Using the indicator amino acid oxidation technique to study threonine requirements in horses receiving a predominantly forage diet

Threonine has been reported to be the second limiting amino acid in typical equine diets, but its actual requirement has not been determined in horses. To evaluate amino acid metabolism and requirements, the indicator amino acid oxidation (IAAO) method has been successfully used in other species. The objective of this research was to estimate threonine requirements in mature horses fed timothy hay and concentrate in 4:1 ratio using the IAAO method. Six Thoroughbred mares (579.9 ± 46.7 kg) received each of 6 levels of threonine intake, 41, 51, 61, 70, 80 and 89 mg/kg BW/day, in a randomly determined order. Each study period was 7‐day long, and on day 6, blood samples were collected before and 90 min after feeding to measure amino acid concentrations using HPLC. On day 7, horses underwent IAAO procedures, which included a 2‐hr primed, constant intravenous infusion of [¹³C]sodium bicarbonate to measure total CO₂ production and a 4‐hr primed, constant oral administration of [1‐¹³C]phenylalanine to estimate phenylalanine oxidation to CO₂. Blood and breath samples were collected to measure blood [¹³C]phenylalanine, using GC‐MS analysis and breath ¹³CO₂ enrichment, using an infrared isotope analyser. Increasing threonine intake levels did not affect plasma phenylalanine oxidation by the ANOVA test (p > 0.05) but resulted in a linear decrease in phenylalanine oxidation (p = 0.04) without a breakpoint by the orthogonal linear contrast. This study is the first attempt to evaluate threonine requirements in horses by the IAAO method; however, threonine requirements are still unknown in mature horses at this time.     

苏氨酸和异亮氨酸对皖西白鹅屠宰性能、器官指数及肠道形态结构、消化酶活性、抗氧化和免疫指标的影响

本试验旨在研究苏氨酸和异亮氨酸对皖西白鹅屠宰性能、器官指数及肠道形态结构、消化酶活性、抗氧化和免疫指标的影响。选取30日龄体重相近的皖西白鹅180只,随机分为3组,每组3个重复,每个重复20只。对照组饲喂基础饲粮,苏氨酸组在基础饲粮中添加2.0 g/kg的L-苏氨酸,异亮氨酸组在基础饲粮中添加2.5 g/kg的L-异亮氨酸。试验期60 d。结果表明：1）各组之间90日龄屠宰性能和器官指数无显著差异（P>0.05）。2）异亮氨酸组60日龄十二指肠和空肠绒毛高度、隐窝深度显著或极显著高于对照组（P<0.05或P<0.01）,苏氨酸组60日龄十二指肠、空肠和回肠隐窝深度及90日龄十二指肠和回肠绒毛高度显著或极显著高于对照组（P<0.05或P<0.01）。3）苏氨酸组和异亮氨酸组60、90日龄十二指肠胰蛋白酶活性极显著高于对照组（P<0.01）,苏氨酸组60、90日龄十二指肠麦芽糖酶活性及90日龄空肠淀粉酶、乳糖酶活性显著或极显著高于对照组（P<0.05或P<0.01）。4）异亮氨酸组60日龄空肠总抗氧化能力显著高于对照组和苏氨酸组（P<0...     

猪链球菌丝氨酸/苏氨酸激酶基因缺失株的构建及其致病性

为了研究丝氨酸/苏氨酸磷激酶(STK)在猪链球菌致病过程中的作用,利用大肠杆菌-猪链球菌穿梭质粒p SET4s构建STK基因缺失株△stk。结果显示,stk基因缺失后,缺失株△stk易形成长链状结构;缺失株的半致死剂量(LD50)较亲本株高7倍,对CD1小鼠的致病能力降低;体内定植试验结果表明缺失株△stk在小鼠体内各器官的定植能力显著降低。这些数据表明STK与猪链球菌2型(SS2)的致病性相关。     

陆地棉一个丝氨酸/苏氨酸激酶蛋白基因的克隆与表达分析

通过RT-PCR获得一个编码棉花丝氨酸/苏氨酸激酶类似蛋白全长的cDNA片段,其编码的氨基酸序列与拟南芥ATPK3的相似度达到82%,这个基因暂被命名为GhPK1。基因全编码区包括1038个核苷酸,编码346个氨基酸的蛋白。GhPK1的编码产物在265~270个氨基酸处有一个跨膜区并可能结合在内质网膜上。GhPK1在种子和纤维中的表达水平较其它组织高。另外,GhPK1的表达量在受到盐胁迫后上升,说明GhPK1可能参与了盐胁迫反应。     

22~42日龄黄羽肉鸡饲粮苏氨酸需要量研究

【目的】通过研究饲粮苏氨酸水平对22~42日龄黄羽肉鸡生长性能、胴体品质和免疫功能的影响,确定其苏氨酸需要量。【方法】试验选用22日龄快大型岭南黄羽肉鸡1 600只,根据体重相同的原则分为5个组,每组8个重复,每个重复40只肉鸡（公母各半）。采用玉米-花生粕型基础饲粮,5个处理组试验鸡分别饲喂含0.60%（基础饲粮）、0.67%、0.74%、0.81%和0.88%苏氨酸饲粮,试验期为21 d。试验结束后,每个重复选取2只鸡屠宰采样,测定屠宰性能、免疫器官指数和血清生化指标。【结果】饲粮苏氨酸水平显著影响黄羽肉公鸡、母鸡的体重、平均日增重和料重比（P<0.05）。对于母鸡,0.88%苏氨酸水平组平均日增重最高;0.81%苏氨酸水平组平均日采食量最高。对于公鸡,0.81%苏氨酸水平组平均日增重最高;0.81%苏氨酸水平组平均日采食量最高。饲粮苏氨酸水平与黄羽肉公鸡平均日增重、平均日采食量以及公、母鸡料重比呈显著的二次线性相关（P<0.05）;根据平均日增重和平均日采食量回归方程得出黄羽肉公鸡饲粮苏氨酸最适水平分别为0.81%和0.83%;根据料重比回归方程得出黄羽肉母鸡和公鸡饲粮苏氨酸最适水平分别为0.83%和0.86%。饲粮苏氨酸水平对黄羽肉公鸡、母鸡的全净膛率、半净膛率、腿肌率、胸肌率均无显著影响（P>0.05）;对黄羽肉公鸡全净膛率和腹脂率均有显著影响,其中0.81%苏氨酸组全净膛率显著高于0.60%和0.74%苏氨酸组,0.88%苏氨酸组腹脂率显著低于0.60%和0.67%苏氨酸组（P<0.05）。饲粮苏氨酸水平显著影响黄羽肉母鸡法氏囊指数和胸腺指数,其中0.60%苏氨酸组法氏囊指数显著低于0.81%苏氨酸组,0.81%和0.88%苏氨酸组胸腺指数显著高于0.60%苏氨酸组（P<0.05）;对肝脏指数、脾脏指数无显著影响（P>0.05）。饲粮苏氨酸水平对黄羽肉母鸡血清中甘油三酯、胰岛素含量有显著影响,其中0.60%苏氨酸水平组血清中胰岛素含量最低,0.88%苏氨酸水平组血清中甘油三酯含量最低,显著低于0.60%苏氨酸组（P<0.05）;苏氨酸水平对黄羽肉公鸡甘油三酯含量有显著影响,其中0.88%苏氨酸水平组血清中甘油三酯含量最低,显著低于0.60%和0.67%苏氨酸组（P<0.05）。【结论】饲粮中添加适量苏氨酸可以提高黄羽肉鸡生长性能、胴体品质,促进免疫器官发育。以生长性能为主要判定指标,确定22~42日龄黄羽肉母鸡饲粮最适苏氨酸水平为0.83%,黄羽肉公鸡最适苏氨酸水平为0.81%。     

Effects of STK15 overexpression on growth of human esophageal carcinoma cell line KYSE150

AIM:To investigate the effects of serine/threonine kinase 15 (STK15) overexpression on the growth of human esophageal squamous-cell carcinoma (ESCC) cell line KYSE150. METHODS:Recombinant pEGFP-C1-STK15 expression vector was transfected into KYSE150 cells using LipofectamineTM 2000 and the expression of STK15 was detected by fluorescence microscopy and Western blotting. The proliferation of the cells in vitro was measured by MTT assay. The cell cycle distribution and apoptosis were detected by flow cytometry. The proliferation of the cells in vivo was measured by tumorigenicity experiment in nude mice. RESULTS:After recombinant pEGFP-C1-STK15 expression vector was stably transfected into KYSE150 cells, GFP-STK15 fusion protein localized to centrosome and spindle. The STK15-overexpressing colonies were further confirmed by Western blotting. MTT assay showed that the proliferation of the cells in STK15 overexpression group was increased compared with control group (P<0.01). Flow cytometry analysis showed that the percentage of the cells in G0/G1 phase and the cell apoptosis in STK15 overexpression group were decreased compared with control group (P<0.01). The tumorigenicity experiment in nude mice showed that the proliferation of the cells in STK15 overexpression group was increased compared with control group (P<0.01). CONCLUSION: Overexpression of STK15 in human ESCC KYSE150 cells promotes the cell growth in vitro and in vivo, indicating that STK15 may serve as a novel therapeutic target for esophageal carcinoma.     

苏氨酸对体外培养感染伪狂犬病毒猪空肠上皮细胞免疫相关基因表达的影响

本研究旨在探讨苏氨酸对体外培养感染伪狂犬病毒(PRV)猪空肠上皮细胞免疫相关基因表达的影响.试验选用IPEC-J2细胞为细胞模型,分为4个处理,对照组为未经PRV感染、苏氨酸浓度53.45 mg/L,Ⅰ、Ⅱ、Ⅲ组为试验组,均经PRV感染,苏氨酸浓度分别为53.45、106.90、160.35 mg/L,每组3个重复,每个重复4孔.37℃、5％ CO2培养后1、6、12、24 h每组取1个重复采集细胞,用实时荧光定量PCR方法检测白介素1β(IL-1β)、白介素6(IL-6)、白介素15(IL-15)、肿瘤坏死因子α(TNF-α)及转化生长因子β1( TGF-β1) mRNA表达量.结果表明:1)PRV感染极显著降低了IL-1β mRNA表达量(P =0.001),苏氨酸水平(P=0.830)、时间与苏氨酸水平交互作用(P =0.249)的影响均不显著;2)PRV感染对IL-6mRNA表达量影响不显著(P =0.162),随苏氨酸水平升高,IL-6 mRNA表达量极显著地先升高后降低(P=0.002),时间与苏氨酸水平交互作用的影响显著(P =0.012);3)PRV感染极显著提高了IL-15 mRNA表达量(P =0.000),提高苏氨酸水平极显著提高了IL-15 mRNA表达量(P=0.000),时间与苏氨酸水平交互作用的影响亦极显著(P =0.000);4) PRV感染极显著提高了TNF-α mRNA表达量(P=0.000),苏氨酸水平(P =0.113)、时间与苏氨酸水平交互作用(P =0.195)的影响均不显著;5)PRV感染极显著提高了TGF-β1 mRNA表达量(P=0.000),时间与苏氨酸水平交互作用的影响显著(P =0.015),苏氨酸水平的影响不显著(P=0.055).结果提示,补充苏氨酸对感染PRV的IPEC-J2细胞先天性免疫功能具有分子表达水平的调控作用,总体上能够抑制IL-1β、TNF-α、TGF-β1基因表达,加强IL-6、IL-15基因表达,但影响具有时间效应.     

A deficient or an excess of dietary threonine level affects intestinal mucosal integrity and barrier function in broiler chickens

The aim of this study was to investigate the effects of deficient or excess of dietary threonine (Thr) levels on intestinal integrity and barrier function of broilers. A total of 432 1‐day‐old commercial broilers (Arbor Acre) were assigned to four experiment groups consisting of six replicates of 18 birds. The treatments were designed as follows: 85%, 100%, 125% and 150% of NRC (Nutrient requirements of poultry (9th edn). Washington, DC: The National Academies Press, 1994) recommendations. The results indicated that expressions of jejunal and ileal secretory immunoglobulin A (sIgA) mRNA were increased linearly or quadratically by increasing Thr (p < .05), and the highest sIgA mRNA abundance was obtained in 125% Thr level. Likewise, the intestinal sIgA content showed similar increasing trend with the intestinal sIgA gene expression in this instance. The high level of Thr inclusion upregulated mucin 2 (MUC2) mRNA expression in the jejunum and ileum (p < .05). In addition, on day 21, the expression levels of jejunal zonula occludens‐2 (ZO‐2) and ileal zonula occludens‐1 (ZO‐1) decreased then increased with increasing Thr level (p < .05), whereas, the mRNA expressions of occludin in the jejunum and ileum had no significant difference amongst groups (p >.05). On day 42, Thr treatments did not affect the mRNA abundance of measured genes in the jejunum and ileum (p > .05). These findings suggested that Thr might be a nutrient immunomodulator that affects intestinal barrier function, moreover, 125% of the NRC (1994) recommendations Thr level was optimum.     

Dietary threonine requirement of fingerling Indian major carp,Catla catla (Hamilton) estimated by growth,protein retention efficiency,threonine deposition,haematological parameters and carcass composition

A 12‐week feeding trial was conducted to determine the dietary threonine requirement of fingerling Indian major carp, Catla catla (3.35 ± 0.11 cm; 0.59 ± 0.06 g). Six casein‐gelatin based (33% crude protein; 3.23 kcal g^?1 digestible energy) amino acid test diets with graded levels of analysed threonine (0.74%, 0.96%, 1.21%, 1.48%, 1.72% and 1.93% dry diet) were fed to satiation to triplicate groups of fish. Absolute weight gain (g per fish), feed conversion ratio, protein retention efficiency, threonine deposition, RNA/DNA ratio and carcass protein significantly improved with the increase in dietary threonine and peaked at 1.48% of the dry diet. Haematological indices were also found to be best in fish fed at 1.48% threonine diet. Quadratic regression analysis of absolute weight gain, feed conversion ratio, protein retention efficiency, threonine deposition, RNA/DNA ratio, carcass protein, haemoglobin (g dL^?1), haematocrit (%) and RBCs (10⁶ × mm^?3) at 95% of maximum and minimum response exhibited the threonine requirement of fingerling C. catla between 1.35% and 1.48% dry diet, corresponding to 4.09–4.48% dietary protein. Present finding would be useful in formulating threonine‐balanced feeds for the intensive culture of C. catla.