Resistant starch reduces colonic and urinary p-cresol in rats fed a tyrosine-supplemented diet,whereas konjac mannan does not

The effect of resistant starch (RS) and konjac mannan (KM) to maintain and improve the large intestinal environment was compared. Wistar SPF rats were fed the following diets for 4 weeks: negative control diet (C diet), tyrosine-supplemented positive control diet (T diet), and luminacoid supplemented diets containing either high-molecular konjac mannan A (KMAT diet), low-molecular konjac mannan B (KMBT diet), high-amylose cornstarch (HAST diet), or heat-moisture-treated starch (HMTST diet). The luminacoid-fed group had an increased content of short-chain fatty acids in the cecum. HAS caused a significant decrease in p-cresol content in the cecum, whereas KM did not. Urinary p-cresol was reduced in the HAST group compared with the T group, but not the KM fed groups. Deterioration in the large intestinal environment was only improved completely in the HAST and HMTST groups, suggesting that RS is considerably more effective than KM in maintaining the large intestinal environment.     

Possible role of glutamate, aspartate, glutamine, GABA or taurine on cadmium toxicity on the hypothalamic pituitary axis activity in adult male rats

This work was designed to evaluate the possible changes in glutamate, aspartate, glutamine, GABA and taurine within various hypothalamic areas the striatum and prefrontal cortex after oral cadmium exposure in adult male rats, and if these changes are related to pituitary hormone secretion. The contents of glutamine, glutamate, aspartate, GABA and taurine in the median eminence, anterior, mediobasal and posterior hypothalamus, and in prefrontal cortex in adult male rats exposed to 272.7 mol l^–1 of cadmium chloride (CdCl₂) in the drinking water for one month. Cadmium diminished the content of glutamine, glutamate and aspartate in anterior hypothalamus as compared to the values found in the untreated group. Besides, there is a decrease in the content of glutamate, aspartate and taurine in the prefrontal cortex. The amino acids studied did not change in median eminence, mediobasal and posterior hypothalamus or the striatum by cadmium treatment. Plasma prolactin and LH levels decreased in rats exposed to the metal. These results suggest that (1) cadmium differentially affects amino acid content within the brain region studied and (2) the inhibitory effect of cadmium on prolactin and LH secretion may be partially explained by a decrease in the content of both glutamate and aspartate in anterior hypothalamus, but not through changes in GABA and taurine.     

不依赖连接反应的高通量克隆方法

以脱脂菜籽粕酶解的复合氨基酸为主要原料,螯合率为考察指标,采用L16(4 5)正交试验设计,考察了pH值、温度、时间和配位比对螯合率的影响.结果显示,影响因素的高低顺序为:配位比＞pH值＞时间＞温度.结果表明,复合氨基酸与铜螯合的主要影响因素为pH值和配位比,且配位比的影响达到极显著水平.最佳工艺条件为:时间50 min,温度50℃,配位比2∶1,pH为9,此条件下的复合氨基酸螯合铜的螯合率为94.59％,氨基酸含量为30.2％.     

An isocaloric moderately high-fat diet extends lifespan in male rats and Drosophila

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支链氨基酸在运动中的作用研究进展

支链氨基酸(BCAAs)由亮氨酸、异亮氨酸和缬氨酸组成,是人体重要的必需氨基酸,参与人体多种生理活动。本文在大量查阅相关文献的基础上,就支链氨基酸在运动中的作用进行述评。支链氨基酸在运动中能够为机体提供能量,通过参与糖代谢过程而维持糖原含量;可调节蛋白质的代谢过程,抑制蛋白质降解并促进合成代谢;抑制自由基和乳酸的产生并加快其廓清速率,保护细胞膜与线粒体的生物功能,减缓抑制性神经递质的生成与集聚。在对抗运动性疲劳、提高运动能力和削弱延迟性肌肉酸痛方面有巨大影响。但关于长期补充支链氨基酸的毒副作用、不良效应等问题,仍有待进一步研究与探索。     

Growth rate of male germfree Wistar rats fed ad libitum or restricted natural ingredient diet

Germfree life and feed restriction initiated at an early age are known to extend life span. We have examined growth rate and life expectancy in germfree (GF) and conventional (CV) male Lobund Wistar rats, fullfed (F) or restricted (R) to 12 grams/day of natural ingredient diet L-485. GF-F and CV-F rats show comparable growth rates during the first 6 months of life. Thereafter, the GF-F rat falls behind, with its body weight stabilizing at 85% of the CV rat's weight at 2 years of age (510 g vs 435 g). In contrast, GF-R rats become slightly, but significantly, heavier than CV-R rats after an initial 6 months of comparable growth. At 2 years of age GF-R rats weigh 12% more than the CV-R rats (340 g vs 300 g). Physiological parameters were examined in each treatment group in animals that had to be sacrificed because of contamination. These gnotobiotic (GN) rats (see text) and their CV counterparts were grouped as adult (7 to 11 months) and old (18 to 28 months) rats. The most significant findings were: GN-F rats have smaller hearts than CV-F rats, both on an absolute and relative basis; restriction did not affect absolute testes size but elevated serum testosterone levels; serum T4 was reduced by restriction only in CV rats, and declined with age in all groups; and serum T3 was higher in adult GN-F and GN-R rats, but fell to CV levels in old age.(ABSTRACT TRUNCATED AT 250 WORDS)     

Reconstitution of Neutral Amino Acid Transport Systems from Ehrlich Ascites Tumor Cells

Amino acid transport systems for alanine and leucine were reconstituted into artificial lipid vesicles. Purified plasma membrane vesicles from Ehrlich ascites cells were dissolved in 2% sodium cholate, 1mM dithiothreitol, and 0.5 mM EDTA a mixture that solubilized approximately 50% of the membrane protein. This solubilized protein fraction was further purified by a combination of ammonium sulfate precipitations, gel filtration, and DEAE-cellulose chromatography. A fraction containing approximately 15 Coomassie blue-staining bands on sodium dodecyl sulfate gels was obtained. This material was reconstituted into liposomes, and preliminary results demonstrated transport of alanine and leucine dependent on a sodium gradient. In addition, an electrogenic gradient mediated by valino-mycin-induced potassium diffusion seemed to stimulate alanine uptake further.     

The Crystal Structure of L-Leucine Dehydrogenase from Pseudomonas aeruginosa

Leucine dehydrogenase (LDH, EC 1.4.1.9) catalyzes the reversible deamination of branched-chain L-amino acids to their corresponding keto acids using NAD⁺ as a cofactor. LDH generally adopts an octameric structure with D4 symmetry, generating a molecular mass of approximately 400 kDa. Here, the crystal structure of the LDH from Pseudomonas aeruginosa (Pa-LDH) was determined at 2.5 _Å resolution. Interestingly, the crystal structure shows that the enzyme exists as a dimer with C2 symmetry in a crystal lattice. The dimeric structure was also observed in solution using multiangle light scattering coupled with size-exclusion chromatography. The enzyme assay revealed that the specific activity was maximal at 60°C and pH 8.5. The kinetic parameters for three different amino acid and the cofactor (NAD⁺) were determined. The crystal structure represents that the subunit has more compact structure than homologs’ structure. In addition, the crystal structure along with sequence alignments indicates a set of non-conserved arginine residues which are important in stability. Subsequent mutation analysis for those residues revealed that the enzyme activity reduced to one third of the wild type. These results provide structural and biochemical insights for its future studies on its application for industrial purposes.     

Reconstitution of Neutral Amino Acid Transport System from Ehrlich Ascites Tumor Cells

Amino acid transport systems for alanine and leucine have been reconstituted into artificial lipid vesicles. Purified plasma membrane vesicles from Ehrlich ascites cells were dissolved in 2% sodium cholate, 1 mM dithiothreitol, 0.5 mM EDTA, a mixture which solubilized approximately 50% of the membrane protein. This solubilized protein fraction was further purified by a combination of ammonium sulfate precipitations, gel filtration, and DEAE-cellulose chromatography. A fraction containing approximately 15 Coomassie blue staining bands on sodium dodecyl sulfate gels was obtained. This material was reconstituted into liposomes, and preliminary results demonstrated transport of alanine and leucine dependent on a sodium gradient. In addition, an electrogenic gradient mediated by valinomycin-induced potassium diffusion seemed to stimulate alanine uptake further.     

Stimulatory effect of a maize diet on sexual behaviour of male rats

Feeding rats for 4 days with a diet of maize, a staple which is deficient in tryptophan (TP), caused a depletion of brain 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) without affectin noradrenaline and dopamine content.Addition of TP (2 g/kg of diet) to the TP deficient diet resulted in increased brain 5-HT and 5-HIAA content.Isolated male rats consuming the maize diet, when brought together, showed markedly increased mounting activity, while rats on the maize diet supplemented with TP did not display such a behaviour.These data provide further evidence for the role of the 5-HT system in regulating the sexual behaviour of male rats.     

Dietary CP and amino acid restriction has a different impact on the dynamics of protein,amino acid and fat deposition in entire male,castrated and female pigs

Breeding efforts over the last decades altered markedly empty body (EB) composition of pigs. This study aimed to re-evaluate the dynamics of changes in the composition and deposition rate of fat, protein and amino acids (AA) in the EB from birth to 140 kg BW depending on the dietary CP and AA supply in a current pig genotype. In the experiment 66 entire male, 58 castrated and 66 female Swiss Large White pigs were used. From 20 kg BW onwards, they had either ad libitum access to a control (C) diet or a diet (LP) compared to diet C only 80% of CP, lysine, methione+cystine, threonine and tryptophan. The EB composition was determined at birth on eight boars and eight females, at 10 and 20 kg BW on two boars, two castrates and two females, and at 20 kg intervals from 40 to 140 kg BW, on four pigs per gender and dietary treatment. Each EB fraction was weighed and analysed for protein, fat and AA profile. The AA-to-lysine ratio was calculated and the different chemical component contents were fitted to allometric regressions. Overall, C-boars had the greatest EB protein and AA content and deposition rates, and lowest fat content and deposition rates. At the beginning of the grower period, LP-castrates and females displayed the lowest protein and AA and the highest fat deposition rates. However, compared with their counterparts in the C-group, in LP-castrates and females protein and AA deposition rates were greater above 64 and 40 kg EB weight, respectively, whereas fat deposition rates was lower above 80 kg EB weight. Thus, there seems a great potential to optimise protein and AA efficiency especially in the finisher period in castrates and females. Important individual variations were found in the essential AA-to-lysine ratio of the EB. Phenylalanine and threonine-to-lysine ratios decreased with increasing EB weight. Valine- and threonine-to-lysine ratios in C-castrates and C-females were 5% and 4% greater than recently reported by the National Research Council (NRC) whereas cysteine-, methionine- and tyrosine-to-lysine ratios were lower by 34%, 25% and 10%, respectively. The clear differences found between the EB AA-to-lysine ratios in the present study and the NRC might partly be explained by the genotype and the temporal changes in the relative weight of each EB fraction or changes in the AA profile. Nevertheless, these findings on changes in the essential AA profile of tissue protein warrant further studies.     

Biological and biochemical characteristics for quality control of Harmonia axyridis (Pallas) (Coleoptera, Coccinellidae) reared on a liver-based diet

Biological and biochemical parameters of a flightless strain of Harmonia axyridis, fed on a pork liver-based artificial diet and on Ephestia kuehniella eggs as controls, were compared. The diet-grown larvae showed a significantly longer developmental time and a lower adult emergence rate compared to control larvae. The weights of the newly emerged adults were significantly higher for adults fed E. kuehniella eggs during their larval stages than fed the artificial diet. In contrast, larval food source had no effect on the duration of the pre-oviposition period or adult longevity. For adults fed on E. kuehniella eggs as larvae, a significantly longer pre-oviposition period, lower daily weight gain and fecundity were found for the diet-fed females compared to those fed on E. kuehniella eggs throughout the life span. The adult food source had no significant effect on longevity and fertility. Lower amino acid and fatty acid contents (in particular C16:1 and C18:3n-3) were found for the prepupae and newly emerged females obtained from diet-reared larvae compared to controls. Deficiencies in fatty acids C16:1 and C18:3n-3 were also observed in females obtained from E. kuehniella egg-reared larvae and fed on diet from adult emergence. The analyses of the foods showed deficiencies in artificial diet, especially for some amino and fatty acids. The results suggest a non-optimal composition of the artificial diet and some possibilities for its improvement. However, this polyphagous predator could be reared from first instar larvae to fully reproductive adults on a pork liver-based artificial diet.     

Annual changes in the copulatory behavior of male rats maintained under constant laboratory conditions

ABSTRACT

The objective of the present study was to evaluate the sexual behavior of male rats kept under constant laboratory conditions for one entire year. A total of 213 sexually-inexperienced, male Wistar rats were maintained in controlled environmental conditions from birth. Depending the month in which they reached the age 3-month-old, the male rats were divided into 12 groups, one for each month of the year, and their sexual behavior was evaluated. Records of their sexual behavior were made from 09:00 to 11:00 hrs am. The following parameters were recorded: mount (latency and number), intromission (latency and number), ejaculation latency, and intromission rate. During the months of March, June, July and September, the rats showed lower mount and intromission latencies than in January, February, April, May and October-to-December. Similarly, in March, June, July and August they had higher copulatory efficiency than in January, February, April and December. Results suggest that male rats exposed to controlled environmental conditions could have endogenous mechanisms that regulate sexual behavior but are independent of seasonal environmental signals. The annual variability in the sexual behavior of male rats maintained under constant laboratory conditions should be considered when planning research and experiments.     

Widespread neuronal expression of branched-chain aminotransferase in the CNS: implications for leucine/glutamate metabolism and for signaling by amino acids

Transamination of the branched-chain amino acids produces glutamate and branched-chain alpha-ketoacids. The reaction is catalyzed by branched-chain aminotransferase (BCAT), of which there are cytosolic and mitochondrial isoforms (BCATc and BCATm). BCATc accounts for 70% of brain BCAT activity, and contributes at least 30% of the nitrogen required for glutamate synthesis. In previous work, we showed that BCATc is present in the processes of glutamatergic neurons and in cell bodies of GABAergic neurons in hippocampus and cerebellum. Here we show that this metabolic enzyme is expressed throughout the brain and spinal cord, with distinct differences in regional and intracellular patterns of expression. In the cerebral cortex, BCATc is present in GABAergic interneurons and in pyramidal cell axons and proximal dendrites. Axonal labeling for BCATc continues into the corpus callosum and internal capsule. BCATc is expressed by GABAergic neurons in the basal ganglia and by glutamatergic neurons in the hypothalamus, midbrain, brainstem, and dorsal root ganglia. BCATc is also expressed in hypothalamic peptidergic neurons, brainstem serotoninergic neurons, and spinal cord motor neurons. The results indicate that BCATc accumulates in neuronal cell bodies in some regions, while elsewhere it is exported to axons and nerve terminals. The enzyme is in a position to influence pools of glutamate in a variety of neuronal types. BCATc may also provide neurons with sensitivity to nutrient-derived BCAAs, which may be important in regions that control feeding behavior, such as the arcuate nucleus of the hypothalamus, where neurons express high levels of BCATc.     

Site-specific experiments on folding/unfolding of Jun coiled coils: thermodynamic and kinetic parameters from spin inversion transfer nuclear magnetic resonance at leucine-18

The 32-residue leucine zipper subsequence, called here Jun-lz, associates in benign media to form a parallel two-stranded coiled coil. Studies are reported of its thermal unfolding/folding transition by circular dichroism (CD) on samples of natural isotopic abundance and by both equilibrium and spin inversion transfer (SIT) nuclear magnetic resonance (NMR) on samples labeled at the leucine-18 alpha-carbon with 99% 13C. The data cover a wide range of temperature and concentration, and show that Jun-lz unfolds below room temperature, being far less stable than some other leucine zippers such as GCN4. 13C-NMR shows two well-separated resonances. We ascribe the upfield one to 13C spins on unfolded single chains and the downfield one to 13C spins on coiled-coil dimers. Their relative intensities provide a measure of the unfolding equilibrium constant. In SIT NMR, the recovery of the equilibrium magnetization after one resonance is inverted is modulated in part by the unfolding and folding rate constants, which are accessible from the data. Global Bayesian analysis of the equilibrium and SIT NMR data provide values for the standard enthalpy, entropy, and heat capacity of unfolding, and show the latter to be unusually large. The CD results are compatible with the NMR findings. Global Bayesian analysis of the SIT NMR data yields the corresponding activation parameters for unfolding and folding. The results show that both reaction directions are activated processes. Activation for unfolding is entropy driven, enthalpy opposed. Activation for folding is strongly enthalpy opposed and somewhat entropy opposed, falsifying the idea that the barrier for folding is solely due to a purely entropic search for properly registered partners. The activation heat capacity is much larger for folding, so almost the entire overall change is due to the folding direction. This latter finding, if it applies to GCN4 leucine zippers, clears up an extant apparent disagreement between folding rate constants for GCN4 as determined by chevron analysis and NMR in differing temperature regimes.     

The accumulation and metabolism of amino acids by Biomphalaria glabrata, a freshwater pulmonate snail

Freshwater pulmonate snails (Biomphalaria glabrata), pre-treated under bacteriostatic conditions, were incubated in 10 ml of standard medium containing various U-¹⁴C-labelled amino acids at concentrations of 10 μM. Measurements of mass-specific accumulation rates (MSARs) based on HPLC and the accumulation of U-¹⁴C-labelled amino acids into snail tissues have shown unequivocally for the first time that freshwater snails achieved a net accumulation of all the amino acids tested, including aminoisobutyrate (AIB), aspartate, alanine and a mixture of 13 amino acids. There were no significant differences between the MSAR values determined by HPLC from those based on the use of radiolabelled amino acids, whereas MSAR values for control snails were negligible and significantly less. Incubation of snails in media containing radiolabelled aspartate and a mixture of amino acids showed that the accumulated amino acids were readily distributed through the snail’s tissues and then metabolized. The ecological and biochemical questions arising from the fact that freshwater snails are capable of net accumulation of exogenous amino acids at naturally occurring concentrations and subsequent metabolic conversion, contrary to widely held views, are addressed.