J Agric Food Chem, 2000 Apr, 48(4), 1377 - 83 Occurrence of Fusarium species and mycotoxins in nepalese maize and wheat and the effect of traditional processing methods on mycotoxin levels; Desjardins AE et al.; Maize (Zea mays) and wheat (Triticum aestivum) collected in the foothills of the Nepal Himalaya Mountains were analyzed for Fusarium species and mycotoxins: fumonisins, nivalenol (NIV), and deoxynivalenol (DON) . Predominant species were Gibberella fujikuroi mating population A (F . moniliforme) in maize and F . graminearum in maize and wheat; G . fujikuroi mating population D (F . proliferatum), F . acuminatum, F . avenaceum, F . chlamydosporum, F . equiseti, F . oxysporum, F . semitectum, and F . torulosum were also present . Strains of G . fujikuroi mating population A produced fumonisins, and strains of F . graminearum produced NIV or DON . By immunoassay or high-performance liquid chromatography, fumonisins were >1000 ng/g in 22% of 74 maize samples . By immunoassay or fluorometry, NIV and DON were >1000 ng/g in 16% of maize samples but were not detected in wheat . Fumonisins and DON were not eliminated by traditional fermentation for producing maize beer, but Nepalese rural and urban women were able to detoxify contaminated maize by hand-sorting visibly diseased kernels. J Agric Food Chem, 2000 Apr, 48(4), 1367 - 72 Soybean meal fermented by Aspergillus awamori increases the cytochrome P-450 content of the liver microsomes of mice; Kishida T et al.; The effect of soybean meal fermented by Aspergillus awamori on the acute lethality of acetaldehyde, pentobarbital sleeping time, and cytochrome P-450 content of the hepatic microsomes was studied in mice . Most of the daidzin and genistin in soybean meal (SBM) were converted into the respective aglycones, daidzein and genistein, by fermentation . In experiment 1, mice were fed isonitrogenic test diets with one of the following five protein sources for 28 d: casein, SBM, fermented and hot-air-dried SBM (FSBM-HD), fermented and freeze-dried SBM (FSBM-FD), or methanol-extracted FSBM-FD (FSMB-FD-R) . The acute lethality of acetaldehyde in mice fed the FSBM-FD diet was significantly lower than that in mice fed the SBM, FSBM-HD, or FSBM-FD-R diet . In experiments 2 and 3, mice were fed isonitrogenic test diets with one of the following four protein sources for 28 d: casein, SBM, FSBM-FD, and FSBM-FD-R . The pentobarbital sleeping time was significantly shorter and the cytochrome P-450 content was significantly higher in the mice fed the FSBM-FD diet than the respective value in mice fed the other test diets . In experiment 4, mice were fed one of eight diets which contained different levels of aglycone obtained by varying the proportion of FSBM-FD and FSBM-FD-R, for 28 d . The cytochrome P-450 content in hepatic microsomes increased as the dietary level of isoflavonoid aglycones increased, but there was a saturation phenomenon . These results suggest that soy isoflavonoid aglycones are more potent inducers of cytochrome P-450 than isoflavonoid glycosides. J Agric Food Chem, 2000 Apr, 48(4), 967 - 73 Pesticide residues in grapes, wine, and their processing products; Cabras P et al.; In this review the results obtained in the 1990s from research on the behavior of pesticide residues on grapes, from treatment to harvest, and their fate in drying, wine-making, and alcoholic beverage processing are reported . The fungicide residues on grapes (cyproconazole, hexaconazole, kresoxim-methyl, myclobutanil, penconazole, tetraconazole, and triadimenol), the application rates of which were of a few tens of grams per hectare, were very low after treatment and were not detectable at harvest . Pyrimethanil residues were constant up to harvest, whereas fluazinam, cyprodinil, mepanipyrim, azoxystrobin, and fludioxonil showed different disappearance rates (t(1/2) = 4.3, 12, 12.8, 15.2, and 24 days, respectively) . The decay rate of the organophosphorus insecticides was very fast with t(1/2) ranging between 0.97 and 3.84 days . The drying process determined a fruit concentration of 4 times . Despite this, the residue levels of benalaxyl, phosalone, metalaxyl, and procymidone on sun-dried grapes equalled those on the fresh grape, whereas they were higher for iprodione (1.6 times) and lower for vinclozolin and dimethoate (one-third and one-fifth, respectively) . In the oven-drying process, benalaxyl, metalaxyl, and vinclozolin showed the same residue value in the fresh and dried fruit, whereas iprodione and procymidone resides were lower in raisins than in the fresh fruit . The wine-making process begins with the pressing of grapes . From this moment onward, because the pesticide on the grape surface comes into contact with the must, it is in a biphasic system, made up of a liquid phase (the must) and a solid phase (cake and lees), and will be apportioned between the two phases . The new fungicides have shown no effect on alcoholic or malolactic fermentation . In some cases the presence of pesticides has also stimulated the yeasts, especially Kloeckera apiculata, to produce more alcohol . After fermentation, pesticide residues in wine were always smaller than those on the grapes and in the must, except for those pesticides that did not have a preferential partition between liquid and solid phase (azoxystrobin, dimethoate, and pyrimethanil) and were present in wine at the same concentration as on the grapes . In some cases (mepanipyrim, fluazinam, and chlorpyrifos) no detectable residues were found in the wines at the end of fermentation . From a comparison of residues in wine obtained by vinification with and without skins, it can be seen that their values were generally not different . Among the clarifying substances commonly used in wine (bentonite, charcoal, gelatin, polyvinylpolypyrrolidone, potassium caseinate, and colloidal silicon dioxide), charcoal allowed the complete elimination of most pesticides, especially at low levels, whereas the other clarifying substances were ineffective . Wine and its byproducts (cake and lees) are used in the industry to produce alcohol and alcoholic beverages . Fenthion, quinalphos, and vinclozolin pass into the distillate from the lees only if present at very high concentrations, but with a very low transfer percantage (2, 1, and 0.1%, respectively) . No residue passed from the cake into the distillate, whereas fenthion and vinclozolin pass from the wine, but only at low transfer percentages (13 and 5%, respectively). Digestion, 2000, 61(3), 193 - 200 Application of the colon-simulation technique for studying the effects of Saccharomyces boulardii on basic parameters of porcine cecal microbial metabolism disturbed by clindamycin; Breves G et al.; AIMS: The present study analyzed the effects of Saccharomyces boulardii on the biochemical parameters of microbial hindgut metabolism disturbed by clindamycin . METHODS: The experiments were carried out under in vitro conditions using the semicontinuous colon-simulation technique . This technique is standardized for quantitatively measuring parameters of microbial hindgut metabolism . The fluid and particle phase of pig hindgut contents were used for the in vitro incubations . The 5-day control period was followed by clindamycin exposure alone (312.5 mg/day for 5 days) or by a combined treatment of clindamycin and S . boulardii (400 mg/day for 5 days) . RESULTS: Clindamycin resulted in significant decreases in production rates of short-chain fatty acids (SCFAs) which were associated with substantial changes in molar SCFA proportions at the expense of butyrate . These effects could at least partly be compensated for by S . boulardii, in particular by enhancements of acetate and propionate fermentation to control levels . In contrast, butyrate fermentation could not be reconstituted . In a second experiment the potential use of S . boulardii as a substrate for hindgut microbial metabolism was studied by comparing living and autoclaved yeast . Propionate and butyrate fermentation rates were unaffected whereas acetate fermentation tended to be higher in the presence of living yeast . CONCLUSIONS: S . boulardii can be effective to compensate for changes in microbial fermentation in response to antibiotic treatment . Despite the lack of statistical significance it might be concluded that the increase in fermentation end products can only partly be explained by the utilization of the yeast as a substrate for microbial metabolism . Appl Microbiol Biotechnol, 2000 Mar, 53(3), 301 - 9 Simultaneous bioconversion of glucose and xylose to ethanol by Saccharomyces cerevisiae in the presence of xylose isomerase; Chandrakant P et al.; Simultaneous isomerisation and fermentation (SIF) of xylose and simultaneous isomerisation and cofermentation (SICF) of a glucose/xylose mixture was carried out by Saccharomyces cerevisiae in the presence of xylose isomerase . The SIF of 50 g I(-1) xylose gave an ethanol concentration and metabolic yield of 7.5 g l(-1) and 0.36 g (g xylose consumed)(-1) . These parameters improved to 13.4 g l(-1) and 0.40 respectively, when borate was added to the medium . The SICF of a mixture of 50 g l(-1) glucose and 50 g l(-1) xylose gave an ethanol concentration and metabolic yield of 29.8 g l(-1) and 0.42 respectively, in the presence of borate . Temperature modulation from 30 degrees C to 35 degrees C during fermentation further enhanced the above parameters to 39 g l(-1) and 0.45 respectively . The approach was extended to the bioconversion of sugars present in a real lignocellulose hydrolysate (peanut-shell hydrolysate) to ethanol, with a fairly good yield. Appl Microbiol Biotechnol, 2000 Mar, 53(3), 282 - 8 Overexpression of the lat gene in Nocardia lactamdurans from strong heterologous promoters results in very high levels of lysine-6-aminotransferase and up to two-fold increase in cephamycin C production; Chary VK et al.; The level of lysine-6-aminotransferase (encoded by the lat gene), an enzyme that commits lysine to the cephamycin biosynthesis pathway, is very low in wild type Nocardia lactamdurans . Two lat overexpression systems (pAMEXlat and pSAFlat) were constructed to express the promoterless lat gene of N . lactamdurans from the strong promoters amyP (of the alpha-amylase gene) and safP (of the secretion activating factor gene) of Streptomyces griseus . Both constructions led to very high levels of lysine-6-aminotransferase (between 8- and 15-fold) in the cells . Expression of lat from the amy promoter was optimal in glycerol-containing medium and was negatively regulated by glucose . The high levels of lysine-6-aminotransferase resulted in a 50-200% increase in cephamycin C production in the standard fermentation conditions . Onset of cephamycin C biosynthesis occurred at the same time in control and in lat-overexpressing strains, but the cephamycin production rate was clearly higher in transformants overexpressing the lat gene . Furthermore, HPLC analysis of cephamycin C in the culture broths revealed an early depletion of biosynthetic intermediates and an accumulation of cephamycin C when the lat gene was overexpressed . These results indicate that lysine-6-aminotransferase activity is limiting for cephamycin C biosynthesis under some culture conditions. Appl Microbiol Biotechnol, 2000 Mar, 53(3), 266 - 71 Optimization of the hydroxylation of 2-cyclopentylbenzoxazole with Cunninghamella blakesleeana DSMZ 1906; Kraemer-Schafhalter A et al.; Biohydroxylation of 2-cyclopentyl-1,3-benzoxazole with the filamentous fungus Cunninghamella blakesleeana DSMZ 1906 was studied in a 15-l stirred tank reactor . The aim of the work was to avoid substrate limitation through sub-optimal mixing by formation of pellets with a uniform pellet size distribution of 250-500 microm, obtained at an inoculum concentration of 10(7) spores ml(-1) and an agitation rate of 390 rpm . Due to the high toxicity of the educt, 2-cyclopentyl-1,3-benzoxazole, on the fungus, the medium composition, the time of educt addition, and the educt starting concentration were optimized to reach high educt tolerance and hydroxylation activity . A good maintenance of biotransformation capacity was obtained without excessive loss of activity of the biocatalyst by addition of 30 mg 2-cyclopentyl-1,3-benzoxazole/g biomass (cell dry mass) during the stationary phase in a medium which was optimized in batch fermentations with experimental designs . An increase in product yield and quality (enantiomeric excess) was achieved by developing feeding strategies combining the educt and medium components . The resulting fermentation broth contained 450 mg l(-1) of the product (1S,3S)-3-(benz-1,3-oxazol-2-yl)cyclopentan-1-ol with an enantiomeric excess of 95%, which represents a 48% increase over former reported results. J Food Prot, 2000 Apr, 63(4), 441 - 4 Survival of Escherichia coli O157:H7 in buttermilk as affected by contamination point and storage temperature; McIngvale SC et al.; The effects of contamination point (during fermentation versus postfermentation) and storage temperature (5 and 12 degrees C) were determined for survival of Escherichia coli O157:H7 in fermented buttermilk . E . coli O157:H7 was recovered from buttermilk inoculated during fermentation for 22 days and in buttermilk inoculated postfermentation for 32 days . For storage temperatures of 5 and 12 degrees C, D-values were lower for E . coli O157:H7 inoculated during fermentation (2.5, 2.2 days) than postfermentation (5.6, 4.8 days) (P < 0.05) . Developed acidity in inoculated buttermilks was not different from controls (P > 0.05) . The extended recovery of viable enterohemorrhagic E . coli O157:H7 from both processing scenarios indicates that the presence of E . coli O157:H7 in buttermilk is not limited to postprocessing contamination. Enzyme Microb Technol, 2000 Apr 1, 26(7), 537 - 543 Screen-printed enzyme sensors for l-lysine determination; Olschewski H et al.; Sensors for the determination of L-lysine in samples of fermentation broth have been developed . Low-cost screen-printed sensors comprising a platinum working electrode, an Ag/AgCl pseudo reference and a carbon counter electrode were used as transducers for the enzyme sensors . L-lysine-(alpha)-oxidase from Trichoderma viride has been immobilized by entrapment into a polyurethane hydrogel . Sensors were characterized for L-lysine with respect to pH value, linear range, reproducibility, repeatability, storage and working stability . The sensitivities to other amino acids were also determined . A batch system with two working electrodes, one with immobilized enzyme and one without was adapted for the determination of L-lysine by differential measurements . Good agreement was found between L-lysine concentrations measured by the enzyme sensors and by a conventional amino acid analyzer. J Anim Sci, 2000 Mar, 78(3), 750 - 9 Ruminal fermentation and duodenal flow following progressive inoculations of fauna-free wethers with major individual species of ciliate protozoa or total fauna; Ivan M et al.; Naturally fauna-free (FF) wethers, equipped with ruminal and duodenal cannulas, were used in two groups of eight (Group A) and seven (Group B) animals in six consecutive experimental periods, each lasting for 28 d . The objective was to measure ruminal fermentation traits, and flows of nonammonia nitrogen (NAN), total amino acid (TAA), and bacterial nitrogen (BN) from the stomach after inoculation with individual ciliate protozoa species in each period . The wethers in both groups were fed a diet based on corn silage, haylage, and soybean meal, and they remained FF during the first period . At the beginning of each other period, the wethers were progressively inoculated intraruminally with one individual major species of ruminal ciliate protozoa or total fauna (TF) . Thus, Group A was progressively inoculated (+) with Dasytricha ruminantium (DS), Polyplastron multivesiculatum (PP), Isotricha intestinalis (IS), Entodinium caudatum (EN) and TF-type A . Also, Group B was progressively inoculated (+) with IS, DS, Epidinium ecaudatum (EP), Eudiplodinium maggi (EU), and EN . Duodenal digesta and ruminal fluid were collected and sampled in each period on d 26 and 28, respectively, and subjected to chemical analyses . A significantly higher (P < .05) pH (6.4) in ruminal fluid of the Group A wethers was obtained when each DS, DS+PP, DS-PP-IS+EN, and TF population was present in the rumen than when the wethers were FF (6.2) . In the Group B wethers, pH (6.1) was lower (P < .05) for the population of IS-DS-EP+EU than for other populations (6.2 to 6.3) . The concentration of total VFA in ruminal fluid was higher (P < .05) in the Group B wethers when IS, IS+DS, or IS-DS+EP populations were present in the rumen than when the wethers were FF . The flow of NAN, TAA, and BN from the stomach to the intestinal tract was generally lower for different protozoa populations than for the FF period . Largest decreases (P < .05) in the flow of NAN, TAA, and BN occurred when EN was added into the rumen of wethers in the A and B groups, which already contained populations of DS-PP+IS and IS-DS-EP+EU, respectively . Holotrich protozoa had very little effect on the protein metabolism in the rumen, but cellulolytic protozoa (PP, EP, and EU) and EN decreased the efficiency of protein utilization by the ruminant host. J Anim Sci, 2000 Mar, 78(3), 735 - 41 Pretanned leather shavings in a supplement mixture for steers: II . Digesta kinetics, ruminal fermentation, and grazing behavior in steers grazing dormant wheatgrass pasture; Mendoza-Reyes A et al.; Twelve ruminally cannulated steers (Angus x Holstein; average initial BW = 533 +/- 3.28 kg) were randomly allotted to one of three treatments (four steers/treatment) to evaluate the use of pretanned leather shavings as a component of a protein supplement for steers grazing dormant intermediate wheatgrass (Thinopyrum intermedium Host) . Steers were allotted to one of three treatments: 1) no supplement (control); 2) supplementation intraruminally at 0700 with soybean meal at .2% of BW (as-fed basis); 3) supplementation intraruminally with soybean meal and pretanned leather shavings (17:8 ratio, respectively) at .16% of BW (as-fed basis) . Supplements were formulated so that intakes were isonitrogenous and were placed intraruminally once daily (0700) . Sampling periods were conducted February 3 to 16 and February 17 to March 5, 1995 . In situ organic matter disappearance of the soybean meal supplement was greater (P > .05) than that of the leather shavings supplement at all incubation times (1, 3, 6, 9, 12, 24, and 48 h) . Data suggested that pretanned leather shavings within the leather shavings supplement were only 25% degradable within the rumen . Forage OM intake (control = 12.7, soybean meal = 12.7, and leather shavings = 13.4 g/kg of BW), grazing time, and grazing efficiency were not altered (P > .10) by supplementation or type of supplement provided but did increase between the February and March samplings . Total intake was increased (P = .09) with supplementation and reflected the addition of the protein supplements . Particulate and fluid passage estimates were unaffected (P > .10) by the supplements; however, gastrointestinal fill increased (P = .01) between the February and March samplings . Ruminal pH was lower (P = .04) and ruminal NH3 N concentration was greater (P = .02) for supplemented steers than for control steers, and supplementation treatments did not differ (P > .10) . Total VFA concentrations were increased (P = .01) by supplementation but were not affected by type of supplement provided (P > .10) . Ruminal molar proportions of acetate and propionate and the ratio of these two VFA did not differ (P > .10) between supplementation types . Nonetheless, supplementation increased molar proportions of butyrate (P = .04), valerate (P = .02), and isovalerate (P = .05), and leather shavings supplementation increased (P = .10) isobutyrate proportions over those in steers supplemented with soybean meal . Combining pretanned leather shavings with soybean meal seemed to have no deleterious effects on forage intake, digesta passage, grazing behavior, or ruminal fermentation and seemed to provide effects similar to those of soybean meal alone. J Anim Sci, 2000 Mar, 78(3), 726 - 34 Pretanned leather shavings in a supplement mixture for steers: I . In situ and in vitro disappearance, ruminal fermentation, and organic matter, nitrogen, and fiber digestion; Ackerman CJ et al.; Two digestion studies were conducted to evaluate the use of pretanned leather shavings as a component of a protein supplement . In Exp . 1, the in situ and in vitro disappearance of pretanned leather shavings and soybean meal was evaluated . Results revealed that less than 18.4% of the pretanned leather shavings was solubilized and disappeared when exposed to McDougall's buffer for 48 h, but there was 90.0% disappearance with 48-h exposure to a .1 N HCl/pepsin treatment and 97.0% disappearance with exposure to a two-stage digestion . In situ disappearance following 72 h in the rumen allowed 6.8% disappearance . Thus, leather shavings seem to be relatively indigestible in the rumen, but postruminal digestion may be possible . In Exp . 2, six Angus x Holstein steers, fitted with ruminal and duodenal cannulas, were used in a replicated 3 x 3 Latin square to evaluate ruminal and digestion effects of the following supplements combined with fescue hay at 1.7% of BW (DM basis): no supplementation (control); supplementation intraruminally with soybean meal at .07% of BW (as-fed basis); and supplementation intraruminally with a combination of soybean meal and pretanned leather shavings (17:8 ratio) at .05% of BW (isonitrogenous to soybean meal; as-fed basis) . Ruminal fluid passage rate was greater and fluid turnover time was shorter in steers fed leather shavings than in those fed soybean meal (P = .10) . Ruminal pH was lower (P = .04) for supplemented steers than for control steers and ruminal NH3 N concentration was greater (P = .01) in steers fed soybean meal than in those fed leather shavings . Total VFA concentration was increased (P = .02) by supplementation . Supplementation with soybean meal increased (P < .05) ruminal molar proportions of butyrate, valerate, and isovalerate compared with leather shavings . Duodenal OM flow and OM disappearing in the intestines were increased by supplementation (P < .10), but not by the type of supplement fed (P > .10) . Ruminal digestion of OM and total tract OM digestion were unaffected (P > .10) by supplementation and the type of supplement fed . Flow and digestion of NDF were unaffected (P > .10) by the treatments . Flow of N and the quantity of N disappearing in the intestines were increased (P < .05) by supplementation but did not differ (P > .10) between supplementation groups . Microbial N flow, N utilization for net microbial protein synthesis, and ruminal N disappearance were unaffected (P > .10) by supplementation and the type of supplement provided . Combining pretanned leather shavings with soybean meal seemed to have no deleterious effects on digestion or fermentation and to allow for escape of some N to the lower tract. J Anim Sci, 2000 Mar, 78(3), 638 - 46 Effect of fiber source on cecal fermentation and nitrogen recycled through cecotrophy in rabbits; Garcia J et al.; The influence of fiber source on fiber digestion in rabbits was investigated . Six fibrous feedstuffs with wide differences in chemical composition and particle size were selected: paprika meal, olive leaves, alfalfa hay, soybean hulls, sodium hydroxide-treated barley straw, and sunflower hulls . Six diets were formulated to contain one of these ingredients as the sole source of fiber . To avoid nutrient imbalances, fiber sources were supplemented with different proportions of a fiber-free concentrate, based on soy protein isolate, wheat flour, lard, and a vitamin and mineral mix, to obtain diets containing at least 3% nitrogen and 5% starch . Daily soft feces excretion, and its NDF, and total and microbial nitrogen content were determined in 60 fattening rabbits (10 per diet) . Seven days after the last cecotrophy control, the same animals were used to determine weight of stomach, cecum and their contents, and cecal fermentation traits (pH, VFA and ammonia concentrations, and buffer properties of cecal contents) . Stepwise regression analysis showed a positive effect (P < .001) on soft feces excretion, total and microbial nitrogen concentrations in soft feces, cecal acidity, and total VFA in the cecum of dietary pectic constituents (2.9, 3.5, 2.5, .9, and 6.6%) and proportion of fine particles (< .315 mm) (1.8, .9, 1.3, .15, and .9%) per each increment of one percentage unit of the independent variables . Proportion of fine particles also increased weight of cecal contents (P < .001) . Soft feces excretion and weight of stomach and of its contents increased (P < .001) by 5.2, 2.8, and 10.2% per each percentage unit increment of proportion of large particles (> 1.25 mm) . Degree of lignification of NDF decreased total nitrogen concentration in soft feces and cecal VFA concentration (P < .001) . Source of fiber affected cecal pH not only by its influence on the cecal concentrations of the final products of fermentation, but also through its effect on the pH of dry cecal contents (P < .001) . The latter was negatively correlated with dietary proportion of fine particles, degree of lignification of NDF, and base-buffering capacity of dry cecal contents (r = -.52, -.37, and -.49, respectively) . From these results, we conclude that pectic constituent concentration, degree of lignification of NDF, and particle size are the variables that best characterize the influence of the source of fiber on soft feces excretion and cecal fermentation traits in rabbits. Am J Gastroenterol, 2000 Apr, 95(4), 1017 - 20 Fermentation of dietary starch in humans; Ahmed R et al.; OBJECTIVE: Dietary starch that escapes digestion in the small intestine may be quantitatively more important than dietary fiber as a substrate for fermentation . The products of fermentation have important implications in the pathogenesis of colorectal cancer and other diseases of the large bowel, which are uncommon in Africans but have a high prevalence in Western populations . METHODS: Maize porridge is a staple of most blacks in South Africa . Stale maize porridge (high-resistant starch {HRS}) seems to induce greater fermentation in the large bowel than fresh maize porridge (low-resistant starch {LRS}) . RESULTS: In the present study, healthy colostomy subjects fed stale maize porridge had significantly more production of short-chain fatty acids (SCFA) (mean SCFA, HRS = 182.6; mean SCFA, LRS = 116.1; p < 0.05) in their colostomy effluent together with a significant drop in stool pH (mean pH, HRS = 5.91; mean pH, LRS = 6.70; p < 0.001) . The SCFA butyrate (mean, HRS = 35.1; mean, LRS = 17.6; p < 0.05) and acetate (mean, HRS = 93.9; mean, LRS = 65.8; p < 0.05) were significantly elevated on the stale maize porridge diet when compared with consumption of fresh maize porridge . SCFA propionate (mean, HRS = 43.1; mean, LRS = 24.8; p = 0.05), also increased with stale maize porridge, but was not statistically significant . CONCLUSION: A high-resistant starch diet and its resultant increase in fermentation products may be partly responsible for protecting the black population against colorectal cancers and other large bowel diseases. Arch Microbiol, 2000 Mar, 173(3), 206 - 12 Hydroxyhydroquinone reductase, the initial enzyme involved in the degradation of hydroxyhydroquinone (1,2,4-trihydroxybenzene) by Desulfovibrio inopinatus; Reichenbecher W et al.; The recently isolated sulfate reducer Desulfovibrio inopinatus oxidizes hydroxyhydroquinone (1,2,4trihydroxybenzene; HHQ) to 2 mol acetate and 2 mol CO2 (mol substrate)-1, with stoichiometric reduction of sulfate to sulfide . None of the key enzymes of fermentative HHQ degradation, i.e . HHQ-1,2,3,5-tetrahydroxybenzene transhydroxylase or phloroglucinol reductase, were detected in cell-free extracts of D . inopinatus, indicating that this bacterium uses a different pathway for anaerobic HHQ degradation . HHQ was reduced with NADH in cell-free extracts to a nonaromatic compound, which was identified as dihydrohydroxyhydroquinone by its retention time in HPLC separation and by HPLC-mass spectrometry . The compound was identical with the product of chemical reduction of HHQ with sodium borohydride . Dihydrohydroxyhydroquinone was converted stoichiometrically to acetate and to an unknown coproduct . HHQ reduction was an enzymatic activity which was present in the cell-free extract at 0.25-0.30 U (mg protein)-1, with a pH optimum at 7.5 . The enzyme was sensitive to sodium chloride, potassium chloride, EDTA, and o-phenanthroline, and exhibited little sensitivity towards sulfhydryl group reagents, such as copper chloride or p-chloromercuribenzoate. Am J Physiol Gastrointest Liver Physiol, 2000 Apr, 278(4), G578 - 84 Modulation by colonic fermentation of LES function in humans; Piche T et al.; Colonic fermentation of carbohydrate has been shown to influence gastric and intestinal motility . Our aim was to investigate the effects of colonic infusion of lactose and short-chain fatty acids (SCFAs) on lower esophageal sphincter (LES) function in humans . LES pressure (LESP), transient relaxations of LES (TLESRs), and esophageal pH were monitored over 6 h on 4 different days in 7 healthy volunteers . After 1 h of baseline recording, the effects of different colonic infusions (270 ml of isotonic or hypertonic saline, 30 g lactose, or 135 mmol SCFAs) were tested in fasting conditions and after a standard meal . Peptide YY (PYY) and oxyntomodulin (OLI) were also measured in plasma . Both lactose and SCFA infusions increased the number of TLESRs as well as the proportion of TLESRs associated with acid reflux episodes, but saline solutions did not . The postprandial fall of LESP was enhanced by previous SCFA infusion . Plasma PYY and OLI increased similarly after all colonic infusions . Colonic fermentation of lactose markedly affected LES function, and this effect was reproduced by SCFA infusion . Whether the mechanisms of this feedback phenomenon are of hormonal nature, neural nature, or both remains to be determined. Mol Microbiol, 2000 Mar, 35(6), 1421 - 30 Saccharomyces cerevisiae PAU genes are induced by anaerobiosis; Rachidi N et al.; Saccharomyces cerevisiae PAU genes constitute the largest multigene family in yeast, with 23 members located mainly in subtelomeric regions . The role and regulation of these genes were previously unknown . We detected PAU gene expression during alcoholic fermentation . An analysis of PAU gene regulation using PAU-lacZ fusions and Northern analyses revealed that they were regulated by anaerobiosis . PAU genes display, however, different abilities to be induced by anaerobiosis and this appears to be related to their chromosomal localization; two subtelomeric copies are more weakly inducible than an interstitial one . We show that PAU genes are negatively regulated by oxygen and repressed by haem . Examination of PAU gene expression in rox1Delta and tup1Delta strains indicates that PAU repression by oxygen is mediated by an unknown, haem-dependent pathway, which does not involve the Rox1p anaerobic repressor but requires Tup1p . Given the size of the gene family, PAU genes could be expected to be important during yeast life and some of them probably help the yeast to cope with anaerobiosis. J Am Coll Nutr, 2000 Apr, 19(2 Suppl), 100S - 110S Folates and dairy products: a critical update; Forssen KM et al.; In recent years, folates have come into focus due to their protective role against child birth defects, for example, neural tube defects . In addition, folates may have a protective role to play against coronary heart disease and certain forms of cancer . During the last few years most countries have established increased recommended intakes of folates, for example, between 300-400 microg per day for adults . This review of folates in milk and dairy products compares some recent data based on high pressure liquid chromatography (HPLC) analyses and radioprotein-binding assays, with previous data based on microbiological assays . All three methods show similar ranges for folates in cow's milk, 5-10 microg per 100 g, the variation being due to seasonal variations . Data on folates in fermented milk (buttermilk and yogurt) are also similar for these methods . Different starter cultures, however, might explain some of the variations in folate content and folate forms . Most cheese varieties contain between 10 microg and 40 microg folate per kg, with slightly higher values for whey cheese . Ripened soft cheeses may contain up to 100 microg folate per 100 g . Most previous and recent studies using HPLC indicate that 5-methyl-tetrahydrofolate (5-methyl-THF) is the major folate form in milk, but more studies are needed concerning folate forms in other, especially fermented dairy products . Relatively new data on actual concentrations in different dairy products show folate-binding proteins (FBP) to occur in unprocessed milk, but also in pasteurised milk, spray-dried skim milk powder and whey . In contrast, UHT milk, fermented milk and most cheeses only contain low levels or trace amounts. Int J Syst Evol Microbiol, 2000 Mar, 50 Pt 2, 823 - 9 Asaia bogorensis gen . nov., sp . nov., an unusual acetic acid bacterium in the alpha-Proteobacteria; Yamada Y et al.; Eight Gram-negative, aerobic, rod-shaped and peritrichously flagellated strains were isolated from flowers of the orchid tree (Bauhinia purpurea) and of plumbago (Plumbago auriculata), and from fermented glutinous rice, all collected in Indonesia . The enrichment culture approach for acetic acid bacteria was employed, involving use of sorbitol medium at pH 3.5 . All isolates grew well at pH 3.0 and 30 degrees C . They did not oxidize ethanol to acetic acid except for one strain that oxidized ethanol weakly, and 0.35% acetic acid inhibited their growth completely . However, they oxidized acetate and lactate to carbon dioxide and water . The isolates grew well on mannitol agar and on glutamate agar, and assimilated ammonium sulfate for growth on vitamin-free glucose medium . The isolates produced acid from D-glucose, D-fructose, L-sorbose, dulcitol and glycerol . The quinone system was Q-10 . DNA base composition ranged from 59.3 to 61.0 mol% G + C . Studies of DNA relatedness showed that the isolates constitute a single species . Phylogenetic analysis based on their 16S rRNA gene sequences indicated that the isolates are located in the acetic acid bacteria lineage, but distant from the genera Acetobacter, Gluconobacter, Acidomonas and Gluconacetobacter . On the basis of the above characteristics, the name Asaia bogorensis gen . nov., sp . nov . is proposed for these isolates . The type strain is isolate 71T (= NRIC 0311T = JCM 10569T). Int J Syst Evol Microbiol, 2000 Mar, 50 Pt 2, 751 - 8 Dyadobacter fermentans gen . nov., sp . nov., a novel gram-negative bacterium isolated from surface-sterilized Zea mays stems; Chelius MK et al.; A Gram-negative bacterium, designated NS114T, was isolated from duplicate treatments of surface-sterilized Zea mays stems . The plants were grown in synthetic soil under greenhouse conditions and watered with fertilizer containing no nitrogen . Strain NS114T could not be isolated from plants watered with the standard level or 20% (w/v) of the standard level of nitrogen . Cells occurred as pairs in young cultures that attached to form angled arrangements in R2A broth and occasionally formed rounded, horseshoe arrangements in YM broth . Cell variation resulted in flocculent chains of coccoid cells in old cultures . Strain NS114T fermented glucose and sucrose . The G + C content was 48 mol% . Phylogenetic analysis of the 16S rRNA gene showed that the strain was a member of the domain Bacteria and branched from a point equidistant from an aquatic organism, Runella slithyformis and a marine isolate, 'Microscilla furvescens' . Phenotypic and genotypic analyses indicated that strain NS114T could not be assigned to any recognized genus; therefore a new genus and species, Dyadobacter fermentans gen . nov., sp . nov., is proposed, for which NS114T is the type strain. Int J Syst Evol Microbiol, 2000 Mar, 50 Pt 2, 721 - 9 Orenia salinaria sp . nov., a fermentative bacterium isolated from anaerobic sediments of Mediterranean salterns; Moune S et al.; A diverse range of fermentative bacteria have been isolated from the commercial salterns of Salin-de-Giraud (Camargue, France) . One of these isolates, strain SG 3902T, has many of the morphological and physiological characteristics of the genus Orenia, as was confirmed by a phylogenetic study based on 16S rRNA gene sequencing . The closest species is Orenia marismortui, with a similarity of only 95.1% . However, strain SG 3902T, unlike O . marismortui, does not ferment mannose, glycogen or starch . The G + C contents of the DNA also differ significantly, being 29.6 mol% for O . marismortui and 33.7 mol% for strain SG 3902T . On the basis of these physiological and genetic differences, it is proposed that strain SG 3902T should be considered as a representative of a new species belonging to the genus Orenia, under the name Orenia salinaria sp . nov . The type strain is SG 3902T (= ATCC 700911). Int J Syst Evol Microbiol, 2000 Mar, 50 Pt 2, 645 - 8 Pelospora glutarica gen . nov., sp . nov., a glutarate-fermenting, strictly anaerobic, spore-forming bacterium; Matthies C et al.; The strictly anaerobic, Gram-negative, spore-forming bacterium strain WoGl3T had been enriched and isolated in mineral medium with glutarate as the sole source of energy and organic carbon . Glutarate was fermented to a mixture of butyrate, isobutyrate, CO2 and small amounts of acetate . Strain WoGl3T grew only with the dicarboxylates glutarate, methylsuccinate and succinate . 16S rDNA sequence analysis revealed an affiliation of strain WoGl3T to the family Syntrophomonadaceae . This monophyletic group is comprised of strain WoGl3T and the genera Syntrophomonas, Syntrophospora and Thermosyntropha, within the phylum of Gram-positive bacteria with a low DNA G + C content . Overall intra-group 16S rRNA sequence similarities of 89.2-93.9% document a separate phylogenetic status for strain WoGl3T . Strain WoGl3T (= DSM 6652T) is described as the type strain of a new species within a new genus, Pelospora glutarica gen . nov., sp . nov. Trends Biotechnol, 2000 May, 18(5), 197 - 202 Biotransformations monitored in situ by proton nuclear magnetic resonance spectroscopy; Brecker L et al.; One-dimensional Fourier-transform proton nuclear magnetic resonance (1H-NMR) spectroscopy can be used to study biotransformations in situ, in vivo and in aqua (1H2O) . Although an insensitive method, it rapidly provides solution-structural information of mixtures of diverse compounds that are used and formed during enzymic reactions and culture fermentations; the samples do not require any physical or chemical processing for analysis . The absolute stereochemistry of some reactions can also be determined, and assessments of metabolic fluxes made . This technique, with appropriate modifications, is of obvious value for on-line assessments of industrial fermentation processes. Nutrition, 2000 Apr, 16(4), 289 - 95 Influence of fiber fermentability on nutrient digestion in the dog; Silvio J et al.; Eight mature dogs (17.2 +/- 0.2 kg) surgically fitted with ileal T-cannulas were used in a replicated 4-x-4 Latin-square-design experiment to evaluate nutrient disappearance at the terminal ileum and through the digestive tract . Two fiber types, cellulose, a crystalline, slowly fermented fiber, and pectin, a soluble, rapidly fermented fiber, were fed in different increments, and the effects on nutrient availability were assessed . Treatments included 1) 100% cellulose, 2) 66% cellulose and 33% pectin, 3) 66% pectin and 33% cellulose, and 4) 100% pectin . Fiber was added at 10% of diet dry matter (DM) . Diets were fed at 100% of ME for maintenance and offered at 0730 and 1730 h . All periods were 21 d, which included 3 d of diet transition and 7 d of adaptation . Daily DM intake was 210 +/- 5 g . Total tract and large-intestine DM digestibility increased linearly (P < 0.01) with increased pectin . These changes in DM digestion were largely the result of changes in fiber digestion . Fermentation of total dietary fiber in the large intestine went from less than zero to 39% of ileal flow (linear, P < 0.01) . Total-tract crude-protein digestibility decreased linearly (P < 0.01) with increased pectin . This study demonstrated that fiber fermentability significantly affects digestion in the dog . Increasing fermentable fiber increased the digestion of DM and energy . However, increased fiber fermentability inversely affects crude protein digestibility . The lower crude-protein digestibility could be attributed to larger microbial protein excretion as a result of greater fermentation of pectin versus cellulose. Biochemistry, 2000 Apr 18, 39(15), 4518 - 26 A PEST-like sequence in the N-terminal cytoplasmic domain of Saccharomyces maltose permease is required for glucose-induced proteolysis and rapid inactivation of transport activity; Medintz I et al.; Maltose permease is required for maltose transport into Saccharomyces cells . Glucose addition to maltose-fermenting cells causes selective delivery of this integral plasma membrane protein to the yeast vacuole via endocytosis for degradation by resident proteases . This glucose-induced degradation is independent of the proteasome but requires ubiquitin and certain ubiquitin conjugating enzymes . We used mutation analysis to identify target sequences in Mal61/HA maltose permease involved in its selective glucose-induced degradation . A nonsense mutation was introduced at codon 581, creating a truncated functional maltose permease . Additional missense mutations were introduced into the mal61/HA-581NS allele, altering potential phosphorylation and ubiquitination sites . No significant effect was seen on the rate of glucose-induced degradation of these mutant proteins . Deletion mutations were constructed, removing residues 2-30, 31-60, 61-90, and 49-78 of the N-terminal cytoplasmic domain, as well as a missense mutation of a dileucine motif . Results indicate that the proline-, glutamate-, aspartate-, serine-, and threonine-rich (PEST) sequence found in the N-terminal cytoplasmic domain, particularly residues 49-78, is required for glucose-induced degradation of Mal61/HAp and for the rapid glucose-induced inactivation of maltose transport activity . The decreased rate of glucose-induced degradation correlates with a decrease in the level of glucose-induced ubiquitination of the DeltaPEST mutant permease . In addition, newly synthesized mutant permease proteins lacking residues 49-78 or carrying an alteration in the dileucine motif, residues 69 and 70, are resistant to glucose-induced inactivation of maltose transport activity . This N-terminal PEST-like sequence is the target of both the Rgt2p-dependent and the Glc7p-Reg1p-dependent glucose signaling pathways. Genetics, 2000 Mar, 154(3), 1013 - 23 Proteasome mutants, pre4-2 and ump1-2, suppress the essential function but not the mitochondrial RNase P function of the Saccharomyces cerevisiae gene RPM2; Lutz MS et al.; The Saccharomyces cerevisiae nuclear gene RPM2 encodes a component of the mitochondrial tRNA-processing enzyme RNase P . Cells grown on fermentable carbon sources do not require mitochondrial tRNA processing activity, but still require RPM2, indicating an additional function for the Rpm2 protein . RPM2-null cells arrest after 25 generations on fermentable media . Spontaneous mutations that suppress arrest occur with a frequency of approximately 9 x 10(-6) . The resultant mutants do not grow on nonfermentable carbon sources . We identified two loci responsible for this suppression, which encode proteins that influence proteasome function or assembly . PRE4 is an essential gene encoding the beta-7 subunit of the 20S proteasome core . A Val-to-Phe substitution within a highly conserved region of Pre4p that disrupts proteasome function suppresses the growth arrest of RPM2-null cells on fermentable media . The other locus, UMP1, encodes a chaperone involved in 20S proteasome assembly . A nonsense mutation in UMP1 also disrupts proteasome function and suppresses Deltarpm2 growth arrest . In an RPM2 wild-type background, pre4-2 and ump1-2 strains fail to grow at restrictive temperatures on nonfermentable carbon sources . These data link proteasome activity with Rpm2p and mitochondrial function. J Nat Prod, 2000 Mar, 63(3), 396 - 8 Microbial transformation of benzosampangine; Orabi KY et al.; Microbial transformation studies of the synthetic antifungal alkaloid benzosampangine (1) have revealed that 1 is metabolized by a number of microorganisms . Using a standard two-stage fermentation technique Absidia glauca (ATCC 22752), Cunninghamella blakesleeana (ATCC 8688a), Cunninghamella species (NRRL 5695), Fusarium solani f . sp . cucurbitae (CSIH #C-5), and Rhizopogon species (ATCC 36060) each produced a beta-glucopyranose conjugate of benzosampangine (2) . The identity of 2 was established on the basis of spectroscopic data. J Chromatogr A, 2000 Mar 17, 873(1), 107 - 15 Development of a headspace solid-phase microextraction procedure for the determination of free volatile fatty acids in waste waters; Abalos M et al.; An analytical procedure based on headspace solid-phase microextraction (SPME) coupled to GC-flame ionization detection/Negative Chemical Ionization Mass Spectrometry has been developed for the determination of free volatile fatty acids (C2-C7) in waste water samples . Five different coatings have been evaluated and polydimethylsiloxane-Carboxen was the only fiber that allows a successful extraction of the shortest chain fatty acids (acetic and propionic) . Several parameters such as extraction time and temperature, desorption conditions, agitation speed and sample volume have been optimized using the polydimethylsiloxane-Carboxen fiber . The linear dynamic range was over two-four orders of magnitude, depending on the acid . Procedural detection limits were in the low to medium microg/l levels and the RSDs were between 5.6% and 13.3% . To evaluate the applicability of the developed SPME procedure on real samples, fermented urban wastewaters were analysed. Biotechnol Prog, 2000 Mar-Apr, 16(2), 222 - 7 Increased heterologous protein production in Aspergillus niger fermentation through extracellular proteases inhibition by pelleted growth; Xu J et al.; The dependence of filamentous fungal protease secretion on morphology was investigated by employing the recombinant Aspergillus niger strain AB4.1{pgpdAGLAGFP} which contains a gene for the glucoamylase-GFP (green fluorescence protein) fusion protein . Different inoculum levels were used to obtain different sizes of pellet or free mycelia . The extracellular protease activity of the cultures varied with the pellet size and decreased dramatically when the morphology was changed from free mycelia to pellets . The culture with an optimal pellet size of 1.6 mm was obtained from an inoculum of 4 x 10(6) spores/mL . It resulted in a specific protease activity of 158 units/L, only one-third of that in free mycelial growth, and a maximum specific GFP yield of 0.98 mg/g (cell mass) compared to 0 . 29 mg/g for free mycelial growth with an inoculum of 10(7) spores/mL . The results indicate that this bioprocessing strategy can be effectively used to inhibit protease activity in filamentous fungal fermentation and thereby to enhance heterologous protein production. Biotechnol Prog, 2000 Mar-Apr, 16(2), 208 - 12 Effect of aeration during cell growth on ketone reactions by immobilized yeast; Gervais TR et al.; The effect of aeration during cell growth on the subsequent reduction of 2-hexanone and 2-octanone by yeast cells entrapped in calcium alginate beads was studied . The reactions were conducted using 2-propanol as a sacrificial substrate to regenerate the cofactor NAD(H), and a mixture of (S)- and (R)-alcohols was produced . The use of strictly aerobic conditions when growing the cells resulted in the highest initial reaction rates, as well as the production of only a single product (i.e., the enantiomeric excess of the (S)-alcohols was 1.0) . However, initial reaction rates decreased proportionally with fermentation time regardless of whether the yeast were grown aerobically or under both aerobic and anaerobic conditions . The data also suggest that it is the aerobic (or anaerobic) condition, rather than the cell growth phase, which is responsible for the results seen. Biotechnol Prog, 2000 Mar-Apr, 16(2), 152 - 62 Solid substrate fermentation of Monascus purpureus: growth, carbon balance, and consistency analysis; Rosenblitt A et al.; Solid substrate fermentation (SSF) of Monascus purpureus on rice is a promising new technology for obtaining natural pigments . However, before attempts can be made at maximizing pigment yield, all significant macroscopic compounds should be assayed . Here, Monascus purpureus has been grown on rice in batch mode, and the evolution of the main components, biomass, residual rice, O(2), CO(2), ethanol, acetic acid, and pigments, have been followed . This set of data, never previously studied for Monascus SSF, allowed both the performance of a macroscopic elemental balance, which accounted for 83-94% of the initial substrate carbon, and a check of data consistency . Standard consistency analysis showed a significant underestimation of the nitrogen fraction of biomass, but it was unable to discriminate the errors in the carbon balance as a result of the simultaneous presence of two gross errors in the system . A simple stoichiometric model in tandem with consistency analysis explained unaccounted carbon as an underestimation of CO(2) and ethanol . Using the simplified method to estimate ethanol, the macroscopic balance accounted for 87-99% of the initial carbon. Prikl Biokhim Mikrobiol, 2000 Jan-Feb, 36(1), 30 - 6 {Optimization of conditions for cultivation of the basidiomycete Coriolus hirsutus--producer of extracellular laccase}; Koroleva OV et al.; The effects of various factors on the biosynthesis of extracellular laccase (EC 1.14.18.1) by the basidiomycete Coriolus hirsutus (Wulf.: Fr.) Quel . no . 072 during submerged cultivation were examined . Optimal parameters for cultivation in a fermenter of 10 l were determined: temperature, 28 degrees C; stirrer rotation speed, 160 rpm; and the inoculum volume, 15% of the working volume of the fermenter . The filtrate contained peroxidase, laccase, and phenol oxidase activities and displayed a high thermal stability. Prikl Biokhim Mikrobiol, 2000 Jan-Feb, 36(1), 17 - 20 {The Use of the abundance ratio of 13C and 12C isotopes for characteristic of the origin of ethyl alcohol}; Ziakun AM et al.; During alcohol fermentation, the carbon isotope composition of ethyl alcohol produced depended on the substrate used and was characterized by the value of delta 13C equal to -24.7 +/- 0.8/1000 (wheat grain), -22 +/- 0.1/1000 (rye grain), -22 +/- 0.5/1000 (products of wood hydrolysis), -15.3 +/- 0.3/1000 (maize grain) and -10 +/- 0.1/1000 (sugar cane) . The isotope composition of carbon of ethyl alcohol obtained during catalytic hydroxylation of ethylene has a delta 13C of -30.6 +/- 0.3/1000 . The possibility of quantitative determination of specific components in mixtures of ethanol samples with various isotope compositions (chemical synthesis and alcohol fermentation of raw material from C3- or C4-plants) was shown. J Biotechnol, 2000 Mar 31, 78(3), 251 - 8 Genomics, molecular genetics and the food industry; Pridmore RD et al.; The production of foods for an increasingly informed and selective consumer requires the coordinated activities of the various branches of the food chain in order to provide convenient, wholesome, tasty, safe and affordable foods . Also, the size and complexity of the food sector ensures that no single player can control a single process from seed production, through farming and processing to a final product marketed in a retail outlet . Furthermore, the scientific advances in genome research and their exploitation via biotechnology is leading to a technology driven revolution that will have advantages for the consumer and food industry alike . The segment of food processing aids, namely industrial enzymes which have been enhanced by the use of biotechnology, has proven invaluable in the production of enzymes with greater purity and flexibility while ensuring a sustainable and cheap supply . Such enzymes produced in safe GRAS microorganisms are available today and are being used in the production of foods . A second rapidly evolving segment that is already having an impact on our foods may be found in the new genetically modified crops . While the most notorious examples today were developed by the seed companies for the agro-industry directed at the farming sector for cost saving production of the main agronomical products like soya and maize, its benefits are also being seen in the reduced use of herbicides and pesticides which will have long term benefits for the environment . Technology-driven advances for the food processing industry and the consumer are being developed and may be divided into two separate sectors that will be presented in greater detail: 1 . The application of genome research and biotechnology to the breeding and development of improved plants . This may be as an aid for the cataloging of industrially important plant varieties, the rapid identification of key quality traits for enhanced classical breeding programs, or the genetic modification of important plants for improved processing properties or health characteristics . 2 . The development of advanced microorganisms for food fermentations with improved flavor production, health or technological characteristics . Both yeasts and bacteria have been developed that fulfill these requirements, but are as yet not used in the production of foods. Curr Opin Rheumatol, 2000 Mar, 12(2), 113 - 23 Fibromyalgia, chronic fatigue syndrome, and myofascial pain syndrome; Buskila D; Fibromyalgia and widespread pain were common in Gulf War veterans with unexplained illness referred to a rheumatology clinic . Increased tenderness was demonstrated in the postmenstrual phase of the cycle compared with the intermenstrual phase in normally cycling women but not in users of oral contraceptives . Patients with fibromyalgia had high levels of symptoms that have been used to define silicone implant-associated syndrome . Tender points were found to be a common transient finding associated with acute infectious mononucleosis, but fibromyalgia was an unusual long-term outcome . The common association of fibromyalgia with other rheumatic and systemic illnesses was further explored . A preliminary study revealed a possible linkage of fibromyalgia to the HLA region . Patients with fibromyalgia were found to have an impaired ability to activate the hypothalamic pituitary portion of the hypothalamic pituitary adrenal axis as well as the sympathoadrenal system, leading to reduced corticotropin and epinephrine response to hypoglycemia . Much interest has been expressed in the literature on the possible role of autonomic dysfunction in the development or exacerbation of fatigue and other symptoms in chronic fatigue syndrome . Mycoplasma genus and mycoplasma fermentans were detected by polymerase chain reaction in patients with chronic fatigue syndrome . It was reported that myofascial temporomandibular disorder does not run in families . No major therapeutic trials in fibromyalgia, chronic fatigue syndrome, or myofascial pain syndrome were reported over the past year . The effectiveness of cognitive behavioral therapy and behavior therapy for chronic pain in adults was emphasized . A favorable outcome of fibromyalgia and chronic fatigue syndrome in children and adolescents was reported. Curr Opin Lipidol, 2000 Feb, 11(1), 49 - 56 Viscous and nonviscous fibres, nonabsorbable and low glycaemic index carbohydrates, blood lipids and coronary heart disease; Jenkins DJ et al.; Viscous fibres such as guar, glucomannans, pectins, oat betaglucan and psyllium continue to be seen as hypocholesterolaemic . Nevertheless, in large cohort studies, ironically it is the insoluble cereal fibre that has been demonstrated to relate negatively to cardiovascular disease and diabetes, despite an absence of effect on fasting lipids or postprandial glycaemia . In general, resistant or nonabsorbable starch is lipid neutral, whereas some nonabsorbable sugars or oligosaccharides may raise serum cholesterol, possibly through providing more acetate after colonic fermentation by colonic microflora . On the other hand, fructo-oligosaccharides appear to reduce serum triglycerides for reasons that are not entirely clear . Of possibly greater recent interest have been the carbohydrates that are not so much resistant to absorption, but rather are slowly absorbed . They possess some of the features of dietary fibre in providing a substrate for colonic bacterial fermentation . In the small intestine, however, they form lente or sustained release carbohydrate . In the form of low glycaemic index foods, lente carbohydrate consumption has been shown to relate to improved blood lipid profiles in hyperlipidaemic individuals and improved glycaemic control in diabetes . In larger cohort studies, low glycaemic index foods or low glycaemic load diets have been associated with higher HDL-cholesterol levels and reduced incidence of diabetes and cardiovascular disease. J Dairy Sci, 2000 Mar, 83(3), 543 - 53 Evaluation of a nonstarch polysaccharidase feed enzyme in dairy cow diets; Beauchemin KA et al.; The objective of this study was to evaluate a commercial feed enzyme product (Natugrain 33-L; BASF Corporation, Ludwigshafen, Germany) used mainly in poultry diets for use in ruminant diets . The product contained mainly beta-glucanase, xylanase, and endocellulase activities . The study was conducted as a double 3 x 3 Latin square design with six lactating dairy cows (84 +/- 12 DIM) to measure intake, chewing activities, total tract digestion, and milk production . An additional three cows fitted with ruminal cannulae were used to measure dietary effects on ruminal fermentation and in sacco digestion characteristics . Cows received a diet consisting of 45% forage containing 0, 1.22, or 3.67 L of enzyme product/tonne of total mixed ration (DM basis) . Using a low or high concentration of enzyme supplementation increased feed intake, but total tract digestibility only increased with the low concentration of enzyme . As a result, intake of digestible nutrients was increased to a greater extent for cows fed the low concentration than for cows fed the high concentration . Because the cows used in this experiment were in positive energy balance, increased intake of digestible energy due to enzyme supplementation did not increase milk yield or milk component yield . Further research is necessary to determine the mechanism by which adding a fibrolytic enzyme mixture enhanced intake, but only increased feed digestion when used at a low level. J Dairy Sci, 2000 Mar, 83(3), 518 - 25 Metabolic alterations associated with an attempt to induce laminitis in dairy calves; Momcilovic D et al.; The objective of this study was to investigate metabolic alterations in young ruminating calves associated with the sudden introduction of readily fermentable diets in an attempt to induce laminitis . Sixteen dairy bull calves, at 17 wk of age were fed equal amounts of one of four diets that contained either 71 or 81% total digestible nutrients (TDN) and 15 or 20% crude protein in a 2 x 2 factorial with time as a factor . Jugular blood and ruminal fluid were sampled, and hoof temperature was measured postfeeding at frequent intervals over a subsequent 2-d period . Hooves were examined for abnormalities and the orientation of the pedal bone radiographed prior to the experiment, 48 to 72 h into the experiment and at 3 and 7 mo later . Calves responded acutely to the 81% TDN diets by inappetence, stiffness, and diarrhea . Ruminal pH was lower and both D- and L-lactate concentrations were greater in the rumens of calves fed the 81% TDN diets . Total ruminal volatile fatty acid concentration decreased as pH declined . Whole blood L-lactate did not differ across treatments, but blood D-lactate increased in calves fed the 81% TDN diets, peaking at 32 h (7.2 mM) . Hoof temperature responses could not be explained by dietary treatments . Laminitis was not detected despite the reduction of ruminal pH and a manyfold increase in blood D-lactate. J Appl Physiol, 2000 Apr, 88(4), 1467 - 73 Historical perspectives of cellular oxygen sensing and responses to hypoxia; Lahiri S; The responses to acute and chronic hypoxia begin with oxygen sensing, and this historical perspective is written in line with this concept . The earliest pertinent work started with studies on fermentation in yeast in the 17th century, before the discovery of oxygen . It required 200 yr to localize the oxygen sensing within the cells and another 100 yr to discover the cellular oxidation reactions . Today, the consensus is that the mitochondrial respiratory chain is in part the site of oxygen sensing . In addition, membrane-bound NAD(P)H oxidase possibly takes part in oxygen sensing . Oxygen-sensing mechanisms occur in a tissue-specific fashion . For example, the carotid body responds to hypoxia promptly by eliciting a ventilatory response, whereas erythropoietin production in response to hypoxia requires more time, involving new expression of genes . The mechanism has therefore moved from the cells to genes. J Appl Microbiol, 2000 Mar, 88(3), 371 - 8 Re-assessment of the influence of yeast strain and environmental factors on glycerol production in wine; Remize F et al.; Increasing glycerol production is of concern for wine-makers in improving the quality of certain wines . We have compared the impact of strain and relevant environmental factors influencing glycerol production under the same conditions, i.e . standardized conditions simulating enological fermentation . The glycerol production of 19 industrial wine strains ranged from 6.4 to 8.9 g l-1 and varied significantly between strains . The production of acetate and succinate was also found to differ substantially depending on the strain but no significant strain-dependent variation was observed for acetaldehyde . Interestingly, high glycerol production was not correlated to high production of acetate or acetaldehyde, which are undesirable in wine . A detailed study with two low or two high glycerol-producing strains showed that temperature and the initial concentration of nitrogen had little effect on the amount of glycerol formed, although agitation or a nitrogen source composed mainly of ammoniacal nitrogen slightly enhanced glycerol production . The influence of environmental factors remained minor while the predominant factor for glycerol variability in wine was attributed to the strain . Taking into account wine-making constraints, the results indicate that achieving a high glycerol content in wine requires the selection or improvement of yeast strains rather than the control of growth and cultivation conditions. Phytochemistry, 2000 Mar, 53(6), 675 - 8 Microbial transformation of papaveraldine; El Sayed KA; Preparative-scale fermentation of papaveraldine (1), the known benzylisoquinoline alkaloid, with Mucor ramannianus 1839 (sih) has resulted in a stereoselective reduction of the ketone group and the isolation of S-papaverinol (2) and S-papaverinol N-oxide (3) . The structure elucidation of both metabolites was based primarily on 1D-, 2D-NMR analyses and chemical transformations . The absolute configuration of 2 was determined using Horeau's method of asymmetric esterification . These metabolism results were consistent with the previous plant cell transformation studies on papaverine and isopapaverine. Biotechnol Bioeng, 2000 May 5, 68(3), 345 - 52 Bacterial cellulose production by Acetobacter xylinum in a 50-L internal-loop airlift reactor; Chao Y et al.; Bacterial cellulose (BC) production was realized in a batch cultivation of Acetobacter xylinum subsp . sucrofermentans BPR2001 in a 50-L internal-loop airlift reactor . When the bacterium was cultivated with air supply, 3.8 g/L of BC was produced after 67 hours . When oxygen-enriched gas was supplied, the concentration of BC was doubled and the production rate of BC was 0.116 g/L . h, which was two times higher than that of air-supplied culture and comparable to that in a mechanically agitated stirred-tank fermentor . Bacterial cellulose produced by the airlift reactor formed a unique ellipse pellet (BC pellet), different from the fibrous form which was produced in an agitated stirred-tank fermentor . The BC-pellet suspension was demonstrated to have a higher volumetric oxygen transfer coefficient than the fibrous BC suspension in a 50-L internal-loop airlift reactor . The mixing time of BC-pellet suspension in the airlift reactor was also shorter than that in water . Biotechnol Bioeng, 2000 May 5, 68(3), 285 - 97 Dynamic biochemical reaction process analysis and pathway modification predictions; Conejeros R et al.; Recently, the area of model predictive modification of biochemical pathways has received attention with the aim to increase the productivity of microbial systems . In this study, we present a generalization of previous work, where, using a sensitivity study over the fermentation as a dynamic system, the optimal selection of reaction steps for modification (amplification or attenuation) is determined . The influence of metabolites in the activity of enzymes has also been considered (through activation or inhibition) . We further introduce a new concept in the dynamic modeling of biochemical reaction systems including a generalized continuous superstructure in which two artificial multiplicative terms are included to account for: (a) enzyme overexpression or underexpression (attenuation or amplification) for the whole enzyme pool; and (b) modification of the apparent order of a kinetic expression with respect to the concentration of a metabolite or any subset of metabolites participating in the pathway . This new formulation allows the prediction of the sensitivity of the pathway performance index (objective function) with respect to the concentration of the enzyme, as well as the interaction of the enzyme with other metabolites . Using this framework, a case study for the production of penicillin V is analyzed, obtaining the most sensitive reaction steps (or bottlenecks) and the most significant regulations of the system, due to the effect of concentration of intracellular metabolites on the activity of each enzyme . Biotechnol Bioeng, 2000 May 5, 68(3), 252 - 9 Physiological studies in aerobic batch cultivations of Saccharomyces cerevisiae strains harboring the MEL1 gene; Ostergaard S et al.; Physiological studies of Saccharomyces cerevisiae strains harboring the MEL1 gene were carried out in aerobic batch cultivations on glucose-galactose mixtures and on the disaccharide melibiose, which is hydrolyzed by the enzyme melibiase (Mel1, EC 3.2.1.22) into a glucose and a galactose moiety . The strains examined (T200, T256, M24, and TH1) were all derived from the bakers' and distillers' strain of S . cerevisiae, DGI 342 . All the strains showed a significant higher ethanol yield when growing on glucose, and half the biomass yield, compared with growth on galactose . The maximum specific uptake rates were 2.5-3.3-fold higher on glucose than on galactose for all the strains examined, and hence, ethanol production was pronounced on glucose due to respiro-fermentative metabolism . The T256 strain and the T200 strain having the MEL1 gene inserted in the HXK2 locus and the LEU2 locus, respectively, hydrolyzed melibiose with low specific hydrolysis rates of 0.03 C-mol/g/h and 0.04 C-mol/g/h, respectively . This resulted in high biomass yields on melibiose in the order of 10 g/C-mol compared with 3.7 g/C-mol for M24 and 1.6 g/C-mol for TH1 . The M24 strain, constructed by classical breeding, and the mig1/gal80 disrupted and melibiase-producing strain TH1, were superior in their ability to hydrolyze melibiose into glucose and galactose showing specific melibiose hydrolysis rates of 0.17 C-mol/g/h and 0.24 C-mol/g/h, respectively . Hence, high ethanol yields on melibiose were obtained with these two strains . Growth on the glucose-galactose mixtures showed a reduction of glucose control successfully obtained in the M24 strain and the TH1 strain . J Biol Chem, 2000 Apr 7, 275(14), 10492 - 7 ASC1/RAS2 suppresses the growth defect on glycerol caused by the atp1-2 mutation in the yeast Saccharomyces cerevisiae; Mabuchi T et al.; To better define the regulatory role of the F(1)-ATPase alpha-subunit in the catalytic cycle of the ATP synthase complex, we isolated suppressors of mutations occurring in ATP1, the gene for the alpha-subunit in Saccharomyces cerevisiae . First, two atp1 mutations (atp1-1 and atp1-2) were characterized that prevent the growth of yeast on non-fermentable carbon sources . Both mutants contained full-length F(1)alpha-subunit proteins in mitochondria, but in lower amounts than that in the parental strain . Both mutants exhibited barely measurable F(1)-ATPase activity . The primary mutations in atp1-1 and atp1-2 were identified as Thr(383) --> Ile and Gly(291) --> Asp, respectively . From recent structural data, position 383 lies within the catalytic site . Position 291 is located near the region affecting subunit-subunit interaction with the F(1)beta-subunit . An unlinked suppressor gene, ASC1 (alpha-subunit complementing) of the atp1-2 mutation (Gly(291) --> Asp) restored the growth defect phenotype on glycerol, but did not suppress either atp1-1 or the deletion mutant Deltaatp1 . Sequence analysis revealed that ASC1 was allelic with RAS2, a G-protein growth regulator . The introduction of ASC1/RAS2 into the atp1-2 mutant increased the F(1)-ATPase enzyme activity in this mutant when the transformant was grown on glycerol . The possible mechanisms of ASC1/RAS2 suppression of atp1-2 are discussed; we suggest that RAS2 is part of the regulatory circuit involved in the control of F(1)-ATPase subunit levels in mitochondria. Br J Nutr, 2000 Feb, 83(2), 143 - 50 Estimating the extent of degradation of ruminant feeds from a description of their gas production profiles observed in vitro: derivation of models and other mathematical considerations; France J et al.; Equations to describe gas production profiles, obtained using manual or automated systems for in vitro fermentation of ruminant feeds, were derived from first principles by considering a simple three-pool scheme . The pools represented were the potentially degradable and undegradable feed fractions, and accumulated gases . The equations derived and investigated mathematically were the generalized Mitscherlich, generalized Michaelis-Menten, Gompertz, and logistic . They were obtained by allowing the fractional rate of degradation to vary with time . The equations permit the extent of ruminal degradation (hence the supply of microbial protein to the duodenum) to be evaluated, thus linking the gas production technique to animal production. Br J Nutr, 1999 Aug, 82(2), 149 - 57 Effect of supplementing a fibre basal diet with different nitrogen forms on ruminal fermentation and microbial growth in an in vitro semi-continuous culture system (RUSITEC) Carro MD, Miller EL. Incubation trials were carried out with the rumen simulation technique (RUSITEC) to study the effects of four forms of N on the growth of ruminal micro-organisms and the fermentation variables when an all-fibre basal diet was incubated . The basal diet consisted of 10 g neutral-detergent fibre (NDF) from grass hay plus 2 g NDF from sugarbeet pulp . N forms were isolated soyabean protein, soyabean peptides, amino acids blended to profile soyabean protein and NH3 as NH4Cl . Half of the daily N supply was infused as NH4Cl and the other half was infused as each of the four treatments described . Non-NH3 N (NAN) forms increased NDF (P = 0.006), acid-detergent fibre (P = 0.003) and cellulose (P = 0.015) disappearance after 48 h incubation, CO2 (P < 0.001), CH4 (P = 0.002) and total volatile fatty acids production (P < 0.001), as well as the molar percentages of isobutyrate, isovalerate and valerate, which reflected the fermentation of amino acid C skeletons . NAN treatments also increased microbial N flow (P < 0.001) compared with NH3, with peptides and protein supporting more (P = 0.036) than amino acids . The proportion of microbial N derived from NH3 decreased successively (P < 0.05) with NH3 > amino acids > peptides > protein treatments, indicating preferential uptake of peptides without passage through the NH3 pool . Microbial efficiency (g microbial N/kg organic matter apparent disappearance) was greater (P = 0.002) for the NAN forms than for the NH3 treatment, with peptides and protein treatments supporting higher (P = 0.009) values than amino acid treatment . These results indicate that N forms other than NH3 are required for optimal fibre digestion and microbial growth. Appl Environ Microbiol, 2000 Apr, 66(4), 1435 - 43 Efficient improvement of silage additives by using genetic algorithms; Davies ZS et al.; The enormous variety of substances which may be added to forage in order to manipulate and improve the ensilage process presents an empirical, combinatorial optimization problem of great complexity . To investigate the utility of genetic algorithms for designing effective silage additive combinations, a series of small-scale proof of principle silage experiments were performed with fresh ryegrass . Having established that significant biochemical changes occur over an ensilage period as short as 2 days, we performed a series of experiments in which we used 50 silage additive combinations (prepared by using eight bacterial and other additives, each of which was added at six different levels, including zero {i.e . , no additive}) . The decrease in pH, the increase in lactate concentration, and the free amino acid concentration were measured after 2 days and used to calculate a "fitness" value that indicated the quality of the silage (compared to a control silage made without additives) . This analysis also included a "cost" element to account for different total additive levels . In the initial experiment additive levels were selected randomly, but subsequently a genetic algorithm program was used to suggest new additive combinations based on the fitness values determined in the preceding experiments . The result was very efficient selection for silages in which large decreases in pH and high levels of lactate occurred along with low levels of free amino acids . During the series of five experiments, each of which comprised 50 treatments, there was a steady increase in the amount of lactate that accumulated; the best treatment combination was that used in the last experiment, which produced 4.6 times more lactate than the untreated silage . The additive combinations that were found to yield the highest fitness values in the final (fifth) experiment were assessed to determine a range of biochemical and microbiological quality parameters during full-term silage fermentation . We found that these combinations compared favorably both with uninoculated silage and with a commercial silage additive . The evolutionary computing methods described here are a convenient and efficient approach for designing silage additives. Trends Biotechnol, 2000 Apr, 18(4), 136 - 41 Issues in the development of an industrial bioprocess advisory system; Glassey J et al.; The background and motivation for the construction of a fault detection and advisory system for an industrial fermentation process plant are described . Here, the knowledge extracted from the operators (implemented in the form of production rules) is integrated with multivariate data-based methods for fault detection . The industrial benefits arising from this integrated system include: (1) reduced variability, (2) increased mean performance levels, (3) reduced operator-training time and (4) knowledge management in the broader organization. J Microbiol Methods, 2000 Mar, 40(1), 99 - 104 A rapid technique for screening of lovastatin-producing strains of Aspergillus terreus by agar plug and Neurospora crassa bioassay; Kumar MS et al.; The success of strain improvement programme depends on the number of isolates that can be screened after mutagenic treatment . A technique to rapidly screen large number of high-yielding isolates was developed . The 'agar plug' method that utilizes the anti-fungal property of lovastatin to produce a zone of inhibition against Neurospora crassa was not only economical but also less labour-intensive . We were able to isolate a high-yielding strain, the productivity of which increased by 138% as compared to the parent strain in the submerged fermentation process. Z Naturforsch {C}, 2000 Jan-Feb, 55(1-2), 10 - 5 Downstream processing for xylitol recovery from fermented sugar cane bagasse hydrolysate using aluminium polychloride; Silva SS et al.; Xylitol, a sweetener comparable to sucrose, is anticariogenic and can be consumed by diabetics . This sugar has been employed successfully in many foods and pharmaceutical products . The discovery of microorganisms capable of converting xylose present in lignocellulosic biomass into xylitol offers the opportunity of producing this poliol in a simple way . Xylitol production by biotechnological means using sugar cane bagasse is under study in our laboratories, and fermentation parameters have already been established . However, the downstream processing for xylitol recovery is still a bottleneck on which there is only a few data available in the literature . The present study deals with xylitol recovery from fermented sugar cane bagasse hydrolysate using 5.2 g/l of aluminium polychloride associated with activated charcoal . The experiments were performed at pH 9, 50 degrees C for 50 min . The results showed that aluminium polychloride and activated charcoal promoted a 93.5% reduction in phenolic compounds and a 9.7% loss of xylitol from the fermented medium, which became more discoloured, facilitating the xylitol separation. J Gene Med, 1999 Sep-Oct, 1(5), 312 - 21 Insertion of natural intron 6a-6b into a human cDNA-derived gene therapy vector for cystic fibrosis improves plasmid stability and permits facile RNA/DNA discrimination; Boyd AC et al.; BACKGROUND: The gene therapy vector pCMV-CFTR containing human CFTR cDNA shows high segregational instability during growth in Escherichia coli . METHODS: By host strain screening and optimization of fermentation, satisfactory levels of pCMV-CFTR production were achieved . However, the vector was also vulnerable to structural instability manifested by the appearance during fermentation of a more stable mutant form in which the bacterial insertion sequence IS1 had transposed into exon 7 of plasmidborne CFTR . The instability of pCMV-CFTR is attributable to transcription from an upstream cryptic promoter leading to the production of CFTR peptide fragments known to be toxic when expressed in E . coli . To address this, we inserted the 1.1 kb natural human 6a-6b intron into pCMV-CFTR . RESULTS: The new vector pCMV-CFTR-int6ab is more stable in E . coli than either pCMV-CFTR or the IS1 mutant, grows to high cell density giving higher DNA yields and expresses CFTR appropriately in transfected cells . Thus, the intron has a stabilizing effect comparable to the IS1 insertion yet retains full functionality for gene therapy . We describe a PCR assay using primers directed to sequences flanking the intron that allows differentiation between DNA and mature mRNA . The T936C mutation present only in vector DNA has also been exploited to allow transgene CFTR to be distinguished and its dose-dependent expression to be detected in human cellular backgrounds . CONCLUSIONS: Instability of a plasmid vector for gene therapy has been minimized by rational modification . The introduction of an intron for this purpose offers the additional advantage of providing a discriminatory RT-PCR assay. Biosci Biotechnol Biochem, 2000 Feb, 64(2), 229 - 36 Cloning and characterization of a gene complementing the mutation of an ethanol-sensitive mutant of sake yeast; Inoue T et al.; Ethanol-sensitive mutants (esl to es10) were isolated from sake yeast, Saccharomyces cerevisiae SY-32 . These mutants were unable to grow at 7% ethanol at which the wild type strain SY-32 does grow . The mutants had a variety of fermentation rates and viabilities in the presence of ethanol . The gene ERG6, complementing the ethanol-sensitive mutation of es5, was cloned from an SY-32 gene library . ERG6 encodes S-adenosylmethionine: delta 24-sterol-C-methyltransferase (EC 2.1.1.41) in the ergosterol synthetic pathway . Mutant es5 had a reduced ability to synthesize ergosterol . An erg6 disruptant was also ethanol-sensitive . These results suggested that ERG6 plays an important role in the ethanol tolerance of S . cerevisiae. J Appl Microbiol, 2000 Feb, 88(2), 191 - 201 Growth kinetics of Acetobacterium sp . on methanol-formate in continuous culture; Bainotti AE et al.; The fermentative metabolism of Acetobacterium sp . grown on methanol-formate in continuous culture is described . The reaction stoichiometry of methanol-formate, including cells, were as follows: CH3OH + 1.13HCOOH --> 0.87CH3COOH + 0.47 cell C . Formate enhanced growth yields by approximately 60% compared with methanol-CO2-grown cultures . Comparison of yields on methanol-formate allowed calculation of an energy yield of 1.3 mol ATP per mol acetate formed during homoacetate fermentation . The magnitudes of YEG,the theoretical maximum yield of YE, and m, the maintenance coefficient, were determined by growing the organism in methanol-formate and resulted in 16.5 g cell (mol methanol catabolized)-1 and 0.674 mmol methanol catabolized (g cell)-1 h-1, respectively . It is concluded that formate might replace CO2 as a source of carboxyl donor. Bioseparation, 1999, 8(1-5), 237 - 45 Binding assays in heterogeneous media using a flow injection system with an expanded micro-bed adsorption column; Mattiasson B et al.; Competitive binding assays have been performed in flow injection systems . To further increase the versatility of the system, and to enable it to deal with samples containing particulate matter, the adsorption step was designed as an expanded bed column . Immunochemical quantification of human serum albumin was chosen as a model system to use for the development of the technology . A competitive ELISA was set up using peroxidase labelled HSA as competing ligand . The introduction of the expanded bed immunosorption column made the system tolerant to samples containing suspended particulate matter . The analytical outcome is very similar to that from the packed bed system even though more time is required for each assay cycle . The capability of the system was tested by addition of increasing amounts of yeast cells . The results clearly indicate that the system is suitable e.g . for process monitoring of fermentations. Bioseparation, 1999, 8(1-5), 153 - 8 A simplified purification procedure of alpha-lactalbumin from milk using Ca(2+)-dependent adsorption in hydrophobic expanded bed chromatography; Noppe W et al.; The technique of expanded bed adsorption is originally designed for a direct recovery of proteins from fermentor feedstocks . In this article we describe the use of expanded bed adsorption for the recovery of alpha-lactalbumins from defatted milk using the hydrophobic Streamline Phenyl gel . alpha-Lactalbumins are Ca(2+)-binding proteins . Upon Ca2+ removal, they undergo a significant conformation change rendering them more hydrophobic . Based on this unique property we develop a protocol for fast and efficient purification of alpha-lactalbumin from milk . The use of this technique results in a reduction of the number of chromatographic purification steps. Zh Obshch Biol, 2000 Jan-Feb, 61(1), 64 - 73 {Light-dependent changes in the enzyme activity of the ascorbate-glutathione cycle and ascorbate oxidase in the leaves of pea}; Mittova VO et al.; Light-determined activation of ferments of ascorbate-glutation cycle, ascorbate-oxidase in chloroplasts and cytosol is demonstrated as well as ascorbate-peroxidase, monodehydroascorbate-reductase, glutation-reductase and ascorbate-oxydase in mitochondria . On the other hands activity of mitochondrial dehydroascorbate-reductase increased on reduction of light most likely due to function of electron transport from glutation to dehydroascorbate in mitochondria . Glutation metabolism is proved to be endogenic catalytic process where the amount reconstructed glutation changes slowly with a delay and gradually follow light changes . Light dependable changes of glutation content in chloroplasts ensure resistance of ferment system again hydrogen peroxide and superoxide radicals that generate intensively at light. FEMS Microbiol Lett, 2000 Apr 1, 185(1), 51 - 7 Novel growth characteristics and high rates of nitrate reduction of an Escherichia coli strain, LCB2048, that expresses only a periplasmic nitrate reductase; Potter LC et al.; Escherichia coli strain LCB2048 is a double mutant defective in the synthesis of the two membrane-associated nitrate reductases A and Z . This strain can grow anaerobically on a non-fermentable carbon source, glycerol, in the presence of nitrate even in media supplemented with high concentrations of tungstate . This growth was totally dependent upon a highly active, periplasmic nitrate reductase (Nap) . Due to the presence of a previously unreported narL mutation, synthesis of the periplasmic nitrate reductase by this strain was induced during anaerobic growth by nitrate . We have also demonstrated that methyl viologen is an ineffective electron donor to Nap: its use leads to an underestimation of the contribution of Nap activity to the rate of nitrate reduction in vivo. Appl Occup Environ Hyg, 1999 Jan, 14(1), 34 - 8 Silo gas exposure in New York state following the dry growing season of 1995; Pavelchak N et al.; Exposure to silo gas is a recognized agricultural hazard . Silo gas produced from corn fermentation may consist of oxides of nitrogen and carbon dioxide . The presence of potentially lethal concentrations of nitrogen dioxide (NO2) within vertical silos has been well documented . The risk of silo gas exposure from other silage storage methodologies--including horizontal "ag-bags" and concrete bunkers--has been less well characterized . A dry growing season is known to be a factor for elevating nitrate levels in corn plants and can result in increased NO2 production . Farms in the northeastern United States faced drought conditions during the 1995 growing season . The New York State (NYS) Department of Health (DOH) and the New York Center for Agricultural Medicine and Health (NYCAMH) investigated four exposure incidents involving six farmworkers during September/October 1995 . Four of these workers were hospitalized for multiple days, with two workers receiving treatment in intensive care units . The remaining two workers were treated in hospital emergency departments; one refused admission and left against medical advice . We monitored NO2 levels from "ag-bags" at several New York farms . For four days, outdoor concentrations of NO2 at one site remained in excess of the National Institute for Occupational Safety and Health's (NIOSH) immediately dangerous to life and health value (IDLH) of 20 ppm . As a result of the clinical and industrial hygiene data, and the growing season's abnormal weather conditions, DOH and NYCAMH issued statewide health hazard alerts and conducted educational activities to warn farmers and their families . The findings of this study reinforce the potential hazards associated with silo gas exposure and identify the use of ag-bags as a relatively new avenue for significant worker exposure. Theriogenology, 1999 May, 51(7), 1259 - 66 Effect of type and quantity of concentrates on superovulation and embryo yield in beef heifers; Yaakub H et al.; Embryo yield and quality can be decreased following superovulation of cattle on high levels of concentrates . Concentrate type can alter rumen fermentation patterns and so affect energy availability and thus embryo quality . The objectives of this experiment were to examine the effect of 2 levels and 2 types of concentrate on superovulatory response and embryo quality in beef heifers . Beef heifers received grass silage as a basal diet and barley at 3 kg (n = 20) or ad-libitum (n = 19), or citrus/beet pulp at 3 kg (n = 18) or ad-libitum (n = 19) as the source of concentrate supplement . Silage was available ad-libitum for heifers offered 3 kg but was restricted to 1 kg DM/day for heifers on ad-libitum concentrate intake . Both concentrates contained 14% crude protein . After 100 d, heifers were treated with an intravaginal progesterone releasing device (CIDR) for 7 d, and superovulation was initiated 60 h before CIDR withdrawal . Heifers received pFSH (a total of 265 mg NIH-FSH-P1 equivalent) administered over 8 injections at 12 h intervals with the last 2 injections administered at 12 and 24 h after CIDR withdrawal; they were inseminated at 56 and 72 h after CIDR withdrawal without reference to estrus . Heifers were slaughtered 6, 7 or 8 d after the first insemination, and embryos were flushed from the uterus with PBS and were graded morphologically . Statistical analyses were performed using analysis of variance . There was no interaction between level and type of diet, and thus data for the main effects are presented separately . Final live weights at the end of the experiment and carcass weights for heifers offered ad-libitum concentrate (643.8+/-6.9 kg; 354+/-8 kg, respectively) were higher (P<0.01) than those of heifers offered 3 kg concentrate (613.1+/-7.5 kg; 338.4+/-3.2 kg, respectively) while live weights and carcass weights of heifers offered barley (625.6+/-8.1 and 345.4+/-4.1 kg, respectively) or citrus/beet pulp concentrate (631.4+/-7.0 and 348.0+/-3.1 kg, respectively) were not different (P>0.05) . Superovulatory responses (number of corpora lutea at slaughter) was greater (P<0.06) when heifers were offered 3 kg (15.5+/-1.6) than when offered ad-libitum concentrates (12.3+/-1.4) . However, the superovulatory response for both citrus/beet pulp (14.4+/-1.5) and barley (13.4+/-1.5) diets were not different (P>0.05) . Heifers offered 3 kg concentrates produced greater (P<0.05) numbers of transferable embryos (4.8+/-0.7) compared with heifers fed ad-libitum concentrates (2.8+/-0.4) . Heifers offered citrus/beet pulp produced greater (P<0.05) numbers of transferable embryos (4.8+/-0.7) than heifers offered barley (2.9+/-0.5) . These data indicate that high concentrate intake has a negative effect on embryo yield and quality and that a barley compared with citrus/beet pulp based concentrate has a negative effect on embryo quality following superovulation in beef heifers. J Agric Food Chem, 2000 Mar, 48(3), 895 - 900 Solid-state bioconversion of phenolics from cranberry pomace and role of Lentinus edodes beta-glucosidase; Zheng Z et al.; Cranberry pomace contains large amounts of phenolic glycosides, which are important sources of free phenolics that have many food uses such as antioxidants, flavorings, and nutraceuticals . Our hypothesis was that these glycosides in cranberry pomace could be hydrolyzed by beta-glucosidase produced by Lentinus edodes during solid-state fermentation . On the basis of this hypothesis, our objective was to investigate the potential of using cranberry pomace as a substrate for the production of free phenolics and beta-glucosidase through solid-state fermentation by a food-grade fungus L . edodes . Our results suggested that L . edodes beta-glucosidase played a major role in release of phenolic aglycons from cranberry pomace during solid-state fermentation . After 50 days of cultivation, the yield of total free phenolics reached the maximum of 0.5 mg per g of pomace, while the beta-glucosidase activity was about 9 units per g of pomace . The enzyme exhibited optimal activity at 60 degrees C and at pH 3.5 and was stable at temperatures up to 50 degrees C and between pH 3 and 6.5 . The major free phenolics produced from cranberry pomace were identified by HPLC as gallic acid, chlorogenic acid, p-hydroxybenzoic acid, and p-coumaric acid . These results suggest that cranberry pomace is a potential substrate for producing food-grade phenolics and fungal beta-glucosidase . The L . edodes beta-glucosidase showed good stability and tolerance to low pH and, therefore has potential applications in wine and juice processing for aroma and flavor enrichment through enzymatic hydrolysis of glucoside precursors. J Agric Food Chem, 2000 Mar, 48(3), 885 - 9 Fermentation of white wines in the presence of wood chips of American and French oak; Perez-Coello MS et al.; Must obtained from Airen grapes was fermented in the presence of wood chips (4 and 7 g/L) of either French oak (from Vosges, central France, and Allier zones) or American oak . Fermentation yields were higher than in the control fermentations carried out in the absence of wood, and production of volatile substances during fermentation (alcohols, esters, and acetates) was also higher . The volatile substances that leached out of the wood were analyzed by GC-MS-SIR . The results showed that their concentrations depended on the type and amount of the oak; some of these substances were consumed in part by the yeasts during fermentation . A taste panel favorably assessed the wines produced by fermentation in the presence of oak chips, which retained part of the must original fruity aroma. Adv Exp Med Biol, 1999, 467, 671 - 7 Determination of tryptophan and tryptophan metabolites in grape must and wine; Hoenicke K et al.; Tryptophan (Trp) and its metabolites, especially indole-3-acetic acid (IAA), are considered as potential precursors of 2-aminoacetophenone (AAP), an aroma compound which causes the "untypical aging off-flavor" (UTA) in Vitis vinifera white wines . In this study RP-HPLC with fluorescence detection was used for the qualitative and quantitative analysis of Trp and Trp-metabolites in 39 grapes, 22 grape musts and 16 wines, to which different viticultural conditions (ripeness, pruning, strip of leaves, soil condition) have been applied . A sensitive and selective determination was achieved after solid phase extraction using an anion exchange material . Only traces of Trp-metabolites could be determined in the examined grapes and grape musts, but their amounts increased significantly during fermentation, whereas the amount of Trp decreased . Different viticultural measures, besides the time of grape harvest, showed no significant influences on the amount of Trp and Trp-metabolites. Adv Exp Med Biol, 1999, 467, 469 - 80 Synthesis, formation, and occurrence of contaminants in biotechnologically manufactured L-tryptophan; Simat TJ et al.; The pattern of contaminants in pharmaceutical and feed grade L-tryptophan (Trp) was investigated in a market survey of 22 lots of 6 different manufacturers . To date, 5 case associated contaminants in Showa Denko tryptophan (SD-Trp) known to cause the autoimmune disease eosinophilia-myalgia syndrome (EMS) have been structurally elucidated: 3a-hydroxy-1,2,3,3a,8,8a-hexahydropyrroloindole-2-carboxylic acid (PIC), an indoline compound, is one of the most abundant degradation compounds of unbound Trp during oxidative treatment . 2-(3-indolylmethyl)-L-tryptophan (IMT) and 2-(2-hydroxyindoline)-tryptophan (HIT) are both 2-substituted Trp-derivatives . IMT was synthesized by the reaction of Trp and indole-3-methanol or indole-3-acetaldehyde, respectively . From this finding it is proposed that Trp-metabolites can decompose under formation of transitional, mesomerism-stabilized cations that react with excess Trp to yield 2-substituted Trp derivatives . The decomposition of Trp-metabolites could be induced by elevated or low pH-values that occur during the downstream processing of the Trp fermentation broth . IMT was detected in pharmaceutical-grade and feed-grade Trp in amounts of < 20-1,400 mg/kg . 1,1'-Ethylidenebis-(L-tryptophan) (EBT) is formed from acetaldehyde and Trp under acidic conditions and serves as a marker for EMS-suspicious Trp . 3-(Phenylamino)alanine (PAA) is the only not Trp derived case associated contaminant . Low amounts of PAA (20 mg/kg) could be detected in feed-grade Trp of one manufacturer . Non-EMS correlated 1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acids of Trp and formaldehyde, acetaldehyde and indole-3-acetaldehyde could be detected in the examined Trp raw materials (< 10-13,500 mg/kg) . In order to guarantee the safety of Trp containing drugs the amount of EBT (< 10 mg/kg Trp) and the sum of UV220 nm detectable contaminants (< 400 mg/kg Trp) are limited by the European authorities. J Ethnopharmacol, 2000 Apr, 70(1), 29 - 34 Nitric oxide-mediated endothelium-dependent relaxation of rat thoracic aorta induced by aqueous extract of red rice fermented with Monascus ruber; Rhyu MR et al.; Vasodilatory effects of aqueous extract of red rice fermented with Monascus ruber IFO32318 were examined on the isolated rat aorta . The water phase of fermented rice with Monascus (WP/FRM, 0.1-10 mg/ml) caused a transient relaxation of the endothelium-intact rat aorta precontracted with norepinephrine (NE, 300 nM) . The WP/FRM-induced relaxation was abolished by removal of endothelium or in the presence of N(G)-nitro-L-arginine (L-NNA, 10 microM), a nitric oxide (NO) synthase inhibitor . Neither atropine, a muscarinic receptor antagonist (10 microM), nor indomethacin, a cyclooxygenase inhibitor (10 microM), altered the WP/FRM-induced endothelium-dependent relaxation . gamma-Aminobutyric acid (GABA), one of the principle components of the extract, did not affect the muscle tension of the aorta with intact endothelium . In addition, WP/FRM increased the production of NO in primary cultured endothelial cells from human umbilical vein . The enhanced production of NO by WP/FRM was diminished by pretreatment with L-NNA (10 microM) . In conclusion, WP/FRM induces relaxation of rat aorta by releasing NO from endothelium . There seem to be some unknown factor(s) other than acetylcholine (Ach) and GABA, in the aqueous extract of red rice, which stimulate vascular endothelial cells to produce and/or release NO leading to endothelium-dependent relaxation by WP/FRM. Toxicol Lett, 2000 Mar 15, 112-113, 411 - 4 Effect of diet on colonic-programmed cell death: molecular mechanism of action; Chapkin RS et al.; Colon cancer evolves from a progressive inhibition of apoptosis and is influenced strongly by diet . Among dietary factors, butyrate (derived from fermentable fibers) may have utility as a chemopreventive agent because of its ability to promote apoptosis . Because CD95 (APO-1/Fas) transduces signals resulting in apoptosis, we tested the hypothesis that butyrate-dependent colonocyte apoptosis is mediated by this death receptor . Treatment of immortalized mouse colon cells with Fas agonistic antibody induced cell death, indicating that Fas in colonocytes is functional . Antagonism of Fas signaling using a soluble Fas:Fc chimera blocked butyrate induction of apoptosis . Therefore, Fas receptor dependent signal transduction is required for butyrate induction of apoptosis in colonic cells. Chin J Biotechnol, 1999, 15(2), 121 - 8 Studies on production of pullulan by the feed batch fermentation; Sun W et al.; The feed-batch fermentation of pullulan was investigated based on the conditions of batch fermentation . The optimal conditions of pullulan fermentation were determined by the investigation on the effect of feed mode, initial feed time, intermittent time, and composition of the feed solution on parameters such as productivity, conversion yield, biomass, dissolved oxygen (DO), pH of the fermentation broth, viscosity of the fermentation broth and molecular weight of the product . Above 70% of conversion yield and 100 kD of average molecular weight of the product was obtained at the optimal conditions. Chin J Biotechnol, 1999, 15(2), 105 - 11 Construction of a stable bioengineered strain of biotechmycin; Shang G et al.; A stable bioengineered strain of Biotechmycin (Streptomyces spiramyceticus WSJ-1) was constructed by integrating the 4"-isovaleryltransferase gene (ist) through homologous recombination into the chromosome of spiramycin-producing strain S.spiramyceticus F21 . In this construction, a Streptomyces/E.coli shuttle plasmid pKC1139 (AmR) was used as the vector and tsr gene was inserted as the marker for selection of homologous recombination . This constructed strain, S.spiramyceticus WSJ-1, was genetically very stable in production titer and in the production of biotechmycin as well as in carrying tsr selective marker when grown without pressure . The fermentation of S.spiramyceticus WSJ-1 was also improved compared with the original strain harboring unintegrated plasmid . Southern hybridization confirmed the integrated status of the ist gene in the host genome. Chin J Biotechnol, 1999, 15(2), 83 - 9 Purification and characterization of a novel fibrinolytic enzyme from Streptomyces spp; Wang J et al.; A novel protease with fibrinolytic activity, designated as SW-1, was isolated and purified from the fermentation broth of Streptomyces sp . strain Y405, a soil isolate . The purification procedure involved ammonium sulphate fractionation, decolorization on 290 resin, gel filtration on Sephadex G75, anion-exchange chromatography on DEAE Sephadex A25, and affinity chromatography on Lysine Sepharose 4B . About 4.2 mg purified enzyme was obtained from a liter of fermentation broth and the recovery yield was 12.0% . The purified enzyme showed the specific activity of 2952.3 urokinase units per milligram, which was increased by 230.6 fold over the fermentation broth . The purity determined by HPLC was 83.5% . SW-1 is a single chain polypeptide with a predicted molecular weight of 30 kDa in SDS-PAGE and an isoelectric point of 8.5 . The N-terminal sequence of SW-1 is R/N/F-P/D-G-M-T-M-T-A-I-A-N-Q-N-T-Q-I-N . There may be nonhomogeneity in the first and second amino acid residue of its N-terminal sequence . The analysis of amino acid composition showed that SW-1 consisted of 262 amino acids . The fibrinolytic activity of SW-1 was entirely inhibited by 10 mmol/L PMSF, 1 mmol/L EDTA and 1 mol/L lysine, respectively, suggesting that SW-1 is a serine protease and metalloprotease, and that the lysine binding site might play a role in the activity . The fibrinolytic activity of SW-1 is stable between 4-37 degrees C and pH 4.0-9.0, and the optimum pH is 8.0 . On plasminogen-free fibrin plates, SW-1 showed the same fibrinolytic activity as the mixture of SW-1 with plasminogen, indicating that SW-1 is a fibrinolytic enzyme which affects fibrin directly, but not a plasminogen activator which affects fibrin by activating plasminogen. Int J Food Sci Nutr, 1999 Jul, 50(4), 275 - 82 Evaluation of the nutritional quality and acceptability of fingermillet-based tempe as potential weaning foods in Tanzania; Mugula JK et al.; Six types of fingermillet (Eleusine coracana)-based tempe were developed by incorporating either commonbeans, groundnuts, cowpeas, mungbeans, chickpeas, sesame and/or their mixtures and fermented by Rhizopus oligosporus . The proximate and mineral composition was not changed significantly by fermentation . The protein content of tempe ranged from 13.3 to 15.7%; the total energy, 374.1 to 434.3 Kcal/100 g; the protein energy, 13.0 to 15.6%; the net dietary protein energy, 7.2 to 8.7%; and the chemical score, 61 to 71% . Tempe processing reduced the tannin and hydrogen cyanide content by 55.2 to 75.7, and 71.0 to 86.2%, respectively . It increased the content of reducing sugars, amino nitrogen, total acidity by 4 to 15, 3 to 6.9, and 4.3 to 12-fold, respectively . The in vitro protein digestibility was also improved . The developed tempe had protein quality and energy content recommended for weaning foods . The deep-fried tempe snacks were fairly organoleptically acceptable. Int J Food Sci Nutr, 1999 Jul, 50(4), 245 - 53 Intestinal degradation of dietary fibre in green beans--effects of microwave treatments; Svanberg M et al.; Intestinal degradation of dietary fibre in blanched and microwaved green beans was studied by using a rat experimental model . Content and composition of dietary fibre as well as molecular weight distribution of water-soluble polysaccharides (WSP) were analysed . There was a solubilization and a shift towards lower Mw of mainly uronic acid-containing polysaccharides with repeated microwave treatment in the raw material . Thus, the apparent Mw of water-soluble polysaccharides decreased from approximately 1,550,000 to approximately 300,000 . After the beans had been digested the Mw of the WSP was significantly reduced, to approximately 100,000, and the differences in Mw seen between various processed raw materials had been evened out . After fermentation the Mw of the WSP decreased further approximately 10 times . Fibre fermentability was high (approximately 90%) and similar for the various processed beans. Int J Food Sci Nutr, 1999 Sep, 50(5), 351 - 6 Calcium bioavailability of selected Egyptian foods with emphasis on the impact of fermentation and germination; Ghanem KZ et al.; The bioavailability of calcium (Ca) was assessed in 11 foodstuffs of plant or animal origin using rat feeding experiments and the criteria used for assessing the bioavailability were femur Ca and calcium efficiency . The bioavailability of Ca was found to be highest in fishes (Melouha) and cheeses (Mesh) fermented under local processing techniques . Germination of faba beans also enhanced the bioavailability of calcium to a mean value quite comparable to those of some dairy products, such as cottage cheese . The present study clearly demonstrates that the processes of fermentation and germination of selected foods are associated with an enhancement in the bioavailability of calcium . It is suggested that the breakdown of complex proteins under the fermentation or germination process is the underlying mechanism of action. Gut, 2000 Apr, 46(4), 507 - 14 Butyrate and trichostatin A effects on the proliferation/differentiation of human intestinal epithelial cells: induction of cyclin D3 and p21 expression; Siavoshian S et al.; BACKGROUND: Sodium butyrate, a product of colonic bacterial fermentation, is able to inhibit cell proliferation and to stimulate cell differentiation of colonic epithelial cell lines . It has been proposed that these cellular effects could be linked to its ability to cause hyperacetylation of histone through the inhibition of histone deacetylase . AIM: To analyse the molecular mechanisms of butyrate action on cell proliferation/differentiation and to compare them with those of trichostatin A, a well known inhibitor of histone deacetylase . METHODS: HT-29 cells were grown in the absence or presence of butyrate or trichostatin A . Cell proliferation and cell cycle distribution were studied after DNA staining by crystal violet and propidium iodide respectively . Cell cycle regulatory proteins were studied by western blot and reverse transcription-polymerase chain reaction . Cell differentiation was followed by measuring brush border enzyme activities . Histone acetylation was studied by acid/urea/Triton acrylamide gel electrophoresis . RESULTS: Butyrate blocked cells mainly in the G(1) phase of the cell cycle, whereas trichostatin A was inhibitory in both G(1) and G(2) phases . Butyrate inhibited the mRNA expression of cyclin D1 without affecting its protein expression and stimulated the protein expression of cyclin D3 without affecting its mRNA expression . Trichostatin A showed similar effects on cyclin D1 and D3 . Butyrate and trichostatin A stimulated p21 expression both at the mRNA and protein levels, whereas their effects on the expression of cyclin dependent kinases were slightly different . Moreover, butyrate strongly stimulated the activity of alkaline phosphatase and dipeptidyl peptidase IV, whereas trichostatin A had no effect . Finally, a six hour exposure to butyrate or trichostatin A induced histone H4 hyperacetylation . At 15 and 24 hours, histone H4 remained hyperacetylated in the presence of butyrate, whereas it returned to control levels in the presence of trichostatin A . CONCLUSIONS: The data may explain how butyrate acts on cell proliferation/differentiation, and they show that trichostatin A does not reproduce every effect of butyrate, mainly because of its shorter half life. J Food Prot, 2000 Mar, 63(3), 364 - 9 Inhibitory power of kefir: the role of organic acids; Garrote GL et al.; Milk and MRS broth fermented with kefir grains from different households were examined for inhibitory activity toward gram-negative and gram-positive strains . Fermented milk obtained with 10 g per 100 ml of inoculum (final pH 3.32 to 4.25) and MRS broth fermented with 1 and 10 g per 100 ml of inocula (final pH 4.18 to 5.25) had inhibitory power demonstrated by spot test and agar well diffusion assay . This inhibitory effect could be assigned to the undissociated form of lactic and acetic acid produced during the fermentation process . Kefir supernatants inhibited the growth of Escherichia coli 3 in nutrient broth at 37 degrees C for 24 h . However, supernatants of yogurt or milk artificially acidified with lactic and acetic acids allowed the growth of E . coli 3 in the same conditions . A bacteriostatic effect of milk fermented with kefir grains over E . coli 3 was also demonstrated. Plant Foods Hum Nutr, 1999, 54(3), 227 - 38 Proximate composition and selected functional properties of fermented and unfermented African oil bean (Pentaclethra macrophylla) seed flour; Akubor PI et al.; Flour samples were prepared from fermented and unfermented African oil bean (Pentaclethra macrophylla) seeds (AOBS) . The flour samples were evaluated for proximate composition and certain functional properties . The influence of defatting on these properties was also determined . Fermentation significantly increased (p < 0.05) the protein and decreased the crude fiber, ash, fat and carbohydrate contents of the AOBS flours . The nitrogen solubility of both fermented and unfermented flours was pH dependent with minimum and maximum solubility at pH 4.0 and pH 8.0, respectively, and with increased nitrogen solubility in the fermented sample . The fermented and unfermented flour samples had least gelation concentrations of 14 and 16% (w/v), respectively . The water absorption capacity and foam capacities of the fermented flour were 36 and 34%, respectively, over the unfermented seed flour . On the other hand, fermentation decreased the fat absorption capacity, emulsion activity and emulsion and foam stabilities . Fermentation decreased (p < 0.05) the bulk density of AOBS flour by 15% . Defatting improved all the functional properties evaluated except emulsion activity . These results indicate potential food uses of fermented and unfermented AOBS flour samples as protein supplements in diets and as functional ingredients in formulated foods. Plant Foods Hum Nutr, 1999, 54(3), 193 - 206 Production of African breadfruit (Treculia africana) and soybean (Glycine max) seed based food formulations, 1: Effects of germination and fermentation on nutritional and organoleptic quality; Ariahu CC et al.; Germination and fermentation were investigated as methods of improving the nutritional and organoleptic properties of soybean and African breadfruit seed based food formulations . Four products consisting of germinated-fermented soy-breadfruit seeds (GFSB), nongerminated-fermented soy-breadfrui