• 한국발생생물학회지:발생과생식
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• 제22권4호
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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.

• 한국발생생물학회지:발생과생식
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• 제23권1호
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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).

• 한국발생생물학회지:발생과생식
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• 제23권1호
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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.

• 한국발생생물학회지:발생과생식
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• 제25권4호
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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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• 제57권1호
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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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• 제29권6호
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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.

• 한국발생생물학회지:발생과생식
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• 제22권4호
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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.

• 한국미생물·생명공학회지
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• 제50권1호
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• pp.63-70
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• 2022

Two small cryptic plasmids, pHG1 and pHG2, were isolated from Levilactobacillus zymae (formerly Lactobacillus zymae) GU240 and characterized. pHG1 is 1,814 bp in size with a GC content of 37.4% and contains two open reading frames. orf1 can potentially encode a protein of 101 amino acids (aa) with 99% identity with the copy number control protein of Lacticaseibacillus paracasei. orf2 can potentially encode a protein of 230 aa with 99% identity with a replication protein from multiple species. Six inverted repeats (IR I-VI) and six direct repeats (DR I-VI) were found in pHG1. pHG2 is 2,864 bp in size, with a GC content of 39.6%. pHG2 has two orfs. orf1 might encode a protein with 99% identity with the TrsL transmembrane protein. orf2 might encode a protein with 99% identity with plasmid recombination proteins from lactic acid bacteria. Both pHG1 and pHG2 may be useful as frames for constructing lactic acid bacteria-Escherichia coli shuttle vectors.

• Archives of Plastic Surgery
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• 제49권5호
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• pp.617-632
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• 2022

Tracheoesophageal puncture (TEP) and voice prosthesis insertion following laryngectomy may fail to form an adequate seal. When spontaneous closure of the fistula tract does not occur after conservative measures, surgical closure is required. The purpose of this study was to summarize the available evidence on surgical methods for TEP site closure. A comprehensive search across PubMed, Web of Science, SCOPUS, and Cochrane was performed to identify studies describing surgical techniques, outcomes, and complications for TEP closure. We evaluated the rate of unsuccessful TEP closure after surgical management. A meta-analysis with a random-effect method was performed. Thirty-four studies reporting on 144 patients satisfied inclusion criteria. The overall incidence of an unsuccessful TEP surgical closure was 6% (95% confidence interval [CI] 1-13%). Subgroup analysis showed an unsuccessful TEP closure rate for silicone button of 8% (95% CI < 1-43%), 7% (95% CI < 1-34%) for dermal graft interposition, < 1% (95% CI < 1-37%) for radial forearm free flap, < 1% (95% CI < 1-52%) for ligation of the fistula, 17% (95% CI < 1-64%) for interposition of a deltopectoral flap, 9% (95% CI < 1-28%) for primary closure, and 2% (95% CI < 1-20%) for interposition of a sternocleidomastoid muscle flap. Critical assessment of the reconstructive modality should take into consideration previous history of surgery or radiotherapy. Nonirradiated fields and small defects may benefit from fistula excision and tracheal and esophageal multilayer closure. In cases of previous radiotherapy, local flaps or free tissue transfer yield high successful TEP closure rates. Depending on the defect size, sternocleidomastoid muscle flap or fasciocutaneous free flaps are optimal alternatives.

• 한국발생생물학회지:발생과생식
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• 제24권4호
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• pp.249-261
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• 2020

Spermatogonial stem cells (SSCs) have stemness characteristics, including germ cell-specific imprints that allow them to form gametes. Spermatogenesis involves changes in gene expression such as a transition from expression of somatic to germ cell-specific genes, global repression of gene expression, meiotic sex chromosome inactivation, highly condensed packing of the nucleus with protamines, and morphogenesis. These step-by-step processes finally generate spermatozoa that are fertilization competent. Dynamic epigenetic modifications also confer totipotency to germ cells after fertilization. Primordial germ cells (PGCs) in embryos do not enter meiosis, remain in the proliferative stage, and are referred to as gonocytes, before entering quiescence. Gonocytes develop into SSCs at about 6 days after birth in rodents. Although chromatin structural modification by Polycomb is essential for gene silencing in mammals, and epigenetic changes are critical in spermatogenesis, a comprehensive understanding of transcriptional regulation is lacking. Recently, we evaluated the expression profiles of Yin Yang 1 (YY1) and CP2c in the gonads of E14.5 and 12-week-old mice. YY1 localizes at the nucleus and/or cytoplasm at specific stages of spermatogenesis, possibly by interaction with CP2c and YY1-interacting transcription factor. In the present article, we discuss the possible roles of YY1 and CP2c in spermatogenesis and stemness based on our results and a review of the relevant literature.