• Clinical and Experimental Reproductive Medicine
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• v.32 no.2
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• pp.149-154
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• 2005

연구목적: 정자의 첨체상태는 수정능과 상관관계가 있다. 본 연구는 사람 정자의 동결보존 시 Fertilization promoting peptide (FPP) 처리가 첨체 유지에 효과가 있는지를 알아보고자 실시하였다. 연구재료 및 방법: 사람 정자는 정액검사를 의뢰한 시료를 사용하였으며, 적정농도를 조사하기 위하여 25, 50, 100 nM FPP를 신선정자에 처리한 뒤 시간별로 첨체의 변화를 조사하였다. 또한 적정화된 50 nM FPP를 정자의 동결-융해 시에 처리한 뒤 첨체 변화를 조사하였다. 첨체 변화는 FITC - pisum sativum lectin (PSA) 염색방법을 이용하여 조사하였다. 결 과: FPP 농도 변화와 처리시간에 따른 사람 정자의 첨체 변화를 조사하였던 바, 50 nM FPP 처리군에서 대조군보다 높은 온전한 첨체비율을 얻을 수 있었다. 정자의 동결-융해 시, 동결액과 융해액에 50 nM FPP 첨가가 온전한 첨체를 유지하는 비율을 조사하였던 바, 신선 정자의 결과보다는 유의하게 낮지만 무 처리군보다 유의적으로 높은 온전한 첨체를 얻을 수 있는 것을 알 수 있었다. 또한 동결액에만 또는 융해액에만 50 nM FPP 처리를 하더라도 무 처리군보다 유의하게 높은 온전한 첨체 비율을 획득할 수 있음을 알 수 있었다 (p<0.001). 결 론: 사람 정자의 동결보존 시 50 nM FPP 첨가는 자발적으로 발생하는 첨체반응을 억제하고, 온전한 첨체를 유지할 수 있어 수정능 보유에 기여할 수 있을 것으로 사료된다.

• Journal of Korean Ophthalmic Optics Society
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• v.7 no.2
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• pp.189-195
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• 2002

The aim of this study was to investigate the action of benzalkonium chloride (BAC) used as a preservative in most ophthalmic topical solutions, on human corneal epithelial (HCE) cells in vitro. HCE cell line was exposed to BAC solutions at various concentrations (0.01%~0.0001%) for 15 minutes followed by 24 hours of cell recovery. Cell viability was assessed using MTT assay and chromatin condensation with a Hoechst 33342 test. The expression of membrane protein Fas and Fas ligand was examined by western blot and immunocytochemistry, and DNA fragmentation was studied by agarose gel electrophoresis. A significant decrease of membrane integrity with chromatin condensation was observed with BAC tested at concentrations of 0.005% and higher. BAC was cytotoxic preservatives in this study. An apoptotic mechanism appeared to be present at low concentrations of BAC, whereas a necrotic process appeared at higher concentrations. A functional Fas-mediated apoptotic pathway is present in cultured HCE cells and can be activated by upregulation of Fas expression with BAC.

• Journal of Aquaculture
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• v.20 no.3
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• pp.173-177
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• 2007

An experiment was performed to obtain cryopreservation techniques of starry flounder (Platichthys stellatus) sperm. Milt obtained from 24 males were cryopreserved using two diluents, artificial seminal plasma (ASP) and Stein's solution (SS) with three cryoprotectants, dimethyl sulfoxide (DMSO), methanol, and glycerol concentrated from 5% to 20%. Post-thaw sperm activity (motility and/or speed) revealed the highest in 10% DMSO and 15% methanol in ASP and SS as diluent. Motility and speed of cryopreserved sperm were decreased according to increase of glycerol concentration. To conclude, DMSO was a better cryoprotectant than methanol or glycerol for cryopreservation of starry flounder sperm. Glycerol was incongruent cryoprotectant because of toxic to starry flounder sperm. Most cryopreserved spermatozoa without cryoprotectant showed the enlarged head with granulated chromatin and ruptured plasma membrane by freezing and thawing injuries compared with unfrozen normal spermatozoa.

• Proceedings of the KTCVS Conference
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• 1995.10a
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• pp.119-119
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• 1995

• Proceedings of the Korean Society of Developmental Biology Conference
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• 1998.07a
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• pp.34-35
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• 1998

• 보존과학연구
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• s.32
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• pp.25-35
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• 2011

Wood vinegar and Asarum sieboldii Miquel were selected as candidate materials showed antimicrobial activity and insect repellent activity. These natural pesticides have its own color and these could cause color difference on fabric materials. In the present study, we investigated the color difference of undyed and dyed fabrics to evaluate negative effect of wood vinegar and A. sieboldii Miquel. Undyed and dyed fabrics were exposed to natural pesticides of various concentrations for six months in relative humidity 70% and temperature $28^{\circ}C$. After exposure of pesticides, color difference (${\Delta}E*$) were investigated at two weeks intervals for six months. As a results, dyed cotton, silk and undyed silk fabrics exposed wood vinegar were not nearly changed in their colors, but color of only undyed cotton fabric was clearly changed by wood vinegar. Especially color difference by wood vinegar on undyed cotton fabric was most distinct as the concentration increased. On the other hand, all of fabrics exposed A. sieboldii Miquel were not nearly changed in their colors for six months. Therefore, this study first suggests that wood vinegar and A. sieboldii Miquel as natural insecticides could be used to conserve for textile cultural properties from insects and microorganism, but wood vinegar couldn't use the high concentration on undyed cotton fabric.

• Clinical and Experimental Reproductive Medicine
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• v.35 no.3
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• pp.203-212
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• 2008

Objective: During cryopreservation process, cold shock and cryo-injury affect the fertilizing capacity of the sperm by damaging cell membranes with loss of functional integrity. A longstanding concept for preventing the cryo-damage is to stabilize the plasma membrane by incorporating cholesterol. This study was to determine the effects of cholesterol in freezing media on the motility and functional integrity of human sperm after cryopreservation. Methods: Control group (non-cholesterol treated) and different concentrations of cholesterol-treated sperm (14 healthy males) were frozen and thawed. After freezing and thawing of sperm, the quality of sperm was evaluated by sperm analysis, acrosome reaction test and sperm chromatin structure assay. Results: When human sperm were incubated in sperm freezing medium (SFM) containing $0.5{\mu}g$ cholesterol and then freezing/thawing, the motility of sperm have significantly improved compared to those untreated cholesterol ($33.46{\pm}1.48%$ vs. $30.10{\pm}1.07%$, p<0.05). The rate of calcium ionophore-induced acrosome reactions in post-thawed sperm was significantly higher than that ($53.60{\pm}1.60%$ vs. $47.40{\pm}1.86%$, p<0.05) in SFM containing cholesterol. Sperm chromatin structure assay revealed that DNA damage to the sperm in the cholesterol-treated group was lower than that of non-treated group. Conclusion: These results suggest that increased cholesterol content of sperm plasma membrane by supplementation of cholesterol in SFM improves sperm motility, capacitation status, and DNA integrity. Therefore, addition of cholesterol into SFM could be a useful for protecting human sperm from cold shock and cryo-injury during cryopreservation.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.195-201
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• 2003

This study was carried out to establish most suitable freezing condition, to evaluate the different condition of freezing and thawing rates on the viability and motility of frozen canine spermatozoa. The ejaculated semen was added to obtained 200∼400 ${\times}$ 10$^{6}$ /$m\ell$ with extender I and was cooled to 4$^{\circ}C$ over 30, 60 and 120 minutes. And then, semen was diluted with extender II containing 4, 6 and 8％(v/v) glycerol for 60 min, respectively and packaged in 0.5$m\ell$ straw, equilibrated far 30, 60 and 120 min at 4$^{\circ}C$ and cryopreserved in liquid nitrogen vapor at different distance(3, 5, 7 and 9 cm, respectively), plunged into nitrogen tank. Samples were thawed by placing straws into 27, 37, 47, 57$^{\circ}C$ water bath for 120, 20 and 12 sec, respectively. The results were as follows; 1. The survival and motility rate immediately post-thawing was significantly higher in samples frozen in 4％ glycerol than 6 or 8％ glycerol(P< 0.05). 2. According to equilibration time at 4$^{\circ}C$, the survival and motility rate immediately post-thawing was significantly higher in samples frozen after 60 min equilibration than 30 or 120 min equilibration(P<0.05). 3. Freezing in distance of 5 cm from liquid nitrogen yield better survival and motility rate than the others(3, 7 or 9 cm)(P<0.05). 4. The effect of thawing rate on sperm survival were higher when the thawing was done at 37$^{\circ}C$ for 120 sec(P<0.05).

• Parasites, Hosts and Diseases
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• v.32 no.1
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• pp.57-60
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• 1994

In order to search the fate of Thelohonellus kitauei spores the extrusion rates of the polar filaments were monitored in uipo chronologically. Preserved spores suspended with various solutions at $-70^{\circ}C$ showed almost the same vigorous pattern as early freezing stages up to 1,750 days after initial preservation. It revealed that the vlabllltles of some spores suspended with 0.45% and 0.9% NaCl solutions and distilled water at $5^{\circ}C$ continued for 1,628 days, 1,614 days and 1,721 days, respectively. And, the life spans of some spores in the previous solutions added with antibiotics at $5^{\circ}C$ were 1,628 days, 1,614 days and 1,714 days, respectively. Key words: TheLohanellus kitnuei, spore, extrusion rate of polar filament, life span

• Korean Journal of Food Science and Technology
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• v.34 no.4
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• pp.652-660
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• 2002

To develop natural food preservatives for extending the shelf-life of jeotkal (salted and fermented seafood), antimicrobial substances were extracted from 32 types of medicinal herbs and edible plants using 95% ethanol. Among the extracts, Glycyrrhizae radix, Curcumae domestica, Galla rhois, and Resina pini showed relatively high inhibitory effects on the growth of the microorganisms isolated from the deteriorated jeotkal. We selected and tested the extract from Recina pini as a natural jeotkal preservative. This ethanol extract was purified partially by adding equal quantity of water, through which 77% of insoluble materials were removed as impurities. In manufacturing modified jeotkal using squid, sucrose and starch syrup were substituted with sorbitol, $glucono-{\delta}-lactone$ was added instead of vitamin C and lactic acid, and sterilized hot pepper was used instead of natural one. The shelf-life of modified jeotkal was prolonged by 4 days compared with the control jeotkal when stored at $20^{\circ}C$, while that of modified jeotkal containing 1.0% partially purified Recina pini extract was prolonged by 6 days compared to the control. The same tests were conducted for the changran (stomach and intestine of Alaska pollack) jeotkal preservation. The shelf-life of the control jeotkal was 24 days, whereas the modified jeotkal and the Resina pini extract-containing modified jeotkal maintained their qualities without changes in microbial and chemical characteristics for 90 days at $20^{\circ}C$ storage.