• Clinical and Experimental Reproductive Medicine
• /
• 제16권1호
• /
• pp.9-14
• /
• 1989

For the cryopreservation of human embryos this study was accomplished as a preliminary experiment. The purpose of this study is to obtain optimal cryoprotectant, addition and dilution method of cryoprotectant and cooling rate for raising survival of frozon and thawed 2-cell mouse embryos. Seeding was done at $-7^{\circ}C$ and the straw contained embryos was plunged at $-30^{\circ}C$ when the slow cooling was ended. Embryos those developed normally to blastocyst after in vitro culture for over 96 hours were regarded as survival ones. The survival was the rate of number of survival embryos against the recovered embryos. The results are followed : 1. The survivals were 6.3, 71.2 and 67.4% respectively, when Glycerol, DMSO and 1,2-Propanediol were used as cryoprotectant. 2. When sucrose was added in freezing solution, the survival was 69.0%. That was higher than the survival of embryos frozen without sucrose in freezing solution. The difference was not significant. 3. Addition and dilution of cryoprotectant by 4 stepwise raised the survival than by direct, but that was not significant. 4. When embryos were frozen by -0.3, -0.5 and $-1^{\circ}C/min$ before plunged into $LN_2$, the survivals were 67.9, 78.0 and 37.0% respectively. The differnce was significant.

• Reproductive and Developmental Biology
• /
• 제28권3호
• /
• pp.187-190
• /
• 2004

PKC는 그들의 cofactor-requirments에 따라 cPKC, nPKC 그리고 aPKC, 3그룹으로 나어진다. 마우스 난 성숙과정에 있어서 cPKC 및 nPKC의 activators인 PMA의 영향에 대한 많은 결과가 보고되었다. 그러나 각각의 그룹에 대한 차별화된 영향에 대하여는 밝혀져 있지 않다. Mezerein의 analog인 thymeleatoxin은 cPKC의 특이적인 activator로 보고되어져 있다. 본 연구에서는 specific cPKC activator인 thymeleatoxin의 마우스 난 성숙과정에의 영향을 제1감수분열 재개 능(germinal vesicle break down, GVBD)과 제1 극체 형성 능(1st polar body extrusion)을 조사하여 cPKC및 nPKC activator인 PMA와 비교 검토하였다. 그 결과 GVBD IC50는 thymeleatoxin에서 ～400nM, PMA에서는 ～50nM이었으며, 제1극체 방출의 IC50는 thymeleatoxin에서 ～200nM, PMA에서는 ～20nM이었다. 이들 결과는 Thymeleatoxin의 GVBD나 1st polar body extrusion 저해효과가 PMA에 비하여 1/8～1/10인 것으로 나타났다. 이들 결과는 GVBD나 제1극체 형성을 포함하는 난 성숙과정에서 cPKC보다 상대적으로 nPKC의 관여가 깊음을 보여 준다.

• 대한한의학방제학회지
• /
• 제5권1호
• /
• pp.169-178
• /
• 1997

동의학(東醫學)에서는 질병의 발생에 대하여 정기(正氣)의 강약(强弱)에 그 관건(關鍵)이 있다하였다. 그리하여 모든 질병의 치료에 있어서도 정기(正氣)의 회복에 중점을 두는 부정법(扶正法)이나 거사법(祛邪法)을 사용하면서도 정기(正氣)에 손상(損傷)을 입히지 않도록 많은 관심을 기우리고 있다. 암(癌)의 치료 또한 정기(正氣)가 얼마만큼 보충되는가에 따라 항병능력(抗病能力)이 항진(亢進)된다할 수 있을 것이다. 현재 국내외적(國內外的)으로 악성종양(惡性腫瘍)을 치료하기 위하여 다방면으로 연구를 하고 있으며, 또한 실제로 사용하고 있는 치료방법들로는 수술요법(手術療法) 방사선요법(放射線療法) 면역료법(免疫療法) 화학요법(化學療法)등이 있지만 아직까지는 미흡한 상태라 할 수 있을 것이다. 현재 일반적으로 항암제(抗癌劑)를 이용한 화학요법(化學療法) 등이 사용되고 있고, 면역료법(免疫療法)을 위한 많은 방법들이 연구되고 있어 본 저자(著者)들은 천문동(天門冬)이 면역세포(免疫細胞)의 증식(增殖)을 촉진시켜주면서 항암작용(抗癌作用)이 있을 것으로 사료(思料)되는 바 피부암세포(皮膚癌細胞)인 A431 cell line과 골수암세포(骨髓癌細胞)인 KHOS-NP cell line에 천문동(天門冬)을 투여(投與)하여 암세포(癌細胞)의 증식(增殖)을 살펴보고, 면역세포(免疫細胞)인 T cell과 B cell의 증식(增殖)을 살펴보았다. 또한 복강내(腹腔內) macrophage에서 분비되는 NO의 양(量)을 In vitro와 In vivo실험(實驗)을 통하여 살펴보았고, T cell의 apoptosis 및 subpopulation의 양(量)을 관찰하였다. 이 결과(結果) 천문동(天門冬)은 A431 cell 및 KHOS-NP cell에 항암작용(抗癌作用)을 보였고, T-cell의 증식(增殖)을 촉진시켰으며, 마우스 복강내(腹腔內) macrophage에서 분비(分泌)되는 NO의 양(量)을 감소시켜고, T cell의 apoptosis에 있어서는 대조군(對照群)에 비하여 유의성(有意性)있게 증가(增加)시켰으며, subpopulation에서는 $T_H$ cell을 증가(增加)시켰다. 그리하여 천문동(天門冬)은 항암작용(抗癌作用) 뿐만아니라 면역세포(免疫細胞)의 증식(增殖)에도 관여하는 약물(藥物)로 인정(認定)할 수 있을 것으로 사료(思料)된다.

• Journal of Microbiology and Biotechnology
• /
• 제19권11호
• /
• pp.1355-1363
• /
• 2009

In the present study, the neuroprotective effects of astaxanthin on $H_2O_2$-mediated apoptotic cell death, using cultured mouse neural progenitor cells (mNPCs), were investigated. To cause apoptotic cell death, mNPCs were pretreated with astaxanthin for 8 h and followed by treatment of 0.3 mM $H_2O_2$. Pretreatment of mNPCs with astaxanthin significantly inhibited $H_2O_2$-mediated apoptosis and induced cell growth in a dose-dependent manner. In Western blot analysis, astaxanthin-pretreated cells showed the activation of p-Akt, p-MEK, p-ERK, and Bcl-2, and the reduction of p-P38, p-SAPK/JNK, Bax, p-GSK3b, cytochrome c, caspase-3, and PARP. Because $H_2O_2$ triggers caspases activation, this study examined whether astaxanthin can inhibit caspases activation in $H_2O_2$-treated mNPCs. After $H_2O_2$ treatment, caspases activities were prominently increased, but astaxanthin pretreatment significantly inhibited $H_2O_2$-mediated caspases activation. Astaxanthin pretreatment also significantly recovered the ATP production ability of $H_2O_2$-treated cells. These findings indicate that astaxanthin inhibits $H_2O_2$-mediated apoptotic features in mNPCs. Inhibition assays with SB203580 ($10\;{\mu}M$, a specific inhibitor of p38) and PD98059 ($10\;{\mu}M$, a specific inhibitor of MEK) clearly showed that astaxanthin can inhibit $H_2O_2$-mediated apoptotic death via modulation of p38 and MEK signaling pathways.

• Journal of Microbiology and Biotechnology
• /
• 제24권2호
• /
• pp.225-235
• /
• 2014

Total 121 lactic acid bacteria were isolated from homemade Inner Mongolia extra hard Hurood cheese. Seven of these strains, identified as Lactobacillus plantarum, were studied for probiotic characteristics. All seven strains survived at pH 3.0 for 3 h, or in the presence of oxgall at 0.3% or 0.6% for 4 h, but their viabilities were affected to different extents at pH 2.0 for 3 h. Strains C37 and C51 showed better adherence to Caco-2 cells, and higher hydrophobicity. The seven L. plantarum strains were different in in vitro free radical scavenging activities and cholesterol-reducing ability. In vivo evaluation of the influence of L. plantarum C37 on the intestinal flora in a mouse model showed strain C37 could increase the viable counts of lactobacilli in feces of mice and decrease the viable counts of enterococci. When L. plantarum C37 was used to prepare probiotic Hurood cheese, it was able to maintain high viable counts (>7.8 log CFU/g) during the whole storage period, but the composition of the cheese was not changed. These results indicate that L. plantarum C37 could be considered as a promising probiotic strain.

• Parasites, Hosts and Diseases
• /
• 제42권2호
• /
• pp.51-56
• /
• 2004

Susceptibilities of 5 different mice strains, including C3H/HeN, BALB/c, C57BL6, FvB and ICR, to Echinostoma hortense infection, was evaluated. The worm expulsion rate, worm size and egg production were observed from 1 to 8 weeks after infection with 30 metacercariae. C3H/HeN and ICR mice showed the highest worm maturation rates. The worm recovery rate and the number of eggs per gram (EPG) of feces was also higher in C3H/HeN and ICR mice than in BALB/c, C57BL6, and FvB mice. It is suggested that E. hortense is highly infectious to ICR and C3H/HeN mice, but not to the other strains of mice. Based on the results obtained, we believe that the susceptibility of different mouse strains to E. hortense infection is dependent on the genetic and immunologic back-ground of mice.

• BMB Reports
• /
• 제56권3호
• /
• pp.172-177
• /
• 2023

BEST family is a class of Ca²⁺-activated Cl- channels evolutionary well conserved from bacteria to human. The human BEST paralogs (BEST1-BEST4) share significant amino acid sequence homology in the N-terminal region, which forms the transmembrane helicases and contains the direct calcium-binding site, Ca²⁺-clasp. But the cytosolic C-terminal region is less conserved in the paralogs. Interestingly, this domain-specific sequence conservation is also found in the BEST1 orthologs. However, the functional role of the C-terminal region in the BEST channels is still poorly understood. Thus, we aimed to understand the functional role of the C-terminal region in the human and mouse BEST1 channels by using electrophysiological recordings. We found that the calcium-dependent activation of BEST1 channels can be modulated by the C-terminal region. The C-terminal deletion hBEST1 reduced the Ca²⁺-dependent current activation and the hBEST1-mBEST1 chimera showed a significantly reduced calcium sensitivity to hBEST1 in the HEK293 cells. And the C-terminal domain could regulate cellular expression and plasma membrane targeting of BEST1 channels. Our results can provide a basis for understanding the C-terminal roles in the structure-function of BEST family proteins.

• Archives of Pharmacal Research
• /
• 제20권3호
• /
• pp.259-263
• /
• 1997

In the preparation of acyclic thymidine nucleoside analogues, $K_2CO_3$(or NaH) treated thymine in DMSO was alkylated with .omega.-chloroalkyl nitrite (Cl-(CH_2)n-CN; n=1, 2, 3, 4) to provide an isomeric mixture of 1-(${\omega}$-cyanoalkyl)thymine (2a-d) and 1,3-bis(${\omega}$-cyanoalkyl)thymine in approximately 5:1 ratios. Reduction of the cyano function 2a-d with $NaBH_{4}/CoCl_{2}$ center dot$ 6H_{2}O$gave the corresponding 1-(${\omega}$aminoalkyl)thymine (3a-d). The newly formed primary amino function in 3a-d was directly reacted with 2-chloroethylisocyanate to afford the 1-[.omega.($N^{I}$-2-chloroethylureido) alkyl]thymine (4a-d) in good yields. Nitrosation of 1-[5-($ N^{I}-2$-chloroethylureido)pentyl] thymine (4d) with glacial acetic acid and dry $NaNO_{2}$-powder in anhydrous $CH_{2}Cl_{2}$gave two types of regioisomeric nitrosoureas, 1-[5-($N^{I}$--chloroethyl-$N^{I}$--nitrosoureido)pentylithymine (5d) and 1-[5-($N^{I}-2$-chloroethyl-N-nitrosoureido)pentyllthymine in approximately 5 :1 ratios. The in vitro cytotoxicity of the synthesized compounds (2a-d, 3a-d, 4a-d and 5a-d) against three cell lines (K-562, P-388 and FM-3A) are measured as $IC^{50}$ values. Compounds 3b and 4c showed moderate activities against all three cell lines, and all other compounds were found to be not active.

• Archives of Pharmacal Research
• /
• 제19권3호
• /
• pp.235-239
• /
• 1996

Six, heretofore undescribed, $5^I-Methyl-5^I-(5-Substituted uracil-1-ylmethyl)-2^I-oxo-3^I-methylenetetrahydrofurans(F, Cl, Br, l, CH_3, H)(6a-f)$were synthesized and evaluated against three cell lines (FM-3A, P-388 and U-937). For the preparation of .alpha.-methylene-.gamma.-butyrolactone bearing 5-substituted uracils (6a-f), the effcient Reformatsky type reaction was employed which involves the treatment of ethyl .alpha.(bromomethyl) acrylate and zinc with the respective 5-substituted uracil-1-ylacetones (5a-f). The acetone derivatives (5a-f) were directly obtained by the respective alkylation reaction of 5-substituted uracils with chloroacetone in the presence of $K_{2}$$CO_{3}$(or NaH). These lactone compounds 6a-f exhibited moderate to significant activity in all of the three cell lines, and 6b, 6c and 6e showed significant antitumor activities (inhibitory concentrations ($IC_{50}$) ranged from 1.3-3.8 .mu.g/ml.

• Clinical and Experimental Reproductive Medicine
• /
• 제27권1호
• /
• pp.1-7
• /
• 2000

Objective: Mammalian embryos undergo changes of energy environment for transfer from oviduct to uterus. Also, the human reproductive organ (oviduct, uterus) contains energy sources of different concentration (oviduct - glucose: 0.5 mM, pyruvate: 0.32 mM, lactate: 10.5 mM; uterus - goucose: 3.15 mM, pyruvate: 0.1mM, lactate: 5.87 mM, respectively). This study was conducted to examine the effect of these energy sources added in DMEM with glutamine on the mouse embryo development. Methods: There was used ICR female mouse. Two cell embryos of mouse are collected by method of 'flushing'. Flushing fluid was used Ham's F-10 added to 20% FBS. The collected 2 cell embryos were cultured in media such as Control (only DMEM), group A and B (DMEM supplemented with 0.5 mM and 3.15 mM glucose), and group C and D (DMEM supplemented with 0.1 mM and 0.32 mM pyruvate), and group E and F (DMEM supplemented with 5.87 mM and 10.5 mM lactate). All experimental media supplemented with 20% hFF, respectively. Pattern of embryo development was observed to interval at 24hr during 96hr. Results : The media with glutamine added glucose (group A: 51.0%; group B: 48.4%) was significantly (p<0.05) higher than other experimental group in development into the morula stage after 24 hr in culture, but not significantly different compared with control and the rate of development into the blastocyst was significantly (p<0.05) low in the both of pyruvate (group C: 7.9% group D: 6.8%) and lactate (group E: 7.1%, group F: 7.1%) treatment group after 48 hr in culture. Development into the blastocyst and hatched balstocyst after 72 hr in culture revealed similarly in control (81.9%) and glucose treatment group (group A: 83.3%, group B: 82.8%). However, development into the hatched and attached blastocyst after 96hr in culture revealed significantly (p<0.05) development in the glucose treatment group (group A: 82.3%, group B: 78.5%) than control (63.2%), and its of pyruvate (group C: 34.1%, group D: 34.1%) and lactate (group E: 25.9%, group F: 33.3%) treatment group were significantly (p<0.05) lower than control similar to previous observations. Conclusion : The glucose added to the DMEM with only glutamine, as energy source, was highly to the rate of development compared with control, but the other energy sources were not, synthetically. Above refer to, the human reproductive organ (oviduct, uterus) contains energy sources of different concentration. Thus, further studies are will examine continuously to effects by interaction of different energy sources in the mouse embryo development, and these results will provide to foundation on the human embryo culture.