• Asian Pacific Journal of Cancer Prevention
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• v.16 no.18
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• pp.8503-8512
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• 2016

The purpose of this study was to provide a detailed explanation of the mechanism of bisanthracycline, WP 631 in comparison to doxorubicin (DOX), a first generation anthracycline, currently the most widely used pharmaceutical in clinical oncology. Experiments were performed in SKOV-3 ovarian cancer cells which are otherwise resistant to standard drugs such as cis-platinum and adriamycin. As attention was focused on the ability of WP 631 to induce apoptosis, this was examined using a double staining method with Annexin V and propidium iodide probes, with measurement of the level of intracellular calcium ions and cytosolic cytochrome c. The western blotting technique was performed to confirm PARP cleavage. We also investigated the involvement of caspase activation and DNA degradation (comet assay and immunocytochemical detection of phosphorylated H2AX histones) in the development of apoptotic events. WP 631 demonstrated significantly higher effectiveness as a pro-apoptotic drug than DOX. This was evident in the higher levels of markers of apoptosis, such as the externalization of phosphatidylserine and the elevated level of cytochrome c. An extension of incubation time led to an increase in intracellular calcium levels after treatment with DOX. Lower changes in the calcium content were associated with the influence of WP 631. DOX led to the activation of all tested caspases, 8, 9 and 3, whereas WP 631 only induced an increase in caspase 8 activity after 24h of treatment and consequently led to the cleavage of PARP. The lack of active caspase 3 had no outcome on the single and double-stranded DNA breaks. The obtained results show that WP 631 was considerably more genotoxic towards the investigated cell line than DOX. This effect was especially visible after longer times of incubation. The above detailed studies indicate that WP 631 generates early apoptosis and cell death independent of caspase-3, detected at relatively late time points. The observed differences in the mechanisms of the action of WP631 and DOX suggest that this bisanthracycline can be an effective alternative in ovarian cancer treatment.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.6
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• pp.806-811
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• 2002

A 90 d study was designed to investigate the effects of rumen undegradable protein (RUP) and a mixture of Cu, Zn and Mn proteinate (CZMP) on milk yield and composition and ovarian functions during rainy months. Twenty four Holstein${\times}$ indigenous cows in their 2nd and 3rd lactation were randomly allocated to total mixed rations (TMR) containing soy bean meal (SBM) as a source of rumen degradable protein (RDP), SBM plus CZMP, and formalin treated SBM (FSBM) as a source of RUP. Maximum and minimum temperature humidity index during the experimental period were 83.6-84.7 and 75.4-76.1. There were no differences (p>0.05) in intakes of dry matter, crude protein and net energy and in contents of butterfat, lactose and minerals. Cows on TMR containing FSBM not only lost less weight (-278, -467 and -433 g/d) with more intake of RUP (0.92, 0.58 and 0.59 kg/d) but also produced more milk (19.27, 18.23 and 18.13 kg/d) and 4% fat corrected milk (18.57, 17.57 and 17.51 kg/d) with more protein (3.06, 2.81 and 2.80%), solids-not-fat (8.69, 8.38 and 8.38%) and less milk urea N (9.3, 15.4 and 15.0 mg/dl) compared with those on TMR containing SBM and SBM+CZMP, respectively (p<0.01). However, cows on TMR containing SBM and SBM+CZMP did not differ in these respects (p>0.05). Whereas incidence of cystic ovaries at 20 and 90 d pospartum was less (p<0.01) in cows on TMR containing SBM+CZMP (37.3 and 12.5%) than those on TMR containing SBM (62.5 and 25%), it was nil for cows on TMR containing FSBM. Cows in all three group differed (p<0.01) from each other for the recurrence of first observed estrus with those on TMR containing FSBM having least days (22, 36 and 47 d) compared with their counterpart on TMR containing SBM+CZMP and SBM, repectively. The results suggest that RUP is one of the limiting factors affecting milk yield and its composition and ovarian functions during early lactation of dairy cows in the tropics.

• Development and Reproduction
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• v.13 no.4
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• pp.341-351
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• 2009

The effects of serotonin (5-hydroxytryptamine, 5-HT), ecdysone (20-hydroxyecdysone, 20-E) and HCG (human chorionic gonadotropin) on the ovarian maturation of the fleshy shrimp, Fenneropenaeus chinensis were investigated. Adult female fleshy shrimps were injected with 5-HT ($5{\mu}g$/g body weight (BW) or $20{\mu}g$/g BW), 20-E ($5{\mu}g$/g BW, $10{\mu}g$/g BW or $20{\mu}g$/g BW) or HCG (5 IU/g BW or 10 IU/g BW) three times at 5-day intervals and were sacrificed 10 days after the last injection. The effects were compared against an intact control group and sham control group which received the injection of sterile saline solution as a vehicle. 20-E and HCG showed no effectiveness on ovarian maturation in fleshy shrimps. However flesh shrimp given 5-HT of $20{\mu}g$/g BW showed significant increase in gonadosomatic index (GSI) in early spring. The results suggest that treatment of 5-HT of $20{\mu}g$/g BW or more is useful approach for the nursery production of the fleshy shrimp, Fenneropenaeus chinensis aquaculture.

• Journal of Veterinary Clinics
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• v.15 no.2
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• pp.307-318
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• 1998

Accuracy of rectal palpation and ultrasonography for differential diagnosis of subestrous dairy trows were investigatedl using the result of pIRsma progesterone assay. The ovaries were examined 2 times of 10 days interval in 520 posearom and postinsemination subestroHs dairy cows, using rectal palpation and B-mode transrectal ultrasonography. The results of rectal palpation, ultrasonographic examination and measurement of plasma progesterone profiles in 520 subestrous dairy cows were silent brat or error of estrus detection 303 (58.3%), persistent corpus luteum 59 (11.3%), follicular cyst 37 (7.1%), luteal cyst 16 (3.1%), inactive ovary 9 (1.7%), granulosa tumor 1 (0.2%), hydmsalphinx 1 (0.2%), endomehris 81 (15.6%), pyometra 12 (2.3%) and mummified fetus 1 (0.2%), respectively. Accuncy of rectal palpation and ultrasonography for diagiosing ovarian disordeir based on plasma progesterone profiles were silent heat or error of estrus detection 80.5% and 96.7%$\boxUl$ persistent corpus luteum 57.6% and 94.9%, follicular cyst 62.5% and 91.9%1 luteal cyst 62.5% and 87.5%, maclive ovary 55.6% and 88.9% and granulosa cell tumor 100% and 100%, respectively. Acnuucy of rectal palpation for diagnosing uterine disorders based on ultrasonography was pyometra 75.0%1 endometritis 51.9% and mummified fetus 100%, respectively. Cbaracteristic ultrasonographic appearances of ovaries in subestrous dairy cows were as follows; Silent heat or error of estrus detection: anechoic follicle or hypoechoic corpus luteum than ovarian stroma was alternately present on Day 0 (first examination) and Day 10. Follicular cyst: uniformly nonechogenic ovarian structure $\geq $ 25 mm in diameter with a wall < 3 mm was present in ipsilateral on Day 0 and Day 10. Luteal cyst: luteal cyst was similar to follicular cysts but thickness of cystic wall was $\geq $ 3 mm. Inactive ovary : structures within ovaries was not present on Day 0 Bnd Day 10. Characteristic uthssonograpsc appearances of uterus in subestrous dairy cows were as follows; Endometritis: characterized by uterine lumen containing fluid in which 'snowy'echogenic particles art suspended. Pyometra: ultrasonographic appearance of pyometra was diffuse echogenic particles distributed in fluid within the distended uterus, and a thickened uterine wall. These results indicated that ultrasonography was practical far diagnosing reproductive disorders. To diagnosing ovarian disorders, ultrasonography should be carried out 2 times of 10 days interval and rndometritis should be differentiated with uterus of luteal phase in normal cycling cows.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.1
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• pp.8-16
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• 1998

Sexual maturation and reproductive cycle of the goldeye rockfish, Sebastes thompsoni were investigated under photomicroscopy. Samples were collected monthly in the coastal water of Samcheonpo ($34^{\circ}55'N$ ), Korea from November 1995 to October 1996, The ovary consists of several ovarian lamellae originated from ovarian outer membrane. Oogonia which are originated from the inner surface of the ovarian lamella protrude to the ovarian cavity in oocyte stage, and they ave suspended by the egg stalk. The testis is seminiferous tubule type in internal structure. Seminiferous tubule consists of many testicular cysts which contain numerous germ cells in same developmental stage. Biological minimum size of female and male were 19.5 cm and 21.5 cm in total length, respectively. Gonadosomatic index (GSI) of female was the highest (9.56) in March and the lowest (0.15) in August. GSI of male was the highest (0.25) in February and the lowest (0.04) in July. Reproductive cycle was classified into the following successive stages: in female, growing (October and November), maturation ( $December\~February$), gestation (March), parturition and recovery ($April\~June$) and resting ($July\~September$), and in male, growing ($September\~November$), maturation ( December and January), ripe and copulation ( February and March) and degeneration and resting ($April\~August$).

• Korean Journal of Animal Reproduction
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• v.13 no.3
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• pp.127-133
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• 1989

This study was undertaken to understand development stage of ovary and changes of hormones concentration from February to May in the Israeli carp, Cyprinus carpio. The results obtained in these experiments are as follows ; 1. Serum LH levels began to increase sharphy in March, coinciding with the onset of rapid ovarian development. 2. LH levels were well correlated with changes in gonadosomatic index. 3. Dramatic increase in gonadosomatic index occurred during the months of March. 4. Ripe stage(Stage Ⅴ) rapidly increase in March. 5. Early perinucleolus oocyte radpily develop into late perinucleolus oocytes in March. 6. The vitellogenic phase begins as these late perinucleolus oocytes become transformed into early maturing oocytes through the accumulation of yolk. 7. The cytoplasm completely fills with yolk as oocytes reach the late maturing stage. 8. Changes in the microscopic appearances of the ovaries were well correlated with changes in both gonadosomatic index and macroscopic appearance. 9. It is concluded from these observations that LH plays a major role in sexual maturation of the Israeli carp.

• Animal cells and systems
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• v.5 no.3
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• pp.225-229
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• 2001

Nek2 is a mammalian protein kinase that is structurally homologous to NIMA, a mitotic regulator in Aspergillus nidulans. To understand cellular processes in which Nek2 participates during mammalian development, we investigated the expression and subcellular localization of Nek2 in vivo. The Nek2 protein was detected in spermatocytes and in a fraction of actively dividing ovarian follicle cells and of embryonic tissues. We also observed that Nek2 was localized in both the nucleus and centrosome in embryonic cells. Such localization pattern supports the proposal that Nek2 is a mitotic regulator that is involved in multiple cell cycle events during mammalian development.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.277-282
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• 1996

Serum inhibin concentrations, determimed by radioimmunoassay, were measured in women undergoing pituitary suppression with Decapeptyl and subsequently ovarian stimulation with Highly Purified-Metrodin(HP-FSH) to appraise follicular development. Early follicular basal serum inhibin level correlated with the number of oocytes retrieved(r=0.89, n=8, p<0.05). The number of oocytes retrieved showed a significant correlation with serum inhibin level on the day of hCG administration(r=0.73, n=8, p<0.05). The number of mature oocytes showed a significant correlation with serum inhibin level on the day of hCG administration(r=0.73, n=8, p<0.05). These data suggest that: (1) In the early follicular phase, basal serum inhibin may be a valid index to predict ensuing follicular growth : (2) In the preovulatory phase, maximum serum inhibin may be one of the indexes of follicular development during hyperstimulation cycles.

• Clinical and Experimental Reproductive Medicine
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• v.22 no.2
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• pp.155-161
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• 1995

The present investigation has been undertaken to elucidate the differentiation mechanism the uterus which is the environment of the embryo development, by demonstrating the role of ovarian steroids hormone in the decidualization of the pseudopregnant rat uterus. To determine the effect of ovarine steroids and artificial stimulation (trauma) on the differenciation of the uterine endometrium and decidualization for implantation, attempt was made to measure concentrations of serum estradiol($E_2$), progesterone($P_4$) and nuclear $P_4$ receptor in the traumatized and non-traumatized uterine tissue of the pseudopregnant rat. The results obtained are as followings : The concentration of serum $E_2$ on day 9(implantation stage) was similar in both of intact pseudopregnant rat(47.63pg/ml) and normal pregnant rat(40.71pg/ml). And among the treated groups, $E_2$ concentration was highest in the $E_2$ treated group in comparision with intact control group(relative value; 73.27%). The concentration of serum $P_4$ was also highest in the $P_4$ treated group(23.12pg/ml). Relative value of $P_4$ treated group in comparision with intact group(24.88pg/ml) was 92.93%. The nuclear $P_4$ receptor levels in the artificial traumatized groups were higher compared with the non-traumatized control groups. This study, therefore, clearly demonstrates that the methods for inducing pseudopregnant (vagina tapping;120/min) and inducing decidualization(oil injection; 0.1ml/uterine horn) appear to be effective, $P_4$ appears to be effective in the differenciation of the uterine endometrial tissue for the implantation process. Concentration of serum $P_4$ seems to be well correlated with the level of the nuclear $P_4$ receptor during the early embryo development. These results seem to be well correlated with ALPase activities in the normal and pseudopregnant rat uterus shown in the previous study.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.1
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• pp.18-23
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• 2011

Objective: Peroxiredoxins (Prxs) play an important role in regulating cellular differentiation and proliferation in several types of mammalian cells. This report examined the expression of Prx isotype I in the rat ovary after hormone treatment. Methods: Immature rats were injected with 10 IU of pregnant mare's serum gonadotropin (PMSG) to induce the growth of multiple preovulatory follicles and 10 IU of human chorionic gonadotropin (hCG) to induce ovulation. Immature rats were also treated with diethylstilbestrol (DES), an estrogen analogue, to induce the growth of multiple immature follicles. Northern blot analysis was performed to detect gene expression. Cell-type specific localization of Prx I mRNA were detected by in situ hybridization analysis. Results: During follicle development, ovarian Prx I gene expression was detected in 3-day-old rats and had increased in 21-day-old rats. The levels of Prx I mRNA slightly declined one to two days following treatment with DES. A gradual increase in Prx I gene expression was observed in ovaries obtained from PMSG-treated immature rats. Furthermore, hCG treatment of PMSG-primed rats resulted in a gradual stimulation of Prx I mRNA levels by 24 hours (2.1-fold increase) following treatment, which remained high until 72 hours following treatment. In situ hybridization analysis revealed the expression of the Prx I gene in the granulosa cells of PMSG-primed ovaries and in the corpora lutea of ovaries stimulated with hCG for 72 hours. Conclusion: These results demonstrate the gonadotropin and granulosa cell-specific stimulation of Prx I gene expression, suggesting its role as a local regulator of follicle development.