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.     

Comparison of arylalkylamine N-acetyltransferase and melatonin receptor type 1B immunoreactivity between young adult and aged canine spinal cord

Melatonin affects diverse physiological functions through its receptor and plays an important role in the central nervous system. In the present study, we compared immunoreactivity patterns of arylalkylamine N-acetyltransferase (AANAT), an enzyme essential for melatonin synthesis, and melatonin receptor type 1B (MT2) in the spinal cord of young adult (2~3 years) and aged (10~12 years) beagle dogs using immunohistochemistry and Western blotting. AANAT-specific immunoreactivity was observed in the nuclei of spinal neurons, and was significantly increased in aged dog spinal neurons compared to young adult spinal neurons. MT2-specific immunoreactivity was found in the cytoplasm of spinal neurons, and was predominantly increased in the margin of the neuron cytoplasm in aged spinal cord compared to that in the young adult dogs. These increased levels of AANAT and MT2 immunoreactivity in aged spinal cord might be a feature of normal aging and associated with a feedback mechanism that compensates for decreased production of melatonin during aging.     

Increased humoral antibody response of foot-and-mouth disease virus vaccine in growing pigs pre-treated with poly-γ-glutamic acid

This study was conducted to determine if humoral antibody response of foot-and-mouth disease (FMD) vaccine improved in 8-week-old growing pigs born to well-vaccinated sows pre-treated with 60 mg of poly-γ-glutamic acid (γ-PGA) three days before vaccination. Antibody against FMD virus serotype O was measured 0, 2, 4 and 6 weeks post-vaccination, using a PrioCHECK FMDV type O ELISA kit. The results showed that positive antibody reactions against FMDV serotype O antigen among a component of the vaccine significantly increased in response to pre-injection with γ-PGA.     

Farmers' attitudes towards techniques for improving oestrus detection in dairy herds in South West England

Unidentified heats contribute to declining fertility rates in English dairy herds. Several techniques have been advocated to improve heat detection rates. Despite demonstrable technical efficacy and cost-effectiveness, uptake is low. A study in South West England used the Theory of Reasoned Action (TORA) to explore dairy farmers' attitudes and beliefs towards heat detection techniques. Few farmers were convinced that following prescribed observation times, milk progesterone testing and using pedometers would fit their system or improve on their current heat detection practices. Perceived difficulty of using a technique was not a constraint on adoption. Without promotion that addresses identified barriers and drivers to adoption, little change in current practice can be expected.     

Characteristics of Noodles and Bread Prepared from Double‐Null Partial Waxy Wheat

Double‐null partial waxy wheat (Triticum aestivum L.) flours were used for isolation of starch and preparation of white salted noodles and pan bread. Starch characteristics, textural properties of cooked noodles, and staling properties of bread during storage were determined and compared with those of wheat flours with regular amylose content. Starches isolated from double‐null partial waxy wheat flours contained 15.4–18.9% amylose and exhibited higher peak viscosity than starches of single‐null partial waxy and regular wheat flours, which contained 22.7–25.8% amylose. Despite higher protein content, double‐null partial waxy wheat flours, produced softer, more cohesive and less adhesive noodles than soft white wheat flours. With incorporation of partial waxy prime starches, noodles produced from reconstituted soft white wheat flours became softer, less adhesive, and more cohesive, indicating that partial waxy starches of low amylose content are responsible for the improvement of cooked white salted noodle texture. Partial waxy wheat flours with >15.1% protein produced bread of larger loaf volume and softer bread crumb even after storage than did the hard red spring wheat flour of 15.3% protein. Regardless of whether malt was used, bread baked from double‐null partial waxy wheat flours exhibited a slower firming rate during storage than bread baked from HRS wheat flour.     

Effects of three-dimensional spheroid culture on equine mesenchymal stem cell plasticity

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy fields. We optimize culture conditions of equine adipose tissue-derived MSCs (eAD-MSCs) for treatment of horse fractures. To investigate enhancing properties of three-dimensional (3D) culture system in eAD-MSCs, we performed various sized spheroid formation and determined changes in gene expression levels to obtain different sized spheroid for cell therapy. eAD-MSCs were successfully isolated from horse tailhead. Using hanging drop method, spheroid formation was generated for three days. Quantitative real-time PCR was performed to analyze gene expression. As results, expression levels of pluripotent markers were increased depending on spheroid size and the production of PGE₂ was increased in spheroid formation compared to that in monolayer. Ki-67 showed a remarkable increase in the spheroid formed with 2.0?×?10⁵ cells/drop as compared to that in the monolayer. Expression levels of angiogenesis-inducing factors such as VEGF, IL-6, IL-8, and IL-18 were significantly increased in spheroid formation compared to those in the monolayer. Expression levels of bone morphogenesis-inducing factors such as Cox-2 and TGF-β1 were also significantly increased in spheroid formation compared to those in the monolayer. Expression levels of osteocyte-specific markers such as RUNX2, osteocalcin, and differentiation potential were also significantly increased in spheroid formation compared to those in the monolayer. Therefore, spheroid formation of eAD-MSCs through the hanging drop method can increases the expression of angiogenesis-inducing and bone morphogenesis-inducing factors under optimal culture conditions.     

Injection molding of vertebral fixed cage implant

A vertebral cage is a hollow medical device which is used in spine surgery. By implanting the cage into the spine column, it is possible to restore disc and relieve pressure on the nerve roots. Most cages have been made of titanium alloys but they detract the biocompatibility. Currently PEEK (polyether ether ketone) is applied to various implants because it has good properties like heat resistance, chemical resistance, strength, and especially biocompatibility. A new shape of vertebral cage is designed and injection molding of PEEK is considered for production. Before injection molding of the cage, it is needed to evaluate process conditions and properties of the final product. Variables affecting the shrinkage of the cage are considered, e.g., injection time, packing pressure, mold temperature, and melt temperature. By using the numerical simulation program, MOLDFLOW, several cases are studied. Data files obtained by MOLDFLOW analysis are used for stress analysis with ABAQUS, and shrinkage and residual stress fields are predicted. With these results, optimum process conditions are determined.     

C-reactive protein, tumor necrosis factor α, and interleukin-6 in dogs with pyometra and SIRS

Objective: To determine the frequency of the systemic inflammatory response syndrome (SIRS) in canine pyometra and to evaluate the relationship between C‐reactive protein (CRP), tumor necrosis factor α (TNFα), interleukin‐6 (IL‐6), and SIRS. Design: Prospective clinical study. Setting: Veterinary teaching hospital. Animals: Fifty‐three clinical cases of canine pyometra and 19 healthy control bitches. Interventions: Upon admission to the veterinary hospital, history and physical examination findings, including previously defined clinical SIRS parameters, were documented. Blood samples were obtained for hematology and biochemical tests and for CRP, TNFα, and IL‐6 analysis. The diagnosis of pyometra was confirmed by histopathology of the uterus after ovariohysterectomy. After surgery, clinical SIRS parameters, length of hospitalization, and mortality were recorded. Measurements and main results: Pyometra dogs were grouped as SIRS positive (30/53; 57%) or SIRS negative (23/53; 43%). Logistic regression showed that CRP was the only parameter that significantly related to SIRS apart from the clinical criteria that define this syndrome. The mortality rate was low (2/53; 3.8%), and conclusions regarding association with SIRS could not be drawn. A positive SIRS status, high plasma CRP concentration, and high body temperature were variables that related to increased morbidity reflected by the length of hospitalization. Conclusions: SIRS was seen in 57% of canine pyometra cases and a positive SIRS status showed a positive association with prolonged hospitalization. The mortality rate was low (3.3%) among SIRS positive dogs, indicating that progression to multiple organ dysfunction syndrome (MODS) rarely occurs in surgically treated cases of pyometra. CRP was associated with SIRS and with prolonged hospitalization. Further studies of plasma CRP may be warranted in canine intensive care cases susceptible to development of SIRS and MODS.     

Risk analysis of Bovine Spongiform Encephalopathy in Korea

The occurrence of Bovine Spongiform Encephalopathy (BSE, so called mad cow diseases) that was first identified in England in 1986 was considered as being limited to only European countries, including England. However, the outbreak in Asia as well as North America since 2001 has amplified the fear that there isn't any nation in the world that is a safe area. In order to assess the risk of BSE outbreak in each country, the Office International des Epizooties (OIE) and EU have respectively established criteria, where OIE has set 5 levels and EU has set 4 levels. The Scientific Steering Committee (SSC) of the European Commission conducted a Geographical BSE Risk(GBR) assessment for 64 nations, such as the United States, etc., as of April 29, 2003. However, as of July 1, 2005, the duty of GBR assessment is expected to be transferred to a newly established body called EFSA (European Food Safety Authority, located in Parma, Italy). As Korea has not undergone a GBR assessment up to now, this study analyzed the risk of BSE outbreak in Korea by reviewing BSE prevention measures, etc., that have been put in place. This study shall be a barometer for estimating the GBR assessment level of Korea.     

工厂化繁育香菇胶囊菌种专用培养基配方研究

在已筛选出的代料香菇胶囊菌种专用配方LSC1和LSC5基础上,设计了7种香菇胶囊菌种培养基配方,以L26为供试菌株,观察测试了菌丝生长速度、瘤状突起物形成时间与数量、菌种成型及在愈伤培养时的特征以及菌种持水性等特征指标,筛选出了适宜于工厂化繁育香菇胶囊菌种的专用培养基配方LSCG2T和LSCG3,并在胶囊菌种工厂化繁育生产线中进行了应用试验,取得了良好效果。试验结果表明,工厂化繁育胶囊菌种的配方中粗细木屑的比例应提高到5∶1,配方中应添加以提供微量元素和基质物理性能改良剂的辅料。