Microcarrier culture of bowes melanoma cells in serum-free medium with Human plasma fraction IV-4+ V

Bowes melanoma cells were cultivated successfully in a serum-free medium which was constructed by the concept of maximum retention of proteins from fractionated human plasma having growth stimulatory activities. The cells could be cultivated in the serum-free medium without any adaptation period. The major serum-free component of the medium was the fraction IV-4 + V of the Cohn fractionation process of human plasma. Approximately six times increase of tissue-type plasminogen activator (t-PA) activity as compared with that in serum-free medium even though the cell growth was much slower. In addition, the growth stimulatory activities of thrombin and fibronectin were investigated during the cultivation of Bowes melanoma cells in this serum-free medium. These proteins contributed significantly to the enhanced growth of cells by reducing doubling time to 25 and 35 h as compared with 55 h in the serum-free medium without them. Especially, fibronectin supported cells to propagate near to the maximum cell density achieved in the medium with 10% FBS.     

DDT inhibition of Ca-Mg ATPase from peripheral nerves and muscles of lobster, Homarus americanus

Sensitivity of CaMg ATPase from axonic plasma membrane (APM) and sarcoplasmic reticulum (SR) of lobster, $\text{Homarus}\text{,}\text{americanus}$, to DDT was studied. The CaMg ATPase found in SR with the high Ca²⁺ affinity is sensitive to DDT while the portion of ATPase related to the low Ca²⁺ affinity site is not inhibited by DDT. Also, DDT is more inhibitory against the CaMg ATPase prepared from APM than the one obtained from SR. The relationship between inhibition of the CaMg ATPase by DDT in the axonic nerve membrane and $\text{in}\text{,}\text{vivo}$ poisoning symptoms of the nervous system is discussed.     

Simultaneous tissue profiling of eicosanoid and endocannabinoid lipid families in a rat model of osteoarthritis

We describe a novel LC method for the simultaneous and quantitative profiling of 43 oxylipins including eicosanoids, endocannabinoids, and structurally related bioactive lipids with modified acyl groups. The LC-MS/MS method uses switching at a defined time between negative and positive electrospray ionization modes to achieve optimal detection sensitivity for all the lipids. The validated method is linear over a range of 0.01–5 nmol/g (0.1–50 nmol/g for 2-arachidonoyl glycerol) with intra- and interday precision and accuracy between 1.38 and 26.76% and 85.22 and 114.3%, respectively. The method successfully quantified bioactive lipids in different tissue types in the rat, including spinal cord, dorsal root ganglia (DRGs), knee joint, brain, and plasma. Distinct regional differences in the pattern of lipid measured between tissue types were observed using principle component analysis. The method was applied to analyze tissue samples from an established preclinical rat model of osteoarthritis (OA) pain and showed that levels of 12-hydroxyeicosatetraenoic acid were significantly increased in the OA rat knee joint compared with controls, and that 15-hydroxyeicosatetraenoic acid was significantly increased in the DRGs in the model of OA compared with controls. The developed LC-MS/MS method has the potential to provide detailed pathway profiling in tissues and biofluids where the disruption of bioactive oxylipins may be involved in disease states.     

秋冬季叶施N、Fe提高假俭草草坪抗寒性研究

在过渡性气候地区的假俭草草坪,宜在最低气温为12℃时(11月中、下旬)叶施Fe,最佳施用浓度为:单施Fe为0.6L/m²,N+Fe为4g/m² N+0.6L/m²Fe。秋冬季叶施Fe、N+Fe可提高草体内抗寒指标性物质的含量,叶施0.6L/m² Fe或4g/m² N+0.6L/m² Fe,可使草坪草体内脯氨酸、可溶性糖类含量及过氧化氢酶的活性分别提高25%、310%、180%和41%、182%、160%。叶施N+Fe处理的效果比单独施Fe的效果更好,施后草坪枯黄期比对照组推迟6~8d,半致死温度降低3~4℃。     

植物抗冷分子生物学研究进展（综述）

温度是植物生长的必要条件,然而低温却是限制作物生产的重要因素,为此,各国政府及研究部门一直都把植物低温适应性问题作为一个重要的研究课题．按照低温的不同程度,植物的低温伤害可分为冷害（ｃｈｉｌｌｉｎｇ ｉｎｊｕｒｙ;零上低温对植物的伤害）和冻害（ｆｒｅｅｚｉｎｇ ｉｎｊｕｒｙ;零下低温对植物的伤害）两大类。早期关于植物冷害机理和抗冷机理的研究,是从比较冷敏感（ｃｈｉｌｌｉｎｇ－ｓｅｎｓｉｔｉｖｅ）植物和抗冷（ｃｈｉｌｌｉｎｇ－ｉｎｓｅｎｓｉｔｉｖｅ）植物或比较未经冷驯化的植物（ｎｏｎ－ｃｏｌｄ－ａｃ…     

Electric discharge plasmas influence attachment of cultured CHO K1 cells

Non-thermal plasmas can be generated by electric discharges in gases. These plasmas are reactive media, capable of superficial treatment of various materials. A novel non-thermal atmospheric plasma source (plasma needle) has been developed and tested. Plasma appears at the end of a metal pin as a submillimetre glow. We investigate the possibility of applying the plasma needle directly to living tissues; the final goal is controlled cell treatment in microsurgery. To resolve plasma effects on cells, we study cultured Chinese hamster ovarian cells (CHO-K1) as a model system. When these are exposed to the plasma, instantaneous detachment of cells from the surface and loss of cell-cell interaction is observed. This occurs in the power range 0.1-0.2 W. Cell viability is assessed using propidium iodide (PI) and cell tracker green (CTG) fluorescent staining utilizing confocal laser scanning microscopy (CLSM). Detached cells remain alive. Use of higher doses (plasma power >0.2 W) results in cell necrosis. In all cases, plasma-influenced cells are strictly localized in submillimetre areas, while no reaction in surrounding cells is observed. Due to its extreme precision, plasma treatment may be applicable in refined tissue modification.     

Association mapping of cold‐induced sweetening in potato using historical phenotypic data

In this study, historical phenotypic data from a potato breeding programme were used with an association mapping approach to identify alleles of candidate genes associated with cold‐induced sweetening of potato. Molecular marker analysis was used to determine allelic variation of candidate genes potentially involved in cold‐induced sweetening. Variations in the UDP‐glucose pyrophosphorylase (UGPase, EC 2.7.7.9) and apoplastic invertase genes (EC 3.2.1.26) were significantly associated with cold‐induced sweetening, and a possible interaction of apoplastic invertase and apoplastic invertase inhibitor was identified. This demonstrates that breeding programme phenotypic data collected over multiple years and environments can be used successfully with pedigree information for association mapping. It also confirms that the UGPase and apoplastic invertase markers are transferable across breeding programmes with distinct germplasm.     

不同抗凝人血浆凝固酶检测的探讨

目的 了解3种不同抗凝人血浆检测血浆凝固酶的可行性及观察时间对结果的影响.方法 用3种不同抗凝人血浆及以不同菌液浓度作试管法凝固酶试验.结果 EDTA-K2抗凝血浆出现阳性慢,凝块大多较小,24 h内稳定性较好;枸橼酸钠抗凝血浆出现结果介于EDTA-K2、肝素抗凝血浆之间,结果最不稳定;肝素抗凝血浆出现阳性最快,但不够稳定;菌液浓度对出现阳性的早晚无影响.结论 3种抗凝人血浆在试管法凝固酶测定中使用时,出现阳性快慢不受菌液浓度影响,随时间延长溶解现象可能会增加,应掌握结果观察时间,以降低假阴性.     

The effect of assay composition, detergent solubilization and reconstitution on red beet (Beta vulgaris L.) plasma membrane H-ATPase kinetic properties

Modification of the salt concentration, composition and/or buffer type in the assay of plasma membrane ATPase activity caused substantial changes in the K_m and slight changes in the temperature dependence of this enzyme. The K_m and temperature dependence were also affected by detergent solubilization of the ATPase and its subsequent reconstitution into liposomes. Modulation of kinetic properties by assay composition and hydrophobic state reflect the sensitivity of the plasma membrane H⁺-ATPase to its immediate environment. This may indicate a possible regulatory mechanism for this important plant enzyme.     

Effects of postnatal treatment with naloxone on plasma gonadotropin,prolactin, testosterone and testicular functions in male rats

Opioid peptides are implicated in the control of gonadotropin and prolactin secretion. The role of opioid antagonist naloxone and its effects on plasma gonadotropin, prolactin, testosterone levels and testicular hyaluronidase, acid phosphatase, [³H]uridine and thymidine incorporation, RNA, DNA and protein concentrations were evaluated in rats after administration of naloxone beginning day 1 through 21 and autopsied on 45, 60 and 90 days of age. Plasma gonadotropin and testosterone levels were significantly elevated after naloxone treatment. Testicular hyaluronidase and acid phosphatase activity increased till 60 days post treatment and declined thereafter. Concentrations of RNA and protein did not change significantly but the concentration of DNA declined at 45 and 60 days of age. These results suggest that endogenous opioid peptides exert regulatory influence on gonadotropin secretion which in turn control the testicular function in the male rat.