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.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

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.     

The Comet assay for detection of DNA damage in canine sperm

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno‐, terato‐ or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A—median: 31.55%, interquartile range (IQR): 30.18–38.01; group B—median: 0.90%, IQR: 0.65–1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = ?.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non‐normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.