Fermentations

J Med Food, 2004 Fall, 7(3), 274 - 83
Pomegranate extracts potently suppress proliferation, xenograft growth, and invasion of human prostate cancer cells; Albrecht M et al.; We completed a multicenter study of the effects of pomegranate cold-pressed (Oil) or supercritical CO(2)-extracted (S) seed oil, fermented juice polyphenols (W), and pericarp polyphenols (P) on human prostate cancer cell xenograft growth in vivo, and/or proliferation, cell cycle distribution, apoptosis, gene expression, and invasion across Matrigel, in vitro . Oil, W, and P each acutely inhibited in vitro proliferation of LNCaP, PC-3, and DU 145 human cancer cell lines . The dose of P required to inhibit cell proliferation of the prostate cancer cell line LNCaP by 50% (ED(50)) was 70 microg/mL, whereas normal prostate epithelial cells (hPrEC) were significantly less affected (ED(50) = 250 g/mL) . These effects were mediated by changes in both cell cycle distribution and induction of apoptosis . For example, the androgen-independent cell line DU 145 showed a significant increase from 11% to 22% in G(2)/M cells (P <.05) by treatment with Oil (35 microg/mL) with a modest induction of apoptosis . In other cell lines/treatments, the apoptotic response predominated, for example, in PC-3 cells treated with P, at least partially through a caspase 3-mediated pathway . These cellular effects coincided with rapid changes in mRNA levels of gene targets . Thus, 4-hour treatment of DU 145 cells with Oil (35 microg/mL) resulted in significant 2.3 +/- 0.001-fold (mean +/- SEM) up-regulation of the cyclin-dependent kinase inhibitor p21((waf1/cip1)) (P <.01) and 0.6 +/- 0.14-fold down-regulation of c-myc (P <.05) . In parallel, all agents potently suppressed PC-3 invasion through Matrigel, and furthermore P and S demonstrated potent inhibition of PC-3 xenograft growth in athymic mice . Overall, this study demonstrates significant antitumor activity of pomegranate-derived materials against human prostate cancer.

J Biotechnol, 2004 Sep 30, 113(1-3), 211 - 30
Spectral analysis and fingerprinting for biomedia characterisation; Pons MN et al.; Classical culture media, as well as domestic and/or industrial wastewater treated by biological processes, have a complex composition . The on-line and/or in situ determination of some substances is possible, but expensive, as sample collection and pre-treatment are often necessary with strict rules of sterility . More global methods can be used to detect rapidly "accidents" such as the appearance of an undesirable by-product in a fermentation broth or of a toxic substance in wastewater . These methods combine a "hard" part, for sensing, and a "soft" part, for data treatment . Among potential "hard" candidates, spectroscopy can be the basis for non-invasive and non-destructive measuring systems . Some of them have been already tested in situ: ultra-violet-visible, infra-red (mid or near), fluorescence (mono-dimensional, two-dimensional or synchronous), dielectric, while others, more sophisticated, such as mass spectrometry, coupled or not to pyrolysis, nuclear magnetic resonance and Raman spectroscopy, have been proposed . All these methods provide spectra, i.e . large sets of data, from which meaningful information should be rapidly extracted, either for analysis or fingerprinting . The recourse to data-mining techniques (the "soft" part) such as principal components analysis, projection on latent structures or artificial neural networks, is a necessary step for that task . A review of techniques, mostly based on spectroscopy, with examples taken in the bioengineering field in general is proposed.

J Biotechnol, 2004 Sep 30, 113(1-3), 121 - 35
Stress in recombinant protein producing yeasts; Mattanovich D et al.; It is well established today that heterologous overexpression of proteins is connected with different stress reactions . The expression of a foreign protein at a high level may either directly limit other cellular processes by competing for their substrates, or indirectly interfere with metabolism, if their manufacture is blocked, thus inducing a stress reaction of the cell . Especially the unfolded protein response (UPR) in Saccharomyces cerevisiae (as well as some other yeasts) is well documented, and its role for the limitation of expression levels is discussed . One potential consequence of endoplasmatic reticulum folding limitations is the ER associated protein degradation (ERAD) involving retrotranslocation and decay in the cytosol . High cell density fermentation, the typical process design for recombinant yeasts, exerts growth conditions that deviate far from the natural environment of the cells . Thus, different environmental stresses may be exerted on the host . High osmolarity, low pH and low temperature are typical stress factors . Whereas the molecular pathways of stress responses are well characterized, there is a lack of knowledge concerning the impact of stress responses on industrial production processes . Accordingly, most metabolic engineering approaches conducted so far target at the improvement of protein folding and secretion, whereas only few examples of cell engineering against general stress sensitivity were published . Apart from discussing well-documented stress reactions of yeasts in the context of heterologous protein production, some more speculative topics like quorum sensing and apoptosis are addressed.

J Colloid Interface Sci, 2004 Nov 1, 279(1), 100 - 8
Crystallization of oxytetracycline from fermentation waste liquor: influence of biopolymer impurities; Li SZ et al.; Organic impurities in the fermentation broth of antibiotic production impose great difficulties in the crystallization and recovery of antibiotics from the concentrated waste liquor . In the present laboratory study, the inhibitory effect of biopolymers on antibiotic crystallization was investigated using oxytetracycline (OTC) as the model antibiotic . Organic impurities separated from actual OTC fermentation waste liquor by ultrafiltration were dosed into a pure OTC solution at various concentrations . The results demonstrated that small organic molecules with an apparent molecular weight (AMW) of below 10,000 Da did not affect OTC crystallization significantly . However, large biopolymers, especially polysaccharides, in the fermentation waste caused severe retardation of crystal growth and considerable deterioration in the purity of the OTC crystallized . Atomic force microscopy (AFM) revealed that OTC nuclei formed in the solution attached to the surfaces of large organic molecules, probably polysaccharides, instead of being surrounded by proteins as previously thought . It is proposed that the attachment of OTC nuclei to biopolymers would prevent OTC from rapid crystallization, resulting in a high OTC residue in the aqueous phase . In addition, the adsorption of OTC clusters onto biopolymers would destabilize the colloidal system of organic macromolecules and promote particle flocculation . OTC crystallization would therefore take place with the precipitation of abundant organic impurities . Hence, the removal of polysaccharides and other biopolymers by ultrafiltration can be an effective means of improving the recovery of OTC and similar antibiotics by crystallization from the fermentation waste.

Zhongguo Yi Xue Ke Xue Yuan Xue Bao, 2004 Aug, 26(4), 354 - 8
{Establishment of a drug screening model for identifying up-regulator of human high density lipoprotein receptor}; Liu XH et al.; OBJECTIVE: To establish a new drug screening model based on transcriptional regulation of human high density lipoprotein (HDL) receptor gene CD36 and LIMPII analogous-1 (CLA-1) for discovering up-regulator of this receptor . METHODS: The upstream regulatory sequence of CLA-1 was obtained by polymerase chain reaction . A recombinant reporter plasmid pGL3-CLAP was constructed by inserting the regulatory sequence upstream of luciferase gene of pGL3-Basic . Human hepatoma cell line BEL-7402 was transfected with pGL3-CLAP . Samples were detected by testing luciferase activity of transfected BEL-7402 cells in microtiter wells . RESULTS: The drug screening model was established and optimized . Significant difference was present between pGL3-CLAP and pGL3-Basic transfected BEL-7402 cells (P< 0.001), and coefficient of variation was less than 10% . After primary and secondary screening, 1 compounds and 3 fermentation extracts had up-regulating activities . CONCLUSION: This new drug screening model may be efficiently used to screen up-regulators of human HDL receptor expression, which might become lead compounds for new anti-atherosclerosis drugs.

Wien Klin Wochenschr, 2004 Jul 31, 116(14), 465 - 76
{Dietary fibre: more than a matter of dietetics . I . Compounds, properties, physiological effects}; Trepel F; Dietary fibre is a heterogeneous group of substances which have only one common characteristic: the non-digestability in the small bowel . With one exception all fibres are carbohydrates (poly- or disaccharides) . Some fibres are water-soluble, others are unsoluble . This property is associated with physiological effects . Soluble (viscous) fibres can bind water and thus form hydrocolloids or gels, unsoluble ones cannot . Dietary fibres play an essential role in the physiology of the gastrointestinal tract . They modify the absorption of nutrients (particularly carbohydrates and lipids) in the small bowel . They accelerate the gut transit time and determine stool composition and quantity . They are the main nutritional source for the colonic microflora . During the bacterial fermentation short-chain fatty acids are formed which are essential for nutrition and integrity of the colonocytes and for colonic function . Moreover gases, detoxicating enzymes, antioxidants and carcinogen-inactivating compounds arise . The most important fibres are cellulose, hemicellulose, pectin, guar, psyllium, beta-glucan, Klason lignin and digestion-resistant starch; they are present in varying amounts in plant foods and in fibre preparations . The usual daily intake of dietary fibre in Europe and the USA amounts to only 15-20 g, while health authorities and nutrition societies recommend a reference value of at least 30 g . Dietary fibres are applied as food-integrated, as supplement and as purified substances.

J Chromatogr A, 2004 Aug 6, 1045(1-2), 93 - 8
Integrated isolation of antibody fragments from microbial cell culture fluids using supermacroporous cryogels; Dainiak MB et al.; The present paper describes a chromatographic capture/purification step for the recovery of proteins directly from undiluted and unclarified cell culture broths using supermacroporous dimethylacrylamide (DMAA) cryogel . The interconnected character and the size (10-100 microm) of the pores of the adsorbent make it possible to process whole cell fermentation broths without blocking the column . Cu2+-iminodiacetic acid (IDA) DMAA cryogel has been used for the isolation and purification of excreted (His)6-tagged single chain (sc) Fv antibody fragments, (His)6-scFv, from E . coli cell culture . Bound protein was recovered with 0.2 M imidazole or with 20 mM EDTA and was practically cell-free . Chromatographic capture using Cu2+-IDA cryogel column was performed at flow rates of 300 and 600 cm/h, respectively and resulted in 84-96% recovery of (His)6-scFv fragments with a purification factor of 13-15 . The DMAA cryogel adsorbent is mechanically stable, can withstand harsh cleaning-in-place procedure and is relatively inexpensive . Chromatographic isolation of proteins using cryogels allows efficient removal of cells and can be operated at a flow rate as high as 600 cm/h . This novel technique has proven to be a scalable process, does not require special equipment and can be a good alternative to expanded bed adsorption and other integrated isolation techniques.

Bioprocess Biosyst Eng, 2004 Dec, 26(6), 353 - 9 Epub 2004 Sep 18.
Intelligent modelling of bioprocesses: a comparison of structured and unstructured approaches; Hodgson BJ et al.; This contribution moves in the direction of answering some general questions about the most effective and useful ways of modelling bioprocesses . We investigate the characteristics of models that are good at extrapolating . We trained three fully predictive models with different representational structures (differential equations, differential equations with inheritance of rates and a network of reactions) on Saccharopolyspora erythraea shake flask fermentation data using genetic programming . The models were then tested on unseen data outside the range of the training data and the resulting performances were compared . It was found that constrained models with mathematical forms analogous to internal mass balancing and stoichiometric relations were superior to flexible unconstrained models, even though no a priori knowledge of this fermentation was used.

J Dairy Sci, 2004 Oct, 87(10), 3473 - 9
Effects of substrate, passage rate, and pH in continuous culture on flows of conjugated linoleic acid and trans C18:1; Qiu X et al.; A dual-flow continuous culture system consisting of 4 fermenters was used in a 4 x4 Latin square design . The objective of the research was to evaluate the effects of solid dilution rate (SDR), pH, and concentration of linoleic acid (LA) in the feed mixture on the production of conjugated linoleic acid (CLA) and trans-C18:1 . The 4 treatments were 1) control = pH 6.5, 1% LA, 4%/h SDR; 2) high solid dilution rate (HSDR) = pH 6.5, 1% LA, 8%/h SDR; 3) high linoleic acid (HLA) = pH 6.5, 3% LA, 4%/h SDR; and 4) low pH (LPH) = pH 5.8, 1% LA, 4%/h SDR . Inoculum was collected 6 h after feeding from a cow fed 40% alfalfa hay and 60% grain . Liquid dilution rate was held at 0.12/h . All treatments except HLA contained 2% tallow . The LA was dissolved in buffer and continuously infused into the fermenters . The CLA flows were 16.5, 20.4, 23.2, and 25.2 mg/d for control, HSDR, HLA, and LPH, respectively . Compared with control, LPH increased flows of CLA, cis-C18:1, and C18:2, and decreased flow of C18:0 . The neutral detergent fiber (NDF) and acid detergent fiber (ADF) digestibilities were not affected by pH . The HSDR tended to increase CLA flow compared to control, possibly because a shorter solid retention time led to incomplete biohydrogenation (BH) . The NDF and ADF digestibilities and bacterial numbers were reduced by HSDR . With more LA available as a substrate for CLA, HLA resulted in a higher flow of CLA than control . The HLA resulted in the highest acid detergent fiber and fatty acid digestibilities, bacterial numbers, and BH . Increasing solids passage rate, reducing pH, and increasing dietary LA appears to increase in vitro CLA production.

Appl Microbiol Biotechnol, 2005 Feb, 66(5), 560 - 566 Epub 2004 Sep 16.
Effect of nitrogen limitation and surplus upon trehalose metabolism in wine yeast; Novo MT et al.; Trehalose metabolism in yeast has been related to stress and could be used as a stress indicator . Winemaking conditions are stressful for yeast and understanding trehalose metabolism under these conditions could be useful for controlling alcoholic fermentation . In this study, we analysed trehalose metabolism of a commercial wine yeast strain during alcoholic fermentation by varying the nitrogen levels from low (below adequate) to high (excess) . We determined trehalose, nitrogen, sugar consumption and expression of NTH1, NTH2 and TPS1 . Our results show that trehalose metabolism is slightly affected by nitrogen availability and that the main consumption of nitrogen occurs in the first 24 h . After this period, nitrogen is hardly taken up by the yeast cells . Although nitrogen and sugar are still available, no further growth is observed in high concentrations of nitrogen . Increased expression of genes involved in trehalose metabolism occurs mainly at the end of the growth period . This could be related to an adaptive mechanism for fine tuning of glycolysis during alcoholic tumultuous fermentation, as both anabolic and catabolic pathways are affected by such expression.

J Agric Food Chem, 2004 Sep 8, 52(18), 5612 - 20
Assessment of the production of antioxidants from winemaking waste solids; Cruz JM et al.; Winemaking waste solids (WS, resulting from red grapes after fermentation and distillation to recover spirits) were subjected to various processing schemes for isolating fractions with antioxidant activity . The liquors entrapped in WS as received were separated by pressing and freeze-dried to yield a fraction with antioxidant activity (measured as DPPH radical scavenging capacity) comparable to those of synthetic antioxidants . A second approach based on the direct processing of raw WS in sulfuric acid medium under fixed operational conditions and further extraction of hydrolysis liquors with ethyl acetate enabled the isolation of a fraction with higher antioxidant ability at an improved yield . The most favorable approach started with a washing stage leading to liquors (which were directly freeze-dried to yield 1.20 g of extract/100 g of oven-dry WS and presented an EC50 of 0.41 g of extract/L) and washed solids, which were dried and subjected to hydrolytic processing (i) with water as a reactive in an autocatalyzed reaction (autohydrolysis) or (ii) with sulfuric acid solutions to give an ethyl acetate-soluble fraction with improved antioxidant properties (EC50 in the range of 0.18-0.40 g/L) . Samples from washing liquors and processing of washed solids in aqueous medium were subjected to chromatographic fractionation and analysis to give isolates with remarkable antioxidant activity (with EC50 as low as 0.07 g/L) and to identify their major components.

J Agric Food Chem, 2004 Sep 8, 52(18), 5588 - 92
Rapid and sensitive automated method for glucose monitoring in wine processing; Serban S et al.; A rapid and sensitive automated method for glucose monitoring that might be employed during wine fermentation and processing was developed . A flow injection (FI) system coupled with an automated dilutor and the "redox-versatile" modified electrode were used to directly measure glucose in wine . To avoid interferences during wine analysis, different formulations of enzymatically modified carbon paste electrodes (CPE) were used and evaluated in oxidation and reduction mode . The best selectivity and sensitivity for glucose monitoring in real samples was obtained in cathodic mode at a fixed potential of 0 V versus Ag/AgCl using a CPE modified with glucose oxidase, horseradish peroxidase, and ferrocene as redox mediator . A total linear range of 0.02-50 g/L glucose was covered using this automated system and allowed the measurement of glucose in dry, medium, and sweet white or red wines without any sample pretreatment . The results showed a good correlation with the standard method, and the proposed method is very rapid, simple, and reliable and does not need skilled operators.

Scand J Gastroenterol, 2004 Jul, 39(7), 638 - 44
Effect of ethanol and some alcoholic beverages on gastric emptying in humans; Franke A et al.; BACKGROUND: There is a paucity of detailed and controlled studies on the action of ethanol and alcoholic beverages on gastric emptying in humans . This study was designed to compare the effect of beer, red wine, whisky and their comparable pure ethanol solutions on gastric emptying in a controlled and randomized investigation . METHODS: On separate days, 10 healthy, fasted subjects received the following solutions, in random order, through a gastric tube: 500 mL beer, red wine, comparable pure ethanol solutions (4% and 10% v/v), glucose (5.5% and 11.4% w/v) and water, 125 mL whisky and 40% (v/v) ethanol (both followed by 125 mL water) and 250 mL water . Gastric emptying of the test solutions was assessed using ultrasonography of the antrum . RESULTS: As measured by ultrasonography of the antrum, half emptying times of the ethanol solutions (4%, 10% and 40% v/v) were significantly (P < 0.05) longer (22.6 +/- 4.8, 22.7 +/- 4.3 and 27.8 +/- 3.3 min, respectively, n=10) than those of water (14.6 +/- 1.9 min (500 mL) and 13.2 +/- 1.7 min (250 mL), respectively) . The half emptying times of beer (39.3 +/- 4.3 min) and red wine (72.6 +/- 7.6 min) were significantly longer than those of the corresponding ethanol concentrations, whereas whisky was emptied at nearly the same rate (26.4 +/- 5.9 min) as 40% (v/v) ethanol . Emptying of glucose 5.5% and 11.4% (w/v) was significantly and dose dependently slower (29.7 +/- 4.5 and 64.8 +/- 8.9 min) than water . CONCLUSIONS: 1) Pure ethanol in concentrations of 4%, 10% and 40% (v/v) inhibits gastric emptying . 2) The inhibitory effect of beer and red wine, but not of whisky, is stronger than that of their comparable ethanol concentrations . 3) Caloric content and non-alcoholic ingredients in alcoholic beverages produced by fermentation (beer and wine), but not in those produced by distillation (whisky), are most likely responsible for this effect.

Public Health Nutr, 2004 Sep, 7(6), 765 - 72
Frequency of soy food consumption and serum isoflavone concentrations among Chinese women in Shanghai; Frankenfeld CL et al.; OBJECTIVE: The food-frequency questionnaire (FFQ) can be an efficient tool to evaluate dietary intake in large, population-based studies, especially for specific foods . The objective of this study was to validate the assessment of soy and isoflavone (daidzein and genistein) intakes, measured by an FFQ, by comparing intakes with serum isoflavone concentrations . DESIGN AND SETTING: Soy and isoflavone intakes and serum isoflavone concentrations were determined as part of a case-control study of dietary factors and risks of benign breast disease and breast cancer . The FFQ, administered during an in-person interview, included six soy-specific line items . Blood was drawn within one week of FFQ completion . SUBJECTS: In total, 1823 women living in Shanghai, People's Republic of China . RESULTS: In this population, soybean milk, fresh bean curd and other bean foods were eaten once per week, and fermented bean curd, fried bean curd puff and soybeans were eaten less than once per week . A significant linear trend (P<0.01) in serum isoflavone concentrations across increasing categories of soy and isoflavone intakes was observed, indicating that soy and isoflavone intakes, measured by the FFQ, well distinguished serum isoflavone concentrations . Linear trends were also observed in both case and control groups in stratified analyses, suggesting little differential bias by case-control status . CONCLUSIONS: The results suggest that the FFQ provides a useful marker of soy food consumption and isoflavone exposure in this population.

Environ Technol, 2004 Jun, 25(6), 647 - 55
Wet oxidation treatment of organic household waste enriched with wheat straw for simultaneous saccharification and fermentation into ethanol; Lissens G et al.; Organic municipal solid waste enriched with wheat straw was subjected to wet-oxidation as a pre-treatment for subsequent enzymatic conversion and fermentation into bio-ethanol . The effect of temperature (185-195 degrees C), oxygen pressure (3-12 bar) and sodium carbonate (0-2 g l(-1) ) addition on enzymatic cellulose and hemicellulose convertibility was studied at a constant wet oxidation retention time of 10 minutes . An enzyme convertibility assay at high enzyme loading (25 filter paper unit (FPU) g(-1) dry solids (DS) added) showed that up to 78% of the cellulose and up to 68% of the hemicellulose in the treated waste could be converted into respectively hexose and pentose sugars compared to 46% for cellulose and 36% for hemicellulose in the raw waste . For all wet oxidation conditions tested, total carbohydrate recoveries were high (> 89%) and 44-66% of the original lignin could be converted into non-toxic carboxylic acids mainly (2.2-4.5 % on DS basis) . Simultaneous saccharification and fermentation (SSF) of the treated waste at 10% DS by Saccharomyces cerevisae yielded average ethanol concentrations of 16.5 to 22 g 1(-1) for enzyme loadings of 5 and 25 FPU g(-1) DS, respectively . The cellulose to ethanol conversion efficiency during SSF was 50, 62, 65 and 70% for a total enzyme loading of 5, 10, 15 and 25 FPU g(-1) DS, respectively . Hence, this study shows that wet oxidation is a suitable pre-treatment for the conversion of organic waste carbohydrates into ethanol and that compatible conversion yields (60-65%) can be achieved at moderate enzyme loadings.

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi, 2004 Sep, 20(5), 575 - 7
{Purification of recombinant humanized anti-HBsAg Fab antibody by affinity chromatography}; Deng N et al.; AIM: To establish a steady purification method for producing recombinant humanized anti-HBsAg Fab from yeast fermentation supernatant . METHODS: Humanized anti-HBs scFv was used to immunize BALB/c mice to obtain anti-scFv monoclonal antibody (mAb) which was used to purify recombinant humanized anti-HBsAg Fab from yeast fermentation supernatant by affinity chromatography . RESULTS: The purity of purified recombinant anti-HBsAg Fab was above 95% and its recovery rate was about 75%-85% . CONCLUSION: An efficient affinity chromatography suitable for purification of recombinant humanized anti-HBsAg Fab in large-scale was established.

J Agric Food Chem, 2004 Sep 22, 52(19), 5950 - 6
Low molecular weight compounds responsible for savory taste of Indonesian soy sauce; Lioe HN et al.; Indonesian soy sauce is made using only soybeans as the nitrogenous source . Moromi obtained from fermentation of yellow soybeans using Aspergillus sojae as the starter was investigated . The fraction with molecular weights of less than 500 Da obtained by stepwise ultrafiltration was then fractionated by several chromatographic procedures, including gel filtration chromatography and RP-HPLC . Several chemical analyses, CE profiles, and taste profiles were performed to obtain the most intense umami fraction . The main components eliciting or enhancing the umami taste present in the fraction were purified and identified by protein sequencing, ESI-MS, and (1)H NMR at 400 MHz . Besides free l-glutamic acid and aspartic acid, free aromatic amino acids such as l-phenylalanine and l-tyrosine may also play an important role in impressing savory or umami taste of Indonesian soy sauce at their subthreshold concentrations and in the presence of salt and free acidic amino acids . This is reported as a new phenomenon of the so-called bitter amino acids.

Anal Chem, 2004 Sep 15, 76(18), 5498 - 502
Preparation, characterization, and application of an enzyme-immobilized magnetic microreactor for flow injection analysis; Nomura A et al.; Enzyme-immobilized magnetic microparticles (EMMP) have been prepared for use as a microreactor in flow injection analysis (FI) . The microparticles were directly injected into the FI system . Their retention occurred within the flow line by small permanent magnets located near the detector . The analytical utility of this concept was illustrated by the assay of glucose using glucose oxidase (GOx), immobilized microparticles, and amperometric detection of liberated hydrogen peroxide . The microparticles were derived from silica gel (nominal pore diameter, 15-80 nm) by impregnation with a citric acid/ethanol solution and a ferric nitrate/ethanol solution and then by calcination in a nitrogen atmosphere to produce ferrimagnetic fine particles of spinel-type iron oxide (gamma-Fe(2)O(3)) inside the pore . They were characterized by X-ray diffraction . The calibration curve of the glucose sample (2 microL injected) was linear between 2.5 x 10(-6) and 5 x 10(-4) mol/L (R = 0.9995), and the detection limit was 1.0 x 10(-6) mol/L or 0.36 ng of injected glucose (S/N = 3) . The repeatability for a 5 x 10(-4) mol/L glucose solution was RSD = 1.5% (n = 6) . Application to the assay of glucose in a fermentation broth is illustrated . The GOx MMP were stable and active for more than eight months when kept at 10 degrees C.

Transgenic Res, 2004 Jun, 13(3), 245 - 59
Recombinant protein expression plasmids optimized for industrial E . coli fermentation and plant systems produce biologically active human insulin-like growth factor-1 in transgenic rice and tobacco plants; Panahi M et al.; Human insulin-like growth factor-1 (hIGF-1) is a growth factor with clinical significance in medicine . The therapeutic potential of recombinant hIGF-1 (rthIGF-1) stems from the fact that hIGF-1 resembles insulin in many aspects of physiology . The expression of hIGF-1 in transgenic tobacco and rice plants using different expression cassettes is reported here . In the present study, two coding sequences were tested, one with the original human sequence, but partially optimized for expression in E . coli and the other with a plant-codon-optimized sequence that was expected to give a higher level of expression in plant systems . Three different hIGF-1 recombinant expression constructs were generated . All expression constructs utilized the maize ubiquitin 1 promoter with or without a signal sequence . Analyses conducted using a hIGF-1 specific ELISA kit showed all transgenic plants produced hIGF-1 and the accumulated hIGF-1 increased from the E . coli codon bias to higher levels when the hIGF-1 coding sequence was codon-optimized to match that of the maize zeamatin protein--the most transcribed gene in maize endosperm suspension cells . Further analyses that compared the functionality of the bacterial signal peptide Lam B in plants showed that this leader peptide led to lower expression levels when compared to transgenic plants that did not contain this sequence . This indicated that this expression construct was functional without removal of the bacterial signal sequence . The maize ubiquitin 1 promoter was found to be more active in rice plants than tobacco plants indicating that in this case, there was a class preference that was biased towards a monocot host . Biological analyses conducted using protein extracts from transgenic plants showed that the rthIGF-1 was effective in stimulating the in vitro growth and proliferation of human SH-SY5Y neuroblastoma cells . This indicated that the plant-produced rthIGF-1 was stable and biologically active . As some plants have been reported to express an endogenous insulin-like protein, we also looked for any effect of the human growth factor in transgenic plants, but no developmental or morphological differences with wild type tobacco or rice plants were detected . Since insulin and hIGF-1 share some overlapping roles, hIGF-1 may become a substitute therapeutic agent in subjects with certain defects in their insulin receptor signaling . Hence, if the full beneficial potential of rthIGF-1 is achieved, it is expected that in the future the demand will likely increase significantly.

Int J Food Microbiol, 2004 Oct 1, 96(1), 13 - 8
Determination of growth characteristics and lipolytic and proteolytic activities of Penicillium strains isolated from Argentinean salami; Ludemann V et al.; The growth of Penicillium spp . on the surface of meat-based dry fermented sausages provides them with a protective effect against some undesirable microorganisms . Penicillium also acts as an antioxidant, minimizes the risk of excessive drying, and it is responsible for flavor development due to the decomposition of proteins, free fatty acids and lactic acid . With the aim of developing starter cultures, important physiological properties such as growth and proteolytic and lipolytic activities were evaluated on 13 mold strains belonging to the genera Penicillium . These strains were isolated from Argentinean dry fermented meat sausages named "salami" . The selection was based on color, mycellium appearance and growth characteristics . The most important factors of the drying process of salami, such as temperature (14 and 25 degrees C), water activity (aw) (0.90, 0.95 and 1.00) and presence of 2.5% sodium chloride (NaCl), were analyzed . Although all strains analyzed were able to grow under the different conditions evaluated, they showed different growth velocity (K = mm/day) in response to temperature, aw and presence of NaCl in the media . All strains showed both proteolytic and lipolytic activities under the studied factors of the drying process . Nevertheless, differences in inter-species and even intra-species were found . The addition of NaCl gave a stimulant effect to the proteolytic activity at 25 degrees C, but the response at 14 degrees C was variable . The same variability was observed in the presence of salt, both at 25 and 14 degrees C, when the lipolytic activity was assayed . According to our results, detailed assays of the physiological capacities of indigenous strains proposed as starter cultures are required .

J Contam Hydrol, 2004 Oct, 74(1-4), 313 - 31
Simulated and experimental evaluation of factors affecting the rate and extent of reductive dehalogenation of chloroethenes with glucose; Lee IS et al.; Carbohydrates such as molasses are being added to aquifers to serve as electron donors for reductive dehalogenation of chloroethenes . Glucose, as a model carbohydrate, was studied to better understand the processes involved and to evaluate the effectiveness for dehalogenation of different approaches for carbohydrate addition . A simulation model was developed and calibrated with experimental data for the reductive dehalogenation of tetrachloroethene to ethene via cis-1,2-dichloroethene . The model included fermentors that convert the primary donor (glucose) into butyrate, acetate and hydrogen, methanogens, and two separate dehalogenator groups . The dehalogenation groups use the hydrogen intermediate as an electron donor and the different haloethenes as electron acceptors through competitive inhibition . Model simulations suggest first that the initial relative population size of dehalogenators and H(2)-utilizing methanogens greatly affects the degree of dehalogenation achieved . Second, the growth and decay of biomass from soluble carbohydrate plays a significant role in reductive dehalogenation . Finally, the carbohydrate delivery strategies used (periodic versus batch addition and the time interval between periodic addition) greatly affect the degree of dehalogenation that can be obtained with a given amount of added carbohydrate.

Curr Opin Biotechnol, 2004 Aug, 15(4), 349 - 55
Improved beta-lactam acylases and their use as industrial biocatalysts; Sio CF et al.; Whereas the beta-lactam acylases are traditionally used for the hydrolytic processing of penicillin G and cephalosporin C, new and mutated acylases can be used for the hydrolysis of alternative fermentation products as well as for the synthesis of semisynthetic beta-lactam antibiotics . Three-dimensional structural analyses and site-directed mutagenesis studies have increased the understanding of the catalytic mechanism of these enzymes . The yield of hydrolysis and synthesis has been greatly improved by process design, including immobilization of the enzyme and the use of alternative reaction media . Significant advances have also been made in the resolution of racemic mixtures by means of stereoselective acylation/hydrolysis using beta-lactam acylases.

J Appl Microbiol, 2004, 97(4), 792 - 801
New PCR-based methods for yeast identification; Hierro N et al.; AIMS: To characterize reference yeast strains and identify indigenous strains isolated from wine fermentations by PCR methods . METHODS AND RESULTS: We compared several PCR techniques for yeast identification . We used oligonucleotide primers that are complementary to (i) intron splice sites, (ii) REP and (iii) ERIC elements to produce PCR fingerprints that display specific patterns between the different yeast species . These three techniques were used to characterize 41 reference yeast strains belonging to 15 different species and to identify 40 indigenous strains isolated from grape must and wine fermentations . Species-specific banding patterns were obtained with the three PCR-techniques with different degrees of intraspecific differentiation depending on the method . By comparing the PCR fingerprints of unknown isolates with those produced by reference strains, we identified yeast strains isolated from an industrial wine fermentation . CONCLUSIONS: All three PCR techniques are rapid, reliable and simple methods of yeast identification . As far as we know, this is the first time that the primers designed for amplifying repetitive elements in bacteria have been successfully used in yeast . SIGNIFICANCE AND IMPACT OF THE STUDY: Industry needs rapid, reliable and simple methods of yeast identification . The proposed PCR techniques will allow to achieve this objective.

J Microbiol, 2004 Mar, 42(1), 51 - 5
Optimal fermentation conditions for enhanced glutathione production by Saccharomyces cerevisiae FF-8; Cha JY et al.; The influence of feedstock amino acids, salt, carbon and nitrogen sources on glutathione production by Saccharomyces cerevisiae FF-8 was investigated . Glucose, yeast extract, KH2PO4, and L-cysteine were found to be suitable feedstock . Highest glutathione production was obtained after cultivation with shaking for 72 h in a medium containing glucose 3.0% (w/v), yeast extract 3.0%, KH2PO4 0.06% and L-cysteine 0.06% . The glutathione concentration achieved using this medium increased 2.27-fold to 204 mg/l compared to YM basal medium.

BMC Genomics . 2004 Sep 08;5(1):63.
Integrated analysis of metabolic phenotypes in Saccharomyces cerevisiae; Duarte NC et al.; BACKGROUND: The yeast Saccharomyces cerevisiae is an important microorganism for both industrial processes and scientific research . Consequently, there have been extensive efforts to characterize its cellular processes . In order to fully understand the relationship between yeast's genome and its physiology, the stockpiles of diverse biological data sets that describe its cellular components and phenotypic behavior must be integrated at the genome-scale . Genome-scale metabolic networks have been reconstructed for several microorganisms, including S . cerevisiae, and the properties of these networks have been successfully analyzed using a variety of constraint-based methods . Phenotypic phase plane analysis is a constraint-based method which provides a global view of how optimal growth rates are affected by changes in two environmental variables such as a carbon and an oxygen uptake rate . Some applications of phenotypic phase plane analysis include the study of optimal growth rates and of network capacity and function . RESULTS: In this study, the Saccharomyces cerevisiae genome-scale metabolic network was used to formulate a phenotypic phase plane that displays the maximum allowable growth rate and distinct patterns of metabolic pathway utilization for all combinations of glucose and oxygen uptake rates . In silico predictions of growth rate and secretion rates and in vivo data for three separate growth conditions (aerobic glucose-limited, oxidative-fermentative, and microaerobic) were concordant . CONCLUSIONS: Taken together, this study examines the function and capacity of yeast's metabolic machinery and shows that the phenotypic phase plane can be used to accurately predict metabolic phenotypes and to interpret experimental data in the context of a genome-scale model.

Lett Appl Microbiol, 2004, 39(4), 369 - 75
Influence of cultivating conditions on the alpha-galactosidase biosynthesis from a novel strain of Penicillium sp . in solid-state fermentation; Wang CL et al.; AIMS: The work is intended to achieve optimum culture conditions of alpha-galactosidase production by a mutant strain Penicillium sp . in solid-state fermentation (SSF) . METHODS AND RESULTS: Certain fermentation parameters involving incubation temperature, moisture content, initial pH value, inoculum and load size of medium, and incubation time were investigated separately . The optimal temperature and moisture level for alpha-galactosidase biosynthesis was found to be 30 degrees C and 50%, respectively . The range of pH 5.5-6.5 was favourable . About 40-50 g of medium in 250-ml flask and inoculum over 1.0 x 10(6) spores were suitable for enzyme production . Seventy-five hours of incubation was enough for maximum alpha-galactosidase production . Substrate as wheat bran supplemented with soyabean meal and beet pulp markedly improved the enzyme yield in trays . CONCLUSIONS: Under optimum culture conditions, the alpha-galactosidase activity from Penicillium sp . MAFIC-6 indicated 185.2 U g(-1) in tray of SSF . SIGNIFICANT AND IMPACT OF THE STUDY: The process on alpha-galactosidase production in laboratory scale may have a potentiality of scaling-up.

J Dairy Res, 2004 Aug, 71(3), 279 - 87
Effects of feeding or abomasal infusion of canola oil in Holstein cows . 1 . Nutrient digestion and milk composition; Chelikani PK et al.; We determined the effects of feeding canola oil or infusing it into the abomasum on rumen fermentation, nutrient digestibility, duodenal flows of fatty acids, and milk composition in Holstein cows . Five ruminally and duodenally cannulated Holstein cows in late lactation were used in a 3 x 5 incomplete Latin square design . Treatments were 1) Control: basal diet (CON), 2) Control+supplementation of canola oil at 1 kg/d in the feed (FED), and 3) Control+abomasal infusion of canola oil at 1 kg/d (INF) . Compared with CON, feed intake, ruminal fermentation characteristics, ruminal and total tract digestibilities of nutrients were not significantly affected by FED treatment but duodenal flows and milk concentrations of fatty acids (FA) such as trans-11 18:1 and cis-9 trans-11 18:2 (conjugated linoleic acid, CLA) were increased . In contrast to the effects of FED, INF reduced feed intake, total VFA production, intestinal flows of nutrients, FA digestibility and yields of milk and milk fat . Both FED and INF significantly reduced the proportions of saturated and medium-chain FA, and increased cis 18:1 in milk . Concentrations of 18:2n-6 and 18:3n-3 in milk were increased nearly 2-fold with INF relative to CON . Dietary or postruminal supplementation of canola oil to late-lactation cows reduced saturated FA and increased unsaturated C18 in milk but nutrient digestion was adversely affected with abomasal infusion of canola oil.

Microbiol Mol Biol Rev, 2004 Sep, 68(3), 538 - 59, table of contents
Single-cell microbiology: tools, technologies, and applications; Brehm-Stecher BF et al.; The field of microbiology has traditionally been concerned with and focused on studies at the population level . Information on how cells respond to their environment, interact with each other, or undergo complex processes such as cellular differentiation or gene expression has been obtained mostly by inference from population-level data . Individual microorganisms, even those in supposedly "clonal" populations, may differ widely from each other in terms of their genetic composition, physiology, biochemistry, or behavior . This genetic and phenotypic heterogeneity has important practical consequences for a number of human interests, including antibiotic or biocide resistance, the productivity and stability of industrial fermentations, the efficacy of food preservatives, and the potential of pathogens to cause disease . New appreciation of the importance of cellular heterogeneity, coupled with recent advances in technology, has driven the development of new tools and techniques for the study of individual microbial cells . Because observations made at the single-cell level are not subject to the "averaging" effects characteristic of bulk-phase, population-level methods, they offer the unique capacity to observe discrete microbiological phenomena unavailable using traditional approaches . As a result, scientists have been able to characterize microorganisms, their activities, and their interactions at unprecedented levels of detail.

Biotechnol Bioeng, 2004 Sep 5, 87(5), 623 - 31
Arabidopsis glycosyltransferases as biocatalysts in fermentation for regioselective synthesis of diverse quercetin glucosides; Lim EK et al.; Regioselectivity of glycosyltransferases offers an important means to overcome the limitations of chemical synthesis of small molecule glycosides . In this study we explore a large multigene family of UDP-glucose:glycosyltransferases of Arabidopsis for their potential as novel biocatalysts for in vitro synthesis and whole-cell catalysis . We used quercetin as a substrate for this study because the flavonol and its glycosides have important medicinal properties and the metabolite provides a complex structure for regioselective glucosylation . We analyzed the activity of 91 recombinant enzymes for in vitro activity toward quercetin and discovered 29 that are capable of glucosylating the substrate . We demonstrate the first enzymic synthesis of a range of glucosides in vitro, including the 3-O-, 7-O-, 3'-O-, and 4'-O-monoglucosides, 3,7-di-O-glucoside, and 7,3'-di-O-glucoside . We also show that the regioselectivity of glucosylation can be maintained when the enzymes are used as whole-cell biocatalysts in Escherichia coli.

Biotechnol Bioeng, 2004 Sep 5, 87(5), 593 - 601
Real-time update of calibration model for better monitoring of batch processes using spectroscopy; Kornmann H et al.; In order to reduce the large calibration matrix usually required for calibrating multiwavelength optical sensors, a simple algorithm based on the addition in process of new standards is proposed . A small calibration model, based on 14 standards, is periodically updated by spectra collected on-line during fermentation operation . Concentrations related to these spectra are reconciled into best-estimated values, by considering carbon and oxygen balances . Using this method, fructose, acetate, and gluconacetan were monitored during batch fermentations of Gluconacetobacter xylinus 12281 using mid-infrared spectroscopy . It is shown that this algorithm compensates for noncalibrated events such as production or consumption of by-products . The standard error of prediction (SEP) values were 0.99, 0.10, and 0.90 g/L for fructose, acetate, and gluconacetan, respectively . By contrast, without an updating of the calibration model, the SEP values were 2.46, 0.92, and 1.04 g/L for fructose, acetate, and gluconacetan, respectively . Using only 14 standards, it was therefore possible to approach the performance of an 88-standard-based calibration model having SEP values of 1.11, 0.37, and 0.79 g/L for fructose, acetate, and gluconacetan, respectively . Therefore, the proposed algorithm is a valuable approach to reduce the calibration time of multiwavelength optical sensors.

Microb Pathog, 2004 Sep, 37(3), 155 - 61
Production of prostaglandin E2 in monocytes stimulated in vitro by Chlamydia trachomatis, Chlamydophila pneumoniae, and Mycoplasma fermentans; Krausse-Opatz B et al.; Chlamydia trachomatis (CT) as well as Chlamydophila pneumoniae (CP) cause chronic inflammatory diseases in humans . Persistently infected monocytes are involved in the pathogenesis by inducing mediators of inflammation . An in vitro system of chlamydial persistence in human peripheral blood monocytes (HPBM) was used to investigate prostaglandin E(2) (PGE(2)) production and the expression of the key enzyme for prostaglandin production, cyclooxygenase-2 (COX-2) . PGE(2) production was determined by PGE(2)-ELISA of HPBM-culture supernatants . Cox-2 mRNA expression was measured by real-time RT-PCR of total RNA isolated from HPBM . Both, CT and CP, stimulated PGE(2) production of HPBM in vitro . Equivalent numbers of CT per host cell induced a higher PGE(2)-response compared to CP . The amount of synthesized PGE(2) depended on the chlamydial multiplicity of infection (MOI) . Even at an MOI of 10 the amount of CT- and CP-induced prostaglandin, respectively, was lower than the amount of prostaglandin induced by E . coli lipopolysaccharide (LPS) at a concentration of 10microg/ml . In contrast to stimulation with LPS, Chlamydia-induced PGE(2) production as well as cox-2 mRNA decreased after day 1 post infection (p.i.) . These data indicate that Chlamydia stimulate PGE(2) production in human monocytes . Since Chlamydia are often contaminated by mycoplasma, the influence of mycoplasma on the prostaglandin production was investigated additionally . Mycoplasma fermentans (MF) also stimulated PGE(2) production . The co-infection of mycoplasma and Chlamydia resulted in an additive effect in the production of PGE(2) . Thus it is important to use host cells and Chlamydia free of mycoplasma contamination for the analysis of Chlamydia-induced prostaglandin production.

Food Chem Toxicol, 2004 Nov, 42(11), 1817 - 24
28-Day repeated dose toxicity study of dried microorganism in rats; Kitano M et al.; Ubidecarenone, also known as CoQ(10), is currently sold as a dietary supplement in the United States, with a majority of these products derived from the fermentation of carbohydrates or tobacco leaf extracts . In addition to its availability in dietary supplements, CoQ(10) is now being considered for use in foods . Accordingly, as part of the process for attaining "Generally Recognized as Safe" status, and to supplement information already available regarding the safety of CoQ(10) per se, a 28-day oral toxicity study in rats was conducted to evaluate the subacute safety of a microorganism biomass used as a new source in CoQ(10) production . Groups of Crj:CD(SD) rats (SPF) (6 males or females per group, 4 groups per sex) received dried microorganism at doses of 0, 500, 1000 or 2000 mg/kg/day via intragastric intubation . Clinical observations were recorded, and body weight, and food and water consumptions measured throughout the study . At the end of the study, aortic blood samples were collected from all animals for analysis of hematological and clinical chemistry parameters, and gross pathologic examination was performed . Histopathologic examination was performed on select tissues from the control and high-dose groups . There were no treatment-related changes that were considered to be of toxicological significance . Since rats treated with 2000 mg/kg of dried microorganism did not demonstrate any treatment-related changes, the no-observable-adverse-effect level (NOAEL) for dried microorganism was estimated to be greater than 2000 mg/kg/day under the present study conditions.

Appl Microbiol Biotechnol . 2004 Sep 3; {Epub ahead of print}
Improvement of pCOR plasmid copy number for pharmaceutical applications; Soubrier F et al.; Production of pharmaceutical-grade plasmid DNA is becoming important as the demand for clinical batches is steadily growing . pCOR plasmids have been specifically designed and used for gene delivery into humans, and have been produced by high cell-density fermentation with a yield of 100 mg/l . This yield could probably be increased as long as the release specifications of bulk plasmid remain the same, particularly in terms of plasmid sequence . We report here the use of genetic approaches in Escherichia coli to increase the copy number of pCOR . The bacterial gene encoding the pi initiator-protein, which plays a pivotal role in pCOR replication, was mutagenized . A fluorescence-based screening methodology in E . coli was used to identify novel copy-up mutations . A particular combination of copy-up mutations translated into a 3-5-fold increase in monomer pCOR plasmid DNA per biomass unit.

Microbiology, 2004 Sep, 150(Pt 9), 3065 - 74
Expression of Melanocarpus albomyces laccase in Trichoderma reesei and characterization of the purified enzyme; Kiiskinen LL et al.; Previous studies on Melanocarpus albomyces laccase have shown that this enzyme is very interesting for both basic research purposes and industrial applications . In order to obtain a reliable and efficient source for this laccase, it was produced in the filamentous fungus Trichoderma reesei . Two approaches were used: production of a non-fused laccase and a hydrophobin-laccase fusion protein . Both proteins were expressed in T . reesei under the cbh1 promoter, and significantly higher activities were obtained with the non-fused laccase in shake-flask cultures (corresponding to about 230 mg l(-1)) . Northern blot analyses showed rather similar mRNA levels from both expression constructs . Western analysis indicated intracellular accumulation and degradation of the hydrophobin-laccase fusion protein, showing that production of the fusion was limited at the post-transcriptional level . No induction of the unfolded protein response pathway by laccase production was detected in the transformants by Northern hybridization . The most promising transformant was grown in a fermenter in batch and fed-batch modes . The highest production level obtained in the fed-batch culture was 920 mg l(-1) . The recombinant laccase was purified from the culture supernatant after cleaving the major contaminating protein, cellobiohydrolase I, by papain . The recombinant and wild-type laccases were compared with regard to substrate kinetics, molecular mass, pH optimum, thermostability, and processing of the N- and C-termini, and they showed very similar properties.

Microbiology, 2004 Sep, 150(Pt 9), 2865 - 79
New roles for CDC25 in growth control, galactose regulation and cellular differentiation in Saccharomyces cerevisiae; Folch-Mallol JL et al.; Living organisms display large differences in stress resistance throughout their life cycles . To study the coordinated regulation of development and stress responses in exponentially growing yeast, mutants that displayed elevated heat-shock resistance at this stage were screened for . Here, two new mutant alleles of CDC25 in Saccharomyces cerevisiae, cdc25-21 and cdc25-22, are described . During exponential growth in glucose at 25 degrees C, these mutants are resistant to heat, oxidative, osmotic and ionic shock, accumulate stress-protein transcripts, show slow growth rates, thick cell walls and glycogen hyperaccumulation and lack cAMP signalling in response to glucose . Genetic and cellular analyses revealed that the stationary-phase phenotypes of cdc25-21 and cdc25-22 mutants are not due to entrance to a G(0) state during exponential growth, but are the result of a prolonged G(1) phase . It was found that, in the W303 background, CDC25 is dispensable for growth in glucose media . However, CDC25 is essential for growth in galactose, in non-fermentable carbon sources and under continuous incubation at 38 degrees C . In conclusion, the function of the catalytic, C-terminal domain of Cdc25p is not only important for fermentative growth, but also for growth in non-fermentable carbon sources and to trigger galactose derepression.

Appl Environ Microbiol, 2004 Sep, 70(9), 5323 - 30
Role of hexose transport in control of glycolytic flux in Saccharomyces cerevisiae; Elbing K et al.; The yeast Saccharomyces cerevisiae predominantly ferments glucose to ethanol at high external glucose concentrations, irrespective of the presence of oxygen . In contrast, at low external glucose concentrations and in the presence of oxygen, as in a glucose-limited chemostat, no ethanol is produced . The importance of the external glucose concentration suggests a central role for the affinity and maximal transport rates of yeast's glucose transporters in the control of ethanol production . Here we present a series of strains producing functional chimeras between the hexose transporters Hxt1 and Hxt7, each of which has distinct glucose transport characteristics . The strains display a range of decreasing glycolytic rates resulting in a proportional decrease in ethanol production . Using these strains, we show for the first time that at high glucose levels, the glucose uptake capacity of wild-type S . cerevisiae does not control glycolytic flux during exponential batch growth . In contrast, our chimeric Hxt transporters control the rate of glycolysis to a high degree . Strains whose glucose uptake is mediated by these chimeric transporters will undoubtedly provide a powerful tool with which to examine in detail the mechanism underlying the switch between fermentation and respiration in S . cerevisiae and will provide new tools for the control of industrial fermentations.

Arch Microbiol, 2004 Oct, 182(2-3), 204 - 11 Epub 2004 Aug 31.
Desulfosporomusa polytropa gen . nov., sp . nov., a novel sulfate-reducing bacterium from sediments of an oligotrophic lake; Sass H et al.; Five strains of sulfate-reducing bacteria were isolated from the highest positive dilutions of a most probable number (MPN) series supplemented with lactate and inoculated with sediments from the oligotrophic Lake Stechlin . The isolates were endospore-forming and were motile by means of laterally inserted flagella . They stained Gram-negative and contained b-type cytochromes . CO difference spectra indicated the presence of P582 as a sulfite reductase . Phylogenetic analyses of the 16S rDNA sequences revealed that the isolates were very closely affiliated with the genus Sporomusa . However, sulfate and amorphous Fe(OH)(3), but not sulfite, elemental sulfur, MnO(2), or nitrate were used as terminal electron acceptors . Homoacetogenic growth was found with H(2)/CO(2) gas mixture, formate, methanol, ethanol, and methoxylated aromatic compounds . The strains grew autotrophically with H(2) plus CO(2) in the presence or absence of sulfate . Formate, butyrate, several alcohols, organic acids, carbohydrates, some amino acids, choline, and betaine were also utilized as substrates . The growth yield with lactate and sulfate as substrate was 7.0 g dry mass/mol lactate and thus two times higher than in sulfate-free fermenting cultures . All isolates were able to grow in a temperature range of 4-37 degrees C . Physiologically and by the presence of a Gram-negative cell wall, the new isolates resemble known Desulfosporosinus species . However, phylogenetically they are affiliated with the Gram-negative genus Sporomusa belonging to the Selenomonas subgroup of the Firmicutes . Therefore, the new isolates reveal a new phylogenetic lineage of sulfate-reducing bacteria . A new genus and species, Desulfosporomusa polytropa gen . nov., sp . nov . is proposed.

Eur J Nutr . 2004 Sep 6; {Epub ahead of print}
Stimulatory effect of inulin on intestinal absorption of calcium and magnesium in rats is modulated by dietary calcium intakesShort- and long-term balance studies; Coudray C et al.; Previous studies have shown that short-term intake of fermentable oligosaccharides (OS), including inulin, can increase mineral intestinal absorption in humans and animals . While the stimulatory effect of these substances on intestinal magnesium (Mg) absorption is generally high and consistent, their effect on calcium (Ca) seems to depend on experimental conditions, particularly the duration of fermentable OS intake . The aim of this study was to determine how the short- and long-term dietary Ca intake may modulate the effect of inulin on Ca absorption . Sixty male Wistar rats, weighing 275 g, were randomized into two groups to receive or not 10% of inulin in their diet . Each group was divided into three sub-groups to receive one of the following dietary Ca levels 0.25%, 0.50% and 0.75% in their food . The animals were fed fresh food and water ad libitum for 40 days . Apparent intestinal absorptions of Ca and Mg were determined at D13 and D36 of the experiment . As expected, inulin feeding increased Ca and Mg absorption in both periods at all dietary Ca levels . However, the effect of inulin on intestinal Ca absorption was dependent on dietary Ca levels and on experiment duration . In the short-term period, the inulin effect was prominent in the groups receiving high or low Ca levels, but in long-term period inulin improved intestinal Ca absorption much more in the group receiving the low Ca level . In addition, efficiency of intestinal absorption of Ca and Mg (%) was negatively affected by Ca intake levels . These results show that the beneficial effect of inulin on intestinal Ca absorption may be more marked in cases where the Ca intake is low or where the organism's Ca requirement is high . Further studies are required to confirm these results in humans.

Zhong Xi Yi Jie He Xue Bao, 2004 May, 2(3), 216 - 8
{A preliminary report on solid-state fermentation of Ganoderma lucidum with Radix Astragali containing medium}; Chen HZ et al.; OBJECTIVE: To test the practicability of the solid-state fermentation for medicinal fungi by fermenting Ganoderma lucidum with Radix Astragali containing medium . METHODS: Ganoderma lucidum was fermented in ordinary medium, drug-containing medium (containing Radix Astragali) and selenium-rich drug-containing medium respectively . The polysaccharide contents of fermentation products from the three kinds of culture media were tested at different time, and the changes were compared . RESULTS: The polysaccharide contents of fermentation products from the three kinds of culture media were 4.65%, 3.76% and 4.50% respectively and their relative standard deviation were 1.61%, 1.99% and 1.86% respectively . By observing the changes of the contents of polysaccharide, protein and total saponin in fermentation products from the drug-containing medium at different time, it was found that the 28th fermentation day was the time when secondary metabolism was most active, and it should be the fermented terminal point . CONCLUSION: The fermentative combination of Ganoderma lucidum and Radix Astragali is practicable.

Appl Microbiol Biotechnol . 2004 Aug 25; {Epub ahead of print}
Production of bacterial cellulose by Acetobacter xylinum BPR2001 using molasses medium in a jar fermentor; Bae SO et al.; Bacterial cellulose (BC) production by Acetobacter xylinum subsp . sucrofermentans BPR2001 using molasses medium was carried out in a jar fermentor . When molasses was subjected to H(2)SO(4)-heat treatment, the maximum BC concentration increased to 76% more than that achieved using untreated molasses, and the specific growth rate increased 2-fold . When the initial sugar concentrations in the H(2)SO(4)-heat treated molasses were varied from 23 g/l to 72 g/l, BC concentration, production rate, and yield were maximum at sugar concentrations of 23 g/l and 37 g/l, and production of by-products, such as polysaccharides and CO(2), was lower than at sugar concentrations of 48 g/l and 72 g/l, indicating that maintaining a lower molasses concentration is essential for efficient BC production in jar fermentors, this being due mainly to the complex nature of molasses . Molasses has a clear advantage over pure sugars as a carbon source from an economic viewpoint.

J Pharm Biomed Anal, 2004 Sep 3, 35(5), 1101 - 12
Identification and chemical profiling of monacolins in red yeast rice using high-performance liquid chromatography with photodiode array detector and mass spectrometry; Li YG et al.; Monascus purpureus-fermented rice (red yeast rice) was one of the food supplements that had the ability of lowering the blood-lipid levels, and monacolins have been proved to be main active constituents . In total 14 monacolin compounds such as monacolin K (mevinolin), J, L, M, X, and their hydroxy acid form, as well as dehydromonacolin K, dihydromonacolin L, compactin, 3alpha-hydroxy-3,5-dihydromonacolin L, etc . were identified in red yeast rice, using high-performance liquid chromatography with photodiode array detector and tandem mass spectrometry . A chemical fingerprint profiling method to display bioactive monacolins in red yeast rice was established and could be used for the quality control of the target material and its related products . Ten finish products labeled as red yeast rice from different manufacturers in marketing were traced using the chromatographic chemical profiling method, and the results show that only two of them were similar while the other eight were significantly different from the reference red yeast rice . All of these materials including raw material powder and finished products available were quantified and the contents of monacolins were calculated with reference of monacolin K (mevinolin) as the standard.

J Nat Prod, 2004 Aug, 67(8), 1396 - 9
Dictyosphaeric acids A and B: new decalactones from an undescribed Penicillium sp . obtained from the alga Dictyosphaeria versluyii; Bugni TS et al.; Fungal isolate F01V25 was obtained from the alga Dictyosphaeria versluyii collected near Dravuni, Fiji, in 2001 and represented a previously undescribed Penicillium sp . Fermentation of isolate F01V25 resulted in the production of two new polyketides, dictyosphaeric acids A and B, along with the known anthraquinone carviolin . The relative stereochemistry of dictyosphaeric acids A and B was determined using the J-based configuration analysis method in conjunction with ROE and NOE correlations.

East Mediterr Health J, 2001 Jul-Sep, 7(4-5), 756 - 62
Detection and characterization of verotoxin-producing strains among sorbitol non-fermenting Escherichia coli; Jafari A et al.; The presence of genes for verotoxin 1 and 2 (VT1 and 2) among sorbitol non-fermenting Escherichia coli isolates from diarrhoeal cases was assessed using polymerase chain reaction assay . Of 60 (88%) positive isolates, 37 (62%) harboured VT1 and 23 (38%) both VT1 and VT2 . In HeLa cell adherence assay, 48 (71%) isolates exhibited mannose-resistant adherence to HeLa cells . Multidrug resistance was observed in 56 (82%) isolates, with ampicillin, chloramphenicol, streptomycin, sulfamethoxazole-trimethoprim and tetracycline pattern being the most common . There were 13 common and 22 single biochemical phenotypes identified . Isolates belonging to common biochemical phenotypes normally had a similar pattern of adherence and VT production, but differed greatly in their pattern of antibiotic resistance, pointing to a high rate of antibiotic-resistance transfer among these isolates.

Arch Latinoam Nutr, 2004 Mar, 54(1), 36 - 44
{Effects of black bean Phaseolus vulgaris consumption on the nutritional status of Guatemalan population}; Serrano J et al.; Guatemala provides an example of epidemiological superposition, in which health problems typical of developed countries and developing countries are both observed . Nutritional deficiencies in some micronutrients like vitamin A and iron coexist alongside chronic diseases such as diabetes type II and cardiovascular diseases . The importance of black beans in the normal Guatemala diet is well known:70g per capita of black beans are consumed daily . Black beans are an important sources of protein and energy in the diet . They contain "lente" digestion carbohydrates and a high proportion of non-digested carbohydrates that may be fermented in the large intestine . Theses types of carbohydrates are associated with a low glycemic response, low serum cholesterol levels, and a decrease of colon cancer risk factors . These physiological effects may be related to colonic fermentation end products (propionic and butyric acids) . Black beans also contain several antinutritional compounds (enzymatic inhibitors, haemaglutenins, saponins and phytic acid, etc.), some of them thermolabiles that are partially eliminated during culinary processes and may modify the nutritional quality of beans . Black beans play a crucial role in the etiology of several diseases in Guatemala.

Blood, 2004 Dec 15, 104(13), 4252 - 9 Epub 2004 Dec 15.
Mycoplasma fermentans infection promotes immortalization of human peripheral blood mononuclear cells in culture; Zhang S et al.; Chronic infection or colonization by mycoplasma(s) could gradually and significantly alter many biologic properties of mammalian host cells in culture, including induction of malignant transformation . We examined effects of Mycoplasma fermentans infection on the continuing survival and immortality of human peripheral blood mononuclear cells (PBMCs) from healthy blood donors . Without specific supplemental growth factors, human PBMCs normally die rapidly, with few cells other than macrophages/monocytes surviving after 2 weeks in cultures . Only occasional Epstein-Barr virus (EBV)-positive B lymphocytes would continue to proliferate and undergo spontaneous immortalization . Our present study revealed that infection of human PBMCs in culture with the incognitus and PG18 strains of M fermentans, but surprisingly not with some other strains tested in parallel, markedly enhanced the rate of EBV-positive B lymphocytes to undergo immortalization (74% vs 17%) . Compared with spontaneously immortalized PBMCs, the PBMCs immortalized in cultures infected with the mycoplasmas often had prominent karyotype changes with chromosomal loss, gain, or translocations . Furthermore, many of these immortalized B lymphocytes were found to be monoclonal in nature . The in vitro findings would be of relevance to lymphoproliferative disorders that occurred in patients with immune suppression . The mycoplasma-mediated promotional effect in cell immortalization and its potential clinical implications warrant further study.

Trends Biotechnol, 2004 Sep, 22(9), 477 - 85
Production of bioenergy and biochemicals from industrial and agricultural wastewater; Angenent LT et al.; The building of a sustainable society will require reduction of dependency on fossil fuels and lowering of the amount of pollution that is generated . Wastewater treatment is an area in which these two goals can be addressed simultaneously . As a result, there has been a paradigm shift recently, from disposing of waste to using it . There are several biological processing strategies that produce bioenergy or biochemicals while treating industrial and agricultural wastewater, including methanogenic anaerobic digestion, biological hydrogen production, microbial fuel cells and fermentation for production of valuable products . However, there are also scientific and technical barriers to the implementation of these strategies.

J Dairy Sci, 2004 Aug, 87(8), 2578 - 90
Grain processing, forage-to-concentrate ratio, and forage length effects on ruminal nitrogen degradation and flows of amino acids to the duodenum; Yang WZ et al.; The objectives of this study were to evaluate effects of dietary factors that alter ruminal fermentability on rumen N degradation, microbial protein synthesis, duodenal flows, and digestibility of amino acids (AA) in the intestines and the total tract . The experiment was a double 4 x 4 quasi-Latin square with a 2(3) factorial arrangement of treatments . The dietary factors were extent of barley grain processing, coarse (processing index; PI = 75.5%) or flat (PI = 60.2%); forage-to-concentrate (F:C) ratio, low (35:65) or high (55:45) on a DM basis; and forage particle length (FPL), long (7.59 mm) or short (6.08 mm) . Eight lactating cows with ruminal and duodenal cannulas were offered ad libitum access to a total mixed diet . There were no significant interactions between dietary treatments for ruminal N degradation or its duodenal flow and digestibility in the intestines . Passage of microbial protein to the duodenum was improved with increased F:C ratio of the diet but was not affected by grain processing or FPL . Ruminal digestibility of N was increased with increased F:C ratio (49 vs . 60%) and with reduced FPL (59 vs . 50%) . Increased grain processing improved N digestibility both in the intestine (15%) and in the total tract (8%) . Reduction in the FPL of the diets reduced intestinal N digestion by 14% without affecting the N digestion in the total tract . Increased extent of grain processing tended to enhance duodenal flows of AA . In contrast, reducing FPL lowered flows of dietary AA to the duodenum because of lowered flows of feed plus endogenous N . Increased F:C ratio of the diet did not change the flow of total AA, but there was a reduced flow of dietary AA and increased flow of microbial AA . Flows of several individual AA were increased by feeding flatly rolled barley with limited effects of F:C ratio or FPL . An interaction between grain processing and FPL was detected for flows of some AA . Diets formulated with flatly rolled barley plus long FPL increased Arg, Thr, Asp, Glu, Ser, Tyr, and nonessential AA (NEAA) by more than 24%, compared with other combinations of grain processing and FPL . Digestibility of essential AA (EAA) in the intestine (68%) was higher than that of NEAA (63%), but digestion of total AA (65%) was similar to that of total N (66%) . Digestibilities of individual AA in the intestine ranged from 46 to 77% and were generally improved with increased grain processing . However, effects of F:C ratio or FPL on digestion of AA were limited . These results indicate that manipulation of dairy cow diets can improve ruminal N degradation, microbial protein synthesis, flows of AA to the duodenum, and intestinal digestibility of AA . Combining dietary factors can be more beneficial than changing individual dietary factors for improving the delivery of AA to the small intestine.

J Dairy Sci, 2004 Aug, 87(8), 2495 - 505
Replacing chopped alfalfa hay with alfalfa silage in barley grain and alfalfa-based total mixed rations for lactating dairy cows; Plaizier JC; The effects of replacing chopped alfalfa hay with alfalfa silage in a fine barley grain and alfalfa-based total mixed ration (TMR) were evaluated . Diets contained (dry matter basis) 53.0% commercial energy supplement, 10.3% commercial protein supplement, and 9.7% corn silage . Diets varied in inclusion of chopped alfalfa hay and alfalfa silage, and contained either 20.0% chopped alfalfa hay and 7.0% alfalfa silage, 10.0% chopped alfalfa hay and 17.0% alfalfa silage, or 27.0% alfalfa silage . Contents of crude protein, neutral detergent fiber (NDF), acid detergent fiber, and minerals did not differ among diets . Replacing chopped alfalfa hay with alfalfa silage decreased dietary dry matter, and increased dietary soluble protein and physical effective NDF calculated as the proportion of dietary NDF retained by the 8- and 19-mm screens of the Penn State Particle Separator (peNDF(NDF)) from 13.3 to 15.6% DM . Replacing chopped alfalfa hay with alfalfa silage did not affect dry matter intake, rumen pH, rumen volatile fatty acids, blood lactate, milk fat, and milk protein percentage, but did decrease blood glucose, tended to increase blood urea, and numerically decreased milk yield and milk protein yield . A wider range in peNDF(NDF) and a higher inclusion of corn silage might have resulted in greater differences in rumen fermentation and milk production among diets . The pH of rumen fluid samples collected 4 h after feeding varied from 5.90 to 5.98, and milk fat percentage varied from 2.50 to 2.60% among diets . These values suggest that mild subacute ruminal acidosis was induced by all diets.

J Dairy Sci, 2004 Aug, 87(8), 2486 - 94
Feeding lactose increases ruminal butyrate and plasma beta-hydroxybutyrate in lactating dairy cows; DeFrain JM et al.; Ruminal fermentation of lactose increases molar proportions of butyrate, which is metabolized by the ruminal epithelium to beta-hydroxybutyrate (BHBA) . To determine the effects of dietary whey, and specifically lactose, on concentrations of ruminal and blood volatile fatty acids (VFA) and blood BHBA, 8 Holstein and 4 Brown Swiss multiparous cows (210 +/- 33 d in milk) were blocked by breed and randomly assigned to one of three 4 x 4 Latin squares . Treatments were control (CON; 7.1% of dietary dry matter {DM} as cornstarch), liquid whey (WHEY; 9.4% of diet DM) containing 70% lactose on a DM basis, low lactose (LOLAC; 7.1% lactose), or high lactose (HILAC; 14.3% lactose) . Diets contained 53% forage as corn silage, alfalfa hay, and grass hay (DM basis) and a corn and soybean meal-based concentrate . Average dietary percentage of crude protein and energy density (Mcal/kg net energy for lactation) were 16.8 and 1.47, respectively . Feeding lactose increased DM intake . Milk production and composition were not affected by diet with the exception of decreased urea nitrogen in milk from cows fed lactose . Greater proportions of ruminal propionate were observed in cows fed CON relative to those fed WHEY and LOLAC . Increasing dietary lactose increased proportions of ruminal butyrate and decreased acetate and branched-chain VFA . Concurrent with the increase in ruminal butyrate concentrations, there was an increase in plasma BHBA as lactose in the diet increased . Concentrations of VFA in plasma were not affected by diet with the exception of the branched-chain VFA, which were increased in cows fed LOLAC compared with WHEY . These data indicate lactose fermentation increases proportions of ruminal butyrate and plasma BHBA in lactating dairy cows; however, the observed increase in plasma BHBA is not sufficient to subject cows to ketosis.

Proc Natl Acad Sci U S A, 2004 Sep 7, 101(36), 13318 - 23 Epub 2004 Aug 23.
Identification of a quinone-sensitive redox switch in the ArcB sensor kinase; Malpica R et al.; Escherichia coli senses and signals anoxic or low redox conditions in its growth environment by the Arc two-component system . Under anaerobic conditions, the ArcB sensor kinase autophosphorylates and transphosphorylates ArcA, a global transcriptional regulator that controls the expression of numerous operons involved in respiratory or fermentative metabolism . Under aerobic conditions, the kinase activity of ArcB is inhibited by the quinone electron carriers that act as direct negative signals . Here, we show that the molecular mechanism of kinase silencing involves the oxidation of two cytosol-located redox-active cysteine residues that participate in intermolecular disulfide bond formation, a reaction in which the quinones provide the source of oxidative power . Thus, a pivotal link in the Arc signal transduction pathway connecting the redox state of the quinone pool to the transcriptional apparatus is elucidated.

J Antibiot (Tokyo), 2004 Jun, 57(6), 373 - 8
Mutactimycin PR, a new anthracycline antibiotic from Saccharothrix sp . SA 103 . II . Physico-chemical properties and structure elucidation; Zitouni A et al.; A new antibiotic termed mutactimycin PR (1) was isolated along with the known mutactimycin C (2) from the fermentation broth of Saccharothrix sp . SA 103 . The two compounds belong to the anthracycline group . The structure of these antibiotics was elucidated with the aid of NMR and mass spectrometric investigations . The novel compound mutactimycin PR was characterized as 5,12 Naphtacenedione, 7-{(6-deoxy-3-O-methyl-alpha-L-mannopyranosyl)oxy}-4-{(6-deoxy-alpha-L-mannopyranosyl)oxy}-7,8,9,10-tetrahydro-6,9,11-trihydroxy-9 methyl.

Infect Immun, 2004 Sep, 72(9), 5004 - 11
Mycoplasma fermentans binds to and invades HeLa cells: involvement of plasminogen and urokinase; Yavlovich A et al.; Adherence of Mycoplasma fermentans to HeLa cells followed saturation kinetics, required a divalent cation, and was enhanced by preincubation of the organism at 37 degrees C for 1 h in a low-osmolarity solution . Proteolytic digestion, choline phosphate, or anti-choline phosphate antibodies partially inhibited the adherence, supporting the notion that M . fermentans utilizes at least two surface components for adhesion, a protease-sensitive surface protein and a phosphocholine-containing glycolipid . Plasminogen binding to M . fermentans greatly increased the maximal adherence of the organism to HeLa cells . Anti-plasminogen antibodies and free plasminogen inhibited this increase . These observations suggest that in the presence of plasminogen the organism adheres to novel sites on the HeLa cell surface, which are apparently plasminogen receptors . Plasminogen-bound M . fermentans was detected exclusively on the cell surface of the infected HeLa cells . Nevertheless, plasminogen binding in the presence of the urokinase-type plasminogen activator (uPA) promoted the invasion of HeLa cells by M . fermentans . The latter finding indicates that the invasiveness of M . fermentans does not result from binding plasminogen but from activation of the bound plasminogen to plasmin . Cholesterol depletion and sequestration with beta-cyclodextrin and filipin, respectively, did not affect the capacity of M . fermentans to adhere, but invasion of HeLa cells by uPA-activated plasminogen-bound M . fermentans was impaired, suggesting that lipid rafts are implicated in M . fermentans entry.

FEMS Microbiol Lett, 2004 Aug 15, 237(2), 425 - 30
FLO11 is essential for flor formation caused by the C-terminal deletion of NRG1 in Saccharomyces cerevisiae; Ishigami M et al.; The flor strains of Saccharomyces cerevisiae form a flor on the surface of wine after alcoholic fermentation . High hydrophobicity of the cell surface is suggested to be important for flor formation by the flor wine yeasts . However, the molecular mechanism of flor formation is not clear . We found that expression of C-terminal deleted NRG1 lacking its two C2H2 zinc finger motifs (NRG1(1-470)) on the multicopy plasmid conferred the ability to form a flor to a non-flor laboratory strain . The cell surface hydrophobicity of NRG1(1-470) was higher than of the non-flor strain . Disruption of the Nrg1p-repressed gene FLO11, which encodes a cell surface glycoprotein that functions as a flocculin or an adhesin, abolished flor formation . Moreover, expression of FLO11 on a multicopy plasmid could also cause flor formation . These results indicate that FLO11 is essential for flor formation by NRG1(1-470) . In addition, the results suggest that the C-terminal truncated form of Nrg1p exerts a dominant negative effect on FLO11 repression, resulting in FLO11 expression and, thus, flor formation.

Ying Yong Sheng Tai Xue Bao, 2004 May, 15(5), 863 - 6
{Production and properties of chitinase from Beauveria bassiana Bb174 in solid state fermentation}; Zhang J et al.; This paper studied the chitinase production of Beauveria bassiana Bb174 under solid state fermentation condition . The optimal medium consisted of wheat bran and silkworm chrysalis at the ratio of 4:1, supplemented with 1 g peptone L(-1) as nitrogen source and some other mineral nutrients . The enzyme activity reached 126 units per gram dry medium after cultured for 2 days at 28 degrees C and natural pH by inoculated 3 ml spore suspension into this medium . The optimal temperature and pH for chintinase production were 40 degrees C and 5.0, respectively . The temperature to lose 50% activity of the enzyme was 48 degrees C after incubated at 30-70 degrees C for 1 h . The enzyme was stable at 30-40 degrees C and pH 4-6, and the Km and Vmax values were 0.52 mg x ml(-1) and 0.7 deltaE680 x h(-1), respectively.

J Anim Sci, 2004 Aug, 82(8), 2384 - 91
Influence of concentrate composition and forage type on retail packaged beef quality; O'Sullivan A et al.; The objective of this study was to determine the effect of type of conserved forage and concentrate composition on the quality of beef held in overwrapped (aerobic) or modified atmosphere packaging under simulated retail display for 17 d . Friesian steers (n = 45) were assigned randomly to one of five dietary treatments: 1) extensively fermented grass silage plus silage concentrate (EFS); 2) restricted fermented grass silage plus silage concentrate (RFS); 3) starch-based concentrate plus wheat straw (SC); 4) nonstarch-based concentrate plus wheat straw (NSC); or 5) zero-grazed perennial ryegrass plus grass concentrate (RYE) . Meat quality was determined by measuring color, lipid oxidation (TBARS), alpha-tocopherol concentrations, and fatty acid composition . In aerobically packaged beef, there was a display x diet interactive effect (P < 0.001) on Hunter a* values, with steaks from the EFS group having higher (P < 0.05) a* values than all other dietary groups from d 6 through d 17 . Moreover, during the last 12 d of display, beef from the EFS group had the lowest (P < 0.01) proportion of metmyoglobin (display day x diet; P < 0.001) . Under aerobic packaging, the SC and NSC groups produced steaks with higher (P < 0.05) TBARS values than RFS, EFS, and RYE groups, which did not differ from each other (display day x diet; P < 0.01) . The SC and NSC groups had higher (P < 0.05) oxidation levels than RFS, EFS, and RYE groups, which did not differ from each other . Beef from the EFS group had (P < 0.05) higher concentrations of alpha-tocopherol than from the SC, NSC, and RYE groups . Beef from EFS-fed steers had a higher (P < 0.05) proportion of saturated fatty acids than the SC and NSC groups . It was concluded that the method of grass conservation influenced beef color, whereas concentrate composition did not . Color of aerobically packaged beef was improved by feeding animals silage that had undergone extensive fermentation . Conversely, oxidative stability was decreased by feeding animals starch- and nonstarch-based concentrate diets.

J Chromatogr A, 2004 Jul 16, 1043(1), 3 - 7
Potential of fermentation profiling via rapid measurement of amino acid metabolism by liquid chromatography-tandem mass spectrometry; Dalluge JJ et al.; Monitoring amino acid metabolism during fermentation has significant potential from the standpoint of strain selection, optimizing growth and production in host strains, and profiling microbial metabolism and growth state . A method has been developed based on rapid quantification of underivatized amino acids using liquid chromatography-electrospray tandem mass spectrometry (LC-MS-MS) to monitor the metabolism of 20 amino acids during microbial fermentation . The use of a teicoplanin-based chiral stationary phase coupled with electrospray tandem mass spectrometry allows complete amino acid analyses in less than 4 min . Quantification is accomplished using five isotopically labeled amino acids as internal standards . Because comprehensive chromatographic separation and derivatization are not required, analysis time is significantly less than traditional reversed- or normal-phase LC-based amino acid assays . Intra-sample precisions for amino acid measurements in fermentation supernatants using this method average 4.9% (R.S.D.) . Inter-day (inter-fermentation) precisions for individual amino acid measurements range from 4.2 to 129% (R.S.D.) . Calibration curves are linear over the range 0-300 microg/ml, and detection limits are estimated at 50-450 ng/ml . Data visualization techniques for constructing semi-quantitative fermentation profiles of nitrogen source utilization have also been developed and implemented, and demonstrate that amino acid profiles generally correlate with observed growth profiles . Further, cellular growth events, such as lag-time and cell lysis can be detected using this methodology . Correlation coefficients for the time profiles of each amino acid measured illustrate that while several amino acids are differentially metabolized in similar fermentations, a select group of amino acids display strong correlations in these samples, indicating a sub-population of analytes that may be most useful for fermentation profiling.

J Agric Food Chem, 2004 Aug 25, 52(17), 5396 - 403
Determination of piceid and resveratrol in Spanish wines deriving from Monastrell (Vitis vinifera L.) grape variety; Moreno-Labanda JF et al.; The presence of stilbenes in wine is becoming an important issue due to their claimed relation to a low incidence in coronary diseases and their increasing implication as cancer chemopreventive and neuroprotective agents . Total resveratrol content, quantified as glucoside and aglycone forms of resveratrol, has been determined in a survey of 45 Monastrell monovarietal Spanish red wine types (around 135 wine samples), belonging to Alicante and Bullas appellations . The average between ratio glucoside/aglycone forms of resveratrol in these wines was considerably high, ranging from 82 to 91% of resveratrol in its glycosidic form . This characteristic was observed in a high percentage of the studied wines, which were made under different winemaking procedures, and from different vintages (1995-2002) . In addition, wines made using macerative fermentations with double amount of solid parts ("doble pasta") reached the highest levels of total stilbene content expressed as resveratrol equivalent, i.e., 30 mg/L (average of 18.8 mg/L) . It can be concluded that high resveratrol glucoside concentration and low free isomer content can be considered characteristics of the Monastrell variety, as it happens to red wines deriving from other varieties grown at warm climates . This fact, also observed for other French and Portuguese red varieties, might play an important role in food habits involving these types of wines.

Indian J Environ Health, 2003 Jul, 45(3), 231 - 4
Feasibility of some treatments for improving the composting of municipal solid waste; Babyranidevi S et al.; During composting limited degradation of organic matter occurs due to the influence of various factors such as presence of lignin, resistant nature of cellulose, high temperature, etc . During windrow composting, temperature within the waste mass rises and limitation in degradability of organic matter, studies on composting in temperature range of 45-60 degrees C were carried out . With a view to observe the effect of pretreatment for improving composting, Municipal Solid Waste (MSW) was subjected to alkali treatment using lime, and ammonia gas circulation in the waste mass . In practice aerobic condition within the windrow is ensured by turning/mixing the waste 5-6 times during the composting period . In the present study to find out the extent of aerobic condition, the waste mass was composted in the masony brick tank which was provided with honeycomb opening from all the sides . Similarly composting was also carried out on perforated false bottom . Various parameters such as fermentable organic substance (FOS), carbon to nitrogen (C/N) ratio and temperature were monitored for the waste mass undergoing composting . The study reveals that experiments with aeration from bottom as well as surface followed by alkaline treatment give better composting performance.

J Biotechnol, 2004 Sep 9, 112(3), 279 - 88
Comparison of the Sec and Tat secretion pathways for heterologous protein production by Streptomyces lividans; Schaerlaekens K et al.; Streptomyces is an interesting host for the secretory production of recombinant proteins because of its natural ability to secrete high levels of active proteins into the culture broth and the availability of extensive fermentation knowledge . In bacterial expression systems, heterologous protein secretion has, so far, almost exclusively been investigated using signal peptides that direct the secretion to the Sec pathway . In this study, we assessed the possibility of the Streptomyces lividans twin-arginine translocation (Tat) pathway to secrete the human proteins tumor necrosis factor (TNF) alpha and interleukin (IL) 10 by fusing the coding sequences of mature hTNFalpha and hIL10 to the twin-arginine signal peptides of S . lividans xylanase C (XlnC) and Streptomyces antibioticus tyrosinase . Both proteins were secreted and this secretion was blocked in the DeltatatB and DeltatatC single mutants, indicating that the transport of hTNFalpha and hIL10 could be directed through the Tat pathway . Secretion levels of hTNFalpha and hIL10, however, were lower for Tat-dependent than for Sec-dependent transport using the Sec-dependent signal peptide of the Streptomyces venezuelae subtilisin inhibitor . Surprisingly, Sec-dependent transport was enhanced in the tatB deletion strain . This was especially interesting in the case of hIL10, where Sec-dependent transport of hIL10 was at least 15 times higher in the DeltatatB mutant than in the wild-type strain.

Immunology, 2004 Sep, 113(1), 121 - 9
Lipid-associated membrane proteins of Mycoplasma fermentans and M . penetrans activate human immunodeficiency virus long-terminal repeats through Toll-like receptors; Shimizu T et al.; Mycoplasmas are known to enhance human immunodeficiency virus (HIV) replication, and mycoplasma-derived lipid extracts have been reported to activate nuclear factor-kappaB (NF-kappaB) through Toll-like receptors (TLRs) . In this study, we examined the involvement of TLRs in the activation of HIV long-terminal repeats (LTR) by mycoplasma and their active components responsible for the TLR activation . Lipid-associated membrane proteins (LAMPs) from two species of mycoplasma (Mycoplasma fermentans and M . penetrans) that are associated with acquired immune-deficiency syndrome (AIDS), were found to activate HIV LTRs in a human monocytic cell line, THP-1 . NF-kappaB deletion from the LTR resulted in inhibition of the activation . The LTR activation by M . fermentans LAMPs was inhibited by a dominant negative (DN) construct of TLR1 and TLR6, whereas HIV LTR activation by M . penetrans LAMPs was inhibited by DN TLR1, but not by DN TLR6 . These results indicate that the activation of HIV LTRs by M . fermentans and M . penetrans LAMPs is dependent on NF-kappaB, and that the activation of HIV LTR by M . fermentans LAMPs is mediated through TLR1, TLR2 and TLR6 . In contrast, the LTR activation by M . penetrans LAMPs is carried out through TLR1 and TLR2, but not TLR6 . Subsequently, the active component of M . penetrans and M . fermentans LAMPs was purified by reverse-phase high-performance liquid chromatography (HPLC) . Interestingly, the purified lipoprotein of M . penetrans LAMPs (LPMp) was able to activate NF-kappaB through TLR1 and TLR2 . On the other hand, the activation of NF-kappaB by purified lipoprotein of M . fermentans LAMPs (LPMf) was mediated through TLR2 and TLR6, but not TLR1.

Med Secoli, 2003, 15(2), 269 - 90
{Fermentation as the origin of life: discussions on blood in Italy in the late 17th century}; Conforti M; The article examines the correspondence (1701) between the Neapolitan mathematician Giacinto De Cristofaro and Domennico Guglielmini, professor of theoretical medicine at Padua, on the role of blood and on the fermentative process in the 'origin' of life . The discussion is set against the background of the lively Italian medical debates and experimentations on the function and composition of the blood . Works by Marcello Malpighi, Giacomo Sandri, Giovanni Maria Lancisi, Giorgio Baglivi, Giovanbattista Morgagni and Nicola Cirillo - all of them taking into account Robert Boyle's Natural History of the Blood (1683-4) - show that blood composition and heart motion were described in different ways, ranging from the adoption of chemical theories and experimentation to that of strictly mechanical explanations . Different positions about the role of the blood and of its fermentative motion reflect different views about the relationship between matter and life.

Med Secoli, 2003, 15(2), 247 - 67
{Substance and matter in an unpublished work by Girolamo Brasavola}; Favino F; The article examines an unpublished work by Girolamo Brasavola, a physician from ferrara who lived and worked in Rome in the second half of the 17th century . This is the chapter De fermentatione of the lost Institutions Philosophico -Medicae by the same author . The work somehow reached Cardianl Gacomo Nini before 1680 and has been afterwards kept among the manuscripts of the Vatican Library . The article examines the materialistic and corpuscular philosophy and the medical theory adopted by Brasavola identifying their sources-Sennert, Gassendi Willis, Descartes-and illustrating their philosophical background.

Med Secoli, 2003, 15(2), 227 - 45
{Chemistry of life: ferments and fermentation in 17th-century iatrochemistry}; Clericuzio A; The concepts of ferment and fermentation played an important, though heretofore neglected, role in 17th-century physiology . Though these notions can be found in ancient philosophy and medicine, as well as in medieval medicine, they became integral part of the chemical medicine that was advocated by Paracelsus and his school . Paracelsians made fermentation a central concept in their successful effort to give chemical foundation to medicine . Jean Baptiste van Helmont and Sylvius used the concepts of ferment and fermentation to explain a variety of physiological processes in human body . Corpuscular philosophers like Robert Boyle and Thomas Willis reinterpreted these notions in corpuscular terms and separated the concept of ferment from that of fermentation . In the second half of the seventeenth century, physiologist tried to explain fermentation by means of chemical reactions, as for instance acid -alkali, and ruled out the notion of ferment as superfluous to their investigations . At the end of hte seventeenth century fermentation attracted the interest of physicists like Johannes Bernoulli and Isaac Newton, who tried to explain fermentative processes in terms of matter and motion (Bernoulli) and short-range forces (Newton) . George Ernst Stahl devoted a work to fermentation: the Zymotechnia . He explained fermentation as the outcome of the reactions of molecules formed of saline, oily and earthy corpuscles with particles of water . He saw fermentation as a mechanical process, i.e . as collision of different kinds of corpuscles.

J Plant Physiol, 2004 Jul, 161(7), 803 - 8
Sugar utilization and anoxia tolerance in rice roots acclimated by hypoxic pretreatment; Kato-Noguchi H; Although most cereal roots cannot elongate under anoxic conditions, primary roots of three-day-old rice (Oryza sativa L.) seedlings were able to elongate during a 24-h period of anoxia . Hypoxic pretreatment (H-PT) increased the elongation of their roots . Sucrose synthase (EC 2.4.1.13), glucokinase (EC 2.7.1.2), fructokinase (EC 2.7.1.4), pyruvate decarboxylase (EC 4.1.1.1) and alcohol dehydrogenase (EC 1.1.1.1) activities were increased by anoxia in both H-PT and non-pretreated (N-PT) roots . However, these activities were greater in the H-PT roots than in the N-PT roots . The average rate of production of ethanol for the initial 6h after the onset of anoxia was 3.7 and 1.4 micromolg(-1) fresh weight h(-1) for the H-PT and N-PT roots, respectively, suggesting that ethanolic fermentation may increase more quickly in the H-PT roots than in the N-PT roots . Roots of the seedlings lost ATP and total adenine nucleotides in anoxia, however, the H-PT roots maintained higher levels of ATP and total adenine nucleotides compared to the N-PT roots . These results show that rice roots are able to utilize the set of enzymes involved in the metabolism of soluble sugars under anoxia . The ability to maintain an active fermentative metabolism for production of ATP by fueling the glycolytic pathway with fermentable carbohydrate is probably greater in H-PT than in N-PT roots.

J Anim Sci, 2004 Jul, 82(7), 2185 - 92
Effect of field pea level on intake, digestion, microbial efficiency, ruminal fermentation, and in situ disappearance in beef steers fed forage-based diets; Reed JJ et al.; Four ruminally and duodenally cannulated crossbred beef steers (397+/-55 kg initial BW) were used in a 4 x 4 Latin square to evaluate the effects of increasing level of field pea supplementation on intake, digestion, microbial efficiency, ruminal fermentation, and in situ disappearance in steers fed moderate-quality (8.0% CP, DM basis) grass hay . Basal diets, offered ad libitum twice daily, consisted of chopped (15.2-cm screen) grass hay . Supplements were 0, 0.81, 1.62, and 2.43 kg (DM basis) per steer daily of rolled field pea (23.4% CP, DM basis) offered in equal proportions twice daily . Steers were adapted to diets on d 1 to 9; on d 10 to 14, DMI were measured . Field pea and grass hay were incubated in situ, beginning on d 10, for 0, 2, 4, 8, 12, 16, 24, 36, 48, 72, and 96 h . Ruminal fluid was collected and pH recorded at -2, 0, 2, 4, 6, 8, 10, and 12 h after feeding on d 13 . Duodenal samples were taken for three consecutive days beginning on d 10 in a manner that allowed for a collection to take place every other hour over a 24-h period . Linear, quadratic, and cubic contrasts were used to evaluate the effects of increasing field pea level . Total DMI and OMI increased quadratically (P = 0.09), whereas forage DMI decreased quadratically (P = 0.09) with increasing field pea supplementation . There was a cubic effect (P < 0.001) for ruminal pH . Ruminal (P = 0.02) and apparent total-tract (P = 0.09) NDF disappearance decreased linearly with increasing field pea supplementation . Total ruminal VFA concentrations responded cubically (P = 0.008) . Bacterial N flow (P = 0.002) and true ruminal N disappearance (P = 0.003) increased linearly, and apparent total-tract N disappearance increased quadratically (P = 0.09) with increasing field pea supplementation . No treatment effects were observed for ruminal DM fill (P = 0.82), true ruminal OM disappearance (P = 0.38), apparent intestinal OM digestion (P = 0.50), ruminal ADF disappearance (P = 0.17), apparent total-tract ADF disappearance (P = 0.35), or in situ DM disappearance of forage (P = 0.33) . Because of effects on forage intake and ruminal pH, field peas seem to act like cereal grain supplements when used as supplements for forage-based diets . Supplementing field peas seems to effectively increase OM and N intakes of moderate-quality grass hay diets.

J Anim Sci, 2004 Jul, 82(7), 2123 - 30
Effect of field pea level on intake, digestion, microbial efficiency, ruminal fermentation, and in situ disappearance in beef steers fed growing diets; Reed JJ et al.; Effects of increasing level of field pea (variety: Profi) on intake, digestion, microbial efficiency, and ruminal fermentation were evaluated in beef steers fed growing diets . Four ruminally and duodenally cannulated crossbred beef steers (367+/-48 kg initial BW) were used in a 4 x 4 Latin square . The control diet consisted of 50% corn, 23% corn silage, 23% alfalfa hay, and 4% supplement (DM basis) . Treatments were field pea replacing corn at 0, 33, 67, or 100% . Diets were formulated to contain a minimum of 12% CP, 0.62% Ca, 0.3% P, and 0.8% K (DM basis) . Each period was 14 d long . Steers were adapted to the diets for 9 d . On d 10 to 14, intakes were measured . Field pea was incubated in situ, beginning on d 10, for 0, 2, 4, 8, 12, 16, 24, 36, 48, 72, and 96 h . Bags were inserted in reverse order, and all bags were removed at 0 h . Ruminal fluid was collected and pH recorded at -2, 0, 2, 4, 6, 8, 10, and 12 h after feeding on d 13 . Duodenal samples were taken for three consecutive days beginning on d 10 in a manner that allowed for a collection to take place every other hour over a 24-h period . Linear, quadratic, and cubic contrasts were used to compare treatments . There were no differences in DMI (12.46 kg/d, 3.16% BW; P > 0.46) . Ruminal dry matter fill (P = 0.02) and mean ruminal pH (P = 0.009) decreased linearly with increasing field pea level . Ruminal ammonia-N (P < 0.001) and total VFA concentrations (P = 0.01) increased linearly with increasing field pea level . Total-tract disappearance of OM (P = 0.03), N (P = 0.01), NDF (P = 0.02), and ADF (P = 0.05) increased linearly with an increasing field pea level . There were no differences in total-tract disappearance of starch (P = 0.35) . True ruminal N disappearance increased linearly (P < 0.001) with increasing field pea level . There were no differences in ruminal disappearance of OM (P = 0.79), starch (P = 0.77), NDF (P = 0.21), or ADF (P = 0.77) . Treatment did not affect microbial efficiency (P = 0.27) . Field pea is a highly digestible, nutrient-dense legume grain that ferments rapidly in the rumen . Because of their relatively high level of protein, including field peas in growing diets will decrease the need for protein supplementation . Based on these data, it seems that field pea is a suitable substitute for corn in growing diets.

Extremophiles, 2004 Aug, 8(4), 269 - 82 Epub 2004 Apr 09.
Geochemical and microbiological evidence for a hydrogen-based, hyperthermophilic subsurface lithoautotrophic microbial ecosystem (HyperSLiME) beneath an active deep-sea hydrothermal field; Takai K et al.; Subsurface microbial communities supported by geologically and abiologically derived hydrogen and carbon dioxide from the Earth's interior are of great interest, not only with regard to the nature of primitive life on Earth, but as potential analogs for extraterrestrial life . Here, for the first time, we present geochemical and microbiological evidence pointing to the existence of hyperthermophilic subsurface lithoautotrophic microbial ecosystem (HyperSLiME) dominated by hyperthermophilic methanogens beneath an active deep-sea hydrothermal field in the Central Indian Ridge . Geochemical and isotopic analyses of gaseous components in the hydrothermal fluids revealed heterogeneity of both concentration and carbon isotopic compositions of methane between the main hydrothermal vent (0.08 mM and -13.8 per thousand PDB, respectively) and the adjacent divergent vent site (0.2 mM and -18.5 per thousand PDB, respectively), representing potential subsurface microbial methanogenesis, at least in the divergent vent emitting more 13C-depleted methane . Extremely high abundance of magmatic energy sources such as hydrogen (2.5 mM) in the fluids also encourages a hydrogen-based, lithoautotrophic microbial activity . Both cultivation and cultivation-independent molecular analyses suggested the predominance of Methanococcales members in the superheated hydrothermal emissions and chimney interiors along with the other major microbial components of Thermococcales members . These results imply that a HyperSLiME, consisting of methanogens and fermenters, occurs in this tectonically active subsurface zone, strongly supporting the existence of hydrogen-driven subsurface microbial communities.

Am J Physiol Gastrointest Liver Physiol, 2004 Dec, 287(6), G1168 - 74 Epub 2004 Aug 12.
Butyrate specifically modulates MUC gene expression in intestinal epithelial goblet cells deprived of glucose; Gaudier E et al.; The mucus layer covering the gastrointestinal mucosa is considered the first line of defense against aggressions arising from the luminal content . It is mainly composed of high molecular weight glycoproteins called mucins . Butyrate, a short-chain fatty acid produced during carbohydrate fermentation, has been shown to increase mucin secretion . The aim of this study was to test 1) whether butyrate regulates the expression of various MUC genes, which are coding for protein backbones of mucins, and 2) whether this effect depends on butyrate status as the major energy source of colonocytes . Butyrate was provided at the apical side of human polarized colonic goblet cell line HT29-Cl.16E in glucose-rich or glucose-deprived medium . In glucose-rich medium, butyrate significantly increased MUC3 and MUC5B expression (1.6-fold basal level for both genes), tended to decrease MUC5AC expression, and had no effect on MUC2 expression . In glucose-deprived medium, i.e., when butyrate was the only energy source available, MUC3 and MUC5B increase persisted, whereas MUC5AC expression was significantly enhanced (3.7-fold basal level) and MUC2 expression was strikingly increased (23-fold basal level) . Together, our findings show that butyrate is able to upregulate colonic mucins at the transcriptional level and even better when it is the major energy source of the cells . Thus the metabolism of butyrate in colonocytes is closely linked to some of its gene-regulating effects . The distinct effects of butyrate according to the different MUC genes could influence the composition and properties of the mucus gel and thus its protective function.

Appl Biochem Biotechnol, 1999 Aug, 81(2), 119 - 30
Xylitol Production from Wood Hydrolyzates by Entrapped Debaryomyces hansenii and Candida guilliermondii Cells; Dominguez JM et al.; Debaryomyces hansenii cells were entrapped in Ca-alginate beads and used for producing xylitol from wood hydrolyzates . Batch experiments showed that bioconversion was severely hindered when Ca-alginate beads were hardened with Al3+ solutions . As an alternative to Al3+ hardening, the improvements in both mechanical stability of bioparticles and fermenting ability of the immobilized system derived from using increased concentrations of sodium alginate were assessed . The best results were obtained using a 4% (w/v) Na-alginate solution in the gelification step . This concentration was selected to perform continuous fermentations in a packed-bed reactor using raw or charcoal-treated hydrolyzates (15.5 g of xylose/L) with two different yeasts: Candida guilliermondii and Debaryomyces hansenii . With a final cell concentration of about 50 g of cells/L (0.075 g of cells/g of beads), the volumetric productivities reached with these yeasts in media made from charcoal-treated hydrolyzates were 0.58 and 0.91 g/L.h, respectively.

Appl Biochem Biotechnol, 2004 Jun, 117(3), 155 - 64
Screening of laccase, manganese peroxidase, and versatile peroxidase activities of the genus pleurotus in media with some raw plant materials as carbon sources; Stajic M et al.; Species of the genus Pleurotus are among the most efficient natural species in lignin degradation belonging to the subclass of ligninolytic organisms that produce laccase (Lac), Mn-dependent peroxidase (MnP), versatile peroxidase (VP), and the H2O2-generating enzyme aryl-alcohol oxidase, but not lignin peroxidases . Production of Lac and oxidation of 2,6-dimethoxyphenol (DMP) in the presence and absence of Mn2+ were detected both in submerged fermentation (SF) of dry ground mandarine peels and in solid-state fermentation (SSF) of grapevine sawdust in all investigated Pleurotus species and strains . Evidence of cultivation methods having a distinct influence on the level of enzyme activities has been demonstrated . Most of the species and strains had higher Lac activity under SSF conditions than under SF conditions . DMP oxidation in the presence and absence of Mn2+ was detected in all investigated species and strains, but was lower under SF conditions than under SSF conditions for most of them . However, relative activities of DMP oxidation in the absence of Mn2+, as percentages of activity against DMP in the presence of Mn2+, were higher under conditions of SF than in SSF cultures in most of the investigated species and strains . The obtained results showed that strains of different origins have different efficiently ligninolytic systems and that conditions of SSF are more favorable for ligninolytic activity than those in SF owing to their similarity to natural conditions on wood substrates . Copryright 2004 Humana Press Inc.

Appl Biochem Biotechnol, 2004 Jul-Sep, 118(1-3), 337 - 48
Effect of vitamin E on autolysis and sporulation of Aspergillus nidulans; Emri T et al.; The morphologic and physiologic effects of vitamin E, a powerful antioxidant, on the autolysis and sporulation of Aspergillus nidulans FGSC26 were studied . In carbon-depleted submerged cultures, reactive oxygen species (ROS) accumulated in the cells and, concomitantly, progressing autolysis was observed, which was characterized by decreasing dry cell masses and pellet diameters as well as by increasing extracellular chitinase activities . Vitamin E supplemented at a concentration of 1 g/L hindered effectively the intracellular accumulation of ROS, the autolytic loss of biomass, the disintegration of pellets, and the release of chitinase activities . In surface cultures, vitamin E inhibited autolysis of both A . nidulans FGSC26 and a loss-of-function FlbA autolytic phenotype mutant . In addition, supplementation of the culture medium with this antioxidant also had a negative effect on the sporulation of strain FGSC26 and the FadAG203R hypersporulating phenotype mutant . These results suggest that accumulation of ROS was involved in the initiation of both sporulation and autolysis in this filamentous fungus, but that FadA/FlbA signaling was not involved in this vitamin E-dependent regulation . Vitamin E can be recommended as a supplement in fermentations in which the disintegration of pellets and gross autolysis should be avoided.

Appl Biochem Biotechnol, 2004 Jul-Sep, 118(1-3), 189 - 204
Production of chitinolytic enzymes with Trichoderma longibrachiatum IMI 92027 in solid substrate fermentation; Kovacs K et al.; Thirty Trichoderma strains representing 15 species within the genus were screened for extracellular production of chitinolytic enzymes in solid substrate fermentation . Trichoderma longibrachiatum IMI 92027 (ATCC 36838) gave the highest yield (5.0 IU/g of dry matter of substrate) after 3 d of fermentation on wheat bran-crude chitin (9:1 mixture) medium . The optimal moisture content (66.7%), chitin content (20%), initial pH of the medium (2.0-5.0), and time course (5 d) of solid substrate fermentation were determined for strain IMI 92027 . Cellulase, xylanase, alpha-amylase, and beta-xylosidase activities were also detected . The pH and temperature optima of the chitinase complex of T . longibrachiatum IMI 92027 were 4.5 and 55 degrees C, respectively . The enzyme totally lost its activity at 70 degrees C in 5 min in the absence of the substrate but retained about 15% of its initial activity even at 70 degrees C after a 60-min incubation in the presence of solid substrate fermentation solids . Purification of protein extract from the solid substrate fermentation material revealed high chitinolytic activities between pI 5.9 and 4.8, where N-acetyl-beta-D-hexosaminidase and chitinase peaks have been found in the same pI range . Two chitinases of 43.5 and 30 kDa were purified at acidic pI.

Appl Biochem Biotechnol, 2004 Jul-Sep, 118(1-3), 81 - 8
Development of a bionematicide with Paecilomyces lilacinus to control Meloidogyne incognita; Brand D et al.; Root-knot disease caused by Meloidogyne incognita is a matter of grave concern because it affects several economically important crop plants . The use of solid-state fermentation (SSF) may help to elaborate efficient formulations with fungi to be employed in the biologic control of nematodes . Attempts were made to select low-cost substrates for spore production of a strain of Paecilomyces lilacinus with known nematicide capacity . Coffee husks, cassava bagasse, and defatted soybean cake were utilized as substrates, and sugarcane bagasse was used as support . Fermentations were carried out in flasks covered with filter paper at 28 degrees C for 10 d . The products obtained by SSF were evaluated for their nematicide activity in pot experiments containing one seedling of the plant Coleus inoculated with the nematode M . incognita . The plants were evaluated 2 mo after inoculation . Fermented products showed a reduction in the number of nematodes . The best results were obtained with defatted soybean cake, which showed almost 100% reduction in the number of nematodes; the reduction with coffee husk was 80% and with cassava bagasse was about 60%.

Appl Biochem Biotechnol, 2004 Jul-Sep, 118(1-3), 73 - 9
Enzyme production and profile by Aspergillus niger during solid substrate fermentation using palm kernel cake as substrate; Ong LG et al.; The oil palm sector is one of the major plantation industries in Malaysia . Palm kernel cake is a byproduct of extracted palm kernel oil . Mostly palm kernel cake is wasted or is mixed with other nutrients and used as animal feed, especially for ruminant animals . Recently, palm kernel cake has been identified as an important ingredient for the formulation of animal feed, and it is also exported especially to Europe, South Korea, and Japan . It can barely be consumed by nonruminant (monogastric) animals owing to the high percentages of hemicellulose and cellulose contents . Palm kernel cake must undergo suitable pretreatment in order to decrease the percentage of hemicellulose and cellulose . One of the methods employed in this study is fermentation with microorganisms, particularly fungi, to partially degrade the hemicellulose and cellulose content . This work focused on the production of enzymes by Aspergillus niger and profiling using palm kernel cake as carbon source.

Appl Biochem Biotechnol, 2004 Jul-Sep, 118(1-3), 33 - 46
Evolutionary operation-factorial design technique for optimization of conversion of mixed agroproducts into gallic acid; Mukherjee G et al.; This article presents the optimization of gallic acid production using filamentous fungi from tannin-rich mixed substrates taking into account the interaction effects of six variable process parameters . The methodology adopted for optimization was the evolutionary operation (EVOP)-factorial design technique . This technique combines the factorial method for designing experiments with the EVOP methodology for analyzing the experimental results systematically and arriving at conclusions according to its decision-making procedure . Standard deviation and error limits based on 95% confidence were calculated according to the relationship given in the literature . It was found that the best combinations of the process parameters at the optimum levels were 30 degrees C, 80% relative humidity, pH 5.0, 48-h incubation period, 3 mL of induced inoculum, and 35 g of mixed substrate, resulting in a gallic acid yield of 94.8% under modified solid-state fermentation.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 571 - 6
A preliminary information about continuous fermentation using cell recycling for improving microbial xylitol production rates(scientific note); Silva SS et al.; Xylitol is a sugar-alcohol with important technological properties, such as anticariogenicity, low caloric value, and negative dissolution heat . It can be used successfully in food formulations and pharmaceutical industries . Its production is therefore in great demand . Biotechnological xylitol production has several economic advantages in comparison with the conventional process based on the chemical reduction of xylose . The efficiency and the productivity of this fermentation chiefly depends on the microorganism and the process conditions employed . In this article a simple continuous culture with cell recycling was evaluated to enhance the capability of Candida guilliermondii FTI 20037 to produce xylitol . The fermentation was initiated batchwise by directly inoculating the grown seed culture in a 2-L bench-scale fermentor . Continuous feeding was begun at a dilution rate (D) of 0.060/h after the xylose concentration had completely consumed and the cell concentration was about 4.0 g/L . At a dilution rate of 0.060/h the xylitol concentration was about 15 g/L and increased by about 35%, whereas the dilution rate decreased by about 58% . Furthermore, the volumetric productivity, Qp, markedly depended on the dilution rate, diminishing by about 37% as D was changed from 0.060 to 0.025/h . These preliminary results show us that continuous fermentation with cell recycling is a good way to study the xylitol production by xylose-fermenting yeasts.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 547 - 60
Ethanol production using concentrated oak wood hydrolysates and methods to detoxify; Lee WG et al.; Ethanol production from concentrated oak wood hydrolysate was carried out to obtain a high ethanol concentration and a high ethanol yield . The effect of added inhibitory compounds, which are typically produced in the pretreatment step of steam-explosion on ethanol fermentation, was also examined . p-Hydroxybenzoic aldehyde, a lignin-degradation product, was the most inhibitory compound tested in this study . Compounds with additional methyl groups had reduced toxicity and the aromatic acids were less toxic than the corresponding aldehydes . The lignin-degradation products were more inhibitory than the sugar-derived products, such as furfural and 5-hydroxymethylfurfural (HMF) . Adaptation of yeast cells to the wood hydrolysate and detoxification methods, such as using charcoal and overlime, had some beneficial effects on ethanol production using the concentrated wood hydrolysate . After treatment with charcoal and low-temperature sterilization, the yeast cells could utilize the concentrated wood hydrolysate with 170 as well as 140 g/L glucose, and produce 69.9 and 74.2 g/L ethanol, respectively, with a yield of 0.46-0.48 g ethanol/g glucose . In contrast, the cells could not completely utilize untreated wood hydrolysate with 100 g/L glucose . Low-temperature sterilization, with or without charcoal treatment, was very effective for ethanol production when highly concentrated wood hydrolysates were used . Low-temperature sterilization has advantages over traditional detoxification methods, such as using overlime, ion exchange, and charcoal, because of the reduction in the total cost of ethanol production.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 535 - 46
Ethanol Production by a Flocculant Yeast Strain in a CSTR Type Fermentor with Cell Recycling; Hojo O et al.; Tests were performed in a continuous stirred tank reactor (CSTR), with and without cell recycling, to produce ethanol . The reactor without cell recycling produced the kinetic model of ethanol production, whereas the reactor with cell recycling allowed for a study of process stability . The Levenspiel kinetic model was adopted; however, in the case of fermentation with cell recycling, the coefficient of cell death was added . It was observed that cellular viability varied greatly throughout the fermenting process and that microaeration is of fundamental importance in maintaining the stability of the process.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 485 - 500
Electrodialysis of acetate fermentation broths; Chukwu UN et al.; Electrodialysis (ED) shows good potential for downstream processing of acetate fermentation broths, to separate acetic acid while unreacted glucose and other nutrients are partially recycled back to the fermenter . With conventional anion- and cation-exchange membranes, higher current increased acetate flux, water flux, and energy consumption . Multiple ED stacks connected in series with unequal initial volumes for a batch process maximized acetate concentration in the concentrating stream to 134 g/L calcium-magnesium acetate (CMA) in the fermentation broth at pH 6.8 . Back-transport of acetate from the product into the feed stream and water transport limit the maximum concentration possible . Cost of ED is about $295/ton acetate for the CMA broth.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 473 - 84
Separation optimization for the recovery of phenyl ethyl alcohol; Priddy SA et al.; Phenyl ethyl alcohol is a compound that occurs naturally in flower petals and in many common beverages, such as beer . Desire for the floral, rose-like notes imparted by phenyl ethyl alcohol has created a unique niche for this chemical in flavor and fragrance industries . Phenyl ethyl alcohol can be produced by Saccharomyces cerevisiae via bioconversion . Often this method of production results in extremely low yields, thus placing a great deal of importance on recovery and purification of the valuable metabolite . To determine the best method for recovering the chemical, a primary recovery step and a secondary recovery step were developed . The primary recovery step consisted of comparing dead-end filtration with crossflow ultrafiltration . Crossflow ultrafiltration was ultimately selected to filter the fermentation broth because of its high flow rates and low affinity for the product . The secondary recovery step consisted of a comparison of liquid- liquid extraction and hydrophobic resin recovery . The hydrophobic resin was selected because of its higher rate of recovery and a higher purity than the liquid-liquid extraction, the current practice of Brown-Forman.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 435 - 44
Bioconversion of secondary fiber fines to ethanol using counter-current enzymatic saccharification and co-fermentation; Jeffries TW et al.; This research examined several enzymatic and microbial process for the conversion of waste cellulosic fibers into ethanol . The first was a one-stage process in which pulp fines were contacted with commercial enzyme solutions . The second process used sequential, multistage saccharification . The third used sequential enzyme addition in a countercurrent mode . Experiments compared the results with various feedstocks, different commercial enzymes, supplementation with beta-glucosidase, and saccharification combined with fermentation . The highest saccharification (65%) from a 4% consistency pulp and the highest sugar concentration (5.4%) from an 8% consistency pulp were attained when 5 FPU/g plus 10 IU/g of beta-glucosidase were used . Sequential addition of enzyme to the pulp in small aliquots produced a higher overall sugar yield/U enzyme than the addition of the same total amount of enzyme in a single dose . In the saccharification and fermentation experiments, we produced 2.12% ethanol from a 5.4% sugar solution . This represents 78% of the theoretical maximum . This yield could probably be increased through optimization of the fermentation step . Even when little saccharification occurred, the enzyme facilitated separation of water, fiber, and ash, so cellulase treatment could be an effective means for dewatering pulp sludges.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 401 - 8
Optimization of L-Lactic Acid Production from Glucose by Rhizopus oryzae ATCC 52311; Zhou Y et al.; The effect of nutrients on L(+)-lactic acid production from glucose was investigated using Rhizopus oryzae ATCC 52311 . From the shake-flask experiments, the optimal medium composition was defined for improved lacticacid production . In order to enhance lactic-acid production rate and product yield, controlled aeration in a bubble column was conducted under optimal conditions . Results showed a maximum lactic-acid production rate of 2.58 g/L/h was obtained with an initial glucose concentration of 94 g/L . Final lactic-acid concentration of 83 g/L was achieved after 32 h of fermentation with a weight of 0.88 g lactic acid/g glucose consumed.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 373 - 88
Fermentation Kinetics of Ethanol Production from Glucose and Xylose by Recombinant Saccharomyces 1400(pLNH33); Krishnan MS et al.; Fermentation kinetics of ethanol production from glucose, xylose, and their mixtures using a recombinant Saccharomyces 1400(pLNH33) are reported . Single-substrate kinetics indicate that the specific growth rate of the yeast and the specific ethanol productivity on glucose as the substrate was greater than on xylose as a substrate . Ethanol yields from glucose and xylose fermentation were typically 95 and 80% of the theoretical yield, respectively . The effect of ethanol inhibition is more pronounced for xylose fermentation than for glucose fermentation . Studies on glucose-xylose mixtures indicate that the recombinant yeast co-ferments glucose and xylose . Fermentation of a 52.8 g/L glucose and 56.3 g/L xylose mixture gave an ethanol concentration of 47.9 g/L after 36 h . Based on a theoretical yield of 0.51 g ethanol/g sugars, the ethanol yield from this experiment (for data up to 24 h) was calculated to be 0.46 g ethanol/g sugar or 90% of the theoretical yield . The specific growth rate of the yeast on glucose-xylose mixtures was found to lie between the specific growth rate on glucose and the specific growth rate on xylose . Kinetic studies were used to develop a fermentation model incorporating the effects of substrate inhibition, product inhibition, and inoculum size . Good agreements were obtained between model predictions and experimental data from batch fermentation of glucose, xylose, and their mixtures.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 867 - 76
The nature of lignin from steam explosion/ enzymatic hydrolysis of softwood: structural features and possible uses(scientific note); Shevchenko SM et al.; Effective utilization of the lignin by-product is a prerequisite to the commercial viability of ethanol production from softwood wastes using a steam explosion (SE)/enzymatic hydrolysis (EH)/fermentation process . Changes in the chemical composition of Douglas fir wood on SO2-catalyzed SE followed by EH were assessed using conventional analytical methods and new halogen-probe techniques . A significant solubilization of hemicelluloses was observed in the SE stage, the severity of which affected subsequent fermentation of cellulose and sorption of enzymes . SE of softwood resulted in dramatic changes in the chemical structure of lignin in the residual material involving chemical reactions via the benzyl cation . This leads to a more condensed lignin with partly blocked alpha-reaction centres . Possible uses for this lignin are discussed.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 845 - 56
Mathematical Modeling of Lipase and Protease Production by Penicillium restrictum in a Batch Fermenter; Freire DM et al.; This work presents a mathematical model that describes time course variations of extracellular lipase and protease activities for the batch fermentation of the fungus Penicillium restrictum, a new and promising strain isolated from soil and wastes of a Brazilian babassu coconut oil industry . The fermentation process was modeled by an unstructured model, which considered the following dependent variables: cells, fat acid, dissolved oxygen concentrations, lipase and protease activities, and cell lysate concentration . The last variable represents the amount of cells that has been lysed by the shear stress and natural cell death . Proteases released to the medium, as consequence of this process, enhance lipase inactivation . The model is able to predict the effects of some operation variables such as air flow rate and agitation speed . The mathematical model was validated against batch-fermentation data obtained under several operating conditions . Because substrate concentration has antagonistic effects on lipase activity, a typical optimization scheme should be developed in order to minimize these deleterious effects while maximizing lipase activity.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 735 - 44
Lignin Peroxidase and Protease Production by Streptomyces viridosporus T7A in the Presence of Calcium Carbonate Nutritional and Regulatory Carbon Sources; Macedo JM et al.; Streptomyces are good producers of enzymes of industrial interest, such as lignin peroxidase (LiP) and proteases . To optimize production of these enzymes by Streptomyces viridosporus T7A, two parameters were evaluated: carbon sources and calcium carbonate . Shake-flask fermentations were performed using culture media, with and without CaCO3, contained yeast extract, mineral salts and either glucose, lactose, galactose, or corn oil . In the absence of calcium carbonate, the maximum values for LiP and protease activities occurred during the idiophase with LiP activity being favored by glucose, corn oil, and galactose, and protease activity being favored only by corn oil . Calcium carbonate affected the cell morphology by reducing the size of the pellets . Moreover, in the presence of the salt, LiP production was growth-associated in all media but the glucose medium . Higher enzyme levels were observed when galactose and glucose were used as carbon sources . Protease activity was repressed by both glucose and galactose, whereas corn oil was the best carbon source for the enzyme production . Calcium carbonate increased LiP production by up to 2.6-fold . Such improvement was not observed for protease production, suggesting a selective effect of CaCO3 on LiP activity.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 633 - 40
Economic Analysis of Selected Lignocellulose-to-Ethanol Conversion Technologies; So KS et al.; The objective of this case study was to examine the economics of three lignocellulose-to-ethanol conversion technologies: fast pyrolysis integrated with a fermentation step, simultaneous saccharification and fermentation (SSF), and dilute sulfuric acid hydrolysis and fermentation . All technologies were assumed to have an annual production rate of 25 million gallons of ethanol . The three technologies were compared in terms of capital costs, operating costs, and ethanol production costs . Sensitivity analyses were carried out to study the uncertainties of wood costs and ethanol production rates on ethanol production costs . Final economic analysis showed that fast pyrolysis integrated with a fermentation step is comparable with the other two processes and suggests that it should be considered for further development.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 609 - 32
Biomass Conversion to Mixed Alcohol Fuels Using the MixAlco Process; Holtzapple MT et al.; The MixAlco process is a patented technology that converts any biodegradable material (e.g., sorted municipal solid waste, sewage sludge, industrial biosludge, manure, agricultural residues, energy crops) into mixed alcohol fuels containing predominantly 2-propanol, but also higher alcohols up to 7-tridecanol . The feedstock is treated with lime to increase its digestibility . Then, it is fed to a fermentor in which a mixed culture of acid-forming microorganisms produces carboxylic acids . Calcium carbonate is added to the fermentor to neutralize the acids to their corresponding carboxylate salt . The dilute (~3%) carboxylate salts are concentrated to 19% using an amine solvent that selectively extracts water . Drying is completed using multi-effect evaporators . Finally, the dry salts are thermally converted to ketones which subsequently are hydrogenated to alcohols . All the steps in the MixAlco process have been proven at the laboratory scale . A techno-economic model of the process indicates that with the tipping fees available in New York ($126/dry tonne), mixed alcohol fuels may be sold for $0.04/L ($0.16/gal) with a 60% return on investment (ROI) . With the average tipping fee in the United States rates ($63/dry tonne), mixed alcohol fuels may be sold for $0.18/L ($0.69/gal) with a 15% ROI . In the case of sugarcane bagasse, which may be obtained for about $26/dry ton, mixed alcohol fuels may be sold for $0.29/L ($1.09/gal) with a 15% ROI.

Appl Biochem Biotechnol, 1999 Spring, 77-79, 595 - 608
A process economic approach to develop a dilute-Acid cellulose hydrolysis process to produce ethanol from biomass; Nagle N et al.; Successful deployment of a bioethanol process depends on the integration of technologies that can be economically commercialized . Pretreatment and fermentation operations of the traditional enzymatic bioethanol-production process constitute the largest portion of the capital and operating costs . Cost reduction in these areas, through improved reactions and reduced capital, will improve the economic feasibility of a large-scale plant . A technoeconomic model was developed using the ASPEN Plus trade mark modeling software package . This model included a two-stage pretreatment operation with a co-current first stage and countercurrent second stage, a lignin adsorption unit, and a cofermentation unit . Data from kinetic modeling of the pretreatment reactions, verified by bench-scale experiments, were used to create the ASPEN Plus base model . Results from the initial pretreatment and fermentation yields of the two-stage system correlated well to the performance targets established by the model . The ASPEN Plus model determined mass and energy-balance information, which was supplied to an economic module to determine the required selling price of the ethanol . Several pretreatment process variables such as glucose yield, liquid: solid ratio, additional pretreatment stages, and lignin adsorption were varied to determine which parameters had the greatest effect on the process economics . Optimized values for these key variables became target values for the bench-scale research, either to achieve or identify as potential obstacles in the future commercialization process . Results from this modeling and experimentation sequence have led to the design of an advanced two-stage engineering- scale reactor for a dilute-acid hydrolysis process.

Appl Biochem Biotechnol, 1999 Apr, 80(1), 77 - 90
Effect of the Shear Rate on Pullulan Production from Beet Molasses by Aureobasidium pullulans in an Airlift Reactor; Roukas T et al.; The effect of the shear rate on pullulan production from beet molasses by Aureobasidium pullulans P56 in an airlift reactor was investigated . A maximum polysaccharide concentration (18.5 g/L), biomass dry weight (14.0 g/L), polysaccharide yield (38.5%), and sugar utilization (96%) was achieved at a shear rate of 42 s-1 . A . pullulans grown on beet molasses produced a mixture of pullulan and other polysaccharides . The highest value of pullulan proportion (30% of total polysaccharide) was obtained at a low shear rate (42 s-1) . The apparent viscosity of the fermentation broth increased as the shear rate increased up to 42 s-1 and then decreased . On the other hand, the dissolved oxygen concentration and the volumetric mass transfer coefficient increased with the increase of the shear rate from 21 to 84 s-1 . The external addition of L-glutamic acid, olive oil, and Tween-80 improved significantly the production of crude polysaccharide (27.0 g/L), but the pullulan content of the polysaccharide was low (20%).

Appl Biochem Biotechnol, 1999 Apr, 80(1), 65 - 76
Effect of Fermentation Conditions in the Enzymatic Activity and Stereoselectivity of Crude Lipase from Candida rugosa; Sanchez A et al.; Different fed-batch cultures of Candida rugosa were carried out using oleic acid as the only carbon source . The crude lipases obtained under several operational conditions and downstream processes showed different catalytic activity and isoenzymes ratio . This fact implied that the performance of the lipase produced could be modulated by using different operational fermentation conditions . These powders were compared with commercial lipase from Sigma (St . Louis, MO) in hydrolysis and synthesis reactions . Especially interesting was the fact that the enantioselectivity of a crude lipase was higher than that observed with commercial lipase in the resolution of racemic Ketoprofen . In addition, response of both lipases in the presence of water was different.

Bioprocess Biosyst Eng, 2004 Oct, 26(5), 331 - 40 Epub 2004 Aug 06.
Computer control of pH and DO in a laboratory fermenter using a neural network technique; Meszaros A et al.; In this contribution, the advantages of the artificial neural network approach to the identification and control of a laboratory-scale biochemical reactor are demonstrated . It is very important to be able to maintain the levels of two process variables, pH and dissolved oxygen (DO) concentration, over the course of fermentation in biosystems control . A PC-supported, fully automated, multi-task control system has been designed and built by the authors . Forward and inverse neural process models are used to identify and control both the pH and the DO concentration in a fermenter containing a Saccharomyces cerevisiae based-culture . The models are trained off-line, using a modified back-propagation algorithm based on conjugate gradients . The inverse neural controller is augmented by a new adaptive term that results in a system with robust performance . Experimental results have confirmed that the regulatory and tracking performances of the control system proposed are good.

Appl Microbiol Biotechnol . 2004 Aug 5; {Epub ahead of print}
Improvement of arachidonic acid production by mutants with lower n-3 desaturation activity derived from Mortierella alpina 1S-4; Sakuradani E et al.; Five mutants were obtained, Y11, Y135, Y164, Y180 and Y61, capable of accumulating higher amounts of arachidonic acid (AA) than Mortierella alpina 1S-4, an industrial strain for the production of AA-rich triacylglycerol (TG) . This is thought to be due to low or no activity of n-3 desaturation with conversion of AA to eicosapentaenoic acid, which functions at a cultural temperature below 20 degrees C . In small-scale cultivation under optimum conditions, Y11 and Y61 respectively accumulated 4.97 mg/ml and 4.11 mg/ml of AA, using a high concentration of glucose at 20 degrees C, compared with 3.74 mg/ml for M . alpina 1S-4 . In a 5-l jar fermentor, the AA content in Y11 and Y61 kept increasing during cultivation, with consumption of the glucose in the medium; and this reached 1.48 mg/ml and 1.77 mg/ml (118 mg/g, 120 mg/g of dry mycelia) at day 10, respectively, compared with 0.95 mg/ml (86 mg/g of dry mycelia) for M . alpina 1S-4 . From the results of lipid analysis, the TG contents of Y11 and Y61 in the major lipids were significantly higher than that of M . alpina 1S-4; and the AA percentages in TG of Y11 and Y61 were also higher . Both Y11 and Y61 are potential producers of TG rich in AA.

Medicina (Kaunas), 2004, 40(8), 758 - 61
{The analysis of tincture for improvement of blood circulation}; Bernatoniene R et al.; The article describes qualitative and quantitative analysis of tincture for improvement of blood circulation . Flavonoids have been qualitatively determined by methods of thin-layer chromatography, high-pressure liquid chromatography and color reaction with magnesium powder and concentrated hydrochloric acid . Ferments were identified with ferric ammonium sulphate solution; glycosides--with dimethylaminobenzaldehyde solution in sulphur acid; saponin--with lead subacetate solution; reductive materials - with silver nitrate ammoniacal solution; albumen--with ninhydrin solution; and caffeic acid--by method of thin-layer chromatography . An optimal spectrophotometrical method for determining the quantity of quercetin was applied . The refractive index, relative density, loss of drying and ethanol concentration were determined according to European Pharmacopoeia Requirements.

Curr Microbiol, 2004 Aug, 49(2), 75 - 8
Anaerobic 2-propanol degradation in anoxic paddy soil and the possible role of methanogens in its degradation; Tonouchi A; The anaerobic degradation of 2-propanol in anoxic paddy soil was studied with soil cultures and a 2-propanol-utilizing methanogen . Acetone was the first and the major intermediate involved in the methanogenic degradation of 2-propanol . Analyses with a methanogenesis inhibitor, bacteria antibiotics, and the addition of H2 to the gas phase revealed that 2-propanol oxidation to acetone directly occurred using 2-propanol-utilizing methanogens, but not with H2-producing syntrophic bacteria, for which the removal of acetone is required for complete 2-propanol oxidation . The 2-propanol-utilizing strain IIE1, which is phylogenetically closely related to Methanoculleus palmolei, was isolated from paddy soil, and the potential role of the strain in 2-propanol degradation was investigated . 2-Propanol is one of the representative fermentation intermediates in anaerobic environments . This is the first report on the anaerobic 2-propanol degradation process.

Exp Anim, 2004 Jul, 53(4), 361 - 71
Effect of sugar alcohols on gut function and body composition in normal and cecectomized rats; Islam MS et al.; The effects of two sugar alcohols on feed utilization, digesta retention, gut fermentation and serum lipid profiles were compared in normal and cecectomized rats to examine the possibility of the cecectomized rat as an experimental animal with relevance to humans . Semi-purified diets containing no sugar alcohol, 7% sorbitol or 7% lactitol were fed to normal and cecectomized rats for 16 days . The digestibility of the crude fat and the compositions of the carcass dry matter and crude fat were significantly decreased by feeding sugar alcohols in both groups, but the effects were relatively higher in the cecectomized rats than in the normal rats . Diarrhea, faster transit times and shorter retention times of digesta were noted in the cecectomized rats fed sugar alcohols, while the inverse results were observed in the normal rats fed similar diets . The concentration of cecal organic acids was increased in the normal rats, whereas the concentration of colonic organic acids was decreased in the cecectomized rats fed sugar alcohols, compared with their corresponding control groups . The concentration of serum total cholesterol was decreased in both the normal and cecectomized rats fed diets containing sugar alcohols . The tendencies for diarrhea, faster digesta transit and reduced body fat induced by the fermentable materials in the cecectomized rat have good relevance to the parallel effects of fermentable materials in humans, suggesting the possibility of using the cecectomized rat as a model to study some of the physiological effects of sugar alcohols in humans.

Clin Cancer Res, 2004 Aug 1, 10(15), 5282 - 92
A concentrated aglycone isoflavone preparation (GCP) that demonstrates potent anti-prostate cancer activity in vitro and in vivo; Bemis DL et al.; PURPOSE: Isoflavones have anticancer activities, but naturally occurring isoflavones are predominantly glycosylated and poorly absorbed . Genistein combined polysaccharide (GCP; Amino Up Chemical Co., Sapporo, Japan), is a fermentation product of soy extract and basidiomycetes mycillae that is enriched in biologically active aglycone isoflavones . This study analyzes GCP in vitro and in vivo for potential utility as a prostate cancer chemopreventative agent . EXPERIMENTAL DESIGN: Androgen-sensitive LNCaP and androgen-independent PC-3 cells were grown with various concentrations of GCP . In vitro cell growth was analyzed by the WST-1 assay, and apoptosis was assessed by fluorescence-activated cell sorting and detection of poly(ADP-ribose) polymerase cleavage using Western blot techniques . Effects of GCP on expression of cell cycle-regulatory proteins p53 (LNCaP only), p21, and p27 and the protein kinase Akt were considered using Western blot techniques . An in vivo LNCaP xenograft model was used to study the effects of a 2% GCP-supplemented diet on tumor growth in comparison with a control diet . RESULTS: GCP significantly suppressed LNCaP and PC-3 cell growth over 72 h (89% and 78% in LNCaP and PC-3, respectively, at 10 microg/ml; P < 0.0001) . This reduction was associated with apoptosis in LNCaP cells, but not in PC-3 cells . GCP induced p27 and p53 (LNCaP only) protein expression within 6 h and suppressed phosphorylated Akt in both cell lines . The 2% GCP-supplemented diet significantly slowed LNCaP tumor growth, increasing apoptosis (P < 0.001), and decreasing proliferation (P < 0.001) over 4 weeks . CONCLUSIONS: GCP has potent growth-inhibitory effects against prostate cancer cell lines in vitro and in vivo . These data suggest GCP has potential as an effective chemopreventive agent against prostate cancer cell growth.

Curr Opin Biotechnol, 2004 Aug, 15(4), 330 - 4
Detergent proteases; Maurer KH; Over the past 20 years, the development of subtilisins as typical detergent proteases has employed all the tools of enzyme technology, resulting in a constant flow of new and improved enzymes . The number of molecules identified and characterized, however, is in clear opposition to the number of molecules that are entering the market . Will the next-generation detergent proteases be based on new backbones different from subtilisins, or will the use of all available technologies (rational design, directed evolution and exploitation of natural diversity) yield improved subtilisins, ending the current era dominated by high alkaline subtilisins? These questions will have to be answered not only by the performance of the molecules themselves, but also by their yield in fermentation and their compatibility with existing production technologies.

Biotechnol Prog, 2004 Jul-Aug, 20(4), 1280 - 4
Solid-state fermentation for production of griseofulvin on rice bran using Penicillium griseofulvum; Saykhedkar SS et al.; Griseofulvin is a secondary metabolite produced from fungal species that have morphology suitable for solid-state fermentation (SSF) . Reports on production of griseofulvin by SSF are scarce . The present work investigates SSF for griseofulvin production, optimization of its process parameters vis-a-vis the conventional submerged fermentation and its downstream processing from the same . Rice bran adjusted to an initial moisture content (IMC) of 50% (v/w) inoculated with 1 mL of a suspension of 10(6) spores/mL under agitation at 250 rpm containing the modified Czapek-Dox medium and additional 0.1% choline chloride as a precursor gave a yield of griseofulvin in 9 days that was comparable to submerged fermentation after 28 days . The yield of griseofulvin (microg/g dry biomass) was comparable in SSF and submerged fermentation . The biomass was estimated by estimation of chitin . Discussions on the effect of each parameter in SSF have also been included.

Biotechnol Prog, 2004 Jul-Aug, 20(4), 1091 - 5
Development of a novel whey beverage by fermentation with kefir granules . Effect of various treatments; Athanasiadis I et al.; The development of a novel whey-based beverage with acceptable organoleptic properties is reported, where various treatments were studied . Kefir yeast immobilized on delignified cellulosic materials (DCM) or gluten pellets proved to accelerate whey fermentation significantly, with the latter support being not so preferable . Kefir granules seemed to achieve similar fermentation times as DCM . The final pH of the product is suggested to be 4.1 since the profile of the volatile byproducts was higher than other pH values tested . The addition of fructose seemed to be beneficial on the volatile content of the product, although its acceptability as determined by a preference panel was similar to that of the control . Finally, black raisin extract appeared to promote fermentation without any positive effect on the preference of the evaluators.

Commun Agric Appl Biol Sci, 2003, 68(2 Pt A), 327 - 30
Enzymatic conversion of the clavan exopolysaccharide by Streptomyces sp . YSDL-20; Yemmas L et al.; A screening programme was set up to isolate microorganisms able to hydrolyse the complex biopolymer clavan produced by Clavibacter michiganensis subsp . michiganensis LMG 5604 . This valuable exopolysaccharide is very rich in L-fucose (37.5% w/w), a rare sugar, used in the medical field (Vanhooren, 1999) . A microorganism capable of depolymerizing the polymer may decrease the high viscosity during clavan batch fermentations and remove the limitations of the oxygen transfer and consequently increase the clavan yield . It could also release free L-fucose or L-fucose rich oligosaccharides . An actinomycete, designated YSDL-20, isolated from a soil sample, was able to depolymerize this biopolymer . Based on its morphology and molecular characteristics, this strain could only be identified as Streptomyces sp. . On clavan, this strain displays good growth (17.5 g DCW/l after 96 h of cultivation) characterized by filamentous growth during the earlier days of cultivation followed by sporulation after 4 days . The flow behaviour of the Clavibacter broth was characterized, the fermentation culture broth behaves as a pseudoplastic fluid . The viscosity of the culture broth as well as of the purified clavan EPS, decreases when lyophilised supernatant of Streptomyces sp . YSDL-20 was added, indicating clavanase action . The viscosity decreases by 26% when the Clavibacter culture broth was incubated during 18 h with the crude Streptomyces enzyme source, whereas a 82% viscosity drop was observed, when the purified clavan EPS (10 g/l) was incubated with the lyophilised Streptomyces supernatant for 5 h.

Commun Agric Appl Biol Sci, 2003, 68(2 Pt A), 325 - 6
Microbial biosynthesis and making of pigment melanin; Vinarov A et al.; The efficient strain and the technology of biosynthesis and extraction process from a biomass received in process cultivation of new yeast strain Saccharomyces neoformans var . nigricans are developed . The influence of a carbon substrates (glucose, spirit, milk whey) and conditions of fermentation process on an output and chromatically received melanin are investigated . It is is established, that for the given strain the fermentation time makes 36-40 hours.

Commun Agric Appl Biol Sci, 2003, 68(2 Pt A), 313 - 6
Physiological properties and enzymatic activities of Rhizopus oligosporus in solid state fermentations; Miszkiewicz H et al.; The physiology of fungus Rhizopus oligosporus in the solid state fermentation of Pisum sativum was investigated by means of digital analysis of microscopic image . The correlations between the hyphal fractions within physiological zones, the release of glucose and soluble proteins, and enzymatic activities of the examined strain were also estimated.

Commun Agric Appl Biol Sci, 2003, 68(2 Pt A), 263 - 6
Pycnoporus cinnabarinus laccases: an interesting tool for food or non-food applications; Georis J et al.; The effects of the addition of ferulic acid and ethanol in P . cinnabarinus ss3 culture medium in fermentor were compared in 15-L fermentor . In the presence of 30 g l(-1) ethanol, laccase activity (270,000 U/L1) was 3-fold higher as compared with ferulic acid-induced cultures, and 150-fold higher as compared with non-induced cultures, respectively . High-quality flax pulp was bleached in a totally-chlorine free (TCF) sequence using a laccase-mediator system constituted by laccase from Pycnoporus cinnabarinus and 1-hydroxybenzotriazole (HBT) as mediator . Up to 90% delignification and strong brightness increase were attained after the laccase-mediator treatment followed by H2O2 bleaching . This TCF sequence was further improved by applying H2O2 under pressurized O2 . In this way, up to 82% ISO brightness was obtained (compared with 37% in the initial pulp and 60% in the peroxide-bleached control) as well as very low kappa number . A positive evaluation of the laccase has been also performed in a food application . The colour of a tea-based beverage was significantly improved by incubating an infusion of green tea with the Pycnoporus laccase.

Commun Agric Appl Biol Sci, 2003, 68(2 Pt A), 191 - 4
Occurrence and origin of phosphine in landfill gas; Roels J et al.; A landfill (Hooge Maey, Flanders, Belgium) was subjected to an in-depth study in order to explain the origin of phosphine detected in high amounts in landfill gas during a previous study . The spatial and temporal variability of the phosphine concentration in landfill gas was assessed . Twenty four wells were monitored and differences in phosphine concentration up to one log unit were observed (3.2-32.4 microg/m3) . The phosphine concentration in each well was constant in time over a period of 4 months . No correlation was found between the phosphine concentration and methane, carbon dioxide, hydrogen sulphide, ethene or ethane concentration . In a series of laboratory tests, it was shown that phosphine was emitted during batch fermentation tests inoculated with landfill leachate when Fe0 or Al0 specimens were added . Conditions favouring corrosion of iron gave rise to higher emissions of phosphine . The phosphine concentration in the headspace of a batch test rose to 1.43 mg/m3 after 27 days of incubation . Weight loss of corroding steel coupons correlated with phosphine emission . Calculations showed that all phosphine emitted from the 0.005 km3 landfill (160 g/year) could be attributed to corrosion of metals . No evidence of de novo synthesis could be established

Commun Agric Appl Biol Sci, 2003, 68(2 Pt A), 167 - 70
Optimized biogas-fermentation by neural network control; Holubar P et al.; In this work several feed-forward back-propagation neural networks (FFBP) were trained in order to model, and subsequently control, methane production in anaerobic digesters . To produce data for the training of the neural nets, four anaerobic continuous stirred tank reactors (CSTR) were operated in steady-state conditions at organic loading rates (Br) of about 2 kg x m(-3) x d(-1) chemical oxygen demand (COD), and disturbed by pulse-like increase of the organic loading rate . For the pulses additional carbon sources were added to the basic feed (surplus- and primary sludge) to simulate cofermentation and to increase the COD . Measured parameters were: gas composition, methane production rate, volatile fatty acid concentration, pH, redox potential, volatile suspended solids and COD of feed and effluent . A hierarchical system of neural nets was developed and embedded in a Decision Support System (DSS) . A 3-3-1 FFBP simulated the pH with a regression coefficient of 0.82 . A 9-3-3 FFBP simulated the volatile fatty acid concentration in the sludge with a regression coefficient of 0.86 . And a 9-3-2 FFBP simulated the gas production and gas composition with a regression coefficient of 0.90 and 0.80 respectively . A lab-scale anaerobic CSTR controlled by this tool was able to maintain a methane concentration of about 60% at a rather high gas production rate of between 5 to 5.6 m3 x m(-3) x d(-1).

Commun Agric Appl Biol Sci, 2003, 68(2 Pt A), 109 - 14
Production of polyhydroxyalkanoates by a mixed culture in a sequencing batch reactor: the use of propionate as carbon source; Lemos PC et al.; In this work, sludge adapted to anaerobic/aerobic conditions, AN/AE, showing a high capacity of P accumulation, was submitted to aerobic dynamic substrate feeding (ADF) . The fermenter was operated as a Sequencing Batch Reactor, with propionate as carbon substrate . Propionate is an important waste product from several industrial processes that can be valued, using it as a precursor for hydroxyvalerate in PHA production . Under the operational conditions used, apart from 3-hydroxyvalerate as its major component, 3-hydroxy-2-methylvalerate, 2-hydroxyisovalerate and 4-oxovalerate were also produced . A second reactor operated under the same conditions was adapted for the use of acetate as carbon substrate . The global metabolism of the organisms involved on PHA production, utilizing acetate or propionate, was studied using in vivo 13C Nuclear Magnetic Resonance (NMR).

Curr Opin Clin Nutr Metab Care, 2004 Sep, 7(5), 563 - 7
The butyrate story: old wine in new bottles?
Scheppach W, Weiler F.
PURPOSE OF REVIEW: Short-chain fatty acids are important end products of bacterial carbohydrate fermentation in the colon . In particular, n-butyrate is thought to play a regulatory role in the maintenance of a physiological environment . Disturbances in the interplay between the microflora and the lining epithelium may lead to mucosal inflammation and promote carcinogenesis . The purpose of this article is to review the literature between March 2003 and February 2004 and to determine if recent studies have improved the understanding of butyrate effects in health and disease . RECENT FINDINGS: Preclinical studies (cell culture experiments, animal studies) using modern molecular biological tools (including cDNA arrays) have provided new insights into the action of butyrate on colonic epithelial, vascular endothelial and extracolonic cell types . The new information adds pieces of evidence to the assumption that butyrate may ameliorate colonic inflammation and may be chemopreventive in carcinogenesis . In contrast, new data from clinical studies have been limited in the review period . SUMMARY: In the era of molecular biology our understanding of subcellular processes that ultimately lead to inflammatory bowel disease or colorectal cancer has widened considerably . The new powerful technology of genomics and proteomics, however, raises new questions without easy answers . With this new information in mind, we will have to go back to human intervention trials to test the hypotheses generated in vitro . The preclinical data from the review period justify the need for carefully designed clinical trials to test the benefits derived from butyrate production.

Asia Pac J Clin Nutr . 2004;13(Suppl):S168.
Dietary and health characteristics of rural Korean farmer families with patients; Rhie SG et al.; Recent WTO and FTA led agricultural globalization has brought crisis to already impoverished Korean farmers . This study was proposed to assist in comparing health and dietary characteristics of farmer families with chronic disease patients to farmer families without chronic disease patients . For the study, 1870 families were selected from 9 rural Korean provinces . Trained evaluators interviewed farmer housewives to collect demographic, health behavior, and dietary relative information about family members . Statistical analyses were performed using SAS (ver 8.1) . Chi-square tests and General Linear Models were used . In general, patient family members were older than non-patient family members . For patient families, mean age was 70.4 for husbands and 64.3 for wives . For non-patient families, mean age was 64.2 for husbands and 57.3 for wives . Therefore we analyzed data after stratified subjects by age 65 . Patient families snacked less and "dined out" less than non-patient families . However, they consumed cookies more frequently, and milk and fruits less frequently, when compared to non-patient families . There were no significant differences in nutrient supplementation, food taboos, and/or instant food intake frequencies between patient families and non-patient families . Patient families made less homemade traditional fermented food (kimchi) than non-patient families . Sixty-two percent of patient family members complained about health problems such as shoulder stiffness, lumbago, numb limbs, dizziness, nocturia, breathlessness, sleeplessness, and abdominal fullness, whereas 52% of non-patient family members complained about health problems . Husband cigarette smoking was not significantly different among groups . However, smoking of wives was significantly higher in patient families . Alcohol consumption was also higher in patient families . In summary, it was determined that rural patient families had poorer dietary behavior and poorer health in general, when compared to non-patient families, and accordingly, diverse community-level health and nutrition support is suggested to solve farmers' health problems and to improve their quality of life.

Asia Pac J Clin Nutr . 2004;13(Suppl):S147.
Reduction of the postprandial glucose and insulin response in serum of healthy subjects by an arabinoxylan concentrate isolated from wheat starch plant process water; Zunft HJ et al.; Background: During wheat starch processing soluble constituents of flour such as proteins, pentosans, and minerals are dissolved in the process water and usually used for animal feeding . However, the pentosan fraction mainly consisting of arabinoxylans may be useful in human nutrition to lower the glycemic index of cereal products . Therefore, a new process for concentrating and purifying the pentosans has been developed . The resulting soluble dietary fiber concentrate has been studied for its ability to reduce the postprandial glucose and insulin response in healthy volunteers . Methods: After enzymatic, fermentative, mechanical treatment, cross flow ultrafiltration and spray-drying a product containing 60 % of arabinoxylan-enriched dietary fiber was obtained from wheat starch plant process water . The metabolic effect of the concentrate was investigated in two double-blind controlled studies with 11 (study 1) and 15 (study 2) healthy volunteers, respectively . At two mornings within a week each subject consumed isocaloric test meals (365 kcal) in a randomised order, one of them containing 6 grams of the arabinoxylan concentrate delivered via rolls . For measuring serum glucose and insulin blood samples were collected over the subsequent two hours . Results: The postprandial glucose response expressed as incremental area under curve was reduced by the arabinoxylan concentrate, in study 1 significantly (by 24 %, P<0.05), in study 2 in tendency (P = 0.051) . The postprandial insulin response showed significant reductions in both studies (by 9 % and 20%, P<0.05) . Conclusion: A spray-dried arabinoxylan concentrate administered via rolls in a breakfast is a soluble dietary fiber effective to lower the postprandial blood glucose and insulin response.

Asia Pac J Clin Nutr . 2004;13(Suppl):S120.
Iron bioavailability of some Cameroon traditional complementary foods; Kana Sop MM et al.; Iron "in vitro" bioavailability was assessed in main Cameroonian traditional complementary foods identified during enquiries amongst mothers with weaning babies up to 30 months by and "in vitro extrinsic tag method . Total iron levels were generally ranged between 4.68+/-0.5 (in fermented maize gruel) and 51.51+/-3.73 mg/ 100g dry matter (in maize meal with a vegetable, Corchorus olitorius based sauce) . Non haem iron values were high compared to haem iron values calculated by difference (between total and non hem iron) and ranged from 3.11+/-0.09 (in sweet potato with pear) to 47.64+/-4.68 mg/ 100g dry matter (in maize meal with a vegetable, Corchorus olitorius based sauce) . Dialysable iron values expressed in % of non hem iron ranged between 0.89+/-0.13 and 18.68+/-2.11 % (in mashed Irish potato with fish) were enhanced with lime juice and significantly reduced by legumes (beans, soy, and peanut), egg and egg yolk . An Irish potato-based diet was the best source of dialyzable Iron . Iron intakes were sufficient for most balanced diets to cover iron recommended daily intakes from 7 months of age.

Asia Pac J Clin Nutr . 2004;13(Suppl):S114.
The nutritional status of pregnant women in the Vaal Triangle, Gauteng, South Africa; Kesa H; Background: A study completed by H.Kesa (2001) formed part of a clinical intervention trial under controlled conditions to examine the iron status of the pregnant and lactating women in the Vaal Triangle . The main purpose of that study was to determine the dietary intake and iron status of the sample population (n=431) . Based on that study there is convincing evidence of poor dietary practices by pregnant women . According to Ramachandran (2002) low dietary intake in pregnancy will have adverse effects on the health and nutrition status of both the mother and her offspring . According to Norton (2002) there is strong epidemiological evidence of an association between maternal weight gain during pregnancy and low birth weight (LBW), especially in undernourished women . Aims: The main purpose of this study is to develop a cost effective, nutrient-dense food multimix for pregnant women in order to improve their nutritional status during pregnancy and to have healthier pregnancy outcomes . Methodology: According to the study by H.Kesa (2001) and Norton (2002) validated QFFQ's was used and statistically analysed . Trained fieldworkers conducted interviews with the help of food models to estimate portion size . Results: According to the study by H.Kesa (2001) the top ten items most often consumed by pregnant women were, in descending order: fresh milk, tea, coffee, cold drink, maize meal, fruit juice, bread rolls, magou (a fermented non-alcoholic drink), rice and sugar . Daily intakes (mean +/- SD) for pregnant women were: 8425.71 +/- 2279 kJ, 73.18 +/- 23 g protein, 62.29 +/- 23.7 g fat, 292.45 +/- 72.2 g carbohydrate, 9.74 +/- 3.8 mg iron . According to the blood samples 50% of pregnant women IDA . According to Norton (2002) majority of women with LBW babies are from developing countries where women begin pregnancy in a nutritionally disadvantaged state . Conclusions: Based on the results of the previous study, it is clear that food insecurity and malnutrition are evident in pregnant women in the Vaal Traingle . Implications: The development of a multimix will attempt to examine a cost- effective means to medium and long-term sustainable food based solutions to food and nutrition security of low-income, pregnant women in the Vaal Triangle in order to prevent malnutrition during pregnancy.

Asia Pac J Clin Nutr . 2004;13(Suppl):S23.
Strategies for preventing micronutrient deficiencies in developing countries; Gibson RS; Until recently, strategies for preventing micronutrient deficiencies have focussed on single micronutrients, principally iodine, iron, and vitamin A . The importance of concurrent micronutrient deficiencies in developing countries (DCs) is now recognized, their existence prompted by the often disappointing responses with single micronutrient supplementation programs . Latent deficiencies of other micronutrients can suppress the effect of a single micronutrient, when it is not the first limiting nutrient . The etiology of these multiple micronutrient deficiencies is multifactorial . Inadequate intakes and/or poor bioavailability, induced by predominately plant-based diets and low intakes of animal source foods, are major factors, although non-nutritional factors such as parasitic infections, genetic hemoglobinopathies, malaria, and infectious diseases, play a role . Co-existing micronutrient deficiencies result in impairments in growth, immune, and cognitive function, poor reproductive outcome, and increased morbidity and mortality . Clearly major health benefits could be achieved by choosing appropriate and cost-effective strategies that successfully alleviate concurrent micronutrient deficiencies in developing countries . Approaches include supplementation to those 'at risk', and food-based strategies involving fortification and dietary diversification/modification with minimal risk of antagonistic micronutrient interactions Of these, multi-micronutrient fortification of centrally processed staple foods or condiments is now feasible, but inappropriate in subsistence settings . Instead, for the latter, more sustainable approaches involve biofortification of plant-based staples, and promotion of small-livestock production, aquaculture, and consumption of animal source foods . In addition, household dietary strategies involving changes in food preparation and processing can be used to alter the content of micronutrient absorption modifiers in plant-based diets . Practical methods involve consumption of absorption enhancers, and use of germination, fermentation, and soaking to reduce the phytate content of cereal flours by enzyme-induced hydrolysis of phytate and/or passive diffusion of water soluble phytate . We have applied these dietary strategies among subsistence households in rural Malawi, and evaluated their impact on the dietary adequacy of weanlings and young children through knowledge and practices and interactive 24-h recalls, using a quasi-experimental design . In both groups, intervention diets were of higher dietary quality than controls, supplying significantly more animal source foods, especially soft-boned fish, but less phytic acid . Median intakes of energy, protein, calcium, available zinc, available iron (only for weanlings) were greater, and phytate:zinc molar ratios were lower in intervention compared to controls . In the children, intervention enhanced Z-scores for mid-upper-arm circumference and arm muscle area, but had no impact on weight or height gain . After controlling for baseline variables, mean hemoglobin was higher post-intervention, whereas incidence of anemia and common infections was lower in intervention compared to controls . To enhance effectiveness and sustainability, all micronutrient strategies should be integrated with ongoing national food, nutrition, and health education programs, and implemented using education and social-marketing techniques.

Clin Rheumatol, 2004 Aug, 23(4), 355 - 7 Epub 2004 Apr 16.
A case of anti-Jo1 myositis with pleural effusions and pericardial tamponade developing after exposure to a fermented Kombucha beverage; Derk CT et al.; The pathogenesis of the idiopathic inflammatory myopathies has been postulated to be an environmental trigger causing the expression of the disease in a genetically predisposed patient . We report a case of anti-Jo1 antibody-positive myositis which was associated with pleural effusions, pericardial effusion with tamponade, and 'mechanic's hands', probably related to the consumption of a fermented Kombucha beverage . Kombucha 'mushroom', a symbiosis of yeast and bacteria, is postulated to be the trigger for our patient's disease owing to the proximity of his symptoms to the consumption of the Kombucha beverage.

J Bacteriol, 2004 Aug, 186(16), 5418 - 26
Differential regulation of soluble and membrane-bound inorganic pyrophosphatases in the photosynthetic bacterium Rhodospirillum rubrum provides insights into pyrophosphate-based stress bioenergetics; Lopez-Marques RL et al.; Soluble and membrane-bound inorganic pyrophosphatases (sPPase and H(+)-PPase, respectively) of the purple nonsulfur bacterium Rhodospirillum rubrum are differentially regulated by environmental growth conditions . Both proteins and their transcripts were found in cells of anaerobic phototrophic batch cultures along all growth phases, although they displayed different time patterns . However, in aerobic cells that grow in the dark, which exhibited the highest growth rates, Northern and Western blot analyses as well as activity assays demonstrated high sPPase levels but no H(+)-PPase . It is noteworthy that H(+)-PPase is highly expressed in aerobic cells under acute salt stress (1 M NaCl) . H(+)-PPase was also present in anaerobic cells growing at reduced rates in the dark under either fermentative or anaerobic respiratory conditions . Since H(+)-PPase was detected not only under all anaerobic growth conditions but also under salt stress in aerobiosis, the corresponding gene is not invariably repressed by oxygen . Primer extension analyses showed that, under all anaerobic conditions tested, the R . rubrum H(+)-PPase gene utilizes two activator-dependent tandem promoters, one with an FNR-like sequence motif and the other with a RegA motif, whereas in aerobiosis under salt stress, the H(+)-PPase gene is transcribed from two further tandem promoters involving other transcription factors . These results demonstrate a tight transcriptional regulation of the H(+)-PPase gene, which appears to be induced in response to a variety of environmental conditions, all of which constrain cell energetics.

J Agric Food Chem, 2004 Aug 11, 52(16), 5117 - 23
Effect of the maceration technique on the relationships between anthocyanin composition and objective color of Syrah wines; Gomez-Miguez M et al.; The effects of two different vinification techniques, traditional fermentation and carbonic maceration, on the anthocyanin composition and color of young red wines, made with Syrah grapes grown in a warm climate, were compared . Tristimulus colorimetry was applied to study the color of wines during the vinification, and a high-performance liquid chromatography (HPLC) procedure was used for the analysis of anthocyanins . Carbonic maceration led to wines with lower anthocyanin content, mainly monoglucosides, and total phenols . This was related to lighter wines, less saturated, but more colorful (higher chroma C*ab values), and hues hab similar to those of the Syrah wines made by traditional vinification . Thus, the lightness L* had much more influence on the saturation s*uv of the wines obtained by carbonic maceration than the chroma (s*uv = C*uv/L*) . From a study of the color-composition relationships using linear and multiple regression, better relationships were found for the wines from traditional vinification, where the chromatic parameters L*, hab, and s*uv could be predicted from the 3-monoglucosides of delphinidin, petunidin, peonidin, and malvidin concentrations (R > 0.9) . However, a good prediction of the chroma C*ab from the anthocyanin composition was not possible . On the contrary, C*ab was the best predicted parameter from the anthocyanins monoglucosides (R > 0.9) in the carbonic maceration wines.

J Dairy Sci, 2004 May, 87(5), 1455 - 6
Short communication: effect of carbohydrate fermentation rate on estimates of mass fermented and milk response; Hall MB; Current prediction equations were used to evaluate the effects of rates of fermentation of fiber or starch in individual feeds on amounts of carbohydrate fermented ruminally and milk yield responses . The small predicted increases in carbohydrate fermented and milk response associated with doubling the rates of fermentation suggest that current prediction equations are relatively insensitive to changes in rate of fermentation.

J Dairy Sci, 2004 May, 87(5), 1446 - 54
Digestible energy values of diets with different fat supplements when fed to lactating dairy cows; Weiss WP et al.; A total collection digestion trial using high producing lactating cows (average milk yield = 40.7 kg/d) was conducted to measure the effect of different fat supplements on dietary digestible energy (DE) concentrations and to calculate the DE value of the supplements . A diet with no supplemental fat, 2 diets with 1.75 or 3.5% (dry basis) Ca salts of palm fatty acids (Ca-PFA), and 2 diets with 1.6 or 3.2% hydrogenated triacylglycerides from palm oil (HPO) were fed in a 5 x 5 Latin square experiment with 28-d periods . Concentrations of supplemental long-chain fatty acids in the diets were 1.7 and 3.4% for the 2 supplementation rates . Dry matter intake was reduced when cows were fed the high concentration of Ca-PFA, but cows fed Ca-PFA produced more milk than cows fed the control diet or diets with HPO . The type or amount of fat supplementation did not affect measures of rumen fermentation or in situ fiber digestibility . Digestibility of energy, dry matter, and organic matter were higher for diets with Ca-PFA than for diets with HPO, mainly because of increased fatty acid digestibility . The dietary concentration of DE was similar between the control diet and diets with HPO (2.97 Mcal/kg), but it increased as the concentration of Ca-PFA increased (3.04 and 3.16 Mcal/kg for low and high supplementation rates) . The calculated DE concentrations of the supplements averaged 7.3 and 3.1 Mcal/kg for Ca-PFA and HPO . The 2001 National Research Council dairy model accurately estimated DE concentrations in all diets (<1% difference).

J Dairy Sci, 2004 May, 87(5), 1413 - 24
Effects of fiber content and particle size of forage on the flow of microbial amino acids from continuous culture fermenters; Rodriguez-Prado M et al.; Eight dual-flow continuous culture fermenters (1320 mL) were used in two 10-d consecutive periods to study the effects of fiber content and particle size on rumen fermentation, nutrient flow, and the profile and flow of amino acids (AA) from microbial origin . Treatments were arranged in a 2 x 2 factorial; the main factors were fiber content (high fiber {HF} = 67% alfalfa hay, 33% concentrate; low fiber {LF} = 39% alfalfa hay, 61% concentrate) and forage stem particle size (ground not to pass a 3-mm sieve = large size {LS}; ground to pass a 1-mm sieve = small size {SS}) . On the last day of the experiment, liquid- (LAB) and solid- (SAB) associated bacteria were isolated from each fermenter for chemical analysis . Microbial N and AA flows were estimated using LAB or SAB composition data . Total volatile fatty acid concentration was higher for LF (122 mM) than for HF (102 mM) . The proportion of acetate was higher for HF (63.5%) than for LF (58.2%) . When SS was fed instead of LS, there was a reduction in the acetate proportion (63.4% vs 58.3%, respectively) and in the acetate to propionate ratio (3.13 vs . 2.41, respectively) . Bacterial N flow was higher, and dietary N flow was lower, in SS compared with LS when LAB were used for calculations . Efficiency of microbial protein synthesis (EMPS) was affected by particle size, when LAB were used for calculations, and by fiber content, when SAB were used for calculations . The AA profile of LAB differed from SAB in 4 of 16 AA . The fiber content and particle size had small effects on microbial AA profile . However, the bacterial AA flow was higher in 13 of 16 AA, when SAB instead of LAB were used for calculations . Estimates of bacterial N flow, EMPS, AA profile, and flows of microbial origin were dependent on the type of microbial population used for calculations . The representativeness of the microbial samples is essential for the accurate prediction of the AA flow from microbial origin.

J Dairy Sci, 2004 May, 87(5), 1360 - 71
Effect of inhibitor concentration and end-product accumulation on estimates of ruminal in vitro protein degradation; Broderick GA et al.; Effects of varying the concentrations of hydrazine sulfate (HS) and chloramphenicol (CAP), inhibitors of microbial-N uptake and protein synthesis, on rates of protein degradation estimated from net appearance of NH3 and total amino acids (TAA) were studied in a ruminal in vitro fermentation system . Without inhibitors, recoveries of N added as NH3 and TAA were 4 and 6% after 4-h incubations, and apparent degradation rates estimated from release of NH3 and TAA for casein, solvent soybean meal (SSBM), and expeller soybean meal (ESBM) approached 0 . Increasing inhibitor concentrations from the standard amounts of 1 mM HS plus 30 mg of CAP/L to 2 mM HS plus 90 mg of CAP/ L gave rise to numerically greater N recoveries and degradation rates, but these differences were not statistically significant . Compared with the standard inhibitor concentrations, use of 2 mM HS, without CAP, yielded similar recoveries and rates, but 30 or 90 mg of CAP/L, without HS, was not satisfactory . Versus that with 1 mM HS plus 30 mg of CAP/L, media containing 2 mM HS plus 90 mg of CAP/L gave increased TAA recoveries and higher rates for casein, but not SSBM, in the presence of added starch . Faster degradation rates were obtained for casein, but slower rates for SSBM and ESBM, in Sweden versus Wisconsin using inocula from cows fed different diets but with similar CP and energy contents . Differences in microbial catabolism of peptides may account for differences in degradation rates observed between Sweden and Wisconsin . Adding NH3 plus free and peptide-bound amino acids to the inoculum reduced apparent degradation rates, possibly via end-product inhibition . Analysis of data from multiple time-point incubations indicated that casein degradation followed simple, first-order kinetics, while a biexponential model fitted degradation patterns for both SSBM and ESBM.

Emerg Radiol, 2003 Dec, 10(3), 163 - 5 Epub 2003 Aug 23.
Ruptured splenic abscess: a cause of pneumoperitoneum in a patient with AIDS; Ishigami K et al.; We encountered a case of pneumoperitoneum caused by a gas-forming splenic abscess in a patient with acquired immunodeficiency syndrome (AIDS) . Plain abdominal films and computed tomography demonstrated a large amount of free air . Pneumoperitoneum was eventually shown to represent gas liberated from fermentation by gas-forming organisms within the splenic abscess . Gas-containing necrotic tissue from the ruptured spleen mimicked the spillage of feces from colon perforation . The authors emphasize that a ruptured abscess should be included in the differential diagnosis of acute abdomen in an immunocompromised patient.

Emerg Radiol, 2004 Apr, 10(5), 279 - 81 Epub 2004 Mar 17.
Intravascular gas in the transplanted kidney: a sign of extensive graft necrosis; Ishigami K et al.; We encountered a case of transplanted kidney necrosis, with computed tomography (CT) demonstrating multiple areas of intravascular gas within the allograft . The intravascular gas represented air emboli from gas liberated from fermentation by gas-forming organisms in a perinephric abscess . Arterial bleeding accelerated by the wound infection and the resultant large perinephric hematoma caused renal infarction . Gas-forming infection of transplanted organs is associated with a poor graft outcome, which can present as a fulminant clinical course . Intravascular gas should be distinguished from collecting system gas because the former could represent extensive necrosis of the transplanted kidney.

Appl Microbiol Biotechnol . 2004 Jul 30; {Epub ahead of print}
Astaxanthin hyperproduction by Phaffia rhodozyma (now Xanthophyllomyces dendrorhous) with raw coconut milk as sole source of energy; Dominguez-Bocanegra AR et al.; Natural carbon sources, such as those present in cane sugar molasses and grape juice, promote the synthesis of astaxanthin in different Phaffia rhodozyma yeasts . One of these, coconut milk, has a very rich nutrient composition . The aim of this work was to investigate the utility of coconut milk as sole source of energy for astaxanthin pigment production by P . rhodozyma strains . Currently, coconut pulp is widely used in industrial processes in Mexico for the production of shampoos, candies, food, etc . However, coconut milk is a waste product . We show that coconut milk enhances astaxanthin production . The fermentation yielded 850 microg/g yeast with the NRRL-10921 wild-type strain and 1,850 microg/g yeast with the mutated R1 strain . Production was better than reported results employing other natural carbon sources.

Appl Microbiol Biotechnol . 2004 Jul 28; {Epub ahead of print}
The effect of decreasing oxygen feed rates on growth and metabolism of Torulaspora delbrueckii; Hanl L et al.; The effect of decreasing oxygen feed rates on the growth and metabolism of Torulaspora delbrueckii and Saccharomyces cerevisiae in chemostat cultures was investigated . The biosynthetic oxygen requirement, i.show $132#e . the minimum specific oxygen consumption rate required for steady-state growth at a dilution rate of 0.10 h(-1), of T . delbrueckii was quantified to be less than 0.1 mmol O(2) g(-1) h(-1) . Under strict anaerobiosis, washout of T . delbrueckii occurred, whereas for S . cerevisiae it did not . Under oxygen-limited conditions, the increase in fermentative ability of T . delbrueckii with diminishing oxygen supply was less pronounced than that of S . cerevisiae . These results indicate that T . delbrueckii was more disturbed in its energy balance than S . cerevisiae under strict anaerobiosis, and they may explain why T . delbrueckii exhibits poorer growth than S . cerevisiae under this condition.

Appl Microbiol Biotechnol . 2004 Jul 30; {Epub ahead of print}
d-Hexosaminate production by oxidative fermentation; Moonmangmee D et al.; Microbial production of d-hexosaminate was examined by means of oxidative fermentation with acetic acid bacteria . In most strains of acetic acid bacteria, membrane-bound d-glucosamine dehydrogenase (synonymous with an alternative d-glucose dehydrogenase distinct from quinoprotein d-glucose dehydrogenase) oxidized d-hexosamines to the corresponding d-hexosaminates in a stoichiometric manner . Conversion of d-hexosamines to the corresponding d-hexosaminates was observed with growing cells of acetic acid bacteria, and d-hexosaminate was stably accumulated in the culture medium even though d-hexosamine was exhausted . Since the enzyme responsible is located on the outer surface of the cytoplasmic membrane, and the enzyme activity is linked to the respiratory chain of the organisms, resting cells, dried cells, and immobilized cells of acetic acid bacteria were effective catalysts for d-hexosaminate production . d-Mannosaminate and d-galactosaminate were also prepared for the first time by means of oxidative fermentation, and three different d-hexosaminates were isolated from unreacted substrate by a chromatographic separation . In this paper, d-hexosaminate production by oxidative fermentation carried out mainly with Gluconobacter frateurii IFO 3264 is exemplified as a typical example.

Biotechnol Lett, 2004 Aug, 26(15), 1241 - 5
Production of sophorolipids by Candida bombicola grown on soy molasses as substrate; Solaiman DK et al.; Sophorolipids (SLs) were produced from Candida bombicola using soy molasses and oleic acid as co-substrates . The purified SLs were obtained at 21 g l(-1) and were 97% in lactone form . The major SL constituent (81% relative abundance) of the product mixture contains an oleoyl chain . The surface properties of the SLs obtained from the soy molasses/oleic acid fermentation had minimum surface-tension values of 37 mN m(-1) (pH 6) and 38 mN m(-1) (pH 9), and critical micelle concentration values of 6 mg l(-1) (pH 6) and 13 mg l(-1) (pH 9).

Microbiology, 2004 Aug, 150(Pt 8), 2761 - 74
Heterologous production of the antifungal polyketide antibiotic soraphen A of Sorangium cellulosum So ce26 in Streptomyces lividans; Zirkle R et al.; The antifungal polyketide soraphen A is produced by the myxobacterium Sorangium cellulosum So ce26 . The slow growth, swarming motility and general intransigence of the strain for genetic manipulations make industrial strain development, large-scale fermentation and combinatorial biosynthetic manipulation of the soraphen producer very challenging . To provide a better host for soraphen A production and molecular engineering, the biosynthetic gene cluster for this secondary metabolite was integrated into the chromosome of Streptomyces lividans ZX7 . The upstream border of the gene cluster in Sor . cellulosum was defined by disrupting sorC, which is proposed to take part in the biosynthesis of methoxymalonyl-coenzyme A, to yield a Sor . cellulosum strain with abolished soraphen A production . Insertional inactivation of orf2 further upstream of sorC had no effect on soraphen A production . The genes sorR, C, D, F and E thus implicated in soraphen biosynthesis were then introduced into an engineered Str . lividans strain that carried the polyketide synthase genes sorA and sorB, and the methyltransferase gene sorM integrated into its chromosome . A benzoate-coenzyme A ligase from Rhodopseudomonas palustris was also included in some constructs . Fermentations with the engineered Str . lividans strains in the presence of benzoate and/or cinnamate yielded soraphen A . Further feeding experiments were used to delineate the biosynthesis of the benzoyl-coenzyme A starter unit of soraphen A in the heterologous host.

J AOAC Int, 2004 May-Jun, 87(3), 792 - 6
Physiological functions of resistant proteins: proteins and peptides regulating large bowel fermentation of indigestible polysaccharide; Morita T et al.; Animal studies have shown conclusively that feeding of resistant starch (RS) increases production of large bowel total short-chain fatty acids (SCFAs) . However, fermentation products of RS may be affected considerably by other dietary ingredients . In rats fed a 20% high-amylose cornstarch (HAS) with casein as the sole protein source, greater cecal SCFAs production was observed compared with that in rats fed a regular cornstarch diet . However, with this diet, the cecal succinate production was also very high . In contrast, when rice or potato protein with lower digestibility was used in place of casein, cecal succinate production decreased with a concomitant increase in butyrate . These observations suggest that nondigested protein, namely resistant protein, might play a role in correcting an imbalance in the ratio of carbohydrate and nitrogen as fermentative substrates for cecal bacteria and in promoting butyrate production . Epidemiological and biochemical data indicate a possible linkage between the fermentation products of starch (butyrate in particular) and the prevention of colorectal cancer as well as ulcerative colits . Accordingly, a fermentation strategy of RS favoring SCFA production should be established to elucidate the potentially beneficial effects of SCFAs on large bowel physiology.

J AOAC Int, 2004 May-Jun, 87(3), 761 - 8
Resistant starch: metabolic effects and potential health benefits; Higgins JA; Although there is strong evidence that the amount and type of fat in the diet can have dramatic effects on metabolism, the case for carbohydrate subtypes influencing metabolic parameters is emerging . By definition, resistant starch (RS) is any starch that is not digested in the small intestine but passes to the large bowel . Here, RS is a good substrate for fermentation which gives rise to an increase in short-chain fatty acid production . The differing rates of absorption between RS and digestible starch are thought to denote their differential metabolic responses . RS intake is associated with several changes in metabolism which may confer some health benefits . RS intake seems to decrease postprandial glycemic and insulinemic responses, lower plasma cholesterol and triglyceride concentrations, improve whole body insulin sensitivity, increase satiety, and reduce fat storage . These properties make RS an attractive dietary target for the prevention of diseases associated with dyslipidemia and insulin resistance as well as the development of weight loss diets and dietary therapies for the treatment of Type 2 diabetes and coronary heart disease . This review analyzes the body of literature examining the metabolic effects of RS consumption and discusses possible mechanisms whereby increased short-chain fatty acid production in the bowel could account for some of these effects . The effects of RS in the large bowel per se are the topic of other reviews and are not addressed in this paper.

J AOAC Int, 2004 May-Jun, 87(3), 749 - 55
Physiological aspects of resistant starch and in vivo measurements; Champ MM; Resistant starch (RS) is the sum of starch and products of starch degradation not absorbed in the small intestine of healthy individuals . There are a number of RS with different characteristics which may have a different fate in the colon . As a consequence, all RS should not be considered equivalent as far as physiological properties are concerned; indeed, they may have a different impact on colonic health . This statement may explain part of the apparent contradictions in the literature on RS and cancer or inflammatory disease prevention . RS is fermented in the large intestine into short-chain fatty acids and, among those, butyrate, which is recognized as the main nutrient of the colonocyte . This fermentation pattern seems to be responsible for most of the effects of RS on colonic health . Another important property is linked to its ability to lower colonic pH, which is usually considered as beneficial for mineral biovailability in the colon or cancer prevention . Due to their fate in the digestive tract, RS materials do not seem to have any significant impact on glucose absorption or metabolism . On the contrary, they may have a hypocholesterolemic effect, but available information is contradictory.

J AOAC Int, 2004 May-Jun, 87(3), 621 - 31
Determination of the 13C/12C ratio of ethanol derived from fruit juices and maple syrup by isotope ratio mass spectrometry: collaborative study; Jamin E et al.; A collaborative study of the carbon-13 isotope ratio mass spectrometry (13C-IRMS) method based on fermentation ethanol for detecting some sugar additions in fruit juices and maple syrup is reported . This method is complementary to the site-specific natural isotope fractionation by nuclear magnetic resonance (SNIF-NMR) method for detecting added beet sugar in the same products (AOAC Official Methods 995.17 and 2000.19), and uses the same initial steps to recover pure ethanol . The fruit juices or maple syrups are completely fermented with yeast, and the alcohol is distilled with a quantitative yield (>96%) . The carbon-13 deviation (delta13C) of ethanol is then determined by IRMS . This parameter becomes less negative when exogenous sugar derived from plants exhibiting a C4 metabolism (e.g., corn or cane) is added to a juice obtained from plants exhibiting a C3 metabolism (most common fruits except pineapple) or to maple syrup . Conversely, the delta13C of ethanol becomes more negative when exogenous sugar derived from C3 plants (e.g., beet, wheat, rice) is added to pineapple products . Twelve laboratories analyzed 2 materials (orange juice and pure cane sugar) in blind duplicate and 4 sugar-adulterated materials (orange juice, maple syrup, pineapple juice, and apple juice) as Youden pairs . The precision of that method for measuring delta13C was similar to that of other methods applied to wine ethanol or extracted sugars in juices . The within-laboratory (Sr) values ranged from 0.06 to 0.16%o (r = 0.17 to 0.46 percent per thousand), and the among-laboratories (SR) values ranged from 0.17 to 0.26 percent per thousand (R = 0.49 to 0.73 percent per thousand) . The Study Directors recommend that the method be adopted as First Action by AOAC INTERNATIONAL.

Cell Death Differ, 2004 Nov, 11(11), 1204 - 12
Mycoplasma fermentans inhibits tumor necrosis factor alpha-induced apoptosis in the human myelomonocytic U937 cell line; Gerlic M et al.; Mycoplasma fermentans (M . fermentans) was shown to be involved in the alteration of several eukaryotic cell functions (i.e . cytokine production, gene expression), and was suggested as a causative agent in arthritic diseases involving impaired apoptosis . We investigated whether M . fermentans has a pathogenic potential by affecting tumor necrosis factor (TNF)alpha-induced apoptosis in the human myelomonocytic U937 cell line . A significant reduction in the TNFalpha-induced apoptosis (approximately 60%) was demonstrated upon either infection with live M . fermentans or by stimulation with non-live M . fermentans . To investigate the mechanism of M . fermentans antiapoptotic effect, the reduction of mitochondrial transmembrane potential (DeltaPsim) and the protease activity of caspase-8 were measured . In the infected cells, the reduction of DeltaPsim was inhibited (approximately 75%), and an approximately 60% reduction of caspase-8 activity was measured . In conclusion, M . fermentans significantly inhibits TNFalpha-induced apoptosis in U937 cells, and its effect is upstream of the mitochondria and upstream of caspase-8.

J Nutr, 2004 Aug, 134(8), 1942 - 7
Ingestion of guar gum hydrolysate, a soluble and fermentable nondigestible saccharide, improves glucose intolerance and prevents hypertriglyceridemia in rats fed fructose; Suzuki T et al.; Fructose feeding provides a dietary model of insulin resistance accompanied by hypertriglyceridemia . We examined the effects of guar gum hydrolysate (GGH), a soluble and fermentable nondigestible saccharide with low viscosity, on glucose intolerance and hypertriglyceridemia in rats fed high-fructose diets . Rats were fed either a dextrin-based or a fructose-based diet with or without GGH (75 g/kg) for 30 d . Oral glucose tolerance tests (OGTTs) were performed 0, 14, and 28 d after feeding . High-fructose feeding negatively affected glucose tolerance on d 14 and 28 . The addition of GGH to the diets improved glucose intolerance on d 28 . Fructose feeding induced hyperinsulinemia after an oral glucose load; this was also improved by GGH on d 28 . The glycogen concentration in the gastrocnemius muscles of rats was lowered by dietary fructose, and GGH supplementation abolished this decrease . Triglycerides in the plasma and livers of rats fed fructose diets were elevated, and the increases were ameliorated by supplemental GGH . Regardless of the type of carbohydrate, GGH enlarged the cecum and increased the cecal SCFA pools . In conclusion, supplemental feeding of GGH to rats improved the glucose intolerance and hypertriglyceridemia induced by a high-fructose diet . Possible mediators of these beneficial effects of GGH are the SCFAs produced by microbial fermentation of GGH in the large intestine.

Yeast, 2004 Jul 15, 21(9), 747 - 57
HXT5 expression is under control of STRE and HAP elements in the HXT5 promoter; Verwaal R et al.; Hexose transporter (Hxt) proteins transport glucose across the plasma membrane in the yeast Saccharomyces cerevisiae . Recently, we have shown that expression of HXT5 is regulated by the growth rate of the cells . Because gene expression is regulated by binding of specific transcription factors to regulatory elements in the promoters of genes, the presence of putative regulatory elements in the promoter of HXT5 was determined by computer-assisted analysis . This revealed the presence of two putative stress-responsive elements (STREs), one putative post-diauxic shift (PDS) element and two putative Hap2/3/4/5p (HAP) complex binding elements . The involvement of these elements was studied by using mutations in a HXT5 promoter-LacZ fusion construct . Growth during various conditions that result in low growth rates of yeast cells revealed that the STRE most proximal to the translation initiation site seemed to be involved in particular in regulation of HXT5 expression during growth at decreased growth rates . In addition, the HAP elements seemed to be required during growth on non-fermentable carbon sources . The PDS element and, to a lesser extent, the other STRE showed particular involvement in regulation of HXT5 expression during growth on ethanol . Finally, it was shown that the PKA pathway, which is known to be involved in expression of STRE-regulated genes, was also involved in regulation of HXT5 expression . A possible mechanism by which expression of HXT5 could be regulated by the transcriptional regulatory elements in the promoter is discussed .

Hum Mol Genet, 2004 Oct 1, 13(19), 2279 - 88 Epub 2004 Jul 28.
The expression of human mitochondrial ferritin rescues respiratory function in frataxin-deficient yeast; Campanella A et al.; Mitochondrial ferritin (MtF) is structurally and functionally similar to the cytosolic ferritins, molecules designed to store and detoxify cellular iron . MtF expression in human and mouse is restricted to the testis and few tissues, and it is abundant in the erythroblasts of patients with sideroblastic anemia, where it is thought to protect the mitochondria from the damage caused by iron loading . Mitochondria iron overload occurs also in cells deficient in frataxin, a mitochondrial protein involved in iron handling and implicated in Friedreich ataxia . We expressed human MtF in frataxin-deficient yeast cells, a well-characterized model of mitochondrial iron overload and oxidative damage . The human MtF precursor was efficiently imported by yeast mitochondria and processed to functional ferritin that actively sequestered iron in the organelle . MtF expression rescued the respiratory deficiency caused by the loss of frataxin protecting the activity of iron-sulfur enzymes and enabling frataxin-deficient cells to grow on non-fermentable carbon sources . Furthermore, MtF expression prevented the development of mitochondrial iron overload, preserved mitochondrial DNA integrity and increased cell resistance to H2O2 . The data show that MtF can substitute for most frataxin functions in yeast, suggesting that frataxin is directly involved in mitochondrial iron-binding and detoxification.

J Appl Microbiol, 2004, 97(3), 647 - 55
The role of indigenous yeasts in traditional Irish cider fermentations; Morrissey WF et al.; AIMS: To study the role of the indigenous yeast flora in traditional Irish cider fermentations . METHODS AND RESULTS: Wallerstein laboratory nutrient agar supplemented with biotin, ferric ammonium citrate, calcium carbonate and ethanol was employed together with PCR-restriction fragment length polymorphism analysis of the region spanning the internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene in the identification of indigenous yeasts at the species level, from traditional Irish cider fermentations . By combining the molecular approach and the presumptive media it was possible to distinguish between a large number of yeast species, and to track them within cider fermentations . The Irish cider fermentation process can be divided into three sequential phases based on the predominant yeast type present . Kloeckera/Hanseniaspora uvarum type yeasts predominate in the initial 'fruit yeast phase' . Thereafter Saccharomyces cerevisiae type yeast dominate in the 'fermentation phase', where the alcoholic fermentation takes place . Finally the 'maturation phase' which follows, is dominated by Dekkera and Brettanomyces type yeasts . H . uvarum type yeast were found to have originated from the fruit . Brettanomyces type yeast could be traced back to the press house, and also to the fruit . The press house was identified as having high levels of S . cerevisiae type yeast . A strong link was noted between the temperature profile of the cider fermentations, which ranged from 22 to 35 degrees C and the yeast strain population dynamics . CONCLUSIONS: Many different indigenous yeast species were identified . The mycology of Irish cider fermentations appears to be very similar to that which has previously been reported in the wine industry . SIGNIFICANCE AND IMPACT OF THE STUDY: This study has allowed us to gain a better understanding of the role of indigenous yeast species in 'Natural' Irish cider fermentations .

J Appl Microbiol, 2004, 97(3), 540 - 5
Growth and fermentation patterns of Saccharomyces cerevisiae under different ammonium concentrations and its implications in winemaking industry; Mendes-Ferreira A et al.; AIMS: To study the effects of assimilable nitrogen concentration on growth profile and on fermentation kinetics of Saccharomyces cerevisiae . METHODS AND RESULTS: Saccharomyces cerevisiae was grown in batch in a defined medium with glucose (200 g l(-1)) as the only carbon and energy source, and nitrogen supplied as ammonium sulphate or phosphate forms under different concentrations . The initial nitrogen concentration in the media had no effect on specific growth rates of the yeast strain PYCC 4072 . However, fermentation rate and the time required for completion of the alcoholic fermentation were strongly dependent on nitrogen availability . At the stationary phase, the addition of ammonium was effective in increasing cell population, fermentation rate and ethanol . CONCLUSIONS: The yeast strain required a minimum of 267 mg N l(-1) to attain complete dryness of media, within the time considered for the experiments . Lower levels were enough to support growth, although leading to sluggish or stuck fermentation . SIGNIFICANCE AND IMPACT OF THE STUDY: The findings reported here contribute to elucidate the role of nitrogen on growth and fermentation performance of wine yeast . This information might be useful to the wine industry where excessive addition of nitrogen to prevent sluggish or stuck fermentation might have a negative impact on wine stability and quality .

J Environ Pathol Toxicol Oncol, 1999, 18(2), 141 - 6
Ileal effluent as a fermentation substrate: implications for butyrate production in the colon; Robertson JA et al.; Fermentation of fiber can lead to an enhanced production of short chain fatty acids (SCFA) and, hence, contribute to the proposed anticarcinogenic properties of butyrate in the colon . The fermentation of fiber isolates and the corresponding ileal effluents has been compared under in vitro conditions . Yield of SCFA per gram of substrate fermented was similar for isolates and fiber-enriched effluents (approximately 4.9 mmol/g) and it could be inferred that nonfiber components of effluent also generated SCFA . Butyrate production was highest for glucan-based polymers (approximately 30% total SCFA) and, from the measured acidogenic profile, production of SCFA will occur mainly in the proximal colon . The buffering capacity of ileal effluents during fermentation restrict the potential for a reduction in pH during acidogenesis compared to fiber isolates . This buffering capacity could limit the bioavailability of butyrate in the colon and, hence, the ability to satisfy the proposed antineoplastic properties of butyrate in the colon.

Biotechnol Bioeng, 2004 Aug 5, 87(3), 259 - 73
Bioseparations; Lightfoot EN et al.; Here we review key applications of separation technology in applied biology . We first sketch out the field as a whole, but then narrow our scope to the processing of fermentation products, particularly to high-value biologicals such as proteins and nucleotides . We go on to provide a qualitative overview describing the importance and general nature of this large field, major trends, and the strategies that have proven most fruitful in evolving effective separation and purification processes . We then give a detailed description of individual separations equipment and the principles governing their operation . We concentrate throughout on making the available literature accessible to the reader; we provide what is hoped to be a representative set of basic references . However, these references, in turn, include some that suggest promising new developments as well as a number of more specialized reviews . We hope that our overall result provides the reader with access to the most relevant literature.

J Econ Entomol, 2004 Jun, 97(3), 757 - 63
Evaluation of methods for extracting Xylella fastidiosa DNA from the glassy-winged sharpshooter; Bextine B et al.; The recent spread of the plant pathogenic bacterium Xylclla fastidiosa Wells et al . by an invasive vector species, Homalodisca coagulata Say, in southern California has resulted in new epidemics of Pierce's disease of grapevine . Our goal is to develop an efficient method to detect low titers of X . fastidiosa in H . coagulata that is amenable to large sample sizes for epidemiological studies . Detection of the plant pathogenic bacterium X . fastidiosa in its insect vector is complicated by low titers of bacteria, difficulty in releasing it from the insect mouthparts and foregut, and the presence of substances in the insect that inhibit polymerase chain reaction (PCr) . To select the optimal protocol for DNA extraction to be used with PCR, we compared three standard methods and 11 commercially available kits for relative efficiency of X . fastidiosa DNA extraction in the presence of insect tissue . All of the protocols tested were proficient at extracting DNA from pure bacterial culture (1 x 10(5) cells), and all but one protocol successfully extracted sufficient bacterial DNA in the presence of insect tissue . Three DNA extraction techniques, immunomagnetic separation, the DNeasy Tissue kit (Qiagen, Hercules, CA), and Genomic DNA Purification kit (Fermentus, Hanover, MD), were compared more closely using a dilution series of X . fastidiosa (5000-0 cells) with and without insect tissue present . The DNeasy Tissue kit was the best kit tested, allowing detection of 5 x 10(3) X . fastidiosa cells with an insect head background.

Curr Genet, 2004 Sep, 46(3), 140 - 6 Epub 2004 Jul 27.
Expression studies of GUP1 and GUP2, genes involved in glycerol active transport in Saccharomyces cerevisiae, using semi-quantitative RT-PCR; Oliveira R et al.; Glycerol active uptake in Saccharomyces cerevisiae, characterised physiologically as a proton symport, was previously described as repressed by glucose, induced by growth on non-fermentable carbon sources and unresponsive to growth under salt stress . GUP1 and GUP2 were identified and characterised as genes involved in glycerol active uptake . Using semi-quantitative RT-PCR, GUP1 and GUP2 transcription was measured . Unlike active transport activity determined previously, this was shown to be constitutive and not affected by either glucose repression or growth under salt stress . Furthermore, transcription of GUP1 and GUP2 was not affected in the gpd1gpd2 mutant strain grown under salt stress in the presence of small amounts of glycerol, in which case a very high Vmax of glycerol uptake was reported . Intracellular compounds were determined . Glycerol, acetate and trehalose were found to be the major compounds accumulated . Surprisingly, the gpd1gpd2 mutant was found to produce significant amounts of glycerol . Yet, the results provide no evidence for a correlation between the amount of each compound and the glycerol transport activity in any of the strains .

Biosci Biotechnol Biochem, 2004 Jul, 68(7), 1588 - 90
8-Hydroxydaidzein, an aldose reductase inhibitor from okara fermented with Aspergillus sp . HK-388; Fujita T et al.; The aldose reductase (AR) inhibitor, 8-hydroxydaidzein, was isolated and identified from a methanolic extract of okara (soybean pulp) fermented with the fungal strain, Aspergillus sp . HK-388 . 8-Hydroxydaidzein showed non-competitive inhibition of human recombinant AR with respect to DL-glyceraldehyde, its Ki value being evaluated as 7.0 microM.

Physiol Behav, 2004 Sep 15, 82(2-3), 357 - 68
Anxiety and aggression associated with the fermentation of carbohydrates in the hindgut of rats; Hanstock TL et al.; Lactic acid accumulation in the caecum and colon resulting from the fermentation of carbohydrates can lead to deleterious effects in ruminant and monogastric animals, including humans . In the present study, we examined the behavioural effects of two types of commonly consumed foods: soluble and fermentable carbohydrates (FCs) . Thirty-six male Wistar rats were fed either a commercial rat and mouse chow, a soluble carbohydrate (SC)-based diet or an FC-based diet . Social interaction, anxiety, aggression and locomotor activity were examined by employing a social interaction test and a light/dark emergence test, while physical parameters of hindgut fermentation were examined after sacrifice, either 3 or 21 h after feeding . Results showed that anxiety (spending less time in the light compartment during the light/dark emergence test) and aggression (increased fighting during the social interaction test) were increased following raised concentrations of fermentation end products, such as lactic acid and volatile fatty acids (VFAs) in the caecum of rats . These associations occurred regardless of dopamine and 5-HT concentrations in the prefrontal cortex (PFC) and provide evidence supporting a general effect of FCs on behaviour . Possible mechanisms of action along with similarities between a rat and human model of acidosis are discussed.

Chemosphere, 2004 Sep, 56(11), 1121 - 7
Photolysis of spinosyns in seawater, stream water and various aqueous solutions; Liu S et al.; Spinosad, a reduced-risk insecticide, contains primarily two active compounds, spinosyns A and D that are fermentation products of bacterium Saccharopolyspora spinosa . It is currently used to control fruit flies in Hawaii, USA . In this study, we investigated photodegradation of spinosyns A and D, respectively, in seawater, stream, tap and distilled-deionized waters under various light sources . Photodegradation of the two chemicals was also studied in various aqueous solutions prepared with phosphate buffer at different pH or chemical sensitizers . Two major photolytic products from spinosyn A were detected as spinosyn B and hydroxylated spinosyn A . Spinosyn D was similarly hydroxylated and N-demethylated . Spinosyns A and D were photodegraded rapidly under sunlight in Hawaii, USA . The half-life of spinosyns A and D in stream water was 1.1 and 1.0 h, respectively, and was a half of that in distilled-deionized water, 2.2 and 2.0 h, respectively . Photodegradation of spinosyns A and D followed an order of increasing rate constants in distilled-deionized, seawater, stream and tap water under 300 nm artificial light, and was enhanced approximately 8- and 17-fold, respectively, in acetone-sensitized solution as compared to in distilled-deionized water . Photolysis rates of spinosyns A and D in isopropanol- or humic acid-fortified water did not differ much as compared with those accordingly in distilled-deionized water . Spinosyns A and D photodegraded slower in acidic aqueous solution than in basic aqueous solution.

Artif Cells Blood Substit Immobil Biotechnol, 2004 May, 32(2), 275 - 91
Encapsulation of brewers yeast in chitosan coated carrageenan microspheres by emulsification/thermal gelation; Raymond MC et al.; Brewers yeast was encapsulated in kappa-carrageenan microspheres using an emulsification-thermal gelation approach . Due to heat sensitivity of the yeast at temperatures in excess of 36 degrees C, mixtures of low and high gelation temperature carrageenans were tested to obtain a blend yielding a gelation temperature under 40 degrees C . A 20:80 dispersion of 2% carrageenan sol containing cells, in warm canola oil, produced microspheres upon cooling, with a mean diameter of 450 microm and narrow size dispersion (span of 1.2) . Application of a chitosan membrane coat to minimize cell release, increased the mean microsphere diameter to 700 microm, due to the coat thickness and swelling of the microspheres . This diameter was designed so as to minimize mass transfer limitations . Batch fermentations were carried out in a 3 L reactor on a commercial wort medium . Cell loading was 10(7) cells mL(-1) microspheres, and cell "burst" release was observed upon inoculation into fresh medium, whether microspheres were coated or not . The kinetics of intra- and extracapsular cell growth were determined . Increased concentrations of extracapsular free cells could be accounted for by growth in the wort medium, and by ongoing release from the gel microspheres, whether coated or not . Cell release from chitosan-coated carrageenan microspheres was less than that from uncoated microspheres, likely due to retention by the membrane coat . Growth kinetics and alpha-amino nitrogen consumption of encapsulated yeast were higher than that of free cells, and differences in alcohol and ester profiles were also observed, likely due to modified metabolism of the encapsulated yeast.

Methods Mol Biol, 2004, 267, 287 - 96
Multiple gene expression by chromosomal integration and CRE-loxP-mediated marker recycling in Saccharomyces cerevisiae; Johansson B et al.; Multiple gene expression can be introduced in a yeast strain with using only two markers by means of the two new vectors described, the expression vector pB3 PGK and the CRE recombinase vector pCRE3 . The pB3 PGK has a zeocin-selectable marker flanked by loxP sequences and an expression cassette consisting of the strong PGK1 promoter and the GCY1 terminator . The gene of interest (YFG1) is cloned between the promoter and terminator of pB3 PGK . The pB3 PGK-YFG1 is integrated into the genome by a single restriction cut within the YFG1 gene and integrated in the YFG1 locus . The strain is further transformed with the pCRE3 vector . The CRE recombinase expressed from this vector removes the zeocin marker and makes it possible to use the pB3 PGK vector over again in the same strain after curing of the pCRE3 vector . The 2 micro -based pCRE3 carries the aureobasidin A, zeocin and URA3 markers . pCRE3 is easily cured by growth in nonselective medium without active counterselection . The screening for loss of the chromosomal zeocin marker, as well as curing of the pCRE3 vector, is done in one step, by scoring zeocin sensitivity . This can be done because the zeocin marker is present in both the pB3 PGK and pCRE3 . The S . cerevisiae pentose phosphate pathway genes RK11, RPE1, TAL1, and TKL1 were cloned in pB3 PGK and integrated in the locus of the respective gene, resulting in simultaneous overexpression of the genes in the xylose-fermenting S . cerevisiae strain TMB3001.

Indian J Exp Biol, 2003 Jun, 41(6), 620 - 6
Xylanase production by Ganoderma lucidum on liquid and solid state fermentation; Malarvizhi K et al.; Ganoderma lucidum, a white rot fungus, was exploited for its potentials to produce xylanase employing shake and solid-state culture conditions . Different culture conditions such as pH, temperature, carbon and nitrogen requirements for its growth and production of xylanase were optimized . The culture media pH 6.0-7.0 and temperatures 30 degrees-35 degrees C significantly promoted the growth as well as xylanase secretion into the media . Xylan and peptone were found to be the suitable carbon and nitrogen sources . Among the different agrowastes used, wheat bran was found to be the best substrate for the test fungus for the production of xylanase than sugarcane bagasse and rice bran in solid-state fermentation.

J Basic Microbiol, 2004, 44(4), 280 - 7
Biosynthesis of proteases by Rhizopus oligosporus IHS13 in low-cost medium by solid-state fermentation; Haq IU et al.; The present study describes the biosynthesis of proteases by a locally isolated mould culture of Rhizopus oligosporus IHS13 in a low-cost medium by solid-state fermentation technique . The fermentation was carried out in a low-cost medium such as sunflower meal, wheat bran and rice bran . Sunflower meal and wheat bran in a ratio of 1:1 and moistened with distilled water was found to be the best substrate for protease synthesis . All the three substrates are very cheap agricultural by-products found in Pakistan . The production of proteases in sunflower meal and rice bran was also investigated but the results were unsatisfactory . Different cultural conditions such as rate of fermentation, effect of incubation temperature, effect of pH and depth of the fermentation medium were also optimized . The maximum enzyme synthesis was found after 72 h of fermentation at a temperature of 30 degrees C . The optimum pH and depth of the medium for protease synthesis were found to be 5.0 and 20 mm respectively . The maximum enzyme biosynthesis found during the course of present studies was 7.0 U ml(-1) .

Biotechnol Lett, 2004 Jul, 26(14), 1163 - 6
Combining induced mutation and protoplasting for strain improvement of Aspergillus oryzae for kojic acid production; Wan HM et al.; By combining induced mutation, using NTG and UV irradiation, and protoplasting of a wild type strain of Aspergillus oryzae ATCC 22788, a hyper-producing strain was obtained that accumulated 41 g kojic acid l(-1) in shake-flasks, which was 100-fold higher than that in the wild type strains . Similar production of kojic acid was obtained in 5 l stirred-tank fermentations.

J Agric Food Chem, 2004 Jul 28, 52(15), 4828 - 33
Identification of fruity/fermented odorants in high-temperature-cured roasted peanuts; Didzbalis J et al.; Gas chromatography/olfactometry on a concentrate of volatiles obtained by solvent-assisted flavor evaporation (SAFE) from roasted peanuts containing a fruity/fermented off-note was used to identify the odorants responsible for the flavor defect . Freshly dug peanuts were divided into two classes, mature and immature, using pod mesocarp color, and subjected to normal (27 degrees C) and high (40 degrees C) temperature curing . Sensory evaluation of the roasted peanuts found that immature peanuts cured at high temperature contained the fruity/fermented off-note . Mature peanuts cured at high temperature and both immature and mature peanuts cured at low temperature were free of the off-note . Peanuts with the off-flavor were found to contain fruit-like esters (ethyl 2-methylpropanoate, ethyl 2-methylbutanoate, and ethyl 3-methylbutanoate) along with increased levels of short chain organic acids (butanoic, 3-methylbutanoic, and hexanoic) . These findings were confirmed by sensory evaluation of models, where the addition of these compounds produced the fruity/fermented flavor defect in a control peanut paste . This is the first time that the odorants responsible for this off-note in roasted peanuts have been identified .

Arch Anim Nutr, 2004 Jun, 58(3), 233 - 44
Agri-environmental grass hay: nutritive value and intake in comparison with hay from intensively managed grassland; Fiems LO et al.; Chemical composition, digestibility, nutritive value and intake of hay from an agri-environmental management (EH) were compared with those from hay (Lolium perenne) from an intensive management (IH) . IH was of low to moderate quality because of unfavourable weather conditions . EH was harvested mid-June of 2000 (EH1) and 2001 (EH2) on the same sward that had not received mineral fertilizer for 10 years . The EH was characterized by a species-rich botanical composition . On average, it had lower contents of protein (32%), NDF (9%) and ash (35%), and a higher concentration of water-soluble carbohydrates (117%) than IH . Digestibility of dry and organic matter, determined with sheep, was not different between IH and EH and averaged 59 and 63%, respectively . Crude fibre and NDF digestibility were lower in EH (58 and 57%, respectively) than in IH (70 and 69%, respectively) . Net energy value for lactation did not differ between IH and EH and amounted to 4.78 MJ per kg DM . True protein digested in the small intestine and rumen degraded protein balance were lower in EH (63 and -60 g per kg DM) than in IH (71 and -33 g per kg DM) . Intake of hay was investigated in Holstein-Friesian heifers and Belgian Blue double-muscled heifers (mean BW 280 +/- 22 kg and 269 +/- 21 kg, respectively), and in Belgian Blue non-lactating and non-pregnant double-muscled cows (initial BW 642 +/- 82 kg), using a cross-over design . Hay was freely available . It was supplemented with 1 kg concentrate daily . Dry matter intake from hay was higher for EH than for IH in heifers (4% and 13%, respectively in Holstein-Friesian and Belgian Blue heifers) and in cows (22%) . Hay from an agri-environmental management may be used for low-performing animals, as energy intake only exceeded maintenance requirements by 20 to 35% . Several characteristics of EH were different between years, such as dry matter digestibility, net energy value for lactation and fermentable organic matter content.

Colloids Surf B Biointerfaces, 2004 Mar 1, 34(1), 7 - 11
Effect of low frequency ultrasonic stimulation on the secretion of siboflavin produced by Ecemothecium Ashbyii; Chuanyun D et al.; The structure of the cell membrane of Ecemothecium ashbyii is considered to be one of the main obstacles in the improvement of riboflavin productivity, which impedes the release of riboflavin from the cell into the fermentation broth . The results of the present study show that stimulation on growth and riboflavin biosynthesis phase, the content of riboflavin was most obviously enhanced, as compared with the control and the total riboflavin quantity released by ultrasonic treatment . The optimal stimulation time was from 104 to 112 h and ultrasonic should be loaded every 1.5 h.

Colloids Surf B Biointerfaces, 2004 Mar 1, 34(1), 1 - 6
Research on solid-state fermentation on rice chaff with a microbial consortium; Yang YH et al.; A microbial consortium of Trichoderma reesei AS3.3711, Aspergillus niger 3.316 and Saccharomyces cerevisiaes AS2.399 was constructed to decomposed rice chaff on the basis of the characters of each microorganism and the mechanism of cellulases . In this experiment, rice chaff was pretreated before fermentation with NaOH so that the lignin structure of rice chaff was degraded and hemicellulose was dissolved partly, which remove the protection of lignin and hemicellulose on cellulose and demolish its special crystal structure . After pretreatment, rice chaff can be degraded more easily with the microbial consortium . The optimal technical paths and technological methods were achieved for intenerating rice chaff with the microbial consortium perfectly through orthogonal experiment . According to the technological methods, some experiments were done at 30 degrees C with pH 4.5 . It was found that the highest filter paper enzyme activity (FPA) was 5.64 U/g and the ratio of cellulose degradation (RCD) was 28.05%.

Colloids Surf B Biointerfaces, 2004 Jul 15, 36(2), 101 - 4
Effect of ultrasound stimulation on metabolic modulation of pyruvic acid; Chuanyun D et al.; Pyruvic acid lay on the center position of Eremothecium ashbyii metabolic fluxes, during fermentation, a high performance liquid chromatography (HPLC) method is presented for the simultaneous determination of pyruvic acid in the Eremothecium ashbyii cell culture of both with ultrasound stimulation and the control . Dry weight and riboflavin are also mentored . The results of this study show that the ultrasound stimulation can give rise to pyruvic acid change, at the same time; change of pyruvic acid concentration is correlative to that of dry weight of mycelium and content of riboflavin.

J Ind Microbiol Biotechnol, 2004 Aug, 31(7), 295 - 300 Epub 2004 Jul 15.
Flux to acetate and lactate excretions in industrial fermentations: physiological and biochemical implications; El-Mansi M; The efficiency of carbon conversion to biomass and desirable end products in industrial fermentations is diminished by the diversion of carbon to acetate and lactate excretions . In this study, the use of prototrophic and mutant strains of Escherichia coli, as well as enzyme active site directed inhibitors, revealed that flux to acetate excretion is physiologically advantageous to the organism as it facilitates a faster growth rate (mu) and permits growth to high cell densities . Moreover, the abolition of flux to acetate excretion was balanced by the excretion of lactate as well as 2-oxoglutarate, isocitrate and citrate, suggesting a 'bottle-neck' effect at the level of 2-oxoglutarate in the Krebs cycle . It is proposed that the acetate excreting enzymes, phosphotransacetylase and acetate kinase, constitute an anaplerotic loop or by-pass, the primary function of which is to replenish the Krebs cycle with reduced CoA, thus relieving the bottle-neck effect at the level of 2-oxoglutarate dehydrogenase . Furthermore, flux to lactate excretion plays a central role in regenerating proton gradient and maintaining the redox balance within the cell . The long-held view that flux to acetate and lactate excretions is merely a function of an 'over-flow' in central metabolism should, therefore, be re-evaluated.

Microbiology, 2004 Jul, 150(Pt 7), 2209 - 20
Functional analysis of the ALD gene family of Saccharomyces cerevisiae during anaerobic growth on glucose: the NADP+-dependent Ald6p and Ald5p isoforms play a major role in acetate formation; Saint-Prix F et al.; In Saccharomyces cerevisiae, acetate is formed by acetaldehyde dehydrogenase (ACDH), a key enzyme of the pyruvate dehydrogenase (PDH) bypass, which fulfils the essential task of generating acetyl-CoA in the cytosol . The role of the five members of the ACDH family (ALD genes) was investigated during anaerobic growth on glucose . Single and multiple ald Delta mutants were generated in the wine-yeast-derived V5 and laboratory CEN.PK strains and analysed under standard (YPD 5 % glucose) and wine (MS 20 % glucose) fermentation conditions . The deletion of ALD6 and ALD5 decreased acetate formation in both strains, demonstrating for the first time that the mitochondrial Ald5p isoform is involved in the biosynthesis of acetate during anaerobic growth on glucose . Acetate production of the ald4 Delta mutant was slightly decreased in the CEN.PK strain during growth on YPD only . In contrast, the deletion of ALD2 or ALD3 had no effect on acetate production . The absence of Ald6p was compensated by the mitochondrial isoforms and this involves the transcriptional activation of ALD4 . Consistent with this, growth retardation was observed in ald6 Delta ald4 Delta, and this effect was amplified by the additional deletion of ALD5 . A ald Delta null mutant, devoid of ACDH activity, was viable and produced similar levels of acetate to the ald6 Delta ald4 Delta ald5 Delta strain, excluding a role of Ald2p and Ald3p . Thus, acetate is mainly produced by the cytosolic PDH bypass via Ald6p and by a mitochondrial route involving Ald5p . An unknown alternative pathway can compensate for the loss of Ald6p, Ald4p and Ald5p.

Planta Med, 2004 Jul, 70(7), 642 - 5
Tryptanthrin content in Isatis tinctoria leaves--a comparative study of selected strains and post-harvest treatments; Oberthur C et al.; Tryptanthrin is a pharmacologically active compound in the anti-inflammatory herb Isatis tinctoria, with potent inhibitory activity on prostaglandin and leukotriene synthesis and on inducible NO synthase . The tryptanthrin content of five defined woad strains was analyzed in dependence of the time of harvest and post-harvest treatment . Tryptanthrin was determined by a validated ESI-LC-MS isotope dilution assay with d(8)-tryptanthrin as internal standard . The tryptanthrin concentration in freeze-dried leaf samples was low . Drying at ambient temperature led to a significant increase of tryptanthrin concentration, but the highest concentrations were found when leaves were dried at 40 degrees C . Tryptanthrin content in fermented woad leaves was below the limit of quantification . Tryptanthrin appears thus to be a product of post-harvest processes, but details of its formation remain to be elucidated.

Altern Med Rev, 2004 Jun, 9(2), 208 - 210
Monascus purpureus (red yeast rice) - Monograph; An ethanologenic yeast exhibiting unusual metabolism in the fermentation of lignocellulosic hexose sugars; Forest Products Biotechnology, Department of Wood Science, University of British Columbia, 2424 Main Mall, Vancouver, BC, Canada, V6T 1Z4Three lignocellulosic substrate mixtures {liquid fraction of acid-catalyzed steam-exploded softwood, softwood spent sulfite liquor (SSL) and hardwood SSL} were separately fermented by the industrially employed SSL-adapted strain Tembec T1 and a natural galactose-assimilating isolate (Y-1528) of Saccharomyces cerevisiae to compare fermentative efficacy . Both strains were confirmed as S . cerevisiae via molecular genotyping . The performance of strain Y-1528 exceeded that of Tembec T1 on all three substrate mixtures, with complete hexose sugar consumption ranging from 10 to 18 h for Y-1528, vs 24 to 28 h for T1 . Furthermore, Y-1528 consumed galactose prior to glucose and mannose, in contrast to Tembec T1, which exhibited catabolite repression of galactose metabolism . Ethanol yields were comparable regardless of the substrate utilized . Strains T1 and Y-1528 were also combined in mixed culture to determine the effects of integrating their distinct metabolic capabilities during defined hexose sugar and SSL fermentations . Sugar consumption in the defined mixture was accelerated, with complete exhaustion of hexose sugars occurring in just over 6 h . Galactose was consumed first, followed by glucose and mannose . Ethanol yields were slightly reduced relative to pure cultures of Y-1528, but normal growth kinetics was not impeded . Sugar consumption in the SSLs was also accelerated, with complete utilization of softwood- and hardwood-derived hexose sugars occurring in 6 and 8 h, respectively . Catabolite repression was absent in both SSL fermentations.

J Ind Microbiol Biotechnol, 2004 Aug, 31(7), 311 - 22 Epub 2004 Jul 13.
The application of multi-parameter flow cytometry to the study of recombinant Escherichia coli batch fermentation processes; Lewis G et al.; Multi-parameter flow cytometric techniques coupled with dual colour fluorescent staining were used to study the physical and metabolic consequences of inclusion body formation in batch cultures of the recombinant Escherichia coli strain MSD3735 . This strain contains a plasmid coding for the isopropylthiogalactopyranoside-inducible model eukaryotic protein AP50 . It is known that the synthesis of foreign proteins at high concentrations can exert a severe metabolic stress on the host cell and that morphological changes can occur . In this work, using various points of induction, it was shown that inclusion body formation is followed immediately by measurable changes in the characteristic intrinsic light scatter patterns for the individual cell (forward scatter, 90 degrees side scatter) and a concomitant progressive change in the individual cell physiological state with respect to both cytoplasmic membrane polarisation and permeability . This work establishes flow cytometry as a potentially valuable tool for monitoring recombinant fermentation processes, providing important information for scale-up . Further, we discuss the possibility of optimising inclusion body formation by manipulating the fermentation conditions based on these rapid "real-time" measurements.

Indian J Exp Biol, 2003 Aug, 41(8), 870 - 4
Influence of nutritional supplements on keratinolysis by Amycolatopsis keratinophila; Al-Musallam AA et al.; Keratinolytic potential of A . keratinophila (DSM 44409T), a newly described Amycolatopsis sp . isolated from cultivated soil in Kuwait, was demonstrated using keratinazure as the sole source of carbon and nitrogen as estimated by gel diffusion assay . Effects of 12 various nutritional supplements on the keratinolytic and azocollytic activities were determined . NH4H2PO4 and KNO3 in the medium supported a significantly higher keratinolytic activity than other supplements . However, azocollytic activities in all the supplemented media and the control were same . Best combination of carbon and nitrogen supplements (galactose and NH4H2PO4 respectively) used to evaluate the dynamics of growth and enzymes (keratinase and protease) activities of the isolate revealed a luxuriant growth with optimal keratinolytic activity occurring during the log phase . Other parameters of the fermentation medium, including pH, biomass accumulation, total protein and free amino acid concentrations were also studied.

Plant Biol (Stuttg), 2004 Jul, 6(4), 375 - 86
Seed development and differentiation: a role for metabolic regulation; Borisjuk L et al.; During seed growth, the filial organs, Vicia embryos and barley endosperm, differentiate into highly specialized storage tissues . Differentiation is evident on structural and morphological levels and is reflected by the spatial distribution of metabolites . In Vicia embryos, glucose is spatially correlated to mitotic activity whereas elongating and starch accumulating cells contain high levels of sucrose . Seed development is also regulated by phytohormones . In pea seeds, GA-deficiency stops seed growth before maturation . In Arabidopsis seeds, ABA regulates differentiation and inhibits cell division activity . The ABA pathway, in turn, is linked to sugar responses . In young Vicia embryos, invertases in maternal tissues control both concentration and composition of sugars . Embryonic and endospermal transfer cell formation represents an early differentiation step . Establishing an epidermis-localised sucrose uptake system renders the embryo independent from maternal control . cDNA array analysis in barley seeds revealed a massive transcriptional re-programming of gene expression during the transition stage, when gene clusters related to transport and energy metabolism are highly transcribed . Sucrose represents a signal for differentiation and up-regulates storage-associated gene expression . Sucrose signalling involves protein phosphorylation . Sucrose non-fermenting-1-related protein kinases are apparently induced in response to high cellular sucrose, and could act as mediators of sucrose-specific signals . Energy metabolism changes during seed development . In Vicia embryos metabolic responses upon hypoxia and low energy charge levels are characteristic for young undifferentiated stages when energy demand and respiration are high . During the transition stage, the embryo becomes adapted to low energy availability and metabolism becomes energetically more economic and tightly controlled . These adaptations are embedded in the embryo's differentiation program and coupled with photoheterotrophic metabolism . In Vicia cotyledons, ATP content increases in a development-dependent pattern and is associated with the greening process . The main role of seed photosynthesis is to increase internal O2 contents and to control biosynthetic fluxes by improving energy supply . Copyright Georg Thieme Verlag KG Stuttgart

Appl Microbiol Biotechnol, 2004 Dec, 66(2), 153 - 9 Epub 2004 Dec.
Biotechnological lycopene production by mated fermentation of Blakeslea trispora; Lopez-Nieto MJ et al.; A semi-industrial process (800-l fermentor) for lycopene production by mated fermentation of Blakeslea trispora plus (+) and minus (-) strains has been developed . The culture medium was designed at the flask scale, using a program based on a genetic algorithm; and a fermentation process by means of this medium was developed . Fermentation involves separate vegetative phases for (+) and (-) strains and inoculation of the production medium with a mix of both together . Feeding with imidazole or pyridine, molecules known to inhibit lycopene cyclase enzymatic activity, enhanced lycopene accumulation . Different raw materials and physical parameters, including dissolved oxygen, stirring speed, air flow rate, temperature, and pH, were checked in the fermentor to get maximum lycopene production . Typical data for the fermentation process are presented and discussed . This technology can be easily scaled-up to an industrial application for the production of this carotenoid nowadays widely in demand.

Ann N Y Acad Sci, 2004 Jun, 1019, 195 - 9
The aging/precancerous gastric mucosa: a pilot nutraceutical trial; Marotta F et al.; The aim of this study was to test the effect of antioxidant supplementation on enzymatic abnormalities and free radical-modified DNA adducts associated with premalignant changes in the gastric mucosa of elderly patients with HP-negative atrophic gastritis (CAG) . Sixty patients with atrophic gastritis and intestinal metaplasia underwent a nutritional interview and a gastroscopy with multiple biopsy samples in the antrum that were processed for histology and for assaying: alpha-tocopherol, MDA, xanthine oxidase (XO), ornithine decarboxylase (ODC), and 8-OHdG . Patients were randomly allocated into three matched groups and supplemented for 6 months with (1) vitamin E, 300 mg/day; (2) multivitamin, two tablets t.i.d.; and (3) Immun-Age 6 g/day nocte (ORI, Gifu, Japan), a certified fermented papaya preparation with basic science-validated antioxidant/immunomodulant properties . Ten dyspeptic patients served as controls . Histology and biochemistry were blindly repeated at 3 and 6 months . CAG patients showed a significantly (P <.05) increased level of mucosal MDA and XO concentration that were reverted to normal by each supplementation (P <.05) . All supplements caused a significant decrease of ODC (P <.01), but Immun-Age yielded the most effective (P < 0.05) and was the only one significantly decreasing 8-OhdG (P < 0.05) . These data suggest that antioxidant supplementation, and, namely, Immun-Age, might be potential chemopreventive agents in HP-eradicated CAG patients and especially in the elderly population.

J Biotechnol, 2004 Aug 5, 111(3), 297 - 309
Anaerobic bio-hydrogen production from ethanol fermentation: the role of pH; Hwang MH et al.; Hydrogen was produced by an ethanol-acetate fermentation at pH of 5.0 +/- 0.2 and HRT of 3 days . The yield of hydrogen was 100-200 ml g Glu(-1) with a hydrogen content of 25-40% . This fluctuation in the hydrogen yield was attributed to the formation of propionate and the activity of hydrogen utilizing methanogens . The change in the operational pH for the inhibition of this methanogenic activity induced a change in the main fermentation pathway . In this study, the main products were butyrate, ethanol and propionate, in the pH ranges 4.0-4.5, 4.5-5.0 and 5.0-6.0, respectively . However, the activity of all the microorganisms was inhibited below pH 4.0 . Therefore, pH 4.0 was regarded as the operational limit for the anaerobic bio-hydrogen production process . These results indicate that the pH plays an important role in determining the type of anaerobic fermentation pathway in anaerobic bio-hydrogen processes.

Bioresour Technol, 2004 Nov, 95(2), 121 - 8
Aerobic composting of chips from clear-cut trees with various co-materials; Suzuki T et al.; Swollen chips made from trees felled during clear-cutting were composted with various organic and inorganic materials in an aerobic composting reactor for 5 months and then piled for 5 months . The organic materials included chicken feces, urea, nitrogenous lime (calcium cyanamide, manure), and material rapidly composted from food garbage in 24-h bacterial fermentation, while the inorganic materials were coal ash and volcanic ash . In this paper, we first attempt to estimate the quality and degree of maturity of each compost from its chemical properties . Furthermore, we try to calculate the maturity of the fermented wood chip composts from their mixture ratio of the initial materials by multiple linear regression analysis . We measured changes in the C/N ratio, nitrate nitrogen (NO3-N) content, percentage of humic acid in the alkali soluble fraction (PQ), cation exchange capacity, pH, and EC during the composting period . The degrees of maturity of the composts were estimated via a plant growth test using Chinese cabbage . We found that the CN ratio, NO3-N concentration, and PQ were suitable for estimating the degree of maturity of wood chip composts . For maturity, the CN ratio should be less than 14, the PQ more than 66.2, and the NO3-N concentration more than 853 mgkg-1 . We devised an equation to estimate the degree of maturity after 10 months by a multiple linear regression analysis from the mixing ratio of wood chips and the co-composted materials . From the multiple linear regression analysis, the above three indices of compost maturity could be estimated from the mixing ratio of the initial materials . This equation should enable us to determine the degree of compost maturity after 10 months based on the initial mixing ratio.

Lett Appl Microbiol, 2004, 39(2), 199 - 206
Manipulating the pyruvate dehydrogenase bypass of a multi-vitamin auxotrophic yeast Torulopsis glabrata enhanced pyruvate production; Liu LM et al.; AIMS: To investigate the relationship between the activity of pyruvate dehydrogenase (PDH) bypass and the production of pyruvate of a multi-vitamin auxotrophic yeast Torulopsis glabrata . METHODS AND RESULTS: Torulopsis glabrata CCTCC M202019, a multi-vitamin auxotrophic yeast that requires acetate for complete growth on glucose minimum medium, was selected after nitrosoguanidine mutagenesis of the parent strain T . glabrata WSH-IP303 screened in previous study {Li et al . (2001) Appl . Microbiol . Biotechnol . 55, 680-685} . Strain CCTCC M202019 produced 21% higher pyruvate than the parent strain and was genetically stable in flask cultures . The activities of the pyruvate metabolism-related enzymes in parent and mutant strains were measured . Compared with the parent strain, the activity of pyruvate decarboxylase (PDC) of the mutant strain CCTCC M202019 decreased by roughly 40%, while the activity of acetyl-CoA synthetase (ACS) of the mutant increased by 103.5 or 57.4%, respectively, in the presence or absence of acetate . Pyruvate production by the mutant strain CCTCC M202019 reached 68.7 g l(-1) at 62 h (yield on glucose of 0.651 g g(-1)) in a 7-l jar fermentor . CONCLUSIONS: The increased pyruvate yield in T . glabrata CCTCC M202019 was due to a balanced manipulation of the PDH bypass, where the shortage of cytoplasmic acetyl-CoA caused by the decreased activity of PDC was properly compensated by the increased activity of ACS . SIGNIFICANCE AND IMPACT OF THE STUDY: Manipulating the PDH bypass may provide an alternative approach to enhance the production of glycolysis-related metabolites.

J Nutr Sci Vitaminol (Tokyo), 2004 Apr, 50(2), 114 - 20
Promotion of bone formation by fermented soybean (Natto) intake in premenopausal women; Katsuyama H et al.; A therapeutic agent of vitamin K2 is approved for the treatment of osteoporosis in Japan . However, little is known about the efficacy of dietary intake of vitamin K2 for bone health . We compared the effects of various levels of fermented soybeans (Natto) intake, which contains plenty of vitamin K2, on bone stiffness and bone turnover markers in healthy premenopausal women . Seventy-three healthy premenopausal women were randomly divided into four groups matched for age and parity categories . Natto was supplied as follows: Group 1 (no intake), Group 2 (once per month), Group 3 (once per week) and Group 4 (three times per week) . Subjects took Natto at a lunch for 1 y, and the stiffness index by quantitative ultrasound and bone turnover markers were assessed at baseline, 6 mo and 1 y . There was no statistical difference in the stiffness index during the 1 y observation . However, bone specific alkaline phosphatase (BAP) in Group 4 was higher than that in Group 3 at 1 y and undercarboxylated osteocalcin (Glu) in Group 4 was significantly lower than those in Groups 1, 2 and 3 at 6 mo . Logistic regression analysis showed that the risk of reduction of bone formation markers declined to 0.07 in Group 4 based on that in Group 1 . In premenopausal women who had to keep the stiffness index as high as possible before menopause, Natto intake may have contributed to the promotion of bone formation.

Ann N Y Acad Sci, 2004 Jun, 1018, 173 - 82
Alcohol alters rat adrenomedullary function and stress response; Patterson-Buckendahl P et al.; Most alcohol researchers do not address the effects of intoxication on the sympatho-adrenomedullary system response to stressful situations . We previously determined that rats consuming nearly 9 g ethanol (EtOH) per kg body weight per day in liquid diet form for 1 week increased adrenal gene expression of enzymes for catecholamine synthesis that was further elevated by acute IMMO . We hypothesized that the response to chronic mild stressors would also be altered after consumption of lower concentrations of EtOH in drinking water . Two experiments were conducted: 10% w/v for 4 weeks or 6% w/v for 7 weeks +/- wire mesh restraint (WMR) . These were compared with ad libitum (adlib) and pair-fed control rats . Adrenal gene expression of catecholamine synthesizing enzymes was assayed . Tyrosine hydroxylase gene expression was elevated 80% to 90% by alcohol consumption in both experiments (P < 0.001) compared with adlib control rats . Dopamine betab-hydroxylase and phenylethanolamine-N-methyl transferase gene expressions were unaffected by 10% alcohol (P > 0.05) but were increased by 6% alcohol (P < 0.01) . WMR decreased already elevated gene expression of all three enzymes . Pair feeding to 6% EtOH drinkers also increased gene expression for the three enzymes but was decreased by WMR, although not to levels of adlib rats . Increased gene expression for adrenal synthesis of catecholamines in response to repeated alcohol consumption increases the likelihood that the subject can respond physiologically to acute or chronic stress . This may have life-saving consequences in humans and in animals known to consume fermented materials and may contribute to increased aggressive behavior.

Int J Food Microbiol, 2004 Aug 15, 95(1), 67 - 78
Effect of weak acid preservatives on growth of bakery product spoilage fungi at different water activities and pH values; Suhr KI et al.; Inhibition of spoilage organisms from bakery products by weak acid preservatives in concentrations of 0%, 0.003%, 0.03% and 0.3% (w/v) was investigated experimentally on a substrate media with water activity (a(w)) and pH ranging from sourdough-fermented acidic rye bread to alkaline intermediate moisture sponge cake types (a(w) 0.80-0.95, pH 4.7-7.4) . Initially, rye bread conditions (a(w) 0.94-0.97 and pH 4.4-4.8) in combination with calcium propionate were investigated . Results showed that the highest concentration of propionate (0.3%) at all conditions apart from high a(w) (0.97) and high pH (4.8) totally inhibited fungal growth for a 2-week period, with the exception of Penicillium roqueforti, Penicillium commune and Eurotium rubrum . Characteristically for the major spoiler of rye bread, P . roqueforti, all three isolates tested were stimulated by propionate and the stimulation was significantly enhanced at high water activity levels . The effect of propionate on production of secondary metabolites (mycophenolic acid, rugulovasine, echinulin, flavoglaucin) was also studied, and variable or isolate dependent results were found . Subsequently, a screening experiment representing a wider range of bakery products was conducted using calcium propionate, potassium sorbate and sodium benzoate . The obtained data was modelled using survival analysis to determine 'spoilage-free time' for the fungi . At the low a(w) level (0.80) only Eurotium species grew within the test period of 30 days . Higher water activity levels as well as higher pH values decreased spoilage-free times of the fungi . The preservative calcium propionate was less effective than potassium sorbate and sodium benzoate.

Food Chem Toxicol, 2004 Sep, 42(9), 1531 - 42
A review of the clinical toleration studies of polydextrose in food; Flood MT et al.; Polydextrose is a non-digestible 1 kcal/g polysaccharide used primarily as a sugar replacer and dietary fiber in foods . At typical use levels, polydextrose provides physiological effects similar to those of other dietary fibers . However, excessive consumption of non-digestible carbohydrates can lead to gastrointestinal distress . Nine clinical studies were conducted with polydextrose to evaluate the extent of such symptoms . These studies determined laxation endpoints in adults and children, and showed that polydextrose was better tolerated than most other low digestible carbohydrates (e.g . polyols) . This is because of a higher molecular weight and partial colonic fermentation, leading to a lower risk of osmotic diarrhea . After evaluating these studies, the Joint FAO/WHO Expert Committee on Food Additives (JECFA) and the European Commission Scientific Committee for Food (EC/SCF) concluded that polydextrose has a mean laxative threshold of approximately 90 g/d (1.3 g/kg bw) or 50 g as a single dose.

J Ren Nutr, 2004 Jul, 14(3), 149 - 56
Calcium-enriched bread for treatment of uremic hyperphosphatemia; Babarykin D et al.; OBJECTIVE: To assess phosphate-binding efficacy of a new food product, bread with unusually high calcium content (Ca-bread) . DESIGN AND SETTING: A randomized parallel group trial in the university hospital outpatient dialysis unit . PATIENTS: Fifty-three randomly selected uremic patients who met the following inclusion criteria: (1) required maintenance hemodialysis treatment, (2) were not to receive vitamin D throughout the study, (3) were nondiabetic, and (4) were diagnosed with hyperphosphatemia . INTERVENTION: Fifty-three patients were randomized into 2 groups: control group (n = 26), which received calcium acetate as a phosphate binder throughout the study, and Ca-bread group (n = 27), which, after a 2-week washout period, received Ca-bread containing 2.5% of elemental calcium (by weight), which served as a phosphate binder . Bread was made using wheat flour, calcium carbonate, and fermented buttermilk . The amount of elemental calcium used as a phosphate binder was similar in both groups . Observation of both groups lasted 14 weeks . RESULTS: Mean serum phosphate concentration at randomization was 2.11 +/- 0.14 mmol/L in the control group and 2.20 +/- 0.13 mmol/L in the Ca-bread group . Mean serum calcium concentration at randomization was 2.12 +/- 0.21 mmol/L in the control group and 2.14 +/- 0.11 mmol/L in the Ca-bread group . The Ca-bread group patients' predialysis phosphate concentration decreased to a mean of 1.67 +/- 0.18 mmol/L (P <.05), and their mean calcium concentration increased to 2.27 +/- 0.11 mmol/L (P = NS) . In the control group, neither value changed significantly from the original readings . After the hemodialysis session, the mean serum calcium concentration in the control group and the Ca-bread group increased by 7.5% and 7.9%, respectively (P = NS) . Mean phosphate concentration simultaneously decreased to nearly 1/2 its original predialysis value in both groups . Ca-bread group patients saw a decrease in the mean phosphate concentration (from predialysis to postdialysis values) that was 13.8% greater than that of the control group (P = NS) . CONCLUSION: A new form of calcium-containing phosphate binder was developed: Ca-bread with an elemental calcium content of 2.5% . Ca-bread allows for effective amelioration of hyperphosphatemia without inducing hypercalcemia . Furthermore, patient compliance may increase if hyperphosphatemia can be treated by consuming bread with an elevated calcium content.

Nutr Cancer, 2004, 48(2), 189 - 97
Induction of apoptosis by Antrodia camphorata in human premyelocytic leukemia HL-60 cells; Hseu YC et al.; Antrodia camphorata (A . camphorata) is well known in Taiwan as a traditional Chinese medicine, and it has been shown to exhibit antioxidant effects . In this study, the ability of A . camphorata to induce apoptosis was studied in cultured human premyelocytic leukemia HL-60 cells . Treatment of the HL-60 cells with a variety of concentrations of the fermented culture broth of A . camphorata (25-150 microg/ml) resulted in dose- and time-dependent sequences of events marked by apoptosis, as shown by loss of cell viability, chromatin condensation, and internucleosomal DNA fragmentation . Furthermore, apoptosis in the HL-60 cells was accompanied by the release of cytochrome c, activation of caspase-3, and specific proteolytic cleavage of poly (ADP-ribose) polymerase (PARP) . This increase in A . camphorata-induced apoptosis was also associated with a reduction in the levels of Bcl-2, a potent cell-death inhibitor, and an increase in those of the Bax protein, which heterodimerizes with and thereby inhibits Bcl-2 . The data suggest that A . camphorata exerts antiproliferative action and growth inhibition on HL-60 cells through apoptosis induction and that it may have anticancer properties valuable for application in drug products.

Br J Nutr, 2004 Jul, 92(1), 95 - 104
Inter-relationship of microbial activity, digestion and gut health in the rabbit: effect of substituting fibre by starch in diets having a high proportion of rapidly fermentable polysaccharides; Gidenne T et al.; Caecal microbial activity, digestion and gut health were analysed in the young rabbit, in response to fibre substitution by starch, in diets with high proportions of rapidly fermentable polysaccharides (pectins+hemicelluloses:acid-detergent fibre (ADF) ratio of 1.7) . A range of five diets corresponding to a 60 % linear reduction of the ADF level (230 to 92 g ADF/kg) without changes in the fibre quality, and to a corresponding linear increase in dietary starch, was given ad libitum to young rabbits from 18 d until 70 d of age . A one half reduction of the ADF level resulted in a sharp increase in energy digestibility (+25 units), associated with a lower feed intake (-35 %) and to an increase of the mean retention time (+6 h) in the whole digestive tract . Despite large variations in the fibre intake (20 to 59 g ADF/d), the fibre digestive efficiency remained similar among the five diets . Starch ileal concentrations were low after 4 weeks of age (<5 %), and variations with age were significant when the dietary starch level was over 19 % . A 65 % lower biomass production was measured when the ADF level progressed from 230 to 165 g/kg, and no precise relationship was found with fermentative activity . Reducing the fibre intake led to a linear decrease of caecal volatile fatty acids concentrations, and to higher pH and NH3 levels . An increased occurrence of mortality by diarrhoea was registered with the lowest fibre intake . It can be concluded that a sufficient supply of fibre, with high proportions in rapidly fermentable polysaccharides, stimulates the maturation of microbial activity and reduces the occurrence of diarrhoea.

Ter Arkh, 2004, 76(5), 32 - 5
{In Process Citation}; Microbial stimulation by Mycoplasma fermentans synergistically amplifies IL-6 release by human lung fibroblasts in response to residual oil fly ash (ROFA) and nickel; Department of Environmental and Occupational Health, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USAMycoplasma (MP), such as the species M . fermentans, possess remarkable immunoregulatory properties and can potentially establish chronic latent infections with little signs of disease . Atmospheric particulate matter (PM) is a complex and diverse component of air pollution associated with adverse health effects . We hypothesized that MP modulate the cellular responses induced by chemical stresses such as residual oil fly ash (ROFA), a type of PM rich in transition metals . We assessed the release of interleukin-6 (IL-6), a prototypic immune-modulating cytokine, in response to PM from different sources in human lung fibroblasts (HLF) deliberately infected with M . fermentans . We found that M . fermentans and ROFA together synergistically stimulated production of IL-6 compared to either stimuli alone . Compared to several other PM, ROFA appeared most able to potentiate IL-6 release . The potentiating effect of live MP infection could be mimicked by M . fermentans-derived macrophage-activating lipopeptide-2 (MALP-2), a known Toll-like receptor-2 agonist . The aqueous fraction of ROFA also contained potent IL-6 inducing activity in concert with MALP-2, and exposure to several defined metal salts indicated that Ni and, to a lesser extent V, (but not Cu) could synergistically act with MALP-2 to induce IL-6 . These data indicate that microorganisms like MP can interact with environmental stimuli such as PM-derived metals to synergistically activate signaling pathways that control lung cell cytokine production and, thus, can potentially modulate adverse health effects of PM exposure.

Asia Pac J Clin Nutr, 2004, 13(2), 156 - 61
Contemporary food technology and its impact on cuisine; Lupien JR et al.; This article contains a general review of current food technologies and their effect on the quality and safety of foods . The uses of these technologies in the context of current and projected world population, urbanization prevention of food losses, preservation and trade of foods, domestically and internationally are briefly reviewed . Technologies discussed are related to food storage, refrigeration, frozen foods, milling and baking, canning, pickling, extrusion processes, fermentation, and to foods and food ingredients such as fruits and vegetables, animal products, fat and oils, canned products, and food additives.

Asia Pac J Clin Nutr, 2004, 13(2), 147 - 55
Traditional Chinese food technology and cuisine; Li JR et al.; From ancient wisdom to modern science and technology, Chinese cuisine has been established from a long history of the country and gained a global reputation of its sophistication . Traditional Chinese foods and cuisine that exhibit Chinese culture, art and reality play an essential role in Chinese people's everyday lives . Recently, traditional Chinese foods have drawn a great degree of attention from food scientists and technologists, the food industry, and health promotion institutions worldwide due to the extensive values they offer beyond being merely another ethnic food . These traditional foods comprise a wide variety of products, such as pickled vegetables, salted fish and jellyfish, tofu and tofu derived products, rice and rice snack foods, fermented sauces, fish balls and thousand-year-old eggs . An overview of selected popular traditional Chinese foods and their processing techniques are included in this paper . Further development of the traditional techniques for formulation and production of these foods is expected to produce economic, social and health benefits.

Biochem J, 2004 Oct 1, 383(Pt 1), 171 - 8
Gts1p stabilizes oscillations in energy metabolism by activating the transcription of TPS1 encoding trehalose-6-phosphate synthase 1 in the yeast Saccharomyces cerevisiae; Xu Z et al.; We reported previously that Gts1p regulates oscillations of heat resistance in concert with those of energy metabolism in continuous cultures of the yeast Saccharomyces cerevisiae by inducing fluctuations in the levels of trehalose, but not in those of Hsp104 (heat shock protein 104) . Further, the expression of TPS1, encoding trehalose-6-phosphate synthase 1, and HSP104 was activated by Gts1p in combination with Snf1 kinase, a transcriptional activator of glucose-repressible genes, in batch cultures under derepressed conditions . Here we show that, in continuous cultures, the mRNA level of TPS1 increased 6-fold in the early respiro-fermentative phase, while that of HSP104 did not change . The expression of SUC2, a representative glucose-repressible gene encoding invertase, also fluctuated, suggesting the involvement of the Snf1 kinase in the periodic activation of these genes . However, this possibility was proven to be unlikely, since the oscillations in both TPS1 and SUC2 mRNA expression were reduced by approx . 3-fold during the transient oscillation in gts1Delta (GTS1-deleted) cells, in which the energy state determined by extracellular glucose and intracellular adenine nucleotide levels was comparable with that in wild-type cells . Furthermore, neither the mRNA level nor the phosphorylation status of Snf1p changed significantly during the oscillation . Thus we suggest that Gts1p plays a major role in the oscillatory expression of TPS1 and SUC2 in continuous cultures of Saccharomyces cerevisiae, and hypothesized that Gts1p stabilizes oscillations in energy metabolism by activating trehalose synthesis to facilitate glycolysis at the shift from the respiratory to the respiro-fermentative phase.

Gene, 2004 Jul 7, 336(1), 81 - 91
Identification and characterization of traE genes of Spiroplasma kunkelii; Bai X et al.; Four traE homologs, designated traE1, traE2, traE3 and traE4, were identified and amplified from the genome of the leafhopper-transmitted corn stunt pathogen Spiroplasma kunkelii and were predicted to encode membrane-bound adenine tri-phosphatases (ATPases) . Deduced proteins of all traE genes have 62.3% to 89.9% similarity to the conserved VirB4 domain that is frequently a component of type IV secretory pathways involved in intracellular trafficking and secretion of DNA and proteins . In phylogenetic analysis, TraE homologs of S . kunkelii, Mycoplasma pulmonis and Mycoplasma fermentans cluster together and are more similar to TraE proteins of Gram-positive bacteria than to those of Gram-negative bacteria, thereby resembling the 16S rRNA phylogeny . Gene traE2 was most conserved whereas the presence of the three other traE genes varied among S . kunkelii strains, M2, CS-2B, FL-80 and PU8-17 . Further, traE1 and traE2 appeared to be located on the chromosome, and traE3 and traE4 genes on plasmids of S . kunkelii strain M2 . Transcripts of the spiralin gene and traE2 genes were detected on Northern blots containing total ribonucleic acids (RNA) of S . kunkelii cultures and S . kunkelii-infected plants and insects, in which traE2 appeared to be of a larger transcription unit . Full-length expression products of the other traE genes were not detected . S . kunkelii traE genes could be involved in S . kunkelii cell morphogenesis, adhesion and DNA recombination.

Crit Rev Biotechnol, 2003, 23(4), 267 - 302
Oxidative stress in submerged cultures of fungi; Bai Z et al.; It has been known for many years that oxygen (O2) may have toxic effects on aerobically growing microorganisms, mainly due to the threat arising from reactive oxygen species (ROS) . In submerged culture industrial fermentation processes, maintenance of adequate levels of O2 (usually measured as dissolved oxygen tension (DOT)) can often be critical to the success of the manufacturing process . In viscous cultures of filamentous cultures, actively respiring, supplying adequate levels of O2 to the cultures by conventional air sparging is difficult and various strategies have been adopted to improve or enhance O2 transfer . However, adoption of those strategies to maintain adequate levels of DOT, that is, to avoid O2 limitation, may expose the fungi to potential oxidative damage caused by enhanced flux through the respiratory system . In the past, there have been numerous studies investigating the effects of DOT on fungal bioprocesses . Generally, in these studies moderately enhanced levels of O2 supply resulted in improvement in growth, product formation and acceptable morphological changes, while the negative impact of higher levels of DOT on morphology and product synthesis were generally assumed to be a consequence of "oxidative stress." However, very little research has actually been focused on investigation of this implicit link, and the mechanisms by which such effects might be mediated within industrial fungal processes . To elucidate this neglected topic, this review first surveys the basic knowledge of the chemistry of ROS, defensive systems in fungi and the effects of DOT on fungal growth, metabolism and morphology . The physiological responses of fungal cells to oxidative stress imposed by artificial and endogenous stressors are then critically reviewed . It is clear that fungi have a range of methods available to minimize the negative impacts of elevated ROS, but also that development of the various defensive systems or responses, can itself have profound consequences upon many process-related parameters . It is also clear that many of the practically convenient and widely used experimental methods of simulating oxidative stress, for example, addition of exogenous menadione or hydrogen peroxide, have effects on fungal cultures quite distinct from the effects of elevated levels of O2, and care must thus be exercised in the interpretation of results from such studies . The review critically evaluates our current understanding of the responses of fungal cultures to elevated O2 levels, and highlights key areas requiring further research to remedy gaps in knowledge.

Appl Microbiol Biotechnol, 2004 Jul, 65(1), 74 - 83 Epub 2004 May 20.
Transcriptional analysis of genes for energy catabolism and hydrolytic enzymes in the filamentous fungus Aspergillus oryzae using cDNA microarrays and expressed sequence tags; Maeda H et al.; Aspergillus oryzae is a fungus used extensively in the fermentation industry . We constructed cDNA microarrays comprising 2,070 highly expressed cDNAs selected from the approximately 6,000 non-redundant expressed sequence tags (ESTs) in the A . oryzae EST database . Using the cDNA microarrays, we analyzed the gene expression profiles of A . oryzae cells grown under the glucose-rich (AC) and glucose-depleted (AN) liquid culture conditions used during the construction of the EST database . The sets of genes identified by the cDNA microarray as highly expressed under each culture condition agreed well with the highly redundant ESTs obtained under the same conditions . In particular, transcription levels of most catabolic genes of the glycolytic pathway (EMP) and tricarboxylic acid (TCA) cycle were higher under AC than AN conditions, suggesting that A . oryzae uses both EMP and TCA for glucose metabolism under AC conditions . We further studied the expression of genes encoding hydrolytic enzymes and enzymes involved in energy catabolism by using three industrial solid-phase biomass media, including wheat-bran . The wheat-bran culture gave the richest gene expression profile of hydrolytic enzymes and the lowest expression levels of catabolic genes (EMP, TCA) among the three media tested . The low expression levels of catabolic genes in the wheat-bran culture may release catabolite repression, consequently leading to the rich expression profiles of the hydrolytic enzymes.

Appl Microbiol Biotechnol, 2004 Oct, 65(5), 530 - 7 Epub 2004 Jun 25.
Production of fructosyl transferase by Aspergillus oryzae CFR 202 in solid-state fermentation using agricultural by-products; Sangeetha PT et al.; Fructosyl transferase (FTase) production by Aspergillus oryzae CFR 202 was carried out by solid-state fermentation (SSF), using various agricultural by-products like cereal bran, corn products, sugarcane bagasse,cassava bagasse (tippi) and by-products of coffee and tea processing . The FTase produced was used for the production of fructo-oligosaccharides (FOS), using 60% sucrose as substrate . Among the cereal bran used, rice bran and wheat bran were good substrates for FTase production by A . oryzae CFR 202 . Among the various corn products used, corn germ supported maximum FTase production, whereas among the by-products of coffee and tea processing used, spent coffee and spent tea were good substrates, with supplementation of yeast extract and complete synthetic media . FTase had maximum activity at 60 degrees C and pH 6.0 . FTase was stable up to 40 degrees C and in the pH range 5.0-7.0 . Maximum FOS production was obtained with FTase after 8 h of reaction with 60% sucrose . FTase produced by SSF using wheat bran was purified 107-fold by ammonium sulphate precipitation (30-80%), DEAE cellulose chromatography and Sephadex G-200 chromatography . The molecular mass of the purified FTase was 116.3 kDa by SDS-PAGE . This study indicates the potential for the use of agricultural by-products for the efficient production of FTase enzyme by A . oryzae CFR 202 in SSF, thereby resulting in value addition of those by-products.

Eur J Clin Nutr, 2004 Jul, 58(7), 1046 - 55
Short-term digestive tolerance of different doses of NUTRIOSE FB, a food dextrin, in adult men; van den Heuvel EG et al.; OBJECTIVE: To determine the tolerance of increasing dosages of an incompletely hydrolysed and/or incompletely absorbed food dextrin coming from wheat starch, NUTRIOSE FB, at daily levels of 10 and 15 g up to 60 and 80 g, respectively . DESIGN: A randomized, double-blind, multiple dose, placebo-controlled, combined crossover and parallel trial . SETTING: The metabolic ward of TNO Nutrition and Food Research . SUBJECTS: A total of 20 healthy men (age 31.7 +/- 9.1 y; BMI 24.5 +/- 2.9 kg/m2) . INTERVENTION: One group of 10 subjects consumed on top of their diet 10, 30 and 60 g of NUTRIOSE FB or maltodextrin (placebo) daily . The other group of 10 subjects consumed 15, 45 and 80 g daily . Each dose was consumed for 7 days . RESULTS: Compared with placebo, flatulence occurred more frequently over the last 6 days on 30, 60 or 80 g/day of NUTRIOSE FB (P < 0.05) . During the last 24 h, that is, days 6-7, of 60 and 80 g/day of NUTRIOSE FB, the frequency of flatulence was even higher (P < 0.05) . During the last 24 h on a daily dose of 60 g NUTRIOSE FB, the frequency of defecation decreased (P < 0.05) . Bloating occurred more often during the last 24 h on 80 g/day of NUTRIOSE FB (P < 0.05) . None of the doses of NUTRIOSE FB resulted in diarrhoea . Compared to baseline levels, breath H2 excretion, which was only measured after a week with 10 and 15 g of NUTRIOSE FB daily, increased (P < 0.05) . However, no difference in area under the curve was found . CONCLUSIONS: NUTRIOSE FB is a fermentable carbohydrate and is well tolerated up to a dose of 45 g daily . Higher daily dosages (60 and 80 g) may result in flatulence, but does not result in diarrhoea . SPONSORSHIP: TNO Nutrition and Food Research was assigned by Roquette Freres to perform the study.

J Clin Gastroenterol, 2004 Jul, 38(6 Suppl), S70 - 5
Prebiotics promote good health: the basis, the potential, and the emerging evidence; Van Loo JA; BACKGROUND: The prebiotics concept, which was launched in 1995, concerns nondigested and selectively fermented carbohydrate food ingredients . It was thought that their effect in the colon could reduce risk for disease . The prebiotic concept is revisited and possible mechanisms are proposed . The physiologic consequences of prebiotic consumption are evaluated in terms of potential to reduce risk for disease . This is a compilation of several research papers, each of which complied with the World Medical Association Declaration of Helsinki . METHODS: For human dietary intervention trials, the aim was to perform double-blind, placebo-controlled, cross-over studies . A parallel design was used only for long-term studies . Most research has been done with beta(2-1) fructans, so they are used as an example of prebiotics here . RESULTS: The results are relevant to the fields of gut function, lipid metabolism, mineral absorption, bone formation, immunology, and cancer . CONCLUSION: It is observed that modification of intestinal flora by inherently selectively fermented prebiotics is central in determining their nutritional properties . They interact positively through the large intestinal surface with various physiologic processes and are thought to improve health status by reducing risk for disease (markers).

J Clin Gastroenterol, 2004 Jul, 38(6 Suppl), S67 - 9
Tolerance of probiotics and prebiotics; Marteau P et al.; The clinical efficacy of probiotics and prebiotics has been proved in several clinical settings . The authors review their proved or potential side effects . Probiotics as living microorganisms may theoretically be responsible for 4 types of side effects in susceptible individuals: infections, deleterious metabolic activities, excessive immune stimulation, and gene transfer . Very few cases of infection have been observed . These occurred mainly in very sick patients who received probiotic drugs because of severe medical conditions . Prebiotics exert an osmotic effect in the intestinal lumen and are fermented in the colon . They may induce gaseousness and bloating . Abdominal pain and diarrhea only occur with large doses . An increase in gastroesophageal reflux has recently been associated with large daily doses . Tolerance depends on the dose and individual sensitivity factors (probably the presence of irritable bowel syndrome or gastroesophageal reflux), and may be an adaptation to chronic consumption.

J Nat Prod, 2004 Jun, 67(6), 1036 - 8
Isolation and structure of antagonists of chemokine receptor (CCR5); Jayasuriya H et al.; Human CCR5 is a G-coupled receptor that binds to the envelope protein gp120 and CD4 and mediates the HIV-1 viral entry into the cells . The blockade of this binding by a small molecule receptor antagonist could lead to a new mode of action agent for HIV-1 and AIDS . Screening of natural product extracts led to the identification of anibamine (1), a novel pyridine quaternary alkaloid as a TFA salt, from Aniba sp.; ophiobolin C from fermentation extracts of fungi Mollisia sp.; and 19,20-epoxycytochalasin Q from Xylaria sp . Formation of the TFA salt of anibamine is plausibly an artifact of the isolation . The identity of the natural counterion is unknown . Anibamine.TFA competed for the binding of 125I-gp120 to human CCR5 with an IC50 of 1 microM . Ophiobolin C and 19,20-epoxycytochalasin Q exhibited binding IC50) values of 40 and 60 microM, respectively.

J Antibiot (Tokyo), 2004 Apr, 57(4), 264 - 70
FR901512, a novel HMG-CoA reductase inhibitor produced by an agonomycetous fungus No . 14919 . II . Taxonomy of the producing organism, fermentation, isolation and physico-chemical properties; Hatori H et al.; Novel compounds FR901512 and FR901516 were isolated from the fermentation broth of agonomycete strain No . 14919 . FR901512 and FR901516 possess unique tetralin ring in their structure . These compounds were potent inhibitors of the cholesterol synthesis in human hepatoma cell line Hep G2 . FR901512 shows strong 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitory activity with an IC50 value of 0.95 nM.

Adv Biochem Eng Biotechnol, 2004, 89, 197 - 223
The application of multi-parameter flow cytometry to monitor individual microbial cell physiological state; Hewitt CJ et al.; The development of multi-parameter flow cytometric techniques in our laboratories has led to a functional classification of the physiological state of single celled micro-organisms, including both yeast and bacteria . This classification is based on the presence or absence of an intact fully polarized cytoplasmic membrane and the transport systems across it . Using these techniques it is possible to resolve a cells physiological state, beyond culturability to include metabolic activity enabling assessment of population heterogeneity . Importantly results are available in real-time, 1-2 min after a sample is taken, enabling informed decisions to be taken about a process . These techniques have been extensively applied by us for monitoring the stress responses of micro-organisms in such diverse areas as brewing, bio-remediation, bio-transformation, food processing and pharmaceutical fermentation, some of which are discussed here.

Adv Biochem Eng Biotechnol, 2004, 89, 93 - 142
Inclusion bodies: formation and utilisation; Fahnert B et al.; The efficient in vivo folding of many heterologous proteins is a major bottleneck of high level production in bacterial hosts and simple optimisation protocols have not been available yet . Therefore, inclusion body (IB) based processes play a major role as a potential strategy for the production of complex recombinant proteins . These processes combine the advantages of a high accumulation of the target protein in well-characterised bacteria such as Escherichia coli, efficient strategies for IB isolation, purification and in vitro protein refolding without the need of complicated coexpression systems . Recent advances in the molecular physiology of IB formation and resolubilisation allow straight-forward optimisation of fermentation processes to obtain a high-quality product . In addition, simple strategies have been developed to optimise the purification and renaturation of disulfide bond containing proteins making a fast transfer of such processes into the industrial production scale realistic.

Adv Biochem Eng Biotechnol, 2004, 87, 195 - 268
Developing a sustainable bioprocessing strategy based on a generic feedstock; Webb C et al.; Based on current average yields of wheat per hectare and the saccharide content of wheat grain, it is feasible to produce wheat-based alternatives to many petrochemicals . However, the requirements in terms of wheat utilization would be equivalent to 82% of current production if intermediates and primary building blocks such as ethylene, propylene, and butadiene were to be produced in addition to conventional bioproducts . If only intermediates and bioproducts were produced this requirement would fall to just 11%, while bioproducts alone would require only 7% . These requirements would be easily met if the global wheat yield per hectare of cultivated land was increased from the current average of 2.7 to 5.5 tonnes ha(-1) (well below the current maximum) . Preliminary economic evaluation taking into account only raw material costs demonstrated that the use of wheat as a generic feedstock could be advantageous in the case of bioproducts and specific intermediate petrochemicals . Gluten plays a significant role considering the revenue occurring when it is sold as a by-product . A process leading to the production of a generic fermentation feedstock from wheat has been devised and evaluated in terms of efficiency and economics . This feedstock aims at providing a replacement for conventional fermentation media and petrochemical feedstocks . The process can be divided into four major stages--wheat milling; fermentation of whole wheat flour by A . awamori leading to the production of enzymes and fungal cells; glucose enhancement via enzymatic hydrolysis of flour suspensions; and nitrogen/micronutrient enhancement via fungal cell autolysis . Preliminary costings show that the operating cost of the process depends on plant capacity, cereal market price, presence and market value of added-value by-products, labour costs, and mode of processing (batch or continuous).

Adv Biochem Eng Biotechnol, 2004, 87, 151 - 71
Industrial application of fuzzy control in bioprocesses; Honda H et al.; In a bioprocess, for example a fermentation process, many biological reactions are always working in intracellular space and the control of such a process is very complicated . Bioprocesses have therefore been controlled by the judgment of the experts who are the skilled operators and have much experience in the control of such processes . Such experience is normally described in terms of linguistic IF-THEN rules . Fuzzy inference is a powerful tool for incorporating linguistic rules into computer control of such processes . Fuzzy control is divided into two types--direct fuzzy control of process variables, for example sugar feed rate and fermentation temperature, and indirect control via phase recognition . In bioprocess control the experts decide the value of controllable process variables such as sugar feed rate or temperature as output data from several state variables as input data . Fuzzy control is regarded as a computational algorithm in which the causal relationship between input and output data are incorporated . In Japan fuzzy control has already been applied to practical industrial processes such as production of pravastatin precursor and vitamin B2 and to the Japanese sake mashing process; these examples are reviewed . In addition, an advanced control tool developed from a study on fuzzy control, fuzzy neural networks (FNN), are introduced . FNN can involve complicated causality between input and output data in a network model . FNN have been proven to be applicable to a research in biomedicine, for example modeling of the complicated causality between electroencephalogram or gene expression profiling data and prognostic prediction . Successful results on this research will be also explained.

J Anim Sci, 2004 Jun, 82(6), 1847 - 54
Effects of a twin strain of saccharomyces cerevisiae live cells on mixed ruminal microorganism fermentation in vitro; Lila ZA et al.; This experiment was designed to investigate the effects of different concentrations (0, 0.33, 0.66, 0.99, and 1.32 g/L) of a twin-strain of Saccharomyces cerevisiae live cells on in vitro mixed ruminal microorganism fermentation of corn starch, soluble potato starch, and sudangrass hay (60.5%, DM basis) plus concentrate mixture (39.5%, DM basis) . Ruminal fluid was collected from two dairy cows, mixed with phosphate buffer (1:2), and incubated (30 mL) anaerobically at 38 degrees C for 6 and 24 h with or without yeast supplement, using 200 mg (DM basis) of each substrate . Medium pH, ammonia-N, and numbers of protozoa were unaffected (P = 0.38) by yeast cells in all substrates . Molar proportion of acetate was unchanged (P = 0.56) with cornstarch and soluble potato starch, but increased quadratically (P = 0.02) with hay plus concentrate by treatment . Addition of yeast cells caused a linear increase of total VFA (P = 0.008) in all substrates . Excluding the soluble potato starch, supplementation of S . cerevisiae resulted in a quadratic increase of propionate (P = 0.01), with a quadratic decrease (P = 0.04) of acetate:propionate . When soluble potato starch was used as a substrate, a linear increase (P = 0.006) of the molar proportion of propionate and a quadratic decrease (P = 0.007) in acetate:propionate was observed by treatment . Molar proportion of butyrate was unchanged (P = 0.35) with cornstarch and soluble potato starch, whereas it decreased linearly (P = 0.007) with hay plus concentrate by yeast cell supplementation . When cornstarch and soluble potato starch were used as a substrate, minor VFA were decreased (P = 0.05) by treatment . Accumulation of lactate was linearly decreased by treatment (P = 0.007) in all substrates . During incubation with hay plus concentrate, IVDMD was linearly increased (P = 0.006), whereas production of methane (linear; P = 0.02) and accumulation of hydrogen was decreased (quadratic; P = 0.005) by treatment after 24 h . These results showed that a twin strain of S . cerevisiae live cells stimulated in vitro mixed ruminal microorganism fermentation with decreased lactate, and a small decrease of methane and hydrogen with hay plus concentrate.

J Anim Sci, 2004 Jun, 82(6), 1839 - 46
Effect of Japanese horseradish oil on methane production and ruminal fermentation in vitro and in steers; Mohammed N et al.; The effects of alpha-cyclodextrin-horseradish oil complex (CD-HR) on methane production and ruminal fermentation were studied in vitro and in steers . In the in vitro study, diluted ruminal fluid (30 mL) was incubated anaerobically at 38 degrees C for 6 h with or without CD-HR, using cornstarch as substrate . The CD-HR was added at various concentrations (0, 0.17, 0.85 and 1.7 g/L) . Treatment affected neither the pH of the medium nor the number of protozoa . Total VFA increased in a linear manner (P = 0.02), and NH3-N decreased quadratically (P = 0.04) as the concentration of CD-HR increased from 0.17 g/L to 1.7 g/L . Molar proportions of acetate decreased in a linear manner (P = 0.03), and propionate increased linearly (P = 0.008) with increasing concentrations of CD-HR . Production of methane was inhibited up to 90%, whereas accumulation of dihydrogen was increased 36-fold by 1.7 g/L of CD-HR supplementation relative to controls . The effect of CD-HR on methane production, ruminal fermentation and microbes, and digestibility was further investigated in vivo using four Holstein steers in a crossover design . The CD-HR supplement was mixed into the concentrate portion of a (1.5:1) Sudangrass hay plus concentrate mixture that was fed twice daily to the steers . Ruminal samples were collected 0, 2, and 5 h after the morning feeding . No effects of CD-HR supplementation on ruminal pH (P = 0.63) or protozoal numbers (P = 0.44) were observed . Molar proportion of acetate was decreased (P = 0.04) and propionate was increased (P = 0.005) by CD-HR treatment . Molar proportion of butyrate was increased (P = 0.05) in CD-HR-supplemented steers . Ruminal NH3-N was decreased (P = 0.05) by treatment . Blood plasma glucose concentration was increased (P = 0.02) and urea-N was decreased (P = 0.04) with CD-HR supplementation . Daily DMI was decreased (P = 0.04), and apparent digestibility of DM (P = 0.13), NDF (P = 0.14), and CP tended (P = 0.14) to be increased by treatment . Methane production was decreased (P = 0.03) by 19%, and the number of methanogens was also decreased (P = 0.03) . Although N retention (P = 0.11), total viable bacteria (P = 0.15), and sulfate-reducing bacteria (P = 0.17) were not significantly altered by treatment, tendencies for increases were noted with CD-HR supplementation . The number of cellulolytic (P = 0.38) and acetogenic bacteria (P = 0.32) remained unchanged by treatment . These results indicate that CD-HR supplementation can be used to decrease methane production in steers.

J Anim Sci, 2004 Jun, 82(6), 1818 - 25
Effect of imbalance between energy and nitrogen supplies on microbial protein synthesis and nitrogen metabolism in growing double-muscled Belgian Blue bulls; Valkeners D et al.; Six double-muscled Belgian Blue bulls (initial weight: 345 +/- 16 kg) with cannulas in the rumen and proximal duodenum were used in two juxtaposed 3 x 3 Latin squares to study the effect of a lack of synchronization between energy and N in the rumen on microbial protein synthesis and N metabolism by giving the same diet according to three different feeding patterns . The feed ingredients of the diet were separated into two groups supplying the same amount of fermentable OM (FOM), but characterized by different levels of ruminally degradable N (RDN) . The first group primarily provided energy for the ruminal microbes (14.6 g of RDN/kg of FOM), and the second provided N (33.3 g of RDN/kg of FOM) . These two groups were fed to the bulls simultaneously or alternately with the aim of creating three different time periods of imbalance (0, 12, or 24 h) between energy and N supplies in the rumen . The introduction of imbalance affected neither microbial-N flow at the duodenum (P = 0.65) nor efficiency of growth (P = 0.69), but decreased (P = 0.016) the NDF degradation in the rumen 12.2% for a 12-h period of imbalance . N retention was not affected by imbalance (P = 0.53) and reached 57.8, 58.5, and 54.7 g/d, respectively, for 0-, 12- and 24-h imbalance . It seems that the introduction of an imbalance of 12 or 24 h between energy and N supplies for the ruminal microbes by altering the feeding pattern of the same diet does not negatively influence microbial protein synthesis or N retention by the animal . Nitrogen recycling in the rumen plays a major role in regulating the amount ofruminally available N and allows for continuous synchronization of N- and energy-yielding substrates for the microorganisms in the rumen . Therefore, a lack of synchronization in the diet between the energy and N supplies for the ruminal microbes is not detrimental to their growth or for the animal as long as the nutrient supply is balanced on a 48-h basis . Thus, these dietary feeding patterns may be used under practical feeding conditions with minimal effect on the performance of ruminant animals.

Lett Appl Microbiol, 2004, 38(4), 315 - 20
Microsatellite PCR profiling of Saccharomyces cerevisiae strains during wine fermentation; Howell KS et al.; AIMS: Use of microsatellite PCR to monitor populations of Saccharomyces cerevisiae strains during fermentation of grape juice . METHOD AND RESULTS: Six commercial wine strains of S . cerevisiae were screened for polymorphism at the SC8132X locus using a modified rapid PCR identification technique . The strains formed four distinct polymorphic groups that could be readily distinguished from one another . Fermentations inoculated with mixtures of three strains polymorphic at the SC8132X locus were monitored until sugar utilization was complete, and all exhibited a changing population structure throughout the fermentation . CONCLUSIONS: Rapid population quantification demonstrated that wine fermentations are dynamic and do not necessarily reflect the initial yeast population structure . One or more yeast strains were found to dominate at different stages of the fermentation . SIGNIFICANCE AND IMPACT OF THE STUDY: The population structure of S . cerevisiae during mixed culture wine fermentation is dynamic and could modify the chemical composition and flavour profile of wine.

Environ Technol, 2004 Apr, 25(4), 391 - 401
Impact of long chain fatty acids on glucose fermentation under mesophilic conditions; Lalman JA et al.; The effects of three long chain fatty acids (LCFAs) on glucose fermentation at 37 degrees C were examined during this study . Linoleic acid (LA) was more inhibitory than oleic acid (OA) and stearic acid (SA) . During glucose fermentation, the carbon flow was diverted towards the production of more reduced volatile fatty acids (VFAs) in cultures fed with unsaturated LCFAs . In cultures inoculated with LA and OA, butyrate was produced with elevated levels observed in cultures inoculated with LA . Propionate degradation was unaffected by the presence of SA; however, elevated levels and longer removal times were observed in cultures receiving LA and OA . Acetate accumulation indicated the acetoclastic methanogenic population was affected only by OA and LA.

Am J Clin Nutr, 2004 Jul, 80(1), 89 - 94
L-Rhamnose increases serum propionate in humans; Vogt JA et al.; BACKGROUND: Acetic and propionic acids are produced by colonic bacterial fermentation of unabsorbed carbohydrates and are absorbed into the portal circulation . From there, they travel to the liver, where acetate is a lipogenic substrate and propionate can inhibit lipogenesis . The extent to which peripheral blood short-chain fatty acid concentrations reflect differences in colonic fermentation is uncertain . The unabsorbed sugar lactulose produces mainly acetate when fermented in vitro, whereas L-rhamnose yields propionate . OBJECTIVE: The objective of the study was to ascertain whether ingestion of L-rhamnose and lactulose would have different acute effects on peripheral acetate and propionate concentrations and on breath hydrogen and methane concentrations . DESIGN: Twenty-two subjects were fed 25 g L-rhamnose, lactulose, or glucose on 3 separate occasions in a randomized crossover design . Blood and breath samples were collected hourly for 12 h . RESULTS: Serum propionate was significantly higher with ingestion of L-rhamnose than with that of lactulose or glucose (P < 0.001) . The area under the curve for serum acetate was significantly higher with ingestion of lactulose than with that of glucose (P < 0.03) . The ratio of serum acetate to propionate was significantly higher with ingestion of lactulose than with that of glucose or L-rhamnose (P < 0.01) . Breath hydrogen was significantly higher with ingestion of lactulose than with that of L-rhamnose or glucose (P < 0.0001) . CONCLUSIONS: The selective increases in serum acetate and propionate concentrations in humans were obtained by feeding specific fermentable substrates . Presumably, these changes in serum concentrations reflect changes in colonic production . Selective alteration of colonic fermentation products could yield a new mechanism for modifying blood lipids.

J Agric Food Chem, 2004 Jun 30, 52(13), 4172 - 7
Manufacture of fermentable sugar solutions from sugar cane bagasse hydrolyzed with phosphoric acid at atmospheric pressure; Gamez S et al.; Sugar cane bagasse, a renewable and cheap bioresource, was hydrolyzed at 100 degrees C using phosphoric acid at different concentrations (2, 4, or 6%) and reaction times (0-300 min) to obtain fermentable sugar solutions, which have a high concentration of sugars (carbon source for microorganism growth) and a low concentration of growth inhibitors (acetic acid and furfural) . Xylose, glucose, arabinose, acetic acid, and furfural were determined following the hydrolysis . Kinetic parameters of mathematical models for predicting these compounds in the hydrolysates were obtained . Derived parameters such as efficiency of hydrolysis or purity of hydrolysates were considered to select as optimal conditions 6% phosphoric acid at 100 degrees C for 300 min . Using these conditions, 21.4 g of sugars L(-)(1) and <4 g of inhibitors L(-)(1) were obtained from the hydrolysis with a water/solid ratio of 8 g of water g(-)(1) of sugar cane bagasse on a dry basis.

Nutr Hosp, 2004 May-Jun, 19(3), 167 - 74
{Fiber in enteral nutrition: systematic review of the literature}; del Olmo D et al.; It has been suggested that adding fibre to enteral nutrition (EN) formulas may achieve a normalization of the gastrointestinal tract's functions by reducing the incidence of diarrhoea and constipation . However, this supposition is based on the demonstrated effect of fibre in natural feeding . There have so far been no firm recommendations on the use, amount or type of fibre that is most advisable in EN . GOAL: To establish the scientific evidence existing on the benefits gained by adding fibre to EN formulas compared with the use of fibre-free formulas . MATERIAL AND METHODS: Bibliographical search on PubMed, on the register of Randomized Prospective Trials (RPT) of the Cochrane Library and manual search . Inclusion criteria: RPT comparing fibre-free formulas with isocaloric ad isonitrogen formulas containing fibre in total EN . Four independent reviewers revised the references selected . The data were analyzed using the RevMan 4.1 programme from the Cochrane Library . RESULTS: Of 286 references, 25 fulfilled the inclusion criteria . The data were grouped by population and outcome variable . The addition of fermentable fibre to the EN formulas tends to reduce the incidence of diarrhoea in critically-ill and post-surgery patients (OR = 0.66; 95% CI 0.46-0.95; p = 0.19) . The use of formulas with insoluble fibre seems to increase the frequency of depositions and reduce the need for laxatives in immobilized patients receiving long-term EN but the data are insufficient . In people with normal gastrointestinal functions, the frequency of depositions is similar when using formulas with and without fibre (DPM = -0.97; 95% CI -0.12-{-0.02}; p < 0.0001) . CONCLUSION: There is currently insufficient evidence to recommend the systematic use of formulas containing fibre in total EN . However, in critically-ill and post-surgery patients, the use of formulas with fermentable fibre tends to reduce the incidence of diarrhoea . Although information is still lacking . The formulas with insoluble fibre seem to diminish constipation in chronic EN, whereas in other situations where the digestive system is working appropriately, no benefit has been shown . It is necessary to increase the number and quality of these trials in order to be able to establish firm recommendations on the use of EN formulas containing fibre.

Biotechnol Bioeng, 2004 Jul 5, 87(1), 90 - 8
Fermentation performance of engineered and evolved xylose-fermenting Saccharomyces cerevisiae strains; Sonderegger M et al.; Lignocellulose hydrolysate is an abundant substrate for bioethanol production . The ideal microorganism for such a fermentation process should combine rapid and efficient conversion of the available carbon sources to ethanol with high tolerance to ethanol and to inhibitory components in the hydrolysate . A particular biological problem are the pentoses, which are not naturally metabolized by the main industrial ethanol producer Saccharomyces cerevisiae . Several recombinant, mutated, and evolved xylose fermenting S . cerevisiae strains have been developed recently . We compare here the fermentation performance and robustness of eight recombinant strains and two evolved populations on glucose/xylose mixtures in defined and lignocellulose hydrolysate-containing medium . Generally, the polyploid industrial strains depleted xylose faster and were more resistant to the hydrolysate than the laboratory strains . The industrial strains accumulated, however, up to 30% more xylitol and therefore produced less ethanol than the haploid strains . The three most attractive strains were the mutated and selected, extremely rapid xylose consumer TMB3400, the evolved C5 strain with the highest achieved ethanol titer, and the engineered industrial F12 strain with by far the highest robustness to the lignocellulosic hydrolysate .

Biotechnol Bioeng, 2004 Jul 5, 87(1), 54 - 60
Zeta potential as a measure of polyelectrolyte flocculation and the effect of polymer dosing conditions on cell removal from fermentation broth; Pearson CR et al.; Characterization of flocculation for cell removal from fermentation broth via polyelectrolyte addition is commonly based on qualitative methods such as physical appearance of the floc . The use of zeta potential as a quantitative measure of floc character was evaluated as an indicator of optimal polymer addition . Zeta potential was found to increase with increasing cationic polyelectrolyte dosage, but never reached zero regardless of the total amount of polymer added, indicating flocculation occurs at least partially through a bridging type mechanism . Experiments were conducted using various polymer concentrations (25-75 g/L) and dosing methods (batch, incremental and continuous addition) that resulted in variable overall polymer requirements to achieve optimum flocculation . Zeta potential was found to be constant at optimal floc character regardless of the total amount of polymer added, polymer concentration, or method of polymer addition . Experiments with two additional types of fermentation broth also showed characteristic zeta potentials at optimal flocculation . Polymer requirements to achieve a particular floc character can vary greatly, depending on polymer dosing conditions and fermentation batch . The effect of polymer dosing conditions on the polymer requirement to obtain optimal floc character was evaluated . Polymer dosing method and calcium concentration were both found to have a significant effect (P < 0.0001) with continuous polymer addition and high calcium concentration requiring less polymer than did batch polymer addition and low calcium concentration, respectively . Polymer dosing concentration did not significantly affect polymer requirement for optimal flocculation .

J Biol Chem, 2004 Aug 20, 279(34), 35219 - 27 Epub 2004 Jun 21.
Cytosolic thioredoxin peroxidase I and II are important defenses of yeast against organic hydroperoxide insult: catalases and peroxiredoxins cooperate in the decomposition of H2O2 by yeast; Munhoz DC et al.; The cytosolic thioredoxin peroxidase II (cTPxII/Tsa2p) from Saccharomyces cerevisiae shares 86% identity with the relatively well characterized cytosolic thioredoxin peroxidase I (cTPxI/Tsa1p) . In contrast to cTPxI protein, cTPxII is not abundant and is highly inducible by peroxides . Here, we describe a unique phenotype for DeltacTPxII strain; these cells were highly sensitive to tert-butylhydroperoxide (TBHP) but presented resistance to H(2)O(2) in fermentative and respiratory conditions . In contrast, DeltacTPxI strain was very sensitive to both TBHP and H(2)O(2), whatever the carbon source present in the media . These differences in the response of mutant cells to the different kinds of peroxide insult could not be attributed to enzymatic properties of cTPxI and cTPxII since the recombinant proteins showed similar in vitro efficiencies (K(cat) /K(m)) in the removals of both kinds of peroxide . This specific sensitivity of DeltacTPxII cells to TBHP could not be related to the expression pattern of TSA2 (cytosolic thioredoxin peroxidase II gene) either, since this gene is highly inducible by both H(2)O(2) and TBHP when cells were grown in different conditions . Finally, peroxide-removing assays were performed and showed that catalase activity increased significantly only in DeltacTPxII cells, which appear to be related with the resistance of this strain to H(2)O(2) . Taken together, present data indicate that cTPxII and cTPxI are key components of the yeast defense system against organic peroxide insult . In regard to the stress induced by H(2)O(2), catalases (peroxisomal and/or cytosolic) and cTPxII seemed to cooperate with cTPxI in the defense of yeast against this oxidant.

Br J Cancer, 2004 May 4, 90(9), 1792 - 5
Do preserved foods increase prostate cancer risk?
Jian L, Zhang DH, Lee AH, Binns CW.
Preserved foods have been found in some studies to be associated with increased cancer risks . The possible relationship between preserved foods and prostate cancer was investigated in a case-control study in southeast China during 2001-2002 covering 130 histologically confirmed cases and 274 inpatient controls without malignant disease . The total amount of preserved food consumed was positively associated with cancer risk, the adjusted odds ratio being 7.05 (95% CI: 3.12-15.90) for the highest relative to the lowest quartile of intake . In particular, the consumption of pickled vegetables, fermented soy products, salted fish and preserved meats was associated with a significant increase in prostate cancer risk, all with a significant dose-response relationship.

Bioresour Technol, 2004 Oct, 95(1), 49 - 52
Pectinase production by Aspergillus niger using wastewater in solid state fermentation for eliciting plant disease resistance; Bai ZH et al.; An elicitor of plant disease resistance, pectinase, was produced by solid state fermentation with Aspergillus niger . Sugar beet pulp was used as carbon source and the wastewater from monosodium glutamate production was used as nitrogen and water source . The composition of the fermentation medium was: 11 ml concentrated wastewater (containing NH3-N 38.2 mg/ml), sugar beet pulp 10 g, Na2HPO4.12H2O 0.2 g, KH2PO4 0.04 g in a 500 ml Erlenmeyer flask . The fermentation temperature was 30 degrees C and the relative humidity of the air was 75-90% . The maximum production of pectinase was reached after 96 h cultivation . The crude pectinase extracted from the fermented materials could elicit disease resistance in cucumber and tomato seedlings.

Bioresour Technol, 2004 Oct, 95(1), 31 - 3
Growth of Azotobacter vinelandii in a solid-state fermentation of technical lignin; Zhang X et al.; Azotobacter vinelandii was cultured on technical lignin, derived from Kraft pulping processes, for biofertilizer production in solid-state fermentation . The effects of the ratio of technical lignin to corn straw, initial water content, and material bed depth on the microorganisms were studied in detail . At 30 degrees C, technical lignin to corn straw at the ratio of 1:0.75, the bed depth of 5 cm, and 67% moisture content, A . vinelandii was grown and reached 4.2 x 10(10) cfu g(-1) dry rot after 36 h.

Food Addit Contam, 2004 Jun, 21(6), 607 - 17
Screening survey of deoxynivalenol in beer from the European market by an enzyme-linked immunosorbent assay; Papadopoulou-Bouraoui A et al.; Deoxynivalenol (DON) was analysed in 313 beer samples collected from the European retail market using a commercially available immunoassay kit (enzyme-linked immunosorbent assay, ELISA) . The incidence rate was about 87%, while most samples (73%) had contamination levels lower than 20 ng m(-1) . The contamination ranged between 4.0 and 56.7 ng ml(-1), with an average of 13.5 ng ml(-1) . A statistically significant correlation between alcohol levels and DON contamination was found, as well as a significant difference between bottom, top and spontaneous fermenting beers . Twenty-seven beer samples were compared using a second ELISA kit and a good correlation was obtained between the two kits (r = 0.93) . Although when compared with gas chromatography-mass spectrometry the ELISA tended to overestimate the results, a good correlation (r=0.94) between the two methods was observed . Monitoring of DON in beer is important considering that DON production is dependent on the weather and that it can contribute significantly to the tolerable daily intake of DON, especially for frequent beer consumers.

Nutr Cancer, 2004, 48(1), 1 - 5
A hypothesis: interaction between supplemental iron intake and fermentation affecting the risk of colon cancer . The Iowa Women's Health Study; Lee DH et al.; Fermentation in the large intestine can increase absorption of ferrous iron, which is the main form in supplements, because the solubility of ferrous iron is enhanced in the mildly acidic environments caused by fermentation . We therefore hypothesized that higher supplemental iron intake would increase the risk of colon cancer among those who consume large amounts of fermentable substrates, namely, dietary fiber and resistant starch . Among 34,708 postmenopausal women, supplemental iron was unrelated to proximal colon cancer in all women and to distal colon cancer among those consuming below the median of fermentable substrates . However, supplemental iron was positively associated with distal colon cancer among women who consumed above the median of fermentable substrates (P for interaction %lt; 0.01); the adjusted relative risks across categories of supplemental iron (0 g/day, 1-19 g/day, 20-49 g/day, and > or = 50 g/day) were 1.0, 1.24, 1.78, and 3.78 (P for trend < 0.01) . This hypothesis needs confirmation in other cohort studies because, despite the significant trend, only nine cases were included in the top category of > or = 50 mg supplemental iron, and this finding could have arisen by chance .

Anal Biochem, 2004 Jul 15, 330(2), 311 - 6
Microarrays of peptides elevated on the protein layer for efficient protein kinase assay; Lee SJ et al.; Peptide microarrays can be used for the high-throughput analysis of protein-peptide interactions . However, current peptide microarrays are rather costly to make and require cumbersome steps of introducing novel polymeric surfaces and/or chemical derivatization of peptides . Here, we report a novel method for manufacturing peptide microarrays by elevating the peptide on the layer of protein by a fusion protein approach . Using two protein kinases and their peptide substrates as examples, we show that elevating peptides on the layer of protein allows sensitive, specific, and efficient detection of peptide-protein interactions without the need for complicated chemical modification of solid supports and peptides . It was found that kinase activity could be detected with as low as 0.09 fmol of kemptide, which is about 1000-fold more sensitive than the 0.1 pmol obtained with other microarray systems . Furthermore, peptides can be produced as fusion proteins by fermentation of recombinant Escherichia coli and thus the expensive peptide synthesis process can be avoided . Therefore, this new strategy will not only be useful in high-throughput and cost-effective screening of kinase substrate peptides but also be generally applicable in studying various protein-peptide interactions.

J Dairy Sci, 2004 Mar, 87(3), 665 - 71
Performance of lactating dairy cows fed whole cottonseed with elevated concentrations of free fatty acids in the oil; Sullivan HM et al.; Twenty-four multiparous cows were used in a 10-wk randomized block design trial to evaluate the effects of feeding whole cottonseed (WCS) containing increasing concentrations of free fatty acids (FFA) in the oil on nutrient intake and digestibility, milk yield and composition, and select plasma metabolites . Two lots of WCS containing either 3 or 12% FFA were blended to provide WCS with 3, 6, 9, and 12% FFA . Cottonseeds were included in the wheat silage-based total mixed ration at 12.5% of dry matter (DM) . There was no difference in intakes of DM, crude protein (CP), or neutral detergent fiber; yield of milk; or percentages of milk protein, lactose, or SNF . Milk fat percentage was lowest for the diet containing WCS with 6% FFA . Concentrations of individual milk fatty acids C6:0 decreased and C16:1 increased linearly as FFA in WCS increased . A cubic response was observed for concentrations of C8:0, C10:0, and C12:0 because of higher concentrations when diets contained WCS with 6% FFA than 3 and 12% FFA, which were higher than 9% FFA . Intake and apparent total tract digestibility of acid detergent fiber increased linearly as FFA concentration in WCS increased . Apparent NDF digestibility was highest for diets containing WCS with 3 and 6% FFA; CP digestibility was highest with WCS containing 3 and 9% FFA . Differences in milk fatty acid concentration and nutrient digestibility suggest minor changes in rumen fermentation; however, feeding WCS with up to 12% FFA did not negatively impact nutrient intake and digestibility or milk yield or composition.

J Dairy Sci, 2004 Mar, 87(3), 637 - 44
Comparison of brown midrib-6 and -18 forage sorghum with conventional sorghum and corn silage in diets of lactating dairy cows; Oliver AL et al.; Total mixed rations containing conventional forage sorghum, brown midrib (bmr)-6 forage sorghum, bmr-18 forage sorghum, or corn silage were fed to Holstein dairy cows to determine the effect on lactation, ruminal fermentation, and total tract nutrient digestion . Sixteen multiparous cows (4 ruminally fistulated; 124 d in milk) were assigned to 1 of 4 diets in a replicated Latin square design with 4-wk periods (21-d adaptation and 7 d of collection) . Diets consisted of 40% test silage, 10% alfalfa silage, and 50% concentrate mix (dry basis) . Acid detergent lignin concentration was reduced by 21 and 13%, respectively, for the bmr-6 and bmr-18 sorghum silages when compared with the conventional sorghum . Dry matter intake was not affected by diet . Production of 4% fat-corrected milk was greatest for cows fed bmr-6 (33.7 kg/d) and corn silage (33.3 kg/d), was least for cows fed the conventional sorghum (29.1 kg/d), and was intermediate for cows fed the bmr-18 sorghum (31.2 kg/d), which did not differ from any other diet . Total tract neutral detergent fiber (NDF) digestibility was greatest for the bmr-6 sorghum (54.4%) and corn silage (54.1%) diets and was lower for the conventional (40.8%) and bmr-18 sorghum (47.9%) diets . In situ extent of NDF digestion was greatest for the bmr-6 sorghum (76.4%) and corn silage (79.0%) diets, least for the conventional sorghum diet (70.4%), and intermediate for the bmr-18 sorghum silage diet (73.1%), which was not different from the other diets . Results of this study indicate that the bmr-6 sorghum hybrid outperformed the conventional sorghum hybrid; the bmr-18 sorghum was intermediate between conventional and bmr-6 in most cases . Additionally, the bmr-6 hybrid resulted in lactational performance equivalent to the corn hybrid used in this study . There are important compositional differences among bmr forage sorghum hybrids that need to be characterized to predict animal response accurately.

J Dairy Sci, 2004 Mar, 87(3), 630 - 6
Fermentation acids, aerobic fungal growth, and intake of napiergrass ensiled with nonfiber carbohydrates; Yang CM et al.; This study evaluated fermentation characteristics and fungal numbers of napiergrass silages prepared with and without added raw or heated corn meal (10%, fresh-weight basis) at ensiling . Corn was added to napiergrass so that the silage would contain concentrate similar to that of corn silage with minimum grain content . The silages treated with raw or heated corn were fed to dairy does to compare voluntary silage consumption . After 8 wk of fermentation, pH for silage treated with heated corn was lowest, and that for napiergrass ensiled alone was highest among the treatments . The addition of corn increased lactic acid, but propionic and butyric acids were also elevated . Acetic acid decreased in response to the supplementation of corn but remained the dominant acid for all silages . Numbers of fungi (yeasts plus molds) in silages did not differ significantly at silo opening or after 24 h of exposure to air . However, the numbers of fungi at 48 h in aerated silages containing corn were lower than were the fungi counts in the control silage . When the silages were offered free choice along with concentrate at a fixed rate, dairy does tended to consume more silage treated with heated corn than raw corn . Whole-tract nutrient digestibility and serum glucose and urea N were not altered . These results indicate that the addition of either raw or heated corn meal to napiergrass at ensiling was beneficial to silage fermentation quality because it decreased pH, increased lactic acid, and apparently suppressed fungal populations via elevated antifungal acids . A further advantage of heated corn vs . raw corn was increased silage intake.

AAPS PharmSciTech . 2003 Oct 21;4(4):E57.
Investigations on neomycin production with immobilized cells of Streptomyces marinensis NUV-5 in calcium alginate matrix; Srinivasulu B et al.; The purpose of this investigation was to study the effect of Streptomyces marinensis NUV-5 cells immobilized in calcium alginate for the production of neomycin . The effect of various parameters, such as the effect of alginate concentration (1%, 2%, 3%, 4%, and 5% wt/vol), the effect of cation (CaCl2, BaCl2, and SrCl2), the concentration of cation (0.01M, 0.125M, 0.25M, 0.375M, and 0.5M), the curing times (1, 6, 11, 16, and 21 hours), and the diameter of the bead (1.48, 2.16, 3.24, 4.46, and 5.44 mm), on neomycin production and bead stability were studied . The effect of maltose (4%, 3%, 2%, and 1% wt/vol) and sodium glutamate (0.6%, 0.3%, 0.15%, and 0.075% wt/vol) concentration on neomycin production was also studied . Better neomycin production was achieved with optimized parameters, such as alginate at 2% wt/vol, 0.25M CaCl2, 1-hour curing time, and 3.24 mm bead diameter . Effective neomycin production was achieved with 3% wt/vol maltose and 0.6% wt/vol sodium glutamate concentration . The repeated batch fermentations were conducted (every 96 hours) using the optimized alginate beads, employing the production medium with 3% wt/vol maltose and 0.6% wt/vol sodium glutamate along with mineral salts solution . The increase in antibiotic production was observed up to the 5th cycle, and later gradual decrease in antibiotic production was observed . Comparison of the total antibiotic production with free cells and immobilized cells was also done . An enhanced antibiotic productivity of 32% was achieved with immobilized cells over the conventional free-cell fermentation, while 108% more productivity was achieved over the washed free-cell fermentation . From these results it is concluded that the immobilized cells of S marinensis NUV-5 in calcium alginate are more efficient for the production of neomycin with repeated batch fermentation.

Yeast, 2004 Jun, 21(8), 671 - 84
Characterization of the effectiveness of hexose transporters for transporting xylose during glucose and xylose co-fermentation by a recombinant Saccharomyces yeast; Sedlak M et al.; We have developed recombinant Saccharomyces yeasts that can effectively co-ferment glucose and xylose to ethanol . However, these yeasts still ferment glucose more efficiently than xylose . The transport of xylose could be one of the steps limiting the fermentation of xylose . In this study, we characterized the changes in the expression pattern of the hexose transporter and related genes during co-fermentation of glucose and xylose using one of our recombinant yeasts, Saccharomyces cerevisiae 424A(LNH-ST) . The transcription of the hexose transporter and related genes was strongly influenced by the presence of glucose; HXT1, HXT2 and HXT3 were greatly activated by glucose and HXT5, HXT7 and AGT1 were significantly repressed by glucose . We also examined the effectiveness of individual transporters encoded by HXT1, HXT2, HXT4, HXT5, HXT7 and GAL2 genes for transporting xylose during co-fermentation of glucose and xylose in a Saccharomyces hxt degrees mutant (RE700A) . We compared these hxt degrees derivatives to RE700A wild-type strain (S . cerevisiae MC996A) where all of them contained the same xylose metabolizing genes present in our xylose-fermenting yeasts such as 424A(LNH-ST) . Our results showed that recombinant RE700A containing the cloned HXT7 or HXT5 were substantially more effective for fermenting xylose to ethanol . In addition, we found that the efficiency of transporters for intracellular accumulation of xylose was as follows: HXT7 > HXT5 > GAL2 > WT > HXT1 > HXT4 > > > RE700A . Furthermore, we provided evidence that the Saccharomyces galactose transporter system could be a highly effective xylose transporter . The information reported here should be of great importance for improving the Saccharomyces yeast transport of xylose .

Arch Anim Nutr, 2004 Apr, 58(2), 149 - 56
The influence of 5% and 10% dietary apple pectin on parameters of fermentation in faeces and caecal digesta of weaning pigs; Zacharias B et al.; In order to determine the effect of pectin on fermentation parameters in the faeces and caecal digesta of weaned pigs 18 castrated male crossbred pigs with an average body weight of 8 kg were fitted with T-cannulas at the caecum . The animals were randomly distributed into three groups and fed with diets supplemented with 0, 5 and 10% pectin . Faeces were collected over a period of 3 days . Thereafter the diets were withdrawn for 24 h followed by ad libitum feeding to enhance the feed intake . Caecal chyme was collected 0, 8 and 24 h postprandial . In the faeces the addition of 5% pectin to the diet lowered the content of dry matter and lactic acid . The pH and the digestibility of pectins, the concentration of total SCFA, acetate, propionate, butyrate, bicarbonate and chloride increased . Dietary pectin of 10% increased the content of total SCFA and acetate further . When the diets were withdrawn and fed ad libitum 24 h later, a decline of the pH and an increased concentration of lactate in the caecal chyme could be observed in all groups up to 8 h after feeding . With an interval of 8 to 24 h after feeding, a further decline in pH and a rise of lactate only occurred when the diet was not supplemented with pectin . It was concluded that pectin might be beneficial for the development of fermentative processes in the large intestine.

Int J Food Microbiol, 2004 Jul 15, 94(2), 211 - 5
Aflatoxins in pozol, a nixtamalized, maize-based food; Mendez-Albores JA et al.; To determine whether pozol, a nixtamalized maize-based food was contaminated with aflatoxins, samples of non-fermented pozol were collected during the period November 2002 to April 2003 from local markets at Comitan in Chiapas, Mexico . The samples were analyzed for the presence of aflatoxins . Nineteen out of one hundred and eleven samples were contaminated with aflatoxin B2 (AFB2) and traces of aflatoxin B1 (AFB1) . The percentage of samples contaminated with AFB2 in pozol prepared with white maize was 5.4% . Pozol mixed with toasted cacao paste had a contamination rate of 41.5% . No aflatoxins were detected in pozol prepared with yellow maize . It was found that only 1 of 19 contaminated samples had aflatoxin concentrations above 20 ppb.

Int J Food Microbiol, 2004 Jul 15, 94(2), 161 - 7
Solid substrate production of Epicoccum nigrum conidia for biological control of brown rot on stone fruits; Larena I et al.; Production of conidia of Epicoccum nigrum, a biocontrol agent of the fungal pathogen Monilinia laxa, was tested in liquid- and solid-state fermentation . Liquid fermentation was conducted in 250 ml Erlenmeyer flasks containing 50 ml of a mineral medium (containing per litre: 20 g lactose, 10 g NO3K, 1 g K2HPO4, 0.5 g MgSO4.7H2O, and 1 ml of a minor-element solution), inoculated with 2 x 10(5) E . nigrum conidia ml(-1), and incubated at 20-25 degrees C and 150 rpm for 7 days . Solid-state fermentation was carried out in specially designed plastic bags (600 cm3) (VALMIC) containing either 50 g of peat/vermiculite (1:1, w/w), or 50 g of peat/vermiculite/lentil meal (1:1:1, w/w/w) with 40% (v/w) initial moisture content . Substrate was inoculated with a conidial suspension of E . nigrum to give 10(5) conidia g(-1) substrate, and bags were incubated at 20-25 degrees C for 7 days in darkness . The amount of conidia of E . nigrum obtained in solid-state fermentation with substrate based on peat/vermiculite/lentil meal was 10-fold higher than with substrate based on peat/vermiculite or in liquid fermentation . Conidial production under these conditions was maintained in the range of 10(8) conidia g(-1) substrate from 10 to 150 days after inoculation . Germinability of these conidia was >90% . Addition of other nutrients than lentil meal to peat/vermiculite did not enhance production of conidia . Presence of peat in the substrate was necessary for good conidia production, but change in the kind of peat or vermiculite did not improve conidial production . Conidial production was similar when the substrate was inoculated with 10(5), 10(6) or 10(7) conidia g(-1) dry substrate . Incubation of bags in light conditions did not enhance conidial production . Fresh conidia produced in this solid-state fermentation system reduced the incidence and lesion diameter induced by M . laxa on peaches.

Med Hypotheses, 2004, 63(1), 42 - 6
Post-traumatic inflammatory response: perhaps a succession of phases with a nutritional purpose; Aller MA et al.; Post-traumatic inflammatory response, whether this be local or systemic, is considered to be the succession of three functional phases called nervous, immune and endocrine, that could have a nutritional significance . In the nervous phase, ischemia-reperfusion, which causes interstitial and cellular edema, is produced . Both types of edema could represent an ancestral mechanism to feed the cells by diffusion . During the immune phase, the tissues are infiltrated by inflammatory cells and bacteria . Then, extracellular digestion, by enzyme release (fermentation), and intracellular digestion by phagocytosis could be associated with a hypothetical trophic capacity for the neighbouring cells . Finally, in the late or endocrine phase nutrition mediated by the blood capillaries is established . In these three successive phases the inflammatory response goes on from an anaerobic metabolism (ischemia) through a metabolism characterized by a defective oxygen use (reperfusion, oxidative burst and heat hyperproduction) to an oxidative metabolism (oxidative phosphorilation) with a correct use of oxygen to produce usable energy . This type of metabolism is characterized by a large production of ATP, which is used to drive specialized multiple cellular processes . Since the nervous, immune and endocrine phases of the inflammatory response go from ischemia to the development of an oxidative metabolism, It is also tempting to speculate on whether the body reproduces the successive stages by which life passes from its origin without oxygen until it develops an effective, although costly, system for the use of oxygen every time we suffer post-traumatic acute inflammation.

Clin Microbiol Infect, 2004 Jun, 10(6), 576 - 8
A family outbreak of haemolytic uraemic syndrome and haemorrhagic colitis caused by verocytotoxigenic Escherichia coli O157 from unpasteurised cow's milk in Slovakia; Liptakova A et al.; This report describes a family outbreak of verocytotoxigenic Escherichia coli O157 (VTEC) infection, involving nine persons from one extended family, which occurred in eastern Slovakia . Three children suffered from haemolytic uraemic syndrome, two children had bloody diarrhoea, and four adults were asymptomatic carriers . Fourteen sorbitol-non-fermenting E . coli O157 isolates harbouring the vtx2, eae and ehxA genes were obtained . Verocytotoxin 2 activity was demonstrated in all 14 isolates . After epidemiological surveillance, the source of infection was identified as unpasteurised cow's milk.

J Anim Physiol Anim Nutr (Berl), 2004 Jun, 88(5-6), 188 - 95
Inhibition of ruminal microbial methane production by beta-cyclodextrin iodopropane, malate and their combination in vitro; Mohammed N et al.; The objective of this study was to evaluate the effects of different concentrations of l-malate (0, 5, 10 and 20 mm), 2-iodopropane-beta-cyclodextrin complex (CD-IP) (0, 0.1, 0.2 and 0.4 mm) and a combination of malate (10 and 20 mm) plus CD-IP (0.2 and 0.4 mm) on methane production from corn starch . Ruminal fluid was collected from dairy cows, mixed with phosphate buffer (1 : 2) and incubated (30 ml) anaerobically at 38 degrees C for 6 h with or without additives . Fermentation of corn starch in the presence of malate resulted in an increase (p < 0.05) in pH of the medium, total volatile fatty acid (VFA), total gas production and molar proportion of propionate . Acetate and ammonia-N concentration were unchanged . Methane production was decreased (p < 0.05) (15.5 to 20.4%) . Addition of CD-IP in corn starch resulted in an increase (p < 0.05) in total VFA and molar proportion of propionate . Acetate, pH and ammonia-N concentration of the medium were decreased (p < 0.05) . Total gas production was unchanged . Methane production was decreased (p < 0.05) (25.2 to 97.1%) and hydrogen production was increased (p < 0.05) . Addition of l-malate to CD-IP resulted in an increase (p < 0.05) in total VFA, total gas production and molar proportion of propionate . Acetate and ammonia-N concentration were decreased (p < 0.05) . No effects were observed on medium pH . Methane production was decreased (p < 0.05) (49.5 to 97.1%) . Hydrogen production was also decreased (p < 0.05) (54.5 to 64.1%) compared with those of CD-IP alone . Therefore, these additives may be used as supplements to inhibit methane production as well as to improve rumen fermentation and animal performance .

Lett Appl Microbiol, 2004, 39(1), 48 - 54
Typical metabolic traits of two Oenococcus oeni strains isolated from Valpolicella wines; Zapparoli G et al.; AIMS: Physiological comparison of two indigenous Oenococcus oeni strains, U1 and F3 isolated in the same area (Valpolicella, Italy) in order to select a performant starter for MLF in wine . METHODS AND RESULTS: Growth rate, sugar and malate metabolism in FT80 media at pH 5.3 and 3.5 were analysed . The amount of total protein synthesized and the level of expression of the small Hsp Lo18 were evaluated by radiolabelling and immunodetection experiments after heat (42 degrees C), acid (pH 3.5) and ethanol (12% v/v) stresses . Strain U1 showed significantly lower specific growth rate and growth yield in acid conditions than strain F3 . However, strain U1 had a higher malate consumption capacity at pH 3.5 than strain F3, in relation with an higher malolactic activity determined on whole cells . Strain U1 exhibited about half the total protein synthesis level than strain F3, but both strains expressed Lo18 similarly . Evaluation of malolactic fermentation (MLF) performance by microvinification trials was carried out . Strain U1 was able to complete MLF, whereas strain F3 degraded malic acid partially when inoculated in Amarone wine . CONCLUSIONS: Considering its performances in microvinifications experiments, strain U1 could be a good candidate for malolactic starter as an alternative to deficient commercial starters.

Lett Appl Microbiol, 2004, 39(1), 13 - 8
Kinetics of enhanced ethanol productivity using raw starch hydrolyzing glucoamylase from Aspergillus niger mutant produced in solid state fermentation; Rajoka MI et al.; AIMS: The present investigation deals with the effect of raw starch hydrolyzing glucoamylase by a derepressed mutant of Aspergillus niger on enhanced productivity of ethanol from uncooked starch under non-aseptic conditions . METHODS AND RESULTS: The parental culture of Aspergillus niger was improved using gamma-ray treatment . One derepressed mutant was isolated after extensive screening and optimization and grown on corn cobs, maize starch, soluble starch and wheat bran solid media moistened with Vogel's salts solution and corn steep liquor . The mutant was 2.5-fold improved over its parent with respect to enzyme productivity, product yield and specific activity . The enzyme from mutated culture was also improved for enzyme properties and could effectively hydrolyze raw starch without the aid of alpha-amylase . Starch hydrolyzed with mutant-derived glucoamylase supported higher volumetric and product yields of ethanol than those of parental and other strains . SIGNIFICANCE AND IMPACT OF THE STUDY: The results of the present study are of commercial value . Ethanol product yield coefficient, and volumetric productivity revealed the hyper-productivity of ethanol from raw starch hydrolyzate obtained with mutant-derived glucoamylase without addition of liquefying alpha-amylase under non-aseptic conditions.

Annu Rev Nutr, 2004, 24, 299 - 326
Vitamin B12 deficiency as a worldwide problem; Stabler SP et al.; Pernicious anemia is a common cause of megaloblastic anemia throughout the world and especially in persons of European or African descent . Dietary deficiency of vitamin B12 due to vegetarianism is increasing and causes hyperhomocysteinemia . The breast-fed infant of a vitamin B12-deficient mother is at risk for severe developmental abnormalities, growth failure, and anemia . Elevated methylmalonic acid and/or total homocysteine are sensitive indicators of vitamin B12-deficient diets and correlate with clinical abnormalities . Dietary vitamin B12 deficiency is a severe problem in the Indian subcontinent, Mexico, Central and South America, and selected areas in Africa . Dietary vitamin B12 deficiency is not prevalent in Asia, except in vegetarians . Areas for research include intermittent vitamin B12 supplement dosing and better measurements of the bioavailability of B12 in fermented vegetarian foods and algae.

Antonie Van Leeuwenhoek, 2000 Apr, 77(3), 281 - 91
Adaptation of methane formation and enzyme contents during growth of Methanobacterium thermoautotrophicum (strain deltaH) in a fed-batch fermentor; Pennings JL et al.; During growth of Methanobacterium thermoautotrophicum in a fed-batch fermentor, the cells are confronted with a steady decrease in the concentration of the hydrogen energy supply . In order to investigate how the organism responds to these changes, cells collected during different growth phases were examined for their methanogenic properties . Cellular levels of the various methanogenic isoenzymes and functionally equivalent enzymes were also determined . Cells were found to maintain the rates of methanogenesis by lowering their affinity for hydrogen: the apparent Km(H2) decreased in going from the exponential to the stationary phase . Simultaneously, the maximal specific methane production rate changed . Levels of H2-dependent methenyl-tetrahydromethanopterin dehydrogenase (H2-MDH) and methyl coenzyme M reductase isoenzyme II (MCR II) decreased upon entry of the stationary phase . Cells grown under conditions that favored MCR II expression had higher levels of MCR II and H2-MDH, whereas in cells grown under conditions favoring MCR I, levels of MCR II were much lower and the cells had an increased affinity for hydrogen throughout the growth cycle . The use of thiosulfate as a medium reductant was found to have a negative effect on levels of MCR II and H2-MDH . From these results it was concluded that M . thermoautotrophicum responds to variations in hydrogen availability and other environmental conditions (pH, growth temperature, medium reductant) by altering its physiology . The adaptation includes, among others, the differential expression of the MDH and MCR isoenzymes.

Antonie Van Leeuwenhoek, 2000 Apr, 77(3), 223 - 8
Saccharomyces bulderi sp . nov., a yeast that ferments gluconolactone; Middelhoven WJ et al.; An unknown yeast species was isolated from maize silage and was determined to be novel on the basis of morphological and physiological characteristics, nucleotide sequence of domain D1/D2 of LSU rDNA and from its electrophoretic karyotype . The name for the proposed new species is Saccharomyces bulderi Middelhoven, Kurtzman et Vaughan-Martini (type strain CBS 8638, NRRL Y-27203, DBVPG 7127) . S . bulderi is closely related to S . barnettii and S . exiguus from which it can be distinguished by having a double vitamin requirement of biotin and thiamine and by no or slow aerobic growth on raffinose, a sugar that on the contrary is fermented rapidly . Gluconolactone is rapidly fermented with ethanol, glycerol and carbon dioxide being the main products.

Klin Lab Diagn, 2004 Apr, (4), 12 - 5
{Clinical, biochemical and ultrasonic changes in the liver in small intestine enzymopathy in children}; Uvarova IO et al.; We followed 70 patients, aged 1 to 11 years: 30 children had primary celiac enzymopathy (CE), the other had secondary fermentopathy represented by the celiac syndrome in 20 subjects and by the disaccharidase insufficiency syndrome (DIS) in another 30 subjects . Distinct disorders were registered in the liver and hepatobiliary system in children with enzymopathy variations of the small intestine (ESI) . Despite a general identity of the clinical signs observed in ESI, the ultrasound signs of the fat hepatosis were more often the case in the celiac disease rather than in the celiac syndrome . The results should be considered in the rational diet- and drug therapy in children with the above pathology.

Curr Biol, 2004 May 25, 14(10), R392 - 4
Evolutionary genomics: yeasts accelerate beyond BLAST; Wolfe K; Two new genome sequences confirm that a whole genome duplication occurred in an ancestor of Saccharomyces cerevisiae . This left a legacy of about 500 pairs of duplicated genes, many of which contribute to this yeast's ability to ferment glucose anaerobically; a few have been evolving so quickly they retain almost no sequence similarity to each other.

Mol Microbiol, 2004 Jun, 52(6), 1703 - 20
The Mycoplasma fermentans prophage phiMFV1: genome organization, mobility and variable expression of an encoded surface protein; Roske K et al.; The approximately 16 kb genome of the Mycoplasma fermentans phiMFV1 prophage is described, and its mobility, replication and effect on the mycoplasma surface phenotype are demonstrated . In various M . fermentans strains, phiMFV1 was either absent or integrated at diverse (and sometimes multiple) chromosomal sites, each marked by a conserved TTTTTA target sequence that is duplicated upon integration . Precise excision, replication of an extrachromosomal form and loss of phiMFV1 from the mycoplasmal genome were documented in a series of clonal derivatives of M . fermentans propagated in culture . Of 18 open reading frames (ORFs) encoded by phiMFV1, most can be ascribed functions related to phage biology, whereas one encodes a unique coiled-coil membrane surface protein, Mem, that was confirmed to be expressed in propagating populations of M . fermentans . With the exception of Mem and other minor ORFs, the striking similarity between the deduced proteomes of phiMFV1 and the recently described phiMAV1 of arthritogenic strains of Mycoplasma arthritidis, along with the prominent gene synteny between these elements, provides the taxonomic basis for a new family of prophage . Their coding features are consistent with long-term residence in mycoplasma genomes and the divergence of species within a phylogenetic clade of mycoplasmas . The unique Mem protein expressed from phiMFV1 and the unique hypothetical surface lipoproteins encoded by phiMAV1 and phiMFV1 also suggest that prophage-associated genes may provide specific, selectable phenotypic traits during co-evolution of mycoplasma species with their respective mammalian hosts . Retention of these labile prophage elements in organisms with such drastically reduced genome sizes implies a significant role in adaptation and survival.

J Agric Food Chem, 2004 Jun 16, 52(12), 3807 - 13
Effect of storage temperature on the stability of anthocyanins of a fermented black carrot (Daucus carota var . L.) beverage: shalgam; Turker N et al.; The effect of temperature on the stability of shalgam anthocyanins stored at 4, 25, and 40 degrees C for 90 days was investigated . The effect of pasteurization and sorbate addition on the anthocyanin stability as compared to control was also studied . The monomeric anthocyanin content and color density decreased with increasing time as a function of storage temperature whereas the percent polymeric color and browning increased . The same trends were observed in control, pasteurized, and sorbate-added shalgam samples . Shalgam anthocyanins consisted of two nonacylated and three acylated cyanidin derivatives . Acylated anthocyanins were more stable when compared to nonacylated ones at all storage temperatures . The activation energies, 11.11-11.64 kcal/mol, were calculated from the reaction rate constants evaluated taking first-order reaction kinetics . The highest anthocyanin retention was observed at 4 degrees C storage temperature with a half-life between 231 and 239 days.

J Agric Food Chem, 2004 Jun 16, 52(12), 3726 - 35
Principal component analysis applied to Fourier transform infrared spectroscopy for the design of calibration sets for glycerol prediction models in wine and for the detection and classification of outlier samples; Nieuwoudt HH et al.; Principal component analysis (PCA) was used to identify the main sources of variation in the Fourier transform infrared (FT-IR) spectra of 329 wines of various styles . The FT-IR spectra were gathered using a specialized WineScan instrument . The main sources of variation included the reducing sugar and alcohol content of the samples, as well as the stage of fermentation and the maturation period of the wines . The implications of the variation between the different wine styles for the design of calibration models with accurate predictive abilities were investigated using glycerol calibration in wine as a model system . PCA enabled the identification and interpretation of samples that were poorly predicted by the calibration models, as well as the detection of individual samples in the sample set that had atypical spectra (i.e., outlier samples) . The Soft Independent Modeling of Class Analogy (SIMCA) approach was used to establish a model for the classification of the outlier samples . A glycerol calibration for wine was developed (reducing sugar content < 30 g/L, alcohol > 8% v/v) with satisfactory predictive ability (SEP = 0.40 g/L) . The RPD value (ratio of the standard deviation of the data to the standard error of prediction) was 5.6, indicating that the calibration is suitable for quantification purposes . A calibration for glycerol in special late harvest and noble late harvest wines (RS 31-147 g/L, alcohol > 11.6% v/v) with a prediction error SECV = 0.65 g/L, was also established . This study yielded an analytical strategy that combined the careful design of calibration sets with measures that facilitated the early detection and interpretation of poorly predicted samples and outlier samples in a sample set . The strategy provided a powerful means of quality control, which is necessary for the generation of accurate prediction data and therefore for the successful implementation of FT-IR in the routine analytical laboratory.

Nat Biotechnol, 2004 Jul, 22(7), 848 - 55 Epub 2004 Jun 06.
Para-position derivatives of fungal anthelmintic cyclodepsipeptides engineered with Streptomyces venezuelae antibiotic biosynthetic genes; Yanai K et al.; PF1022A, a cyclooctadepsipeptide possessing strong anthelmintic properties and produced by the filamentous fungus Rosellinia sp . PF1022, consists of four alternating residues of N-methyl-L-leucine and four residues of D-lactate or D-phenyllactate . PF1022A derivatives obtained through modification of their benzene ring at the para-position with nitro or amino groups act as valuable starting materials for the synthesis of compounds with improved anthelmintic activities . Here we describe the production of such derivatives by fermentation through metabolic engineering of the PF1022A biosynthetic pathway in Rosellinia sp . PF1022 . Three genes cloned from Streptomyces venezuelae, and required for the biosynthesis of p-aminophenylpyruvate from chorismate in the chloramphenicol biosynthetic pathway, were expressed in a chorismate mutase-deficient strain derived from Rosellinia sp . PF1022 . Liquid chromatography-mass spectrometry and NMR analyses confirmed that this approach facilitated the production of PF1022A derivatives specifically modified at the para-position . This fermentation method is environmentally safe and can be used for the industrial scale production of PF1022A derivatives.

Appl Environ Microbiol, 2004 Jun, 70(6), 3521 - 7
Specific 12 beta-hydroxylation of cinobufagin by filamentous fungi; Ye M et al.; Biotransformation of natural products has great potential for producing new drugs and could provide in vitro models of mammalian metabolism . Microbial transformation of the cytotoxic steroid cinobufagin was investigated . Cinobufagin could be specifically hydroxylated at the 12 beta-position by the fungus Alternaria alternata . Six products from a scaled-up fermentation were obtained by silica gel column chromatography and reversed-phase liquid chromatography and were identified as 12 beta-hydroxyl cinobufagin, 12 beta-hydroxyl desacetylcinobufagin, 3-oxo-12 beta-hydroxyl cinobufagin, 3-oxo-12 beta-hydroxyl desacetylcinobufagin, 12-oxo-cinobufagin, and 3-oxo-12 alpha-hydroxyl cinobufagin . The last five products are new compounds . 12 beta-Hydroxylation of cinobufagin by A . alternata is a fast catalytic reaction and was complete within 8 h of growth with the substrate . This reaction was followed by dehydrogenation of the 3-hydroxyl group and then deacetylation at C-16 . Hydroxylation at C-12 beta also was the first step in the metabolism of cinobufagin by a variety of fungal strains . In vitro cytotoxicity assays suggest that 12 beta-hydroxyl cinobufagin and 3-oxo-12 alpha-hydroxyl cinobufagin exhibit somewhat decreased but still significant cytotoxic activities . The 12 beta-hydroxylated bufadienolides produced by microbial transformation are difficult to obtain by chemical synthesis.

Appl Environ Microbiol, 2004 Jun, 70(6), 3467 - 74
Extracellular iron reduction is mediated in part by neutral red and hydrogenase in Escherichia coli; McKinlay JB et al.; Both microbial iron reduction and microbial reduction of anodes in fuel cells can occur by way of soluble electron mediators . To test whether neutral red (NR) mediates iron reduction, as it does anode reduction, by Escherichia coli, ferrous iron levels were monitored in anaerobic cultures grown with amorphous iron oxide . Ferrous iron levels were 19.4 times higher in cultures fermenting pyruvate in the presence of NR than in the absence of NR . NR did not stimulate iron reduction in cultures respiring with nitrate . To explore the mechanism of NR-mediated iron reduction, cell extracts of E . coli were used . Cell extract-NADH-NR mixtures had an enzymatic iron reduction rate almost 15-fold higher than the chemical NR-mediated iron reduction rate observed in controls with no cell extract . Hydrogen was consumed during stationary phase (in which iron reduction was detectable) especially in cultures containing both NR and iron oxide . An E . coli hypE mutant, with no hydrogenase activity, was also impaired in NR-mediated iron reduction activity . NR-mediated iron reduction rates by cell extracts were 1.5 to 2 times higher with hydrogen or formate as the electron source than with NADH . Our findings suggest that hydrogenase donates electrons to NR for extracellular iron reduction . This process appears to be analogous to those of iron reduction by bacteria that use soluble electron mediators (e.g., humic acids and 2,6-anthraquinone disulfonate) and of anode reduction by bacteria using soluble mediators (e.g., NR and thionin) in microbial fuel cells.

Appl Environ Microbiol, 2004 Jun, 70(6), 3392 - 400
Biomass content governs fermentation rate in nitrogen-deficient wine musts; Varela C et al.; Problematic fermentations are common in the wine industry . Assimilable nitrogen deficiency is the most prevalent cause of sluggish fermentations and can reduce fermentation rates significantly . A lack of nitrogen diminishes a yeast's metabolic activity, as well as the biomass yield, although it has not been clear which of these two interdependent factors is more significant in sluggish fermentations . Under winemaking conditions with different initial nitrogen concentrations, metabolic flux analysis was used to isolate the effects . We quantified yeast physiology and identified key metabolic fluxes . We also performed cell concentration experiments to establish how biomass yield affects the fermentation rate . Intracellular analysis showed that trehalose accumulation, which is highly correlated with ethanol production, could be responsible for sustaining cell viability in nitrogen-poor musts independent of the initial assimilable nitrogen content . Other than the higher initial maintenance costs in sluggish fermentations, the main difference between normal and sluggish fermentations was that the metabolic flux distributions in nitrogen-deficient cultures revealed that the specific sugar uptake rate was substantially lower . The results of cell concentration experiments, however, showed that in spite of lower sugar uptake, adding biomass from sluggish cultures not only reduced the time to finish a problematic fermentation but also was less likely to affect the quality of the resulting wine as it did not alter the chemistry of the must.

FEMS Microbiol Lett, 2004 Jun 15, 235(2), 315 - 22
Enhanced expression of aconitase raises acetic acid resistance in Acetobacter aceti; Nakano S et al.; Acetobacter spp . are used for industrial vinegar production because of their high ability to oxidize ethanol to acetic acid and high resistance to acetic acid . Two-dimensional gel electrophoretic analysis of a soluble fraction of Acetobacter aceti revealed the presence of several proteins whose production was enhanced, to various extents, in response to acetic acid in the medium . A protein with an apparent molecular mass of 100 kDa was significantly enhanced in amount by acetic acid and identified to be aconitase by NH2-terminal amino acid sequencing and subsequent gene cloning . Amplification of the aconitase gene by use of a multicopy plasmid in A . aceti enhanced the enzymatic activity and acetic acid resistance . These results showed that aconitase is concerned with acetic acid resistance . Enhancement of the aconitase activity turned out to be practically useful for acetic acid fermentation, because the A . aceti transformant harboring multiple copies of the aconitase gene produced a higher concentration of acetic acid with a reduced growth lag-time.

Bioresour Technol, 2004 Sep, 94(3), 311 - 9
Induction, production, repression, and de-repression of exoglucanase synthesis in Aspergillus niger; Hanif A et al.; The influence of carbon and nitrogen sources on the production of cellulases was investigated . The enzyme production was variable according to the carbon source . Levels of beta-cellobiohydrolase (CBH) were minimal in the presence of even low concentrations of glucose . Enzyme production was stimulated by other carbohydrates . The enzyme is subject to carbon source control by easily metabolizable sugars . Wheat bran and cellulose were the most effective promoters of beta-cellobiohydrolase and filter paperase (FPase) activities respectively, followed by rice bran . Exogenously supplied glucose inhibited the synthesis of the enzyme in cultures of A . niger growing on wheat bran . In defined medium with cellobiose, the cellobiohydrolase titres were 2- to 110-fold higher with cells growing on monomeric sugars and 1.5 times higher than cells growing on other disaccharides . It appeared that synthesis of beta-cellobiohydrolase varied under an induction mechanism, and a repression mechanism which changed the rate of synthesis of beta-cellobiohydrolase and FPase in induced over non-induced cultures . In this organism, substantial synthesis of beta-cellobiohydrolase can be induced by cellobiose, cellodextrin, cellulose or cellulose and hemi-cellulose containing substrates which showed low volumetric substrate uptake rate . The organism required limiting concentration of carbon, nitrogen or phosphorous for production of beta-cellobiohydrolase and FPase . During growth of A . niger on wheat bran, maximum volumetric productivities (Qp) of beta-cellobiohydrolase and FPase were 39.6 and 32.5 IU/lh and were significantly higher than the values reported for some other potent fungi and bacteria . The addition of actinomycin D (a repressor of transcription) and cycloheximide, (a repressor of translation) completely repressed CBH/FPase biosynthesis, suggested that the regulation of CBH synthesis in this organism occurs at both transcriptional and translational level . Thermodynamic studies revealed that the culture exerted protection against thermal inactivation when exposed to different fermentation temperatures.

Bioresour Technol, 2004 Sep, 94(3), 293 - 8
Composition of corn and distillers dried grains with solubles from dry grind ethanol processing; Belyea RL et al.; Increase in the demand for ethanol has resulted in growth in the dry grind (DG) ethanol industry . In DG processing, the whole corn kernel is fermented, resulting in two main coproducts, ethanol and distillers dried grains with solubles (DDGS) . Marketing of DDGS is critical to the economic stability of DG plants . The composition of DDGS can vary considerably; this reduces market value . Factors that cause variation in composition need to be evaluated . The objective was to determine the relationship between composition of corn and composition of DDGS . Samples of corn and DDGS were obtained from a DG ethanol plant and analyzed for protein, fat, starch and other nutrients . Concentrations of protein, fiber and starch were similar to published data for corn but were higher for DDGS . Coefficients of variation for protein fat and fiber concentrations were similar for corn and DDGS . There were no significant correlations between concentrations of components in corn and those in DDGS . Variation in the composition of DDGS was not related to variation in corn composition and probably was due to variation in processing streams or processing techniques . This implies that reducing the variation in composition of DDG will require modification of processing strategies.

Caries Res, 2004 Jul-Aug, 38(4), 341 - 9
Intra-oral acid production associated with eating whole or pulped raw fruits; Beighton D et al.; The hypotheses that raw fruits, whether whole or pulped, were cleared rapidly from the mouth and that the sugars in the whole and pulped fruits are fermented with equal efficiency to acids by the oral microflora were tested in this study . Groups of 7-9 adult subjects chewed 10 g of raw, whole or pulped fruit (apple, banana, orange, pear and pineapple) for 1 min and whole, unstimulated saliva samples were collected during the following 60-min interval . Each saliva sample was assayed for the concentrations of fruit-derived sugars (glucose, fructose and sucrose), fruit-derived acids (malic and citric) and acids which may be produced as a result of bacterial fermentation (acetic, lactic, formic and succinic) . We found the fruit-derived sugars were rapidly cleared from the mouth (within 5 min) . The major bacterially produced acids were lactic and succinic, which reached maximum concentrations in the 5-min sample . There was no significant difference, within a fruit, in the salivary levels of any of the sugars or acids between the raw whole or raw pulped forms . In light of these findings it seems unwise to assume that fruits may be consumed without consideration of their acidogenic potential .

Plant Physiol, 2004 Jun, 135(2), 879 - 90 Epub 2004 Jun 04.
Trehalose mediated growth inhibition of Arabidopsis seedlings is due to trehalose-6-phosphate accumulation; Schluepmann H et al.; Trehalose-6-phosphate (T6P) is required for carbon utilization during Arabidopsis development, and its absence is embryo lethal . Here we show that T6P accumulation inhibits seedling growth . Wild-type seedlings grown on 100 mm trehalose rapidly accumulate T6P and stop growing, but seedlings expressing Escherichia coli trehalose phosphate hydrolase develop normally on such medium . T6P accumulation likely results from much-reduced T6P dephosphorylation when trehalose levels are high . Metabolizable sugars added to trehalose medium rescue T6P inhibition of growth . In addition, Suc feeding leads to a progressive increase in T6P concentrations, suggesting that T6P control over carbon utilization is related to available carbon for growth . Expression analysis of genes from the Arabidopsis trehalose metabolism further supports this: Suc rapidly induces expression of trehalose phosphate synthase homolog AtTPS5 to high levels . In contrast, T6P accumulation after feeding trehalose in the absence of available carbon induces repression of genes encoding T6P synthases and expression of T6P phosphatases . To identify processes controlled by T6P, we clustered expression profile data from seedlings with altered T6P content . T6P levels correlate with expression of a specific set of genes, including the S6 ribosomal kinase ATPK19, independently of carbon status . Interestingly, Suc addition represses 15 of these genes, one of which is AtKIN11, encoding a Sucrose Non Fermenting 1 (SNF1)-related kinase known to play a role in Suc utilization.

Appl Microbiol Biotechnol, 2004 Nov, 65(6), 658 - 63 Epub 2004 Jun 04.
Lactate production in an integrated process configuration: reducing cell adsorption by shielding of adsorbent; Senthuran A et al.; The problem of binding of microbial cells to an adsorbent matrix during in situ recovery of bioproducts from a fermentation broth has been addressed by shielding the adsorbent with a thin layer of a non-ionic polymer . Extractive bioconversion of lactic acid by integrating ion-exchange adsorption with the fermentation stage was studied . The effect of coating of the ion-exchanger with agarose on product recovery and cell adsorption was evaluated . Extractive fermentation with both uncoated and coated resin resulted in an increase in reactor productivity as compared to the normal fermentation . The free cell density in the system with agarose-coated beads was similar to that in control fermentation, but was significantly lower in the system with the uncoated ion-exchanger . Electron microscopic scanning of the bead surface after passage of the fermentation broth showed cells attached to the native adsorbent but not to the coated one.

Carbohydr Res, 2004 Jun 1, 339(8), 1475 - 81
Preparation and structure elucidation of alginate oligosaccharides degraded by alginate lyase from Vibro sp . 510; Zhang Z et al.; Alginate that was purified from the fermentation solution of marine bacteria Vibro sp . 510 under specific reaction conditions was hydrolyzed by alginate lyase . Seven oligosaccharides, including di-, tri- and tetrasaccharides, were isolated through low-pressure, gel-permeation chromatography (LP-GPC) and semipreparative strong-anion exchange (SAX) fast-protein liquid chromatography (FPLC) . The oligosaccharide structures were elucidated based on ESIMS and 2D NMR spectral analysis . The hydrolytic specificity of this alginate lyase to alginate is discussed .

Biotechnol Prog, 2004 May-Jun, 20(3), 818 - 24
Supercritical carbon dioxide extraction of griseofulvin from the solid matrix obtained after solid-state fermentation; Saykhedkar SS et al.; Globally there is an increasing concern to minimize the use of organic solvents, particularly the chlorinated ones because of their suspected human carcinogenicity . The use of ecofriendly carbon dioxide as an alternative to organic solvents would be appropriate in the perspective of green technology . Supercritical carbon dioxide (SC-CO(2)) extraction is suitable for extraction of nonpolar compound with molecular weights less than 400 . Griseofulvin is an antifungal antibiotic having a molecular weight of 353, making it amenable to SC-CO(2) extraction . This work brings out the potential of supercritical carbon dioxide extraction (SCFE) for downstream processing of griseofulvin from the solid matrix obtained after solid-state fermentation (SSF) . The optimized conditions for SCFE of griseofulvin from dried media after SSF were a flow rate of 0.4 L/min, temperature of 60 degrees C, and contact time of 90 min (30 min static + 60 min dynamic) at a pressure of 450-455 bar.

Biotechnol Prog, 2004 May-Jun, 20(3), 764 - 70
Recovery of ammonium lactate and removal of hardness from fermentation broth by nanofiltration; Kang SH et al.; Nanofiltration (NF) was investigated as an alternative to desalting electrodialysis (ED) and ion exchange for the recovery of ammonium lactate from fermentation broth . Three commercial NF membranes, NF45, NF70, and NTR-729HF, were characterized with 50 mM NaCl, MgSO(4), and glucose solutions . NF45 membrane was selected because it showed the lowest rejection of monovalent ion, the highest rejection of divalent ion, and the highest rejection of nonpolar molecule . Effects of the operating pressure were investigated in a range of 100-400 psig, on the flux, lactate recovery, and glucose and magnesium removal from a real fermentation broth containing about 1.0 M of ammonium lactate . The flux and recovery rate increased linearly with the pressure . However, lactate rejection also increased with the pressure, lowering the recovery yield . More magnesium ions and glucose were rejected as the pressure was increased, and at 400 psig, for example, magnesium ion was almost completely rejected, highlighting the chance of obviating the necessity of ion exchange to remove hardness, by using NF instead of desalting ED . Membrane fouling was not so severe as expected, considering the complex nature and a rather high concentration of the fermentation broth treated.

Biotechnol Prog, 2004 May-Jun, 20(3), 744 - 9
Effect of washing on yield in one- and two-step steam pretreatment of softwood for production of ethanol; Soderstrom J et al.; Two-step steam pretreatment of softwood on laboratory scale has previously been shown to result in higher yields than one-step steam pretreatment . In this study, these results are verified on a larger scale . In an industrial process filtration and washing of the material between the two pretreatment steps are difficult without release of pressure . A worst case without filtration or washing was thus investigated to determine the influence of poor washing on the yield of sugars and the formation of byproducts . Steam pretreatment with SO(2) impregnation was investigated using three different procedures . One-step steam pretreatment was performed at 215 degrees C for 5 min . Two different kinds of two-step steam pretreatment were performed at 190 degrees C for 2 min in the first step and at 210 degrees C for 5 min in the second step . In one case the slurry obtained after the first pretreatment step was separated into a liquid and a solid phase, where the water-insoluble solid material was washed with water and then used for pretreatment in the second step . In the other case of two-step steam pretreatment, neither separation nor washing was performed . The pretreated material was evaluated using both enzymatic hydrolysis and fed-batch simultaneous saccharification and fermentation . Both two-step steam pretreatment process configurations investigated resulted in higher yields of ethanol (300 L/ton) than one-step steam pretreatment (227 L/ton) . Separation and washing of the material between the pretreatment steps in the two-step steam pretreatment process did not improve the overall sugar yield, although the formation of sugar degradation products was reduced.

Biotechnol Prog, 2004 May-Jun, 20(3), 737 - 43
Production of phytase (myo-inositolhexakisphosphate phosphohydrolase) by Aspergillus niger van Teighem in laboratory-scale fermenter; Vats P et al.; The growth and production pattern of phytase by a filamentous fungus, Aspergillus niger van Teighem, were studied in submerged culture at varying agitation rates and controlled and uncontrolled pH conditions . Allowing the initial culture to grow under neutral condition with subsequent decline in pH resulted in increased phytase productivity . A maximum of 141 nkat/mL phytase was obtained when the broth pH was maintained at pH 2.5 as compared to 17 nkat/mL units at controlled pH 5.5 . The culture morphology and rheological properties of the fermentation broth significantly varied with the agitation rate . The volumetric oxygen transfer coefficient was determined at different phases of fungal growth during batch fermentation using static gassing out and dynamic gassing out methods . The oxygen transfer coefficient (k(L)a) of the fermenter was found to be 125 h(-)(1) at 500 rpm as compared to 38 h(-)(1) at 200 rpm . The oxygen transfer rates at different phases of growth were significantly affected by cell mass concentration and fungal morphology . During the course of fermentation there was a gradual decline of k(L)a from 97 h(-)(1) on day 2 to 63 h(-)(1) on day 6 of fermentation, after which no significant change was observed . The degree of agitation considerably influenced the culture morphology where shear thinning of filamentous fungus was observed with the increase in agitation.

J Nutr, 2004 Jun, 134(6), 1481 - 6
Dietary fiber stabilizes blood glucose and insulin levels and reduces physical activity in sows (Sus scrofa); de Leeuw JA et al.; The aim of this study was to test whether a diet with a high level of fermentable dietary fiber can stabilize interprandial blood glucose and insulin levels, prevent declines below basal levels, and reduce physical activity in limited-fed breeding sows . Stable levels of glucose and insulin may prevent interprandial feelings of hunger and, consequently, increased activity . Catheterized sows (n = 10) were fed twice daily (0700 and 1900 h) 900 g of a diet with either a low (L-sows) or a high level of fermentable dietary fiber (H-sows; sugarbeet pulp) . Blood samples, taken between feeding times, were analyzed for glucose and insulin levels (basal and area under the curve) and stability of levels (variance and sum of absolute differences between levels in consecutive samples) . The main focus was on samples taken after the postprandial peak . Behavior was videotaped for analysis of postures and posture changes . Basal glucose and insulin levels did not differ between treatments . H-sows had more stable levels than L-sows . Interprandial levels of H-sows were higher than or equal to basal levels . L-sows showed a decline in glucose below basal levels at 1400 h (P < 0.05) . Before 1400 h, no difference in the frequency of posture changes was observed between treatments . After 1400 h, the frequency of posture changes increased more in L-sows than in H-sows . We concluded that sugarbeet pulp as a source of fermentable dietary fiber stabilizes glucose and insulin levels and reduces physical activity in limited-fed sows several hours after feeding . This may indicate a prolonged feeling of satiety.

Sci Total Environ, 2004 Jul 5, 327(1-3), 185 - 96
Occurrence and origin of phosphine in landfill gas; Roels J et al.; A landfill (Hooge Maey, Flanders, Belgium) was subjected to an in-depth study in order to explain the origin of phosphine detected in high amounts in landfill gas, in comparison with biogas from other sources, during a previous study . The spatial and temporal variability of the phosphine concentration in landfill gas was assessed . Twenty-four wells were monitored and differences in phosphine concentration up to one log unit were observed (3.2-32.4 microg/m(3)) . The phosphine concentration in each well was constant in time over a period of 4 months . No correlation was found between the phosphine concentration and methane, carbon dioxide, hydrogen sulfide, ethene or ethane concentration . In a series of laboratory tests, it was shown that phosphine was emitted during batch fermentation tests inoculated with landfill leachate when Fe(0) or Al(0) specimens were added . Conditions favouring corrosion of iron gave rise to higher emissions of phosphine . The phosphine concentration in the headspace of a batch test rose to 1.43 mg/m(3) after 27 days of incubation . Weight loss of corroding steel coupons correlated with phosphine emission . Calculations showed that all phosphine emitted from the 0.005 km(3) landfill (160 g/year) could be attributed to corrosion of metals . No evidence of de novo synthesis could be established.

Int J Food Microbiol, 2004 Jul 1, 94(1), 97 - 103
Lactic acid tolerance determined by measurement of intracellular pH of single cells of Candida krusei and Saccharomyces cerevisiae isolated from fermented maize dough; Halm M et al.; Strains of Candida krusei and Saccharomyces cerevisiae were grown together at 30 degrees C in MYGP broth, pH 2.5, in the presence of 106.4 mM undissociated lactic acid . The two C . krusei strains investigated grew within 48 h from initial counts of 2 x 10(4) to approximately 10(7) cells/ml whereas the two S . cerevisiae strains investigated survived but did not grow in the presence of 106.4 mM undissociated lactic acid at pH 2.5 . To explain the differences in lactic acid tolerance of the two yeast species, we used fluorescence-ratio-imaging microscopy and a perfusion system to determine the short-term intracellular pH (pH(i)) changes in single cells of C . krusei and S . cerevisiae . The changes were investigated both in the presence of low (20.7 mM) and high (106.4 mM) concentrations of undissociated lactic acid . For both the investigated species 20.7 mM undissociated lactic acid did not seem to influence the initial pH(i) which for C . krusei was found to be approximately 8.0 and for S . cerevisiae 6.9-7.5 . For both C . krusei strains, perfusion with 106.4 mM undissociated lactic acid induced only weak short-term pH(i) responses with a decrease in pH(i) of less than one pH unit . Contrary, for both strains of S . cerevisiae perfusion with 106.4 mM undissociated lactic acid resulted in a significant decrease in pH(i) from initially 6.9-7.5 to 6.2-6.4 after 1 min and further to a pH(i) of < or = 5.5 after 3 min after which it remained constant . The results obtained show that C . krusei is more resistant to short-term pH(i) changes caused by lactic acid than S . cerevisiae, and this, in turn, may be part of the explanation why C . krusei is more tolerant to lactic acid than S . cerevisiae.

Dtsch Tierarztl Wochenschr, 2004 Apr, 111(4), 135 - 41
{Influences on fermentation and thiamine metabolism in bovine rumen fluid (in vitro)--part 3: Effects of artificially contaminated hay with Cladosporium herbarum and Fusarium graminearum, respectively}; Holtershinken M et al.; The influence of contaminated hay {Cladosporium herbarum (CL) and Fusarium graminearum (FU), respectively} on fermentation and thiamine metabolism of bovine rumen content was investigated using the longterm rumen simulation technique (RUSITEC) . Six investigation periods 25 days long each were carried out . A nine days feeding period with normal hay was followed by the testphase I (five days) with a mixture of normal and mouldy hay and testphase II (five days) with additive an 0.3 mg thiamine per reaction vessel . The last four days served as regeneration time with normal hay . The following marginal effects of mouldy hay on rumen fermentation patterns could be noted . A) During testphase I: cellulase activity: +10.0% (FU); alternation of the thiamine derivate pattern, but no effect on total thiamine content (CL, FU) . B) During testphase II the results were more obvious: bacterial protein synthesis: -22.6% (CL), -24.4% (FU) . Alternation of the fatty acid pattern: propionate (-7.30% FU), n-butyrate (-3.90% CL, +3.49% FU), n-valerate (-8.5% CH, FU) . Cellulase activity: -17.0% (CL, FU) . But no effect on total thiamine (CL, FU); alternation of the thiamine derivate pattern: more non phosphorylate thiamine . The noted effects on rumen fermentation and thiamine metabolism were not severe enough to be responsible for the development of a CCN.

Biosci Biotechnol Biochem, 2004 May, 68(5), 1051 - 8
Isoflavones found in Korean soybean paste as 3-hydroxy-3-methylglutaryl Coenzyme A reductase inhibitors; Sung JH et al.; 3-Hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase is the rate-limiting enzyme in the biosynthesis of cholesterol in mammals . Some microbial metabolites have been found to be HMG-CoA reductase inhibitors . Korean soybean paste is a unique food fermented by many microorganisms . The enzymatic method using the catalytic domain of Syrian hamster HMG-CoA reductase was employed for the screening of HMG-CoA reductase inhibitors . Soybean paste extract was fractionated by vacuum liquid chromatography . Fractions showing relatively high HMG-CoA reductase inhibition were further purified through Sephadex LH-20 column chromatography and C18 preparative HPLC, and the inhibitory compounds were identified as genistein, daidzein, and glycitein.

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