• Applied Microscopy
• /
• v.7 no.1
• /
• pp.21-43
• /
• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

• Journal of Embryo Transfer
• /
• v.21 no.3
• /
• pp.255-262
• /
• 2006

The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.

• Journal of Veterinary Clinics
• /
• v.27 no.5
• /
• pp.522-532
• /
• 2010

The aims of the present study were to characterize the effect of glucocorticosteroids (GCs) on the normal canine skin and to evaluate the effect of a lipid mixture (LM), containing cholesterol, pseudoceramide, and free fatty acid, on the steroid-induced damaged skin of dogs. Five beagles were involved and the skin of the back of each dog was topically applied with four kinds of GCs twice daily for 28 days. LM was applied after that period of GCs application. Transepidermal water loss (TEWL), skin hydration, and skin pH were assessed during experimental periods and histopathological evaluation was performed. TEWL was significantly increased, with a maximum increase obtained on day 28 (p < 0.01). Skin pH was significantly decreased, with a maximum decrease obtained on day 28 (p < 0.01). Skin surface hydration was significantly increased on day 3, but values of skin hydration were progressively decreased and finally reached those of baseline. In histology, as results of steroid application, losses of keratin layers in the stratum corneum and edematous changes in the upper parts of dermis, and consequently, thickness of the epidermis and the stratum corneum were decreased. In addition, the numbers of hair follicles were markedly decreased in steroid control as compared to intact control. However, these skin atrophic changes were markedly inhibited by treatment of LM as compared with steroid control in the present study. Moreover, all biophysical parameters were reached to the baseline after LM treatment. These results showed that the topically applied GCs induced skin barrier impairment and a LM should be effective on repair of disturbed skin barrier function in dogs. Therefore, it is concluded that a LM tested in the present study is expected to treat the steroid-induced skin damages.

• Clinical and Experimental Reproductive Medicine
• /
• v.13 no.1
• /
• pp.11-19
• /
• 1986

Follicular flxid (FF) and their matched oocytes were obtained from 58 follicles of 27 women who underwent an in vitro fertilization (IVF) procedure with ovarian hyperstimulation by clomiphene citrate(n=8), hMG(n=9),FSH/hMG(n=10). Follicular aspiration was performed 36 hours after human chorionic gonadotropin administration. The concentcation of androstendione (ADD), testosterone (T) was correlated with hyperstimulation regimens, the morphology of the oocyte-corona-cumulus complex (OCCC), oocyte fertilization, and the incidence of pregnancy after embryo transfer. The results were as follows. 1. According to hyperstimulation regimens, there was no significant differance in FF ADD and T concentrations of the similar morphology of OCCC. 2. In clomiphene-treated and FSH/hMG-treated cycles, FF ADD concentrations of preovulatory oocytes were 43.09${\pm}$9.53 ng/ml and 59.46${\pm}$9.09 ng/ml, those of immature occytes were 96.98${\pm}$16.55 ng/ml and 116.13${\pm}$36.81 ng/ml, those of atretic oocytes were 246.5 ${\pm}$9.25 ng/ml and 634.25${\pm}$9.25 ng/ml respectively, reflecting the significant relationship between FF ADD level and morphologic maturity of OCCC (p<0.05). But in hMG-treated cycles, such relationship was not found (p>0.1). In clomiphene-treated and FSH/hMG-treated cycles, FF T concentrations of preovulatory oocytes were 11.37${\pm}$2.38 ng/ml and 11.68${\pm}$1.73 ng/ml respectively which were significantly lower than those of atretic oocytes (25.1${\pm}$7.50 ng/ml and 23.25${\pm}$0.95 ng/ml respectively) (p<0.05). But in all cycles, FF T concentrations of immature oocytes were not significantly different from those of preovulatory oocytes, artetic oocytes (p>0.1). 3. In hMG-treated and FSH/hMG-treated cycles, FF ADD concentrations of fertilized oocytes were 32.43${\pm}$4.09 ng/ml and 42.61${\pm}$4.82 ng/ml respectively which were significantly lower than those of non-fertilized oocytes (72.18${\pm}$17.31 ng/ml and 108.09${\pm}$17.32 ng/ml respectively) (p<0.05), but in clomiphene-treated cycles there was no significant difference (p>0.1). In FSH/hMG-treated cycles, FF T concentration of fertilized oocytes was 7.33${\pm}$1.06 ng/ml which was significantly lower than that of non-fertilized oocytes (20.3${\pm}$6.21 ng/ml) (p>0.02), but in clomiphne-treated and hMG-treated cycles there was no significant difference (p>0.1). 4. In all cycles FF ADD and T concentrations did not correlated with the success of pregnancy after embryo transfer. Above results suggested that FF ADD and T may play an important role in oocyte maturation and fertilization, but their relationship with the success of psegnancy was not found.

• Journal of Embryo Transfer
• /
• v.29 no.3
• /
• pp.265-271
• /
• 2014

Implementation of smart embryo technologies in cattle e.g. ovum pick-up followed by in vitro embryo production (OPU-IVP). Seasonal variation is important factor for follicular growth, oocytes quality, quantity and developmental competence. Therefore the aim of present study was carried out to investigated whether the seasons (hot and cool) effect on follicular development, oocyte recovery and subsequent embryo development. Follicular oocytes were aspirated from Korean native cows (Hanwoo) by the ovum pick-up (OPU) method, which was performed 24 times during two different seasons, the hot (July to September) and cool (October to December), from OPU donors. The recovered oocytes were classified according to morphological categories and used for in vitro embryo production (IVEP). The mean number of total follicles was significantly higher (p<0.05) during the hot season ($18.32{\pm}2.26$) compared to cool season ($15.41{\pm}3.34$). Furthermore, seasons did not significantly effect on the number of oocytes recovered (hot season: 41.16% vs. cool season: 46.14%). However, the average number of Grade A oocytes was significantly greater during hot ($1.75{\pm}1.86$) season compared to the cool season ($1.00{\pm}1.46$), but there was no significant difference of other grades oocytes. The cleavage rate (hot: 66.67% vs. cool: 63.3%) and embryo development (hot: 58.95% vs. cool: 56.97%) did not differ significantly between the seasons. In conclusion, the results of present study suggest that the season (hot and cool) does not have effects on the oocyte recovery and embryo developmental competence of in vitro cultured embryos.

• Korean Journal of Animal Reproduction
• /
• v.6 no.1
• /
• pp.19-30
• /
• 1982

1. The cows slaughtered at age of 3, 4, 6, 7, 8, and 9 years old were 1.5, 1.5, 15.0, 62.5 and 4.4% respectively. 2. The cows slaughtered at 351-450kg and more than 500kg were 60 and 28% respectively. 3. Best, very good, good and bad cows in nutritional condition were 1.6, 25.8, 62.9, and 9.7% respectively. Among the six cows which were bad nutrition, the two were with severe endometritis, the three were normal in genital function and one was on 70 days of pregnancy. 4. Holstein cows(55.2%) showed higher reproductive failure than the Korean cows(33.3%). 5. The slaughted ratio of the Korean cattle and Holstein cows was 36 and 64% respectively. 6. Pregnant cows were about 16% among the slaughtered one. 7. Reproductive failures were composed of 46% in uterus, 32% in ovaries, 8% in udder, 6% in oviduct, 4% in cervix of uterine, 2% in vagina and 2% inmummified fetus. 8. Forty six percentages of uterine diseases were as follows; horn, 13%, body of uterus, 32% and ovary diseases were 32%, that is, 12% of ovary atrophy, 8% of ovarycyst and 6% of lutealcyst. 9. The cows of reproductive failures were commonly infected with 1.6 kinds of diseases. 10. According to classification, six type of ovaries were as follows; normal, 58%, ovary-cyst, 11%, luteum cyst, 4%, coexistence of follicles and corpus luteum, 16%, weak function of ovaries, 10% and ovarian atrophy, 1%. 11. Major axis, minor axis and thickness of right ovary were larger than those of left one both in Korean cattle and Holstein cows. Holstein cow had generally larger size of ovary than these of the Korean cattle.. 12. The left and right oviducts showed no difference in length, but Holstein had longer oviduct than Korean cow. 13. There was no difference in the length of uterine horn between right and left in the Korean cows, but the right was longer than the left in Holstein cows. 14. Holstein had longer horn and body of uterine than the Korean cows. 15. The weight of right ovary was heavier than that of left in both breeds, but there was no differences in weight of left ovary between two breeds and right ovary of Holstein breed was heavier than that of the Korean cow. 16. The weight of right oviduct and uterine born was heavier than that of the left, and Holstein had heavier oviducts and uterine horns than the Korean cows. 17. Holstein had heavier uterine body and cervix of uterine than the Korean cows. 18. The length of reproductive systems of Korean cow is as follows; Major and minor diameter and thickness ofovary are 3.6${\pm}$0.7, 2.3${\pm}$0.4 and 1.6${\pm}$1.4 cm in left and 3.7${\pm}$0.6, 2.5${\pm}$0.5 and 1.8${\pm}$0.5 cm in right. Oviduct is 28.4${\pm}$3.1 cm in left and 27.8${\pm}$3.3 cm in right. Uterine horn is 27.4${\pm}$4.5 cm in left and 27.7${\pm}$4.9 cm in right. Uterine body and cervix are 3.4${\pm}$1.1 and 6.5${\pm}$1.7 cm. 19. The length of female reproductive systems ofHolstein cow is as follows; Major and minor diameter and thickness of ovary are 3.9${\pm}$1.3, 2.3${\pm}$0.5, and 1.5${\pm}$0.6 cm in left and 4.0${\pm}$0.8, 2.8${\pm}$0.6 and 1.8${\pm}$0.6 cm in right. Oviduct is 29.4${\pm}$4.2 cm in left and 29.3${\pm}$4.1 cm in right. Uterine horn is 30.2${\pm}$7.4 cm in left and 32.6${\pm}$8.4 cm in right. Uterine body and cervix are 4.5${\pm}$2.5 and 7.8${\pm}$2.9 cm. 20. The weight of reproductive systems of Korean cow is as follows; Ovary is 8.4${\pm}$4.1 g in left and 9.3${\pm}$3.6g in right. Oviduct is 1.5${\pm}$0.5 g in left and 1.6${\pm}$0.5 g in right. Uterine horn is 109${\pm}$27 g left and 118${\pm}$32 g in right. Uterine body and cervix are 30.4${\pm}$14.1 and 76.7${\pm}$38.4g. 21. The weight of reproductive systems of Holstein cow is as follows; Ovary is 8.2${\pm}$3.1 g in left and 12.5${\pm}$5.6 g in right. Oviduct is 1.7${\pm}$0.6 g in left and 1.9${\pm}$0.9 g in right. Uterine horn is 199${\pm}$14.2 g in left and 221${\pm}$111.2g in right. Uterine body and cervix are 58.2${\pm}$46.5 and 126.7${\pm}$103.3 g.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• v.5 no.1
• /
• pp.1-10
• /
• 2002

Purpose; Dysregulation of gastric epithelial cell proliferation and apoptosis are important in development of ulcer, atrophy and neoplasia in Helicobacter pylori (H. pylori) infection. The aim of this study was to investigate the effect of infection of H. pylori on gastric epithelial cell proliferation and apoptosis in children. Methods: Histological grading by updated Sydney system, PCNA immunostaining and TUNEL method were performed in H. pylori positive (N=58) and negative (N=40) gastric biopsy specimens. Results: In H. pylori positive children, there were significantly higher grade of polymorphonuclear neutrophil activity (P=0.000), chronic inflammation (P=0.000), epithelial damage (P=0.000) and lymphoid follicles (P=0.000) than in H. pylori negative children. Intestinal metaplasia was not seen in H. pylori positive children. PCNA index was significantly different between H. pylori positive children ($67.8{\pm}18.13$) and H. pylori negative children ($54.8{\pm}14.46$, P=0.000). There was positive correlation between PCNA index and H. pylori density (r=0.277, P=0.007), polymorphonuclear neutrophil activity (r=0.280, P=0.007) and chronic inflammation (r=0.284, P=0.006). Apoptosis index of H. pylori positive children ($0.44{\pm}0.447$) was significantly higher than of H. pylori negative children ($0.14{\pm}0.196$, P=0.000). There was positive correlation between apoptosis index and H. pylori density (r=0.472, P=0.000), polymorphonuclear neutrophil activity (r=0.370, P=0.001) and chronic inflammation (r=0.483, P=0.000). There was positive correlation between PCNA index and apoptosis index (r=0.353, P=0.003). Conclusion: The PCNA and apoptosis index in H. pylori positive children were significantly higher than in H. pylori negative children. This study suggested that gastric epithelial cell proliferation and apoptosis are important to pathogenesis of H. pylori infection in children.

• Clinical and Experimental Reproductive Medicine
• /
• v.34 no.1
• /
• pp.49-56
• /
• 2007

Objective: To evaluate the effect of short coasting, by withdrawing both gonadotropins and GnRH agonist (GnRHa), on the prevention in severe ovarian hyperstimulation syndrome(OHSS) without compromising pregnancy outcome. Method: Thirty-seven women who had been coasted during COH for IVF were coasted when $\geq$20 follicles > 15 mm with serum E$_2$ level of 4,000 pg/ml were detected. Coasting was initiated for one or two days depending on the status of follicle on ultrasound and serum E$_2$ level. Both gonadotropin and GnRHa were withheld for coasting. Retrospective study was carried and changes of serum E$_2$ levels, number of oocytes retrieved, fertilization rate, pregnancy rate were compared and analyzed. Results: The mean serum E$_2$ level fell from 6,993 pg/ml on the onset of coasting to 3,396 pg/ml on the day of hCG administration. The mean number of oocytes retrieved and fertilization rate were 15.7 and 70.0%, respectively. Fifteen patients were pregnant (40.6%) and implantation rate was 15.2%. Twenty-six (70.3%) patients were coasted for one day and 11 (29.7%) were coasted for two days. The mean decrease rate of serum E$_2$ level was 43% in one day coasting group and 15% (1$^{st}$ day) and 81% (2$^{nd}$ day) in two day coasting group. The pregnancy outcome was similar between the two groups. After coasting, no severe or moderate OHSS occurred in any patients and mild OHSS occurred in 3(8.1%) patients. Conclusions: Coasting for one or two days can be used successfully in the prevention of OHSS without compromising IVF cycle outcome.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.47 no.3
• /
• pp.255-263
• /
• 2021

Soybean extract is known to play an important role in preventing and treating diseases associated with aging, cancer, obesity, and cardiovascular disease. A recent has revealed that soybean extract has a potent effect on hair growth in in vitro, in vivo, and clinical studies. Recently, it has been reported that their fermented extracts exhibit numerous and high efficacy, as compared to general extracts. However, the underlying mechanisms that induce hair growth after using fermented soybean extract are not well understood. The present study aimed to determine the effects of fermented Jeju soybean (FJS) extract on hair growth, with a focus on the underlying mechanisms similar to those of minoxidil. We conducted in vitro and ex vivo investigations and clinical studies. FJS extract enhanced dermal papilla cell proliferation, VEGF levels, and potassium channel opening. Moreover, it promoted human hair follicle elongation. These effects were comprehensively demonstrated in the clinical results, in which FJS extract-containing shampoo improved hair density after 24 weeks of utilization. Collectively, the results of this study demonstrate that FJS extract promotes hair growth and inhibits hair loss through a mechanism similar to that of minoxidil in hair follicles.

• Korean Journal of Food Science and Technology
• /
• v.54 no.1
• /
• pp.43-51
• /
• 2022

Regulation of the hair follicle cycle in association with dermal papilla cells is one of the most interesting targets for promoting hair regrowth. In this study, we examined whether steam-dried Betula platyphylla extracts (BPE) promote hair growth by upregulating in vitro and in vivo responses of dermal papilla cells. The data showed that BPE3 contained high amounts of phenolic compounds with higher antioxidant effects and increased hair growth-related genes, including fibroblast growth factor7 and Wnt7b, in dermal papilla cells. Notably, BPE3 effectively enhanced the formation of hair follicles by increasing FGF7, Wnt7b, and vascular endothelial growth factor in C57BL/6N dorsal skins. Additionally, BPE3 significantly decreased the expression of inflammatory repertoires, inducible nitric oxide synthase, interleukin-6, and cyclooxygenase 2. Several small molecules, such as betulin and unsaturated fatty acids, support the pharmacological activity of BPE3. In conclusion, BPE3 effectively promoted hair growth by activating dermal papilla cells and enhancing hair follicle cycles by attenuating the inflammatory environment in the scalp.