• Korean Journal of Microbiology
• /
• v.43 no.3
• /
• pp.151-158
• /
• 2007

The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with $1{\times}10^9$ PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.

• Korean Journal of Animal Reproduction
• /
• v.8 no.1
• /
• pp.36-45
• /
• 1984

This study was carried out to investigate the changes in ovary in repeatedly superovulated rabbits. A total of 57 New Zealand White and Californian, 25 mature virgin and 32 immature does were used in this study. For induction of repeated superovulation, PMSG and HCG were injected at 17-day and 30-day intervals for mature does and 17-day intervals for immature ones. The repeatedly superovulated does at 17-day intervals were induced luteolysis of pseudopregnant corpus luteum with PGF2${\alpha}$ on Day 8 to 9 p.c. The effect of repeated superovulation on reproductive organs was investigated on Day 3 p.c. in mature does and on Day 3 and 6 p.c. in immature ones, respectively. 1. In mature virgin does, the number of ovulation points in the 2nd and 3rd superovulation period averaged 7.0 and 5.0 at 17-day intervals and 13.4 and 6.0 at 30-day intervals, respectively. These numbers were statistically similar to 9.5 ovulation points in the control. However, there were less (p<0.05) ovulation points in those periods compared with 22.1 ovulation points in the 1st superovulation period. 2. In immature does, the number of ovulation points in the 2nd and 3rd superovulation period averaged 5.3 and 2.3, respectively. These numbers were significantly (p<0.05) decreased than 17.1 ovulation points in the 1st periods. The number of ovulation points in the 2nd superovulation period was similar to that in the control, but there was a significant (p<0.05) decrease in the number of ovulation points in the 3rd period as compared to the control. 3. In mature virgin does, the number of visible normal and hemorrhagic follicles (>1.0mm diameter) on day 3 p.c. averaged 19.1 and 8.9 in the 1st superovulation period, respectively. In the 2nd 3rd superovulation period, the number of normal follicles averaged 8.3 and 15.5 at 17-day intervals and 17.8 and 14.5 at 30-day intervals. The number of hemorrhagic follicles in the 2nd and 3rd superovulation period averaged 6.3 and 2.0 at 17-day intervals and 5.2 and 7.8 at 30-day intervals, respectively. There was a slight decrease, although not significant, in the number of normal and hemorrhagic follicles in the 2nd and 3rd period at 17-day intervals compared to that in the 1st period. 4. In immature does, the number of visible normal follicles on day 3 and day 6 p.c. in the 1st superovulation period averaged 27:3 and 26.1, respectively. The follicles on day 3 p.c. tended to increase slightly more than that in the cortrol, but the average number of normal follicles on day 6 p.c. did not differ from that in the control. The number of visible hemorrhagic follicles on day 3 and day 6 p.c. in the 1st of follicles in the 1st superovulation period average 10.2 and 9.9, respectively. There was a slight increase in the number of follicles in the 1st period compared to that in the control. In the 2nd and 3rd superovulation period, the number of normal follicles revealed a slight decrease in the 3rd period, but the number of hemorrhagic follicles was not different between periods. 5. The number of growing follicles with incipient intral formation on day 3 p.c. in mature does of the 1st superovulaton period average 29.7 and the average number of growing follicles in the 3rd period was 26.7 at 17-day intervals and 31.0 at 30-day intervals, respectively. These numbers did not differ from that in the control. In immature does, the number of growing follicles averaged 57.7, 45.0 and 59.3 in the 1st, 2nd and 3rd superovulation period, respectively. There was a slight but not significant decrease in the number of growing follicles in the 3rd period compared to that in the control.

• Korean Journal of Animal Reproduction
• /
• v.21 no.3
• /
• pp.293-302
• /
• 1997

Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

• Korean Journal of Animal Reproduction
• /
• v.21 no.3
• /
• pp.255-264
• /
• 1997

The purpose of this study was attempted to investigate the number changes of the growing and mature follicles in ovary following repeats of pregnant mare serum gonadotropin(PMSG) treatments for superovulation in nulliparous rats. Thirty two rats(Sprague-Dawely, about 200~250 gm) were randomized into 4 groups. Control group rats were sacrified at estrus phase confirmed by vaginal smear. PMSG-treated group 1 rats, PMSG-treated group 2 rats and PMSG-treated group 3 rats were sacrified at 48 hrs after injection once with PMSG 25 IU, after 2 repeated injection by a week interval, and 3 repeated injection, respectively. The uteri and ovaries of rats were removed and weighed and then were observed grossly and serial sections of all ovaries and some sections of uteri by paraffin embedding were stained with H-E. Number of ovarian follicles about 3 grades of small, middle and large follicles from seondary and follicles were investigated by LM photographies of ovary preparations. The criteria of the small, middle, and large follicles were based as small follicle with preantral follicles with 2~4 layers of granulosa cells surrounding the oocyte, as secondary follicles with more than 5 layers of granulosa cells and early signs of antral cavity or with small clefts on either side of the oocytes, and as tirtiary follicles with a single medium sized antral cavity or large well-formed antral cavity, respectively. In gross findings, the wall of the uteri in control group were thin, and those in 3 PMS-treated group were markedly thickened and some uterine lumen of those filled with fluid. In histological findings, the walls of the uteri from 3 PMSG-treated groups were hypertrophied and their blood and lymph vessels were dilated than those of control group. The ovaries fo 3 PMSG-treated groups were more increased in size and the cortexes were more developed and increased in width but there are no difference of development and changes in 3 PMSG-treated groups. The weight of the uteri and ovaries per rat in PMSG -treated group 1, 2 and 3 were a, pp.ared to be significantly increased 171.4$\pm$47.6%, 162.3$\pm$43.9%, 206.9$\pm$30.4%, respectively than those of control groups. The mean number of follicle per ovary in control group were a, pp.ared to be 17.1$\pm$3.5, 46.2$\pm$14.5, and 74.3$\pm$22.7 at large, middle and small follicles, respectively and total number of these 3 grade follicles per ovary were a, pp.ared to be 137.7$\pm$31.7. The mean number of follicle per ovary in PMSG-treated group 1 were a, pp.ared to be 25.6$\pm$7.3, 78.1$\pm$29.9, and 83.2$\pm$34.0, at large, middle and small follicles, respectively and total number of these 3 grade follicles were a, pp.ared to be 187.5$\pm$58.8. The mean number of follicle per ovary in PMS-treated group 2 were a, pp.ared to be 21.9$\pm$5.2, 67.8$\pm$16.8, and 68.0$\pm$14.9 at large, middle and small follicles, respectively and total number of these 3 grade follicles were a, pp.ared to be 157.7$\pm$26.2. The mean number of follicle per ovary in PMS-treated group 3 were a, pp.ared to be 21.7$\pm$4.8, 61.5$\pm$17.0, and 59.7$\pm$16.2 at large, middle and small follicles, respectively and total number of these 3 grade follicles were a, pp.ared to be 143.5$\pm$29.6. The number of follicles in PMSG-treated group 1 a, pp.ared to be more number than other 2 PMSG-treated gruops and tended to be decreased by frequency of PMSG-treatment.

• Applied Microscopy
• /
• v.41 no.2
• /
• pp.109-116
• /
• 2011

Polycystic ovary syndrome (PCOS) is hormonal imbalance condition as the endocrine and metabolic disorder that induces the infertility and various complications in reproductive age women. Estradiol valerate (EV) is used hormone replacement therapy in menopausal women and is reported that excessive administration of EV induces the PCOS. Nerve growth factor (NGF) is the factor to regulate the survival and maturation of developing neuronal cell and is also synthesized in ovary. And NGF is overexpressed in EV-induced polycystic ovary (PCO) as previously reported. Therefore, this study examined the possibility of NGF as can be used the biological marker in diagnosis of PCOS, the hormonal imbalance condition, using PCO and CHO (chinese hamster ovarian) cell lines. The concentration of EV treatment is optimized a 1 mg as not influence on the proliferation of CHO cell but 2 mg and 3 mg of EV treatment have the inhibition effect at initial stage. The morphological change was not observed in CHO cell after dose dependent manner treatment of EV. Expression of NGF mRNA and protein is significantly increased at 30 min after EV treatment in CHO cells compared to that of control. And NGF protein expression is strongly increased in PCO tissue, which observed many follicular cysts compared to normal ovary tissue. Taken together, overexpression of NGF may be act as a molecule to induce an abnormal development of follicle, suggesting that NGF can be used as a biological marker in diagnosis of PCOS.

• Journal of Embryo Transfer
• /
• v.21 no.3
• /
• pp.217-223
• /
• 2006

This study was carried out principally to obtain the basic data for the improvement of the reproductive performance and production using plasma progesterone assay and ultrasonography in dairy cow. The results obtained from this studies were as follows. The results of reproductive examination in 85,983 cows were ovarian diseases 40,399 (47.0%), uterine diseases 11,912 (13.9%), pregnancy or pregnant failures 26,587 (30.9%), adhesion of reproductive tracts 172 (0.2%), freemartin 8 (0.01%), and others 6,905 (8.3%), respectively. The treatment status of reproductive dysfunction in 30,241 cows were silent heat or error of estrus detection 14,909 (49.3%), follicular cysts 3,750 (12.4%), luteal cysts 907 (3.0%), inactive ovaries 665 (2.2%), granulosa cell tumor of ovary 3 (0.01%) and endometritis 6,986 (23.1%), respectively. The indices of reproductive efficiency after the periodical examination of reproductive status were as follows; the mean intercalving inteual was reduced from 475 days at the first examination to 381 days at the last examination of reproductive status, the mean interval calving to conception was reduced from 186 to 98 days, the mean interval calving to first service was reduced from 106 to 66 days, the cows showing heat by 60 days postpartum were increased from 32 to 90%, the mean conception rate to first service was increased from 42 to 64%, and the mean service per conception was reduced from 2.6 to 1.8 times, respectively.

• Journal of Life Science
• /
• v.27 no.11
• /
• pp.1245-1255
• /
• 2017

Most cancer cells produce ATP predominantly through glycolysis instead of through mitochondrial oxidative phosphorylation, even in the presence of oxygen. The phenomenon is termed the Warburg effect, or the glycolytic switch, and it is thought to increase the availability of biosynthetic precursors for cell proliferation. EMTs have critical roles in the initiation of the invasion and metastasis of cancer cells. The glycolytic switch and EMT are important for tumor development and progression; however, their correlation with tumor progression is largely unknown. The Snail transcription factor is a major factor involved in EMT. The Snail expression is regulated by distal-less homeobox 2 (Dlx-2), a homeodomain transcription factor that is involved in embryonic and tumor development. The Dlx-2/Snail cascade is involved in Wnt-induced EMTs and the glycolytic switch. This study showed that in response to Wnt signaling, the Dlx-2/Snail cascade induces the expression of PFKFB2, which is a glycolytic enzyme that synthesizes and degrades fructose 2, 6-bisphosphate (F2,6BP). It also showed that PFKFB2 shRNA prevents Wnt-induced EMTs in the breast-tumor cell line MCF-7. The prevention indicated that glycolysis is linked to Wnt-induced EMT. Additionally, this study showed PFKFB2 shRNA suppresses in vivo tumor metastasis and growth. Finally, it showed the PFKFB2 expression is higher in breast, colon and ovarian cancer tissues than in matched normal tissues regardless of the cancers' stages. The results demonstrated that PFKFB2 is an important regulator of EMTs and metastases induced by the Wnt, Dlx-2 and Snail factors.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.33 no.5
• /
• pp.431-438
• /
• 2000

To clarify the annual reproductive cycle in a rockfish, Sebastes schlegeli, monthly changes in gonadosomatic index (GSI), hepatosomatic index (HSI) and histological feature of gonads and plasma levels of sex steroid hormones ($estradiol-l7{\beta},\;17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one,\;testosterone\;and\;11-ketotestosterone$) were investigated. The annual reproductive cycle in females could be divided into 5 periods as follows: 1) recovery period (June to September): serum level of $estradiol-l7{\beta}$ increased gradually; 2) vitellogenesis period (Septemer to february) : vitellogenic oocytes were obsewed, GSI sustained high value, and serum level of $estradiol-l7{\beta}$ increased; 3) gestation period (February-April): developing larva showed in the ovary, and serum levels of $17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one$ and testosterone increased; 4) partrition period (April to May) : larva were delivered, and value of GSI and serum levels of hormones decreased rapidly; 5) resting period (May to June) : value of GSI and serum levels of $estradiol-l7{\beta}$ and testosterone remained low. The annual reproductive cycle in males could be divided into 6 periods; 1) early maturation period (April to June): value of GSI and serum levels of hormones incresed gradually, cyst of spermatogonia incresed in number, and a small number of cyst of spermatocyte was observed; 2) mid-maturation perid (June to September); value of GSI and serum levels of hormones increased, and germ cells in many cysts were undergoing active sperrnatogenesis; 3) late maturation period (September to November) : value of GSI and serum levels of hormones remained high and spermatozoa were released into the lumina of the seminal lobules; 3) spermatozoa dischaging period (Nobember to December) : the lumina of the seminal lobules were enlarged and filled with mature spermatozoa; 4) degeneration period (December to Februauy)i value of GSI decresed and cyst of spermatocyte were decresed in number; 5) resting period (December to April) : no histological changes of testes were observed, and value of GSI and serum levels of hormones remained low. In November, the lumina of the seminal lobules were filled with mature spermatozoa and sperm masses were present in the ovarian cavity. Thus, copulation in this species occurred in November and December.

• Korean Journal of Veterinary Research
• /
• v.37 no.1
• /
• pp.71-78
• /
• 1997

This study was designed to investigate the number of the growing and mature follicles following gonadotrophin treatments for superovulation in mature rats. Eighteen mature rats (Sprague-Duwely, initially 190~230gm) were randomly alloted into 3 groups. One group was control group, another FSH-treated group was injected intramuscularly with 0.5 units of follicular stimulating hormone (FSH) / rat, and third PMS and HCG-treated group was intramuscularly injected with 20~25IU of pregnant mare serum (PMS) / rat and then at the 48 hrs later, with 20~25IU of human chorionic gonadotrophin (HCG) / rat. The uteri and ovaries of rats were collected and then were observed grossly and serial sections of paraffin embedding ovaries were stained with H-E. Number of ovarian follicles by following 3 grades of large, middle and small follicles from secondary and tertiary follicles were investigated by LM photography of preparations. Small follicles were classified as secondary follicles of preantral follicles with more than 2 layers of granulosa cells surrounding the oocyte and middle follicles were classified as secondary follicles with early signs of antral cavity or with more than one small cavity on either side of the oocytes and large follicles were classified as tertiary follicles with a single medium sized antral cavity or large well-formed antral cavity. In gross findings, the uteri were slightly swelling in FSH-treated group and markedly swelling or filled with fluid in the uterine lumen in PMS and HCG-treated group. In histological findings, the shape and size of the follicles were diverse in middle and large follicles of FSH-treated group and PMS and HCG-treated group, and proportion of atretic follicles was increased in FSH-treated group and PMS and HCG-treated group than those in control group. The uteri of FSH-treated group and PMS and HCG-treated group were hypertropied or filled with fluid in the lumens and walls of uteri. The wall tissue layers were flattened and their blood and lymph vessels were dilated. The mean number of follicle per ovary in control group were appeared to be $17.1{\pm}5.6$($14.0%{\pm}4.6%$), $37.8{\pm}9.1$($30.9{\pm}7.4%$) and $67.6{\pm}30.1$($55.2{\pm}24.6%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $122.5{\pm}40.0$. The mean number of follicle per ovary in FSH-treated group were appeared to be $22.8{\pm}7.0$($17.4%{\pm}5.3%$), $43.4{\pm}6.6$($33.2{\pm}5.1%$) and $64.5{\pm}13.0$($49.3{\pm}9.9%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $130.7{\pm}16.6$. The mean number of follicle per ovary in PMS and HCG-treated group were appeared to be $29.7{\pm}11.0$($16.3%{\pm}6.0%$), $61.9{\pm}17.2$($33.9{\pm}9.4%$) and $91.1{\pm}28.2$($49.9{\pm}15.4%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $182.6{\pm}32.7$. The above findings reveal that large follicles were increased 29.8% in FSH-treated group and 73.7% in PMS and HCG-treated group than those in control group and in histologic findings, proportion of atretic follicles were more increased in ovaries with more number of more developing follicles.

• Radiation Oncology Journal
• /
• v.16 no.3
• /
• pp.225-231
• /
• 1998

Purpose : To determine whether TNF-$\alpha$ increases the antitumor effect of radiotherapy in murine syngeneic tumor system. Materials and Methods : Syngeneic murine tumors of MCa-K or MCa-4 (mammary carcinoma), OCa-I (ovarian carcinoma), or HCa-I(hepatocarcinoma were grown in hind legs of C3Hf/HeJ mice. When tumors were grown to 6 mm in mean diameter mice were treated with TNF-$\alpha$, radiation, or combination of the both. Gamma-radiation was given as a single dose of 30 Gy for HCa-I and 15 Gy for other tumors using Cobalt-60 teletherapy unit. A novel TNF-$\alpha$ mutein developed in Korea, was intraperitoneally administered daily at a dose of 10 ug per mouse for 7 days. In combination of radiation and TNF-$\alpha$, the drug was started 1 hour after radiation. Tumor growth delay assay was used to measure the tumor response to the treatment. Results : Among 4 tested tumors, TNF-$\alpha$ alone showed significant antitumor activity in MCa-K and OCa-I tumors, which showed absolute growth delay (AGD) of 5.0 days and 6.5 days, respectively. In combination with radiation, TNF-$\alpha$ showed significant delay of AGD (41.1 days) in OCA-I compared to AGDs of TNF-$\alpha$ alone and radiation, i.e., 6.5 days and 26.9 days, respectively(p<0.05). Enhancement factor was 1.29 in OCa-I, which showed supraadditive effect. TNF-$\alpha$ did not show significant delay of AGDs in the remaining 3 tumors compared to AGDs of TNF-$\alpha$ alone and radiation. Conclusions: TNF-$\alpha$ alone showed antitumor effects in MCa-K and OCa-I. In combination with radiation, TNF-$\alpha$ acted in supraadditive way in OCa-I only. The results of this study imply that the combination of TNF-$\alpha$ and radiation has different therapeutic potential depending on tumor model and further study is advocated.