Biophysical correlates of lysophosphatidylcholine- and ethanol-mediated shape transformation and hemolysis of human erythrocytes. Membrane viscoelasticity and NMR measurement

The effects of monopalmitoylphosphatidylcholine (MPPC or lysophosphatidylcholine) and a series of short-chain primary alcohols (ethanol, 1-butanol and 1-hexanol) on cell shape, hemolysis, viscoelastic properties and membrane lipid packing of human red blood cells (RBCs) were studied. For MPPC, the effective membrane concentration to induce the formation of stage 3 echinocytes (8 x 10(6) molecules per cell) was one order of magnitude lower than that needed to induce 50% hemolysis (7 x 10(7) molecules per cell). In contrast, short-chain alcohols induced both shape changes and hemolysis within close concentration range (2.5 x 10(8) to 3.5 x 10(8) molecules per cell). Viscoelastic properties of the RBCs were studied by micropipette aspiration and correlated with shape change. Ethanol-treated RBCs showed a decrease in membrane elastic modulus and an increase in membrane viscosity in the recovery phase at the early stage of shape change. MPPC-treated cells showed the same type of viscoelastic changes, but these were not observed until the formation of stage 2 echinocytes. High-resolution solid-state 13C nuclear magnetic resonance technique was applied to study membrane lipid packing in the ghost membrane by following the chemical shift of hydrocarbon chains. Both MPPC and ethanol caused the 13C-NMR chemical shift to move upfield, indicating that membrane lipids were expanded due to the intercalation of these exogenous molecules. Using data obtained from model compounds, we convert values of chemical shift into a lipid packing parameter, i.e., number of gauche bonds for fatty acyl hydrocarbon chains. Approximately 10(8) interacting molecules per cell are required to induce a detectable change of lipid packing by both MPPC and ethanol. The results indicate that homolysis occurs at a smaller surface area for MPPC- than ethanol-treated RBCs. Our findings suggest that progressive changes in the molecular packing in the membrane lead eventually to hemolysis, but the mode responsible for shape transformation varies with these amphipaths.     

Measurement of 2-deoxyglucose and 2-deoxyglucose 6-phosphate in tissues

The enzymatic methods previously described for 2-deoxyglucose (DG) and 2-deoxyglucose 6-phosphate have been refined and adapted to measurements of brain samples ranging from 50 mg wet weight to less than a microgram dry weight. Procedures for preparing such samples for assay are described. Analytical properties of the enzymes employed are given together with means for overcoming their possible short comings. Emphasis is placed on information useful for employing DG to assess rapid changes in glucose metabolism.     

An improved enzymatic cycle for nicotinamide-adenine dinucleotide phosphate.

We describe the use of column chromatography on the nonpolar adsorbent. Amberlite XAD-2, and on silanized silica gel in the desalting and partial purification of cobalamins. These techniques are both simpler and more versatile than phenol extraction, without sacrificing efficiency. In addition, a solvent system for thin-layer chromatography on silanized silica gel is described which rapidly separates naturally occurring cobalamins.     

Two polypeptide chain constituents of the major protein of the cornified layer of newborn rat epidermis.

The insoluble component of stratum corneum of rat epidermis yields two major bands after extraction with 8 M urea-mercaptoethanol-dithiothreitol. The ratio of these two bands is about 1:1 in terms of protein stain intensity and S-[14C]carboxymethyl label. Both polypeptides were purified to homogeneity by DE-52-cellulose, sodium dodecyl sulfate hydroxylapatite C column chromatography, and preparative DodSO4-polyacrylamide gel electrophoresis. The heavier polypeptide contains 30% alpha helix and the lighter contains 27% alpha helix as determined by circular dichroism studies. Both are sensitive to Pronase and resistant to trypsin, collagenase, and elastase. The lighter chain is stable to pepsin but the heavier can be partially degraded to a smaller polypeptide with a molecular weight similar to that of light chain. Amino acid analysis shows that the light chain contains 12 more tyrosine residues than does the heavy chain, suggesting that the light chain is not generated from the heavy chain. However, the two chains may have a common peptide region. Antiserum prepared against the heavier polypeptide can be completely absorbed by purified lighter polypeptide and vice versa indicating that both chains have some common antigenic determinants. Antibody against either chain can cross-react with the stratum corneum and keratohyalin granules in the epidermis of newborn rat as indicated by fluorescent microscopic observation. Similarly, this antibody also cross-reacts with the cell surface or the contents of spinous and granular cells, and very weakly with basal cells, indicating that the two proteins may be present in the lower strata as well as the stratum corneum.     

Synthesis of myosin heavy and light chains in muscle cultures

The weight ratio of myosin/actin, the myosin heavy chain content as the percentage of total protein (wt/wt), and the kinds of myosin light chains were determined in (a) standard muscle cultures, (b) pure myotube cultures, and (c) fibroblast cultures. Cells for these cultures were obtained from the breast of 11-day chick embryos. Standard cultures contain, in addition to myotubes, large numbers of replicating mononucleated cells. By killing these replicating cells with cytosine arabinoside, pure myotube cultures were obtained. The myosin/actin ratio (wt/wt) for pure myotube, standard muscle, and fibroblast cultures average 3.1, 1.9, and 1.1 respectively. By day 7, myosin in myotube cultures represents a minimum of 7% of the total protein, but about 3% in standard cultures and less than 1.5% in fibroblasts cultures. Myosin from standard cultures contains light chain LC1, LC2, and LC3, with a relative stoichiometry of the molarity of 1.0:1.9:0.5 and mol wt of 25,000, 18,000 and 16,000 daltons, identical to those in adult fast muscle. Myosin from pure myotubes exhibits light chains LC1 and LC2, with a molar ratio of 1.5:1.6. Myosin from fibroblast cultures possesses two light chains with a stoichiometry of 1.8:1.8 and mol wt of 20,000 and 16,000 daltons. Clearly, the faster migrating light chain, LC3, found in standard cultures is synthesized not by the myotubes but ty the mononucleated cells. In myotubes, both the assembly of the sarcomeres and the interaction between thick and thin filaments required for spontaneous contraction occur in the absence of light chain LC3. One set of structural genes for the myosin light and heavy chains appears to be active in mononucleated cells, whereas another set appears to be active in multinucleated myotubes.     

Isolement du vernodalin et du vernolepin a partir de Vernonia guineensis: Authenticité du squelette elemane

The isolation of vernoladin and vernolepin is reported. The authenticity of the occurrence of the elemane skeleton in nature is discussed, and a biosynthetic scheme concerning the genus Vernonia is proposed.     

凋落物和根系对沂蒙山区三种森林恢复方式土壤酸性磷酸酶动力学特征的影响

酸性磷酸酶动力学特征可反映不同底物供应下土壤磷转化状况。以人工恢复的引进种黑松人工林和本地种栓皮栎人工林以及自然恢复的天然次生林为研究对象,开展凋落物添加去除和根系去除对土壤酸性磷酸酶动力学特征的影响研究。结果表明:(1)三种森林恢复方式下土壤酸性磷酸酶活性和最大反应速度(V_max)均为双倍凋落物＞对照＞去除凋落物＞去除根系＞无输入;(2)改变凋落物和根系输入对酸性磷酸酶的半饱和常数(K_m)和催化效率(V_max/K_m)影响不大;(3)酸性磷酸酶活性受速效磷含量的反馈调节;土壤氮可利用性和含水量显著影响酸性磷酸酶活性。改变凋落物和根系输入对引进种黑松人工林土壤有机磷转化能力影响最大,天然次生林次之,本地种栓皮栎人工林最稳定。根系对土壤酸性磷酸酶动力学特征的影响大于凋落物。其结果可为暖温带森林恢复,应对气候变化和森林防火、收集凋落物等管理措施提供理论依据。     

菜粉蝶幼虫、蛹和成虫的雌雄形态鉴别

利用体视显微镜观察、测量、记录菜粉蝶不同虫态雌雄个体间的形态特征差异。雌雄菜粉蝶幼虫、蛹和成虫均存在显著的雌雄二型特征,3龄之后雄性幼虫的第6腹节有一对肾型黑斑,雌性幼虫无此斑。雄蛹第9腹节有生殖孔和一条短纵裂缝,裂缝两侧有凹凸不平的半圆形瘤状突起;雌蛹腹部第8、9腹节分别有生殖孔和产卵孔,且两孔之间亦有一条纵裂缝。雌蝶前翅背面有一对上下排列的黑圆斑,其腹部末端外生殖器为圆筒型;雄蝶前翅背面一对黑斑相对较小,颜色较浅,尤其下斑为浅灰色,腹部末端具钳状外生殖器。本研究揭示了菜粉蝶幼虫、蛹及成虫不同虫态的雌雄二型差异,比较研究并提出了便于快速鉴别雌雄个体的典型特征和识别方法。     

强化产电微生物与电极间电子传递速率的研究进展

产电微生物是微生物燃料电池、电解池和电合成等微生物电化学技术(Microbial electrochemical technologies,METs)的研究基础.产电微生物与电极界面间的胞外电子传递(Extracellular electron transfer,EET)效率低以及生物被膜形成能力弱限制了METs在有机...     

四合木种群的生态适应性

通过四合木异地播种生长量比较、以及四合木对土壤的适应性和对水热综合因子的适应性分析,得到如下结果：（1）四合木生长量高低的顺序为乌海区的东胜土（12．12g）〉乌海区的乌海土（10．62g）〉东胜区的乌海土（3．10g）〉东胜区的东胜土（2．59g）。同一地区不同土壤间四合木生长量之间没有差异（P≥0．05）,不同地区相同土壤间的生长量之间却有十分显著的差异,说明了土壤条件对四合木的生存影响是次要的,气候条件是保证四合木生存的最重要的条件。（2）四合木分布区与异地保护区之间9种土壤微量元素具有差异,但由于四合木植株中的微量元素含量均高于土壤中的有效量,说明四合木对微量元素具有富集作用,土壤微量元素含量不是四合木成活与生长的限制因子。（3）四合木分布区与异地保护区之间各月平均温度差异显著;全年气温日较差以及温暖指数、寒冷指数、湿润指数和水热综合因子指数差异显著;生长季地表地温昼夜温差具有特殊性,这可能是异地保护不易成功的限制因子之一。