Effect of 7,8-Dihydroxyflavone as an Antioxidant on In Vitro Maturation of Oocytes and Development of Parthenogenetic Embryos in Pigs

One of the factors that impairs in vitro produced porcine embryosis the oxidative stress that is mainly caused by the imbalance between reactiveoxygen species (ROS) generation and antioxidants activity, especially that ofglutathione (GSH). Here, we examined the effect of 7,8-dihydroxyflavone (7,8-DHF), akind of flavonoid antioxidant, on porcine oocyte maturation and its developmentalcompetence. Porcine oocytes were cultured in media supplemented with 0, 1, 5 and 10μM 7,8-DHF during both in vitro maturation (IVM) and invitro culture (IVC) after parthenogenetic activation. Maturation ofoocytes was evaluated based on first polar body (PB) extrusion and intracellular GSHlevel, and developmental competence was assessed through observing cleavage andblastocyst formation. In each step, the levels of intracellular GSH and ROS wereassessed by fluorescence intensity, and the apoptosis-related gene expression wasexamined using semiquantitative RT-PCR. The group treated with 1 μM 7,8-DHF duringIVM and IVC showed increased cytoplasmic maturation and reached the blastocysts stage(36.1%) at a higher rate than the other groups (24.7, 16.0 and 10.3% for 0, 5 and 10μM, P<0.05). In that group, the intracellular GSH level was significantlyincreased while ROS generation was significantly decreased after IVM and IVC(P<0.05). Moreover, it showed high expression of an anti-apoptotic gene(BCL2L1) and low expression of a pro-apoptotic gene(BAK1) (P<0.05). In conclusion, treatment with 1 μM 7,8-DHFduring IVM and IVC showed an anti-apoptotic effect by increasing intracellular GSHsynthesis and scavenging ROS and therefore improved the developmental competence ofporcine embryos.     

利用SAGE库挖掘有功能的假设基因

利用生物信息学方法,从人大动脉内皮细胞(HAECs)低氧处理的基因表达的系列分析(SAGE)中,挖掘出许多与低氧反应相关的假设基因。共预测到369个有功能的假设基因,其表达产物大部分与信号传导和基因的表达调控相关。选择了8个候选基因,RT—PCR验证其是否真正表达,其中7个为真基因。半定量RT—PCR表明,这些基因表达水平的变化在低氧处理的人脐静脉内皮细胞(HUVECs)中得到验证。结果表明：SAGE库中包含丰富的假设基因资源,可以用生物信息学方法从中鉴定与特定生理或病理过程相关的假设基因。     

Post-Death Cloning of Endangered Jeju Black Cattle (Korean Native Cattle): Fertility and Serum Chemistry in a Cloned Bull and Cow and Their Offspring

To preserve Jeju black cattle (JBC; endangered native Korean cattle), a pair of cattle, namely a post-death cloned JBC bull and cow, were produced by somatic cell nuclear transfer (SCNT) in a previous study. In the present study, we examined the in vitro fertilization and reproductive potentials of these post-death cloned animals. Sperm motility, in vitro fertilization and developmental capacity were examined in a post-death cloned bull (Heuk Oll Dolee) and an extinct nuclear donor bull (BK94-13). We assessed reproductive ability in another post-death cloned cow (Heuk Woo Sunee) using cloned sperm for artificial insemination (AI). There were no differences in sperm motility or developmental potential of in vitro fertilized embryos between the post-death cloned bull and its extinct nuclear donor bull; however, the embryo development ratio was slightly higher in the cloned sperm group than in the nuclear donor sperm group. After one attempt at AI, the post-death cloned JBC cow became pregnant, and gestation proceeded normally until day 287. From this post-death cloned sire and dam, a JBC male calf (Heuk Woo Dolee) was delivered naturally (weight, 25 kg). The genetic paternity/maternity of the cloned JBC bull and cow with regard to their offspring was confirmed using International Society for Animal Genetics standard microsatellite markers. Presently, Heuk Woo Dolee is 5 months of age and growing normally. In addition, there were no significant differences in blood chemistry among the post-death cloned JBC bull, the cow, their offspring and cattle bred by AI. This is the first report showing that a pair of cattle, namely, a post-death cloned JBC bull and cow, had normal fertility. Therefore, SCNT can be used effectively to increase the population of endangered JBC.     

IMAGING DIAGNOSIS—POSITIVE CONTRAST PERITONEOGRAPHIC FEATURES OF TRUE DIAPHRAGMATIC HERNIA

A true diaphragmatic hernia is a congenital diaphragmatic malformation that can appear identical to a peritoneopericardial diaphragmatic hernia (PPDH). True diaphragmatic hernias are rare in dogs. Herein we describe the use of positive contrast peritoneography for diagnosis of a true diaphragmatic hernia in two dogs.     

Contrast-enhanced ultrasonography to assess blood perfusion of skeletal muscles in normal dogs

This study evaluated perfusion of skeletal muscle using contrast enhanced ultrasonography in humerus, radius, femur and tibia in normal dogs. Contrast enhanced ultrasonography for each region was performed after injecting 0.5 mL and 1 mL of contrast medium (SonoVue) in every dog. Blood perfusion was assessed quantitatively by measuring the peak intensity, time to the peak intensity and area under the curve from the time–intensity curve. Vascularization in skeletal muscle was qualitatively graded with a score of 0–3 according to the number of vascular signals. A parabolic shape of time–intensity curve was observed from muscles in normal dogs, and time to the peak intensity, the peak intensity and area under the curve of each muscle were not significantly different according to the appendicular regions examined and the dosage of contrast agent administered. This study reports that feasibility of contrast enhanced ultrasonography for assessment of the muscular perfusion in canine appendicular regions.     

The effect of glycyrrhizin on rainbow trout, Oncorhynchus mykiss (Walbaum), leucocyte responses

Abstract. Treatment of rainbow trout macrophages with glycyrrhizin (GL), an aqueous extract of licorice, Glycyrrhiza glabra , enhanced their respiratory burst activity. Maximal effects were seen using concentrations of 10–100 μml⁻¹. GL also modulated trout lymphocytes, increasing proliferation responses to the mitogen phytohaemagglutinin some two-fold over a range of GL concentrations. In addition, GL elicited the release of a macrophage activating factor (MAF) from head kidney leucocytes, as assessed by the ability of generated supernatants to increase respiratory burst activity of target macrophages. MAF activity was most apparent using 100 μg ml⁻¹ GL to induce MAF release and a 48-h incubation period with the target macrophages. Finally, GL was shown to enhance the release of MAF in response to the mitogen concanavalin A. The possible use of GL as a stimulant of fish innate defences is discussed.     

不同浓度油酸对延边牛骨骼肌卫星细胞成脂转分化的影响

试验旨在探究油酸对延边牛骨骼肌卫星细胞成脂转分化的影响。试验设1个空白对照组（CON）和3个油酸（OA）诱导组：50、100、200 μmol/L油酸组（OAL、OAM、OAH）。油酸诱导96 h后,通过测定细胞大小及活力评估油酸对细胞的影响。通过油红O染色和测定甘油三酯来验证脂滴的形成,通过实时荧光定量PCR测定相关成肌成脂基因的表达水平来验证脂肪细胞的生成。结果显示,与对照组相比,添加油酸后,延边牛骨骼肌卫星细胞内有脂滴生成,并且脂滴形成量、甘油三酯累积量与油酸呈剂量依赖关系;实时荧光定量PCR测定结果表明,成肌相关因子Pax3、MyoD显著下调（P<0.05）,成脂相关因子C/EBPβ、PPARγ显著上调（P<0.05）,脂肪酸代谢相关因子SCD显著下调（P<0.05）,PLIN2基因显著上调（P<0.05）。综合上述试验结果,用油酸诱导处理延边牛骨骼肌卫星细胞可促进细胞的成脂转分化。     

Development of real-time onion disease monitoring system using image acquisition

In this study, real-time disease monitoring was conducted on onion which is the most representative crop in Republic of Korea, using an image acquisition system newly developed for the mobile measurement of phenotype. The purpose of this study was to improve the accuracy of prediction of disease and state variables by processing images acquired from monitoring. The image acquisition system was consisted of two parts, a motorized driving system and a PTZ (pan, tilt and zoom) camera to take images of the plants. The acquired images were processed as follows. Noise was removed through an image filter and RGB (red, green and blue) colors were converted to HSV (hue, saturation and value), which enabled thresholding of areas with different colors and properties for image binarization by comparing the color of onion leaf with ambient areas. Four objects with the most significant browning in the onion leaf to the naked eye were selected as the samples for data acquired. The thresholding method with image processing was found to be superior to the naked eye in identifying accurate disease areas. In addition, it was found that the incidence of disease was different in each disease area ratio. As a result, the use of image acquisition system in image processing analysis will enable more prompt detection of any changes in the onion and monitoring of disease outbreaks during the crop lifecycle.