超声波处理提高磁性微小铁葡聚糖颗粒的产量及对纤毛虫细胞的应用

共沉淀法是制备磁性微小铁葡聚糖颗粒的常用方法之一,其简单、易操作,但是温度、pH、氨水滴加速度等多种因素影响粒径的大小,而且很多颗粒聚集成束,因而微小铁葡聚糖颗粒的回收量低。为提高微小铁葡聚糖颗粒的回收量,对反应初产物离心后的沉淀部分进行了超声波处理,使回收量由处理前的0.5g提高到处理后的10g以上。利用回收的微小铁葡聚糖颗粒进行纤毛虫接合对分离,接合率高达95%以上。     

电激诱导DNA摄取的快速评估

英国诺丁汉大学科学家N.W.Blackhall、M.R.Davey及其同事利用流式细胞分类术评估了在荧光素异硫氰酸盐-接合葡聚糖(FITC-葡聚糖)存在下电激水稻原生质体后的瞬时基因表达,并与在携带cat基因的质粒存在下电激水稻原生质体后的瞬时基因表达结果进行了比较。结果发现,FITC-葡聚糖摄取与cat     

硫酸钙作为微小颗粒骨载体的实验研究

目的：研究硫酸钙作为微小颗粒骨载体,解决微小颗粒骨的自身缺点的实际效果,为其临床应用提供依据。方法：将49只日本大耳白兔随机分成4组并通过手术造成双侧桡骨中段1．5cm骨缺损,以植入硫酸钙为载体的自体微小颗粒骨为实验组,同时设立单纯植入自体微小颗粒骨,单纯植入硫酸钙和不植入任何物质的空白对照组。术后4周和8周分别行大体观察。X线摄片,组织学观察,骨生物力学测定。结果：以硫酸钙为栽体的自体微小颗粒骨组比单纯自体微小颗粒骨组及单纯硫酸钙组能更有效地修复骨缺损,单纯颗粒骨组成骨效果优于单纯硫酸钙组。空白组无骨愈合迹象;组织学观察示以硫酸钙为载体的自体微小颗粒骨实验组的成骨效果最好,单纯自体微小颗粒骨组次之;生物力学测定证明以硫酸钙为载体的自体微小颗粒骨实验组的力学强度优于单纯自体微小颗粒骨组及单纯硫酸钙组。结论：硫酸钙是微小颗粒骨的优良载体,以硫酸钙为载体的自体微小颗粒骨成骨速度快,成骨量多,质量高,骨的机械强度高,修复骨缺损的能力较单纯应用微小颗粒骨和硫酸钙强;二者结合可充分发辉各自的优势。     

降钙素葡聚糖颗粒缓释微球的研究

目的：降钙素（一个由32个氨基酸组成的多肽）是治疗骨质疏松的首选药之一。降钙索的劣势是其半衰期过短,需要一天一次注射给药,本实验旨在制备突释小,药物释放浓度稳定的降钙素微球制剂。方法：制备降钙素羧酸葡聚糖颗粒和降钙素硫酸葡聚糖颗粒组合物,分别将其包裹于PLGA微球内,制备成降钙素组合微球,采用C18反相色谱柱研究药物的包封率和体外释放行为。结果：所制得的降钙素葡聚糖颗粒缓释微球体外释放一个月,释放曲线比较完美,接近零级释放。结论：本研究制得的降钙素葡聚糖颗粒缓释组合微球能实现理想的体外缓释效果,为后期药动学实验提供基础。     

德国科学家发明断裂神经重接术

德国科学家最近发明一种可以将断裂的神经纤维重新接合起来并促使其再生长愈合的新型手术方法。不过科学家说 ,该方法尚须在动物身上继续进行试验才可应用于临床。据德新社报道 ,上述研究由德国联邦教研部资助 ,由罗伊特林根和蒂宾根两地的科学家合作完成。参与研究的施洛斯豪尔教授介绍说 ,此前人们普遍认为神经断裂后无法重新接合生长 ,但他们在研究中使用某种特殊的微小“导管”先将断裂神经的两端接合 ,尔后再促进其各自重新生长并最终接合。但科学家没有透露有关“导管”的具体信息以及促进神经组织生长的相关机理等。科学家们表示 ,这…     

小鼠腹腔活化巨噬细胞的酸性磷酸酶测定

在两组Mφ数量相等的条件下,用金氏改良法测定小鼠腹腔Mφ的Acp含量,结果注射葡聚糖后小鼠腹腔Mφ的Acp含量明显高于对照组。同时,从Gomori氏硝酸铅染色的标本上,亦看到用葡聚糖活化后的小鼠腹腔Mφ胞体明显增大,细胞铺展明显,细胞质中的Acp阳性颗粒多而粗大,个别Mφ细胞质中的Acp颗粒呈强阳性反应。结果表明葡聚糖(Dextran T500)是一种较理想的Mφ活化物,具有明显增强Acp活性的作用。     

卵子皮层在受精中的作用

关于卵子皮层的概念,人们有两种不同的意见:一种意见指卵子质膜下的一簿层细胞质,厚度不到5μm,主要成分为皮层颗粒、色素颗粒以及肌动蛋白等,它在结构和功能上与其下的细胞质不同;另一种意见认为还应将质膜-细胞外被复合体包括在内。本文采纳后一种意见。 (一) 皮层在受精时的动态变化向海胆卵细胞质中注入一个微小的铁粒,并在已知强度的磁场内停留一定的时间,通过测铁粒子的移动速率可以测出皮层硬度。实验结果表明,铁粒子在细胞质中央区域比在皮层中运动快,说明皮层处在一种凝胶状态,皮层的     

淀粉分支酶3存在于水稻胚乳的淀粉粒之中

对水稻胚乳淀粉颗粒结合的淀粉分支酶进行了研究.酶活性分析表明水稻胚乳中存在着与淀粉颗粒结合的淀粉分支酶.氨基酸测序分析结果表明结合于水稻胚乳淀粉粒的淀粉分支酶是分子量为84 kD的淀粉分支酶3(rice starch branching enzyme 3; RBE3).从开花后5 d到种子成熟,淀粉颗粒结合的RBE3蛋白都保持较为稳定的含量.Northern 分析表明水稻胚乳发育过程中RBE4最先表达而RBE3和RBE1的表达滞后.综合以上研究结果说明RBE3存在于水稻胚乳的淀粉之中是由于RBE3与淀粉葡聚糖链具有较高亲和性而难以和葡聚糖链解离,进而随着淀粉粒的增长而被其包裹.     

掺铁TiO2纳米颗粒与恒定磁场对HL60白血病肿瘤细胞的灭活效果

通过研究不同浓度、不同磁场作用下TiO2、掺铁TiO2纳米颗粒对HL60白血病细胞活性的影响,以及在接受光照和不接受光照条件下的细胞活性,探讨基于TiO2、掺铁TiO2纳米颗粒作为光敏剂的光动力疗法(PDT)灭活白血病肿瘤细胞的可行性.实验结果表明,纳米颗粒对细胞具有一定的抑制/毒性作用,纳米浓度越大,抑制/毒性作用越明显;磁场对细胞的毒性/抑制作用跟掺铁的浓度以及磁感应强度有关,掺铁纳米组在强磁场作用下对细胞抑制/毒性作用明显;此外,添加了纳米颗粒的PDT灭杀效率要比不添加纳米颗粒的PDT灭杀效率高.     

巨噬细胞膜仿生纳米铁颗粒制备及多形性胶质母细胞瘤MRI成像的初步实验研究

摘要 目的：探讨巨噬细胞膜仿生的纳米铁颗粒（Fe3O4 NCs@MM）对多形性胶质母细胞瘤MRI成像的研究。方法：制备巨噬细胞膜仿生的纳米铁颗粒Fe₃O₄ NCs@MM,利用动态光散射（Dynamic Light Scattering,DLS）和透射电子显微镜（Transmission Electron Microscope,TEM）对其水合动力学粒径、表面电势和形态进行表征。采用SDS-聚丙烯酰胺凝胶电泳（sodium dodecyl sulphate-polyacrylamide gel electrophoresis,SDS-PAGE）评价巨噬细胞膜的完整包覆;紫外可见光谱测定巨噬细胞膜仿生的纳米铁颗粒抗蛋白吸附能力。通过MRI成像系统,分析了含不同浓度的Fe元素（0.1-1.6 mM）的Fe₃O₄ NCs@MM在GSH存在或不存在时的T₁弛豫效应。采用细胞增殖-毒性实验（Cell Counting Kit-8,CCK-8）,测定巨噬细胞膜仿生纳米铁颗粒处理肿瘤细胞24 h后的细胞活性。尾静脉注射巨噬细胞膜仿生纳米铁颗粒至原位胶质母细胞瘤模型中,观察成像效果。结果：巨噬细胞膜仿生的纳米铁颗粒Fe₃O₄ NCs@MM的水合动力学粒径和表面电势分别为 286.5±7.6 nm和-20.7±3.5 mV,且在水溶液中分布均匀,具有较好的单分散性。包覆巨噬细胞膜的纳米铁颗粒具备抗蛋白吸附的能力。MRI成像显示,制备的巨噬细胞膜仿生的纳米铁颗粒Fe₃O₄ NCs@MM为GSH响应型MRI对比剂,具有较好的T₁-加权磁共振成像效果,在尾静脉注射巨噬细胞膜的纳米铁颗粒0.5 h后,肿瘤部位的信号可见增强。结论：巨噬细胞膜仿生的纳米铁颗粒Fe₃O₄ NCs@MM可实现多形性胶质母细胞瘤的MRI成像。     

Mating type differentiation in Tetrahymena thermophila: Strong influence of delayed refeeding of conjugating pairs

Mating type differentiation in Tetrahymena thermophila is known to regularly involve stable hereditary alterations at a single chromosomal locus in the somatic (macro)nucleus. This differentiation is directionally affected by the temperature at which new macronuclei develop after fertilization. We now report large and predictable effects of delayed refeeding of conjugating pairs upon mating type differentiation, particularly among mat-2 homozygotes. The mating types whose frequency is affected the most are IV, VI, and VII, a set different from that most affected by temperature. We interpret our observations to reveal the existence of a second system which can participate in mating type differentiation, with different specificity from the system influenced by temperature under conditions of early refeeding of conjugating pairs. These observations enrich the phenomenology surrounding mating type differentiation in T thermophila and provide additional, easily controllable experimental conditions for the manipulation of mating type frequencies.     

Role of Pili in Bacterial Conjugation

We describe techniques for isolating individual pairs of mating Escherichia coli and observing them under the light microscope. Some pairs achieved close cell-to-cell contact, whereas others remained loosely connected by invisible connections which may be F pili. After 30 min of mating, the pairs were separated and allowed to grow into clones. That many exconjugants derived from "loose"-mating pairs produced recombinants suggests that F pili are involved in the transfer of genetic material. The frequency of formation of recombinants from "close"-mating pairs, however, was significantly higher than that from loose-mating pairs, indicating that a close cell-to-cell contact facilitates chromosome transfer. Death rates of exconjugants from close pairs were also higher than those from loose pairs. Hfr x F(-) matings produced higher death rates than F(+) x F(-) matings. Male cells were found capable of transferring genetic markers to two F(-) cells simultaneously. We conclude that F pili play at least three roles in mating: (i) they initiate contacts between mating pairs; (ii) they facilitate the transfer of genetic material; and (iii) they draw mating cells into a close contact which increases the fertility of the union.     

Virus-like particles in yeast: isolation and infectivity.

Virus-like particles containing electron dense cores are seen in thin sections of intact and degenerated cells of a thermosensitive (ts) strain of Candida tropicalis. A particulate fraction not present in wild-type cells has been isolated from the ts cells disrupted by pressure. The particles are 80-120 nm in diameter. Empty particles with a central cavity are observed. The method of infecting mating pairs of Saccharomyces cerevisiae by partially purified particles is described.     

Perturbance Analysis of Nuclear Determination in Tetrahymena

Mating type frequencies were ascertained among the progeny of crosses of strains A × B, Tetrahymena thermophila under a number of different circumstances. The frequencies are different if the parents are severely starved than if they are well-fed at the time of conjugation; severe starvation of the progeny before the first post-zygotic division has an effect similar to that of starving the parents. Mating type frequencies may also be modified by isolating conjugating pairs into cell extracts before the new macronuclei begin to develop; the changes do not appear to be related in a meaningful way to the mating type of the cells used as a source of the cell extracts. A third means of changing the mating type frequencies involves the exposure of conjugating pairs to CaCl₂ solutions. Finally, changed frequency patterns may appear "spontaneously", and reflect either some as yet unsuspected environmental variable, or else an intrinsic metastable state that conditions the probabilities of mating type fixation. With the exception of the starvation effects, the pattern variations seem to fall into two groups. No satisfactory mechanism to account for these results is yet available.     

Inhibition of Bacterial Conjugation by Ribonucleic Acid and Deoxyribonucleic Acid Male-Specific Bacteriophages

Both deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) male-specific phages, with an F-specific host range, inhibited the bacterial mating process of Escherichia coli. DNA phages prevented the formation of mating pairs but had no effect on mating pairs once they were formed. A step in RNA phage infection, prior to RNA penetration, prevented the formation of mating pairs and, in addition, prevented a fraction of existing mating pairs from completing the mating process. These findings are compatible with the hypothesis that donor cells have a single surface structure involved in both conjugation and male-phage adsorption and that this element is the F pilus.     

Behavior of germinal micronuclei under control of the somatic macronucleus during conjugation in Paramecium caudatum.

In conjugating pairs of Paramecium caudatum, the micronuclear events occur synchronously in both members of the pair. To find out whether micronuclear behavior is controlled by the somatic macronucleus or by the germinal micronucleus, and whether or not synchronization of micronuclear behavior is due to intercellular communication between conjugating cells, the behavior of the micronucleus was examined after removal of the macronuclei from either or both cells of a mating pair at various stages of conjugation. When macronuclei were removed from both cells of a pair, micronuclear development was arrested 1 to 1.5 hr after macronuclear removal. When the macronucleus of a micronucleate cell mating with an amicronucleate cell was removed later than 3 to 3.5 hr of conjugation, that is, an early stage of meiotic prophase of the micronucleus, micronuclear events occurred normally in the operated cell. These results suggest that most micronuclear events are under the control of the macronucleus and that the gene products provided by the macronucleus are transferable between mating cells. One such product is required for induction of micronuclear division and is provided just before metaphase of the first meiotic division of the micronucleus. This factor is effective at a lower concentration in the cytoplasm and/or is more transferable between mating cells than the factors required for other stages. This factor, which seems to be present at least until the stage of micronuclear disintegration, is able to induce repeated micronuclear division as long as it remains active. The factor can act on a micronucleus which has not passed through a meiotic prophase. Moreover, the results suggest the existence of a second factor which is provided by the macronucleus after the first meiotic division that inhibits further micronuclear division.     

Reconstitution of mating active membrane vesicles in Paramecium

Mating-active membrane vesicles isolated from cilia of Paramecium caudatum by the urea-EDTA method were dissolved in 9 mM lithium diiodosalicylate (LIS). Membrane vesicles were reconstituted from the LIS-soluble fraction by dialysis. They showed an ability to induce conjugating pairs without prior occurrence of mating agglutination. The same kind of membrane vesicles was obtained when cilia were treated with 4 mM LIS and stored after removing LIS for two weeks.     

Programmed autophagocytosis accompanying conjugation in the ciliate Stylonychia mytilus

Conjugation and postconjugant development in Stylonychia is accompanied by a period of approximately 80 hr during which the cells are unable to ingest food. This period is one of considerable synthetic activity, encompassing important portions of the development of the new macronucleus. Light- and electron-microscopic observations of conjugating pairs and exconjugant cells reveal a process of autophagocytosis that may provide the supplies of energy (and precursors) necessary for postconjugant developmental events. Small autophagosomes (AP) form in conjugants; degenerating mitochondria are prominent among the inclusions observed in these bodies. Soon after separation of pairs, large dense AP appear, apparently by coalescence and condensation of the smaller AP. These “mature” AP give only a slight acid phosphatase reaction. The number of AP slowly declines during postconjugant development; about 60 hr after separation all the AP have been digested. Several observations suggest that this autophagocytosis is part of the developmental program initiated soon after mating begins: (1) The first indications of AP formation occur while conjugating pairs are still able to feed, and thus cannot be attributed to the stress of starvation; (2) formation of large numbers of AP is rather abrupt, whereas their dissolution is very gradual, covering most of the nonfeeding period; and (3) the pattern of AP formation and dissolution is similar in cells whose new macronuclear development has been prevented by brief hydroxyurea treatment.     

Conjugation in the Ciliate Euplotes octocarinatus: Comparison of Ciliary and Cell Body-Associated Glycoconjugates of Non-mating-Competent, Mating-Competent, and Conjugating Cells

Pair formation in the hypotrichous ciliate Euplotes octocarinatus is a poorly understood phenomenon. In order to obtain information about the molecules involved in this process, we compared ciliary and cell body-associated glycoconjugates of non-mating-competent, mating-competent, and conjugating cells. Detection of glycoconjugates was carried out on Western blots by immunostaining of oxidized, digoxigenin-labeled carbohydrate moieties. Using this method, in both of two complementary mating types a 130-kDa glycoprotein was identified, which appeared on cilia during acquisition of mating competence and was reduced during cell pairing. This suggests an active role of this glycoprotein in ciliary adhesion during pair formation, Additionally, in both of the two mating types a cell body-associated 135-kDa glycoprotein was detected, which is present in non-mating-competent cells as well as in mating-competent cells, but is strongly reduced in conjugating cells. In contrast to the ciliary 130-kDa glycoprotein, the cell body-associated 135-kDa glycoprotein is not surface-exposed [8]. We therefore propose that the cell body-associated glycoprotein is either involved in the preparation for cell fusion or meiosis or that it serves as a cytoplasmic pool for the ciliary 130-kDa glycoprotein.