Aktivierung,Aktivität und Inhibierung des Rindertrypsins

Trypsin is a prototype of a large group of enzymes belonging to serine proteinases. The X-ray crystal-structure analyses of its proenzyme trypsinogen, of the active trypsin and of their complexes formed with the pancreatic trypsin inhibitor (PTI) have considerably enhanced our understanding of the mechanisms of activitation, action and inhibition. The trypsinogen is an incompletely folded molecule. Its substrate-binding site becomes only completely fixed upon the enzymatic cleavage of an N-terminal peptide. The contact regions of trypsin and PTI are almost complementary. The complex formed is a (stable) intermediate in the normal tryptic substrate-cleavage reaction.     

长期食入抗坏血酸对国产防锈水诱发威斯塔大鼠致癌作用的影响

经40头雄威司塔大鼠长期饮用国产防锈水(含亚硝酸钠、三乙醇胺和聚乙二醇)实验表明,实验动物每天口服约75mg亚硝酸钠,两年后发生以胰腺泡型腺癌(22.5%)为主的恶性肿瘤(32.5%),并有30%的胰腺泡增生及不典型增生。另40头同规格大鼠在上述处理同时每天还经饮用水口服150mg抗坏血酸,两年后胰腺腺泡增生和胰腺癌发生率均有极显著减少,仅占2.5%,实验提示防锈水诱发胰腺癌是由于在体内合成某种亲胰腺腺泡的未知亚硝基化合物,而口服抗坏血酸对接触“防锈水”的金属切削工人可能有良好的防癌作用。     

Chorionic villus metaphase chromosomes and interphase nuclei analysed by chromosomal in situ suppression (CISS) hybridization

Metaphase chromosomes and interphase nuclei of chorionic villus samples (CVS) in five cases were studied after treatment with trypsin and post-fixation in formaldehyde by chromosomal in situ suppression (CISS) hybridization. Our modified protocol enables the use of in situ hybridization. techniques on CVS preparations after 42 h ofculture. A balanced translocation and trisomy 13 were identified with the aid of CISS hybridization.     

Chorionic villus culture for prenatal diagnosis of chromosome defects: Reduction of the long-term cultivation time

We report in detail two series of chorionic villus cultivation for prenatal chromosomal diagnosis. Chorionic villi were sampled from both first- and second-trimester pregnancies. One hundred cultures were treated with trypsin–EDTA for 2 h and collagenase overnight, (method A) and 100 were treated with trypsin–EDTA for 1 h and collagenase for 2 h (method B). Using short-term enzymatic digestion, the cultivation time was reduced from 14 days to 6 days. Sufficient amounts of metaphases of good quality were present in 93 per cent of primary cultures harvested in situ, whereas enough metaphases of sufficiently good quality were in most cases present only after subcultivation of the cultures using method A. The decrease in cultivation time obtained is probably due to a higher yield of viable cells in monocellular suspension, an increased attachment efficiency, and a more rapid attachment of single cells (within 24 h).     

Short communication kazal type trypsin inhibitor in amniotic fluid in fetal developmental disorders

The amniotic fluid concentrations of the Kazal type trypsin inhibitor were studied in pregnancies with fetal developmental disorders. The samples were obtained by amniocentesis between 14 and 19 weeks of gestation. In cases with fetal malformations, the level was below the normal 10th centile in 15 out of 28 cases (54 per cent, P<0.05) and above the normal 90th centile in 2 cases (7.1 per cent). Low values were common in cases with intrauterine fetal death or congenital nephrosis. The levels were normal in fetal chromosomal aberrations.     

Effects of protein properties on ultrafiltration membrane fouling performance in water treatment

Protein-like substances always induce severe ultrafiltration (UF) membrane fouling. To systematically understand the effect of proteins, regenerated cellulose UF membrane (commonly used for protein separation) performance was investigated in the presence of bovine serum albumin (BSA) under various water conditions. Results showed that although trypsin enhanced the membrane flux via proteolysis, catalysis took a long time. Membrane fouling was alleviated at high solution pH and low water temperature owing to the strong electrostatic repulsion force among BSA molecules. Both Na⁺ and Ca²⁺ could increase membrane flux. However, Ca²⁺ played a bridging role between adjacent BSA molecules, whereas membrane fouling was alleviated via a hydration repulsion force with Na⁺. The order of influence on membrane fouling was as follows: Ca²⁺ concentration > Na⁺ concentration > pH > temperature > trypsin concentration. Furthermore, a polyvinylidene fluoride UF membrane experiment showed that Ca²⁺ could reduce the fouling induced by BSA. Thus, the differences in UF membrane performance will have application potential for alleviating UF membrane fouling induced by proteins during water treatment.     

国产防锈水对威斯塔大鼠致癌性研究

对使用国产防锈水(主要成分为亚硝酸钠和三乙醇胺)的一个工厂进行肿瘤流行病学调查,发现该厂人员恶性肿瘤发病率略有增加。经长期给大白鼠饮用防锈水实验证明,该法可导致大鼠胰腺腺泡增生和不典型增生并能诱发出对照动物所没有的胰腺癌,同时还可引起胃粘膜的增生和胃乳头状瘤。     

An improved method of direct chromosome preparation from chorionic villus and high resolution banding technique

The trophoblast was dissociated from the underlying mesenchymal layer either with acetic acid after short-term prefixation or with mechanical power after fixation twice. The colcemid treatment time was shorted to 16 min and trypsin solution of low pH (6.2) was used for banding. By these steps, the quality of chromosome banding was greatly improved and complete standard chromosome diagnoses were made in 24 of 24 cases. With the modified technique, high resolution banding chromosomes were consistently obtained after short-term incubation.     

The distinction between arylsulphatases in chorionic villi

The relatively high activity of arylsulphatase C (ASC) in the placenta is a potential risk for the misdiagnosis of arylsulphatase A (ASA) or arylsulphatase B (ASB) deficiency in chorionic villus sampling when assayed by synthetic substrates. A clear distinction between these enzymes can be achieved in either the direct villi or the cultured villi cells. Interestingly, the activity of ASC differed significantly in cultured villi cells when prepared by two different methods, namely, minced villi versus treatment with trypsin and collagenase, while ASA and ASB were not affected by these treatments. Whether ASC was directly affected by one of these treatments or whether a selection of cells with different ASC levels was achieved is not yet clear, but this phenomenon clearly indicates the importance of precise definition of CVS preparations to correlate with the enzyme activity data.     

Biochemical analysis of meconium in fetuses presumed to have cystic fibrosis

A series of proteins (albumin, transferrin, a₁- antitrypsin, alpha-fetoprotein and pancreatic oncofetal antigen) and enzymes (γ-glutamyltranspeptidase, aminopeptidase M, alkaline phosphatase, a-glucosidase and protease) was measured in fetal meconium extracts. There were 19 fetuses thought to have cystic fibrosis (CF), 13 with neural tube defects, three with chromosome abnormalities and 19 normal controls, all with gestational ages between 18 and 21 weeks. With the exception of alpha-fetoprotein, all the proteins and enzymes were significantly elevated in the CF meconium extracts. The most definitive indicator of a CF fetus was the albumin concentration, where the mean level was five times that found in the control groups. However, five of 19 fetuses assumed to have CF had albumin in the normal range. In these cases the meconium protease levels were grossly elevated. Furthermore, in the same five fetuses meconium concentration of pancreatic oncofetal antigen, a protein synthesized in the fetal pancreas, was also greatly raised. We suggest that post-mortem examination of a fetus thought to have CF should include measurement of meconium albumin, protease and pancreatic oncofetal antigen.     

First-trimester prenatal diagnosis of cystic fibrosis using fibroblasts from a deceased index child to establish haplotypes

First-trimester prenatal diagnosis of cystic fibrosis (CF) using linked DNA markers is usually only possible if there is an index affected child to establish the haplotype of the parental chromosomes. We describe a prenatal diagnosis where fibroblasts, cultured from the skin of a deceased affected child and then held in frozen storage for 3 years, were used as the starting point for tracking the CF gene. The fetus was diagnosed as a homozygous normal and the diagnosis confirmed by immunoreactive trypsin testing after birth. It was also possible to establish heterozygosity in the aunt of the affected child.     

生物破乳菌XH-1的破乳有效成分研究

对破乳菌XH-1（Bacillus mojavensis）的破乳效能进行研究,并对该菌所产生的破乳剂有效成分及其理化性质进行考察。结果表明,破乳菌XH-1全培养液针对O／w型模型乳状液24h破乳率为86．0％,48h破乳率可达100％;XH-1菌的破乳有效成分为胞外分泌物质,该物质耐低温、不耐高温、经胃蛋白酶、胰蛋白酶及尿素处理后其基本失活;采用不同蛋白质粗体方法对破乳活性物质进行处理,发现粗提产物均具有较好的破乳功能,说明该物质具有蛋白质的特性;经可见-紫外分光光度法扫描及红外光谱扫描分析发现该生物破乳剂的有效成分为破乳菌株XH-1发酵产生的胞外蛋白类物质。     

Validation of a conductometric bienzyme biosensor for the detection of proteins as marker of organic matter in river samples

This article describes a conductometric bi-layer based bienzyme biosensor for the detection of proteins as a marker of organic matter in rivers. Proteins were chosen to be used as indicators of urban pollution. The working mechanism of the bienzyme biosensor is based on the enzymatic hydrolysis of proteins into several fractions (peptides and amino acids), which results in a local conductivity change depending of the concentration of proteins. In this work, we began with the optimization of biosensor response using bovine serum albumin (BSA) as standard protein. For this objective seven enzymatic biosensors were prepared: four enzymatic sensors with only one layer of enzyme (proteinase K, trypsin, pronase or protease X) and three other enzymatic sensors with two layers (first layer: membrane containing proteinase K; second layer: one of the three other enzymes: trypsin, pronase or protease X). The biosensors were obtained through the deposition of enzymatic layers and the cross-linking process between enzymes and BSA in saturated glutaraldehyde vapour. The response of the various biosensors, described previously, were compared with the values of total organic carbon (TOC), and those of organic nitrogen (Norg), as determined by the laboratory accredits (CEMAGREF of Lyon) using the traditional method of analysis (NF EN 1484, infrared spectroscopy) and (NF EN 25663, mineralization/colorimetry assay) respectively for each water sample obtained from di erent sites in Lyon (France). The linear correlations obtained with the response of the seven biosensors showed the most important indices of correlations for the biosensor with two enzymatic layers: proteinase K + pronase (pkp). The optimum conditions for the preparation of the pkp biosensor increased the sensitivity and gave a limit of quantification of 0.583 g/L for TOC and 0.218 g/L for Norg in water samples. This sensor shows good reproducibility (2.28%), a capacity to be used at temperatures range 10– 30°C (depending on the season) and moreover a long lifetime (5 weeks).     

Pancreatic and sacral agenesis in association with maternal diabetes mellitus: Case report

Sacral agenesis was diagnosed sonographically in the fetus of a diabetic mother and pancreatic agenesis was found additionally at autopsy.     

添加剂PEG对共混超滤膜结构和性能的影响

以不同分子量聚乙二醇 (PEG)作为PSf/A共混超滤膜的添加剂 ,测定所制备膜的水通量、截留率 .结果表明 ,膜的水通量随PEG分子量的变化呈现非线性关系 .当PEG分子量为4000时 ,水通量达到最大值115.2mL·(cm²·h)^-1,是不加PEG时的6倍 .通过超滤实验结果以及膜结构的电镜照片的观察 ,研究了PSf/A共混超滤膜体系中PEG作用的规律 .PEG分子量的增加改变了PSf/A铸膜液体系的相平衡关系 ,使铸膜液成为热力学不稳定体系 ,加速了相分离的发生 ;另外 ,PEG分子量的增加提高了铸膜液的粘度 ,引起相分离的延迟,这2种作用影响了膜的最终结构和性能.     

硅烷化粉煤灰固定化胰脂肪酶

以硅烷化粉煤灰为载体 ,戊二醛为交联剂 ,交联法固定化自制胰脂肪酶 ,考察了固定化时间、给酶量、戊二醛浓度对固定化酶活力的影响 ,并着重研究了固定化胰脂肪酶的热稳定性。固定化酶在 55℃仍可保持较高的酶活。     

Potential neoplastic effects of parathion-methyl on rat liver

The mutagenic and carcinogenic effects of parathion-methyl were examined by bacterial reverse assay and a long term experiment with Wistar rats. The potential mutagenic effect of parathion-methyl in Salmonella typhimurium TA100 bacterial cells was observed without rat liver S9 metabolic activation. Parathion-methyl was further investigated for pathological changes in rat pancreas and liver. The long-term rat experiments showed that parathion-methyl exposure for 3 months can cause pathological changes in rat pancreases acinar cells and pancreatic hepatocytes. Atypical acinar cell focuses (AACF) were determined in the liver and pancreas of the rats. The results from short-term Ames test and long-term rat experiments suggest that parathion-methyl would be potential carcinogenic.     

Potential neoplastic e ects of parathion-methyl on rat liver

The mutagenic and carcinogenic e ects of parathion-methyl were examined by bacterial reverse mutation assay and a long-term experiment with wistar rats. The potential mutagenic e ect of parathion-methyl in Salmonella typhimurium TA100 bacterial cells was observed without rat liver S9 metabolic activation. Parathion-methyl was further investigated for pathological changes in rat pancreas and liver. The long-term rat experiments showed that parathion-methyl exposure for 3 months can cause pathological changes in rat pancreases acinar cells and pancreatic hepatocytes. Atypical acinar cell focuses (AACF) were determined in the liver and pancreas of the rats. The results from short-term Ames test and long-term rat experiments suggested that parathion-methyl would be potential carcinogenic.     

In vitro characteristics of human fetal cells obtained from chorionic villus sampling and amniocentesis

In vitro characteristics of human fetal cells have been investigated after chorionic villus sampling at the first trimester and amniocentesis at the second trimester of pregnancy. Light microscopy revealed heterogeneous morphology of cell types in both the chorionic villus culture and the amniotic fluid cultures. Based on the experiments performed, chorionic villus cells are more sensitive to pronase, trypsin, and versene during subculture and have a higher DNA content per single cell and release more [¹²⁵I]-Beta-human chorionic gonadotropin into culture medium than those found in amniotic fluid cells. The practical applications of this study are discussed.     

酶法水解提取剩余活性污泥蛋白质条件实验

以凯式定氮法为测试方法,对6种蛋白酶在各自最优条件下水解剩余活性污泥蛋白质,进行污泥提蛋白实验。实验结果表明,碱性蛋白酶水解效果最好,最佳水解条件是:pH值为8.0,温度55℃,加酶量为2%,反应时间4 h,液固比4:1。最佳工作条件下,污泥蛋白质提取率为52.5%。污泥蛋白提取液中重金属及有害金属含量低于国家饲料卫生标准。