Advanced oxidation protein products are generated by bovine neutrophils and inhibit free radical production in vitro

Despite the recognised importance of oxidative stress in the health and immune function of dairy cows, protein oxidation markers have been poorly studied in this species. The current study aimed to characterise markers of protein oxidation generated by activated bovine neutrophils and investigate the biological effects of advanced oxidation protein products (AOPP) on bovine neutrophils. Markers of protein oxidation (AOPP, dityrosines and carbonyls) were measured in culture medium containing bovine serum albumin (BSA) exposed to neutrophils. The effect of AOPP-BSA on generation of reactive oxygen species (ROS) was assessed by chemiluminescence. Activation of caspases-3, -8 and -9 and the presence of DNA laddering were used as apoptosis markers.Greater amounts of AOPP were generated by phorbol myristate acetate (PMA)-activated than non-activated neutrophils (1.46 ± 0.13 vs. 0.75 ± 0.13 nmol/mg protein, respectively; P < 0.05). Activated neutrophils and hypochlorous acid generated slightly different patterns of oxidized protein markers. Exposure to AOPP-BSA did not stimulate ROS production. Activated neutrophils generated a lesser amount of ROS when incubated with AOPP-BSA (P < 0.001). Activation with PMA induced a loss of viable neutrophils after 3 h, which was greater with AOPP-BSA incubation (P < 0.05). Detectable amounts of active caspases-3, -8 and -9 were found in nearly all samples but differences in caspase activation or DNA laddering were not observed comparing treatment groups. Apoptosis was unlikely to be responsible for the greater loss of PMA-activated neutrophils cultured in AOPP-BSA and it is possible that primary necrosis occurred. The results suggest that accumulation of oxidized proteins at an inflammatory site might result in a progressive reduction of neutrophil viability.     

Effects of different dietary energy and protein levels and sex on growth performance,carcass characteristics and meat quality of F1 Angus x Chinese Xiangxi yellow cattle

Background：The experiment evaluated the effect of nutrition levels and sex on the growth performance,carcass characteristics and meat quality of F1 Angus × Chinese Xiangxi yellow cattle.Methods：During the background period of 184 d,23 steers and 24 heifers were fed the same ration,then put into a2×2×2 factorial arrangement under two levels of- dietary energy（TON：70/80%DM）,protein（CP：11.9/14.3%DM）and sex（S：male/female） during the finishing phase of 146 d.The treatments were-（1） high energy/low protein（HELP）,（2） high energy/high protein（HEHP）,（3） low energy/low protein（LELP） and（4） low energy/high protein（LEHP）.Each treatment used 6 steers and 6 heifers,except for HELP- 5 steers and 6 heifers.Results：Growth rate and final carcass weight were unaffected by dietary energy and protein levels or by sex.Compared with the LE diet group,the HE group had significantly lower dry matter intake（DMI,6.76 vs.7.48 kg DM/d）,greater chest girth increments（46.1 vs.36.8 cm）,higher carcass fat（19.9 vs.16.3%） and intramuscular fat content（29.9 vs.22.8%DM）.The HE group also had improved yields of top and medium top grade commercial meat cuts（39.9 vs.36.5%）.The dressing percentage was higher for the HP group than the LP group（53.4 vs.54.9%）.Steers had a greater length increment（9.0 vs.8.3 cm）,but lower carcass fat content（16.8 vs.19.4%） than heifers.The meat quality traits（shear force value,drip loss,cooking loss and water holding capacity） were not affected by treatments or sex,averaging 3.14 kg,2.5,31.5 and 52.9%,respectively.The nutritive profiles（both fatty and amino acid composition） were not influenced by the energy or protein levels or by sex.Conclusions：The dietary energy and protein levels and sex significantly influenced the carcass characteristics and chemical composition of meat but not thegrowth performance,meat quality traits and nutritive profiles.     

Effects of the protein/carbohydrate ratio of extruded diets on protein synthesis, protein growth and body composition in juvenile brown trout (Salmo trutta)

High levels of protein in fish feeds result in higher costs and nitrogen waste. Therefore, studies focused on protein substitution by alternative energy sources are necessary. Here we examine whether the protein:carbohydrate ratio in extruded diets affects protein-turnover parameters and the main tissue components in brown trout. Juvenile fish were adapted to two extruded diets: one higher in protein and lower in carbohydrate, C: 45/28 and the other higher in carbohydrate and lower in protein, HC: 37/40. Gross energy was 19 MJ kg⁻¹ DM in both diets. Protein and lipid digestibility were high and similar in both groups (≈90%), whereas total and carbohydrate digestibility and digestible energy were lower in the HC-fed group than in the C-fed group. Consequently, when fish were adapted to the diets, plasma levels of glucose and insulin differed between diets. HC-group presented higher plasma glucose levels and lower plasma insulin levels than C-group. Protein synthesis rates in white muscle, liver and whole body did not differ significantly between diets. In contrast, protein accretion rate of white muscle and whole body were significantly lower in the HC group, indicating an increase of protein degradation in these tissues and a decrease of synthesis retention efficiency. In spite of the daily protein intake of HC group was lower than C-group, the anabolic stimulation efficiency was increased by 34% in HC group. Protein and lipid contents in white muscle and liver were stable throughout the experiment. No hepatomegalia or increased fat deposition was observed in fish fed HC. Differences in specific growth rates (C: 0.88%; HC: 0.77%) were associated more with the lower protein consumption rate and the lower level of digestible energy in fish fed HC than with the higher dietary carbohydrate content of the diet.     

The effect of dietary protein source on growth and carcass composition in juvenile Australian freshwater crayfish

A feed trial was conducted for 12 weeks on juvenile Australian freshwater crayfish (Cherax destructor) (mean weight (SE) 0.82 (0.02)g) maintained on five isoenergetic diets with a protein content of 30%. Diets differed in the primary source of protein used, with meat, snail, soybean, yabby, and zooplankton meals comprising the major protein ingredient, varying from 56–60% of total protein. Mean percentage weight gain per day ranged from 7.57% (yabby meal diet), to 9.42% (snail meal diet). No significant difference occurred in mean weight, percentage weight gain, specific growth rate (%), or survival among diets. A maximum size of 16.44g was attained on the yabby meal diet. Largest mean weight was 8.27g on the snail-based diet. Food conversion ratios were all good, with a minimum value of 0.95 recorded for the snail-based diet. Initial food consumption per day was approximately 5% of mean animal weight, decreasing to around 2.4%, and is collectively described by a power curve. Protein retention ranged from 29.57% in the zooplankton meal diet to 41.15% in the snail-based diet. Carcass composition was influenced by feed type, with the most marked difference occurring in carapace colour. Animals fed the zooplankton-based pellets developed the strongest pigmentation. Recommendations are made for including certain protein-based ingredients in manufactured yabby diets.     

副溶血弧菌的外膜蛋白及其抗原性研究↑（*）

本文对１３株不同来源的副溶血弧菌的外膜蛋白及其抗原性进行了比较研究。１３株副溶血弧菌的外膜蛋白有相似的ＳＤＳ－ＰＡＧＥ图谱。大多数菌株有五条主要的蛋白质，其大致分子量分别为：ａ、６９ｋＤａ；ｂ、６３ｋＤａ；ｃ、４０ｋＤａ；ｄ、３０ｋＤａ；ｅ、２８ｋＤａ。其中蛋白质ｂ是所有菌株共有的。与吸附后的兔抗副溶血弧菌血清Ｗｅｓｔｅｒｎ印迹法结果显示抗血清与多数副溶血弧菌菌株的外膜蛋白有一条共同的免疫反应带，其大致分子量为４４ｋＤａ。     

野生大豆回交导入系蛋白质含量性状的QTL分析

以野生型大豆ZYD00006（供体亲本）与黑龙江省主栽品种绥农14（轮回亲本）所构建的回交导入系（1204株）为研究材料,利用WinQTL2.5的复合区间作图法(CIM)在9个连锁群定位了16个与蛋白质含量相关的QTL（14个正效应,2个负效应）;导入系群体经过严格的蛋白质含量筛选鉴定,得到10个蛋白质含量性状明显大于轮回亲本的导入系株行。利用这10个高蛋白含量株行（选择群体）结合随机对照群体,通过基于遗传搭车原理的卡方分析,检测到分布于10个连锁群上的17个与大豆蛋白质含量相关的标记位点,对蛋白质含量表现为正效应。两种方法共同检测到7个QTL。这些材料和位点将为高蛋白含量相关基因克隆及分子辅助育种提供重要的材料基础和标记信息。     

TaJAZ7D蛋白在冬小麦JA抗寒途径中的作用

转录抑制因子JAZ（Jasmonate ZIM-domain）蛋白是茉莉酸（Jasmonate,JA）信号转导途径的核心元件,前人研究发现,拟南芥中AtJAZ1、AtJAZ4与AtICE1蛋白互作可抑制 AtICE1基因的转录,负调控拟南芥的抗寒性,然而小麦中TaJAZ与TaICE蛋白的互作调控机制尚不清楚。本研究以强抗寒冬小麦品种东农冬麦1号（Dn1）为试验材料,以与拟南芥AtJAZ1、AtJAZ4蛋白高度同源的TaJAZ7、TaJAZ12蛋白为对象,检测外源茉莉酸甲酯（MeJA）对低温胁迫下 TaJAZ7、 TaJAZ12基因表达量的影响。结果发现, TaJAZ7基因的表达量与温度变化呈明显负相关,故对TaJAZ7(以TaJAZ7D为研究对象进行后续研究)蛋白进行生物信息学分析和亚细胞定位,并利用酵母双杂交技术验证TaJAZ7D蛋白与TaMYC2和TaICE41蛋白的互作。生物信息学分析表明, TaJAZ7D基因编码区序列全长为633 bp,为不稳定的亲水性混合型蛋白;TaJAZ7D蛋白有典型的TIFY和Jas结构域,属于TIFY家族。亚细胞定位发现,TaJAZ7D蛋白位于细胞核中。酵母双杂交验证试验发现,TaJAZ7D蛋白与TaMYC2和TaICE41蛋白分别存在互作关系。     

Bipolaris australiensis HD-1胆红素氧化酶纯化、鉴定及其酶学性质

[目的]为了获得B.bipolaris HD-1胆红素氧化酶纯品,确定酶的同源性及其常规酶学性质.[方法]将B.bipolaris HD-1发酵液依次通过硫酸铵盐析、阴离子交换层析和凝胶过滤层析确定蛋白的纯度、浓度和酶活力;通过基质辅助激光解析电离飞行时间质谱（MALDI-TOF）法获得氨基酸序列,并利用Mascot软件在NCBI蛋白库中的比对,以确定同源蛋白来源;用SDS-PAGE方法测其表观分子量;用等电聚焦方法测定等电点;用Linwerver-Burk双倒数作图法测定米氏常数;并对该酶的酶学性质进行了常规的测定.[结果]获得具有胆红素氧化酶酶活性的电泳纯单体蛋白,酶活为46 000 U/L,比活为1.15 U/mg;MALDI-TOF共获得135个氨基酸,该蛋白与来源于Hel-minthosporium tritici-vulgaris的漆酶前体蛋白序列同源性达100％;蛋白的表观分子量为68 kDa;pI为4.1;它对ABTS和胆红素的米氏常数分别为Km（ABTS） =1.8×10-5 mol/L（pH 3.0）和Km（bilirnbin）=2.7×10-5 mol/L（pH 7.5）;对胆红素的最适催化活性为36℃,酶在-40℃下可保持2年,酶活保持在90％以上,在pH 8 ～9.5下可保持相当高的催化胆红素活性,能够耐受高浓度的硝酸钠和尿素,低浓度的叠氮化钠对酶活有促进作用而高浓度则抑制,对氯化钠、溶解氧敏感.[结论]该蛋白具有典型的胆红素氧化酶特性,又存在明显的不同.     

Effect of ischemia or hypoxia on vascular endothelial growth factor production in rat myocardium and its significance

AIM:To determine the relationship between ischemia, hypoxia and the production of vascular endothelial growth factor in rat myocardium and its basic mechanism. METHODS:(1) 28 Wistar rats were randomly divided into 4 groups: group A, normal control;group B, 1 day's acute myocardial infarction;group C, 3 day's acute myocardial infarction;group D, 7 day's acute myocardial infarction. (2) Rat cardiac myocytes cultured were primarily divided into some groups, hypoxia incubated 24 hours; PMA groups, hypoxia incubated 24 hours with PKC activator (PMA), A 0 ng/mL; B 10 ng/mL; C 100 ng/mL; D 1 000 ng/mL; Chelerythrine groups, hypoxia incubated 24 hours with PKC inhibitor (chelerythrine), A 0 nmol/L; B 10 nmol/L. (3) By computer scanned and quantitated, vascular endothelial growth factor (VEGF) protein was detected with immunohistochemical technique. RESULTS:The longer time of ischemia and hypoxia was, the higher the VEGF production.The relat ionship was found between the time of ischemia or hypoxia and the production of VEGF.The product ion of VEGF protein was further promoted by PMA with different concentrat ion, decreased by chelerythrine.CONCLUSION: Ischemia or hypoxia strongly stimulated the production of VEGF in myocardium, which played an important role in autoprotecting of ischemic or hypoxic myocardium. Hypoxia-induced PKC activation is one kind of basic mechanisms in this course.