J Chromatogr A, 2001 Sep 21, 929(1-2), 23 - 9 Separation of endo-polygalacturonase using aqueous two-phase partitioning; Wu YT et al.; The partitioning of endo-polygalacturonase (endo-PG) in polyethylene glycol (PEG)-polyvinyl alcohol (PVA10000) and PEG-hydroxypropyl starch (Reppal PES100) aqueous two-phase systems was studied, and revealed the possibility of using aqueous two-phase extraction to purify and concentrate endo-PG from its clarified fermentation broth . For the PEG8000-PVA10000 system, endo-PG presented in the fermentation broth (at concentration that is more than 40% of total protein) mainly dominates in the top phase with a partitioning coefficient of 6, while total protein concentrates in the bottom phase . A separation scheme consisting of two consecutive aqueous two-phase extraction steps was proposed: a first extraction in polyethylene glycol (PEG8000)-polyvinyl alcohol system, followed by a second extraction in PEG8000-(NH4)2SO4 system . This allowed the separation of endo-PG from polymer and the recycling of PEG polymer, since endo-PG was very strongly partitioned into the bottom phase of the PEG8000-(NH4)2SO4 system . Laboratory-scale experiments were performed to test the efficiency of this scheme . It was found that enzyme recovery was up to 91% with a total purification factor of about 1.9 and a concentration factor of more than 5 . About 90% of the total PEG added into the systems can be recovered, and no reduction was obtained in the purification factor using recycled PEG. Reprod Nutr Dev, 2001 May-Jun, 41(3), 259 - 65 Utilisation of phytate phosphorus by rumen bacteria in a semi-continuous culture system (Rusitec) in lactating goats fed on different forage to concentrate ratios; Godoy S et al.; Experimental data on phytate phosphorus utilisation by ruminants are scarce . The aim of this study was to estimate the phytase activity of rumen micro-organisms when phytate phosphorus supply is high . A semi-continuous culture system fermentor (RUSITEC) was used . The inoculum was obtained from eight goats fed on either high or low forage level diets . Experimental buffers only differed by the nature of phosphorus monosodium phosphate vs . corn sodium phytate . The nylon bags containing 15 g DM of substrate were removed after a 48-hour incubation period . The system was maintained for 15 days: 5 days for adaptation, in order to obtain a steady state, and 10 days for sampling and recording . No significant differences were observed for DM digestibility, gas production, pH, N-NH3, and SCFA for the different treatments . Bacterial efficiency of phytate phosphorus utilisation was significantly higher (p < 0.001) with organic P, but remained lower than the data usually reported in the literature . These results may be explained by the relative saturation of bacterial phytase activity when the buffer contains a high level of phytate phosphorus. J Antibiot (Tokyo), 2001 Aug, 54(8), 635 - 41 Cladospolide D, a new 12-membered macrolide antibiotic produced by Cladosporium sp . FT-0012; Zhang H et al.; A new antibiotic termed cladospolide D was isolated along with the known cladospolides A and B from the fermentation broth of Cladosporium sp . FT-0012 by solvent extraction, ODS column chromatography and preparative HPLC . The structure of cladospolide D was deduced to be (E)-2-dodecen-5-hydroxy-11-olide-4-one . Cladospolide D showed antifungal activity against Pyricularia oryzae and Mucor racemosus. J Antibiot (Tokyo), 2001 Aug, 54(8), 628 - 34 TMC-205 a new transcriptional up-regulator of SV40 promoter produced by an unidentified fungus . Fermentation, isolation, physico-chemical properties, structural determination and biological activities; Sakurai M et al.; A new transcriptional up-regulator designated TMC-205 was discovered from the fermentation broth of an unidentified fungal strain TC 1630 by using an SV40 promoter-luciferase reporter assay . Based on spectroscopic analyses, its structure was determined to be (E)-6-(3-methyl-1,3-butadienyl)- H-indole-3-carboxylic acid . Expression of the luciferase activity was activated ca . 2-, 4-, and 6-fold by 1, 10, and 100 microM TMC-205, respectively . TMC-205 activated the transcriptional activity in a manner dependent on the presence of the enhancer element of SV40 in its promoter region. J Bacteriol, 2001 Nov, 183(21), 6466 - 77 Acetate and formate stress: opposite responses in the proteome of Escherichia coli; Kirkpatrick C et al.; Acetate and formate are major fermentation products of Escherichia coli . Below pH 7, the balance shifts to lactate; an oversupply of acetate or formate retards growth . E . coli W3110 was grown with aeration in potassium-modified Luria broth buffered at pH 6.7 in the presence or absence of added acetate or formate, and the protein profiles were compared by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Acetate increased the steady-state expression levels of 37 proteins, including periplasmic transporters for amino acids and peptides (ArtI, FliY, OppA, and ProX), metabolic enzymes (YfiD and GatY), the RpoS growth phase regulon, and the autoinducer synthesis protein LuxS . Acetate repressed 17 proteins, among them phosphotransferase (Pta) . An ackA-pta deletion, which nearly eliminates interconversion between acetate and acetyl-coenzyme A (acetyl-CoA), led to elevated basal levels of 16 of the acetate-inducible proteins, including the RpoS regulon . Consistent with RpoS activation, the ackA-pta strain also showed constitutive extreme-acid resistance . Formate, however, repressed 10 of the acetate-inducible proteins, including the RpoS regulon . Ten of the proteins with elevated basal levels in the ackA-pta strain were repressed by growth of the mutant with formate; thus, the formate response took precedence over the loss of the ackA-pta pathway . The similar effects of exogenous acetate and the ackA-pta deletion, and the opposite effect of formate, could have several causes; one possibility is that the excess buildup of acetyl-CoA upregulates stress proteins but excess formate depletes acetyl-CoA and downregulates these proteins. Br J Nutr, 2001 Oct, 86(4), 443 - 51 Ingestion of insoluble dietary fibre increased zinc and iron absorption and restored growth rate and zinc absorption suppressed by dietary phytate in rats; Hayashi K et al.; We examined the effects of ingestion of five types of insoluble fibre on growth and Zn absorption in rats fed a marginally Zn-deficient diet (6.75 mg (0.103 mmol) Zn/kg diet) with or without added sodium phytate (12.6 mmol/kg diet) . The types of insoluble fibre tested were corn husks, watermelon skin, yam-bean root (Pachyrhizus erosus) and pineapple core, and cellulose was used as a control (100 g/kg diet) . Body-weight gain in the cellulose groups was suppressed by 57 % by feeding phytate . Body-weight gain in phytate-fed rats was 80 % greater in the watermelon skin fibre and yam-bean root fibre group than that in the cellulose group . Zn absorption ratio in the cellulose groups was lowered by 46 and 70 % in the first (days 7-10) and second (days 16-19) measurement periods with feeding phytate . In the rats fed the phytate-containing diets, Zn absorption ratio in the watermelon skin, yam-bean root and pineapple core fibre groups was 140, 80 and 54 % higher respectively than that in the cellulose group, in the second period . Fe absorption was not suppressed by phytate, however, feeding of these three types of fibre promoted Fe absorption in rats fed phytate-free diets . The concentration of soluble Zn in the caecal contents in the watermelon skin fibre or yam-bean root fibre groups was identical to that in the control group in spite of a higher short-chain fatty acid concentration and lower pH in the caecum . These findings indicate that ingestion of these types of insoluble fibre recovered the growth and Zn absorption suppressed by feeding a high level of phytate, and factors other than caecal fermentation may also be involved in this effect of insoluble fibre. Mol Genet Genomics, 2001 Sep, 266(1), 48 - 55 Cytochrome c is not essential for viability of the fungus Aspergillus nidulans; Bradshaw RE et al.; The filamentous fungus Aspergillus nidulans is an obligate aerobe, which is capable of anaerobic survival, but not anaerobic growth . Since cytochrome c forms an essential part of the oxidative respiratory pathway it was expected that mutants lacking this component would be non-viable . Gene replacement of one homologue of the cycA (cytochrome c) gene was carried out in a diploid strain . Benomyl-induced haploidisation of this diploid yielded all cycA+ haploid colonies, initially suggesting that loss of cycA was indeed lethal . However, use of an alternative unbiased method to recover haploids yielded viable, but slow-growing, cycA- mutants . Replacement of the cycA locus in the cycA- mutants was verified by Southern blotting . Spectral analysis confirmed the absence of detectable levels of cytochrome c, and respiratory insensitivity to cyanide suggested the absence of cytochrome c-dependent respiration . Growth parameters were consistent with those expected of a CycA- mutant . Compared to the wild type, the mutants grew slowly on fermentable carbon sources, did not grow on non-fermentable carbon sources, and produced higher levels of ethanol . To our knowledge, this is the first report of a filamentous fungus that remains viable after complete elimination of a functional cytochrome c gene . We propose that the mutants are viable due to their ability to ferment and to use alternative respiratory pathways. Int J Food Microbiol, 2001 Sep 19, 69(1-2), 59 - 68 Taxonomical and technological characteristics of Saccharomyces spp . associated with blue veined cheese; Hansen TK et al.; In blue veined cheeses, the dominant yeast species in most cases is Debaryomyces hansenii . Saccharomyces spp . occurs less frequently, but they can be found in some blue veined cheeses . In the present study, the taxonomy of Saccharomyces spp . associated to blue veined cheeses was studied and comparisons made to type strains of Saccharomyces spp . and starter cultures of Saccharomyces spp . used in other food fermentations . Phenotypically, the cheese strains were referred to the Saccharomyces sensu stricto complex and were further identified as S . cerevisiae . Genotypically, the Saccharomyces spp . investigated were similar although chromosomal polymorphism were observed . Concerning the technological characteristics, they were similar in assimilation and fermentation of the residual sugars and organic acids naturally found in cheese . The investigated yeasts were also similar in their lipolytic activity being able to hydrolyse tributyrin and low chain (C:8), but not C:14 fatty acids . However, they differed in their tolerance to NaCl with the blue cheese strains showing a higher tolerance . The cheese strain S . cerevisiae FB 7 was the only yeast capable of degrading casein . It mainly degraded the alpha(s1)-casein and the beta(alpha2)-casein components . It was also the only isolate stimulating the development of Penicillium roqueforti in cheese agar imitating the conditions in blue veined cheese . The stimulation of P . roqueforti was most pronounced for the least proteolytic strain of P . roqueforti examined. Int J Food Microbiol, 2001 Sep 19, 69(1-2), 147 - 52 Occurrence and characterization of yeasts isolated from milks and dairy products of Apulia region; Corbo MR et al.; One hundred and five strains of yeasts isolated from milk of different animal origin and from typical Apulian cheeses were studied to identify and characterize yeast strains for further selection as starter cultures for cheese production . The most prevalent isolates belonged to the species Trichosporon cutaneum (15.24%), Candida catenulata (10.48%) and Yarrowia lipolytica (8.57%) . In order to evaluate the potential use as starter cultures, the occurrence of some selected properties, such as fermentation of glucose and lactose, assimilation of lactic acid, citric acid and lactose, growth at 4 degrees C and production of lipolytic and proteolytic enzymes in the strains belonging to the most frequent species was tested . C . catenulata and C . zeylanoides were positive for assimilation of lactic and citric acids and showed psychrotrophic aptitude . T . cutaneum was positive for all properties tested except for glucose and lactose fermentations . Y . lipolytica was endowed with remarkable lipolytic activity also at 4 degrees C and was positive for assimilation of lactic and citric acids, growth at 4 degrees C and proteolytic activity. Int J Food Microbiol, 2001 Sep 19, 69(1-2), 125 - 33 Some probiotic properties of yeast isolates from infant faeces and Feta cheese; Psomas E et al.; Yeast isolates from infant faeces and Feta cheese were characterized to species level by phenotypic criteria, Randomly Amplified Polymorphic DNA (RAPD)-PCR and mitochondrial DNA (mt-DNA) restriction analysis . Results suggested that there is a good agreement between phenotypic characterization of yeasts and RAPD-PCR at species level; in addition, RAPD-PCR as well as mt-DNA restriction analysis provided good discrimination at strain level . Some technological and probiotic properties of selected strains were also investigated . The test strains exhibited lipolytic and proteolytic activities . They also tolerated low pH and survived satisfactory in gastric juice in vitro as well as in the presence of bile . In general, the isolates from faeces were more resistant to low pH and bile than those from Feta cheese . Selected strains could be used as starter supplements for industrial fermentations. Biotechnol Prog, 2001 Sep-Oct, 17(5), 970 - 3 Changes in Saccharomyces cerevisiae development induced by magnetic fields; Motta MA et al.; Saccharomyces cerevisiae cultures exposed to 110 mT and 220 mT steady magnetic fields (SMF) were studied to observe eventual induced growth alterations and changes in metabolic activity . Cell mass (biomass) growth was evaluated by light spectrometry, and metabolic alterations were estimated on the basis of the CO2 pressure produced and by the culture media pH changes, measured at the beginning and the end of the observation . The yeast strain DAUFPE-1012, cultivated in a nonaerated liquid agar Sabouraud glucose medium, was exposed to SMF generated by NdFeBr magnets . Results showed alterations induced by 220 mT SMF as an increment in cell proliferation (1.84%) and an increased CO2 production (36.1%) as compared to control groups . Furthermore, the initial-to-final pH difference in 220 mT SMF exposed cultures was higher than the 110 mT SMF and the control values . The whole acidification and the rise in CO2 production observed after 220 mT SMF exposure did not correspond to the biomass growth values, as compared to the other cultures, and was apparently provoked by a enhancement in the cellular metabolic rate . This technique becomes very promising for future biotechnological applications in fermentative processes. Biotechnol Prog, 2001 Sep-Oct, 17(5), 887 - 92 Sugar recovery and fermentability of hemicellulose hydrolysates from steam-exploded softwoods containing bark; Boussaid A et al.; The hemicellulose sugar recovery and ethanol production obtained from SO2-catalyzed steam explosion of a mixed white fir (70%) and ponderosa pine (30%) feedstock containing bark (9% dry weight/dry weight) was assessed . More than 90% of the available hemicellulose sugars could be recovered in the hydrolysate obtained after steam explosion at 195 degrees C, 2.38 min, and 3.91% SO2, with 59% of the original hemicellulose sugars detected in a monomeric form . Despite this high sugar recovery, this hydrolysate showed low ethanol yield (64% of theoretical yield) when fermented with a spent sulfite liquor-adapted strain of Saccharomyces cerevisiae . In contrast, most hydrolysates prepared at higher steam explosion severity showed comparable or higher ethanol yields . Furthermore, the hydrolysates prepared from bark-free feedstock showed better fermentability (87% of theoretical yield) despite containing higher concentration of known inhibitors . The ethanol yield from the hydrolysate prepared from a bark-containing wood sample could be improved to 81% by an extra stage acid hydrolysis (121 degrees C for 1 h in 3% sulfuric acid) . This extra stage acid hydrolysis and steam explosion at higher severity conditions seem to improve the fermentability of the hydrolysates by transforming certain inhibitory compounds present in the hydrolysates prepared from the bark-containing feedstock and thus lowering their inhibitory effect on the yeast used for the ethanol fermentation. Biotechnol Prog, 2001 Sep-Oct, 17(5), 876 - 80 Production of a desulfurization biocatalyst by two-stage fermentation and its application for the treatment of model and diesel oils; Chang JH et al.; For the production of oil-desulfurizing biocatalyst, a two-stage fermentation strategy was adopted, in which the cell growth stage and desulfurization activity induction stage were separated . Sucrose was found to be the optimal carbon source for the growth of Gordonia nitida CYKS1 . Magnesium sulfate was selected to be the sulfur source in the cell growth stage . The optimal ranges of sucrose and magnesium sulfate were 10-50 and 1-2.5 g x L(-1), respectively . Such a broad optimal concentration of sucrose made the fed-batch culture easy, while the sucrose concentration was maintained between 10-20 g x L(-1) in the actual operation . As a result, 92.6 g x L(-1) of cell mass was acquired by 120 h of fed-batch culture . This cell mass was over three times higher than a previously reported result, though the strain used was different . The desulfurization activity of the harvested cells from the first stage culture was induced by batch cultivation with dibenzothiophene as the sole sulfur source . The optimal induction time was found to be about 4 h . The resting-cell biocatalyst made from the induced cells was applied for the deep desulfurization of a diesel oil . It was observed that the sulfur content of the diesel oil decreased from 250 mg-sulfur x L-oil(-1) to as low as 61 mg-sulfur x L-oil(-1) in 20 h . It implied that the biocatalyst developed in this study had a good potential to be applied to a deep desulfurization process to produce ultra-low-sulfur fuel oils. Environ Microbiol, 2001 Aug, 3(8), 502 - 11 Localization of processes involved in methanogenic degradation of rice straw in anoxic paddy soil; Glissmann K et al.; In anoxic paddy soil, rice straw is decomposed to CH(4) and CO(2) by a complex microbial community consisting of hydrolytic, fermenting, syntrophic and methanogenic microorganisms . Here, we investigated which of these microbial groups colonized the rice straw and which were localized in the soil . After incubation of rice straw in anoxic soil slurries for different periods, the straw pieces were removed from the soil, and both slurry and straw were studied separately . Although the potential activities of polysaccharolytic enzymes were higher in the soil slurry than in the straw incubations, the actual release of reducing sugars was higher in the straw incubations . The concentrations of fermentation products, mainly acetate and propionate, increased steadily in the straw incubations, whereas only a little CH(4) was formed . In the soil slurries, on the other hand, fermentation products were low, whereas CH(4) production was more pronounced . The production of CH(4) or of fermentation products in the separated straw and soil incubations accounted in sum for 54-82% of the CH(4) formed when straw was not removed from the soil . Syntrophic propionate degradation to acetate, CO(2) and H(2) was thermodynamically more favourable in the soil than in the straw fraction . These results show that hydrolysis and primary fermentation reactions were mainly localized on the straw pieces, whereas the syntrophic and methanogenic reactions were mainly localized in the soil . The percentage of bacterial relative to total microbial 16S rRNA content was higher on the straw than in the soil, whereas it was the opposite for the archaeal 16S rRNA content . It appears that rice straw is mainly colonized by hydrolytic and fermenting bacteria that release their fermentation products into the soil pore water where they are further degraded to CH(4) . Hence, complete methanogenic degradation of straw in rice soil seems to involve compartmentalization. Parasitology, 2001 Sep, 123(Pt 3), 315 - 24 The effect of dietary carbohydrates with different digestibility on the populations of Oesophagostomum dentatum in the intestinal tract of pigs; Petkevicius S et al.; An experiment was undertaken to study the effect of dietary carbohydrates with different digestibility on the populations of Oesophagostomum dentatum in the intestinal tract of pigs . Sixty-four worm-free pigs from a specific pathogen-free farm were randomly divided into 8 equal groups . The animals in 4 groups were assigned to a diet with partially undegradable carbohydrates (diet 1), while the pigs in the 4 remaining groups were given a diet with fermentable carbohydrates (diet 2) . Diet 1 was comprised of barley flour, oat husk meal, soybean meal, vitamins and minerals and diet 2 of barley flour, inulin and sugar beet fibre, soybean meal, vitamins and minerals . The pigs in 6 of the groups (n = 48) were inoculated with 6,000 infective larvae of O . dentatum . To determine O . dentatum populations at the early stage of infection, 16 pigs were slaughtered 3 weeks p.i., while the remaining 4 groups continued on the diets for a further 9 weeks after which they were slaughtered . In a diet cross-over experiment 6 weeks after inoculation, 8 pigs changed from diet 1 to diet 2 (diet 1 > diet 2), and 8 pigs from diet 2 to diet 1 (diet 2 > diet 1) . The results showed that partially undegradable carbohydrates provided favourable conditions not only for parasite establishment and sustainability, but also for already established O . dentatum infection while, in contrast, the diet composed of highly degradable carbohydrates decreased worm establishment, size and female fecundity . The implications for pastured pigs or pigs fed different complex carbohydrate diets is discussed. Microbiology, 2001 Oct, 147(Pt 10), 2749 - 56 Fermentable-sugar-level-dependent regulation of leukotoxin synthesis in a variably toxic strain of Actinobacillus actinomycetemcomitans; Inoue T et al.; Actinobacillus actinomycetemcomitans, a Gram-negative periodontopathic bacterium, produces a leukotoxin belonging to the RTX family . The production of leukotoxin varies greatly among different strains of this species and under different culture conditions . A toxin-production-variable strain, 301-b, stably produces significant amounts of leukotoxin in anaerobic fructose-limited chemostat cultures, but does not do so in the presence of excess fructose . This communication describes the cloning and sequencing of the leukotoxin promoter region from 301-b, showing that this strain has a promoter region similar to that from strain 652, a moderately toxic strain . Northern blot analysis using a leukotoxin gene probe demonstrated that change in toxin production in response to the level of external fructose was due to alteration in the transcriptional level of the leukotoxin gene . Pulsing of fructose into the fructose-limited chemostat culture remarkably reduced the intracellular cAMP level from 40 pmol (mg dry wt cells)(-1) to 3.1 pmol (mg dry wt cells)(-1), which was restored when the culture was returned to fructose-limited conditions . Further, it was found that addition of external cAMP to the culture with excess fructose resulted in an apparent recovery of leukotoxin production . Taken together, these findings indicate that a cAMP-dependent mechanism, possibly a catabolite-repression-like system, may be involved in the regulation of leukotoxin production in this bacterium. Carcinogenesis, 2001 Oct, 22(10), 1649 - 52 Wheat germ extract inhibits experimental colon carcinogenesis in F-344 rats; Zalatnai A et al.; It has been demonstrated for the first time that a wheat germ extract prevents colonic cancer in laboratory animals . Four-week-old inbred male F-344 rats were used in the study . Colon carcinogenesis has been induced by azoxymethane (AOM) . Ten rats served as untreated controls (group 1) . For the treatment of the animals in group 2, AOM was dissolved in physiologic saline and the animals were given three subcutaneous injections 1 week apart, 15 mg/kg body weight (b/w) each . In two additional groups Avemar (MSC), a fermented wheat germ extract standardized to 2,6-dimethoxy-p-benzoquinone was administered as a tentative chemo-preventive agent . MSC was dissolved in water and was given by gavage at a dose of 3 g/kg b/w once a day . In group 3, animals started to receive MSC 2 weeks prior to the first injection of AOM daily and continuously thereafter until they were killed 32 weeks later . In group 4 the basal diet and MSC were administered only . At the end of the experiment all the rats were killed by exsanguination, the abdominal large vessels were cut under a light ether anesthesia and a complete autopsy was performed . Percentage of animals developing colon tumors and number of tumors per animals: group 1 - 0 and 0; group 2- 83.0 and 2.3; group 3 - 44.8 (P < 0.001) and 1.3 (P < 0.004), group 4 - 0 and 0 . All the tumors were of neoplastic nature also histologically . The numbers of the aberrant crypt foci (ACF) per area (cm(2)) in group 2 were 4.85 while in group 3 the ACF numbers were 2.03 only (P < 0.0001). J Nutr Sci Vitaminol (Tokyo), 2001 Jun, 47(3), 253 - 7 Effect of dietary cabbage fermentation extract and young barley leaf powder on immune function of Sprague-Dawley rats; Miyazaki Y et al.; We investigated dietary effects of cabbage fermentation extract (CFE) and young barley leaf powder (YBLP) on rat immune functions . Male Sprague-Dawley rats of 4 wk age were fed for 3 wk diets containing these samples at 0.1 or 1% level . After the feeding period, serum IgG level was significantly higher in the rats fed 1% CFE . IgG productivity of spleen lymphocytes was enhanced dose-dependently in both groups of CFE and YBLP . Furthermore, IgG productivity of mesenteric lymph node (MLN) lymphocytes was approximately 2 times higher in the rats fed 1% CFE diet than in the control ones . IgA productivity of MLN lymphocytes tended to increase in both of CFE and YBLP groups . From these results, it was suggested that dietary CFE and YBLP reinforced Ig productivity in both systemic and intestinal immune systems . Moreover, CFE feeding tended to enhance the production of TNF-alpha by spleen lymphocytes . In spleen phospholipids, the level of arachidonic acid, a substrate for inflammatory lipid mediators, was not affected by CFE or YBLP feeding. J Dairy Sci, 2001 Sep, 84(9), 2066 - 72 Diurnal variation in pH reduces digestion and synthesis of microbial protein when pasture is fermented in continuous culture; de Veth MJ et al.; Many models of digestion assume steady-state conditions and do not account for diurnal variation in the rumen environment . This experiment examined the relationships between diurnal pH, pasture digestion, and microbial protein synthesis . Four dual-flow continuous culture fermenters were used to test the effect of increasing time at suboptimal pH on parameters of digestion . Fermentation of high quality pasture was controlled at pH 5.4 (suboptimal) for four intervals during each 24-h period (0, 4, 8, and 12 h) according to a 4 x 4 Latin square design . During the remainder of each day, pH was controlled at 6.3 (optimal) . Samples were collected during the last 3 d of each of the four 9-d experimental periods . A negative quadratic relationship was observed between time at suboptimal pH and apparent digestibility of organic matter and dry matter . The largest reduction in digestibility of organic matter, dry matter, and neutral detergent fiber was exhibited after 4 h at suboptimal pH . A negative linear relationship was found between time at suboptimal pH and microbial N flow, with the greatest decline in microbial N flow occurring at 12 h at suboptimal pH . These results suggest that the period of time that pH is below optimal may be more critical for digestion than the relationship between mean daily pH and optimal pH . Modeling non-steady-state ruminal conditions to account for diurnal variation in the ruminal environment may improve the prediction of digestion, especially fiber. J Chromatogr A, 2001 Aug 24, 927(1-2), 61 - 76 Affinity-reversed-phase liquid chromatography assay to quantitate recombinant antibodies and antibody fragments in fermentation broth; Battersby JE et al.; An automated dual-column liquid chromatography assay comprised of affinity and reversed-phase separations that quantifies the majority of antibody-related protein species found in crude cell extracts of recombinant origin is described . Although potentially applicable to any antibody preparation, we here use samples of anti-CD18 (Fab'2LZ) and a full-length antibody, anti-tissue factor (anti-TF), from various stages throughout a biopharmaceutical production process to describe the assay details . The targeted proteins were captured on an affinity column containing an anti-light-chain (kappa) Fab antibody (AME5) immobilized on controlled pore glass . The affinity column was placed in-line with a reversed-phase column and the captured components were transferred by elution with dilute acid and subsequently resolved by eluting the reversed-phase column with a shallow acetonitrile gradient . Characterization of the resolved components showed that most antibody fragment preparations contained a light-chain fragment, free light chain, light-chain dimer and multiple forms of Fab' . Analysis of full-length antibody preparations also resolved these fragments as well as a completely assembled form . Co-eluting with the full-length antibody were high-molecular-mass variants that were missing one or both light chains . Resolved components were quantified by comparison with peak areas of similarly treated standards . By comparing the two-dimensional polyacrylamide gel electrophoresis patterns of an Escherichia coli blank run, a production run and the material affinity captured (AME5) from a production run, it was determined that the AME5 antibody captured isoforms of light chain, light chain covalently attached to heavy chain, and truncated light chain isoforms . These forms comprise the bulk of the soluble product-related fragments found in E . coli cell extracts of recombinantly produced antibody fragments. J Ind Microbiol Biotechnol, 2001 Jun, 26(6), 363 - 8 An improved method for determination of ethyl carbamate in Korean traditional rice wine; Woo IS et al.; An improved extraction method for ethyl carbamate, a genotoxic and carcinogenic compound found in various fermented foods and beverages, was investigated for its determination in the two most typical Korean traditional rice wines, takju and yakju . When the rice wines were extracted twice with chloroform at 30 degrees C for 60 min, the recovery of ethyl carbamate was less than 16% . When they were saturated with NaCl before extraction, the recovery of ethyl carbamate increased to 24.4% in takju and 67.2% in yakju . Adjustment of pH to 9.0 after NaCl saturation in takju resulted in a dramatic increase of recovery to 81.2%, but not in yakju . When the contents of ethyl carbamate and its precursor, urea, in various Korean traditional rice wines were determined, there was no correlation between the two contents . This is due to the fact that storage time is more important than urea content in the formation of ethyl carbamate in rice wine . In addition, its storage at high temperature resulted in a dramatic increase in ethyl carbamate content according to the prolonged storage time, suggesting that storage time and temperature play a key role in the formation of ethyl carbamate in Korean traditional rice wine. Appl Environ Microbiol, 2001 Oct, 67(10), 4657 - 61 Cross-epithelial hydrogen transfer from the midgut compartment drives methanogenesis in the hindgut of cockroaches; Lemke T et al.; In the intestinal tracts of animals, methanogenesis from CO(2) and other C(1) compounds strictly depends on the supply of electron donors by fermenting bacteria, but sources and sinks of reducing equivalents may be spatially separated . Microsensor measurements in the intestinal tract of the omnivorous cockroach Blaberus sp . showed that molecular hydrogen strongly accumulated in the midgut (H(2) partial pressures of 3 to 26 kPa), whereas it was not detectable (<0.1 kPa) in the posterior hindgut . Moreover, living cockroaches emitted large quantities of CH(4) {105 +/- 49 nmol (g of cockroach)(-1) h(-1)} but only traces of H(2) . In vitro incubation of isolated gut compartments, however, revealed that the midguts produced considerable amounts of H(2), whereas hindguts emitted only CH(4) {106 +/- 58 and 71 +/- 50 nmol (g of cockroach)(-1) h(-1), respectively} . When ligated midgut and hindgut segments were incubated in the same vials, methane emission increased by 28% over that of isolated hindguts, whereas only traces of H(2) accumulated in the headspace . Radial hydrogen profiles obtained under air enriched with H(2) (20 kPa) identified the hindgut as an efficient sink for externally supplied H(2) . A cross-epithelial transfer of hydrogen from the midgut to the hindgut compartment was clearly evidenced by the steep H(2) concentration gradients which developed when ligated fragments of midgut and hindgut were placed on top of each other-a configuration that simulates the situation in vivo . These findings emphasize that it is essential to analyze the compartmentalization of the gut and the spatial organization of its microbiota in order to understand the functional interactions among different microbial populations during digestion. Br J Nutr, 2001 Sep, 86(3), 379 - 89 Content of short-chain fatty acids in the hindgut of rats fed processed bean (Phaseolus vulgaris) flours varying in distribution and content of indigestible carbohydrates; Henningsson AM et al.; Red kidney beans (Phaseolus vulgaris) processed to differ in distribution and content of indigestible carbohydrates were used to study hindgut fermentability and production of short-chain fatty acids (SCFA) . Bean flours with low or high content of resistant starch (RS), mainly raw and physically-inaccessible starch, were obtained by milling the beans before or after boiling . Flours containing retrograded starch and with a high or low content of oligosaccharides were prepared by autoclaving followed by freeze-drying with or without the boiling water . Six diets were prepared from these flours yielding a total concentration of indigestible carbohydrates of 90 or 120 g/kg (dry weight basis) . The total fermentability of the indigestible carbohydrates was high with all diets (80-87 %) . Raw and physically-inaccessible starch was more readily fermented than retrograded starch (97-99 % v . 86-95 %; ) . Non-starch glucans were fermented to a lesser extent than RS, but the fermentability was higher in the case of autoclaved (50-54 %) than boiled beans (37-41 %) . The distribution between acetic, propionic and butyric acid in the caecum was similar for all diets, with a comparatively high percentage of butyric acid (approximately 18) . However, with diets containing the high amounts of RS, the butyric acid concentration was significantly higher in the distal colon than in the proximal colon ( and for the high- and low-level diets respectively), whereas it remained constant, or decreased along the colon in the case of the other diets . Furthermore, the two diets richest in RS also promoted the highest percentages of butyric acid in the distal colon (24 and 17 v . 12 and 12-16 for the high- and low-level diets respectively). Br J Nutr, 2001 Sep, 86(3), 331 - 40 Comparison of non-tracer and tracer methods for determination of volatile fatty acid production rate in the rumen of sheep fed on two levels of intake; Martin C et al.; The aim of the present work was to estimate volatile fatty acid (VFA) production rate in the rumen of sheep fed two levels of intake using both a tracer (TM; by isotope dilution) and a non-tracer method (NTM; by supplementary infusion) in steady-state conditions . Six wethers received a diet containing 700 g lucerne hay and 300 g ground maize/kg in eight equal meals at 3 h intervals per d . The diet (9.8 MJ metabolizable energy (ME)/kg DM) was offered at 90 % ad libitum consumption (high intake, HI) or 45 % ad libitum consumption (low intake, LI) in a crossover design . Each sheep received five intrarumen VFA solutions infused continuously for 24 h at rates of 250 ml and 165 ml/h for the HI and LI respectively . The first infusion, considered as a control treatment (Con), consisted of a solution of {1-(13)C}propionate (7 mmol/d) . The four other solutions were isoenergetic (1.9 MJ ME/kg DM intake) mixtures of unlabelled propionate (C(3)) and butyrate (C(4)) at different levels: 0.90 mol C(4)/kg DM intake; 0.60 mol C(3)/kg DM intake; 0.30 mol C(3)/kg DM intake; 1.35 mol C(3)/kg DM intake . The VFA infusions did not affect rumen fermentation of the basal diet (pH, osmotic pressure, protozoa numbers), and comparable DM digestibility of the diet among the different treatments was observed . Both estimation methods demonstrated a similar increase (1.7-fold) in the rumen VFA production rate of sheep fed at intakes varying between 0.9 to 1.7 times maintenance . Irrespective of the intake level, the rumen production rate of individual VFA was on average 1.5-fold higher when estimated by the TM compared with the NTM . Rumen VFA production rates estimated by the NTM and TM represented 80 % and 120 % ME intake respectively . The difference between NTM and TM estimates seems likely to be caused mainly by overestimation of the VFA production rates by the TM. Protein Expr Purif, 2001 Oct, 23(1), 38 - 44 A large-scale purification of recombinant histone H1.5 from Escherichia coli; Pyo SH et al.; An Escherichia coli expression system has been constructed for production of biologically active recombinant histone H1.5 . A process of fermentation and purification method at a large scale has been developed . Recombinant histone H1.5 was released from the high density cultured cells by high-pressure homogenization . For an efficient removal of cell debris and partial purification of basic histone H1.5 in a single step, the whole cell lysates were directly loaded onto an expanded bed column packed with the strong cation exchanger (Streamline SP) . Complete removal of various impurities was achieved by a combination of hydroxyapatite chromatography and the following cation exchange chromatography with high grade strong cation exchanger (POROS 20 HS), and finally endotoxins were removed by ultrafiltration using a 100-kDa cut-off membrane, which gave the level of endotoxin below 0.5 EU/mg . The molecular mass of the recombinant histone H1.5 analyzed by MALDI-TOF-MS, and the N-terminal amino acid sequences were in good agreement with the authentic histone H1.5 . The whole process gave highly purified recombinant histone H1.5 at a high yield, compared to the conventional process . Anal Chem, 2001 Sep 1, 73(17), 4313 - 8 Optical biosensors . Monitoring studies of glycopeptide antibiotic fermentation using white light interference; Tunnemann R et al.; This paper describes the design, characterization, and use of an optical biosensor suited for the process control of biotechnological processes . The detector principle is based on reflectometric interference spectroscopy (RIfS) . RIfS enables a label-free, product-specific monitoring, with a future outline for on-line process control . The potential of the RIfS biosensor is exemplified by the qualitative and quantitative monitoring of the microbial production of vancomycin-type glycopeptide antibiotics. Arch Biochem Biophys, 2001 Oct 1, 394(1), 61 - 6 Urea increases tolerance of yeast inorganic pyrophosphatase activity to ethanol: the other side of urea interaction with proteins; Lopes DH et al.; Ethanol is the major product of yeast sugar fermentation and yet, at certain concentrations, it is very toxic to yeast cells . The major targets for ethanol's toxicity are the plasma membrane and the cytosolic enzymes: ethanol alters membrane organization and permeability and inactivates and unfolds globular cytosolic enzymes . The effects of ethanol on the plasma membrane are attenuated by the presence of trehalose, a disaccharide of glucose that is accumulated simultaneously with urea . The data presented in this paper show that trehalose is not effective at protecting yeast cytosolic inorganic pyrophosphatase against the inactivation of its catalytic activity promoted by alcohols . In contrast, 1 M trehalose increased the toxicity of alcohols against pyrophosphatase by at least 34% . On the other hand, 1.5 M urea attenuated the inactivation of pyrophosphatase promoted by alcohols by approximately 50% . Here we propose that, in the presence of alcohols, urea functions as a molecular filter, enriching the vicinity of the protein with water and excluding alcohol molecules . Conversely, trehalose tends to increase the interaction of alcohols with protein molecules, by withdrawing water, leading to a stronger inactivation promoted for a given concentration of alcohol in the bulk solution on pyrophosphatase activity . Int J Pharm, 2001 Oct 4, 227(1-2), 157 - 65 Colonic metabolism of ranitidine: implications for its delivery and absorption; Basit AW et al.; The aim of this study was to assess the in vitro stability of ranitidine to colonic bacteria by utilising a batch culture fermentation system to simulate the conditions of the colon . Three quantities of ranitidine, 100, 200 and 500 mg, in the form of the hydrochloride salt, were introduced into individual 100 ml fermenters consisting of buffer medium inoculated with freshly voided human faeces (10% w/v) . Control experiments were also run in parallel using equivalent drug quantities in buffer medium without the presence of faeces . Samples were removed at pre-determined time intervals over a 24 h period and were subsequently analysed by high-performance liquid chromatography (HPLC) for drug concentration . A selection of the samples removed from the fermenters was also analysed by conventional UV spectroscopy and mass spectrometry . Subsequent to an initial dissolution phase in the fermentation system, a marked decline in ranitidine concentration was noted over time, thereby suggesting degradation and metabolism of the drug by colonic bacteria . No such decline in concentration was noted in the control buffer systems . The rate and extent of metabolism was rapid and complete within 12 and 24 h for the 100 mg and 200 mg samples, respectively, although the largest sample size, 500 mg, was only partly metabolised over the course of the experiment . UV and mass spectrometry analysis indicated that metabolism occurred via cleavage of an N-oxide bond within the molecule with the resultant loss of an oxygen atom, although further metabolic reactions are possible . Such metabolism may in part be responsible for the poor bioavailability of ranitidine from the colon. Appl Biochem Biotechnol, 2001 Jun, 94(3), 213 - 23 Substrate selectivity of Gluconobacter oxydans for production of 2,5-diketo-D-gluconic acid and synthesis of 2-keto-L-gulonic acid in a multienzyme system; Ji A et al.; Substrate selectivity of Gluconobacter oxydans (ATCC 9937) for 2,5-diketo-D-gluconic acid (2,5-DKG) production was investigated with glucose, gluconic acid, and gluconolactone in different concentrations using a resting-cell system . The results show that gluconic acid was utilized favorably by G . oxydans as substrate to produce 2,5-DKG . The strain was coupled with glucose dehydrogenase (GDH) and 2,5-DKG reductase for synthesis of 2-keto-L-gulonic acid (2-KLG), a direct precursor of L-ascorbic acid, from glucose . NADP and NADPH were regenerated between GDH and 2,5-DKG reductase . The mole yield of 2-KLG of this multienzyme system was 16.8% . There are three advantages for using the resting cells of G . oxydans to connect GDH with 2,5-DKG reductase for production of 2-KLG: gluconate produced by GDH may immediately be transformed into 2,5-DKG so that a series of problems generally caused by the accumulation of gluconate would be avoided; 2,5-DKG is supplied directly and continuously for 2,5-DKG reductase, so it is unnecessary to take special measures to deal with this unstable substrate as it was in Sonoyama's tandem fermentation process; and NADP(H) was regenerated within the system without any other components or systems. Appl Biochem Biotechnol, 2001 Jun, 94(3), 197 - 211 Conversion of sodium lactate to lactic acid with water-splitting electrodialysis; Persson A et al.; The conversion of sodium lactate to lactic acid with water-splitting electrodialysis was investigated . One way of reducing the power consumption is to add a conductive layer to the acid compartment . Doing this reduced the power consumption by almost 50% in a two-compartment cell, whereas the electric current efficiency was not affected at all . Three different solutions were treated in the electrodialysis unit: a model solution with 70 g/L of sodium lactate and a fermentation broth that had been prefiltered two different ways . The fermentation broth was either filtered in an open ultrafiltration membrane (cut-off of 100,000 Dalton) in order to remove the microorganisms or first filtered in the open ultrafiltration membrane and then in an ultrafiltration membrane with a cut-off of 2000 Dalton to remove most of the proteins . The concentration of sodium lactate in the fermentation broth was 70 g/L, as well . Organic molecules present in the broth (peptides and similar organic material) fouled the membranes and, therefore, increased power consumption . Power consumption increased more when permeate from the more open ultrafiltration membrane was treated in the electrodialysis unit than when permeate from the membrane with the lower cut-off was treated, since there was a higher amount of foulants in the former permeate . However, the electrodialysis membranes could be cleaned efficiently with a 0.1 M sodium hydroxide solution. Bioresour Technol, 2001 Nov, 80(2), 149 - 51 Solid-state production of lignin peroxidase (LiP) and manganese peroxidase (MnP) by Phanerochaete chrysosporium using steam-exploded straw as substrate; Fujian X et al.; In the used media mainly consisting of steam-exploded wheat straw, the straw, which could replace expensive veratryl alcohol, might act not only as nutrient, but also as inducer of lignin enzymes . The activities of the enzymes lignin peroxidase (LiP) and manganese peroxidase (MnP) in solid-state fermentation (SSF) were far higher than in submerged fermentation (SmF) . Under optimal conditions of SSF, the maximum activities of the enzymes Lip and MnP were 2600 and 1375 U/L, respectively . Thus, this would pave the way for production and application of lignin enzymes on a large scale. J Biol Chem, 2001 Nov 23, 276(47), 43775 - 83 Epub 2001 Sep 18. NADP-glutamate dehydrogenase isoenzymes of Saccharomyces cerevisiae . Purification, kinetic properties, and physiological roles; DeLuna A et al.; In the yeast Saccharomyces cerevisiae, two NADP(+)-dependent glutamate dehydrogenases (NADP-GDHs) encoded by GDH1 and GDH3 catalyze the synthesis of glutamate from ammonium and alpha-ketoglutarate . The GDH2-encoded NAD(+)-dependent glutamate dehydrogenase degrades glutamate producing ammonium and alpha-ketoglutarate . Until very recently, it was considered that only one biosynthetic NADP-GDH was present in S . cerevisiae . This fact hindered understanding the physiological role of each isoenzyme and the mechanisms involved in alpha-ketoglutarate channeling for glutamate biosynthesis . In this study, we purified and characterized the GDH1- and GDH3-encoded NADP-GDHs; they showed different allosteric properties and rates of alpha-ketoglutarate utilization . Analysis of the relative levels of these proteins revealed that the expression of GDH1 and GDH3 is differentially regulated and depends on the nature of the carbon source . Moreover, the physiological study of mutants lacking or overexpressing GDH1 or GDH3 suggested that these genes play nonredundant physiological roles . Our results indicate that the coordinated regulation of GDH1-, GDH3-, and GDH2-encoded enzymes results in glutamate biosynthesis and balanced utilization of alpha-ketoglutarate under fermentative and respiratory conditions . The possible relevance of the duplicated NADP-GDH pathway in the adaptation to facultative metabolism is discussed. J Econ Entomol, 2001 Aug, 94(4), 892 - 7 Effect of a phloxine B-cucurbitacin bait on diabroticite beetles (Coleoptera: Chrysomelidae); Schroder RF et al.; Cucurbitacin E glycoside, extracted from a bitter mutant of Hawkesbury watermelon {Citrulls lanatus (Thunb.) Matsum . & Nakai (Syn . Citrullus vulgaris Schrad)} is the active ingredient of a feeding stimulant for the corn rootworm complex . It is the primary component of a water-soluble bait that can be combined with toxins for adult diabroticite control . Studies were conducted using phloxine B (D&C Red 28), a xanthene dye, as the toxin . This dye was efficacious against Diabrotica undecimpunctata howardi Barber, spotted cucumber beetle, and Acalymma vittatum (F.), striped cucumber beetle, in cucumber plots and could be recovered from cucumber leaves for 8 d after treatment . The average amount of dye recovered per dead spotted cucumber beetle at 8 d after treatment was 0.173 microg . Concentrated and sugar-free fermented forms of the watermelon extract were developed and compared with the fresh juice in field applications on cucumber plants . There was no significant difference in mortality of beetles from phloxine B-bait prepared with fresh, fermented, or concentrated extract, although in laboratory studies, fermented juice had higher feeding stimulant activity. Water Environ Res, 2001 May-Jun, 73(3), 276 - 85 Evaluation of the potential effects of equalization on the performance of biological phosphorus removal systems; Filipe CD et al.; Experimental data confirming that the phosphorus removal efficiency in biological excess phosphorus removal (BEPR) systems temporarily decreases when the amount of volatile fatty acids (VFAs) added in the anaerobic phase is suddenly increased are presented . This decrease in efficiency results from the fact that acetate uptake is a rapid process and that the phosphate concentration at the end of the anaerobic phase increases rapidly . Because of the nonlinear dependence of the phosphate uptake rate on the poly-beta-hydroxyalkanoate (PHA) content of phosphate-accumulating organisms (PAOs), the increase in PAO PHA content associated with VFA uptake is not able to cause a proportional increase in the rate of phosphate uptake . This causes a temporary imbalance between phosphate release and uptake, leading to lower phosphate removal efficiency . The VFA loading to full-scale BEPR systems is not constant throughout the day, and temporary imbalances such as the ones imposed in the batch tests can occur in full-scale systems . The effect of diurnal variations in loading was demonstrated through simulation of the behavior of an A/OTM system receiving a time-variable influent . Equalization is proposed as a method to diminish the potential for imbalances between phosphate release and uptake by avoiding sudden increases of VFA loading to the plant . Significant improvements in the effluent quality from the simulated system were achieved using equalization . The improvements were greater when the influent contained VFAs than when the VFAs were formed by fermentation in the anaerobic zone . The simulations suggested that it may be possible to decrease the amount of phosphorus discharged by a factor as high as 4 through use of concentration equalization . When both flow and concentration equalization were used, the total amount of phosphorus discharged was decreased by a factor of 8 . Equalization can be used, in concert with other strategies for preservation of the PHA content of PAOs under periods of low loadings, to minimize the magnitude of Monday phosphate peaks. Tree Physiol, 2001 Sep, 21(14), 1057 - 61 Changes in fine root production and longevity in relation to water and nutrient availability in a Norway spruce stand in northern Sweden; Majdi H; Effects of irrigation and liquid fertilization on fine root (< 1 mm) production and longevity, and fine root (< 0.5-2 mm) biomass were studied in a Norway spruce (Picea abies (L.) Karst.) stand in northern Sweden . Fine root length production and longevity were measured by the minirhizotron technique at 0-10 cm depth in the following treatments: irrigation (I), liquid fertilization (IL) and control (C) . Standing root biomass and root length density (RLD) were studied in the litter-fermented humus (LFH) layer and at depths of 0-10, 10-20 and 20-30 cm using soil cores in solid fertilized (F) and C plots . Minirhizotrons were installed in October 1994 and measurements recorded monthly from July to September 1995 and during the growing season in 1996 . Soil cores were sampled in 1996 . Fine root production increased significantly in IL plots compared with C plots, but the I treatment did not increase root production . Root mortality increased significantly in IL plots compared with C plots . Fine root longevity in IL plots was significantly lower compared with C and I plots . No significant difference was found between longevity of fine roots in I and C plots . Compared with C, F treatment increased fine root biomass in the LFH and mineral soil layers, and increased the amount of fine roots in mineral soil layers relative to the LFH layer . Furthermore, F increased RLD and the number of mycorrhizal root tips significantly. J Antibiot (Tokyo), 2001 Jul, 54(7), 554 - 61 Deacetylravidomycin M, a new inhibitor of IL-4 signal transduction, produced by Streptomyces sp . WK-6326 . I . Taxonomy, fermentation, isolation and biological activities; Arai M et al.; Streptomyces sp . WK-6326, a soil isolate, was found to produce an inhibitor of interleukin (IL)-4 signal transduction . Two structurally related compounds, a novel one designated deacetylravidomycin M and the known deacetylravidomycin, were isolated from the culture broth by solvent extraction, silica gel column chromatography and HPLC . Deacetylravidomycin M inhibited IL-4-induced CD23 expression in U937 cells without any cytotoxic effect, whereas deacetylravidomycin showed no inhibitory activity. J Antibiot (Tokyo), 2001 Jul, 54(7), 541 - 7 SNF4435C and D, novel immunosuppressants produced by a strain of Streptomyces spectabilis . I . Taxonomy, fermentation, isolation and biological activities; Kurosawa K et al.; SNF4435C and D, novel nitrophenyl pyrones, have been isolated from the culture broth of an actinomycete strain SNF4435 . The strain was identified as Streptomyces spectabilis from its morphological and cultural characteristics . SNF4435C and D showed a potent immunosuppressive activity in vitro and selectively suppressed B-cell proliferation induced by LPS versus T-cell proliferation induced by Con A. Planta, 2001 Aug, 213(4), 534 - 42 Metabolic changes associated with cluster root development in white lupin (Lupinus albus L.): relationship between organic acid excretion, sucrose metabolism and energy status; Massonneau A et al.; Under phosphorous deficiency, plants of white lupin (Lupinus albus L.) develop root clusters, which are also called proteoid roots due to their preferential presence in the Proteaceae . In their mature stage, these roots acidify the soil and excrete high amounts of carboxylates {up to 1.5 and 7 micromol (g FW)(-1) h(-1) of malate and citrate, respectively} enabling lupins to utilise sparingly available sources of phosphate . Using the amplified fragment length polymorphism (AFLP) technique, we identified genes predominantly expressed in juvenile and mature cluster roots . Transcripts for two enzymes involved in glycolysis, fructokinase and phosphoglucomutase, were identified in juvenile cluster roots and one, sucrose synthase, in mature cluster roots . In order to verify these observations we performed quantitative reverse transcription-polymerase chain reaction (RT-PCR) and could confirm the increased transcript level . Measurements of enzymatic activities showed that fructokinase and phosphoglucomutase activities increased in juvenile cluster roots, whereas sucrose synthase activity was maximal in mature cluster roots . These results indicate that formation of proteoid roots and citrate excretion increase sink strength locally . Production of citrate and inhibition of respiration are likely to result in an increased NADH/NAD+ ratio, which may be toxic for the plant . The fermentation pathway would allow oxidation of NADH by decarboxylation of pyruvate and subsequent reduction of the resulting acetaldehyde . Determination of alcohol dehydrogenase activity showed that this enzyme is strongly induced in mature proteoid roots . However, ethanol production was not increased, indicating that pyruvate is shunted to citrate synthesis and not to ethanol production. Lett Appl Microbiol, 2001 Sep, 33(3), 193 - 5 Mitomycin-supplemented washed blood agar for the isolation of Shiga toxin-producing Escherichia coli other than O157:H7; Sugiyama K et al.; AIMS: Isolation and recognition of the prominent Shiga toxin (Stx)-producing strains of Escherichia coli (STEC) serovar O157:H7 can be confirmed easily by their late fermentation of sorbitol and lack of beta-glucuronidase activity, but there has been no culture method of choice for detecting non-O157 STEC strains because of their biochemical diversity . Apart from Stx, many STEC strains produce enterohaemolysin (Ehly) regardless of their serovars . METHODS AND RESULTS: Although washed blood agar media, with or without the addition of antibiotics (vancomycin, cefixime, and cefsulodin) (WBA and WBVCCA), have been used to detect Ehly, a proportion of STEC strains consistently failed to produce haemolysin on these media . Washed blood agar medium was therefore studied further in order to increase the yield of strains producing Ehly . CONCLUSION: It was found that the addition of 0.5 microg ml(-1) of mitomycin C to the agar medium (WBMA) markedly increased the number of such strains . Thus, of 185 STEC strains comprising 95 O157 and 90 non-O157 STEC consisting of 34 serovars . Ninety-seven per cent of these strains produced haemolysis on WBMA, compared with only 76% and 83%, respectively, on WBA and WBVCCA . SIGNIFICANCE AND IMPACT OF THE STUDY: The appearance of the Ehly zone of haemolysis that was easily distinguishable from that of alpha-haemolysin was enhanced by the incorporation of mitimycin C into washed-blood medium. Bioresour Technol, 2001 Oct, 80(1), 63 - 71 Methane emission factors from cattle manure in Mexico; Gonzalez-Avalos E et al.; Factors responsible for methane emission from cattle manure representing diverse climates, systems and functions of cattle production are presented . These factors were obtained by means of an experimental methodology developed for this project . It was considered that the temperature, moisture, handling of manure and the animals' feed ration affect methane production . Drying conditions and fermentation of manure in cool, temperate and warm climates were simulated in the laboratory . Cattle manure was obtained from animals in intensive, semi-intensive and extensive production systems; for dairy, non-dairy and double purpose cattle production functions . Also handling of manure in dry lot, pasture and solid storage was considered . Results suggest that the dominant factor in methane emissions is the feed ration, followed by fermentation temperature and the excreta moisture content . The emission factors obtained in this work are at least a factor of five smaller than those proposed in the revised 1996 IPCC Guidelines for National Greenhouse Gas Inventories for countries like Mexico. Can J Diet Pract Res, 2000 Spring, 61(1), 24 - 26 Benefits of a Histamine-Reducing Diet for Some Patients with Chronic Urticaria and Angioedema; King W et al.; Urticaria and angioedema symptoms result primarily from the physiological actions of histamine . Some individuals with urticaria have a decreased ability to degrade dietary histamine before it enters the circulation . Foods high in histamine, such as fermented foods, may exacerbate urticaria and angioedema in these individuals . Certain food additives may increase endogenous release of histamine and urticaria and angioedema symptoms . The objective of this study was to evaluate the effect of a histamine-reducing diet on urticaria and angioedema symptoms, and on nutrient intake . Nineteen subjects with chronic urticaria or angioedema were randomized to a treatment group (n=9) or a control group (n=10) . The treatment group followed a histamine-reducing diet, and the control group eliminated artificial sweeteners from their diets . The subjects recorded antihistamine medication intake, number of wheals, the severity of pruritus and the severity of angioedema for two weeks before starting the diet and for six weeks during the dietary intervention . Subjects completed three-day food records every two weeks . There was a marginally significant decrease in the number of antihistamine tablets taken in the histamine-reducing diet group compared with the control group, and two of nine treatment subjects had dramatically improved symptoms . During the study there was no significant risk of nutritional deficiency for either group. FEMS Immunol Med Microbiol, 2001 Aug, 31(2), 163 - 7 Antitumor and antifungal activities in endophytic fungi isolated from pharmaceutical plants Taxus mairei, Cephalataxus fortunei and Torreya grandis; Huang Y et al.; The purpose of this work was to screen the endophytic fungi having antitumor or antifungal activity, which were isolated from the inner barks of three kinds of pharmaceutical plants, Taxus mairei, Cephalataxus fortunei and Torreya grandis, collected from Fujian province, China . Antitumor activity was studied by the MTT assay and antifungal activity was determined by observing fungal growth inhibition . 13.4% of endophytic fungi fermentation broths displayed cytotoxic activity on HL-60 cells at and below a dilution of 1:50, and 6.4% on KB cells . 52.3% of endophytic fungi fermentation broths displayed growth inhibition on at least one pathogenic fungi, such as Neurospora sp., Trichoderma sp . and Fusarium sp . Among all endophytic fungi isolated, the genus Paecilomyces sp . has the highest positive rate of antitumor and antifungal activity . These results indicate that endophytic fungi could be a promising source for antitumor and antifungal bioactive agents. Appl Microbiol Biotechnol, 2001 Aug, 56(3-4), 372 - 7 Production of the fungal biocontrol agent Ulocladium atrum by submerged fermentation: accumulation of endogenous reserves and shelf-life studies; Frey S et al.; A method was developed for the induction of submerged conidiation of Ulocladium atrum Preuss (isolate 385) for the first time, using an oatmeal extract broth . Two inoculum types were produced by this process: spores and mycelial fragments . Spore production was stimulated by reducing the broth water potential (psi) to -2.1 MPa and adding 20 mM calcium chloride . In contrast, mycelial fragments were dominant at -7.0 MPa psi . Maximum total inoculum (mycelial fragments and conidia) yields were approximately 2 x 10(7) ml(-1) after 9 days incubation at 25 degrees C at 100 rpm . Biomass from liquid cultures responded to water-stress by accumulating increased concentrations of endogenous sugar alcohols (polyols), particularly glycerol . Long-term shelf-life studies showed that submerged inoculum from cultures subjected to an intermediate water-stress (-2.1 MPa psi) and containing enhanced levels of glycerol (> 300 mg g(-1) freeze-dried material) retained viability significantly better (P < 0.05) than that from unstressed cultures, when assessed on agar with fully available water . This level of viability was comparable to that of aerial U . atrum spores from a 4-week solid-substrate fermentation on oat grains . However, in contrast to aerial spores, the ability of submerged biomass to germinate in drier conditions declined significantly after 6 months. Appl Microbiol Biotechnol, 2001 Aug, 56(3-4), 367 - 71 Production of the biocontrol agent Pantoea agglomerans strain CPA-2 using commercial products and by-products; Costa E et al.; The aim of this paper was to find the nitrogen and carbon sources that provide maximum biomass production of strain CPA-2 of the biocontrol agent Pantoea agglomerans and minimum cost of media, whilst maintaining biocontrol efficacy . To reduce the cost of media, commercial products and by-products were tested . P . agglomerans can be produced using a combination of nitrogen sources such as yeast extract (5 g l(-1)) and dry beer yeast (10 g l(-1)) with inexpensive carbohydrates such as sucrose (10 g l(-1)) and molasses (20 g l(-1)), respectively, maintaining the efficacy of the biocontrol agent against Penicillium digitatum and P . italicum on oranges . The results obtained in this study could be used to provide a reliable basis for a scale-up of this fermentation process to an industrial level. Appl Microbiol Biotechnol, 2001 Aug, 56(3-4), 289 - 95 Biotechnological production of itaconic acid; Willke T et al.; Itaconic acid (IA) is an unsaturated dicarbonic organic acid . It can easily be incorporated into polymers and may serve as a substitute for petrochemical-based acrylic or methacrylic acid . It is used at 1-5% as a co-monomer in resins and also in the manufacture of synthetic fibres, in coatings, adhesives, thickeners and binders . The favoured production process is fermentation of carbohydrates by fungi, with a current market volume of about 15,000 t/a . Due to the high price of about US$ 4/kg, the use of IA is restricted . At present, the production rates do not exceed 1 g l(-1) h(-1), accompanied by product concentrations of about 80 g l(-1) . New biotechnology approaches, such as immobilisation techniques, screening programmes and genetic engineering, could lead to higher productivity . Also, the use of alternative substrates may reduce costs and thus open the market for new and increased applications. J Ind Microbiol Biotechnol, 2001 Jan-Feb, 26(1-2), 15 - 21 Intracellular PHB conversion in a Type II methanotroph studied by 13C NMR; Vecherskaya M et al.; Poly-p-hydroxybutyrate (PHB) formation under aerobic conditions via incorporation of {13C-2}acetate as a cosubstrate and its intracellular degradation under anaerobic conditions in a Type II methanotroph was studied by 13C NMR . During PHB synthesis in the presence of labelled acetate, low levels of beta-hydroxybutyrate, butyrate, acetone, isopropanol, 2,3-butanediol and succinate were observed . Subsequent anaerobic PHB breakdown showed enhanced levels of these products at the expense of PHB . Fermentative metabolism occurring during anaerobic PHB degradation was confirmed in experiments with fully 13C-enriched cells, which were grown on 13C-labelled methane . beta-hydroxybutyrate, butyrate, acetate, acetone, isopropanol, 2,3-butanediol and succinate were detected as multiple 13C-labelled compounds in the culture medium . Our results suggest that intracellular PHB degradation can be used as a reserve energy source by methanotrophs under anoxic conditions. Adv Exp Med Biol, 2001, 488, 43 - 57 Electronic noses in food analysis; Haugen JE; Gas sensor array technology combined with multivariate data processing methods as artificial neural network has been demonstrated to have a promising potential for rapid non-destructive analysis of food quality . It may be applicable in quality control of raw material, food processing or products . This technique cannot completely replace reference methods like the use of sensory panels as the technique requires a frequent calibration against some valid reference method . As with all new techniques there remain some basic problems to be solved concerning sample handling and instrumental performance . The emerging research activity in the development of chemical sensors including hardware and software combined with applied research makes it realistic to expect applications with this technique implemented on-line in the food industry in near future . In particular, promising applications on meat seem to be within the field of spoilage, off-flavor, sensory analysis and fermentation processes. Adv Exp Med Biol, 2001, 488, 151 - 63 Solvent desorption dynamic headspace sampling of fermented dairy product volatiles; Rankin SA; A method was developed based on solvent desorption dynamic headspace analysis for the identification and relative quantification of volatiles significant to the study of fermented dairy product aroma . Descriptions of applications of this method are presented including the measurement of diacetyl and acetoin in fermented milk, the evaluation of volatile-hydrocolloid interactions in dairy-based matrices, and the identification of volatiles in cheeses for canonical discriminative analysis . Advantages of this method include rapid analysis, minimal equipment investment, and the ability to analyze samples with traditional GC split/splitless inlet systems . Limitations of this method are that the sample must be in the liquid state and the inherent analytical limitation to those compounds that do not coelute with the solvent or solvent impurity peaks. Water Sci Technol, 2001, 44(2-3), 77 - 83 Impact of solids management on nutrient ratios for in-line wastewater prefermenters; Rossle WH et al.; The full-scale evaluation of in-line prefermentation effects on wastewater nutrient ratios was performed under three different sludge operating conditions, with regards to sludge elutriation rates (0.7, 2.2 and 3.7 kg dry sludge/m3 raw sewage) and the corresponding sludge ages (17.3, 2.5 and 7.5 days) . At all three operating conditions the TKN/COD ratio increased (64, 46 and 20% respectively), with an average ratio of 0.057 mg N/mg COD in the raw sewage increasing to 0.082 mg N/mg COD in the settled sewage . These ratio increases can limit the use and performance of certain biological nutrient removal process configurations . This study has further highlighted the importance to counterbalance fermentation, thickening and solids removal requirements in a single tank in-line prefermenter . The minimum suspended solids removal (33%) was achieved at the lowest settled sludge solids content (4.3%), against a corresponding maximum volatile fatty acid generation rate (5.7 mg VFA/l/h) . The total solids and volatile fatty acid concentration profiles down the prefermenter tank depth were determined at a high sludge blanket level condition . It was demonstrated that the accumulated constituent mass inventory increase was about constant throughout the water and sludge layers respectively under such operational conditions. Water Sci Technol, 2001, 44(2-3), 343 - 50 Kaolinite sorption of Cd, Ni and Cu from landfill leachates: influence of leachate composition; Petrangeli Papini M et al.; Heavy metal speciation in landfill leachates plays a significant role in determining the mobility during the percolation through soils . The complexation characteristics of landfill leachate directly affects heavy metal solubility and the extent of the interaction with soils, lowering or raising the sorbed amount depending on the relative affinity of the complexed metal and uncomplexed form to soil adsorption sites . In this paper, the adsorption of Cd, Ni and Cu onto kaolinite from three leachates (collected from landfill at different fermentation stage) is studied, also in comparison with metal speciation by two different operative procedures . The heavy metals, at their natural concentration, were divided into operational classes according to an exchange-based procedure and by fractionation on the basis of molecular weight (exchange onto Chelex100 resin and ultrafiltration, respectively) . All the experiments were performed also on synthetic solutions designed according to leachate composition and theoretical speciation . The experimental results have shown leachate complexing capacity is strongly dependent on landfill age, and that broad parameters such as COD, DOC, pH, ionic strength and VFA concentration are not able to predict it . It is notheworthy that the strong complexing capacity of leachate can cause extraction of metals from the solid phase instead of adsorption from the liquid one. Can J Microbiol, 2001 Jul, 47(7), 685 - 9 Increased phospholipid transfer protein activity in Aspergillus oryzae grown on various industrial phospholipid sources; Asther M et al.; The effect of industrial carbon sources on phospholipid transfer protein production was investigated . Phospholipid fractions of different composition were prepared from various plant oils (i.e., soybean, rapeseed, and sunflower) according to the Lucas Meyer extraction and purification process . The effect of these fractions on phospholipid transfer protein activity of cell extracts from Aspergillus oryzae grown on medium containing these phospholipids as sole carbon source was studied . It was shown that phospholipid transfer activity was markedly increased by extracts containing a particular phospholipid composition . However, this stimulation depends mainly upon the phospholipid composition of the fraction used as fermentation substrate . Fractions enriched mainly in phosphatidylinositol (Epikuron 110), at the expense of phosphatidylcholine, were the most efficient sources for phospholipid transfer protein production by A . oryzae . Maximal phospholipid transfer activity, as well as biomass production, were increased 4.1- and 9.7-fold, respectively, when cultures were supplemented with Epikuron 110 prepared from sunflower lecithin, as compared to glucose-control cultures. Can J Microbiol, 2001 Jul, 47(7), 676 - 9 Debaryomyces hansenii growth in nonsterile seawater ClO2-peptone-containing medium; Ramirez-Orozco M et al.; We found that the marine yeast Debaryomyces hansenii strain C-11 (CIBNOR yeast collection, La Paz, Mexico) is highly tolerant to chlorine dioxide (ClO2), a powerful biocide agent . A direct application of this observation is the fermentation of the yeast in a nonsterile medium with an initial concentration of 0.3 mg/L of ClO2 . The disinfectant helps to avoid the growth of unwanted microorganisms while allowing the development of the yeast . Because the concentration of ClO2 decreases during the fermentation, we ascribe to D . hansenii cells a "biocontrol" action that contributes to the collection of a contaminant-free yeast cell biomass. Water Res, 2001 Oct, 35(14), 3441 - 7 Performance of UASB reactor treating leachate from acidogenic fermenter in the two-phase anaerobic digestion of food waste; Shin HS et al.; This study was conducted to investigate the performance of the upflow anaerobic sludge blanket (UASB) reactor treating leachate from acidogenic fermenter in the two-phase anaerobic digestion of food waste . The chemical oxygen demand (COD) removal efficiency was consistently over 96% up to the loading rates of 15.8 g COD/l d . The methane production rate increased to 5.51/l d . Of all the COD removed, 92% was converted to methane and the remaining presumably to biomass . At loading rates over 18.7 g COD/l d, the COD removal efficiency decreased due to sludge flotation and washout in the reactor, which resulted from short HRT of less than 10.6 h . The residual propionate concentration was the highest among the volatile fatty acids (VFA) in the effluent . The specific methanogenic activity (SMA) analysis showed that the VFA-degrading activity of granule was the highest for butyrate, and the lowest for propionate . Typical granules were found to be mainly composed of microcolonies of Methanosaeta . The size distribution of sludge particles indicated that partially granulated sludge could maintain the original structure of granular sludge and continue to gain size in the UASB reactor treating leachate from acidogenic fermenter. Semin Thromb Hemost, 2001 Aug, 27(4), 395 - 403 Recombinant von Willebrand factor: preclinical development; Plaimauer B et al.; Von Willebrand factor (vWF) is a multimeric glycoprotein (GP) that attracts platelets to the site of vascular injury, mediates platelet-platelet interaction, and stabilizes factor VIII (FVIII) in the circulation . Quantitative and qualitative defects of vWF result in von Willebrand disease (vWD), manifested by modest to severe bleeding episodes . Substitution therapy, with plasma-derived FVIII/vWF complex concentrates, is used for patients suffering the more severe forms of vWD . Efficacy of these preparations is often unsatisfactory because inadvertent proteolytic degradation during the manufacturing process causes them to lack the hemostatically most active high-molecular-weight multimers . In contrast, recombinant vWF (r-vWF), which is constitutively expressed at high yields in Chinese hamster ovary (CHO) cells and secreted into the conditioned medium under perfusion fermentation in "protein-free" medium, has high-molecular-weight multimers of extraordinary structural integrity . Functional analysis has shown that r-vWF promotes ristocetin cofactor-mediated platelet aggregation, collagen interaction and FVIII binding, and platelet-collagen adhesion under shear stress . Infusing vWF-deficient animals with r-vWF corrected vWF concentration and reduced blood loss, subsequently stabilizing endogenous FVIII associated with the reduction of bleeding time . Compared with plasma-derived vWF preparations, r-vWF was found to have a prolonged half-life, further enhancing the potential value of r-vWF as a therapeutic agent for treating patients suffering from vWD. Semin Thromb Hemost, 2001 Aug, 27(4), 385 - 94 Production processes of licensed recombinant factor VIII preparations; Boedeker BG; The state-of-the-art treatment for hemophilia A is replacement therapy with recombinant factor VIII (rFVIII) made possible by genetic engineering advances . Currently, there are four different products licensed and available for hemophilia A patients . All are produced by recombinant mammalian cells in large-scale fermenter cultures, purified to high purity, formulated in stable formulations and freeze dried . The first-generation products Recombinate and Kogenate (also sold as Helixate by Aventis) are characterized as full-length human factor VIII molecules and formulated using human serum albumin as a stabilizer . The second-generation product ReFacto contains an improved albumin-free sucrose formulation and incorporates advanced antiviral safety procedures in the manufacturing process . It is a truncated B region-deleted form of factor VIII (FVIII) that makes use of a nonhuman peptide linker 14 amino acids in length to connect the 80 and 90 kD subunits . The most recently licensed rFVIII product is the second-generation Kogenate product called KOGENATE Bayer/Kogenate FS, which combines the advantages of the human full-length FVIII molecule with an albumin-free, sucrose-based synthetic formulation as well as an improved viral safety profile . In this article, the manufacturing processes for each of the four different products are discussed in detail, focusing on expression systems and cell lines, culture medium, technical culture systems, purification process (including viral removal potential), and final formulation. Int J Food Microbiol, 2001 Aug 15, 68(1-2), 69 - 74 Surface mycoflora of a Spanish fermented meat sausage and toxigenicity of Penicillium isolates; Lopez-Diaz TM et al.; The surface mycoflora of "chorizo de Cantimpalos", a Spanish variety of fermented meat sausage characterised by a natural white covering, has been investigated . Among 54 mould strains isolated, 38 belonged to Penicillium subgenus Penicillium . The major species found (18 isolates) was identified as Penicillium commune, and the other dominant species (13 isolates) was identified as P . olsonii . None of the P . olsonii isolates produced cyclopiazonic acid, mycophenolic acid, roquefortine C . patulin or ochratoxin A, but all P . commune isolates produced cyclopiazonic acid . Toxicity to Artemia salina larvae was very high for all P . commune isolates investigated, while no isolates of P . olsonii studied were toxic to these crustaceans . The results may assist in selection of nontoxic strains, which could be used as surface starters in the manufacture of this type of sausage . The apparent inability to produce penicillin is a valuable characteristic to take into account in the selection process. Int J Food Microbiol, 2001 Aug 15, 68(1-2), 11 - 20 Comparison of culture, PCR and immunoassays for detecting Escherichia coli O157 following enrichment culture and immunomagnetic separation performed on naturally contaminated raw meat products; Chapman PA et al.; The aims of this study were (i) to evaluate the specificity and sensitivity of three previously described PCR assays for the detection of Escherichia coli O157 and, (ii) to compare PCR, culture, and two visual immunoassays (VIAs), BioSign and Path-Stik, for detecting E . coli O157 after enrichment culture and immunomagnetic separation (IMS) performed on various naturally contaminated raw beef, lamb and mixed meat products . Twelve sorbitol nonfermenting (SNF) verocytotoxin-producing (VT +) E . coli O157, 6 SNF VT - E . coli O157, 4 sorbitol fermenting (SF) VT + E . coli O157, 3 SF VT - E . coli O157, 23 E . coli belonging to 17 other serogroups and 12 organisms of other species were used to check the specificity of PCR reactions . Only one primer pair generated amplimers only with E . coli O157 and was used for all subsequent work . After enrichment culture and on inoculated minced beef samples, PCR was as sensitive as culture for detecting 9 of the 12 strains of E . coli O157, but up to 4 log10 more sensitive than culture for detecting three strains . Of the 120 samples of naturally contaminated meat products examined, 80 (67%) were positive by PCR, 70 (58%) were positive by BioSign, 69 (58%) were positive by culture and 67 (56%) were positive by Path-Stik . Although both VIAs lacked sensitivity when compared to PCR, both compared favourably with culture and both were extremely rapid and easy to perform, giving a result in less than 15 min . Eleven samples were positive by PCR and both VIAs, but negative by culture because culture plates were heavily overgrown with SF organisms, making detection of any E . coli O157 present impossible. J Bacteriol, 2001 Oct, 183(19), 5632 - 8 Rhamnose biosynthesis pathway supplies precursors for primary and secondary metabolism in Saccharopolyspora spinosa; Madduri K et al.; Rhamnose is an essential component of the insect control agent spinosad . However, the genes coding for the four enzymes involved in rhamnose biosynthesis in Saccharopolyspora spinosa are located in three different regions of the genome, all unlinked to the cluster of other genes that are required for spinosyn biosynthesis . Disruption of any of the rhamnose genes resulted in mutants with highly fragmented mycelia that could survive only in media supplemented with an osmotic stabilizer . It appears that this single set of genes provides rhamnose for cell wall synthesis as well as for secondary metabolite production . Duplicating the first two genes of the pathway caused a significant improvement in the yield of spinosyn fermentation products. Acta Astronaut, 1986 Mar, 13(3), 105 - 17 Gas transfer in microgravity fermentations; Thompson BG et al.; Fermentations performed under microgravity conditions may be used in future long duration space missions for recycling expendable life support materials . These fermentations will differ from similar fermentations performed at one gravity in the manner in which gas transfer in the fermentor is carried out. J Gravit Physiol, 1998 Jul, 5(1), P159 - 60 Effect of microgravitation on physiological-biochemical processes in orchids of different ecotypes; Cherevchenko TM et al.; NASA: A clinostat was used to study the metabolic processes of three types of orchids (Oncidium sphacelatum Lindl., Cattleya hybrida hort., and Zygopetalum mackaii Hook.) to determine genetic mechanisms of plant adaptation to microgravity . Metabolic aspects studied were the activity of oxidoreductive ferments in leaves (ascorbinoxidase, peroxidase, polyphenoloxidase, and cytochromoxidase) and the leaf content of chlorophylls and carotenoids . Life Support Biosph Sci, 1999, 6(1), 39 - 52 Food processing on a space station: feasibility and opportunities; Zasypkin DV et al.; An alternative strategy for processing plants into food on a space or other isolated station including an Advanced Life Support (ALS) system is proposed . Regular gravity (1 G) or hypogravity (< 1 G) has been considered . A key feature of this strategy is to include not only kitchen-scale preparation and processing but small-scale advanced food processing such as thermoplastic extrusion, homogenization, centrifugation, fermentation, etc . These processes are flexible and multifunctional and could significantly increase the variety, palatability, nutritional value, and shelf stability of foods, and the number of menu items based on ALS crops . The processes would minimize the time to process the food items and provide psychological support for the crew . The periodic processing of various crop harvests into shelf-stable foods for long-term storage can be performed . Unit operations as illustrated by various processing flow sheets on the manufacturing of individual products will be discussed in association with the equipment. Biotechnol Bioeng, 1985 Feb, 27(2), 151 - 5 Design of a system for the control of low dissolved oxygen concentrations: critical oxygen concentrations for Azotobacter vinelandii and Escherichia coli; Chen J et al.; The physiological activity of microorganisms in environments with low dissolved oxygen concentrations often differs from the metabolic activity of the same cells growing under fully aerobic or anaerobic conditions . This article describes a laboratory-scale system for the control of dissolved oxygen at low levels while maintaining other parameters, such as agitator speed, gas flowrate, position of sparger outlet, and temperature at fixed values . Thus, it is possible to attribute in dilute nonviscous fermentations all physiologic changes solely to changes in dissolved oxygen . Experiments were conducted with Azotobacter vinelandii and Escherichia coli . Critical oxygen concentrations for growth (that value of oxygen allowing growth at 97% of mu max) were measured as 0.35 +/- 0.03 mg/L for A . vinelandii and 0.12 +/- 0.03 mg/L for E . coli . These values are significantly different from the commonly quoted values for critical oxygen concentrations based on respiration rates . Because of the superior dissolved oxygen control system and an improved experimental protocol preventing CO2 limitation, we believe that the values reported in this work more closely represent reality. Econ Geol, 1985, 80, 270 - 82 Millimeter-scale variations of stable isotope abundances in carbonates from banded iron-formations in the Hamersley Group of Western Australia; Baur ME et al.; Several diamond drill cores from formations within the Hamersley Group of Western Australia have been studied for evidence of short-range variations in the isotopic compositions of the carbonates . For a set of 32 adjacent microbands analyzed in a specimen from the Marra Mamba Iron Formation, carbon isotope compositions of individual microbands ranged from -2.8 to -19.8 per mil compared to PDB and oxygen isotope compositions ranged from 10.2 to 20.8 per mil compared to SMOW . A pattern of alternating abundances was present, with the average isotopic contrasts between adjacent microbands being 3.0 per mil for carbon and 3.1 per mil for oxygen . Similar results were obtained for a suite of 34 microbands (in four groups) from the Bruno's Band unit of the Mount Sylvia Formation . Difficulties were experienced in preparing samples of single microbands from the Dales Gorge Member of the Brockman Iron Formation, but overall isotopic compositions were in good agreement with values reported by previous authors . Chemical analyses showed that isotopically light carbon and oxygen were correlated with increased concentrations of iron . The preservation of these millimeter-scale variations in isotopic abundances is interpreted as inconsistent with a metamorphic origin for the isotopically light carbon in the BIF carbonates . A biological origin is favored for the correlated variations in 13C and Fe, and it is suggested that the 13C-depleted carbonates may derive either from fermentative metabolism or from anaerobic respiration . A model is presented in which these processes occur near the sediment-water interface and are coupled with an initial oxidative precipitation of the iron. Adv Space Res, 1996, 18(1-2), 293 - 7 Anaerobic degradation of inedible crop residues produced in a Controlled Ecological Life Support System; Schwingel WR et al.; An anaerobic reactor seeded with organisms from an anaerobic lagoon was used to study the degradation of inedible crop residues from potato and wheat crops grown in a closed environment . Conversion of this biomass into other products was also evaluated . Degradation of wheat volatile solids was about 25% where that of potato was about 50% . The main product of the anaerobic fermentation of both crops was acetic acid with smaller quantities of propionate and butyrate produced . Nitrate, known to be high in concentration in inedible potato and wheat biomass grown hydroponically, was converted to ammonia in the anaerobic reactor . Both volatile fatty acid and ammonia production may have implications in a crop production system. Adv Space Res, 1996, 18(1-2), 251 - 65 Enzyme conversion of lignocellulosic plant materials for resource recovery in a Controlled Ecological Life Support System; Kohlmann KL et al.; A large amount of inedible plant material composed primarily of the carbohydrate materials cellulose, hemicellulose, and lignin is generated as a result of plant growth in a Controlled Ecological Life-Support System (CELSS) . Cellulose is a linear homopolymer of glucose, which when properly processed will yield glucose, a valuable sugar because it can be added directly to human diets . Hemicellulose is a heteropolymer of hexoses and pentoses that can be treated to give a sugar mixture that is potentially a valuable fermentable carbon source . Such fermentations yield desirable supplements to the edible products from hydroponically-grown plants such as rapeseed, soybean, cowpea, or rice . Lignin is a three-dimensionally branched aromatic polymer, composed of phenyl propane units, which is susceptible to bioconversion through the growth of the white rot fungus, Pluerotus ostreatus . Processing conditions, that include both a hot water pretreatment and fungal growth and that lead to the facile conversion of plant polysaccharides to glucose, are presented. Enzyme Microb Technol, 1993 Oct, 15, 899 - 900 Comments on "Bioprocessing in space" Volk T. An analysis developed by Westgate et al . for the digestible energy of edible and inedible biomass, including hydrolysis and fermentation, is reexamined with state-of-the-art values for the harvest index of hydroponic crops. Adv Space Res, 1989, 9(8), 179 - 84 Gas bubble coalescence in reduced gravity conditions; Thompson BG et al.; The effects of low gravity, as produced by a reduced gravity aircraft, the KC135, on the formation and coalescence of gas bubbles were examined over a range of gas-liquid ratios and with various medium constituents . These effects will influence design considerations of fermentors operating in reduced gravity conditions. Biotechnol Bioeng, 2001 Oct 20, 75(2), 212 - 8 Metabolic oscillations in an E . coli fermentation; Andersen DC et al.; Recombinant E . coli fermentations were observed to undergo regular, reproducible oscillations in oxygen uptake for several hours during a controlled fermentation process . Culture growth slowed during the period of oscillations, delaying induction of recombinant protein production . The oscillations were similar in 10-L and 1,000-L fermentors and also occurred with different feed control algorithms . Both observations support the hypothesis that the oscillations are metabolic in nature . Analysis of amino acid, ATP, and GTP pools suggests that the oscillations result from aberrant regulation of isoleucine biosynthesis leading to repeated starvation events in which protein synthesis and growth are impaired . Both a nutritional solution, isoleucine feeding, and a genetic solution, repair of an ilvG frameshift mutation in E . coli K-12 strains, were found to eliminate the oscillations, further supporting the proposed mechanism for the behavior . These results illustrate the interesting and complicated physiological behavior which can be displayed in metabolic networks and provide another example of surprising problems that can arise in growing recombinant organisms in fermentors . Biotechnol Bioeng, 2001 Oct 20, 75(2), 143 - 53 Economic comparison between conventional and disposables-based technology for the production of biopharmaceuticals; Novais JL et al.; Time to market, cost effectiveness, and flexibility are key issues in today's biopharmaceutical market . Bioprocessing plants based on fully disposable, presterilized, and prevalidated components appear as an attractive alternative to conventional stainless steel plants, potentially allowing for shorter implementation times, smaller initial investments, and increased flexibility . To evaluate the economic case of such an alternative it was necessary to develop an appropriate costing model which allows an economic comparison between conventional and disposables-based engineering to be made . The production of an antibody fragment from an E . coli fermentation was used to provide a case study for both routes . The conventional bioprocessing option was costed through available models, which were then modified to account for the intrinsic differences observed in a disposables-based option . The outcome of the analysis indicates that the capital investment required for a disposables-based option is substantially reduced at less than 60% of that for a conventional option . The disposables-based running costs were evaluated as being 70% higher than those of the conventional equivalent . Despite this higher value, the net present value (NPV) of the disposables-based plant is positive and within 25% of that for the conventional plant . Sensitivity analysis performed on key variables indicated the robustness of the economic analysis presented . In particular a 9-month reduction in time to market arising from the adoption of a disposables-based approach, results in a NPV which is identical to that of the conventional option . Finally, the effect of any possible loss in yield resulting from the use of disposables was also examined . This had only a limited impact on the NPV: for example, a 50% lower yield in the disposable chromatography step results in a 10% reduction of the disposable NPV . The results provide the necessary framework for the economic comparison of disposables and conventional bioprocessing technologies . Biotechnol Bioeng, 2001 Oct 5, 75(1), 63 - 73 The response of GS-NS0 myeloma cells to pH shifts and pH perturbations; Osman JJ et al.; The perceived sensitivity of animal cells to hydrodynamic shear has limited agitation and aeration at large-scale . This makes it difficult to ensure adequate mixing of the vessel contents and may lead to inhomogeneities in operational parameters such as temperature, dissolved oxygen concentration, and especially pH . The effect of pH shifts and pH perturbations on the cellular responses, in batch culture, of a GS-NS0 mouse myeloma cell line, expressing a recombinant antibody, was investigated . In addition, the effect of extreme pH on the structure of the purified antibody product was studied using isoelectric focusing . The fermentation pH value was shifted abruptly from pH 7.3 to pH values ranging from 6.5 to 9.0 . Culture pH was maintained at this new value for the remainder of the fermentation . All pH shifts of above 0.2 units caused a transient increase in apoptosis . However, cultures shifted to pH values between 7.0 and 8.0 continued to grow and the apoptotic fraction returned to initial levels . Cultures shifted to pH values above pH 8.0 and below pH 7.0 did not recover resulting in culture death . For example, a shift to pH 8.5 caused accumulation of cells in the G(2)/M phase of the cell cycle followed by apoptotic death . After the pH shift, maximum specific growth rate was observed over the range pH 7.3 to 7.5 and maximum viable cell number was seen at pH 7.3 . Maximum volumetric antibody production, resulting from increased culture longevity, was seen at pH 7.0 . It was also observed that glucose consumption increased with increasing pH . In a separate set of experiments cells were subjected to a single pH perturbation ranging in duration from 0 to 600 minutes . Exposure of cells to a pH value greater than 8.5 for more than 10 minutes caused a decrease in the proportion of viable cells and induced a lag in cell growth . At very low pH (6.5) similar effects were seen, but only for extended perturbations (600 min) . However, after recovery from the pH perturbation, growth, product secretion and metabolism all returned to original levels . Incubation of the antibody, at the range of pH values investigated, indicated no alterations in the structure of the antibody as determined by the isoelectric focusing pattern . Biotechnol Bioeng, 2001 Oct 5, 75(1), 39 - 45 Influence of temperature and pH on xylitol production from xylose by Debaryomyces hansenii; Converti A et al.; The production of xylitol from concentrated synthetic xylose solutions (S(o) = 130-135 g/L) by Debaryomyces hansenii was investigated at different pH and temperature values . At optimum starting pH (pH(o) = 5.5), T = 24 degrees C, and relatively low starting biomass levels (0.5-0.6 g(x)/L), 88% of xylose was utilized for xylitol production, the rest being preferentially fermented to ethanol (10%) . Under these conditions, nearly 70% of initial carbon was recovered as xylitol, corresponding to final xylitol concentration of 91.9 g(P)/L, product yield on substrate of 0.81 g(P)/g(S), and maximum volumetric and specific productivities of 1.86 g(P)/L x h and 1.43 g(P)/g(x) x h, respectively . At higher and lower pH(o) values, respiration also became important, consuming up to 32% of xylose, while negligible amounts were utilized for cell growth (0.8-1.8%) . The same approach extended to the effect of temperature on the metabolism of this yeast at pH(o) = 5.5 and higher biomass levels (1.4-3.0 g(x)/L) revealed that, at temperatures ranging from 32-37 degrees C, xylose was nearly completely consumed to produce xylitol, reaching a maximum volumetric productivity of 4.67 g(P)/L x h at 35 degrees C . Similarly, both respiration and ethanol fermentation became significant either at higher or at lower temperatures . Finally, to elucidate the kinetic mechanisms of both xylitol production and thermal inactivation of the system, the related thermodynamic parameters were estimated from the experimental data with the Arrhenius model: activation enthalpy and entropy were 57.7 kJ/mol and -0.152 kJ/mol x K for xylitol production and 187.3 kJ/mol and 0.054 kJ/mol x K for thermal inactivation, respectively . Biotechnol Bioeng, 2001 Oct 5, 75(1), 13 - 24 Intra-particle oxygen diffusion limitation in solid-state fermentation; Oostra J et al.; Oxygen limitation in solid-state fermentation (SSF) has been the topic of modeling studies, but thus far, there has been no experimental elucidation on oxygen-transfer limitation at the particle level . Therefore, intra-particle oxygen transfer was experimentally studied in cultures of Rhizopus oligosporus grown on the surface of solid, nutritionally defined, glucose and starch media . The fungal mat consisted of two layers--an upper layer with sparse aerial hyphae and gas-filled interstitial pores, and a dense bottom layer with liquid-filled pores . During the course of cultivation ethanol was detected in the medium indicating that oxygen was depleted in part of the fungal mat . Direct measurement of the oxygen concentrations in the fungal mat during cultivation, using oxygen microelectrodes, showed no oxygen depletion in the upper aerial layer, but revealed development of steep oxygen concentration gradients in the wet bottom layer . Initially, the fungal mat was fully oxygenated, but after 36.5 hours oxygen was undetectable at 100 microm below the gas-liquid interface . This was consistent with the calculated oxygen penetration depth using a reaction-diffusion model . Comparison of the overall oxygen consumption rate from the gas phase to the oxygen flux at the gas-liquid interface showed that oxygen consumption of the microorganisms occurred mainly in the wet part of the fungal mat . The contribution of the aerial hyphae to overall oxygen consumption was negligible . It can be concluded that optimal oxygen transfer in SSF depends on the available interfacial gas-liquid surface area and the thickness of the wet fungal layer . It is suggested that the moisture content of the matrix affects both parameters and, therefore, plays an important role in optimizing oxygen transfer in SSF cultures . Microbiology, 2001 Sep, 147(Pt 9), 2461 - 8 Energy metabolism of Actinobacillus actinomycetemcomitans during anaerobic and microaerobic growth in low- and high-potassium continuous culture; Ohta H et al.; Actinobacillus actinomycetemcomitans, a member of the gamma subclass of the Proteobacteria, has been implicated as the agent responsible for human periodontitis . In this study, A . actinomycetemcomitans 301-b was grown in fructose-limited chemostat cultures under anaerobic {redox potential (E(h))<-400 mV} and microaerobic (E(h)= -200 mV) conditions to characterize its energy metabolism . Effects of K(+) and Na(+) on growth and metabolism were also examined . In a control medium containing 5.2 mM K(+) and 24 mM Na(+), the molar growth yield on fructose (Y(fructose)) of microaerobic cultures was 1.3 times higher than the yield of anaerobic cultures at D < or =0.10 h(-1), but the difference in the Y(fructose) between microaerobic and anaerobic cultures decreased at D< or =0.10 h(-1) . When the ATP yield from fermentation was estimated from the amounts of fructose consumed and acetate formed, the value of the microaerobic culture (2.49 mol ATP produced per mol fructose consumed) was lower than the anaerobic value {3.13 mol ATP (mol fructose)(-1)} . Therefore, ATP production from fermentation could not account for the increase in the Y(fructose) at D > 0.10 h(-1) and thus additional ATP was expected to be generated via respiration . Assuming that the Y(ATP) (g cells formed per mol ATP synthesized) was similar between anaerobic and microaerobic cultures, the estimated ATP yield from respiration was between 1.2 and 2.0 mol ATP (mol fructose)(-1) below D=0.10 h(-1) and decreased to 0.3 mol ATP (mol fructose)(-1) when D was increased to 0.19 h(-1) . Such growth-rate-dependent decreases in the Y(fructose) and the estimated ATP production from respiration were also observed in a high-Na(+) (5.2 mM K(+) and 106 mM Na(+)) culture but not in a high-K(+) (81 mM K(+) and 24 mM Na(+)) culture . In the high-K(+) culture, the microaerobic Y(fructose) was 1.4-2.0 times higher than the anaerobic value and the respiration-derived ATP yield was estimated to be between 1.2 and 1.9 mol ATP (mol fructose)(-1) over a wide range of dilution rate . These results suggest that higher concentrations of extracellular K(+) are required for the respiration to occur in rapidly growing cells of A . actinomycetemcomitans. Microbiology, 2001 Sep, 147(Pt 9), 2437 - 46 Regulation of the ldhA gene, encoding the fermentative lactate dehydrogenase of Escherichia coli; Jiang GR et al.; The fermentative lactate dehydrogenase (LDH) of Escherichia coli is induced by low pH under anaerobic conditions . Both translational and transcriptional gene fusions to ldhA, which encodes the fermentative LDH, have now been made . Both types of ldhA-lacZ fusion were induced by low pH, but only in the absence of air . However, the translational fusions were consistently expressed at a five- to tenfold higher level than the transcriptional fusions, perhaps implying some post-transcriptional effect on ldhA expression . Introduction of arcB::Kan decreased expression of both translational and transcriptional ldhA-lacZ fusions by three- to fivefold . Disruption of mlc, which encodes a repressor of several genes of the phosphotransferase system, almost abolished expression of ldhA . Disruption of csrA caused a moderate drop in expression of both operon and protein ldhA fusions, whereas insertional inactivation of csrB or glgA had the opposite effect . These effects are probably indirect, resulting from alterations in sugar accumulation versus storage . Mutations in ptsG, cra, fnr, narL, rpoS, osmZ, appY, ack/pta, aceEF, pfl and ldhA had no effect on expression of the ldhA fusions . ldhA was not induced by the membrane-permeant weak acid benzoate, implying that it does not respond to the internal pH directly . Little pH induction was seen during growth on glycerol plus fumarate, suggesting that products of sugar fermentation are necessary for acid induction . Addition of succinate, acetate or lactate had no effect on ldhA expression . In contrast, pyruvate caused a two- to fourfold increase in expression of ldhA-lacZ . This accords with the idea that increased sugar metabolism indirectly induces ldhA. J Parasitol, 2001 Aug, 87(4), 714 - 7 Cultural and physiological observations on Trypanosoma rhodesiense and Trypanosoma gambiense . 1949; Tobie EJ et al.; 1 . A diphasic medium of simple preparation is described for the indefinite cultivation of T . rhodesiense and T . gambiense . 2 . The chief advantage of the medium is that it contains rabbit blood and thus obviates the necessity of using human blood . 3 . The flagellates develop only to the proventricular stage; hence the cultures are noninfective . 4 . The proventricular forms of both T . rhodesiense and T . gambiense consume sugar with the concomitant formation of acid . They are aerobic fermenters . 5 . Very little, if any, ammonia is produced by the living parasites. J Nutr, 2001 Sep, 131(9 Suppl), 2552S - 5S Industrial production of L-glutamine; Kusumoto I; The industrial production of L-glutamine (L-Gln) started with its fermentation in the late 1960s . Currently, it is manufactured for use as pharmaceuticals and health foods at the worldwide annual production of approximately 2000 metric tons . To manufacture high quality L-Gln at a low cost, it is of prime importance to obtain a strain of microorganism with good production efficiency and minimum by-products . Furthermore, to obtain the final crystalline powder product, the efficient removal of impurities contained in the fermentation broth becomes paramount . Therefore, the industrial process is designed to take into account characteristics of the fermentation broth as well as chemical, physical and biological properties of L-Gln . Points that should be considered in the process design and typical industrial production of L-Gln are described in this report. J Nutr, 2001 Sep, 131(9), 2248 - 51 Theaflavins in black tea and catechins in green tea are equally effective antioxidants; Leung LK et al.; Green tea catechins, including (-)-epicatechin (EC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin (EGC) and (-)-epigallocatechin gallate (EGCG), are oxidized and dimerized during the manufacture of black tea and oolong tea to form orange-red pigments, theaflavins (TF), a mixture of theaflavin (TF1), theaflavin-3-gallate (TF2A), theaflavin-3'-gallate (TF2B) and theaflavin-3,3'-digallate (TF3) . The present study was designed to compare the antioxidant activities of individual TF with that of each catechin using human LDL oxidation as a model . All catechins and TF tested inhibited Cu(+2)-mediated LDL oxidation . Analysis of the thiobarbituric acid-reactive substances (TBARS) and conjugated dienes produced during LDL oxidation revealed that the antioxidant activity was in the order: TF3 > ECG > EGCG > or = TF2B > or = TF2A > TF1 > or = EC > EGC . Four TF derivatives also demonstrated a dose-dependent antioxidant activity in Cu(+2)-mediated LDL oxidation at concentrations of 5-40 micromol/L . These results demonstrate that the TF present in black tea possess at least the same antioxidant potency as catechins present in green tea, and that the conversion of catechins to TF during fermentation in making black tea does not alter significantly their free radical-scavenging activity. Br J Cancer, 2001 Sep 1, 85(5), 661 - 7 Betel nut and tobacco chewing; potential risk factors of cancer of oesophagus in Assam, India; Phukan RK et al.; Cancer of the oesophagus is the most commonly diagnosed cancer in males in Assam, in north-eastern India, and ranks second for females . The chewing of betel nut, with or without tobacco and prepared in various ways, is a common practice in the region and a case-control study has been designed to study the pattern of risk associated with different ways of preparing and chewing the nuts . 358 newly diagnosed male patients and 144 female have been interviewed together with 2 control subjects for each case chosen at random from among the attendants who accompanied patients to hospital . There were significant trends in risk ratios associated with the frequency of chewing each day, with the duration of chewing in years and with the age at which the habit was started that were apparent for both males and females and which remained significant after allowance was made for other known risk factors, notably tobacco smoking and alcohol consumption . The adjusted ratios, in comparison with non-chewers, were 13.3 M and 5.7 F for chewing more than 20 times a day, 10.6 M and 7.2 F for persons who had chewed for more than 20 years and 10.3 M and 5.3 F for those who had started before the age of 20 . Among the different combinations of ingredients that were chewed the adjusted odds ratios were highest for those who had been using fermented betel nut with any form of tobacco (7.1 M and 3.6 F) . The risk associated with tobacco smoking and alcohol consumption, which are high in some parts of the world, were less in Assam than those associated with the chewing of betel nut . Org Lett, 2001 Sep 6, 3(18), 2811 - 4 Studies on the total synthesis of tallysomycin . Synthesis of the threonylbithiazole moiety containing a structurally unique glycosylcarbinolamide; Sznaidman ML et al.; {structure: see text} . Tallysomycins are glycopeptide antibiotics that were first isolated from fermentation broths of Streptoalloteichus hindustanus . They are structurally related to the bleomycins but contain an additional talose sugar attached via a unique glycosylcarbinolamide linkage . Herein we report the synthesis of a key tallysomycin intermediate that incorporates the glycosylcarbinolamide moiety unique to the tallysomycins. Int J Food Microbiol, 2001 Sep 1, 68(3), 173 - 86 Field survey and literature review on traditional fermented milk products of Ethiopia; Gonfa A et al.; The wide variety and the socio-economic and dietary importance of traditional fermented milk products of Ethiopia are discussed in this paper . Information on the microbiology of these products is sparse and has relevance to those organisms associated with spoilage and to those considered desirable for fermentation . There is a clear need to improve the production of African foods and beverages {Int . J . Food Microbiol . 18 (1993) 85} . The objective of this review was to document traditional technology used and information on the microbiology of the products, and to identify various constraints to the development and commercialisation of fermented milk products . Thereby the major problems and potential areas for improvement are pointed out . Ergo, the most important traditional product resembles yoghurt and, as the other traditional products, is prepared by "spontaneous" fermentation, commonly initiated by either "back slopping" or by repeated use of the same utensil . Other products include traditional fermented curd or ititu, trad