Genetic relationships and evolution in Cucurbita as viewed with simple sequence repeat polymorphisms: the centrality of C. okeechobeensis

Genetic relationships among 88 accessions from nine of the dozen species of Cucurbita (Cucurbitaceae) were assessed from polymorphisms at 74 SSR (simple sequence repeat) loci originating from C. pepo and C. moschata, yielding a total of 315 alleles distributed among 17 linkage groups, with an average of 4.3 alleles per locus. Genetic distance (GD) values were calculated, a principal coordinate analysis conducted, and a dendrogram constructed. Average within-species genetic distance values ranged from 0.07 for C. ecuadorensis and C. ficifolia to 0.46 for C. pepo. Each species was clearly defined, as all mean within-species GD values were lower than the respective mean between-species GD values. C. okeechobeensis had the most central position in the genus Cucurbita, with the lowest average GD to the other species, 0.61. C. foetidissima, the only xerophytic species examined, was the most distant, with a mean GD of 0.73 to the other species. C. pepo and C. ficifolia were the most outlying of the mesophytic species. Mean across-species GDs generally corresponded with crossability. However, there were some outstandingly low GD values between particular accessions of Cucurbita pepo, the economically most important species, and disease-resistant wild species, particularly C. okeechobeensis but also C. foetidissima. The results suggest that more intensive search and collection of C. okeechobeensis populations would likely yield genotypes that are more compatible with C. pepo. Moreover, successful application of genetic resources in the genus Cucurbita might be facilitated by using GD values obtained from SSR polymorphisms as a guide in choosing parents for interspecific crossing.     

Maturation ability after transfer of freeze‐dried germinal vesicles from porcine oocytes

The purpose of this study was to examine whether freeze‐dried germinal vesicles (GV) can be matured in vitro after being injected into enucleated fresh oocytes in pigs as an alternative method for conservation of genetic resources. Although no reduction of the size of GV (p = .094), resveratrol treatment significantly enhanced the survival rates following GV transfer (GVT) (p < .001). Supplementation with 100 or 200 mmol/L trehalose in freeze‐drying medium significantly increased the proportions of GVs with intact nuclear membrane and DNA integrity compared with the control group. Following transfer of freeze‐dried GVs into enucleated fresh oocytes, the proportion of reconstructed oocytes reached the metaphase‐II stage (2.4% ± 1.4%) was significantly lower (p < .05) than that of the in vitro matured control group (83.2% ± 2.5%), it was comparable with the GVT control group (7.4% ± 2.7%). The rates of freeze‐dried GVs with intact nuclear membrane and DNA stored at ?20°C for 5 days were significantly higher (p < .05) than those at 4°C and room temperature. The rates of intact nuclear membrane and DNA in the freeze‐dried GV stored for 15 or 30 days at ?20, 4°C and RT were not significantly different. In conclusion, matured oocytes were produced derived from freeze‐dried GVs.     

Effect of vitrification at different meiotic stages on epigenetic characteristics of bovine oocytes and subsequently developing embryos

Vitrification by the Cryotop method is frequently used for bovine oocyte cryopreservation. Nevertheless, vitrified oocytes still have reduced developmental competency compared with fresh counterparts. The objective of this study was to compare the effect of vitrification either at the germinal vesicle (GV) stage or at the metaphase II (MII) stage on epigenetic characteristics of bovine oocytes and subsequently developing embryos. Our results demonstrated that vitrification of oocytes at each meiotic stage significantly reduced blastocyst development after in vitro fertilization (IVF). However, vitrification at the GV stage resulted in higher blastocyst development than did vitrification at the MII stage. Irrespective of the meiotic stage, oocyte vitrification did not affect 5-methylcytosine (5mC) immunostaining intensity in oocyte DNA. However, at both stages, it caused a similar reduction of 5mC levels in DNA of subsequently developing blastocysts. Oocyte vitrification had no effect on the intensity of H3K9me3 and acH3K9 immunostaining in oocytes and subsequent blastocysts. The results suggest that irrespective of meiotic stage, oocyte vitrification alters global methylation in resultant embryos although such alteration in the oocytes was not detected. Oocyte vitrification might not influence histone acetylation and methylation in oocytes and resultant embryos. Vitrification at the immature stage was more advantageous for blastocyst development than at the mature stage.     

Evaluation of isoflurane and sevoflurane vaporizers over a wide range of oxygen flow rates

OBJECTIVE: To examine the accuracy and precision of isoflurane and sevoflurane anesthetic vaporizers. SAMPLE POPULATION: 5 identical isoflurane and 5 identical sevoflurane vaporizers. PROCEDURES: Oxygen flow rates from 0.02 to 10 L/min were used with different vaporizer dial settings. Agent concentrations were measured at the common gas outlet by use of a refractometer. Accuracy was defined as the difference between measured agent concentrations, and dial settings were expressed as a percentage of the applied dial settings. Precision was defined as SD of the measured agent concentrations for each combination of dial setting and flow. RESULTS: Isoflurane values were generally greater than the dial settings. Accuracy of the isoflurane vaporizer was > 20% when 0.6% and 1% was dialed. Accuracy of the sevoflurane vaporizer was always within +/- 20% but decreased at 0.02 L/min flow and at combinations of high flow and high dial settings. Overall precision of the isoflurane vaporizer was better than that of the sevoflurane vaporizer. CONCLUSIONS AND CLINICAL RELEVANCE: A possible explanation for the inaccuracy of the isoflurane vaporizer may be that it was manufactured to be accurate with air but not oxygen, which must be accounted for when using the vaporizer with oxygen, especially with nonrebreathing systems. The sevoflurane vaporizer may not deliver accurate agent concentrations at high flow and high dial settings. Both vaporizers are suitable for clinical use with a wide range of oxygen flow rates if these precautions are properly addressed.     

Transgenic mice with a reduced core body temperature have an increased life span

Reduction of core body temperature has been proposed to contribute to the increased life span and the antiaging effects conferred by calorie restriction (CR). Validation of this hypothesis has been difficult in homeotherms, primarily due to a lack of experimental models. We report that transgenic mice engineered to overexpress the uncoupling protein 2 in hypocretin neurons (Hcrt-UCP2) have elevated hypothalamic temperature. The effects of local temperature elevation on the central thermostat resulted in a 0.3 degrees to 0.5 degrees C reduction of the core body temperature. Fed ad libitum, Hcrt-UCP2 transgenic mice had the same caloric intake as their wild-type littermates but had increased energy efficiency and a greater median life span (12% increase in males; 20% increase in females). Thus, modest, sustained reduction of core body temperature prolonged life span independent of altered diet or CR.     

Direct Evidence that the Number and Location of Cysteine Residues affect Glutenin Polymer Structure

In this study, the possible roles of three well-characterised model prolamins in the structure of the glutenin macropolymer were examined. Model prolamins were labelled with fluorescein isothiocyanate (FITC), and incorporated into the glutenin macropolymer of a base flour using a partial reduction-oxidation scheme. The effect of incorporation of the model prolamins on dough behaviour was determined by assessing differences in polymer size distribution, mixing properties, and distribution of the model prolamins in a dough after incorporation. Using this approach, the prolamins capable of forming inter-chain disulphide bonds were shown to be incorporated into the glutenin macroÍpolymer, while prolamins that were not capable of forming inter-chain disulphide bonds were retained as monomers. The distribution of fluorescently-labelled prolamins after their incorporation into the glutenin macropolymer of the dough was examined by confocal light scanning microscopy, in order to determine the possible roles of ω-gliadins and glutenin-like subunits with varied cysteine residue compositions in the structural organisation. The role of the model prolamins was a function of the disulphide-bonding capabilities of the polypeptides. Model ω-gliadins were retained as monomers and functioned as space fillers; model glutenin-like subunits containing a single cysteine residue incorporated into the glutenin macropolymer but functioned as chain terminators; and model glutenin-like subunits containing two cysteine residues incorporated into the glutenin macropolymer and acted as chain extenders.     

如何判断鱼卵的发育期?

亲爱的中国读者朋友、鲟鱼养殖生产者:·我是托马斯,匈牙利的独立鲟鱼专家,很多国家的人们称呼我“鲟鱼博士”。·我从事鲟鱼养殖、繁殖研究已经27年了。·我第一次成功人工繁殖小体鲟是在1979年。·我来中国传授鲟鱼繁殖技术也已经7年了。·我结识了许多中国的鲟鱼养殖生产者。·他们几乎都梦想着鲟鱼人工繁殖和加工鱼子酱,但许多技术问题在困扰着他们。·我的多年合作伙伴李融先生建议我借这个栏目,与中国朋友们分享我27年的经验。·今天,我们就来谈谈——     

如何正确投喂?——如果想做人工繁殖或者加工鱼子酱

亲爱的中国读者朋友、鲟鱼养殖生产者:l我是托马斯,匈牙利的独立鲟鱼专家,很多国家的人们称呼我“鲟鱼博士”。l我从事鲟鱼养殖、繁殖研究已经27年了。l我第一次成功人工繁殖小体鲟是在1979年。l我来中国传授鲟鱼繁殖技术也已经7年了。l我结识了许多中国的鲟鱼养殖生产者。l他们几乎都梦想着鲟鱼人工繁殖和加工鱼子酱,但许多技术问题在困扰着他们。l我的多年合作伙伴李融先生建议我借这个栏目,与中国朋友们分享我27年的经验。l今天,我们就来谈谈——李融:目前中国的鱼类养殖十分兴旺,也有许多类型的人工配合饲料供应。对于这些饲料在鲟鱼生…     

Excess polyspermy reduces the ability of porcine oocytes to promote male pronuclear formation after in vitro fertilization

Male pronucleus (MPN) formation is a very important physiological event during fertilization, which affects in vitro production of transferrable embryos. The aim of this study was to find out the correlation between the number of penetrated sperm and the occurrence of failure of MPN formation in porcine oocytes. In vitro matured porcine oocytes were fertilized in vitro with frozen epididymal sperm. Two different frozen sperm lots were tested in this study, which were different in terms of polyspermy rates. The numbers and the status of penetrated sperm in oocytes were evaluated 10 h after insemination. Under high polyspermy condition, the polyspermy rate was 83.5% with an average mean of 3.5 sperms per penetrated oocyte, whereas the percentage of polyspermy was 65.5% with an average mean of 2.4 sperms per penetrated oocyte under moderate polyspermic condition. Correlation analysis revealed a negative correlation between the number of penetrated sperm and their MPN formation percentage both in the sperm lot of high polyspermy (R = −0.560, p < 0.05) and in the sperm lot of moderate polyspermy (R = −0.405, p < 0.05) which suggests that penetration of excessive spermatozoa disables the oocyte cytoplasm to promote MPN formation.