• Journal of Life Science
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• v.25 no.8
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• pp.861-869
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• 2015

One of the domesticated species; the dog has been selectively bred for various aims by human. The dog has many breeds, which are artificially selected for specific behaviors and morphologies. Dogs contribute their life to human as working dogs for guide, rescue, detection or etc. Working dogs requires good personality, such as gentleness, robustness and patience for performing their special duty. Many studies have concentrated on finding genetic marker for selecting the high-quality working dog. In this study, we confirmed quantitative expression patterns of eight genes (ABAT; 4-Aminobutyrate Aminotransferase, PLCB1; Phospholipase C, Beta 1, SLC10A4; Solute Carrier Family 10, Member 4, WNT1; Wingless-Type MMTV Integration Site Family, Member 1, BARX2; BarH-Like Homeobox 2, NEUROD6; Neuronal Differentiation 6, SEPT9; Septin 9 and TBR1; T-Box, Brain, 1) among brains tissues from four dog breeds (Beagle, Sapsaree, Shepherd and Jindo), because these genes were expressed and have functions in brain mostly. Specially, BARX2, SEPT9, SLC10A4, TBR1 and WNT1 genes were highly expressed in Beagle and Jindo, and Sapsaree and German Shepherd were vice versa. The biological significance of total genes was estimated by database for annotation, visualization and integrated discovery (DAVID) to determine a different gene ontology (GO) class. In these analyses, we suppose to these eight genes could provide influential information for brain development, and intelligence of organisms. Taken together, these results could provide clues to discover biomarker related to functional traits in brain, and beneficial for selecting superior working dogs.

• Development and Reproduction
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• v.15 no.4
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• pp.349-357
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• 2011

Since nesfatin-1/NUCB2 involved in the control of appetite and energy metabolism was discovered for the first time in hypothalamus, many reports have shown its expression in various tissues. We also recently demonstrated that nesfatin-1/NUCB2 was expressed in the reproductive organs of mouse. However, no data exist on nesfatin-1/NUCB2 expression, regulation, and secretion in the uterus. Therefore, we examined the expression of nesfatin-1/NUCB2 in mouse uterus and the effects of PMSG and estrogen on its expression. NUCB2 mRNA expression in the uterus was determined by conventional and real-time PCR and nesfatin-1 protein expression was detected by western blotting. In immunohistochemistry staining, nesfatin-1 protein was localized at the epithelial cells of the uterine glands and endometrium. Nesfatin-1 protein binding sites were displayed at the epithelial cells of uterine glands and specific granulocytes including neutrophils. Additionally, to examine if the nesfatin-1/NUCB2 expression in the uterus is regulated by gonadotropin or estrogen, ovariectomized mice were treated with PMSG or $17{\beta}$-estradiol. The expression levels of NUCB2 mRNA in the uterus was significantly increased in the control mice after PMSG treatment, but not in the ovariectomized mice. In contrast, NUCB2 mRNA expression was dramatically increased in the ovariectomized mice after treatment with $17{\beta}$-estradiol. We report here for the first time that nesfatin-1/NUCB2 mRNA and protein express in the mouse uterus and its expression is regulated by estrogen secreted from the ovary, but not gonadotropin from the pituitary.

• Development and Reproduction
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• v.15 no.4
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• pp.373-383
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• 2011

Tributyltin (TBT) is one of endocrine disrupters which are known as having similar function to sex steroid hormone inducing apoptosis in various tissues of rodents. Recently, it has been reported that TBT induces apoptosis in thymus causing the decreased thymic function, but little is known about the mechanism. To elucidate the mechanism, three-week-old SD female rats were orally administrated with TBT 1, 10, and 25 mg per body weight (kg) and sesame oil as a control for 7 days. On day 8, the thymi were obtained and weighed, and then the number of thymocytes was counted. We also performed H&E staining, TUNEL assay, and Annexin V flow cytometric analysis to examine the apoptosis rates and the structure in the thymus. Next, we investigated the adipogenesis and apoptosis-related mRNA expression levels in the thymi by real-time PCR. The thymic weight and the number of thymocytes were decreased by TBT in a dose-dependent manner. As a result of the H&E staining, the boundary between cortical and medullary area was blurred in the thymi of TBT treated rats compared to those of controls. In the results of TUNEL assay and Annexin V flow cytometric analysis, apoptosis rates in the thymus were increased after TBT treatment. The expression levels of thymic epithelial cell marker genes such as EVA, KGF, AIRE, and IL-7 were significantly decreased in the thymi of TBT treated rats, but $PPAR{\gamma}$, aP2, PEPCK, and CD36 were significantly increased. The expression of $TNF{\alpha}$ and TNFR1 as apoptosis-related genes also was significantly increased after TBT treatment. The present study demonstrates that TBT can increase the expression of adipogenesis and apoptosis-related genes leading to apoptosis in the thymus. These results suggest that the increased adipogenesis of thymus by TBT exposure might induce apoptosis in the thymus resulting in a loss in thymic immune function.

• Development and Reproduction
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• v.14 no.4
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• pp.287-295
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• 2010

It was recently reported that nesfatin-1/NUCB2, which is secreted from the brain, controls appetite and energy metabolism. The purpose of this research was to confirm whether or not the protein and its binding site should have been expressed in the mouse reproductive organs and to know the possible effects of nesfatin-1 on the reproductive function. Using the ICR female mouse ovary and uterus, the expression of NUCB2 mRNA was confirmed with the conventional PCR and the relative amount of NUCB2 mRNA in the tissues was analyzed with real-time PCR. Immunohistochemical staining was performed using the nesfatin-1 antibody to investigate the nesfatin-1 protein expression and the biotin conjugated nesfatin-1 to confirm the binding site for nesfatin-1 in the ovary. Furthermore, in order to examine if the expression of NUCB2 mRNA in the ovary and uterus is affected by gonadotropin, its mRNA expression was analyzed after PMSG administration into mice. As a result, the expression level of NUCB2 mRNA in the ovary and the uterus was as much as the expression level in hypothalamus. As a result of the immunohistochemical staining, nesfatin-1 proteins were localized at the theca cells, the interstitial cells, and some of the luteal cells. However, the granulosa cells in the follicles did not stain. Interestingly, the oocytes in the some follicles were stained with nesfatin-1. On the other hand, nesfatin-1 protein binding sites were displayed at the theca cells and the interstitial cells near the tunica albuginea. After PMSG administration the expression level of NUCB2 mRNA was increased in the ovary and the uterus. These results demonstrate that for the first time the nesfatin-1 and its binding site were expressed in the ovary and NUCB2 mRNA expression was controlled by gonadotropin, suggesting an important role in the reproductive organs as a local regulator. Therefore, further study is needed to elucidate the functions of nesfatin-1 on the reproductive organs.

• Development and Reproduction
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• v.5 no.1
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• pp.47-52
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• 2001

Predetermination of sex in livestock of offpring is in great demand and is of critical importance to providing for the most efficient production of the animal ariculture. Such a sexing techlology would also enhance the economy of conventional artificial insemination(AI) and aid the porcine industry. The purpose of this study was to evaluate the efficiency of enriching X-bearing porcine sperm using discontinuous percoll gradients and PCR mefhod. Semen was collected from mature boars of proven fertility center (AI center KimHae). Sperm was leaded on the isotonic discontinuous percoll gradient and then it was centrifuged at 120 ${\times}$ g for 20 minutes. After centrifugation, sperm included in each fraction were recovered (7${\times}$10$^6$ sperms/ml) and then sperm genomic DNA was extractedfor the PCR. SRY gene was used to evaluate the ratio between X and Y sperm in the separated fractions. Ju viro ffrtilization wascarried out by adding the unseparated sperm (control) or separated (experimental poop) to the matured oocytes in TCM-199. Embryos for sex determination were obtained at 2 cell stage and then was used for SRY gene amplification. After centrifugation of discontinuous percoll gradient, the most motile sperm was obtained at 95% fiaction (94.4% ${\pm}$ 5.1%, p < 0.01). The PCR analysis evaluated that 30%, 50% and 65% fractions were Y sperm rich, whereas 80% and 95% fractions were X sperm rich. PCR analysis with each porcineembryo showed that 33.3% of control and 66.7% of experimental group were determined as female embryos. In conclusion, in vitro matured oocytes inseminated with sperms (95% fraction) prepared by percoll gradient centrifugation showed high fertilization rates and female embryos than control sperms.

• Korean Journal of Environmental Biology
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• v.35 no.2
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• pp.169-174
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• 2017

Acute and chronic toxicities of methyl ethyl ketone and methanol were investigated on Paronychiurus kimi (Collembola), for evaluating the potential effects of accidental exposures of these chemical substances on the terrestrial environments. This study was undertaken to establish a toxicity database for these chemical substances, which was required for the preparation of the response compensation and liability act for agricultural production and environmental damage. The 7-d acute toxicity and 28-d chronic toxicity were conducted using the OECD artificial soil spiked with varying, serially diluted concentrations of methyl ethyl ketone and methanol. Mortality was recorded after 7-d and 28-d of exposures, and the number of juveniles were determined after 28-d of exposure in the chronic toxicity test. In both assessments, methanol was more toxic than methyl ethyl ketone in terms of mortality ($LC_{50}$) and reproduction ($EC_{50}$). The 7-d $LC_{50}$ of methyl ethyl ketone and methanol were 762 and $2378mg\;kg^{-1}$ soil dry wt., respectively, and the 28-d $LC_{50}s$ were 6063 and $1857mg\;kg^{-1}$ soil dry wt., respectively. The 28-d $EC_{50}$ of methyl ethyl ketone and methanol were 265 and $602mg\;kg^{-1}$ soil dry wt., respectively. Comparison of results obtained in this study with literature data revealed that P. kimi was more sensitive to methanol than other soil invertebrates. However, given the high volatility of the chemicals tested in this study, further studies are necessary to improve the current test guideline, or to develop new test guidelines for an accurate assessment of chemicals that require toxicity databases for chemical accidents.

• Journal of Environmental Science International
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• v.10 no.4
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• pp.293-298
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• 2001

The toxicity values heavy metals were evaluated by immobilization and chronic reproduction impairment tests, using Daphnia magna. Acute tests were evaluated by the inhibition of their mobilization after 24hrs without food addition. The tests of reproductive impairment were investigated for 21 days by food addition and exchange or water. The effect of each concentration was assessed by Probit analysis and t-test. The results obtained from this study were as follows : 1) The change of pH and DO was not significant in the acute tests, while, in the reproductive tests, pH was increased by 0.3~1.4 and DO also increased. 2) The $E_iC_{50}$ values of immobilization to Daphnia magna in artificial fresh water were $0.030mg/{\ell}(Cu),\;0.054mg/{\ell}(Cd),\;0.12mg/{\ell};(Cr),\;0.74mg/{\ell}(Pb),\;3.4mg/ {\ell}(As)$ and the $NOE_iC$ values were $0.010mg/{\ell}(Cu),\;0.018mg/{\ell}(Cd),\;0.010mg/{\ell}(Cr),\;0.10mg/{\ell}(Pb),\;and\;$1.8mg/{\ell}(As)$. 3) The $E_rC_{50}$ values of reproductive impairment to Daphnia magna were $13.8\mu\textrm{g}/{\ell}(Cu),\;2.9\mu\textrm{g}/{\ell}(Cd),\;15.5\mu\textrm{g}/{\ell}(Cr),\;61.7\mu\textrm{g}/{\ell}(Pb),\;759\mu\textrm{g}/{\ell}(As)$, and $NOE_rC$ values were $0.95\mu\textrm{g}/{\ell}(Cu),\;$0.54\mu\textrm{g}/{\ell}(Cd),\;1.2\mu\textrm{g}/{\ell}(Cd),\;$7.4\mu\textrm{g}/{\ell}(Pb),\;110mu\textrm{g}/{\ell}(As)$. The results of tests using OECD artificial culture water were more sensitive than natural water for culturing. The presented data show that an artificial culture water is suitable in the experiment of bioassay for assessing the toxicity of marterials.

• Journal of the Korean Society of School Health
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• v.11 no.2
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• pp.229-241
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• 1998

Because sexual awareness is changing, I have tried to classify university students' attitudes toward sex. This study examined male and female university students and used a Q-method to measure these attitudes. Out of a total of 202, 33 statements were abstracted, and a P-sample of 1-7 parties consisting of 26 persons were studied. I Q-sorted the statements with a standard of 9 points for normal distribution and analyzed the data by means of a Principal Component Analysis with a Quan PC Program. According to the result of this research, university students' attitudes toward sex are divided into three types. Type 1 are called open-minded open: They may have a surgical abortion in case of an unexpected pregnancy and sexual intercourse occurs when a man and woman feel love for each other. They don't see sexual intercourse as a means reproduction nor virginal purity as unconditional. It is remarkable that these students regard sexual intercourse as a natural thing and sex as a basic desire of human beings. In short, they show an open-minded attitude toward sex. Type 2 are supporters of virginal purity; They regard virginal purity with great importance. They insist that both man and woman should be chaste but that sexual intercourse before marriage is unfavorable only to the woman. They do not believe in sexual intercourse without love and emphasize purity in sex education. Differen from type 1, they are very negative about surgical abortion. Type 3 experss love through sex. They belive that sexual intercourse makes a couple happy and a way to express their affection for each other. In other words, the relationship between man and woman is improved through sexual intercourse. They do not think of sexual intercourse in terms of childbirth.

• Korean Institute of Interior Design Journal
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• v.21 no.1
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• pp.78-85
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• 2012

The relevance between organisms and their external environment covers everything including humans, natural and artificial surroundings, regarding which academic and scientific understanding has continued. Relevant elements established by inter-dependence between humans and environment and the unity of life should be translated from the perspective of a whole, not of unit elements or reduction. That is, a space is formed by its own program and assumes sustainable relevance based on interactions between internal and external spaces, not building an independent system. The present study aims to present the feasibility of a potential mutant space formed by invisible arenas between individuals and evolutionary space formation based on an ecological paradigm Accordingly, this study suggested that evolutionary attributes as the major power source of biological changes could verify the virtual multiplicity of a new space formation, and that the potential form generation of hybrid mutant space of emergence and infinite formative capability could be supported. The suggestions made here will hopefully contribute to extending applicability of evolutionary space generation in the field of space design. To derive the potential mutant forms from biological space, a preliminary study was conducted regarding the characteristics of evolutionary form generation. For the purpose of this study, three evolutionary perspectives of reproduction, mutation (variation) and selection were taken. First, the theory of evolution was defined and characterized. Also, the relevance between the characteristics generated and hybrid mutant space was analyzed to consider relevant characteristics. The present study helped to understand that the hybrid mutant space had an evolutionary space structure based on a biological paradigm. It was also found that the mutant space structure built by mutant polymorphism assumed a systematic correlation between space and environment.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.7
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• pp.988-995
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• 2003

Physiological parameters were measured on six primiparous, non-pregnant Holstein cows prior to peak lactation over a 3-month summer season in southwestern Taiwan. The objectives were to characterize heat stress-induced change in functionality of mammary gland under natural climates of tropical summer and to establish physiological indices applicable to this environment in referring to this change. Environmental and physiological readings, milk and blood samples were taken at 15:00 h biweekly for totally five time points during the study. Climate readings showed that the afternoon humidex value reached the highest (53.5) around mid summer. Rectal temperature of cows taken simultaneously varied between $38.26^{\circ}C$ and $40.02^{\circ}C$ in parallel to humidex. Milk production declined drastically from 29.2 to 22.2 kg/d the first month entering summer but leveled up at end of the summer season suggesting effects exerted by heat stress rather than stages of lactation. Lactose content decreased linearly (p<0.05) with times in summer, from 4.69 to 4.38%. On the other hand, activity of N-acetylglucosaminidase (NAGase) in milk increased linearly to over two folds (p<0.05) during the same intervals. Elevations of fractional constituent of BSA in whey protein and serum cortisol level were also noticed in the course. Measurement of arteriovenous concentration (A-V) difference across the mammary gland demonstrated net uptake of glucose and net release of urea throughout the study period. The amount of urea released from mammary gland increased (p<0.05) progressively from 1.54 to 7.76 mg/dl during summer. It is concluded that gradual regression of mammary gland occurred along the humid tropical summer season. This regression is likely initiated through elevation of body temperature, which is irreversible above certain point. The increased release of urea from mammary gland during heat stress suggests its potential role as an early indicator of suboptimal mammary function.