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 retai