银杏幼苗叶片转化酶活性及其动力学

采用波钦诺克XH法研究了田间和室内栽培的银杏幼苗叶片转化酶的活性及其动力学性质。结果表明:银杏幼苗叶片转化酶在pH值5.0、温度55℃时活性最强;该酶的米氏常数为3.7×10-2mol·L-1;室内盆栽的银杏幼苗叶片转化酶的活性显著高于田间栽培的同龄银杏幼苗叶片转化酶的活性。     

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。     

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.     

长期培肥对土壤酶活性的影响

25a的有机肥长期定位试验地中土壤养分和有关酶活性研究表明:耕种和不同的培肥方式均能提高土壤养分的含量,明显地改善土壤的生物活性,其中对土壤脲酶和磷酸酶活性影响明显,多酚氧化酶显著性则较差。对土壤酶活性进行相关分析发现,土壤脲酶和碱性磷酸酶之间呈极显著正相关,且这两种酶与有关主要土壤肥力因子均呈显著或极显著正相关关系。揭示出了土壤脲酶和碱性磷酸酶活性可以用来作为评价土壤肥力的综合指标。     

1-(1-苯基-1,2,4-三氮唑-3-氧代乙酰氧基)烃基膦酸酯的合成与植物生长调节活性

合成了一系列新的1-(1-苯基-1, 2, 4-三氮唑-3-氧代乙酰氧基) 烃基膦酸酯, 并测定了它们对小麦和黄瓜的植物生长调节活性。结果表明, 在100 Lg/g 时, 所有化合物对小麦芽鞘的生长具有抑制作用, 而在10 Lg/g时化合物6a、6g 对黄瓜显示了较高的促生长活性。