• The Korean Journal of Physiology
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• 제25권2호
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• pp.125-131
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• 1991

Inactivation properties of Ca current in the unfertilized eggs of mouse were studied by using the whole cell voltage clamp technique and single microelectrode voltage clamp technique. Membrane potential was held at -80 mV and step depolarization was applied from -50 mV to 50 mV for $200{\sim}500\;ms$. Peak of inward Ca currents was $-2{\sim}-4\;nA$ at a membrane Potentials from -20 mV to 0 mV and outward currents were not observed within the membrane voltage range studied $(-50{\sim}50\;mV)$. Inward currents were fully inactivated within 200 ms after the onset of step depolarization. As the membrane became depolarized, time constant of inactivation (${\tau}$) was decreased but remained around $20{\sim}30\;ms$ beyond 10 mV. When $Ca^{2+}$ was used as a charge earlier, inactivation of inward $Ca^{2+}$ current also occured and time course of inactivation was similar to that of $Ca^{2+}$ currents as charge carrier. In the bathing solution containing high potassium $(131\;mM\;K^+)$, process of inactivation was not changed except a parallel decrease of value for the entire range of membrane potential. Steady-state inactivation of the $current(h_{\infty})$ obtained from the double pulse experiment showed the voltage-dependent change. These results suggested that inactivation of Ca currents in the unfertilized eggs of mouse was voltage-dependent.

• 생명과학회지
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• 제12권1호
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• pp.67-76
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• 2002

Agrobacterium(LBA4404/pBI1121)을 이용하여 형질전환된 애기장대 (Arabidopsis thaliana)를 대상으로 T2, T3, F3세대에서의 도입된 외래 유전자의 비활성화 현상을 조사하였다. Kanamaycin저항성 개체들의 GUS유전자 발현을 분석한 결과, T2세대에서 조사된 12계통 중 5계통에서 GUS 비활성 개체가 관찰되었다 (GUS유전자 비활성율 2.3%). Multi copy T-DNA 계통을 조사한 결과, GUS 비활성 정도가 더욱 심해짐이 관찰되었다 (5.8%). T3 세대에서 single copy T-DNA 계통들은 1.3%의 GUS 비활성율을 보인 반면, multi-copy T-DNA 계통에서의 비활성율은 12.6%로 급격히 증가하였다. 유사한 현상이 형질전환 식물체와 정상개체를 교배하여 생산된 F2 계통에서도 관찰되었다 (비활성율 9.9%). 본 실험으로 식물체에 도입된 외래 유전자가 후대에서의 전이과정동안 점진적으로 비활성화되고, 이 현상은 multi copy T-DNA 계통에서 훨씬 심각함이 밝혀졌다.

• KSBB Journal
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• 제14권2호
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• pp.131-135
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• 1999

원숭이 콩팥세포를 T-flask에서 배양할 때 로타바이러스 감염에 미치는 protease, $CaCl_2$. EGTA, po]ybrene 감염배지내에서 pH의 영향과 formaldehyde에 의한 로타바이러스의 inactivation 정도를 조사하였다. 로타바이러스의 증식은 trypsin이나 clostripain과 같은 protease의 첨가에 의해 크게 향상되었다. $CaCl_2$ 농도가 300 mg/ml이거나 pH가 8인 감염배지에서 로타바이러스의 증식은 각각 8 및 10 배 증가하였다 그러나 EGTA 와 polybrene을 감염배지에 첨가하였을 때 바이러스 증식은 감소하였다. Fonnaldehyde는 로타바이러스 inactivation에 유효하였으며 로타바이러스의 농도는 fomrmaldehyde를 처리하였을 때 1 시간 후 약 53-95% 수준으로 감소하였으며 12 시간 후에는 로타바이러스가 98% 이상 inactivation 되었다.

• 한국축산식품학회지
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• 제33권3호
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• pp.317-324
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• 2013

Radio-frequency driven atmospheric pressure plasma using argon gas was studied in the inactivation of Campylobacter jejuni in order to investigate its applicability. First, the inactivation study was conducted on an agar surface. C. jejuni NCTC11168 was reduced by more than 7 Log CFU after an 88 s treatment. Another strain, ATCC49943, was studied; however, the inactivation was less efficient, with a 5 Log CFU reduction after a 2 min treatment. Then, chicken breast ham was studied at the $10^6$ CFU inoculation level. The inactivation efficiency was much lower for both strains compared to that on the agar plates. C. jejuni NCTC11168 and ATCC49943 were reduced by 3 Log CFU after a 6 min treatment and by 1.5 Log CFU after a 10 min treatment, respectively. The scanning electron microscopy analysis indicated that C. jejuni cells were deformed or transformed into coccoid form under the plasma treatment. During the plasma treatment, the temperature of the samples did not rise above $43^{\circ}C$, suggesting that heat did not contribute to the inactivation. Meanwhile, water activity significantly decreased after a 10 min treatment (p<0.05). This study conveyed that radio-frequency atmospheric pressure plasma can effectively inactivate C. jejuni with strain-specific variation.

• The Korean Journal of Physiology and Pharmacology
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• 제11권5호
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• pp.183-188
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• 2007

Using BHK cells with stable expression of cardiac $Na^+/Ca^{2+}$ exchanger(BHK-NCX1), reverse mode(i.e. $Ca^{2+}$ influx mode) of NCX1 current was recorded by whole-cell patch clamp. Repeated stimulation of reverse NCX1 produced a cytosolic $Ca^{2+}$-dependent long-term inactivation of the exchanger activity. The degrees of inactivation correlated with NCX1 densities of the cells and were attenuated by reduced $Ca^{2+}$ influx via the reverse exchanger. The inactivation of NCX1 was attenuated by(i) inhibition of $Ca^{2+}$ influx with reduced extracellular $Ca^{2+}$, (ii) treatment with NCX1 blocker($Na^{2+}$), and (iii) increase of cytoplasmic $Ca^{2+}$ buffer(EGTA). In BHK-NCX1 cells transiently expressing TRPV1 channels, $Ca^{2+}$ influx elicited by capsaicin produced a marked inactivation of NCX1. We suggest that cytoplasmic $Ca^{2+}$ has a dual effect on NCX1 activities, and that allosteric $Ca^{2+}$ activation of NCX1 can be opposed by the $Ca^{2+}$-dependent long-term inactivation in intact cells.

• 한국식품영양학회지
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• 제34권1호
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• pp.26-35
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• 2021

Myrosinases (thioglucosidases) catalyze the hydrolysis of a class of compounds called glucosinolates, of which the aglycones show various biological functions. It is often necessary to minimize the loss of myrosinase activity during thermal processing of cruciferous vegetables. Myrosinase was isolated from a popular spice, white mustard (Sinapis alba), and its thermal inactivation kinetics was investigated. The enzyme was extracted from white mustard seeds and purified by a sequential processes of ammonium sulfate fractionation, Concanavalin A-Sepharose column chromatography, and gel permeation chromatography. At least three isozymes were revealed by Concanavalin A-Sepharose column chromatography. The purity of the major myrosinase was examined by native polyacrylamide gel electrophoresis and on-gel activity staining with methyl red. The molecular weight of the major enzyme was estimated to be 171 kDa. When the consecutive step model was used for the thermal inactivation of the major myrosinase, its inactivation energy was 44.388 kJ/mol for the early stage of destruction and 32.019 kJ/mol for the late stage of destruction. When the distinct two enzymes model was used, the inactivation energy was 77.772 kJ/mol for the labile enzyme and 95.145 kJ/mol for the stable enzyme. The thermal inactivation energies lie within energy range causing nutrient destruction on heating.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2001년도 학술 발표회 진행표 및 논문초록
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• pp.26-26
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• 2001

Inactivation of N-type calcium current has been reported to be both voltage dependent and Ca$\^$2＋/ dependent. We have investigated the effects of Ba$\^$2＋/ and Ca$\^$2＋/ on N-channel inactivation in rat superior cervical ganglion neurons using the whole cell configuration of patch clamp technique. Inactivation was larger in Ca$\^$2＋/ than in Ba$\^$2＋/ even with 20 mM BAPTA.(omitted)

• 한국의학물리학회지:의학물리
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• 제15권4호
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• pp.220-227
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• 2004

N형 칼슘통로의 비활성화기전에 관하여는 아직까지도 막전압의존성 기전과 칼슘의존성 기전간에 논란이 계속되고 있다. 2003년에 의학물리에 발표한 논문1)에서 본 연구자는 N형 칼슘통로의 비활성화 기전은 2가지 성분 -빠른 성분과 느린 성분을 가지고 있고 빠른 성분은 칼슘의존적이 아니며 오직 느린 성분만이 칼슘의존적일 가능성을 제시하였다. 본 논문에서는 막전압의존성 기전이 옳건 칼슘의존성 기전이 옳건 간에 세포 신호전달 체계로서 비활성화와 연계된 기전이 필요하므로 이러한 맥락에서 인산화 기전을 연구하였다. 흰쥐 경동맥 결절뉴론을 단일 세포로 얻은 후 whole cell patch clamp technique를 사용하여 N형 칼슘전류를 기록하고 대조 세포내액을 사용하였을 때와 phosphatase inhibitor인 okadaic acid를 포함한 세포내액을 사용하였을 때의 차이를 비교하였다. Okadaic acid에 의하여 비활성화정도가 증가되었고 이러한 okadaic acid 효과는 주로 N형 통로를 통하여 영향을 미침을 N형 칼슘통로 억제제인 $\omega$-conotoxin GVIA를 사용함으로써 확인하였다. Okadaic acid에 의한 비활성화 증가 효과는 protein kinase를 비특이적으로 억제하는 staurosporine에 의하여 억제되었고 또한 calmodulin dependent protein kinase의 특이적 억제제인 lavendustin C에 의하여 억제되었으므로 인산화과정이 N형 칼슘통로 비활성화와 관련되어 있고 특히 calmodulin을 통한 인산화과정이 주로 관여함을 확인하였다. 본 연구자가 발표한 선행논문1)에 의해 외부의 2가 양이온에 의해 빠른 비활성화가 진행되며, 본 논문에 의하여 인산화과정에 의해 빠른 비활성화가 촉진된다는 사실이 확인되었다. 그러나 본 연구결과만으로는 인산화과정이 비활성화 자체라고는 볼 수 없으며 단지 인산화과정에 의해 비활성화가 가속되었다고 해석할 수 밖에 없다. 인산화과정이 비활성화자 체인지 여부는 2가 양이온이 칼슘통로에 작용하는 결합부위에 관한 연구 및 인산화 부위가 칼슘통로인지 아니면 다른 조절 부위인지 여부를 확인할 수 있는 연구가 진행되어야 확실히 알 수 있을 것이다.

• 한국식품과학회지
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• 제47권1호
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• pp.70-74
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• 2015

생식제조에 사용되는 곡류의 미생물 오염을 저감하기 위하여 코로나방전플라즈마제트(CDPJ)를 사용하여 곡류 표면의 미생물을 살균을 시도하고, CDPJ 처리에 따른 곡류 품질변화 여부를 조사하였다. 곡류의 초기 오염도는 $1.7{\times}10^3-9.9{\times}10^5CFU/g$ 정도를 보였다. 곡류에 대한 CDPJ의 살균력은 1-1.5 A 범위에서 전류세기에 따라 증가하였으며, 토출거리 25 mm에서 가장 양호한 살균 효과를 나타냈다. 곡류 미생물의 CDPJ 살균패턴은 의사 1차반응 모델인 Singh-Heldman 모델에 부합하였고, 세균이 진균에 비해 더 민감한 살균효과를 보였다. 곡류별로는 백미가 가장 잘 살균되었고, 압맥, 현미 순으로 살균효과가 낮아졌다. 처리시간 10분 이하의 처리는 곡류의 TBA가에 영향을 주지 않아 CDPJ 처리는 생식제조용 곡류의 지방특성을 변화시키지 않고 미생물을 저감함으로써 생식의 위생 향상에 활용될 수 있을 것으로 기대된다.

• BMB Reports
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• 제30권2호
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• pp.113-118
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• 1997

Aspartase from Hafnia alvei was inactivated by N-ethylmaleimide (NEM) and 5,5' -Dithiobis-(2-znitrobenzoic acid) (DTNB) following pseudo-first order kinetics. Their apparent reaction orders were 0.83 and 0.50 for NEM and DTNB modifications, respectively, indicating that inactivation was due to a sulfhydryl group in the active site of aspartase and participation of the sulfhydryl group in an essential step in the catalytic reaction. When aspartase was modified by DTNB, the enzyme activity was restored by dithiothreitol treatment, indicating that cysteine residuetsl islarel possibly at or near the active site. The pH-dependence of the inactivation rate by NEM suggested that an amino acid residue having pK value of 8.3 was involved in the inactivation. When aspartase was incubated with NEM and L-aspartate together, L-aspartate markedly protected the enzyme from inactivation by NEM, but the other reagents used did not.