Ukr Biokhim Zh, 1994 Jul-Aug, 66(4), 43 - 51 {Formate metabolism by lactate-producing and lactate-utilizing rumen bacteria}; Kalachniuk GI et al.; Physiological concentrations {< 11 mM} of formate do not violate the metabolism of S . bovis and M . elsdenii . A significant inhibition is caused by concentrations of 22 and 44 mM . In this case the process of ammonia formation in S . bovis is inhibited more pronouncedly . Peculiar effects of formate (11 mM) on LDH, FDH, aconitase, isocitrate dehydrogenase, fumarase, L-MDH and malic-enzyme have been stated . The changes show that it enhances assimilation of sugars fermented to lactate in S . bovis, in contrast to M . elsdenii, where it activates the utilization of lactic acid . During the log-phase S . bovis utilized only 11.5% of {14C} H2O2, whereas M . elsdenii uses 33.4% of it . The major amount of the label is transferred from intracellular inclusions to nucleic acids (in S . bovis--74.7%, in M . elsdenii--87%) and then incorporated into low molecular substances (23.5 and 11.9%, respectively), the rest being incorporated into proteins and lipids. Adv Dent Res, 1994 Jul, 8(2), 239 - 45 Saliva stimulation and caries prevention; Edgar WM et al.; The protective role of saliva is demonstrated by the rampant caries seen in human subjects with marked salivary hypofunction, and in desalivated animals . In normal cases, however, the relationship between saliva flow and coronal or root caries experience is doubtful, and to examine the concept that stimulation of saliva might have protective effects against caries, one must look beyond a simple correlation between caries and flow rate . Protective properties of saliva which increase on stimulation include salivary clearance, buffering power, and degree of saturation with respect to tooth mineral . These benefits are maximized when saliva is stimulated after the consumption of fermentable carbohydrates, by reducing the fall in plaque pH leading to demineralization and by increasing the potential for remineralization . Plaque acid production is neutralized, and experimental lesions in enamel are remineralized, when gum is chewed to stimulate saliva after a carbohydrate intake . The pH-raising effects are more easily explained by the buffering action of the stimulated saliva than by clearance of carbohydrates . The remineralization action depends upon the presence of fluoride . These findings suggest that the protective actions of saliva can be mobilized by appropriate salivary stimulation, and that in addition to established procedures such as tooth cleaning and fluoride regimens, eating patterns which lead to saliva stimulation to increase the potential for saliva protection might be included in recommendations for caries prevention . Confirmation of this concept in clinical tests is required. Adv Dent Res, 1994 Jul, 8(2), 221 - 4 Diet patterns and caries; Geddes DA; Few dietary studies have been designed to investigate the effects of intake patterns of food items upon the environment of the teeth . This brief review considers evidence about the effects of choice, combination, and sequence of ingested food and drink upon the pH of human dental plaque in vivo . A series of three studies, which were designed to investigate some of the intra-oral biological events associated with cariogenicity during various eating patterns, are discussed . The principal findings show that if a "meal" includes an item which contains carbohydrate such as sucrose, glucose, or fructose which is rapidly fermented by the acidogenic microorganisms in dental plaque, there will be rapid acid production and the plaque pH will fall . However, other items eaten immediately before, during, or after the consumption of the sugary item can influence the plaque pH . If the non-sugary item stimulates saliva, it will have a pH-raising effect . The remineralizing potential may be enhanced if, for instance, calcium or fluoride is released from the food . However, if one sugary item is followed by another, the demineralizing potential may be enhanced . The results of these experiments are discussed in the context of our current understanding of the dynamics of the carious process . Recent preliminary experiments suggest that other factors, such as the individual subject's speed of consumption, may also affect the cariogenic potential of the oral environment. Biotechnol Prog, 1994 Jul-Aug, 10(4), 428 - 32 A novel ultrasonic resonance field device for the retention of animal cells; Doblhoff-Dier O et al.; This article describes two types of flow-through cell retention devices based on the concept of layered piezoelectric resonators . A single-chamber device is compared to a novel optimized steam-sterilizable prototype ultrasonic cell separator with improved acoustic design and an integrated cooling circuit, eliminating the problem of local temperature increase caused by the high amplitudes necessary to achieve the separation of animal cells with low acoustic contrast . This setup yields highly reproducible results and is ideal for studying the long-term effects of ultrasonic sound fields and separation efficiency . The novel two-chamber system has the potential for scaleability due to the reduction in thermal and acoustic flow, increased field stability, and separation efficiency . Finally, the effect of power input on separation and cell viability is reported . Such flow-through cell retention systems could be used as systems to retain biomass within the fermentor or as a substitute for centrifugation, with the major advantage of eliminating high-speed rotational motion. J Ind Microbiol, 1994 Jul, 13(4), 225 - 32 Glucose and acetate influences on the behavior of the recombinant strain Escherichia coli HB 101 (GAPDH); Gschaedler A et al.; This study highlights data about the production of a recombinant protein (glyceraldehyde-3-phosphate dehydrogenase) by E . coli HB 101 (GAPDH) during batch and fed-batch fermentations in a complex medium . From a small number of experiments, this strain has been characterized in terms of protein production performance and glucose and acetate influences on growth and recombinant protein production . The present results show that this strain is suitable for recombinant protein production, in fed-batch culture 55 g L-1 of biomass and 6 g L-1 of GAPDH are obtained . However this strain, and especially GAPDH overproduction is sensitive to glucose availability . During fermentations, maximum yields of GAPDH production have been obtained in batch experiments for glucose concentration of 10 g L-1, and in fed-batch experiments for glucose availability of 10 g h-1 (initial volume 1.5 L) . The growth of the strain and GAPDH overproduction are also inhibited by acetate . Moreover acetate has been noted as an activator of its own formation. Rev Latinoam Microbiol, 1994 Jul-Sep, 36(3), 177 - 81 {Moraxella bovis biomass production in a bench-top fermentor}; Gonzalez RD et al.; A Moraxella bovis strain was isolated from a kerato-conjunctivities lesion of a calf in Villa Valeria (Cordoba); it was used to establish improved cultural conditions, such as nature and concentration of carbon and nitrogen sources, and pH control in shaken flasks . The selected conditions were assayed for biomass production in a bench-top fermentor . The strain is used by the pharmaceutical industry to produce vaccines and adjuvants . In the initial condition (48 h culture on blood agar) 0.019 g biomass/l.h-1 was obtained . With the use of liquid defined medium with pH control, productivity was increased to 0.153g/l.h-1, with optimum harvest time of 32 h. Mycopathologia, 1994 Jul, 127(1), 19 - 27 Secondary metabolites of Penicillium bilaii strain PB-50; Savard ME et al.; A phosphate-solubilizing strain of Penicillium bilaii was tested for the production of gliotoxin and other toxic compounds . The strain was fermented under five different conditions to allow the expression of various metabolites, including gliotoxin . These included Czapek-yeast extract medium under both shaken and still conditions as well as Czapek-yeast extract/malt extract/peptone medium and sucrose/glycerol medium in shake flasks . In addition, culture filtrate from an industrial fermentation of the fungus was examined . No gliotoxin was produced in any of the media . No other expected P . bilaii metabolites were found . Three compounds were identified in all samples: dibutyl phthalate, 1-(4-hydroxy-phenyl)ethanone and 4-hydroxy-3,6-dimethyl-2H-pyran-2-one . The production of other metabolites was dependent on the culture conditions . Two hyalodendrin derivatives were found in some fermentations and two related compounds were tentatively identified . None of the compounds found have been reported as toxic . The identity of the culture was confirmed by comparison with the ex-type culture of P . bilaii. J Biol Chem, 1994 Jun 17, 269(24), 16726 - 32 Indolepyruvate ferredoxin oxidoreductase from the hyperthermophilic archaeon Pyrococcus furiosus . A new enzyme involved in peptide fermentation; Mai X et al.; Pyrococcus furiosus is a strictly anaerobic archaeon that grows optimally at 100 degrees C by a fermentative-type metabolism in which complex peptide mixtures such as yeast extract and Tryptone, and also certain sugars, are oxidized to organic acids, H2 and CO2 . Enzymes involved in the utilization of peptides such as proteases, aromatic amino transferases, and glutamate dehydrogenase have been previously purified from this organism . It is shown here that P . furiosus also contains significant cytoplasmic concentrations of a new enzyme termed indolepyruvate ferredoxin oxidoreductase (IOR) . This catalyzes the oxidative decarboxylation of aryl pyruvates, which are generated by the transamination of aromatic amino acids, to the corresponding aryl acetyl-CoA . IOR is a tetramer (alpha 2 beta 2) of two identical subunits (66,000 and 23,000 Da) with a molecular weight of 180,000 . The enzyme contains one molecule of thiamine pyrophosphate and four {4Fe-4S}2+,1+ and one {3Fe-4S}0,1+ cluster, as determined by iron analyses and EPR spectroscopy . Significant amounts of other metals such as copper and zinc were not detected . IOR was virtually inactive at 25 degrees C and exhibited optimal activity above 90 degrees C (at pH 8.0) and at pH 8.5-10.5 (at 80 degrees C) . The enzyme was sensitive to inactivation by O2, losing 50% of its activity after exposure to air for 20 min at 23 degrees C, and was quite thermostable, with a half-life of activity at 80 degrees C (under anaerobic conditions) of about 80 min . The Km values (in microM) for indolepyruvate, p-hydroxyphenylpyruvate, phenylpyruvate, CoASH, and P . furiosus ferredoxin, the physiological electron carrier, were 250, 110, 90, 17, and 48, respectively . IOR was inhibited by KCN (apparent Ki = 7.5 mM), but not by CO (1 atm) . An enzyme analogous to IOR has not been reported previously . Curiously, it has few properties in common with the pyruvate ferredoxin oxidoreductase of P . furiosus, even though the two enzymes catalyze virtually identical reactions . In fact, of known ketoacid oxidoreductases, the catalytic mechanism of IOR appears to be most similar to that of the pyruvate ferredoxin oxidoreductase from the hyperthermophilic bacterium Thermotoga maritima. Cancer Res, 1994 Jun 15, 54(12), 3186 - 90 Fermented dairy products, calcium, and colorectal cancer in The Netherlands Cohort Study; Kampman E et al.; Experimental studies suggest that an increased consumption of fermented dairy products and calcium might decrease the risk of colorectal cancer . The associations between fermented dairy products, dietary calcium, and colorectal cancer risk were investigated in a population with a wide variation in intake of dairy products . The Netherlands Cohort Study of diet and cancer started in 1986 when 120,852 Dutch men and women, ages 55-69, filled out a questionnaire concerning dietary patterns and lifestyle . The present analysis is based on 3.3 years of follow-up and includes 215 incident cases of colon cancer and 111 incident cases of rectal cancer, excluding cases diagnosed in the first year of follow-up . After adjustment for potential confounding variables, colorectal cancer risk was weakly inversely associated with the consumption of fermented milk {relative rate (RR) in the highest category of intake compared to nonusers, 0.89; 95% confidence interval (CI), 0.60, 1.33}, unfermented milk (RR, 0.86; 95% CI, 0.57, 1.29), and cheese (RR, 0.88; 95% CI, 0.59, 1.33) . However, category-specific relative rates and tests for trends were not statistically significant . For fermented milk, the inverse association was limited to colon cancer (RR, 0.70; 95% CI, 0.43, 1.15; trend, P = 0.33) . In crude and multivariate models, total dietary calcium intake (highest versus lowest quintile, RR, 0.92; 95% CI, 0.64, 1.34) and calcium from fermented dairy products (RR, 1.14; 95% CI, 0.77, 1.68) were not significantly associated with colorectal cancer risk . Calcium from unfermented dairy products was inversely associated with rectal cancer risk (RR, 0.55; 95% CI, 0.30, 1.04; trend, P = 0.03) . After 3.3 years of follow-up, these data are not consistent with a substantially decreased risk of colorectal cancer with increased intake of fermented dairy products and dietary calcium. J Biol Chem, 1994 Jun 3, 269(22), 15469 - 72 Structure and regulation of SDH3, the yeast gene encoding the cytochrome b560 subunit of respiratory complex II; Daignan-Fornier B et al.; Using an expression library, we have isolated yeast genes activated in the presence of the yeast CCAAT box-binding protein HAP2 . One of these genes, SDH3, encodes the cytochrome b560 subunit of respiratory complex II . The SDH3 protein contains three potential transmembrane domains and is more than 30% identical to bovine cytochrome b560 and to a mitochondrially encoded protein from Marchantia polymorpha . Disruption of SDH3 shows that this gene is required for growth on non-fermentable carbon sources . Expression of SDH1, SDH3, and SDH4 is activated in the presence of the HAP2 transcriptional activator. J Chromatogr B Biomed Appl, 1994 Jun 3, 656(1), 81 - 5 Rapid assay of dinitrophenyl derivative of taurine by high-performance liquid chromatography; Polanuer B et al.; A rapid and simple method for the determination of taurine (2-aminoethanesulphonic acid) in complex samples is described . It is based on the HPLC separation of the dinitrophenyl (DNP) derivative of taurine . The reaction conditions are selected to allow complete derivatization of taurine within 15 min . DNP-taurine samples are stable for at least 3 days . DNP-taurine was separated by reversed-phase liquid chromatography within 12 s . The recovery of taurine was 102 +/- 3% (S.D . = 2.5%, n = 6) and the detection limit was 10 pmol for taurine (signal-to-noise ratio of 10) . The method was applied to the determination of taurine levels in different samples including marine products, infant formulas and fermentation media of different bacterial species. Metabolism, 1994 Jun, 43(6), 728 - 34 Effect of propionate on in vivo carbohydrate metabolism in streptozocin-induced diabetic rats; Cameron-Smith D et al.; Undigested carbohydrates and some dietary fibers are fermented in the large intestine to form short-chain fatty acids (SCFA), including acetate, propionate, and butyrate . It has been suggested that some of the beneficial effects of high-carbohydrate, high-fiber diets on carbohydrate and lipid metabolism are mediated by the metabolism of SCFA in the liver . Propionate has been shown in vitro to decrease glucose production in rat hepatocytes . The aim of the present study was to investigate the effects of propionate on carbohydrate metabolism in normal and streptozocin (STZ)-induced diabetic male Sprague-Dawley rats . Rats were fed a high-fat diet with or without sodium propionate supplementation (either 0.5% or 5% wt/wt) for 4 weeks . At the completion of the feeding period, body weight and liver glycogen concentrations were significantly decreased in STZ-diabetic rats and were unaffected by propionate supplementation . Although STZ-diabetic animals had elevated fasting plasma glucose, cholesterol, and triglyceride levels relative to nondiabetic rats, propionate supplementation had no significant effect on these parameters in either group . Basal and insulin-stimulated carbohydrate metabolism were assessed using the euglycemic clamp technique in overnight-fasted animals with 3(H)-6-glucose infusion . As expected, basal hepatic glucose production (HGP) was higher and the metabolic clearance rate of glucose (MCR) was lower in STZ-diabetic rats . High-dose insulin infusion (3 mU.kg-1.min-1) suppressed HGP in nondiabetic and diabetic animals and increased the MCR in nondiabetic animals . However, propionate supplementation did not alter basal or insulin-stimulated HGP or the MCR in either nondiabetic or diabetic animals.(ABSTRACT TRUNCATED AT 250 WORDS) J Bacteriol, 1994 Jun, 176(11), 3250 - 6 Phosphoenolpyruvate-dependent maltose:phosphotransferase activity in Fusobacterium mortiferum ATCC 25557: specificity, inducibility, and product analysis; Robrish SA et al.; Phosphoenolypyruvate-dependent maltose:phosphotransferase activity was induced in cells of Fusobacterium mortiferum ATCC 25557 during growth on maltose . The disaccharide was rapidly metabolized by washed cells maintained under anaerobic conditions, but fermentation ceased immediately upon exposure of the cell suspension to air . Coincidentally, high levels of a phosphorylated derivative accumulated within the cells . Chemical and enzymatic analyses, in conjunction with data from 1H, 13C, and 31P nuclear magnetic resonance spectroscopy, established the structure of the purified compound as 6-O-phosphoryl-alpha-D-glucopyranosyl-(1-4)-D-glucose (maltose 6-phosphate) . A method for the preparation of substrate amounts of this commercially unavailable disaccharide phosphate is described . Permeabilized cells of F . mortiferum catalyzed the phosphoenolpyruvate-dependent phosphorylation of maltose under aerobic conditions . However, the hydrolysis of maltose 6-phosphate (to glucose 6-phosphate and glucose) by permeabilized cells or cell-free preparations required either an anaerobic environment or addition of dithiothreitol to aerobic reaction mixtures . The first step in dissimilation of the phosphorylated disaccharide appears to be catalyzed by an oxygen-sensitive maltose 6-phosphate hydrolase . Cells of F . mortiferum, grown previously on maltose, fermented a variety of alpha-linked glucosides, including maltose, turanose, palatinose, maltitol, alpha-methylglucoside, trehalose, and isomaltose . Conversely, cells grown on the separate alpha-glucosides also metabolized maltose . For this anaerobic pathogen, we suggest that the maltose:phosphotransferase and maltose 6-phosphate hydrolase catalyze the phosphorylative translocation and cleavage not only of maltose but also of structurally analogous alpha-linked glucosides. J Dairy Sci, 1994 Jun, 77(6), 1630 - 43 Blood and hydrolyzed feather meals as sources of undegradable protein in high fat diets for cows in early lactation; Palmquist DL et al.; Thirty-six cows were in a 2 x 3 factorial study during the first 2 mo of lactation to examine effects on milk yield and composition of added fat (5% of feed DM) and percentage of ruminally undegradable protein (100, 120, or 140% of recommended intake) in the diet . The main source of added undegradable protein was a 1:1 (wt/wt) mixture of blood meal:hydrolyzed feather meal . Diets were low in ADF (ca . 14%) and were highly fermentable in the rumen . The amount of intermediate dietary protein reduced feed intake . Milk yield was high (40 to 44 kg/d), similar among treatment groups, and was sustained for the entire 60-d trail . All cows yielded milk of low fat content (2.1 to 3.2%); supplemental fat decreased proportions of C6 to C14, C18:2, and C18:3 in milk fat and increased C4, C16:0, C18:0, and C18:1 . Higher dietary protein had a positive linear effect on milk fat percentage and increased C16:0 and decreased trans-C18:1 and C18:2 contents of milk fat . Added fat did not change total milk N but increased NPN as a percentage of total milk N . Percentage of total N in milk and yield of whey N was reduced when the intermediate protein diet was fed, associated with the lower DMI of this diet . A requirement for ruminally undegradable protein intake higher than recommended by NRC was not demonstrated with the highly fermentable diets fed in this study; however, ruminal acetate: propionate ratio and milk fat percentage were low. J Dairy Sci, 1994 Jun, 77(6), 1618 - 29 Effects of amount of protein and ruminally protected amino acids in the diet of dairy cows fed supplemental fat; Christensen RA et al.; The objectives of this experiment were to investigate the effects of amount of dietary CP and ruminally protected AA supplementation on production of milk and milk components, ruminal fermentation, and nutrient digestibilities by cows fed diets containing high oil corn and tallow . Holstein cows in midlactation producing 22 to 25 kg/d of milk were used in a 5 x 5 Latin square design . Treatments were 1) control (16.8% CP, no added fat); 2) 14.2% CP, no AA; 3) 14.2% CP, with AA; 4) 17.5% CP, no AA; and 5) 17.5% CP, with AA . Diets 2 to 5 contained supplemental fat from high oil corn and tallow . Diets consisted of 33% alfalfa haylage, 17% corn silage, and 50% concentrate DM . Intake of DM was not different among treatments . Dietary fat increased yields of milk, fat, SNF, and total solids and percentages of fat and total solids . Increasing CP from 14.2 to 17.5% did not alter production or composition of milk . Supplemental AA increased yields of 4% FCM, milk fat, milk CP, true protein, and casein protein and percentages of CP, true protein, and casein protein in milk when either 14.2 or 17.5% CP was in the diet . Supplemental fat did not alter ruminal fermentation, but increases in dietary CP increased total VFA concentration in the rumen without affecting proportions of individual VFA . Apparent digestibilities of DM, OM, CP, starch, and energy in the total tract were greater for cows fed the 17.5% CP diets . Addition of AA to the 14.2% CP diet increased apparent digestibilities of DM, OM, ADF, NDF, and energy in the total tract but decreased digestibilities for cows fed the 17.5% CP diets . Feeding AA to midlactation cows in diets containing supplemental fat may alleviate the decrease in milk protein percentage associated with fat supplementation; this response was similar for cows fed diets that contained either 14.2 or 17.5% CP. J Dairy Sci, 1994 Jun, 77(6), 1589 - 97 Effects of extruded soybeans and forage source on fermentation by rumen microorganisms in continuous culture; Illg DJ et al.; Continuous culture fermenters were used to evaluate effects of extrusion of whole soybeans and changes in forage composition of diets on microbial fermentation . Treatments were arranged in a 2 x 4 factorial design with soybeans (raw or extruded) and dietary treatment (ratio of alfalfa hay to corn silage; 82:18, 61:39, 43:57, and 27:73 of dietary forage) as main effects . Soybeans constituted 9.6, 14.4, 19.2, and 23.9% of DM for each of the respective dietary treatments . True digestion of DM, ADF, and NDF was unaffected by processing of soybeans or dietary treatment, but true digestion of OM decreased as concentration of corn silage and soybeans increased . Total VFA concentration was unaffected by source of soybeans or dietary treatments; however, molar concentration of butyrate decreased in fermenters supplied with diets containing extruded soybeans . Degradation of CP was not influenced by soybean source but decreased as the concentration of corn silage and soybeans increased . Bacterial N output decreased, and dietary N flow from the fermenters increased, as concentration of corn silage and soybeans increased . Changes in the ratio of alfalfa hay to corn silage and alteration of dietary soybean concentration affected true OM digestion and dietary CP degradation, but extrusion of whole soybeans had little effect on fermentation. J Dairy Sci, 1994 Jun, 77(6), 1563 - 9 Influence of corn and sorghum starch on the in vitro kinetics of forage fiber digestion; Grant RJ; In vitro digestion experiments were conducted to examine the interactions of starch source and pH on kinetics and apparent extent of ruminal NDF digestion . Alfalfa hay, bromegrass hay, and each hay with either raw sorghum, raw corn, or pure corn starch in ratios to simulate a 28% NDF diet were incubated at pH 5.5, 6.2, or 6.8 . Ash-free NDF was measured at 0, 6, 12, 18, 24, 30, 36, 48, 72, and 96 h of fermentation; logarithmic transformation and linear regression were used to estimate digestion parameters . Significant forage by starch interactions on lag and rate of NDF digestion and significant forage and starch by pH interactions on rate and potential extent of NDF digestion indicated that forage type must be considered when the effect of pH and starch source on NDF digestion is predicted . For both forages, raw or pure corn starch increased lag more than sorghum starch, especially at low pH . Raw sorghum starch decreased the rate of NDF digestion more for bromegrass hay than for alfalfa hay, and pure corn starch had the largest negative impact on fiber digestion, especially at low pH . Sources of starch influenced lag and rate of NDF digestion differently at pH from 6.8 to 5.5, which led to dramatic differences in apparent extent of ruminal NDF digestion. J Dairy Sci, 1994 Jun, 77(6), 1509 - 14 Quaternary ammonium compounds in milk: detection by reverse-phase high performance liquid chromatography and their effect on starter growth; Valladao M et al.; A reverse-phase HPLC method is described that permits detection of quaternary ammonium sanitizers in milk . The commercial mixture of quaternary ammonium sanitizer used, n-alkyl (50% C14, 40% C12, and 10% C16) dimethyl benzyl ammonium chloride, was extracted from raw and homogenized milks by the Mojonnier milk fat extraction protocol, selectively eluted from the reverse-phase column in the presence of organic solvent and ion-pairing agent, and then analyzed by diode array detection at 217 and 280 nm . Quaternary ammonium sanitizer concentrations in augmented milk samples in the range of 1.0 to 60 micrograms/ml were analyzed with high precision as indicated by a small variance around the mean . Starter culture strains currently in use for manufacture of various fermented milk products available in the Department of Microbiology Culture Collection were examined for their sensitivity to quaternary ammonium sanitizer . All strains were greatly inhibited by as little as 20 micrograms/ml in milk; some were inhibited by only 10 micrograms/ml. J Anim Sci, 1994 Jun, 72(6), 1608 - 15 The influence of intake level and corn processing on digestibility and ruminal metabolism in steers fed all-concentrate diets; Murphy TA et al.; The effects of intake (ad libitum and 70% of ad libitum) and corn processing (whole or rolled) on digestion, ruminal metabolism, and blood metabolites were determined in a 2 x 2 factorial experiment using eight ruminally fistulated steers . All-concentrate diets were fed with adjustments to dietary concentrations of N, vitamins, and minerals to ensure equal daily intake of these nutrients . Digestion of DM, OM, and starch were affected by an intake x processing interaction (P < .03) . Starch digestion for the low-intake, whole-corn diet was much lower than that for the other three diets . This decrease in starch digestion was the major reason for the interaction . Nitrogen digestion was improved (P < .03) with limited intake but was not affected by processing . Ruminal volume and ruminal turnover (percentage/hour) were both reduced (P < .03) on the low-intake diet . Molar proportion of acetate was reduced and propionate was increased (P < .01) for steers receiving high intake of rolled corn compared with the other three diets . Butyrate concentrations were increased (P < .01) when corn was rolled before feeding . Ruminal pH was higher immediately before feeding for steers receiving whole corn than for those receiving rolled corn . Decreases in pH were observed for rolled corn 2 to 9 h after feeding, suggesting a faster rate of fermentation of rolled corn than of whole corn . Concentrations of blood glucose and insulin were not affected by intake or processing . It was concluded that processing corn can improve DM and OM digestibility if intake is restricted and that N digestion is improved with restricted feeding.(ABSTRACT TRUNCATED AT 250 WORDS) Genetics, 1994 Jun, 137(2), 369 - 79 Reduced dosage of genes encoding ribosomal protein S18 suppresses a mitochondrial initiation codon mutation in Saccharomyces cerevisiae; Folley LS et al.; A yeast mitochondrial translation initiation codon mutation affecting the gene for cytochrome oxidase subunit III (COX3) was partially suppressed by a spontaneous nuclear mutation . The suppressor mutation also caused cold-sensitive fermentative growth on glucose medium . Suppression and cold sensitivity resulted from inactivation of the gene product of RPS18A, one of two unlinked genes that code the essential cytoplasmic small subunit ribosomal protein termed S18 in yeast . The two S18 genes differ only by 21 silent substitutions in their exons; both are interrupted by a single intron after the 15th codon . Yeast S18 is homologous to the human S11 (70% identical) and the Escherichia coli S17 (35% identical) ribosomal proteins . This highly conserved family of ribosomal proteins has been implicated in maintenance of translational accuracy and is essential for assembly of the small ribosomal subunit . Characterization of the original rps18a-1 missense mutant and rps18a delta and rps18b delta null mutants revealed that levels of suppression, cold sensitivity and paromomycin sensitivity all varied directly with a limitation of small ribosomal subunits . The rps18a-1 mutant was most affected, followed by rps18a delta then rps18b delta . Mitochondrial mutations that decreased COX3 expression without altering the initiation codon were not suppressed . This allele specificity implicates mitochondrial translation in the mechanism of suppression . We could not detect an epitope-tagged variant of S18 in mitochondria . Thus, it appears that suppression of the mitochondrial translation initiation defect is caused indirectly by reduced levels of cytoplasmic small ribosomal subunits, leading to changes in either cytoplasmic translational accuracy or the relative levels of cytoplasmic translation products. Dtsch Tierarztl Wochenschr, 1994 Jun, 101(6), 237 - 40 {Relationships between postpartum mobilization of body substances and rumen digestion in dairy cows}; Zust J et al.; Relationships between body tissue mobilisation during the postpartal period, and rumen fermentation were studied in 105 high yielding Holstein-Friesian dairy cows . A strongly positive correlation (p < 0.001) between the molar proportion of ruminal acetic acid, and a negative correlation (p < 0.001) between the molar percentage of ruminal propionic and butyric acid (on one hand), and body tissue mobilisation as well as blood plasma concentrations of free fatty acids, beta-hydroxybutyrate, and total bilirubin (on the other hand) were found . These results indicate that in animals with higher postpartal loss of body weight, ruminobacterial fermentation activity is lower, probably secondary to metabolic disturbances. Z Lebensm Unters Forsch, 1994 Jun, 198(6), 480 - 5 Stability of sulphadimidine during raw fermented sausage preparation; Smit LA et al.; The transformation of sulphadimidine (SDM) during raw fermented sausage preparation was studied to elucidate the SDM decrease found in an earlier study . The raw fermented sausages were prepared from batters containing 1 and 10 mg {14C}-SCM kg-1 . The sausages and the brines were analysed using methods based on solid-phase extraction followed by HPLC combined with liquid scintillation counting . It can be concluded that the decrease in SDM level is mainly caused by (i) leaching into the brine (approx . 25%), (ii) transformation of SDM, possibly by reactions with components in the sausage or the brine, as the presence of five reaction products from SDM could be demonstrated, and (iii) formation of bound residues (approx . 20%). J Antibiot (Tokyo), 1994 Jun, 47(6), 639 - 47 Fusarium merismoides Corda NR 6356, the source of the protein kinase C inhibitor, azepinostatin . Taxonomy, yield improvement, fermentation and biological activity; Ohshima S et al.; Fungal strain NR 6356, Fusarium merismoides Corda, was discovered as the source of the protein kinase C (PKC) inhibitor, azepinostatin . The strain was identified based on its growth on potato sucrose agar, slender conidial shape, characteristic polyphialide and production of abundant chlamydospores . Fusarium aquaeductuum Lagh . IMI 103658 and Fusarium sp . NR 7222 were also found to produce the same inhibitor . After single colony isolation and medium optimization trials, a more than 30-fold increase in the production of azepinostatin over the original culture was achieved . Azepinostatin selectively and potently inhibited rat brain PKC with an IC50 value of 70 nM . Other enzymes utilizing ATP, including hexokinase, were not affected . The Ki of azepinostatin for PKC was 0.5 nM . The inhibition of PKC was competitive with ATP and uncompetitive with histone. J Antibiot (Tokyo), 1994 Jun, 47(6), 619 - 30 WS79089A, B and C, new endothelin converting enzyme inhibitors isolated from Streptosporangium roseum . No . 79089 . Taxonomy, fermentation, isolation, physico-chemical properties and biological activities; Tsurumi Y et al.; WS79089A, B and C, which are novel endothelin converting enzyme (ECE) inhibitors have been isolated from the fermentation broth of Streptosporangium roseum No . 79089 . These inhibitors were purified from an acetone extract of whole culture broth followed by Silicar CC-4 column chromatography and HPLC . WS79089A, B and C showed highly selective ECE inhibition activity with IC50 values of 0.73 microM 0.14 microM and 3.42 microM, respectively . On the basis of spectroscopic and chemical evidence, the tentative structures of WS79089A, B and C have been proposed, they have benzo{a}naphtacen chromophores. Rev Sci Tech, 1994 Jun, 13(2), 599 - 614 Traditional methods used for controlling animal diseases in Iran; Tadjbakhsh H; In ancient times in Iran, infectious diseases of animals and human beings were referred to as choleraic diseases . Rhazes (9th century), followed by Avicenna (10th century), Jorjani (11th century) and others, had specific opinions on the cause and effect relationship in these diseases, which recall the fermentation theory of Louis Pasteur . In ancient Iran, the methods adopted for veterinary procedures were those of general theoretical and practical medicine, including the humoral theory, accurate diagnosis, signs and symptoms, and the prescription of herbal and mineral medicines or substances of animal origin . If herbal treatment failed, cauterisation and surgery were used . When refractory and contagious infectious diseases occurred, animals were evacuated from the infected region, in order to preserve their health, with resort to the mercy of Allah (God) as a final remedy . Iranian scientists of ancient times had interesting views on rabies . A kind of serotherapy was used for treating persons bitten by rabid dogs . Vaccination was performed many centuries ago by using dried smallpox lesions . In Baluchistan (Iran), infants were encouraged to play with and touch the teats of cows affected with cowpox, in order to immunise the children against smallpox, and this was centuries before the discovery of smallpox vaccine by Edward Jenner . Camelpox was also used for human immunisation . In the case of caprine pleuropneumonia, an extract or juice was obtained from the lungs of affected animals and was inactivated by treatment with certain herbal medicines which had a disinfectant effect . A thread coated with this extract was passed through the ear of healthy goats to render them immune . The author lists various diseases and their treatment . This work forms part of detailed research by the author with reference to some 2,200 books and many ancient manuscripts on the history of veterinary science in Iran. Br J Nutr, 1994 Jun, 71(6), 871 - 86 Effect of baked beans (Phaseolus vulgaris) on steroid metabolism and non-starch polysaccharide output of hypercholesterolaemic pigs with or without an ileo-rectal anastomosis; Costa NM et al.; The plasma-cholesterol-lowering effects of some dietary legumes are now well established from animal and human studies, but the mechanism is not completely understood . The present study investigated the effect of baked beans (Phaseolus vulgaris) on steroid metabolism of hypercholesterolaemic pigs . Three groups of four pigs were studied: baseline (BL), normal pigs (NP) and those previously prepared with an ileo-rectal anastomosis to nullify the function of the large intestine (IR) . All three groups were given a semi-purified control diet, with about 40% energy as fat (polyunsaturated:saturated fatty acid (P:S) ratio 0.3), supplemented with 10 g cholesterol/kg, for 14 d . Then IR and NP pigs were fed for 28 d on a diet supplemented with 10 g cholesterol/kg and 300 g baked beans/kg (dry-matter basis), so that the 40% contribution to energy from fat was maintained (P:S ratio 0.3) . Group BL was fed on the control diet throughout . The intact pigs (NP) fed on baked beans showed considerable differences compared with the other groups, as follows: (a) reduced plasma cholesterol (NS); (b) higher concentration of cholesterol in bile (NS); (c) higher concentration of bile acids, especially secondary bile acids, in bile (P < 0.05); (d) reduced elimination of bile acids in faeces, especially secondary bile acids (P < 0.05); (e) higher excretion of coprostanol and lower elimination of cholesterol in faeces (P < 0.05) . From these findings it is proposed that a baked-bean-enriched diet potentiates bacterial fermentation and steroid degradation in the large intestine and enhances conservation of bile acids and cholesterol within the enterohepatic circulation . The high concentration of bile acids and cholesterol in bile may thus promote feedback inhibition of hepatic cholesterol synthesis, and hence, reduce plasma cholesterol. Eur J Biochem, 1994 Jun 1, 222(2), 615 - 23 Construction, expression and characterization of a plasmid-encoded Na(+)-specific ATPase hybrid consisting of Propionigenium modestum F0-ATPase and Escherichia coli F1-ATPase; Kaim G et al.; The Escherichia coli strain DK8, a deletion mutant lacking the complete unc operon, was transformed with a plasmid containing the genes encoding the a, b, c, delta and part of the alpha subunit of the Na(+)-dependent ATPase of Propionigenium modestum and the genes encoding the alpha, gamma, beta and epsilon subunits of the H(+)-dependent E . coli ATPase . The transformants showed Na(+)-dependent growth on succinate as non-fermentable carbon source . The functionally expressed hybrid ATPase was activated 13-fold at pH 7.5 by the addition of Na+ and inhibited by 1,3-dicyclohexylcarbodiimide, azide and tributyltin chloride . At pH 7.5 and pH 9.0, the hybrid enzyme was protected from inhibition by 1,3-dicyclohexylcarbodiimide in the presence of 50 mM NaCl and 5 mM NaCl, respectively . The hybrid ATPase was reconstituted into proteoliposomes and catalyzed the transport of Na+ upon ATP addition . ATP-dependent fluorescence quenching of 9-amino-6-chloro-2-methoxyacridine proved that the ATPase hybrid was able to pump protons in the absence of Na+ . Furthermore, ATP synthesis could be measured under conditions where a valinomycin-mediated K+ diffusion potential (delta psi) and a Na+ concentration gradient (delta p Na+) were imposed. Yeast, 1994 Jun, 10(6), 719 - 31 A new nuclear suppressor system for a mitochondrial RNA polymerase mutant identifies an unusual zinc-finger protein and a polyglutamine domain protein in Saccharomyces cerevisiae; Brohl S et al.; A yeast strain with a point mutation in the nuclear gene for the core subunit of mitochondrial RNA polymerase was used to isolate new extragenic suppressors . Spontaneously occurring phenotypical revertants were analysed by crosses with the wild-type and tetrad dissection . One of the new nuclear suppressor mutants was characterized by temperature-sensitive growth on non-fermentable carbon sources . This mutant was transformed with a genomic yeast library . Two independent types of DNA clones were isolated which both complemented the temperature-sensitive defect . Subcloning and DNA sequencing identified two novel yeast genes which code for proteins with the characteristic features of transcription factors . Both factors exhibit highly structured protein domains consisting of runs and clusters of asparagine and glutamine residues . One of the proteins contains in addition zinc-finger domains of the C2H2-type . Therefore the genes are proposed to be named AZF1 (asparagine-rich zinc-finger protein) and PGD1 (polyglutamine domain protein) . Gene disruption of both reading frames has no detectable influence on the vegetative growth on complete glucose or glycerol media, indicating that the genes may act as high copy number suppressors of the mutant defect . Additional transformation experiments showed that AZF1 is also an efficient suppressor for the original defect in the core subunit of mitochondrial RNA polymerase. Voen Med Zh, 1994 Jun, (6), 44 - 5, 80 {The hepatitis C problem in transfusions in military medicine}; Danil'chenko VV et al.; The article contains data on viral hepatitis C, its etiological factor, epidemiology, methods of diagnosis and prophylaxis . It's high time to conduct 100% screening control of donors for viral hepatitis C antibody, using national test system of fermental analysis . The authors worked out the method of "dry drop" to study the serum for hepatitis C virus antibody. Eur J Clin Nutr, 1994 Jun, 48(6), 386 - 96 Dose-response effects of boiled carrots and effects of carrots in lactic acid in mixed meals on glycaemic response and satiety; Gustafsson K et al.; OBJECTIVE: To evaluate the effect of dosage on the metabolic response to vegetables added to a mixed lunch meal, and to relate the amounts to the tripartite plate model . Carrots were chosen as an example, and the carrots were blanched, frozen and boiled to include possible effects of processing and cooking . The effects of carrots steeped in lactic acid, as produced at fermentation, were also studied . DESIGN: The test meals with carrots, and the control meal without vegetables, were balanced regarding energy (2000 kJ) and digestible carbohydrates (60 g) and similar in fat (17 g) and protein (16-19 g) content . The carrot portions of 100, 200 and 300 g contained 2.9, 5.8 and 8.7 g dietary fibre respectively . The meals were served in the morning after an overnight fast and in random order . Blood samples for the analysis of blood glucose, plasma insulin and C-peptide were collected and satiety was graded until 210 min postprandially . SETTING: The study was performed at the research laboratory, Dalby Health Sciences Centre (primary care) . SUBJECTS: The 10 healthy, male volunteers, around 40 years of age, were recruited at random from the district's population list . None dropped out . RESULTS: The larger the carrot portion the lower were the glucose and insulin/C-peptide responses and the higher the satiety scores . The minimum amount causing significant effects was 200 g . According to the plate model, 200 g of boiled carrots was the most that could be included on half the plate . Addition of lactic acid to 200 g carrots augmented the effects on satiety scores and hormonal response . CONCLUSIONS: The addition of generous amounts of vegetables to a mixed meal improves the metabolic response. Clin Invest Med, 1994 Jun, 17(3), 218 - 25 Effect of acute lactulose administration on serum acetate levels in cirrhosis; Fernandes J et al.; Lactulose has been used successfully in the treatment of portal-systemic encephalopathy but its exact mechanism of action is not known . The aim of this study was to observe the systemic effects of the colonic fermentation of an acute lactulose dose in cirrhotics and normal subjects . Six cirrhotic patients and 6 normal subjects were placed on 2 identical 2-d metabolic diets, 1 of which was supplemented with lactulose (1 g/100 kcals to a maximum of 28 g/d) . Lactulose increased colonic fermentation in cirrhotic and normal subjects as evidenced by higher breath hydrogen and serum acetate levels . The increase in serum acetate levels after lactulose compared to control was similar in cirrhotic compared to normal subjects . However, the mean serum acetate concentration in the cirrhotics was significantly greater than that in the control subjects (p = 0.039), indicating increased endogenous production, or decreased peripheral utilization of acetate by the cirrhotic liver, or both . No change was observed in blood ammonia, glucose, insulin, or free fatty acid levels with lactulose. Appl Microbiol Biotechnol, 1994 Jun, 41(4), 373 - 7 Influence of increased dissolved oxygen concentration on productivity and selectivity in cultures of a colabomycin-producing strain of Streptomyces griseoflavus; Dick O et al.; The influence of enhanced O2 concentration on growth and formation of secondary metabolites by Streptomyces griseoflavus (strain Tu 2880) was investigated in a stirred tank and in an air-lift fermentor . At a partial pressure of O2 Po2 = 1880 mbar the growth was lowered by 50% compared to Po2 = 210 mbar, whilst substrate consumption and O2 uptake rate increased markedly . Production of the colabomycin complex reached maximum values at Po2 = 630 mbar . A similar increase of secondary metabolite formation was obtained when glycerol or acetate were fed at Po2 = 220 mbar . The portion of the derivate colabomycin A in the product mixture rose from 43% at Po2 = 210 mbar to 73% at Po2 = 1260 mbar . Since dissolved O2 concentration has a significant influence on productivity and selectivity it may be used to regulate aerobic fermentation processes. Yakugaku Zasshi, 1994 Jun, 114(6), 401 - 13 {Development of bioactive functions in hydrangeae dulcis folium . II . Antiulcer, antiallergy, and cholagoic effects of the extract from hydrangeae dulcis folium}; Yamahara J et al.; In order to develop new bioactive functions of Hydrangeae Dulcis Folium, the fermented and dried leaves of Hydrangea macrophylla Seringe var . thunbergii Makino, effects of the methanolic extract from the crude drug on antiucler, antiallergic, cholagoic, and various pharmacological actions were investigated . Consequently, the methanolic extract was found to exhibit potent antiulcer, antiallergic, and cholagoic activities . By monitoring with these activities, it was found that the active constituents were contained in the lipophilic portion of the methanolic extract . Furthermore, the known lipophilic constituents such as phyllodulcin and hydrangenol were found to show little antiulcer and cholagoic activities, while it was also found that they showed antiallergic activity on Schultz-Dale reactions. Arch Biochem Biophys, 1994 May 15, 311(1), 62 - 71 Regulation of pyruvate carboxylase isozyme (PYC1, PYC2) gene expression in Saccharomyces cerevisiae during fermentative and nonfermentative growth; Brewster NK et al.; In Saccharomyces cerevisiae there are two isoenzymes of pyruvate carboxylase (Pyc) encoded by separate genes, designated PYC1 and PYC2 . In the wild type yeast, the expression of both genes is influenced by both the growth phase and the type of carbon source, indicating discrete regulatory mechanisms and metabolic roles for PYC1 and PYC2 . On glucose minimal medium PYC1 and PYC2 are differentially regulated as shown by a constant level of PYC1 expression throughout the main growth phase compared to a high level of PYC2 expression only in the early growth phase . On ethanol minimal medium, the growth-related pattern of PYC1 and PYC2 expression was similar as shown by a 3.6-fold decline from early to mid log phase . PYC1 expression, however, was activated 10-fold above PYC2 mRNA levels during this period of growth . To further investigate the roles of the two PYC genes we determined the growth phenotypes and expression levels of PYC in pyc1 and pyc2 single null mutants . During fermentative growth, the lack of either PYC gene had little effect on the level and pattern of expression of the other PYC gene, indicating further their separate regulation . In comparison to the pyc2 null, the pyc1 null strain showed a 3- to 4-fold lower level of Pyc activity and Pyc protein concentration . Moreover, the pyc1 null showed a strong requirement for L-aspartate for efficient growth, indicating the importance of PYC1 expression for the synthesis of C4 intermediates . DV6.2 (PYC1, pyc2 delta) showed a 3.2-fold higher level of activity on ethanol minimal medium when compared to growth on glucose minimal medium, and supported growth in the absence of L-aspartate . The pyc1 null, MW21.3 (pyc1 delta, PYC2), on the other hand, did not support growth on ethanol in the absence of aspartate . This study represents the first report on the characterisation of expression of the PYC genes in yeast throughout growth . Their metabolic roles for both fermentative and gluconeogenic growth are considered. FEMS Microbiol Lett, 1994 May 15, 118(3), 213 - 8 Acetaldehyde production in Saccharomyces cerevisiae wine yeasts; Romano P et al.; Eighty-six strains of Saccharomyces cerevisiae were investigated for their ability to produce acetaldehyde in synthetic medium and in grape must . Acetaldehyde production did not differ significantly between the two media, ranging from a few mg/l to about 60 mg/l, and was found to be a strain characteristic . The fermentation temperature of 30 degrees C considerably increased the acetaldehyde produced . This study allowed us to assign the strains to different phenotypes: low, medium and high acetaldehyde producers . The low and high phenotypes differed considerably also in the production of acetic acid, acetoin and higher alcohols and can be useful for studying acetaldehyde production in S . cerevisiae, both from the technological and genetic point of view. Mol Gen Genet, 1994 May 10, 243(3), 358 - 62 Polyubiquitin gene expression contributes to oxidative stress resistance in respiratory yeast (Saccharomyces cerevisiae); Cheng L et al.; UBI4, the polyubiquitin gene of Saccharomyces cerevisiae, is expressed at a low level in vegetative cells, yet induced strongly in response to starvation, cadmium, DNA-damaging agents and heat shock . UBI4 is also expressed at a higher basal level in cells growing by respiration as compared to glucose-repressed cells growing by fermentation . This higher UBI4 expression of respiratory cultures probably helps to counteract the greater oxidative stress of respiratory growth . The effects of inactivating UBI4 on high temperature viability are more marked with respiratory cultures . Also loss of UBI4 leads to a considerably increased rate of killing of respiring cells by hydrogen peroxide, whereas the same gene inactivation has relatively little effect on the peroxide sensitivity of cells in which mitochondrial functions are repressed . This is the first study to reveal that ubiquitin levels in cells can influence their ability to withstand oxidative stress. Leuk Res, 1994 May, 18(5), 319 - 25 Induction of proto-oncogene and cytokine expression in human peripheral blood monocytes and the monocytic cell line THP-1 after stimulation with mycoplasma-derived material MDHM; Quentmeier H et al.; Mycoplasma fermentans-derived high-molecular-weight material (MDHM) was originally described to induce differentiation of murine thymocytes to cytolytic effector T-cells by stimulating IL-6 release from adherent cells . This study shows that human peripheral blood monocytes (PBMo) also respond to MDHM with increases in IL-1 beta, IL-6 and TNF alpha expression, both at the mRNA and protein level . The induced expression of IL-1 beta and TNF alpha mRNA in the monocytic THP-1 cell line increased as quickly as in primary cells . In contrast to PBMo, THP-1 and 14 other monocytic/myeloid leukemia-derived cell lines did not secrete measurable amounts of the cytokines upon treatment with MDHM . IL-1 beta and IL-6 genes contain AP-1 binding sites as regulatory elements, the AP-1 protein being composed of c-jun and c-fos gene products . In THP-1 cells c-jun mRNA expression increased after incubation with MDHM while positive c-fos expression remained unaffected . Although these data suggest AP-1 regulated cytokine mRNA expression, results from PBMo are not in accordance with this notion . In the primary cells MDHM-induced elevation of cytokine mRNA levels was preceded by a downregulation of c-fos expression while positive c-jun expression was not modulated . c-myc mRNA expression, constitutively high in THP-1 cells, was induced in MDHM-stimulated PBMo . In conclusion, MDHM-stimulated induction of cytokine mRNA expression was accompanied by different proto-oncogene responses in PBMo and THP-1 cells . These differences may represent different regulatory pathways of the two cell systems . Alternatively, these data support the notion that neither AP-1 nor the c-myc protein are involved in the MDHM-induced increase in IL-1 beta, IL-6 or TNF alpha mRNA levels . Furthermore, the present results demonstrate clearly that mycoplasma products can have a profound impact on the activation status of eukaryotic cells. Br J Cancer, 1994 May, 69(5), 937 - 42 Starch intake and colorectal cancer risk: an international comparison; Cassidy A et al.; Intakes of starch, non-starch polysaccharides (NSPs), protein and fat have been compared with colorectal cancer incidence in 12 populations worldwide . There were strong inverse associations between starch consumption and large bowel cancer incidence (large bowel r = -0.70, colon r = -0.76) . There was no significant relation with NSPs, although the association with large bowel cancer incidence was still significant when NSP was combined with resistant starch (RS) to give an estimate of fermentable carbohydrate (large bowel r = -0.52, colon r = -0.60) . The relationships between starch, RS and NSPs and cancer incidence remained statistically significant after adjusting for fat and protein intakes . The strong inverse associations found here suggest a potentially important role for starch in protection against colorectal cancer and correspond with the hypothesis that fermentation in the colon is the mechanism for preventing colorectal cancer . Measures of both starch and NSPs need to be included in future epidemiological studies of diet and bowel cancer. Antimicrob Agents Chemother, 1994 May, 38(5), 1123 - 8 Effects of pentamidine isethionate on Saccharomyces cerevisiae; Ludewig G et al.; We used Saccharomyces cerevisiae as a model system in which to examine the mechanism of action of the anti-Pneumocystis drug pentamidine . Pentamidine at low concentrations inhibited S . cerevisiae growth on nonfermentable carbon sources (50% inhibitory concentration {IC50} of 1.25 micrograms/ml in glycerol) . Pentamidine inhibited growth on fermentable energy sources only at much higher concentrations (IC50 of 250 micrograms/ml in glucose) . Inhibition at low pentamidine concentrations in glycerol was due to cytostatic activity rather than cytotoxic or mutagenic activity . Pentamidine also rapidly inhibited respiration by intact yeast cells, although inhibitory concentrations were much higher than those inhibitory to growth (IC50 of 100 micrograms/ml for respiration) . Pentamidine also induced petite mutations, although only at concentrations much higher than those required for growth inhibition . These results suggest that a function essential for respiratory growth is inhibited by pentamidine and that pentamidine affects mitochondrial processes . We propose the hypothesis that the primary cellular target of pentamidine in S . cerevisiae is the mitochondrion. J Anim Sci, 1994 May, 72(5), 1362 - 74 Ruminal digestion and glycosyl linkage patterns of cell wall components from leaf and stem fractions of alfalfa, orchardgrass, and wheat straw; Bourquin LD et al.; Samples of alfalfa, orchardgrass, and wheat straw were hand-separated into leaf and stem fractions that were subjected to in situ ruminal fermentation for various lengths of time to assess the rate and extent of degradation of cell wall neutral monosaccharides, uronic acids, acetyl groups, and hydroxycinnamic acids . A second objective was to measure the glycosyl linkage patterns of leaf and stem fractions of substrates before and after ruminal fermentation . Samples were fermented for 0, 6, 12, 24, 48, 96, and 192 h in each of two ruminally cannulated steers . In situ disappearance data were fitted to a first-order exponential equation to estimate the following substrate parameters: insoluble, potentially digestible fraction (fd), indigestible fraction (fi), and fractional rate constant of degradation of the potentially digestible fraction (k) . Leaves contained larger concentrations of crude protein and smaller concentrations of cell wall components than did stem fractions . Estimates of fi were 7.3, 39.2, 22.1, 49.3, 27.7, and 36.3 for dry matter disappearances for alfalfa leaf, alfalfa stem, orchardgrass leaf, orchardgrass stem, wheat straw leaf, and wheat straw stem, respectively . Averaged across substrates, estimates of fi for arabinose, galactose, glucose, xylose, uronic acids, acetyl groups, and p-coumaric acid were 16.5, 11.4, 14.8, 31.2, 12.8, 25.3, and 22.6% in leaf fractions and 29.5, 19.9, 37.5, 56.2, 35.0, 52.4, and 44.6% in stem fractions . Rates of digestion of all monomeric components except galactose and xylose were greater (P < .05) for alfalfa than for orchardgrass or wheat straw . Differences in digestibility of cell wall components from leaf and stem fractions were greater in alfalfa and orchardgrass than in wheat straw . Glycosyl linkage analysis indicated that xylans in leaf and stem fractions of alfalfa, orchardgrass leaf, and wheat straw stem that resisted degradation had a lower degree of substitution with acid-labile constituents (i.e., other monosaccharides) than was found in original substrates . Different rates and extents of digestion of leaf and stem fractions of forages explain part, but not all, of the observed differences in digestibilities of cell wall monomers by ruminants. Br J Nutr, 1994 May, 71(5), 731 - 7 Breath hydrogen after ingestion of the bulk sweeteners sorbitol, isomalt and sucrose in chocolate; Lee A et al.; The effect of eating chocolate containing sugar alcohols as sweetening agents on colonic fermentation has been investigated by monitoring breath H2 levels . Levels were compared with those occurring after the consumption of normal, sugar-containing chocolate . Ten healthy volunteers aged 19 to 21 years ingested equal amounts of either sorbitol, isomalt or sucrose incorporated into standard chocolate bars . Breath H2 levels after consumption of chocolate containing either sorbitol or isomalt were significantly higher than those after consumption of chocolate containing sucrose (P < 0.001) . After consumption of chocolate containing sorbitol, double the mean estimated volume of breath H2 was produced over 6 h compared with that produced after eating chocolate containing isomalt . Taken together with results relating to the incidence of intolerance symptoms, these findings demonstrate that sorbitol is associated with greater colonic fermentation compared with isomalt. Nutr Rev, 1994 May, 52(5), 176 - 8 Hydroxypropylmethylcellulose, viscosity, and plasma cholesterol control; Topping D; The mechanism for the lowering of plasma cholesterol by water-soluble nonstarch polysaccharides (NSP) could involve alteration of intestinal viscosity leading to attenuated fat and steroid digestion and absorption . Alternatively, there may be direct inhibition of hepatic cholesterol synthesis by short-chain fatty acids produced by large bowel bacterial fermentation . A synthetic NSP, hydroxypropylmethylcellulose (HPMC), has been shown to lower plasma low-density lipoprotein (LDL) cholesterol in humans . This polysaccharide is not fermented by the large bowel microflora and has been shown to lower the plasma and liver cholesterol in hamsters, with no change noted in hepatic sterol synthesis . In further studies with hamsters, a linear relationship has been identified between plasma cholesterol and the logarithm of hydroxymethylcellulose viscosity . Only a relatively small increment in viscosity was necessary to achieve a maximal effect, suggesting that intestinal digestion may be quite sensitive to increased NSP intake. J Clin Microbiol, 1994 May, 32(5), 1387 - 9 Isolation of Mycoplasma species from bronchoalveolar lavages of patients positive and negative for human immunodeficiency virus; Teel LD et al.; The rates of isolation of Mycoplasma species from bronchoalveolar lavages of human immunodeficiency virus (HIV)-infected patients and HIV-negative patients were compared . Mycoplasma species were more frequently isolated from HIV-positive patients . In most cases, a known pulmonary pathogen was also identified . All samples tested negative for Mycoplasma fermentans by PCR. J Dairy Sci, 1994 May, 77(5), 1386 - 98 Comparison of three methods for incorporation of liquid fat into diets for lactating dairy cows; Drackley JK et al.; Two experiments were conducted to determine whether method of incorporation of tallow (iodine value = 57.7) into a TMR for lactating dairy cows affected DMI, milk production or composition, ruminal characteristics, or nutrient digestibilities . In Experiment 1, 8 Holstein and 8 Jersey cows were fed diets containing 1) control, no fat; 2) fat (5% of DM) added first to the concentrate; 3) fat added first to the haylage; and 4) fat added as the last ingredient in the TMR . The DMI was lower when fat was added last to the TMR; DMI was decreased for Jerseys, but not Holsteins, when fat was added first to the concentrate . Milk production was increased, and milk fat percentage decreased, by fat supplementation, but neither differed among application methods . Milk protein percentage was decreased by fat supplementation but was decreased less when fat was added last to the TMR . Production of milk CP, true protein, and casein protein was greater when fat was first mixed with haylage or added last to the TMR . In Experiment 2, four Holstein cows with ruminal cannulas were fed the same diets . Ruminal fermentation characteristics and apparent total tract digestibilities of DM, OM, CP, NDF, ADF, and ash were not different among diets . Digestibility of total fatty acids was decreased when fat was added first to haylage or last to the TMR . Incorporation method had relatively minor effects on variables. J Dairy Sci, 1994 May, 77(5), 1340 - 53 Influence of supplemental protein source and feeding frequency on rumen fermentation and performance in dairy cows; Robinson PH et al.; Multiparous Holstein cows in early lactation were fed a basal mixed ration of 47% (DM) alfalfa and timothy silage and 53% barley and corn concentrate twice daily for ad libitum intake at 1630 and 0600 h . Two supplemental protein sources that differed in their resistance to rumen proteolysis were fed at 9% of total DMI in either two meals per day at 1730 and 0700 h or five meals per day at 1730, 2130, 0200, 0700, and 1200 h . The study was a 4 x 4 Latin square design with six blocks of 4 cows in which one block of cows was fitted with rumen cannulas . Intakes of DM, OM, NDF, and CP were not influenced by treatments . However, cows supplemented with five meals a day tended to consume the mixed ration more rapidly after both the p.m . and a.m . feedings . Milk yield and its content of protein, fat, and lactose also were not influenced by treatments . Average rumen pH was higher, and propionate concentrations were lower, for cows supplemented with five meals, but diurnal patterns were not influenced . Propionate and rumen ammonia N concentrations were lower for cows supplemented with the more resistant protein source; however, rumen VFA, as well as soluble and peptide N concentrations, were not influenced by the type of supplemental protein . Results do not support benefits of synchronized rumen release of energy and N to overall cow production, but rather support previous research that soluble protein or peptide N, or both, may act as a pool to provide N for microbial growth at times of the day when ammonia N concentrations are very low. J Dairy Sci, 1994 May, 77(5), 1167 - 75 A cell culture model to identify biologically active peptides generated by bacterial hydrolysis of casein; MacDonald RS et al.; Consumption of fermented dairy foods has been linked to reduced incidence of colon cancer in population groups . Recently, biologically active compounds have been isolated from these products . Bacterial proteinases, produced by dairy starter cultures, generate a variety of peptides from casein . Some of these casein-derived peptides are likely to alter intestinal cell kinetics . Effects on colon cell kinetics because of the presence of casein-derived peptides may be a mechanism through which fermented dairy foods reduce the risk of colon cancer . We have used two intestinal cell lines (IEC-6 cells, derived from normal rat intestine, and Caco-2 cells, derived from human colon adenocarcinoma) to identify casein peptides that affect intestinal cell kinetics . Cell culture media containing casein were inoculated with three commercial starter cultures and incubated for 4, 8, or 24 h . The bacteria-conditioned media were then filter-sterilized and incubated with the intestinal cells for 6 or 24 h . Rates of {3H}thymidine incorporation and cell cycle kinetics determined by flow cytometry were affected by the culture-modified media in both cell lines . The IEC-6 cells tended to reduce, and Caco-2 cells to increase, rates of cell division after exposure to the media . Intestinal cell response varied among the starter cultures . The results support the use of intestinal cell cultures to identify casein peptides generated by dairy starter cultures, which affect intestinal cell kinetics. J Antibiot (Tokyo), 1994 May, 47(5), 536 - 40 Cytostatin, a novel inhibitor of cell adhesion to components of extracellular matrix produced by Streptomyces sp . MJ654-NF4 . I . Taxonomy, fermentation, isolation and biological activities; Amemiya M et al.; Cytostatin has been identified as a novel inhibitor of cell adhesion to components of extracellular matrix (ECM) in cultured broth of Streptomyces sp . MJ654-NF4 . Though cytostatin did not inhibit EL-4 cell adhesion to ECM components such as laminin and fibronectin; it inhibited the adhesion of B16 melanoma cells to laminin and collagen type IV but not to fibronectin . It exhibited antimetastatic activity on B16 melanoma cells in mice . The cytotoxicity of cytostatin are also reported. J Antibiot (Tokyo), 1994 May, 47(5), 523 - 7 Aselacins, novel compounds that inhibit binding of endothelin to its receptor . I . The producing organism, fermentation and biological activity; Jackson M et al.; A radioligand test to detect inhibitors of endothelin-1 binding to its receptors in bovine atrial and porcine cerebral membranes was used to screen fungal metabolites from stationary fermentations . Inhibitory activity, observed in culture extracts of two Acremonium species, led to the discovery of aselacins A, B and C . Aselacin A inhibits binding to both membrane fractions with IC50s of approximately 20 micrograms/ml. Appl Environ Microbiol, 1994 May, 60(5), 1687 - 9 Degradation of pyrene at low defined oxygen concentrations by a Mycobacterium sp; Fritzsche C; In a fermentor, a Mycobacterium sp . was grown on pyrene at defined oxygen concentrations in a range from 11.4 to 227 microM . The maximal growth rate (mumax = 0.057 h-1) and the dissolved oxygen half-saturation constant (KDO = 5.9 microM) were calculated . At 3.4 microM, the growth rate (mu = 0.011 h-1) was only half of what was expected from the kinetic data . Apparently, this was due to limitation of an oxygenase of pyrene degradation. Appl Environ Microbiol, 1994 May, 60(5), 1519 - 24 Effects of particulate materials and osmoprotectants on very-high-gravity ethanolic fermentation by Saccharomyces cerevisiae; Thomas KC et al.; The effects of osmoprotectants (such as glycine betaine and proline) and particulate materials on the fermentation of very high concentrations of glucose by the brewing strain Saccharomyces cerevisiae (uvarum) NCYC 1324 were studied . The yeast growing at 20 degrees C consumed only 15 g of the sugar per 100 ml from a minimal medium which initially contained 35% (wt/vol) glucose . Supplementing the medium with a mixture of glycine betaine, glycine, and proline increased the amount of sugar fermented to 30.5 g/100 ml . With such supplementation, the viability of the yeast cells was maintained above 80% throughout the fermentation, while it dropped to less than 12% in the unsupplemented controls . Among single additives, glycine was more effective than proline or glycine betaine . On incubating the cultures for 10 days, the viability decreased to only 55% with glycine, while it dropped to 36 and 27%, respectively, with glycine betaine and proline . It is suggested that glycine and proline, known to be poor nitrogen sources for growth, may serve directly or indirectly as osmoprotectants . Nutrients such as tryptone, yeast extract, and a mixture of purine and pyrimidine bases increased the sugar uptake and ethanol production but did not allow the population to maintain the high level of cell viability . While only 43% of the sugar was fermented in unsupplemented medium, the presence of particulate materials such as wheat bran, wheat mash insolubles, alumina, and soy flour increased sugar utilization to 68, 75, 81, and 82%, respectively. FEMS Microbiol Rev, 1994 May, 14(1), 99 - 102 Efficient use of lactose for the lac promoter-controlled overexpression of the main antigenic protein of the foot and mouth disease virus in Escherichia coli under fed-batch fermentation conditions; Neubauer P et al.; Derivatives of the lac promoter (tac, pac, rac) belong to the strongest bacterial promoters which are frequently used for the induced overexpression of foreign genes in Escherichia coli . However, their use in fermentation processes is strongly restricted because of the high cost of the inducer iso-propyl-beta-D-thiogalactopyranoside (IPTG) . The aim of this work was to investigate the possibility of using lac-derived promoters in high cell density processes resulting in a high yield of the induced recombinant protein if glucose is the main carbon and energy source . Lactose is tested as inducer of the main antigenic coat protein (VP1) of the foot and mouth disease (FMD) virus in a T7-RNA polymerase expression system . It was shown that lactose is able to induce the expression of the recombinant gene to an amount of the VP1 protein corresponding to 20% of the total cell protein. FEMS Microbiol Rev, 1994 May, 14(1), 89 - 91 Perfusion systems for hybridoma cells based on sedimentation in chambers and Erlenmeyer flasks; Lassen KM et al.; An inclined sedimentation chamber and a modified 250-ml Erlenmeyer flask have been used as separation devices for perfusion fermentations with hybridoma cells . The maximum cell density is increased 2-16-fold compared to batch fermentations when the separation units are used . When the sedimentation chamber is used, IgG is continuously produced and the daily production is increased by a factor 3.7 compared to batch fermentation. Bull Tokyo Dent Coll, 1994 May, 35(2), 61 - 6 pH response of human dental plaque to chewing gum supplemented with low molecular chitosan; Shibasaki K et al.; The effects of low molecular chitosan (LMCS) on pH responses of human dental plaque following exposure to fermentable carbohydrates were investigated by an ion-sensitive field-effect transistor electrode system . After the plaque pH values were minimized by direct application of 5% glucose solution or consumption of sugared caramel, the subjects started chewing the test gums containing 0 (control), 1 or 3% (w/w) LMCS for three minutes . The pH response was monitored until it recovered to over pH5.5 . In the case of the glucose solution, chewing 3% LMCS gum caused significantly more rapid pH recovery toward the resting level than did the control gum . Initial pH rising rate during gum chewing was faster with either of the two LMCS gums than with the control gum . In the case of caramel, additional effects of LMCS were observed numerically as LMCS content increased . The findings indicated that LMCS had a potential to promote recovery of plaque pH after acidogenic challenge and to maintain the plaque pH around neutrality. Zh Mikrobiol Epidemiol Immunobiol, 1994 May-Jun, (3), 23 - 6 {The heterogeneity of populations of the Escherichia coli recombinant strain KS 1561--a producer of thermolabile enterotoxin during stab cultivation}; Martynenko LD et al.; In the process of the submerged cultivation of E . coli gene engineering strain KS 1561 carrying hybridization plasmid controlling the synthesis of thermolabile enterotoxin LT . The dissociation of the initial strain with the formation of two populations with different properties was observed . One population was characterized by the capacity of fermenting lactose and had decreased activity on the production of LT; the other population, which was lactose-negative, was capable of its increased synthesis. Food Addit Contam, 1994 May-Jun, 11(3), 397 - 402 Investigation of the presence of biogenic amines and ethyl carbamate in kenkey made with maize and maize-cowpea mixtures as influenced by process conditions; Nout MJ et al.; Kenkey is a fermented and cooked maize dough from Ghana . The effect of manufacturing conditions, i.e . fermentation and cooking, and of protein-enrichment by cowpea addition (20% of total weight) on the occurrence of toxic microbial products, namely biogenic amines and ethyl carbamate, were investigated . The levels of biogenic amines in all-maize kenkey were very low (total amines < 60 ppm), but were significantly increased by addition of red cowpea (total amines < 200 ppm, mainly cadaverine and tyramine), and even more by white cowpea (total amines < 500 ppm, mainly putrescine and tyramine) . Histamine was absent (< 5 ppm) in all samples . The effects of fermentation and cooking were less pronounced than the influence of cowpea addition . Prolonged cooking of kenkey resulted in lower levels of putrescine, but did not significantly reduce tyramine levels . Ethyl carbamate levels were negligible (< 11 ppb) in all treatments. Appl Microbiol Biotechnol, 1994 May, 41(3), 317 - 23 Continuous insect cell (Sf-9) culture with aeration through sparging; Wang MY et al.; The continuous growth of Spodoptera frugiperda Sf-9 cells in a 250-ml blown-glass jacketed spinner flask under a direct air sparging environment was investigated . Even at 220 ml working volume (about 90% of total volume), this spinner flask provided good mixing and oxygenation as demonstrated by a higher cell density compared with fermentor cultures . This eliminates a common limitation of the traditional spinner flask, namely much lower cell density at high working volume . Furthermore, this spinner flask has been run with Sf-9 cell culture at five different dilution rates and two different air sparging rates at steady state, demonstrating its utility in research applications where cell size, metabolic activity and environmental conditions can be constantly maintained . In addition to demonstrating the utility of the reactor, three novel points are made in this report . First, cell density in continuous cultures is increased significantly due to a high agitation rate and, especially, air sparging rate, which is seldom used in animal cell or insect cell culture . Second, there is no apparent difference in the specific death rate at two different sparging rates (0.0093 vvm and 0.0125 vvm) . Finally, we have maintained Sf-9 cells for more than 4 months in a continuous culture using a serum-free medium without loss of recombinant protein expression in infected cells. Appl Microbiol Biotechnol, 1994 May, 41(3), 309 - 12 Influence of increased dissolved oxygen concentration on the formation of secondary metabolites by manumycin-producing Streptomyces parvulus; Kaiser D et al.; The influence of increased dissolved O2 concentrations (DOC) on cell growth and production of the secondary metabolite manumycin by a strain of Streptomyces parvulus (Tu 64) was investigated in a stirred tank fermentor . DOC is given as the O2 partial pressure (po2) in the gas phase in an equilibrium state with the liquid phase . Growth of S . parvulus was not influenced up to DOC equivalent to po2 = 1260 mbar . At po2 = 2205 mbar the maximum biomass concentration was lowered by 40% . Production of manumycin was markedly influenced by DOC and reached the maximal concentration at po2 = 315 mbar . At increased DOC three new metabolites were observed . Two of them, 64p-A and 64p-B, were identified as carboxamides, which represent the branched side chain of the manumycin molecule and a derivative with a shorter chain length . The third metabolite, 64p-C, was a manumycin derivative containing an aromatic ring system . Feeding of glycerol during the production phase increased the total yield and showed a similar effect of DOC . Since DOC has significant regulation effects on product formation and selectivity, it should be used as a major parameter in development strategies of aerobic microbial processes. Biotechnol Prog, 1994 May-Jun, 10(3), 308 - 13 Intracellular expression of Vitreoscilla hemoglobin alters the aerobic metabolism of Saccharomyces cerevisiae; Chen W et al.; Vitreoscilla hemoglobin (VHb) has been expressed in Saccharomyces cerevisiae, and its influence on yeast aerobic metabolism has been investigated . New expression vectors were constructed to express VHb constitutively under the control of the ADH-1 promoter . The presence of VHb was shown by Western blot analysis . VHb has been shown to localize predominantly in the cytoplasm . Batch fermentation results indicated that the wild-type strain expressing VHb exhibited a shift in the carbon flux toward ethanol production, with no significant alteration in the specific growth rate . This effect was not observed if cells were grown under respiration inhibition, indicating that the metabolic effect of VHb is likely linked to respiration . Expression of VHb in the adh degrees strain MC65-2A, which produces ethanol only via a respiration-coupled pathway, revealed that ethanol production was decreased and cells reached a higher final cell density in a culture of the VHb-expressing strain . Growth enhancement due to expression of VHb was observed only during the final stage of culture growth when the acetaldehyde produced during the first growth phase was used as a substrate . This metabolic effect of intracellular VHb was seen more clearly in an acetaldehyde fed-batch fermentation in which VHb-expressing cells grew to at least 3-fold higher final cell density . These results suggest that the action of VHb is likely linked to electron transfer. Biotechnology (N Y), 1994 May, 12(5), 494 - 9 Production, purification and immunogenicity of a malaria transmission-blocking vaccine candidate: TBV25H expressed in yeast and purified using nickel-NTA agarose; Kaslow DC et al.; We have constructed a second generation malaria transmission-blocking vaccine candidate based on Pfs25, the predominate surface protein of Plasmodium falciparum zygotes, to overcome potential production problems with the original construct . Four modifications were made: (1) addition of the last cysteine residue of the fourth epidermal growth factor like-domain of Pfs25; (2) mutagenesis of asparagine-linked glycosylation sites with glutamine rather than alanine; (3) addition of a six histidine tag at the carboxy-terminus for highly efficient purification of recombinant protein on nickel-NTA agarose; and (4) fermentation that combines continuous glucose fed-batch methodology with pH-controlled glucose addition and a terminal ethanol feed . The resulting product, TBV25H (Transmission-Blocking Vaccine based on Pfs25 with a Histidine tag), appears to be a more potent antigen and immunogen than the original construct, and the fermentation and post-fermentation processing methodology easily lend themselves to technology transfer to the ultimate users, newly industrialized countries. J Dairy Sci, 1994 Apr, 77(4), 1070 - 83 Effects of soybean hulls and lignosulfonate-treated soybean meal on ruminal fermentation in lactating dairy cows; Mansfield HR et al.; Four Holstein cows were used in a 4 x 4 Latin square design to investigate the effects of soybean hulls and lignosulfonate-treated soybean meal on ruminal fermentation and nutrient passage to the duodenum . Diets contained 32% corn silage, 19.8% alfalfa-grass hay, and 48.2% concentrate (DM basis) . Treatments, arranged in a 2 x 2 factorial, were concentrate mixes based on 1) corn and soybean meal, 2) corn and treated soybean meal, 3) soybean hulls and soybean meal, and 4) soybean hulls and treated soybean meal . Individual protein supplements supplied 40% of dietary CP, and corn or soybean hulls constituted 28% of dietary DM . Intake of OM (mean 18.9 kg/d) was similar among treatments, but intake of NDF was 42% greater, and intake of nonstructural carbohydrate was 55% less, for cows fed soybean hulls . Passage of OM to the duodenum was similar among diets, but flow of NDF was 43% greater, and flow of nonstructural carbohydrate was 56% less, for cows fed soybean hulls . Ruminal pH was similar, but total concentrations of VFA increased 7% when soybean hulls replaced corn . Ruminal digestion of dietary CP was 15% less for cows fed treated soybean meal, but bacterial N flows were similar among treatments . Soybean hulls were digested to a similar extent as corn, but few interactions occurred between supplemental carbohydrate and protein sources. Int J Syst Bacteriol, 1994 Apr, 44(2), 348 - 56 Luteococcus japonicus gen . nov., sp . nov., a new gram-positive coccus with LL-diaminopimelic acid in the cell wall; Tamura T et al.; A new gram-positive, nonmotile coccus is described . Strains IFO 12422T (T = type strain) and IFO 15385 in the Institute for Fermentation, Osaka, culture collection, which were isolated from soil and water, respectively, have the following chemotaxonomic characteristics: menaquinone MK-9(H4); G + C content of DNA of 67 mol%; and LL-diaminopimelic acid, alanine, glycine, and glutamic acid in a molar ratio of ca . 1:2:1:1 (type A3 gamma) . Mycolic acids are not present . The taxonomic characteristics of these organisms are different from those of previously described gram-positive, high-G + C-content cocci . The partial 16S rRNA sequence indicated that IFO 12422T represents a distinct line of descent among gram-positive bacteria with a high G + C content . The name Luteococcus japonicus gen . nov., sp . nov . is proposed . The type strain is strain IFO 12422. Int J Syst Bacteriol, 1994 Apr, 44(2), 324 - 9 Capnocytophaga haemolytica sp . nov . and Capnocytophaga granulosa sp . nov., from human dental plaque; Yamamoto T et al.; Two new Capnocytophaga species, for which we propose the names Capnocytophaga haemolytica and Capnocytophaga granulosa, were isolated from supragingival dental plaque of adults . The phenotypic characteristics of these organisms were the same as those of the genus Capnocytophaga: gram-negative rods; CO2 requirement; gliding motility; catalase negative; oxidase negative; acids produced from D-glucose, D-maltose, D-mannose, and D-sucrose; and acetate and succinate are the major end products of glucose fermentation . In addition, the cellular fatty acid contents and menaquinones of both species were similar to the cellular fatty acid contents and menaquinones of other Capnocytophaga species . Nevertheless, the levels of DNA-DNA relatedness of the two new species to each other and to other Capnocytophaga species were less than 20% . Two notable characteristics of C . haemolytica are the presence of hemolytic activity and the lack of aminopeptidase activity . C . granulosa has granular inclusions in its cells and grows aerobically . The type strains of C . haemolytica and C . granulosa are A0404 (= JCM 8565) and B0611 (= JCM 8566), respectively. FEMS Microbiol Lett, 1994 Apr 1, 117(2), 163 - 8 Pyruvate formate-lyase is not essential for nitrate respiration by Escherichia coli; Kaiser M et al.; Defined deletion mutants of Escherichia coli defective for the synthesis of pyruvate formate-lyase (PFL) or pyruvate dehydrogenase (PDH) were analysed in regards their growth in batch culture and their enzyme levels under fermentative and nitrate respiratory conditions . A pfl mutant proved not to be completely auxotrophic for acetate when grown anaerobically in glucose minimal medium . In contrast, a pfl aceEF double mutant exhibited an absolute requirement for acetate, indicating that PDH is the source of acetyl-CoA in the pfl mutant . Growth of both pfl and aceEF single mutants under nitrate respiratory conditions was essentially indistinguishable from the wild-type . Thus, either PFL or PDH can be used to catabolize pyruvate in nitrate-respiring cells . The activities of PFL and PDH measured after growth with nitrate are commensurate with this proposal. Mol Gen Genet, 1994 Apr, 243(2), 207 - 14 The yeast co-activator GAL11 positively influences transcription of the phosphoglycerate kinase gene, but only when RAP1 is bound to its upstream activation sequence; Stanway CA et al.; Transcription of the yeast phosphoglycerate kinase gene (PGK) is activated by an array of nuclear factors including the multifunctional protein RAP1 . We have demonstrated that the transcriptional co-activator GAL11, which was identified as an auxiliary factor to GAL4 and which is believed to interact with the zinc finger of the trans-activator, positively influences the level of PGK transcription on both fermentable and non-fermentable carbon sources . This positive effect is only observed when the RAP1 site in the upstream activation sequence (UAS) is present, implying that GAL11 acts through RAP1 . Expression of the RAP1 gene is not reduced in the gal11 background, and in vivo footprinting shows that GAL11 does not influence RAP1 DNA-binding activity . Therefore the effect of GAL11 on PGK transcription must be mediated at the PGK UAS, presumably as part of the activation complex . It has been proposed that RAP1 may act as a facilitator of GCR1 binding at the PGK UAS and therefore it is conceivable that the target for GAL11 may in fact be GCR1 . A further implication of this study is that GAL11 can interact with proteins such as RAP1 or GCR1 that are apparently structurally dissimilar from GAL4 and other zinc finger DNA-binding proteins. Am J Clin Nutr, 1994 Apr, 59(4), 879 - 83 Comparative study of the acute effects of resistant starch and dietary fibers on metabolic indexes in men; Ranganathan S et al.; The effect of ingestion of the same amount (30 g) of a resistant starch (lintner) and cellulose on energy expenditure (EE), colonic fermentation (breath-hydrogen test), and blood glucose, insulin, and free fatty acid (FFA) concentrations were compared in seven healthy volunteers in a first experiment . In a second experiment the same indexes were measured in six healthy volunteers after the ingestion of diets composed of 50 g glucose alone or mixed with 30 g lintner, or cellulose, or pectin . In the first experiment no differences between lintner and cellulose were observed on the measured indexes . The notable difference was the increased apparent colonic fermentation with lintner after 6 h . In experiment 2, although insulin response was significantly lower in the pectin-added diet, the results obtained with the four different diets were not significantly different . The metabolic characteristics of lintner were closer to cellulose than to pectin . In conclusion, the acute effect of the ingestion of a resistant starch (lintner) on the measured metabolic indexes is similar to that of a known insoluble fiber--cellulose. Mol Cell Biol, 1994 Apr, 14(4), 2740 - 54 Identification and characterization of a novel yeast gene: the YGP1 gene product is a highly glycosylated secreted protein that is synthesized in response to nutrient limitation; Destruelle M et al.; Nutrient starvation in the yeast Saccharomyces cerevisiae leads to a number of physiological changes that accompany entry into stationary phase . The expression of genes whose products play a role in stress adaptation is regulated in a manner that allows the cell to sense and respond to changing environmental conditions . We have identified a novel yeast gene, YGP1, that displays homology to the sporulation-specific SPS100 gene . The expression of YGP1 is regulated by nutrient availability . The gene is expressed at a basal level during "respiro-fermentative" (logarithmic) growth . When the glucose concentration in the medium falls below 1%, the YGP1 gene is derepressed and the gene product, gp37, is synthesized at levels up to 50-fold above the basal level . The glucose-sensing mechanism is independent of the SNF1 pathway and does not operate when cells are directly shifted to a low glucose concentration . The expression of YGP1 also responds to the depletion of nitrogen and phosphate, indicating a general response to nutrient deprivation . These results suggest that the YGP1 gene product may be involved in cellular adaptations prior to stationary phase and may be a useful marker protein for monitoring early events associated with the stress response. J Bioenerg Biomembr, 1994 Apr, 26(2), 157 - 65 Intermediate metabolism in Trypanosoma cruzi; Cazzulo JJ; Epimastigotes of Trypanosoma cruzi, the causative agent of Chagas disease, catabolize proteins and amino acids with production of MH3, and glucose with production of reduced catabolites, chiefly succinate and L-alanine, even under aerobic conditions . This "aerobic fermentation of glucose" is probably due to both the presence of low levels of some cytochromes, causing a relative inefficiency of the respiratory chain for NADH, reoxidation during active glucose catabolism, and the lack of NADH dehydrogenase and phosphorylation site I, resulting in the entry of reduction equivalents into the chain mostly as succinate . Phosphoenol pyruvate carboxykinase and pyruvate kinase may play an essential role in diverting glucose carbon to succinate or L-alanine, and L-malate seems to be the major metabolite for the transport of glucose carbon and reduction equivalents between glycosome and mitochondrion . The parasite contains proteinase and peptidase activities . The major lysosomal cysteine proteinase, cruzipain, has been characterized in considerable detail, and might be involved in the host/parasite relationship, in addition to its obvious role in parasite nutrition . Among the enzymes of amino acid catabolism, two glutamate dehydrogenases (one NADP- and the other NAD-linked), alanine aminotransferase, and the major enzymes of aromatic amino acid catabolism (tyrosine aminotransferase and aromatic alpha-hydroxy acid dehydrogenase), have been characterized and proposed to be involved in the reoxidation of glycolytic NADH. Protein Expr Purif, 1994 Apr, 5(2), 192 - 7 Expression of human placental alkaline phosphatase in Escherichia coli; Beck R et al.; The human placental alkaline phosphatase gene was subcloned for expression in Escherichia coli . A shortened form of the alkaline phosphatase, lacking 29 C-terminal amino acids which constitute a membrane anchoring domain, is expressed to about 5% of total protein . Most of the enzyme is present in an insoluble form; however, soluble enzyme can be detected in Western blots and activity tests . The protein is located in the periplasm since its signal peptide is cleaved off . Like the wild-type enzyme, recombinant, shortened placental alkaline phosphatase is inhibited by L-phenylalanine . The amount of active enzyme can be increased by addition of magnesium but not zinc to the medium . Using optimized fermentation conditions, 11 kU of soluble and active alkaline phosphatase are produced per liter . Coexpression of potential folding aids like peptidyl-prolyl cis-trans isomerase or disulfide isomerase does not lead to an increase of soluble enzyme, either when overexpressed separately or from an operon. Protein Expr Purif, 1994 Apr, 5(2), 105 - 11 Preparative scale culture of Escherichia coli cells expressing the human immunodeficiency virus type 1 Tat protein; McKenna MC et al.; A procedure leading to a 100-liter fermentor culture of Escherichia coli cells expressing the human immunodeficiency virus type 1 (HIV-1) trans-activator (Tat) protein is described . The effects of growth temperature and of cell density at the time of induction on the yield of Tat were investigated . Tat was identified by SDS-gel electrophoresis and Western blot . Tat represents approximately 10% of the soluble protein in the cell lysate. Scand J Gastroenterol, 1994 Apr, 29(4), 309 - 12 Acetaldehyde and ethanol production by Helicobacter pylori; Salmela KS et al.; By virtue of possessing alcohol dehydrogenase activity, cytosol prepared from Helicobacter pylori produces toxic acetaldehyde from ethanol in vitro . To approach the in vivo situation in the stomach, we have now investigation whether intact H . pylori--without addition of exogenous nicotinamide adenine dinucleotide--also forms acetaldehyde . Furthermore, to assess the energy metabolism of H . pylori, we determined whether the alcohol dehydrogenase-catalyzed reaction can run in the opposite direction with ethanol as the end-product and thereby yield energy for the organism . Intact H . pylori formed acetaldehyde already at low ethanol concentrations (at 0.5% ethanol, acetaldehyde, 64 +/- 21 and 75 +/- 9 mumol/l (mean +/- SEM) for strains NCTC 11637 and NCTC 11638, respectively) . H . pylori produced ethanol in concentrations that can be significant for the energy metabolism of the organism . Acetaldehyde production by H . pylori may be an important factor in the pathogenesis of gastroduodenal diseases associated with the organism . The primary function of H . pylori alcohol dehydrogenase may, however, be alcoholic fermentation and consequent energy production under microaerobic conditions. J Clin Microbiol, 1994 Apr, 32(4), 897 - 902 Hybridization of strains of Escherichia coli O157 with probes derived from the eaeA gene of enteropathogenic E . coli and the eaeA homolog from a Vero cytotoxin-producing strain of E . coli O157; Willshaw GA et al.; A total of 375 Escherichia coli O157 strains were tested by colony hybridization with the eae probe from the central portion of the eaeA gene of the classical enteropathogenic E . coli strain E2348/69 . They were also tested with a probe, eaeO157, from the C-terminal end of the eae gene homolog from a Vero cytotoxin (VT)-producing strain of E . coli (VTEC) of serotype O157:H7 . Both probes hybridized with all 246 O157:H7 or H- VTEC strains tested . The majority were from human infections, and the remainder were from cattle . A further 10 strains (H7 or H-) hybridized with both eae and eaeO157 sequences but not with VT probes . They resembled O157 VTEC and were probably naturally occurring derivatives that had lost VT genes . The remaining 119 strains of O157 were from human, animal, and food sources and belonged to 16 H types other than H7 or were H- . They were VT negative and differed in their properties from O157 VTEC: generally they fermented sorbitol in 1 day, produced beta-glucuronidase, and could not be phage typed by the scheme for O157 VTEC . The eae probe but not the eaeO157 sequence hybridized with 18 H8 or H39 strains, predominantly from human diarrhea . The remaining 101 VT-negative strains hybridized with neither probe . However, 16 strains of O157:H45 hybridized with a probe for diffusely adherent E . coli and attached to HEp-2 cells in a diffuse pattern . Serogroup O157 comprises strains with heterogeneous properties . The eaeO157 probe is a valuable addition to the VT probes used to differentiate O157 strains. J Clin Microbiol, 1994 Apr, 32(4), 1101 - 3 Antibiotic susceptibilities of AIDS-associated mycoplasmas; Poulin SA et al.; Because mycoplasmas may be a cofactor in the progression of human immunodeficiency virus infection to AIDS, their susceptibilities to antibiotics need to be known in the event that appropriate therapy is required . The mycoplasmas studied were a stock culture strain of Mycoplasma fermentans, two strains of M . fermentans isolated from patients with AIDS, M . fermentans var . incognitus, Mycoplasma penetrans, and Mycoplasma pirum . The antibiotics tested were doxycycline, tetracycline, clindamycin, ofloxacin, erythromycin, azithromycin, and clarithromycin at levels consistent with the attainable levels in serum . By the macrodilution metabolic inhibition method, all six mycoplasma strains were susceptible to doxycycline, tetracycline, clindamycin, ofloxacin, azithromycin, and clarithromycin . M . penetrans was susceptible to erythromycin . The M . fermentans strains and M . pirum were resistant to erythromycin . The macrodilution metabolic inhibition method results showed agreement with the Sensititre Gram Positive MIC Panel results for tetracycline, clindamycin, and erythromycin . MICs of clarithromycin for all six mycoplasma isolates tested were low, indicating susceptibility. J Nat Prod, 1994 Apr, 57(4), 541 - 4 Secondary metabolites by chemical screening, 26.1 7-O-beta-D-galactosyl-brefeldin A via transglycosylation with Penicillium brefeldianum; Grabley S et al.; A lactose-containing fermentation medium initiated the formation of 7-O-beta-D-galactosyl-brefeldin A {1} in the brefeldin A-producing organism, Penicillium brefeldianum . The detection, isolation, and physicochemical properties of 1, which has been produced by biological derivatization via transglycosylation, are described . Compound 1 is the first glycosylated brefeldin A derivative to have been prepared. Appl Environ Microbiol, 1994 Apr, 60(4), 1087 - 92 Pentose utilization by the ruminal bacterium Ruminococcus albus; Thurston B et al.; Ruminococcus albus is an important fibrolytic ruminal bacteria which degrades hemicellulose and ferments the resulting pentose sugars . However, little information is available on the utilization of pentoses by this organism or the effect of hexose sugars on pentose metabolism . Enzymatic studies indicated that R . albus metabolized pentoses via the pentose phosphate pathway and possessed constitutive transketolase activity . Cellobiose was preferred over xylose and arabinose, and it appeared that the disaccharide decreased pentose metabolism by repression of transport activity and catabolic enzymes (isomerases and kinases) . Glucose and xylose were co-utilized, and transport studies suggested that there was a common transport system for both sugars . In contrast, glucose was preferred over arabinose and the hexose noncompetitively inhibited the transport of arabinose . Since R . albus lacks a glucose phosphotransferase system, the inhibition of arabinose uptake could not be explained by previously described models of inducer exclusion involving such a system . Because accumulation of radiolabeled xylose, arabinose, and glucose proceeded in the absence of a proton motive force and since transport was correlated with the intracellular ATP concentration, it appeared that monosaccharide uptake was driven by ATP hydrolysis. J Anim Sci, 1994 Apr, 72(4), 1038 - 42 Influence of yucca extract on ruminal metabolism in cows; Wu Z et al.; Two trials were conducted to determine the influence of yucca extract on ruminal digestion, fermentation, and ammonia patterns using ruminally and duodenally cannulated dairy cows . In Trial 1, urea at 0 or 1% of the diet and yucca extract at 0 or 4 g/d formed four dietary treatments in a 2 x 2 factorial arrangement . The experimental design was a 4 x 4 Latin square with 15-d periods . Duodenal digesta were sampled every 6 h during the last 4 d of each period to determine OM and ADF digestibilities and bacterial protein synthesis in the rumen using Cr2O3 and 15N markers . Ruminal digestibilities were (percentage): OM 46.3 vs 43.0%, and ADF 35.9 vs 41.4%, with or without Deodorase . Microbial protein entering the duodenum averaged 2.7 vs 3.1 kg/d for the respective treatments . Ruminal measurements were not affected by treatment (P > .10) . In Trial 2, five cows were used in a 5 x 5 Latin square with 7-d periods . Treatments were 0, 2, 4, 6, and 8 g/d of yucca extract administered via ruminal cannulas . Ruminal fluid was sampled 0, 1, 2, 4, 7, 11, 16, and 22 h after feeding during the last 2 d of each period . Average ruminal NH3 N ranged from 31.4 to 35.4 mg/dL, pH 5.99 to 6.18, and total VFA from 120 to 129 mM, and all did not differ among treatments (P > .10) . Yucca extract administered at 4 g/d did not significantly affect ruminal digestibilities of OM and ADF, and up to 8 g/d did not affect ruminal NH3, pH, or VFA. Mol Biochem Parasitol, 1994 Apr, 64(2), 253 - 60 Entamoeba histolytica has an alcohol dehydrogenase homologous to the multifunctional adhE gene product of Escherichia coli; Yang W et al.; Entamoeba histolytica ferments glucose to ethanol under the anaerobic conditions of the human colon . There is special interest in this metabolic pathway because it provides an opportunity for parasite-specific chemotherapy . Peptide sequences from a 97-kDa E . histolytica protein, which was originally isolated because of extracellular matrix binding properties, were used to clone and sequence a gene that was found to encode an E . histolytica alcohol dehydrogenase and acetaldehyde dehydrogenase (EhADH2) . The EhADH2 cDNA clone had an open reading frame encoding 870 amino acids with a predicted molecular weight of 95,758 . The EhADH2 cDNA clone was identical in 48% of its amino acids to the multifunctional enzyme (alcohol dehydrogenase, acetyl-CoA reductase, and pyruvate-formate-lyase-deactivase) encoded by the Escherichia coli adhE gene . The isolation of the EhADH2 protein helps define a new family of ADH enzymes that may be specific to anaerobic and facultatively anaerobic organisms. Antibiot Khimioter, 1994 Apr, 39(4), 16 - 22 {Sorption of aminoglycoside antibiotics from unfiltered culture fluid}; Bulycheva MS et al.; Sorption of aminoglycosides such as sisomicin, gentamicin, kanamycin and streptomycin from the non-filtrated fermentation broth by carboxylic cation exchange resins under the static conditions was studied . The optimal conditions for the sorption of the aminoglycosides and their subsequent desorption were determined (the ratio of the volumes of the sorbent and fermentation broth and the time of the sorption and desorption). Bioseparation, 1994 Apr, 4(2), 89 - 99 E . coli penicillin acylase isolation by selective release, aqueous two-phase partitioning and ultrafiltration; Guan Y et al.; A procedure has been developed for the purification of penicillin acylase from E . coli fermentation broths . The method described is based primarily on selective release from biomass, charge-directed partitioning in an aqueous two-phase system, and the use of ultrafiltration membranes to recycle the modified polyethylene glycol required and to further purify the enzyme . Each step has been discussed and comparisons have been made with other approaches, where possible . Suggestions have been made for optimisation in process engineering . The approach developed may be applicable to other beta-lactam antibiotic acylases or more generally to some of the other E . coli periplasmic proteins. Biosci Biotechnol Biochem, 1994 Apr, 58(4), 627 - 30 High level expression of Fusarium alkaline protease gene in Acremonium chrysogenum; Morita S et al.; We transformed Acremonium chrysogenum with the genomic DNA of the alkaline protease (Alp) from Fusarium sp . S-19-5 including its promoter . Most of the transformants thus obtained produced a large amount of Alp . PCR and Southern hybridization analysis of genomic DNAs from these transformants showed chromosomal integration of the full-length Alp gene . SDS-PAGE analysis of the supernatant from the transformants showed the presence of Fusarium Alp . The amino terminus of the Alp produced in A . chrysogenum was identical to that of native Fusarium Alp . These results indicate that the Alp promoter, signal sequence, and introns functioned correctly in A . chrysogenum . One of the transformants produced more than 4 g of the Alp per liter in a jar fermentor. J Antibiot (Tokyo), 1994 Apr, 47(4), 391 - 8 WIN 64821, a novel neurokinin antagonist produced by an Aspergillus sp . I . Fermentation and isolation; Sedlock DM et al.; WIN 64821, a nonpeptide neurokinin antagonist, was isolated from a strain of Aspergillus sp., SC319 . The compound was produced in different fermentation media with greatest yields observed when the culture was grown in a synthetic medium supplemented with L-tryptophan and L-phenylalanine . After 6 days fermentation, yields greater than 600 mg/liter were obtained . Two analogs of WIN 64821 were also identified in the culture extracts and subsequently tested for biological activity . WIN 64821 was the most potent compound isolated from this culture and exhibited activity as a substance P-binding inhibitor with submicromolar potency against the human neurokinin 1 receptor. Brain Res, 1994 Mar 28, 641(1), 51 - 6 Mycoplasma triggering of nitric oxide production by central nervous system glial cells and its inhibition by glucocorticoids; Brenner T et al.; The same cytokines that have been implicated in the pathology of central nervous system (CNS) inflammatory diseases and demyelinating diseases are also associated with the induction of nitric oxide (NO) production by macrophages and other somatic cells . Recently we have showed that mycoplasma can trigger the production of tumor necrosis factor (TNF)alpha and eicosanoids in rat astrocytes . In the present study, the effect of mycoplasma on NO production in rat glial cells was assessed . The addition of 10 micrograms/ml of membranes derived from M . capricolum (sheep isolate), M . fermentans (human isolate), or lipopolysaccharide (LPS) led to a 15- to 20-fold increase in NO production . The glucocorticoids dexamethasone and corticosterone, but not progesterone, markedly inhibited NO production . The addition of glucocorticoid prior or conjointly with the activator prevented large amounts of NO from being formed . Even when glucocorticoids were added 5 or 24 h after activation, effective inhibition of NO production was obtained . Thus, it is likely that glucocorticoids exert some of their ameliorating effects in neurological diseases by reducing the production of NO, cytokines and prostaglandins in the CNS. J Biol Chem, 1994 Mar 25, 269(12), 9045 - 51 Deletion of the receptor MOM19 strongly impairs import of cleavable preproteins into Saccharomyces cerevisiae mitochondria; Moczko M et al.; The mitochondrial outer membrane proteins MOM19 and MOM72 are thought to function as import receptors for nuclear encoded preproteins . Different views exist about the importance of each receptor in the import of cleavable and noncleavable preproteins into mitochondria . Here we cloned and sequenced MOM19 from Saccharomyces cerevisiae and constructed a gene disruption mutant . Yeast cells lacking MOM19 were unable to grow on nonfermentable carbon sources and were slow in growing on a fermentable medium, while the growth of yeast cells lacking MOM72 (Mas70p) was much less impaired . delta MOM19 cells accumulated considerable amounts of mitochondrial preproteins in vivo . The import of cleavable preproteins into isolated delta MOM19 mitochondria was strongly inhibited, while import of the noncleavable ADP/ATP carrier and phosphate carrier was only slightly inhibited . The reciprocal situation was found for protein import into delta MOM72 mitochondria . In particular, import of the cleavable precursor of cytochrome c1 into delta MOM72 mitochondria was, in agreement with a previous report (Hines, V., and Schatz, G . (1993) J . Biol . Chem . 268, 449-454), found to be partially inhibited, yet a much stronger inhibition of import was seen into delta MOM19 mitochondria . The direct comparison of protein import into yeast mutants of either receptor yields a unifying hypothesis on mitochondrial preprotein targeting; both receptors have an overlapping specificity, and MOM19 plays a major role for cleavable preproteins . Interestingly, the primary sequence of MOM19 predicts the presence of a tetratricopeptide motif that was also found in MOM72, in the peroxisomal membrane protein PAS8/PAS10, and in several proteins involved in RNA synthesis or mitosis. Biochem J, 1994 Mar 15, 298 Pt 3, 719 - 25 Periplasmic expression of human interferon-alpha 2c in Escherichia coli results in a correctly folded molecule; Voss T et al.; Human interferon-alpha 2c (IFN-alpha 2c) was produced in Escherichia coli under the control of the alkaline phosphatase promoter using a periplasmic expression system . Compared with other leader sequences, the heat-stable enterotoxin II leader of E . coli (STII) resulted in the highest rate of correct processing as judged by Western-blot analysis . The fermentation was designed as a batch-fed process in order to obtain a high yield of biomass . The processing rate of IFN-alpha 2c could be increased from 25% to more than 50% by shifting the fermentation pH from 7.0 to 6.7 . IFN-alpha 2c extracted from the periplasm was purified by a new four-step chromatographic procedure . Whereas cytoplasmically produced IFN-alpha 2c does not have its full native structure, IFN-alpha 2c extracted from the periplasm was found to be correctly folded, as shown by c.d . spectroscopy . Peptide-map analysis in combination with m.s . revealed the correct formation of disulphide bridges . N-terminal sequence analysis showed complete removal of the leader sequence, creating the authentic N-terminus starting with cysteine. J Biol Chem, 1994 Mar 4, 269(9), 6664 - 70 Transcriptional modulation by n-butyric acid of beta 1-, beta 2-, and beta 3-adrenergic receptor balance in 3T3-F442A adipocytes; Krief S et al.; 3T3-F442A adipocytes, which express major beta 3-adrenergic receptors (beta 3-AR) (90%) and minor beta 1-AR (< 10%) and beta 2-AR (< 1%) populations, were used to investigate regulation by n-butyric acid of beta-AR subtype expression . Following butyrate treatment, EC50 values of beta 1- and beta 2-selective agonists, dobutamine and fenoterol, were decreased, whereas that of the beta 3-selective agonist BRL37344 was increased . Direct binding and competition of (-)-{125I}iodocyanopindolol binding by selective beta 1- and beta 2-AR antagonists, CGP20712A and ICI118551, and by the beta 3-AR agonist, BRL37344, revealed that both beta 1- and beta 2-AR were increased in butyrate-treated adipocytes, whereas beta 3-AR almost totally disappeared . In control adipocytes, beta 1-, beta 2-, and beta 3-AR transcripts (quantitated by a polymerase chain reaction assay) represented 6.5, 0.5, and 93% of total beta-AR mRNA, respectively . In butyrate-exposed cells, proportions of beta-AR proteins and mRNAs were, respectively, 87 and 94% for beta 1 and 9 and 1% for beta 2-AR . beta 3-ARs were barely detectable in binding assays and accounted for 4.5% of beta-AR transcripts . Variations of beta-AR protein and mRNA levels were accompanied by parallel changes in the transcription rates of the corresponding genes . The differential regulation of the three beta-ARs by n-butyric acid, a dietary factor produced from colonic fermentation, may have significant nutritional and energetic consequences. Blutalkohol, 1994 Mar, 31(2), 76 - 9 {Ethanol content of Kefir water}; Rabl W et al.; The question of the influence of kefir on blood-alcohol-level has been asked in a legal proceeding . The questioned recipe consisted of 21 water, 6 soup-spoons of kefir granules (about 120 g), 150 g sugar, 2 figs and one lemon . The consumption took place after two days of fermentation . Experimentally we found, that one liter of this kefir product may contain up to 38 g/l ethanol after 7 to 10 days . On the second day we measured up to 16 g/l ethanol . Our results may be import for expert appraisements concerning unability of driving. J Nat Prod, 1994 Mar, 57(3), 363 - 8 New teleocidin-related metabolites, (-)-7-geranylindolactam V and blastmycetin F, from Streptoverticillium blastmyceticum; Irie K et al.; Two new teleocidin-related metabolites, (-)-7-geranylindolactam-V {2} and blastmycetin F {3}, were isolated from fermentation broths of the actinomyce