Vitrification of one-cell mouse embryos in cryotubes

Preventing intracellular ice formation is essential to cryopreserve cells. Prevention can be achieved by converting cell water into a non-crystalline glass, that is, to vitrify. The prevailing belief is that to achieve vitrification, cells must be suspended in a solution containing a high concentration of glass-inducing solutes and cooled rapidly. In this study, we vitrified 1-cell mouse embryos and examined the effect of the cooling rate, the warming rate, and the concentration of cryoprotectant on cell survival. Embryos were vitrified in cryotubes. The vitrification solutions used were EFS20, EFS30, and EFS40, which contained ethylene glycol (20, 30 and 40% v/v, respectively), Ficoll (24%, 21%, and 18% w/v, respectively) and sucrose (0.4 0.35, and 0.3 M, respectively). A 5-μl EFS solution suspended with 1-cell embryos was placed in a cryotube. After 2 min in an EFS solution at 23 °C, embryos were vitrified by direct immersion into liquid nitrogen. The sample was warmed at 34 °C/min, 4,600 °C/min and 6,600 °C/min. With EFS40, the survival was low regardless of the warming rate. With EFS30 and EFS20, survival was also low when the warming rate was low, but increased with higher warming rates, likely due to prevention of intracellular ice formation. When 1-cell embryos were vitrified with EFS20 and warmed rapidly, almost all of the embryos developed to blastocysts in vitro. Moreover, when vitrified 1-cell embryos were transferred to recipients at the 2-cell stage, 43% of them developed to term. In conclusion, we developed a vitrification method for 1-cell mouse embryos by rapid warming using cryotubes.     

The expression of Cav3.1 on T-type calcium channels of rats with subarachnoid hemorrhage

To study delayed cerebral vasospasm (DCVS) induced by subarachnoid hemorrhage (SAH), 60 healthy Sprague Dawley (SD) rats were randomly divided into 5 groups (12 rats in each group), namely sham operation group, blood injection model group, nimodipine group, flunarizine hydrochloride group, and normal group. Then, the physiological parameters were detected, and after the rats were killed under anesthesia, the degree of nerve injury, vasospasm as well as the therapeutic effect of drugs were evaluated by Western Blot (WB). Neurological impairment (NI), endothelial contraction and spasm were obvious in rats following blood injection. The expression of Cav3.1 on T-type calcium channels was significantly higher in the blood injection model group than in the sham operation group along with the normal group. Moreover, Cav3.1 mRNA was expressed in all groups. The Cav3.1 expression in blood injection model group and two drug groups were significantly higher than that in sham operation group and lower than that in blood injection model group. Vasospasm was improved in two drug groups, which indicated that calcium channel antagonists nimodipine and flunarizine hydrochloride had a certain therapeutic effect on DCVS in rats. The decrease in body weight and food intake of the two groups of rats treated with drugs decreased, and the delayed vasospasm was improved, but the expression of Cav3.1 was not changed significantly, indicating nimodipine and flunarizine hydrochloride had a therapeutic effect on delayed vasospasm in rats, but Cav3.1 expression on calcium channels was not affected.     

Foods of the south polar skua Catharacta maccormicki in the eastern Larsemann Hills,Princess Elizabeth Land,East Antarctica

Regurgitated pellets and food remains were collected near nest sites, and from a club site, of south polar skuas Catharacta maccormicki in the eastern Larsemann Hills, Princess Elizabeth Land, East Antarctica, during the skuas' presence in the area. The samples indicated that the snow petrel Pagodroma nivea, the most abundant seabird species breeding locally, formed the major dietary component, comprising some 66% of food items identified in pellets and 80% of the food remains obtained. Adélie penguins Pygoscelis adeliae (which do not breed in the Larsemann Hills), other seabirds, fish and marine foods were rarely found as remains or in pellets. However, refuse (meat, fish and vegetable remains) taken as food by skuas from nearby stations occurred in pellets at all sites and formed about 12% of the food remains collected and identified. In this study, foods taken by skuas were related both to the local breeding distribution of snow petrels, and to the possession of a feeding territory. Birds without feeding territories took somewhat fewer snow petrels and included more refuse from local stations in their diet, as did those at the club site. Future monitoring of the influence of anthropogenic foods (and inedible waste materials) on the species' ecology is considered important.     

Prorocentrum lima (Dinophyceae) in northeastern USA coastal waters: I. Abundance and distribution

The seasonal distribution of Prorocentrum lima within the epibiotic community associated with wild and cultured shellfish was studied at eight sites in northeastern USA coastal waters. This dinoflagellate produces toxins that can potentially accumulate in shellfish and lead to diarrhetic shellfish poisoning. Four sites in semi-sheltered environments provided sufficient data for an evaluation of population dynamics. The two southernmost populations (Point Judith Pond and Bluff Hill Cove in Rhode Island) displayed a double peak in abundance, one from March to June, with several thousands cells per g dry weight of collected epibiota, and a second one, sometimes minor, in autumn. At the two northern stations (New Meadows River and Clam Cove in Maine), P. lima populations also peaked twice but with a delay in timing. All four sites harbored filamentous seaweeds and aggregations of tube-forming or chains of diatoms, providing a favorable habitat for this epiphytic dinoflagellate.     

Inhibitory factors affecting the process of enhanced biological phosphorus removal (EBPR) – A mini-review

Enhanced biological phosphorus removal (EBPR) technology has been widely considered as a key strategy in preventing eutrophication and recognized as the advancing front of research in wastewater treatment. The key to keep its high efficiency in biological phosphorus removal is to optimize the operation and management of the system. Previous research in this field has undoubtedly improved understanding of the factors hindered overall efficiency of EBPR. However, it is obvious that much remains to be learnt. This paper attempts to review the fundamental understanding in factors inhibiting the stability and reliability of the EBPR systems in the state-of-the-art research. In view of modeling the EBPR systems, an appropriate extension of the current mechanistic models with these inhibitory factors is recommended in order to better simulate and predict the behavior of full-scale and lab-scale EBPR plants. From the perspectives of the further mechanistic and multi-factors study, the direction of denitrifying dephosphatation and granules/biofilms are also discussed. This comprehensive overview will not only help us to understand the overall mechanism of the EBPR process, but also benefit the researchers and engineers to consider all the possible factors affecting the process in the urban sewage treatment plants.     

余氯对水生生物的影响

氯是滨河、滨海企业冷却水常用的防治污损生物的处理剂。总结近年来国内外氯在企业冷却水中的应用、氯对浮游植物、浮游动物、贝类、鱼类等影响的研究成果,为制定冷却水余氯排放标准和水产养殖的合理布局提供了参考,针对我国大量滨海电厂即将建立的现状,分析了余氯研究在中国海域研究的不足,提出了氯对我国海洋生物影响不同层面的研究方向。认为氯对生物种群的毒性,氯对生物群落组成、结构和生态演替的影响,以及减轻或避免余氯污染的对策是需要进一步解决的科学问题。     

土壤样品中DNA提取方法的比较

对土壤样品中提取DNA方法的有效性进行了比较研究。如果以细胞有效裂解和DNA产率为标准,用冻融进行预处理再结合SDS和溶菌酶的化学裂解方法,是效果最佳的DNA抽提方法,细胞裂解率为82%,DNA产率达20.8μg/g。为了去除PCR抑制物,将DNA样品进一步用柱纯化,回收率为80%。纯化后的DNA样品可用于16SrDNA扩增及其他分子操作。