Inverse agonists and neutral antagonists at µ opioid receptor (MOR): possible role of basal receptor signaling in narcotic dependence

The mu opioid receptor, MOR, displays spontaneous agonist-independent (basal) G protein coupling in vitro. To determine whether basal MOR signaling contributes to narcotic dependence, antagonists were tested for intrinsic effects on basal MOR signaling in vitro and in vivo, before and after morphine pretreatment. Intrinsic effects of MOR ligands were tested by measuring GTPgammaS binding to cell membranes and cAMP levels in intact cells. beta-CNA, C-CAM, BNTX, and nalmefene were identified as inverse agonists (suppressing basal MOR signaling). Naloxone and naltrexone were neutral antagonists (not affecting basal signaling) in untreated cells, whereas inverse agonistic effects became apparent only after morphine pretreatment. In contrast, 6alpha- and 6beta-naltrexol and -naloxol, and 6beta-naltrexamine were neutral antagonists regardless of morphine pretreatment. In an acute and chronic mouse model of morphine-induced dependence, 6beta-naltrexol caused significantly reduced withdrawal jumping compared to naloxone and naltrexone, at doses effective in blocking morphine antinociception. This supports the hypothesis that naloxone-induced withdrawal symptoms result at least in part from suppression of basal signaling activity of MOR in morphine-dependent animals. Neutral antagonists have promise in treatment of narcotic addiction.     

Topology of the 10 subunits within the Decamer of KLH, the hemocyanin of the marine gastropod Megathura crenulata

Immunoelectron microscopy has been performed using negatively stained immune complexes of keyhole limpet hemocyanin isoform 1 (KLH1) decamers and a functional unit-specific monoclonal antibody anti-KLH1-c1. The antibody links hemocyanin molecules at both the collar and the collarless edge of the decamer, indicating a peripheral localization of functional units c. In isoform 2 (KLH2) the positions of functional units c have been identified with the peanut agglutinin (PNA), which has previously been shown to exclusively bind to KLH2-c. Ferritin linked to PNA was used to visualize labeled molecules electron microscopically. The pattern of labeling also indicates a peripheral localization of the c functional units. The data presented in this paper support only one of two possible models for the subunit orientation within the hemocyanin decamer.     

Recovery and purification of aprotinin from industrial insulin-processing effluent by immobilized chymotrypsin and negative IMAC chromatographies

Aprotinin, a bovine protease inhibitor currently also produced in recombinant bacteria, yeast, and corn, has valuable applications as a human therapeutic and in tissue culture. The objective of this work was to develop the basis of a large-scale aprotinin purification process centered on immobilized metal ion affinity chromatography (IMAC). This technique uses ligands—metal ions—of a lower cost and higher stability than those traditionally used in affinity chromatography. Since aprotinin does not interact with IMAC ligands, collection is from the nonretained fractions (negative chromatography). Stirred-tank batch IMAC adsorption experiments indicated that one-step aprotinin purification could not be successful. Immobilized chymotrypsin chromatography was then used as a prepurification step, yielding a suitable feed for IMAC (with purification factors as high as 476). IMAC column fed with these prepurified materials produced purified aprotinin in the nonretained fractions with purification factors as high as 952.     

球形红假单胞菌反应中心中蛋白的量子化学研究

采用重叠二体近似方法和建立在从算水平上的扩展负本征值因数计算方法（extended negative factor counter method）研究了球形红假单胞菌（Rhodobacter sphaeroides(Van Niel)Imhoff,Truper et Pfennin）中兴合反应中心中蛋白链L及M的电子结构。结果表明：（1）对组成蛋白链L（M）的前线轨道有重要贡献的氨基酸残基分布在L链的自由螺旋区（M链的α螺旋区）。由于自由螺旋是有柔曲性的,它易于在电子转移的过程中改变其构象并降低体系的能量,而α螺旋结构却相对稳定,这种差别有可能是光合反庆中心中电子转移只沿L支进行的原因之一。（2）与特殊对分子及辅助叶绿素分子形成轴向配位的组氨酸残基对于特殊对P和辅助叶绿素分子的ELUMO有重要影响,但此组氨酸的相应分子轨道的贡献并没有出现在蛋白链的前线轨道组成中。这意味着色素分子与蛋白链之间的相互作用对蛋白链前线轨道的贡献没有影响,但却能影响相应色素分子的ELUMO能级。     

Computational approaches to motor control

New concepts and computational models that integrate behavioral and neurophysiological observations have addressed several of the most fundamental long-standing problems in motor control. These problems include the selection of particular trajectories among the large number of possibilities, the solution of inverse kinematics and dynamics problems, motor adaptation and the learning of sequential behaviors.     

Toxoplasma gondii: Electron microscopic study on the dye test reaction

When intact Toxoplasma trophozoites were stained with isotonic alkaline methylene blue solution, the organelles rich in nucleic acid, i.e., nucleus, free and membrane-bound ribosomes appeared as electron dense areas. When the parasites were incubated with the anti-Toxoplasma antibody and the accessory factor, swelling of the surface membrane occurred first, followed by destruction of the inner structures. In the dye test positive parasites, there were no definite organelles recognizable, as there were in the intact parasites. By the negative staining method, holes (defects) with dark central portions were observed on the surface of the parasites treated with the antibody and the accessory factor, the diameter of the holes measuring about 10–11 nm. These holes, which tended to occur in clusters, were each surrounded by a clear ring.     

On Some Nuisance Parameter Free Uniformly Most Powerful Tests

TAKEUCHI (1969) provides a uniformly most powerful (UMP) one side test for testing the location parameter of the two parameters exponential model when the scale parameter is unknown. The power of his similar size α test depends, however, on the unknown scale parameter. In this case and in more general situations when there exists a sufficient statistic for the nuisance parameter, the theory of generalized THOMPSON's distributions, more specifically, the Thompsonization of a test statistic, LAURENT (1959, 1972) provides a UMP test whose power does not depend on the nuisance parameter. Examples of application of the general nuisance parameter free test procedure include here the truncated exponential, the inverse Gaussian, and the geometric distributions.     

A PCR-based test for the presence of endogenous virus gene evA in chickens

An endogenous virus, denoted ev A, is present at high frequency in all brown egg layer lines. Using inverse polymerase chain reaction (PCR) based on the viral LTR regions, products were obtained containing cellular sequences 5' and 3' to the viral insertion point. PCR of chicken genomic DNA was carried out, using primers chosen from the 5' and 3' cellular sequences and a primer chosen from either the U3 or U5 portions of the viral LTR. Amplification of DNA from birds that did not carry ev A with the primer triplets always gave a single 364bp reaction product, interpreted as representing the flank-to-flank amplification product. Amplification of DNA from known homozygous or heterozygous ev A carriers, with the same primer triplets, always gave both the expected junction product and 364bp product. Therefore, these primer sequences can be used to distinguish ev A carriers from non-carriers but cannot distinguish between homozygous and heterozygous ev A carriers.     

八段锦和五行音乐疗法在缓解新型冠状病毒肺炎患者负性情绪中的疗效分析

摘要 目的：观察八段锦和五行音乐疗法对新型冠状病毒肺炎（COVID-19）患者负性情绪的疗效。方法：收集孝感市第一人民医院就诊的COVID-19患者40例，采用随机数字表法将40例患者随机分为对照组（n=20）和观察组（n=20），观察组接受八段锦和子午流注理论指导下的择时五行音乐疗法进行干预，对照组保持日常生活方式。两组在干预前、干预2周采用焦虑自评量表（SAS）和抑郁自评量表(SDS)分别评定两组实验对象在实验前和干预后的负性情绪的改变，用患者生活质量量表(QLQ-C30)分别对两组患者进行生活质量的测评。结果：接受八段锦和子午流注理论指导下的择时五行音乐疗法的患者负性情绪明显减低，SAS、SDS评分明显较前明显改善，观察组患者SAS评分、SDS评分较对照组降低程度明显，差异存在统计学差异（P＜0.05）；干预后观察组、对照组的角色功能、认知功能、情感功能、躯体功能及生活质量评分均明显改善，接受八段锦和子午流注理论指导下的择时五行音乐疗法的患者在角色功能、认知功能、情感功能、躯体功能及生活质量评分方面均优于对照组，差异存在统计学差异（P＜0.05）。结论：COVID-19患者通过八段锦和五行音乐疗法能够明显减低负性情绪，此种疗法可有效起到"精神内守，病安从来"的安慰作用。     

Unbiased Estimation of Odds Ratio by Using Negative Binomial Plans

The use of the negative binomial distribution in both the numerator and denominator in prospective studies leads to an unbiased estimate of the odds ratio and an exact expression for its variance. Sample sizes that minimize the variance of odds ratio estimates are specified. The variance of the odds ratio estimate is shown to be close to the Cramér-Rao lower bound.