Validating Reference Gene Expression Stability in Human Ovarian Follicles,Oocytes, Cumulus Cells,Ovarian Medulla,and Ovarian Cortex Tissue

Human ovarian cells are phenotypically very different and are often only available in limited amounts. Despite the fact that reference gene (RG) expression stability has been validated in oocytes and other ovarian cells from several animal species, the suitability of a single universal RG in the different human ovarian cells and tissues has not been determined. The present study aimed to validate the expression stability of five of the most used RGs in human oocytes, cumulus cells, preantral follicles, ovarian medulla, and ovarian cortex tissue. The selected genes were glyceraldehyde 3-phosphate dehydrogenase (GAPDH), beta-2-microglobulin (B2M), large ribosomal protein P0 (RPLP0), beta-actin (ACTB), and peptidylprolyl isomerase A (PPIA). Overall, the stability of all RGs differed among ovarian cell types and tissues. NormFinder identified ACTB as the best RG for oocytes and cumulus cells, and B2M for medulla tissue and isolated follicles. The combination of two RGs only marginally increased the stability, indicating that using a single validated RG would be sufficient when the available testing material is limited. For the ovarian cortex, depending on culture conditions, GAPDH or ACTB were found to be the most stable genes. Our results highlight the importance of assessing RGs for each cell type or tissue when performing RT-qPCR analysis.     

Effect of Calcium Carbonate Whisker on Impact Toughness of Precast Concrete

The impact toughness and compressive strength of concrete added with calcium carbonate whisker are studied.It is found that calcium carbonate whisker can significantly improve the impact energy consumption at failure of 55 ℃ steam cured concrete,but has limited impact on 90 ℃ steam cured concrete.At the same time,SEM,XRD and LF-NMR were used to study the micro morphology,hydration product components and pore structure of the concrete,and the mechanism of the influence of calcium carbonate whisker on the impact toughness and compressive strength of concrete was analyzed.     

Identification of Host Fruit Volatiles from Flowering Dogwood (Cornus florida) Attractive to Dogwood-Origin Rhagoletis pomonella Flies

Solid-phase microextraction and gas chromatography coupled with electroantennographic detection were used to identify volatiles from fruit of flowering dogwood, Cornus florida, as key attractants for Rhagoletis pomonella flies originating from dogwood fruit. A six-component blend containing ethyl acetate (54.9%), 3-methylbutan-1-ol (27.5%), isoamyl acetate (0.9%), dimethyl trisulfide (1.9%), 1-octen-3-ol (9.1%), and -caryophyllene (5.8%) was identified from flowering dogwood fruit that gave consistent EAD activity. In a flight tunnel assay there was no significant difference in the response of individual dogwood flies exhibiting upwind anemotactic flight to volatile extracts from dogwood fruit and the six-component synthetic mixture. Dogwood flies also displayed significantly greater levels of upwind flight to sources with the dogwood volatile blend than with previously identified volatile blends from domestic apple or hawthorn fruit. Selected subtraction assays showed that the three-component mixture of 3-methylbutan-1-ol, 1-octen-3-ol, and -caryophyllene elicited levels of upwind flight to the source equivalent to the six-component mixture. Our study adds to previous ones showing that populations of Rhagoletis pomonella flies infesting apple, hawthorn, and flowering dogwood fruit are attracted to unique mixtures of fruit volatiles, supporting the hypothesis that host fruit odors could be key traits in sympatric host shifts and establishing host fidelity within members of the Rhagoletis pomonella species complex.     

混合策略改进的乌鸦搜索算法

为了改善乌鸦搜索算法(crow search algorithm,CSA)收敛速度慢、收敛精度不足的问题,提出一种混合策略改进的乌鸦搜索算法(MSCSA).首先在算法运行前期引入tent序列扰动的自适应权重系数,提高算法收敛速度;其次在算法后期引入混合黄金正弦与飞蛾扑火算子,避免算法后期陷入局部最优值;最后通过改进算法的发现概率AP,增加算法的随机性从而提高算法的收敛精度.通过在九个基准函数上对比测试,确定迭代系数的取值,通过Wilcoxon秩和测试验证算法性能.实验结果证明,所提出的MSCSA的性能更为优秀.     

How do mutations at phenylalanine-153 and isoleucine-155 partially suppress the effects of the aspartate-27-->serine mutation in Escherichia coli dihydrofolate reductase?

Several second-site suppressors of the D27S lesion in Escherichia coli dihydrofolate reductase (DHFR) have been identified. The activity of the primary mutant, D27S DHRF, was found to be greatly decreased at pH 7.0, consistent with aspartic acid-27 being critically involved in proton donation during catalysis. Partial suppressors of the D27S mutation have been selected by their ability to confer an increased resistance to trimethoprim upon host E. coli; the suppressors have been identified as F153S or I155N substitutions. D27S+F153S and D27S+I155N DHFRs display 2-3-fold increases in kcat over D27S DHFR values, but only the F153S mutation decreases the Km for dihydrofolate by a factor of 2. Neither double mutant approaches wild-type DHFR activity. Unexpectedly, Phe153 and Ile155 occur on the surface of the protein and are approximately 8 and 14 A distant from the active site. Ile155 is a member of a beta-bulge. A previously identified suppressing mutation, F137S, occurs nearby and is also a member of the same beta-bulge [Howell et al. (1990) Biochemistry 29, 8561-8569]. Clustering of these three second-site mutations indicates this area of the structure may be important in protein function. Conformational changes due to the presence of these suppressing mutations are likely as the F153S and I155N mutations do not affect hydride-transfer rates upon introduction in wild-type DHFR and alterations in circular dichroism spectra are associated with the double-mutant DHFRs.     

Injection-seeded pulsed Ti:sapphire laser with novel stabilization scheme and capability of dual-wavelength operation

A gain-switched, single-frequency titanium-sapphire laser for atmospheric humidity measurements using the differential absorption lidar technique operating in the 820 nm wavelength region is described. The laser is pumped by a frequency-doubled, flashlamp-pumped Nd:YAG laser at a repetition rate of 50 Hz and injection seeded by two external-cavity-diode lasers. The system yields pulses with an energy of 15 mJ and high spectral purity. We describe a novel active injection-locking technique that avoids the problems of established methods like dither-lock or ramp-and-fire. Furthermore, our method opens the possibility to switch between two wavelengths for alternating shots, in contrast to most established techniques that only allow operation at one wavelength.