UPTAKE OF [3H-METHYL]CHOLINE BY MICROSOMAL, SYNAPTOSOMAL, MITOCHONDRIAL AND SYNAPTIC VESICLE FRACTIONS OF RAT BRAIN

Abstract— Microsomal, mitochondrial, synaptosomal and synaptic vesicle fractions of rat brain took up [³H-methyl]choline by a similar carrier-mediated transport system. The apparent K_m for the uptake of [³H-methyl]choline in these subcellular fractions was about 5 × 10^?5 M. Choline uptake was also observed in microsomal fractions prepared from liver and skeletal muscle. Virtually identical kinetic properties for [³H-methyl]choline transport were found in the synaptosomal fractions prepared from the whole brain, cerebellum or basal ganglia. Countertransport of [³H-methyl]choline from the synaptosomal fraction was demonstrated against a concentration gradient. HC-3 was a competitive inhibitor of the uptake of [³H-methyl]choline in brain microsomal, synaptosomal and mitochondria] fractions with respective values for K_i of 4.0, 2.1 and 2.3 × 10^?5 M. HC-15 was a competitive inhibitor of the transport of [³H-methyl]choline in the synaptosomal fraction, with a K_i of 1.7 × 10^?4 M. Upon entry into the microsomal fraction, 74 per cent of the radioactivity could be recovered as unaltered choline, 10 per cent as phosphorylcholine, 1.5 per cent as acetylcholine and 2.5 per cent as phospholipid. Choline acetyltransferase (EC 2.3.1.6) was assayed with [¹⁴C]acetylCoA in synaptosomal fractions prepared from basal ganglia and cerebellum, and in the 31,000 g supernatant fraction of a rat brain homogenate. Enzyme activity was 11-fold greater in the synaptosomal fraction from the basal ganglia than in that from the cerebellum. HC-3 did not inhibit choline acetyltransferase and there was no evidence for acetylation of HC-3. Our findings suggest that choline uptake is a ubiquitous property of membranes in the CNS and cannot serve to distinguish cholinergic nerve endings and their synaptic vesicles.     

Junction potentials,electrode standard potentials,and other problems in interpreting electrical properties of membranes

Summary As background to a detailed analysis of the cation permeation mechanism in rabbit gallbladder epithelium, this paper considers several general problems in interpretation. With regard to liquid junction potentials, the common practice of using saturated KCl bridges was insufficiently accurate for the present purposes because the resulting junctions are time-dependent and poorly understood theoretically. Time-independent and well-defined junction potentials were obtained by arranging all junctions to be of the biionic or single-salt dilution types. The magnitudes of these junction potentials were estimated in three different ways, with good agreement. Recording arrangements using either agar bridges or else Ag/AgCl electrodes also yielded good agreement after appropriate corrections for junction potentials and electrode potentials. The effects of nonelectrolytes on electrode standard potentials were measured. Two experiments were devised to determine whether transepithelially measured electrical properties of the gallbladder refer to a single membrane or to two membranes in series: the potential difference change resulting from a mucosal concentration change was measured as a function of the serosal concentration, and intracellular concentrations were altered by increasing bathing solution osmolalities with an impermeant nonelectrolyte. Both types of experiment indicated that transepithelial measurements are dominated by a single membrane. Small corrections were applied to measured potential differences to take account of unstirred-layer effects with permeant salts.     

Nonsolvent water in liposomes

Summary In liposomes of dimyristoyl lecithin at 40°C, a quantity of water equal to about 11.5 moles water per mole lecithin, or about one-third of the enclosed liposome water or one-fifth of the total pellet water, behaves as if it is unavailable for dissolving sucrose. This phenomenon represents permanent exclusion of sucrose, not simply a space that equilibrates slowly due to the low permeability of sucrose. The amount of nonsolvent water increases with temperature, and is similar to the amount of water bound to the phosphorylcholine groups as estimated by other methods. Nonsolvent water arises from a combination of the forces responsible for salting-out of nonelectrolytes from aqueous solutions by ions, and of steric effects adjacent to a surface. Measured liposome: water partition coeffecients must be corrected for the effect of nonsolvent water.     

基于目标颜色基及梯度方向匹配的菌落分割计数算法

【目的】通过菌落测试片提取菌落并计数,在农业、食品业、医疗卫生等领域中是一项常用且重要的工作。目前,菌落自动计数算法大都是以菌落培养皿为主要工作对象,对菌落测试片适用性较差。另外,目前相关技术在常规的粘连物体分割中有着较好的效果,但在菌落分割计数中,由于菌落本身的形态特征,对粘连菌落分割计数的效果尚不够精准。【方法】为解决此类问题,本文提出一种基于目标颜色基及梯度方向匹配的菌落分割计数算法。首先利用图像中菌落的颜色特征作为基,将图像转换到基空间内,以增强菌落与背景之间的差异,其次利用菌落图像的梯度幅值特征对梯度方向进行滤波,然后通过梯度方向进行匹配,进而将粘连的菌落分割,最后利用非极大值抑制的方法筛选出菌落并计数。【结果】经试验,本研究算法的计数精度可达98.00%,能够满足实际需求。【结论】在针对菌落的目标分割计数中,本研究算法不仅计数精度高,而且具有较好的鲁棒性,在对不同厂家的菌落总数测试片菌落分割计数中均有优异效果;然而在对大面积目标的检测分割中算法的准确率会有所下降,因此,该算法更适合于菌落等小目标的检测分割。     

铜绿假单胞菌fur基因缺失突变株的构建及其表型分析

铁摄取调节蛋白(ferric uptake regulator, Fur)是控制铜绿假单胞菌铁代谢和毒力的关键调节因子。许多课题组尝试构建铜绿假单胞菌fur的缺失突变株均失败,因此铜绿假单胞菌的fur一直被认为是必需基因,这导致其生物学功能一直未得到全面的解析。【目的】构建铜绿假单胞菌fur的缺失突变株,并对该突变株的表型进行分析。【方法】以铜绿假单胞菌PAO1为亲本菌株,通过同源重组的方法构建fur缺失突变株,研究该基因对铜绿假单胞菌生长、铁载体生物合成、抗氧胁迫能力、鞭毛形成、生物被膜形成和毒力等的影响。同时,通过遗传分析对fur缺失突变株生长缺陷表型的原因进行探究。【结果】本研究成功构建了铜绿假单胞菌fur基因的缺失突变株,发现缺失突变fur极大地限制了铜绿假单胞菌的生长能力,并降低了该菌对限铁环境的生长适应性,但不影响该菌对高铁环境的生长适应性。铜绿假单胞菌Δfur的这种生长缺陷表型是细胞生长增殖变慢造成的,而不是诱导细胞死亡引起的。然而,其他异源的fur基因能完全互补Δfur的这种生长缺陷表型,暗示铜绿假单胞菌的Fur蛋白在功能上不存在独特性。尽管Fur与毒素-抗毒素系统PacTA存在功能关联性,但是铜绿假单胞菌Δfur的这种生长缺陷表型却与PacT毒素无关。除了影响铜绿假单胞菌的生长表型,缺失突变fur还使铜绿假单胞菌丧失了对铁载体生物合成的抑制作用,导致该菌对H₂O₂更敏感并丧失了鞭毛的形成能力,同时降低了该菌对大蜡螟幼虫的毒力。此外,缺失突变fur还显著提升了铜绿假单胞菌的胞内环二鸟苷酸(cyclic diguanylate, c-di-GMP)水平,从而诱导pelF和pslA基因的表达,进而促进铜绿假单胞菌生物被膜的形成。【结论】fur是可以缺失的非必需基因,在铜绿假单胞菌的正常生长、铁载体生物合成、抗氧胁迫能力、鞭毛形成、生物被膜形成和毒力等方面都发挥着十分重要的作用,这为针对铜绿假单胞菌的疫苗和抗菌药物开发奠定了基础。     

取食和交配对苹小吉丁飞行能力的影响

【目的】苹小吉丁Agrilus mali是一种严重危害苹果树的钻蛀性害虫。本研究旨在明确苹小吉丁的飞行扩散能力及对其飞行能力产生影响的关键因子。【方法】以SUN-FL型智能飞行磨系统对苹小吉丁不同日龄雌雄成虫的飞行能力进行了测定,同时评价了取食和交配情况对其飞行能力的影响。【结果】苹小吉丁飞行能力均随日龄的增加先增强后逐渐降低,初羽化的成虫飞行能力最低,11日龄成虫的飞行能力最强。雌成虫飞行能力强于雄成虫。在24 h内雌雄成虫的最长飞行距离分别为0.4165和0.3559 km;最长飞行时间分别为0.4582和0.4873 h;最大飞行速度分别为2.4639和1.8561 km/h。取食的3日龄雌成虫的平均飞行距离和平均飞行时间分别为0.047 km和0.048 h,雄成虫的分别为0.044 km和0.042 h;而未取食的雌成虫平均飞行距离和平均飞行时间仅分别为0.016 km和0.013 h,雄成虫的仅分别为0.013 km和0.012 h。交配对飞行能力的影响存在性别差异,已交配雌成虫的飞行能力要强于未交配雌成虫的,而已交配雄成虫的飞行能力却低于未交配雄成虫的。【结论】日龄对苹小吉丁成虫的飞行能力影响作用显著。取食显著提高苹小吉丁雌雄成虫的飞行能力,交配显著提高雌成虫飞行能力。     

论害虫大量发生及其预测(一)

一.对害虫大量发生的一般认识 害虫大量发生(或又称害虫猖獗)系由於发生地的环境条件与害虫生理和形态的统一,而表现於数量上迅速增加的结果,亦即代表某种害虫生存历史中这一时期的最高发展。由於组成环境的各个条件变化速度不同,以及害虫生物性的差异,故猖獗期的间距和每次猖獗期的延续,各有其长短或久暂。研究害虫猖獗过程和造成猖獗原     

东亚飞蝗(Locusta migratoria manilensis Meyen)在中国的发生动态

东亚飞蝗系相当典型的草原型昆虫,它具有草原型昆虫的生活特征,如对温湿度有较大的适应范围,喜光,运动器官发达,能作远距离迁移,有群集习性等。主要分布于中国东部农业区边缘的草原地带,故易于造成农业大灾害,为中国历代著名大害虫之一。