A high affinity,calmodulin-responsive (Ca2+ + Mg2+)-ATPase in isolated bone cells

Although acute alterations in Ca²⁺ fluxes may mediate the skeletal responses to certain humoral agents, the processes subserving those fluxes are not well understood. We have sought evidence for Ca²⁺-dependent ATPase activity in isolated osteoblast-like cells maintained in primary culture. Two Ca²⁺-dependent ATPase components were found in a plasma membrane fraction: a high affinity component (half-saturation constant for Ca²⁺ of 280 nM, $\text{V}{}_{\mathrm{<mtext>max</mtext>}}$ of 13.5 nmol/mg per min) and a low affinity component, which was in reality a divalent cation ATPase, since Mg²⁺ could replace Ca²⁺ without loss of activity. The high affinity component exhibited a pH optimum of 7.2 and required Mg²⁺ for full activity. It was unaffected by potassium or sodium chloride, ouabain or sodium azide, but was inhibited by lanthanum and by the calmodulin antagonist trifluoperazine. This component was prevalent in a subcellular fraction which was also enriched in 5′-nucleotidase and adenylate cyclase activities, suggesting the plasma membrane as its principal location. Osteosarcoma cells, known to resemble osteoblasts in their biological characteristics and responses to bone-seeking hormones, contained similar ATPase activities. Inclusion of purified calmodulin in the assay system caused small non-reproducible increases in the Ca²⁺-dependent ATPase activity of EGTA-washed membranes. Marked, consistent calmodulin stimulation was demonstrated in membranes exposed previously to trifluoperazine and then washed in trifluoperazine-free buffer. These results indicate the presence of a high affinity, calmodulin-sensitive Ca²⁺-dependent ATPase in osteoblast-like bone cells. As one determinant of Ca²⁺ fluxes in bone cells, this enzyme may participate in the hormonal regulation of bone cell function.     

MICROSPECTROPHOTOMETRIC ANALYSIS OF NUCLEAR DNA IN CHARA ZEYLANICA

Microspectrophotometric analysis of the DNA content of nuclei in various parts of Chara zeylanica Willd. revealed that the amount of DNA in the nucleus of an internodal cell equals twice the amount of DNA in the nucleus of a sperm, while the half-anaphase stage of the same nodal cells contains the same amount of DNA as the nuclei of the male gametes. The DNA content of the nuclei of internodal cells may rise as much as 50 times higher than that of the gametes. However, in the oldest (most basal) internodal cells, the DNA content of the minute nuclei falls again to the basic (1 C) amount. Measurements of sister nuclei derived by amitosis indicated that both nuclei have equal amounts of DNA; this was interpreted as further evidence that amitosis is not a disorganized process or manifestation of degeneration. The bearing of these analyses on the question of the site of meiosis in these plants is discussed.     

Behaviour of photosynthetic products associated with growth and grain production in the rice plant

Summary A study of the behaviour of the photosynthetic products assimilated at different growth stages was conducted in the field and in the greenhouse using C¹⁴ tracer.In general, the assimilated carbon is translocated to and accumulates in the growing organs. The carbon assimilated at the maximum tiller number stage is distributed mostly to the lower leaves. The carbon assimilated at the booting stage is distributed mostly to the spikelet, certain leaf sheaths and culms. The carbon accumulated in the form of carbohydrates in the leaf sheaths and the culm before flowering is retranslocated to the panicle after flowering. However, because of the consumption by respiration, the efficiency of this type of carbohydrate in grain production is not very high. The carbon assimilated after flowering accumulated mostly and efficiently in the brown rice.The release of the assimilated carbon as CO₂ is most intense immediately after assimilation. Thirty-five to 60 per cent of the assimilated carbon is consumed through respiration under the conditions of this experiment. As the carbon, which is in the form of sugars, rapidly changes to other forms, and also is consumed by respiration, the consumption declines rapidly. The retention percentage of assimilated carbon decreases as mutual shading increases.The large proportion of carbon released through respiration indicates the importance of studies on the significance of respiration in relation to growth.A portion of the thesis for the Master of Science degree submitted by Mr. Shen Lian to the Graduate School, University of the Philippines, College of Agriculture.     

生物素标记寡核苷酸探针探测扩增的病理性突变珠蛋白基因

用末端转移酶催化生物素核苷酸底物(Biotin-ll-dUTP)共价连接在合成的寡核苷酸3’羟基末端,从而合成了两种寡核苷酸探针(β~T_(41-42)及β~A_(41-42))。用它们分别与克隆化扩增的正常和突变的β—珠蛋白基因片段杂变。结果表明该探针都具有与~(32)P探针相似的特异性,其杂交的灵敏度为2—3pg(特异序列)。进而将探测HbS基因的正常和异常两种寡核苷酸19聚体(β~A_6和β~S_6)用~(32)P和生物素分别标记;将HbS杂合子病人的白细胞DNA经聚合酶链反应(PCR)法扩增,并以含正常β—珠蛋白基因的DNA片段作对照,与两种探针分别进行斑点杂交。所得结果完全一致;Hbs杂合子DNA对正常和异常探针都显出杂交信号,而正常DNA只与β~A探针显杂交信号。     

三乙醇胺基强碱性聚苯乙烯树脂共价固定胰酶的研究

用多孔强碱型三乙醇胺基聚苯乙烯阴离子交换树脂做为载体,用CNBr与载体上的多羟基作用共价偶联了胰酶。红外光谱表明:其共价偶联反应机理与用CNBr活化多糖类载体并接酶的机理相类似。最适偶联条件研究表明:CNBr用量增多,酶蛋白载量增加。但比活下降。偶联pH为10时,固定化酶有适宜的载量和较高的比活。由于胰酶水解蛋白反应释放出H~+质子,这些质子在载体内积累,使微环境内H~+质子浓度增加,进而使得固定化胰酶的pH—活性曲线在pH9～11范围内未出现下降。在变温和60℃恒温下对固定化酶的热稳定性测试表明:固相酶的热稳定性比天然酶的热稳定性有所提高。     

Stereoselective high-performance liquid chromatographic assay for propranolol enantiomers in serum

A simple and sensitive stereoselective high-performance liquid chromatographic assay for the quantitation of propranolol enantiomers in serum is described. The method involves conversion of the propranolol enantiomers to diastereomeric urea derivatives by reaction with the chiral reagent (+)-phenylethylisocyanate, followed by chromatographic separation of the diastereomeric products. Conditions of the derivatization reaction were optimized to achieve rapid and quantitative yield with either of the enantiomers. Baseline resolution of the diastereomers was achieved on a reversed phase C₈ column with an isocratic mobile phase. Fluorescence detection afforded an absolute on-column detection limit of 100 pg. The assay has been applied to pharmacokinetic studies in humans and small laboratory animals.     

厚孢孔菌属一新种

本文报道了厚孢孔菌属(Pachykytospora)的一个新种,即大厚孢孔菌Pachykytospora major G.Y.Zheng et Bi和一个国内新记录:瘤厚孢孔菌Pachykytospora tuberculosa(DC.:Fr.)Kotl.et Pouz.。     

PUC衍生质粒的一个低拷贝数突变型

本文报道大肠杆菌的ColE1类似质粒的一个低拷贝数突变型。从载体质粒pUC4衍生的重组质粒pPGVT3在大肠杆菌宿主DF2145中是不稳定的,以pPGVT3转化DF2145时在4o℃培养得不到转化子。用诱发点突变的羟胺体外处理pPGVF3质粒DNA,得到一个稳定性提高了的突变质粒pPGVT3HA,突变的位置被确定在质粒的pUC部分,突变降低了pUC及其衍生质粒的拷贝数。文中对质粒的稳定性与拷贝数的关系作了讨论。     

超干处理对几种芸苔属植物种子生理生化和细胞超微结构的效应

几种芸苔属植物种子经过超干处理后,发芽力和活力不受影响,耐藏性大为提高。超干及老化后种子的脱氢酶、超氧物歧化酶、过氧化物酶、过氧化氢酶等酶系统保持完好;丙二醛和挥发性醛类物质等劣变产物相应减少;电导率测定表明,细胞膜系统的完整性良好;透射电镜(TEM)和扫描电镜(SEM)观察表明,细胞超微结构及功能保持完好。不同干燥方法和不同干燥剂的系列实验证实,生石灰的脱水效果并不亚于冰冻真空干燥法,且经济实用。     

聚乙二醇处理大豆种子子叶中几种酶活性和可溶性蛋白含量的变化

聚乙二醇处理的大豆种子的异柠檬酸裂解酶、苹果酸脱氢酶、过氧化氢酶、超氧物歧化酶、酸性磷酸酶、碱性磷酸酶的活性明显高于受低温吸胀冷害的种子子叶的活性,相关的酶活性协同地增长,而蛋白质的含量没有明显的变化。这些酶活性的提高可能是渗透调控处理对细胞膜系统修补的结果。