• Korean Journal of Animal Reproduction
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• v.14 no.4
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• pp.237-244
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• 1990

These studies were carried out to investigate the effects of the follicles size, hormone supplementation, semen types and capacitation methods on in vitro maturation and fertilization of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean Native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluid from the visible follicles of diameter 3~5mm. The follicular oocytes were cultured in TCM-199 medium containing hormones, FCS, ECS, BFF and MCC for 24~48hrs. in a incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 18~20hrs. with motile capacitated sperm in the TCF(Tyroide calcium-free) solution containing 200$\mu\textrm{g}$/ml of heparin. The results obtained in these experiments were summarized as follows : 1. The oocytes classified as "A, B, C, D and Degenerative" depending morphological integrity and those were 61.4%, 12.1%, 19.2%, 4.2% and 3.0% of the total oocytes recovered, respectively. The maturation and fertilization rate of the A, B, C class follicular oocytes, cultured in the TCM-199 medium supplemented with 10% FCS were 89.1%, 78.0%, 52.6% and 78.1%, 66.1%, 33.3%, respectively. 2. The average number of the follicular oocytes recovered from follicles size, 1~2mm, 3~5mm and above 5mm in dimeter were 67, 98 and 63, respectively. The maturation and fertilization rate of the follicular oocytes, cultured in the TCM-199 medium were 56.7%, 82.5%, 46.0% and 44.8%, 71.4%, 28.6%, respectively. 3. The maturation and fertilization rate of follicular oocytes, cultured in the TCM-199 medium supplemented with 5%, 10%, 15%, 20% FCS and hCG, HCG, $\beta$-estradiol were 76.0%~82.3% and 26.2%~70.0%, and those values were higher the supplementation of the hormone than the non-supplementation. 4. The fertilization and cleavage rate of the follicular oocytes, inseminated with spermatozoas of epididymis cauda, neat and frozen semen were 63.3%, 73.3%, 70.0% and 32.7%, 37.8%, 38.3, respectively. 5. The fertilization and cleavage rate of follicular oocytes, fertilized with capacitated spermatozoas by heparin, BFF and HIS methods were 70.0%, 53.8%, 34.2% and 38.3%, 23.1%, 17.1%, respectively. And the fertilization and cleavage rate were higher method of heparin than other methods.r methods.

• Journal of Life Science
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• v.26 no.9
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• pp.1063-1073
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• 2016

Male climacteric syndrome, andropause, or testosterone deficiency syndrome (TDS) is one of the new health issues in elderly men. It is a natural phenomenon that happens with age in men, which is clinically characterized by a decline in levels of serum testosterone resulting in a significantly decreasing of physical and mental activities. The aim of this study was to investigate the putative alleviative effects of dandelion extract on the symptoms of TDS by increasing serum testosterone levels and compare the efficacy between dandelion extract (DE) and fermented dandelion extract (FDE). After daily intake of DE and FDE for 4 weeks, serum testosterone levels, muscles of vastus lateralis, forced swimming time, total sperm counts, and motile sperm counts were significantly increased in older rats (22 weeks). Additionally, SHBG, epididymal fat pad, total serum cholesterol and triglyceride were significantly decreased in DE and FDE fed groups. However, PSA levels were not different among all groups. Furthermore, DE and FDE enhanced the expression of genes related to testosterone biosynthesis in TM3 Leydig cells. Overall, these positive effects on andropause were greater in FDE compared to DE. These results suggest the potential of FDE as a safe and efficacious natural material for recovering testosterone levels and reducing andropause symptoms.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.227-233
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• 2017

Pig has been known to be one of the most feasible animals as a bioreactor to produce pharmaceuticals in milk and as a mediator in xenotransplantation research. Previously, we generated transgenic pigs for both purposes, which were expressing Factor 8, vWF, hTPA, and hEPO in milk, along with expression of MCP at GalT gene locus ($GalT^{-MCP/-MCP}$) as well as expressing MCP at GalT gene loci with CD73 expression ($GalT^{-MCP/+}/CD73$). In this study, we performed comparative analyses of sperm parameters between wild type male (WT) pig and those transgenic males to examine the effects of transgenes integrated into the pigs on motility, morphology, viability, and acrosome integrity of the spermatozoa. Our results showed that the rates of actively motile spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 85.0%, 83.3%, 82.5%, 83.3%, 82.5%, 77.5%, and 78.7%, respectively. Whereas, the rates of morphologically normal spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 90.0%, 80.0%, 80.0%, 83.3%, 85.0%, 91.8%, and 80.8%, respectively. In addition, the viability in spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 93.9%, 82.4%, 89.9%, 83.9%, 87.4%, 92.8%, and 83.6%, respectively. The rates of spermatozoa with normal acrosome integrity in WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 98.1%, 98.6%, 98.6%, 98.7%, 98.1%, 99.5%, and 95.1%, respectively. There were no significant differences in motility, morphology, viability, and acrosome integrity of the spermatozoa among WT, Factor 8, vWF, hTPA, and hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs. These mean that neither random integration nor targeted integration of the transgene into chromosome of pig effect on characteristics of spermatozoa. Ultimately, the transgenic male pigs subjected in this study could apply to propagate their progenies for production of human therapeutic proteins and advancing the xenotransplantation research.

• Korean Journal of Poultry Science
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• v.41 no.1
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• pp.21-27
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• 2014

To preserve chicken genetic materials like cryopreserved spermatozoa, various kinds of freezing agents like glycerol, dimethylsuloxide, dimethylformamide or dimethylacetamide have been used for rooster semen preparation. Recently, the usage of N-methylacetamide (MA) for Ogye rooster semen preservation resulted in hatched chicken successfully. In this study, we investigated the effects of 7, 9 and 11% of MA on the viability, fertility and hatchability of frozen-thawed rooster semen using artificial insemination. The results of viability, fertility and hatchability in frozen semen with 7%, 9% or 11% MA were $35.16{\pm}6.12%$, $67.83{\pm}15.3%$ and $66.2{\pm}16.3%$ of motile sperm rate, 21.5%, 34.7% and 25% of fertility rate, and 100%, 89.5% and 87.5% of hatchability rate. The results of control group with frozen semen were 96.0% of fertility rate and 92.2% of hatchability rate. With these results, the concentration range of MA as a freezing agent of rooster semen could be 7~9% of media. The higher concentration of 9 % MA could decrease the fertility rate of thawed semen not the rate of hatchability rate. So the use of MA without affecting fertility rate would be a key point of freezing method of rooster semen for poultry genetic resource preservation.

• Korean journal of applied entomology
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• v.59 no.1
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• pp.25-35
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• 2020

The striped fruit fly, Zeugodacus scutellata, is endemic in Korea, but it has been regarded as one of the serious quarantine pests throughout the world. Sterile insect release technique (SIT) has been used to eradicate quarantine fruit flies. This study developed a technique to generate sterile males and applied SIT to control Z. scutellata. First of all, the reproductive systems of Z. scutellata adults were examined with fluorescent microscope. Polytrophic ovaries comprises of around 100 follicles with developing oocytes. Each follicle contains an oocyte with several nurse cells and are surrounded with follicular epithelium. Oocyte development began at 10 days after adult emergence (DAE) and formed chorionated oocytes after 20 DAE. On the other hand, male testes were well developed just after adult emergence. The vas deferens was filled with motile sperms. To generate sterile males, different doses (0~1,000 Gy) doses of electron beam were irradiated to 3~5 days old pupae of Z. scutellata. When male pupae were irradiated with electron beam at 200 Gy, they developed and mated with females without any significant difference compared to untreated males. Although the untreated females mated with the 200 Gy-irradiated males laid eggs, no eggs did not hatch. The 200 Gy-irradiated males were then applied to untreated male and female flies in a density ratio of 1:9 (untreated males : treated males). The laid eggs suffered significant infertility. These results suggest that electron beam-irradiated pupae at 200 Gy resulted in male sterility and the resulting males would be applied to SIT.

• Journal of the Korean Applied Science and Technology
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• v.30 no.1
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• pp.116-126
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• 2013

Glycol ethers are a group of solvents based on alkyl ethers of ethylene glycol commonly used in paints. These solvents typically have a higher boiling point, together with the favorable solvent properties of lower-molecular weight ethers and alcohols. The word "Glycol ethers" was registered as a United States trademark by Union Carbide Corp. Typically, glycol ethers are found in pharmaceuticals, sunscreens, cosmetics, inks, dyes and water based paints. On the other hand, glycol ethers are used in degreasers, cleaners, aerosol paints and adhesives. Most glycol ethers are relatively water soluble, biodegradable and only a few are considered toxic. Therefore, they are unlikely to pose an adverse risk to the environment. Recent study suggests that occupational exposure to glycol ethers is related to low motile sperm count in men, but the finding has been disputed by others. In this study, skin permeation of 3 types glycol ethers were studied in vitro using matrix such as solvent and detergent. The absorption of glycol ethers[methyl glycol ethers(MC), ethyl glycol ethers(EC) and butyl glycol ethers(BC)] has been measured in vitro through rat skin. Epidermal membranes were set up in Franz diffusion cells and their permeability to PBS measured to establish the integrity of the skin before the glycol ethers were applied to the epidermal surface. Absorption rates for each glycol ethers were determined and permeability assessment made to quantify any irreversible alterations in barrier function due to contact with the esters. Types of glycol ethers in vitro experimental results on MC> EC> BC quickly appeared in the following order: skin permeation was beneficial to the skin permeation small molecular weight, the difference in chemical structure, such as hydrophilic, because with the partition coefficient and solubility mechanisms and passive diffusion to increase the speed at which transmission is considered.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.255-262
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• 2006

The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.7
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• pp.790-800
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• 2017

As men get older, total testosterone levels decline gradually, and concentrations of free and bioavailable testosterone decline sharply with each decade beyond their 30s. Andropause or testosterone deficiency syndrome (TDS) is defined as a decrease in sexual satisfaction or decline in general well-being accompanied by low levels of testosterone in older men. This male climacteric is characterized by nervousness, reduced potency, decreased libido, irritability, fatigue, depression, memory problems, sleep disturbances, and hot flushes. Cirsium japonicum (CJ) is used as a traditional medicine for hemorrhage, blood congestion, and inflammation in Korea. However, there is no report on the efficacy of CJ treatment for TDS. In this study, we observed the mitigating effect of CJ extract (CE) and fermented CJ extract (FCE) on symptoms of TDS. In elderly male rats, total and testosterone levels, hind limbs muscles, forced swimming time, and total and motile sperm counts significantly increased after daily intake of CE and FCE for 6 weeks. In contrast, sex hormone binding globulin, retroperitoneal fat, total serum cholesterol, and triglyceride levels were significantly reduced in CE and FCE groups. However, there was no difference in prostate specific antigen, aspartate aminotransferase, and alanine aminotransferase levels among all groups, which means CE and FCE did not have putative adverse effects. In a cell experiment, we also observed that CE and FCE enhanced expression of genes related to testosterone biosynthesis but reduced genes involved in testosterone conversion. On the whole, these positive effects on TDS were greater in FCE compared to CE. Thus, these results suggest the potential of FCE as a promising natural product for recovering testosterone levels and alleviating TDS.