In vitro expression and inhibition of procoagulant activity produced by bovine alveolar macrophages and peripheral blood cells

Local and systemic activation of coagulation is frequently associated with bacterial sepsis. The coagulopathy is due, at least in part, to expression of tissue factor (TF) by monocytes and macrophages. The purpose of this study was to evaluate the expression of procoagulant activity by bovine alveolar macrophages, leukocytes and platelets, and to determine the relative potency of three chemical inhibitors of TF expression (pentoxifylline, retinoic acid, and cyclosporin A). Bovine alveolar macrophages were stimulated with lipopolysaccharide (LPS) derived from Pasteurella haemolytica or recombinant bovine tumour nervous factor (TNF) and dose- and time-dependent effects on TF expression were studied. LPS and TNF induced TF expression in alveolar macrophages and LPS treatment of whole blood induced TF expression in mononuclear cells. Neutrophils and platelets also expressed procoagulant activity, but this activity was not inhibited by anti-bovine TF monoclonal antibody. Pentoxifylline (40 mol/L), retinoic acid (0.01 mmol/L) and cyclosporin A (0.08 mol/L) inhibited TF expression when added concurrently with LPS or TNF, but not when added 4 h after stimulation. TF mRNA was not detected in unstimulated alveolar macrophages by Northern blot analysis. In contrast, exposure to LPS or TNF for 6 h induced marked expression of TF mRNA, which was inhibited by treatment with pentoxifylline, retinoic acid and cyclosporin A. Expression of TNF by alveolar macrophages stimulated with LPS was also inhibited by these compounds. Our results indicate that procoagulant activity expressed by alveolar macrophages and monocytes is associated with expression of TF, whereas procoagulant activity expressed by neutrophils and platelets is not. The concentrations of pentoxifylline and retinoic acid necessary for inhibition of TF expression in vitro may not be achievable in vivo owing to their toxic effects. However, the in vitro concentration of cyclosporin A that inhibited TF expression did not exceed the plasma concentration observed in humans, and therefore may be useful for inhibition of TF expression in vivo.Abbreviations BAL bronchoalveolar lavage - LPS lipopolysaccharide - cDNA cloned deoxyribonucleic acid - cAMP cyclic adenosine monophosphate - GAPDH glyceraldehyde phosphate dehydrogenase - mRNA messenger ribonucleic acid - TF tissue factor - TNF tumour necrosis factor - DPBS Dulbecco's phosphate-buffered saline     

Serum lipase determination in the dog: a comparison of a titrimetric method with an automated kinetic method

An enzymatic, kinetic method for determining serum lipase activity was evaluated and compared to a standard manual method for use in dogs. The kinetic method was a commercial kit adapted for use on a tandem access clinical chemistry analyzer and utilized a series of coupled enzymatic reactions based on the hydrolysis of 1,2-diglyceride by lipase. The manual method was the Cherry-Crandall technique based on the titration of base against the acid formed by hydrolysis of an olive oil substrate by lipase. The correlation between the two methods was very good (r = 0.94). The reference range for 56 clinically healthy dogs assayed by the kinetic method was 90 to 527 U/L. Diseases associated with a greater than twofold elevation in serum lipase activity as determined by the kinetic method included pancreatitis, gastritis with liver disease, and oliguric renal failure with metabolic acidosis. In some cases, pancreatitis was seen with other clinical problems, such as gastroenteritis, diabetic ketoacidosis, duodenal mass, disseminated intravascular coagulation, and septic peritonitis. Diseases associated with serum lipase activity within the reference range or elevated less than twofold included gastritis, gastric ulcer, cholestasis, phenobarbital-induced hepatopathy, colitis, copper hepatopathy, abdominal hematoma, apocrine gland adenocarcinoma, and thrombocytopenia with pneumonia.     

Trichoderma harzianum produces nonanoic acid, an inhibitor of spore germination and mycelial growth of two cacao pathogens

An isolate of Trichoderma harzianum Rifai from an infected cacao pod produces and secretes nonanoic (pelargonic) acid into a liquid culture medium. Nonanoic acid (NA) was very inhibitory to spore germination and mycelial growth of two cacao pathogens, Crinipellis perniciosa Stahel and Moniliophthora roreri Cif. H.C. Evans. It was highly active causing 75% inhibition of spore germination in an in vitro assay at a rate as low as 0.09 μM for M. roreri and 0.92 μM for C. perniciosa. Mycelial growth was comparatively less sensitive to inhibition, but still there was a 75% reduction in growth with 0.62 μM in M. roreri and 151 μM NA in C. perniciosa. In contrast, NA did not affect Trichoderma mycelial growth or spore germination at concentrations that were inhibitory to the pathogens. 6-pentyl-α-pyrone was also produced and secreted into the medium by T. harzianum, however; it was not antagonistic to the cacao pathogens. Although a number of metabolites produced by Trichoderma spp. have been identified in the past, this is the first report of NA production and secretion by any Trichoderma. The results suggest that NA may play a role in the successful use of some Trichoderma spp. isolates in the biocontrol of fungal diseases of plants.     

Molecular cloning, expression, and characterization of a phi-type glutathione S-transferase from Oryza sativa

The structural gene for glutathione S-transferase in Oryza sativa was successfully cloned from a cDNA library by the polymerase chain reaction method. The deduced amino acid sequence of this gene showed 44-66% similarity to the sequences of the class phi GSTs from Arabidopsis thaliana and Zea mays. This gene was expressed in Escherichia coli with the pET vector system and the gene product was purified to homogeneity by GSH-Sepharose affinity column chromatography. The expressed OsGSTF3-3 was a homo-dimer composed of 24 kDa subunit and its pI value was approximately 7.3. The OsGSTF3-3 was retained on GSH affinity column and its K_m value for GSH was 0.28 mM. The OsGSTF3-3 displayed high activity toward 1-chloro-2,4-dinitrobenzene, a general GST substrate and also had high activities towards acetanilide herbicides, alachlor, and metolachlor. The OsGSTF3-3 was highly sensitive to inhibition by benastatin A and S-hexyl-GSH. From these results, the expressed OsGSTF3-3 is a phi class GST and seems to play an important role in the conjugation of the chloroacetanilide herbicides.     

甜樱桃芽酚类物质含量及相关酶活性变化与自然休眠的关系

 以7年生甜樱桃‘红灯’和‘早红宝石’为试材, 研究了酚类物质含量及相关酶活性在自然休眠期间的动态变化, 探讨了温度对酚类物质含量的影响。结果表明, 甜樱桃芽酚类物质在自然休眠期间持续缓慢增加, 随休眠的结束, 含量急剧下降, 不同品种之间有差异, 自然休眠结束后总酚含量降至最低。花芽中的酚类物质含量略高于叶芽, 多酚氧化酶( PPO) 、苯丙氨酸解氨酶( PAL) 活性差别不大; 不同需冷量的品种之间差异明显。芽中PAL活性与总酚含量的变化趋势呈正相关。而PPO活性在自然休眠期间逐渐升高。低温(5℃) 促进酚类物质的积累, 中期使其提前达到高峰并进入下降阶段, 后期加速其降低;高温(20℃) 效果相反, 变温(5℃/20℃) 效果不明显。     

Effect of the Pietrain breed used as terminal boar on Post-weaning Multisystemic Wasting Syndrome (PMWS) in the offspring in four PMWS-affected farms

The aim of this study was to determine the potential risk factors for PMWS at the individual pig level and assess the effect of the Pietrain paternal genetic background of the animals in a cohort study. The survey was set up in four PMWS-affected farms with 2 repetitions (batches) per farm. A representative sample of 60 pigs per batch, stratified according to the paternal genetic background (Pietrain: yes vs. no), was randomly selected after farrowing. The representative cohort was divided into 8 batches and the pigs were individually monitored from birth to slaughter. Survival analysis was used to determine the factors related to the time to PMWS. The litter-cluster effect was taken into account using the marginal Cox model (robust estimation of the covariance matrix) and the gamma shared frailty model which were compared.No protective effect of the Pietrain breed on the time to PMWS and the proportion of affected pigs in the offspring was found in this study. Piglets showing low circovirus type 2 (PCV2) titres at 7 weeks-old with no subsequent seroconversion and piglets from PCV2 negative sows were most likely to be affected by PMWS (HR=7.0 and 2.8, respectively). Active infection of the pregnant dams with parvovirus was related to an increased risk of PMWS in the offspring (HR=2.3). Neck injuries due to poorly performed injections in the dams were associated with an increased risk of PMWS with the marginal model (HR=2.1). Oxytocin injection (dams) during farrowing was protective against PMWS in the offspring (HR=0.6).     

烫漂及冻结对青头菌营养价值的影响

研究了在不同的烫漂温度和烫漂时间下青头菌过氧化物酶(Peroxidase，POD)活性的变化．烫漂及冻藏对青头菌过氧化物酶活性、氨基酸、矿质元素、维生素及氨基酸态氨等成分的影响结果表明：采用95～96℃烫漂温度效果最好，在4min内过氧化物酶活性降低了95．5％；烫漂及冻藏都使青头菌营养成分受到不同程度的损失建议采用一70℃以下温度不经烫漂而冻结，冻品在-26℃冰箱中贮藏12个月，其营养价值保持很好。     

氮肥对辣椒果实中辣椒素含量的影响

以田间试验和室内分析相结合的方法,采用景尖椒3号(高辣度),以3种不同的氮肥施用量处理,就生长期中氮肥对辣椒果实中辣椒素含量的影响进行研究.结果发现,在一定条件下,增加氮肥施用量会导致辣椒果实中辣椒素含量减少;果实中过氧化物酶活力与辣椒素含量呈一定负相关.     

不同能量水平日粮对肉仔鸡生长及屠宰性能指标的影响研究

选择3周龄健康、无病、体重无显著差异的肉仔鸡96只,随机分为3组,每组设4 个重复,每个重复8只。分别饲喂蛋白质水平(19．5％)相同而能量水平不同(3．0 Mcal／kg、 3．08 Mcal／kg、3．16Mcal／kg)的3种饲粮。试验结果表明:前两周饲料转化(采食／增重)均以 中能量组(3．08 Mcal／kg)最好,为1．4416±0．1199,但三种不同能量水平对肉仔鸡平均日增 重、平均日耗料、饲料转化率均差异不显著(P>0．05);后两周饲料转化仍以中能量组最好,为 1．6362±0．1346,且显著优于高能量组(P<0．05),而与低能组和中能组之间差异不显著,各 能量处理组蛋白质、粗纤维、钙的代谢率均差异不显著(P>0．05),而磷的代谢率却存在显著 差异(P<0．05),其中,以中等能量组的磷代谢率最高,平均为90．73％;3种能量水平下肉仔鸡 半净膛率、全净膛率均差异不显著(P>0．05),表明能量水平并不影响肉仔鸡屠宰性能。     

破壁蜂花粉对蛋鸡免疫功能及肠道菌群的影响

试验将槐花蜂花粉进行振动磨超微粉碎破壁,分别按1 g/kg、5 g/kg、10 g/kg三个不同剂量混到蛋鸡饲料中,从7日龄开始饲喂至50日龄,每个剂量分两个试验,试验一用新城疫Ⅳ系疫苗免疫雏鸡,利用β-微量法监测血清中的血凝抑制抗体效价的动态变化;试验二用大肠杆菌油乳苗免疫雏鸡,利用间接血凝法监测血清中大肠杆菌抗体效价的动态变化。分别定期称重捕杀取脾、胸腺、法氏囊,计算免疫器官指数,取肠道内容物计数大肠杆菌和乳酸菌,测定肠道内pH值。结果表明,不同剂量的破壁蜂花粉均能不同程度地提高病毒抗原性抗体效价和菌体抗原性抗体效价,对雏鸡免疫器官的发育以及肠道内环境、菌群的平衡也有一定影响。