• The Korean Journal of Physiology and Pharmacology
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• 제14권2호
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• pp.83-89
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• 2010

In this study, we studied whether hydrogen sulfide ($H_2S$) has an effect on the pacemaker activity of interstitial cells of Cajal (ICC), in the small intestine of mice. The actions of $H_2S$ on pacemaker activity were investigated using whole-cell patch-clamp technique, intracellular $Ca^{2+}$ analysis at $30^{\circ}C$ and RT-PCR in cultured mouse intestinal ICC. Exogenously applied sodium hydrogen sulfide (NaHS), a donor of hydrogen sulfide, caused a slight tonic inward current on pacemaker activity in ICC at low concentrations (50 and $100{\mu}m$), but at high concentration ($500{\mu}m$ and 1 mM) it seemed to cause light tonic inward currents and then inhibited pacemaker amplitude and pacemaker frequency, and also an increase in the resting currents in the outward direction. Glibenclamide or other potassium channel blockers (TEA, $BaCl_2$, apamin or 4-aminopydirine) did not have an effect on NaHS-induced action in ICC. The exogenous application of carbonilcyanide p-triflouromethoxyphenylhydrazone (FCCP) and thapsigargin also inhibited the pacemaker activity of ICC as NaHS. Also, we found NaHS inhibited the spontaneous intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) oscillations in cultured ICC. In doing an RT-PCR experiment, we found that ICC enriched population lacked mRNA for both CSE and CBS, but was prominently detected in unsorted muscle. In conclusion, $H_2S$ inhibited the pacemaker activity of ICC by modulating intracellular $Ca^{2+}$. These results can serve as evidence of the physiological action of $H_2S$ as acting on the ICC in gastrointestinal (GI) motility.

• Reproductive and Developmental Biology
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• 제38권4호
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• pp.147-158
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• 2014

Differentiated nuclei can experimentally be returned to an undifferentiated embryonic status after nuclear transfer (NT) to unfertilized metaphase II (MII) oocytes. Nuclear reprogramming is triggered immediately after somatic cell nucleus transfer (SCNT) into recipient cytoplasm and this period is regarded as a key stage for optimizing reprogramming. In a recent study (Dai et al., 2010), use of m-carboxycinnamic acid bishydroxamide (CBHA) as a histone deacetylase inhibitor during the in vitro early culture of murine cloned embryos modifies the acetylation status of somatic nuclei and increases the developmental competence of SCNT embryos. Thus, we examined the effects of CBHA treatment on the in vitro preimplantation development of porcine SCNT embryos and on the acetylated status of histone H3K9 on cloned embryos at the zygote stage. We performed the three groups SCNT: SCNT (NT), CBHA treatment at the porcine fetus fibroblast cells (PFFs) used as donor cells prior to SCNT (CBHA-C) and CBHA treatment at the porcine SCNT embryos during the in vitro early culture after oocyte activation (CBHA-Z). The PFFs were treated with a $15{\mu}M$ of CBHA (8 h) for the early culture and the porcine cloned embryos were treated with a $100{\mu}M$ concentration of CBHA during the in vitro early culture (10 h). Cleavage rates and development to the blastocyst stage were assessed. No significant difference was observed the cleavage rate among the groups (82.6%, 76.4% and 82.2%, respectively). However, the development competence to the blastocyst stage was significantly increased in CBHA-Z embryos (22.7%) as compared to SCNT and CBHA-C embryos (8.6% and 4.1%)(p<0.05). Total cell numbers and viable cell numbers at the blastocyst stage of porcine SCNT embryos were increased in CBHA-Z embryos as compared to those in CBHA-C embryos (p<0.05). Signal level of histone acetylation (H3K9ac) at the zygote stage of SCNT was increased in CBHA-Z embryos as compared to SCNT and CBHA-C embryos. The results of the present study suggested that treatment with CBHA during the in vitro early culture (10 h) had significantly increased the developmental competence and histone acetylation level at the zygote stage.

• 한국재료학회지
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• 제10권3호
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• pp.246-252
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• 2000

기계적 합금화 공정을 이용하여 제조한 $Bi_2(Te_{0.85}Se_{0.15})_3$ 가압소결체의 가압소결온도에 따른 열전특성을 분석하였다. $Bi_2(Te_{0.85}Se_{0.15})_3$ 가압소결체는 $300^{\circ}C$에서 $550^{\circ}C$ 범위의 가압소결온도에 무관하게 n형 전도를 나타내었다. $Bi_2(Te_{0.85}Se_{0.15})$ 합금분말을 (50% $H_2+50%$ Ar) 분위기에서 환원처리시, 분말 표면의 산화층 제거 및 과잉 Te 공격자의 소멸에 기인한 전자 농도의 감소로 가압소결체의 Seebeck 계수가 양의 값으로 변화하였다. $450^{\circ}C$ 이상의 온도에서 가압소결시 가압소결온도의 증가에 따라 $Bi_2(Te_{0.85}Se_{0.15})$ 합금의 성능지수가 증가하였으며, $550^{\circ}C$에서 가압소결시 $1.92{\times}10^{-3}/K$의 최대성능지수를 얻을 수 있었다.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.127-134
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• 2010

This study was to investigate the effect of flavonoid treatment on in vitro development of bovine somatic cell nuclear transfer (SCNT) embryos, and their pregnancy and delivery rate after embryo transfer into recipient. In experiment 1, to optimize the flavonoid concentration, parthenogenetic day 2 ($\geq$ 2-cell) embryos were cultured in 0 (control), 1, 10 and $20\;{\mu}M$ flavonoid for 6 days. In the results, in vitro development rate was the highest in $10\;{\mu}M$ flavonoid group (57.1%) among treatment groups (control, 49.5%; $1\;{\mu}M$, 54.2%; $20\;{\mu}M$, 37.5%), and numbers of total and ICM cells were significantly (p<0.05) higher in $10\;{\mu}M$ flavonoid group than other groups. We found that $10\;{\mu}M$ flavonoid treatment can significantly (p<0.05) decrease the apoptotic index and derive high expression of anti-oxidant, anti-apoptotic, cell growth and development marker genes such as Mn-SOD, Survivin, Bax inhibitor, Glut-5, In-tau, compared to control group. In experiment 2, to produce the cloned Jeju Black Cattle, beef quality index grade 1 bull somatic cells were transferred into enucleated bovine MII oocytes and reconstructed embryos were cultured in $10\;{\mu}M$ flavonoid added medium. When the in vitro produced day 7 or 8 SCNT blastocysts were transferred into a number of recipients, $10\;{\mu}M$ flavonoid treatment group presented higher pregnancy rate (10.2%, 6/59) than control group (5.9%, 2/34). Total three cloned Jeju Black calves were born. Also, two cloned calves in $10\;{\mu}M$ flavonoid group were born and both were all healthy at present, while the one cloned calf born in control group was dead one month after birth. In addition, when the result of short tandem repeat marker analysis of each cloned calf was investigated, microsatellite loci of 11 numbers matched genotype between donor cell and cloned calf tissue. These results demonstrated that the flavonoid addition in culture medium may have beneficial effects on in vitro and in vivo developmental capacity of SCNT embryos and pregnancy rate.

• Journal of Periodontal and Implant Science
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• 제41권1호
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• pp.17-22
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• 2011

Purpose: Nitric oxide (NO) has been known as an important regulator of osteoblasts and periodontal ligament cell activity. This study was performed to investigate the relationship between NO-mediated cell death of human periodontal ligament fibroblasts (PDLFs) and N-methyl-D-aspartic acid (NMDA) receptor antagonist (+)-5-methyl-10, 11-dihydro-5H-dibenzo[a,d]cyclohepten-5, 10-imine hydrogen maleate (MK801). Methods: Human PDLFs were treated with various concentrations (0 to 4 mM) of sodium nitroprusside (SNP) with or without $200\;{\mu}M$ MK801 in culture media for 16 hours and the cell medium was then removed and replaced by fresh medium containing MTS reagent for cell proliferation assay. Western blot analysis was performed to investigate the effects of SNP on the expression of Bax, cytochrome c, and caspase-3 proteins. The differences for each value among the sample groups were compared using analysis of variance with 95% confidence intervals. Results: In the case of SNP treatment, as a NO donor, cell viability was significantly decreased in a concentration-dependent manner. In addition, a synergistic effect was shown when both SNP and NMDA receptor antagonist was added to the medium. SNP treated PDLFs exhibited a round shape in culture conditions and were dramatically reduced in cell number. SNP treatment also increased levels of apoptotic marker protein, such as Bax and cytochrome c, and reduced caspase-3 in PDLFs. Mitogen-activated protein kinase signaling was activated by treatment of SNP and NMDA receptor antagonist. Conclusions: These results suggest that excessive production of NO may induce apoptosis and that NMDA receptor may modulate NO-induced apoptosis in PDLFs.

• 대한환경공학회지
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• 제36권5호
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• pp.352-358
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• 2014

수중 내 함유된 고농도 perchlorate의 환원가능성을 perchlorate 환원 미생물인 신균주 Rhodococcus sp. YSPW01과 YSPW02로 검토하였다. Perchlorate 환원 미생물은 혐기소화조 슬러지에서 분리 배양하였으며, perchlorate 환원능을 검토하기 위해 전자공여체인 acetate를 사용하였다. YSPW01과 YSPW02는 perchlorate $82mg\;L^{-1}$와 acetate $550mg\;L^{-1}$에서 회분식 실험을 수행하였을 때, 반응 26시간과 9시간 후 각각 정상상태에 도달하였다. 이 때 perchlorate 환원은 8, 7시간 이내에 초기농도 $82mg\;L^{-1}$에서 검출한계 이하($10{\mu}g\;L^{-1}$)까지 제거되었다. 반응조 내에 acetate:perchlorate (w:w)비를 1:1에서 5:1로 증가한 결과, perchlorate의 제거 속도는 YSPW01과 YSPW02 모두 2.1, $3.2mg\;L^{-1}h^{-1}$에서 15, $15.5mg\;L^{-1}h^{-1}$로 약 4.5배 증가하였고, 최종 perchlorate는 검출한계 이하까지 제거되었다. 본 연구결과, Rhodococcus sp. YSPW01 및 YSPW02는 고농도 perchlorate 제거에 적합하며, 신균주를 고농도 perchlorate 제거를 위한 생물학적 처리공정에 응용가능 할 것으로 판단된다.

• 대한환경공학회지
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• 제22권3호
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• pp.405-415
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• 2000

전자공여체로 황 입자를 충전시킨 상향류 고정상에서 유입수의 $NO_3{^-}$-N 농도를 일정하게 유지하고 체류시간과 온도를 변화시키면서 독립영양 탈질실험을 실시한 결과, 체류시간 및 온도가 감소함에 따라 처리수의 $NO_2{^-}$-N 및 발생가스의 $NO_2$ 농도가 증가하면서 탈질효율이 악화되고 가스발생량은 감소하였다. 제거된 $NO_3{^-}$-N 1g당 소모된 알칼리도의 양은 실험조건에 따라 차이를 나타내어, 체류시간 영향 실험에서는 3.44~5.71 g. 평균값 4.67 g로 이론값인 4.57 g에 근사하였으나, 온도 영향 실험에서는 6.58~13.41 g, 평균값 9.12g은 이론값의 2배 정도되었다. 제거된 $NO_3{^-}$-N 1 g당 생성된 $SO_4{^{2-}}$의 양은 체류시간 영향 실험에서는 6.99~9.69 g, 평균 8.20 g이었으며, 온도 영향 실험에서는 6.44~8.16 g, 평균 7.51g으로 이론값인 7.54 g에 가깝게 나타났다. 반응조 길이 방향에 따른 탈질은 반응속도상수가 0.1648/hr인 1차반응으로 그리고 황산염 생성은 반응속도상수가 241/hr인 0차반응으로 나타났으며, 온도보정계수는 평균 1.130으로 계산되었다. 그러나 질소수지를 따진 결과 이론치와 실측치간에 큰 차이를 보여 이의 규명을 위한 연구가 요망된다.

• The Korean Journal of Physiology and Pharmacology
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• 제4권5호
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• pp.369-378
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• 2000

This study was undertaken to investigate an involvement of nitroxergic innervation in gastric smooth muscle of rat. Isometric tension study, the measurement of single cell length, NADPH diaphorase stain of smooth muscle layers and neuronal nitric oxide synthase (nNOS) western blotting were performed. Sodium nitroprusside (SNP), a nitric oxide donor, relaxed the muscle strips precontracted by acetylcholine (ACh) in a concentration-dependent manner. Pretreatment of L-arginine decreased the contraction induced by electric field stimulation (EFS). Pretreatment of $N^G-nitro-L-arginine$ methyl ester (L-NAME), a NOS inhibitor, increased the EFS-induced contractions. LY 83583, a guanylate cyclase (GC) inhibitor, reversed the inhibitory actions of L-arginine on the muscle contractions. The effects of L-Arginine, L-NAME and LY 83583 on ACh-induced contractions were not significant. L-arginine reduced the EFS-induced contraction in circular muscle, whereas L-NAME enhanced the EFS-induced contraction in longitudinal strips. By EFS, the phasic contractions appeared approximately $20{\sim}25$ seconds later. L-NAME significantly shortened the delay time to about $2{\sim}3$ seconds. In single cell study, ACh contracted gastric smooth muscle cells, SNP relaxed the cells, and the latter also inhibited the ACh-induced contraction. LY 83583 enhanced the ACh-induced contraction and antagonized SNP-induced relaxation. NADPH diaphorase activity was assessed by a histochemistry, nitroblue tetrazolium (NTB) staining. Positive staining was observed in both circular and longitudinal muscle layers. L-arginine increased the staining, while L-NAME decreased the staining. Western blotting for nNOS proved the presence of nNOS in rat gastric smooth muscle. EFS and additional $Ca^{2+}$ increased nNOS protein expression. These results suggest that in rat stomach, both circular and longitudinal muscle layers are innervated with nitroxergic nerves which relax the gastric smooth muscle via NO-cGMP pathway.

• The Korean Journal of Physiology and Pharmacology
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• 제19권6호
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• pp.507-514
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• 2015

Nitric oxide (NO) is important in the regulation of bone remodeling, whereas high concentration of NO promotes cell death of osteoblast. However, it is not clear yet whether NO-induced autophagy is implicated in cell death or survival of osteoblast. The present study is aimed to examine the role of NO-induced autophagy in the MC3T3-E1 cells and their underlying molecular mechanism. The effect of sodium nitroprusside (SNP), an NO donor, on the cytotoxicity of the MC3T3-E1 cells was determined by MTT assay and expression of apoptosis or autophagy associated molecules was evaluated by western blot analysis. The morphological observation of autophagy and apoptosis by acridine orange stain and TUNEL assay were performed, respectively. Treatment of SNP decreased the cell viability of the MC3T3-E1 cells in dose- and time-dependent manner. SNP increased expression levels of p62, ATG7, Beclin-1 and LC3-II, as typical autophagic markers and augmented acidic autophagolysosomal vacuoles, detected by acridine orange staining. However, pretreatment with 3-methyladenine (3MA), the specific inhibitor for autophagy, decreased cell viability, whereas increased the cleavage of PARP and caspase-3 in the SNP-treated MC3T3-E1 cells. AMP-activated protein kinase (AMPK), a major autophagy regulatory kinase, was activated in SNP-treated MC3T3-E1 cells. In addition, pretreatment with compound C, an inhibitor of AMPK, decreased cell viability, whereas increased the number of apoptotic cells, cleaved PARP and caspase-3 levels compared to those of SNP-treated MC3T3-E1 cells. Taken together, it is speculated that NO-induced autophagy functions as a survival mechanism via AMPK activation against apoptosis in the MC3T3-E1 cells.

• 대한전기학회:학술대회논문집
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• 대한전기학회 1988년도 추계학술대회 논문집 학회본부
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• pp.372-375
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• 1988

The III-V ternary alloy semiconductor $In_{l-x}Ga_{x}As$ were grown by the temperature Gradient of $0.60{\leq}x{\leq}0.98$. The electrical properties were investigated by the Hall effect measurement with the Van der Pauw method in the temperature range of $90{\sim}300K$. $In_{l-x}Ga_{x}As$ were revealed n-type and the carrier concentration at 300K were in the range of $9.69{\times}10^{16}cm^{-3}{\sim}7.49{\times}10^{17}cm^{-3}$. The resistivity was increased and the carrier mobility was decreased with increasing the composition ratio. The optical energy gap determined by optical transmission were $20{\sim}30meV$ lower than theoretical valves on the basis of absorption in the conduction band tail and it was decreased with increasing the temperature by the Varshni rule. In the photoluminescence of undoped $In_{l-x}Ga_{x}As$ at 20K, the main emission was revealed by the radiative recombination of shallow donor(Si) to acceptor(Zn) and the peak energy was increased with increasing the composition, X. The diffusion depth of Zn increases proportionally with the square root of diffusion time, and the activation energy for the Zn diffusion into $In_{0.10}Ga_{0.90}As$ was 2.174eV and temperatures dependence of diffusion coefficient was D = 87.29 exp(-2.174/$K_{B}T$). The Zn diffusion p-n $In_{x}Ga_{x}As$ diode revealed the good rectfying characteristics and the diode factor $\beta{\approx}2$. The electroluminescence spectrum for the Zn-diffusion p-n $In_{0.10}Ga_{0.90}As$ diode was due to radiative recombation between the selectron trap level(${\sim}140meV$) and Zn acceptor level(${\sim}30meV$). The peak energy and FWHM of electroluminescence spectrum at 77K were 1.262eV and 81.0meV, respectively.