Lett Appl Microbiol, 2005, 40(1), 19 - 23 Contaminant yeast detection in industrial ethanol fermentation must by rDNA-PCR; de Souza Liberal AT et al.; Abstract a.t . de souza liberal, e.a . da silva filho, j.o.f . de morais, d.a . simoes and m.a . de morais jr . 2004.Aims: The present work focuses on the possibility to use conserved primers that amplify yeast ITS1-5.8S-ITS2 ribosomal DNA locus (rDNA) to detect the presence of non-Saccharomyces cerevisiae yeast in fermentation must of bioethanol fermentation process . Methods and Results: Total DNA was extracted from pure or mixed yeast cultures containing different cell concentrations and different contaminant/fermenting yeast concentrations and submitted to PCR . Upon improvement of detection limits and DNA extraction protocol, must samples of distillery were checked for the presence of contaminant yeast . Contaminant rDNA bands were detected only in industrial samples during contamination episodes, but not in noncontaminated must . Conclusions: The method described here could detect the presence of contaminant yeast from industrial must in eight hours after sampling . Significance and Impact of the Study: The improved procedure may help to avoid severe contamination episodes at fermentation industries by decreasing the detection time from 5 days to 8 h and possible quantification of contaminant yeasts that can impose economical loss to the process. J Agric Food Chem, 2004 Dec 29, 52(26), 8268 - 71 Polycyclic aromatic hydrocarbons: pollution and source analysis of a black tea; Lin D et al.; Investigations into the manufacturing process of one kind of black tea revealed that it included five steps: withering, rolling, fermentation, drying, and drying and sorting . A total of 16 polycyclic aromatic hydrocarbons (PAHs) were simultaneously measured in fresh leaves, withered leaves, rolled leaves, fermented leaves, crude black tea, and black tea sampled after each manufacturing stage and in the indoor and outdoor air of the drying house . It was observed that the total contents of the 16 PAHs (SigmaPAHs) in the crude black tea and the black tea were obviously higher than those in the tea leaves sampled after each manufacturing step before the drying stage; the air SigmaPAHs in the drying house were about 100 times higher than those outside the drying house . It can be concluded that quantities of PAHs were released into the drying house from the combustion of pine firewood during the drying stage, and then were absorbed by the tea leaves, thus resulting in the high PAH contents in the black tea. J Agric Food Chem, 2004 Dec 29, 52(26), 8169 - 76 Effects of pu-erh tea on oxidative damage and nitric oxide scavenging; Duh PD et al.; The effects of pu-erh tea, which is prepared by fermentation of tea, on oxidative damage and nitric oxide scavenging, compared with various other brands of tea were investigated . The total antioxidant activity was determined using the Trolox equivalent antioxidant capacity (TEAC) assay . The results showed that TEAC values of the 200 microg/mL water extracts of pu-erh tea (WEPT), green tea, oolong tea, and black tea were 86.3, 85.3, 87.4, and 80.3 (microg/mL), respectively, indicating that WEPT showed a significant antioxidant activity . WEPT, like green tea extract, oolong tea extract, and black tea extract, exhibited a remarkable protective effect in lipid (liposome) and nonlipid (protein and deoxyribose) model systems, implying that it is an inhibitor of lipid and nonlipid oxidative damage . It also exhibited metal-binding ability, reducing power, and scavenging effect for free radicals . Moreover, WEPT showed a decreasing effect on nitric oxide production of lipopolysaccharide-induced RAW 264.7 macrophages . In addition, the results revealed that epicatechin (EC), flavonoid, ascorbic acid, and polyphenolic compounds are present in WEPT, which may partially account for the protective effect on oxidative damage . Thus, WEPT may have potential as an antioxidant and as a nitric oxide scavenging agent. J Agric Food Chem, 2004 Dec 29, 52(26), 8053 - 6 Behavior of a fenhexamid photoproduct during the alcoholic fermentation of Saccharomyces cerevisiae; Cabras P et al.; The fungicide fenhexamid {N-(2,3-dichloro-4-hydroxyphenyl)-1-methylcyclohexanecarboxamide} degraded rapidly by UV or sunlight irradiation, yielding 7-chloro-6-hydroxy-2-(1-methylcyclohexyl)-1,3-benzoxazole (CHB) as a main photoproduct . CHB was isolated, and its effect on alcoholic fermentation of Saccharomyces cerevisiae was studied . The results indicate that the presence of CHB does not affect the extent of alcohol production . After 12 days, the amount of CHB in the fermentation medium decreased by ca . 65% . Only 25% of the missing CHB was recovered unchanged from yeasts, most likely because it was adsorbed on the yeast wall cell . The remaining part degraded during the fermentation process . Glucan and chitin, two potential adsorbents, which constitute yeast cell walls, exhibited affinity for CHB. J Agric Food Chem, 2004 Dec 29, 52(26), 7938 - 43 Glucosinolate derivatives in stored fermented cabbage; Ciska E et al.; The research focused on the glucosinolate (GLS) breakdown products formed during the fermentation of cabbage . A relationship between the contents of degradation products in fermented cabbage and native GLS in raw cabbage was investigated . The effect of fermented cabbage storage on the contents of individual compounds was also assayed . Ascorbigen formed from one of the degradation products of glucobrassicin (indole GLS) was found to be a dominating compound in fermented cabbage . Irrespective of the time of fermented cabbage storage, the content of ascorbigen reached approximately 14 micromol/100 g . Neither the content of isothiocyanates, the major degradation products of aliphatic GLS, nor that of cyanides exceeded 2.5 microM . Storage of cabbage caused periodical increases and decreases in the contents of cyanides and consequent declines in the contents of isothiocyanates . The highest relative contents (expressed as a percentage of the native GLS content) of degradation products--ranging from >70 to 96%--were reported for the products of glucoraphanin degradation, whereas the lowest-- <5% --were reported for the products of sinigrin degradation. Chin J Dig Dis, 2004, 5(3), 115 - 7 Effect of short-chain fatty acids on the proliferation and differentiation of the human colonic adenocarcinoma cell line Caco-2; Fu H et al.; OBJECTIVE: Short-chain fatty acids (SCFA) in the colon may maintain colonocyte differentiation and oppose carcinogenesis . The purpose of this study was to investigate the effects of three SCFA, butyrate, propionate and acetate, on the differentiation, proliferation, and matrix interactions of the Caco-2 human colonic adenocarcinoma cell line . METHODS: Differentiation was assessed by brush border enzyme expression and the doubling time (proliferation) was calculated directly from serial cell counts and by the logarithmic transformation method . Cell motility (migration) was quantitated by the expansion of a confluent Caco-2 monolayer (after release from a constraining fence) across bacteriologic plastic dishes precoated with saturating concentrations of type I collagen . Results were expressed as mean +/- SE and were analyzed by ANOVA and Bonferroni's modified t-test . RESULTS: All three SCFA studied altered the Caco-2 phenotype . Treatment with 10 mmol SCFA significantly prolonged the cell doubling time, promoted brush border enzyme expression (cathepsin C), and inhibited the motility of the Caco-2 cells . CONCLUSIONS: Butyrate, propionate and acetate inhibited the proliferation and motility of a well-differentiated human colonic cancer cell line while promoting the expression of the differentiation marker, cathepsin C . Thus the SCFA produced by bacterial fermentation of dietary fiber may exert a protective effect against the development of colon cancer. J Biol Chem . 2004 Dec 15; {Epub ahead of print} The phosphatidylglycerol/cardiolipin biosynthetic pathway is required for the activation of Inositol phosphosphingolipid phospholipase C, Isc1p, during growth of S . cerevisiae; Vaena de Avalos S et al.; Inositolsphingolipid phospholipase C (Isc1p) is the S . cerevisiae member of the extended family of neutral sphingomyelinases that regulate the generation of bioactive ceramides . Recently, we reported that Isc1p is post-translationally activated in the post-diauxic phase of growth and that it localizes to mitochondria (Vaena de Avalos, Okamoto, and Hannun (2004) JBiolChem 279, 11537-11545) . In this study, the in vivo mechanisms of activation and function of Isc1p were investigated . Deletion of ISC1 resulted in markedly lower growth in non-fermentable carbon sources . Interestingly, the growth defect of isc1D strains resembled that of pgs1D strains, lacking the committed step in the synthesis of phosphatidylglycerol (PG) and cardiolipin (CL), which were shown to activate Isc1p in vitro . Therefore, the role of Pgs1p in activation of Isc1p in vivo was investigated . The results showed that in the pgs1D strain, the growth-dependent activation of Isc1p was impaired, as was the ISC1-dependent increase in the levels of phytoceramide during the post-diauxic phase, demonstrating that the activation of Isc1p in vivo is dependent on PGS1 and/on the mitochondrial phospholipids PG/CL . Mechanistically, loss of Isc1p resulted in lower levels of mitochondrial cytochrome c oxidase subunits cox3p and cox4p, previously established targets of both PG and CL (Ostrander, Zhang, Mileykovskaya, Rho, and Dowhan (2001) JBiolChem 276, 25262-25272), thus suggesting that Isc1p mediates at least some functions downstream of PG/CL . This study provides the first evidence for the mechanism of in vivo activation and function of Isc1p . A model with endogenous PG/CL as the in vivo activator of Isc1p is proposed. Biotechnol Adv, 2005 Jan, 23(1), 41 - 61 Methionine production by fermentation; Kumar D et al.; Fermentation processes have been developed for producing most of the essential amino acids . Methionine is one exception . Although microbial production of methionine has been attempted, no commercial bioproduction exists . Here, we discuss the prospects of producing methionine by fermentation . A detailed account is given of methionine biosynthesis and its regulation in some potential producer microorganisms . Problems associated with isolation of methionine overproducing strains are discussed . Approaches to selecting microorganism having relaxed and complex regulatory control mechanisms for methionine biosynthesis are examined . The importance of fermentation media composition and culture conditions for methionine production is assessed and methods for recovering methionine from fermentation broth are considered. J Appl Microbiol, 2005, 98(1), 114 - 20 Production of H by sulphur-deprived cells of the unicellular cyanobacteria Gloeocapsa alpicola and Synechocystis sp . PCC 6803 during dark incubation with methane or at various extracellular pH; Antal TK et al.; Abstract t.k . antal and p . lindblad . 2004.Aims: To examine sulphur (S) deprivation in combination with the presence of methane (CH(4)) and changes in extracellular pH as a method to enhance in situ hydrogen (H(2)) generation during fermentation in the unicellular non-diazotrophic cyanobacteria Gloeocapsa alpicola and Synechocystis PCC 6803 . Methods and Results: The level of H(2) production, measured using a gas chromatography, was determined in S-deprived cells of G . alpicola and Synechocystis PCC 6803 during fermentation . Starvation on S enhanced the rate of H(2) production by more than fourfold in both strains . S-deprived cyanobacteria were able to maintain maximum rate of H(2) production during at least 8 h of fermentation representing the entire dark period of a day . Increased H(2) production was observed during dark anoxic incubation with a gas phase of 100% CH(4) (up to four times) at lower pH of the medium (5.0-5.5) . Conclusions: S-deprivation in combination with CH(4), added or maybe produced by another micro-organisms, and changes in the pH of the media can be used to further increase the specific capacity of unicellular non-N(2)-fixing cyanobacteria to produce H(2) during fermentation with the overall aim of applying it for outdoor photobiological H(2) production . Significance and Impact of the Study: S-deprivation with respect to H(2) production is well studied in the green algae Chlamydomonas reinhardtii while its application for H(2) production in cyanobacteria is novel . Similarly, the stimulation of H(2) generation in the presence of CH(4) opens up new possibilities to increase the H(2) production . Natural gas enriched with H(2) seems to be a perspective fuel and may be an intermediate step on the pathway to the exploitation of pure biohydrogen. J Appl Microbiol, 2005, 98(1), 106 - 13 Vacuum packing: a model system for laboratory-scale silage fermentations; Johnson HE et al.; Abstract h.e . johnson, r.j . merry, d.r . davies, d.b . kell, m.k . theodorou and g.w . griffith . 2004.Aims: To determine the utility of vacuum-packed polythene bags as a convenient, flexible and cost-effective alternative to fixed volume glass vessels for lab-scale silage studies . Methods and Results: Using perennial ryegrass or red clover forage, similar fermentations (as assessed by pH measurement) occurred in glass tube and vacuum-packed silos over a 35-day period . As vacuum-packing devices allow modification of initial packing density, the effect of four different settings (initial packing densities of 0.397, 0.435, 0.492 and 0.534 g cm(-3)) on the silage fermentation over 16 days was examined . Significant differences in pH decline and lactate accumulation were observed at different vacuum settings . Gas accumulation was apparent within all bags and changes in bag volume with time was observed to vary according to initial packing density . Conclusions: Vacuum-packed silos do provide a realistic model system for lab-scale silage fermentations . Significance and Impact of the Study: Use of vacuum-packed silos holds potential for lab-scale evaluations of silage fermentations, allowing higher throughput of samples, more consistent packing as well as the possibility of investigating the effects of different initial packing densities and use of different wrapping materials. Prikl Biokhim Mikrobiol, 2004 Nov-Dec, 40(6), 680 - 7 {Theoretical bases of optimization of the modes of postharvest treatment of alkaloid-synthesizing medicinal plants}; Ethanol production from non-starch carbohydrates of wheat bran; Department of Chemical Engineering, Lund University, P.O . Box 124, SE-221 00 Lund, SwedenWheat bran (WB), produced worldwide in large quantities as a by-product of the wheat milling industry, constitutes a significant underutilized source of sugars . This paper describes various methods of hydrolyzing the abundant polysaccharides in bran to yield a sugar feedstock suitable for fermentation into bioethanol . Firstly, the starch in the bran was released using amylolytic enzymes . The fibrous material remaining was further hydrolyzed . Acid hydrolysis, heat pretreatment followed by enzymatic hydrolysis and direct enzymatic hydrolysis were compared in terms of total sugar yield and pentose sugar yield . The maximum total sugar yield was achieved when small amounts of acid were added at the pretreatment step prior to enzymatic hydrolysis . This form of pretreatment released most pentosans and significantly enhanced the hydrolysis of cellulose . The overall sugar yield of this combined hydrolysis method reached 80% of the theoretical and it consisted of 13.5g arabinose, 22.8g xylose and 16.7g glucose per 100g starch-free bran. Bioresour Technol, 2005 May, 96(7), 819 - 29 Pipeline transport and simultaneous saccharification of corn stover; Kumar A et al.; Pipeline transport of corn stover delivered by truck from the field is evaluated against a range of truck transport costs . Corn stover transported by pipeline at 20% solids concentration (wet basis) or higher could directly enter an ethanol fermentation plant, and hence the investment in the pipeline inlet end processing facilities displaces comparable investment in the plant . At 20% solids, pipeline transport of corn stover costs less than trucking at capacities in excess of 1.4Mdrytonnes/yr when compared to a mid range of truck transport cost (excluding any credit for economies of scale achieved in the ethanol fermentation plant from larger scale due to multiple pipelines) . Pipelining of corn stover gives the opportunity to conduct simultaneous transport and saccharification (STS) . If current enzymes are used, this would require elevated temperature . Heating of the slurry for STS, which in a fermentation plant is achieved from waste heat, is a significant cost element (more than 5cents/l of ethanol) if done at the pipeline inlet unless waste heat is available, for example from an electric power plant located adjacent to the pipeline inlet . Heat loss in a 1.26m pipeline carrying 2Mdrytonnes/yr is about 5 degrees C at a distance of 400km in typical prairie clay soils, and would not likely require insulation; smaller pipelines or different soil conditions might require insulation for STS . Saccharification in the pipeline would reduce the need for investment in the fermentation plant, saving about 0.2cents/l of ethanol . Transport of corn stover in multiple pipelines offers the opportunity to develop a large ethanol fermentation plant, avoiding some of the diseconomies of scale that arise from smaller plants whose capacities are limited by issues of truck congestion. Biotechnol Lett, 2004 Nov, 26(21), 1671 - 4 Metabolic transition step from ethanol consumption to sugar/ethanol; Ramon-Portugal F et al.; The metabolic pathway shift between only ethanol consumption to both sugar/ethanol consumption was measured by on-line analysis of respiratory quotient of a Saccharomyces cerevisiae . The experiments were carried out in a fed-batch culture under aerobic conditions . During the transition phase, respiratory quotient (RQ) profile shows that sugar can be metabolized through the fermentative pathway even to values of RQ lower than 1. Biotechnol Lett, 2004 Oct, 26(20), 1581 - 4 Selective extraction of acetic acid from the fermentation broth produced by Mannheimia succiniciproducens; Huh YS et al.; Acetic acid is by-product from fermentation processes for producing succinic acid using Mannheimia succiniciproducens . To obtain pure succinic acid from the final fermentation broth, acetic acid was selectively removed based on the different extractability of succinic acid and acetic acid with pH using tri-n-octylamine (TOA) as extractant . When successive batch extractions were performed using 0.25 mol TOA kg(-1) dissolved in 1-octanol at pH 5, the mol ratio of succinic acid to acetic acid before extraction was 4.9 and the final ratio after the fourth batch was 9.4. Biotechnol Lett, 2004 Oct, 26(19), 1533 - 9 Segregation to non-dividing cells in recombinant Escherichia coli fed-batch fermentation processes; Sundstrom H et al.; In Escherichia coli fermentation processes, a drastic drop in viable cell count as measured by the number of colony forming units per ml (c.f.u . ml(-1)) is often observed . This phenomenon was investigated in a process for the production of the recombinant fusion protein, promegapoietin (PMP) . After induction, the number of c.f.u . ml(-1) dropped to approximately 10% of its maximum though the biomass concentration continued to increase . Flow cytometric analysis of viability and intracellular concentration of PMP showed that almost all cells were alive and contributed to the production . Thus, the drop in the number of c.f.u . ml(-1) probably reflects a loss of cell division capability rather than cell death. Biotechnol Lett, 2004 Sep, 26(18), 1403 - 6 Rhizoctonia solani, an elicitor of 6-pentyl-alpha-pyrone production by Trichoderma harzianum in a two liquid phases, extractive fermentation system; Serrano-Carreon L et al.; 6-pentyl-alpha-pyrone (6PP) production by Trichoderma harzianum, in an extractive fermentation system, was elicitated by Rhizoctonia solani . The extent of 6PP elicitation was related to the state of Rhizoctonia and to the Trichoderma inoculum type . The use of non-viable Rhizoctonia solani mycelium in mycelium-inoculated Trichoderma harzianum culture, yielded the maximal 6PP production (474 mg l(-1)) compared to control cultures (147 mg l(-1)) and decreased the process time from 192 to 96 h. Bioorg Med Chem Lett, 2005 Jan 17, 15(2), 353 - 6 Isoterreulactone A, a novel meroterpenoid with anti-acetylcholinesterase activity produced by Aspergillus terreus; Yoo ID et al.; A new seven-membered lactone type meroterpenoid, isoterreulactone A, was isolated from the solid state fermentation of Aspergillus terreus and its structure was established by various spectral analysis . Isoterreulactone A inhibited acetylcholinesterase with an IC(50) value of 2.5muM while did not inhibit butyrylcholinesterase even at 500muM. Biomed Environ Sci, 2004 Sep, 17(3), 273 - 80 Study of a novel antiosteoporosis screening model targeted on cathepsin K; Yang J et al.; OBJECTIVE: To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs . METHODS: To obtain the purified cathepsin K, we cloned the target fragment from the mRNA of human osteosacoma cell line MG63 and demonstrated its correctness through DNA sequencing . Cathepsin K was expressed in a high amount in E . coli after IPTG induction, and was purified to near homogenetity through resolution and column purification . The specificity of the protein was shown by Western blotting experiment . The biological activity of the components in the fermentation broth was assayed by their inhibitory effects on cathepsin K and its analog papain . RESULTS: With the inhibition of papain activity as a screen index, the fermentation samples of one thousand strains of fungi were tested and 9 strains among them showed strong inhibitory effects . The crude products of the fermentation broth were tested for their specific inhibitory effects on the purified human cathepsin K, the product of fungi 2358 shows the highest specificity against cathepsin K . CONCLUSIONS: The compounds isolated from fungi 2358 show the highest biological activity and are worth further structure elucidation and function characterization. Appl Microbiol Biotechnol, 2005 Jan, 66(4), 341 - 51 Epub 2004 Nov 13. Microbial hyaluronic acid production; Chong BF et al.; Hyaluronic acid (HA) is a commercially valuable medical biopolymer increasingly produced through microbial fermentation . Viscosity limits product yield and the focus of research and development has been on improving the key quality parameters, purity and molecular weight . Traditional strain and process optimisation has yielded significant improvements, but appears to have reached a limit . Metabolic engineering is providing new opportunities and HA produced in a heterologous host is about to enter the market . In order to realise the full potential of metabolic engineering, however, greater understanding of the mechanisms underlying chain termination is required. J Microbiol Immunol Infect, 2004 Dec, 37(6), 313 - 21 Effects of culturing conditions on production of D-hydantoinase from recombinant Escherichia coli; Huang JH et al.; The effects of culturing conditions on D-hydantoinase production by a recombinant Escherichia coli strain were investigated using a controlled fed-batch fermentation system . Glucose concentration and pH of the culture broth were maintained at less than 3.3 g/L and at 7.0, respectively, in a 5 L jar fermentor . The optimal composition of the batch medium was glucose, 0.25%; yeast extract, 0.75%; (NH4)2SO4, 0.25%; KH2PO4, 0.2% . The optimal feeding solution was glucose, 60%; yeast extract, 30%; ammonia water, 9% . Following 25-h cultivation, 0.02 mM isopropyl-beta-D-thiogalactopyranoside was added to induce dht gene expression and the temperature was shifted from 32 to 27 degrees C to avoid inclusion body formation . The plasmid-harboring dht gene was found to be stably maintained in the E . coli . Under optimal conditions, a cell density of about 25 g dry cell weight/L and a high volumetric productivity of 8300 U/L/h could be achieved after 48 h culture at agitation and aeration rates of 1000 revolutions per minute and 1 vvm, respectively. Phytother Res, 2004 Nov, 18(11), 884 - 8 Pharmacological effects of fermented red pepper; Choi YM et al.; The pharmacological effects were investigated of fermented red pepper (HF-S), which consisted of 14.7% carbohydrate, 1.5% lipid, 4.9% protein, 0.3% ash, 78.2% moisture content, with 0.15% capsaicin and 0.06% dihydrocapsaicin . Oral administration of 0.25 mL HF-S for 3 weeks produced significant changes of the perirenal fat pad weight compared with the HF-control group, suggesting a suppressive effect on lipid accumulation and a significant decrease in the risk of arteriosclerosis . The HF-S (0.25 mL) group also showed a lower plasma TG, TC level and atherogenic index than that of the HF-control . In addition, the HF-S (0.25 mL) group showed a marked increase in the production of glutathione, which is the major endogenous antioxidant, and a decrease in the production of lipid peroxide as the product of chemical damage by oxygen free radicals . It is assumed that the effect of HF-S might relate to high glutathione production on the suppression of lipid peroxidation . HF-S stimulated not only the proliferation of macrophages (as high as the positive control, LPS at 1000 microg/mL) but also mitogenic activity (1.2-fold of LPS at 100 microg/mL) . Proc Natl Acad Sci U S A, 2004 Dec 28, 101(52), 18036 - 41 Epub 2004 Dec 13. Genome sequence of the deep-sea gamma-proteobacterium Idiomarina loihiensis reveals amino acid fermentation as a source of carbon and energy; Hou S et al.; We report the complete genome sequence of the deep-sea gamma-proteobacterium, Idiomarina loihiensis, isolated recently from a hydrothermal vent at 1,300-m depth on the Loihi submarine volcano, Hawaii . The I . loihiensis genome comprises a single chromosome of 2,839,318 base pairs, encoding 2,640 proteins, four rRNA operons, and 56 tRNA genes . A comparison of I . loihiensis to the genomes of other gamma-proteobacteria reveals abundance of amino acid transport and degradation enzymes, but a loss of sugar transport systems and certain enzymes of sugar metabolism . This finding suggests that I . loihiensis relies primarily on amino acid catabolism, rather than on sugar fermentation, for carbon and energy . Enzymes for biosynthesis of purines, pyrimidines, the majority of amino acids, and coenzymes are encoded in the genome, but biosynthetic pathways for Leu, Ile, Val, Thr, and Met are incomplete . Auxotrophy for Val and Thr was confirmed by in vivo experiments . The I . loihiensis genome contains a cluster of 32 genes encoding enzymes for exopolysaccharide and capsular polysaccharide synthesis . It also encodes diverse peptidases, a variety of peptide and amino acid uptake systems, and versatile signal transduction machinery . We propose that the source of amino acids for I . loihiensis growth are the proteinaceous particles present in the deep sea hydrothermal vent waters . I . loihiensis would colonize these particles by using the secreted exopolysaccharide, digest these proteins, and metabolize the resulting peptides and amino acids . In summary, the I . loihiensis genome reveals an integrated mechanism of metabolic adaptation to the constantly changing deep-sea hydrothermal ecosystem. Comp Biochem Physiol A Mol Integr Physiol, 2004 Dec, 139(4), 513 - 9 Effects of cellulose, carboxymethylcellulose and inulin fed to rats as single supplements or in combinations on their caecal parameters; Juskiewicz J et al.; We compared the effect of diets containing different nondigestible carbohydrates: cellulose (C), inulin (IN) and carboxymethylcellulose (CMC) as single supplements or in dietary combination on caecal physiology of rats . Sixty male Wistar rats (Rattus norvegicus) were divided into five groups and for 4 weeks were fed a casein diet with the compared carbohydrates (4% of diet) or a combination of IN+C or IN+CMC (both 4+4%) . Diet intake and FCR index remained unaffected by the treatments, whereas IN improved the body weight gain of rats compared to CMC . Compared to C group, all diets containing IN and CMC decreased the caecal pH as well as enlarged the caecum, thus increasing the weights of contents and tissue, especially upon CMC treatment . Rats given carboxymethylcellulose (CMC and IN+CMC groups) had watery caecal digesta, and some of them suffered from diarrhoea . In the case of CMC, the caecal enlargement was due to tissue hypertrophy and digesta accumulation mostly in response to an increased bulk of contents . Unlike C+IN, the dietary combination of CMC- and inulin-enhanced fermentation in the caecum of rats, however the proportion of acetate, propionate and butyrate was less beneficial . Compared to CMC, inulin gave a higher concentration of SCFA, especially of butyrate and propionate . The action of inulin in the caecum of rats could be pronounced by dietary treatment combined with CMC. Comp Biochem Physiol A Mol Integr Physiol, 2004 Dec, 139(4), 449 - 59 Intake, ingesta retention, particle size distribution and digestibility in the hippopotamidae; Clauss M et al.; Although several aspects of the digestive physiology of the hippopotamidae-non-ruminating foregut fermenters-have been described, ingesta kinetics and passage characteristics of these species are not well understood . The most outstanding feature of the hippo digestive physiology reported so far is the very long mean ingesta retention times (MRTs) measured by Foose {Foose, T., 1982 . Trophic strategies of ruminant versus nonruminant ungulates . PhD dissertation, University of Chicago, Chicago.} . Since those data had been investigated with animals without water access, we intended to measure MRT in hippos which were allowed to enter water pools during the night . MRT parameters as well as dry matter (DM) digestibility were determined in four common (Hippopotamus amphibius) and four pygmy hippos (Hexaprotodon liberiensis) on two different diets each using cobalt ethylendiamintetraacetate (Co-EDTA) as a fluid, chromium (Cr)-mordanted fibre (<2 mm) as a particle and acid detergent lignin (ADL) as an internal digestibility marker . Four of the animals additionally received cerium (Ce)-mordanted fibres (2-10 mm) as particle markers . Total MRTs for fluids and particles ranged between 20-35 and 48-106 h in the common and between 13-39 and 32-107 h in the pygmy hippos . The difference between fluid and particle retention was greater than usually reported in ruminants . Excretion patterns of the markers differed from those usually observed in ruminants but resembled those reported for macropods (kangaroos), indicating a plug-flow reactor-like physiology in the hippo forestomach (FRST) . This finding complements other described similarities between the macropod and the hippo forestomach . The measurements of larger particle retention profiles suggest that in the hippo, larger particles might be excreted either faster or at the same rate as smaller particles, indicating a general difference between ruminants and hippos with respect to differential particle retention . The digestive physiology of hippos is characterised by a generally low food intake, long ingesta retention times and dry matter digestibilities lower than reported in ruminants . Moderate digestibilities in spite of long retention times might be the result of the generally high average ingesta particle size in hippos . The comparatively easy management of pygmy hippos, together with the significant correlations between food intake, MRT and digestibility in the pygmy hippos of this study, recommends this species for further studies on the interplay of these parameters in herbivore digestive physiology. Biochemistry, 2004 Dec 21, 43(50), 15884 - 90 Identification of domains within megalomicin and erythromycin polyketide synthase modules responsible for differences in polyketide production levels in Escherichia coli; Murli S et al.; The megalomicin and erythromycin polyketide synthases (PKSs) produce the same aglycon product, 6-deoxyerythronolide B (6-dEB) . Both PKSs were examined in an Escherichia coli strain metabolically engineered to support complex polyketide biosynthesis . Production of 6-dEB in shake flask fermentations was undetectable by mass spectrometry in the strain expressing the megalomicin (Meg) PKS genes, whereas 31 mg/L 6-dEB was produced by the strain with the erythromycin (DEBS) PKS . The genes for each of the three subunits comprising the PKSs were expressed in different combinations from three compatible expression vectors (e.g., DEBS1, DEBS2, and MegA3) to identify two Meg PKS subunits, MegA1 and MegA3, which conferred lower 6-dEB titers than their DEBS counterparts . Comparison of protein expression levels and 6-dEB titers by engineered hybrid DEBS/Meg PKS genes further defined regions within modules 2 and 6 of MegA1 and MegA3, respectively, which limit protein expression and 6-dEB production in E . coli . Meg module 2 + TE (M2 + TE) and a hybrid DEBS M2/Meg M2 + TE protein were engineered and purified for in vitro comparisons with DEBS M2 + TE . The specific activity of the hybrid M2 + TE was approximately 16-fold lower than DEBS M2 + TE and only twice as high as the Meg M2 + TE enzyme in diketide elongation assays . Since the hybrid M2 worked comparably to DEBS M2 in vivo, this suggests that boosting subunit concentration could serve as a useful approach to overcome enzyme deficiencies in heterologous polyketide production. Nat Prod Res, 2004 Dec, 18(6), 529 - 35 Biotransformation of (+)-androst-4-ene-3,17-dione; Choudhary MI et al.; Fermentation of (+)-androst-4-ene-3,17-dione (1) with Curvularia lunata for 10 days yielded five oxidative and reductive metabolites, androsta-1,4-diene-3,17-dione (2), 17beta-hydroxyandrosta-1,4-dien-3-one (3), 11alpha-hydroxyandrost-4-ene-3,17-dione (4), 11alpha,17beta-dihydroxyandrost-4-en-3-one (5) and 15alpha-hydroxyandrosta-1,4-dien-17-one (6) . The structures of these metabolites were elucidated on the basis of spectroscopic techniques . These microbially transformed products were assayed against the clinically important enzymes, tyrosinase and prolyl endopeptidase. Curr Opin Drug Discov Devel, 2004 Nov, 7(6), 869 - 81 Toward a commercial synthesis of (+)-discodermolide; Mickel SJ; In this review some of the criteria required for a viable chemical (+)-discodermolide production process are discussed by critical evaluation of the available literature approaches . A further route involving the use of polyketide synthase to produce (+)-discodermolide fragments by fermentation is also described . Both approaches have advantages and disadvantages and require significant optimization in order to achieve commercialization of this important natural product. Biotechnol Bioeng, 2005 Jan 20, 89(2), 243 - 51 Estimating optimal profiles of genetic alterations using constraint-based models; Gadkar KG et al.; Metabolic engineering involves application of recombinant DNA methods to manipulate metabolic networks to improve cellular properties . It is critical that the genetic alterations be performed in an optimal manner to maximize profit . In addition to the product yield, productivity consideration is also critical, especially for the production of bulk chemicals such as 1,3-propanediol . In this work, we demonstrate that it is suboptimal from the standpoint of productivity to induce genetic alteration at the start of the production process . A bilevel optimization scheme is formulated to determine the optimal temporal flux profile for the manipulated reaction . In the first case study, an optimal flux in the reaction catalyzed by glycerol kinase is determined to maximize the glycerol production at the end of a 6-h batch cultivation of Escherichia coli under aerobic conditions . The final glycerol concentration is 30% higher for the optimal flux profile compared with having an active flux during the entire batch . The effect of the mass transfer coefficient on the optimal profile and the glycerol concentration is also determined . In the second case study, the anaerobic batch fermentation of the ldh(-) strain of Escherichia coli is considered . The optimal flux in the acetate pathway is determined to maximize the final ethanol concentration . The optimal flux results in higher ethanol concentration (11.92 mmol L(-1)) compared to strains with no acetate flux (8.36 mmol L(-1)) and fully active acetate flux (6.22 mmol L(-1)) . We also examine the effects of growth inhibition due to high ethanol concentrations and variations in final batch time on ethanol production . (c) 2004 Wiley Periodicals, Inc. Bioprocess Biosyst Eng . 2004 Dec 9; {Epub ahead of print} Influence of dissolved oxygen and shear conditions on clavulanic acid production by Streptomyces clavuligerus; Rosa JC et al.; Clavulanic acid (CA), a potent beta-lactamase inhibitor, is produced by a filamentous bacterium . Here, the effect of DO and shear, expressed as impeller tip velocity, on CA production was examined . Cultivations were performed in a 4 L fermentor with speeds of 600, 800 and 1,000 rpm and a fixed air flow rate (0.5 vvm) . Also, cultivation with automatic control of dissolved oxygen, at 50% air saturation, by varying stirrer speed and using a mixture of air and O(2) (10% v/v) in the inlet gas, and a cultivation with fixed stirrer speed of 800 rpm and air flow rate of 0.5 vvm, enriched with 10% v/v O(2), were performed . Significant variations in CA titer, CA production rate and O(2) uptake-rate were observed . It was also found that the DO level has no remarkable effect on CA production once a critical level is surpassed . The most significant improvement in CA production was related to high stirrer speeds. Bull Hist Med, 2004 Winter, 78(4), 767 - 803 Motus Tonicus: Georg Ernst Stahl's formulation of tonic motion and early modern medical thought; Chang KM; This paper places in multiple contexts Stahl's formulation of tonic motion, a contractive and relaxative movement of body tissues that was thought to moderate the circulatory blood flowing through their porous structure . The paper analyzes Stahl's theory, elucidates its role in connecting his physiology and pathology, and situates its formulation in his conceptual development as well as the intellectual history of early modern medicine . The theory was at first a post-Harveyan attempt to explain occasional uneven blood flows; it was then expanded to account for the mechanism of blood circulation and metabolism, and formed a fundamental part of Stahl's effort to present a theory of animal heat and fever that would replace the traditional Galenic and fermentational theories . Tonic motion constituted the most important device that counteracted the ineluctable, unceasing corruption of the body, as dictated by its chemical nature; it thus qualified as the preeminent form of what Stahl considered vital motions. Appl Biochem Biotechnol, 2004 Dec, 119(3), 241 - 78 Early phase process scale-up challenges for fungal and filamentous bacterial cultures; Junker BH et al.; Culture pelleting and morphology has a strong influence on process productivity and success for fungal and filamentous bacterial cultures . This impact is particularly evident with early phase secondary metabolite processes with limited process definition . A compilation of factors affecting filamentous or pelleting morphology described in the literature indicates potential leads for developing process-specific control methodologies . An evaluation of the factors mediating citric acid production is one example of an industrially important application of these techniques . For five model fungal and filamentous bacterial processes in an industrial fermentation pilot plant, process development strategies were developed and effectively implemented with the goal of achieving reasonable fermentation titers early in the process development cycle . Examples of approaches included the use of additives to minimize pelleting in inoculum shake flasks, the use of large-volume frozen bagged inoculum obtained from agitated seed fermentors, and variations in production medium composition and fermentor operating conditions . Results were evaluated with respect to productivity of desired secondary metabolites as well as process scalability . On-line measurements were utilized to indirectly evaluate the cultivation impact of changes in medium and process development . Key laboratory to pilot plant scale-up issues also were identified and often addressed in subsequent cultivations. Proc Natl Acad Sci U S A, 2004 Dec 21, 101(51), 17593 - 8 Epub 2004 Dec 08. Fermented beverages of pre- and proto-historic China; McGovern PE et al.; Chemical analyses of ancient organics absorbed into pottery jars from the early Neolithic village of Jiahu in Henan province in China have revealed that a mixed fermented beverage of rice, honey, and fruit (hawthorn fruit and/or grape) was being produced as early as the seventh millennium before Christ (B.C.) . This prehistoric drink paved the way for unique cereal beverages of the proto-historic second millennium B.C., remarkably preserved as liquids inside sealed bronze vessels of the Shang and Western Zhou Dynasties . These findings provide direct evidence for fermented beverages in ancient Chinese culture, which were of considerable social, religious, and medical significance, and help elucidate their earliest descriptions in the Shang Dynasty oracle inscriptions. Bioresour Technol, 2005 Apr, 96(6), 673 - 86 Features of promising technologies for pretreatment of lignocellulosic biomass; Mosier N et al.; Cellulosic plant material represents an as-of-yet untapped source of fermentable sugars for significant industrial use . Many physio-chemical structural and compositional factors hinder the enzymatic digestibility of cellulose present in lignocellulosic biomass . The goal of any pretreatment technology is to alter or remove structural and compositional impediments to hydrolysis in order to improve the rate of enzyme hydrolysis and increase yields of fermentable sugars from cellulose or hemicellulose . These methods cause physical and/or chemical changes in the plant biomass in order to achieve this result . Experimental investigation of physical changes and chemical reactions that occur during pretreatment is required for the development of effective and mechanistic models that can be used for the rational design of pretreatment processes . Furthermore, pretreatment processing conditions must be tailored to the specific chemical and structural composition of the various, and variable, sources of lignocellulosic biomass . This paper reviews process parameters and their fundamental modes of action for promising pretreatment methods. Mycol Res, 2004 Nov, 108(Pt 11), 1291 - 300 Conspecificity of the cerulenin and helvolic acid producing 'Cephalosporium caerulens', and the hypocrealean fungus Sarocladium oryzae; Bills GF et al.; Fermentation processes for the biochemical reagents cerulenin and helvolic acid employ 'Cephalosporium caerulens,' an invalidly published designation that has been used for more than 40 years . However, its identity has never been critically examined because strains were unavailable from major culture collections . An authentic strain of C . caerulens', derived from the original strain KF-140, was recently found and compared to Sarocladium oryzae, another Acremonium-like fungus which also produces cerulenin and helvolic acid . Morphological comparisons, rDNA sequence data, and chromatography of secondary metabolites established that 'C . caerulens' and S . oryzae are conspecific . Sequence data from ribosomal DNA genes indicated S . oryzae belongs to the Hypocreales and is allied with members of the Ceratostomataceae, Scopinella species, Emericellopsis species and certain Acremonium-like anamorphs of uncertain familial relationships . At least two of the isolates of S . oryzae produced titres of cerulenin and helvolic acid similar to those of KF-140 . This finding demonstrates that manufacture of cerulenin need not be limited to the original strain. Int J Oncol, 2005 Jan, 26(1), 233 - 9 Gene expression associated with the decrease in malignant phenotype of human liver cancer cells following stimulation with a histone deacetylase inhibitor; Wakabayashi K et al.; Sodium butyrate is a short-chain fatty acid produced by fermentation in the gastrointestinal tract . It induces differentiation of several kinds of cancer by inhibiting histone deacetylase activity . We have reported that butyrate stimulates hepatocellular carcinoma cells into their normal phenotype . Since sodium butyrate affects both differentiation and apoptosis, we investigated expression of bcl-2-related genes in a human hepatocellular carcinoma cell line HCC-T . The expression of anti-apoptotic Bcl-2 and Mcl-1/EAT was up-regulated 4 h after the treatment, while pro-apoptotic Bax expression did not change . Gene expressions in the early stage of butyrate-stimulation were investigated by the differential display assay and the cDNA expression array . Laminin and keratin 18 were increased 6 h after the stimulation with sodium butyrate . The results of cDNA expression array revealed up-regulation of cell cycle inhibitory genes such as cyclin-dependent kinase 4 inhibitor, and interferon-related genes such as STAT2 and 3, while down-regulation of cyclin-dependent kinase 2 and cyclin E . Up-regulated production of p21WAF-1 and Mcl-1/EAT was also confirmed by Western blotting . The cytoskeletal change indicated by up-regulation of laminin and keratin 18 may be an important factor in the decrease in malignant phenotype of cancer cells . Up-regulation of interferon-related genes indicated that butyrate-treatment might induce a similar phenotypic change to that induced by type 1 interferons . This study suggests several target genes for the future gene therapy of cancer or genes preventing cancer development from pre-malignant tissues. J Biomol Tech, 2004 Dec, 15(4), 317 - 24 Determination of amino acids in cell culture and fermentation broth media using anion-exchange chromatography with integrated pulsed amperometric detection; Hanko VP et al.; Anion-exchange chromatography with integrated pulsed amperometric detection (AE-IPAD) separates and directly detects amino acids, carbohydrates, alditols, and glycols in the same injection without pre- or post-column derivatization . These separations use a combination of NaOH and NaOH/sodium acetate eluents . We previously published the successful use of this technique, also known as AAA-Direct, to determine free amino acids in cell culture and fermentation broth media . We showed that retention of carbohydrates varies with eluent NaOH concentration differently than amino acids, and thus separations can be optimized by varying the initial NaOH concentration and its duration . Unfortunately, some amino acids eluting in the acetate gradient portion of the method were not completely resolved from system-related peaks and from unknown peaks in complex cell culture and fermentation media . In this article, we present changes in method that improve amino acid resolution and system ruggedness . The success of these changes and their compatibility with the separations previously designed for fermentation and cell culture are demonstrated with yeast extract-peptone-dextrose broth, M199, Dulbecco's modified Eagle's (with F-12), L-15 (Leibovitz), and McCoy's 5A cell culture media. J Anim Physiol Anim Nutr (Berl), 2004 Dec, 88(11-12), 401 - 11 Rumen odd and branched chain fatty acids in relation to in vitro rumen volatile fatty acid productions and dietary characteristics of incubated substrates; Vlaeminck B et al.; Summary A first aim of this batch in vitro experiment (21 h) was to use changes in odd and branched chain fatty acid (OBCFA) patterns to suggest shifts in microbial populations, associated with four types of incubated whole dairy cow diets . Principal component analysis suggested higher dietary starch increased the proportion of C(15:0) and C(17:0), whereas increased neutral detergent fibre content was positively related to anteiso C(15:0) concentrations, which is in agreement with the importance of these fatty acids in respectively amylolytic and cellulolytic bacteria . A second aim of the experiment was to relate rumen volatile fatty acid proportions to OBCFA by principal component regression and to compare these relations with predictions based on diet proximate composition . The R(2) values achieved for the regressions between acetate, propionate and butyrate, and OBCFA were 79.6%, 86.6% and 84.9% respectively . Moreover, in the current study, predictions of the rumen fermentation pattern showed higher R(2) (p < 0.01) when based on OBCFA compared with proximate feed composition . If relations persist in vivo, there could be scope for milk OBCFA to predict the supply of specific rumen nutrients. J Anim Physiol Anim Nutr (Berl), 2004 Dec, 88(11-12), 367 - 80 Pectin in pig nutrition, a comparative review; Drochner W et al.; Summary A comprehensive review is given on pectin effects in pigs . Its effects on feed passage and digestion in the small intestine, the composition of ileal digesta and fermentation in the colon are described . Pectins promote fermentation at the terminal ileum and in the first part of the colon . It is shown, that pectin opposite to cellulose is degraded to high extents in the colon . Dietetic effects in relation to composition of ileocaecal digesta, endogenous secretions, buffering and pH-stability, high fermentability of organic matter in the colon and resulting volatile fatty acids (VFA) in the cranial part of the colon and even its possible interactions to colonization of the gut by infectious bacteria are described . Pectin effects concerning volume of the intestinal tract, distribution of VFA and lactic acid, role of endogenous secretions are described . The negative effect of pectins on praecaecal digestibility, their high degradation rates in the large intestine and consequences for their dietetic effects are discussed. Food Chem Toxicol, 2005 Jan, 43(1), 1 - 19 Effect of ethanol on the tumorigenicity of urethane (ethyl carbamate) in B6C3F(1) mice; Beland FA et al.; Urethane is a carcinogen to which there is widespread exposure through the consumption of fermented foods and alcoholic beverages . In this study, we have assessed the carcinogenicity of urethane in combination with ethanol . Male and female B6C3F(1) mice (48 mice per sex per group) were exposed to 0, 10, 30, or 90ppm urethane in the presence of 0%, 2.5%, or 5% ethanol in drinking water ad libitum for two years, at which time the extent of tumorigenesis was assessed . Additional mice (four per sex per group) received the same doses for four weeks to assess serum levels of urethane and ethanol, DNA adduct formation, and the induction of microsomal cytochromes P450, cell proliferation, and apoptosis . Urethane decreased cell replication in the livers of female, but not male, mice, decreased cell replication in the lungs of both sexes, and induced cytochrome P450 2E1 in the livers of female mice . Hepatic levels of the DNA adduct 1,N(6)-ethenodeoxyadenosine were increased by exposure to urethane and decreased by treatment with ethanol . Animal weights and survival were not affected by ethanol; in contrast, urethane administration decreased body weights and survival . Urethane caused dose-dependent increases in liver, lung, and harderian gland adenoma or carcinoma and hemangiosarcoma of the liver and heart in both sexes, mammary gland and ovarian tumors in females, and squamous cell papilloma or carcinoma of the skin and forestomach in males . The increase in hepatocellular tumors occurred in a relatively linear manner and was attributed to the formation of 1,N(6)-ethenodeoxyadenosine in hepatic DNA coupled with an increase in cell replication . Hemangiosarcomas were observed only at the 90ppm urethane dose and were probably a result of high-dose urethane-induced toxicity . Lung alveolar/bronchiolar and harderian gland adenoma or carcinoma increased in a relatively linear manner, suggestive of a genotoxic mechanism for tumor induction . Ethanol induced a dose-dependent trend in hepatocellular adenoma or carcinoma in male mice, with the incidence being marginally increased at the highest dose . In female mice administered 10ppm and 90ppm urethane, ethanol caused dose-related increases in alveolar/bronchiolar adenoma or carcinoma and hemangiosarcoma of the heart, respectively . This may be due to ethanol decreasing the first-pass clearance of urethane, thus, increasing systemic distribution . In male mice a different relationship was observed: ethanol caused a dose-related decrease in alveolar/bronchiolar and harderian gland adenoma or carcinoma in mice administered 30ppm urethane. Water Sci Technol, 2004, 50(9), 245 - 53 Municipal sludge management and disposal in South Korea: status and a new sustainable approach; Ahn YH et al.; Based on figures from 2002, 5216 ton/d of the municipal sludge is produced from 184 large municipal wastewater treatment plants in 111 cities with total treatment capacity of 19,229,745 m3/d . Even though the large amount of sludge disposal has depended greatly on ocean disposal and landfills until recently, the fraction of sludge reuse has gradually increased from 2.7% to 7%, since 1991 . Due to a need of resources recovery from the sludge, high cost requirement of incineration and legislative regulation, recent new research is mainly focused on resources recovery and its reuse from the municipal sludge, such as high performance acid fermenter with pathogen reduction, crystallization (struvite and hydroxyapatite) using waste lime, cofermentation of municipal sludge with food waste, aerobic composting with P crystallization, vermistabilization, lime treatment, etc . Current research and practical activities with some efforts for the new technical development as well as environmental law and regulation are reviewed. Eur J Immunol, 2005 Jan, 35(1), 282 - 9 Toll-like receptor 6-independent signaling by diacylated lipopeptides; Buwitt-Beckmann U et al.; Bacterial lipopeptides are strong immune modulators that activate early host responses after infection as well as initiating adjuvant effects on the adaptive immune system . These lipopeptides induce signaling in cells of the immune system through Toll-like receptor 2 (TLR2)-TLR1 or TLR2-TLR6 heteromers . So far it has been thought that triacylated lipopeptides, such as the synthetic N-palmitoyl-S-{2,3-bis(palmitoyloxy)-(2RS)-propyl}-(R)-cysteine (Pam3)-CSK4, signal through TLR2-TLR1 heteromers, whereas diacylated lipopeptides, like the macrophage-activating lipopeptide from Mycoplasma fermentans (MALP2) or S-{2,3-bis(palmitoyloxy)-(2RS)-propyl}-(R)-cysteine (Pam2)-CGNNDESNISFKEK, induce signaling through TLR2-TLR6 heteromers . Using new synthetic lipopeptide derivatives we addressed the contribution of the lipid and, in particular, the peptide moieties with respect to TLR2 heteromer usage . In contrast to the current model of receptor usage, not only triacylated lipopeptides, but also diacylated lipopeptides like Pam2CSK4 and the elongated MALP2 analog Pam2CGNNDESNISFKEK-SK4 (MALP2-SK4) induced B lymphocyte proliferation and TNF-alpha secretion in macrophages in a TLR6-independent manner as determined with cells from TLR6-deficient mice . Our results indicate that both the lipid and the N-terminal peptides of lipoproteins contribute to the specificity of recognition by TLR2 heteromers and are responsible for the ligand-receptor interaction on host cells. Appl Microbiol Biotechnol . 2004 Dec 3; {Epub ahead of print} Different control mechanisms regulate glucoamylase and protease gene transcription in Aspergillus oryzae in solid-state and submerged fermentation; Te Biesebeke R et al.; Solid-state fermentation (SSF) with Aspergillus oryzae results in high levels of secreted protein . However, control mechanisms of gene expression in SSF have been only poorly studied . In this study we show that both glucoamylase (glaB) and protease (alpA, nptB) genes are highly expressed during surface cultivation on wheat-based solid medium, and even higher during cultivation on wheat kernels . In wheat-based liquid medium, low levels of gene expression are observed . Typical SSF cultivation conditions, such as low water activity and the formation of aerial hyphae, did not contribute to the high-level gene expression on wheat-based solid medium . Analysis of wheat-based solid and liquid cultivations showed differences in carbon and nitrogen utilisation and external pH . The results presented show that the difference in regulation of transcription of the alpA and nptB genes in wheat-based liquid and solid medium could be pH dependent, involving a pH-dependent transcription regulator . The results obtained suggest that the difference in regulation of transcription of the glaB gene in wheat-based liquid and solid medium is caused by a difference in carbohydrate degradation and consumption under the different culture conditions. Bioprocess Biosyst Eng . 2004 Dec 1; {Epub ahead of print} Control of endotoxin release in Escherichia coli fed-batch cultures; Svensson M et al.; High amounts of outer membrane (OM) components were released in glucose-limited fed-batch (GLFB) cultures at 37 degrees C at specific growth rates approaching 0.05 h(-1) . Endotoxin analyses from a 20 degrees C GLFB culture gave similar results . An alternative fermentation technique, the temperature-limited fed-batch (TLFB) technique, reduced the endotoxin concentration in a culture with a biomass concentration of 30 g l(-1) from the 850 mg l(-1) in traditional GLFB cultures to about 20 mg l(-1) . The TLFB technique uses the temperature to regulate the dissolved oxygen tension, while all substrate components are unregulated . It appears to be severe glucose limitation that triggers the extensive release of endotoxins rather than a low growth rate . Furthermore, it is not the low temperature that stabilizes the OM when using the TLFB technique . Simulations and experimental data show that this technique results in the same biomass productivity as the GLFB technique. Biotechnol Prog, 2004 Nov-Dec, 20(6), 1880 - 4 Extractive cultivation of xylanase by Penicillium janthinellum in a poly(ethylene glycol)/cashew-nut tree gum aqueous two-phase system; Oliveira LA et al.; Cultivation of the fungus Penicillium janthinellum for xylanase production was studied in a poly(ethylene glycol)/cashew-nut tree gum aqueous two-phase system, using a two-level fractional factorial design . The parameters studied were initial pH, cultivation time, type of agro-industrial residue (oat husk or corn cob), agitation, temperature, and phase-forming polymers . The xylanase produced during fermentation partitioned into the top phase . The agitation and temperature (negative), cultivation time and initial pH (positive) effects proved statistically significant for xylanase production . The highest percentage yield of the xylanase in the top and its production in the top phase, about 97% and 160.7 U/mL, were obtained in cultures of 120 h, 40 rpm, 25 degrees C, and pH 5.0. Biotechnol Prog, 2004 Nov-Dec, 20(6), 1641 - 50 Xylose metabolism in Debaryomyces hansenii UFV-170 . Effect of the specific oxygen uptake rate; Sampaio FC et al.; The new yeast Debaryomyces hansenii UFV-170 was tested in this work in batch experiments under variable oxygenation conditions . To get additional information on its fermentative metabolism, a stoichiometric network was proposed and checked through a bioenergetic study performed using the experimental data of product and substrate concentrations . The yeast metabolism resulted to be practically inactive under strict oxygen-limited conditions (qO2 = 12.0 mmol(O2) C-mol(DM)(-1) h(-1)), as expected by the impossibility of regenerating NADH2+ . Significant fractions of the carbon source were addressed to both respiration and biomass growth under excess oxygen levels (qO2 > or = 55.0 mmol(O2) C-mol(DM)(-1) h(-1)), thus affecting xylitol yield (Y(P/S) = 0.41-0.52 g g(-1)) . Semi-aerobic conditions (qO2 = 26.8 mmol(O2) C-mol(DM)(-1) h(-1)) were able to ensure the best xylitol production performance (Pmax = 76.6 g L(-1)), minimizing the fractions of the carbon source addressed either to respiration or biomass production and increasing Y(P/S) up to 0.73 g g(-1) . An average P/O ratio of about 1.0 mol(ATP) mol(O)(-1) allowed estimation of the main kinetic-bioenergetic parameters of the biosystem . The overall ATP requirements of biomass were found to be particularly high and dependent on the oxygen availability in the medium as well as on the physiological state of the culture . Under semi-aerobic and aerobic conditions, they varied in the ranges 13.5-15.4 and 9.74-10.2 mol(ATP) C-mol(DM)(-1), respectively, whereas during the best semi-aerobic bioconversion they progressively increased from 5.68 to 24.7 mol(ATP) C-mol(DM)(-1) . After a starting phase of adaptation to the medium, the cell achieved a phase of decelerated growth during which its excellent xylose-to-xylitol capacity kept almost constant after 112 h up to the end of the run. Appl Environ Microbiol, 2004 Dec, 70(12), 7520 - 9 Isolation and characterization of novel marine-derived actinomycete taxa rich in bioactive metabolites; Magarvey NA et al.; A unique selective enrichment procedure has resulted in the isolation and identification of two new genera of marine-derived actinobacteria . Approximately 90% of the microorganisms cultured by using the presented method were from the prospective new genera, a result indicative of its high selectivity . In this study, 102 actinomycetes were isolated from subtidal marine sediments collected from the Bismarck Sea and the Solomon Sea off the coast of Papua New Guinea . A combination of physiological parameters, chemotaxonomic characteristics, distinguishing 16S rRNA gene sequences, and phylogenetic analysis based on 16S rRNA genes provided strong evidence for the two new genera (represented by strains of the PNG1 clade and strain UMM518) within the family Micromonosporaceae . Biological activity testing of fermentation products from the new marine-derived actinomycetes revealed that several had activities against multidrug-resistant gram-positive pathogens, malignant cells, and vaccinia virus replication. Appl Environ Microbiol, 2004 Dec, 70(12), 7210 - 9 Allelic diversity and population structure in Oenococcus oeni as determined from sequence analysis of housekeeping genes; de Las Rivas B et al.; Oenococcus oeni is the organism of choice for promoting malolactic fermentation in wine . The population biology of O . oeni is poorly understood and remains unclear . For a better understanding of the mode of genetic variation within this species, we investigated by using multilocus sequence typing (MLST) with the gyrB, pgm, ddl, recP, and mleA genes the genetic diversity and genetic relationships among 18 O . oeni strains isolated in various years from wines of the United States, France, Germany, Spain, and Italy . These strains have also been characterized by ribotyping and restriction fragment length polymorphism (RFLP) analysis of the PCR-amplified 16S-23S rRNA gene intergenic spacer region (ISR) . Ribotyping grouped the strains into two groups; however, the RFLP analysis of the ISRs showed no differences in the strains analyzed . In contrast, MLST in oenococci had a good discriminatory ability, and we have found a higher genetic diversity than indicated by ribotyping analysis . All sequence types were represented by a single strain, and all the strains could be distinguished from each other because they had unique combinations of alleles . Strains assumed to be identical showed the same sequence type . Phylogenetic analyses indicated a panmictic population structure in O . oeni . Sequences were analyzed for evidence of recombination by split decomposition analysis and analysis of clustered polymorphisms . All results indicated that recombination plays a major role in creating the genetic heterogeneity of O . oeni . A low standardized index of association value indicated that the O . oeni genes analyzed are close to linkage equilibrium . This study constitutes the first step in the development of an MLST method for O . oeni and the first example of the application of MLST to a nonpathogenic food production bacteria. Front Biosci, 2005 Jan 1, 10, 99 - 106 Print 2005 Jan 1. Biosensor commercialization strategy - a theoretical approach; Lin CT et al.; Biosensors are analytical devices, which use biological interactions to provide either qualitative or quantitative results . They are extensively employed in many fields such as clinical diagnosis and biomedicine, military applications, anti-terrorism, farm, garden and veterinary analysis, process control, fermentation control and analysis, pharmaceutical and drug analysis, food and drink production and analysis, pollution control and monitoring, microbiology, bacterial and viral analysis, mining, and industrial and toxic gases . The biosensor market has significantly increased and will be mushrooming in the next decade . The total biosensor market is estimated to be $10.8 billion by 2007 . The emerging biosensor market presents both opportunities and obstacles to start-up biosensor entrepreneurs . The major challenge and threat for these entrepreneurs is how to predict the biosensor market and how to convert promising biosensor technology into commercialized biosensors . By adopting a simple commercialization strategy framework, we identify two key elements of biosensor commercialization strategy: excludability and complementary asset . We further divide biosensor commercialization environments into four distinct sub-environments: the Attacker's Advantage, Reputation-Based Idea Trading, Greenfield Competition and Ideas Factories . This paper explains how the interaction between these two key elements shapes biosensor commercialization strategy and biosensor industry dynamics . This paper also discusses alternative commercialization strategies for each specific commercialization environment and how to choose from these alternatives . The analysis of this study further provides a good reference for start-up biosensor entrepreneurs to formulate effective biosensor commercialization strategy. Front Biosci, 2005 Jan 1, 10, 94 - 8 Print 2005 Jan 1. Effect of enteric flora on inflammatory and angiogenic mechanisms in human intestinal microvascular endothelial cells; Ogawa H et al.; The intestine is a highly vascularized organ, and the splanchnic microcirculation is now appreciated to play a key role in immune and inflammatory responses in the gut . Emerging evidence demonstrates that the enteric flora not only exerts an important effect on innate immunity and the mucosal immune system, but will also affect inflammatory and angiogenic mechanisms involving the gut microcirculation . In response to bacterial lipopolysaccharide, the intestinal microcirculation and its endothelial lining will undergo activation, which will contribute to cell adhesion molecule expression and the recruitment of leukocytes into the gut wall, an early and rate limiting step in the inflammatory process . This is balanced by the fact that human intestinal microvascular endothelial cells possess specific mechanisms of endotoxin tolerance, which will diminish inflammatory activation in response to repeated bacterial activation . Likewise, the process of angiogenesis, or new blood vessel growth is influenced by the presence of bacteria, which stimulate the release of angiogenic factors from innate immune mechanisms . Conversely, bacterial derived products of dietary carbohydrate fermentation, the short chain fatty acids, will decrease angiogenic mechanisms in human intestinal microvascular endothelial cells . In this review we summarize our present state of knowledge regarding the interplay between enteric flora and inflammatory and angiogenic activation of the intestinal microcirculation its microvascular endothelium. Environ Sci Technol, 2004 Nov 15, 38(22), 5902 - 14 Stimulated microbial reductive dechlorination following surfactant treatment at the Bachman Road site; Ramsburg CA et al.; A pilot-scale demonstration of surfactant-enhanced aquifer remediation (SEAR) was conducted in July 2000 at the Bachman Road site located in Oscoda, MI . The Bachman aquifer is a shallow, relatively homogeneous, unconfined aquifer formation composed primarily of sandy glacial outwash with relatively low organic carbon content (0.02 wt %) . A 6 wt % aqueous solution of Tween 80 (a nonionic, food-grade surfactant) was flushed through a localized dense nonaqueous phase liquid (DNAPL) source zone to recover approximately 19 L of tetrachloroethene (PCE) . Post-treatment monitoring revealed PCE concentrations were reduced by up to 2 orders of magnitude within the source zone, and there was no evidence of concentration rebound after more than 450 d . Concentrations of PCE dechlorination products (trichloroethene, cis-1,2-dichloroethene) 450 d after SEAR operations ceased were more than 2 orders of magnitude greater than pretreatment values, suggesting stimulation of native dechlorination activity . Post-treatment monitoring detected increased concentrations of volatile fatty acids generated from the fermentation of residual-level Tween 80 surfactant . These field data suggest that Tween 80 not only induced and maintained anaerobiosis but also provided reducing equivalents to reductively dechlorinating populations present in the oligotrophic Bachman aquifer . Experience from this site supports application of staged treatment strategies that couple SEAR and microbial reductive dechlorination to enhance mass removal and reduce contaminant mass flux emanating from treated source zones. Indian J Exp Biol, 2004 Aug, 42(8), 837 - 43 Improvement of dry matter digestibility of water hyacinth by solid state fermentation using white rot fungi; Mukherjee R et al.; Feeding value of water hyacinth biomass colonized by three species of white rot fungi during solid-state fermentation was investigated . All three organisms proved to be efficient degraders and enhanced dry matter digestibility . Loss of organic matter was maximum (23.6+/-0.1% dry wt) after 48 days by P . ostreatus . C . indica showed maximum cellulose degradation (18.5+/-0.1% dry wt) than other two fungi after 48 days of incubation . In all cases, an extensive removal of hemicellulose at the initial growth period and a delayed degradation of lignin were observed . Hemicellulolysis was maximum (46.3+/-0.1% dry wt) by C . indica, but delignification (14.2+/-0.2% dry wt) by P . sajor-caju after 48 days . The amount of reducing sugar in the degraded biomass decreased at early stages, but increased as degradation progressed in all three cases (maximum 1.1+/-0.05% dry wt after 48 days by C . indica) . Soluble nitrogen content increased only during 16-32 days of incubation (highest 1.1+/-0.1% dry wt after 32 days by P . sajor-caju) . Crude protein of the bioconverted biomass increased gradually up to 32 days but decreased thereafter (maximum 10.3+/-0.1% dry wt after 32 days by P . sajor - caju) . Per cent change in in vitro dry matter digestibility of degraded substrates enhanced gradually after 8 days and reached maximum after 32 days but thereafter decreased (highest + 20.4+/-0.3% dry wt by P . sajor-caju) . The results demonstrated the efficient degrading capacity of the test fungi and their potential use in conversion of water hyacinth biomass into mycoprotein-rich ruminant feed, more so by P . sajor-caju. Arch Anim Nutr, 2004 Aug, 58(4), 325 - 42 Digesta characteristics of dorsal, middle and ventral rumen of cows fed with different hay qualities and concentrate levels; Tafaj M et al.; The influence of fibre content of hay (H) and concentrate level (C) on local differences in the composition of ruminal digesta (ratio of solid to fluid digesta, DM, NDF, ADF and ADL content), particle size (MPL), specific gravity (SG) and fermentation (pH and concentrations of SCFA and bicarbonate) have been tested on two ruminally cannulated Friesian cows (520 kg BW) which were fed restricted, using individual cows as experimental units . Digesta samples were collected via cannula from three rumen layers: 5 to 10 cm (top) and 25-35 cm beneath the top of the particle mat (middle) and 5-10 cm above the rumen floor (bottom) . For a main plot treatment (H x C), repeated samples were collected at four time intervals (1 h before and 2, 5 and 10 h after morning feeding) on each of two days . From top to bottom rumen the share of solid digesta mass (SM), DM and NDF contents of squeezed digesta fluid (SRF) and concentration of SCFA decreased (P < 0.05); pH and bicarbonate concentration increased (P < 0.05), while DM, NDF, ADF and ADL contents in SM, MPL and SG did not differ . Higher NDF content of hay (from 47-62%) increased SM, fibre fractions in SM, MPL, pH and concentration of bicarbonate in ruminal digesta, especially when 50% concentrate was given, while SG decreased . When the concentrate level was enhanced from 20 to 50%, digesta SM, MPL and the content of DM and NDF in SRF increased, while pH, concentrations of SCFA and acetate decreased when low-fibre hay was given . With longer time after feeding the digesta SM was reduced and fibre content in SM increased . The increase of the fibre content of hay reduced the possible negative effect of high concentrate level on the stratification of ruminal digesta . The decrease of the fibre content of hay promised better conditions for fibre digestion in the rumen when concentrate availability is limited. J Nutr, 2004 Dec, 134(12), 3233 - 8 An increase in reactive oxygen species by dietary fish oil coupled with the attenuation of antioxidant defenses by dietary pectin enhances rat colonocyte apoptosis; Sanders LM et al.; We showed previously that the dietary combination of fish oil, rich in (n-3) fatty acids, and the fermentable fiber pectin enhances colonocyte apoptosis in a rat model of experimentally induced colon cancer . In this study, we propose that the mechanism by which this dietary combination heightens apoptosis is via modulation of the colonocyte redox environment . Male Sprague-Dawley rats (n = 60) were fed 1 of 2 fats (corn oil or fish oil) and 1 of 2 fibers (cellulose or pectin) for 2 wk before determination of reactive oxygen species (ROS), oxidative DNA damage, antioxidant enzyme activity {superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx)} and apoptosis in isolated colonocytes . Fish oil enhanced ROS, whereas the combination of fish oil and pectin suppressed SOD and CAT and enhanced the SOD/CAT ratio compared with a corn oil and cellulose diet . Despite this modulation to a seemingly prooxidant environment, oxidative DNA damage was inversely related to ROS in the fish oil and pectin diet, and apoptosis was enhanced relative to other diets . Furthermore, apoptosis increased exponentially as ROS increased . These results suggest that the enhancement of apoptosis associated with fish oil and pectin feeding may be due to a modulation of the redox environment that promotes ROS-mediated apoptosis. Ultrasonics, 2005 Feb, 43(4), 247 - 52 Ultrasonic velocity in water-ethanol-sucrose mixtures during alcoholic fermentation; Resa P et al.; During alcoholic fermentation, sucrose and water are transformed into ethanol and carbon dioxide by the action of yeast enzymes . The measurement of the velocity of an ultrasonic pulse travelling through a fermentation tank can be used to characterize the state of the process . In this work, an experimental study of the density and ultrasonic velocity in the ternary mixture (water-ethanol-saccharose) is presented . Experimental results were compared to ideal density and to commonly used expressions of the sound velocity in liquid mixtures (Urick, Natta-Baccaredda and Nomoto) . A semiempirical approach was proposed to improve the efficiency of theoretical models when dealing with mixtures of associated liquids. Water Sci Technol, 2004, 50(8), 209 - 16 Effects of substrate components on hydrogen fermentation of multiple substrates; Bai MD et al.; As is well known, carbohydrate is the most appropriate organic material for hydrogen fermentation, and its hydrogen yield is significantly larger than that of protein . The fermentation of protein began with hydrogen production followed by hydrogen consumption, which helps overall hydrogen recovery . Both carbohydrate and protein are basic components of organic material, and yet carbohydrate is known to be a better substrate than protein in terms of hydrogen yield during hydrogen fermentation . This study used multiple substrates containing different ratios of glucose and peptone as multiple substrates to investigate the roles played by carbohydrate and protein in hydrogen fermentation . The experimental results demonstrated that suitable ratios of glucose and peptone improved the growth of hydrogen producing bacteria . Additionally, a maximum hydrogen yield of 6.4 mmole-H2/g-COD was obtained from the multiple substrate containing 40% peptone and 60% glucose . Most of the produced hydrogen came from fermentation of glucose, not peptone . During hydrogen fermentation, the pH dropped by 1.0 and 1.9 units in 80% and 20% of peptone content in the substrate . Ammonia produced due to peptone degradation neutralized the acids produced from hydrogen fermentation. Yeast, 2004 Dec, 21(16), 1375 - 89 A laboratory yeast strain suitable for spirit production; Schehl B et al.; Yeast strains of the species Saccharomyces cerevisiae currently in use for the production of consumable alcohols such as beer, wine and spirits are genetically largely undefined . This prevents the use of standard genetic manipulations, such as crossings and tetrad analysis, for strain improvement . Furthermore, it complicates the application of the majority of modern methods developed in yeast molecular biology . Here we used two haploid laboratory strains with suitable auxotrophic markers for the construction of a genetically well defined, prototrophic diploid production strain . This strain was tested for its fermentative and sensory performances in comparison to commercially available yeasts . Three different fruit mashes (cherries, plums and pears) were fermented in a 90 kg scale . These were then subjected to distillation and used for the production of spirits with a final ethanol content of 40% (v/v) . Fermentation parameters assayed included growth, sugar utilization, ethanol production and generation of volatile compounds, higher alcohols and glycerol . The spirits were also tested for their sensory performances and the data obtained statistically consolidated . Our results clearly demonstrate that this laboratory strain does not display any disadvantage compared with commercial yeasts in spirit production for any of the parameters tested, yet it offers the potential to apply both classical breeding and modern molecular genetic techniques for adjusting yeast physiology to special production schemes. Biosci Biotechnol Biochem, 2004 Nov, 68(11), 2401 - 4 Increased levels of policosanol and very long-chain fatty acids in potato pulp fermented with Rhizopus oryzae; Musa R et al.; Significant amounts of policosanol and very long-chain fatty acids (VLFAs) ranging in carbon length from 22 to 30 were found in the lipophilic fraction obtained from potato pulp fermented with Rhizopus oryzae . It is believed that these compounds would have originally been present as suberin-related compounds, but not as wax, in the periderm of potato tubers and concentrated into potato pulp during the process of starch production . Moreover, the policosanol and VLFAs extracted from potato pulp with organic solvents were found to have increased after fermentation. Biosci Biotechnol Biochem, 2004 Nov, 68(11), 2239 - 46 Caffeoylsophorose, a new natural alpha-glucosidase inhibitor, from red vinegar by fermented purple-fleshed sweet potato; Matsui T et al.; The suppressive effect on the postprandial blood glucose rise through alpha-glucosidase (AGH) inhibition was investigated in this study in order to clarify an antihyperglycemic function of 6-O-caffeoylsophorose (CS) from diacylated anthocyanin . The administration of CS (100 mg/kg) following maltose (2 g/kg) to Sprague-Dawley rats resulted in the maximal blood glucose level after 30 min being significantly decreased by 11.1% compared to the control . A reduction in the serum insulin secretion was also observed in parallel to the decrease in blood glucose level . No blood glucose change was apparent when sucrose or glucose was ingested, suggesting that the antihyperglycemic effect of CS was achieved by maltase inhibition, rather than by sucrase or glucose transport inhibition . An AGH inhibitory assay demonstrated that the non-competitive maltase inhibition of CS was partly due to acylation by phenolic acid with sugar, the presence of hydroxyl groups in the aromatic ring, and the presence of an unsaturated alkyl chain in the acylated moiety. J Agric Food Chem, 2004 Dec 1, 52(24), 7324 - 30 Vitamin content and amino acid composition of pickled garlic processed with and without fermentation; Montano A et al.; The effect of processing, with and without fermentation, upon the nutritional composition of pickled garlic was evaluated . On a dry basis, the fermented product had a higher content of riboflavin, alpha-tocopherol, and most individual amino acids but a lower thiamin level than the unfermented product . Ascorbic acid was totally lost during processing . The chemical scores for the unfermented and fermented product were 88 and 108%, respectively, with the limiting amino acid being leucine . Water blanching (90 degrees C for 4 min) affected only the ascorbic acid content, whereas fermentation significantly affected the contents of thiamin, ascorbic acid, and alpha-tocopherol, as well as glutamic acid and arginine . For each processing type, the effect of the preservation method and storage time on vitamins and amino acid composition was also analyzed . In the case of the fermented product, usage of the corresponding fermentation brine plus refrigerated storage was also assayed as the packing/preservation method and was found to give the best result from a nutritional standpoint. Biodegradation, 2004 Dec, 15(6), 395 - 403 Complete biological dehalogenation of chlorinated ethylenes in sulfate containing groundwater; Hoelen TP et al.; The ability of dehalogenating bacteria to compete with sulfate reducing bacteria for electron donor was studied in microcosms that simulated groundwater contaminated with both chlorinated ethylenes and fuel hydrocarbon compounds . Results demonstrate that reductive dehalogenation of perchloroethylene to ethylene can proceed in the presence of > 100 mg l(-1) sulfate . The hydrogen concentration, which was 2.5 nM in the presence of approximately 150 mg l(-1) sulfate and in the absence of chlorinated compounds, decreased to 0.7 nM during the dechlorination of trichloroethylene and increased to 1.6 nM during the dechlorination of cis-dichloroethylene and vinyl chloride . With only sediment associated donor ("historical" donor) present, dechlorination of trichloroethylene proceeded slowly to ethylene (on a time scale of several years) . Addition of toluene, a model hydrocarbon compound, stimulated dechlorination indirectly . Toluene degradation was rapid and linked to sulfate utilization, and presumably formed fermentable substrates that served as hydrogen donors . Dehalogenation was inhibited in soil free microcosms containing 5 mM sulfide, but inhibition was not observed when either aquifer sediment or 5 mM ferrous chloride was added. J Public Health Dent, 2004 Fall, 64(4), 244 - 8 Pediatricians' perception about the use of antibiotics and dental caries--a preliminary study; da Silva Pierro VS et al.; OBJECTIVES: This study aimed to evaluate pediatricians' perception about the use of antibiotics and dental caries . METHODS: One hundred pretested questionnaires containing open and closed questions were distributed to pediatricians from public hospitals in Rio de Janeiro, Brazil . RESULTS: The response rate was 60 percent . Considering the respondents, 73.3 percent frequently prescribed antibiotics, oral administration being the chosen mode (93.3%) . Antibiotics were pointed out as being very sweet substances by 53.3 percent of the pediatricians . However, the same percentage did not know what sweetener was responsible for the sweet flavor, and just 40.6 percent recommended oral hygiene after the medicine's intake . Among the pediatricians, 56.7 percent related the use of antibiotics to the occurrence of dental effects, and 40 percent thought that antibiotics could cause a defect of tooth structure . Only one pediatrician (3.2%) associated the presence of fermentable carbohydrates in the composition of these medicines with dental caries . CONCLUSIONS: Pediatricians in this study did not perceive the correct relationship between the presence of fermentable carbohydrates in antibiotics and dental caries . Many of them believed these medicines promoted a defect of tooth structure favoring the development of dental caries . Further studies with a larger sample are necessary. Antimicrob Agents Chemother, 2004 Dec, 48(12), 4892 - 4 Antimycoplasmal activity of hydroxytyrosol; Furneri PM et al.; The aim of this study was to investigate the in vitro antimycoplasmal activity of hydroxytyrosol . Twenty strains of Mycoplasma hominis, three strains of Mycoplasma fermentans, and one strain of Mycoplasma pneumoniae were used . For M . pneumoniae, M . hominis, and M . fermentans, the MICs were 0.5, 0.03 (for 90% of the strains tested), and 0.25 microg/ml, respectively. Nutrition, 2004 Nov-Dec, 20(11-12), 979 - 83 Bioavailability of cellobiose by tolerance test and breath hydrogen excretion in humans; Nakamura S et al.; OBJECTIVE: Prebiotic substances have the property of intestinal fermentation . Cellobiose has a beta-1,4 linkage, so it is resistant to hydrolysis by human small intestinal disaccharidase and, hence, reaches the colon undigested . Until this study, it was unclear whether cellobiose has fermentability or bioavailability . The objectives of this study were to clarify whether cellobiose is fermented in the large intestine and to estimate the available energy from cellobiose intake by using tolerance tests and breath hydrogen tests in healthy female subjects . METHODS: Ten healthy young women (20.5 +/- 2.1 y) who did not develop diarrhea after ingesting 30 g of cellobiose in a previous experiment were recruited . Tolerance tests and breath hydrogen tests for 25 g of cellobiose or glucose were carried out at least 2 wk apart . Blood samples were collected before and at 30-min intervals up to 3 h after ingestion . Breath gas samples were collected simultaneously before and at 30-min intervals up to 6 h after ingestion of cellobiose or glucose . Blood glucose and insulin levels and the concentration of breath hydrogen were analyzed . RESULTS: When 25 g of cellobiose was ingested, there was no increase in blood glucose or insulin secretion, but these markers increased remarkably with glucose ingestion . The excretion of breath hydrogen gas after cellobiose ingestion was significantly greater than that after glucose ingestion . CONCLUSIONS: Orally ingested cellobiose was well fermented in human large intestine, and its available energy was estimated to be about 2 kcal/g. J Environ Sci (China), 2004, 16(5), 816 - 20 Optimization of solid fermentation of cellulase from Trichoderma koningii; Li PJ et al.; To exploit peashrub resources in Ordos as fodders, it is very crucial to realize industrial production of cheap cellulase of high activity by optimizing culture technology, especially culture substrate . In this study, a new prescription experiment based on uniform design ideal was invented and successfully applied in the solid fermentation of Trichoderma koningii F244, which was performed with two different temperature degrees . The activities of FPA, cotton lyase, CMCase and beta-glucosidase were assayed and then mathematical models of enzymatic activities, which were figured out by Unconstraint Mathematical Programming, were developed by Multivariate Regression Program of SPSS10.0 . Enzymatic activities of optimized substrate prescriptions corresponding to mathematical models were forecasted to determine an ideal substrate prescription . It is revealed that in solid fermentation, Tween80 has negative effect on cellulase production . Furthermore, the ideal prescription for cellulase complex production by Trichoderma koningii F244 was straw powder 16.9%, wheat bran 26.5%, (NH4)2SO4 9.5% and water 47.1%, whose corresponding cellulase activity was expected to be at the same high level with that of Trichoderma reesei Q9414 on its own recommended substrate . Especially, goats mainly fed on peashrub tissues mixed with cellulase complex of this prescription and culture technology, got an incremental ratio of 0.3 kg/d, which brought a very promising feeding prospect for local peashrub resource . By populization of this cellulase complex, it can integrate living standard, economic construction of local residents into vegetational restoration tightly and thus this paper will be very meaningful to be use for reference for western China like Ordos to realize its sustainable development of economy, society and environment. J Basic Microbiol, 2004, 44(6), 471 - 9 Species identification and comparative molecular and physiological analysis of Candida zemplinina and Candida stellata; Sipiczki M; Strains of the recently described yeast species Candida zemplinina have been identified in botrytized grapes and musts in geographically distant areas . C . zemplinina is almost indistinguishable from C . stellata, another yeast common in fermenting botrytized wines . In this work a specific PCR-RFLP method is described for the differentiation of the two species . It is also shown that both species have three chromosomes, display chromosomal length polymorphism, and C . zemplinina has a smaller genome . C . zemplinina is acidogenic, highly osmotolerant, grows much better than C . stellata in the presence of ethanol and at low temperatures . ((c) 2004 WILEY-VCH Verlag GmbH & Co . KGaA, Weinheim). Comp Biochem Physiol A Mol Integr Physiol, 2004 Nov, 139(3), 293 - 300 Nutritional and sanitary statuses alter postweaning development of caecal microbial activity in the rabbit; Bennegadi-Laurent N et al.; The postweaning development of caecal microbial activity was studied in the rabbit according to the sanitary status (conventional "C" vs . specified pathogen-free "SPF") and the nutritional status (standard-fibre "SF" vs . deficient-fibre "DF" diet) . The two diets were distributed ad libitum from weaning (28 days) to 70 days of age, respectively, to 80 C and 72 SPF rabbits . From 28 to 42 days, the volatile fatty acids concentration in the caecum (tVFA) of C rabbits was 50 mM/L and increased by 46% between 42 and 56 days, without interactions with the diet effect . In parallel, the bacterial fibrolytic activity decreased for xylanase and CMCase (-32% and -60%, respectively, P<0.05), while pectinase activity decreased more regularly from 28 to 70 days (-28%, P<0.05) . At weaning, tVFA was similar among C or SPF rabbits, while at 70 days, it decreased by 23% for SPF and increased in C group (+31%) . Cellulasic and hemicellulasic activity of bacteria were two to three times lower, respectively, in SPF rabbits compared to conventional ones . No interaction was detected between sanitary and nutritional status at 70 days of age for the caecal fermentative activity . With the FD diet, tVFA decreased by 10%, while butyrate proportion increased by 37% (at 70 days), whatever the sanitary status . In 70-day-old rabbits (C or SPF group), pectinasic activity was reduced by 30% when rabbits were fed the FD compared to the SF one. FEMS Yeast Res, 2004 Dec, 5(3), 237 - 45 Enological and genetic traits of Saccharomyces cerevisiae isolated from former and modern wineries; Cocolin L et al.; Saccharomyces cerevisiae strains isolated from two different wineries in central Italy were subjected to enological and molecular characterization to investigate the influence of the winery environment . One of the selected wineries is a modern, working winery, whereas the second one was abandoned since 1914 and was located in an artificial cavern . The results obtained by our analysis of the fermentation traits underline the selectivity of the winery environment (winery effect), since strains isolated from the industrial winery showed higher values for characters typically subjected to selective pressure, such as maximum capability to produce ethanol, fermentation rate and SO(2) resistance . Pulsed-field gel electrophoresis (PFGE), random amplification of polymorphic DNA (RAPD)-PCR and SAU-PCR were carried out to assesss genetic differences between the two populations studied . Only RAPD-PCR could distinguish between the two populations based on their provenience, whereas PFGE and SAU-PCR gave profiles shared between strains isolated from the industrial and former winery . Moreover, analysis of the karyotypes suggested the presence of chromosomal-length polymorphism; differences in the size and number of chromosomes between the two groups of isolates, as well as within each group, were observed. FEMS Yeast Res, 2004 Dec, 5(3), 213 - 30 Two-dimensional protein map of an "ale"-brewing yeast strain: proteome dynamics during fermentation; Kobi D et al.; The first protein map of an ale-fermenting yeast is presented in this paper: 205 spots corresponding to 133 different proteins were identified . Comparison of the proteome of this ale strain with a lager brewing yeast and the Saccharomyces cerevisiae strain S288c confirmed that this ale strain is much closer to S288c than the lager strain at the proteome level . The dynamics of the ale-brewing yeast proteome during production-scale fermentation was analysed at the beginning and end of the first and the third usage of the yeast (called generation in the brewing industry) . During the first generation, most changes were related to the switch from aerobic propagation to anaerobic fermentation . Fewer changes were observed during the third generation but certain stress-response proteins such as Hsp26p, Ssa4p and Pnc1p exhibited constitutive expression in subsequent generations . The ale brewing yeast strain appears to be quite well adapted to fermentation conditions and stresses. Protein Expr Purif, 2004 Dec, 38(2), 264 - 71 High level recombinant protein expression in Ralstonia eutropha using T7 RNA polymerase based amplification; Barnard GC et al.; We report further development of a novel recombinant protein expression system based on the Gram-negative bacterium, Ralstonia eutropha . In this study, we were able to express soluble, active, organophosphohydrolase (OPH), a protein that is prone to inclusion body formation in Escherichia coli, at titers greater than 10 g/L in high cell density fermentation . This represents a titer that is approximately 100-fold greater than titers previously reported in E . coli for this enzyme . R . eutropha strains expressing OPH were generated in two cloning steps . First, the T7 RNA polymerase gene was placed under the control of the strong, inducible phaP promoter and integrated into the phaP locus of R . eutropha NCIMB 40124 . Second, a single copy of the oph gene under control of the T7 promoter was randomly integrated into the chromosome using a transposon cloning vector. Mol Microbiol, 2004 Dec, 54(5), 1326 - 34 Positive control of tylosin biosynthesis: pivotal role of TylR; Stratigopoulos G et al.; Control of tylosin production in Streptomyces fradiae features interplay between a repressor, TylQ, and an activator, TylS, during regulation of tylR . The latter encodes a pathway-specific activator that controls most of the tylosin-biosynthetic (tyl) genes that are subject to regulation . This was established by targeted gene disruption applied separately to tylR and tylS together with transcript analysis involving reverse transcription polymerase chain reaction (RT-PCR) . TylR controls multiple genes that encode the synthesis or addition of all three tylosin sugars, plus polyketide ring oxidation, and at least one of the polyketide synthase (PKS) megagenes, tylGI . (Expression of a few tyl genes, plus the resistance determinants tlrB and tlrD, together with some ancillary or unassigned genes, is not apparently regulated during fermentation, consistent with constitutive expression.) In contrast, the only gene known for sure to be directly controlled by TylS is tylR, and there are very few additional candidates . These include the mycinose-biosynthetic gene, tylJ, and two previously unassigned genes, ORF12* (tylU) plus ORF11* (tylV) . TylS also controls the PKS genes {tylGIII-tylGIV-tylGV} although not in obligatory fashion . These genes can be transcribed (i.e . tylosin can be produced) in a tylS-KO strain by forcing overexpression of tylR using a foreign promoter . We therefore suspect that TylS might control the PKS genes indirectly, although this remains to be established unequivocally . Conceivably, the direct effects of TylS are exerted exclusively on other regulators . Tylosin production levels were elevated when tylS or (especially) tylR was overexpressed in S . fradiae wild-type and yield increments of industrial significance were generated by similar manipulation of an enhanced production strain. Eur J Cancer Prev, 2004 Aug, 13(4), 345 - 8 Breast cancer chemopreventive properties of pomegranate (Punica granatum) fruit extracts in a mouse mammary organ culture; Mehta R et al.; We previously reported anticancer effects of pomegranate extracts in human breast cancer cells in vitro and also chemopreventive activity of pomegranate fermented juice polyphenols (W) in a mouse mammary organ culture (MMOC) . In the present study we decided to expand the MMOC investigations to also include an evaluation of the potential chemopreventive efficacy of a purified chromatographic peak of W (Peak B), and also of whole pomegranate seed oil . In brief, an MMOC was established according to a known method . For the first 10 days of culture, the glands were treated with pomegranate fermented juice polyphenols (W), a high-performance liquid chromatographic (HPLC) peak separated from W (peak B), or pomegranate seed oil (Oil, and on day 3, exposed to the carcinogen 7,12-dimethylbenz{a}anthracene (DMBA), and for 10 days treated with the putative pomegranate chemopreventive . The glands were subsequently harvested and tumours counted by visual inspection . While W effected a 42% reduction in the number of lesions compared with control, peak B and pomegranate seed oil each effected an 87% reduction . The results highlight enhanced breast cancer preventive potential both for the purified compound peak B and for pomegranate seed oil, both greater than that previously reported for pomegranate fermented juice polyphenols. Mikrobiol Z, 2004 Sep-Oct, 66(5), 76 - 83 {Extraction of a broad range of metals from sewage in the city of Kyiv by mixed microbe communities}; Protein engineering and applications of Candida rugosa lipase isoforms; Department of Food Science and Technology, University of Georgia, Athens, Georgia 30602-7610, USACommercial preparations of Candida rugosa lipase (CRL) are mixtures of lipase isoforms used for the hydrolysis and synthesis of various esters . The presence of variable isoforms and the amount of lipolytic protein in the crude lipase preparations lead to a lack of reproducibility of biocatalytic reactions . Purification of crude CRL improve their substrate specificity, enantioselectivity, stability, and specific activities . The expression of the isoforms is governed by culture or fermentation conditions . Unfortunately, the nonsporogenic yeast C . rugosa does not utilize the universal codon CTG for leucine; therefore, most of the CTG codons were converted to universal serine triplets by site-directed mutagenesis to gain expression of functional lipase in heterologous hosts . Recombinant expressions by multiple-site mutagenesis or complete synthesis of the lipase gene are other possible ways of obtaining pure and different CRL isoforms, in addition to culture engineering . Protein engineering of purified CRL isoforms allows the tailoring of enzyme function . This involves computer modeling based on available 3-D structures of lipase isoforms . Lid swapping and DNA shuffling techniques can be used to improve the enantioselectivity, thermostability, and substrate specificity of CRL isoforms and increase their biotechnological applications . Lid swapping can result in chimera proteins with new functions . The sequence of the lid can affect the activity and specificity of recombinant CRL isoforms . Candida rugosa lipase is toxicologically safe for food applications . Protein engineering through lid swapping and rationally designed site-directed mutagenesis will continue to lead to the production of CRL isoforms with improved catalytic power, thermostability, enantioselectivity, and substrate specificity, while providing evidence for the mechanisms of actions of the various isoforms. Poult Sci, 2004 Nov, 83(11), 1844 - 8 Testing the efficacy of fermented wheat germ extract against Mycoplasma gallisepticum infection of chickens; Stipkovits L et al.; The effect of fermented wheat germ extract (FWGE, Immunovet-HBM) was studied in chickens challenged with Mycoplasma gallisepticum . Ninety M . gallisepticum- and M . synoviae-free 3-wk-old chickens were exposed to aerosol infection of M . gallisepticum . One group (30 birds) was treated with FWGE, a second group with tiamulin, and a third group was untreated . The fourth group was exposed to PBS aerosol as a negative control . On d 9, all chickens were slaughtered and examined for the presence of gross and histological lesions, the presence of the challenge strain in the organs and specific antibodies in the serum . Body weight gains and feed conversion rates were recorded . In the groups treated with FWGE and with tiamulin, the chickens remained clinically healthy: their BW gains were 441.7 g and 446.8 g, respectively . Feed conversion ratios were 1.72 and 1.71 for FWGE- and tiamulin-treated birds, respectively . Control birds had BW gain of 480.8 g, and feed conversion ratio of 1.78 . The numbers of birds with gross lesions (15 and 11, respectively) and lesion scores (25 and 25, respectively) of the FWGE- and tiamulin-treated groups were significantly lower than in the infected untreated group (25 birds, lesion score of 190) . No mycoplasma was reisolated from brain, liver, spleen, heart, or kidneys of the FWGE-treated birds, and the number of mycoplasma isolations from the respiratory tract samples was less frequent (10) than from the infected untreated group (64) . In addition, 35 samples from other internal organs were also positive . Twenty percent of the birds treated with FWGE showed serological response with a 5.0% reaction score, whereas in the infected untreated group, 83.3% of birds were reactors, with a 62.5% reaction score. J Food Prot, 2004 Nov, 67(11), 2622 - 6 Detection of aflatoxin-producing molds in Korean fermented foods and grains by multiplex PCR; Yang ZY et al.; An assay based on multiplex PCR was applied for the detection of potential aflatoxin-producing molds in Korean fermented foods and grains . Three genes, avfA, omtA, and ver-1, coding for key enzymes in aflatoxin biosynthesis, were used as aflatoxin-detecting target genes in multiplex PCR . DNA extracted from Aspergillus flavus, Aspergillus parasiticus, Aspergillus oryzae, Aspergillus niger, Aspergillus terreus, Penicillium expansum, and Fusarium verticillioides was used as PCR template to test specificity of the multiplex PCR assay . Positive results were achieved only with DNA that was extracted from the aflatoxigenic molds A . flavus and A . parasiticus in all three primer pairs . This result was supported by aflatoxin detection with direct competitive enzyme-linked immunosorbent assay (DC-ELISA) . The PCR assay required just a few hours, enabling rapid and simultaneous detection of many samples at a low cost . A total of 22 Meju samples, 24 Doenjang samples, and 10 barley samples commercially obtained in Korea were analyzed . The DC-ELISA assay for aflatoxin detection gave negative results for all samples, whereas the PCR-based method gave positive results for 1 of 22 Meju samples and 2 of 10 barley samples . After incubation of the positive samples with malt extract agar, DC-ELISA also gave positive results for aflatoxin detection . All Doenjang samples were negative by multiplex PCR and DC-ELISA assay, suggesting that aflatoxin contamination and the presence of aflatoxin-producing molds in Doenjang are probably low. J Chromatogr A, 2004 Oct 29, 1054(1-2), 403 - 7 Determination of volatile N-nitrosamines in irradiated fermented sausage by gas chromatography coupled to a thermal energy analyzer; Byun MW et al.; Volatile N-nitrosodimethylamine (NDMA) and N-nitrosopyrrolidine (NPYR) in irradiated pepperoni and salami sausages were determined using a gas chromatography coupled to a thermal energy analyzer (GC-TEA) . These fermented sausages with aerobic or vacuum packaging were irradiated at 0, 5, 10, and 20 kGy, and then stored for 4 weeks at 4 degrees C . Both NDMA and NPYR in the fermented sausage were significantly reduced by irradiation . The vacuum packaging showed significantly lower (P < 0.05) N-nitrosamine levels than that of the aerobic ones . After storage, the contents of NDMA and NPYR in the irradiated sausage were lower than those of the non-irradiated control . Results indicated that a high dose of irradiation (>10 kGy) was needed to reduce the carcinogenic N-nitrosamines in the fermented sausage during storage and the GC-TEA analysis was effective in determining the N-nitrosamines in irradiated meats even at low trace levels. J Chromatogr A, 2004 Oct 29, 1054(1-2), 293 - 301 Characterization of winemaking yeast by cell number-size distribution analysis through flow field-flow fractionation with multi-wavelength turbidimetric detection; Zattoni A et al.; Yeasts are widely used in several areas of food industry, e.g . baking, beer brewing, and wine production . Interest in new analytical methods for quality control and characterization of yeast cells is thus increasing . The biophysical properties of yeast cells, among which cell size, are related to yeast cell capabilities to produce primary and secondary metabolites during the fermentation process . Biophysical properties of winemaking yeast strains can be screened by field-flow fractionation (FFF) . In this work we present the use of flow FFF (FlFFF) with turbidimetric multi-wavelength detection for the number-size distribution analysis of different commercial winemaking yeast varieties . The use of a diode-array detector allows to apply to dispersed samples like yeast cells the recently developed method for number-size (or mass-size) analysis in flow-assisted separation techniques . Results for six commercial winemaking yeast strains are compared with data obtained by a standard method for cell sizing (Coulter counter) . The method here proposed gives, at short analysis time, accurate information on the number of cells of a given size, and information on the total number of cells. Lett Appl Microbiol, 2004, 39(6), 504 - 8 Limitations of membrane cultures as a model solid-state fermentation system; Rahardjo YS et al.; AIMS: To examine the reliability of membrane cultures as a model solid-state fermentation (SSF) system . METHODS AND RESULTS: In overcultures of Aspergillus oryzae on sterilized wheat flour discs overlaid with a polycarbonate membrane, we demonstrated that the presence of membrane filters reduced the maximum respiration rate (up to 50%), and biomass and alpha-amylase production . We also show that the advantage of membrane cultures, i.e . total recovery of biomass, is not very evident for the system used, while the changes in metabolism and kinetics are serious drawbacks . CONCLUSIONS: The use of membrane cultures is artificial and without substantial benefits and therefore has to be carefully considered . SIGNIFICANCE AND IMPACT OF THE STUDY: In future studies on kinetics and stoichiometry of SSF, one should not completely rely on experiments using membrane cultures as a model SSF system. J Zhejiang Univ Sci, 2004 Dec, 5(12), 1597 - 603 Ant colony system algorithm for the optimization of beer fermentation control; Xiao J et al.; Beer fermentation is a dynamic process that must be guided along a temperature profile to obtain the desired results . Ant colony system algorithm was applied to optimize the kinetic model of this process . During a fixed period of fermentation time, a series of different temperature profiles of the mixture were constructed . An optimal one was chosen at last . Optimal temperature profile maximized the final ethanol production and minimized the byproducts concentration and spoilage risk . The satisfactory results obtained did not require much computation effort. J Zhejiang Univ Sci, 2004 Dec, 5(12), 1590 - 6 Scale-up of rifamycin B fermentation with Amycolatoposis mediterranei; Jin ZH et al.; Study of the effect of dissolved oxygen and shear stress on rifamycin B fermentation with A . mediterranei XC 9-25 showed that rifamycin B fermentation with Amycolatoposis mediterranei XC 9-25 needs high dissolved oxygen and is not very sensitive to shearing stress . The scale-up of rifamycin B fermentation with A . mediterranei XC 9-25 from a shaking flask to a 15 L fermentor was realized by controlling the dissolved oxygen to above 25% of saturation in the fermentation process, and the potency of rifamycin B fermentation in the 15 L fermentor reached 10 g/L after 6-day batch fermentation . By continuously feeding glucose and ammonia in the fermentation process, the potency of rifamycin B fermentaion in the 15 L fermentor reached 18.67 g/L, which was 86.65% higher than that of batch fermentation . Based on the scale-up principle of constantly aerated agitation power per unit volume, the scale-up of rifamycin B fed-batch fermentation with continuous feed from a 15 L fermentor to a 7 m(3) fermentor and further to a 60 m(3) fermentor was realized successfully . The potency of rifamycin B fermentation in the 7 m(3) fermentor and in the 60 m(3) fermentor reached 17.25 g/L and 19.11 g/L, respectively. Int J Oncol, 2004 Dec, 25(6), 1809 - 15 Preventive effect of fermented brown rice and rice bran on N-nitrosomethylbenzylamine-induced esophageal tumorigenesis in rats; Kuno T et al.; Modifying effect of fermented brown rice by Aspergillus Oryzae (FBRA) during the initiation or post-initiation phase of N-nitrosomethylbenzylamine (NMBA)-induced esophageal tumorigenesis was investigated in rats . Five-week-old male F344 rats were divided into 7 groups, and groups 1-5 were given subcutaneous injections of NMBA (0.5 mg/kg body weight/injection 15 times) for 5 weeks starting at 7 weeks of age . Groups 2 and 4 were fed the diet containing 5 and 10% FBRA during the initiation phase, respectively, whereas groups 3 and 5 were fed these diets during the post-initiation phase . Group 6 was given the diet containing 10% FBRA throughout the experim