J Anim Sci, 1993 Apr, 71(4), 1041 - 6 Modeling gas production kinetics of grass silages incubated with buffered ruminal fluid; Beuvink JM et al.; Time course measurements of in vitro gas production of feedstuffs incubated with buffered ruminal fluid give information about the rate at which the feed is fermented . To compare gas production kinetics from different feeds, a mathematical model was needed to describe the data . We investigated several existing models (exponential, logistic, Gompertz, Richards, Schnute), fitting them to 50 gas production curves obtained with grass silages . None of them gave a satisfactory description of the data . A new model was developed (modified Gompertz model; mGom) that basically divided gas production into two fractions, one arising from rapidly fermentable feed components and the other from slowly fermentable feed components . Residual mean squares (RMS) for the mGom model were lower (P < .05; n = 50) than the RMS for the other models . A good statistical comparison of kinetic gas production data is made possible with the mGom model. Am J Respir Cell Mol Biol, 1993 Apr, 8(4), 358 - 64 Enhanced secretion of immune-modulating cytokines by human lung fibroblasts during in vitro infection with Mycoplasma fermentans; Fabisiak JP et al.; Fibroblasts may play an important role in the modulation of immune and inflammatory responses through elaboration of cytokines . To test this hypothesis, human lung fibroblasts were isolated from transbronchial biopsy specimens and assayed for production of interleukin-6 (IL-6) and granulocyte/macrophage colony-stimulating factor (GM-CSF) . The sources of fibroblasts included lung allografts, recipient lungs obtained at time of transplant, and normal lung tissue removed during tumor resection . During the course of these studies, several early-passage fibroblasts from transplant recipients were observed to contain mycoplasma (MP)-like organisms as detected by extranuclear fluorescent staining with Hoechst 33258 . Positive staining cultures were associated with isolation of Mycoplasma fermentans . IL-6 and GM-GSF as measured by ELISA were found to be elevated over 50-fold in conditioned medium from MP-infected fibroblasts as compared with noninfected lines . Treatment of cells with mycoplasma removal agent (MRA) eliminated extranuclear Hoechst fluorescence and significantly reduced the production of these cytokines . Tumor necrosis factor-beta (TNF-beta) induction of IL-6 and GM-CSF was amplified synergistically in infected cultures . No additional production of IL-6 or GM-CSF was observed in infected cultures treated with interferon-gamma (IFN-gamma) despite the ability of IFN-gamma to modestly induce IL-6 in uninfected cultures . Thus, in vitro infection of lung fibroblasts with MP represents a potent stimulus for the production of inflammatory cytokines and, therefore, necessitates rigorous control for these organisms in cell culture studies.(ABSTRACT TRUNCATED AT 250 WORDS) J Bacteriol, 1993 Apr, 175(8), 2205 - 13 Energy flux and osmoregulation of Saccharomyces cerevisiae grown in chemostats under NaCl stress; Olz R et al.; The energetics and accumulation of solutes in Saccharomyces cerevisiae were investigated for cells grown aerobically in a chemostat under NaCl stress and glucose limitation . Changed energy requirements in relation to external salinity were examined by energy balance determinations performed by substrate and product analyses, with the latter including heat measurements by microcalorimetry . In both 0 and 0.9 M NaCl cultures, the catabolism was entirely respiratory at the lowest dilution rates tested but shifted to a mixed respiratory-fermentative metabolism at higher dilution rates . This shift occurred at a considerably lower dilution rate for salt-grown cells . The intracellular solute concentrations, as calculated on the basis of intracellular soluble space determinations, showed that the internal Na+ concentration increased from about 0.02 molal in basal medium to about 0.18 molal in 0.9 M NaCl medium, while intracellular K+ was maintained around 0.29 molal despite the variation in external salinity . The intracellular glycerol concentration increased from below 0.05 molal at low salinity to about 1.2 molal at 0.9 M NaCl . The concentrations of the internal solutes, however, changed insignificantly with growth rate and energy metabolism . The additional maintenance energy expenditure for growth at 0.9 M NaCl was, depending on the growth rate, 14 to 31% of the total energy requirement for growth at 0 M NaCl . Including the energy conserved in glycerol, the total additional energy demand for growth at 0.9 M NaCl corresponded to 28 to 51% of the energy required for growth at 0 M NaCl. J Nutr, 1993 Apr, 123(4), 681 - 8 Interaction between methane-producing status and diet on serum acetate concentration in humans; Wolever TM et al.; About half the population excretes methane in the breath . To see if methane producing status influenced serum acetate, we studied six methane producers and six nonproducers on three separate days . For 36 h they ate a polysaccharide-free diet alone, or with 20 g of unabsorbed sugar lactulose, or 20 g of fermentable fiber, guar, in random order . The mean fasting serum acetate concentration on the three test days in producers was higher than in nonproducers, 84 +/- 5 vs . 69 +/- 5 mumol/L (P < 0.05) . Compared to the control diet, both lactulose and guar raised serum acetate concentration significantly in both groups of subjects . However, there was a significant interaction between methane producing status and diet . After lactulose consumption, postprandial serum acetate was similar in both groups of subjects, but guar consumption had a significantly greater effect in producers than nonproducers (98 +/- 8 vs . 73 +/- 5 mumol/L; P < 0.05) . We conclude that methane producing status may influence serum acetate concentrations in humans, depending upon the type of fermentable carbohydrate in the diet. J Nutr, 1993 Apr, 123(4), 676 - 80 Fate of beta-cyclodextrin in the human intestine; Flourie B et al.; We assessed the fate of beta-cyclodextrin, which is composed of seven alpha(1-->4)-linked glucose units in ring structure, in the human gastrointestinal tract . In four healthy ileostomists, ileal effluent was collected after oral administration of beta-cyclodextrin during fasting (10 g of beta-cyclodextrin) and postprandially (10 g of beta-cyclodextrin three times daily with meals) . In 10 healthy volunteers, the amount of beta-cyclodextrin passing into the colon was determined by means of the breath hydrogen technique using lactulose as a standard, and stools were collected after oral administration of beta-cyclodextrin during fasting (10 g of beta-cyclodextrin) and postprandially (10 g of beta-cyclodextrin three times daily with meals) . In ileostomists, we recovered from the small intestine 91 +/- 5% and 97 +/- 10% (mean +/- SD) of beta-cyclodextrin ingested during fasting and with meals, respectively . In healthy volunteers, H2 excretion in breath after beta-cyclodextrin ingestion was low compared with excretion after lactulose, but only traces of beta-cyclodextrin were recovered in stools . We conclude that beta-cyclodextrin is poorly hydrolyzed in the human small intestine but that it is fermented by the colonic flora with apparent minimal H2 production. J Dairy Sci, 1993 Apr, 76(4), 1091 - 105 Nonstructural carbohydrate and protein effects on rumen fermentation, nutrient flow, and performance of dairy cows; Aldrich JM et al.; Four multiparous Holstein cows fitted with rumen and duodenal cannulas were used in a 4 x 4 Latin square with 20-d periods . Four diets were formulated for high and low rumen availabilities of nonstructural carbohydrate and protein . Cows were milked and fed three times daily . Milk production averaged 39 kg/d and was unaffected by treatment . Dry matter intake and 4% FCM production were increased by 1 kg/d for cows fed the low rumen-available nonstructural carbohydrate diets . Milk protein percentage was elevated when either the high rumen-available nonstructural carbohydrate or high rumen-available protein diets were fed . Low rumen-available protein diets increased duodenal passage of total essential AA but did not increase passage of Arg, Ile, and Met . Passage of bacterial N was highest (262 g/d) when high rumen-available nonstructural carbohydrate was combined with high rumen-available protein and was lowest when high rumen-available nonstructural carbohydrate was combined with low rumen-available protein (214 g/d) . Diets with low rumen availabilities of protein were not advantageous, possibly because duodenal passage of one or more of the limiting AA was not increased . Passage of bacterial N to the duodenum was highest when rumen availabilities of both nonstructural carbohydrate and protein were high. J Anim Sci, 1993 Apr, 71(4), 1032 - 40 Effect of nonfiber carbohydrate level and Aspergillus oryzae fermentation extract on intake, digestion, and milk production in lactating dairy cows; Sievert SJ et al.; Eight multiparous, ruminally cannulated Holstein cows averaging 40 d in milk and 575 kg BW at the start of trial were in a replicated 4 x 4 Latin square arrangement (28-d periods) to determine the effects of dietary nonfiber carbohydrate (NFC) level and Aspergillus oryzae fermentation extract (AO) on intake, milk production, and nutrient digestibility . Treatments were 42 or 35% NFC and 0 or 3 g of AO per day arranged as a 2 x 2 factorial . Diets formulated to contain 21% NDF from alfalfa silage (48.4% of ration DM) and 18.5% CP were fed as total mixed rations twice daily . Alteration of dietary NFC level was by partial replacement of shelled corn and soybean meal with wheat middlings and brewers dried grains . Intake of NDF was higher (1.49 vs 1.22% of BW) for 35 than for 42% NFC diets, but DMI was lower (24.1 vs 24.9 kg/d) . Milkfat percentage, ruminal pH, ammonia, acetate (moles/100 moles), and total tract digestibility of fiber were higher for 35% NFC diets; however, ruminal disappearance of DM, CP, and NDF from Dacron bags containing alfalfa hay was not affected (P > .10) by NFC level . Supplementation with AO did not affect (P > .10) DMI, milk yield, or nutrient digestion . Partial replacement of corn with high-fiber byproducts to lower dietary NFC level and correspondingly increase NDF level increased NDF intake but effected only a small change in DMI . Reducing dietary NFC level improved ruminal fermentation and milkfat percentage without significantly affecting milk yield. Proc Natl Sci Counc Repub China B, 1993 Apr, 17(2), 62 - 9 Recovery and properties of a fructooligosaccharides-producing beta-fructofuranosidase from Aspergillus japonicus CCRC 38011; Su YC et al.; The fungus (Aspergillus japonicus CCRC 38011) was found to be able to produce beta-fructofuranosidase with high transfructosylating activity (Ut), a key enzyme involved in synthesis of fructooligosaccharides from sucrose . The Ut productivities of this microorganism were 191.5 units/ml broth in a 6-L jar-fermentor and 256.1 units/ml broth in a 1000-L pilot-fermentor in a modified medium containing 8% sucrose as a carbon source . Most of the Ut of this microorganism was found to be bound with mycelia . Incubation and sonication treatment extracted this enzyme from mycelia with maximum efficiencies of 66.3% and 44.3%, respectively . On the other hand, homogenization and freeze-thawing treatment had only a small effect on enzyme extraction . The soluble enzyme extracted from mycelia by incubation at pH 5.0 and 40 degrees C for 3 hours could be easily recovered and purified by acetone precipitation . The recovery of Ut from the crude enzyme solution was 99.5% by mixing the solution with an equal volume of acetone at 4 degrees C, followed by centrifugation . The purification factor of acetone precipitation was 15.8 . The optimum pH and temperature of Ut was 5.0 and 65-70 degrees C, respectively . The enzyme was stable at a pH between 4.0 and 5.0 and at a temperature below 60 degrees C. Curr Opin Biotechnol, 1993 Apr, 4(2), 183 - 7 Biosensors for fermentation control; Pons MN; Over the past year, biosensor development has been an active area of research . The actual application of biosensors in process monitoring and control is limited to a few cases, mainly as a result of difficulties relating to their long-range stability and their sensitivity to interfering compounds . Steam sterilization is no longer a problem though, as the great majority of sensors are part of flow-injection analysis systems. Appl Microbiol Biotechnol, 1993 Apr, 39(1), 21 - 5 Improvement of performance for cross-flow membrane filtration of pullulan broth; Yamasaki H et al.; To improve the performance of cross-flow membrane filtration of pullulan broth from Aureobasidium pullulans, the effect of the cultivation conditions was examined . In particular, the sucrose concentration in the medium was changed over a wide range . By decreasing the sucrose concentration the distribution of morphology of the microbial cells in the broth changed; the yeast-like form became predominant and, as a result, the specific resistance of the microbial cake was lowered . When the broth was fermented with a sucrose concentration of 2.5% or lower, the filtration characteristics were greatly improved by periodic closure of permeation during cross-flow filtration. Fiziol Zh Im I M Sechenova, 1993 Apr, 79(4), 61 - 7 {The role of the proteins of the blood plasma in the homeostasis of its amylase and pepsinogen}; Korot'ko GF et al.; Hypervolemia and stimulation of diuresis in dogs was found to decrease the amylolytic activity of the blood plasma and pepsinogen concentration . The number of enzymes bound with blood plasma albumins decreases in hyper-fermentation . The changing connexion of the plasma albumins with the enzymes entails a renal (as well as extrarenal) extraction of the enzymes from the organism and maintenance of their relatively constant concentration and activity in the blood. J Antibiot (Tokyo), 1993 Apr, 46(4), 545 - 53 Fiscalins: new substance P inhibitors produced by the fungus Neosartorya fischeri . Taxonomy, fermentation, structures, and biological properties; Wong SM et al.; Three new compounds, named fiscalins A, B, and C, were found in culture broth produced by a Neosartorya fischeri . These compounds inhibit the binding of radiolabeled substance P ligand to the human neurokinin (NK-1) receptor, with Ki values of 57, 174, and 68 microM, respectively . Detailed spectroscopic and amino acid analyses led to the elucidation of structures for the three fiscalins . The structures contain an indolyl moiety linked to an athranilic acid derived tricyclic system . The absolute configuration of fiscalin A was determined by X-ray crystallography and chiral amino acid analysis . The presence of fiscalins was detected directly in crude cellular extracts using LC-MS methods. Int J Syst Bacteriol, 1993 Apr, 43(2), 293 - 6 Proposal of Quinella ovalis gen . nov., sp . nov., based on phylogenetic analysis; Krumholz LR et al.; Quin's oval is a relatively large bacterium often seen in the rumens of sheep fed diets containing some readily fermented carbohydrates . It has not been obtained in axenic cultures, but a number of its features have been determined by various methods, such as studying cell suspensions purified from rumen fluid by differential centrifugation . We obtained similarly purified suspensions from a sheep fed a diet containing a large amount of molasses . Nearly complete 16S rRNA sequence analysis of these cells as well as cells as Selenomonas ruminantium subsp . ruminantium GA192 (ATCC 12561; type strain) and S . ruminantium subsp . lactilytica HD4 (ATCC 27209) was done . These sequences were compared with those of other bacteria . Evolutionary distance estimates indicated that Quin's oval was most closely related to the Selenomonas-Megasphaera-Sporomusa group in the gram-positive phylum but that it belongs in a new genus . We propose the name Quinella ovalis gen . nov., sp . nov., with its description based on previously known features. FEMS Microbiol Lett, 1993 Mar 15, 108(1), 47 - 52 Arginine utilization by Mycoplasma fermentans is not regulated by glucose metabolism: a 13C-NMR study; Olson LD et al.; 13C-NMR studies on the effect of glucose metabolism on arginine hydrolysis in Mycoplasma fermentans cells have been performed using a continuous perfusion technique . With this procedure we were able to show, in the presence of glucose, the rapid accumulation of lactic acid and, in the presence of arginine, the formation of citrulline that is apparently further metabolized . As the accumulation of lactate and the breakdown of arginine were observed in the simultaneous presence of both substrates, it is suggested that the glucose utilization has little or no effect on the deimination of arginine to citrulline. Biochem J, 1993 Mar 15, 290 ( Pt 3), 723 - 9 Expression, purification and characterization of B72.3 Fv fragments; King DJ et al.; The Fv fragment of the antibody B72.3 has been produced by expression in both a mammalian and microbial system, namely Chinese hamster ovary (CHO) cells and Escherichia coli . In both cases secretion of the Fv into the culture medium was achieved, with equivalent amounts of Vh and Vl produced . The yield of Fv from CHO cells was 4 mg/l in roller-bottle culture . E . coli proved to be a more productive system with yields of 40 mg/l in shake flasks rising to 450 mg/l in fermentations . B72.3 Fv from both sources was capable of binding to antigen with similar binding ability to the Fab' fragment . A detailed sedimentation analysis, both by velocity and equilibrium techniques, revealed that the two domains of Fv are associated at high concentrations at pH values close to neutral, but dissociate at concentrations lower than approx . 0.5 mg/ml . Individual Vh or Vl polypeptides are not able to bind to the antigen and thus these results suggest that the antigen promotes assembly of Fv at the low concentrations used in the antigen-binding assays . At a pH value of 1.9, Vh and Vl are completely dissociated even at very high concentrations and are apparently unfolded at low solute concentrations . Small-angle X-ray scattering was used to measure a radius of gyration of 1.75 +/- 0.2 nm (17.5 +/- 2 A) for Fv. FEBS Lett, 1993 Mar 8, 318(3), 345 - 52 Purification and characterization of the trefoil peptide human spasmolytic polypeptide (hSP) produced in yeast; Thim L et al.; Recombinant human spasmolytic polypeptide (r-hSP) has been produced in relatively large amounts in Saccharomyces cerevisiae . The two intronless trefoil domains of the hSP-DNA were cloned separately by PCR from human genomic DNA, and the remaining parts of the gene synthesized . Recombinant plasmids were constructed to encode a fusion protein consisting of a hybrid leader sequence and the hSP sequence . The leader sequence serves to direct the fusion protein into the secretory pathway of the cell and to expose it to the Kex 2 processing enzyme system . The secreted r-hSP was found in a glycosylated and an non-glycosylated form . The two forms of r-hSP were purified from the yeast fermentation broth by a combination of ion-exchange chromatography and preparative HPLC . The overall yield from 8 litres of fermentation broth was 160 mg r-hSP and 219 mg glycosylated r-hSP corresponding to 50% and 34%, respectively . The structure of the r-hSP and the glycosylated r-hSP was determined by amino acid analysis and carbohydrate composition analysis as well as by peptide mapping, amino acid sequencing and mass spectrometric analysis. FEMS Immunol Med Microbiol, 1993 Mar, 6(2-3), 207 - 12 Porphyromonas-like gram-negative rods in naturally occurring periodontitis in dogs; Karjalainen J et al.; A total of 259 Gram-negative Porphyromonas-like rods isolated from subgingival plaque samples of 16 family-owned dogs with naturally occurring periodontitis were characterized phenotypically by biochemical reactions, metabolic end products and enzymatic activities (API-ZYMTM, RoscoTM) . Four distinct groups were found . Group A isolates (63) were asaccharolytic, lipase negative, trypsin positive and produced phenylacetic acid (PAA) from peptone-yeast extract glucose broth . Unlike P . gingivalis strains they were catalase positive . Group B isolates (42) differed from those of group A by a positive lipase reaction and from those of group D by failing to ferment sugars . Group C isolates (88) were asaccharolytic and did not produce PAA . They were alpha-fucosidase, N-acetyl-beta-glucosaminidase (beta-NAG) and trypsin negative, resembling P . endodontalis, but unlike human isolates, they were catalase positive . Subgroup C.1 isolates (6) differed from those of parent group C by producing minor amounts of PAA, and subgroup C.2 isolates (12) were beta-NAG positive . Group D isolates (46) were weakly fermentative, lipase, catalase and trypsin positive, and produced PAA . They resembled the B (P.) salivosus type strain which, in our hands, fermented weakly glucose, lactose and mannose . Two isolates could not be assigned to any of the previous groups. Br J Nutr, 1993 Mar, 69(2), 511 - 25 Assessment of fermentation in growing pigs given unmolassed sugar-beet pulp: a stoichiometric approach; Zhu JQ et al.; In four experiments growing pigs were given a cereal-based diet alone or supplemented with unmolassed sugar-beet pulp (SBP), used as a model substrate for fermentation . The rates of production of methane and gaseous hydrogen were measured and, together with the molar proportions of volatile fatty acids (VFA) in the digesta, used in stoichiometric calculations of fermentation . The resulting estimates were only one-sixth of the observed extent of digestion of SBP . Bacteriostatic levels of antibiotics reduced fermentation by more than half, as judged from the digestion of non-starch polysaccharides: allowing for the incomplete suppression of fermentation it was estimated that the production of methane and VFA could account completely for the digested SBP . The potential contribution of various routes of hydrogen disposal to the error of the stoichiometric calculations is discussed. Br J Nutr, 1993 Mar, 69(2), 497 - 509 Complex carbohydrate digestion and large bowel fermentation in rats given wholemeal bread and cooked haricot beans (Phaseolus vulgaris) fed in mixed diets; Key FB et al.; The digestion of non-starch polysaccharides (NSP) and of resistant starch (RS) by rats fed on wholemeal-bread-based diets containing 0-450 g cooked, freeze-dried haricot beans (Phaseolus vulgaris)/kg diet was measured over the final 14 d of a 21 d feeding experiment . The bread and beans provided all the dietary polysaccharide . RS could not be detected consistently in faeces and it was assumed that this fraction was entirely fermented in the large bowel (LB) . NSP digestibilities were 0.56 and 0.86 for wholemeal bread and beans respectively with no evidence that the dietary presence of beans affected digestibility of bread NSP . Bean non-cellulosic polysaccharides were highly digestible with values of 0.98, 0.88 and 0.99 for arabinose, xylose and uronic acids components respectively . There were large increases in organic matter flow to the LB when beans were fed which was associated with marked caecal hypertrophy and alterations in caecal volatile fatty acids (VFA) pattern . Calculated VFA absorption from the LB was 5-fold higher with the highest level of beans and this was reflected in higher concentrations of VFA in portal and heart blood. Br J Nutr, 1993 Mar, 69(2), 481 - 95 Gastrointestinal responses of rats fed on white and wholemeal breads: complex carbohydrate digestibility and the influence of dietary fat content; Key FB et al.; To obtain quantitative information on the digestibility of the non-starch polysaccharides (NSP) fraction of white and wholemeal breads, rats were fed on diets in which freeze-dried bread (white, wholemeal or mixtures of the two) provided all the complex carbohydrates . In a second experiment the possibility that dietary fat concentration might influence NSP digestibility was tested by feeding diets containing 30 or 170 g maize oil/kg and either white or wholemeal bread . Multiple linear regression analysis provided little evidence of associative effects of dietary components on NSP digestibility and in the two experiments digestibilities of NSP for white and wholemeal breads were 0.77-0.82 and 0.47-0.52 respectively . Xylose- and arabinose-containing polymers were better digested than was cellulose for both breads . Replacing white by wholemeal bread markedly increased the molar proportion of butyrate in caecal volatile fatty acids at the expense of acetate . This was associated with greater flows of organic matter to the large bowel (LB) and a reduction in caecal transit time (Expt 2) . There was little detectable effect of dietary maize oil concentration on NSP digestibility or on LB fermentation . All breads contained some starch resistant to pancreatic alpha-amylase (EC 3.2.1.1) without previous treatment with dimethyl sulphoxide . The digestibility of this starch fraction was not significantly different from 1.0 for all diets except that containing wholemeal bread and the higher maize oil concentration where the apparent digestibility was 0.89. Metabolism, 1993 Mar, 42(3), 347 - 52 Effects of colonic fermentation on respiratory gas exchanges following a glucose load in man; Ritz P et al.; Colonic fermentation produces short-chain fatty acids (SCFA) . In humans, the amount of energy produced from the oxidation of these compounds is unknown and could modify the metabolic utilization of energetic fuels (eg, carbohydrates and lipids) . If it were so, the equations used to evaluate the oxidation of nutrients from indirect calorimetry data should include the contribution of SCFA, which is not usually the case . Indeed, this fermentation process is usually considered as a minor and neglected energetic pathway . In this study, we have addressed the reliability of this assumption . Six normal subjects received orally either 50 g glucose or 50 g glucose plus 20 g lactulose . Their respiratory gas exchanges, breath hydrogen, methane, and 13CO2 concentrations, and plasma glucose, insulin, and free fatty acid (FFA) concentrations were monitored for 8 hours . CO2 production and breath hydrogen concentration were significantly greater with lactulose . No differences in oxygen consumption, breath 13CO2 production, or plasma concentrations of blood glucose, FFA, and insulin could be found between the two experiments . This suggests that the fermentation process induced by lactulose generates extra fuels going through an oxidation pathway . Therefore, the classic equations used to calculate carbohydrate and lipid oxidation and energy expenditure (EE) from indirect calorimetry data are probably not valid when fermentation is taking place . Indeed, in this experiment we could have overestimated glucose oxidation (12.5%) if the fermentation process were not considered . In conclusion, colonic fermentation in humans of nondigestible carbohydrates produces energetic substrates that could be used and oxidized as energetic fuels.(ABSTRACT TRUNCATED AT 250 WORDS) Appl Environ Microbiol, 1993 Mar, 59(3), 786 - 90 Evaluation of colilert-marine water for detection of total coliforms and Escherichia coli in the marine environment; Palmer CJ et al.; A test that allows for early detection of fecally contaminated coastal water would enhance public health protection . Colilert-Marine Water (Colilert-MW; Environetics, Branford, Conn.) is a rapid 24-h test that has recently been developed to detect total coliforms and Escherichia coli in coastal water . We performed a premarketing evaluation of the Colilert-MW product, testing it in parallel with the multiple tube fermentation (MTF) method for 86 coastal water samples in southern California . Statistical analysis was performed by using paired t tests and linear regression . Bacterial isolates were evaluated by biochemical and genetic analysis . The results of this study showed a strong correlation between the traditional MTF and the Colilert-MW method for detection of total coliforms (r = 0.95) and E . coli (r = 0.89) in ocean water samples . Paired t-test results indicated that the Colilert-MW and MTF were equivalent in detecting E . coli and that the Colilert-MW may be more sensitive in the detection of total coliforms . We conclude that Colilert-MW would be a useful tool with which to monitor coastal beach water. Appl Environ Microbiol, 1993 Mar, 59(3), 748 - 55 Glucose and carbon dioxide metabolism by Succinivibrio dextrinosolvens; O'Herrin SM et al.; Growth rates and culture conditions affect the molar yields of catabolic end products and cells of Succinivibrio dextrinosolvens growing on glucose . When growth in chemostats occurred, a trend toward decreased succinate and acetate formation, increased lactate formation, and a higher yield of cells correlated with an increase in the growth rate . End product and cellular yields on defined medium indicate a high maintenance requirement for S . dextrinosolvens and are consistent with energy conservation steps during the formation of acetate and succinate . Simultaneous carbon dioxide consumption and production were determined from batch studies with NaH14CO3, and the amounts were used to calculate a fermentation balance . These data also indicated that CO2 consumption lags behind CO2 production early in the growth phase, becoming equivalent to it toward stationary phase . Significantly more CO2 was fixed by S . dextrinosolvens when the organism was cultured in chemostats sparged with CO2 . Formate is in part derived from free CO2 in the medium, as shown by 13C nuclear magnetic resonance studies, and may be sensitive to CO2 availability . Nuclear magnetic resonance data are consistent with the carboxylation of a C3 intermediate of the Embden-Meyerhof-Parnas pathway of glycolysis to a C4 compound to eventually form succinate. Appl Environ Microbiol, 1993 Mar, 59(3), 729 - 33 Production of high concentrations of ethanol from inulin by simultaneous saccharification and fermentation using Aspergillus niger and Saccharomyces cerevisiae; Ohta K et al.; Pure nonhydrolyzed inulin was directly converted to ethanol in a simultaneous saccharification and fermentation process . An inulinase-hyperproducing mutant, Aspergillus niger 817, was grown in a submerged culture at 30 degrees C for 5 days . The inulin-digestive liquid culture (150 ml) was supplemented with 45 g of inulin, 0.45 g of (NH4)2SO4, and 0.15 g of KH2PO4 . The medium (pH 5.0) was inoculated with an ethanol-tolerant strain, Saccharomyces cerevisiae 1200, and fermentation was conducted at 30 degrees C . An additional 20 g of inulin was added to the culture after 15 h of fermentation . S . cerevisiae 1200 utilized 99% of the 65 g of inulin during the fermentation, and produced 20.4 and 21.0% (vol/vol) ethanol from chicory and dahlia inulins, respectively, within 3 days of fermentation . The maximum volumetric productivities of ethanol were 6.2 and 6.0 g/liter/h for chicory and dahlia inulins, respectively . The conversion efficiency of inulin to ethanol was 83 to 84% of the theoretical ethanol yield. J Antibiot (Tokyo), 1993 Mar, 46(3), 420 - 9 Biosynthesis of the pradimicin family of antibiotics . II . Fermentation, isolation and structure determination of metabolites associated with the pradimicins biosynthesis; Tsuno T et al.; Ten metabolites produced by 4 mutants derived from Actinomadura verrucosospora subsp . neohibisca E-40, a high pradimicins producer, were isolated and their structures were determined . Strain JN-219 produced 3 novel analogs of the pradimicin A aglycone, i.e . 11-O-demethyl-7-methoxypradinone II and 11-O-demethylpradinones I and II together with a known aglycone analog, pradinone I, while the metabolites from strain JN-47 were determined to be 2 new aglycone analogs, 11-O-demethylpradimicinone I and 11-O-demethyl-7-methoxypradimicinone II and a known aglycone analog, 11-O-demethylpradimicinone II (11dM-PMN II) . Products of strain JN-207 were identified as 11-O-demethyl-6-deoxypradinone I and 11dM-PMN II . Interestingly, a new pradimicin analog, 7-hydroxypradimicin A was isolated from strain JN-58 together with a new aglycone analog, pradimicinone II and 11dM-PMN II . None of these metabolites showed antifungal activity. J Antibiot (Tokyo), 1993 Mar, 46(3), 412 - 9 Biosynthesis of the pradimicin family of antibiotics . I . Generation and selection of pradimicin-nonproducing mutants; Furumai T et al.; Germinated spores of Actinomadura verrucosospora subsp . neohibisca E-40, a high pradimicins producer, were mutagenized by N-methyl-N'-nitro-N-nitrosoguanidine and/or UV treatment . Thirty-seven mutants which did not produce pradimicin were selected to test for cosynthesis ability, and classified into nine classes . On the basis of their cosynthesis ability and bioconversion results, we concluded that strain JN-213 (class III) was a true converter and that strains JN-219 (class IV), JN-47 (class V) and JNU-46 (class VI) were secretors accumulating biosynthetic intermediates of pradimicin, and that strains JN-59 (class VII), JN-58 (class VIII) and JN-207 (class IX) were producers of shunt metabolites of pradimicin biosynthesis . TLC and HPLC analyses of the fermentation broths of individual strains showed that 8 new compounds were produced along with pradinone I, pradimicinone I, 11-O-demethylpradimicinone II and 7-O-methylpradimicinone II. J Antibiot (Tokyo), 1993 Mar, 46(3), 380 - 6 5-N-acetylardeemin, a novel heterocyclic compound which reverses multiple drug resistance in tumor cells . II . Isolation and elucidation of the structure of 5-N-acetylardeemin and two congeners; Hochlowski JE et al.; A family of novel compounds has been detected and isolated following an assay for the attenuation of multiple drug resistance in tumor cells from the fermentation broth and mycelia of a strain of Aspergillus fischeri which we have designated var . brasiliensis . The structures of three components were determined employing 1-D and 2-D homonuclear and heteronuclear NMR spectroscopy and mass spectrometry . The structure of 5-N-acetylardeemin was confirmed by single crystal X-ray diffraction . These compounds are most closely structurally related to asperlicin E1). J Antibiot (Tokyo), 1993 Mar, 46(3), 374 - 9 5-N-acetylardeemin, a novel heterocyclic compound which reverses multiple drug resistance in tumor cells . I . Taxonomy and fermentation of the producing organism and biological activity; Karwowski JP et al.; The ardeemins are a new family of secondary metabolites produced by submerged fermentation of a fungus which was isolated from a soil sample collected in Brazil . Based on taxonomic studies, the producing culture was identified as Aspergillus fischeri var . brasiliensis strain AB 1826M-35 . 5-N-Acetylardeemin potentiated the cytotoxicity of the anticancer agent vinblastine in multidrug resistant human tumor cells. Chem Pharm Bull (Tokyo), 1993 Mar, 41(3), 557 - 60 Two new steroidal saponins from dried fermented residues of leaf-juices of Agave sisalana forma Dong No . 1; Ding Y et al.; In a previous paper, we reported the isolation and structure determination of three new steroidal saponins, dongnosides C (3), D (2) and E (1) from the dried fermented residues of leaf-juices of Agave sisalana forma Dong No . 1 . In a continuing study on this plant, two additional new major steroidal saponins, named dongnosides B (4) and A (5), were obtained . Their structures were characterized respectively as tigogenin 3-O-alpha-L-rhamonpyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->2)- {beta-D- glucopyranosyl-(1-->3)}-beta-D-glucopyranosyl-(1-->4)-beta-D-galactop yranoside and 3-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->2)-{beta- D- xylopyranosyl-(1-->3)-beta-D-glucopyranosyl-(1-->3)}-beta-D- glucopyranosyl-(1-->4)-beta-D-galactopyranoside on the basis of chemical and physicochemical evidence. J Dairy Sci, 1993 Mar, 76(3), 826 - 30 Another theory for the action of ruminal buffer salts: decreased starch fermentation and propionate production; Russell JB et al.; Sodium carbonates have been fed to ruminants for more than 20 yr and, in many cases, have alleviated milk fat depression . These effects usually have been ascribed to increased ruminal buffering capacity, but this mode of action has several problems . For the buffering capacity to increase, the concentrations of ruminal bicarbonate, dissolved CO2, and Na have to increase . Ruminal fluid already is saturated with CO2, and the cation concentration of ruminal fluid is regulated closely to prevent hemoconcentration or hemodilution . Based on these latter observations, a significant increase in ruminal buffering capacity is unlikely . The action of bicarbonates is explained more easily by increased water intake, increased ruminal fluid dilution rate, increased flow of undegraded starch from the rumen, and decreased ruminal propionate production. J Dairy Sci, 1993 Mar, 76(3), 775 - 89 Diets containing high oil corn and tallow for dairy cows during early lactation; Elliott JP et al.; Four multiparous Holstein cows averaging 49 DIM and fitted with ruminal cannulas were utilized in a 4 x 4 Latin square design with 21-d periods to determine the effects of diets containing high oil corn grain and tallow . Cows were fed diets of alfalfa haylage and concentrate (37:63, DM basis) for ad libitum intake . Treatments were 1) control, no added fat; 2) high oil corn grain replacing regular corn grain; 3) high oil corn grain and 2.5% tallow; and 4) high oil corn grain and 5% tallow . Intake of DM, milk production, and yields of milk fat, milk protein, milk SNF, and 4% FCM were not affected by dietary fat, although DMI tended to be lower when cows were fed 5% tallow . Percentages of protein and SNF in milk were lower when cows were fed diets containing fat . Percentage of milk fat was lower when cows were fed diets containing tallow . Supplemental fat decreased total VFA concentrations in ruminal fluid . Cows fed high oil corn had a greater molar proportion of acetate and a larger acetate to propionate ratio in ruminal fluid than cows fed high oil corn and tallow . Digestibility of total fatty acids decreased when tallow was added to diets containing high oil corn . No differences were observed among treatments for total tract apparent digestibilities of DM and fiber or utilization of energy and N . Dietary high oil corn and 5% tallow tended to decrease DMI and to alter ruminal fermentation characteristics. Clin Exp Metastasis, 1993 Mar, 11(2), 201 - 12 U-77,863: a novel cinnanamide isolated from Streptomyces griseoluteus that inhibits cancer invasion and metastasis; Welch DR et al.; Several cinnamoyl compounds have been shown to have antitumor activities, but not specifically anti-invasive or antimetastatic effects . U-77,863 (o-methyl cinnanamide) was originally isolated from a fermentation beer of Streptomyces griseoluteus and recently synthesized (Harper, DE and Welch DR . Journal of Antibiotics, in press) . Based upon some differential activities of cinnanamides, in general, and U-77,863, specifically, we tested the hypothesis that U-77,863 could inhibit invasion and metastasis of human malignant melanoma cell lines C8161 and A375M . Pretreatment of melanoma cells in vitro with nontoxic doses of U-77,863 caused a dose-, and time-dependent, reversible reduction (IC50 = 12.5 micrograms/ml) of invasion through Matrigel-coated polycarbonate filters in the Membrane Invasion Culture System (MICS) . Likewise, lung colonization was significantly (P < 0.05) inhibited when tumor cells were pretreated in vitro with U-77,863 prior to intravenous injection . Structure-activity analysis revealed that the acrylamide side-chain alone and cinnanamide were only slightly less potent than U-77,863, whereas cinnamic acid analogs did not inhibit tumor cell invasion at doses < or = 100 micrograms/ml . U-77,863 inhibits invasion and metastasis without decreasing growth rates or clonogenic potential . Adhesion to endothelial monolayers or extracellular matrices (Matrigel) is not affected by exposure to U-77,863 . U-77,863 presumably inhibits metastasis by inhibiting tumor cell extravasation (invasion) . U-77,863 is a lead compound for developing a novel class of anti-invasive/anti-metastatic drugs. Z Gastroenterol, 1993 Mar, 31(3), 179 - 82 The application of 13C-labelled short chain fatty acids to measure acetate and propionate production rates in the large intestines . Studies in a pig model; Breves G et al.; The production rates of acetate and propionate were measured in the large intestine of pigs by applying the single injection technique of (1-13C)acetate and (1-13C)propionate . Both acids were injected individually through the caecal cannula and for both acids the experiments were performed during two diets with different crude fibre contents . For acetate the increase of dietary crude fibre from 5.1 to 18.3% of dry matter resulted in an increase of mean production rate from 27.4 to 56.2 mmol/h . The mean propionate production rate was raised from 3.6 to 7.0 mmol/h when the dietary crude fibre was increased from 4.4 to 24.3% . From both experimental series the contribution of hindgut fermentation to energy maintenance requirement were estimated to be in a range between 7 and 40% depending on the body weight of the animals and the percentage of dietary crude fibre. Gut, 1993 Mar, 34(3), 386 - 91 Butyrate production from dietary fibre and protection against large bowel cancer in a rat model; McIntyre A et al.; Butyrate slows the growth of cancer cells cultured in vitro . To determine the relevance of the fermentative production of butyrate in vivo, colonic butyrate concentrations were manipulated by feeding different dietary fibres and were related to tumour development in the rat dimethylhydrazine model of large bowel cancer . It has previously been shown that guar gum and oat bran, while highly fermentable, are associated with low butyrate levels in the distal colon, while wheat bran causes significantly higher concentrations . Diets containing these fibres (nominally 10% w:w) were administered for 3 weeks before, for 10 weeks during, and for 20 weeks after dimethylhydrazine administration, after which animals were killed and examined for tumours . Significantly fewer tumours were seen in the rats fed wheat bran compared with those fed guar or oat bran, and the total tumour mass was lowest in rats fed wheat bran . Rats on a 'no added fibre diet' had an intermediate tumour mass . Regression analysis, performed regardless of dietary group, showed that the concentration in stools of butyrate but not of acetate or stool volume, correlated significantly (and negatively) with tumour mass . These findings indicate that fibre which is associated with high butyrate concentrations in the distal large bowel is protective against large bowel cancer, while soluble fibres that do not raise distal butyrate concentrations, are not protective . Thus, butyrate production in vivo does bear a significant relationship to suppression of tumour formation. J Anim Sci, 1993 Mar, 71(3), 788 - 95 Periparturient changes in intake, ruminal capacity, and digestive characteristics in beef cows consuming alfalfa hay; Stanley TA et al.; Four multiparous, ruminally fistulated Angus x Hereford cows (average BW = 568 kg) were bred to the same bull and used to monitor periparturient changes in DMI, ruminal capacity, and digestion and fermentation characteristics . Cows were individually fed alfalfa hay (16.0% CP, 51.3% NDF) at 130% of the average DMI of the previous 5-d period . Ruminal capacity and DM fill were measured once every 2 wk by removing the ruminal contents from each cow and filling the empty rumen with water . Dry matter intake was measured daily . Ruminal VFA, pH, NH3, and total tract DM digestibility (DMD) were measured during three 7-d periods (average of 58 d before calving, 16 d before calving, and 25 d after calving) . Ruminal capacity and DM fill varied in a quadratic manner (P < .01); both were least 6 d before calving . Forage DMI (quadratic effect; P = .02) increased moderately during the prepartum period but increased dramatically after parturition . Although postpartum (d 22) ruminal capacity was only 5% greater than at 61 d before calving, postpartum DMI was 69% greater than DMI measured 61 d before calving . Indigestible ADF (IADF) passage rate changed quadratically with period (P = .01); the greatest IADF passage rate was observed 6 d before calving . Ruminal VFA (mM) also tended to change quadratically (P = .08); the highest concentrations occurred 25 d after calving . The acetate:propionate ratio declined linearly with period (P = .01) . Ruminal fluid dilution rate, pH, NH3, and DMD did not change significantly over time.(ABSTRACT TRUNCATED AT 250 WORDS) J Anim Sci, 1993 Mar, 71(3), 779 - 87 Influence of Aspergillus oryzae fermentation extract on forage intake, site of digestion, in situ degradability, and duodenal amino acid flow in steers grazing cool-season pasture; Caton JS et al.; Ten ruminally and duodenally cannulated (326 +/- 28 kg) and four esophageally fistulated (394 +/- 23 kg) steers grazing cool-season pasture throughout the growing season were used to evaluate the influence of Aspergillus oryzae fermentation extract (AO) supplementation on intake, forage nutrient utilization, and duodenal amino acid flow . Steers grazed a predominantly smooth brome (Bromus inermis L.) pasture, and measurements were taken in three periods (June, July, and August) . Steers were dosed daily at 0700 via the ruminal cannula with AO (2 g of AO per steer daily; DM basis) or not supplemented with AO . Each period consisted of 18 d for adaption to AO and 7 d for collection . Forage N was greater, and ADF was lower (P < .10), in June than in July and August . Ruminal pH, ammonia, total VFA concentration, and VFA proportions were not affected (P > .10) by AO supplementation . In vitro DM digestibility (percentage) and forage OM intake (grams/kilogram of BW) were greater (P < .10) for steers supplemented with AO . Ruminal and total tract NDF and ADF digestibilities were lower in June and greater during July (P < .10) in steers supplemented with AO . Total tract percentage of N disappearance was lesser during June and greater (P < .10) during July for steers receiving AO . Total, essential, and nonessential amino acid flows were increased (P < .10) by both AO supplementation and advancing season . In situ rate of ruminal CP degradation was not affected (P < .10) by treatment during any period.(ABSTRACT TRUNCATED AT 250 WORDS) Am J Gastroenterol, 1993 Mar, 88(3), 420 - 3 Fermentation of the carbohydrate of banana (Paradisiaca sapientum) in the human large intestine; Segal I et al.; Fermentation of dietary fiber and resistant starch is one of the major physiological functions of the human large intestine . The major substrate for fermentation is probably starch . This study assessed the effect of bananas--a carbohydrate with a highly resistant starch content--on breath hydrogen and methane production in methane and nonmethane-producing subjects . The results showed that both groups produced significant quantities of hydrogen after a banana meal, compared with a sucrose control test meal, measured as area under the curve (28 +/- 5.6 vs . 8.1 +/- 1.4 10(3) pm/min, p = 0.008 in methane producers and 39 +/- 15.2 vs . 10.5 +/- 4.1 10(3) ppm/min, p = 0.01 in methane nonproducers) . The rise in breath hydrogen started a half hour after the banana meal and peaked at 3 1/2 h in methane nonproducers, whereas in methane producers, the rise began after 2 h and peaked at 5 h . Methane production was not significantly stimulated by the test meals . This study shows that bananas stimulate fermentation mainly through the production of hydrogen, with minimal effect on methane production . The possible mechanisms for this process are discussed. Fetal Diagn Ther, 1993 Mar-Apr, 8(2), 74 - 8 Bacterial and fungal contamination of human fetal liver collected transvaginally for hematopoietic stem cell transplantation; Rice HE et al.; Transplantation of fetal hematopoietic stem cells is a new therapy for fetuses diagnosed in utero with inherited disorders . However, prior to transplantation of fetal stem cells, the cells must be free of microbial contamination . In order to investigate the contamination of human fetal liver tissue, we determined the rate and types of bacterial and fungal contamination of human fetal liver tissue collected transvaginally for use in stem cell transplantation by using the US Pharmacopoiea Assay . We found a high rate of contamination with bacteria or fungi (12 of the 14 fetal livers, or 85%) . Non-fermenting gram-negative rods were the predominant contaminants . The high rate of microbial contamination of fetal tissue suggests that techniques for tissue collection need to be improved before fetal stem cells are used for clinical transplantation. Appl Microbiol Biotechnol, 1993 Mar, 38(6), 770 - 5 High-level production and secretion of a mouse-human chimeric Fab fragment with specificity to human carcino embryonic antigen in Escherichia coli; Shibui T et al.; A high-level secretion system for the production of mouse-human chimeric antibody 21B2 (MHC 21B2) Fab fragment specific for human carcino embryonic antigen (hCEA) in Escherichia coli has been constructed . The genes encoding a light chain and an Fd fragment (a variable region and the CH1 domain of a heavy chain) of a mouse-human chimeric antibody were directly fused to the signal peptide of the E . coli ompF gene sequence . E . coli cells containing expression vectors in which each of the two genes are located downstream of a separate tac promoter were able to secrete the light chain and Fd fragment as two of their major cellular proteins . The signal peptides were efficiently removed from the primary products by post-translational processing, although they formed insoluble aggregates, possibly in the periplasm . In high-cell-density culture experiments using a jar fermentor, the amount of light chain and Fd fragment produced was at levels of up to 2.88 g/l and 1.28 g/l culture, respectively . By optimizing the conditions that encourage correct folding, formation of disulphide bonds, and association of the light chain with the Fd fragment, we have established a procedure that can purify, re-fold, and combine aggregated products to electrophoretically homogeneous Fab fragment with a yield of approximately 47% . Fab fragment produced in this manner shows essentially the same antigen-binding activity and specificity to hCEA as the parental mouse antibody 21B2 (MoAb 21B2). Appl Microbiol Biotechnol, 1993 Mar, 38(6), 719 - 27 High yield fermentation and purification of Tendamistat disulphide analogues secreted by Streptomyces lividans; Haas-Lauterbach S et al.; In our studies of structure-function correlation of polypeptides we used Tendamistat (TM), an alpha-amylase-inhibitor of Streptomyces tendae, as a model to investigate the influence of different mutants on the expression and secretion of the protein . In addition, we examined the influence of replacing the two disulphide-bridges that stabilize the two-loop structure of the whole protein . The single mutants C27S, C27T, C45A, the double mutants C11A/C27A, C11A/C27S, C11A/C27T, C11A/C27L, C45/C73A and the fourfold mutant C11A/C27A/C45A/C73A were prepared . The mutated TM gene was expressed in S . lividans TK 24, which secretes the active form of the inhibitor into the culture medium . Compared with the wild-type, the double-mutated TM derivatives show an increase in secreted protein by a factor of two to ten . In contrast, the single-mutated inhibitor analogues show the reverse effect . In order to examine the influence of temperature and culture media on the production of protein derivative we used the most unstable C11A analogue . Our expression studies at 10, 19, 28 and 37 degrees C established 19 degrees C as the optimal temperature for production of the protein derivatives . The correlation between the stability and secretion of TM is discussed in the context of our knowledge of protein translocation in bacteria . Based on these experiments we optimized the fermentation parameters, isolated TM analogous on a large scale, and verified them. Biotechnology (N Y), 1993 Mar, 11(3), 381 - 6 Construction of metabolic operons catalyzing the de novo biosynthesis of indigo in Escherichia coli; Murdock D et al.; The efficient production of the textile dye indigo by fermentation has been a goal since the early 1980's when the first bacterial strains capable of this synthesis were constructed . We report here the development of a recombinant microorganism that directly synthesizes indigo from glucose . This construction involved the cloning and genetic manipulation of at least 9 genes and modifications of the fermentation medium to help stabilize the biosynthetic activity . Directed genetic changes in two operons caused significant increases in reaction rates and in the stability of the catalytic enzymes . This example of whole cell catalysis by a recombinant Escherichia coli represents a novel and environmentally sound approach to the synthesis of a high value specialty chemical. Biotechnology (N Y), 1993 Mar, 11(3), 349 - 57 Process economics of animal cell and bacterial fermentations: a case study analysis of tissue plasminogen activator; Datar RV et al.; One link in the complex chain of medical economics is the cost of bringing new drugs and biologicals to the market . Advances in recombinant-DNA technology permit production of therapeutically active proteins in effectively unlimited quantities . Nevertheless, each expression system has a characteristic influence on the nature of the product produced and the process required to obtain it . In this case study we compare experiences with recombinant-tissue plasminogen activator (rtPA) produced in Chinese hamster ovary (CHO) cells and in Escherichia coli, with the aim of understanding the roles of some of the parameters that affect process economics . tPA belongs to the group of highly specific serine proteases that convert plasminogen to plasmin, which in turn degrades several protein substrates including fibrin, thus making it an effective thrombolytic agent . The treatment of acute myocardial infarction with such thrombolytic agents can result in early discharge of patients and decreased medical costs . However, there are major differences in the prices of the various available agents . The price of the FDA-licensed tPA product is $2,200 per dose or $22,000 per gram . It is believed that a significant portion of this price relates to manufacturing costs . We examine by way of case study illustration the cost breakdown for the two processes, and highlight important process, design and economic considerations that ultimately define a particular protein product. Mikrobiol Zh, 1993 Mar-Apr, 55(2), 99 - 104 {The inhibitory action of 6-azacytidine on Mollicutes and its proposed mechanism}; Skripal' IG et al.; 6-Azacytidine (6-AC) is shown to have an inhibitory effect on the Mollicutes of the different systematic position . The growth of type strains of Mollicutes (Acholeplasma laidlawii PG-8, Mycoplasma pneumoniae FH and M . fermentans PG-18) completely ceased in the nutrient medium at concentration of the above substance in it within the range of 125-250 micrograms/ml . 50% inhibiting concentration of 6-AC equaled: for M . fermentans PG-8: 23.43 micrograms/ml; M . pneumoniae FN: 46.8 micrograms/ml; Acholeplasma laidlawii PG-8: 62.5 micrograms/ml . 6-AC concentration 5 micrograms/ml decreased the process of DNA-dependent DNA synthesis in the in vitro system more than by 60% . 6-AC exerted less effect on the DNA-dependent RNA synthesis in the in vitro system: at different concentrations of 6-AC (up to 400 micrograms/ml) RNA synthesis decreased only by 20% . Translation on ribosomes of Mollicutes in the in vitro system completely ceased at 6-AC concentration 100 micrograms/ml . The results obtained indicate that for 6-AC in cells of Mollicutes and, possibly, for other microorganisms there are two targets: ribosomes and DNA-dependent DNA-polymerase . Total effect of blocking of the translation and replication processes by 6-azacytidine causes death of Mollicutes . Since 6-AC has no harmful effect on the human cells, it can be used as an efficient method for treatment of respiratory and urogenital diseases induced by Mollicutes. J Immunol Methods, 1993 Feb 26, 159(1-2), 229 - 34 Nanogram quantitation of secreted protein in a recombinant yeast fermentation using an immuno-ligand assay; Robinett RS et al.; A liquid-phase immuno-ligand assay has been developed for quantitative determination of recombinant tick anticoagulant protein (rTAP) secreted in yeast fermentations . A polyclonal anti-TAP antibody was labeled with biotin or fluorescein . Labelled antibodies were used in a non-competitive sandwich format to capture rTAP from solution, then reacted with urease-conjugated anti-fluorescein antibody . Detection of the immune complex was by a commercially available silicon-based potentiometric sensor which measures urease activity . Sample throughput was 90 samples per 7 h with a 2 h incubation time . The range of the standard curve was 0.1-10 ng/ml with an assay sensitivity of 0.025 ng/ml . For a mid-range concentration of 1 ng/ml, intraday and interday method precision was determined to be 1.031 +/- 0.061 and 1.077 +/- 0.026 ng/ml, respectively . Typically, spiked samples of 1 microgram rTAP/ml fermentation medium required dilutions of 1/1000 to generate a response in the mid-range of the standard curve . This assay provides a convenient method to quantitate product expression in multiple fermentation samples within 3 h after sampling . In addition, a modified version of the assay was developed which provided accurate results within 1 h of sample acquisition. Int J Exp Pathol, 1993 Feb, 74(1), 97 - 101 Factors influencing the ability of different mycoplasmas to colonize the genital tract of hormone-treated female mice; Furr PM et al.; Colonization of the genital tract of female mice, mainly BALB/c, by Mycoplasma genitalium, M . pneumoniae and M . pulmonis was enhanced by pretreatment of the mice with progesterone . M . fermentans, M . hominis and M . salivarium, and three serotypes and two untyped strains of Ureaplasma urealyticum colonized under the influence of oestradiol but not progesterone . Mycoplasmas dependent on progesterone were glucose-metabolizing, with strong haemadsorptive and other attachment properties, and possessed a terminal structure . Mycoplasmas dependent on oestradiol were arginine or arginine/glucose metabolizing . Ureaplasmas also required oestradiol . The oestradiol-requiring group appeared to be less cytadsorptive and devoid of a morphological terminal structure . Mycoplasmas that had had multiple passes in media were less able to colonize . This may be one, but not the only, reason for the failure of seven mycoplasmas to colonize under the influence of either hormone . The observations suggest the existence of a receptor mechanism for colonization, progesterone-requiring mycoplasmas being exposed to, and needing, genital tract cells different from those exposed to oestradiol-requiring mycoplasmas. J Antibiot (Tokyo), 1993 Feb, 46(2), 275 - 9 Three novel polyether antibiotics X-14889A, C, and D from a streptomycete . Taxonomy of the producing organism, fermentation production and biological properties of the antibiotics; Liu CM et al.; Antibiotic X-14889A, C, and D are novel polyether antibiotics related to lysocellin and antibiotic X-14873A . They are produced by a streptomycete isolated from a soil of Wisconsin . The antibiotic X-14889C is active against Gram-positive bacteria and exhibits ionophore property. J Antibiot (Tokyo), 1993 Feb, 46(2), 265 - 74 BMS-181184, a new pradimicin derivative . Screening, taxonomy, directed biosynthesis, isolation and characterization; Furumai T et al.; BMS-181184 is a new semisynthetic pradimicin derivative with a broad-spectrum antifungal activity . In a search for actinomycetes producing BMS-181184, 4 strains of Actinomadura sp . isolated from soil samples were found to produce the antibiotic under conditions of directed biosynthesis . Among them, Actinomadura sp . AB1236 proved most useful in the production of BMS-181184 when fermented in a medium containing D-serine and D-cycloserine . A minor product isolated from the broth of strain AB1236 was identified as the dexylosyl analog of BMS-181184, which was also obtained by acid hydrolysis of BMS-181184. J Antibiot (Tokyo), 1993 Feb, 46(2), 251 - 4 AB023, novel polyene antibiotics . I . Taxonomy of the producing organism, fermentation and antifungal activity; Cidaria D et al.; AB023 is a complex of polyene antibiotics produced by an actinomycete, SD581, which was isolated from a Kenyan soil sample . The two main components, pentaene antibiotics AB023a and AB023b, have antifungal activity against some phytopathogenic fungi, particularly against Botrytis cinerea (MIC of 5 micrograms/ml). J Antibiot (Tokyo), 1993 Feb, 46(2), 247 - 50 Mer-WF3010, a new member of the papulacandin family . I . Fermentation, isolation and characterization; Kaneto R et al.; Mer-WF3010, a new member of the papulacandin family, was isolated from the mycelia of Phialophora cyclaminis Mer-WF3010 (FERM P-11475) . The molecular formula of Mer-WF3010 was determined as C45H60O16. J Antibiot (Tokyo), 1993 Feb, 46(2), 222 - 8 Pepticinnamins, new farnesyl-protein transferase inhibitors produced by an actinomycete . I . Producing strain, fermentation, isolation and biological activity; Omura S et al.; Pepticinnamins A, B, C, D, E and F, a family of farnesyl-protein transferase (FPT) inhibitors were isolated from the fermentation broth of Streptomyces sp . OH-4652 . These inhibitors were purified from whole broth by extraction with chloroform, followed by silica gel column chromatography, Sephadex LH-20 chromatography and reverse phase HPLC . Among these, pepticinnamin C showed the most potent inhibition (IC50-100 nM). FEMS Microbiol Lett, 1993 Feb 1, 106(3), 233 - 7 Characterization of mutations that overcome the toxic effect of glucose on phosphoglucose isomerase less strains of Saccharomyces cerevisiae; Gamo FJ et al.; Glucose inhibits growth of yeast phosphoglucose isomerase mutants in permissive media . Mutants insensitive to this effect were isolated by selection on media containing 2% fructose + 2% glucose . A nuclear, monogenic, recessive mutation named rgl was responsible for this phenotype . The mutants isolated belonged to two complementation groups and have been termed rgl1 and rgl2 . When the double mutants were grown on fructose, fermentation of fructose or glucose was similar to that of the parental pgi strain but was not measurable when grown on fructose+glucose . Under these conditions, respiration of glucose and to a lesser extent of fructose was enhanced . The double mutants pgi rgl did not grow on fructose+glucose in the presence of antimycin A or ethidium bromide and their cytochrome oxidase was no longer sensitive to glucose repression . The results are interpreted as an indication that in the double mutants the glucose may be channeled through the pentose phosphate pathway to respiration. J Dairy Sci, 1993 Feb, 76(2), 514 - 24 Effects of supplemental protein source on intraruminal fermentation, protein degradation, and amino acid absorption; Keery CM et al.; Cannulated steers were used to determine the effects of supplemental soybean meal, heated soybean meal, fish meal, and a combination of fish meal, heated soybean meal, and corn gluten meal on intraruminal protein degradation and absorption of AA from the small intestine . Organic matter digestion in the reticulo-rumen was greater in steers fed diets supplemented with soybean meal, but whole tract digestibility was not affected by protein source . Total and bacterial CP flows to the abomasum were lower in steers fed diets supplemented with fish meal than in steers fed diets supplemented with heated soybean meal or the combination supplement . Dietary CP flow was 33.5% higher in steers fed diets supplemented with heated soybean meal than in steers fed diets supplemented with soybean meal, fish meal, or the combination supplement . Less essential and nonessential AA flowed to the abomasum and were absorbed from the small intestine of steers receiving diets supplemented with soybean meal . Digestibility of small intestine AA was 21.9% lower in steers receiving the soybean meal treatment . Abomasal flows of Met and Thr and absorption of Lys, Met, and Thr were increased in steers fed diets containing heated soybean meal, fish meal, and the combination supplement . These results suggest that the supply of AA deficient in microbial CP (Lys, Met, and Thr) can be increased and that absorbed AA balance can be changed markedly by selection of rumen escape protein supplements. An Esp Pediatr, 1993 Feb, 38(2), 107 - 12 {Relationship between primary lactose malabsorption and consumption of dairy products}; Escribano Subias J et al.; This study was designed to determine the influence of lactose malabsorption on the consumption of dairy products . We studied 157 children and 43 adults . The Breath-hydrogen test was used to define their level of lactose digestion . The prevalence of lactose maldigesters was 12% . We found a large relationship between the consumption of milk and milk products and age . Malabsorbers consumed more fermented dairy products (ripened cheese and yogurt) than did absorbers (p < 0.05) . Subjects with normal lactose absorption consumed more milk, butter, cream cheese and global lactose than the maldigesters (p < 0.05) . Lactose intolerance, familiar consumption and geographic origins had little influence on an individual's consumption habits. Appl Environ Microbiol, 1993 Feb, 59(2), 405 - 9 Inhibition of ruminal cellulose fermentation by extracts of the perennial legume cicer milkvetch (Astragalus cicer); Weimer PJ et al.; Cicer milkvetch (Astragalus cicer L.) is a perennial legume used as a pasture or rangeland plant for ruminants . A study was undertaken to determine whether reported variations in its ruminal digestibility may be related to the presence of an antinutritive material . In vitro fermentation of neutral detergent fiber (NDF) of cicer milkvetch by mixed rumen microflora was poorer than was the fermentation of NDF in alfalfa (Medicago sativa L.) . Fermentation of cicer milkvetch NDF was improved by preextraction of the ground herbage with water for 3 h at 39 degrees C . Such water extracts selectively inhibited in vitro fermentation of pure cellulose by mixed ruminal microflora and by pure cultures of the ruminal bacteria Ruminococcus flavefaciens FD-1 and Fibrobacter succinogenes S85 . Inhibition of the cellulose fermentation by mixed ruminal microflora was dependent upon the concentration of cicer milkvetch extract and was overcome upon prolonged incubation . Pure cultures exposed to the extract did not recover from inhibition, even after long incubation times, unless the inhibitory agent was removed (viz., by dilution of inhibited cultures into fresh medium) . The extract did not affect the fermentation of cellobiose by R . flavefaciens but did cause some inhibition of cellobiose fermentation by F . succinogenes . Moreover, the extracts did not inhibit hydrolysis of crystalline cellulose, carboxymethyl cellulose, or p-nitrophenylcellobioside by supernatants of these pure cultures of cellulolytic bacteria or by a commercial cellulase preparation from the fungus Trichoderma reesei . The agent caused cellulose-adherent cells to detach from cellulose fibers, suggesting that the agent may act, at least in part, by disrupting the glycocalyx necessary for adherence to, and rapid digestion of, cellulose. Biotechniques, 1993 Feb, 14(2), 228 - 33 An autosampler for fermentation; Page WJ et al.; The operation of an autosampler, constructed from a peristatic pump, an interval timer and a fraction collector, for the removal of whole broth fermentation samples is described . The autosampler repetitively removes samples of user-determined size at repetitive time intervals . The sampler is intended for use with low containment fermentations . A comparison of the autosampler and the manual sampler of a fermenter showed good correlation of substrate consumption rates when changes were sizeable and superior results were obtained with the autosampler when the changes were subtle during a typical diauxic fermentation of Azotobacter vinelandii. J Nutr, 1993 Feb, 123(2 Suppl), 418 - 23 Colon cancer--do the nutritional epidemiology, the gut physiology and the molecular biology tell the same story? Potter JD. Colon carcinogenesis models exist at epidemiologic, physiologic and molecular biologic levels . Thinking about the coherence of such models is useful both to inform colon cancer research and because the reasoning process may be generalizable . The consistent epidemiologic risk factors are low vegetable/fiber and high fat/meat/protein intakes . Others include physical activity, alcohol and reproduction . These epidemiologic risk factors appear to map to physiologic variables that provide mechanistic explanations for the associations: higher bile acids, fiber fermentation and effects of specific anticarcinogens found in vegetables . The possibility that the meat/fat association is due to carcinogens or promoters produced in cooked foods adds complexity to the physiologic model . As a link across genetics, physiology and epidemiology, the role of acetylator status is considered . Finally, whether relationships might exist between the epidemiologic/physiologic risk factors and the recently described molecular genetic changes and other colon cancer molecular mechanisms is considered. Protein Expr Purif, 1993 Feb, 4(1), 52 - 8 A purification method for labile variants of ribonuclease T1; Mayr LM et al.; We present a new procedure for the rapid production of ribonuclease T1 variants with decreased stability which could not be purified in satisfying amounts by the existing methods . The major changes from the established procedures are the following . (i) The cells were grown at 28 degrees C rather than at 37 degrees C . (ii) The entire purification was performed at low temperatures (4 degrees C) . (iii) Materials for chromatography with high flow rates were used to accelerate protein isolation . (iv) The pH was lowered from 7.5 to 6.0, a condition under which RNase T1 is much more stable . The use of this improved procedure allowed the purification of the labile P39G and P73V variants of RNase T1 . By the same technique 300 mg of the wild-type protein could be isolated from 10 liters liquid culture within 3 days . The P39G and the P73V mutations strongly decrease the stability of RNase T1 and the midpoints of the reversible thermal unfolding transition are lowered by 16 and 6 degrees C, respectively, relative to that of the wild-type protein . The decrease in temperature during fermentation and the rapid purification at low temperature and under solvent conditions where the stability of the proteins is high are probably the major reasons for the dramatic increase in yield of these labile variants of RNase T1 . Such an approach should be valuable for the production of recombinant proteins in general. J Bacteriol, 1993 Feb, 175(3), 870 - 8 Anaerobic regulation of the adhE gene, encoding the fermentative alcohol dehydrogenase of Escherichia coli; Leonardo MR et al.; The regulation of the adhE gene, which encodes the trifunctional fermentative acetaldehyde-alcohol dehydrogenase of Escherichia coli, was investigated by the construction of gene fusions and by two-dimensional protein gel electrophoresis . Both operon and protein fusions of adhE to lacZ were induced 10- to 20-fold by anaerobic conditions, and both fusions were repressed by nitrate, demonstrating that regulation is at the level of transcription . Nitrate repression of phi (adhE-lacZ) expression, as well as of alcohol dehydrogenase enzyme activity, was partly relieved by a mutation in narL . Mutations in rpoN or fnr had no effect on the expression of adhE . Two-dimensional protein gels demonstrated that increases in the amount of adhE protein correlated with increases in enzyme activity, demonstrating that induction was due to synthesis of new protein, not to activation of preexisting protein . When oxidized sugar derivatives such as gluconate or glucuronate were used as carbon sources, the anaerobic expression of phi (adhE-lacZ) was greatly reduced, whereas when sugar alcohols such as sorbitol were used, the expression was increased compared with expression when glucose was the carbon source . This observation suggested that induction of phi (adhE-lacZ) might depend on the level of reduced NADH, which should be highest with sorbitol-grown cells and lowest with glucuronate-grown cells . When phi (adhE-lacZ) was present in a strain deleted for the adhE structural gene, anaerobic expression of phi (adhE-lacZ) was approximately 10-fold higher than in an adhE+ strain . Since the presence of alcohol dehydrogenase would serve to decrease NADH levels, this finding again implies that the adhE gene is regulated by the concentration of reduced NAD . Introduction of a pgi (phosphoglucose isomerase) mutation reduced the anaerobic induction of phi(adhE-lacZ) when the cells were grown on glucose, but had little effect on fructose-grown cells . Pyruvate did not overcome the pgi effect, but glycerol 3-phosphate did, which is again consistent with the possibility that adhE expression responds to the level of reduced NAD rather than to a glycolytic intermediate. J Antibiot (Tokyo), 1993 Feb, 46(2), 207 - 13 SCH 45752--an inhibitor of calmodulin-sensitive cyclic nucleotide phosphodiesterase activity; Hegde VR et al.; A highly potent inhibitor of calmodulin-sensitive phosphodiesterase (PDE) activity was isolated from the culture broth of an unidentified fungal isolate, SCF-125 . A chemically defined medium was developed for production of this compound . The PDE inhibitor was isolated from the fermentation filtrate by adsorption on a macro-reticular resin and further purified by gel filtration chromatography and reverse-phase HPLC . The major PDE inhibitor was identified as cephalochromin, a bis-naphthopyrone, by spectral data analysis . The compound, SCH 45752, inhibited calmodulin-sensitive PDE activities with IC50 values of 40-47 nM . It inhibited the activities of calmodulin-independent PDE and various protein kinases with higher IC50 values (2-40 microM) . SCH 45752 does not appear to be a calmodulin antagonist . Furthermore, SCH 45752 affects smooth muscle contraction at a concentration of 30 microM; it potentiated the relaxing effect of sodium nitroprusside on carotid artery media contracted by histamine . Thus SCH 45752 is one of the most potent inhibitors of calmodulin-sensitive PDE activity known, and it is capable of exerting a pharmacological effect in at least one intact tissue model. Eur J Biochem, 1993 Feb 1, 211(3), 697 - 702 Cloning, sequencing and expression of the gene encoding the carboxytransferase subunit of the biotin-dependent Na+ pump glutaconyl-CoA decarboxylase from Acidaminococcus fermentans in Escherichia coli; Bendrat K et al.; 1 . The primary sodium-ion pump glutaconyl-CoA decarboxylase (GCD) from Acidaminococcus fermentans is composed of four subunits: GCDA, the carboxytransferase (65 kDa), GCDB, the carboxylyase (36 kDa), GCDC, the biotin carrier (24 kDa) and GCDD (14 kDa) of unknown function . A genomic library of A . fermentans was screened with an antiserum raised against whole GCD . A clone giving the strongest reaction in an immunoassay contained a 12-kbp genomic fragment from A . fermentans and was analysed further . An oligonucleotide deduced from the N-terminus of GCDA was used for probing the corresponding gene gcdA . It is 1761 bp in length and encodes for a protein of 64.3 kDa . Both partial amino acid sequences obtained from GCDA, the N-terminus as well as an internal tryptic peptide, were detected in the open reading frame (ORF) of gcdA . 2 . Sequencing of the flanking regions revealed three adjacent ORF (ORF1-3) which do not code for any of the peptide sequences known of the other GCD subunits . The ORF downstream of gcdA (ORF3) is followed by hgdA and hgdB coding for 2-hydroxyglutaryl-CoA dehydratase, the preceding enzyme of the pathway of glutamate fermentation . Our results suggest that at least these three genes of the hydroxyglutarate pathway are organised in an operon and that the genes of the other GCD subunits from which peptide sequences are known (GCDB and GCDC) are not located adjacent to gcdA . 3 . gcdA was amplified from genomic DNA using the polymerase chain reaction and cloned into the expression vector pJF118HE . Active GCDA subunit (up to 2.8 nkat/mg protein), catalysing the biotin-dependent formation of crotonyl-CoA from glutaconyl-CoA, was obtained in cell-free extracts of Escherichia coli DH5 alpha by moderately inducing the tac promoter of pJF118HE with 25-100 microM isopropyl-1-thio-beta-D-galactoside . Strong induction (1 mM isopropyl-1-thio-beta-D-galactoside) led to the formation of inclusion bodies from which GCDA could not be reactivated . The apparent Km = 51 mM for free biotin of the expressed GCDA subunit with V = 1.9 nkat/mg protein is similar to that of butanol-treated GCD composed of GCDA and GCDC (apparent Km = 40 mM) . Biocytin was found to be a somewhat better carboxy acceptor for the expressed GCDA subunit (apparent Km = 13 mM; V = 1.0 nkat/mg protein) . 4 . Native GCD and expressed GCDA were treated with 2 mM N-ethylmaleimide showing different kinetics of inactivation: GCD lost half of its activity within 6 min, whereas expressed GCDA required 21 min. J Nutr, 1993 Feb, 123(2), 244 - 52 Viscosity and fermentability as attributes of dietary fiber responsible for the hypocholesterolemic effect in hamsters; Gallaher DD et al.; The attribute(s) of soluble dietary fibers responsible for cholesterol lowering is currently uncertain . A series of experiments were conducted in which viscosity and fermentability was assessed independently for their effect on plasma and liver cholesterol concentration . Hamsters were divided into four dietary groups and fed diets containing 0.12% cholesterol and 5% fiber as high viscosity hydroxypropyl methylcellulose (HV-HPMC group), low viscosity hydroxypropyl methylcellulose (LV-HPMC group), high viscosity guar gum (HV-GG group) or low viscosity guar gum (LV-GG group) . Hydroxypropyl methylcellulose is essentially nonfermentable, whereas guar gum is highly fermentable . Plasma cholesterol concentrations at 3, 6 and 11 wk and liver cholesterol concentrations at 6 and 11 wk were significantly lower in the HV-HPMC group relative to the LV-HPMC group (P < 0.05) . Intestinal content viscosities of the LV-HPMC and HV-GG groups were similar; consequently, these two groups were compared to examine the independent effect of fermentation . Plasma and liver cholesterol were significantly lower in the HV-GG group compared with the LV-HPMC group at 6 wk (P < 0.05), but not at 3 or 11 wk . Hepatic sterol synthesis rates were not affected by any of the diets . This study shows that greater viscosity of intestinal contents is strongly associated with cholesterol reduction, but that the contribution of fiber fermentation remains uncertain. Microbiologia, 1993 Feb, 9 Spec No, 76 - 82 {Utilization of molecular techniques for the characterization of wine yeasts and the study of the wine-making process}; Querol A et al.; The study of the fermentation process in the Alicante region allowed us to conclude that adverse climatic conditions could be responsible for deficient wine-making with serious problems arising fermentations, usually causing incomplete fermentation . In order to avoid these problems, we selected a Saccharomyces cerevisiae strain, namely T73, isolated in the same Alicante region, to be used to perform controlled fermentations . The use of selected strains in the wine-making process requires the development of characterization techniques that can clearly differentiate between the inoculated strain and the wild strains present in the musts . In order to differentiate strains present in the wine ecosystems, an extensive survey of different methods of yeast strains identification has been carried out . However, these techniques are very complex to be used in industry . For this reason, we have developed a new, simple, inexpensive and rapid method based on mitochondrial DNA restriction analysis . This technique was applied to the control of wine fermentations conducted by active dry yeasts . This molecular approach allows us to understand the role of the inoculated dry yeast strain and that of the natural S . cerevisiae flora during wine fermentation. Comp Biochem Physiol Comp Physiol, 1993 Feb, 104(2), 357 - 60 Changes in food intake with ambient temperature alter hindgut fermentation in the damara mole-rat Cryptomys damarensis; Yahav S et al.; 1 . Changes in food consumption and fermentation capacity were examined in mole-rats housed at 23 and 30 degrees C . 2 . At both temperatures animals maintained body mass . Food consumption was lower at the higher temperature and led to a similar decline in caecal mass . 3 . Caecal dry matter content and fermentation efficiency were unaffected by food intake . However, gas production per animal decreased markedly with the reduced food intake . 4 . The drop in fermentation in response to decreased food intake led to a concomitant decrease in heat production . This would be highly advantageous for chthonic rodents residing during summer in warm humid plugged burrows. J Ind Microbiol, 1993 Feb, 12(2), 99 - 102 The effect of cerulenin on the production of esperamicin A1 by Actinomadura verrucosospora; Lam KS et al.; Addition of cerulenin (0.25-1.0 mM) to cultures of Acinomadura verrucosospora before the onset of esperamicin synthesis inhibited the production of esperamicin A1 by the microorganism . This result indicates that esperamicin A1 is biosynthesized in part by the polyketide pathway . Addition of cerulenin to the cultures during the active production phase led to a net decrease in esperamicin A1 production . The 14C-acetate labeling pattern of esperamicin A1 in the cultures with or without addition of cerulenin at the active production phase also demonstrated the instability of esperamicin A1 in the fermentation . This suggests that esperamicin A1 is unstable and degradation occurs during the active production phase . Addition of the neutral resin Diaion HP-20 (1%) to the fermentation enhanced the production of esperamicin A1 by 53%. J Ind Microbiol, 1993 Feb, 11(2), 95 - 103 Improvement in the titer of echinocandin-type antibiotics: a magnesium-limited medium supporting the biphasic production of pneumocandins A0 and B0; Tkacz JS et al.; We have developed a liquid fermentation medium for the submerged culture of the fungus, Zalerion arboricola, which supports the rapid production of an echinocandin-type antibiotic, pneumocandin A0 (formerly L-671,329), in yields increased at least 4-fold over those reported previously . The improvements were achieved through medium simplification, substitution of high levels of mannitol for glycerol as the major source of carbon, and restriction of available magnesium . Antibiotic formation in batch cultures with this mannitol-based medium is not confined to the idiophase; rather production appears to be biphasic, with synthesis beginning during growth (i.e., at day 3) and increasing in rate at day 11, well after rapid growth has ended . Accumulation of antibiotic continues beyond 14 days, and by 21 days titers greater than 500 micrograms/ml are attained . For the synthesis of a related compound, pneumocandin B0, by a mutant strain of Z . arboricola, the medium gives similar production kinetics and a titer of 800 micrograms/ml . Although supplementation of the medium with magnesium ions stimulates growth, it decreases titer by preferentially affecting the second phase of antibiotic synthesis . This decline in synthesis in the magnesium-supplemented medium is explained by the depletion of mannitol before the second phase of synthesis can begin . In contrast, mannitol in the magnesium-limited medium is used more slowly with approximately half still available at day 11 to support continued antibiotic formation. Biotechnology (N Y), 1993 Feb, 11(2), 207 - 12 A novel process for the large-scale purification of recombinant tick anticoagulant peptide using perfusion chromatography; Lehman ED et al.; Tick anticoagulant peptide (TAP) is a 60 amino acid peptide (Mr = 6977, pI = 4.9) found in the saliva of the soft tick Ornithodorous moubata that specifically inhibits blood coagulation factor Xa (fXa) . A recombinant form of TAP (rTAP) secreted by Saccharomyces cerevisiae was purified from 200 liters of fermentation broth with POROS, strong cation exchange (SCX) and reversed-phase high performance liquid perfusion chromatography (RP-HPLC) media (20 microns nominal particle diameter) . The higher linear flow rates and dynamic capacities, as well as low back pressures, obtained with perfusion chromatography media permitted 37.5 g of rTAP to be efficiently captured and significantly enriched from 400 liters of diafiltered fermentation broth (24.3 g yield) with 1.25 liter of SCX media using a low pressure column and peristaltic pump in 4.5 hours . Subsequently, 16.7 g of rTAP obtained from the capture step was purified in a high-resolution mode with the same SCX media, after the media was cleaned and repacked into an 800 ml high-pressure column . By doing multiple rapid cycles at high linear flow rates, preparative-scale high-resolution purification of multi-gram amounts of the peptide was done on this relatively small column in only 10.5 hours . Finally, the peptide was desalted and decolorized on a 200 ml RP-HPLC column of perfusion chromatography media by doing multiple rapid cycles . After lyophilization, 12 g of peptide (46.9% yield) was obtained that was > 96% homogeneous by several analytical criteria and fully active in inhibiting blood coagulation factor Xa (fXa).(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1993 Jan 25, 268(3), 1824 - 9 Cloning and sequencing of a cDNA encoding Saccharomyces cerevisiae carnitine acetyltransferase . Use of the cDNA in gene disruption studies; Kispal G et al.; cDNA encoding for carnitine acetyltransferase (CAT) of yeast S . cerevisiae was isolated by screening a yeast cDNA lambda gt11 library with antibody . The whole coding sequence was obtained from the cDNA and from a YEP 13 DNA clone identified using the cDNA as probe . The coding sequence consists of 670 residues, which amounts to a molecular mass of 77,300 kDa . This cDNA was used successfully to disrupt the gene for the mitochondrial isoenzyme of CAT, which was shown by measuring the enzyme activity and by immunoblot . The acetylcarnitine content of these cells decreased significantly . A search in the PIR protein data base revealed that besides the known carnitine acyltransferases, choline acyltransferases are highly homologous to yeast CAT . The mitochondrial CAT-deficient (CAT-) cells were able to grow on different fermentable and nonfermentable carbon sources, even on acetate at the same rate as the parental strain . In contrast to these, 13C NMR studies revealed significant differences between parental and CAT- cells . In CAT-cells {3-13C}pyruvate was converted mainly to lactate and acetate, whereas in the parental cells alanine and tricarboxylic acid cycle intermediates were found as the main products of pyruvate metabolism beside acetate . These results suggest diminished flux through the pyruvate dehydrogenase complex in the absence of mitochondrial CAT in yeast cells. FEMS Microbiol Lett, 1993 Jan 15, 106(2), 201 - 4 Mycoplasma cells stimulate in vitro activation of plasminogen by purified tissue-type plasminogen activator; Tarshis M et al.; In an in vitro direct assay with tissue-type plasminogen activator (tPA), plasminogen and the chromogenic substrate S-2251, the ability of Mycoplasma fermentans KL4 to stimulate tPA-mediated activation of plasminogen to plasmin was studied . Mycoplasma cells markedly enhanced the activation of plasminogen by tPA in a concentration-, temperature- and pH-dependent manner . Nonidet P-40 (0.01%), sonication, and freezing and thawing of the cells substantially increased the stimulatory effect of mycoplasma on tPA activity . In contrast, the activation of plasminogen by urokinase was refractory to mycoplasma cells . The mycoplasma-mediated stimulation of tPA activity was prevented by epsilon-aminocaproic acid (EACA), a lysine analogue known to block lysine-binding sites (LBS) in plasminogen and tPA . Among several Mycoplasma fermentans strains tested, incognitus strain demonstrated the highest stimulation activity . These results suggest that mycoplasma cells interact with LBS in tPA and plasminogen to enhance plasminogen activation. Biochim Biophys Acta, 1993 Jan 15, 1161(1), 19 - 27 Purification and characterization of pyruvate ferredoxin oxidoreductase from the hyperthermophilic archaeon Pyrococcus furiosus; Blamey JM et al.; Pyrococcus furiosus grows optimally at 100 degrees C by carbohydrate fermentation . It is thought to contain a novel tungsten-dependent, NAD(P)-independent glycolytic pathway in which one of the oxidation steps is catalyzed by a tungsten-containing aldehyde ferredoxin oxidoreductase . The enzyme that catalyzes the terminal oxidation step, pyruvate ferredoxin oxidoreductase (POR), has now been purified . POR has a molecular mass of 100 kDa and is comprised of three subunits (45, 31 and 24 kDa) . It lacks tungsten but contains thiamine pyrophosphate (TPP) and two ferredoxin-type {4Fe-4S} clusters per molecule which, by EPR spectroscopy, can be differentiated by their relaxation properties . The enzyme requires CoASH but not TPP for pyruvate oxidation activity and will not use 2-oxoglutarate, phenyl pyruvate or indole pyruvate as substrates . POR is virtually inactive at 25 degrees C and shows a temperature optimum for pyruvate oxidation above 90 degrees C . The apparent Km values for pyruvate, CoASH and P . furiosus ferredoxin at 80 degrees C are 460, 100 and 70 microM, respectively . Carbon monoxide was a potent inhibitor of pyruvate oxidation (apparent Ki = 7 microM) . The half-life of activity (t50%) in air at 25 degrees C was 15 min and the t50% value at 80 degrees C (under anaerobic conditions) was 23 min . Based on molecular comparisons with PORs from mesophilic organisms, it is proposed that P . furiosus POR may represent an ancestral form of a pyruvate-oxidizing enzyme. Ugeskr Laeger, 1993 Jan 4, 155(1), 30 - 2 {A stink bomb in an office environment . Sick building syndrome with toxic rhinitis after exposure to fusel}; Hein HO et al.; In 1983 WHO, defined "The Sick Building Syndrome" . Various conditions influence the indoor climate, among others the degassing of chemicals . An epidemic of sick building syndrome is described in a two year old office building . The symptoms appeared after exposure to toxic chemicals released by a stink bomb--a form of exposure not previously described in the literature . Gas chromatographic analysis of the content of the stink bomb revealed 22 different chemicals likely to be remains from an alcoholic fermentation process . Twenty-four employees were exposed . A questionnaire investigation of the employees revealed that seven had symptoms related to the exposure . A clinical investigation of those who claimed to have symptoms took place . Six of the seven patients were investigated . They all had toxic rhinitis with bleeding . Owing to an unsystematic procedure it took more than two months before the indoor climate was normalized . The toxic rhinitis and other symptoms gradually decreased over more than four months . In order to minimize potential health damage due to the sick building syndrome, we recommend that experts should be consulted within this particular field. Arch Tierernahr, 1993, 43(1), 79 - 90 {Ruminal nitrogen metabolism in calves and sheep . 3 . Effect of hay-concentrate ratio in calves}; Boldt E et al.; During the milk feeding period (ca . 55th day of life) and 2 weeks after the milk feeding period (ca . 80th day of life) the influence of a hay quota in the ration (on DM basis) of 6, 10, 14 and 18% in the milk feeding period and of 16, 23, 31 and 37% in the period after milk feeding was tested as to ruminal N metabolization with calves supplied with duodenal re-entrant cannulae . The increase of the hay quota in the ration for ruminating calves reduced as a tendency duodenal NAN passage, its bacterial N-quota, post-ruminal NAN digestibility and, due to a worse utilization of available N for bacterial N-synthesis, bacterial N-synthesis rate . The apparent fermentation of organic matter was, as a tendency, increased with the growing hay quota in the ration. Vet Med (Praha), 1993, 38(4), 229 - 35 {The effect of cadmium on indicators of rumen fermentation and their levels in biological material in sheep}; Sviatko P et al.; The experiment was carried out with three groups, each consisting of six adult sheep of the Merino breed weighing 40-45 kg, fed with the diet composed of 1 kg of meadow hay and an addition of 0.2 kg of barley meal for 3 weeks before the experiment . The first group - the control - was provided the natural supply 0.09 mg Cd/kg DM of the diet, the supply of Ist experimental group was increased by 5 mg Cd/kg DM and of the IInd experimental group by 10 mg Cd/kg of diet DM in the form of sulphate . The influence of mentioned cadmium doses on levels of volatile fatty acids, their energetic yield and proportion of their energy and energy of methane and bacterial mass from the energy of fermented feed hexose, as well as the total number of infusoria and number of infusoria of Entodinium sp . in the ruminal fluid of sheep with an increased supply of cadmium were examined during seven weeks . In addition to that cadmium contents in the liver, kidneys, muscles, excrements and ruminal fluid was investigated . Obtained results indicate that yet a lower supply of cadmium amounting to 5 mg and 10 mg/kg DM of the sheep diet, had significant negative influence (Tab . I) on the production of isobutiric acid in rumen fluids and decreased significantly (Tab . III) the total counts of infusoria and number of Entodinium sp . infusoria in the rumen fluids of sheep.(ABSTRACT TRUNCATED AT 250 WORDS) Nahrung, 1993, 37(2), 141 - 6 Effect of rabadi fermentation on phytic acid and in vitro digestibility of barley; Gupta M et al.; Rabadi, an indigenous fermented food, was prepared by mixing cereal flour with buttermilk, allowing it to ferment at 30, 35 and 40 degrees C for 6, 12, 18, 24 and 48 h and cooking the fermented mixture for 0.5 h with continuous stirring . Two types of rabadi were prepared i.e . autoclaved and unautoclaved . In autoclaved type of rabadi cereal flour was mixed with water, autoclaved (0.103 MPa = 15 psi for 15 min), cooled, mixed with buttermilk and fermented . As this type of rabadi was precooked prior to fermentation, hence, the fermented product did not require cooking afterwards, while in unautoclaved rabadi, barley flour and buttermilk were mixed, fermented and then cooked prior to consumption . Phytic acid was reduced drastically at all the temperatures and periods of fermentation in both autoclaved and unautoclaved type of rabadi; greater reduction occurred at higher temperature and duration of fermentation . A significant improvement in the in vitro digestibility of starch and protein was observed; maximum improvement was noticed when fermentation was carried out at 40 degrees C for 48 h in both the types of rabadi . Phytic acid had a significant (P < 0.05) negative correlation with digestibility (in vitro) of proteins and starch of barley flour rabadi. Schweiz Arch Tierheilkd, 1993, 135(4), 117 - 24 {The "hemorrhagic bowel syndrome" of swine: clinical, pathologo-anatomic and etiopathogenic aspects}; Hani H et al.; In an analysis of autopsy findings from 16,384 pigs (1980-90) 436 cases of haemorrhagic bowel syndrome (HBS) were found (2.66% of autopsies) . In most cases fattening pigs (in the weight of 25-100 kg) were affected . HBS was significantly more frequent in females . Intestinal volvulus was confirmed in 56.2% of all cases of HBS, however by more careful examination between 1988 and 1990 even in 80% of cases . In most cases the degree of torsion was 180 degrees and the direction as seen with the pig lying on its back was anticlockwise . Clinical history as reported by owners revealed limited information: sudden death of one or several pigs within three months, association with whey feeding not uncommon . Significantly more cases of HBS were seen during spring and on mondays . Numerous yeasts could be detected in mucosal impression smears of ileum, colon and caecum . Additional analyses in six farms suffering from big losses due to HBS suggested that several environmental and management factors may be involved pathogenetically: feeding only once a day of excessive amounts of a liquid diet, especially whey (a highly fermentable substrate), poor hygiene of liquid diets (high bacterial counts and yeast concentrations) . In mixed breeding and fattening units, the aforementioned factors could be responsible also for sudden deaths in dry sows caused by colonic bloat often associated with intestinal rupture and torsions of the stomach and/or the spleen. Vopr Med Khim, 1993 Jan-Feb, 39(1), 45 - 7 {Comparative physico-chemical study of L-lysine-alpha-oxidase from surface and fermenter growth Trichoderma sp.}; Lukasheva EV et al.; Homogeneous preparations of L-lysine-alpha-oxidase were obtained from Trichoderma sp cultivated by using a surface technique and a fermenter set . The homogeneous enzyme preparations were similar in molecular mass, isoelectric point and substrate specificity . There was the single difference in the absorbance spectra, which may occur due to the presence of cofactor FAD in various oxidation states . The findings suggest that cultivation of Trichoderma sp in the fermenter set did not alter properties of L-lysine-alpha-oxidase produced. Curr Genet, 1993, 23(4), 281 - 9 The growth and signalling defects of the ggs1 (fdp1/byp1) deletion mutant on glucose are suppressed by a deletion of the gene encoding hexokinase PII; Hohmann S et al.; Yeast cells defective in the GGS1 (FDP1/BYP1) gene are unable to adapt to fermentative metabolism . When glucose is added to derepressed ggs1 cells, growth is arrested due to an overloading of glycolysis with sugar phosphates which eventually leads to a depletion of phosphate in the cytosol . Ggs1 mutants lack all glucose-induced regulatory effects investigated so far . We reduced hexokinase activity in ggs1 strains by deleting the gene HXK2 encoding hexokinase PII . The double mutant ggs1 delta, hxk2 delta grew on glucose . This is in agreement with the idea that an inability of the ggs1 mutants to regulate the initiation of glycolysis causes the growth deficiency . However, the ggs1 delta, hxk2 delta double mutant still displayed a high level of glucose-6-phosphate as well as the rapid appearance of free intracellular glucose . This is consistent with our previous model suggesting an involvement of GGS1 in transport-associated sugar phosphorylation . Glucose induction of pyruvate decarboxylase, glucose-induced cAMP-signalling, glucose-induced inactivation of fructose-1,6-bisphosphatase, and glucose-induced activation of the potassium transport system, all deficient in ggs1 mutants, were restored by the deletion of HXK2 . However, both the ggs1 delta and the ggs1 delta, hk2 delta mutant lack detectable trehalose and trehalose-6-phosphate synthase activity . Trehalose is undetectable even in ggs1 delta strains with strongly reduced activity of protein kinase A which normally causes a very high trehalose content . These data fit with the recent cloning of GGS1 as a subunit of the trehalose-6-phosphate synthase/phosphatase complex.(ABSTRACT TRUNCATED AT 250 WORDS) Reprod Nutr Dev, 1993, 33(1), 43 - 9 Effect of a viable yeast culture on digestibility and rumen fermentation in sheep fed different types of diets; Fiems LO et al.; Five mature wethers fitted with rumen fistulas were fed grass hay and a sugarbeet-pulp-based concentrate or maize silage and a cereal-based concentrate (50/50 digestible organic matter basis), or without with 5 g yeast supplement (Saccharomyces cerevisiae, Biosaf) per day in a latin square design . Diets were given for a 28-d adaptation period, followed by a 10-d collection period to determine digestibility and nitrogen retention data . Afterwards, rumen samples were taken on 3 consecutive days and analysed for volatile fatty acids, pH and ammonia . Digestibility and nitrogen balance were not affected by yeast treatment . Supplementation of yeast increased acetate: propionate ratio, butyrate, isoacids, pH and ammonia . The effects were more pronounced for the maize silage diet . These results demonstrate that the effect of yeast culture on rumen fermentation may depend on the nature of the diet . Living yeast cell number in the rumen fluid rapidly declined when dietary yeast was ceased . Furthermore, yeast cells survived the passage through the digestive tract. Mikrobiol Zh, 1993 Jan-Feb, 55(1), 12 - 8 {The isolation and purification of a nonspecific DNAse from Mycoplasma fermentans PG-18}; Sitailo SZ et al.; Nonspecific endogenic DNAase has been isolated from biomass of Mycoplasma fermentans PG-18 cells and purified to the homogeneous state . The scheme of isolation consists of purification stages on columns with phosphocellulose, DNA-cepharose CL-8B and phenylcepharose . DNAase was not bound to phosphocellulose, its volume was equal to zero . Then this DNAase was passed through column with DEA-cepharose CL-6B (elution by gradient KCl from 0.1 to 1.8M): enzyme was eluted at KCl concentration in the eluting buffer from 0.1 to 1.2 M . The enzyme was purified to the homogeneous state on column with phenylcepharose (elution by linear gradient of ethylene glycol from 30 to 80%): enzyme was eluted at the concentration of ethylene glycol in the eluting buffer from 43 to 80% . According to data obtained using gel-electrophoresis, under the denaturing conditions molecular mass of enzyme in acrylamide gel was 34 kDa. Yeast, 1993 Jan, 9(1), 85 - 94 Yeast flocculation: flocculation onset and receptor availability; Stratford M; Flocculent strains of brewing yeast grow and ferment as single cells and flocculate in the stationary phase of growth . The switch from single-celled yeast growth to multi-celled aggregation, flocculation onset, is of critical importance to the brewing industry . Yeast flocculation involves adhesion of surface-lectins on flocculent cells to carbohydrate receptors on neighbouring cell-walls . The presence of carbohydrate receptors, outer-chain mannan side-branches, was monitored throughout growth of flocculent and non-flocculent strains of Saccharomyces cerevisiae, with particular attention to the growth phases where flocculation is normally developed . Receptors were probed by coflocculation with flocculent strains and by aggregation with concanavalin A, a lectin shown to use the same receptors as flocculation . While considerable variation was found in coflocculation and concanavalin A aggregation between strains, little or no change in receptor availability was found throughout the growth of all yeast strains . Yeast cells could easily be coflocculated at any growth stage . It was concluded that receptor availability is not involved in the process of flocculation onset. Can J Microbiol, 1993 Jan, 39(1), 61 - 9 Changes in the rumen microbial population and its activities during the refaunation period after the reintroduction of ciliate protozoa into the rumen of defaunated sheep; Williams AG et al.; Changes in the microbial populations, their activities, and the ruminal fermentation were monitored for 50 d following the reintroduction of ciliate protozoa into four defaunated sheep . A protozoal population was reestablished successfully in each recipient, using a washed inoculum containing approximately 10(3) cells, although there were between-animal differences in the rates of recolonization and genus establishment . Entodinium spp . predominated in the initial stages of the refaunation period and had an apparent maximal generation time of 9-10 h . Bacterial and fungal numbers did not decline following the reintroduction of protozoa and a small transient increase in the numbers of amylolytic and xylanolytic bacteria and fungal zoospores occurred in the early stages of refaunation when the protozoal population was < 10(5)/g ruminal contents, but these subsequently declined as the protozoa established . Although the fibrolytic bacterial population was lowest in period 3 (> 10(5) protozoa/g), the in sacco ruminal digestion of Lolium perenne hay and polysaccharolytic enzyme activities in the solids-associated populations were either maintained or increased when protozoa were present confirming the important contribution of protozoa to fibre breakdown in the rumen . Significant changes in ruminal microbial activities occurred after protozoal reinoculation but before the rumen had refaunated completely . Arylamidase activities in the liquor-phase population and ruminal ammonia concentrations increased significantly within 48 h of transfaunation; the magnitude of the effects became more pronounced as the protozoal population developed . However, volatile fatty acid formation and ruminal pH were not affected after the reintroduction of protozoa. Arch Microbiol, 1993, 159(2), 174 - 81 Purification of glutaryl-CoA dehydrogenase from Pseudomonas sp., an enzyme involved in the anaerobic degradation of benzoate; Hartel U et al.; Cell-free extracts of Pseudomonas sp . strains KB 740 and K 172 both contained high levels of glutaryl-CoA dehydrogenase when grown anaerobically on benzoate or other aromatic compounds and with nitrate as electron acceptor . These aromatic compounds have in common benzoyl-CoA as the central aromatic intermediate of anaerobic metabolism . The enzymatic activity was almost absent in cells grown aerobically on benzoate regardless whether nitrate was present . Glutaryl-CoA dehydrogenase activity was also detected in cell-free extracts of Rhodopseudomonas, Rhodomicrobium and Rhodocyclus after phototrophic growth on benzoate . Parallel to the induction of glutaryl-CoA dehydrogenase as measured with ferricenium ion as electron acceptor, an about equally high glutaconyl-CoA decarboxylase activity was detected in cell-free extracts . The latter activity was measured with the NAD-dependent assay, as described for the biotin-containing sodium ion pump glutaconyl-CoA decarboxylase from glutamate fermenting bacteria . Glutaryl-CoA dehydrogenase was purified to homogeneity from both Pseudomonas strains . The enzymes catalyse the decarboxylation of glutaconyl-CoA at about the same rate as the oxidative decarboxylation of glutaryl-CoA . The green enzymes are homotetramers (m = 170 kDa) and contain 1 mol FAD per subunit . No inhibition was observed with avidin indicating the absence of biotin . The N-terminal sequences of the enzymes from both strains are similar (65%). Arch Microbiol, 1993, 159(2), 109 - 13 Complete oxidation of benzoate and 4-hydroxybenzoate by a new sulfate-reducing bacterium resembling Desulfoarculus; Drzyzga O et al.; A new sulfate-reducer "strain SAX" was isolated from an anaerobic marine sediment {Saxild, Denmark} . The isolate was a gram-negative, motile and non-spore-forming rod which sometimes appeared as a curved rod . Strain SAX differed from all described Desulfovibrio-, Desulfobotulus- and Desulfoarculus-species by the ability to degrade aromatic compounds such as benzoate, 4-hydroxybenzoate and phenol completely to CO2 . Electron donors used included lactate, pyruvate, malate, fumarate, crotonate and butyrate, while pyruvate was fermented in the absence of an external electron acceptor . Sulfate, thiosulfate or sulfite served as electron acceptors with benzoate as the donor, while nitrate and nitrite did not . The sulfate-reducing bacterium required vitamins and NaCl-concentrations of about 20 g/l . The optimum temperature for growth of strain SAX was 30 degrees C and the optimum pH value was 7.3 . The DNA base composition was 62.4 mol% G+C . The strain possessed cytochrome c3, but no desulfoviridin . On the basis of these characteristics and because strain SAX could not be ascribed to any of the existing species therefore assignment as a new species to the genus Desulfoarculus was suggested. Appl Environ Microbiol, 1993 Jan, 59(1), 290 - 5 Cobalamin-mediated mercury methylation by Desulfovibrio desulfuricans LS; Choi SC et al.; The prominence of sulfate reducers in mercury biomethylation prompted the examination of the methyl carrier and mercury methylation activity of Desulfovibrio desulfuricans LS . There was a low degree of mercury tolerance and a high degree of methylation during fermentative growth; the opposite was true during sulfate reduction . During 2 days of fermentative growth, up to 37% of HgCl2 was methylated at 0.1 micrograms/ml, but only 1.5% was methylated at 10.0 micrograms/ml . Less than 1% of the added HgCl2 was methylated under sulfate-reducing conditions . D . desulfuricans LS radioimmunoassay results were positive for cobalamin . The addition of CoCl2 and benzimidazole to fermentative cultures increased methylation activity . From D . desulfuricans LS grown in the presence of (57)CoCl2, a corrinoid was extracted and purified . High-performance liquid chromatography analysis of the purified extract yielded a single peak with the retention time of cobalamin, and 97% of the (57)Co radioactivity was associated with this peak . Fast atom bombardment and UV and visible spectra of the isolated corrinoid matched those of cobalamin . When methylated with (14)CH3I, the isolated corrinoid methylated Hg(2+) with a 93.9% preservation of (14)C specific activity . We conclude that D . desulfuricans LS methylates mercury via cobalamin (vitamin B12) . Under physiological conditions, the enzymatic catalysis of this reaction is likely. Appl Environ Microbiol, 1993 Jan, 59(1), 255 - 9 Presence of lactate dehydrogenase and lactate racemase in Megasphaera elsdenii grown on glucose or lactate; Hino T et al.; Activity of D-lactate dehydrogenase (D-LDH) was shown not only in cell extracts from Megasphaera elsdenii grown on DL-lactate, but also in cell extracts from glucose-grown cells, although glucose-grown cells contained approximately half as much D-LDH as DL-lactate-grown cells . This indicates that the D-LDH of M . elsdenii is a constitutive enzyme . However, lactate racemase (LR) activity was present in DL-lactate-grown cells, but was not detected in glucose-grown cells, suggesting that LR is induced by lactate . Acetate, propionate, and butyrate were produced similarly from both D- and L-lactate, indicating that LR can be induced by both D- and L-lactate . These results suggest that the primary reason for the inability of M . elsdenii to produce propionate from glucose is that cells fermenting glucose do not synthesize LR, which is induced by lactate. J Dairy Sci, 1993 Jan, 76(1), 245 - 54 Carbohydrate and Aspergillus oryzae effects on intake, digestion, and milk production by dairy cows; Sievert SJ et al.; Six multiparous, ruminally cannulated Holstein cows (46 DIM, 584 kg of BW) and 6 primiparous Holstein cows (35 DIM, 506 kg of BW) were used in two 6 x 6 Latin squares with 21-d periods to examine the effects of level of non-fiber carbohydrate, source of fibrous carbohydrate, and Aspergillus oryzae