Ovarian superstimulatory response relative to follicular wave emergence in heifers

Two experiments were designed to evaluate the responsiveness of beef heifers to superstimulatory treatments administered during the first follicular wave. Heifers were examined daily (Experiment 1) or twice daily (Experiment 2) by ultrasonography to determine the status of follicular wave development and the day of initiation of superstimulatory treatment. Heifers in both experiments were superstimulated with a total dose of 10 ml Folltropin (equivalent to 200 mg of NIH-FSH-P1), divided into 10 equal intramuscular injections over 5 days. On the last day of treatment, heifers received 500 mug of cloprostenol after each injection of Folltropin to induce luteolysis. In the respective groups, superstimulatory treatments were initiated on Day -1, Day 0 (day of ovulation) or Day +1 for Experiment 1, and on Day -1, Day 0, Day +1 or Day +2 for Experiment 2. In Experiment 1, the number of ovulations in each ovary was assessed by ultrasonography and by counting the number of corpora lutea (CL) in each ovary at slaughter. The correlation between both techniques for assessing ovulatory response was high (r= 0.98; P< 0.0001), and there was no significant difference in the mean number of ovulations detected by ultrasound (5.7+/-1.1) versus the mean number of CL counted at slaughter (6.2+/-1.2). In Experiment 1, the mean (+/- SEM) number of CL counted at slaughter in heifers treated on Day -1 (9.4+/-3.8) and Day 0 (7.3+/-1.6) was higher (P< 0.05) than that of heifers treated on Day +1 (0.7+/-0.3). The mean number of follicles >/=7 mm in diameter on the last day of treatment was also higher (P<0.05) in the Day -1 group compared with the Day +1 group; the Day 0 group was intermediate. In Experiment 2, the mean number of ovulations was higher (P< 0.05) in the Day 0 group (18.4+/-3.4) than the Day -1 (9.5+/-2.3), Day +1 (6.7+/-2.2) or Day +2 (6.5+/-2.3) groups. Heifers in the Day -1, and Day 0 groups had more (P< 0.05) follicles >/=7 mm at the end of treatment compared with heifers in the Day +1 or the Day +2 group. The stated hypothesis was supported: exogenous FSH treatment initiated at the time of wave emergence, near the expected time of the endogenous wave-eliciting FSH surge, has a positive effect on the superstimulatory response. A higher superstimulatory response was elicited when treatments were initiated on the day of, or the day before, wave emergence compared with that of later treatments.     

Rat liver foci study on coexposure with 50 Hz magnetic fields and known carcinogens

A study was performed to investigate possible interactions by magnetic fields (MF) with the processes of initiation and promotion of chemically induced preneoplastic lesions in rat liver. Male Sprague-Dawley rats were subjected to a 70% partial hepatectomy followed after 24 h by i.p. injection of diethylnitrosamine (DENA) as a tumour initiator. Starting one week after the DENA-treatment phenobarbital (PB) was given to promote growth of enzymatically altered foci of liver cells. MF was applied immediately after the partial hepatectomy and continued until sacrifice after 12 weeks of PB exposure. Homogenous horizontal AC magnetic fields with a frequency of 50 Hz and flux densities of 0.5 μT or 0.5 mT were used. The rats coexposed with MF and DENA plus PB did not gain weight as much as the rats exposed to the chemical agents only. The MF-exposure also resulted in a slight reduction in size and numbers of the focal lesions. The results suggest an interaction of MF with the processes of chemical carcinogenesis either as a result of stress or depending on effects on the proliferation of preneoplastic cells. © 1993 Wiley-Liss, Inc.     

重组人IL6／2融合蛋白对6.5Gy照射小鼠造血功能恢复的影响

观察了ｈＦＰＩＬ６／２对６．５Ｇｙγ线照射ＮＩＨ小鼠第１０天造血功能恢复的影响。结果表明：照射小鼠连续４ｄ给予ｈＦＰＩＬ６／２２５０μｇ·ｋｇ－１·ｄ－１,其脾重、ＣＦＵ－８、骨髓有核细胞数及ＣＭ－ＣＦＵ分别比对照组增加５９．０％、２７８．５％、５７．９％和１３８．２％,统计学处理均有显著差异;对此四项指标的改善也明显优于２５μｇ组。另外,２５０μｇ剂量组小鼠外周血象３０ｄ的动态观察结果表明,ｈＦＰＩＬ６／２不但能明显提高红细胞和血红蛋白的最低值,而且能使血小板的恢复提前。提示ｈＦＰＩＬ６／２在促进血小板生成和促进红系造血方面可能具有良好的应用前景。     

内皮素对失血性休克鼠脑微循环的影响及尼莫地平、川芎嗪的治疗作用

利用大鼠颅骨开窗观察软脑膜微循环的方法研究了内皮素（ＥＴ－１）１０－１０－１０－７ｍｏｌ／Ｌ对软脑膜微循环的影响以及失血性休克时软脑膜对ＥＴ－１的反应性。并用１０－７ｍｏｌ／Ｌ造成失血性休克后脑血管痉挛的模型,观察尼莫地平、川芎嗪、６５４－２对内皮素引起血管痉挛的治疗作用。１０－９、１０－８和１０－７ｍｏｌ／Ｌ３种浓度ＥＴ－１可使软脑膜小动脉、细动脉强烈收缩,收缩率分别为２７．７％、４６．８％、７８．５％,其收缩强度与ＥＴ－１的浓度有关。对静脉的作用不明显。１０－１０ｍｏｌ／ＬＥＴ－１可使细动脉轻度扩张。出血性休克时,软脑膜血流明显减慢,小动脉、细动脉管径对ＥＴ－１的收缩作用更敏感,脑组织血流明显减少。尼莫地平具有较好的拮抗ＥＴ－１引起软脑膜动脉的收缩和改善局部微循环的作用。川芎嗪也能拮抗ＥＴ－１引起软脑膜动脉的收缩,但作用较尼莫地平弱。６５４－２不能缓解ＥＴ－１对软脑膜动脉的收缩作用。     

慢性缺氧对大鼠膈肌组织化学和超微结构影响的研究

研究慢性缺氧和参麦注射液治疗对大鼠膈肌组织化学和超微结构的影响,结果显示：缺氧使膈肌ＳＤＨ活性降低,Ⅰ类纤维极显著减少,运动终板ＣｈＥ活性极显著降低,线粒体肿胀变性,神经肌肉运动终板突触前部囊泡减少,突触间隙模糊,终板模电子密度降低;参麦治疗可恢复缺氧膈肌的ＳＤＨ活性及Ⅰ类纤维数目,显著提高终板ＣｈＥ活性,线粒体结构恢复正常,运动终板突触前部囊泡丰富,突触间隙清晰,终板膜电子密度正常。表明慢性缺氧降低膈肌有氧氧化能力,影响其能量代谢及神经冲动的传递,从而减弱其收缩力,而参安治疗可改善缺氧造成的上述损害,为其临床应用提供了实验医学依据。     

Na+, K+, Cl− cotransport and its regulation in Ehrlich ascites Tumor cells. Ca2+/Calmodulin and protein kinase C dependent pathways

Summary Net Cl^– uptake as well as unidirectional³⁶Cl influx during regulatory volume increase (RVI) require external K⁺. Half-maximal rate of bumetanide-sensitive³⁶Cl uptake is attained at about 3.3mm external K⁺. The bumetanide-sensitive K⁺ influx found during RVI is strongly dependent on both Na⁺ and Cl^–. The bumetanide-sensitive unidirectional Na⁺ influx during RVI is dependent on K⁺ as well as on Cl^–. The cotransporter activated during RVI in Ehrlich cells, therefore, seems to transport Na⁺, K⁺ and Cl^–. In the presence of ouabain and Ba⁺ the stoichiometry of the bumetanide-sensitive net fluxes can be measured at 1.0 Na⁺, 0.8 K⁺, 2.0 Cl^– or approximately 1 : Na, 1 : K, 2 : Cl. Under these circumstances the K⁺ and Cl^– flux ratios (influx/efflux) for the bumetanide-sensitive component were estimated at 1.34 ±0.08 and 1.82 ± 0.15 which should be compared to the gradient for the Na⁺, K⁺, 2Cl^– cotransport system at 1.75 ± 0.24.Addition of sucrose to hypertonicity causes the Ehrlich cells to shrink with no signs of RVI, whereas shrinkage with hypertonic standard medium (all extracellular ion concentrations increased) results in a RVI response towards the original cell volume. Under both conditions a bumetanide-sensitive unidirectional K⁺ influx is activated. During hypotonic conditions a small bumetanide-sensitive K⁺ influx is observed, indicating that the cotransport system is already activated.The cotransport is activated 10–15 fold by bradykinin, an agonist which stimulates phospholipase C resulting in release of internal Ca²⁺ and activation of protein kinase C.The anti-calmodulin drug pimozide inhibits most of the bumetanide-sensitive K⁺ influx during RVI. The cotransporter can be activated by the phorbol ester TPA. These results indicate that the stimulation of the Na⁺, K⁺, Cl^– cotransport involves both Ca²⁺/calmodulin and protein kinase C.     

Coupling Among Energy Failure, Loss of Ion Homeostasis, and Phospholipase A2 and C Activation During Ischemia

Abstract— The objective of the present experiments was to correlate changes in cellular energy metabolism, dissipative ion fluxes, and lipolysis during the first 90 s of ischemia and, hence, to establish whether phospholipase A₂or phospholipase C is responsible for the early accumulation of phospholipid hydrolysis products. Ischemia was induced for 15–90 s in rats, extracellular K⁺ (K⁺_e) was recorded, and neocortex was frozen in situ for measurements of labile tissue metabolites, free fatty acids, and diacylglycerides. Ischemia of 15-and 30-s duration gave rise to a decrease in phosphocreatine concentration and a decline in the ATP/free ADP ratio. Although these changes were accompanied by an activation of K⁺ conductances, there were no changes in free fatty acids until after 60s, when free arachidonic acid accumulated. An increase in other free fatty acids and in total diacylglyceride content did not occur until after anoxic depolarization. The results demonstrate that the early functional changes, such as activation of K⁺ conductances, are unrelated to changes in lipids or lipid mediators. They furthermore suggest that the initial lipolysis occurs via both phospholipase A₂ and phospholipase C, which are activated when membrane depolarization leads to influx of calcium into cells.     

Bacterial Disproportionation of Elemental Sulfur Coupled to Chemical Reduction of Iron or Manganese

A new chemolithotrophic bacterial metabolism was discovered in anaerobic marine enrichment cultures. Cultures in defined medium with elemental sulfur (S⁰) and amorphous ferric hydroxide (FeOOH) as sole substrates showed intense formation of sulfate. Furthermore, precipitation of ferrous sulfide and pyrite was observed. The transformations were accompanied by growth of slightly curved, rod-shaped bacteria. The quantification of the products revealed that S⁰ was microbially disproportionated to sulfate and sulfide, as follows: 4S⁰ + 4H₂O → SO₄^2- + 3H₂S + 2H⁺. Subsequent chemical reactions between the formed sulfide and the added FeOOH led to the observed precipitation of iron sulfides. Sulfate and iron sulfides were also produced when FeOOH was replaced by FeCO₃. Further enrichment with manganese oxide, MnO₂, instead of FeOOH yielded stable cultures which formed sulfate during concomitant reduction of MnO₂ to Mn²⁺. Growth of small rod-shaped bacteria was observed. When incubated without MnO₂, the culture did not grow but produced small amounts of SO₄^2- and H₂S at a ratio of 1:3, indicating again a disproportionation of S⁰. The observed microbial disproportionation of S⁰ only proceeds significantly in the presence of sulfide-scavenging agents such as iron and manganese compounds. The population density of bacteria capable of S⁰ disproportionation in the presence of FeOOH or MnO₂ was high, > 10⁴ cm^-3 in coastal sediments. The metabolism offers an explanation for recent observations of anaerobic sulfide oxidation to sulfate in anoxic sediments.     

Reactions Catalyzed by Peg-Modified α-Chymotrypsin in Organic Solvents. Influence of Water Content and Degree of Modification

-Chymotrypsin was modified with cyanuric chloride activated monomethoxypolyethylene glycol (MPEG) with molecular weights 1900 and 5000. Using the higher molecular weight MPEG a product that was soluble in benzene at moderate levels of modification was obtained, whereas with MPEG 1900 almost all the enzyme's amino groups had to be modified for dissolving the conjugate. The catalytic activity decreased with increasing degree of substitution. Apparent V_max was considerably higher for the less modified enzyme preparation than for the more modified one, while K_m,app stayed almost constant. The modified enzyme was used for peptide synthesis. The reaction was dependent on the content of dissolved water. Both V_max,app and K_m,app increased with increasing water content. It was possible to achieve a process with complete conversion of substrate to dipeptide.