• Journal of Embryo Transfer
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• v.15 no.1
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• pp.95-102
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• 2000

Techniques for manipulation of spermatozoa and oocytes have been widely used for in vitro production(IVP) of Hanwoo. This study was conducted to examine the effects of theophylline and heparin on frozen-thawed Hanwoo sperm for enhancing the efficiency of IVP technique. Oocytes were inseminated with forzen bull semen treated with either theophylline or heparin for examining the effect of each substance on fertilization and subsequent development. More (P<0.05) oocytes formed pronucleus and develop to the morula and blastocyst stages after inseminated with sperm treated with heparin than after inseminated with sperm treated with theophylline. The pregnancy rate after embryo transfer was higher after heparin treatment than after theophylline treatment, but did not differ significantly. There was no significant difference of offspring delivery between two groups. In conculsion, theophylline and heparin can be used for enhancing the efficiency of IVP system for Hanwoo. Considering characteristics of these substance, theophylline may be useful in the artificial insemination system, which requires vigorous sperm motility. While, heparin supporting sperm viability in vitro can be effectively used for improving in vitro-fertilization system.

• Journal of Embryo Transfer
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• v.29 no.2
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• pp.163-169
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• 2014

The objective of this study was to evaluate the characteristics of Korean Native Cattle sperm frozen-thawed with L-cysteine and/or catalase. The semen from bulls was collected by the artificial vagina method, and Triladyl containing 20% egg-yolk and/or L-cysteine (L), catalase (C) and L-cysteine + catalase was added to the diluted semen for cryopreservation. The results showed that sperm viability was significantly higher in the L-cysteine + catalase ($69.49{\pm}3.16%$) group than in the control ($60.5{\pm}3.94%$) group (p<0.05). Acrosome damage was significantly lower in the L-cysteine ($17.12{\pm}1.08%$) group than in the control ($21.46{\pm}1.14%$), catalase ($20.54{\pm}0.76%$), and L-cysteine + catalase ($19.29{\pm}0.65%$) groups (p<0.05). In addition, the level of intact mitochondria in the spermatozoa was significantly higher in the L-cysteine ($58.65{\pm}1.39%$) group than in the control ($50.63{\pm}2.37%$) group (p<0.05). The hydrogen peroxide level in the frozen-thawed sperm was significantly lower in the L-cysteine ($3.74{\pm}1.66%$), catalase ($4.65{\pm}1.87%$), and L-cysteine + catalase ($8.11{\pm}2.15%$) groups than in the control ($13.22{\pm}1.6%$) group (p<0.05). The glutathione level was significantly higher in the L-cysteine ($1.33{\pm}0.03%$) group than in the control ($1.08{\pm}0.06%$), catalase ($1.05{\pm}0.02%$) and L-cysteine + catalase ($1.11{\pm}0.03%$) groups (p<0.05). In conclusion, L-cysteine and catalase could protect the membrane of Korean Native Cattle sperm from damage during sperm cryopreservation. Especially, L-cysteine was more effective for keeping acrosomes and mitochondria intactness during sperm cryopreservation.

• Korean Journal of Oriental Medicine
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• v.3 no.1
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• pp.85-104
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• 1997

As one of the chemical and experimental study on cancer treatment in Korea oriental medicine, we investigated the clinical characteristics on the treatment of cancer patients in oriental medical hospital. Furthermore, we carried out the cytotoxicity test of extracts of herbal medicine. The results are as follows: About Clinical Aspects of the Orient낄 Medical Cancer Treatment- (1) Clinical experience is that 420 Oriental medical hospitals were surveyed. Number of cancer patiens treated by one oriental medical doctor was under 10(about 60%)or from 10 to 20(about 30%). (2) Diagnostic Procedure Is that more than 80% of cancer patients were diagnosed by medical doctor of western hospital. (3) Therapeutic Method is that combination of oriental and western medical treatment was used in oriental medical hospital. (4) Relationship between cancer and constitutions is that to the relationship between cancer and constitutions, about 75% responded and most were considered to be Um. About Experimental Study- Toevaluate the direct cytotoxicity of hot water and methanol extracts of 34 oriental medical prescriptions on stomach cancer cell, SRB assay was carried out. (1) The $IC_{50}$ value of water extract of Samsoum was $19{\mu}g/ml$ and that of methanol extract was $15{\mu}g/ml$ so that we confirmed Samsoum has a strong direct cytotoxicity. (2) The $IC_{50}$ values of water extract of Coptidis Rhizoma, methanol extract of Caesalpiniae Lignum and Draba Semen were $10{\mu}g/ml$, $9{\mu}g/ml$ and $12.5{\mu}g/ml$ respectively, so that we confirmed Coptidis Rhizoma, Caesalpiniae Lignum and Drabs Semen have a strong direct cytototoxicity. (3) The $IC_{50}$ lalue of combinatory prescription of Rhei Rhizoma, Forsythie Fructus and Caesalpiniae Lignum was about $40{\mu}g/ml$.

• Korean Journal of Poultry Science
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• v.13 no.2
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• pp.197-208
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• 1986

This experiment was conducted to find out the effects of the six different feeding methods on the development of body weight, testis, comb and pituitary gland, and the sexual maturity of White Plymouth Rock cockerels. From hatching to 22 weeks of age, the weights of whole body, testis, comb and pituitary gland, and the histological changes of testis and the semen characteristics were checked every other week. The results obtained in this expeniment were as follows: 1. The growth rates of the self-feeding groups were faster than those of the limited feeding groups (70 percent of the self-feeding) by about 2 weeks. The weights of testis and comb showed the most marked increase at 20 weeks of age in the self-feeding groups and at 22 weeks of age in the limited feeding groups, respectively. 2. The weights of pituitary gland from hatching to 22 weeks of age at all observation weeks were not recognised significantly among the compared groups except 4, 14 and 16 weeks of age. 3. Correlations between week of age, body weight, testis, comb and pituitary gland, in the course of 22 weeks, were highly significant. 4. The diameters of lumina and tubules in the seminiferous tubules increased very slowly until 10 weeks of age. They showed the most marked increase at 12 weeks of age in the self-feeding groups and at 14 weeks of age in the limited feeding groups, and then continuously increased until 32 weeks of. age. 5. Primary spermatocytes appeared at first at 8 weeks in the all treatment groups, Secondary spermatocytes appeared at first at 10 weeks in the self-feeding groups and at 12 weeks in the limited feeding groups. At 14 weeks of age spermatids and spermatozoa were found at first in the self-feeding groups but spermatids were found in the limited feeding groups. 6. Age of the first ejaculation was between 14 and 16 weeks of age in the all treatment groups. The Average semen. volume and sperm concentration ranged from 0.1-0.2$m\ell$/ ejaculate and 5.6-9.8${\times}$10$\^$8/ sperm/$m\ell$ at the age of the first ejaculation but 0.30-0.35$m\ell$/ ejaculate and 22.4-42.7${\times}$10$\^$8/ sperm int at the 20 weeks of age in the all treatment groups.

• Reproductive and Developmental Biology
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• v.40 no.3
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• pp.27-31
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• 2016

The aim of this study was to evaluate effect of ${\alpha}$-linolenic acid (ALA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved in 20% egg yolk freezing extender containing ALA (0, 3, 5, and 10 ng/mL) with 0.05% ethanol. The frozen-boar spermatozoa were thawed at $37.5^{\circ}C$ for 45 sec in water-bath. The spermatozoa samples were evaluated the plasma membrane integrity, acrosome reaction, and mitochondrial integrity using flow cytometry. In results, population of live sperm with intact plasma membrane was significantly higher in control and 3 ng/mL ALA treatment group than ethanol group (p<0.05). In contract, dying sperms were higher in ethanol group than 3 ng/mL ALA treatment (p<0.05). Acrosomal membrane damage in all sperm population was reduced in 3 ng/mL ALA groups compared with ethanol treatment (p<0.05). However, acrosome damage in live sperm population was no significant difference among the all treatment groups. Mitochondrial integrity was not influenced by ALA treatments in both of live and all sperm population. In conclusion, this results show that supplement of ALA during the cryopreservation process could reduce the membrane damages including plasma and acrosomal membrane, whereas ALA did not influence to mitochondria in boar spermatozoa. Therefore, these results suggest that ALA can protect against the membrane damage derived cryo-stress, and cryopreservation efficiency of boar semen would be improved by use of ALA.

• Journal of Embryo Transfer
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• v.27 no.1
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• pp.45-50
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• 2012

The objective of this study is to estimate the effect of adding TES to LEY and FGE freezing extender for the sperm viability, acrosomal morphology and DNA fragmentation from miniature pig sperm, we evaluated sperm characteristics in TFGE, TLE and LEY with various thawing condition ($37^{\circ}C$ for 20 sec, 45 sec and $75^{\circ}C$ for 5 sec, respectively), and in different concentration of glycerol at 1%, 1.5%, 3%. The sperm viability and normal acrosome intact(NAI) in TFGE (Viability : $60.3{\pm}2.4$, NAI : $58.6{\pm}2.2%$), TLE ($61.3{\pm}2.4$, $62.2{\pm}2.2%$) extender significantly(p<0.05) increased than that in LEY ($50.2{\pm}2.4$, $54.5{\pm}2.2%$) extender thawed at $75^{\circ}C$ for 5 sec. According to the results from glycerol concentration, the viability and NAI of miniature pig sperm in 1.5% glycerol TLE ($66.1{\pm}3.2$, $66.2{\pm}1.0%$) was highest among the experimental groups. In accordance with this, DNA fragmentation rates was the lowest in TLE ($43.3{\pm}0.5%$) while that in LEY ($63.5{\pm}2.3%$) is the highest. Therefore, these results suggest that TLE extender method for freezing- thawing of miniature pig sperm increased the viability after thawing.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.9
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• pp.251-258
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• 2020

This study investigated the effect of Zardaverine supplementation in freezing extender, on kinetic characteristics of post-thawed boar sperm. Cryopreservation of boar sperm is an important technique of assisted reproductive technology and genetic resource banking. Although this technique is particularly useful, freeze-thaw cycles associated with sperm cryopreservation significantly reduce sperm quality. Semen from mature Duroc boars were collected and cryopreserved in freezing extenders (LEY) treated with varying concentrations of Zardaverine (0, 20, 50, 75, 100 𝜇M). The time-dependent kinetic characteristics of post-thawed spermatozoa were determined after thawing by applying computer-assisted sperm analysis (CASA). We observed that the motility immediately after thawing was significantly higher in 20 𝜇M stocks than in control (0 𝜇M) and the other treatments (p<0.05). Curvilinear velocity (VCL) in 0 𝜇M and 20 𝜇M stocks were significantly higher than the other treatment groups, except 75 𝜇M (p<0.05). Higher average path velocity (VAP) was obtained at 20 𝜇M as compared to 100 𝜇M, whereas amplitude of head lateral displacement (ALH) was significantly higher at 20 𝜇M than 50 𝜇M and 100 𝜇M (p<0.05). No differences were obtained for Straight-line velocity (VSL) and Linearity (LIN). In conclusion, our results indicate that Zardaverine improves the motility, VCL, VAP, and ALH of post-thawed boar sperm.

• Journal of Oriental Medical Thermology
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• v.4 no.1
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• pp.23-28
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• 2005

Purpose : To examine the relation between male infertility and cold hypersensitivity on lower abdomen. Methods : From 2004, 4 to 2005, 10, a total 60(infertile male 30, normal male on semen analysis 30) patients who visited the Oriental gynecological department of Conmaul Oriental Medicine hospital was selected, and their medical records were reviewed retrospectively. We compared the temperature of 3 points(CV17, CV12, CV4) of both group by DITI(Dorex, DITI-16UTI and DITI-Spectrum 9000 MB). Results and Conclusion : The general characteristics such as age, weight, height, BMI, amount and frequency of drinking and smoking of both group were not different statistically. The mean ${\Delta}T1$ between upper and lower abdomen in infertile group was higer than in normal group but they were not different statistically. The mean ${\Delta}T2$ between chest and lower abdomen was $0.46{\pm}0.41$ in infertile group, and $0.18{\pm}0.56$ in normal group, and it shows significant difference statistically. There was no significant relationship between sperm motility and ${\Delta}T1$, ${\Delta}T2$ in infertile group. In this study, we suggest that an inclination of cold hypersensitivity of lower abdomen of infertile male, but it is expressed when it is contrasted to chest of the body rather than upper abdomen.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.36-36
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• 2001

임신기의 자성 생쥐에 저 농도의 BPA의 투여가 태아의 체중, 태아수 및 성비에 미치는 영향을 검토하고, 태어난 자손의 번식기관무게, 정액성상, 조직검사 및 혈액의 혈구 성분분석등 번식효율과 생리기능에 미치는 영향을 검토하고자 실시하였다. 자성 생쥐에 corn oil(대조구), 0.05, 0.5 및 5.0mg BPA/kg b.w의 용량으로 1회 투여하고 투여 당일 날 웅성 생쥐와 교배 시켰으며, 3일 간격으로 5회 투여하였다. 임신기에 BPA의 투여가 웅성 자손의 체중에 미치는 결과는 각각 30.72, 30.80, 29.80 및 26.80g으로 5.0mg 투여구가 여타구에 비해 통계적으로 유의하게 낮은 체중을 나타냈으며(P<0.05), 정소의 무게는 BPA 투여구가 다소 높은 무게를 나타냈다. 정소상체, 정낭선 및 응고선의 무게는 투여구간에 커다란 차이가 없었다(P>0.05). BPA의 투여후 태어난 웅성 산자의 총정자수, 생존율, 유효정자수 및 기형율에 미치는 영향을 조사한 결과, 각 투여구간에 커다란 차이는 없었다. 또한 웅성 산자의 혈구화학치에 미치는 영향을 조사한 결과, RBC는 BPA 투여구가 각각 10.60, 10.96 및 10.35$\times$$10^{6}$㎣로서 대조구의 9.78$\times$$10^{6}$㎣보다 통계적으로 유의하게 높은 수치를 나타냈지만(P<0.05), WBC, Hb, HT, MCV, MCH, MCHC 및 PLT는 각 투여구간에 커다란 차이는 없었다. 임신기에 BPA의 투여가 자성 산자의 체중에 미치는 결과는 각각 25.56, 25.38, 25.83 및 23.86g으로 5.0mg 투여구가 다소 낮은 체중을 나타냈다. 자궁의 무게는 BPA 투여구가 각각 0.0915, 0.0904 및 0.1016g으로 대조구의 0.0801g에 비해 다소 높은 무게를 나타냈으며, 난소의 무게도 투여구간에 커다란 차이가 없었다. BPA의 투여가 자성 산자의 혈구화학치에 미치는 영향을 조사한 결과, WBC는 5.0mg 투여구가 여타구보다 다소 낮은 수치를 나타냈으며, PLT는 BPA 투여구가 대조구보다 다소 높은 수치를 나타냈지만 커다란 차이는 없었다 RBC, Hb, HT, MCV, MCH 및 MCHC는 각 투여구간에 차이가 없었다. 임신기에 BPA의 투여가 자손의 정소, 난소 및 자궁의 조직, 병리학에 미치는 영향을 조사한 결과, 정소와 난소의 조직에는 각 투여구간에 차이가 없었으며, 자궁에서는 0.5mg 투여구와 5.0mg 투여구에서 자궁내막층이 현저하게 증식된 것으로 나타났다.

• Reproductive and Developmental Biology
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• v.40 no.4
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• pp.33-37
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• 2016

This study was conducted to evaluate effect of ${\alpha}$-linolenic acid (ALA) and bovine serum albumin (BSA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved using freezing extender containing 3 ng/mL ALA and/or $20\;{\mu}g/mL$ BSA. Cryo-preserved boar sperms were thawed in $37^{\circ}C$ water-bath for 45 sec to analysis. Viability, acrosome reaction, and mitochondrial intact were analyzed using flow cytometry. In results, viability of frozen-thawed boar sperm was significantly higher in only ALA+BSA supplement group than control group (p<0.05), whereas there was no difference either in ALA or BSA supplement. However, acrosome reacted sperm in both of live and all sperm population were significantly decreased in all treatment groups than control (p<0.05). Interestingly, mitochondrial intact of boar sperm was enhanced in ALA and ALA+BSA groups compared with control (p<0.05). In this study, we showed that supplementation of ALA and BSA in freezing extender enhanced the sperm viability, mitochondrial intact and decrease acrosomal membrane damage. In conclusion, our findings suggest that quality of frozen-thawed sperm in mammalians could improve by using of ALA and BSA.