Endocytosis and intracellular fate of liposomes using pyranine as a probe

Lipid vesicles (liposomes) containing pH-sensitive fluorophores were used as probes for the study of liposome entry and intracellular fate. Pyranine [8-hydroxy-1,3,6-pyrenetrisulfonate (HPTS)] was entrapped in the liposome aqueous core during preparation to provide a means of detecting internalization into living cells. HPTS is highly water soluble and shows a strong pH-dependent shift in its fluorescence excitation spectrum. Fluorescence emission (FEM) is slightly pH dependent with excitation (lambda EX) at 350-415 nm but highly pH dependent with lambda EX at 450 nm. Liposomes bearing a net negative charge bound rapidly to CV-1 cells and underwent endocytosis. One hour after liposome addition, high FEM with lambda EX at 413 nm and low FEM with lambda EX at 450 nm suggest that most cell-associated liposomes had been internalized and resided at a mean pH of approximately 6.6. Collapse of cellular H+ gradients with NH4Cl or monensin treatment rapidly and reversibly increased FEM with lambda EX at 450 nm. Direct examination by fluorescence microscopy corroborates the fluorometric data on internalization; over time, FEM remained high with lambda EX at 350-405 nm but decreased with lambda EX at 450-490 nm, showing that all lipid vesicles were internalized within 40 min at 37 degrees C. Acidification of intracellular liposomes increased over 3 h, reaching a minimum value of approximately pH 5.5. HPTS persisted within acidic cellular vesicles for 2-3 days, and cytoplasmic dye was observed infrequently, suggesting that liposome fusion with cellular membranes seldom occurs. Material delivered to the endocytic pathway via lipid vesicles labeled an assortment of intracellular organelles of varying motility and morphology, including dynamic tubular structures whose lumen is acidic.     

The influence of insulin on glucose permeability and metabolism of human granulocytes.

Viable human polymorphonuclear leukocytes isolated from peripheral blood were incubated for 1 h at 37 degrees C with variable concentrations of insulin in a saline medium buffered at pH 7.4. The hormone increased glucose consumption by about 40% without influencing the permeability of the membranes to glucose, whose uptake followed a passive diffusion process. The measurement of intermediates localized activation of glycolysis by insulin, down to 0.36 nM, at the phosphofructokinase step. However, the spectrophotometric measurement showed no activation of phosphofructokinase after preincubation with insulin of either intact granulocytes or crude or ultracentrifuged homogenates. The level of cyclic AMP, which is known to activate phosphofructokinase, was not modified by insulin; cyclic GMP did not activate the enzyme in the granulocyte extracts: neither of the two nucleotides can therefore be considered as a direct messenger of the action of insulin on phosphofructokinase. An important fraction of the extra glucose consumed under the influence of insulin was recovered as neither glycogen nor lactate, nor was it oxidized in the Krebs cycle. It might be assumed to have been converted into glycerolipids. However, insulin produced no detectable accumulation of triglycerides and activated neither the pentose phosphate pathway nor oxidative decarboxylation of pyruvate. The fate of the extra glucose consumed under the influence of insulin therefore remains questionable.     

Malocclusion of the premolar and molar teeth in the guinea pig.

Malocclusions of the premolar and molar teeth of inbred Strain 2/N and Strain 13/N and outbred Dunkin-Harley guinea pigs were examined. A higher incidence of malocclusion observed in the inbred strains suggested a genetic basis for the disorder.     

河口盐沼植物对大型底栖动物群落的影响

通过对长江口崇明东滩盐沼海三棱燕草带大型底栖无脊椎动物进行取样调查,研究了不同盐沼底栖动物群落结构及其多样性特征,分析了盐沼植物群落特征与底栖动物群落的关系。研究发现。（1）沿着河口盐沼海拔梯度,从低位盐沼到高位盐沼,底栖动物群落结构及多样性具有明显的梯度变化;（2）盐沼植被与底栖动物群落有密切的关系,尤其是植株高度,地下部分生物量与底栖动物密度,Shannon-Weiner多样性指数,物种丰度的相关性最显著。（3）盐沼植被影响底栖动物群落,是由于植物的地下部分和地下部分结构导致盐沼生境结构复杂,增加了沉积物表层环境的结构异质性,使生境多样化,给底栖动物提供大量生活空间,（4）不同盐沼带海三陵蔗草群落的差异,提供了盐沼表层地貌的变化,这种变化及植物结构的复杂化,在这一些动物提供拓殖地的同时,也为底栖动物躲避捕食者提供了避护所;（5）盐沼植被可以改变口潮滩生境中的沉积环境,并通过消浪、缓流及调节有机质的输入动态和沉积作用而影响底栖动物群落的组成和结构。     

辽东栎林下和开阔地上短柄五加(Acanthopanax brachpus)无性系种群生长的比较研究

研究了分布于开阔地和辽东栎林下短柄五加无性系种群拆散方式、无性系的形态可塑性和分株的死亡年龄,研究结果表明,开阔地上无性系分株的扩散方式为稀疏线型,根茎长度、无性系分株数目,林下和开阔地差异显著,根茎分枝角度、无性系根茎总长无显著性差异。林下分株的死亡年龄显著小于开阔地上无性系分株的死亡年龄。     

广竹的生物学特性及栽培技术研究

对广竹的生物学特性及栽培技术研究结果表明:广竹的出笋历期较短且非常集中,出笋高峰期集中在4月上旬,出笋数量占全期出笋总数的62.7%;幼竹高生长及枝条生长曲线均呈"S"形,遵循慢—快—慢的节律;此竹种属于混生竹,可移竹繁殖和埋鞭繁殖,前一方法的繁殖效果较好,成活率可达60%,而埋鞭繁殖效果相对较差,成活率只有38%,最佳繁殖时间宜在12月下旬至次年2月进行;根据其各生长阶段的生长规律、无性繁殖栽培技术以及生产实践,提出了主要栽培技术措施。     

呼肠病毒BYD株吸附蛋白σ1的原核表达及其抗原性鉴定

将呼肠病毒BYD株S1片段编码的细胞吸附蛋白基因连接至pET-28a( ),构建表达载体pET-28a( )-S1,转化表达宿主菌E.coli BL21(DE3),分析表达载体能否表达预期大小的重组σ1,并进一步鉴定重组σ1的抗原性。SDS-PAGE实验表明,经IPTG诱导后,表达载体能表达53kD左右蛋白质,与重组σ1的预期大小相符;免疫印迹分析表明重组σ1有较好的抗原性和特异性,可用于呼肠病毒诊断试剂的制备。     

农田小麦在土壤湿润条件下的气孔活动

1984年在北京大屯农业生态系统试验站用MK 3型自动气孔计对土壤湿润、睛朗天气条件下冬小麦气孔传导力的日变化,季节变化、以及叶龄和梯度变化进行了观测。发现叶片传导力(gl)的日变化为双峰曲线,正午gl的降低可能是强光和高的饱和差作用的结果,从而限制了光合潜力的发挥。近轴面传导力(gd)和远轴面传导力(gb)的比(α)不是一个常数,这是因为两面气孔对环境条件的反应不同。gl和冠层传导力(g(?))都以孕穗期为最高。灌浆时期gl和gc有所降低,反映了叶片的过早衰老。这是影响本地区小麦产量的重要因素之一,其原因不仅与土壤水分有关,还与大气条件和病虫害有关。本文还讨论了gl随时龄的变化。观测表明,气孔计可以迅速准确地掌握作物群体的生理动态,有可能用于指导农田灌灌和管理。