Multifunctional picoliter droplet manipulation platform and its application in single cell analysis

We developed an automated and multifunctional microfluidic platform based on DropLab to perform flexible generation and complex manipulations of picoliter-scale droplets. Multiple manipulations including precise droplet generation, sequential reagent merging, and multistep solid-phase extraction for picoliter-scale droplets could be achieved in the present platform. The system precision in generating picoliter-scale droplets was significantly improved by minimizing the thermo-induced fluctuation of flow rate. A novel droplet fusion technique based on the difference of droplet interfacial tensions was developed without the need of special microchannel networks or external devices. It enabled sequential addition of reagents to droplets on demand for multistep reactions. We also developed an effective picoliter-scale droplet splitting technique with magnetic actuation. The difficulty in phase separation of magnetic beads from picoliter-scale droplets due to the high interfacial tension was overcome using ferromagnetic particles to carry the magnetic beads to pass through the phase interface. With this technique, multistep solid-phase extraction was achieved among picoliter-scale droplets. The present platform had the ability to perform complex multistep manipulations to picoliter-scale droplets, which is particularly required for single cell analysis. Its utility and potentials in single cell analysis were preliminarily demonstrated in achieving high-efficiency single-cell encapsulation, enzyme activity assay at the single cell level, and especially, single cell DNA purification based on solid-phase extraction.     

The label-free unambiguous detection and symbolic display of single nucleotide polymorphisms on DNA origami

Single nucleotide polymorphisms (SNPs) are the most common genetic variation in the human genome. Kinetic methods based on branch migration have proved successful for detecting SNPs because a mispair inhibits the progress of branch migration in the direction of the mispair. We have combined the effectiveness of kinetic methods with atomic force microscopy of DNA origami patterns to produce a direct visual readout of the target nucleotide contained in the probe sequence. The origami contains graphical representations of the four nucleotide alphabetic characters, A, T, G and C, and the symbol containing the test nucleotide identity vanishes in the presence of the probe. The system also works with pairs of probes, corresponding to heterozygous diploid genomes.     

保加利亚铀矿勘探史例和新的铀矿勘探地区

保加利亚于1945年开始找铀,并于50年代后期发现了第一批铀矿床。1946年成立了保加利亚-苏联联合开发公司。经过45年的勘探,对整个保加利亚领土(11万km~3)均进行了盖格米勒计数器航空伽玛测量。对80％的领土进行了地面伽玛测量,总共打了1800多万米钻孔和640km的平巷工程。在45年期间由于进行了上述工作,结果共发现了35个矿床。其中某些矿床,如Buhovo、Eleshnitza和Momino可以说是矿区,由几个矿床组成。这些矿区是由中等规模矿床组成,已计算的储量为5000—20000tU。其余的32个矿床为小矿床,储量不超过5000tU。有7个矿床处于勘探阶段,14个正在开采,14个已采完。位于保加利亚西南部的2个水冶厂处理来自Buhovo脉型矿床的矿石和Eleshnitza砂岩型矿床的矿石。2个水冶厂的生产能力足以处理全部开采的矿石以及来自原地浸出的载铀树脂。在保加利亚原地浸出技术应用很广泛,7(?)的铀是用这一方法回收的。75％的储量赋存于砂岩型矿床中。从Buhovo、Partisanska Poliana、Beli Iskar、Smolian等脉型矿床中开采的是高品位矿石。保加利亚的铀储量、矿山和水冶厂的生产能力足以满足6个核电站(总装机容量3.54GW)对铀燃料的需要。     

钢轨导电接头

导电接头系普通钢轨接头,只在钢轨接头缝隙处用插塞方式或焊接方式连接以改善导电性。插塞式连接用两根φ5mm的金属丝组成,其末端接在金属插头内,插头塞在轨腰的孔眼中,孔眼是用φ9.8m的电钻钻成的,距轨端470mm。为导信号电流,应用插塞式连接,也可用φ6mm、长200mm的钢丝焊接到钢轨上。     

Estimation of the Ballistic Effectiveness of 3,4-and 3,5-Dinitro-1-(trinitromethyl)-1H-Pyrazoles as Oxidizers for Composite Solid Propellants

The experimental values of the enthalpy of formation of two isomeric 3,4-and 3,5-dinitro-1-(trinitromethyl)-1H-pyrazoles have been obtained(261.5±5.0and 246.4±6.7kJ/mol for crystalline 3,4-and 3,5-dinitro-1-(trinitromethyl)-1H-pyrazoles,respectively).The ballistic effectiveness of these potential oxidizers in composite solid propellants was studied.It is shown that these two oxidizers may be successfully applied in metal-free compositions or with a small content of metal.For the bottom stage 3,4-dinitro-1-(trinitromethyl)-1H-pyrazole is a bit better than 3,5-dinitro-1-(trinitromethyl)-1H-pyrazole,for the upper stage the both oxidizers show the equal ballistic parameters.These oxidizers allow to create metal-free solid composite propellants with the binder percentage not lower than 19%(volume fraction),with I3spequal to 256.5-257.0sat density equal to 1.72-1.74g/cm~3.     

Nutritional quality and acceptability of a product formulated with mechanically deboned poultry meat,plasma and bovine red cell

The purpose of this research was to evaluate the nutritional quality and acceptability of a meat product formulated with mechanically deboned poultry meat (40%), bovine plasma (40%) and bovine red cells (3%) as major ingredients. Proteins, fat, humidity and ash were determined by AOAC procedure, iron and calcium by Atomic Absorption Spectrometry and essential amino acids by HPLC. Biological evaluation was performed using digestibility and PER procedures. To evaluate the acceptability (flavor, color and odor) of the product were used 277 children. Results indicated that 100 g of the formulated meat product contain 10.7 g of proteins, 4.4 g of fat, 68.6 g of humidity, 2.9 g of ash, 4.9 mg of iron, 46.6 mg of calcium. Essential amino acids are above the FAO requirements for a highly nutritional protein digestibility (92.4%) and PER (2.18). Acceptability was 91.3% for flavor, 81.2% for color and 92.4% for odor. The high nutritional quality and acceptability along with the low cost make this meat product suitable for government social programs.     

Short-term effects of selenium supplementation of cows' feed on the content and distribution of selenium, copper and zinc in bovine milk, whey and blood plasma

The effect of selenium supplementation of feed on the Se content in bovine milk, whey and plasma, and on the distribution of Se, Zn and Cu in whey and plasma was investigated. In a cross-over study two groups of cows were given a basal feed with 0.16 ppm selenite (approx. 3 mg Se/d) with or without 25 mg yeast Se/d for 2 weeks. In the supplemented group the Se content increased 10-fold in milk, 10-fold in whey and 2-fold in plasma, and after the cessation of the supplementation, selenium in milk decreased with a calculated half-life of 3.5 d. In another experiment, two groups of cows were given either 100 mg yeast Se/d for 1 week or only the basal feed. The increase in Se content in both whole and defatted milk was 40-50-fold, and in whey it was approx. 20-fold. Size-exclusion chromatography of whey using inductively coupled plasma mass spectrometry for detection showed that supplementation increased the proportion of Se in the beta-lactoglobulin-alpha-lactalbumin fraction. Distribution of Cu and Zn was essentially unaffected. In plasma, supplementation increased the Se content in all major Se fractions like selenoprotein P, albumin and low-molecular-weight compounds, but the distribution profiles of Zn and Cu underwent no major changes. The study showed for the first time the rapid kinetics of the Se increase and decrease in milk after the initiation and cessation of supplementation, respectively, and the preferential appearance of Se in the beta-lactoglobulin-alpha-lactalbumin fraction of whey. Milk highly enriched in selenium will be a useful tool for different research purposes.     

检测和估值引论

本文对检测问题和估值问题作了一般性介绍。在第一部分里简要地说明了两种不同的观点。随后说明在高斯噪声情况下,检测和估值之间有密切的联系,并给出了一些实例。本文特别对自适应检测提出了一些想法,它需要纯参考噪声的估值。文中对纯参考噪声给出了定义,并用它设计了一种检测接收机。     

Oxidative Stability of Polyunsaturated Edible Oils Mixed With Microcrystalline Cellulose

The oxidative stability of mixtures of edible oils containing polyunsaturated fatty acids (PUFA) and microcrystalline cellulose (MCC) was investigated. The mixtures studied consisted of oils of either camelina (CAM), cod liver (CLO), or salmon (SO) mixed with either colloidal or powdered MCC. A 50:50 (w/w) ratio of oil:MCC resulted in an applicable mixture containing high levels of PUFA edible oil and dietary fiber. The oxidative stability of the formulated mixtures and the pure oils was investigated over a period of 28 days. The peroxide value (PV) was assessed as a parameter for primary oxidation products and dynamic headspace gas chromatography mass spectrometry (GC/MS) was used to analyze secondary volatile organic compounds (VOC). CAM and the respective mixtures were oxidatively stable at both 4 and 22 °C during the storage period. The marine oils and the respective mixtures were stable at 4 °C. At 22 °C, an increase in hydroperoxides was found, but no increase in VOC was detected during the time-frame investigated. At 42 °C, prominent increases in PV and VOC were found for all oils and mixtures. Hexanal, a common marker for the degradation of n-6 fatty acids, propanal and 2,4-heptadienal (E,E), common indicators for the degradation of n-3 fatty acids, were among the volatiles detected in the headspace of oils and mixtures. This study showed that a mixture containing a 50:50 ratio of oil:MCC can be obtained by a low-tech procedure that does not induce oxidation when stored at low temperatures during a period of 1 month.     

沿海城市的规划结构

近100-150年,苏联许多沿海的大城市(如塞瓦斯托波尔、刻赤、诺沃罗西斯克、日丹诺夫、塔干罗格等城市)都是作为商业-工业中心城市发展起来的。因此。它们的规划结构几乎仅仅着眼于安排常住人口和布置一些工业、仓库设施。休疗养区在这里是自发地形成的,其占地面积,目前还不大。原有的公共交通和城市干道系统主要为当地居民服务。