• Journal of Animal Reproduction and Biotechnology
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• v.39 no.1
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• pp.19-30
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• 2024

Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

• Development and Reproduction
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• v.16 no.3
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• pp.205-217
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• 2012

The ovarian cycle, the biological minimum size, and artificial spawning frequency by artificial spawning induction of the female hard clam, Meretrix petechialis, were investigated by histological observations and morphometric data. The ovarian cycle of this species can be classified into five successive stages: early active stage, late active stage, ripe stage, partially spawned stage, and spent/inactive stage. The spawning period was from June to September, and the main spawning occurred between July and August when the seawater temperature exceeds over $20^{\circ}C$. The biological minimum size (shell length at 50% of first sexual maturity) in females were 40.39 mm in shell length (considered to be two years of age), and all clams over 50.1 mm in shell length sexually matured. In this study, the mean number of the spawned eggs by spawning induction increased with the increase of size (shell length) classes. In case of artificial spawning induction for the clams > 40.39 mm, the number of spawned eggs from the clams of a sized class was gradually decreased with the increase of the number of the spawning frequencies (the first, second, and third spawning). In the experiments of artificial spawning induction during the spawning season, the interval of each spawning of this species was estimated to be 15-18 days (approximately 17 days).

• Clinical and Experimental Reproductive Medicine
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• v.26 no.3
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• pp.355-362
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• 1999

Objective: This study was performed to compare the clinical response to controlled ovarian hyperstimulation (COH) of in vitro fertilization and embryo transfer (IVF-ET) according to the size of baseline ovarian cyst. Method: From February 1992 to March 1999, a retrospective analysis was done of 272 cases who underwent COH using mid-luteal phase long protocol of gonadotropin-releasing hormone agonist (GnRH-a) for IVF-ET. These cases were divided into four group; group 1 (n=63) had cysts with mean diameters between 20.0 and 29.0 mm on their baseline ultrasound on cycle day 3, group 2 (n=57, $30.0{\sim}49.0mm$), group 3 (n=68, >50.0 mm) and control group (n=84). Cases were excluded according to the following criteria; pure male factor infertility, the presence of only one ovary, high CA-125 level and previous endometriosis. Results: There were no statistically significant differences between cases with baseline ovarian cyst <50.0 mm in diameter and control group in any of the parameters. However, cases with baseline ovarian cyst>50.0 mm in mean diameter needed more amount of human menopausal gonadotropin (hMG), showed significantly lower estradiol ($E_2$) level, the number of follicle >15.0 mm on the day of human chorionic gonadotropin (hCG) administration, the number of oocytes retrieved, the number of mature oocytes, and pregnancy rate compared with control group. Conclusion: This study suggests that cases with baseline ovarian cyst <50.0 mm in diameter do not adversely impact on IVF-ET outcome. However, cases with baseline ovarian cyst >50.0 mm in diameter had adverse effects on various parameters. Therefore, to improve the outcome of IVF-ET in these cases, ovarian cyst aspiration prior to initiating COH may be required.

• Journal of Gastric Cancer
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• v.11 no.1
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• pp.31-37
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• 2011

Purpose: We performed this study to evaluate the clinical presentation as well as the proper surgical intervention for ovarian metastasis from gastric cancers and these tumors were identified during postoperative follow-up. This will help establish the optimal strategy for improving the survival of patients with this entity. Materials and Methods: 22 patients (3.2%) with ovarian metastasis were noted when performing a retrospective chart review of (693) females patients who had undergone a resection for gastric cancer between 1981 and 2008. The covariates used for the survival analysis were the patient age at the time of ovarian relapse, the size of the tumor, the initial TNM stage of the gastric cancer, the interval to metastasis and the presence of gross residual disease after treatment for Krukenberg tumor. The cumulative survival curves for the patient groups were calculated with the Kaplan-Meier method and they were compared by means of the Log-Rank test. Results: The average age of the patients was 48.6 years (range: 24 to 78 years) and the average survival time of the 22 patients was 18.8 months (the estimated 3-year survival rate was 15.8%) with a range of 2 to 59 months after the diagnosis of Krukenberg tumor. The survival rate for patients without gross residual disease was longer than that of the patients with gross residual disease (P=0.0003). In contrast, patient age, the size of ovarian tumor, the initial stage of gastric adenocarcinoma, the interval to metastasis and adjuvant chemotherapy were not prognostic indicators for survival after the development of ovarian metastasis. Conclusions: Early diagnosis and complete resection are the only possible hope to improve survival. As the 3-year survival rate after resection of Krukenberg tumor is 15.8%, it seems worthwhile to consider performing tumorectomy as the second cytoreduction.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.3
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• pp.313-316
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• 2004

Ultrasonographic studies were conducted on eight Murrah buffaloes daily from day 6 postpartum (pp) onwards till day 77 pp to monitor changes in the cervix, uterine horn and ovarian follicular growth and development. The mean size of horn and cervix on day six ($9.07{\pm}0.74$ and $8.58{\pm}0.00cm$) decreased significantly to $4.09{\pm}0.09$ and $3.56{\pm}0.08cm$ by day 27 pp, respectively. Follicles in 50% of the buffaloes ovulated within 24 to 54 days pp and the size of the largest follicle on different days increased to more than 5 mm. The remaining 50 percent of animals ovulated after 65 days postpartum. Large size follicles (>8.5 mm) appeared in six out of eight buffaloes between 10 to 30 days pp and five animals had ovulated during early postpartum period. Waves pattern of follicular growth was observed during early postpartum period. Ovulatory follicles growth rate was more than the anovulatory follicles and increase in size was more as compared to the subordinate follicle. Anovulatory follicles persisted for longer period. Mean size of large follicle was more from day 6 to 41 pp and again from 50 to 65 pp in cyclic animals. Second large follicle were large during early postpartum (18days), thereafter, its size was more in acyclic animals. Small follicles population was less in cyclic animals upto day 50 postpartum. Mean medium size follicle growth pattern did not differ in cyclic and acyclic groups. Large size follicle number was more in cyclic group (5/8) during 14 to 20 days postpartum. Presence of large follicles (>8.5 mm) showed initiation of ovarian activity.

• Development and Reproduction
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• v.16 no.3
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• pp.185-193
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• 2012

For the gonadal sex management of younger longtooth grouper (Epinephelus bruneus), this work investigated the timing and histological process of ovary differentiation and oocyte development of longtooth grouper larvae and juvenile. Specimens (from 1 to 365 DAH) were collected for gonadal histological study from June 2008 to August 2009. Rearing water temperature was ranged from 20 to $24^{\circ}C$. The primordial germ cells could be observed from 10 to 15 DAH, while undifferentiated gonad occurs from 20 to 50 DAH in longtooth grouper. The initial ovarian phase was 60 to 110 DAH with the formation of ovarian cavity and the increased in size of gonad. The ovarian phase started at 140 DAH with appearance of oogonia. The gonad at 365 DAH appeared to have full of oogonia and primary growth stage oocyte. Formation of ovarian cavity indicates that the ovarian differentiation beginning at 60 DAH in longtooth grouper. The gonads in longtooth grouper differentiated directly into ovaries in all fish examined.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.8
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• pp.3273-3277
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• 2015

Network scale up (NSU) is a novel approach to estimate parameters in hard to reach populations through asking people the number of individuals they know in their active social network. Although the method have been used in hidden populations, advantages of NSU indicate that exploration of applicability to disease like cancer might be feasible. The aim of this study was to assess the application of NSU to estimate the size of the population of breast, ovarian/cervical, prostate, and bladder cancers in the South-east of Iran. A total of 3,052 (99% response rate) Kermanian people were interviewed in 2012-2013. Based on NSU, participants were asked about if they know any people on their social network who suffered from breast, ovarian/cervical, prostate, and bladder cancers, if yes, they should enumerate them. A total of 1,650 persons living with four types of cancers (breast, ovary/cervix, prostate, and bladder) were identified by the respondents. Totally, the prevalence of people living with the four types of cancers was 228.4 per 100,000 Kermanian inhabitants. The most prevalent cancer was breast cancer, at 168.9 per 100,000, followed by prostate cancer with 116.9, ovarian/cervical cancer with 99.8, and bladder cancer with 36.3 per 100000 Kerman city population. NSU values provide a usable but not very precise way of estimating the size of subpopulations in the context of the four major cancers (breast, ovary/cervix, prostate, and bladder).

• Clinical and Experimental Reproductive Medicine
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• v.32 no.3
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• pp.269-277
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• 2005

Objectives: Previously, we sought to compile a list of genes expressed during early folliculogenesis by using cDNA microarray to investigate follicular gene expression and changes during primordialprimary follicle transition and development of secondary follicles (Yoon et al., 2005). Among those genes, a group of genes related to the cell size growth was characterized during the ovarian development in the present study. Methods: We determined ovarian expression pattern of six genes related to the cell size growth (cyr61, emp1, fhl1, socs2, wig1 and wisp1) and extended into CCN family (${\underline{c}}onnective$ tissue growth factor/${\underline{c}}ysteine$-rich 61/${\underline{n}}ephroblastoma$-overexpressed), ctgf, nov, wisp2, wisp3, including cyr61 and wisp1 genes. Expression of mRNA and protein according to the ovarian developmental stage was evaluated by in situ hybridization, and/or semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and immunohistochemistry, respectively. Results: Among 6 genes related to the cell size growth, cyr61 and wisp1 mRNA was detected only in oocytes in the postnatal day5 mouse ovaries. cyr61 mRNA expression was limited to the nucleolus of oocytes, while wisp1 was expressed in the cytoplasm and nucleolus of oocytes, except nucleus. cyr61 mRNA expression, however, was found in granulosa cells from secondary follicles. The rest 4 genes in the cell size growth group were detected in oocytes, granulosa and theca cells. Cyr61 and Wisp1 proteins were expressed in the oocyte cytoplasm from primordial follicle stage. Especially, Cyr61 protein was detected in pre-granulosa cells, Wisp1 protein was not. By using RT-PCR, we evaluated and decided that Cyr61 protein is produced by their own mRNA in pre-granulosa cells that was not detected by in situ hybridization. cyr61 and wisp1 genes are happen to be the CCN family members. The other members of CCN family were also studied, but their expression was detected in oocytes, granulose and theca cells. Conclusions: We firstly characterized the ovarian expression of genes related to the cell size growth and CCN family according to the early folliculogenesis. Cyr61 protein expression in the pre-granulosa cells is profound in meaning. Further functional analysis for cyr61 in early folliculogenesis is under investigation.

• Animal cells and systems
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• v.5 no.3
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• pp.217-224
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• 2001

The present study examines the expression and regulation of gonadotropin-releasing hormone (GnRH) and its receptor (GnRH-R) mRNA levels during mouse ovarian development. A fully processed, mature GnRH mRNA together with intron-containing primary transcripts was expressed in the immature mouse ovary as determined by Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR). The size of ovarian GnRH mRNA was similar to that of hypothalamus, but its amount was much lower than that in the hypothalamus. Quantitative RT-PCR procedure also revealed the expression of GnRH-R mRNA in the ovary, but the estimated amount was a thousand-fold lower than that in the pituitary gland. We also examined the regulation of ovarian GnRH and GnRH-R mRNA levels during the follicular development induced by pregnant mare's serum gonadotropin (PMSG) and/or human chorionic gonadotropin (hCG). Ovarian luteinizing hormone receptor (LH-R) mRNA was abruptly increased st 48 h after the PMSG administration and rapidly decreased to the basal level thereafter. Ovarian GnRH mRNA level was slightly decreased at 48 h after the PMSG administration, and then returned to the basal value. GnRH-R mRNA level began to increase at 24 h after the PMSG treatment, decreased below the uninduced basal level at 48 h, and gradually increased thereafter. HCG administration did not alter ovarian GnRH mRNA level, while it blocked the PMSG-induced increase in GnRH mRNA level. Taken together, the present study demonstrates that the expression of GnRH and GnRH-R mRNA are regulated by gonadotropin during follicular development, suggesting possible intragonadal paracrine roles of GnRH and GnRH-R in the mouse ovarian development.

• Journal of Yeungnam Medical Science
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• v.39 no.1
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• pp.24-30
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• 2022

Background: Texture analysis has been used as a method for quantifying image properties based on textural features. The aim of the present study was to evaluate the usefulness of magnetic resonance imaging (MRI) texture analysis for the evaluation of viable ovarian tissue on the perfusion map of ovarian endometriosis. Methods: To generate a normalized perfusion map, subtracted T1-weighted imaging (T1WI), T1WI and contrast-enhanced T1W1 with sequences were performed using the same parameters in 25 patients with surgically confirmed ovarian endometriosis. Integrated density is defined as the sum of the values of the pixels in the image or selection. We investigated the parameters for texture analysis in ovarian endometriosis, including angular second moment (ASM), contrast, correlation, inverse difference moment (IDM), and entropy, which is equivalent to the product of area and mean gray value. Results: The perfusion ratio and integrated density of normal ovary were 0.52±0.05 and 238.72±136.21, respectively. Compared with the normal ovary, the affected ovary showed significant differences in total size (p<0.001), fractional area ratio (p<0.001), and perfusion ratio (p=0.010) but no significant differences in perfused tissue area (p=0.158) and integrated density (p=0.112). In comparison of parameters for texture analysis between the ovary with endometriosis and the contralateral normal ovary, ASM (p=0.004), contrast (p=0.002), IDM (p<0.001), and entropy (p=0.028) showed significant differences. A linear regression analysis revealed that fractional area had significant correlations with ASM (r²=0.211), IDM (r²=0.332), and entropy (r²=0.289). Conclusion: MRI texture analysis could be useful for the evaluation of viable ovarian tissues in patients with ovarian endometriosis.