• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.169-174
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• 2006

To search of a new porcine pheromonal odorants, the comparative molecular similarity indices analysis(CoMSIA) between porcine odorant binding protein(pOBP) as receptor and ligands of green odorants 2-(Cyclohexyloxy)tetrahydrofurane derivatives as substrate molecule were conducted and disscused quantitatively. In the optimized CoMSIA model(I-AI) with chirality($I:\;C_{1'}(R),\;C_2(S)$) in substrate molecules and atom based fit alignment(AE) of the odorants the statistical PLS results showed the best predictability of the binding affinities based on the LOO cross-validated value ${r^2}_{cv.}\;(q^2=0.856)$ and non cross-validated conventional coefficient(${r^2}_{ncv.}=0.964)$). The structural distinctions of the highest active molecules were able to understand from the interaction between pOBP and green odorants in the contour maps with CoMSIA model.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.181-187
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• 2006

In recent years the number of patients waiting for organ transplantation has greatly outpaced the supply of human organs available, which leads to a renewed interest in pig-to-human xenotransplantation as an alternative. However, one of the biggest barriers in the xenotransplantation is presence of porcine endogenous retroviruses(PERV) that can infect human cells. In this study, to present a possible solution for this problem we tried to inhibit expression of PERVs using shRNAs(short hairpin RNA) at the level of RNA synthesis and virus release. The shRNA targeting the sequence of PERV A, B type was cloned into pSIREN-RetroQ vector under the control of polymerase-III U6-RNA gene promoter. Quantitative real-time PCR was performed to detect my alterations in mRNA production of PERV A, B targeted by the shRNA in each done. Depending on the target sequence of the shRNA, the transcription of PERV was decreased to as much as 4% and the number of progeny viruses was reduced to less than 1/200,000. Transgenic pigs producing such shRNAs may result in a highly reduced PERV expression in cells and organs, which is a prerequisite for safe xenotransplantations.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.104-108
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• 2007

The purpose of the study is to evaluate the clinical antagonistic effect of atipamezole(0.25 mg/kg, IM) in cats anesthetized with tiletamine-zolazepam ($Zoletil^{(R)}$, 10 mg/kg, IM) and medetomidine (0.05 mg/kg, IM). Twelve healthy 1 year old Korean mixed breed cats were used for this study. They were 4 males and 8 females. These cats were randomly assigned to two groups. One was control group ($Zoletil^{(R)}$ + medetomidine, ZM), and the other was treatment group ($Zoletil^{(R)}$ + medetomidine and antagonism by atipamezole, ZMA). All cats were examined 15 minutes before, 5, 25, 65 and 105 minutes after administration of tiletamine-zolazepam and medetomidine. Atipamezole was injected intramuscularly 20 minutes after ZM administation. Recovery time, heart rate, respiratory rate, total plasma protein and blood glucose were significantly different between ZM group and ZMA group (P<0.05). However, rectal temperature was not significantly different between ZM group and ZMA group. Two groups were able to induce sternal recumbency within 2 minutes and lateral recumbency within 4 minutes after the anesthetics injection. Mean sternal position time ($mean{\pm}SD$) was $174.0{\pm}44.6\;and\;116.2{\pm}27.3$ minutes, and mean standing position time was $210.8{\pm}45.6\;and\;154.2{\pm}21.1$ minutes in ZM and ZMA group, respectively. In these two groups, adverse effects during recovery time from anesthesia were not seen. As a result, the ZMA group had a faster recovery than the ZM group. Thus it was concluded that atipamezole could exert a useful reversal effect in cats anesthetized with medetomidine-tiletamine/zolazepam combination.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.153-163
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• 2002

In vitro matured porcine oocytes have very small volume of perivitellinspace (PVS). In these respect, the effect of sucrose and polybrene on the efficiency of gene transfer was investigated. As a gene (hGH) transfer vehicle, vesicular stomatitis virus glycoprotein pseudotyped retroviral vector (VSV-G) was used. Sucrose treatment has no detrimental effect on the rates of cleavage and resulted in the enlargement of PVS for the efficient introduction of retroviral vector stocks. Introduction rates of retrovirus in 0.5, 1, 2, 3 % sucrose treatment group were higher than that of the non-treatment group (39.3, 43.3, 35.7, 40.7 % vs. 8.3 %), respectively. In addition, we observed that sucrose pretreatment during injection procedure significantly reduce the frequency of polyspermy. In general, polybrene is a polycation essential for retrovirus transduction. The groups with the addition of 0.5, 5, 50$\mu\textrm{g}$/$m\ell$ polybrene exhibited a significant effect on gene transfer compared to that of the non-addition group (56.5, 50.0, 57.1 % vs. 34.6 %), respectively But, when the oocytes were co-injected with retrovirus and 50$\mu\textrm{g}$/$m\ell$ polybrene, the rates of cleavage and blastocyst development were 43.3 and 4.6%, respectively. This rates were lower than those of the non-addition group (70.0 and 17.3 %). In conclusion, sucrose pretreatment have increased efficiency of retroviral mediated gene transfer in porcine oocytes with no damage on in vitro fertilization and embryo development. In addition, sucrose pretreatment was beneficial in polyspermy inhibition. Presence of polybrene during microinjection showed a beneficial effect on the gene transfer in porcine oocytes, in low concentration. And these results will provide an useful tool for production of transgenic pigs by retroviral mediated gene transfer.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.245-255
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• 2004

In recent years, an increasing number of studies on pig in vitro maturation(IVM) and in vitro fertilization(IVF) have been separated. the wide range of new technologies, including that in applied molecular genetics, has increased this interest. the production of viable porcine embryos in vitro is a prerequisites for the successful production of transgenic pigs to date. The efficiency of IVM/IVF techniques in the porcine is lower than that obtained in other species such as cattle and mouse. The several problems are generally thought to be the cause of poor results: the low rate of MPN formation derived from inadequate IVM of oocytes, the high incidence of polyspermy after IVF and cell blocking at 4 cell during embryos culture. For there reasons overcoming, many studies have been conducted to improve in vitro embryo-genic competence of oocytes. In the last several years, many maturation culture media have been evaluated and various exogenous factors such as hormones and grows factors have been tested to improve the efficiency of porcine in vitro system. In the study several antioxidants have been examined to improve in vitro fertilization and development of porcine oocytes. In this study, several antioxidants were examined to determine the effects on the development of oocytes to the cleavage, morula and blastocyst stage when added at the maturation(IVM) or in vitro fertilization(IVF) or in vitro culture(IVC) of porcine embryos. Porcine oocytes were matured, fertilized and embryos were cultured in defind conditioned medium in vitro with or without supplementation with the antioxidents of cysteine, catalase and glutathione. 1. Significant improvement of blastocyst rate (27.2％ versus 15.4％, p<0.05) were achieved when catalase(500U/$m\ell$) were added to TCM-199 medium and morula rate(72.0％ versus 53.9％, p<0.05) were significantly higher when glutathione(1.0mM/$m\ell$) were added to TCM-199 medium than those of control. 2. In mTBM medium for oocytes fertilization, the addition of cysteine, catalase and glutathione had no positive effect on embryonic development. glutathione had no positive effect on embryonic development. In conclusion, this study shows that addition of catalase, gluththione during IVM improved the rate of porcine embryo development.

• Journal of Animal Science and Technology
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• v.46 no.4
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• pp.571-584
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• 2004

It is known widely that granulosa cell apoptosis leads follicular atresia and macrophages exert their effects directly and/or indirectly from the initiation to the completion of follicular atresia by phagocytosis of apoptotic bodies and secretion of various cytokines. However, the site of initiation, propagation routes and the elimination methods of apoptotic bodies, and the time and methods of penetration of macrophages into the follicles are not known completely. Using pig(Yorkshire-breed) ovary, immunohistochemical studies with TUNEL for apoptotic bodies and pig macrophage monoclonal antibody 4E9 for macrophages, and light and transmission electron microscopic observations were performed. In the pig, follicular atresia began with the granulosa cell apoptosis, and the apoptosis of theca intema cells occured at the same time. The apoptosis of granulosa cells initiated randomly within the granulosa cell layer and propagated rapidly into the whole layer. Ultrastructura1ly, apoptotic granulosa cells showed characteristic pyknotic and deformed nucleus and intracytoplasmic vesicles. Apoptotic bodies were eliminated by intact granulosa cells and macrophages. Intact granulosa cells ingested apoptotic bodies transiently, soon after they fell into the apoptosis. Finally, apoptotic bodies and degenerated oocyte were phagocytosed by macrophages. Macrophages entered the ovarian follicle at the time of initiation of granulosa cell apoptosis, and migrated with the progression of apoptosis. By elimination of theca cells, macrophages contributed the completion of follicular atresia These results will provide valuable informations on the study of the interrelation between macrophage and ovarian follicular atresia.

• Journal of Veterinary Clinics
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• v.26 no.6
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• pp.568-573
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• 2009

The purpose of this study is to evaluate the analgesic effects on the oral cavity in dogs which are treated with xylazine and electroacupuncture (EA). Furthermore, this study aims to find out its effects on glucose (GLU), serum alanine aminotransferase (ALT), and blood urea nitrogen (BUN) variation values, vital signs (rectal temperature, heart rate, respiratory rate) and pain responses to the noxious stimuli. Eight healthy dogs were randomly assigned to receive either xylazine or EA. Xylazine group dogs with weight of 3.6${\pm}$1.0 kg received 1.5 mg/kg of xylazine intramuscularly. EA group dogs with weight of 3.9${\pm}$1.0 kg received 1 volt (10-15 hz) for 5 minutes, and then 1-9 Volts (25-30 hz) for 60 minutes totally. The acupoints used were LI-3 (San Jian), LI-4 (He Gu) and ST-7 (Xia Guan). All dogs were examined before and 10, 25, 40, 55 and 120 minutes after administration of xylazine or EA. The mean rectal temperatures of the EA group were significantly higher than those of xylazine group after 25, 40 and 55 minutes (p < 0.05). The mean heart rates of the EA group were significantly higher than those of xylazine group after 10, 25, 40 and 55 minutes (p < 0.05). The mean respiratory rates of the EA group were significantly higher than those of xylazine group after 55 and 120 minutes (p < 0.05). The mean GLU concentration of the EA group were significantly lower than those of xylazine group after 55 and 120 minutes (p < 0.05). The sum of mean pain scores (SMPS) of the EA group were significantly higher than those of xylazine group after 10, 25 40 and 55 minutes (p < 0.05). In this study, the pain control of the EA group was shown to be better than that of the xylazine group. Also, there do not appear to be any negative physiologic effects associated with acupuncture-induced surgical analgesia. So, it was considered that these acupoints of EA analgesia might be useful for minor oral surgery in weak patients.

• Journal of Veterinary Clinics
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• v.27 no.5
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• pp.553-558
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• 2010

The purpose of this study was to investigate the effects of premedicated ascorbic acid and hepa-saline irrigation/aspiration on attenuation of ischemia-reperfusion (I/R) injury and recovery of renal function in canine nephrotomy model. In the canine model, nine mixed dogs were subjected to renal nephrotomy with premedicated ascorbic acid and hepa-saline irrigation-aspiration (treatment group 2), and only hepa-saline irrigation-aspiration (treatment group 1). The level of renal function and antioxidant enzymes after nephrotomy were measured. And the expression pattern of TNF-${\alpha}$ and INF-${\gamma}$ was examined in the renal tissue at $7^{th}$ day after nephrotomy. BUN and creatinine levels significantly decreased in the treatment group 1 and 2 compared to that of control group at the $3^{rd}$, 5th and $7^{th}$ day after reperfusion (p < 0.05). And, there was significant difference between treatment group 1 and 2 at the $3^{rd}$ day after reperfusion (p < 0.05). The activities of antioxidant enzymes in plasma was significantly increased in the treatment group 1 and 2 compared to that of control group at the $3^{rd}$, $5^{th}$ and $7^{th}$ day after reperfusion (p < 0.05). And, there was significant difference between treatment group 1 and 2 at the $3^{rd}$ day after reperfusion (p < 0.05). TNF-${\alpha}$ was decreased and INF-${\gamma}$ was increased in treatment groups. The result of this study suggested that irrigation-aspiration has effects on attenuation of renal ischemia-reperfusion injury, and the exogenous ascorbic acid has a role in the attenuation of renal ischemia-reperfusion injury and recovery of renal function in canine nephrotomy model.