• Title/Summary/Keyword: Semen preservation

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Effect of Methyl-Beta-Cyclodextrin (MBCD) on Cryopreservation of Bull Spermatozoa (소 정액의 동결 보존시 Methyl-Beta-Cyclodextrin (MBCD)의 영향)

  • Lee, Kyung-Jin;Seo, Gi-Beom;Lee, Yong-Seung;Yoo, Han-Jun;Cheong, Hee-Tae;Lee, Seung-Hwan;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.295-300
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    • 2011
  • The objective of this study was to evaluate efficiency of methyl-beta-cyclodextrin (MBCD) in the sperm preservation of bull. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 1, 5, 10 and 20 mM MBCD before freezing process. Analysis of viability in frozen-thawed sperm was estimated by SYBR14/PI double stain, hypoosmotic swelling test(HOST) and acrosome damage with FITC-PNA, and mitochondria activation with Rhodamin123 by flow-cytometry. The sperm viability was significantly in 0 mM and 5 mM concentrations of MBCD than other groups (p<0.05). However, the HOST was significantly lower at 20 mM concentration of MBCD than other concentrations (p<0.05). In addition, acrosome damage and mitochondria activation rates were significantly lower at 20 mM concentration of MBCD than other groups (p<0.05). In conclusion, the viability of sperm was not significantly different among concentrations of MBCD 0, 5 and 10 mM, but MBCD 20 mM was significantly lower than other groups. In addition, as concentrations of MBCD was high, HOST, acrosome damage and mitochondria activation rates had a negative effect in bull sperm.

Enhancing liquid-chilled storage and cryopreservation capacities of ram spermatozoa by supplementing the diluent with different additives

  • Rateb, Sherif A.;Khalifa, Marwa A.;El-Hamid, Ibrahim S. Abd;Shedeed, Hesham A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.7
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    • pp.1068-1076
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    • 2020
  • Objective: In the present study, we determined efficiency of incorporating caffeine, melatonin or omega-3 polyunsaturated fatty acid in the diluent on mitigating consequences of (a) liquid chilled- and (b) cryo-storage of ram spermatozoa. Methods: In the first experiment, ejaculates (n = 30) were collected from 5 adult rams and were pooled, diluted (1:10) with Tris-citric acid (base diluent) and were split into 4 aliquots assigned for: control (untreated), caffeine (0.1 mM), melatonin (0.3 mM) or omega-3 fatty acids (0.3 mM) (T0). The diluted specimens were stored at 4℃ for 48 h, during which sperm physical and cytological properties were evaluated along with oxidative stress indices (T24, T48). In the second experiment, 15 ejaculates (3 per male) were pooled, diluted with glycerolized base diluent (4% glycerol, v/v) and were split corresponding to the same previous treatment groups before being processed for cryopreservation. Post-thaw physical and kinematic sperm properties were assessed by a computer-assisted sperm analysis system. Results: The results clarified superiority of both melatonin and omega-3 supplementation on maintaining (p<0.05) sperm properties, while reducing (p<0.05) lipid peroxidase reaction and enzymatic activities of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase in preservation medium, compared to caffeine either during liquid-chilled storage or cryopreservation of spermatozoa. Conclusion: Melatonin and omega-3 are regarded efficient alternatives to caffeine when processing ram spermatozoa for application of artificial insemination or in vitro fertilization.

Successful onco-testicular sperm extraction from a testicular cancer patient with a single testis and azoospermia

  • Kuroda, Shinnosuke;Kondo, Takuya;Mori, Kohei;Yasuda, Kengo;Asai, Takuo;Sanjo, Hiroyuki;Yakanaka, Hiroyuki;Takeshima, Teppei;Kawahara, Takashi;Kato, Yoshitake;Miyoshi, Yasuhide;Uemura, Hiroji;Iwasaki, Akira;Yumura, Yasushi
    • Clinical and Experimental Reproductive Medicine
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    • v.45 no.1
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    • pp.44-47
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    • 2018
  • Onco-testicular sperm extraction is used to preserve fertility in patients with bilateral testicular tumors and azoospermia. We report the case of a testicular tumor in the solitary testis of a patient who had previously undergone successful contralateral orchiectomy and whose sperm was preserved by onco-testicular sperm extraction. A 35-year-old patient presented with swelling of his right scrotum that had lasted for 1 month. His medical history included a contralateral orchiectomy during childhood. Ultrasonography revealed a mosaic echoic area in his scrotum, suggesting a testicular tumor. The lesion was palpated within the normal testicular tissue along its edge and semen analysis showed azoospermia. Radical inguinal orchiectomy and onco-testicular sperm extraction were performed simultaneously. Motile spermatozoa were extracted from normal seminiferous tubules under microscopy and were frozen. Eventual intracytoplasmic sperm injection using the frozen spermatozoa is planned. Onco-testicular sperm extraction is an important fertility preservation method in patients with bilateral testicular tumors or a history of a previous contralateral orchiectomy.

A Literature study on the Sasang Constitutional Tongue Diagnosis based on Experience Prescriptions(經驗方) in the Donguisusebowon (Longevity and Life Preservation in Oriental Medicine) (동의수세보원(東醫壽世保元) 경험방(經驗方)에 근거한 사상체질별 설진(舌診)의 문헌연구)

  • Lee, Jae-Chul;Kim, Jong-Yeol;Ryu, Hyun-Hee;Kim, Keun-Ho
    • Journal of Sasang Constitutional Medicine
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    • v.22 no.1
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    • pp.1-8
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    • 2010
  • 1. Objectives: This study was performed to research the relationship between Sasang Constitution and characteristics of tongue diagnosis based on Donguisusebowon's Experience prescriptions(經驗方). 2. Methods: We searched tongue diagnosis factors in Donguisusebowon and classified this. Based on Donguisusebowon's Experience prescriptions(經驗方), we find tongue diagnosis factors in 6 contemporary Oriental medicine prescription books. 3. Results: Most So-eumin has white thin tongue coating and clear redish tongue color. Yellow tongue coatings are found in symptoms for Yinjinho-tang(Yinchenhao-tang), Soseungki-tang(Xiaochengqi-tang), Daeseungki-tang(Dachengqi-tang). Shortened Tongue is found in So-eumin's experience prescription and So-yangin's treat histroy. It didn't shown that So-eumin's Semen Crotonis included 6 prescritions' factors of tongue diagnosis. So-yangin's white tongue coating is found in Hyungbangpaedok-san(Jingfangbaidu-san), Sipjo-tang(Shizao-tang), Sosiho-tang(Xiaocihu-tang), Shinki-hwan(Shenqi-wan). Most of other So-yangin's tongue coatings are Yellowish and tongue color is deep red. Tae-eumin have thin white tongue coating at the Mahuang-tang(Mahuang-tang). Red tongue color and drying tongue are found at Jowiseungki-tang(Tiaoweichengqi-tang)Daesiho-tang(Dacihu-tang)Saengmaek-san(Shengmai-san) 4. Conclusions: Following So-eumin's Experience prescriptions, So-eumin's tongue color is clear red and tongue coatings are thin white and greasy. Following So-yangin's Experience prescriptions, So-yangin's tongue color is deep red and tongue coatings are yellowish and scarce except Exterior cold disease. And So-yangin has specific symptom of irritability, oral sores. Tae-eumin's tongue has two sides of white thin tongue coatings and Red and drying tongue.

${\alpha}$-Glucosidase Inhibitory Substances Exploration Isolated from the Herb Extract (생약재 추출물로부터 분리한 ${\alpha}$-Glucosidase 저해활성물질 탐색)

  • Choi, Gil-Yong;Han, Gab-Jo;Ha, Sang-Chul
    • Food Science and Preservation
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    • v.18 no.4
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    • pp.620-625
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    • 2011
  • This is a part of the study on the food materials that are effective for diabetes treatment and for use in the development of functional bread products. In this study, various commercially available Oriental medicines with the intestinal absorption enzyme called ${\alpha}$-glucosidase, which is known to be effective for diabetes treatment, were explored. According to the research results on the water and methanol in 200 kinds of Oriental medicines, which were separated by layer to investigate the inhibitory activity of ${\alpha}$-glucosidase, Astragalus membranaceus (70.9%) in the water layer and Pericaeta communissima (72.9%) in the MeOH layer showed a strong inhibitory effect of over 70%. Myristica fragrans (69%), Morus alba (66.9%), Schisandra chinensis (65%), Panax notoginsens (63.9%), Anthriscus sylvestris (62.9%), Asparagus cochinchinensis (62.1%), Erycibe obtusifolia (60.9%), Polygonum cuspidatum (60.7%), Atractylodes lancea (60.2%), and Perilla frutescens (60.2%) in the water layer, and Codonopsis pilosula (67.8%), Prunus persica batsch (67.6%), Sinomenium acutum (63.5%), and Malvae semen (61.6%) in the MeOH layer, showed a more than 60% inhibitory effect. Thirty one species, including Polygonatum sibiricum (59.8%), Medicata fementata (59.7%), Alisma canaliculatum (59.5%), Coix lacryma-jobi (59.2%), Asiasarum sieboldi (59.0%), and Bupleurum falcatum (53.0%), in the water layer, and 10 species [Quisqualis indica (58.8%), Lycium chinense (58.3%), Trichosanthes kirilowii (58.0%), Thuja orientalis (55.9%), Bombyx mori (55.6%), Gallus domesticus (55.4%), Aralia continentalis (55.3%), Cibotium barometz (52.7%), Euphorbia pekinensis (52.7%), and Dolichos lablab (52.5%)] in the MeOHlayer, showed a more than 50% inhibitory effect. Therefore, such materials are expected to be the basic materials that will be used for the development of functional materials for diabetes treatment.

The Study on the History of Pugation therapy From -'Treatise on Febrile Diseases' to 'Longevity and Life Presservation In Oriental Medicine'- (하법(下法)의 발전 과정에 대한 연구(硏究) -상한론(傷寒論)에서 사상의학(四象醫學) 까지-)

  • Choi, Yei-Kwen;Kim, Kyung-Yo
    • The Journal of Internal Korean Medicine
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    • v.19 no.1
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    • pp.524-552
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    • 1998
  • Purgation therapy has played an important role as a influential remedy from the begining of the Chinese medicine. Especially purgation therapy is raised as the effective remedy on the acute infectious disease in the book of 'Treatise on Febrile Disease'. But It was inclined to cold-nature and available only in the excess syndrome. Nevertheless it is evident that the book has showed an example of this therapy. During the middle age, purgation therapy is classified into several subtype; hydrogogue therapy, laxation with lubricants, purgation with cold-natured drugs and purgation with warm-natured drugs. Comparing with the ancient times, it must be a progression. It was investigated earnestly by a school leaded by Zhang Congzheng. They were not restricted to several diseases, but applied it to the wide range of diseases. They thought as following. 'One is ill from pathogenic factor so that you should eliminate it from the human body'. Hence, they frequently used three major remedies such as diaphoresis, emesis and purgation. In this process, purgation therapy had showed eye-opening progress. But opposition to it was not little. Li Gao was a representative man on the opposite side. He expressed a critical opinion and placed great importance on the genuine energy, the natural healing force. Under his influence, a large number of doctors evaded purgation and put it under taboo. On account of these trend, purgation therapy had took a backward step and retrograded. Therefore cathartics such as Rhei Radix et Rhizoma, Rharbitidis Semen, cold drugs such as Gypsum Fibrosum, etc. had been excluded for preservation of the genuine energy, and came about an obnoxious custom to value only 'tonity deficiency', or 'warm and tonify'. As it had came into fashion to approach most disease from the point of view, purgation therapy was merely fall into a remedy of constipation. After the eighteenth century purgation therapy encountered the new period of rivival. It was introduced by them who strived for the study of Epidemics to the new current of thought, so called '增水行舟'. It was because 온병 was apt to dissipate one's Yin fluid. Therefore purgation therapy of this period was characterized by establishing nourishment Yin and body fluid with or without use of timely purgation of accumulation of heat. From the time of Zhang Congzheng, it was accomplished by Lee Je-ma to the most epoch-making change. He caused an improvement in the use of purgation therapy by regarding innate constitutional contradiction as importance than representing clinical symptoms. He warned that existing remedies that depend only upon symptoms and signs, not upon individual characteristics including constitutional features didn't bring round to but kill them. And he understood all the pathologic processes in his constitutional theory, investigated specific drugs on four constitution, made indications of each prescriptions clear. For giving to differentiation of constition before differentiation of syndrom, his new slant on the pathologic phenomena overcome the limitations of 변증시치, and revaluate purgation therapy from remedy impaire the genuine energy to that restore it by recover the balance between the internal organ. It is the product of him to fundamentally upset the cause to be in disregard of purgation therapy.

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Cryopreservation of Primordial Germ Cells(PGCs) from Korean Native Chicken(Ogye) Embryos using Commercial Cryoprotectants (상업용 동결보호제를 이용한 한국재래닭(오계) 원시생식세포의 동결 보존)

  • Kim, Hyun;Kim, Dong Hun;Han, Jae Yong;Do, Yoon Jung;Kim, Jae Hwan;Kim, Young Sin;Seong, Hwan Hoo;Ko, Yeoung Gyu;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.163-169
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    • 2013
  • Cryopreservation of poultry semen has been reported, but preservation of female genetic material has not been possible because of the unique anatomical and physiological characteristics of the avian egg. Thus an alternative strategy for conservation of oviparous species of animals must be developed. Recent technological developments for producing germline chimeras by the transfer of primordial germ cells (PGCs) into recipient embryos has enabled the conservation and retrieval of chicken genetic resources in their complete form. In the present study, fertilized eggs were incubated for about 5.5 days to obtain embryos at stage 28. The whole embryo was collected from the germinal gonad using a fine glass micro pipette under a microscope. The PGCs were then purified using MACS method. Two commercially available cryoprotectants (A and B) were used to preserve the PGCs, and EG were used as a control. The average recovery rate of PGCs after thawing was 35.5% and 60.5% with the A and B treatments, respectively. There was no significant difference between B treatments and control, which showed an average recovery rate of 52.8%. However, the recovery rate obtained using A cryoprotectant (35.5%) was significantly lower than using treatment control and B. The average viability of the PGCs after thawing were 77.9% and 77.4% for cryoprotectants A and B, respectively, and the control were was 81.6%. There was no statistically significant difference between the two treatments and control. It was concluded that all of the available cryoprotectants examined in this study could be used for preservation of PGCs from embryos. Further experiments to produce germline chimera from PGCs preserved using this techniques are strongly recommended.