GABA and Dopamine Release from Different Brain Regions in Mice with Chronic Exposure to Organophosphate Methamidophos

Organophosphates such as methamidophos, usually used in the agricultural field, have harmful effects on humans. Exposures to insecticides has been associated with many disorders, including damage to the central and peripheral nervous system. Chronic exposure to organophosphates may lead to persistent neurological and neurobehavioral effects. This study was conducted to determine the effect of methamidophos on [(3)H]-dopamine (DA) and gamma aminobutyric acid (GABA) release from different brain regions after chronic exposure to it for 3, 6 or 9 months. After a six-month methamidophos treatment, the mice showed high susceptibility to convulsive seizures and a reduction in stimulated gamma aminobutyric acid release from the cerebral cortex and hippocampal slices, whereas stimulated (DA) release was slightly decreased from the striatum after three months of methamidophos exposure. The results indicate changes in gamma aminobutyric acid and dopamine neurotransmission, suggesting a specific neuronal damage.     

Polymorphisms in the canine glutamate transporter-1 gene: identification and variation among five dog breeds

Excitatory amino acid transporters (EAATs) are important for terminating glutamatergic neurotransmission and protect central nervous system (CNS) neurons from glutamatergic excitotoxicity. We selected these genes as targets that may relate to canine behavioral traits. After screening four EAAT genes (glutamate transporter-1; GLT-1, excitatory amino acid transporter 4; EAAT4, excitatory amino acid carrier; EAAC1, glutamate/aspartate transporter; GLAST) for single nucleotide polymorphisms (SNPs), we identified two silent SNPs (C129T and T471C) in the GLT-1 gene. We genotyped 193 dogs of 5 breeds and found significant variation among breeds in these two SNPs in GLT-1. The C129T polymorphism was not observed in Malteses and Miniature Schnauzers. These results suggest that polymorphisms in the GLT-1 gene may be useful markers for examining how the genetic background relates to the behavioral traits of dogs.     

Purification and comparison of corticosteroid-induced and intestinal isoenzymes of alkaline phosphatase in dogs

Corticosteroid-induced alkaline phosphatase (CALP) and intestinal alkaline phosphatase (IALP) from dogs were purified to homogeneity, as determined by polyacrylamide gel electrophoresis. Purification involved an un-interrupted system using DEAE-cellulose, concanavalin A-agarose, and monoclonal antibody affinity columns. The monoclonal antibody was prepared by use of IALP as the antigen. The 2 isoenzymes were compared, using molecular weight determinations, amino acid analyses, peptide mapping, N-terminal sequencing of the first 10 amino acids, carbohydrate analyses, and recognition by anti-IALP monoclonal antibody. The data indicated that canine IALP and CALP are identical with regard to recognition by monoclonal antibody and N-terminal amino acid sequence, nearly identical in amino acid content and peptide maps, but different in carbohydrate content. It was concluded that CALP is a product of the same gene as IALP and that differences in glycosyl transferase activities between liver and intestines or the presence of glycosidase activities in or around the intestinal mucosae result in the marked difference in carbohydrate content.     

Additive and synergistic pharmacologic inhibition of equine fibrinoligase (factor XIIIa*-like) biochemical activity.

A selected group of pharmaceutical compounds were evaluated for the ability to inhibit the biochemical activity of fibrinoligase (coagulation factor XIIIa*) in pooled equine plasma. Criteria for the pharmaceuticals selected were based on the mechanism of the transglutamination biochemical reaction mediated by coagulation factor XIIa*. These criteria were complemented by recognition of the molecular configuration and chemical composition of amino acid residue side chains involved in the process of covalent fibrin monomer polymerization (cross-linking, transglutamination) mediated by this enzyme. Each pharmaceutical was evaluated individually and in combination with other potential coagulation factor XIIIa* inhibitors in an effort to detect additive and synergistic phenomenon. In this context, pharmaceuticals with a carbonylamide (eg, cefuroxime, Girard's reagent-P, prolinamide) were applied in concert with compounds with a terminal amine (eg, D-arginine, L-lysine) functional group. In concept, this method theoretically served to competitively simulate glutamine and lysine amino acid residues within strands of fibrin monomer substrate involved in phase I (carbonylamide) and phase II (terminal amine) of the transglutamination reaction (covalent fibrin monomer cross-linking). Halogen-dinitro and ethylene compounds were also evaluated because of their reported ability to inactivate enzyme systems dependent on an intact sulfhydryl group located at their biochemically active site (eg, cystine amino acid residue). This group of pharmaceutical compounds failed to inhibit the biochemical activity mediated by coagulation factor XIIIa* in equine plasma.     

Evaluating the Technique of Using Nitrogen Retention as a Response Criterion for Amino Acid Studies in the Horse

Six Quarter Horse yearling fillies were used in a duplicated 3 × 3 Latin square experiment to evaluate the technique of using nitrogen retention as a response criterion for amino acid studies in the horse. Yearlings were paired by age and randomly assigned to one of three concentrates fed with medium-quality Coastal Bermudagrass hay. Diet A, a soybean meal–based concentrate, was amino acid sufficient. Diet B, a cottonseed hull–based concentrate containing 2% urea, was amino acid deficient. Horses fed diet C, identical to diet B, were orally dosed with synthetic essential amino acids and cysteine, immediately before eating, to match amino acid levels in diet A. Nitrogen retention was not different between diets A and B. There was a trend for horses fed diet C to have higher nitrogen retention, as a result of higher nitrogen intake. Nitrogen retained as a percent absorbed was lower (P < .05) for diet B than for diet A, whereas values for diet C were not different from diets A or B. Nitrogen retention was not an effective response criterion in this study. Differences in nitrogen retained as a percentage absorbed were observed between amino acid–sufficient and deficient diets. However, horses fed amino acid–deficient diets and dosed with synthetic amino acids likely require some modified dosage level to achieve the same or higher values as those for amino acid–sufficient diets.     

Comparison of amino acid digestibility using the ileal digesta from growing chickens and cannulated adult cockerels

Meat-and-bone meal (MBM), which had been heated (150 degrees C) for 0, 1.5, 3 or 5 h, was used along with an indigestible marker in four diets which were fed to young growing chickens and adult cockerels fitted with ileal cannulae. The ileal digesta from each group of birds were sampled and the apparent amino acid digestibilities of the four diets containing MBM were determined. The apparent digestibility values from growing chickens were higher (P less than 0.05) than those from cannulated cockerels. Differences in apparent digestibility of amino acids between diets as a result of heat treatment were consistent for all amino acids when comparing both techniques, with the exception of glutamic acid and arginine.     

Digestibility of amino acids in swine: Results and their practical applications. A review

Many studies have been carried out on the topic of amino acid digestibility in swine during the last two decades. The ileal analysis method should be the method of choice for determining amino acid digestibilities. This method is very sensitive for detecting differences in amino acid digestibilities, as these result from processing conditions or from inherent differences between samples of the same feedstuff. Amino acid digestibility values from the literature, determined with the ileal analysis method, are summarized and show large differences in amino acid digestibilities between feedstuffs and among different samples of the same feedstuff. At present it seems most appropriate to relate to amino acid digestibilities in terms of their apparent values. Studies in which the ¹⁵N isotope dilution technique was used to determine the endogenous amino acid losses have shown that the traditional methods for determining the endogenous amino acid losses should be re-evaluated. Recent studies have clearly shown improvements in diet formulation practices, especially when a good quality protein supplement is replaced by protein supplement(s) of lower quality combined with supplementary amino acids, and when diets are formulated on the basis of digestible rather than total supply of the limiting amino acid(s). In addition, in order to present improved values for amino acid requirements, these should be expressed as digestible rather than as total.     

绵羊催乳素受体基因外显子10的多态性分析

将催乳素受体(prolactin receptor, PRLR)基因作为绵羊高繁殖力的候选基因,对其外显子10设计2对引物, 采用PCR SSCP 技术检测其在常年发情的湖羊及季节性发情的中国美利奴羊、罗米丽羊和罗米丽×中国美利奴（新疆军垦型）中的单核苷酸多态性。结果表明,引物P1、P2 的扩增片段均有多态性。与已知序列相比,P1扩增片段的AB型在第53 bp处出现A→G突变、在81 bp处出现G→A突变, BB型在该片段第53 bp处发生A→G的突变;对于P2扩增片段, CC、DD、CE和EF型均在第89 bp处发生C→T的突变,导致氨基酸由脯氨酸变为亮氨酸(Pro→Leu),DD型还在146 bp处出现了C→G的突变,此突变导致氨基酸由丙氨酸变为甘氨酸(Ala→Gly);CE型在该片段第132 bp处发生G→A的突变,未导致氨基酸的改变;EF型还在第132 bp处和第167 bp处分别发生了G→A、C→T突变,第167 bp处的突变导致氨基酸由脯氨酸变为亮氨酸(Pro→Leu)。通过卡方独立性检验结果发现,4种绵羊在P1、P2引物扩增片段上各基因型的构成与品种间有极显著差异(P<0.01),说明PRLR基因对绵羊的繁殖性状有一定的影响。     

Dietary crude protein has minimal effect on the activity of selected enzymes of methionine catabolism in kittens fed diets near-limiting in methionine

Previous experiments have shown that increasing the dietary crude protein (CP) of cats does not increase urea cycle enzymes or alanine amino transferase as occurs in rats. Also when an essential amino acid (EAA) is limiting in a diet for growing kittens, the kittens do not exhibit an amino acid imbalance when other EAAs are added to the diet. To study the metabolic basis for these observations which are different from that found in omnivores and herbivores, the hypothesis that increased dietary CP decreases methionine catabolism, so more is spared for growth, was tested. Fifteen male kittens were randomly assigned to one of three dietary treatments. Each diet contained 2.5 g l-methionine/kg diet and 200, 300 or 500 g CP/kg diet. The livers and kidneys were removed and assayed for methionine transaminase (MTA), cystathionase (CASE) and cystathionine synthase (CS). Free amino acid concentrations were determined in liver, kidney and plasma. The 300 and 500 g CP/kg groups had significantly greater kidney weights and body weight gains than the 200 g CP/kg group. Hepatic MTA activity was lower in the 300 than the 200 or 500 g CP/kg groups (p < 0.05). Renal MTA and CASE activities were 35% and 50% greater, respectively, for the 500 g CP/kg group than for the 200 g CP/kg diet group (p < 0.05). Renal CS activities for the 300 and 500 g CP/kg groups were 29% (p > 0.05) and 38% (p < 0.05) greater, respectively, than the 200 g CP/kg group. Cyst(e)ine concentrations were lower in the livers of the 500 g CP/kg group than the 200 g CP/kg group (p < 0.05). Cystathionine was lower in plasma and kidney from the 500 g CP/kg diet group than from the 200 g CP/kg diet group (p < 0.05). It was concluded that the metabolic basis for the increased growth of kittens fed diets marginally limiting in methionine, with increasing concentrations of dietary CP, was not mediated through decreased enzyme activity associated with the catabolism of methionine, but was the result of an increase in food (methionine) intake.     

Comparison of amino acid profiles between rats subjected to forced running and voluntary running exercises

It has been suspected that in comparison with glucose or fatty acids, the levels of amino acids may readily change with different forms of exercise. In the present study, we measured the concentrations of amino acids, glucose, triglycerides, total protein and total cholesterol in the blood and/or cerebrospinal fluid (CSF) of rats subjected to forced running exercise on a treadmill, and voluntary running exercise using a wheel, with a constant running distance of 440 m. Rats that performed no running and rats subjected to immobilization stress were used as controls. We observed a few significant changes in the levels of plasma glucose, triglycerides, total protein and total cholesterol in all groups. Whereas, plasma amino acid levels were significantly changed by exercise and stress, especially during the light period. The plasma levels of many amino acids were specifically increased by forced running; some were decreased by immobilization stress. Few amino acids showed similar changes in their levels as a result of voluntary running. In addition, there was a significant difference in the degree of amino acid imbalance between blood and CSF. These results provide the first information on changes in levels of amino acids in plasma and CSF resulting from forced and voluntary exercises.     

Characterization of the cDNA and genomic DNA sequence encoding for the platelet integrin alpha IIB and beta III in a horse with Glanzmann thrombasthenia

Glanzmann thrombasthenia (GT) is characterized by a defect of platelet aggregation. This autosomal recessive genetic disorder is caused by an abnormality of the platelet glycoprotein receptors alpha IIb or beta III. Recently, we identified a horse with clinical and pathological features of GT. The aim of this study was to describe this case of GT at the molecular level. A point mutation from G to C in exon 2 of ITGA2B causing a substitution of the expected amino acid arginine 72 (Arg(72)) by a proline (Pro(72)) was encountered. This amino acid change may result in abnormal structural conformations that yield an inactive alpha IIb subunit. The genomic DNA analysis showed that this horse was homozygous for the missense mutation.     

‘二造龙眼’花中氨基酸含量分析

本文针对龙眼特异资源‘二造龙眼’,首次测定分析了其新鲜龙眼花中氨基酸组成和含量。结果显示,‘二造龙眼’鲜花中17种氨基酸种类比较齐全,总氨基酸含量为3234.97 mg?(100g?FW) -1;含量最高的为谷氨酸,最低的为蛋氨酸,两者相差121倍。其中必需氨基酸含量占总氨基酸含量的39.6%,鲜味氨基酸占21.7%,药用氨基酸占61.2%。研究表明,龙眼花氨基酸组成比较合理,具有作为保健食品原料的开发潜力。     

Molecular cloning and expression of the canine metallothionein-III gene.

We have isolated and determined the complete nucleotide sequence of canine metallothionein-III (MT-III) cDNA. The predicted amino acid sequence of the canine MT-III showed a high homology (93%, 87% identity) to that of human and mouse MT-III. The canine MT-III had 2 insertions relative to known mammalian MT-I and MT-II: a threonine after the 4th amino acid and a block of 6 amino acids near the carboxyl terminus. Expression of the canine MT-III mRNA was found exclusively in the central nervous system, where neurons in the olfactory bulb, hippocampus and cerebral cortex showed predominant signals.     

Nicarbazin stimulates intestinal transport of L-leucine in rabbit

Nicarbazin is an anticoccidial drug used mainly in birds, but also in rabbits. Besides the anticoccidial activity, riicarbazin has shown several other effects such as inhibition of growth and feed efficiency in poultries, and stimulation o f sugar intestinal absorption in rabbits. The present work has been performed in order to check whether nicarbazin also affects L-leucine intestinal absorption. The results obtained show that nicarbazin decreases L-leucine accumulation in the jejunal tissue, and increases mucosal to serosal transepithelial fluxes o f this amino acid in a dose-dependent way, without modifying its diffusion across the intestinal epithelium. The drug stimulates the amino acid uptake in brush-border and basolateral membrane vesicles, thus suggesting that nicarlxizin increases the absorption of L-leucine mediated by carriers.     

Immunohistochemical studies on canine cerebral amyloid angiopathy and senile plaques.

Amyloid protein was isolated from the cerebral meninges of 4 aged dogs with cerebral amyloid angiopathy. By immunoblot analysis, antiserum against synthetic oligo-peptide consisting of 1-28 amino acid of amyloid beta protein recognized prominent wide band ranging from 14 to 18 kilodalton (kd). When amyloid samples were solubilized by formic acid, the antiserum recognized lower molecular weight band ranging from 3 to 4 kd. Immunohistochemical studies on cerebral amyloid angiopathy and senile plaques were performed in 17 aged dogs. Anti-amyloid beta protein serum labeled amyloid deposits in cerebral vessel walls and senile plaques. Compact deposits of beta protein were detected in primitive or classical plaques. After using formic acid pretreatment, diffuse deposits of beta protein in the neuropil representing diffuse plaques were detectable. Classical and primitive plaques reacted with antiserum against glial fibrillary acidic protein, while not with antisera against alpha 1-antichymotrypsin, IgG and IgM. Amyloid deposits in the intestines of aged dogs examined, did not react with anti-amyloid beta protein serum.     

鸡传染性法氏囊病病毒弱毒株在鸡体连续传代后毒力增强的分子机理研究

将两株鸡传染性法氏囊病病毒（IBDV）弱毒株在SPF鸡体内连续传代至第5、第6代时,出现明显的法氏囊萎缩和B：B指数下降,表明IBDV弱毒株在鸡体内连续传代后毒力增强。为进一步阐释哪些基因位点导致了上述毒力的变化,本试验测定了基础弱毒株及其在鸡体内传代后各个代次毒的基因组序列,比对分析后发现VP2蛋白253位氨基酸发生了由H到Q或N的变异,表明VP2蛋白253位氨基酸的替换可能会增强传染性法氏囊病病毒在鸡体内的致病性。     

Dietary amino acids fed in free form and as protein components do not differently affect postprandial plasma insulin, glucagon, growth hormone and corticosterone responses in rats

This study examined, whether the postprandial fate of dietary amino acids from different amino acid sources is regulated by the responses of insulin, glucagon, corticosterone and growth hormone (GH). Male Wistar rats were cannulated in the vena jugularis and assigned to dietary groups. The diets contained 21% casein or the same amino acids in free form. In the free amino acid diets, methionine level was varied between the groups. The feed was supplied in two distinct meals. In previous experiments it was established that oxidative amino acid losses of the free amino acid diets and protein diets were different. After 3 weeks on those diets, it appeared that the differences in postprandial oxidative losses had been diminished. GH was measured every 12 min, from 144 min before the start of the experimental meal over the following 144 min. Insulin and corticosterone were measured six times from the start of the meal until 270 min after the meal. No differences have been observed between the hormonal responses to both meals at day 5 and at day 26. In conclusion, it has been found that the differences in the oxidative losses between protein and free amino acid meals are not mediated by the combined action of the insulin, glucagon, corticosterone and GH. Postprandial catabolism of amino acids is most probably regulated by substrate induction.     

Maple syrup urine disease in five hereford calves in ontario

Five newborn grade horned or polled Hereford calves in Ontario exhibited the clinical signs characteristic of hereditary neuraxial edema, namely, inability to rise, and hyperesthesia with extensor spasms upon tactile or auditory stimulation. Histologically, there was widespread spongy vacuolation in both white and gray matter in the brains of all five calves, especially in the midbrain, brain stem, and cerebellum. Amino acid analysis of formalin-fixed cerebral tissue of three calves demonstrated an amino acid composition consistent with a diagnosis of branched chain keto acid decarboxylase deficiency. This condition has been recently reported in Hereford calves in Australia; it is characterized by the delayed onset of neurological signs and severe lesions of status spongiosus, and has been considered analogous to maple syrup urine disease of children.     

标准回肠消化率在肉鸡饲料原料氨基酸营养价值评定中的应用

为了获得最佳生产性能,配制满足家禽营养需要的日粮配方和将排泄到环境中的过多养分降到最低,已显得尤为重要。关于蛋白质饲料原料,多通过表观回肠消化率反映其利用率。以表观回肠消化率值为基础配制日粮,低估了饲料原料的氨基酸消化率,导致排泄物中养分含量增加。通过内源氨基酸基础损失校正表观回肠消化率所得的标准回肠消化率,能克服表观回肠消化率的局限性。作者讨论了标准回肠氨基酸消化率和内源氨基酸基础损失的估测方法,并对其在肉鸡饲料原料氨基酸营养价值评定中的应用作一总结。