棉铃虫乙酰胆碱酯酶的底物专一性和发育期变化(英)

通过对河北省邯郸地区和山东省冠县的棉铃虫(Helicoverpa armigera H(?)bner)乙酰胆碱酯酶(AChE)研究,结果表明,棉铃虫乙酰胆碱酯酶对乙酰硫代胆碱(ATCh)和乙酰-β-甲基硫代胆碱(MeTCh)的比活力以及米氏常数(K_(?))值随其发育阶段呈有规律的变化,AChE比活力在幼虫期呈现两个高峰,一个在三龄,另一个在化蛹前;在蛹期AChE比活力没有明显的变化,而且比活力值比较低;到成虫期第4天有一个明显的高峰,比活力值高于其它任何虫态。K_(?)和最大反应速度(V_(max))的变化趋势基本上与比活力一致。棉铃虫AChE的K_(?)和比活力随其生长发育阶段的周期性变化对于指导用有机磷和氨基甲酸酯类杀虫药剂的化学防治具有重要的意义。不同地点采集到的棉铃虫AChE对AICh和MeTCh水解的活化能有所不同,邯郸地区的棉铃虫AChE水解MeTCh的活化能,蛹和成虫是幼虫期的3.9-4.3倍,而冠县棉铃虫水解MeTCh的活化能则变化不大。AChE水解ATCh的活化能,邯郸地区棉铃虫的AChE不同虫态间变化不大,冠县棉铃虫AChE,幼虫和成虫期约是蛹期的4倍。这说明不同生长发育时期,棉铃虫AChE对底物的水解所消耗的能量是不同的。棉铃虫幼虫AChE的最适反应条件是酶量以重量计为50-100mg,反应时间为10-20min,反应温度为35℃,反应体系的pH值为8.0。     

铽离子与RNA的荧光反应及RNA测定的研究

核糖核酸（ＲＮＡ）与稀土铽离子在ｐＨ５．０～６．５条件下能形成具有较强荧光的络合物,其激发峰在２８８ｎｍ处,发射峰为４９４ｎｍ及５４５ｎｍ处．ＲＮＡ在０．１～１０ｍｇ／Ｌ范围内与其荧光强度呈线性关系,其检出限为６．０×１０￣（－８）ｍｏｌ／Ｌ．腺苷酸,尿苷酸,胞苷酸对ＲＮＡ的干扰比较小,因此可在其存在下选择性地测定ＲＮＡ．     

Genomic organization and evolution of the soybean SB92 satellite sequence

Repetitive DNA sequences comprise a large percentage of plant genomes, and their characterization provides information about both species and genome evolution. We have isolated a recombinant clone containing a highly repeated DNA element (SB92) that is homologous to ca. 0.9% of the soybean genome or about 10⁵ copies. This repeated sequence is tandemly arranged and is found in four or five major genomic locations. FISH analysis of metaphase chromosomes suggests that two of these locations are centromeric. We have determined the sequence of two cloned repeats and performed genomic sequencing to obtain a consensus sequence. The consensus repeat size was 92 bp and exhibited an average of 10% nucleotide substitution relative to the two cloned repeats. This high level of sequence diversity suggests an ancient origin but is inconsistent with the limited phylogenetic distribution of SB92, which is found an high copy number only in the annual soybeans. It therefore seems likely that this sequence is undergoing very rapid evolution.     

Crystallization and preliminary crystallographic analysis of recombinant human P38 MAP kinase.

The recombinant human p38 MAP kinase has been expressed and purified from both Escherichia coli and SF9 cells, and has been crystallized in two forms by the hanging drop vapor diffusion method using PEG as precipitant. Both crystal forms belong to space group P2(1)2(1)2(1). The cell parameters for crystal form 1 are a = 65.2 A, b = 74.6 A and c = 78.1 A. Those for crystal form 2 are a = 58.3 A, b = 68.3 A and c = 87.9 A. Diffraction data to 2.0 A resolution have been collected on both forms.     

Identification of the Sites for Suppressor Mutations on the Hemagglutinin Molecule to Temperature-Sensitive Phenotype of the Influenza Virus

A temperature-sensitive (ts) mutant of the influenza virus A/WSN/ 33 strain, ts-134, possessed a defect in intracellular transport at the nonpermissive temperature and marked thermolability of hemagglutinin (HA) activity at 51 C. These were caused by a change at amino acid residue 157 from tyrosine to histidine in the HA protein. We isolated 37 spontaneous revertant clones from ts-134 at the nonpermissive temperature and determined their HA sequences. The deduced amino acid sequences demonstrated that one was a true revertant and the others were revertants with suppressor mutations, each of which had an additional amino acid change besides those of ts-134. The changed amino acids were located at 14 positions on the HA molecule, and eight of them were found in multiple revertants. These were located in five to six distinct regions on the three-dimensional structure of the HA molecule. However, the heat stability of HAs in the revertants was recovered differently depending on the sites of the changed amino acids. The kinetics of transport of the HA protein in the revertants were slightly delayed compared to the wild-type both at permissive and nonpermissive temperatures.     

Characterization and immunocytochemical demonstration of glucocorticoid receptor using antisera specific to transformed receptor

Cytosolic and nuclear forms of the glucocorticoid receptor were characterized using immunochemical techniques. Antibodies were raised in rabbits to an M_r 58,000 fragment of the transformed (DNA-binding) glucocorticoid receptor purified from rat liver cytosol by DNA-cellulose chromatography and polyacrylamide gel electrophoresis. Antibodies reacted with the transformed receptor form in a radioimmunoassay for glucocorticoid receptor. Western blot analysis of antibody reactivity revealed a single M_r 185,000 receptor form in rat liver cytosol but a smaller M_r 85,000 form in nucleosol, indicating the M_r 85,000 form is the transformed receptor. Furthermore, western blot analysis indicates that the M_r 185,000 receptor undergoes proteolysis during receptor purification and in vitro transformation processes by generating immunochemically similar proteins of smaller molecular weights. An identical M_r 185,000 glucocorticoid receptor was detected in cytosols of four rat tissues; liver, brain, adrenal medulla, and thymus. The glucocorticoid receptor was localized to the cytoplasm and nucleus of rat adrenal medulla cells by immunohistochemistry, demonstrating the existence in vivo of the transformed receptor and translocation of the receptor from cytoplasm to nucleus.     

Structural study of circulating thymic factor: a peptide isolated from pig serum. I. Isolation and purification.

A circulating thymic factor (FTS) has been characterized by a bioassay based on its ability to render theta-negative rosette-forming cells theta-positive and azathioprine-sensitive. FTS was sequentially purified and finally isolated from 1000 liters of pig serum by ultrafiltration, gel filtration, and ion exchange chromatography. Its amino acid composition and apparent molecular weight estimated by Sephadex G-25 chromatography, indicate that FTS is a nonapeptide of composition lysine, aspartic acid (or asparagine), serine 2, glutamic acid (or glutamine) 2, glycine 2, and alanine.     

Pindolol acts as beta-adrenoceptor agonist in orthostatic hypotension: therapeutic implications.

Three bedridden patients with severe orthostatic hypotension due to chronic autonomic failure were treated with pindolol (15 mg/day), a beta-adrenoceptor antagonist with partial agonist activity. While taking this drug the patients were free of orthostatic symptoms: they could walk, and standing blood pressure was maintained above 90/50 mm Hg. Supine heart rate rose during treatment by 12-21 beats/minute, and stroke volume and cardiac output by 12-24 ml and 1.5-3.1 l/min respectively. Supine blood pressure rose by 21-68 mm Hg systolic and 14-49 mm Hg diastolic. Pindolol 15 mg/day was therapeutically effective in these three patients with severe orthostatic hypotension due to chronic autonomic failure. Further studies in a larger series of patients are needed to confirm this result.