Effect of ethanol concentration and application period of soyabean meal on the kinetics of ruminal digestion

Effects of ethanol concentration and duration of treatment of soyabean meal on nitrogen (N) solubility, the kinetics of in situ dry matter, and N disappearance in the rumen were determined. Ground (1 mm) soyabean meal was soaked in 30%, 50%, 70% or 90% ethanol for 12, 24, 36 or 48 h. Seventy percent ethanol solutions applied to soyabean meal for 36 h or less lowered N solubility and the estimated rate of N disappearance, and increased the estimated rumen undegradable protein in soyabean meal compared to untreated soyabean meal. The lowest estimated rate of N disappearance and the highest increase in calculated rumen undegradable protein were observed when soyabean meal was treated with 70% ethanol for 36 h. Extending the application period of 70% ethanol to soyabean meal for more than 36 h removed the beneficial effect that the treatment had on N kinetics in the rumen and was considered excessive. Regardless of the application period, the 30%, 50%, and 90% ethanol solutions applied to soyabean meal did not lower the estimated rate of N disappearance in soyabean meal. However, extending the treatment time of soyabean meal with 90% ethanol for more than 24 h lowered the estimated rate of N disappearance compared to the shorter treatment times and the data suggested that 90% ethanol solutions applied to soyabean meal required longer application periods than 70% ethanol solutions to be effective. It was concluded that treatment of soyabean meal with 70% ethanol for 36 h had the greatest potential to increase rumen undegradable protein.     

进口转基因抗草甘膦油菜籽和大豆中CP4—EPSPS基因的检测比较研究

由于国际上对转基因产品的安全性存在的较大争议,因此对没有加施标签予以声明的情况下大量进入中国市场的转基因产品进行检测就显得十分迫切且意义重大。本研究以进口转基因抗草甘膦油菜籽和大豆为材料,通过比较分析外源的CP4－EPSPS基因的核苷酸序列和表达的氨基酸序列,设计出两对不同的引物,采用PCR方法分别对转基因油菜籽和大豆中外源的CP4－EPSPS基因进行了检测。     

A spin label study of the lipid boundary layer of mitochondrial NADH-ubiquinone oxidoreductase

Mitochondrial NADH-ubiquinone oxidoreductase (Complex I) is a lipoprotein enzyme containing phosphatidylcholine (PC), phosphatidylethanolamine (PE) and cardiolipin. Enzyme preparations containing endogenous cardiolipin and a range of either soyabean PC or dimyristoylphosphatidylcholine (DMPC) concentrations have been made. Using a spin-labelled fatty acid, two probe environments differing in mobility have been shown to be present. The fatty acid probe has a relative binding constant (or partition coefficient between lipid and protein) of unity. The boundary layer or lipid annulus reported by the probe has a value of approx. 300 lipid molecules per molecule of enzyme FMN in preparations containing soyabean PC, or DMPC above the phase transition temperature of the latter. In soyabean PC-replaced enzyme the apparent size of the boundary layer is independent of temperature between 30 degrees C and 14 degrees C but shows a modest increase to about 400 lipid molecules per molecule of FMN between 14 degrees C and 2 degrees C. Complex I replaced with high concentrations of DMPC gives non-linear Arrhenius plots of NADH-ubiquinone oxidoreductase activity. The results of the ESR experiments show that both boundary layer and bulk lipid must be motionally restricted for this to occur. Thus, the change in activity is probably not caused by an effect exerted directly on the catalytic activity of the enzyme but is more likely due to restriction of free diffusion of ubiquinone to its site of reduction.     

Neurotensin-like immunoreactivity generated by pepsin from human plasma and gastric tissue

The present study demonstrates precursors of neurotensin-like immunoreactivity (NTLI) endogenous to human gastric tissue and plasma, and the existence of a gastric NTLI-generating enzyme system. The molecular size of the NTLI-precursors in plasma and gastric tissue were estimated by gel permeation chromatography to be ca 50,000-60,000 and 60,000-70,000 Da, respectively. The neurotensin-like peptide generated from the precursor was detected with a carboxyl-terminally directed antiserum but did not cross-react with an amino-terminally directed antiserum. A neurotensin-like peptide isolated from pepsin-treated human plasma was characterized by mass spectrometry and its amino acid sequence determined. This novel nonapeptide, referred to as kinetensin, failed to affect pentagastrin-stimulated acid secretion or blood pressure in the rat. Sequence homologies between neurotensin, kinetensin and proteins of the serum albumin family suggest a common evolutionary origin and raise questions regarding albumin-like proteins as precursors of regulatory peptides.     

The kinetics of chemical reaction of activated nitrosamine with organic substrates.

The chemical reaction of tosylated N-nitrosobis (2-hydroxylpropyl) amine with aniline, 1-naphthol, pyridine, and hydroxylamine follows kinetics which can be described in a mixed order rate equation. This nitrosamine, considered an activated nitrosamine with a potential for alkylating a substrate, was seen to alkylate aniline, pyridine, 1-naphthol, and hydroxylamine. The presence of salts in the solution as well as an increase in solution pH increased the rate of reaction of the nitrosamine with substrates. The nitrosamine reacts quickly with water but will not react with solvents such as acetonitrile, dimethylsulfoxide, and methylene chloride.     

Dietary modification of lead toxicity: effects on fatty acid and eicosanoid metabolism in chicks

1. The influence of nutrition and chronic lead (Pb) administration on the level of eicosanoids and their precursors was evaluated in serum and liver from chicks fed defined diets. 2. Dietary Pb consistently increased arachidonic acid, the arachidonate/linoleate ratio, and hepatic non-protein sulfhydryl concentration. 3. Hepatic microsomal fatty acid elongation activity was decreased by Pb. 4. Cottonseed oil, calcium and methionine interacted with Pb, affecting liver and serum fatty acid profiles. 5. Tissue levels of leukotriene C4 rose significantly with Pb or correlated with Pb-compromised body weight in only one experiment. Serum prostaglandins E2 and F2 alpha were unchanged by treatments. 6. The results suggest that an increase in precursors is not consistently reflected by tissue levels of eicosanoids.     

Characterization of volatile compounds produced by Rhizopus strains grown on agro-industrial solid wastes

Four edible Rhizopus strains were cultivated on eight combinations of solid agro-industrial wastes (cassava bagasse, apple pomace), soyabean, amaranth grain and soyabean oil. Significant differences in growth were observed among strains on the different media studied. The medium containing cassava bagasse with soyabean (5:5 w/w) gave the highest CO₂ production, while Rhizopus oryzae ATCC 34612 was the best producer of volatiles. The aromas of the cultures were light and rather pleasant. The amaranth medium with mineral salts solution produced the highest amount of volatile compounds (VC), demonstrating that the aroma of fermented solid substrates can be improved. The VC production was very rapid, attaining, in most of the cases, its maximum around the first day of culture. These maxima markedly varied according to the medium used.     

Isoelectric precipitation of soy protein: I. Factors affecting particle size distribution

The particle size distribution (PSD) of soyabean protein aggregates obtained by acid precipitation in a continuous stirred reactor was measured with a Coulter Counter model TA II. Effects of changes in the operating variables pH, concentration, mean residence time, power input, and ionic strength were investigated. Power input affects both growth and breakup while pH and ionic strength seem to exert their influence through solubility and aggregate strength. Changes in zeta-potential can explain most of these observations.     

Incorporation of [32P]orthophosphate into brain-slice phospholipids and their precursors. Effects of electrical stimulation

1. The incorporation of [(32)P]phosphate into phospholipids was measured in slices cut from the pial surface of guinea-pig cerebral cortex; incorporation into the phosphorus of some water-soluble precursors of phospholipid was measured under similar conditions. 2. Slices subjected to overall electrical stimulation at a frequency of 5pulses/sec. differed from control slices in their pattern of phospholipid labelling. After 1hr. of stimulation, incorporation of [(32)P]phosphate into phosphatidylcholine, ethanolamine phospholipid and cardiolipin was respectively 54, 55 and 58% of the control value, and that into phosphatidylinositol was 186% of control. Phosphatidic acid labelling tended to increase with electrical stimulation, but the statistical significance of this change was marginal. Labelling of phosphatidylglycerol and di- and tri-phosphoinositides was not affected significantly by electrical stimulation. 3. Electrical stimulation of the tissue altered the specific radioactivities of water-soluble precursors of phospholipid. 4. The turnover rates of the phosphate groups of phospholipids were estimated approximately from the specific radioactivities of phospholipids and their precursors. Phosphatidylinositol (and its lipid-soluble precursors) showed the largest change in turnover rate in response to electrical stimulation of the tissue; the turnover rates of other lipids were also affected. Changes in the specific radioactivity of phospholipids did not correspond to changes in turnover in these experiments.     

Chemical composition of different sorts of soybeans and changes under the influence of weather and soil-climatic conditions]

The biochemical composition (the content of protein and lipids, the content of essential amino acids in the protein complex, the fatty acid composition of lipids) of seeds of eighteen soyabean varieties cultivated in different soil-climatic zones under various weather conditions was examined. Changes in the biochemical parameters were primarily associated with the genotype of the variety. The effect of weather, soil and climatic conditions was less marked. With an increase of the protein content the concentration of lysine and methionine in proteins tended to decrease. As long as the oil content in seeds grew, the concentration of oleic acid increased and that of linoleic and linolenic acids decreased.     

Antibacterial activity of Ocimum sanctum L. fixed oil

Ocimum sanctum fixed oil showed good antibacterial activity against Staphylococcus aureus, Bacillus pumilus and Pseudomonas aeruginosa, where S. aureus was the most sensitive organism. Sesame and soyabean oils also showed moderate activity against S. aureus. Higher content of linolenic acid in O. sanctum fixed oil could contribute towards its antibacterial activity. The antibacterial activity combined with anti-inflammatory and analgesic activities of the oil, could make it useful in inflammatory disorder resulting from staphylococcal infection.     

Molecular modeling of dissociative and non-dissociative chemisorption of nitrosamine on close-ended and open-ended pristine and Stone-Wales defective (5,5) armchair single-walled carbon nanotubes

The nitrosamine adsorbed on close-ended and open-ended pristine and Stone-Wales defective (5,5) armchair single-walled carbon nanotubes (SWCNTs) was studied using the B3LYP/6-31G(d) method. Structure optimization of all possible adsorption configurations based on the combination of two nitrosamine (amino- and imino-) isomers and four types of nanotubes was carried out. The most stable configuration for the nitrosamine adsorbed on the (5,5) armchair SWCNTs was found to be dissociative chemisorption. The adsorption energies of the most stable structures of the adsorption complexes of close-ended and open-ended pristine SWCNTs with the imino isomer of nitrosamine were −127.15 and −137.14 kcal mol⁻¹, respectively.     

Free sterols and glycolipids in the aleurone tissue of germinating wheat

The action of gibberellic acid on wheat aleurone tissue led to reduced levels of free sterols and glycolipids compared with control tissue. Radio-labelled precursors were not incorporated into sterols or glycolipids, although mevalonate and glycerol were incorporated into polyisoprenoid hydrocarbon and phospholipid respectively. It is concluded that sterols and glycolipids are not synthesized in the tissue during germination; this is in contrast to earlier reports of the active, gibberellin-regulated metabolism of phospholipids.     

Aspects of adipose-tissue metabolism in foetal lambs.

1. The mean volume of adipocytes, the rates of fatty acid and acylglycerol glycerol synthesis from various precursors (in vitro), the rates of oxidation of acetate and glucose (in vitro) and the activities of lipoprotein lipase and various lipogenic enzymes were determined for perirenal adipose tissue from foetal lambs during the last month of gestation. 2. The fall in the rate of growth of perirenal adipose tissue during the last month of gestation is associated with a diminished capacity for fatty acid synthesis and lipoprotein lipase activity, but no change in the rate of acylglycerol glycerol synthesis was observed. There was no fall in the activities of cytosolic acetyl-CoA synthetase or the NADP-linked dehydrogenases, suggesting that the decrease in the rate of fatty acid synthesis was due to an impairment at the level of acetyl-CoA carboxylase or fatty acid synthetase. 3. The rate of fatty acid synthesis from acetate was greater than that from glucose. The rate of fatty acid synthesis from glucose per adipocyte of foetal lambs was similar to that of young sheep. The characteristic metabolism of adipose tissue of the adult sheep is thus present in the foetus, despite the relatively large amounts of glucose in the foetal 'diet'.     

Evidence for the occurrence of an alkaline proteinase in kidney cortex lysosomes

A novel alkaline endoproteinase optimally active at pH 8.5 has been detected in highly pure preparations of buffalo kidney cortex lysosomes. The enzyme has been partially purified (90-fold) by solubilization with octylglucoside, acid precipitation and chromatography over DEAE sephacel and sepharose 6B. The alkaline proteinase, resistant to known inhibitors of lysosomal cathepsins is inhibited by soyabean trypsin inhibitor and phenyl methyl sulphonyl fluoride indicating that the enzyme is a serine proteinase.     

Pyrimidine biosynthesis and its regulation in the estrogen-stimulated chick oviduct.

Studies on the incorporation of radio-labeled precursors into orotic acid and the pyrimidine nucleotides of RNA have established the occurrence of the orotate pathway for the de novo biosynthesis of pyrimidines in the chick oviduct. Measurements of the rate of incorporation of precursors into orotic acid in minces of oviduct revealed the activity of the orotate pathway to be accelerated in response to estrogen-stimulated nucleic acid synthesis and tissue growth. These data indicate that extrahepatic tissues of avian species meet their requirements for pyrimidine nucleotides through de novo synthesis rather than depend upon the liver or other exogenous sources for a supply of preformed pyrimidines. An examination of the influence of pyrimidine and purine nucleosides on the incorporation of radio-labeled precursors into orotic acid yielded evidence that pyrimidine biosynthesis in the chick is quite sensitive to inhibition by both purines and pyrimidines; the data indicate the reaction catalyzed by carbamoylphosphate synthetase to be the site of inhibition in both cases.     

EFFECT OF PROLACTIN ON THE TADPOLE TAIL FIN. IV. EFFECT OF PROLACTIN ON THE METABOLIC FATE OF HYALURONIC ACID, COLLAGEN AND RNA WITH A SPECIAL REFERENCE TO CATABOLIC PROCESS

In the case of hyaluronic acid and collagen, treatment of tadpoles with bovine prolactin produced remarkable increase in the content of the two metabolites in the tail fin in accord with the stimulatory effect of the hormone on the incorporation of labeled precursors into hyaluronic acid and collagen. RNA content of the tissue was increased appreciably by the prolactin treatment, whereas the hormone had no effect on the incorporation of ¹⁴C-uridine into RNA. Prolactin had inhibitory effect on the activities of hyaluronidase and RNase of the tail fin. The experiments on the protein catabolism also revealed that prolactin inhibited the process of protein turn-over in the tissue.     

Microheterogeneity of rat submaxillary gland kallikrein k10, a member of the kallikrein family

A tissue-kallikrein-related proteinase present in rat submaxillary glands, which was previously called endopeptidase k, has been further characterized and compared with other members of the kallikrein family. The partial primary structure of this proteinase, now called kallikrein k10, is very similar to that of proteinase B [Kato, H., Nakanishi, E., Enjyoji, K., Hayashi, I., Oh-Ishi, S. & Iwanaga, S. (1987) J. Biochem. (Tokyo) 102, 1389-1404] and T-kininogenase [Xiong, W., Chen. L. M. & Chao, J. (1990) J. Biol. Chem. 265, 2822-2827], but no corresponding gene or mRNA has so far been found. Kallikrein k10 is microheterogeneous due to variable glycosylation of its N-terminal light chain and to variable processing at its kallikrein loop, as shown by endo-beta-N-acetylglucosaminidase F treatment, amino acid sequence analysis and mass spectrometry. The enzymatic properties of the two molecular varieties of kallikrein k10 towards synthetic fluorogenic substrates are not significantly different. Both cleave specifically after Arg residues, but, in contrast to true tissue kallikrein, may accommodate either polar or nonpolar residues at position P2. Kallikrein k10 also differs from tissue kallikrein by its sensitivity to soyabean trypsin inhibitor. Its biological function may therefore differ from that of tissue kallikrein, especially as it does not induce a transient decrease in blood pressure when injected in vivo.     

Reductive metabolism of N-nitrosodiphenylamine to the corresponding hydrazine derivative

The first evidence for the reductive metabolism of a noncyclic nitrosamine to the corresponding hydrazine derivative in vivo and in vitro is provided. Under anaerobic conditions, N-nitrosodiphenylamine was reduced by guinea pig liver 9000g supernatant to 1,1-diphenylhydrazine in the presence of 2-hydroxypyrimidine or to acetaldehyde diphenylhydrazone in the presence of acetaldehyde. These metabolites were identified unequivocally by comparative study with authentic samples. In addition, the study shows that such reductive reactions of the nitrosamine can be catalyzed by guinea pig and rabbit liver aldehyde oxidase in the presence of its electron donors. When the nitrosamine was given orally to acetaldehyde-treated guinea pigs, a metabolite was detected from plasma and identified as acetaldehyde diphenylhydrazone by comparison with the authentic sample.     

Evidence for similar structural changes on binding of platinum anti-tumor agents to DNA and nucleosomes

D2 dopamine receptors have been solubilised from bovine caudate nucleus using cholate/sodium chloride in the presence of soyabean phospholipid. Reconstitution of the receptors into soyabean phospholipid vesicles has been achieved by dialysis to remove detergent and salt. The receptors are truly reconstituted as judged by sedimentation, electron microscopy, heat stability and analysis on sucrose density gradients. The ligand-binding properties of the reconstituted receptors resemble those of the solubilised preparation.