• Journal of Periodontal and Implant Science
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• v.32 no.4
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• pp.865-873
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• 2002

The primary cause of tooth loss after 30 years of age is periodontal disease. Destruction of alveolar bone by periodontal disease is done by bone resorbing activity of osteoclasts. Understanding differentiation and activation mechanism of osteoclasts is essential for controling periodontal disease. The purpose of this study is to identify the possible effects of Vitamin D and cytokines affecting osteoclasts and its precursor cells. Four to six week-old mice were killed and humerus, radius, tibia and femur were removed aseptically and washed two times with Hank's solution containing penicillin-streptomycin and then soft tissue were removed. Bone marrow cells were collected by 22 gauge needle. Cells were cultured in Hank's solution containing 1 mg/ml type II collagenase, 0.05% trypsin, 41mM EDTA. Supernatant solution was removed 5 times after 15 minutes of digestion with above mentioned enzyme solution, and remained bone particles were maintained in alpha-MEM for 15 minutes and $4^{\circ}C$ temperature. Bone particles were agitated for 1 minute and supernatant solution containing osteoclast precursor cells were filtrated with cell stainer. These separated osteoclast precursor cells were dispensed with 100-mm culture dish by $1{\times}10^7$ cells unit and cultured in ${\alpha}$- MEM containing 20 ng/ml recombinant human M-CSF, 30 ng/ml recombinant human soluble osteoclast differentiation factor and 10% fetal calf serum for 2 and 7 days. Total RNA of osteoclast precursor cells were extracted using RNeasy kit. One ${\mu}g$ of total RNA was reverse transcribed in $42^{\circ}C$ for 30 minutes using SuperScriptII reverse transcriptase. Expression of transcribed receptors of each hormone and cytokine were traced with 1 ${\mu}l$ of cDNA solution by PCR amplification. Vitamin D receptor WAS found in cells cultured for 7 days. TNF-${\alpha}$ receptor was found in cells cultured for 2 days and amount of receptors were increased by 7 days. IL-1 type I receptor was not found in cells cultured 2 and 7 days. But, IL-1 receptor type II was found in cells cultured for 2 days. TGF-${\alpha},{\beta}$type I receptor was found in cells cultured 2 and 7 days, and amount of receptors were increased by 7 days of culture. These results implies Vitamin D and cytokines can affect osteoclasts directly, and affecting period in differentiation cycle of osteoclasts is different by Vitamin D and cytokines.

• Journal of Veterinary Clinics
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• v.20 no.3
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• pp.376-383
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• 2003

To establish the differential diagnosis and treatment method in bovine ovarian cysts, specially ovarian cysts with corpus luteum, serum progesterone concentration, rectal palpation and ultrasonography for measuring the cystic wall thickness and diameter of cyst and corpus luteum were investigated from 1,188 dairy cows with ovarian cysts. The plasma progesterone concentrations were 0.3$\pm$0.4 (mean$\pm$SD) ng/ml in 629 cows with follicular cysts, 3.7$\pm$1.1 ng/ml in 431 cows with luteal cysts, and 3.8$\pm$1.2 ng/ml in 128 cows with coexist of ovarian cysts and corpus luteum, respectively. The cystic wall thickness by ultrasonography were 1.6$\pm$0.4 mm in 629 cows with follicular cysts, 4.2$\pm$1.5 mm in 431 cows with luteal cysts, and 1.6$\pm$0.6 mm in 128 cows with coexist of ovarian cysts and corpus luteum, respectively. The days from initial treatment to insemination in follicular cysts were 28.1$\pm$6.9 days in treatment of GnRH alone, 15.9$\pm$2.9 days in combination of GnRH and dinoprost, and 15.1$\pm$3.1 days in combination of GnRH and cloprostenol. The percentages of cows conceived within 100 days after initial treatment were 61 %, 68% and 73% in treatment of GnRH alone, combination of GnRH and dinoprost, and combination of GnRH and cloprostenol, respectively. The days from initial treatment to insemination in luteal cysts were 3.8$\pm$0.6 days in treatment of dinoprost alone and 3.8$\pm$0.7 in cloprostenol alone. The percentages of cows conceived within 100 days after initial treatment were 69.5% and 68.5% in treatment of dinoprost and cloprostenol, respectively. The days from initial treatment to insemination in coexist of cysts and corpus luteum were 3.7$\pm$0.7 days in treatment of dinoprost alone and 3.8$\pm$0.6 in cloprostenol alone. The percentages of cows conceived within 100 days after initial treatment were 87% and 84% in treatment of dinoprost and cloprostenol, respectively. These results suggest that the best choice for treatment agents in ovarian cysts were combination of GnRH and PGF$_2$$\alpha$ in follicular cysts, and the PGF$_2$$\alpha$ in luteal cysts and in coexist of cysts and corpus luteum, respectively. In conclusion, it is suggest that ultrasonography is useful diagnostic tool for diagnosis and selection of treatment remedy in cystic ovaries of bovine.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.2
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• pp.133-138
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• 2006

Female phenotype of a 46,XY male may originates from male pseudohermaphroditism due to $17{\alpha}$-hydroxylase deficiency. Lack of cortisol increases adrenocorticotropic hormone (ACTH) and mineralocorticoid production, leading to low renin hypertention and hypokalemia. A 41-year-old phenotypic female presented primary amenorrhea and hypertension. In the hormonal profile, the levels of serum estradiol, testosterone, rennin, and cortisol were decreased and ACTH and deoxycorticosterone were increased. Laparoscopic bilateral gonadectomy was performed, and corticosteroid, antihypertensive drugs, and estrogen were administered. We report this case with a brief review of the literatures.

• Animal cells and systems
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• v.16 no.2
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• pp.127-134
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• 2012

We investigated the effects of fadrozol, an aromatase inhibitor (AI), and $17{\alpha}$-methyltestosterone (MT) on the induction of sex change in juvenile longtooth grouper $Epinephelus$ $bruneus$, via histological observation of gonads. Changes in the mRNA expression of GtH subunits (FSH-${\beta}$ and LH-${\beta}$) in the pituitary, and estradiol-$17{\beta}$ (E2) and 11-ketotestosterone (11-KT) levels in the blood were also surveyed after AI and MT treatment. Juvenile longtooth groupers ($113{\pm}17g\;body\;weight$; $16.2{\pm}1.2cm\;body\;length$) received intramuscular injections of AI at 3 (3-AI) and 5 (5-AI) mg/kg BWdoses and MT at a 5 mg/kg BW (5-MT) dose. At week 7 post-injection, 3-AI and 5-MT oocytes were degenerated, and gonads of the 5-AI group initiated spermatogenesis. At week 21 post-injection, 3-AI- and 5-MT-treated gonads contained spermatogonia and spermatocytes, while 5-AI treatment induced advanced stages of spermatogenesis. The serum E2 level showed no significant differences throughout the experimental period, whereas that of 11-KT was significantly elevated in the 5-AI group at weeks 7 and 21 post-injection. A significant increase in the expression of FSH-${\beta}$ mRNA was evident in the 5-AI group at week 21 post-injection. In contrast, LH-${\beta}$ mRNA expression did not significantly differ among groups during the experimental period. These results imply that sex change has two stages in the longtooth grouper. In the first stage, oocytes are degenerated by the stimulation by 11-KT, and in the second stage spermatogenesis occurs, owing to the co-effects of 11-KT and FSH-${\beta}$.

• Journal of Veterinary Clinics
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• v.19 no.4
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• pp.418-425
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• 2002

Estrus was induced in 13 anestrus Korea Jin-do bitches by intramuscular injection of pregnant mare serum gonadotropin (PMSG) in a dose of 500 lU once daily for ten consecutive days, followed by an additional single intraveneous injection of 1,000 lU of human chorionic gonadotropin (hCG) on the tenth day. Day-changes of vaginal epithelial cells during the hormone treatment were investigated in each experimental bitches and compared with the those of spontaneous estrus bitches. The first days of vulval bleeding and male acceptance after PMSG treatment were on Day 6.0$\pm$ 1.5 (mean$\pm$ SD) and Day 9.0$\pm$ 1.9, respectively. And in all of 13 bitches, vulval swelling and perineal reflex were shown. The mean durations of proestrus and estrus were 2.9$\pm$ 1.4 (mean$\pm$ SD, range ; 1-6) and 11.5: 1.7 (range ; 8-14) days, respectively, that is, duration of proestrus was significantly shorter than that of the spontneous estrous bitches but duration of estrus was longer than that of the spontaneous estrous bitches. Characteristic features of vaginal cytology during the estrous cycle were the high proportions of large intermediate cell, superficial cell, anuclear cell and erythrocyte in proestrus, superficial cell and anuclear cell in estrus and parabasal cell, small intermediate, large intermediate cell, and leukocyte in diestrus, respectively. The comification index (Cl) was significantly high proportion in proestrus and estrus, when Day 0 was timed from the first day of male acceptance, the Cl was first increased above 80% on Day 0 and maintained above 80% until Day 0 to Day 5 during 6 days and showed a peak on Day 2. Also it was maintained above 90% until Day 2 to Day 3 during 2 days. These results indicated that all 13 ekperimental bitches showed positive estrus detection by the estrus behavior and vaginal smear test after treated with PMSG and hCC. It suggested that vaginal cytology was used to estimate the optimal mating and ovulation time, in consideration of the day when the Cl was maintained above 80% in estrus-induced Korea Jin-do bitches.

• Journal of Veterinary Clinics
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• v.24 no.1
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• pp.1-4
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• 2007

Despite the risk of high altitude disease (HAD), raising cattle at higher altitudes does have advantages such as beef quality due to minimal chance of infection and stress. The ideal situation is to determine the optimum altitude suitable for raising cattle while at the same time minimizing the risk of HAD. Therefore, in this study, we documented the health status of three groups of steers, raised at three different altitudes (200m, 400m and 800m) in Gangwon province, using routine hematological, biochemical and hormonal tests. The red blood cell counts and hemoglobin concentration were highest in the 800m group, and lowest in the 200m group, possibly due to hypoxia induced myeloid and erythroid stimulation. The mean concentration of AST, BUN and serum cortisol were lowest in the 800m. These findings suggest the stress factor will contribute the general health status of animal and indicate a difference in the Hanwoo groups raised at 800m compared to 200m, where the cattle raised at the higher altitude exhibit better health status compared to the lower altitude, possibly due to less stressful environment in the higher altitude.

• Journal of Ginseng Research
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• v.41 no.4
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• pp.578-588
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• 2017

Background: Elevated testicular temperature disrupts spermatogenesis and causes infertility. In the present study, the protective effect of enzymatically biotransformed Panax ginseng Meyer by pectinase (GINST) against chronic intermittent heat stress-induced testicular damage in rats was investigated. Methods: Male Sprague-Dawley rats (4 wk old, 60-70 g) were divided into four groups: normal control (NC), heat-stress control (HC), heat-stress plus GINST-100 mg/kg (HG100), and heat-stress plus GINST-200 mg/kg (HG200) treatment groups. Each dose of GINST (100 mg/kg and 200 mg/kg) was mixed separately with a regular pellet diet and was administered orally for 24 wk. For inducing heat stress, rats in the NC group were maintained at $25^{\circ}C$, whereas rats in the HC, HG100, and HG200 groups were exposed to $32{\pm}1^{\circ}C$ for 2 h daily for 6 mo. At week 25, the testes and serum from each animal were analyzed for various parameters. Results: Significant (p < 0.01) changes in the sperm kinematic values and blood chemistry panels were observed in the HC group. Furthermore, spermatogenesis-related molecules, sex hormone receptors, and selected antioxidant enzyme expression levels were also altered in the HC group compared to those in the NC group. GINST (HS100 and HS200) administration significantly (p < 0.05) restored these changes when compared with the HC group. For most of the parameters tested, the HG200 group exhibited potent effects compared with those exhibited by the HG100 group. Conclusion: GINST may be categorized as an important medicinal herb and a potential therapeutic for the treatment of male subfertility or infertility caused by hyperthermia.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.293-300
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• 1997

Supplementation of maturation medium with additional granulosa cells has beneficial effect on in vitro maturation of bovine follicular oocytes and their subsequent cleavage and development in vitro. However, maturation system using granulosa cells have some disadvantages that collection of granulosa cells is cumbersome and metabolic activity of the cells is variable according to ovarian cycle or follicular size. We hypothesized that bovine immsture oocytes matured without granulosa cell coculture can fertilize and develop normally if the medium volume per oocyte is reduced during in vitro maturation. Immature oocytes were matured for 24 hours in a TCM199 containing 10% fetal calf serum, anterior pitultary hormone (0.02 AU /ml Antrinⓡ) and estradiol with or without granulosa cells in vitro. In Group 1, 35 to 40 oocytes were matured in a well of 4-well plastic dish containing 500 $\mu$l of maturation medium and granulosa cells, and 9 to 10 oocytes were matured in a 50-$\mu$l drop of maturation medium without granulosa cells in Group 2. After maturation, oocytes were coincubated with sperm for 30 hours in a modified Tyrode's medium (IVF). Inseminated oocytes were cultured in a microdrop (30 $\mu$l) of a synthetic oviduct fluld medium (SOFM) containing BSA, Minimum Essential Medium essential and non-essential amino acids for 9 days. As a preliminary experiment, we investigated the beneficial effect of granulosa cells during maturation on subsequent cleavage and development using the same type of culturedishes (4-well dish). Granulosa cells could not increase embryo cleavage after fertilization but significantly improved (p<0.05) embryo development to expanding blastocyst (Table1 and 2). In Group 1, 68 and 80% of inseminated oocytes have cleaved at 30 hours and 2 days after IVF, respectively, which is similar (p>0.05) to the result of Group 2 (69% at 30 hours and 78% at 2 days after IVF). The oocytes in Group 2 showed 21 and 11% of developmental rates to expanding and hatching blastocysts, respectively, which was not significantly different (p>0.05) from those (20 and 10%, respectively) of oocytes in Group 1. In conclusion, it has been clarified that a microdrop culture system without granulosa cells for in vitro maturation can support bovine embryonic development to blastocyst in vitro as readily as a granulosa cell coculture system.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.95-104
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• 2002

This study was performed during the four seasons for the production of transgenic pigs containing the human erythropoietin(hEPO) transgene. Purebred Landrace gilts and sows approximately 8∼15 months of age (n=42) were used fur the collection of 1-cell zygotes for DNA microinjection and transfer. Retrospectively, estrus synchronization and superovulation schemes were evaluated to assess practicality for zygote collection. Synchronization and superovulation procedures were used that cyclic gilts were synchronized with 20mg altrenogest (ALT) per day for 9days after PG600 administration followed by superovulation with 1500IU pregnant mares serum gonadotropin (PMSG) and 500IU human chorionic gonadotrophin (hCG). Preparation of recombinant gene for microinjection is mice whey acidic protein promoter (mWAP) linked to human erythropoietin (hEPO) gene. After hormone treatment, 650 embryos were collected from 23 donors and 83.1% (540/650) embryos were in 1-cell stage which can be visualized the pronuclei for DNA microinjection. A total of 543 DNA microinjected embryos fiom donors were transferred to 19 synchronized recipients, seven of them maintained pregnancy and delivered 47 piglets. One of the 47 offsprings were determined to have transgene by PCR analysis. The overall rate of transgenic production was 2.13% (tansgenic/offspring). This study provides the success and useful information regarding production of transgenic pig for bioreactor research.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.87-94
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• 2002

This study was performed during the four seasons for the production of transgenic pigs containing the Cellulase Digest Gene. Purebred Landrace gilts and sows approximately 8∼15 months of age (n=126) were used for the collection of 1-cell zygotes for DNA microinjection and transfer. Retrospectively, estrus synchronization and superovulation schemes were evaluated to assess practicality fur zygote collection. Synchronization and superovulation procedures were used that cyclic gilts were synchronized with 20mg altrenogest (ALT) per day for 9 days after PG600 administration followed by superovulation with 1000 IU pregnant mares serum gonadotropin (PMSG) and 750IU human chorionic gonadotrophin (hCG). The cellulase digestion gene for microinjection is rat elasterase promoter (rEl) linked to CelD gene. After hormone treatment, 1,422 embryos were collected from 91 donors and 95.6% (1,359/1,422) embryos were in 1-cell stage which can be visualized the pronuclei for DNA microinjection. A total of 725 DNA microinjected embryos transferred into 35 recipients and produced 65 piglets from 13 litters. Pregnancy rate according to the number of transferred embryos to recipients was higher the group which received 21 to 24 embryos (50.0%) than other groups 20.0% in less and 33.3% in more. A tail tissue was collected from 65 piglets for biopsy. PCR screening was performed on each DNA sample using two separate sets of primers specific for the 5'- and 3'-flanking region of the rEl-CelD gene. Five of the 65 piglets (7.69%) were positive for the transgene. This study provide useful information regarding production of transgenic pig for bioreactor research.