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PubMed2005

Broomcorn millet grain cultures of the entomophthoralean fungus Zoophthora radicans: sporulation capacity and infectivity to Plutella xylostella.

The shelled grains of glutinous broomcorn millet, Panicum miliaceum, were used as solid substrate to prepare granular cultures of Zoophthora radicans, an entomophthoralean biocontrol agent against numerous insect pests. Steamed millet grains were inoculated by mixing 15 g millet grains (D.W.) with mashed pieces of half a 60-mm-dish colony in 3 ml modified Sabouraud dextrose broth and incubated at 15 degrees C and L:D 12:12 for up to 24 d. 20 grains were sampled at 3 d intervals from day six onwards and individually assessed for their sporulation capacity using a self-designed device for spore collection. The millet cultures after > or = 12 d incubation produced 12.0 - 14.9 x 10(4) spores grain(-1) during a 7 d period. The maximal sporulation capacity associated with the 21 d-old culture was about half of that of Z. radicans-killed Plutella xylostella larvae (28.7 x 10(4) spores cadaver(-1)), which individually were at least three times larger than the millet grains. Based on the time-concentration-mortality responses of second-instar P. xylostella larvae to Z. radicans in three independent bioassays, the spores ejected from the cultured millet grains, from the mycelial mats from liquid culture, and from larval cadavers displayed insignificant variations in infectivity to the host species, and yielded similar LC50 and LT50 estimates. Conclusively, the millet-based technology for production of granular cultures of Z. radicans was easy, inexpensive and highly efficient, and it could be superior to previous methods used in mass production of mycelium-based preparations of Entomophthorales since this new approach requires no special additives, drying, freezing and milling. This technology may suit to mass production of culturable but nutritionally fastidious entomopathogens from the Entomophthorales.

Animals↗
PubMed2005

Developmental competence and gene expression in preimplantation bovine embryos derived from somatic cell nuclear transfer using different donor cells.

This study compared the developmental competence of somatic cell nuclear transfer (SCNT) embryos reconstructed with different donor cells and analysed gene expression in the resulting embryos. Bovine fetal/adult ear fibroblasts and cumulus cells were used as donor cells and the developmental competence of the reconstructed embryos was monitored. The cell number and allocation in blastocysts were determined by differential staining. The Bax, E-cad, IF-tau, Hsp (heat shock protein) 70, Igf2r (insulin-like growth factor 2 receptor), DNMT (DNA methyltransferase) 1 and Mash (mammalian achaete-scute homologue) 2 genes were selected for gene expression analysis. The relative abundance (ratio to GAPDH mRNA) of gene transcripts in blastocysts was measured by semiquantitative reverse transcription-polymerase chain reaction. In experiment 1, development of SCNT preimplantation embryos and the cell numbers of inner cell masses and trophoblasts were not different among SCNT embryos derived from different cell types. In experiment 2, the relative expression of GAPDH and Hsp 70 transcripts was similar in all embryos. The expression of Bax, Igf2r and Mash2 transcripts was significantly increased in SCNT embryos reconstructed with adult fibroblasts. The E-cad transcript levels were reduced in SCNT embryos reconstructed with fetal fibroblasts. Relative abundance of DNMT1 in SCNT embryos derived from fetal fibroblasts was increased, and IF-tau expression in SCNT embryos derived from cumulus cells was increased. In conclusion, depending on the type of donor cells, preimplantation SCNT embryos displayed marked differences in gene expression. This may affect the developmental competence of SCNT embryos reconstructed with different cell types after implantation or during fetal growth in vivo.

Animals↗
PubMed2001

Evidence of the glycation and denaturation of LTP1 during the malting and brewing process.

The influence of malting and brewing processes on the chemical and structural modifications occurring on LTP1 was investigated by mass spectrometry and circular dichroism. Proteins were first purified from malt, and samples were collected at various steps of beer processing performed on two barley cultivars. The levels of LTP1 found in malt were not significantly different from the amounts in barley seed. However, in malt, both LTP1b, a post-translational form of LTP1, and a third isoform named LTP1c were isolated. Moreover, both of these proteins were found to be heterogeneously glycated but still exhibited an alpha-helix structure. Both glycated LTP1 and LTP1b were recovered during mashing. It was also shown that glycated LTP1 was unfolded during heat treatment of wort boiling, which is in agreement with the denatured form previously isolated from beer.

Beer↗
PubMed2001

Interactions of malt and barley (Hordeum vulgare L.) endoproteinases with their endogenous inhibitors.

For producing worts that are optimal for beer production, some, but not all, of the barley proteins must be degraded during malting and mashing. This protein hydrolysis is controlled by endoproteinases, and, in turn, is partially regulated by the presence of low-molecular-weight (LMW) proteinaceous inhibitors. This paper reports studies of the interactions between the proteinases and inhibitors and an "affinity" method for concentrating the inhibitors. The malt inhibitors (I) and proteinases (E) quickly formed strong (E-I) complexes when dissolved together, and all of the I was complexed. Heating at 100 degrees C, but not 70 degrees C, dissociated the complex, even though the enzyme activities were destroyed at 70 degrees C. The released I readily recomplexed with fresh E. Barley, however, contained insufficient E to complex all of its I complement. The E-I complex was treated with salts, detergents, and reducing agents to release active E molecules, but none disrupted the complex. By removing the LMW proteins from a malt E-I extract and dissociating the complex by heating, the concentration of I molecules was greatly increased. This "affinity" method can thus be used to concentrate the I molecules for further purification.

Beer↗
PubMed2003

Sulfite analysis of fruits and vegetables by high-performance liquid chromatography (HPLC) with ultraviolet spectrophotometric detection.

Free and total sulfite were analyzed in acidified vegetable products, instant mashed potatoes, and dried apples. Sulfite was separated by HPLC and quantified with a UV-vis detector. Resolution from components of food samples was achieved by varying the acid concentration of the eluant solution and by appropriate choice of the analytical wavelength. The minimum detectable levels for sulfite were 0.5 mg/L for a 10-cm analytical column and 1.5 mg/L for a 30-cm column. For analyses done with a 30-cm column, the coefficient of variation was <2% for analysis of free sulfite and total sulfite in acidified vegetables. For dried apples and instant potatoes, it ranged from 1 to 6.5%. The corresponding analytical errors were <4% and 1.2-5.6%, respectively, for the 10-cm column.

Chromatography, High Pressure Liquid↗
PubMed2004

Effect of technological processing on the allergenicity of mangoes (Mangifera indica L.).

In parallel with the rising popularity of exotic fruits in Europe, allergy against mango is of increasing importance. Because mangoes are also consumed as processed products such as chutneys or beverages, the influences of different process conditions on their allergenicity were investigated. Mango purees and nectars were manufactured at small pilot-plant scale, and the allergenic potencies of the resulting intermediate and final products were determined by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting and inhibitive enzyme allergosorbent tests (EAST-inhibition), using a pool serum of 9 individuals with manifest mango allergy. The mango allergens were shown to be very stable during technological processing. Irrespective of enzymatic matrix decomposition, mechanical tissue disintegration and heating during peeling, mash treatment, and pasteurization, significant loss of allergenicity could not be observed in the extracts of mango purees and nectars derived thereof. These results were confirmed by analogous investigation of commercial mango drinks and nectars. Hence, conventional mango processing into pulp-containing products typical for this species obviously does not allow complete elimination of the allergenic potency.

Allergens↗
PubMed2004

Ferulic acid release and 4-vinylguaiacol formation during brewing and fermentation: indications for feruloyl esterase activity in Saccharomyces cerevisiae.

The release of ferulic acid and the subsequent thermal or enzymatic decarboxylation to 4-vinylguaiacol are inherent to the beer production process. Phenolic, medicinal, or clove-like flavors originating from 4-vinylguaiacol frequently occur in beer made with wheat or wheat malt. To evaluate the release of ferulic acid and the transformation to 4-vinylguaiacol, beer was brewed with different proportions of barley malt, wheat, and wheat malt. Ferulic acid as well as 4-vinylguaiacol levels were determined by HPLC at several stages of the beer production process. During brewing, ferulic acid was released at the initial mashing phase, whereas moderate levels of 4-vinylguaiacol were formed by wort boiling. Higher levels of the phenolic flavor compound were produced during fermentations with brewery yeast strains of the Pof(+) phenotype. In beer made with barley malt, ferulic acid was mainly released during the brewing process. Conversely, 60-90% of ferulic acid in wheat or wheat malt beer was hydrolyzed during fermentation, causing higher 4-vinylguaiacol levels in these beers. As cereal enzymes are most likely inactivated during wort boiling, the additional release of ferulic acid during fermentation suggests the activity of feruloyl esterases produced by brewer's yeast.

Beer↗
PubMed2004

Distribution of ascorbic acid in potato tubers and in home-processed and commercial potato foods.

HPLC was used to analyze the content of ascorbic acid (AA) in tubers of four Korean potato cultivars (Chaju, Sumi, Deso, and Dejima), in a series of baked, boiled, braised, fried, microwaved, pressure-cooked, and sauteed potato slices from the Dejima cultivar and in 14 commercial Korean and 14 processed potato foods sold in the United States (chips, snacks, mashed potatoes, fries). The AA content for the four cultivars ranged from 16 to 46 mg/100 g of fresh weight. The distribution of AA in each of the eight potato slices (sticks, plugs) cut horizontally from the stem end of the Dejima potato ranged from 6.8 to 19.3% of the total. The corresponding distribution in seven sticks cut vertically was much narrower, ranging from 11.7 to 17.5% of the total. Losses of AA in water (pH 5.2) were significantly greater than in 5% metaphosphoric acid (pH 1.0). Less degradation occurred in water solutions of the vitamin stored at 1 degree C than at 25 degrees C. Losses of AA observed during home-processing of three varieties with low (Dejima, 16 mg/100 g), intermediate (Sumi, 32 mg/100 g), and high (Chaju, 42 mg/100 g) AA contents were as follows: boiling in water, 77-88%; boiling in water containing 1-3% NaCl, 61-79%; frying in oil, 55-79%; sauteing, 61-67%; pressure-cooking in water, 56-60%; braising, 50-63%; baking, 33-51%; and microwaving, 21-33%. The content of the Korean foods ranged from trace amounts to 25 mg/100 g and that of the U.S. foods from 0.4 to 46 mg/100 g. These results permit optimization of the vitamin C content of the diet by (a) using high-vitamin C potato varieties such as Chaju, (b) selecting sticks cut horizontally for frying, (c) baking or microwaving rather than boiling or frying, and (d) selecting commercial potato foods with a high vitamin C content.

Ascorbic Acid↗
PubMed2004

Improved sample preparation to determine acrylamide in difficult matrixes such as chocolate powder, cocoa, and coffee by liquid chromatography tandem mass spectroscopy.

An improved sample preparation (extraction and cleanup) is presented that enables the quantification of low levels of acrylamide in difficult matrixes, including soluble chocolate powder, cocoa, coffee, and coffee surrogate. Final analysis is done by isotope-dilution liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) using d3-acrylamide as internal standard. Sample pretreatment essentially encompasses (a) protein precipitation with Carrez I and II solutions, (b) extraction of the analyte into ethyl acetate, and (c) solid-phase extraction on a Multimode cartridge. The stability of acrylamide in final extracts and in certain commercial foods and beverages is also reported. This approach provided good performance in terms of linearity, accuracy and precision. Full validation was conducted in soluble chocolate powder, achieving a decision limit (CCalpha) and detection capability (CCbeta) of 9.2 and 12.5 microg/kg, respectively. The method was extended to the analysis of acrylamide in various foodstuffs such as mashed potatoes, crisp bread, and butter biscuit and cookies. Furthermore, the accuracy of the method is demonstrated by the results obtained in three inter-laboratory proficiency tests.

Acrylamide↗
PubMed2005

Behavior of myclobutanil, propiconazole, and nuarimol residues during lager beer brewing.

Over a 4 month brewing process, the fate of three fungicides, myclobutanil, propiconazole, and nuarimol, was studied in the spent grain, brewer wort, and final beer product. Only the residual level of myclobutanil after the mashing step was higher than its maximum residue limit (MRL) on barley. A substantial fraction was removed with the spent grain in all cases (26-42%). The half-life times obtained for the fungicides during storage of the spent grains ranged from 82 to 187 days. No significant influence of the boiling stage on the decrease of the fungicide residues was demonstrated. During fermentation, the content reduction varied from 20 to 47%. After the lagering and filtration steps, no significant decrease (<10%) was observed in any of the residues. Finally, during storage of the beer (3 months), the amounts of fungicides fell by 25-50% of their respective concentrations in the finished beer.

Beer↗
PubMed2006

Influence of the distillation step on the ratios of stable isotopes of ethanol in cherry brandies.

Isotope ratio mass spectrometry and site-specific natural isotope fractionation-nuclear magnetic resonance were applied to determine the overall carbon isotope ratio (delta13C) and the hydrogen isotope ratios [(D/H)I and (D/H)II] of ethanol, respectively. Ethanol was obtained by distillation of fermented cherry mash from a pot still commonly used in fruit brandy production. Analyses of distillate fractions revealed that the distillation proceeds with a fractionation of ethanol isotopologues. The inverse vapor pressure isotope effect (VPIE) observed for the carbon isotopologues is in accordance with the data reported for distillation of ethanol in spinning band columns. In contrast, the inverse VPIE for hydrogen isotopologues of ethanol observed in spinning band columns could not be confirmed. To investigate whether the observed isotope fractionations might influence the applicability of stable isotope analysis for quality and authenticity assessment of fruit brandies, the collected distillate fractions were recombined to cuts, as is common practice in commercial fruit brandy production. Taking into consideration the limits of repeatability of the method, it could be demonstrated that the isotope fractionations observed do not impair the applicability of stable isotope analysis of the carbon and hydrogen isotopes of ethanol for the authenticity assessment of cherry brandies if the cuts are placed in accordance with common distillers' practice.

Alcoholic Beverages↗
PubMed2006

Combined enzymatic and high-pressure processing affect cell wall polysaccharides in berries.

The effect of high-pressure processing (HPP) on cell wall polysaccharides in berries was investigated. HPP decreased the degree of methyl esterification (DM), probably by activation of pectin methyl esterase (PME), and improved the extractability of pectins. When commercial enzyme mixtures were added to mashed berries, a synergistic effect was observed between treatment with commercial enzymes and HPP. Compared to treatment at atmospheric pressure, pectic polysaccharides were degraded to a larger extent when HPP was used. In contrast, hemicelluloses were hardly affected by the added enzymes when HPP was included, although they were degraded during similar treatment at atmospheric pressure. Additionally, the activity of rhamnose-releasing enzymes present in minor quantities might be enhanced after HPP, resulting in a decrease of rhamnose in the polymeric cell wall material. These results exploring the effect of HPP at representative conditions clearly point out the potential of HPP for polysaccharide modification.

Acetylation↗
PubMed2006

Stability of barley and malt lipid transfer protein 1 (LTP1) toward heating and reducing agents: relationships with the brewing process.

Barley lipid transfer protein (LTP1) is a heat-stable and protease-resistant albumin that concentrates in beer, where it participates in the formation and stability of beer foam. Whereas the barley LTP1 does not display any foaming properties, the corresponding beer protein is surface-active. Such an improvement is related to glycation by Maillard reactions on malting, acylation on mashing, and structural unfolding on brewing. The structural stability of purified barley and glycated malt LTP1 toward heating has been analyzed. Whatever the modification, lipid adduction or glycation, barley LTP1s are highly stable proteins that resisted temperatures up to 100 degrees C. Unfolding of LTP1 occurred only when heating was conducted in the presence of a reducing agent. In the presence of sodium sulfite, the lipid-adducted barley and malt LTP1 displayed higher heat stability than the nonadducted protein. Glycation had no or weak effect on heat-induced unfolding. Finally, it was shown that unfolding occurred on wort boiling before fermentation and that the reducing conditions are provided by malt extract.

Beer↗
PubMed1999

Release of deuterated nonenal during beer aging from labeled precursors synthesized in the boiling kettle.

The use of labeled nonenal enabled the demonstration that the appearance of the cardboard flavor in finished beer comes from lipid auto-oxidation during wort boiling and not from lipoxygenasic activity during mashing. Free trans-2-nonenal produced by linoleic acid auto-oxidation in the kettle disappears, owing to retention by wort amino acids and proteins. This binding linkage protects trans-2-nonenal from yeast reduction but is reversible, allowing release of the compound at lower pH during aging. Labeled trans-2-nonenal is detected after aging when deuterated precursors form in the boiling kettle. The amount of alkenal released correlates with the concentration of reversible associations in the pitching wort. This work brings new illumination to the formation of trans-2-nonenal and overturns many previous hypotheses. It also explains why a reduction in the beer pH intensifies the cardboard flavor.

Aldehydes↗
PubMed1999

Expression of fungal thermotolerant endo-1,4-beta-glucanase in transgenic barley seeds during germination.

The malting quality of two barley cultivars, Kymppi and Golden Promise, was modified to better meet the requirements of the brewing process. The egl1 gene, coding for fungal thermotolerant endo-1,4-beta-glucanase (EGI, cellulase), was transferred to the cultivars using particle bombardment, and transgenic plants were regenerated on bialaphos selection. Integration of the egl1 gene was confirmed by Southern blot hybridization. The transgenic seeds were screened for the expression of the heterologous EGI. Under the high-pI alpha-amylase promoter, the egl1 gene was expressed during germination. The heterologous enzyme was thermotolerant at 65 degrees C for 2 h, thus being suitable for mashing conditions. The amount of heterologous EGI produced by the seeds (ca. 0.025% of soluble seed protein), has been shown to be sufficient to reduce wort viscosity by decreasing the soluble beta-glucan content. A decrease in the soluble beta-glucan content in the wort improves the filtration rate of beer.

Blotting, Southern↗
PubMed2002

Effects of scavenging on diet selection and performance of Rhode Island red and Fayoumi breeds of chicken offered a choice of energy and protein feeds.

An experiment was conducted to evaluate the effect of scavenging on diet selection and the comparative performance of Rhode Island Red and Fayoumi breeds of chicken. The breeds were compared under three feeding/management regimes: (1) a choice of ground noug cake and cracked maize, with confinement; (2) a choice of ground noug cake and cracked maize, but being allowed to scavenge for about 6 h; and (3) a commercial layer mash, with confinement. Scavenging did not change the pattern of diet selection. About 90% of the daily intake of both the confined and scavenging birds under choice feeding comprised maize. The efficiency of feed utilization and nutrient intake were also similar in these groups. The Fayoumi were more efficient in terms of feed conversion, although the egg production performance of the two breeds did not vary significantly. Scavenging hens had a significantly lower egg production than the birds under confinement offered a choice of feeds. Scavenging significantly increased the mortality in both breeds. In conclusion, scavenging affected the performance adversely and, despite the provision of a choice of feed providing a source of energy and protein, both the scavenging and confined birds on choice feeding failed to eat sufficient to meet their protein requirements.

Animal Feed↗
PubMed2004

Use of Zymomonas mobilis and Saccharomyces cerevisiae mixed with Kluyveromyces fragilis for improved ethanol production from Jerusalem artichoke tubers.

Jerusalem artichoke mashed tubers were fermented using single yeasts and a bacterium as well as mixed culture of microorganisms. Kluyveromyces fragilis, a yeast with an active inulinase, was used together with either a commercial distillery yeast, Saccharomyces cerevisiae, or the bacterium Zymomonas mobilis. After batch fermentation the best ethanol concentration of 0.48 g g(-1) for the mixed population and 0.46 g g(-1) for the single population can be obtained. The theoretical yield of the mixed cultures was 2-12% higher than for the single microorganism.

Bioreactors↗
PubMed1998

The interaction of diet and stress in rats: high-energy food and sucrose treatment.

Exposure to inescapable shock typically reduces eating and body weight in rats. The present study examined the modulation of stress effects by prestress diet and poststress sugar availability. Maintenance on a high-fat, high-energy food attenuated stress-induced weight loss and anorexia and increased high-energy food selection when a low-energy wet mash was the only alternative. Access to sugar after stress also reduced short-term weight loss; among rats maintained on high-energy food, body weight was spared absolutely. The dependence of stress effects on pre- and poststress diet alternatives may speak to individual differences in the stress-eating relationship in humans. More generally, these results support a conceptualization of stress in terms of metabolic challenge and the integrated reorganization of energy regulatory processes.

Animal Nutritional Physiological Phenomena↗