• Development and Reproduction
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• v.22 no.1
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• pp.39-53
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• 2018

The purpose of this study is to investigate the early life history of the Rhodeus fish, Rhodeus uyekii and R. ocellatus, in the Nakdong River to use as the preliminary data for the systematic study. The embryos used in the study were fertilized eggs (embryo) and larvae after artificial fertilization. The long diameter of the eggs of the R. uyekii was 3.39-3.97 mm (average $3.68{\pm}0.41mm$, n=30) and the short diameter was 1.36-1.55 mm (average $1.45{\pm}0.13mm$, n=30). The long diameter of the eggs of the R. ocellatus was 2.53-2.71 mm (average $2.62{\pm}0.12mm$, n=30) and the short diameter was 1.47-1.60 mm (average $1.53{\pm}0.09mm$, n=30). Hatching time was 48 hours for the R. uyekii and 50 hours for the R. ocellatus given that the average water temperature was $21.5^{\circ}C$. The hatched larvae were 4.95-5.00 mm (average $4.98{\pm}0.04mm$, n=5) for the R. uyekii and the total length was 3.66-3.69 mm (average $3.67{\pm}0.02mm$, n=5) for the R. ocellatus. R. uyekii was found to be 15.5-15.8 mm at total length (average $15.6{\pm}0.21mm$, n=5) on the 56 days after hatching with the number of dorsal fins being ⅲ-9, anal fins ⅲ-10, ventral fins ⅲ-5. The R. ocellatus was found to be 15.8-16.0 mm (average $15.9{\pm}0.14mm$, n=5) at total length on the 58 days with the number of dorsal fins being ⅲ-11, anal fins ⅲ-12 and ventral fins ⅲ-5 where the number of all fin stalks reached maximum.

• Development and Reproduction
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• v.24 no.3
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• pp.215-224
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• 2020

Seawater adaptability of steelhead trout increases along with the increase in the size of the fish, independent of parr-smolt transformation. Three 96 h seawater challenge tests were conducted to determine the size at which seawater adaptability of steelhead trout develops. Plasma Na⁺ and Cl^- levels, moisture content, gill Na⁺/K⁺ ATPase activity, and mortality during the 96 h after direct transfer to seawater (32 ppt) were determined. Plasma Na⁺ and Cl^- levels in 50 g fish continuously increased during the 96 h after the transfer to seawater (p<0.05), but the levels in 100 and 150 g fish leveled off after 24 h (p<0.05). Both 100 and 150 g size steelhead trout maintained muscle moisture content (%) better than 50 g size fish (p<0.05). Gill Na⁺/K⁺ ATPase activity in the 100 g size group increased in a time-dependent manner after transfer to seawater (p<0.05), whereas activity in the 50 and 150 g sizes did not increase (p>0.05), for which a possible explanation was discussed. A mere 2.6% mortality in both the 50 and 150 g size groups was observed. In conclusion, the current results indicate that 50 g size steelhead trout did not show development of a high level of hypoosmoregulatory capacity, whereas fish in the 100 and 150 g size groups showed a high level in our experimental conditions. Therefore, the steelhead trout larger than a 100 g size is recommended for transfer to seawater culture.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.1
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• pp.45-54
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• 2005

이 연구는 SKBR3 인간 유방암세포주에 대한 귀전우 메탄올추출물(CME)의 증식억제 효과, 세포사 유발 효과 및 항산화 활성을 확인하기 위해 이루어졌다. SKBR3 유방암세포주는 48시간동안 다양한 농도($0{\sim}20{\mu}g/m{\ell}$)의 CME가 제공된 곳에서 배양되었고, MMT 측정법을 이용하여 세포생존율을 평가하였다. CME의 50%에서 효과를 나타내게 하는 약물농도인 $ED_{50}$ (effective dose 50%)은 $6.5{\pm}0.3{\mu}g/m{\ell}$) 이며, 투여량이 증가함에 따라 농도에 의존하여 세포증식이 억제되는 것으로 나타났다. 또한 CME의 증식억제 효과는 유방암세포주의 세포사와 관련됨의 세포의 형태학적 변화와 올리고뉴클레오솜 DNA 파편의 확인을 통해 알 수 있었다. 또한 다양한 농도와 배양시간에서 CME가 ROS의 생산을 억제한다는 것을 확인할 수 있었다. 이런 결과들은 귀전우의 메탄올추출물이 SKBR3 인간 유방암세포주에 대해 강력한 증식억제 효과와 강한 항산화 효과를 나타낼 뿐만 아니라 세포사를 유도하는 효능을 가지고 있음을 시사한다. 이러한 효능은 약물에 대한 노출시간과 투여량에 의존하였다. 따라서 귀전우는 다양한 기전에 의해 유방암 세포에 대한 억제효과를 가질 수 있을 것으로 인식할 수 있다.

• Development and Reproduction
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• v.22 no.4
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• pp.297-307
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• 2018

The objective of this study was to evaluate the effects of alpha-linolenic acid (ALA) during in vitro maturation (IVM) on cumulus expansion, nuclear maturation, fertilization capacity and subsequent development in porcine oocytes. The oocytes were incubated with 0, 25, 50, and $100{\mu}M$ ALA. Cumulus expansion was measured at 22 h, and gene expresison and nuclear maturation were analyzed at 44 h after maturation. Then, mature oocytes with ALA were inseminated, and fertilization parameters and embryo development were evaluated. In results, both of cumulus expansion and nuclear maturation were increased in $50{\mu}M$ ALA groups compared to control groups (p<0.05). However, expression of gap junction protein alpha 1 (GJA1, cumulus expansion-related gene), delta-6 desaturase (FADS1, fatty acid metabolism-related gene), and delta-5 desaturase (FADS2) mRNA in cumulus cells were reduced by $50{\mu}M$ ALA treatment (p<0.05). Cleavage rate was enhanced in 25 and $50{\mu}M$ ALA groups (p<0.05), especially, treatment of $50{\mu}M$ ALA promoted early embryo develop to 4 and 8 cell stages (p<0.05). However, blastocyst formation and number of cells in blastocyst were not differ in 25 and $50{\mu}M$ ALA groups. Our findings show that ALA treatment during maturation could improve nuclear maturation, fertilization, and early embryo development through enhancing of cumulus expansion, however, fatty acid metabolism- and cumulus expansion-related genes were down-regulated. Therefore, addition of ALA during IVM of oocytes could improve fertilization and developmental competence, and further studies regarding with the mechanism of ALA metabolism are needed.

• Development and Reproduction
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• v.23 no.1
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• pp.21-34
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• 2019

The effects of the anaesthetic agents, clove oil and mixture of clove oil with lidocaine-HCl were evaluated on river puffer, Takifugu obscurus and tiger puffer, T. rubripes. Anaesthesia times of clove oil were affected by water temperature ($20^{\circ}C$, $24^{\circ}C$, and $28^{\circ}C$) and salinity (10, 20, and 30 ppt). Anaesthesia times of mixed samples were significantly similar with regard to exposure and recovery times, and all samples satisfied anaesthesia criteria (exposure time within 3 min and recovery time within 5 min) under the various temperatures and salinities, and the lowest to highest concentration of anaesthetics (p<0.05). Both species river puffer and tiger puffer had short exposure time with a high anaesthesia dose, high temperature ($28^{\circ}C$) and intermediate salinity (20 ppt), and were highly affected by temperature and salinity (p<0.05). The mixed anaesthetics had rapid exposure times and long recovery times in contrast to the effects of clove oil. Cortisol concentrations under the conditions of various clove oil dosages, salinity, and temperature for both species increased until 12 hrs after recovery from anaesthesia (p<0.05). After 12 hrs, cortisol concentrations decreased until after 48 hrs (p<0.05). During the simulated transportation of both species, control and sedated clove oil groups (5 ppm) were measured for water parameters, dissolved oxygen (DO), $CO_2$, respiratory frequency, $NH_4{^+}$, and pH for 6 hrs in 1 hr intervals. Water parameters of sedated groups and controls were significantly different after 2 hrs (p<0.05).

• Development and Reproduction
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• v.23 no.1
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• pp.43-54
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• 2019

We examined the effects of endoplasmic reticulum (ER) stress inhibitor treatment during the micromanipulation of porcine somatic cell nuclear transfer (SCNT) on the in vitro development of SCNT embryos. ER stress inhibitors such as salubrinal (200 nM) and tauroursodeoxycholic acid (TUDCA; $100{\mu}M$) were added to the micromanipulation medium and holding medium. The expression of X-box binding protein 1 (Xbp1), ER-stress-associated genes, and apoptotic genes in SCNT embryos was confirmed at the one-cell and blastocyst stages. Levels of Xbp1 splicing and expression of ER-stress-associated genes in SCNT embryos at the one-cell stage decreased significantly with TUDCA treatment (p<0.05). The expression of ER-stress-associated genes also decreased slightly with the addition of both salubrinal and TUDCA (Sal+TUD). The expression levels of caspase-3 and Bcl2-associated X protein (Bax) mRNA were also significantly lower in the TUDCA and Sal+TUD treatments (p<0.05). At the blastocyst stage, there were no differences in levels of Xbp1 splicing, and transcription of ER-stress-associated genes and apoptosis genes between control and treatment groups. However, the blastocyst formation rate (20.2%) and mean blastocyst cell number ($63.0{\pm}7.2$) were significantly higher (p<0.05) for embryos in the TUDCA treatment compared with those for control (12.6% and $41.7{\pm}3.1$, respectively). These results indicate that the addition of ER-stress inhibitors, especially TUDCA, during micromanipulation can inhibit cellular damage and enhance in vitro development of SCNT embryos by reducing stress levels in the ER.

• Development and Reproduction
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• v.25 no.4
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• pp.199-211
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• 2021

Equine chorionic gonadotropin (eCG), produced by the endometrial cups of the placenta after the first trimester, is a specific glycoprotein that displays dual luteinizing hormone (LH)-like and follicle-stimulating hormone (FSH)-like effects in non-equid species. However, in equidaes, eCG exhibits only LH-like activity. To identify the specific biological functions of glycosylated sites in eCG, we constructed the following site mutants of N- and O-linked glycosylation: eCGβ/αΔ56, substitution of α-subunit⁵⁶ N-linked glycosylation site; eCGβ-D/α, deletion of the O-linked glycosylation sites at the β-subunit, and eCGβ-D/αΔ56, double mutant. We produced recombinant eCG (rec-eCG) proteins in Chinese hamster ovary suspension (CHO-S) cells. We examined the biological activity of rec-eCG proteins in CHO-K1 cells expressing the eLH/CG receptor and found that signal transduction activities of deglycosylated mutants remarkably decreased. The EC₅₀ levels of eCGβ/αΔ56, eCGβ-D/α, and eCGβ-D/αΔ56 mutants decreased by 2.1-, 5.6-, and 3.4-fold, respectively, compared to that of wild-type eCG. The Rmax values of the mutants were 56%-80% those of wild-type eCG (141.9 nmol/10⁴ cells). Our results indicate that the biological activity of eCG is greatly affected by the removal of N- and O-linked glycosylation sites in cells expressing eLH/CGR. These results provide important information on rec-eCG in the regulation of specific glycosylation sites and improve our understanding of the specific biological activity of rec-eCG glycosylation sites in equidaes.

• Parasites, Hosts and Diseases
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• v.57 no.1
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• pp.33-38
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• 2019

Trichomoniasis is a common sexually transmitted infection caused by Trichomonas vaginalis, which actually does not exist a vaccine for control or prevention. Thus, the identification of new and potent immunogens in T. vaginalis, which can contribute to the development of a vaccine against this parasite, is necessary. Therefore, the aim of this work was to evaluate the potential of a recombinant Transient Receptor Potential-like channel of T. vaginalis (TvTRPV), as a promising immunogen in BALB/c mice. First, TvTRPV was cloned and expressed as a recombinant protein in Escherichia coli BL21 cells and purified by nickel affinity. Next, BALB/c mice were immunized and the antibody levels in mice serum and cytokines from the supernatant of macrophages and from co-culture systems were evaluated. Recombinant TvTRPV triggered high levels of specific total IgG in sera from the immunized mice. Also, a statistically significant increase of cytokines: $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ after stimulation with the corresponding antigens in vitro, was identified. Moreover, co-cultures using $CD4^+$ T cells from immunized mice were able to identify higher levels of IL-10 and $IFN-{\gamma}$. These results were useful to validate the immunogenicity of TvTRPV in BALB/c mice, where IL-10-$IFN-{\gamma}$-secreting cells could play a role in infection control, supporting the potential of TvTRPV as a promising target for vaccine against T. vaginalis.

• Journal of Gynecologic Oncology
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• v.29 no.6
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• pp.92.1-92.9
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• 2018

Objective: To evaluate the effectiveness of the pulmonary recruitment maneuver (PRM) at the end of the operation to decrease laparoscopy-induced abdominal or shoulder pain after gynecological oncologic surgery. Methods: In total, 113 women undergoing laparoscopic surgery for malignant or premalignant gynecological lesions were assigned randomly to two groups: the PRM group (the patient was placed in the Trendelenburg position ($30^{\circ}$) and the PRM, consisting of two manual pulmonary inflations to a maximum pressure of $40cmH_2O$) (n=54) and the control group (n=52). Postoperative shoulder and abdominal pain was assessed 12, 24, and 48 hours later using a visual analog scale (0-10). In addition, the incidence of post-discharge nausea and vomiting was recorded until 48 hours after discharge. Results: Postoperative shoulder pain at 12 and 24 hours was significantly less severe in the PRM group ($2.2{\pm}0.5$ and $2.0{\pm}0.4$) than in the control group ($4.0{\pm}0.5$ and $3.9{\pm}0.4$; both p<0.001). The PRM significantly reduced the severity of upper abdominal pain at 12 and 24 h compared with the control group ($3.1{\pm}0.4$ and $2.9{\pm}0.4$ vs. $2.9{\pm}0.5$ and $4.9{\pm}0.5$; both p<0.001). The analgesic requirement during the postoperative period was similar in the two groups (control group, 78.8%; PRM group, 75.9%; p=0.719). Conclusion: The PRM effectively and safely reduced postoperative shoulder and upper abdominal pain levels in patients undergoing laparoscopic gynecological oncologic surgery. Trial registry at ClinicalTrials.gov, NCT01940042.

• Development and Reproduction
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• v.22 no.4
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• pp.369-378
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• 2018

We determined the morphologic characteristics (body weight and degree of abdomen inflation) of the red spotted grouper, Epinephelus akaara, mother fish producing healthy eggs. Experimental fish were chosen from fish reared in a sea cage. The fish were divided into four size groups by body weight: 400~600, 600~800, 800~1,000, and 1,000~1,200 g and four stages (I~IV) of the degree of abdomen inflation. After hormone treatment, we observed the amount of ovulation-induced eggs, and rates of buoyancy, fertilization, embryonic survival, and hatching. As a result, mother fish with a body weight of 600 g or more spawned, and the fertilization rate, embryonic survival rate, and hatching rate were high in the 800~1,000 g range, thus showing effective ovulation induction. As a result of dividing the degree of abdomen inflation based on the anal fin of the mother fish into I-IV stages and determining hormone treatment time, the GSI was $0.9{\pm}0.2%$ at stage I, $2.3{\pm}0.2%$ at stage II, $5.6{\pm0.2%$ at stage III, and $7.9{\pm}0.9%$ at stage IV. The flotation rate and hatching rate were highest at stage III, and the fertilization rate and embryonic survival rate were highest at stage IV. Therefore, in terms of egg quality, the amount of eggs collected per mother fish, maturation, and histology were different depending on the degree of abdomen inflation. At stage III, where the abdomen inflation degree of the mother fish was based on the basal part of the dorsal fin relative to the height of the anal fin was 1, the egg quality was highest.