• Asian-Australasian Journal of Animal Sciences
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• v.16 no.1
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• pp.23-28
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• 2003

The present study was undertaken to investigate the effects of PVP concentration and exposure temperature to vitrification solution on the post-thaw survival, in vitro maturation and development of immature bovine oocytes (germinal vesicle stage). The vitrification solution (VS) consisted of 40% ethylene glycol (EG)+0.5 M sucrose (S)+10% FBS. PVP was added to VS: 0%, 5% or 10%. The cumulus-oocyte complexes (COCs) were diluted in VS as one step, after 2 min the COCs were loaded in straw and vitrified by direct immersion into liquid nitrogen. For thawing, the straws were plunged into $30^{\circ}C$ water bath for 10s. After thawing, the oocytes were diluted in 0.5 M (in DPBS with 10% FBS) sucrose solution for 5 min. The survival rate (FDA-test and trypan blue) of immature bovine oocytes was measured. The survival rate was higher in 5% PVP (91.5%) than in 0% (64.2%) or in 10% PVP (79.7%). The proportion of metaphase II formation was 69.35% in control (no vitrified COCs), 9.3% in 40% EG+0.5 M S+0% PVP and 21.05% in 40% EG+0.5 M S+5% PVP (p<0.05). The effect of room temperature ($25^{\circ}C$ for 10 min) and cold temperature ($4^{\circ}C$ for 10 min) on COCs were determined in this study. After IVF, the cleavage and blastocysts rate of oocytes exposed to room temperature and cold temperature in VS+5% PVP was significantly different (2 cell: 63.20% vs 37.97%, blastocysts: 18.40% vs 2.53%). The cleavage rates of frozen-thawed oocytes were 20.53% with PVP and 22.13% without PVP (p>0.05). Two out of 151 oocytes (1.32%) developed to blastocyst stage after frozen-thawed with 5% PVP (p>0.05). Development of oocytes after frozen-thawing to the 2 cell were not significantly affected with or without PVP following IVF. However, the vitrification of immature bovine oocytes with PVP maintained the ability to develop to the blastocyst stage after IVM-IVF and IVC, while no blastocysts were obtained from oocytes vitrified without PVP. These results suggested that PVP has a protective role for vitrification of immature bovine oocytes as far as survival is concerned, however, the protection was not sufficient enough to support blastocyst formation.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.6
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• pp.2527-2530
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• 2015

Background: The cancer of the prostate risk assessment (CAPRA) score has been defined to predict prostate cancer recurrence based on the pre-clinical data, then pathological data have also been incorporated. Thus, CAPRA post-surgical (CAPRA-S) score has been developed based on six criteria (prostate specific antigen (PSA) at diagnosis, pathological Gleason score, and information on surgical margin, seminal vesicle invasion, extracapsular extension and lymph node involvement) for the prediction of post-surgical recurrences. In the present study, biochemical recurrence (BCR)-free probabilities after open retropubic radical prostatectomy (RP) were evaluated by the CAPRA-S scoring system and its three-risk level model. Materials and Methods: CAPRA-S scores (0-12) of our 240 radical prostatectomies performed between January 2000-May 2011 were calculated. Patients were distributed into CAPRA-S score groups and also into three-risk groups as low, intermediate and high. BCR-free probabilities were assessed and compared using Kaplan-Meier analysis and Cox proportional hazards regression. Ability of CAPRA-S in BCR detection was evaluated by concordance index (c-index). Results: BCR was present in 41 of total 240 patients (17.1%) and the mean follow-up time was $51.7{\pm}33.0$ months. Mean BCR-free survival time was 98.3 months (95% CI: 92.3-104.2). Of the patients in low, intermediate and high risk groups, 5.4%, 22.0% and 58.8% had BCR, respectively and the difference among the three groups was significant (P = 0.0001). C-indices of CAPRA-S score and three-risk groups for detecting BCR-free probabilities in 5-yr were 0.87 and 0.81, respectively. Conclusions: Both CAPRA-S score and its three-risk level model well predicted BCR after RP with high c-index levels in our center. Therefore, it is a clinically reliable post-operative risk stratifier and disease recurrence predictor for prostate cancer.

• The Korean Journal of Pharmacology
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• v.24 no.1
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• pp.43-52
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• 1988

To elucidate the mechanism of action of local anesthetics, the effects of local anethetics on the microenvironment of the lipid bilayers of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine brain and dimyristoylphosphatidylcholine (DMPC) multilamellar liposomes were investigated employing the intermolecular excimer fluorescence technique and differential scanning calorimetry (DSC). The relative intensities of excimer and monomer fluorescence of pyrene are a simple linear function of the viscosity of a homologous series of solvents. The microviscosity(${\eta}$)of the hydrocarbon region of SPMV was measured by this method and the value was $57.3{\pm}5.3\;cP$ at $37^{\circ}C$. In the presence of lidocaine-HCl and procaine-HCl, the values decreased to $46.5{\pm}5.1\;cP$ and $54.7{\pm}4.8\;cP$, respectvely. The differential scanning thermograms of DMPC multilamellar liposomes showed that local anesthetics significantly lowered the phase transition temperature, broadened the thermogram peaks, and reduced the size of the cooperative unit. These results indicate that local anesthetics have significant fluidizing effects on biomembranes and perturbation of membrane lipids may produce some, but not all, of their pharmacological actions.

• The Korean Journal of Pharmacology
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• v.28 no.2
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• pp.191-199
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• 1992

Selective quenching of 1,6-diphenyl-1,3,5-hexatriene (DPH) by trinitrophenyl groups was utilized to examine the transbilayer fluidity asymmetry of model membranes of total lipids (SPMVTL) extracted from synaptosomal plasma membrane vesicles (SPMV). The polarization (P), anisotropy (r), limiting anisotropy $(r_{\infty})$, and order parameter (S) of DPH in the inner monolayer were 0.031, 0.025, 0.033, and 0.070, respectively, greater than calculated for the outer monolayer of SPMVTL. Selective quenching of DPH by trinitrophenyl groups was also utilized to examine the effects of n-alkanols on the individual monolayer structure of SPMVTL. n-Alkanols fluidized the hydrocarbon region of bulk SPMVTL, and the potencies of n-alkanols up to 1-nonanol increased with carbon chain length. It appears that the potencies in bilayer fluidization increase by 1 order of magnitude as the carbon chain length increases by two carbon atoms. The cut-off phenomenon was reached at 1-decanol, where further increase in hydrocarbon length resulted in a decrease in pharmacological activity. The n-alkanols had greater fluidizing effects on the outer monolayer as compared to the inner monolayer of SPMVTL, even though these selective effects tended to become weaker as carbon chain length increased. Thus, it has been proven that n-alkanols exhibit selective rather than nonselective fluidizing effects within transbilayer domains of SPMVTL.

• Journal of Korean Neurosurgical Society
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• v.63 no.5
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• pp.566-578
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• 2020

Objective : Radiation is known to induce autophagy in malignant glioma cells whether it is cytocidal or cytoprotective. Dexamethasone is frequently used to reduce tumor-associated brain edema, especially during radiation therapy. The purpose of the study was to determine whether and how dexamethasone affects autophagy in irradiated malignant glioma cells and to identify possible intervening molecular pathways. Methods : We prepared p53 mutant U373 and LN229 glioma cell lines, which varied by phosphatase and tensin homolog (PTEN) mutational status and were used to make U373 stable transfected cells expressing GFP-LC3 protein. After performing cell survival assay after irradiation, the IC₅₀ radiation dose was determined. Dexamethasone dose (10 μM) was determined from the literature and added to the glioma cells 24 hours before the irradiation. The effect of adding dexamethasone was evaluated by cell survival assay or clonogenic assay and cell cycle analysis. Measurement of autophagy was visualized by western blot of LC3-I/LC3-II and quantified by the GFP-LC3 punctuated pattern under fluorescence microscopy and acridine orange staining for acidic vesicle organelles by flow cytometry. Results : Dexamethasone increased cell survival in both U373 and LN229 cells after irradiation. It interfered with autophagy after irradiation differently depending on the PTEN mutational status : the autophagy decreased in U373 (PTEN-mutated) cells but increased in LN229 (PTEN wild-type) cells. Inhibition of protein kinase B (AKT) phosphorylation after irradiation by LY294002 reversed the dexamethasone-induced decrease of autophagy and cell death in U373 cells but provoked no effect on both autophagy and cell survival in LN229 cells. After ATG5 knockdown, radiation-induced autophagy decreased and the effect of dexamethasone also diminished in both cell lines. The diminished autophagy resulted in a partial reversal of dexamethasone protection from cell death after irradiation in U373 cells; however, no significant change was observed in surviving fraction LN229 cells. Conclusion : Dexamethasone increased cell survival in p53 mutated malignant glioma cells and increased autophagy in PTEN-mutant malignant glioma cell but not in PTEN-wildtype cell. The difference of autophagy response could be mediated though the phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin signaling pathway.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.3
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• pp.243-252
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• 1985

The mechanism of reproductive cycle of longchin goby Chasmichthys dolichognathus was investigated for the annual variations of gonadosomatic index(GSI), histological changes of gonad, reproductive cycle, and initiating and terminating factors of the breeding season. The experimental fish were captured in the tide pool of Tongbaeksom, Pusan, Korea from February 1983 to September 1984. Experimental fish for the study of initiating and terminating factors of the breeding season were maintained in filtered recirculating aquariums and exposed to various temperature and photoperiod regimes. The ovary is a pair of sac-shaped organ, and the testicular structure is tubular and it is connected to the seminal vesicle which is located at the posterior end of the testis. GSI began to increase from February when the water temperature began to increase and reached peak in April. It began to decrease from August, the top water temperature season, and maintained relatively low values until January. The annual reproductive cycle includes successive six developmental stages : early growing from December to February, late growing from January to March, early mature from February to May, late mature and ripe from March to August, spawning from April to July, degeneration and resting stage from May to December. An experimental study based on the reproductive cycle of this species indicated that rising temperature under long photoperiod during spring stimulated gonadal maturation, whereas terminating factor of the breeding season with recrudescence of the gonads was demonstrated as high water temperature during summer regardless of photoperiod.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.24 no.1
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• pp.67-77
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• 1994

The purpose of this study was to investigate the irradiatiion effects on the capillary and endothelial cell in the submandibular gland. Sprague-Dawley strain male rats were singly irradiated to their neck region with the dose of 5Gy by 6MV X-irradiation and sacrificed on the 6 hours, 12 hours, 1, 3, 7, and 14days after irradiation. The authors observed the histological changes of the capillary at H & E and PAS staining under a light microscope, and also observed the ultrastructural changes of the endothelial cell using a transmission electron microscope. The obtaining results were as follows: 1. In the light microscopic examination, the capillary density was slightly increased on the 1day after irradiation, and increased until the 7 days after irradiatiion. After then, capillary density was apparently decreased. 2. The reaction to PAS staining at acinar cells was decreased on the 6 hours after irradiation, and recovered on the 7days after irradiation. But reaction was decreased on the 14days after irradiation agan, after then, gradually recovered with days. 3. In the transmission electron microscopic examination, mild proliferation of cytoplasmic process of the endothelial cell and reduction in luminal size were observed just after irradiation. After then, nuclear degeneration, marked proliferation of cytoplasmic process, thickened basal lamina, and numerous cytoplasmic vesicles were observed on the 1day after irradiation. These changes were recovered to normal on the 14days after 5Gy group, but not with 10Gy irradiation group. And destruction of endothelial cell and loss of basal lamina were not observed in both groups. 4. From the above results, reduction in luminal size, proliferation of cytoplasmic process and thickening of basal lamina were observed as the irradiation effects on the capillary and endothelial cell of the submandibular gland. And also, these changes may induce increase in capillary number and endothelial permeability by means of increase of cytoplasmic vesicle formation. The changes appeared earlier and more prominent in 10Gy irradiated group than in 5Gy irradiated group.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.4
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• pp.193-202
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• 2011

Objective: We found previously that $interferon$ $regulatory$ factor ($Irf$)-1 is a germinal vesicle (GV)-selective gene that highly expressed in GV as compared to metaphase II oocytes. To our knowledge, the function of $Irf-1$ in oocytes has yet to be examined. The present study was conducted to determine the relationship between retinoic acid (RA) and RA-mediated expression of $Irf-1$ and the mouse oocyte maturation. Methods: Immature cumulus-oocyte-complexes (COCs) were collected from 17-day-old female mice and cultured $in$ $vitro$ for 16 hours in the presence of varying concentrations of RA (0-10 ${\mu}M$). Rate of oocyte maturation and activation was measured. Gene expression was measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and cytokine secretion in the medium was measured by Bio-Plex analysis. Apoptosis was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: The rates of oocyte maturation to metaphase II and oocyte activation increased significantly with RA treatment (10 nM-1 ${\mu}M$). With 100 nM RA treatment, lowest level of $Irf-1$ mRNA and cumulus cell's apoptosis was found. Among 23 cytokines measured by Bio-Plex system, the substantial changes in secretion of tumor necrosis factor-${\alpha}$, macrophage inflammatory protein-$1{\beta}$, eotaxin and interleukin-12 (p40) from COCs in response to RA were detected. Conclusion: We concluded that the maturation of oocytes and $Irf-1$ expression are negatively correlated, and RA enhances the developmental competence of mouse immature oocytes $in$ $vitro$ by suppressing apoptosis of cumulus cells. Using a mouse model, results of the present study provide insights into improved culture conditions for $in$ $vitro$ oocyte maturation and relevant cytokine production and secretion in assisted reproductive technology.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.4
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• pp.166-172
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• 2019

Objective: In vitro maturation (IVM) of immature oocytes can be useful for some infertile patients. In IVM programs, the rates of embryo formation and pregnancy are low. Therefore, it is essential to recognize the main factors involved in regulating oocyte maturation in vitro. The purpose of this study was to investigate the effects of growth differentiation factor 9 (GDF9) and cumulus cell (CC) supplementation in IVM medium on the rates of embryo formation and viability of human blastocysts. Methods: A total of 80 germinal vesicle oocytes from stimulated cycles underwent an IVM program. The oocytes were divided into four groups, where group I consisted of IVM media only and served as the control, group II consisted of IVM+CCs, group III consisted of IVM+GDF9 (200 ng/mL), and group IV consisted of IVM+CCs+GDF9 (200 ng/mL). Intracytoplasmic sperm injection was performed on the IVM oocytes, and the cleavage embryos that were generated were vitrified. Following thawing, the embryos were cultured for 3 additional days, and the viability rates of the developed blastocysts were determined. Results: The maturation rate of the oocytes did not differ significantly across the four groups. The fertilization rate in group II was significantly higher than that in the control group (76.5% vs. 46.2%). Embryo formation was significantly more frequent in all experimental groups than in the control group, while blastocyst formation did not show significant differences in the three experimental groups compared to the control. The mean viability rates in groups II, III, and IV were 58.16%, 55.91%, and 55.95%, respectively, versus 37.78% in the control group (p< 0.05). Conclusion: Supplementation of IVM culture media with GDF9 and CCs enhanced the fertilization, embryo formation, and viability rates of blastocysts generated from vitrified cleavage embryos.

• Journal of Aquaculture
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• v.9 no.4
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• pp.419-427
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• 1996

In order to obtain the basic information for seedling production of surf clam, Spisula sachalinensis, spawning induction and egg development were investigated. $NH_4OH$ addition and serotonin injection could induce the spawning in surf clam. Water temperature rising, sperm suspension immersion, UV-ray irradiated seawater and $H_2O_2$ addition less affected on induction of spawning than $NH_4OH$ and serotonin did. On the other hand, males were more sensitive to the treatments than females. The response time to initial spawning in the case of $NH_4OH$ addition was $3\~4$ hours. However in the case of serotonin injection, it was within 5 minutes. The number of eggs released by $NH_4OH$ addition were significantly more than those released by serotonin injection. The serotonin injection induced higher rates of germinal vesicle breakdown than the $NH_4OH$ addition. Fertilizing and hatching rates of the eggs also were the similar results. Eggs of surf clam were demersal isolated eggs and averaging $65.2{\pm}±1.8\;{\mu}m$ in diameter after spawning. Optimum range of water temperature for the development of egg was $15\~20^{\circ}C$, The required time for development of D-shaped larvae was 42 hours at $15^{\circ}C$ and 27 hours at $20^{\circ}C$, respectively.