猕猴胚胎干细胞的分离培养和鉴定

目的　从体外培养成熟囊胚中分离并鉴定猕猴胚胎干细胞(embryonic stem cell , ES cell) 。方法　猕猴卵母细 胞经体外成熟培养、体外受精和早期胚胎体外成熟培养后,获得猕猴囊胚。当囊胚由透明带自然孵出后,用细玻璃针剥离 囊胚中的内细胞团(inner cell mass , ICM) 并与饲养细胞进行共培养。由ICM分离,培养并鉴定胚胎干细胞集落。结果　由 4 只FSH 超排猕猴中共取得92 个处于GV 期的猕猴卵母细胞,选取其中的22 个用HECM210 培养基培养后,获得6 个高质 量的囊胚,由此6 个囊胚中分离得到3 个内细胞团,并由此最终获得1 株猕猴ES 细胞,即RS5 细胞。RS5 细胞具高比例核P 质比,核仁多,其细胞集落边缘平整,其内各单个细胞清晰。经约5 个月的连续传代后,仍保持了正常二倍体的核型,其染 色体数目为42 条。碱性磷酸酶细胞组织化学染色为阳性,说明RS5 细胞为未分化态的胚胎干细胞。经高密度和长时间 培养后,RS5 细胞可进一步分化为多种类型细胞。结论　RS5 细胞株具有自我更新能力和多分化潜能,属于胚胎干细胞。

Kinetics and mechanism of Y(III) extraction with CA-100 using a constant interfacial cell with laminar flow

The Yttrium(III) extraction kinetics and mechanism with secnonylphonoxy acetic acid (CA-100) were investigated by a constant interfacial cell with laminar flow. The studies of interfacial tension and solubility of extractant and effects of the stirring rate, temperature, specific interfacial area and species concentration on the extraction rate showed that the extraction regime was dependent on the extraction conditions and the most probable reaction zone was at the liquid-liquid interface. The rate equation of extracting yttrium by CA-100 in heptane was Rf = k[Y3+]((a))[H(2)A(2)]((o))(0.88)[H+]((a))(-1.08).

Mass transfer kinetics of neodymium(III) extraction by calix[4]arene carboxylic acid using a constant interfacial area cell with laminar flow

BACKGROUND: Thermodynamics and kinetics data are both important to explain the extraction property. In order to develop a novel separation technology superior to current extraction systems, many promising extractants have been developed including calixarene carboxylic acids. The extraction thermodynamics behavior of calix[4]arene carboxylic acids has been reported extensively. In this study, the mass transfer kinetics of neodymium(III) and the interfacial behavior of calix[4]arene carboxylic acid were investigated.

Mass transfer kinetics of yttrium(III) using a constant interfacial cell with laminar flow. Part II. Extraction with Cyanex 272

The yttrium(III) extraction kinetics and mechanism with bis-(2,4,4-trimethyl-pentyl) phosphinic acid (Cyanex 272, HA) dissolved in heptane have been investigated by constant interfacial cell with laminar flow. The data has been analyzed in terms of pseudo-first order constants. Studies on the effects of stirring rate, temperature, acidity in aqueous phase, and extractant concentration on the extraction rate show that the extraction regime is dependent on the extraction conditions. The plot of interfacial area on the rate has shown a linear relationship. This fact together with the strong surface activity of Cyanex 272 at heptane-water interfaces has made the interface the most probable location for the chemical reactions. The forward, reverse rate equations and extraction rate constant for the yttrium extraction with Cyanex 272 have been obtained under the experimental conditions. The rate-determining step has been also predicted from interfacial reaction models. The predictions have been found to be in good agreement with the rate equations obtained from experimental data, confirming the basic assumption that the chemical reaction is located at the liquid-liquid interface.

Mass transfer kinetics of yttriium(III) using a constant interfacial cell with laminar flow. Part I. Extraction with Cyanex 302

The interfacial tension is measured for Cyanex 302 in heptane and adsorption parameters are calculated according to Gibbs equation and Szyskowski isotherm. The results indicate that Cyanex 302 has a high interfacial activity, allowing easy extraction reaction to take place at the liquid-liquid interface. The extraction kinetics of yttrium(III) with Cyanex 302 in heptane are investigated by a constant interfacial cell with laminar flow. The effects of stirring rate, temperature and specific interfacial area on the extraction rate are discussed. The results suggest that the extraction kinetics is a mixed regime with film diffusion and an aqueous one-step chemical reaction proposed to be the rate-controlling step. Assuming the mass transfer process can be formally treated as a pseudo-first-order reversible reaction with respect to the metal cation, the rate equation for the extraction reaction of yttrium(III) with Cyanex 302 at pH <5 is obtained as follows:R-f = 10(-7.85)[Y(OH)(2)(+)]((a))[H(2)A(2)]((o))(1.00)[H+]((a))(-1.00)Diffusion parameters and rate constants are calculated through approximate solutions of the flux equation.

Quantifying Cell Binding Kinetics Mediated By Surface-Bound Blood Type B Antigen To Immobilized Antibodies

Cell adhesion is crucial to many biological processes, such as inflammatory responses, tumor metastasis and thrombosis formation. Recently a commercial surface plasmon resonance (SPR)-based BIAcore biosensor has been extended to determine cell binding mediated by surface-bound biomolecular interactions. How such cell binding is quantitatively governed by kinetic rates and regulating factors, however, has been poorly understood. Here we developed a novel assay to determine the binding kinetics of surface-bound biomolecular interactions using a commercial BIAcore 3000 biosensor. Human red blood cells (RBCs) presenting blood group B antigen and CM5 chip bearing immobilized anti-B monoclonal antibody (mAb) were used to obtain the time courses of response unit, or sensorgrams, when flowing RBCs over the chip surface. A cellular kinetic model was proposed to correlate the sensorgrams with kinetic rates. Impacts of regulating factors, such as cell concentration, flow duration and rate, antibody-presenting level, as well as pH value and osmotic pressure of suspending medium were tested systematically, which imparted the confidence that the approach can be applied to kinetic measurements of cell adhesion mediated by surface-bound biomolecular interactions. These results provided a new insight into quantifying cell binding using a commercial SPR-based BIAcore biosensor.

A Novel Counter Sheet-Flow Sandwich Cell Culture Device For Mammalian Cell Growth In Space

Cell culture and growth in space is crucial to understand the cellular responses under microgravity. The effects of microgravity were coupled with such environment restrictions as medium perfusion, in which the underlying mechanism has been poorly understood. In the present work, a customer-made counter sheet-flow sandwich cell culture device was developed upon a biomechanical concept from fish gill breathing. The sandwich culture unit consists of two side chambers where the medium flow is counter-directional, a central chamber where the cells are cultured, and two porous polycarbonate membranes between side and central chambers. Flow dynamics analysis revealed the symmetrical velocity profile and uniform low shear rate distribution of flowing medium inside the central culture chamber, which promotes sufficient mass transport and nutrient supply for mammalian cell growth. An on-orbit experiment performed on a recovery satellite was used to validate the availability of the device.

Control of lunar and martian dust - Experimental insights from artificial and natural cyanobacterial and algal crusts in the desert of Inner Mongolia, China

Studies on the colonization of environmentally extreme ground surfaces were conducted in a Mars-like desert area of Inner Mongolia, People's Republic of China, with microalgae and cyanobacteria. We collected and mass-cultured cyanobacterial strains from these regions and investigated their ability to form desert crusts artificially. These crusts had the capacity to resist sand wind erosion after just 15 days of growth. Similar to the surface of some Chinese deserts, the surface of Mars is characterized by a layer of fine dust, which will challenge future human exploration activities, particularly in confined spaces that will include greenhouses and habitats. We discuss the use of such crusts for the local control of desert sands in enclosed spaces on Mars. These experiments suggest innovative new directions in the applied use of microbe-mineral interactions to advance the human exploration and settlement of space.

Man-made desert algal crusts as affected by environmental factors in Inner Mongolia, China

Man-made desert algal crusts were constructed on a large scale (3000m(2)) in Inner Mongolia, China. Microcoleus vaginatus was mass cultivated and inoculated directly onto unconsolidated sand dune and irrigated by automatic sprinkling micro-irrigation facilities. The crusts were formed in a short time and could resist the erosion of winds and rainfalls 22 days after inoculation. The maximum biomass in the man-made algal crusts could also reach 35 mu g Chl a/cm(2) of soil. Effects of environmental factors such as temperature, irrigation, rainfall and soil nutrients on algal biomass of man-made algal crusts were also studied. It was found that rainfalls and lower light intensity had significantly positive effects on the biomass of man-made algal crusts. The preliminary results suggested that man-made algal crusts could be formed rapidly, and thus it might be a new feasible alternative method for fixing unconsolidated sand. (c) 2006 Elsevier Ltd. All rights reserved.

EROD activities in a primary cell culture of grass carp (Ctenopharyngodon idellus) hepatocytes exposed to polychlorinated aromatic hydrocarbonas

Aryl hydrocarbon (Ah) receptor (Ah-agonist) effects of environmental samples containing polychlorinated aromatic hydrocarbons were evaluated using a 7-ethoxyresorufin-O-deethylase (FROD) assay of a primary hepatocyte culture from grass carp (Ctenopharyngodon idellus). The results were compared with those obtained from the assay using the rat hepatoma cell line H4IIE and chemical analysis using high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS). A dose-response relationship was observed between the EROD activities, either from primary hepatocyte culture assay or from H4IIE assay, and concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The results showed that the assay based on the H4IIE cell line (EC50 = 0.83 mug/mL) is more sensitive to TCDD than the assay based on primary hepatocyte Culture (EC50 = 9.7 pg/mL). In tests of environmental samples, the results from the assay using primary hepatocyte culture were comparable to those from the assay using the H4IIE cell line and chemical analysis of concentrations of mixtures of polychlorinated dibenzo-p-dioxin and dibenzofuran (PCDD/PCDF). The lack of a change in the activities of glutathione-S-transferase (GST) and lactate dehydrogenase (LDH) in cell culture upon exposure to TCDD indirectly indicates that the compound is persistent to biodegradation in the cell culture system. (C) 2004 Elsevier Inc. All rights reserved.

35-PERCENT EFFICIENT NONCONCENTRATING NOVEL SILICON SOLAR-CELL

An energy conversion efficiency of 35% was obtained at 1-sun, air mass 1.5 for a novel silicon cell having an area of 2.3 X 2.3 mm2 . cell. The critical feature of the cell structure is the inclusion of local defect layers near a p-n junction. The local defect layers were proven to hold the key to achieving the exceptionally high efficiency of the novel cell fabricated via noncomplex processing.

Immobilized metal-ion chelating capillary microreactor for peptide mapping analysis of proteins by matrix assisted laser desorption/ionization-time of flight-mass spectrometry

Peptide mass mapping analysis, utilizing a regenerable enzyme microreactor with metal-ion chelated adsorption of enzyme, combined with matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) was developed. Different procedures from the conventional approaches were adopted to immobilize the chelator onto the silica supports, that is, the metal chelating agent of iminodiacetic acid (IDA) was reacted with glycidoxypropyltrimethoxysilane (GLYMO) before its immobilization onto the inner wall of the fused-silica capillary pretreated with NH4HF2. The metal ion of copper and subsequently enzyme was specifically adsorbed onto the surface to form the immobilized enzyme capillary microreactor, which was combined with MALDI-TOF-MS to apply for the mass mapping analysis of nL amounts of protein samples. The results revealed that the peptide mapping could routinely be generated from 0.5 pmol protein sample in 15 min at 50degreesC, even 20 fmol cytochrome c could be well digested and detected.

Modelling raindrop impact and splash erosion processes within a spatial cell: a stochastic approach

A new approach is proposed to simulate splash erosion on local soil surfaces. Without the effect of wind and other raindrops, the impact of free-falling raindrops was considered as an independent event from the stochastic viewpoint. The erosivity of a single raindrop depending on its kinetic energy was computed by an empirical relationship in which the kinetic energy was expressed as a power function of the equivalent diameter of the raindrop. An empirical linear function combining the kinetic energy and soil shear strength was used to estimate the impacted amount of soil particles by a single raindrop. Considering an ideal local soil surface with size of I m x I m, the expected number of received free-failing raindrops with different diameters per unit time was described by the combination of the raindrop size distribution function and the terminal velocity of raindrops. The total splash amount was seen as the sum of the impact amount by all raindrops in the rainfall event. The total splash amount per unit time was subdivided into three different components, including net splash amount, single impact amount and re-detachment amount. The re-detachment amount was obtained by a spatial geometric probability derived using the Poisson function in which overlapped impacted areas were considered. The net splash amount was defined as the mass of soil particles collected outside the splash dish. It was estimated by another spatial geometric probability in which the average splashed distance related to the median grain size of soil and effects of other impacted soil particles and other free-falling raindrops were considered. Splash experiments in artificial rainfall were carried out to validate the availability and accuracy of the model. Our simulated results suggested that the net splash amount and re-detachment amount were small parts of the total splash amount. Their proportions were 0.15% and 2.6%, respectively. The comparison of simulated data with measured data showed that this model could be applied to simulate the soil-splash process successfully and needed information of the rainfall intensity and original soil properties including initial bulk intensity, water content, median grain size and some empirical constants related to the soil surface shear strength, the raindrop size distribution function and the average splashed distance. Copyright (c) 2007 John Wiley & Sons, Ltd.

A new heteroleptic ruthenium sensitizer enhances the absorptivity of mesoporous titania film for a high efficiency dye-sensitized solar cell

A heteroleptic polypyridyl ruthenium complex, cis-Ru(4,4'-bis(5-octylthieno[3,2-b]thiophen-2-yl)-2,2'-bipyridine)(4,4'-dicarboxyl-2,2'-bipyridine)(NCS) 2, with a high molar extinction coefficient of 20.5 x 10(3) M-1 cm(-1) at 553 nm has been synthesized and demonstrated as a highly efficient sensitizer for a dye-sensitized solar cell, giving a power conversion efficiency of 10.53% measured under an irradiation of air mass 1.5 global ( AM 1.5G) full sunlight.

Extraction kinetics of thorium(IV) with primary amine N1923 in sulfate media using a constant interfacial cell with laminar flow

Extraction kinetics of thorium(IV) with primary amine N1923 in sulfate media has been investigated by a constant interfacial cell with laminar flow. Studies of interfacial tension and effects of the stirring rate, temperature, and specific interfacial area on mass transfer rate show that the most probable reaction zone takes place at the liquid-liquid interface. According to the experimental data correlated as a function of the concentration of the relevant species involved in the extraction reaction, the rate equation of extracting thorium has been obtained as follows: -d[Th(IV)]((o))/dt = 10(-3.10)center dot[Th(IV)](0.89)center dot[(RNH3)(2)SO4](0.74).