Determination of motility forces of bovine sperm cells using an "optical funnel"

An optical funnel, a new technique for the evaluation of the force of a microorganism, was applied to the determination of the motility force of bovine sperm cells. In this approach, sperm cells, suspended in an aqueous solution, are introduced into a flow cell, to which radiation pressure is applied from the direction opposite to a medium flow. The sperm cell, which is moving in a stream, is captured by radiation pressure and forced to move to the position at which the force induced by the laser radiation is equal to the force induced by a medium flow. The sperm cell then escapes by its own power on the way to this equilibrium (entrapping) position. The radiation force increases with decreasing distance from the focal point, and as a result, the force of the sperm cell can be determined by measuring the position where the sperm cell escaped against the laser irradiation field. The motility force of the sperm cell was measured in aqueous solution at different pH values and potassium ion concentrations. It was possible to measure more than 250 sperm cells in 3 h. Thus, the optical funnel has potential for use as a rapid and repetitive means for the determination of the motility force of the sperm cell.     

Effects on spermatogenesis in swiss mice of a protein isolated from the roots of Ricinus communis (Linn.) (Euphorbiaceae)

This study was aimed to evaluate the effect on spermatogenesis of a 62 kDa protein (Rp) isolated from 50% ethanolic extract of the root of Ricinus communis in mice. A dose response study in mice revealed that 25mg/kg body weight/day was the most effective dose. Swiss strain mature male mice of 30 days old were divided into two group namely control and Rp treated (25mg/kg body weight/day). The study showed that sperm motility and count were decreased significantly in the treated group as compared to the control. The fertility index of the treated groups was reduced by 100%. The activity of HMG Co A reductase and cholesterol were increased significantly in the treated group. The testicular activities of 3βHSD, 17βHSD, glucose 6-phosphate dehydrogenase and malic enzyme and the level of serum testosterone were decreased significantly in the treated group. The expression of 3βHSD and 17βHSD were decreased and the expression of StAR increased significantly in the treated group as compared to the control. Proteolytic digestion of the native protein with trypsin and chymotrypsin showed that the proteolytic cleavage did not affect the spermicidal action of Rp. Hence this study can be concluded that Rp impaired spermatogenesis in vivo by suppressing the production of testosterone.     

水胺硫磷对玫瑰无须鲃精子的体外毒性作用

通过观察水胺硫磷对玫瑰无须鲃精子运动率和超微结构的影响,发现：(1)室温下玫瑰无须鲃精子激活后30min一级运动率仍维持在78％左右;(2)经5μM、50μM和500μM水胺硫磷处理,精子平均一级运动率随着水胺硫磷浓度的增加和作用时间的延长明显下降;(3)经水胺硫磷处理后,玫瑰无须鲃精子的超微结构发生明显变化,包括质膜光滑程度降低、领和袖套部位的细胞膜局部破裂、线粒体显著膨胀和嵴变模糊或溶解、质膜和核膜溶解以及细胞核电子密度变大。因此,玫瑰无须鲃精子是有机磷农药如水胺硫磷毒性体外快速检测的理想系统。     

Fluid flow and sperm guidance: a simulation study of hydrodynamic sperm rheotaxis

How does a sperm find its way? The study of guidance cues has fascinated sperm biologists and in particular the prospect of rheotaxis, that is a fluid flow orienting the direction of sperm swimming, has been the subject of extensive recent study, as readily motivated by the prospect that such guidance may be active in the mammalian female reproductive tract. For instance, it has been hypothesized that helical sperm flagellar beating is necessary for such guidance, whereas there is an extensive diversity of flagellar beating patterns, with planar sperm beating readily observed in human cells for example. In particular, such cells will not be guided by fluid flow according to hypothesized mechanisms for rheotaxis presented thus far. Here, using simulation methods, we investigate rheotaxis for a wide range of flagellar beat patterns. Providing the virtual sperm firstly does not possess a tightly circling trajectory in the absence of a background flow and secondly, remains within a region of low shear to prevent being washed away by the background flow, rheotaxis is generally observed with the sperm swimming into the flow together with a possible transverse velocity. Tight circling sperm motility, as observed in select hyperactivated sperm and CatSper mutants, is predicted to disrupt the rheotactic response, whereas confinement to low shear regions generally requires boundary accumulation, thus introducing subtleties in the relationship between rheotactic behaviours and the flagellar waveform and sperm characteristics. Nonetheless, such predictions suggest such rheotactic guidance may be more common and robust than previously thought, and we document simple criteria for the presence of rheotaxis that are consistent with our simulations and understanding, as well as reported observations to date.     

Photochemical effect in two-photon laser-induced fluorescence detection of carbon monoxide in hydrocarbon flames

The CO formation as a result of the CO(2) photodissociation at 230.08 nm was observed by using the two-photon laser-induced fluorescence (LIF) method. The measurements were performed in a propane-air combustion product flow and in mixtures of CO(2) and O(2). The temperature dependence of the fluorescence signal caused by CO molecules, produced in the photodissociation of CO(2) molecules under the action of laser radiation at a wavelength of 230.08 nm, was measured at temperatures ranging from 1300 to 2000 K. It is shown that consideration of CO(2) photodissociation under the action of the probing radiation is necessary when one applies the two-photon LIF method for the measurement of small CO concentrations in high-temperature gas mixtures containing CO(2). As an example, a correction is given of the CO concentration profiles measured by the LIF method in the combustion product flow around a cooled metallic plate.     

Detection of paralytic shellfish toxins by a solid-phase inhibition immunoassay using a microsphere-flow cytometry system

Paralytic shellfish poisoning is a toxic syndrome described in humans following the ingestion of seafood contaminated with saxitoxin and/or its derivatives. The presence of these toxins in shellfish is considered an important health threat and their levels in seafood destined to human consumption are regulated in many countries, as well as the levels of other chemically unrelated toxins. We studied the feasibility of immunodetection of saxitoxin and its analogs using a solid-phase microsphere assay coupled to flow cytometry detection in a Luminex 200 system. The technique consists of a competition assay where the toxins in solution compete with bead-bound saxitoxin for binding to an antigonyautoxin 2/3 monoclonal antibody (GT-13A). The assay allowed the detection of saxitoxin both in buffer and mussel extracts in the range of 2.2-19.7 ng/mL (IC(20)-IC(80)). Moreover, the assay cross-reactivity with other toxins of the group is similar to previously published immunoassays, with adequate detection of most analogs except N-1 hydroxy analogs. The recovery rate of the assay for saxitoxin was close to 100%. This microsphere-based immunoassay is suitable to be used as a screening method, detecting saxitoxin from 260 to 2360 μg/kg. This microsphere/flow cytometry system provided similar sensitivities to previously published immunoassays and provides a solid background for the development of easy, flexible multiplexing of toxin detection in one sample.     

Passively driven integrated microfluidic system for separation of motile sperm

This paper describes a self-contained integrated microfluidic system that can separate motile sperm from small samples that are difficult to handle using conventional sperm-sorting techniques. The device isolates motile sperm from nonmotile sperm and other cellular debris, based on the ability of motile sperm to cross streamlines in a laminar fluid stream. The device is small, simple, and disposable yet is an integrated system complete with sample inlets, outlets, sorting channel, and a novel passively driven pumping system that provides a steady flow of liquid; it requires no external power source or controls. The device fulfills a need in clinical settings where small amounts of sperm need to be sorted. It also opens the way for convenient bioassays based on sperm motility including at-home motile sperm tests.     

Evaluation of toxic agent effects on lung cells by fiber evanescent wave spectroscopy

Biochemical changes in living cells are detected using a fiber probe system composed of a single chalcogenide fiber acting as both the sensor and transmission line for infrared optical signals. The signal is collected via evanescent wave absorption along the tapered sensing zone of the fiber. We spectroscopically monitored the effects of the surfactant Triton X-100, which serves as a toxic agent simulant on a transformed human lung carcinoma type II epithelial cell line (A549). We observe spectral changes between 2800-3000 cm(-1) in four absorptions bands, which are assigned to hydrocarbon vibrations of methylene and methyl groups in membrane lipids. Comparison of fiber and transmission spectra shows that the present technique allows one to locally probe the cell plasma membrane in the lipid spectral region. These optical responses are correlated with cellular metabolic activity measurements and LDH (lactate dehydrogenase) release assays that indicate a loss of cellular function and membrane integrity as would be expected in response to the membrane solubilizing Triton. The spectroscopic technique shows a significantly greater detection resolution in time and concentration.     

木瓜蛋白酶动态膜结晶的实验研究

采用聚偏氟乙烯中空纤维微孔膜,考察结晶过程中蛋白质溶液和洗脱液流速的改变以及不同膜面积对溶剂跨膜通量及木瓜蛋白酶晶体尺寸的影响.结果表明:在以pH值为4.7的0.1 mol/L醋酸钠为缓冲溶液、1%的(NH4)3PO4为沉淀剂以及25%的MgCl2溶液作为洗脱液的木瓜蛋白酶动态膜结晶中,当有效膜面积为62.83 cm2,蛋白质溶液和洗脱液流速分别控制在4 744μm/s和1.37×104μm/s的高流速时膜组件的效率得到最大发挥,并得到了较大尺寸的木瓜蛋白酶晶体;当膜面积不同时,溶剂跨膜通量的衰减随着膜面积减小而趋于平缓.     

碳纳米纤维增强聚酰亚胺复合气凝胶的合成与性能EI北大核心CSCD

以4,4′-二氨基二苯醚(ODA)、均苯四甲酸二酐(PMDA)为单体,酸化碳纳米纤维(a-CNF)为增强材料,采用溶胶-凝胶方式成型,运用冷冻干燥技术制备PI复合气凝胶。对复合气凝胶的形貌、隔热、吸波以及压缩性能等进行表征分析。研究结果表明:随着a-CNF含量的增加,PI复合气凝胶的收缩率从45.52%降至35.32%,密度也随之从0.084 g/cm^(3)降至0.069 g/cm^(3),气凝胶孔洞分布呈增大增宽趋势。a-CNF的引入有效抑制了PI复合气凝胶的收缩率,热导率降低;整个体系的导电损耗增加,同时由于气凝胶的多孔结构提供了较好的阻抗匹配,使得PI复合气凝胶的反射损耗(RL)在8.3 GHz达到-9.7 dB。加入质量分数为15%的CNF/PI复合气凝胶压缩强度和压缩模量分别是纯PI气凝胶的近1.5倍和2倍。     

Reconstitution of membrane proteins into polymer-supported membranes for probing diffusion and interactions by single molecule techniques

We have established a robust and versatile analytical platform for probing membrane protein function in a defined lipid environment on solid supports. This approach is based on vesicle capturing onto an ultrathin poly(ethylene glycol) (PEG) polymer brush functionalized with fatty acid moieties and subsequent vesicle fusion into a contiguous membrane. In order to ensure efficient formation of these tethered polymer-supported membranes (PSM), very small unilamellar vesicles (VSUV) containing fluorescent lipids or model transmembrane proteins were generated by detergent depletion with cyclodextrin. Thus, very rapid reconstitution of membrane proteins into PSM was possible in a format compatible with microfluidics. Moreover, surfaces could be regenerated with detergent solution and reused multiple times. Lipid and protein diffusion in these membranes was investigated by fluorescence recovery after photobleaching, single molecule tracking, and fluorescence correlation spectroscopy. Full mobility of lipids and a high degree of protein mobility as well as homogeneous diffusion of both were observed. Quantitative ligand binding studies by solid phase detection techniques confirmed functional integrity of a transmembrane receptor reconstituted into these PSM. Colocomotion of individual ligand-receptor complexes was detected, demonstrating the applicability for single molecule fluorescence techniques.     

Mass spectrometry imaging of small molecules using desorption/ionization on silicon

Development of novel tools to image spatial distribution of small molecules in biological samples is essential in disease diagnosis and biomarker discovery. To simplify sample preparation and reduce background noise in the low-mass region, we describe here the use of a matrix-free mass spectrometric imaging method, i.e., desorption/ionization on silicon (DIOS), for biological surface analysis. The imaging parameters, such as the laser beam diameter and the translation stage movement, were studied and optimized to improve imaging performance. The use of DIOS imaging to map small molecules on mouse liver tissues was demonstrated. In addition, phosphatidylcholine (PC) and propidium iodide (PI) were used as the cell membrane and nucleus markers, respectively, to "visualize" the presence of HEK 293 cells. The reconstructed ion maps of PC and PI were compared with the optical images collected from the same sample using bright-field and fluorescence microscopy. A good correlation of the spatial distribution of cells confirmed the validity of this DIOS imaging approach.     

语音分离技术的研究现状与展望

从计算听觉场景分析和盲源分离两种方法综述了当前语音分离技术的研究现状和发展方向。计算听觉场景分析是用计算机来模拟人类听觉系统的处理机制。它可分为两大类：一类是数据驱动型，特点是信息由低级向高级的单向流动；另一类是图式驱动型，特点是信息由低级向高级和由高级向低级结合的双向流动。最后指出信息双向互流的混合语音分离模式将是未来计算听觉场景分析研究的主要模式：基于听觉和视觉的结合来改善语音分离效果的研究将是未来研究方向之一。此外，盲源分离的欠完全问题．非线性混叠信号的可分离性、非平稳混叠信号的盲分离问题都将需要进一步研究：基于CASA和BSS联合进行语音分离将是未来研究的热点。     

Formulation of a Foaming Vaginal Tablet and Suppository

Abstract

Formulations for a foaming, spermicidal vaginal tablet and a foaming, spermicidal suppository are presented. Some pharmaceutical characteristics peculiar to both dosage forms were measured. A simple method is suggested for the evaluation of the quantity and collapse resistance of the foam.     

Water sorption and temperature effects on the dynamic mechanical behaviour of epoxy-matrix particulates

The effect of water absorption on the dynamic mechanical properties of composite systems consisting of a cold-setting epoxy matrix filled with iron particles was investigated over a wide temperature range. Storage moduli and loss factors of these composites were determined and the effect of water-conditioning temperature was examined. To minimize the complication of higher-order particle-particle interaction, a low filler content was used. Results indicate a strong dependence of dynamic properties on the water-conditioning temperature. It was found that depending on the water-conditioning temperature the presence of water molecules may either enhance or reduce the dynamic properties of the composite. The observed behaviour was similar to the respective behaviour of the same materials when tested in flexure. This behaviour was explained in terms of the dual action of absorbed water as a plasticizer and a crazing agent which deteriorates in a different way the physical and mechanical integrity of the epoxy-matrix composite.     

基于XML构建生物信息二级数据库

提出并实现了一种生物信息二级数据库的构建方法.该方法采用NCBI维护的一级数据库作为核酸和蛋白质序列数据来源,以Oracle 9i作为后台软件,所有的应用程序基于Java开发.通过代理程序自动获取Web数据,并以XML作为中间格式保存,然后通过解析提交到二级数据库中,同时转换成便于Web发布的HTML格式.该方法既方便对语义的机器解析,又有效地保证入库信息的完整性,显著提高了二级数据库的开发效率.     

Structural characterization of individual vesicles using fluorescence microscopy

Extrusion of hydrated lipid suspensions is frequently employed to produce vesicles of uniform size, and the resulting vesicles are often reported to be unilamellar. We describe a method for the quantitative fluorescence image analysis of individual vesicles to obtain information on the size, lamellarity, and structure of vesicles produced by extrusion. In contrast to methods for bulk analysis, fluorescence microscopy provides information about individual vesicles, rather than averaged results, and heterogeneities in vesicle populations can be characterized. Phosphatidylcholine vesicles containing small fractions of biotin-modified phospholipid and fluorescently labeled 7-nitro-2,1,3-benzoxadiazol-4-yl (NBD) phospholipid were immobilized through biotin-avidin-biotin binding to the surface of a biotin-modified glass coverslip. Biotin was attached to the surface in a mixed cyano-terminated silane monolayer. Initial fluorescence intensities for each immobilized vesicle were recorded, and a solution of membrane impermeable quencher was passed through the flow cell to quench the fluorescence of the outer layer. Fluorescence from individual vesicles was measured by fitting the spots to 2-dimensional Gaussian functions. The integrated signals under the peaks yielded a pre- and postquench intensity. From the fractional loss of intensity, the number and structure of the bilayers in individual vesicles could be quantified; the results showed that extruded vesicles exhibit a distribution of size, lamellarity, and structure.     

氮化硼填充聚酰亚胺复合材料的制备及其亚胺化的研究

采用原位聚合法制备聚酰亚胺/氮化硼(PI/BN)复合材料,并对其进行表征分析.通过一系列不同偶联剂、催化剂含量等合成参数的优化,得到最佳性能的薄膜.采用称重法对薄膜的亚胺化及固化程度进行测定,黏度法对聚合物分子量的测定,使用FTIR,DSC和TG对单体、制备过程及最终复合材料的结构和热性能进行表征分析.结果表明,聚酰亚...     

Traceless capping agent for peptide sequencing by partial edman degradation and mass spectrometry

An improved method for the rapid sequence determination of biologically active peptides selected from one-bead-one-peptide combinatorial libraries has been developed. In this method, beads carrying unique peptide sequences were subjected to multiple cycles of partial Edman degradation (PED) by the treatment with a 15-30:1 mixture of phenyl isothiocyanate and N-(9-fluorenylmethoxycarbonyloxy)succinimide (Fmoc-OSU), to generate a series of sequence-specific truncation products (a peptide ladder) for each resin-bound peptide. Following PED, the Fmoc group was removed from the N-terminus and any reacted side chains by piperidine treatment. The sequence of the full-length peptide on each bead was then determined by matrix-assisted laser desorption ionization mass spectrometry. The use of Fmoc-OSU as a traceless capping agent resulted in cleaner MS spectra and improved reliability for sequence assignment. This rapid, sensitive, and inexpensive sequencing method should further expand the utility of combinatorial peptide libraries in biomedical research.