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.     

Serum 5'nucleotidase activity in rats: a method for automated analysis and criteria for interpretation

A manual kit for determining serum 5'nucleotidase (5'NT, EC 3.1.3.5) activity was adapted for use with rat samples on a large discrete clinical chemistry analyzer. The precision of the method was good (within-run C.V. = 2.14%; between-run C.V. = 5.5%). A comparison of the new automated method with a manual and semi-automated method gave regression statistics of y = 1.18X -3.66 (Sy. x = 4.54), and y = 0.733X + 1.97 (Sy. x = 1.69), respectively. Temperature conversion factors provided by the kit manufacturer for human samples were determined to be inaccurate for converting results from rat samples. Analysis of components contributing to normal variation in rat serum 5'NT activity showed age and sex to be major factors. Increased serum 5'NT activity was observed in female rats when compared to male rats beginning at about 5 to 6 weeks of age. An analysis of variance of serum 5'NT, alkaline phosphatase, and GGT activities observed over a 9-week period in normal rats suggests several advantages for 5'NT as a predictor of biliary lesions in rats.     

Impact of environmental factors on infestation of tea leaves byHelopeltis theivora, and associated changes in flavonoid flavor components and enzyme activities

Attack of tea plant (Camellia sinensis (L.) O. Kuntze) leaves by the tea mosquito bugHelopeltis theivora (Hemiptera: Miridae) was positively correlated to temperature and rainfall, and partially to humidity, as determined in 12 varieties during the period 2000–2002. The insect attack was severe during the months of May–September, which had high temperature and rainfall, and led to severe loss of biomass due to curling and drying up of the leaves. The biochemical response of these 12 varieties of tea to attack by the insect was determined with special reference to oxidative enzymes and flavonoid flavor components. Insect attack led to an increase in the activities of the oxidative enzymes peroxidase, ascorbate peroxidase and polyphenol oxidase. Activities of phenyl alanine ammonia lyase generally decreased as a result of insect attack, which was significant in the United Planters Association of South India (UPASI) varieties. A significant decrease in polyphenols was also obtained in UPASI varieties. HPLC analysis of catechins revealed a decrease in some of the catechins in the infected leaves. Analysis of theaflavins from infusion of healthy andHelopeltis-infested tea leaves revealed no changes. http://www.phytoparasitica.org posting Dec. 19, 2004.     

In ovo embryotoxicity of α-endosulfan adversely influences liver and brain metabolism and the immune system in chickens

There is a lack of laboratory-based embryonic chicken toxicity studies with the ecologically relevant low dose/s of endosulfan that utilizes a more practical approach such as the chorioallontoic membrane (CAM) injection. In this investigation, 2μg AR grade α-endosulfan/egg (40% of LD₅₀ for embryos) was injected through the CAM in 12-day-old chicken embryos and the activities of glucose-6-phosphatase (G6Pase, EC 3.1.3.9), fructose 1,6-diphosphatase (FDPase, EC 3.1.3.11), adenosine triphosphatase (ATPase, EC 3.6.1.3) and succinic dehydrogenase (SDH, EC 1.3.99.1) and DNA and RNA content in liver and brain tissues and acetyl cholinesterase (AChE, EC 3.1.1.7) in the latter were determined at 24, 48, and 72 h post-exposure. The wet weight of the embryos did not differ between groups. Following endosulfan exposure, except increase in the hepatic ATPase activity (P < 0.01), there was a significant decrease in the following parameters: G6Pase activity in both the liver and brain (P < 0.01), SDH activity in the brain (P < 0.01), brain overall DNA and RNA concentration (P < 0.05), brain AChE activity (P < 0.01). Exposure of 18-day-old embryos to 2-μg endosulfan for 24 h caused decrease (P < 0.01) in the lymphocyte count and IgG content. Histopathology of thymus and bursa of Fabricius revealed a reduction in the population of thymic follicles, smaller thymocytes with the clear vacuoles in cytoplasm and fewer bursocytes accompanied by infiltration of erythrocytes in lymphoid follicles of the endosulfan-treated embryos. It was inferred that in ovo injection of 0.041 μg/g egg weight of α-endosulfan suppress gluconeogenesis (main energy source in embryonic life), nerve transmission, and immunity.     

番茄不同部位中糖含量和相关酶活性的研究

 试验将番茄光合产物运转途径上叶片(源) 、运输系统以及果实(库) 区分开, 分别测定其糖的组成和含量以及糖代谢相关酶的活性。结果表明: 番茄光合产物运转途径上从“源”到“库”各部位糖的组成和含量不同。叶肉中果糖的含量最高, 蔗糖的含量最低; 中筋中以果糖和葡萄糖为主; 叶柄维管束中葡萄糖含量最高, 蔗糖含量次之, 果糖含量最低。节间和果柄维管束中主要含有蔗糖。果实维管束以及果实内各部位中则主要含有葡萄糖和果糖, 且两者含量无显著差异, 蔗糖含量很低。萼片中葡萄糖含量最高, 蔗糖含量最低; 果蒂中3种糖含量均较高且无显著差异。番茄叶肉及光合产物运转组织中转化酶活性很低, 而在库器官的非维管组织中转化酶活性较高。果蒂中的蔗糖合成酶( SS) 活性最高, 其次是叶肉和运转组织, 果实内各部位中SS活性较低。在合成蔗糖的器官—叶肉中, 有较高的蔗糖磷酸合成酶( SPS) 活性, 运转组织中的SPS活性较叶肉中降低, 但果柄维管束和果实维管束中则表现出较高SPS活性, 果肉、果胶质胎座及心室隔壁中的SPS活性最低。     

Ultrastructural Localization of Hydrogen Peroxide in Host Leaves Treated with AK-toxin I Produced by Alternaria alternata Japanese Pear Pathotype

The rapid generation of reactive oxygen species (ROS), called the oxidative burst, is one of the earliest host responses to pathogen infection or elicitor treatments. Therefore, we looked for the induction of ROS generation in Japanese pear leaves by the host-specific toxin, AK-toxin I using a cytochemical method for detecting H₂O₂. A small amount of non-specific generation of H₂O₂ was found in the cell walls in toxin- and water-treated susceptible and resistant leaves. Thus, the generation of H₂O₂ at cell walls appears to be caused by wounding stress during sampling. Specific generation of ROS, however, was found only in the membrane fragments and extended desmotubules characteristic of modified sites of the plasma membrane in the toxin-treated susceptible leaves. In addition, generation of H₂O₂ at plasma membranes was observed with higher frequency in toxin-treated susceptible leaves. This result indicates that the H₂O₂ generation was associated with damaged sites in the plasmalemma after toxin treatment and perhaps with the formation of membrane fragments from altered portions of the invaginated plasma membrane. Received 21 September 2001/ Accepted in revised form 25 October 2001     

Potato diseases caused by soft rot erwinias: an overview of pathogenesis

Three soft rot erwinias, Erwinia carotovora ssp. carotovora , E. carotovora ssp. atroseptica and E. chrysanthemi are associated with potatoes causing tuber soft rot and blackleg (stem rot). Latent infection of tubers and stems is widespread. As opportunistic pathogens, the bacteria tend to cause disease when potato resistance is impaired. Pathogenesis or disease development in potato tubers and stems is discussed in terms of the interaction between pathogen, host and environment, microbial competition and recent findings on the molecular basis of pathogenicity. Emphasis is placed on the role of free water and anaerobiosis in weakening tuber resistance and in providing nutrient for erwinias to multiply. Blackleg symptoms are expressed when erwinias predominate in rotting mother tubers, invade the stems and multiply in xylem vessels under favourable weather conditions. Soft rot erwinias tend to out-compete other bacteria in tuber rots because of their ability to produce larger quantities of a wider range of cell wall-degrading enzymes. However, despite extensive studies on their induction, regulation and secretion, little is known about the precise role of the different enzymes in pathogenesis. The putative role of quorum-sensing regulation of these enzymes in disease development is evaluated. The role certain pathogenicity-related characters, including motility, adhesion, siderophores, detoxifying systems and the hrp gene complex, common to most bacteria including symbionts and saprophytes, could play in latent and active infections is also discussed.     

Functional Analysis of ABC Transporter Genes From Botrytis cinerea Identifies BcatrB as a Transporter of Eugenol

The role of multiple ATP-binding cassette (ABC) and major facilitator superfamily (MFS) transporter genes from the plant pathogenic fungus Botrytis cinerea in protection against natural fungitoxic compounds was studied by expression analysis and phenotyping of gene-replacement mutants. The expression of 11 ABC (BcatrA–BcatrK) and three MFS genes (Bcmfs1, Bcmfs2 and Bcmfs4) was studied. All genes showed a low basal level of expression, but were differentially induced by treatment with cycloheximide and the plant defence compounds camptothecin, eugenol, psoralen, resveratrol and rishitin. The latter compounds induced expression of BcatrB at a high level. Eugenol was more toxic to BcatrB gene-replacement mutants than to the control isolates. Eugenol also caused an instantaneous increase in mycelial accumulation of the fungicide fludioxonil, a known substrate of BcatrB. However, there was no difference in virulence between the wild-type and BcatrB gene-replacement mutants on Ocimum basilicum, a plant known to contain eugenol. The results indicate that BcatrB is a transporter of lipophilic compounds, such as eugenol, but its role in virulence remains uncertain.     

NaCl胁迫下枸杞愈伤组织活性氧产生与质膜H+—ATPase活性的关系

以枸杞愈伤组织为材料,研究了盐胁迫对活性氧伤害和质膜H -ATPase活性的影响。结果表明,NaCl浓度为100mmol/L时超氧阴离子(O2-·)和过氧化氢(H2O2)含量上升,质膜H -ATPase活性先升后降;质膜相对透性与丙二醛(MDA)含量呈显著正相关;超氧阴离子(O2-·)和过氧化氢(H2O2)含量与质膜H -ATPase活性在重度胁迫下呈显著负相关,说明盐胁迫下活性氧积累可能是加速伤害质膜功能的主要原因之一。     

温湿度调控对番茄灰霉病菌产生的细胞壁降解酶的影响

 番茄灰霉病菌在致病过程中能够产生4种细胞壁降解酶,以PMG酶活性最高,其次是β-葡萄糖苷酶和PG酶,Cx最少。灰霉病菌在不同温度下侵染番茄叶片时产生的致病酶活性不同,4种酶在20℃时表现了最高的活性,15℃次之,当温度达到25℃时,各种酶的活性都急剧下降,随着温度的再升高,酶活更低。随着湿度的增高,病菌产生的细胞壁降解酶的活性也增加,当相对湿度达到90%以上时,4种酶的活性也达到最高。温湿度对番茄灰霉病菌产生细胞壁降解酶的影响趋势,与其对发病的影响趋势是一致的。