Glutathione content of in vivo and in vitro matured canine oocytes collected from different reproductive stages

Glutathione (GSH) concentrations of oocytes are considered as an important marker of the cytoplasmic maturation. The present study was designed to compare GSH concentrations of in vivo and in vitro matured canine oocytes. In vivo matured oocytes were collected 72 hr after ovulation by flushing fallopian tubes after laparotomy. Ovaries were collected from bitches with different reproductive stages, and collected oocytes were divided into 2 groups according to the size viz. < 120 microm and > 120 microm in diameter and cultured for 72 hr in Tissue Culture Medium-199 supplemented with 10% FBS, 2.2 mg/ml sodium bicarbonate, 2.0 microg/ml estrogen, 0.5 microg/ml FSH, 0.03 IU/ml hCG, and 1% penicillin-streptomycin solution in the presence or absence of 50 microM beta-mercaptoethanol. GSH concentrations were determined by the dithionitrobenzoic acid-glutathione disulfide (DTNB-GSSG) reductase recycling assay. GSH concentrations of immature canine oocytes were 2.9 and 3.8, 3.5 and 6.8, and 3.1 and 6.5 pM/oocyte for < 120 microm and > 120 microm in diameter oocyte groups at anestrous, follicular and luteal stage, respectively (P<0.05). In vivo matured oocytes had significantly higher GSH concentrations compared with in vitro matured oocytes. The GSH content was 19.2 pM/oocyte for in vivo matured oocytes, while 4.1 to 8.1 and 5.7 to 13.2 pM/oocyte for in vitro matured oocytes cultured in the absence or presence of beta-mercaptoethanol, respectively (P<0.05). Presence of beta-mercaptoethanol increased GSH synthesis in canine oocytes cultured in vitro, and oocytes collected from follicular and luteal stage was superior to anestrus oocytes.     

鸡胚中沉积的尿酸盐对血凝试验的影响

在新城疫La Sota半成品检验过程中,发现接种后培养120 h的活胚,经过-20℃低温速冻1 h,有的鸡胚尿囊液中有尿酸盐产生,且出现的几率比较大,而经过2~8℃低温冷冻24 h的鸡胚,产生白色尿酸盐的几率相对小一些。凡是有尿酸盐产生的鸡胚,均没有血凝现象产生。但用含有尿酸盐的尿囊液接种鸡胚,继续增殖,会有病毒大量复制,试验说明有新城疫病毒存在的尿囊液中,若有尿酸盐沉淀,则血凝现象会被遮盖。     

Ameliorative Effects of Melatonin against Hydrogen Peroxide‐Induced Oxidative Stress on Boar Sperm Characteristics and Subsequent In Vitro Embryo Development

Melatonin, the major secretory product of the pineal gland, scavenges a variety of reactive oxygen and nitrogen species in vivo and in vitro, indicating that melatonin is a potent function as an antioxidant. The objective of this study was to investigate the effect of melatonin in the presence or absence of hydrogen peroxide (H₂O₂) on sperm characteristics (motility, viability, survival rate, membrane integrity, lipid peroxidation (LPO) and mitochondria activity) and also to examine the developmental rates to the blastocysts stage of porcine oocytes fertilized in vitro with semen treated with or without melatonin (100 nm ) in the presence or absence of H₂O₂ (250 μm ). The sperm were treated with melatonin in the presence or absence of H₂O₂ for 3, 6, 9 and 12 h at 37°C and then analysed for the sperm characteristics. The porcine embryos were produced by in vitro maturation and in vitro fertilization (IVM/IVF) using semen treated with or without melatonin (100 nm ) in the presence or absence of H₂O₂ (250 μm ) for 6 h. The semen characteristics, including motility, viability, survival rate, membrane integrity and mitochondria activity, were higher in the groups that were treated with melatonin in comparison to other groups, irrespective of incubation periods. Malondialdehyde levels in control, melatonin and melatonin + H₂O₂ groups were lower than H₂O₂ only group. A positive correlation was shown among motility, viability, survival rate and membrane integrity, but a negative correlation was observed between LPO and the other evaluation methods. The developmental rates to blastocysts of IVM/IVF porcine oocytes fertilized by semen treated with melatonin were significantly increased compared with any other groups, with the cell number of blastocysts shown to have a similar trend to the developmental rates. These results demonstrate that melatonin can improve the semen characteristics during in vitro storage and support the developmental ability of IVM/IVF embryos in pigs.     

Antioxidative Effects of Astaxanthin against Nitric Oxide‐Induced Oxidative Stress on Cell Viability and Gene Expression in Bovine Oviduct Epithelial Cell and the Developmental Competence of Bovine IVM/IVF Embryos

The aim of the present study was to elucidate the fundamental mechanism of bovine oviduct epithelial cell (BOEC) co‐culture on developmental capacity of bovine in vitro oocyte maturation/in vitro fertilization (IVM/IVF) embryos. We examined the effects of astaxanthin against nitric oxide‐induced oxidative stress on cell viability by MTT assay, lipid peroxidation (LPO) by using thiobarbituric acid (TBA) reaction for malondialdehyde (MDA) and the expression of antioxidant genes (CuZnSOD, MnSOD and Catalase) or apoptosis genes (Bcl‐2, Caspase‐3 and Bax) by RT‐PCR in BOEC. We also evaluated the developmental rates of bovine IVM/IVF embryos co‐cultured with BOEC pre‐treated with astaxanthin (500 μm ) in the presence or absence of sodium nitroprusside (SNP, 1000 μm ) for 24 h. Cell viability in BOEC treated with SNP (50–2000 μm ) lowered, while astaxanthin addition (50–500 μm ) increased it in a dose‐dependent manner. Cell viability in astaxanthin plus SNP (1000 μm ) gradually recovered according to the increase in astaxanthin additions (100–500 mm ). The LPO in astaxanthin group (50–500 μM) gradually decreased in a dose dependent manner and among SNP or astaxanthin plus SNP group, SNP alone and astaxanthin (50 μM) plus SNP shown a significant increase than other groups (p < 0.05). Expression of apoptosis or antioxidant genes was detected by RT‐PCR. Bcl‐2 and antioxidant genes were detected in astaxanthin or astaxanthin plus SNP group, and Caspase‐3 and Bax genes were only found in SNP group. When bovine IVM/IVF embryos were cultured for 6–7 days under co‐culture system such as BOEC treated with astaxanthin in the presence or absence of SNP, the developmental ability to blastocysts in 500 μm astaxanthin group was the highest of all groups. These results suggest that astaxanthin has a antioxidative effect on cell viability and LPO of BOEC, and development of bovine IVM/IVF embryos due to the induction of antioxidant genes and suppression of apoptosis genes.     

Physical properties of cryomilled rice starch

Cryomilling of rice starch was evaluated as a non-chemical way to modify starch structure and properties. Cryomilling in a liquid nitrogen bath (63–77.2 K) was done to Quest (10.80% amylose) and Pelde (20.75% amylose) rice starch at five different time frames (0, 15, 30, 45, and 60 min). The viscosity of the cryomilled rice starch decreased significantly (p < 0.05) with increasing milling duration, including peak viscosity, hot-paste viscosity, cold-paste viscosity, breakdown, and consistency. Increasing milling time significantly increased (p < 0.05) water solubility index and water absorption index. Infra-red spectroscopy and X-ray diffraction crystallography both showed that the crystallinity of the cryomilled starch decreased with increasing milling time. Differential scanning calorimetry (DSC) analyses showed that after 60 min cryomilling there was partial loss of crystallinity (86% for Quest and 91% for Pelde) of both cryomilled starches. The cryomilling process modified the rice starch by causing a loss of crystallinity, that reduced its pasting temperature and increased water absorption, and by fragmentation of starch (probably the amylopectin fraction) that reduced the viscosity and increased solubility.     

Comparison of In Vitro Developmental Competence of Cloned Caprine Embryos Using Donor Karyoplasts from Adult Bone Marrow Mesenchymal Stem Cells vs Ear Fibroblast Cells

The aim of this study was to produce cloned caprine embryos using either caprine bone marrow‐derived mesenchymal stem cells (MSCs) or ear fibroblast cells (EFCs) as donor karyoplasts. Caprine MSCs were isolated from male Boer goats of an average age of 1.5 years. To determine the pluripotency of MSCs, the cells were induced to differentiate into osteocytes, chondrocytes and adipocytes. Subsequently, MSCs were characterized through cell surface antigen profiles using specific markers, prior to their use as donor karyoplasts for nuclear transfer. No significant difference (p > 0.05) in fusion rates was observed between MSCs (87.7%) and EFCs (91.3%) used as donor karyoplasts. The cleavage rate of cloned embryos derived with MSCs (87.0%) was similar (p > 0.05) to those cloned using EFCs (84.4%). However, the in vitro development of MSCs‐derived cloned embryos (25.3%) to the blastocyst stage was significantly higher (p < 0.05) than those derived with EFCs (20.6%). In conclusion, MSCs could be reprogrammed by caprine oocytes, and production of cloned caprine embryos with MSCs improved their in vitro developmental competence, but not in their fusion and cleavage rate as compared to cloning using somatic cells such as EFCs.     

Enhanced intergrain tunneling magnetoresistance in half-metallic CrO2 films

Low-field tunneling magnetoresistance was observed in films of half-metallic CrO2 that were grown by high-pressure thermal decomposition of CrO3. High-temperature annealing treatments modified the intergrain barriers of the as-grown films through surface decomposition of CrO2 into insulating Cr2O3, which led to a threefold enhancement of the low-field magnetoresistance. This enhancement indicates the potential of this simple method to directly control the interface barrier characteristics that determine the magnetotransport properties.     

兔球虫与大肠杆菌病混合感染的诊治

阐述了兔球虫与大肠杆菌病混合感染的临床症状、实验室检验及防治方法,力求为科学防控两病提供有益的参考。     

Transendoscopic Nd:YAG Laser Surgery for Treatment of Epiglottal Entrapment and Dorsal Displacement of the Soft Palate in the Horse

Transendoscopic neodymium:yttrium-aluminum-garnet (Nd:YAG) laser was used to treat 12 standing horses with epiglottic entrapment (EE) or dorsal displacement of the soft palate (DDSP), or both. In four horses, transendoscopic laser staphylectomy was performed. The most common presenting complaints were respiratory stridor, cough, and exercise intolerance. Ten horses with EE healed without epiglottic complications; in one horse, partial adhesion of the aryepiglottic fold to one side of the epiglottis was corrected surgically through a laryngotomy incision. One horse with DDSP had no further signs, one continued to have continual DDSP, and two had induced DDSP. Transendoscopic Nd:YAG laser proved to be a feasible means of correcting EE and selected cases of DDSP.     

高效液相色谱法测定替米考星含量

建立了替米考星含量测定的HPLC方法。采用C18色谱柱(5μm,4.6 mm×250mm),乙腈+四氢呋喃+磷酸二丁胺为流动相,流速1.0 mL/m in,检测波长280 mn,以HPLC外标法测定替米考星含量。结果表明,实验测得的峰面积与对应的浓度呈良好的线性关系,其线性范围为0.25~0.75 mg/mL(r=0.999 9),平均回收率为99.81%。本法简便、准确、快速,是测定替米考星含量的良好方法。