM) and animal protein (blood meal: hydrolyzed feather meal, 1:1; 8% of DM) were examined with four cows in a 2 x 2 factorial design in a 4 x 4 Latin square experiment . Ruminal degradabilities were 14.9 and 18.6%, and intestinal digestibilities were 98.9 and 68.3%, respectively, for CP in blood meal and feather meal . Treatment effects on ruminal digestibilities were small . Protein supplementation increased total N intake by 29%, duodenal AA N flow by 39%, and AA N absorbed by 37%; absorption of Leu and Lys increased 60 and 33%, and absorption of Ile and Met increased 11 and 7%, respectively . Measured duodenal AA N flow (Cr2O3 marker) was 33% higher in cows cannulated adjacent to the pylorus compared with cows cannulated 100-cm distal to the pylorus, but only when cows were fed protein-supplemented diets; the estimates from those diets caused calculated microbial protein efficiency to exceed theoretical values . We postulated that blood meal and feather meal segregated near the pylorus, yielding high estimates of duodenal AA N flow . Removal of data for protein-supplemented diets obtained from cows cannulated at the pylorus yielded estimates of microbial protein synthetic efficiency consistent with literature values . Microbial synthesis of AA N was related linearly to ruminal digestion of carbohydrate . Location of intestinal cannulas may influence accuracy of nutrient flow estimates.

J Anim Sci, 1993 May, 71(5), 1281 - 7
Ruminal distribution of zinc in steers fed a polysaccharide-zinc complex or zinc oxide; Kennedy DW et al.; Six Holstein steers (initial BW 335 kg) were used in a replicated 3 x 3 Latin square design to study the distribution of Zn in ruminal contents from a conventional Zn source (ZnO) or Zn sequestered to a polysaccharide (PC-Zn) . Treatments consisted of a basal diet (no Zn supplementation) and the basal diet supplemented with Zn from either PC-Zn or ZnO . Steers were provided with 208, 920, or 896 mg of Zn/d from basal, PC-Zn, or ZnO, respectively . On collection day, steers were dosed with 800 mg of Zn from each Zn supplement and whole ruminal contents (WRC) were sampled at 1, 2, 4, 6, and 8 h postdosing . The cell-free (soluble) Zn concentrations in steers fed the PC-Zn or ZnO supplements decreased (P < .05) rapidly after dosing . Mean Zn concentrations of the cell-free fraction were considerably lower than either microbial or particle fractions of WRC . Compared with ZnO, the PC-Zn source resulted in higher (P < .05) cell-free Zn at 1 and 2 h after dosing and higher (P < .05) Zn concentrations (micrograms/gram of DM) in the fluid- and particle-associated microbial and particle fractions . Compared with basal, fluid-associated microbial DM concentration tended to be higher (P < .10) at 6 and 8 h with PC-Zn and ZnO . Results indicate a rapid disappearance of Zn from the soluble fraction and a high concentration of Zn in the DM of the microbial and particle fractions of WRC . Furthermore, PC-Zn resulted in higher levels of Zn in all fractions of WRC, and Zn supplementation tended to increase microbial DM production at later sampling times.

J Clin Microbiol, 1993 May, 31(5), 1213 - 6
Comparative study of procedures for isolation and cultivation of Legionella pneumophila from tap water in hospitals; Reinthaler FF et al.; For the isolation and cultivation of Legionella pneumophila from tap water in hospitals, we compared different media and selection techniques . A second part of the study compared the L . pneumophila yields from different water samples at identical sites . A total of 210 water samples (500 ml each) were collected from two selected sites in each of 21 hospitals . Warm water samples were collected after flow times of 0, 5, 10, and 15 min; in addition, one cold water sample was collected . Filtration was used to concentrate all samples . Following filtration, 0.1 and 1 ml each of untreated samples, heat-treated samples (3 min, 59 degrees C), and acid-treated samples (pH 2.2, 15 min) were spread onto the selective media MWY (SR 118; Oxoid) and BMPA alpha (SR 111; Oxoid), and samples from 12 hospitals were also spread onto GVPC medium (SR 152; Oxoid) . A total of 72 (34%) of the 210 samples from 12 hospitals were positive . With respect to the positive Legionella cultures, there was no significant difference between the selective media MWY, BMPA alpha, and GVPC . With the BMPA alpha supplement, more samples were positive following heat treatment (P < 0.05) or acid treatment (P < 0.05) than without any further treatment . For the maximum yield of Legionella colonies with minimum additional microbial flora, acid treatment was the most effective, and by all methods, the GVPC supplement was the most selective . For routine water tests in hospitals for differentiating between systemic and local contamination, acid treatment of the concentrated samples, the use of different selective media, and the correct selection of sampling sites are recommended.

Transplantation, 1993 May, 55(5), 1144 - 50
Characterization of human anti-porcine "natural antibodies" recovered from ex vivo perfused hearts--predominance of IgM and IgG2; Ross JR et al.; Hyperacute rejection is a major obstacle to successful transplantation of vascularized xenogeneic organs and is believed to be mediated at least in part by performed xenoreactive "natural antibodies" (NAb) . In this study, human NAb that could be involved in hyperacute rejection of pig heart xenografts were identified and characterized using an ex vivo model in which pig hearts were perfused with whole blood from individual human or pig donors . This ex vivo perfusion model allows for the continuous monitoring of physiologic parameters of cardiac function as well as sequential sampling of tissue and blood . Pig hearts perfused with allogeneic pig blood maintained normal function for at least 4 hr, whereas those perfused with xenogeneic AB+ human blood never achieved normal function and rejected completely after 30 min . In three separate experiments involving different human blood donors and pig hearts, sequential samples of perfused blood revealed a progressive depletion of anti-porcine NAb . Samples of all three rejected cardiac xenografts were homogenized, and the specifically bound human anti-porcine antibodies were eluted with citric acid . The eluted antibodies were enriched approximately 50-120-fold for anti-porcine reactivity compared with serum from the corresponding donor . Eluates contained NAb of predominantly IgM and IgG2 isotypes . Immunofluorescence histology confirmed the deposition of IgM and IgG2 but not other IgG subclasses in the rejected pig hearts . Since IgG2 utilized predominantly in response to bacterial polysaccharide antigens, our findings are consistent with the possibility that some NAb arise via crossreactivity with microbial antigens and are predominantly directed against carbohydrate rather than protein antigens.

Ophthalmology, 1993 May, 100(5), 666 - 8
Chronic dacryocystitis caused by Mycobacterium fortuitum; Artenstein AW et al.; PURPOSE: To characterize the clinical presentation of Mycobacterium fortuitum dacryocystitis, an unusual microbial cause of this disease . METHODS: The authors present a detailed description of a case of M . fortuitum chronic dacryocystitis and a review of the literature . FINDINGS: Although M . fortuitum is a well-recognized cause of chronic keratitis and corneal ulcer, it has only been reported once previously as a cause of dacryocystitis . Multiple factors that alter the ocular environment of the host may predispose to infection with this organism . The diagnosis requires isolation of the organism in the appropriate clinical setting . The authors' case represents the first patient successfully treated with dacryocystectomy . CONCLUSION: M . fortuitum is a rare cause of chronic dacryocystitis that may respond favorably to surgical therapy alone.

J Bacteriol, 1993 May, 175(10), 2943 - 51
A bifunctional enzyme, with separate xylanase and beta(1,3-1,4)-glucanase domains, encoded by the xynD gene of Ruminococcus flavefaciens; Flint HJ et al.; Adjacent regions of a Ruminococcus flavefaciens 17 DNA fragment were found to encode xylanase and beta(1,3-1,4)-glucanase activities . Sequencing of this fragment showed that both activities are encoded by a single 2,406-bp open reading frame corresponding to the xynD gene . The predicted product has a characteristic signal sequence that is followed by an amino-terminal domain related to family G xylanases, while the carboxyterminal domain is related to beta(1,3-1,4)-glucanases from several other bacterial species . These two domains are connected by a region of unknown function that consists of 309 amino acids and includes a 30-amino-acid threonine-rich sequence . A polypeptide having a molecular weight of approximately 90,000 and exhibiting xylanase and beta(1,3-1,4)-glucanase activities was detected in Escherichia coli cells carrying the cloned xynD gene . This is one of the first cases in which a microbial polysaccharidase has been shown to carry separate catalytic domains active against different plant cell wall polysaccharides within the same polypeptide . xynD is one of a family of related genes in R . flavefaciens that encode enzymes having multiple catalytic domains, and the amino terminus of XYLD exhibits a high degree of similarity with the corresponding regions of another xylanase, XYLA, which carries two different xylanase catalytic domains.

Br J Clin Pharmacol, 1993 May, 35(5), 485 - 90
Neutrophils and mononuclear cells from patients with chronic granulomatous disease release nitric oxide; Condino-Neto A et al.; 1 . Chronic granulomatous disease (CGD) is a group of genetic disorders characterised by recurrent severe suppurative infections due to impaired microbial killing . The principal biochemical defect is an impairment in the production of reactive oxygen intermediates by phagocytes . 2 . Nitric oxide (NO) is synthesised from the guanidino nitrogen atom(s) of L-arginine and has recently been proposed to be involved in defence mechanisms . The aim of this study was to investigate the involvement of the oxidative burst in the biosynthesis of NO by neutrophils and mononuclear cells from patients with CGD . 3 . NO synthesis was assayed by the ability of neutrophils and mononuclear cells to inhibit thrombin-induced washed platelet aggregation while superoxide anion (O2-) production was measured spectrophotometrically by the superoxide dismutase inhibitable reduction of cytochrome c . 4 . Neutrophils and mononuclear cells from patients with CGD released NO . This release was inhibited by nitro-L-arginine methyl ester but could be reversed by L-arginine . Zymosan- and PMA-induced O2- production was less than 10% as compared with healthy controls . 5 . These results indicate that O2- production is not essential for NO synthesis in human leucocytes.

J Dairy Sci, 1993 May, 76(5), 1324 - 33
Interactions of fiber and nonstructural carbohydrates on lactation and ruminal function; Feng P et al.; Four Holstein cows averaging 147 DIM and fitted with ruminal and duodenal cannulas were used in a 4 x 4 Latin square trial to determine diet effects on DMI, milk production, and ruminal metabolism . Diets contained either rapidly or slowly degraded NDF, referred to as low fill and high fill, respectively, combined with two percentages of nonstructural carbohydrate . Treatments were 39% nonstructural carbohydrate (low or high fill) and 29% nonstructural carbohydrate (low or high fill) . Intake of DM was not affected by either fill or nonstructural carbohydrate . Ruminal NDF digestibilities averaged 43.1 and 35.6% for the low fill and high fill diets, respectively . Ruminally digested nonstructural and total carbohydrate increased, but milk production decreased, as nonstructural carbohydrate increased from 29 to 39% in diets . Liquid and solid ruminal passage rates, as measured by Co-EDTA and Yb, respectively, were reduced by either 39% nonstructural carbohydrate or low fill diets . Lower microbial N flow to the duodenum and lower efficiency of microbial growth also were observed for diets with 39% nonstructural carbohydrate . The combination of 39% nonstructural carbohydrate and rapidly degraded fiber gave the highest DM and nonstructural carbohydrate digestion in the rumen but resulted in low microbial N synthesis per day and the least microbial N per kilogram of OM digested.

Peptides, 1993 May-Jun, 14(3), 629 - 32
Microbial cell-wall contaminants in peptides: a potential source of physiological artifacts; Majde JA; Microbial cell-wall products (MCWP) such as endotoxins are easily introduced into peptides produced under standard laboratory conditions . Because these products stimulate the induction of cytokines and other mediators, which, in turn, trigger a broad range of physiological responses . MCWP in peptide preparations are potential sources of artifacts . This brief tutorial outlines the physical/chemical nature of MCWP, some of their sources, their physiological effects, and a simple method to control for them in some peptide preparations.

Jpn J Cancer Res, 1993 May, 84(5), 574 - 81
ED-110, a novel indolocarbazole, prevents the growth of experimental tumors in mice; Arakawa H et al.; A new indolocarbazole compound, ED-110, which was obtained by glucosylating a microbial product (BE-13793C) and is a potent topoisomerase I inhibitor, showed characteristic inhibitory effects on the growth of 12 human tumor cell lines tested . The IC50 values of ED-110 against 9 of the 12 lines ranged from 11.5 micrograms/ml to 0.07 microgram/ml, while the remaining 3 lines were quite resistant (IC50, > 100 micrograms/ml) . In in vivo experiments, i.p . treatment with ED-110 increased the survival period by more than two-fold in mice implanted i.p . with P388, L1210, L5178Y or EL4 murine leukemic cells . The minimum effective dose increasing the life-span of mice bearing P388 leukemia by 25% was < 2.5 mg/kg/day x 10 and the maximum tolerated dose was > 160 mg/kg/day x 10 . ED-110 was also effective against the spontaneous metastasis of mouse Meth A fibrosarcoma cells and the growth of xenografted MKN-45 human stomach cancer cells as well as s.c . implanted mouse colon 26 and IMC carcinoma cells . These results indicated that ED-110 may have potential as a new antineoplastic agent with a large chemotherapeutic index and a wide range of effective doses.

J Periodontol, 1993 May, 64(5 Suppl), 474 - 84
Role of matrix metalloproteinases in human periodontal diseases; Birkedal-Hansen H; Matrix metalloproteinases (MMP) are a family of proteolytic enzymes that mediate the degradation of extracellular matrix macromolecules, including interstitial and basement membrane collagens, fibronectin, laminin, and proteoglycan core protein . The enzymes are secreted or released in latent form and become activated in the pericellular environment by disruption of a Zn(++)-cysteine bond which blocks the reactivity of the active site . The major cell types in inflamed and healthy periodontal tissues (fibroblasts, keratinocytes, endothelial cells, and macrophages) are capable of responding to growth factors and cytokines, as well as to products released from the microbial flora by induction of transcription of 1 or more MMP genes . Cytokines that are likely to regulate expression of MMP genes in periodontal tissues include IL-1, TNF-alpha, and TGF-alpha . In addition, triggered PMN leukocytes which express only 2 MMP (PMN-CL and Mr 92K GL) release these enzymes from specific granule storage sites in response to a number of stimuli . The evidence that MMP are involved in tissue destruction in human periodontal diseases is still indirect and circumstantial . Cells isolated from normal and inflamed gingiva are capable of expressing a wide complement of MMP in culture and several MMP can be detected in cells of human gingiva in vivo . In addition, PMN-CL and Mr 92K GL are readily detected in gingival crevicular fluid from gingivitis and periodontitis patients . Osteoclastic bone resorption does not appear to directly involve MMP, but a body of evidence suggests that bone resorption is initiated by removal of the osteoid layer by osteoblasts by means of a collagenase-dependent process.

J Periodontol, 1993 May, 64(5 Suppl), 461 - 6
Apoptosis in human monocytes: possible role in chronic inflammatory diseases; Mangan DF et al.; Apoptosis or programmed cell death is a physiological form of cell suicide that is profoundly influenced by the extracellular microenvironment . Apoptosis is characterized by a cascade of genetic and biochemical events that cause cell shrinkage, condensation of cytoplasmic and nuclear material, cleavage of chromosomal DNA into oligonucleosomesized (approximately 200 bp) fragments, and enhanced recognition of the dying cell by phagocytes . Apoptosis differs fundamentally from necrosis, the pathological form of cell death, in which the cell swells and lyses . The capacity to selectively induce apoptosis in leukocytes might be an important physiological mechanism for controlling accumulation of these cells in inflammatory lesions . In this paper, we review our data on apoptosis in human monocytes, cells which contribute both to the persistence and resolution of chronic inflammation . Apoptosis can be initiated when monocytes are cultured in the absence of appropriate exogenous stimulation . For example, addition of chemotactic factors is insufficient to block apoptosis . However, apoptosis in monocytes can be inhibited by adherence in the presence of serum, by microbial products such as lipopolysaccharide, or by certain pro-inflammatory cytokines, such as interleukin 1 and tumor necrosis factor-alpha . Cytokines derived from type 1 helper T cells (e.g., interferon-gamma) inhibit apoptosis whereas those derived from type 2 helper T cells (e.g., interleukin-4) enhance apoptosis in activated monocytes . Thus, cytokines derived from monocytes as well as T cells modulate apoptosis, implicating both autocrine and paracrine regulatory circuits in monocyte survival . The capacity to therapeutically regulate monocyte apoptosis promises to have tremendous value in promoting rapid healing or reducing the immunopathogenesis of chronic inflammation.

New Horiz, 1993 May, 1(2), 271 - 8
Pathogenesis, prevention, and management of catheter-associated infections; Bjornson HS; Infection of vascular catheters is one of the leading causes of nosocomial bacteremia in the critically ill patient . Most catheter-associated infections result from exogenous microbial contamination of the catheter at the time of insertion or during use and are endemic . Prevention of catheter-associated infection is based on measures designed to eliminate the potential for microbial contamination of the skin at the catheter insertion site, the catheter hub, tubing connectors, and any inline devices that may be present . Development and implementation of catheter-care protocols for use in the ICU should be effective in preventing catheter infections . The diagnosis of catheter infections is difficult, as there are few signs or symptoms that are specific for an infected catheter . Catheter infection should be suspected in patients who develop fever, chills, and leukocytosis with no other apparent site of infection.

Biochem Biophys Res Commun, 1993 Apr 30, 192(2), 747 - 53
Aspergillus ficuum phytase: complete primary structure elucidation by chemical sequencing; Ullah AH et al.; The primary structure of Aspergillus ficuum phytase was deduced from overlaps in peptide sequences . The unglycosylated enzyme is a 441 residue protein with a molecular mass of 48.5-KDa, as calculated from the total covalent structure . The estimated pl of the protein is about 4.76 . Of the 19 Asn residues, 9 were found to be glycosylated . The phytase consists of 37% non-polar, 42% polar, 11.5% acidic, and 9.5% basic amino acids . The putative active site of the enzyme containing the sequence RHG is located at the N-terminal region of the molecule and shows homology to the active site of both microbial and mammalian acid phosphatases, and phosphoglycerate mutase.

Cell Immunol, 1993 Apr 15, 148(1), 198 - 207
Oligopeptides of three to five residues derived from uveitopathogenic sites of retinal S-antigen induce experimental autoimmune uveitis (EAU) in Lewis rats; Sunil S et al.; We have previously reported that microbial peptides having 3 to 6 amino acid sequence homology with uveitopathogenic peptide of retinal S-antigen (S-Ag) are capable of inducing experimental autoimmune uveitis (EAU) in Lewis rats and subhuman primates . T-cells obtained from animals immunized with these microbial peptides proliferate when stimulated with S-Ag peptide in culture and vice versa . We were interested to know whether these 3 to 6 identical amino acid residues by themselves were sufficient to induce EAU in susceptible animals under optimal conditions . For this purpose we have determined the response of synthetic oligopeptides of 3 to 18 amino acid residues for their ability to indice EAU . Here, we report that small synthetic oligopeptides of 3 to 5 amino acid residues of the two uveitopathogenic peptides of S-Ag induced EAU in Lewis rats, although with a higher dose (2000 micrograms/rat) than that of the larger peptides (50 micrograms/rat) . These results define the core sequences in the uveitopathogenic peptides of retinal S-Ag.

J Gen Microbiol, 1993 Apr, 139 ( Pt 4), 889 - 95
Comparison of the fatty acid profiles of Borrelia, Serpulina and Leptospira species; Livesley MA et al.; Fatty acid methyl ester (FAME) derivatives were examined as a means of characterizing Borrelia burgdorferi isolates and distinguishing them from other spirochaetes . Analysis was performed using a gas liquid chromatography column in conjunction with Microbial Identification System (MIS) software . Reproducible FAME profiles were produced which distinguished Borrelia species, Serpulina hyodysenteriae and Leptospira icterohaemorrhagiae . Furthermore, the FAME profiles of four recognized Borrelia species (including two American isolates of Borrelia burgdorferi, B31 and JD1) were distinct from one another and from the BSK II medium in which they were grown . The results confirm previous reports that FAME profiles of bacteria represent a diagnostic phenotypic property and suggest that they may have applications in the chemotaxonomic classification of Borrelia species.

Vet Immunol Immunopathol, 1993 Apr, 36(3), 265 - 79
Modulation of the intracellular survival of Brucella abortus by tuftsin and muramyl dipeptide; Price RE et al.; Tuftsin, a physiologic bioactive peptide of animal origin, and muramyl dipeptide, a synthetic bioactive glycopeptide of microbial origin, are known to enhance several recognized macrophage functions and increase non-specific resistance of the host against a number of pathogens . The influence of these two bioactive peptides was studied in permissive bovine mammary macrophages that were unable to control the intracellular replication of Brucella abortus and restrictive bovine mammary macrophages that were able to effectively reduce the intracellular survival of B . abortus . Addition of tuftsin (Thr-Lys-Pro-Arg) or muramyl dipeptide significantly (P < 0.03) enhanced the ability of the permissive macrophages to control the intracellular replication of B . abortus strain 2308 and resulted in the functional conversion of the permissive macrophages into restrictive macrophages . Addition of tripeptide tuftsin fragment (Lys-Pro-Arg), a natural inhibitor of tuftsin, to the medium completely abrogated the effect of tuftsin (P < 0.03) . No additive effect on the ability of the macrophages to control the survival of B . abortus resulted from the combination of tuftsin and muramyl dipeptide.

Mikrobiyol Bul, 1993 Apr, 27(2), 164 - 70
{Superantigens and antigen recognition of T lymphocytes}; Gulmezoglu E; Superantigens are antigens which can stimulate T cells bound to MHC molecules . The conventional foreign antigens are recognized by the T cell within the MHC peptide binding groove . Superantigens differ from conventional antigens . They bind with high affinity to class II MHC molecules outside the antigen binding groove in the absence of antigen processing . The MHC class II/superantigen complexes on antigen presenting cells trigger the proliferation of T cells expressing the TcR-VB gene products . Superantigens can amplify or suppress immune responses . To date, two main groups of superantigens have been described, namely endogenous and exogenous superantigens . Exogenous superantigens are microbial toxins and other protein products . Endogenous superantigens are the products of unlinked genetic loci in mice the best known of which are the murine retroviral gene products . Toxins of S . aureus and S . pyogenes are the best known exogenous superantigens, implicated in Toxic Shock Syndrome.

Mikrobiyol Bul, 1993 Apr, 27(2), 147 - 53
{In vitro sensitivity of Actinobacillus actinomycetemcomitans to various antibiotics}; Akincibay H et al.; In this study, 7 strains of Actinobacillus actinomycetemcomitans (A.a.) were isolated from Localized juvenile periodontitis (LJP) patients and antibiotic susceptibility test by disc diffusion method was utilized . The purpose of this study is to find out the most useful antibiotic against A.a . which is the most important microbial agent in LJP . All seven strains of A.a . were found susceptible to tetracycline, doxycycline, erythromycin, ciprofloxacin and cefuroxime . Two strains were found intermediate susceptible to cephradine . One strain was found resistant to penicillin and another one was found resistant to ampicillin.

J Autoimmun, 1993 Apr, 6(2), 227 - 36
Cellular immune responses and pathogenesis in c-ANCA positive vasculitides; Rasmussen N et al.; Very little is known about the cellular immune response in c-ANCA (classical anti-neutrophil cytoplasm antibodies) positive vasculitides or Wegener's granulomatosis (WG) . The present review is mainly based on published and unpublished observations of the Cattegat Study Group of Wegener's Granulomatosis . Immunohistochemical examinations of nasal biopsies from untreated patients with active WG revealed the presence of substantial amounts of cells belonging to the immune system (CD3+, CD4+, CD8+, CD20+, CD38+ and CD68+) . The IgG response of peripheral mononuclear cells from untreated WG patients to PWM and EBV stimulation was significantly depressed when examined by the reverse haemolytic plaque forming cell assay . T lymphocyte proliferation was observed by in-vitro stimulation with extract of human neutrophil alpha granules, containing the autoantigen for c-ANCA, proteinase-3, only in patients with c-ANCA and active disease . Electron microscopy of biopsies from nasal lesions in untreated patients with active WG did not show structural abnormalities of the organelles of macrophages, giant cells or epithelioid cells . It is concluded that the immune response leading to c-ANCA production involves the full spectrum of immunocompetent cells normally engaged in IgG production to a foreign antigen . This would be in agreement with a theory of cross reactivity of proteinase-3 with a microbial antigen as a pathogenetic mechanism.

Trends Biochem Sci, 1993 Apr, 18(4), 140 - 4
Autoimmune disease and molecular mimicry: an hypothesis; Baum H et al.; Helper T lymphocytes are normally only stimulated to initiate an immune reaction through the recognition of peptides bound to class II major histocompatibility complex (MHC) molecules . Class II MHC molecules are constitutively expressed on antigen-presenting cells which play a critical role in the initiation of immune responses . In disease states, however, other cells often express class II MHC molecules inappropriately . This article suggests an hypothesis for the pathogenesis of autoimmune diseases based on molecular mimicry . The mimicry described is between microbial or viral peptides presented by antigen-presenting cells and self peptides presented inappropriately on a target tissue . This leads to helper T cells, stimulated by peptides derived from infectious organisms, initiating an autoimmune attack on the target tissue.

Inflammation, 1993 Apr, 17(2), 217 - 25
Expression of tumor necrosis factor and c-fos genes in peritoneal macrophages of hypothyroid mice; Liu WK; It is well documented that hypothyroid patients are more susceptible to microbial infections . In order to study whether this impaired response is due to a decrease in production of antitumor molecules or an impaired ability to transmit information in the macrophage, a hypothyroid animal model was used to study the expression of both tumor necrosis factor gene and c-fos protooncogene in peritoneal macrophages . Inbred mice were rendered hypothyroid by an antithyroid drug, methimazole, and the expression of tumor necrosis factor gene and c-fos protooncogene in peritoneal macrophages were studied . Impairment of c-fos and TNF genes were at both transcriptional and translational levels using northern blot analysis, bioassays, and immunocytochemical staining methods . These results indicate that the reduction in signal transduction and in the production of antitumor molecules may cause the poor defense ability of hypothyroid animals.

Can J Vet Res, 1993 Apr, 57(2), 74 - 8
The influence of age and health status on the serum alpha 1-acid glycoprotein level of conventional and specific pathogen-free pigs; Itoh H et al.; Serum alpha-1-acid glycoprotein (alpha 1AG) was measured in 212 Landrace White pigs between birth and finishing age . The alpha 1AG level of healthy pigs five to ten months of age was 338 +/- 79 micrograms/mL, and the upper normal limit in mature swine has been established as 500 micrograms/mL . In both specific pathogen-free (SPF) and conventional pigs, the alpha 1AG level within one day of birth was 14,263 +/- 2,393 micrograms/mL, 40 times the normal adult value, but rapidly decreased to 699 +/- 186 micrograms/mL by four weeks of age . In conventional pigs, alpha 1AG began to increase after four weeks, averaged 1,428 micrograms/mL by eight weeks, but gradually decreased to adult levels by 20 weeks of age . In comparison, alpha 1AG of SPF pigs was only 800 micrograms/mL at eight weeks and decreased more rapidly to normal by 16 weeks of age . The conventional pigs had a high incidence of clinical pneumonia and specific antibodies to Actinobacillus pleuropneumoniae and Mycoplasma hyopneumoniae at the age of eight weeks . As the clinical pneumonia disappeared, serum alpha 1AG level also gradually declined . In contrast, SPF pigs had little clinical illness, low alpha 1AG, and little serological evidence of microbial infection . Conventional pigs with nonrespiratory infections, encephalitis, or with hernias had increased alpha 1AG . While the very high alpha 1AG level of the neonatal pig may be due to genetic influences, increases later in life are likely in response to stimuli from its external environment . Monitoring of serum alpha 1AG in several herds aided in the recognition of disease processes and may have potential use in swine herd health management.

Ann Med, 1993 Apr, 25(2), 171 - 3
Intrauterine contraceptive device and pelvic inflammatory disease; Toivonen J; Comprehensive, mainly epidemiological data has helped us to recognize the strongest risk factors of pelvic inflammatory disease during intrauterine contraceptive device use . These are user's exposure to sexually transmitted diseases, young age and insertion of the device . Moreover, parity of the user and type of device may affect the risk of pelvic infectionPIP: Some people are concerned that IUD use increases the risk of pelvic inflammatory disease (PID), but the extent of the risk and the causal relationship between IUD use and PID are not clear . Two methodological problems with these studies are bias in selecting the comparison group and lack of standard diagnostic criteria for PID . Nonetheless, these studies have identified conditions which increase the risk of PID during IUD use . The woman's or her partner's sexual behavior determines exposure to sexually transmitted diseases (STDs) . Regardless of IUD use, women with at least 4 sexual partners face a 3-fold higher risk of PID than those with only 1 partner . Among IUD users, increased coital frequency (5-8 times/month) increase the risk of PID . IUD insertion introduces temporary microbial contamination into the uterus, thereby bringing about a 6-fold increased risk of PID during the first 20 days after insertion . PID rarely occurs in women who have an IUD for more than 5 years, suggesting the need for less frequent replacement of IUDs . Young age alone is a risk factor for PID . For example, women less than 25 years old face a 2-3 fold increased risk of PID compared to women older than 25 . Nulliparity appears to be a risk factor for PID, but not a strong factor . The Dalkon Shield IUD has been removed from the market because studies indicated that it significantly increased the risk of PID . Copper-releasing IUDs may have a lower risk of PID than does the Dalkon Shield . Studies show that women with the levonorgestrel-releasing IUD have a significantly lower rate of PID than do those with a copper-releasing IUD . Women in a mutually stable relationship are best suited for IUD use . Physicians should diagnose and treat STDs before insertion of an IUD . IUD replacement should be limited, if possible . Women less than 25 years old, particularly those who are nulliparous, should avoid using an IUD .

J Dairy Sci, 1993 Apr, 76(4), 1063 - 73
Computerized monitoring of gas production to measure forage digestion in vitro; Pell AN et al.; The techniques reported in this paper were developed to facilitate the study of the kinetics of forage digestion in vitro by measuring gas production . Fiber disappearance as a measure of the reaction rate has been replaced by the use of computerized pressure sensors to monitor the gaseous products (CO2, CH4) of microbial metabolism . The recording system described requires a computer, pressure sensors, an interface card, and appropriate software to monitor gas production continuously . Several variables, including sample size, inoculum size, vessel size, and type of pressure sensor, have been investigated to determine ranges within which gas production can be measured accurately, and the reproducibility of the results has been established . Because this technique uses small (100-mg) samples, a modified NDF method has been introduced that allows determination of extent of digestion at the end of an incubation in which gas production has been monitored . A strong linear relationship existed between NDF disappearance and gas production.

Biotechnol Appl Biochem, 1993 Apr, 17 ( Pt 2), 251 - 6
Chitinolytic digestibility of cross-linked chitosan gels by an enzyme complex from Streptomyces kurssanovii; Tikhonov VE et al.; The action of a microbial chitinolytic enzyme complex from Streptomyces kurssanovii on chitosan gels with various degrees of cross-linking and on the O-derivatives of the gels was examined . The rate of digestion of the gels was shown to depend on the degree of cross-linking as well as on the size of the O-substituent, decreasing greatly with a rise in substitution.

Transfus Med Rev, 1993 Apr, 7(2), 96 - 103
Challenges in transfusion microbiology; Barbara JA; PIP: Transfusion microbiology must minimize the risks of transfusion transmitted infection (TTI) in a cost-effective and efficient manner . The wide range of transmissible microbial agents exhibits different relative risks in different circumstances; therefore, the same safety rules are not always applicable . TTIs can also exist as asymptomatic diseases in their hosts, so donors must be screened for high-risk behavior . Then blood must be screened by detecting antibodies to infectious agents . Since antibody reactivity is difficult to confirm, and cross-reactive or nonspecific effects are possible, a reactive sample can be tested by a range of assays, each based on a different immunoassay principle, or by a supplementary assay . Despite the advanced nature of these tests, problems of indeterminate donors remain, and a compromise must be made between the sensitivity and specificity of screening assays; maximum sensitivity protects transfusion recipients, but specificity prevents wasting blood or making false reports about donor health . An ideal screening assay would have maximum sensitivity, optimal specificity, simplicity, objectivity, standard format, rapid processing time, safety of reagents, and economy . Assay users must chose the best combination of available characteristics for their particular situation . For example, a French study found that 30% of seronegative, but HIV-infectious, donors are anti-HBc positive; this test would be too specific, however, in a country where hepatitis B is common . Combined assays are being used to screen for anti-HIV-1 and -2 and anti-human T-cell lymphotropic virus-I and -II; this should save time and money . Screening donors selectively (for example, Latin American migrants to North America for Trypanosoma cruzi) may protect previously unexposed populations . In some countries, transmission of HIV via seronegative blood is a problem, and HIV antigen screening and confirmation of reactivity by neutralization should occur as soon as reagents become less expensive . As test methodology continues to move through rapid developmental "generations," the development of sensitive techniques to detect nucleic acids holds promise . Several problems will remain to be solved including cost cross-contamination and carry-over in automated sampling systems, and operational and clerical errors . While there are methods to inactivate viruses in plasma pools, inactivation must incur a minimal loss of biological activity . All of these challenges are exacerbated in developing countries by a lack of resources . As physicians become knowledgeable about when to use transfusion and when to use an alternative method, such as blood salvage, refusing transfusion will be more risky than contracting a TTI .

Ophthalmology, 1993 Apr, 100(4), 452 - 5
Vitreous cultures in suspected endophthalmitis . Biopsy or vitrectomy?
Donahue SP, Kowalski RP, Jewart BH, Friberg TR.
BACKGROUND: Isolation of bacteria from vitreous biopsy often guides therapy in suspected endophthalmitis . Therapeutic vitrectomy provides an additional source of culture material . The authors compared the ability of these two techniques to isolate organisms from patients with acute endophthalmitis . METHODS: In a large ophthalmic microbiology laboratory during a 4-year period, the authors analyzed 206 microbial culture results from patients with suspected endophthalmitis . RESULTS: Two hundred six cases were evaluated . While cultures of vitreous biopsy specimens obtained using a needle and syringe were positive in 91 (53.8%) of 169 patients, culturing the contents of the vitrectomy cassettes produced positive cultures in 29 (74.8%) of 39 patients . Both techniques were performed on 23 patients . Vitreous biopsy allowed isolation of the causative organism in 43% of these patients, whereas vitrectomy was 76% successful . Both comparisons were significant at the P < 0.01 level . No positive vitreous biopsy cultures had associated negative vitrectomy cultures . CONCLUSION: Culturing the contents of the vitrectomy cassette significantly increases the likelihood of obtaining a positive culture compared with merely culturing a vitreous biopsy.

J Anim Sci, 1993 Apr, 71(4), 1019 - 25
Effects of supplemental protein source on passage of nitrogen to the small intestine, nutritional status of pregnant ewes, and wool follicle development of progeny; Schloesser BJ et al.; Rambouillet wethers, surgically fitted with ruminal and duodenal cannulas, were used to determine the influence of substituting soybean meal (SBM) with blood meal (BM) in grass hay diets (8.0% CP) on N flow to the small intestine . Treatments were arranged in a 3 x 4 Latin square design and included diets (10.6% CP) supplemented with SBM; 2/3 SBM:1/3 BM; 1/3 SBM:2/3 BM; or BM . Ruminal NH3 N concentration decreased linearly (P = .03) as BM replaced SBM; however, feeding BM did not affect (P > .10) the total N, microbial N, or nonammonia N flows to the small intestine . A second experiment was conducted to determine the influence of substituting SBM with BM on ewe BW and condition score changes, blood metabolite profiles, wool growth, and progeny wool follicle development . Thirty-two pregnant Targhee ewes fed a grass hay diet were allotted randomly to four groups (n = 8) and supplemented with either SBM; 2/3 SBM:1/3 BM; 1/3 SBM:2/3 BM; or BM . Soybean meal, BM, or their combinations provided 22 g of dietary CP daily . Dietary treatments had no influence on ewe BW (P = .40) or body condition score (P = .77) changes, and ewes gained 7.6 kg of BW during the 84-d experiment . Protein source did not influence (P > .10) serum protein, urea N, creatinine, glucose, or nonesterified fatty acid concentrations . Lamb wool follicle density per square millimeter and secondary:primary follicle ratios were 22.4 and 10.8, respectively, and were not different (P > .10) among dietary treatments . Compared with SBM, no advantages were detected for BM supplementation of grass hay diets in these studies.

Appl Environ Microbiol, 1993 Apr, 59(4), 1242 - 6
Biodegradability and mechanical properties of poly-(beta-hydroxybutyrate-co-beta-hydroxyvalerate)-starch blends; Ramsay BA et al.; Wheat starch granules and poly-(beta-hydroxybutyrate-co-beta-hydroxyvalerate) {P(HB-co-HV), (19.1 mol% HV)} were blended at 160 degrees C . Increasing the starch content from 0 to 50% (wt/wt) decreased the tensile strength of P(HB-co-HV) from 18 MPa to 8 MPa and diminished flexibility as Young's modulus increased from 1,525 MPa to 2,498 MPa, but overall mechanical properties of the polymer remained in a useful range . A mixed microbial culture required more than 20 days to degrade 150-microns-thick samples of 100% P(HB-co-HV), whereas samples containing 50% (wt/wt) starch disappeared in fewer than 8 days . Starch granules degraded before P(HB-co-HV) did . Aerobic degradation proceeded more rapidly than anaerobic degradation.

J Prosthet Dent, 1993 Apr, 69(4), 406 - 15
Biocompatibility: its future in prosthodontic research; Edgerton M et al.; The future of prosthodontic research will involve replacing lost tissues by using scientific methods that evaluate biomaterials and treatment designs based on desired biologic outcomes . The present concept of a biocompatible material is one that elicits an appropriate host response in a specific application . To design optimal biomaterials, three interactive components should be considered: the chemical nature of the surface, the mediating pellicle layer, and microbial and host response . Surface chemistry determines which molecules are selectively absorbed onto a surface from oral fluids . The pellicle-coated surface should be designed to elicit a more desirable host response . Pellicle composition can be altered by chemically changing the surface, precoating surfaces with biological molecules, or using synthetic materials designed to mimic natural tissues . Several surface-sensitive techniques are available to assess these modifications, including vibrational spectroscopy, electron microscopy for chemical analysis, and bioanalytical methods . To develop more biocompatible materials, a further understanding of pellicle formation as a function of surface composition, microbial adhesion to biomaterials, and cellular reaction to implant biomaterials is necessary . This knowledge will facilitate development of new biologically based rationales for treatment modalities in restorative dentistry.

J Clin Pharm Ther, 1993 Apr, 18(2), 133 - 7
Decontamination methods for cytotoxic drugs . 1 . Use of a bioluminescent technique to monitor the inactivation of methotrexate with chlorine-based agents; Wren AE et al.; A new microbial bioluminescence assay has been used to monitor the loss of mutagenicity on inactivation of methotrexate by active chlorine-based agents . The drug was degraded to products that were non-active in this mutagen detection system, in agreement with previously described work . Presept granules appear to be a suitable alternative to sodium hypochlorite for inactivating solutions and surface spills of methotrexate . The bioluminescence assay appears to have potential for monitoring clean-up and decontamination procedures in areas where cytotoxic agents are used.

J Anim Sci, 1993 Apr, 71(4), 1057 - 64
Combinations of starch and digestible fiber in supplements for steers consuming a low-quality bromegrass hay diet; Grigsby KN et al.; Five steers (690 kg) with ruminal and duodenal cannulas were fed 100% bromegrass hay or 60% bromegrass hay:40% concentrate at 90% of their ad libitum DMI . The concentrate consisted of ground corn (GC) and(or) soybean hulls (SBH): 1) 100% SBH; 2) 66% SBH/33% GC; 3) 33% SBH/66% GC; or 4) 100% GC . Increasing level of corn inclusion decreased (linear, P < .01) DM and OM digestion (corrected for microbial DM and OM flow) in the rumen . Neutral detergent fiber digestion decreased in the rumen (quadratic, P = .05) and total tract (linear, P = .06) with increasing level of corn . Ruminal NDF digestion (quadratic, P = .05) and duodenal microbial N flow (quadratic, P < .01) were greatest for 66% SBH/33% GC, whereas total N flow to the duodenum was similar (P > .10) among treatments . Microbial efficiency increased (quadratic, P = .10) as the level of corn inclusion increased . Molar percentage of ruminal acetate decreased (linear, P < .01) with increasing level of corn, whereas molar percentage of butyrate increased (linear, P < .10) . Carboxymethylcellulase activity was greatest (quadratic, P = .08) for 66% SBH/33% GC and paralleled NDF digestion in the rumen . Increasing level of corn decreased (linear, P < .07) in situ DM digestibility of SBH after 4, 8, and 12 h of incubation.

Scand J Immunol, 1993 Apr, 37(4), 509 - 14
IgA and IgG subclass deficiency in a poor population in a developing country; Castrignano SB et al.; The levels of IgG, IgG subclasses, IgM and IgA were determined in serum from 17 patients with IgA deficiency and severe or frequent infections, allergy and/or autoimmunity (median age 7 years, range 2-19), 11 healthy IgA-deficient adults and 35 controls (median age 7 years, range 2-19) . In serum from all groups IgG, IgM and IgA antibodies were determined against beta-lactoglobulin, E . coli O antigens and poliovirus type 1 antigen . In saliva of 15 IgA-deficient patients and 12 of the controls IgG, IgM and secretory component-carrying antibodies against E . coli O antigens and poliovirus type 1 were determined . The majority of the studied individuals lived under poor socio-economic conditions in Brazil, with consequent heavy microbial exposure . One IgA-deficient patient with rheumatoid arthritis also had IgG2 deficiency but no infectious problems . Four out of the 35 controls without any obvious infectious problems were found with IgA or IgG subclass deficiency . One of the 11 healthy IgA-deficient adults was low in the IgG2 subclass, one in IgG1 and one in IgG3 . Those with symptomatic IgA deficiency had significantly higher serum IgG than the controls, especially in the age group 6-11 years . This latter group also had significantly increased serum IgG1 and IgG2 levels when compared with the age-matched controls . Salivary IgM antibodies to E . coli and poliovirus antigens were significantly higher among the symptomatic IgA-deficient individuals than among the controls . It is not clear at present whether these increased Ig levels are secondary to frequent infections and/or part of mechanisms that may compensate for the IgA deficiency.

J Reprod Immunol, 1993 Apr, 23(3), 223 - 33
An immunohistologic analysis of murine uterine T cells between birth and puberty; Croy BA et al.; Murine uterine T cells were analysed on the basis of surface phenotype expression from birth to adulthood . T cells were rare in the uterus from birth until 2 weeks of age . In genetically immunocompetent mice, mature T cells expressing either TCR alpha/beta or TCR gamma/delta were first present as a major cell population at 3 weeks of age . The ratio of TCR alpha/beta to TCR gamma/delta was 1:1 at 3 weeks of age and this ratio did not change during sexual maturation . Almost all uterine T cells were CD8+ and the majority of these cells expressed CD8 alpha/beta rather than CD8 alpha/alpha . Cells expressing Thy1.2 were less frequent than cells expressing CD3 while cells expressing CD5 were rare . No major changes in T cell subsets occurred at puberty . Further, the microbial flora of the mice did not alter the time of appearance, frequency or subset distribution of uterine TCR+ cells . In the uteri of immunodeficient mice of genotype scid/scid TCR+ cells were found in low numbers and the initial appearance of TCR+ cells was delayed until 5 weeks of age.

Zentralbl Bakteriol, 1993 Apr, 278(2-3), 377 - 82
Possible role of bacterial heat-shock proteins in autoimmune diseases; van Eden WV; The immune system is continuously exposed to a variety of microbial antigens . It is unavoidable that such antigens may resemble autoantigens, being present in the host . It has appeared now that the immune system is accepting recognition of such "mimicry" antigens and even may have developed a tendency to focus its attention to such antigens . This paper discusses the findings of immune responses to heat-shock proteins, as a family of proteins exhibiting extraordinary sequence conservation, creating a high degree of similarity between bacterial antigens and host antigens . On the basis of existing evidence, it is argued that immune responses directed at hsps are part of regulatory mechanisms which enable the immune system to safely contain potentially self-reactive lymphocytes . Furthermore, it is argued that by means of artificial immunization against hsps or selected determinants of hsps, a development of autoimmune diseases may be inhibited.

Zentralbl Bakteriol, 1993 Apr, 278(2-3), 275 - 86
Variation and genetic control of surface antigen expression in mycoplasmas: the Vlp system of Mycoplasma hyorhinis; Yogev D et al.; Surface antigenic diversity in the swine pathogen Mycoplasma hyorhinis is generated by random combinatorial expression and high-frequency phase variation of multiple, size-variant membrane surface lipoproteins (Vlps) which represent the major coat proteins of this wall-less procaryote . The distinctive structural basis for Vlp variation was revealed in a family of several related but divergent vlp genes . These occur in one cluster as single chromosomal copies, each encoding a conserved domain for membrane insertion and lipoprotein processing, and a divergent external domain that changes size by deletion or insertion of repetitive intragenic coding sequences while retaining a distinctive charge motif . Lack of detectable changes in restriction fragment patterns or DNA sequence of vlp structural genes during phase transitions between ON and OFF expression states ruled out long range genomic rearrangements and frameshift mutations as a means of controlling Vlp phase variation . However, highly homologous vlp promoter regions contain a homopolymeric tract of contiguous adenine residues {poly(A)} upstream of the transcriptional start site which is subject to frequent mutations altering its length . These mutations are the only sequence changes detected during phase transitions, and are highly correlated with the expression state of each vlp gene . This suggests a mechanism of transcriptional control regulating Vlp phase variation by critical changes within the poly(A) region affecting the spacing between the -10 and -35 hexamers or a putative regulator binding site . The multiple levels of structural and antigenic diversity embodied in the vlp gene family may provide essential adaptive capabilities for this wall-less microbial pathogen.

Immun Infekt, 1993 Apr, 21 Suppl 1, 31 - 3
{The mucosa-associated immune system--immune defense in the gastrointestinal tract}; Schreiber S et al.; The intestinal immune system has to protect the mucosa against microbial and toxic agents . This is achieved by elimination of potential antigens, which are inhibited to adhere and to invade the mucosa . Thus, induction of inflammatory processes is prevented . Elimination of antigens is mediated by the secretion and transport of IgA (and especially IgA2) to the lumen of the gut . There, IgA is the most efficient part of the so-called "mucosa block" . The composition of mononuclear cells in the lamina propria exhibits the preference of a memory T cell type that is able to provide help for B cells . If IgA-mediated immune response proves not to be sufficient, IgG-mediated inflammatory processes are induced within the mucosa . The role of cytotoxic T cells and, moreover, the biological function of intraepithelial lymphocytes are still controversial.

Semin Cell Biol, 1993 Apr, 4(2), 93 - 102
Cell wall carbohydrates as signals in plants; Mohnen D et al.; Plant and fungal cells are surrounded by a cell wall rich in diverse polysaccharides and proteins . It has become apparent in recent years that the carbohydrates in the cell wall function not only to maintain cell shape and integrity, but also may serve as signals in plants . This review summarizes the evidence that biologically-active oligosaccharides (oligosaccharins) released from plant or microbial cell walls can serve as signals to regulate plant defense and plant growth and development . The oligosaccharins discussed include the fungal-derived hepta-beta-glucoside and the plant cell wall-derived oligogalacturonides and xyloglucans . Possible mechanisms by which oligosaccharins may exert their effects on plant cells are discussed.

Plant Physiol, 1993 Apr, 101(4), 1239 - 47
Posttranslational processing of a new class of hydroxyproline-containing proteins . Prolyl hydroxylation and C-terminal cleavage of tobacco (Nicotiana tabacum) vacuolar chitinase; Sticher L et al.; The fungicidal class I chitinases (EC 3.2.1.14) are believed to be important in defending plants against microbial pathogens . The vacuolar isoforms of tobacco (Nicotiana tabacum), chitinases A and B, are the first examples of a new type of hydroxyproline-containing protein with intracellular location, enzymic activity, and a small number of hydroxyprolyl residues restricted to a single, short peptide sequence . We have investigated the posttranslational processing and intracellular transport of transgene-encoded chitinase A in callus cultures of Nicotiana tabacum L . cv Havana 425 and leaves of Nicotiana sylvestris Spegazzini and Comes . Pulse-chase experiments and cell fractionation show that chitinase A is processed in two distinct steps . In the first step, the nascent protein undergoes an increase in apparent M(r) of approximately 1500 detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Experiments with the inhibitor of prolyl hydroxylation, alpha,alpha'-dipyridyl, and pulse-chase labeling of cells expressing recombinant forms of chitinase A indicate that the anomalous increase in M(r) is due to hydroxylation of prolyl residues . This step occurs in the endomembrane system before sorting for secretion and vacuolar transport and does not appear to be required for correct targeting of chitinase A to the vacuole . The second step is a proteolytic cleavage . Sequencing of tryptic peptides of the mature proteins indicates that during processing essentially all molecules of chitinase A and B lose a C-terminal heptapeptide, which has been shown to be a vacuolar targeting signal . This appears to occur primarily in the endomembrane system late in intracellular transport . A model for the posttranslational modification of chitinase A is proposed.

Microb Pathog, 1993 Apr, 14(4), 287 - 97
Immune recognition of human Hsp60 by Lyme disease patient sera; Girouard L et al.; Members of the Hsp60 family of microbial heat shock proteins frequently serve as immunodominant antigens and immunological responses to these highly conserved proteins have been implicated in the pathology of a number of autoimmune diseases and inflammatory processes associated with microbial infection . In the present study, sera from patients with Lyme disease were examined by Western blot analysis for the presence of IgG against Borrelia burgdorferi antigens and for autoreactive IgG against recombinant human Hsp60 (huHsp60) . These results were then compared to those obtained using sera from normal healthy controls, patients with a variety of acute non-spirochete infections, and patients with rheumatoid arthritis (RA) . The results indicate a high incidence of autoreactive antibodies against huHsp60 in the sera of Lyme disease patients (67.9%) and patients with RA (75%) . Positive reactivity was observed at lower rates in sera from healthy controls (25%) and sera from patients with acute non-spirochete infections (38%) . Together the data suggest an association between the presence of autoreactive antibodies against huHsp60 and infection with B . burgdorferi . A similar association may exist between the presence of autoreactive antibodies against huHsp60 and RA.

Appl Microbiol Biotechnol, 1993 Apr, 39(1), 26 - 30
Characteristics of cross-flow filtration of pullulan broth; Yamasaki H et al.; Cross-flow filtration of culture broth from Aureobasidium pullulans, which elaborates pullulan, was done with a thin channel-type module and microfiltration membranes made of different materials and with different pore sizes . Various factors affecting the results of the filtration were studied . The specific resistance of the microbial cake was found to be higher than that of bakers' yeast, the cells of which are about the same size as an A . pullulans cell, and resistance increased with cultivation time . The flux and transmission of pullulan through the membrane decreased with cultivation time as the specific resistance increased . The flux and transmission of pullulan depended on the structure and pore size of the membrane and also on the pH of the broth . With a polysulphone membrane with a nominal pore size of 2.0 microns, transmission was nearly 100% with negligible leakage of cells and the flux was high when the pH of the broth was adjusted to 2.0.

FEBS Lett, 1993 Mar 22, 319(3), 253 - 6
Peptide aldehydes as inhibitors of HIV protease; Sarubbi E et al.; We have recently shown that alpha-MAPI, a peptidic aldehyde of microbial origin, inhibits the HIV protease with a potency comparable to pepstatin, having, differently from pepstatin, no activity on other aspartic proteases . In this study different peptide derivatives containing a C-terminal aldehyde have been tested to assess the potential of this function for the inhibition of HIV protease . The results of our analysis correspond with the recently published subsite preferences of the viral enzyme, indicating that aldehydes bind to the active site of the HIV protease . Our data suggest that peptide aldehydes can act in their hydrated forms as transition state analogues with the most potent inhibitor having an IC50 of 0.9 microM.

Proc Natl Acad Sci U S A, 1993 Mar 15, 90(6), 2281 - 5
Identification of Ras farnesyltransferase inhibitors by microbial screening; Hara M et al.; A microbial screen using a yeast strain with conditional deficiency in the GPA1 gene was carried out to search for inhibitors of protein farnesyltransferase (PFT) . A strain of Streptomyces was found to produce active compounds named UCF1-A, UCF1-B, and UCF1-C . Structural determination of these compounds revealed that UCF1-C is identical to the known antibiotic, manumycin, whereas UCF1-A and UCF1-B are structurally related to manumycin . All three UCF1 compounds suppress the lethality of gpa1 disruption, with UCF1-C exhibiting the strongest activity . UCF1 inhibits yeast as well as rat brain PFT . Fifty percent inhibition of yeast PFT activity is observed with 5 microM UCF1-C . Kinetic analyses of the inhibition suggest that UCF1-C acts as a competitive inhibitor of PFT with respect to farnesyl pyrophosphate, exhibiting a Ki of 1.2 microM, whereas the same compound appears to act as a noncompetitive inhibitor of PFT with respect to the farnesyl acceptor, the Ras protein . UCF1-C shows significant activity to inhibit the growth of Ki-ras-transformed fibrosarcoma, raising the possibility of its use as an antitumor drug.

J Immunol Methods, 1993 Mar 15, 160(1), 65 - 71
A sensitive method to detect defined peptide among those eluted from murine MHC class II molecules; Brumeanu TD et al.; We developed a sensitive competitive inhibition radioimmunoassay able to trace pmoles of a defined peptide eluted from major histocompatibility complex (MHC) class II molecules that were subsequently fractionated by RP-HPLC . In this assay we used a model synthetic peptide corresponding to amino acid residues 110-120 from the hemagglutinin (HA) of PR8 influenza virus, and affinity purified rabbit antibodies specific for this peptide . The HA110-120 peptide binds to I-Ed class II molecules on the surface of APCs and is recognized by specific CD4+ T helper cells . 2PK3 B lymphoma cells (H-2d) were pulsed with HA110-120 peptide or PR8 virus, lysed, the MHC class II molecules extracted, and bound peptides eluted . After separation by RP-HPLC, the fractions were tested for inhibition of the binding of rabbit anti-HA110-120 antibodies to peptide coated microtiter plates . A significant inhibitory activity was observed with one peak when the cells were pulsed with HA110-120 peptide and two peaks when pulsed with PR8 virus . The inhibitory activity was correlated with the presence of HA110-120 peptide as demonstrated by peptide sequencing . The assay is reproducible and sensitive to 1 pmol of antigenic peptide . This assay can be useful to identify microbial peptides with defined structure and antigenicity among the multiple peptides bound to class II molecules.

Mol Cell Biochem, 1993 Mar 10, 120(1), 1 - 13
An allometric interpretation of the spatio-temporal organization of molecular and cellular processes; Aon MA et al.; Different levels of organization distinguished by characteristics spatial dimensions, Ec, and relaxation times, Tr, of biological processes ranging from electron transport in energy transduction to growth of microbial and plant cells, are shown to be related through a relation that may be interpreted as allometric and characterized by two different slopes . Processes, at levels of organization occurring in spatial dimensions of micrometers and relaxing in the order of minutes, delimit a 'transition point' between the two curves, that we interpret as a limit for the emergence of macroscopic coherence . The characteristic spatial dimension, Ec, and the relaxation time, Tr, contain dynamical information about the processes occurring at a given level of organization . When a steady state of a biological process at a certain level of organization becomes unstable, the system undergoes a transition to another level of organization . To exemplify the appearance of macroscopic order at levels of organization further from the 'transition point' we present in this report various experimental systems involving many levels of organization allometrically related that exhibit different kinds of self-organized behavior, i.e . bi-stability, oscillations, changes in (a)symmetry.

Med Hypotheses, 1993 Mar, 40(3), 182 - 5
Zinc in etiology of periodontal disease; Polenik P; Microbial plaque is the main etiological factor of periodontal disease . The bacterial polysaccharides stimulate gingival neutrophils and macrophages to interleukin-1 (IL-1) production . IL-1 causes a complex of redistribution processes with liver as the central organ . Accumulation of zinc in liver and their copper and ceruloplasmin production also elicits increase of copper and decrease of zinc in gingiva . The elevated level of copper in connection with zinc deficiency in gingiva causes the increase of permeability of gingival epithelium for bacteria . The stimulated inflammatory infiltrate produces more IL-1 and the vicious circle is complete.

Dev Comp Immunol, 1993 Mar-Apr, 17(2), 97 - 108
In vitro phenoloxidase activity in the blood of Ciona intestinalis and other ascidians; Jackson AD et al.; The presence and activation of phenoloxidase in the blood of Ciona intestinalis and other ascidians was investigated in vitro . In C . intestinalis, phenoloxidase was found to exist in the cells as a proenzyme and to be activated by proteases . The microbial carbohydrates, LPS or laminarin, also enhanced enzyme activity but a similar effect was not achieved with other sugars . Calcium was not essential for enzyme activity and no enzyme suppression was seen at high calcium concentrations . Prophenoloxidase activation by LPS was further found to be dose related and inhibited by PTU and tropolone . Since benzamidine and STI reduced phenoloxidase activity in cell lysate supernatants, activation may involve other factors, possibly a serine protease . Lastly, as phenoloxidase activity was detected in the blood cells (usually the morula cells) of eight other ascidian species, it appears that it is widely distributed in the blood of this group of invertebrates.

Mol Microbiol, 1993 Mar, 7(6), 905 - 14
Molecular characterization of the enniatin synthetase gene encoding a multifunctional enzyme catalysing N-methyldepsipeptide formation in Fusarium scirpi; Haese A et al.; The gene encoding the multifunctional enzyme enniatin synthetase from Fusarium scirpi (esyn1) was isolated and characterized by transcriptional mapping and expression studies in Escherichia coli . This is the first example of a gene encoding an N-methyl peptide synthetase . The nucleotide sequence revealed an open reading frame of 9393 bp encoding a protein of 3131 amino acids (M(r) 346,900) . Two domains designated EA and EB within the protein were identified which share similarity to each other and to microbial peptide synthetase domains . In contrast to the N-terminal domain EA, the carboxyl terminal domain EB is interrupted by a 434-amino-acid portion which shows local similarity to a motif apparently conserved within adenine and cytosine RNA and DNA methyltransferases and therefore seems to harbour the N-methyl-transferase function of the multienzyme.

J Obstet Gynecol Neonatal Nurs, 1993 Mar-Apr, 22(2), 169 - 79
Pelvic inflammatory disease: risk factors and microbial etiologies; Jossens MO et al.; OBJECTIVE: To review the risk factors and microbial etiologies of pelvic inflammatory disease (PID) . DATA SOURCES: Include 77 current and historical references on PID, PID risk factors, and sexually transmitted diseases . STUDY SELECTION: 34 studies that address specific research questions . DATA EXTRACTION: Data were used from studies whose methodologies are discussed . DATA SYNTHESIS: This review identifies research problems regarding the definition, diagnosis, and identification of microbial profiles and risk factors of PID . CONCLUSIONS: More reliable diagnostic criteria for the varied clinical presentations of this polymicrobial infection are needed . PID is a disease with multiple, interrelated risks . Education is an important nursing intervention.

Chem Pharm Bull (Tokyo), 1993 Mar, 41(3), 611 - 3
Microbial hydroxylation of (-)-eburnamonine by Mucor circinelloides and Streptomyces violens; Adachi T et al.; (6R)-6-Hydroxy-, (6S)-6-hydroxy- and (18S)-18-hydroxyeburnamonines were obtained by microbial conversion of (-)-eburnamonine using Mucor circinelloides and Streptomyces violens . Their structures were determined by analyses of the mass, 1H- and 13C-NMR spectra . (-)-Eburnamonine and the three hydroxylated compounds showed cerebral protecting effects against potassium cyanide intoxication in mice.

J Dairy Sci, 1993 Mar, 76(3), 677 - 90
Release of volatile branched-chain and other fatty acids from ruminant milk fats by various lipases; Ha JK et al.; Bovine, ovine, and caprine milk fats were treated with pregastric lipases (kid goat, calf, and lamb), microbial lipases (Candida cylindracea, C . cylindracea AY30, Aspergillus niger APF12, Rhizopus arrhizus, Penicillium roqueforti R10, and Mucor zavanicus Map 10), porcine pancreatic lipase, or milk lipase . All three pregastric lipases preferentially hydrolyzed volatile branched-chain and short n-chain fatty acids from each milk fat . Pregastric lipases also released a relatively low proportion of C10 from bovine milk fats but a high proportion of C10 from caprine milk fat . Milk lipase released very low concentrations of butanoic acid and did not release 4-methyloctanoic acid in significant amounts except from caprine milk fat . Ovine milk fat yielded a substantially greater concentration of butanoic acid than did bovine or caprine milk fats when it was hydrolyzed by porcine pancreatic lipase . Candida cylindracea lipase yielded high amounts of volatile n-chain fatty acids nonselectively and only small quantities of volatile branched-chain fatty acids . High amounts of the medium-chain branched fatty acids were produced by kid goat, P . roqueforti, A . niger, and R . arrhizus lipases.

Fertil Steril, 1993 Mar, 59(3), 617 - 28
The pH as an important determinant of sperm-mucus interaction; Eggert-Kruse W et al.; OBJECTIVE: To determine the clinical significance of endocervical mucus pH on sperm-mucus interaction during infertility investigation . PATIENTS AND MATERIAL: Two hundred sixteen couples with a median duration of infertility of 4 years (range, 1 to 19 years) presenting at the infertility unit of the Women's University Hospital of Heidelberg, Germany . MAIN OUTCOME MEASURES: Determination of endocervical pH by colorimetric and electrometric measurement and correlation of results with the outcome of postcoital testing (PCT) and other parameters of infertility investigation (semen and cervical mucus {CM} quality, microbial colonization of cervix and ejaculates, medical history, hormonal status, and specific medication) and the subsequent fertility in a prospective study . In vitro experiments with the sperm-cervical mucus penetration test (SCMPT) used as biological model . RESULTS: The colorimetric determination of endocervical mucus pH is an easy method, suitable for routine clinical use, correlating significantly with electrometric measurement of pH . Median pH was 7.0 (range, 5.4 to 8.2) . The mucus pH was significantly related with the results of PCT, even when mucus and semen variables were taken into account . No significant relationship was seen between the cervical index and mucus pH and the microbial colonization of cervix and ejaculates . The pH of endocervical secretions correlated with the peripheral hormonal status: low pH levels were significantly more frequent in patients with hyperandrogenemia, indicated by high testosterone and/or dehydroepiandrosterone sulfate levels before medication was started, and in hyperandrogenemic patients treated with dexamethasone than in the other women . Oral administration of estrogens led to a subtle alkalinization of the CM . With regard to subsequent fertility 6 months after pH testing, the pregnancy rate was significantly lower in women offering reduced mucus pH on occasion of the PCT in the group of couples with primary infertility and in couples with oligozoospermia of the male partner . The significant influence of pH on sperm-mucus interaction was confirmed in vitro with the SCMPT . CONCLUSIONS: The results indicate that the pH of the CM, easily determined with pH indicator paper, is an important parameter of mucus quality with significant influence on spermatozoal viability in CM, which correlates with peripheral hormonal status and can be affected by oral medication with estrogens . Therefore the routine determination of pH on occasion of the PCT is recommended during infertility investigation.

Eur J Clin Nutr, 1993 Mar, 47(3), 160 - 73
The Kingston Project . II . The effects of high energy supplement and metronidazole on malnourished children rehabilitated in the community: anthropometry; Heikens GT et al.; Malnourished children (mean age 1.2 years) referred from public health clinics to a paediatric metabolic ward in Kingston, Jamaica, were enrolled for treatment in a community-based health care project and were randomly allocated to one of two groups . The first group was treated at home with metronidazole and then for 6 months using the standard health care provided from local clinics by community health aides . The second group was given the same drug and home treatment, but in addition received a high energy supplement of 3.31 MJ daily for 3 months . We have previously shown a significant advantage in both weight and height gain for a group given the same supplement in contrast with standard health care controls (Heikens et al., 1989, Eur . J . Clin . Nutr . 43, 145-160), and in this study test the addition of a drug treatment aimed at reducing malabsorbtion due to a possible microbial overgrowth of the small bowel in malnourished children . This paper reports anthropometric findings showing significant benefits from both the drug and nutritional treatments . Greatest gains were by the group given both treatments, but the group given the antibiotic treatment, without energy supplementation, also made better growth recovery than did controls . Only 8% of the children treated with metronidazole failed to respond to community-based intervention and were admitted to hospital, compared with 19% for the other groups (P < 0.05) . These findings support targetted high-energy supplementation for the rehabilitation of moderately malnourished children receiving health clinic care, and suggest further that such programs should include antibiotic treatment directed at SBBOPIP: In 1985-86 in Jamaica, a community-based health care project randomly allocated 81 3-36 month old malnourished children from the slums of metropolitan Kingston to either a group receiving home health care/clinic-based care and a 5-day course of a broad spectrum antibiotic, metronidazole (20 mg/kg/day), for 6 months or a group receiving these same interventions and a high energy supplement (HES) (790 kcal) containing 20.6 gm protein for 3 months to test the effect of these interventions on anthropometric measures of growth . The researchers also wanted to determine whether metronidazole would overcome malabsorption of nutrients due to small bowel bacterial overgrowth . Children of both groups benefited considerably from the interventions . For example, significant improvements in weight occurred almost immediately followed by improvements in length, resulting in a significant improvement in the body mass index (BMI) (p = .0001) . Children receiving both HES and metronidazole made significantly greater gains than those who only received HES (weight, p = .02; length, p = .0002; and BMI, p = .0001) . A significantly greater proportion of children did not respond to treatment and had to be hospitalized for infections, especially respiratory infections, in the HES only group than did those in the HES and metronidazole group (19% vs . 8%; p .05) . None of the children receiving metronidazole died . Reduced morbidity, absence of case fatalities, and anthropometric improvements support the belief that home visits by community health aides in combination with clinic-based health services providing HES and antibiotic treatment to moderately malnourished children can indeed rehabilitate them .

J Prosthet Dent, 1993 Mar, 69(3), 314 - 7
Denture adhesives: cytotoxicity, microbial contamination, and formaldehyde content; Ekstrand K et al.; Denture adhesives are extensively used to enhance retention of dentures . Denture adhesives and their leachable components are ingested, but reports on the biologic aspects of denture adhesives are scarce . This study investigated some biological properties of 19 commercially available denture adhesives . The adhesives were assessed by the agar overlay technique and analyzed for microbial contamination and formaldehyde content . In the agar overlay test, all of the materials caused severe cytotoxic effects . Sabouraud dextrose agar was used for cultivation of fungi and tryptone soya broth for cultivation of aerobes, including fungi . Most of the samples tested showed microbial growth . The formaldehyde test revealed the presence of substantial amounts in four products and minor amounts in two products.

J Invest Dermatol, 1993 Mar, 100(3), 225 - 8
A potential role for superantigens in the pathogenesis of psoriasis; Leung DY et al.; Psoriasis is a complex inflammatory skin disease in which local vascular changes, T-cell activation, abnormal keratinocyte proliferation and differentiation, and neutrophil activation all contribute to the ongoing disease process . Because of recent interest in T-cell activation as a trigger for psoriatic lesions, we hypothesized that psoriasis may be triggered by superantigens, e.g., toxins of microbial origin that stimulate T cells expressing particular T-cell receptor (TCR) beta chain variable (V beta) gene segments . Lesional skin biopsies and peripheral blood from two patients with acute exacerbations of their psoriasis that appeared to be triggered by infection were analyzed for TCR V beta gene expression using monoclonal antibodies directed against V beta 5.1, 5.2, 6.7, 8.1, and 12 . Skin biopsies from both patients demonstrated a different pattern of V beta expansion that correspond to the V beta pattern expected to be induced by the type of superantigen expressed during the infection . In contrast, using immunofluorescence and flow cytometry, peripheral blood T cells from these patients did not demonstrate any expansion of the 5 V beta subsets studied . These observations support the hypothesis that local activation of cutaneous T cells in psoriasis may be caused by a superantigen and provides a new direction for investigating the pathogenesis of this complex and fascinating skin disorder.

J Clin Periodontol, 1993 Mar, 20(3), 179 - 85
Gingivitis in young and old dogs; Berglundh T et al.; The aim of the investigation was to study the development of gingivitis in young and old dogs . 10 beagle dogs were used . 5 of the dogs were 1 year old and 5 dogs were between 8 and 9 years of age . On day 0, all the teeth of all 10 dogs were scaled and polished . A 6-week period of plaque control was initiated . On day 42, a clinical examination was performed, microbial samples from the gingival sulcus were harvested and biopsies obtained from tooth regions 3P and 4P . The tooth-cleaning measures were abandoned following biopsy and the animals were during the subsequent 3 weeks allowed to form plaque . On day 63, i.e., day 21 of plaque formation, the clinical examination, sampling of the subgingival plaque and harvesting of biopsies were repeated in the contralateral premolar regions (P3 and P4) . The histological analysis revealed that the cellular infiltrate (the lesion) of inflamed gingiva was larger and extended more apically in the old than in the young dogs . The lesion in the old dogs contained significantly more plasma cells but less macrophages, lymphocytes and PMN cells than the infiltrate in the young dogs.

Clin Exp Immunol, 1993 Mar, 91(3), 482 - 8
An improved enrichment method for functionally competent, highly purified peripheral blood dendritic cells and its application to HIV-infected blood samples; Karhumaki E et al.; Dendritic cells (DC) were purified from human peripheral blood using a rapid and simple method based on magnetic depletion of phagocytes with carbonyl iron, followed by centrifugation of nonphagocytic cells on a Percoll density gradient and depletion of lymphocytes and macrophages/monocytes with a panel of MoAbs and immunomagnetic beads . Enriched DC were obtained with > 99% purity as judged by non-specific esterase (NSE) staining . After isolation, these cells, representing 0.4% of the starting mononuclear cell population, still function as potent antigen-presenting cells for purified T lymphocytes . The present results confirm the ability of human peripheral blood DC to present soluble antigens to T cells including microbial antigens and show, further, that DC are more potent soluble antigen-presenting cells than monocytes . The method was successfully applied to the purification of DC from the blood of HIV-infected individuals . We could not detect decreased numbers of DC in four individuals with early HIV infection and no replicating HIV was detected by in situ hybridization in the DC.

Minerva Gastroenterol Dietol, 1993 Mar, 39(1), 37 - 9
{Purified human IgG administration, after chemotherapy, to patients with colorectal carcinoma}; Faraone V et al.; The authors, in the present study try to test the prophylactic effect of human IgG on surgical patients affected by colorectal cancer and undergoing chemotherapy in order to prevent microbial infections, supporting immunological host defences.

Med J Malaysia, 1993 Mar, 48(1), 9 - 11
Cytokines in infection; Pang T; Cytokines are central to the development of effective immunity against microbial pathogens and their beneficial effects in the control of a variety of infections of man and animals are well-documented . However, it appears that cytokines may also have detrimental effects in infections by actually enhancing microbial growth . These observations emphasise the fact that many successful pathogens possess mechanisms enabling them to evade the immune response and that caution needs to be exercised in using cytokines as therapeutic agents to control infections in various human diseases.

East Afr Med J, 1993 Mar, 70(3), 182 - 5
Biocidal effects and residual glutaraldehyde in cellulosic and synthetic polymers; Mwaniki DL et al.; The amount of leachable glutaraldehyde from medical grade Cuprophan 150PM (cellulose based) and AN69S (a copolymer of acrylonitrile and sodium methallyl sulphonate) membranes, and polyproprene plates following 72 hr immersion in 2% alkaline glutaraldehyde was determined . After rinsing the materials for 30 minutes in distilled water at 37 degrees C, the mean concentration of leachable glutaraldehyde from Cuprophan 150 PM (8 . 60 +/- 0.55 micrograms/g) was significantly greater than that from AN69S membrane (6.50 +/- 0.60 micrograms/g), (p < 0.01) . The mean leachable glutaraldehyde from plates was 0.30 +/- 0.15 microgram/g . There was a significant decrease in the mean leachable glutaraldehyde from AN69S (5.35 +/- 0.25 micrograms/g) after second immersion (p < 0.05) . The concentration of leachable glutaraldehyde in the plates and Cuprophan 150PM remained relatively unchanged . Absence of growth following infusion of 2% alkaline glutaraldehyde into blood contaminated miniature dialyzers containing high microbial loads of selected bioindicator organisms (S . aureus, P . aeruginosa, B . subtilis (Var globigii) spores, M . gordonae, A . niger spores and attenuated polio virus) demonstrated its effectiveness under environmental conditions that were conducive to high resistance . These findings indicate that 2% alkaline glutaraldehyde is readily washed from the three polymer materials that were studied and its accumulation following repeated exposure was not evident . Its application in pre-sterilization treatment of heavily contaminated invasive polymer based devices seemed feasible.

Res Immunol, 1993 Mar-Apr, 144(3), 198 - 201; discussion 214-22
Roles of superantigens in microbial infections?
Acha-Orbea H.
Superantigens have been defined in a variety of infectious particles such as bacteria and viruses . These superantigens have the capacity to stimulate a large percentage of the host T cells by interacting specifically with the T-cell receptor V beta chain which is shared by about 1-20% of mature T cells . The recent discovery that mammary tumour viruses express such superantigens enabled the analysis of the retroviral life cycle and led to questions about the role of superantigen in amplification of the infection.

Drug Metab Dispos, 1993 Mar-Apr, 21(2), 259 - 67
Microbial models of mammalian metabolism . Furosemide glucoside formation using the fungus Cunninghamella elegans; Hezari M et al.; The diuretic furosemide (Lasix) was metabolized by the fungus Cunninghamella elegans (ATCC 36112) to the phase II conjugate, furosemide acyl glucoside . This metabolite was isolated following semipreparative scale incubations of C . elegans involving glucose nutrient dosing, and was characterized by NMR spectroscopy (1H and 1H/1H correlated), MS (FAB), UV, HPLC with fluorescence detection, and enzymatic treatments . The aglycone fragment of the conjugate was characterized as furosemide by treatment of the metabolite with sodium hydroxide, whereas the sugar part was identified as glucose by cleavage of the conjugate, derivatization of the released sugar, and GC/MS analysis.

J Immunol, 1993 Mar 1, 150(5), 2025 - 32
Priming to heat shock proteins in infants vaccinated against pertussis; Del Giudice G et al.; To investigate whether and in which proportion normal individuals experience a priming to microbial heat shock proteins (hsp), the presence of antibodies to two mycobacterial hsp was tested in serum sample from 2- to 4-mo-old children before and at different times after vaccination with the trivalent vaccine against tetanus, diphtheria, and pertussis (DTP) . We show that 88.9% of infants vaccinated with DTP developed antibody responses to mycobacterial hsp . Such a response was due to the whole-cell pertussis component of the vaccine, because it was not observed in infants receiving an acellular pertussis vaccine . Antibodies and cells reactive to the mycobacterial 65-kDa hsp were also found in mice immunized with DTP . Interestingly, whole-cell pertussis vaccine-induced anti-hsp antibodies cross-reacted with the Escherichia coli GroEL hsp, and at a some extent with the human 60-kDa hsp, belonging to the same hsp family . These data suggest that priming of the immune system to hsp is a common phenomenon occurring very early in life.

J Biotechnol, 1993 Mar, 28(1), 55 - 68
Protein engineering of subtilisins to improve stability in detergent formulations; von der Osten C et al.; Microbial proteases are used extensively in a large number of industrial processes and most importantly in detergent formulations facilitating the removal of proteinaceous stains . Site-directed mutagenesis has been employed in the construction of subtilisin variants with improved storage and oxidation stabilities . It is shown that in spite of significant structural homology between subtilisins subjected to protein engineering the effects of specific mutations can be quite different . Mutations that stabilize one subtilisin may destabilize another.

Phytochemistry, 1993 Mar, 32(4), 799 - 803
Hydrolytic specificity of the barley grain aspartic proteinase; Kervinen J et al.; We recently published the primary structure and inhibition data of the barley grain aspartic proteinase (HvAP, Hordeum vulgare aspartic proteinase) which revealed similarity to mammalian cathepsin D and yeast aspartic proteinase A . Here we present evidence, based on Km and kcat values for the enzyme as well as on its cleavage sites in haemoglobin, the insulin B-chain, glucagon and melittin, that the similarity extends to its hydrolytic specificity . Like the animal and microbial aspartic proteinases, HvAP preferentially cleaves peptide bonds between amino acid residues with large hydrophobic side chains . The narrow hydrolytic specificity of HvAP suggests that plant aspartic proteinases may perform regulatory functions by limited proteolysis.

Appl Environ Microbiol, 1993 Mar, 59(3), 915 - 8
Direct extraction and purification of rRNA for ecological studies; Moran MA et al.; Microgram quantities of rRNA were recovered from natural microbial communities in sediment, soil, and water with a lysozyme-hot phenol direct extraction method . Gel filtration with Sephadex G-75 spun columns readily removed humic-like contaminants without any measurable loss of rRNA and rendered RNA extracts of sufficient quality for molecular procedures.

Lancet, 1993 Feb 27, 341(8844), 515 - 8
High serum procalcitonin concentrations in patients with sepsis and infection; Assicot M et al.; High concentrations of calcitonin-like immunoreactivity have been found in the blood of patients with various extrathyroid diseases . By means of a monoclonal immunoradiometric assay for calcitonin precursors, we have measured serum concentrations of procalcitonin in patients with various bacterial and viral infections . 79 children (newborn to age 12 years) in hospital with suspected infections were investigated prospectively . 19 patients with severe bacterial infections had very high serum concentrations of procalcitonin at diagnosis (range 6-53 ng/mL) in comparison with 21 children found to have no signs of infection (baseline concentrations < 0.1 ng/mL) . Serum procalcitonin values decreased rapidly during antibiotic therapy . 11 patients with peripheral bacterial colonisation or local infections without invasive sepsis and 18 (86%) of 21 patients with viral infections had concentrations within or slightly above the normal range (0.1-1.5 ng/mL) . Among 9 severely burned patients studied in an intensive care unit, the post-traumatic course of procalcitonin concentrations (range 0.1-120 ng/mL) was closely related to infectious complications and acute septic episodes . Concentrations of mature calcitonin were normal in all subjects, whatever procalcitonin concentrations were found . Concentrations of a substance immunologically identical to procalcitonin are raised during septic conditions . Serum concentrations seem to be correlated with the severity of microbial invasion.

Am J Med Genet, 1993 Feb 15, 45(4), 501 - 7
Multiple epiphyseal dysplasia, Fairbank type: morphologic and biochemical study of cartilage; Stanescu R et al.; We have performed histochemical, immunohistochemical, electron microscopic, and biochemical studies on the upper tibial cartilage from a case of multiple epiphyseal dysplasia, Fairbank type . Most chondrocytes had intracytoplasmic inclusions which took the stains for proteins and were resistant to microbial collagenase digestion . The electron microscopic study showed that the inclusions are dilatations of the rough endoplasmic reticulum containing a material with alternately wide electron dense and electron lucent layers . Both in optical and in electron microscopy the inclusions fixed antibodies against the core protein of the large cartilage proteoglycans (aggrecans) . They didn't stain with antibodies against type II collagen . The gel electrophoretic pattern of the large proteoglycans was different from normal controls . The morphologic and biochemical alterations found in multiple epiphyseal dysplasia are similar to those already described in pseudoachondroplasia (Stanescu et al.: Eur J Pediatr 138:121-225, 1982; Stanescu et al.: J Bone Joint Surg 66A:817-836, 1984) . However, the inclusions are smaller and the growth cartilage much less disorganized in multiple epiphyseal dysplasia . The similarity of morphologic and biochemical abnormalities strongly suggests that the two diseases have a similar pathogenesis and belong to the same bone dysplasia family.

Proc Natl Acad Sci U S A, 1993 Feb 15, 90(4), 1267 - 71
T-cell receptor V beta expression in normal human skin; Dunn DA et al.; The skin-associated immune system is the first line of defense against pathogenic attack from the environment and is simultaneously tolerant to localized autoantigens and to antigens of the normal microbial flora . The T-cell receptor (TCR) repertoire of skin lymphocytes may therefore be influenced by the skin microenvironment . We studied the expression of TCR beta-chain variable region (V beta) genes in normal skin by a polymerase chain reaction-based comparative method . When comparing the amplification of V beta genes in peripheral blood and normal skin, we found that TCR V beta 1, -7, -14, and -16 were often highly expressed in skin relative to peripheral blood, whereas V beta 5.1 was often highly expressed in peripheral blood mononuclear cells but not in skin . These results demonstrate that the TCR repertoire of skin lymphocytes is not determined by random sampling of peripheral blood mononuclear cells but may be molded by the interaction with self antigens and/or the normal microbial flora in the microenvironment of the skin.

Zentralbl Bakteriol, 1993 Feb, 278(1), 102 - 11
Complement activation and sensitizing antibodies in Lyme borreliosis . A microbial adherence immobilization assay for Borrelia burgdorferi (MAIA-BB); Cacciapuoti B et al.; A report is presented about the capability of complement to directly clump Borrelia burgdorferi . The new phenomenon which has been termed "microbial adherence", is either antibody-independent or requires the presence of "sensitizing" antibodies depending the strains tested . Microbial adherence is associated with immobilization and killing of borrelias . A microbial adherence immobilization assay for B . burgdorferi (MAIA-BB) was developed to detect sensitizing antibodies in patients with Lyme borreliosis and in B . burgdorferi-infected animals.

Acta Paediatr, 1993 Feb, 82(2), 137 - 41
Antigen and antibody assays in the aetiological diagnosis of respiratory infection in children; Korppi M et al.; The diagnostic efficacy of two methods--demonstration of seroconversion in paired sera and detection of antigen in clinical specimens--was evaluated in 183 children with respiratory syncytial, parainfluenza or adenoviral, or pneumococcal respiratory tract infection . Viral infection was diagnosed in 46 (37%) of the 125 cases by antigen assay alone, in 36 (29%) by antibody assay alone and in 43 (34%) by both methods . In respiratory syncytial viral infections, antigen assays were more often positive than antibody assays; 80% of the cases were antigen positive and 63% solely antigen positive . In parainfluenza and adenoviral infections, antigen assays were not as useful; a positive result was seen in 59% and 44% of cases, respectively . Pneumococcal infection was diagnosed in 25 (30%) of the 84 cases by antigen assay alone, in 54 (64%) by antibody assay alone and in only 5 (6%) by both methods . Thus nearly all pneumococcal infections were diagnosed by only one method . The efficacy of antigen and antibody assay was clearly dependent on the age of the patients . In infants less than six months of age, nearly all (27 (90%)) of the 30 infections were diagnosed by antigen detection; 25 solely by antigen detection . In older patients, antigen and antibody assays supplemented each other . We conclude that antigen detection should be used as the primary method for the diagnosis of viral or pneumococcal respiratory tract infection . In infants, antigen detection is the only reliable method of microbial diagnosis . In addition, antigen detection is the method of rapid microbial diagnosis . In pneumococcal infections, the sensitivities of antigen detection methods are not sufficient; this is an important area for further research.

Zentralbl Hyg Umweltmed, 1993 Feb, 193(5), 395 - 418
{Environmental medicine group diagnosis of children--studies from 1982 to 1990 . Review}; Beck EG et al.; In the period of 1982-1990 a total of 5760 children (2893 boys, 2816 girls) were examined in the frame-work of the Clean Air Programme Rhine-Main: from the polluted Rhine-Main area 2511 children (Wiesbaden 1682, Frankfurt since 1985, 825), from the control areas 3153 children (Freiburg area 1937, Starnberg area 1316) . Healthy children living in the industrialised Rhine-Main area exhibited alterations of the following parameters compared with healthy children living in the less polluted control areas: Long-term alterations, enlarged and fissured, rough palatine tonsils, palpably enlarged cervical and mandibular lymph nodes, Short-term alterations, erythrocyte count: from 1981-1988 relatively smaller numbers, from 1988-1990 equalized to the scatter range of controls, hemoglobin values: corresponding to erythrocyte counts, activity of lymphocyte nucleoli: increased until 1985, later on equalized, lymphocyte count: reduced, T8-suppressor cell count: decreased, chemiluminescence of granulocytes: increased, palatine tonsils: altered microbial contamination with contact germs, altered physiological resident flora, heavy metals (Pb, Cd) in hair: from 1982-1990 increased, then slowly reduced corresponding to the results of the ambient air analyses . These findings indicate an increased inclination for reaction of the healthy juvenile organism in the sense of biological adaptation to long-term stress, i.e . living for years in an area with air pollution, and are also in good agreement with the measuring values characterising the ambient air quality.

Protein Eng, 1993 Feb, 6(2), 213 - 9
Expression of functional papain precursor in Saccharomyces cerevisiae: rapid screening of mutants; Vernet T et al.; A microbial expression system for the study of the cysteine protease papain has been developed as a more useful alternative to the insect cell/baculovirus expression system we have previously used . A synthetic papain precursor (propapain) gene was expressed in the yeast Saccharomyces cerevisiae under the control of the alpha-factor promoter . Efficient expression required fusion of the propapain sequence with the yeast alpha-factor prepro region and a yeast host cell defective in the synthesis of vacuolar proteases . Surprisingly, the glycosylated form of the inactive papain precursor is not secreted, but accumulates within the yeast cell . Complete conversion of the intracellular zymogen into active mature papain could be achieved in vitro . Purified recombinant papain produced by the yeast system has kinetic characteristics similar to those of the natural enzyme . An advantage of the yeast expression system over the baculovirus/insect cell system is that we can perform mutagenesis and screening of papain mutants very efficiently . We have set up a 'one-tube' screening procedure for the simultaneous characterization of numerous mutants of the papain precursor . Yeast cells are grown and lysed in microtiter plate wells and the released papain precursor is then activated to mature papain . This assay allows easy discrimination between proteins with close to wild type properties and proteins that are not functional . We have applied this assay to investigate the spectrum of amino acids which are tolerated at Asn175 of papain using two independently derived libraries of mutants at this position . Many amino acid substitutions at this position are not accepted; only the reintroduction of Asn restored normal function.

Biol Chem Hoppe Seyler, 1993 Feb, 374(2), 101 - 9
Microbial metabolism of quinoline and related compounds . XVI . Quinaldine oxidoreductase from Arthrobacter spec . Rü 61a: a molybdenum-containing enzyme catalysing the hydroxylation at C-4 of the heterocycle; de Beyer A et al.; Quinaldine oxidoreductase from Arthrobacter spec . Ru 61a converts quinaldine to 1H-4-oxoquinaldine . The enzyme was purified 70-fold to apparent homogeneity in a 5-step procedure with a recovery of 4% . The molecular mass of the native enzyme was calculated to be 340,000 Da by gel filtration . SDS-polyacrylamide gel electrophoresis of the enzyme revealed 3 protein bands corresponding to 82,000 Da, 35,000 Da and 22,000 Da . The enzyme contained 1.6 atoms of molybdenum, 8 atoms of iron, 8 atoms of acid labile sulfur, 2 molecules of FAD and as part of the molybdenum cofactor, molybdopterin cytosine dinucleotide . Due to the composition of the cofactors the quinaldine oxidoreductase belongs to the class of molybdo-iron/sulfur-flavoproteins . Cyanide, arsenite and 4-hydroxymercuribenzoate were effective inhibitors whereas the enzyme was not affected by methanol.

Diagn Microbiol Infect Dis, 1993 Feb, 16(2), 153 - 7
Molecular technology for hospital epidemiology; Miller JM; Hospital-acquired infections in the United States contribute to approximately 80,000 deaths per year, with an associated cost of > $4 billion . Some of these infections are associated with outbreaks and clusters occurring within the hospital . The hospital infection control team must respond quickly and decisively to recognize and curtail these outbreaks, but their response often depends on critical laboratory data that characterize or "fingerprint" suspected microbial isolates . This presentation summarizes the simple, easily performed tests and the more molecular approaches that a hospital laboratory may take in support of infection control efforts . In addition to phenotypic data, such as biotype and antibiograms, hospitals that elect to incorporate molecular protocols should limit their first experiences to plasmid analysis and plasmid restriction endonuclease profiles.

Lab Anim Sci, 1993 Feb, 43(1), 78 - 85
Kong toys for laboratory primates: are they really an enrichment or just fomites?
Bayne KA, Dexter SL, Hurst JK, Strange GM, Hill EE.
Simple toys as enrichment devices have been associated with a rapid decline in their use by nonhuman primates . Other facets of toy presentation have not been described previously . For example, a comparison of the effect(s) of an enrichment device between two facilities should be validated if enrichment recommendations are to be made that affect diverse research facilities across the country . Additionally, a comparison of two methods of presentation (one highly accessible to the animal and the other less accessible) of the same enrichment device for potential differences in efficacy could provide direction in implementing an enrichment program based on simple toys . The handling of enrichment devices by nonhuman primates can lead to the spread of microbial contamination . The typical enrichment program rotates enrichment devices among animals to maximize the variety of stimuli available to each primate in the most economic manner . An adequate sanitation program is therefore pivotal to minimizing the potential for enrichment devices to be fomites . We conducted three experiments that addressed these issues . The results confirmed that, although the presence of a simple toy reduced behavioral pathology, there was variability in behavioral effect for an enrichment technique between facilities . Two methods of presentation (on floor and suspended) of a simple toy did not produce any significant differences in use . Finally, we demonstrated that microbial growth can persist on enrichment devices after they have been sanitized in a commercial cagewasher.

Carcinogenesis, 1993 Feb, 14(2), 205 - 9
Inhibition of tumor formation from grafted murine papilloma cells by treatment of grafts with staurosporine, an inducer of squamous differentiation; Strickland JE et al.; The microbial alkaloid staurosporine induces responses associated with protein kinase C activation, resulting in terminal differentiation in cultures of both normal and neoplastic mouse epidermal cells . As a cancer chemotherapy model, we treated grafts of mouse epidermal tumor cell lines 308 and SP-1 repeatedly with staurosporine . A dose-dependent inhibition of tumor formation, maximal at 0.025 nmol per treatment, was observed . Higher and lower doses were less effective, suggesting a specific target for staurosporine action . A single, low-dose treatment 2 weeks after grafting also markedly reduced tumor formation . Although in vitro evidence suggests that staurosporine-induced terminal squamous differentiation results from activation of protein kinase C, we found no inhibition of tumor growth in similar studies with the protein kinase C activator 12-O-tetradecanoylphorbol-13-acetate . These results indicate that staurosporine is an effective antitumor agent for eradicating squamous cell tumors in vivo.

Bone Marrow Transplant, 1993 Feb, 11(2), 147 - 54
A novel approach to immunomodulation of frozen human bone marrow with interleukin-2 for clinical application; Charak BS et al.; Interleukin-2 (IL-2) activation of fresh or frozen bone marrow (BM) in vitro generates killer cells with potent anti-tumor effect both in vitro and in vivo . The IL-2-activated BM (ABM) retains the capacity to reconstitute the hematopoietic system in an autologous bone marrow transplantation (ABMT) setting . The killer cells lose their cytotoxicity if the ABM undergoes the procedures of freezing and thawing . Therefore, for clinical application, the ABM has to be generated after thawing a frozen stock of BM before ABMT . The thawed BM cells are fragile and may undergo lysis, resulting in clump formation and cell loss . The frozen autograft also contains components of cryoprotectant mixture whose effects on the generation of ABM have not been defined . The present studies have been carried out to optimize a technique of handling the frozen BM for immunomodulation with IL-2 for 24 h at 37 degrees C prior to ABMT, with minimal loss of cells . IL-2-activation of BM was carried out in bags containing serum free medium which were designed to permit gaseous exchange . Addition of deoxyribonuclease (DNAse) (100 micrograms/ml of BM concentrate) immediately after thawing and the presence of heparin (20 units/ml) in the medium completely abrogated immediate or delayed clumping of cells . The presence of DNAse and/or heparin during in vitro culture did not affect the cell viability, cytotoxicity against tumor cells or the progenitor cell activity of the ABM; all these functions were well maintained even when BM was placed in culture immediately after thawing (without washing) . There was no microbial contamination.(ABSTRACT TRUNCATED AT 250 WORDS)

Appl Environ Microbiol, 1993 Feb, 59(2), 410 - 6
Effect of phenotypic plasticity on epiphytic survival and colonization by Pseudomonas syringae; Wilson M et al.; The bacterial epiphyte Pseudomonas syringae MF714R was cultured on agar or in broth or collected from colonized leaves; it was then inoculated onto greenhouse-grown bean plants incubated in a growth chamber at low relative humidity or in the field or onto field-grown bean plants . Cells cultured in liquid medium survived the least well after inoculation of leaf surfaces under all conditions . Cells cultured in solid medium exhibited the highest percent survival and desiccation tolerance in the growth chamber but generally survived less well in the field than did cells harvested from plants . Cells harvested from plants and inoculated onto plants in the field usually exhibited the highest percent survival, started to increase in population earlier, and reached a higher number than did cells cultured in vitro . Differences in field survival were apparently not attributable to differential UV tolerance . The observed effects of phenotypic plasticity on epiphytic survival and colonization should be considered in risk assessment studies, in studies of bacterial epidemiology, and in the use of microbial antagonists for biological pest control.

J Clin Invest, 1993 Feb, 91(2), 717 - 23
Major cytoplasmic membrane protein of Legionella pneumophila, a genus common antigen and member of the hsp 60 family of heat shock proteins, induces protective immunity in a guinea pig model of Legionnaires' disease; Blander SJ et al.; We have examined the capacity of the major cytoplasmic membrane protein (MCMP) of Legionella pneumophila, a genus common antigen and member of the hsp 60 family of heat shock proteins, to induce protective immunity in a guinea pig model of Legionnaires' disease . We purified MCMP to homogeneity from L . pneumophila by buffer extraction, ion-exchange chromatography, and molecular sieve chromatography . Guinea pigs immunized with MCMP developed a strong cell-mediated immune response to the immunogen manifest by marked cutaneous delayed-type hypersensitivity . Guinea pigs immunized with MCMP and then challenged with a lethal aerosol dose of L . pneumophila exhibited a high level of protective immunity . Altogether, in four independent experiments, 55 of 64 (86%) animals immunized three times with 0.6-40 micrograms MCMP including 11 of 11 (100%) animals immunized three times with 40 micrograms MCMP survived aerosol challenge with L . pneumophila compared with 1 of 29 (3%) sham-immunized control animals (P < 0.0001, Cochran-Mantel-Haenszel X2 statistic for pooled data) . To our knowledge, MCMP is the first member of the hsp 60 family of proteins shown to induce protective immunity to a microbial pathogen . MCMP has potential as a vaccine against Legionnaires' disease . Since MCMP is a genus common antigen, vaccination with a combination of MCMPs derived from different Legionella species has the potential of inducing protective immunity against all the major Legionella species causing human disease.

J Environ Sci Health B, 1993 Feb, 28(1), 67 - 80
Effect of fungicides, captafol and chlorothalonil, on microbial and enzymatic activities in mineral soil; Tu CM; Tests were conducted to determine the effects of fungicides, captafol and chlorothalonil, on microbial and enzymatic activities in sandy loam . The results indicated that when captafol or chlorothalonil was added to the sandy loam, bacterial and fungicidal populations initially decreased with the treatments but recovered rapidly to levels similar to those in the controls . No inhibition on oxidation of soil ammonia or organic sulfur was observed . The fungicide treatments significantly increased oxygen consumption from the decomposition of organic matter indigenous to the soil . Both fungicides suppressed invertase and amylase for 1 day . However, the inhibitory effect disappeared after 2 days . Captafol depressed dehydrogenase for 4 days and recovered to equal to that of control after 7 days . No inhibitory effect on urease and phosphatase was shown with the fungicidal treatments . Although some stimulatory influences of fungicides on microbial and enzymatic activities were found in the soil, in no instance were the effects dramatic or sufficient enough to be considered important to soil fertility.

J Anim Sci, 1993 Feb, 71(2), 499 - 504
Organic matter and nitrogen digestion by dairy cows fed calcium salts of rapeseed oil fatty acids or rapeseed oil; Doreau M et al.; The aim of this experiment was to study the effects on digestion of a supply of Ca salts of vegetable oil as a fat source for ruminants . Total tract digestibility and ruminal digestion were studied using three ruminally and duodenally fistulated dairy cows in a Latin square design . Diets compared were a control diet (C) based on corn silage and concentrate, and the C diet supplemented with calcium salts of rapeseed oil (S) or supplemented with rapeseed oil (O) . Organic matter digestibility was lower (P < .05) for diet O than for diet C; diet S was intermediate and not different (P > .05) from diet C . Treatment did not affect (P > .05) the proportion of OM digested in the rumen, total N and microbial N flow at the duodenum, or efficiency of N microbial synthesis . In situ degradation of DM and NDF was higher for diet S than for diet C . The molar concentration of ruminal acetate was lower (P < .05) for O than for C and S . Under the conditions of this trial, supplemental calcium salts rich in polyunsaturated fatty acids did not decrease ruminal digestion.

Rheum Dis Clin North Am, 1993 Feb, 19(1), 207 - 22
Microbial superantigens as etiopathogenic agents in autoimmunity; Friedman SM et al.; A recently characterized group of immunologically active microbial products, termed superantigens, may provide the etiopathogenic link between antecedent infection and the subsequent development of autoimmunity in the genetically susceptible host . In this article, the authors review the cellular interactions that underlie autoimmune disease and emphasize the central role played by T lymphocytes . Based on the unique properties of the superantigen and data derived from experimental animal models and clinical studies of human autoimmune disease, a hypothesis that these molecules could trigger both systemic and organ-specific autoimmunity was developed.

J Ind Microbiol, 1993 Feb, 12(2), 76 - 86
Whole cell biocatalysis in nonconventional media; Nikolova P et al.; In this paper biocatalytic reactions carried out by whole cells in nonconventional media are reviewed . Similar relationships are observed between solvent hydrophobicity and catalytic activity in reactions carried out by isolated enzymes and whole cells . In addition to the effect of organic solvent on biocatalyst stability, microbial cells are susceptible to damaging effects caused by the organic phase . In general, more hydrophobic solvents manifest lower toxicity towards the cells . Whole cell biocatalysts require more water than isolated enzymes and two-phase systems have been most widely used to study whole cell biocatalysis . Immobilization makes cell biocatalysts more resistant to organic solvents and helps achieve homogeneous biocatalyst dispersion . Cell entrapment methods have been widely used with organic solvent systems and mixtures of natural and/or synthetic polymers allow adjustment of the hydrophobicity-hydrophilicity balance of the support matrix . Some examples of stereoselective catalysis using microbial cells in organic solvent media are presented.

J Antibiot (Tokyo), 1993 Feb, 46(2), 214 - 21
Anthrotainin, an inhibitor of substance P binding produced by Gliocladium catenulatum; Wong SM et al.; Substance P (SP) is an undecapeptide belonging to a family of chemically related neurotransmitters and neuromodulators known as neurokinins . In our search for SP antagonists, we screened microbial broth extracts for the ability to inhibit radiolabeled SP binding to membranes prepared from rat forebrain . Anthrotainin was isolated from a fungal culture and determined to be a novel tetracyclic compound, with an IC50 of 3 microM against {125I}SP . The structure, spectroscopic, chemical, and pharmacological properties of anthrotainin are presented.

Diagn Microbiol Infect Dis, 1993 Feb, 16(2), 159 - 63
Diagnostic molecular microbiology . Current challenges and future directions; Persing DH; The advent of nucleic acid amplification techniques for the clinical laboratory provides not only new diagnostic opportunities but new responsibilities as well . Problems associated with the introduction of this technology include contamination with the products of amplification reactions, and difficulty in interpreting test results . Eventually, however, many of these problems will be overcome, and new applications of diagnostic molecular microbiology such as sequence-based microbial identification will become established . Clinical microbiologists will successfully negotiate the transition to tests based on nucleic acid chemistry if they are willing to educate and become educated in the new technology.

J Gen Microbiol, 1993 Feb, 139 ( Pt 2), 371 - 8
A new polymorphic methanogen, closely related to Methanocorpusculum parvum, living in stable symbiosis within the anaerobic ciliate Trimyema sp; Finlay BJ et al.; A new anaerobic microbial consortium has been discovered: the partners are the ciliated protozoon Trimyema sp . and a single species of methanogen . The consortium has been maintained in culture for more than four years . Each ciliate contains up to 300 symbiotic bacteria; many are relatively small and irregularly disc-shaped, and these are distributed throughout the host's cytoplasm, whereas those which are attached to the ciliate's hydrogenosomes are significantly larger and profusely dentate . This attachment is interpreted as an adaptation to maximize capture by the bacteria of the H2 escaping from hydrogenosomes . The 16S rRNA gene of the symbionts has been partially sequenced, and fluorescent oligonucleotide probes have been constructed and used to detect the different morphotypes of the symbiont within the ciliate . The symbionts belong to a new species of archaeobacterium which is a close relative of the free-living methanogen Methanocorpusculum parvum.

Biochem J, 1993 Jan 15, 289 ( Pt 2), 417 - 22
Purification and characterization of the major glutathione transferase from adult toad (Bufo bufo) liver; Aceto A et al.; Five forms of glutathione transferase (GST) were resolved from the cytosol of adult common toad (Bufo bufo) liver by GSH-affinity chromatography followed by isoelectric focusing . The major enzyme (GST-7.64; 55% of total activity bound to the column) has a pI value of 7.64, is composed of two subunits each with a molecular mass of 23 kDa, and has the N-terminal amino acid residue blocked . GST-7.64 has also been characterized with respect to amino acid composition, substrate specificity, inhibition characteristics, c.d . spectra and immunological reactivity . The N-terminal sequence of some peptides obtained after tryptic digestion has also been determined . All together the results obtained suggest that the major toad liver GST is distinct from any known GST, including microbial, plant and mammalian GSTs.

Science, 1993 Jan 8, 259(5092), 224 - 7
Presentation of a viral T cell epitope expressed in the CDR3 region of a self immunoglobulin molecule; Zaghouani H et al.; Synthetic peptides corresponding to microbial epitopes stimulate T cell immunity but their immunogenicity is poor and their half-lives are short . A viral epitope inserted into the complementarity-determining region 3 (CDR3) loop of the heavy chain of a self immunoglobulin (Ig) molecule was generated from the Ig context and was presented by I-Ed class II molecules to virus-specific, CD4+ T cells . Chimeric Ig-peptide was presented 100 to 1000 times more efficiently than free synthetic peptide and was able to prime virus-specific T cells in vivo . These features suggest that antigenized Ig can provide an improved and safe vaccine for the presentation of microbial and other peptides.

Arch Tierernahr, 1993, 45(3), 263 - 79
Digestion characteristics by beef cattle consuming bermudagrass or bromegrass hay alone or with alfalfa and(or) corn; Sun W et al.; Cannulated beef cattle (four cows: 556 kg initial weight; four steers: 504 kg initial weight) were used in an experiment with two simultaneous Latin squares to determine effects of substituting alfalfa and(or) corn for vegetative bermudagrass (BER; 77% neutral detergent fibre and 5.5% acid detergent lignin) or mature bromegrass (BRO; 70% neutral detergent fibre and 6.6% acid detergent lignin) hay on digestion characteristics . For Controls, BER or BRO was fed at 1.32 or 1.54% body weight of cows and steers, respectively; other treatments entailed substitution for hay DM of alfalfa cubes (17%) or ground corn (33%) . A protein supplement was given to all animals . In vitro neutral detergent fibre (NDF) digestion was slightly greater for BER than BRO . Supplement treatments did not affect the concentration of total volatile fatty acids or the molar proportion of propionate in ruminal fluid . True ruminal organic matter (OM) digestion was similar among diets; greater duodenal microbial OM flow and postruminal NDF digestion for BRO than BER diets were responsible for higher (P < 0.05) postruminal OM digestion for BRO diets . Supplement treatment did not affect duodenal microbial nitrogen flow or efficiency of microbial growth . Corn supplementation increased total tract OM digestion (P < 0.05) . Alfalfa addition depressed total tract OM and NDF digestibilities more when added to BER than BRO; depressions in total tract NDF digestion with alfalfa and corn substitutions were additive . With constant DM intake, slightly less than ad libitum, alfalfa or corn substituted alone or together for hay did not improve characteristics of digestion by cattle consuming vegetative bermudagrass or mature bromegrass other than increased postruminal and total tract OM digestibilities with corn.

Arch Tierernahr, 1993, 45(3), 235 - 44
{Use of microbial phytase at different phosphorus supply levels in broiler fattening . 1 . Effect on fattening performance and tibia stability}; Richter G; Two trials with 720 male and female Lohmann-broilers each and a fattening period of 35 days were carried out to study the effects of different phytase supplementations (0; 300; 700 units per kg feed) . The basal diets with only ingredients of plant origin or of plant and animal origin contained between 0.4 and 0.5 per cent total-P . The supplementation of inorganic P was 0; 338; 675 and 1350 mg per kg feed . The feed intake increased on average by 15 and 17% resp . after adding of 300 or . 700 units phytase to diets with different phosphorus supply . The body weight of broilers increased also in dependence of feed intake . The feed conversion was not influenced by the different phytase and inorganic P-supply . The bone stability examined as bending and breaking strength of tibia was improved by supplementation of phytase and inorganic P . Based on the fattening parameters 300 units phytase can replace 0.06 per cent inorganic P in the diet.

Caries Res, 1993, 27(3), 169 - 75
An in vitro study into the effect of a bacterial artificial caries system on the enamel adjacent to composite and amalgam restorations; Gilmour AS et al.; A microbial artificial caries system was used to produce caries-like lesions adjacent to amalgam and composite restorations . Polarised microscopy allowed measurement of outer and wall lesions depths . Comparison was made of the depths found in relation to the restorative materials, margin finishing procedures (composite group), varnish application (amalgam group) and thermal stressing . It was found that differences were detectable between the different restorative materials and the type of margin finishing procedure used in the composite group . Varnish application and thermal stressing had little effect . Some comparison is made between this microbial technique, other artificial caries techniques, and other methods of assessing marginal leakage.

Scand J Infect Dis, 1993, 25(2), 207 - 13
C-reactive protein in viral and bacterial respiratory infection in children; Korppi M et al.; C-reactive protein (CRP) was studied in 209 children treated in hospital due to middle or lower respiratory tract infection with serologically demonstrated viral or bacterial aetiology . Of the 110 patients with serological evidence of bacterial infection, either alone or in association with viral infection, 52% had CRP > 20 mg/l, 35% > 40 mg/l and 15% > 80 mg/l . Of the 99 patients with serological evidence of viral infection alone, 35% had CRP > 20 mg/l, but only 12% > 40 mg/l and 5% > 80 mg/l . Nearly all, 88%, of the 25 patients with CRP > 40 mg/l in association with viral infection had either an infectious focus, specific microbial or non-specific laboratory evidence suggestive of bacterial infection . By calculating diagnostic parameters at 3 cut-off levels of CRP, the level 40 mg/l seemed more useful than 20 mg/l or 80 mg/l for differentiation between viral and bacterial infections . By using a CRP value of 40 mg/l as a screening limit sensitivity was 0.55, specificity 0.88, positive predictive value 0.76, negative predictive value 0.55, and likelihood ratios of a positive and negative test result 2.9 and 0.74, respectively . It is concluded that low CRP values do not rule out bacterial aetiology of respiratory infection in children . On the other hand viral infection without bacterial involvement is very improbable if CRP is > 40 mg/l . Our results suggest that high CRP values rule out viral infection as a sole aetiology of infection; bacterial infection and antibiotic treatment should be considered in these cases.

Vet Med (Praha), 1993, 38(2), 83 - 8
{A rapid method of detecting psychrotrophic bacteria in raw milk}; Urbanova E; Determination of the counts of psychrotrophic bacteria offers an important indicator of the technological quality of purchased raw milk, and also an indicator of the hygienic level of its production in agricultural primary production, particularly with respect to the sanitation of milking machines . The applied and published methods of determination of the counts of psychrotrophic microbial contaminants have revealed large differences, multiplied by insufficient legislation in this country, by vague and even wrong definition of the given group of bacteria, and/or by the misuse of the concepts psychrotrophic and psychrophil bacteria . Since the level of the microbial quality of raw milk should be increased, we investigated and developed convenient methods of determination of psychrotrophic bacterium counts in the last years (Urbanova et al., 1988; Urbanova, in print): these methods should facilitate the reasonable control applying an expeditious procedure . A long incubation time of standard procedure is a disadvantage complicating the practical use of the previous procedure (10 days at a temperature of 7 degrees C) . An accelerated procedure of psychotrophic bacterium cultivation to determine their counts in raw milk is described in this paper . After thorough stirring, a raw milk sample is diluted with sterile physiological solution (pH 7.2) pursuant to the standard CSN 57 0101 . The dilutions 10(-2), 10(-3), 10(-4) were the most convenient for inoculation when the counts of psychrotrophic contaminants were determined in raw milk . The dilutions 10(-3), 10(-4), 10(-5) were inoculated if higher milk contaminations were expected.(ABSTRACT TRUNCATED AT 250 WORDS)

Antonie Van Leeuwenhoek, 1993 Jan, 63(1), 39 - 43
Why it pays for bacteria to delete disused DNA and to maintain megaplasmids; Stouthamer AH et al.; Genetic information for disused metabolic systems is easily lost . This can be understood on the basis of a mathematic model for the description of the growth of individual microbial cells . The essential features of the model are that the uptake of nutrients is proportional to the surface area of the cell and maintenance costs are proportional to the volume . This explains why disused genes are easily lost . Growth of the individual cell continues until DNA replication of the genome is completed . The consequence is that cells with a large genome size are bigger at the moment of cell division . This results in a less favourable surface to volume ratio, which has a negative influence on the population growth rate . This means that cells with a smaller genome size will have a selective advantage . In cells in which the total DNA is divided over several units which replicate simultaneously (e.g . a cell with more chromosomes or a cell with one chromosome and one or more megaplasmids) a high surface to volume ratio can be maintained . The great metabolic differences between phylogenetically strongly related bacteria are due to loss of genetic information for complex metabolic characters in order to maintain a small genome size and a favourable surface to volume ratio.

Agents Actions, 1993 Jan, 38(1-2), 85 - 90
Macrolide antibiotics as antiinflammatory agents: roxithromycin in an unexpected role; Agen C et al.; The antiinflammatory activity of a new 14-membered macrolide antibiotic, roxithromycin, was evaluated in various rat models including carrageenan- and poly-L-arginine-induced hind-paw oedema, croton oil inflamed ear assay and polyester sponge granuloma . When administered orally to animals, roxithromycin displayed an atypical profile in the assays utilized, including: (1) marked antioedema activity similar to that of indomethacin in poly-L-arginine assay, (2) significant inhibition of lambda-carrageenan hind-paw oedema and croton-oil-induced inflammation in the ear, although indomethacin was more effective, and (3) failure to reduce the development of granuloma induced by implanted polyester sponges, while indomethacin significantly reduced the chronic inflammatory reaction . Based on these results, it is concluded that roxithromycin is active in reducing the acute inflammatory reaction in rat models through mechanisms different from conventional nonsteroidal antiinflammatory agents such as indomethacin . Therefore, roxithromycin may have a favorable impact on skin inflammatory reactions accompanying microbial infections.

Respiration, 1993, 60 Suppl 1, 16 - 24
Role of the immune system in recovery from infection; Konig W; Infection is regarded as an imbalance between microbial pathogenicity factors and the host defense system . The virulence mechanisms include adhesion, chemotaxis, invasion, resistance and production of toxins . In addition, local and/or systemic immune functions in patients undergoing infections are altered . Nonspecific as well as specific cellular and humoral defense mechanisms are affected . The interaction of defined microbial pathogenicity factors with immune effector cells results in the activation of a variety of inflammatory mediators; they are a prerequisite for protective immunity but also induce local or systemic damage in the host when they occur in excessive amounts and when their metabolism is inadequately controlled . The analysis of the pathophysiological events during infection in patients by taking advantage of modern molecular and cell-biological methods may contribute to the development of novel therapeutic strategies.

J Basic Microbiol, 1993, 33(1), 19 - 25
Microbial degradation of aquatic biomass by Trichoderma viride 992 and Aspergillus wentii 669 with reference to the physical structure; Kour J et al.; There was a considerable change in the structural parameters of both the lignocellulosic substrates . This change was confirmed by infrared (IR) spectroscopy and X-ray diffraction data, which revealed the presence of phenolic groups in the peaks range 1310-1410 and 1200 cm-1 with large crystalline substrates . These were found to be reduced after treatment with alkali . Enzyme/s activity were found to be increased after pretreatment of hydrolysed substrates . The structural modifications in both the substrates were further evidenced by the results obtained after enzymatic hydrolysis of alkali-treated substrates.

Prikl Biokhim Mikrobiol, 1993 Jan-Feb, 29(1), 70 - 9
{Propagation of toxigenic fungi and mycotoxins in various grains}; L'vova LS et al.; The frequency of occurrence and toxigenicity of various strains of Aspergillus flavus (663 stairs) and of Fusarium spp . (459 strains) isolated from grain of various crops from four grain-producing areas were studied . The analysis of mycotoxins in 1038 grain samples (wheat, rye, rice and corn) and in 58 samples of sun flower seeds collected from commercial batches showed that aflatoxins B1 and G1, deoxynivalenol, and zearalenone are the basic microbial contaminants of grain, while toxin T-2 and diacetoxyscirpenol were found much more rarely . Ochratoxin A, penicillic and kojic acids were found only in several samples . The occurrence of the mycotoxins and contamination levels depended on grain-producing areas . The risk of mycotoxins invasion in grains was greater in southern humid regions of Russia, Kazakhstan and Uzbekistan.

Prikl Biokhim Mikrobiol, 1993 Jan-Feb, 29(1), 155 - 60
{Characteristics of lipids of microbial origin with regard to triglyceride composition and comparison of them to plant oils}; Liapkov BG et al.; The triacylglycerol composition of lipids isolated from the yeasts Candida guilliermondii and C . humicola and of food vegetable oils was investigated by HPLC . The triacylglycerol composition of samples was compared using the program "Nearest neighbour" and by plotting images with the help of the software package STATGRAPHICS . The methods used allow us to conclude that microbial lipids are rather promising.

J Anim Sci, 1993 Jan, 71(1), 3 - 10
Influence of processing on the comparative feeding value of barley for feedlot cattle; Zinn RA; Ninety-six crossbred steers were used in a 172-d feedlot growth performance trial to determine the effects of type processing on the comparative feeding value of barley in a 90% concentrate finishing diet . Treatments were: 1) steam-flaked corn (SFC; density = .31 kg/L); 2) dry-rolled barley (DRB; density = .39 kg/L); 3) steam-rolled barley, coarse roll (SRB-C, density = .39 kg/L); and 4) SRB, thin roll (SRB-T, density = .19 kg/L) . The ADG was similar (P > .10) across barley treatments, averaging 1.29 kg/d . Feed intake (P < .05) was lower for SRB than for DRB . Diet NE was greater (P < .05) for SRB than for DRB . Feed intake was lower (P < .05) for SFC than for barley diets . The influence of grain processing on characteristics of digestion was evaluated using four Holstein steers (average BW = 230 kg) with cannulas in the rumen and proximal duodenum . Ruminal and total tract digestibility of OM and starch were lower (P < .05) for DRB than for SRB . Ruminal and total tract digestibility of OM and starch were similar (P > .10) for SRB-C and SRB-T . However, passage of nonammonia N was markedly increased (26.9%, P < .01) with SRB-T compared with SRB-C . This increase in ruminal N efficiency was due partly to reduced ruminal degradation of feed N (23.4%, P < .01) and partly to increased microbial N synthesis (13.3%, P > .10) . The comparative feeding value of DRB, SRB-C, and SRB-T in this study was 90, 92, and 96% the value of SFC, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Zentralbl Mikrobiol, 1993 Jan, 148(1), 16 - 23
Microbial biomass-persistence relationships of acifluorfen in a clay-loam soil; Perucci P et al.; The interference of the effect of the herbicide acifluorfen on microbial biomass and on hydrolytic capacity, and its persistence in a clay-loam soil before and after enrichment with glucose, were investigated . The experiment was carried out under laboratory conditions for 120 days . Acifluorfen was added to the soil at two different application rates corresponding to 1X and 10X the recommended field rate . Biomass-C was significantly higher in the enriched soil during the first 35 days; subsequently there was a tendency to return to the original value of the unenriched soil . The herbicidal treatments depressed the biomass-C level, particularly at the higher rate . The hydrolytic capacity, measured as FDA-hydrolase activity, was significantly higher in the enriched soil than in the unenriched soil . This was enhanced by acifluorfen treatment, chiefly at the higher rate . The degradation trend of acifluorfen was not significantly different at the two rates, but was significantly faster in the enriched soil . Half-life values of 28 and 40 days were found in the enriched and unenriched soil, respectively.

Mikrobiol Zh, 1993 Jan-Feb, 55(1), 85 - 8
{A method for determining the content of anionogenic surface-active peptidolipids of bacterial origin}; Shul'ga AN et al.; The simple and selective method of anionic biosurfactant concentration determination in bacterial cultural broth is offered . The method is based on the ability of the anionic surfactants to form some coloured complex with the cationic indicator methylene blue registered by spectrophotometer . The suggested method is recommended for the study of biosynthesis of microbial surfactants.

J Parenter Sci Technol, 1993 Jan-Feb, 47(1), 9 - 15
Validation and control of non-standard sterilization processes . Committee on Microbial Purity . FIP (International Pharmaceutical Federation); Dipeptidyl peptidase I from goat brain: purification et al.; Department of Chemistry, Kurukshetra University, Haryana, IndiaBrain dipeptidyl peptidase (DPP) I has been purified 2990-fold to apparent homogeneity shown by a single protein band in electrophoreses at pH 4.5, 8.4 and in SDS-PAGE at pH 7.2 . The purification techniques included homogenization of brain acetone powder, autolysis at pH 4.2, 30-80% (NH4)2SO4 fractionation . Sephadex G-100 column chromatography, heat treatment at 65 C . organomercurial affinity chromatography . CM-Sephadex cation-exchange chromatographies at pH 5.6 and 5.0 and anion-exchange chromatography on DEAE-Sephadex at pH 6.8 . The enzyme hydrolysed synthetic substrate Gly-Arg-4-methoxy-beta-naphthyl-amide maximally at pH 6.0 . The Km values for Gly-Arg-beta-naphthylamide and Gly-Arg-4-methoxy-beta-naphthylamide substrates were 0.10 mM and 0.14 mM respectively . The enzyme was inhibited by thiol inhibitors like p-chloromercuribenzoic acid, iodoacetic acid, iodoacetamide and microbial inhibitors leupeptin and antipain . Molecular weight estimations on a calibrated Sephadex G-200 column afforded a value of 180,000 Da while in denaturing conditions on sodium dodecyl sulphate polyacrylamide gel electrophoresis, the subunit molecular weight was 22,000 Da . The subunit structure of the native enzyme was unfolded in presence of different concentrations of urea . In 8 M urea, the enzyme dissociated completely into monomers of 25,000 Da but 6, 5 and 4 M urea concentrations revealed the existence of dimers, tetramers and hexamers . Leu-enkephalin . Tyr-Gly-Gly-Phe-Leu was degraded by DPPI into Tyr-Gly and Gly-Phe-Leu with no further degradation of the newly generated tripeptide.

Arch Microbiol, 1993, 159(2), 119 - 23
Effect of growth temperature on the fatty acid composition of Mycobacterium phlei; Suutari M et al.; In Mycobacterium phlei, fatty acid unsaturation increased with decreasing temperature . The 10-hexadecenoic acid content increased as the temperature was reduced from 35 degrees C to 26-20 degrees C . At lower temperatures tuberculostearic acid decreased while oleic and linoleic acids increased, the latter being found in M . phlei for the first time . Concomitantly palmitic acid content decreased, and the 6- and 9-hexadecenoic acids increased slightly on reducing the temperature from 35 to 10 degrees C . Thus, down to 26-20 degrees C palmitic acid was mainly replaced by 10-hexadecenoic acid . From this range down to 10 degrees C, palmitic and tuberculostearic acids were replaced by oleic and linoleic acids . Consequently, fatty acid branching decreased and mean chain length increased, as the temperature was reduced . These observations support the view that regulation of membrane fatty acid composition is part of microbial temperature adaptation, and that the mechanism behind the responses might be more complex than generally believed.

Br J Ophthalmol, 1993 Jan, 77(1), 41 - 5
Microbial contamination of contact lens cases in the west of Scotland; Devonshire P et al.; A cross-sectional study of 178 asymptomatic contact lens wearers attending 10 contact lens practices in the west of Scotland was conducted over a 4 month period . The aims of the study were to identify specific microbial contaminants in lens cases, to determine the rate of contamination of such containers and to assess the value of the steps involved in different lens care regimens in the prevention of case contamination . Microbial contamination affected 53% of lens cases . Cases used with conventional wear and disposable systems were contaminated at similar rates and, therefore, the advantage of regular lens replacement may have been lost if these lenses were stored in contaminated cases . Four percent of lens cases were contaminated with amoebal species and all of these showed concomitant bacterial colonisation . These findings imply that case hygiene is probably as important as lens hygiene if new or disinfected lenses are not to be immediately re-contaminated by storage in dirty cases . Unfortunately simple and effective methods of lens and case disinfection, which would be suitable for use in the average home environment, are not yet available . It follows that frequent and regular disposal of lens cases may prove to be a necessary measure to prevent the build-up of microbial colonisation in such containers.

Adv Enzymol Relat Areas Mol Biol, 1993, 66, 149 - 201
Thioltransferases; Wells WW et al.; A family of small molecular weight proteins with thiol-disulfide exchange activity have been discovered, widely distributed from E . coli to mammalian systems, called thioltransferases or glutaredoxins . There are no substantiated reports of thioltransferases-glutaredoxins in plants; however, partially purified dehydroascorbate reductase from peas had thiol-disulfide exchange catalytic activity using glutathione as reductant and S-sulfocysteine as thiosulfate cosubstrate (unpublished data) . Thus, this class of proteins is universally distributed . Based on mutagenesis studies, a sequence of Cys-Pro-Tyr(Phe)-Cys- followed by Arg-Lys- or Lys alone is critical for both the thiol-disulfide exchange reaction and the dehydroascorbate reductase activity . The dithiol-disulfide loop represented by this structure is unique since the cystine closer to the N-terminus has a highly acidic thiol pKa (3.8 as determined for the pig liver enzyme) that contributes to the protein's high S- nucleophilicity . Compared with the microbial enzyme, the mammalian thioltransferases (glutaredoxins) are extended at both N and C termini by 10-12 amino acid residues, including a second pair of cysteines toward the C-terminus with no known special function . Yeast thioltransferase is more like mammalian enzymes in length (106 amino acids) but more like E . coli glutaredoxin in being unblocked at the N-terminus and having only one set of cysteines; that is, at the active center . The three mammalian enzymes, for which sequences are available, are blocked at the N-terminus by an acetyl group linked to alanine with no known special function other than possibly to impart greater cellular turnover stability . A report of carbohydrate (8.6%) content in rat liver thioltransferase has not been verified by more sensitive methods of carbohydrate analysis, nor has carbohydrate been identified in samples of purified glutaredoxin from any source . Thiol transferase and glutaredoxin are two names for the same protein based on similarity of amino acid sequence, immunochemical cross-reactivity, and other enzyme properties . The inability of thioltransferase from some mammalian sources to act as an electron carrier in ribonucleotide reductase systems, whether homologous or heterologous in origin, remains to be explained in future studies.

Poult Sci, 1993 Jan, 72(1), 137 - 43
An in vitro assay for prediction of broiler intestinal viscosity and growth when fed rye-based diets in the presence of exogenous enzymes; Bedford MR et al.; A two-stage in vitro assay procedure was developed for predicting the efficacy of microbial enzyme sources in rye-based diets for broiler chicks . The procedure uses the complete diet as substrate to predict the in vivo intestinal viscosity and final weight of birds fed such diets . The optimum conditions for maximizing in vitro viscosity of a rye-based diet consisted of digesting .6 g of diet (ground through a 1-mm screen) in .9 mL of .1 N HCl containing 2,000 U pepsin/mL for 45 min at 40 C . At the end of this incubation phase, .3 mL 1 M NaHCO3 containing 8 mg pancreatin/mL (8 x USP) was added and the tube sealed and incubated for 2 h with intermittent vortexing . The digesta was then centrifuged (12,700 x g) and the supernatant collected for viscosity analysis using a digital viscometer . This assay was used for a rye-based diet containing either 0, .1, .2, .4, .8, or 1.6% of a xylanase source (experimental Trichoderma longibrachiatum product) . The results were compared to weight and intestinal viscosity proximal and distal to Meckel's diverticulum of broilers (19 days of age) that had been fed these diets . The in vitro assay accurately predicted the in vivo intestinal viscosity (proximal r2 = .758, P < .0001, distal r2 = .667, P < .0001) and final weight of these birds (r2 = .660, P < .0001) . The data suggest that the in vitro assay is a reliable assay for assessing the growth-promoting ability of an enzyme preparation in chicks fed rye-based diets.

Clin Chem, 1993 Jan, 39(1), 125 - 8
Discordant results for determinations of triglycerides in pig sera; Tuten T et al.; We recently determined triglyceride concentrations in pig sera by three fully enzymatic methods (Kodak Ektachem 700, Hitachi 707, and Abbott EPx) and obtained significantly lower values than those obtained with chemical or enzymatic methods based on chemical hydrolysis . All methods used involve microbial lipases for liberating glycerol from glycerides and glycerol phosphate dehydrogenases or oxidases for subsequent oxidation . The methods were validated against reference methods by using fresh human sera and survey materials . The discordant results were not from matrix sample-method interaction but from incomplete hydrolysis of pig serum triglycerides by the lipolytic enzymes . When serum triglycerides from 10 pigs showing the highest biases were hydrolyzed by microbial lipases and the reaction mixture was subjected to thin-layer and gas-liquid chromatography, the predominant end products were palmitoyl monoglyceride and a mixture of free fatty acids with the following composition (fatty acid as percent of total +/- SD): 16:0, 7.8 +/- 2; 18:0, 5.4 +/- 2.2; 18:1, 53 +/- 12; 18:2, 31 +/- 4.6; and 18:3, 2.5 +/- 1 . Assuming that the lipases exhibit the usual specificity toward the 1 and 3 positions of the triglyceride, the data suggest that, in pig, triglycerides 18:1 and 18:2 occupy the 1 and 3 positions and 16:0 (palmitic acid) predominantly occupies the 2 position . Triglycerides of this structure may not be well hydrolyzed by the typical lipolytic enzymes in clinical assays.

Infect Immun, 1993 Jan, 61(1), 303 - 6
Apathogenic, intestinal, segmented, filamentous bacteria stimulate the mucosal immune system of mice; Klaasen HL et al.; Segmented filamentous bacteria (SFBs) are apathogenic autochthonous bacteria in the murine small intestine that preferentially attach to Peyer's patch epithelium . SFBs have never been cultured in vitro . We have studied the effects of SFBs on the immune system of the host . Mice monoassociated with SFBs were compared with germ-free mice and with mice without SFBs but with a specific-pathogen-free (SPF) gut flora . SFBs versus no microbial flora raised the number of lymphoid cells in the lamina propria of the ileal and cecal mucosa, raised the number of immunoglobulin A (IgA)-secreting cells in the intestinal mucosa, produced elevated IgA titers in serum and intestinal secretions, and enhanced the concanavalin A-induced proliferative responses of mesenteric lymph node cells . The SPF flora had effects similar to but less pronounced than those mediated by SFBs . The results indicate that SFBs stimulate the mucosal immune system to a greater extent than do other autochthonous gut bacteria.

Am J Respir Cell Mol Biol, 1993 Jan, 8(1), 20 - 7
Interleukin-8 expression in normal nasal epithelium and its modulation by infection with respiratory syncytial virus and cytokines tumor necrosis factor, interleukin-1, and interleukin-6; Becker S et al.; Inflammation in nasal and airway tissue caused by allergens, microbial infection, and air pollution are likely to be regulated by inflammatory mediators produced by airway epithelial cells . We have therefore investigated the baseline expression of a number of cytokine genes known to be important inducers and modulators of inflammation, in freshly isolated human nasal epithelium . Cells were obtained by superficial scraping of turbinate tissue, and cDNA for polymerase chain reaction (PCR) amplification was reverse-transcribed directly from lysates of 3 x 10(3) to 5 x 10(3) epithelial cells using random hexamers . Constitutive expression of relatively high levels of interleukin-8 (IL-8) mRNA but undetectable levels (< 1 mRNA copy/cell) of granulocyte/macrophage colony-stimulating factor (GM-CSF), IL-6, IL-1, or tumor necrosis factor (TNF) mRNA were found after PCR amplification of the cDNA . IL-8 protein, but not IL-6, was identified in the nasal epithelial cells by immunocytochemistry . Infection with respiratory syncytial virus (RSV) or stimulation of nasal epithelium for 4 h with TNF or IL-1 in vitro resulted in a 4- to 10-fold increase in IL-8 mRNA expression but not in the expression of detectable levels of mRNA for the other cytokines . IL-8 was secreted by RSV-, IL-1-, and TNF-stimulated as well as unstimulated nasal epithelial cells after 6 to 20 h of culture . Neither IL-6, GM-CSF, nor TNF activity/immunoreactivity was detectable in the culture supernatants . Thus, it appears that IL-8 is a major cytokine of human nasal epithelium, constitutively expressed and readily secreted upon virus infection or stimulation with IL-1 and TNF.

Life Sci, 1993, 52(1), 73 - 84
Immunostimulatory factor(s) in the porcine uterine fluids; Li WI et al.; The effects of porcine uterine fluid filtrate (PUF) on lymphocyte activity were examined . PUF was added to porcine lymphocyte cultures in the presence or absence of phytohemagglutinin (PHA), concanavalin A (Con A), or pokeweed mitogen (PWM) . Lymphocyte response was determined by 3H-thymidine incorporation . Results indicated that PUF enhanced PHA-, Con A-, and PWM-induced lymphocyte mitogenesis by 222%, 207%, and 261%, respectively, as compared to control . The stimulatory effect of PUF was unlikely to be due to the presence of interleukin-2 in PUF, by evidence that PUF alone did not support cytotoxic T-lymphocyte proliferation, nor was the stimulatory effect due to microbial contamination, because treatment of PUF through PyroBind columns to remove endotoxin did not affect its activity . Incubation of PUF at 56C for 40 min did not alter the enhancement effect . Treatment of PUF with trypsin or pronase for up to 24 hr did not change its activity . In mixed lymphocytes reaction, PUF augmented 3H-thymidine uptake by 242% . The molecular (mol) mass of the immunostimulatory activity in PUF was examined by size-exclusion HPLC, Sephadex G-50, and G-15 gel filtration, and it was found to be 0.8 kDa and < 0.5 kDa . Extracts of porcine endometrium also enhanced PHA-induced lymphocyte mitogenesis, suggesting that the factor(s) may be of uterine origin . In conclusion, we have identified an factor(s) in PUF capable of stimulating lymphocyte mitogenesis and mixed lymphocyte reaction.

Folia Histochem Cytobiol, 1993, 31(2), 77 - 81
Cell proliferation in colonic crypts of germ-free and conventional mice--preliminary report; Nowacki MR; The influence of normal microbial flora on cell proliferation in mouse colonic crypts was assessed by a comparison of conventional and germ-free animals . The following characteristics of the crypts were measured: length, diameter in the middle, number of columns, number of cell per one column, percentage of goblet cells, mitotic and labelling index distribution and average cell cycle time for the proliferative compartment . It has been found that the crypts in conventional mice are longer than in their germ-free counterparts, while their diameter and percentage of goblet cells were similar in both animal groups . The mitotic index was lower in germ-free mice, as well as the labelling index . The average cell cycle time was about 1.7-fold shorter in conventional animals than in germ-free animals . This result was verified using computer simulation model; it turned out that the change in average cell cycle time could be solely responsible for all the observed differences between germ-free and conventional animals . It seems therefore that normal microbial flora stimulates cell proliferation in mouse colon, most probably due to shortening of the average cell cycle.

J Anim Sci, 1993 Jan, 71(1), 38 - 43
Effects of hybrid and ensiling with and without a microbial inoculant on the nutritional characteristics of whole-plant corn; Hunt CW et al.; Two corn hybrids (Pioneer 3377 and 3389) with similar total plant and grain yield characteristics were evaluated to determine potential differences in nutritive value of the whole plant and stover . Hybrids were grown in plots at four different locations in Idaho in 1988 and 1989 for laboratory evaluation . Samples from the plots were stored fresh or ensiled with or without a microbial inoculant for 60 d in laboratory silos . Whole-plant samples of 3377 had a lower (P < .01) percentage of NDF (42.7 vs 48.1), ADF (26.3 vs 30), hemicellulose (16.2 vs 18.2), cellulose (21.4 vs 24), and lignin (3.4 vs 3.8) . Similar differences, but of larger magnitude, were observed in stover samples . Ruminal in situ (24 h) and two-stage IVDMD of whole-plant and stover samples were greater (P < .01) for 3377 than for 3389 . Lower fiber content and greater degradability for 3377 was not due to greater grain content; 3377 actually had numerically lower grain as a percentage of whole plant than 3389 (41 vs 44) . Ensiling reduced (P < .05) NDF, hemicellulose, and cellulose content compared with fresh samples; however, no effect (P > .10) due to inoculant was noted . Hybrid x storage interactions were not observed (P > .10) . Inoculation and hybrid did not affect (P > .10) organic acid concentration of the silages . In 1989 silages from 3377 and 3389 were harvested to evaluate diets containing 65% (DM basis) corn silage in steer digestion and growth trials.(ABSTRACT TRUNCATED AT 250 WORDS)

J Anim Sci, 1993 Jan, 71(1), 213 - 7
Influence of oral antibiotics on digestive function in Holstein steers fed a 71% concentrate diet; Zinn RA; Four Holstein steers (164 kg) with cannulas in the rumen and proximal duodenum were used to evaluate the effects of oral antibiotic administration on characteristics of digestion of a 71% concentrate diet . Treatments were as follows: 1) no antibiotic; 2) 350 mg.steer-1.d-1 of chlortetracycline (CTC) provided in the feed (A350); 3) 350 mg.steer-1.d-1 of CTC plus 350 mg.steer-1.d-1 of sulfamethazine provided in the feed (AS700); 4) 22 mg of CTC/kg BW (3.6 g.steer-1.d-1) provided in the drinking water (A3600) . Drinking water intake averaged 23.5 L/d and was not affected (P > .10) by treatments . Antibiotic treatments increased passage (P < .05) of OM to the small intestine . This effect was largely due to decreased (27%, P < .05) ruminal ADF digestion . Passage of nonammonia N to the small intestine was greater (10.4%, P < .01) for A3600 vs A350 and AS700; the latter was similar to the control . This effect was due to an increased (23.4%, P < .05) synthesis of microbial N . Total tract digestion of OM (P < .05), starch (P < .05), N (P < .10), and DE (P < .10) were slightly lower (2.7, 1.6, 2.8, and 2.7%, respectively) for antibiotic-supplemented diets . Total tract starch (P < .10) and N (P < .01) digestion were further decreased (1.2 and 3.5%, respectively) for A3600 vs A350 and AS700 . Postruminal digestion of ADF was greater (131%, P < .10) for antibiotic-supplemented diets, compensating for the lower ruminal digestion . There were no treatment effects (P > .10) on ruminal pH.(ABSTRACT TRUNCATED AT 250 WORDS)

J Am Dent Assoc, 1993 Jan, 124(1), 48 - 56
Cleaning, sterilization and disinfection: basics of microbial killing for infection control; Miller CH; Cleaning items directly involved in patient care makes the disinfection or sterilization process easier . Specific steps in cleaning instruments and equipment and dental unit surfaces are described . Recommended infection control procedures are listed, together with monitoring methods.

Mol Aspects Med, 1993, 14(1), 1 - 81
Human immune toxicity; Burrell R; This review examines xenobiotic toxicity to the immune system, stressing in particular those aspects of most relevance to humans . Immunotoxicity is examined especially from three points of view: by what immunological component is affected, by classes of foreign agents that adversely affect the human immune system and by critical evaluation of human case reports and epidemics . Mechanisms by which xenobiotics interrupt cytokine networks are emphasized . The concept that microbial agents, both environmental as well as infectious, may act as immunotoxicants, either alone or in synergism with conventional agents is introduced . Instances of human immunotoxicology are critically evaluated in terms of clinical relevance, i.e . whether increased susceptibility to opportunistic infections or tumor emergence takes place in the affected individuals.

Dig Dis, 1993, 11(3), 146 - 56
The gastrointestinal mucosal immune system; James SP; The mucosal immune system, consisting of lymphoid tissues associated with the lacrimal, salivary, gastrointestinal, respiratory and urogenital tracts and lactating breasts, quantitatively contains the majority of the lymphoid tissue of the body . There are a number of important features of the gastrointestinal mucosal immune system: the mucosal immune system contains specialized structures, such as the Peyer's patches, where immune responses are thought to be initiated; there is a pattern of relatively specific recirculation of lymphoid cells to the mucosa, known as mucosal homing; subsets of lymphoid cells, particularly IgA B cells and memory T cells, predominate at mucosal surfaces; and the predominant mucosal immunoglobulin, secretory IgA, is particularly well adapted to host defense at mucosal surfaces . These elements of the gastrointestinal mucosal immune system function together to generate an immune response which on the one hand protects the host from harmful pathogens, but on the other hand is tolerant of the ubiquitous dietary antigens and normal microbial flora.

Gerontology, 1993, 39 Suppl 1, 30 - 7
Purification of a trypsin-type protease from human umbilical vein endothelial cells which is highly sensitive to the Kunitz inhibitor domain peptide of Alzheimer's disease amyloid protein precursor; Kido H et al.; A trypsin-type protease was purified to enzymatic homogeneity from human umbilical vein endothelial cells by sequential affinity chromatographies . The enzyme specifically hydrolyzed dibasic substrates with leucine at the P3 positions, but scarcely hydrolyzed the other substrates tested . The enzyme was strongly inhibited by the Kunitz inhibitor domain peptide of Alzheimer's disease amyloid protein precursor (Ki value, 0.35 nM) and by the microbial inhibitors leupeptin and anti-pain . These results, together with a previous finding of a significant increase in the expression of Alzheimer's amyloid protein precursors (beta APPs) with the Kunitz inhibitor domain in Alzheimer's disease, suggest that the activity of the trypsin-type protease is suppressed by an increase of beta APPs with inhibitor activity in Alzheimer's disease, resulting in aberrant intracellular protein catabolism including degradation of beta APPs and beta-protein deposition.

Appl Biochem Biotechnol, 1993 Jan-Feb, 38(1-2), 141 - 6
High-throughput microtiter plate-based chromogenic assays for glycosidase inhibitors; Walker JM et al.; Rapid microtiter plate-based colorimetric assays have been developed that allow the screening of large numbers of samples for the presence of inhibitors of alpha-glucosidase, alpha-amylase, and beta-galactosidase . The assays are particularly useful for screening large numbers of microbial culture filtrates.

Ann Biol Clin (Paris), 1993, 51(1), 1 - 6
{Contribution of flow cytometry in virology}; Wattre P; The development of commercial multichannel air-cooled laser flow cytometers which allow measurement of one or more characteristic parameters of particles or cells has been furthered due to advances in probes and fluorochromes . FITC-conjugated antibodies specific for selected cell surface molecules or for viral antigen expression in infected cells, fluorescein diacetate for labeling cells as a parameter of viability and propidium iodide as a DNA stain are generally used . Flow cytometric assays have been developed in virology to study the interactions of viruses with the immune system (Hantan Virus), to evaluate the diagnosis of secondary immunodeficiency syndromes (HIV), to analyze the protein and DNA content of virally infected cells during the cell cycle (SV40), to detect immediate early, early or late antigens in HCMV-infected cells, to assess the HCMV persistence in mononuclear cells, for use as a model for studying virus attachment to cellular receptors (EBV-echovirus), to screen and evaluate antiviral agents (Influenza C, HCMV, HSV, HIV) and to quantify antigens or antibodies simultaneously against various viruses (HCMV and HSV) or against different antigenic glycoproteins (HIV) . This technology has thus become an important tool for the clinical virology and microbial serology laboratories.

Br Med Bull, 1993 Jan, 49(1), 1 - 16
The British Medical Bulletin 1943-1993 . A guide to medical science and thought in Britain; Tansey EM et al.; Established 50 years ago by The British Council, to promote medical science abroad, the British Medical Bulletin (BMB) has evolved from a stencilled list of abstracts into a distinguished medical periodical . It has reflected the extraordinary diversity of advances that have been made in medicine in this country . During these 5 decades anti-microbial chemotherapy has become firmly established; organ and tissue transplantation have become a reality; the significance of Watson and Crick's discovery of the structure of DNA has been realised by the increasing applications of molecular biology to human disease; and modern investigative techniques, particularly magnetic resonance imaging, have made the human body virtually transparent . At the same time the BMB has mirrored fresh concerns, with, for example, industrial and environmental hazards, with newly recognised infectious agents, and increasingly with the medico-social problems of the modern era . This paper reviews the scientific contributions of the Bulletin and some of the administrative and financial structures that have supported it.

Appl Biochem Biotechnol, 1993 Spring, 39-40, 739 - 52
Microbial reduction of sulfur dioxide with pretreated sewage sludge and elemental hydrogen as electron donors; Deshmane V et al.; It has been demonstrated tht heat- and alkali-pretreated-sewage sludge may serve as an electron donor and carbon source for SO2 reduction by Desulfovibrio desulfuricans . A continuous D . desulfuricans culture was operated for 6 mo with complete reduction of SO2 to H2S . The culture required only minor amounts of mineral nutrients in addition to pretreated sewage sludge . It has also been shown that the sulfate-reducing bacterium Desulfotomaculum orientis can be grown on H2 as an energy source, CO2 as a carbon source, and SO2 as a terminal electron acceptor . Complete reduction of SO2 to H2S was observed.

Ann Chir Gynaecol, 1993, 82(1), 51 - 4
Bacterial contamination and closed suction drainage in open meniscectomy of the knee; Lindahl J et al.; A prospective study was undertaken to investigate the occurrence and possible sources of microbial contamination in association with open meniscectomy of the knee . Sixty patients were randomized into three closed suction drainage regimens of 12, 24 and 48 hours' postoperative suction drainage, respectively . Specimens for bacterial culture were taken from the joint at the time of surgery, from the surgical suction tips, from the subcutaneous part of the drain surface and from the drain-contained fluid at the time of drain removal . The airborne bacterial contamination showed no marked role in this bacterial invasion into the joint, nor did the use of suction increase the risk . The vacuum suction drain, on the other hand, appeared to carry an obvious risk of bacterial invasion, as a parallel of time . In the present comparison this risk was minimal when the drain was removed 12 hours postoperatively.

Antonie Van Leeuwenhoek, 1993, 63(3-4), 299 - 313
Growth kinetics and competition--some contemporary comments; Gottschal JC; Results of competition experiments with one growth-limiting factor under idealized experimental conditions have been reported extensively, and usually provide ample support for the conclusion that 'complete competitors cannot coexist' . However, under conditions of multiple substrate limitation and discontinuous or alternating supply of nutrients, coexistence of species is quite common . Since such patterns of nutrient supply may be expected to prevail in many natural environments the mechanisms ruling the survival and growth of bacteria under such conditions need to be understood . However, it appears that surprisingly little is known of the physiological state of individual competing species grown in mixed cultures . Unfortunately, basic information such as the actual concentration of limiting nutrients is lacking in most cases . But perhaps the recent development of new and powerful techniques to explore the physiological properties even of individual cells will further stimulate studies into the mechanisms behind the competitiveness of microbial species.

Antonie Van Leeuwenhoek, 1993, 63(3-4), 243 - 74
Concentrations and fluxes of organic carbon substrates in the aquatic environment; Munster U; Data concerning concentrations and fluxes of dissolved organic compounds (DOC) from marine and lacustrine environments are reviewed and discussed . Dissolved free amino acids and carbohydrates comprised the main fraction in the labile organic carbon pool . Dissolved free amino acids in marine waters varied between 3-1400 nM and those of fresh waters between 2.6-4124 nM . Dissolved free carbohydrates varied between 0.4-5000 nM in marine systems and between 14-111 nM in fresh waters, The turnover times of both substrate pools varied in marine waters between 1.4 hours and 948 days and in fresh waters between 2 hours and 51 days . Measurements of stable 12/13C-ratio and 14C-isotope dating in ocean deep water samples revealed DOC turnover times between 2000-6000 years . Studies on carbon flows within the aquatic food webs revealed that about 50% of photosynthetically fixed carbon was channelled via DOC to the bacterioplankton . Excreted organic carbon varied between 1-70% of photosynthetically fixed carbon in marine waters and between 1-99% in fresh waters . The labile organic carbon pool represented only 10-30% of the DOC . The majority (70-90%) of the DOC was recalcitrant to microbial assimilation . Only 10-20% of the DOC could be easily chemically identified . Most of the large bulk material represented dissolved humic matter and neither the chemical structure nor the ecological function of the DOC is as yet clearly understood.

Antonie Van Leeuwenhoek, 1993, 63(3-4), 225 - 35
Nutrient-limited microbial growth kinetics: overview and recent advances; Button DK; Traditional concepts of nutrient uptake and growth kinetics as linked by cell yield are presented . Phenomena affecting the kinetics are examined along with a discussion of those which lead to ambiguity . Concepts of flux control are presented to help understand the distribution of material along metabolic pathways . Specific affinity is described to relate nutrient accumulation rates to transporter density . It is shown to be a primary kinetic constant and the best available index of nutrient collection ability . As an aid to understanding, specific affinity is reexpressed in terms of membrane permeability . Formulations of nutrient transport rate as a function of cellular composition, particularly transporter and enzyme content and known as janusian kinetics, are described as an improvement to specific affinity theory . Procedures for quantified unidirectional fluxes are reviewed to identify the difference between gross and net transport rates of substrate . Collision frequency theory is used to show that in addition to total biomass, cell size and transporter density should also be included in rate equations describing microbial growth . Theory diversity suggests that one reason for microbial metabolic is that the likelihood of additional collisions of substrate molecules with a cell surface, after an initial collision, requires only a sparse distribution of transporter sites for maximal rate, leaving room for additional transporters able to collect other substrate types.

Rev Inst Med Trop Sao Paulo, 1993 Jan-Feb, 35(1), 1 - 10
{Legionellosis associated with pneumopathies in São Paulo . Study of the etiologic confirmation by isolation and serology}; Mazieri NA et al.; The role of Legionella sp as an important pathogen, although reported in many countries, had not been well documented in Brazil . The main objective of the present study is to detect this organism or it's immunological response in patients with pulmonary infections in the city of Sao Paulo . For this purpose, specific laboratory methodology was introduced to cultivate the agent and demonstrate specific antibodies by serology . Patients from two University centers in Sao Paulo were studied: 100 from the Hospital Universitario, University of Sao Paulo with general pulmonary infections and 100 from Hospital das Clinicas, Faculdade de Medicina, University of Sao Paulo . The latter were selected to exclude pulmonary infections of other bacterial aetiology, and 30 of these were selected from the Renal Transplant Unit . Clinical specimens for cultures included sputum tracheal secretion, pleural, fluid, bronchial washing or lung tissue biopsy . Isolation of Legionella sp was attempted in BCYE medium with and without antibiotics, presumptive diagnosis by biochemical methods and identification through direct immunofluorescence staining and slide agglutination test . Direct evidence of the organism in tissue was attempted by direct immunofluorescence staining . Specific antibodies for Legionella pneumophila serogroup 1 were tested by the indirect immunofluorescence assay . Serology was also carried out in specimens from human contacts with Legionnaires' Disease . Legionella pneumophila serogroup 1 was isolated from two patients, demonstrating the presence of the pathogen in this country . Serology was able to establish present or past infection with the agent in 16 of the 100 patients from Hospital das Clinicas and in only one from Hospital Universitario . In patients considered as high risk groups from Hospital das Clinicas with renal transplantation, serology for present or past Legionellosis was positive in 33% and in 8.5% for community acquired infections . Serology in specimens from Medical personnel in contact with patients of Legionnaires' disease was positive for past infection in one single subject, confirming information already published that direct transmission from person to person is unusual . The introduction of specific methods for laboratorial evidence of Legionella sp infections at the Hospital das Clinicas-Faculdade de Medicina USP community has permitted diagnosis in able time to allow use of specific anti-microbial therapy, with good results.

J Cancer Res Clin Oncol, 1993, 120(1-2), 5 - 11
Stimulation of prostaglandin E2 production and induction of specific protein synthesis in rat peritoneal macrophages by a tumor promoter staurosporine; Tamura T et al.; Staurosporine is a microbial anti-fungal alkaloid having potent inhibitory activity on protein kinase C and is a non 12-O-tetradecanoylphorbol-13-acetate-type tumor promoter in two-stage carcinogenesis experiments in mouse skin . Effects of staurosporine and its structurally related compounds K-252a, KT5720 and KT5822 on prostaglandin E2 production, release of arachidonic acid from membrane phospholipids, and uptake of {35S}methionine into intracellular proteins were examined in rat peritoneal macrophages . Among the four compounds, only staurosporine stimulated the production of prostaglandin E2 and release of arachidonic acid at concentrations of 1 ng/ml and 10 ng/ml . The uptake of {35S}methionine into cellular proteins, estimated to be 120 kDa and 125 kDa molecular mass, was also stimulated by staurosporine treatment, and the uptake was increased in parallel with the increase in prostaglandin E2 production . At higher concentrations (100 ng/ml and 1000 ng/ml), staurosporine inhibited prostaglandin E2 production and did not induce the specific protein synthesis . Other compounds neither stimulated prostaglandin E2 production nor induced specific protein synthesis . K-252a inhibited prostaglandin E2 production at concentrations above 10 ng/ml . These results suggest that the staurosporine-induced proteins might participate in the tumor promotion or at least in the staurosporine-induced stimulation of prostaglandin E2 production.

Int J Periodontics Restorative Dent, 1993, 13(3), 213 - 43
Routine anaerobic bacterial culture and systemic antibiotic usage in the treatment of adult periodontitis: a 6-year longitudinal study; Rosenberg ES et al.; This investigation evaluated the efficacy of obtaining baseline culture and sensitivity data on a routine basis from the patient with adult periodontitis . Patients diagnosed with chronic adult periodontitis, rapidly progressive periodontitis, or refractory periodontitis were followed for up to 6 years . More than 95% of patients with chronic adult periodontitis were successfully treated with mechanical therapy alone . Approximately one half of the patients with rapidly progressive periodontitis were treated successfully without antibiotics . All of the patients with refractory periodontitis required systemic antibiotics as part of treatment . Most patients with chronic adult periodontitis exhibited one or two species of organisms at baseline, and these organisms were eliminated or reduced to low levels by mechanical therapy . In contrast, patients with rapidly progressive or refractory periodontitis consistently demonstrated multiple species and required systemic antibiotics in conjunction with mechanical therapy to alter the subgingival microbial milieu . Routine culturing and antibiotic therapy is contraindicated in patients with chronic adult periodontitis, but may be beneficial for successful treatment of patients with rapidly progressive or refractory periodontitis.

Annu Rev Microbiol, 1993, 47, 57 - 87
Agroactive compounds of microbial origin; Tanaka Y et al.; Microbial metabolites attract increasing attention as potential pesticides . They are expected to overcome the resistance and pollution that have accompanied the use of synthetic pesticides . Several microbial metabolites, such as avermectin, have proved useful as agroactive agents . In this review, we attempt to identify newer agroactive microbial metabolites with feasible activity or interesting action sites from those reported in recent years . In addition, microbial and chemical modifications of existing microbial agrochemicals are discussed to illustrate the usefulness of these technologies in potentiating agroactivity and stability . We discuss the possibility of future discovery of excellent microbial agrochemicals, and the importance of efforts to promote positive public perception and public acceptance of pesticide chemicals.

Haematologia (Budap), 1993, 25(2), 69 - 84
Adequate iron stores and the 'Nil nocere' principle; Hollan S et al.; There is a need to change the policy of unselective iron supplementation during periods of life with physiologically increased cell proliferation . Levels of iron stores to be regarded as adequate during infancy and pregnancy are still not well established . Recent data support the view that it is not justified to interfere with physiological adaptations developed through millions of years by sophisticated and precisely coordinated regulation of iron absorption, utilization and storage . Recent data suggest that the chelatable intracellular iron pool regulates the expression of proteins with central importance in cellular iron metabolism (TfR, ferritin, and erythroid 5-aminolevulinic synthetase) in a coordinately controlled way through an iron dependent cytosolic mRNA binding protein, the iron regulating factor (IRF) . This factor is simultaneously a sensor and a regulator of iron levels . The reduction of ferritin levels during highly increased cell proliferation is a mirror of the increased density of TfRs . An abundance of data support the vigorous competition for growth-essential iron between microbial pathogens and their vertebrate hosts . The highly coordinated regulation of iron metabolism is probably crucial in achieving a balance between the blockade of readily accessible iron to invading organisms and yet providing sufficient iron for the immune system of the host . The most evident adverse clinical effects of excess iron have been observed in immunodeficient patients in tropical countries and in AIDS patients . Excess iron also increases the risk of initiation and promotion of malignant processes by iron binding to DNA and by the iron-catalysed release of free radicals . Oxygen radicals were shown to damage critical biomolecules leading, apart from cancer, to a variety of human disease states, including inflammation and atherosclerosis . They are also involved in processes of aging and thrombosis . Recent clinical trials have suggested that the use of iron-chelators, natural and synthetic antioxidants, and anti-TfR monoclonal antibodies can contribute in retarding malignant cell proliferation . Hypoferraemia during pregnancy is--like haemodilution--an adaptation to the risks involved in the natural hypercoagulable state of pregnancy . It may also serve to prevent the risk of infections and mutagenicity in the highly proliferating tissues of the foetus . Blunted erythropoiesis has been revealed during the first 30 weeks of pregnancy by the use of the newly developed method of determining the soluble serum transferrin receptor . The lack of increase in erythropoietin levels proves that there is no hypoxia . Decreases in Hb and iron levels are parts of a physiological adaptation . As a consequence they should neither be treated nor prevented . It is stressed that whenever a widespread and ingrained routine medical intervention has to be changed it is essential to first monitor the potential health effects of the recommended change in a national policy.

Rev Environ Contam Toxicol, 1993, 133, 1 - 58
Environmental fate and effects of dicamba: a Canadian perspective; Caux PY et al.; Literature on the environmental fate and effects of the benzoic acid herbicide dicamba was reviewed to provide a scientific basis to derive Canadian Water Quality Guidelines . Included in the review was information on the uses and production of dicamba, its physical and chemical properties, environmental monitoring data in Canadian surface water and groundwater, soils, sediments, and biota, and its environmental degradation, persistence, and fate . Through monitoring, dicamba has been detected in less than 8% of surface-water samples to a maximum concentration of 13 micrograms.L-1, while 2% of groundwater samples were positive up to 517 micrograms.L-1 . Only one study that analyzed sediments (with no detections) and no field studies that investigated residues in biota were found . Microbial degradation is the most important process governing the dissipation of dicamba in aquatic and soil environments . Photolysis, hydrolysis, volatilization, adsorption to sediment, and bioconcentration are not expected to be significant removal processes, based on limited environmental fate data . The half-life of dicamba in water is < 7 d, although residues have been detected in surface-water supplies in Alberta more than 6 mon after application . The literature reports the half-life in soils ranges from 4 to 555 d; however, < 12 wk would be typical under Canadian conditions . High moisture and temperature, and other conditions that favor microbial degradation, would likely reduce the half-life to < 4 wk . The principal soil and plant metabolite is 3,6-dichlorosalicylic acid, with minor amounts of 2,5-dihydroxy-3,6-dichlorobenzoic acid and 5-hydroxydicamba found . Dicamba is highly mobile in soil, and significant leaching is possible; its water solubility is 6.5 g.L-1 (25 degrees C) and it has a log octanol-water partition coefficient of 0.477 . Acute and chronic toxicological studies for all nontarget plants and animals were also reviewed . The major groups of organisms for which toxicological data were collected were freshwater fish, invertebrates and plants, tame hays and cereals, legumes, and other crops, and livestock poultry and mammals . The acute toxicity (< or = 96-hr LC50) to freshwater fish ranged from 28 to 516 mg.L-1, whereas that for invertebrates ranged from 3.9 to > 100 mg.L-1 . No chronic data were found for either of these groups . The chronic EC50 to 14 freshwater algae, based on growth inhibition, ranged from 100 to > 10,000 micrograms.L-1 . No studies on freshwater macrophytes or any marine organisms were found . Agricultural crops exhibited varying toxicity.(ABSTRACT TRUNCATED AT 400 WORDS)

J Orthop Trauma, 1993, 7(5), 417 - 27
Fracture blisters: clinical and pathological aspects; Varela CD et al.; Fracture blisters are tense vesicles or bullae that arise on markedly swollen skin directly overlying a fracture . There is very little objective data in the literature detailing their characteristics and management . All fracture blisters that occurred over a 3 1/2-year period were studied retrospectively at four hospitals, of which three were level I trauma centers . A total of 53 blisters developed in 51 patients . They occurred in characteristic locations along the human musculoskeleton, most commonly overlying the tibia, ankle, and elbow . They arose within 24-48 h of acute injury in most instances . The timing of surgical intervention affected the occurrence of fracture blisters . Those patients with acute fractures who underwent open reduction internal fixation (ORIF) within 24 h of injury had the lowest incidence of fracture blisters (2.0%) compared with those delayed for > 24 h (8.0%) (p < 0.001) . In those patients with fracture blisters present at time of surgery, patient care was affected in 10 of 13 cases (71%) . Two of these were major complications occurring as postoperative wound infections . Other management problems consisted of delaying surgery, and changing in the operative plan . There were no adverse affects on patient care when the fracture blister developed postoperatively . Twenty-one fractures with blisters were treated by closed means, with the presence of fractures blisters delaying closed reduction and casting in two . Biopsy examination of 15 blisters supported the clinical impression that fracture blisters are subepidermal vesicles . The blister fluid was found to be a sterile transudate . Microbial evaluation of 11 ruptured fracture blisters demonstrated colonization (primarily with skin pathogens), occurred soon after blister rupture, and continued until reepithelialization.

Biol Cell, 1993, 78(1-2), 123 - 8
Application of flow cytometry to rapid microbial analysis in food and drinks industries; Laplace-Builhe C et al.; In food and drinks industries, the time required for conventional tests can lead to substantial delays in product release to the market . Flow cytometry (FCM) has been used in conjunction with viability markers for rapid counting of yeast, mould and bacterial cells in food products . A single-parameter flow cytometer has proved applicable to the rapid detection of low numbers of microbial contaminants in finished products . The excellent correlation between FCM results and product quality shelf-life expiry date has allowed the establishment of realistic quality control criteria for rapid positive release of product . Used for the monitoring of microbial biomass during manufacturing processes, flow cytometry allowed a direct assessment of bacterial growth . The reproducibility of the results and the proven correlation with standard plate count method obtained in industrial conditions make FCM a good predictive method for product and process quality control.

Autoimmunity, 1993, 15(1), 31 - 8
An immunohistochemical study on organized lymphoid cell infiltrates in fetal and neonatal pancreases . A comparison with similar infiltrates found in the pancreas of a diabetic infant; Jansen A et al.; Lymphoid cell infiltrates were analyzed using immunohistochemical techniques on 5 normal fetal and 6 normal neonatal pancreases . Data were compared to data obtained analyzing the lymphoid cell infiltrates in the pancreas of an 8 months old diabetic infant . In the normal fetal and neonatal pancreases islets were intact and not infiltrated . In the diabetic infant beta-cells had vanished in almost all islets, the remaining islets showed a minor infiltration with primarily T-cells, a few B-cells, and some classical macrophages . It appeared that a widespread infiltration of the exocrine pancreas with single dendritic-like cells, and T-cells, and little clusters of these cells were normal features of fetal and neonatal pancreases . In the diabetic case these infiltrative patterns were more pronounced . Larger accumulations of such lymphoid cells could also be detected in the normal fetal and neonatal pancreases and these consisted mainly of T-cell zones, sometimes containing HEV's, with intermingled interdigitating dendritic cells and a few macrophages . This architecture is reminiscent of peripheral lymphoid tissue, such as bronchus-or gut-associated lymphoid tissue . The function of this fetal/neonatal intrapancreatic lymphoid tissue (which disappears in later life) is unknown . Various possibilities are suggested such as a yet unknown ubiquitous fetal/neonatal microbial infection, tolerance induction towards islet cell antigens, an endocrine regulatory function of infiltrated lymphoid cells, and a normal ontogenetic process.

Arch Tierernahr, 1993, 44(3), 283 - 91
Protein degradation in different feedstuffs labelled with 15N by using the nylon bag technique; Katzy R et al.; The nylon bag technique was used to determine the Nitrogen (N) and 15N degradation of 15N labelled feedstuffs in the rumen . The N and 15N degradation values were calculated according to Orskov and McDonald (1979) and ranged from 46.8 to 92.0 and from 61.8 to 93.6%, respectively . The differences between N and 15N degradation values of high fibre content feedstuffs are the highest, thus the measuring errors were greatest here . But differences also existed in concentrates . This study indicated that especially barley had a higher proportion of microbial N in the bag residues after the washing than the other concentrates . Therefore it is necessary to correct the N degradation values not only in cases of high fibre content but also in cases of low nitrogen content of feedstuffs . The calculation of the N degradation values could be possible on the basis of crude fibre and crude protein contents of feedstuffs . But experiments with a much larger number of 15N labelled feedstuffs have to be realized to give an accurate prediction of N degradation.

Rev Argent Microbiol, 1993, 25(1), 15 - 26
{Effect of the vegetative cover on the biological activity of the soil of Chaco Arido}; Abril A et al.; Vegetation plays a primal role in arid ecosystems, since it creates microclimate conditions that moderate the characteristics of the region whereby the rational use of vegetal resources is fundamental . Felling, clearing and overgrazing lead to decrease in organic contribution and stimulate soil compaction, causing an alteration of microbial activity, with losses in nutrient turnover . The global biological activity is a soil parameter easy to obtain and indicates the presence and diversity of soil life as well as substrate availability and is useful in order to characterize soil potential fertility . This work was carried out in Natural Forest Reserve Chancani, Province of Cordoba (Argentina), which is representative of Argentine Dry Chaco . Dominant tree species are: Prosopis flexuosa and Aspidosperma quebracho blanco . The global biological activity (GBA) was measured along one year, under trees, under shrubs and in interspaces . Soil samples were taken monthly from plots with four management systems: 1) forest, ii) selective clearing (only dominant species remain), iii) bush (clearing invaded by Larrea sp) and iv) grazing (cleared area, neither trees nor shrubs) . GBA was evaluated using the CO2 release method, after ten days of incubation . It is concluded that in the plots with grasses and under the trees GBA was higher than with other treatments . The lesser GBA was detected in bushes and interspaces . All differences were more prominent during extreme temperature months . No significant difference between both species of dominant trees was observed.

J Nutr Sci Vitaminol (Tokyo), 1993, 39 Suppl, S23 - 33
Antioxidants in infection; Keusch GT; Endogenous oxidation reactions are essential for the normal biochemistry of life and are especially critical for leukocyte microbial killing mechanisms in host defense to infectious diseases . However, reactive oxidative intermediates can damage normal tissues unless kept under antioxidant control . Three selected examples of oxidant-antioxidant systems involved in infectious diseases are discussed, regulation of molecular iron catalyzed oxidations, superoxide scavengers and inhibitors of nitric oxide synthase in septic shock, and the use of glutathione replacement therapy in HIV infection and AIDS . The data suggest that antioxidants, and therapy based on increasing antioxidant potential, have a major impact on clinical infectious diseases.

Biosystems, 1993, 31(2-3), 135 - 44
Distribution of types of microbial mats at the Ebro Delta, Spain; Guerrero R et al.; The distribution and types of microbial mats of the Ebro Delta (Catalonia, Spain) are described . The studied area is La Banya spit, formed by a narrow sand bar and a peninsula, located south of the main body of the Ebro Delta . Sea water can penetrate into the back shore through channel inlets, cutting the steeper coastal barrier of the open sea side of the spit or through the complex drainage channel system of the low-energy beaches in the inner Alfacs Bay . Sea water can stay in the back shore almost permanently, trapped in isolated depressions and lagoons . The surface distribution of microbial mats in La Banya spit has been studied by means of a detailed interpretation of vertical aerial photographs and field work consisting of mapping and sampling . The observed different ratios of cyanobacteria, as well as the presence and thickness of the layers of anoxygenic sulfur phototrophic bacteria, depend on the moisture content, the system stability, and the age of the microbial mat . Lyngbya, Oscillatoria, and Spirulina are the first cyanobacteria able to colonize the bare sediment . Lyngbya dominates in young microbial mats and in mats exposed to frequent desiccation . Microcoleus is the second most important colonist in the microbial succession . In relation to water, the alternation of emergence and submergence is optimal for the maximal development of Microcoleus-dominated microbial mats . We classify the microbial mats of the Ebro Delta into five main types: (i) Lyngbya-dominated type, in which the anoxygenic phototrophic bacteria are absent and the black layer of sulfate-reducers is very thin; (ii) Spirulina-dominated type; (iii) Oscillatoria-dominated type, which is found only at one site and covers a small area--this type, like the Spirulina-type mat, is not common in the Ebro Delta; (iv) Lyngbya/Microcoleus-transition type, in which Microcoleus and Lyngbya coexist in similar proportions--in the more developed mats of this group a layer of purple bacteria is usually present, and the black layer of sulfate-reducers is usually also well developed; and (v) Microcoleus-dominated type--in La Banya spit, this type is found in localities with relatively stable conditions . These areas are wet during most of the year . After appropriate conditions of diagenesis, the most highly developed microbial mats may be preserved as laminated hard sediments . The field study has been completed with cultivation, isolation, and identification of the main cyanobacterial genera under laboratory conditions.

Biosystems, 1993, 31(2-3), 121 - 5
Origins of species: acquired genomes and individuality; Margulis L; Entire genomes with their accompanying protein synthetic systems are transferred throughout the biosphere primarily as bacteria and protists which become symbionts as they irreversibly integrate into pre-existing organisms to form more complex individuals . Individualization is stabilized by simultaneous transmission of once-separate heterologous genetic systems . The origin of new species is hypothesized to correlate with the acquisition, integration and subsequent inheritance of such acquired microbial genomes . These processes were recognized by Mereschkovsky ("Symbiogenesis" in Russian, 1909) and by Wallin ("Symbionticism", see p . 181, this issue).

J Cell Sci Suppl, 1993, 17, 209 - 15
The role of transepithelial transport by M cells in microbial invasion and host defense; Neutra MR et al.; Transepithelial transport of antigens by M cells in the epithelium associated with lymphoid follicles in the intestine delivers immunogens directly to organized mucosal lymphoid tissues, the inductive sites for mucosal immune responses . We have exploited M cell transport to generate and characterize specific monoclonal IgA antibodies that can prevent interaction of pathogens with epithelial surfaces . The relative protective capacities of specific monoclonal IgA antibodies have been tested in vivo by generation of hybridoma tumors that result in secretion of monoclonal IgA into the intestine . Using this method, we have established that secretion of IgA antibodies recognizing a single surface epitope on enteric pathogens can provide protection against colonization or invasion of the intestinal mucosa.

Reprod Nutr Dev, 1993, 33(6), 531 - 9
Effects of a microbial probiotic (Sporolactobacillus P 44) on postprandial porto-arterial concentration differences of glucose, galactose and amino-nitrogen in the growing pig; Rychen G et al.; Postprandial kinetics of porto-arterial concentration differences of glucose (G), galactose (Gal), L-lactic acid (LA), amino-nitrogen (AN) and urea (U) were studied in the pig after the ingestion of 10(7) colony-forming units (cfu) of Sporolactobacillus P 44 per g of feed . Eight fistulated pigs (portal vein and brachiocephalic trunk; mean body weight 70 +/- 4 kg) were used . The diet was based on skimmed milk (32%), barley (30%), maize (10%) and lactose (7%) . The postprandial blood kinetics, 4 conducted per animal at 1-wk intervals, were studied during the 3 h following the ingestion of test meals of 1,000 g basal diet (BD) or the same diet supplemented by the bacteria (SD) . The apparent absorption was estimated from the area between the portal and arterial concentrations . The areas of porto-arterial differences of G, Gal and AN of SD for the first 3 h after the meal were significantly higher after SD ingestion than those measured after BD intake . Plasma concentrations of U and porto-arterial differences of U and LA were not modified by the probiotic . These effects disappeared immediately after dietary supplement interruption, suggesting that added bacteria presence in the intestinal lumen was fundamental to the modifications observed in apparent absorption.

Eye, 1993, 7 ( Pt 6), 719 - 25
A review of 72 consecutive cases of Acanthamoeba keratitis, 1984-1992; Bacon AS et al.; A review of consecutive cases of Acanthamoeba keratitis presenting since 1984 was undertaken in order to assess prognostic factors, the success of culture procedures and the outcome of medical and surgical management, with reference to current clinical practice . Seventy-two consecutive cases (77 eyes) of Acanthamoeba keratitis have been managed . Sixty-four patients were contact lens wearers, 28 of these wearing disposable lenses . Superficial corneal involvement and perineural infiltrates were common in those diagnosed less than a month after first symptoms, designated 'early' presentation . Ring infiltrates and ulceration with stromal lysis characterised those presenting at 1-2 months ('intermediate') or after 2 months ('late'); these groups also progressed more frequently to hypopyon, scleritis, glaucoma and cataract formation . Positive corneal cultures were obtained in 10 of 14 (71%) intermediate and 17 of 23 (74%) late cases; early cases underwent epithelial biopsy but formal trephine biopsy was not usually justified (1 of 35 cases) and only 19 of 35 (54%) were tissue-positive . Microbial co-isolates were obtained from 20 corneas . Thirty-four penetrating keratoplasties were performed in 23 eyes, 21 whilst inflamed and 13 when quiet . Of 13 failures in inflamed eyes, 9 were due to recurrence of Acanthamoeba infection . Medical cure is known to have been achieved in 64 of 73 (88%) eyes, 4 of the original 77 having been lost to follow-up abroad . Fifty-eight of 73 eyes (79%) achieved a final visual acuity of 6/12, and of the culture-positive cases, 32 of 46 (70%) achieved 6/12.(ABSTRACT TRUNCATED AT 250 WORDS)

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