• Title/Summary/Keyword: inactivation.

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Voltage-Dependent Inactivation of Calcium Currents in the Mouse Eggs

  • Park, Young-Geun;Yang, Young-Seon;Yum, Myung-Kul;Hong, Seong-Geun
    • The Korean Journal of Physiology
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    • v.25 no.2
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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.

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The increased GUS gene inactivation over generation in Arabidopsis transgenic lines (애기장대 형질전환 식물체의 세대경과에 따른 GUS유전자의 비활성화에 관한 연구)

  • Park, Soon-Ki
    • Journal of Life Science
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    • v.12 no.1
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    • pp.67-76
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    • 2002
  • The effect of transgene inactivation in T2, T3 and F2 generations was analyzed in progeny seedlings which had been generated by Agrobacterium (LBA4404/pBI121)-mediated transformation in Arabidopsis thaliana. In a system which investigated in the expression of $\beta$-glucuronidase(GUS)gene in kanamycin-resistant (ke $n^{R}$)seedlings, GUS inactivated seedlings were observed in 5 of 12 tested lines of T2 generation and the frequency of GUS inactivation was approximately 2.3%. Lines with multi-copies of T-DNA exhibited severe GUS gene inactivation with the frequency of 5.8% in T2 generation. In T3 generation lines exhibited GUS gene inactivation with the frequency of 1.3%. In contrast, inactivation increased dramatically up to 12.6% in multi-copy T-DNA line. A similar phenomenon was also found in F2 progeny from a transgenic line which had been crossed with wild-type Arabidopsis plant, WS-O (GUS gene inactivation frequency 9.9%). These results indicate that the foreign gene introduced into the plant was inactivated progressively in its transmission during subsequent generations and the transgenic line with multi-copies of T-DNA tended to show more increased inactivation.

Studies on the Infection Condition and Formaldehyde Inactivation of Human Rotavirus Wa (사람 로타바이러스 Wa의 감염 조건 및 formaldehyde에 의한 inactivation에 대한 연구)

  • 장경화;박종화;박창호;정인식
    • KSBB Journal
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    • v.14 no.2
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    • pp.131-135
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    • 1999
  • Effects of protease, $CaCl_2$ EGTA, polybrene, medium pH, and formaldehyde on the infection and inactivation of human rotavirus Wa were investigated using T-flask culture of monkey kidney MA-104 cells. Rotavirus titer was improved by the addition of trypsin or clostripain. Rotavirus titer was increaeed 8 and 10 time sin the infection medium supplemented with 300 $\mu\textrm{g}$/mL of $CaCl_2$ and in the medium adjusted its pH to 8, respectively. However, addition of EGTA or polybrene to the medium decreased rotavirus titer. Rotavirus titer was reduced to 53-95% of the initial value at 1 hr after formaldehyde treatment. Furthermore, rotavirus was inactivated more than 98% at 12 hrs after formaldehyde treatment.

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Inactivation of Campylobacter jejuni using Radio-frequency Atmospheric Pressure Plasma on Agar Plates and Chicken Hams

  • Kim, Joo-Sung;Lee, Eun-Jung;Cho, Eun-Ah;Kim, Yun-Ji
    • Food Science of Animal Resources
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    • v.33 no.3
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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.

[ $Ca^{2+}$ ]-dependent Long-term Inactivation of Cardiac $Na^+/Ca^{2+}$ Exchanger

  • Lee, Jee-Eun;Kang, Tong-Mook
    • The Korean Journal of Physiology and Pharmacology
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    • v.11 no.5
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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.

Thermal Inactivation of Myrosinase from White Mustard Seeds

  • Ko, Young Hwan;Lee, Ran
    • The Korean Journal of Food And Nutrition
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    • v.34 no.1
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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.

Inactivation of N-Type Calcium Current in Rat Sympathetic Neurons

  • Lee, Mi-Sun;Goo, Yong-Sook
    • Proceedings of the Korean Biophysical Society Conference
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    • 2001.06a
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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)

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Phosphorylation as a Signal Transduction Pathway Related with N-channel Inactivation in Rat Sympathetic Neurons (N형 칼슘통로 비활성화와 연계된 세포 신호전달 체계로서의 인산화과정)

  • Lim Wonil;Goo Yong Sook
    • Progress in Medical Physics
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    • v.15 no.4
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    • pp.220-227
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    • 2004
  • In N-type $Ca^{2+}$ channels, the mechanism of inactivation - decline of inward current during a depolarizing voltage step- is still controversial between voltage-dependent inactivation and $Ca^{2+}$ -dependent inactivation. In the previous paper we demonstrated that fast component of inactivation of N-type calcium channels does not involve classic $Ca^{2+}$ -dependent mechanism and the slowly inactivating component could result from a $Ca^{2+}$ -dependent process. However, there should be signal transduction pathway which enhances inactivation no matter what the inactivation mechanism is. We have investigated the effect of phosphorylation on calcium channels of rat sympathetic neurons. Intracellular dialysis with the phosphatase inhibitors okadaic acid markedly enhanced the inactivation. The rapidly inactivating component is N-type calcium current, which is blocked by $\omega$-conotoxin GVIA. Staurosporine, a nonselective protein kinase inhibitor, prevented the action of okadaic acid, suggesting that protein phosphorylation is involved. More specifically lavendustin C, inhibitor of CaM kinase II, prevented the action of okadaic acid, suggesting that calmodulin dependent pathway is involved in inactivation process. It is not certain to this point whether phosphorylation process is inactivation itself. Molecular biological research regarding binding site should be followed to address the question of how the divalent cation binding site is related to phoshorylation process.

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Microbial Inactivation of Grains Used in Saengshik by Corona Discharge Plasma Jet (코로나방전플라즈마제트를 이용한 생식용 곡류의 미생물 저감)

  • Youn, Geum-A;Mok, Chulkyoon
    • Korean Journal of Food Science and Technology
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    • v.47 no.1
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    • pp.70-74
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    • 2015
  • Inactivation of microorganisms in grains used for saengshik, a formulated health food, was attempted by corona discharge plasma jet (CDPJ). The initial microbial counts of the grains were in the range of $1.7{\times}10^3-9.9{\times}10^5CFU/g$. The CDPJ-inactivation effect was increased with electric current in the range of 1-1.5 A. Regarding span length between the tips of the electrodes and the treatment surface, the highest inactivation effect was observed at 25 mm. The inactivation pattern fitted well to the Singh-Heldman model. Bacteria were more labile to the CDPJ inactivation than yeasts and molds. Among tested grains, white rice showed the highest sterility followed by pressed barley and brown rice. Despite the inactivation by plasma, the thiobarbituric acid content of the grains remained unchanged over 10 min of treatment. Our results indicated the potential of the CDPJ treatment to improve the hygiene of saengshik products with no remarkable changes in lipid quality.

Chemical Modification of Cysteine Residues in Hafnia alvei Aspartase by NEM and DTNB

  • Shim, Joon-Bum;Kim, Jung-Sung;Yoon, Moon-Young
    • BMB Reports
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    • v.30 no.2
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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.

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