• Development and Reproduction
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• v.27 no.1
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• pp.25-37
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• 2023

Acetaminophen [Paracetamol, N-acetyl-para-aminophenol (APAP)] is a common over-the-counter analgesic agent as nonsteroidal anti-inflammatory drugs (NSAIDs). The high doses or the long-term treatment of acetaminophen via usual gavage feeding resulted in damage of testicles that presented recoverable impairment, as well as liver and kidney. The influence of acetaminophen was examined in male golden hamsters treated with acetaminophen-containing diet feeding. They were divided into 5 groups and subjected to this experiment for 4 weeks: animals housed in long photoperiod (LP) as LP control, animals housed in short photoperiod (SP) for 4 weeks as SP control (SP4), and groups of animals treated with low, middle, and high concentrations of acetaminophen (Low, Middle, High groups). Also animals housed in SP for 8 weeks were included (SP8) to contrast testicular activities, if necessary. As results, spermatozoa filled the seminiferous tubules of the testicles of animals in LP control and SP4 groups. The aspects were seen in the animals taken diets of low and middle doses of acetaminophen. The animals who fed high dose of acetaminophen showed large or small testicles. The large testicles displayed all germ cells at the steps of spermatogenesis. The small testicles presented no sperm as the animals housed in SP for 8 weeks. Thus these results indicate that acetaminophen invokes the antigonadal effects and accelerates the regressing process of the testicles in the animals compared to the animals exposed to SP.

• Journal of Animal Reproduction and Biotechnology
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• v.38 no.4
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• pp.199-208
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• 2023

Background: Canine induced pluripotent stem cells (iPSCs) are an attractive source for veterinary regenerative medicine, disease modeling, and drug development. Here we used vitamin C (Vc) to improve the reprogramming efficiency of canine iPSCs, and its functions in the reprogramming process were elucidated. Methods: Retroviral transduction of Oct4, Sox2, Klf4, c-Myc (OSKM), and GFP was employed to induce reprogramming in canine fetal fibroblasts. Following transduction, the culture medium was subsequently replaced with ESC medium containing Vc to determine the effect on reprogramming activity. Results: The number of AP-positive iPSC colonies dramatically increased in culture conditions supplemented with Vc. Vc enhanced the efficacy of retrovirus transduction, which appears to be correlated with enhanced cell proliferation capacity. To confirm the characteristics of the Vc-treated iPSCs, the cells were cultured to passage 5, and pluripotency markers including Oct4, Sox2, Nanog, and Tra-1-60 were observed by immunocytochemistry. The expression of endogenous pluripotent genes (Oct4, Nanog, Rex1, and telomerase) were also verified by PCR. The complete silencing of exogenously transduced human OSKM factors was observed exclusively in canine iPSCs treated with Vc. Canine iPSCs treated with Vc are capable of forming embryoid bodies in vitro and have spontaneously differentiated into three germ layers. Conclusions: Our findings emphasize a straightforward method for enhancing the efficiency of canine iPSC generation and provide insight into the Vc effect on the reprogramming process.

• Reproductive and Developmental Biology
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• v.29 no.1
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• pp.1-7
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• 2005

Telomeres consisting of (TTAGGG)n tandem repeat DNA sequences and associated proteins are essential for chromosome stability and related with cell senescence, apoptosis and cancer. The telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. This study was carried out to identify the distribution of telomeres on mouse chromosomes and also to analyze the amount of telomeres and telomerase activity of mouse embryos at early embryonic stages. Germ cells and early embryos from 1 cell to blastocyst stage were analyzed. The amount of telomeres was analyzed by quantitative fluorescence in situ hybridization technique(Q-FISH) using a human telomeric DNA probe, and telomerase activity was measured by telomeric repeat amplification protocol assay(TRAP). In results, the telomeres on mouse chromosomes were distributed at the ends of all autosomes and sex chromosomes. Although the quantity of telomeres varied among chromosomes, most of chromosomes had higher amount in q-arm telomeres than in p-arm telomeres. The results of Q-FISH indicated that the relative amount of telomeres of mouse embryos in each embryonic stage was approximately the same except the higher amount in blastocysts. Using TRAP assay on mouse embryos, telomerase activity was detected in all preimplantation stages from mature oocytes to blastocysts. Especially the telomerase activity was significantly increased at the morula and blastocyst stage. In conclusion, there may be a close association between the amount of telomeres and telomerase activity in early embryonic stages, and analysis of telomere quantity and telomerase activity on early development will be helpful for the investigation of embryogenesis and embryonic cell differentiation in mice.

• IMMUNE NETWORK
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• v.4 no.4
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• pp.216-223
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• 2004

Background: It is well known that IgA isotype switching is induced by $TGF-{\beta}1$. LPS-activated mouse normal B cells well differentiate into IgA secreting plasma cells under the influence of $TGF-{\beta}1$. Nevertheless, there are lots of difficulties in studying normal B cells in detail because it is not simple to obtain highly purified B cells, showing low reproducibility and transfection efficacy, moreover impossible to keep continuous culture. To overcome these obstacles, it is desperately needed to develop B cell line which acts like normal B cells. In the present study, we investigated whether CH12F3-2A lymphoma cells are appropriate for studying IgA isotype switching event. Methods: CH12F3-2A B cell line was treated with LPS and $TGF-{\beta}1$, then levels of germ-line (GL) transcripts were measured by RT-PCR, and $GL{\alpha}$ promoter activity was measured by luciferase assay. In addition, membrane IgA (mIgA) expression and IgA secretion were determined by FACS and ELISA, respectively. Results: $TGF-{\beta}1$, regardless of the presence of LPS, increased level of $GL{\alpha}$ transcripts but not $GL{\gamma}2b$ transcripts. However, IgA secretion was increased dramatically by co-stimulation of LPS and $TGF-{\beta}1$. Both mIgA and IgA secretion in the presence of $TGF-{\beta}1$ were further increased by over-expression of Smad3/4. Finally, $GL{\alpha}$ promoter activity was increased by $TGF-{\beta}1$. Conclusion: CH12F3-2A cell line acts quite similarly to the normal B cells which have been previously reported regarding IgA expression. Thus, CH12F3-2A lymphoma cell line appears to be adequate for the investigation of the mechanism(s) of IgA isotype switching at the cellular and molecular levels.

• Journal of the Korean Society of Food Culture
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• v.20 no.2
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• pp.186-193
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• 2005

The antioxidative and antimicrobial activities were carried on the leeks (Allium tuberosun R.) ethanol extracts in order to find out new food functional components. Three species of leeks used in this study were Chinese leek(long type, LL), general leek(medium type, LM), and medicinal leek (short type, LS). Total amounts of polyphenol compounds in LS was shown as the highest (436.60mg%) value. All of ethanol extracts of these leeks were shown to be had good electron donating ability(EDA) and nitrite scavenging activity. Specialty, the ethanol extract of LS(LSEx) had the highest EDA 30.47% and nitrite scavenging activity 77.24% and the lowest was LMEx. The antioxidative activities of these ethanol extracts on the corn germ oil were measured by peroxide values(POV) and conjugated diene values (CDV) storaging for 30 days at $60{\pm}2^{\circ}C$. The antioxidative activities of these extracts by POV and CDV were determined as following order as LSEx> LLEx) LMEx. The antioxidative activities of all extract were presented as high tendency by increasing adding amounts (0.02%>0.05%>0.1%). When the antioxldative activities were compared with BHT and ${\alpha}-tocopherol$, the degree of the antioxidative activities of these extracts were certified as lower than BHT and higher than ${\alpha}-tocopherol$. And also LSEx, LMEx, and LLEx had antimicrobial effects on the several micro organisms, especially the effect on the Pseudomonas aeruginasa was remarkable. While LMEx had shown inhibit effect on most of micro organism used in this study.

• Molecules and Cells
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• v.22 no.3
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• pp.343-352
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• 2006

Sialic acid is a sugar typically found at the N-glycan termini of glycoproteins in mammalian cells. Lec3 CHO cell mutants are deficient in epimerase activity, due to a defect in the gene that encodes a bifunctional UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE). Sialic acid modification on the cell surface is partially affected in these cells. We have mutagenized Lec3 CHO cells and isolated six mutants (termed C2m) deficient in the cell surface expression of polysialic acid (PSA). Mutant C2m9 was partially defective in expression of cell-surface PSA and wheat germ agglutinin (WGA) binding, while in the other five mutants, both cell-surface PSA and WGA binding were undetectable. PSA expression was restored by complementation with the gene encoding the CMP-sialic acid transporter (CST), indicating that CST mutations were responsible for the phenotypes of the C2m cells. We characterized the CST mutations in these cells by Northern blotting and RT-PCR. C2m9 and C2m45 carried missense mutations resulting in glycine to glutamate substitutions at amino acids 217 (G217E) and 256 (G256E), respectively. C2m13, C2m39 and C2m31 had nonsense mutations that resulted in decreased CST mRNA stability, and C2m34 carried a putative splice site mutation. PSA and CD15s expression in CST-deficient Lec2 cells were partially rescued by G217E CST, but not by G256E CST, although both proteins were expressed at similar levels, and localized to the Golgi. These results indicate that the novel missense mutations isolated in this study affect CST activity.

• Journal of the Korean Society of Food Culture
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• v.19 no.2
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• pp.170-176
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• 2004

The antioxidative and antimicrobial activity were carried on the Chopi(Zanthoxylum pipperitum) extracts by six kinds of solvents in order to find out new natural food additives. Six solvents were used methanol(MeOH), n-hexan(hexane), chloroform$(CHCl_3)$, ethylacetate(EtOAc), buthanol(BuOH), and water(water) and methanol extract(MEex), hexan extract(HEex), chloroform extract(CHex), ethylacetate extract(EAex), and buthanol extract(BUex), water extract(WAex). The antioxdative activities of them extracts were determined by peroxide value(POV), conjugated diene value(CDV) of corn germ oil storaged for 30 days at $60{\pm}2^{\circ}C$. These extracts were added as 0.02, 0.1, 0.2% of each extracts and compared with ${\alpha}-tocopherol$, and BHT. The antioxidative activities of 0.02% extract were as follows in decreasing order BUex > WAex, BHT, MEex > HEex, EAex, CHex > TOC, and control. While, BUex and CHex among these extracts were shown to be had antimicrobial effects on the microorganism such as Escheria coli, Salmonella typhimurium, Psedomonas aeruginosa, Staphylococcus aureus, and Listeria monocytogenes. Finally to find out the preference of chopi we had made Yugwanzageon (a kind of shallow fried meat ball)by adding chopi powder. The result were similar to control(not added chopi powder) in case of 0.1% chopi adding Yugwanzageon.(p<0.05) Therefore it was thought to be possible that chopi powder was used Yugwanzageon preperation.

• Development and Reproduction
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• v.6 no.1
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• pp.55-66
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• 2002

Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.7
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• pp.1003-1007
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• 2013

In this study, the antibacterial activities of selected barleys (UB, unhulled barley; PB, pearl barley; and NB, naked barley) and wheat (WG, wheat with germ and endosperm) extracts were evaluated against the food-borne pathogens Staphylococcus aureus KCTC 1927, Escherichia coli KCTC 2593, Salmonella Typhimurium KCTC 2054, and Bacillus cereus KCTC 1014. The amount of the antibacterial biomarker, 2,6-dimethoxy-1,4-benzoquinone (DMBQ), present in selected barleys and wheat, was measured by HPLC. Furthermore, antioxidant activity of samples was determined using the oxygen radical absorbance capacity (ORAC) assay. WG ($22.35{\pm}0.04mm$) was found to be highly inhibitory to Staphylococcus aureus followed by UB ($17.91{\pm}0.10mm$), PB ($16.87{\pm}0.05mm$), and NB ($15.69{\pm}0.20mm$). The antibacterial activity of the selected grains was correlated with antioxidant activities and the amount of DMBQ (Pearson's correlation coefficient, 0.7831). The antioxidant activity of the selected grains was also correlated with the total phenolic content (Pearson's correlation coefficient, 0.9934). WG extract showed significantly higher antibacterial activity, compared with barley extracts such as UB, PB, and NB. The results of this study suggest that barley has a potential in the development of natural antimicrobials and food preservatives for controlling food-borne pathogens.

• Development and Reproduction
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• v.4 no.1
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• pp.53-59
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• 2000

Endocrine disruptors (EDs) are exogenous chemicals which interfere several aspects of natural hormone properties. EDs with estrogenic activity have been recently reported to cause animal reproductive problems. This study was performed to investigate the effects of bisphenol A (BPA) on the mouse spermatogenesis in vivo. Male ICR mice were orally injected on a daily basis with low dose of BPA 20 mg/kg, high dose of BPA 200 mg/kg, or corn oil (vehicle control) for 7 days, and litter size and weights of body, testis, and cauda epididymis were measured. The level of serum testosterone and the expression of TGF- $\beta$$_1$ mRNA were also analyzed using RIA and RT-PCR, respectively. Also, morphological differences of testes after treatments were examined. Sperm concentration and level of serum testosterone showed a decreasing tendency detected as untreated >corn oil >low >high dose BPA treated mice, although there were no significant statistical differences. Interestingly, in mice treated with a high dose of BPA, partial disappearance of spermatozoa in seminiferous tubular lumen and the expression of TGF-$\beta$$_1$ mRNA were observed. Spermatogenesis was disrupted through TGF-$\beta$ system in the seminiferous tubules, resulting in no development of germ cells. Similarly, the litter size treated with a high dose of BPA was significantly different from that of untreated control group. In conclusion, these results that a high dose of BPA (200 mg/kg) acts as an endocrine disruptor during apermatogenesis in male mice md that there are BPA-specific lesions in the adult male reproductive tract might represent a permanently altered responsiveness to testosterone by BPA in the affected target tissue.