• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.2
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• pp.292-298
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• 1999

Myofibril and actomyosin were prepared from red muscle and white muscle of fresh water fish and sea water fish, and their biochemical characteristics and SDS PAGE patterns of myofibril were compared. SDS PAGE analysis showed that electrophoretic patterns of myofibril were similar be tween white muscle and red muscle, while difference of 30kDa component of myofibril was detected between fresh water fish and sea water fish. When myofibril were treated with trypsin, difference in hydrolysis of heavy chain was observed between white muscle and red muscle. In activities of Ca ATPase, Mg ATPase, EDTA ATPase and ATPase activity pH curve, myofibrillar protein from fresh water fish showed higher specific activity than those from sea water fish.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.153-154
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• 2003

운동여부와 운동횟수 그리고 어체 크기에 대한 양식산 어종의 품질판정을 위한 지표를 검색하기 위하여 효소적 방법 즉 ATP 관련화합물과 양식산 활어의 건강한 상태와 칼슘의 노출, 염도의 감소, 산소의 부족, 궁핍 등과 같은 스트레스를 받은 상태를 판단하는 adenylate energy charge (AEC)를 근육의 품질판정을 위한 지표로써 조사하였다. 운동사육구와 비운동사육구에서의 참돔육에 대한 ATP 관련화합물의 변화는 운동 0일에는 ATP함량은 8.62 μmol/g이었지만 20일차에는 8.59 μmol/g으로 운동기간이 길어짐에 따라 ATP의 함량은 약간의 감소를 보였다. 이는 운동이라는 자극요인으로 인하여 ATP의 소모가 촉진되었을 것으로 판단된다. 또한 운동 0일차의 ADP함량은 0.72 μmo1/g이었지만 20일차에는 0.93μmol/g로 ATP의 소모만큼 약간의 증가를 보이고 있다. 반면, 비운동사육구에서 0일차의 ATP함량은 5.78 μmol/g이었지만 20일차에서는 6.31 μmol/g로 운동사육구와 대조적으로 기간이 길어짐에 따라 ATP함량이 약간의 증가를 보여주고 있다. 생리적으로 동물에 가혹한 환경에 두거나 스트레스를 가하면 adenylate energy charge (AEC)가 영향을 받아 변화게 된다. AEC는 (ATP ＋ ½ ADP)/(ATP ＋ ADP + AMP)의 비율로 정의되는데, AEC의 범위는 0∼1사이로써 0은 ATP 관련화합물이 AMP로 전환된 상태를 말하며, 1은 모든 ATP 관련 화합물이 ATP로 존재하는 것을 의미한다. 그러므로, 이들결합의 상관적인 정도는 개개의 세포에서 에너지의 효용성을 측정하는 것으로 이용되고 있다. AEC 수치는 운동수조에서 0.91∼0.92 건강상태가 좋았지만 비운동수조는 0.87∼0.88로 운동수조에 비해 값이 낮게 나왔다. 운동횟수에 관계없이 운동시킨 어육의 AEC는 0.90∼0.91로 건강한 상태였다. 어체크기에 따른 ATP 관련화합물의 변화는 0.7∼0.9kg의 크기를 가진 참돔에서는 ATP 함량이 7.88 μmole/g, 1.5∼l,7kg의 크기를 가진 참돔에서는 ATP함량이 9.61μmole/g로 많았고 다른 크기에서는 큰 차이를 나타내지 않았다. 그리고 ADP함량은 어체의 크기가 증가할수록 함량은 감소하였다. 그러나, 어체의 크기에 따른 ATP관련화합물의 변화는 거의 없었다. 건강도의 지표인 AEC는 0.89∼0.94로 전체적으로 건강상태로 나타났다. 하루 2회 운동시킨 육에서의 ATP함량은 0일차에 8.75 μmole/g, 10일차에 8.46 μmole/g, 20일차에 8.51μmole/g로 약간의 감소를 보여준다. ADP함량은 일까지는 증가하지만 그 이후에는 큰 변화를 보이지 않는다. 6시간 운동시킨 그룹의 ATP함량은 0일차에 8.63 μmole/g, 5일차에 8.52 μmole/g, 10일차에 8.4μmole/g, 20일차에 8.51 μmole/g로 10일까지는 감소하지만 그 이후에는 큰 변화를 보이지 않는다. ADP함량은 0일차에 0.72 μmole/g, 10일차 0.84 μmole/g, 20일차에 0.83 μmole/g으로 10일차까지는 약간 증가하지만 그 이후에는 큰 변화를 보이지 않았다. 참돔의 근원섬유에 있어서 Ca/sup 2+/이 존재할 때 Mg/sup 2+/-ATPase 활성의 변화는 운동시킨 참돔 근원섬유의 Mg/sup 2+/-ATPase 활성은 0.35μmol Pi/minㆍmg였으며, 비운동구에서 0.34μmol Pi/minㆍmg으로 운동시킨 참돔의 육에서 ATPase 활성이 약간 높은 것으로 나타났다. 운동사육시킨 참돔은 운동기간이 증가할수록 ATPase활성이 증가하여 운동 20일에는 0.42μmo1 Pi/minㆍmg의 함량을 나타내었으나, 비운동사육에서는 큰 차이를 보이지 않고 있다. 근원섬유의 Ca/sup 2+/가 첨가되지 않았을때의 Mg/sup 2+/-ATPase 활성을 나타내었다. 운동시키전에는 각각 0.10μmol Pi/minㆍmg, 0.14μmol Pi/minㆍmg 나타나 비운동구가 운동구에 비하여 Mg/sup 2+/(-Ca/sup 2+/)-ATPase 활성이 높았으며 사육기간동안 활성이 서서히 감소하는 경향을 나타내었다. Ca/sup 2+/-ATPase의 활성 또한 사육기간 동안 큰 변화를 보이지 않았다. 그러므로, 참돔의 운동여부에 따라서 ATPase활성을 살펴본 결과, 운동기간에 따른 약간의 차이는 보이고 있으나, 품질지표로써 사용하기가 어렵다고 판단된다. 효소적인 방법을 이용한 양식산 활어의 품질지표로써 ATP 관련화합물, ATPase 활성은 운동여부, 크기에 따라 객관적인 지표로써의 사용이 어려우므로, 건강도를 측정할 수 있는 AEC의 수치를 적용하는 것이 적합하다고 판단된다. 즉, 양식산 참돔의 AEC 수치가 1∼0.9의 범위에서는 건강한 활어로 상급의 등급을, 0.8∼0.7의 AEC 수치는 중급의 등급으로, 그 이하의 수치에서는 하급으로 적용가능 하다고 판단된다.

• The Journal of Internal Korean Medicine
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• v.19 no.1
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• pp.431-441
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• 1998

This study was undertaken to determine whether Buthus exract(BTE) affects Na^+-K^+-ATPase$ activity of nervous tissues. The enzym activity was measured in synaptosomal fraction prepared from rabbit brain cortex. Na^+-K^+-ATPase$ activity was inhibited by BTE over concentration range of 0.05-0.5% in a dose-dependent manner. The enzyme activity was increased by an increase in $Na^+$ concentration from 5 to 100mM, $K^+$ concentration from 0.5 to 10mM, and $Mg^{2+}$ concentration from 0.2 to 5mM. These changes in ion concentrations did not produce any effect on the inhibitory effect of BTE on $Na^+-K^+-ATPase$ activity. An increase in ATP concentration from 0.1 to 3mM caused an increase in the enzyme activity. The inhibition of the enzyme activity by BTE were not different between two ATP concentrations. A sulfhydryl group protector DTT prevented PCMB-induced inhibition of $Na^+-K^+-ATPase$ activity, but the BTE-induced inhibition was not altered by DTT. The inhibition of enzyme activity by combination of ouabain and BTE was not different from that by Buthus alone. These results suggest that Buthus exerts inhibitory effect on $Na^+-K^+-ATPase$ activity in cerebral synaptosomes, and the action mechansim is similar to that of ouabain.

• The Korean Journal of Mycology
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• v.15 no.4
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• pp.224-230
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• 1987

Effects Of organic compound, photosensitizer and $K^+$ ion influx. On the light-induced ATPase of mitochondria in L. edodes purified by linear sucrose density gradient centrifugation were studied. The mitochondrial ATPase activity was investigated by various wavelength illumination at dark state. The mitochondrial ATPase was activated 139% and 128% by 10m mol dithiothreitol and 0.1m mol quinacrine, respectively. This enzyme also was activated 36% by 0.1m mol phenazine methosulfate as photosensitizer. But, 100 mg oligomycin and 1m mol phlorizin inhibited activity of enzyme to 48% and 45%, respectively. Its optimum wavelength was 690 nm on the effect of $K^+$ ion influx, its optimum pH and temperature were found to be 7.2 and $55^{\circ}C$.

• Korean Journal of Food Science and Technology
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• v.25 no.2
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• pp.185-190
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• 1993

Tilapia cultured in fresh water were acclimated in sea water with daily increase of $5%_{\circ}$ of salinity and it was completely terminated at the 7th day (0 week). Each three tilapia acclimated were examined weekly based from 0 week to elucidate changes of chloride cells in gill, mineral contents and physical properties in muscle and biochemical characteristics in myofibrils. Chloride cells existed in gills were gradually developed in number and size by acclimation to sea water and became to almost constant state at the third week. Shearing value, compressing strength and content of minerals such as Mg, Na and K in muscle were showed remarkable increase by acclimation to sea water in comparison to those of muscle from tilapia reared in fresh water. Myofibrillar $Mg^{2+}-,\;Ca^{2+}-$ and $K^+(EDTA)-ATPase$ activities of tilapia acclimated in sea water also increased showing significant statistic difference (p<0.01) from those of tilapia reared in fresh water However. thermostability of myofibrils was dropped by acclimation to sea water. The increase of shearing value and compressing strength in the muscle of tilapia by acclimation to sea water would be attributed to the increase of myofibrillar ATPase activities which act to accelerate the decomposition rate of ATP. Therefore, it is suggested that this phenomenon associated with muscle contraction could be brought an improvement of texture of tilapia acclimated in sea water.

• Korean Journal of Environmental Biology
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• v.32 no.4
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• pp.344-352
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• 2014

The aim of the present study was to assess the effects of ammonia on physiological responses in Korean rockfish (Sebastes schlegeli). Interestingly, no mortality were observed when the specimens ($301.1{\pm}8.0g$) were exposed to five levels of un-ionized ammonia ($NH_3$) (control, 1, 2, 4, $8mg\;L^{-1}$) for 3 hours. Furthermore, a significantly higher increase in gill $Na^+/K^+$-ATPase (NKA) pump activity with was detected due to the ammonia exposure. The activity of the fishes were found to be 4 and $8mg\;L^{-1}$ $NH_3$, which was significantly high compared to normal $1mg\;L^{-1}$ $NH_3$. Although ammonia exposure had no effect on plasma $Cl^-$, exposure to both 4 and $8mg\;L^{-1}$ $NH_3$, however it led to increase in the plasma $Na^+$, $K^+$ and osmolality levels. Also, prolong ammonia exposure cause increase of plasma cortisol and glucose levels. The increase in glucose was accompanied by an increase in cortisol. The fish exposed to 4 and $8mg\;L^{-1}$ $NH_3$ showed significantly higher hematocrit than control group than those exposed to 1 and $2mg\;L^{-1}$ $NH_3$. The intensity of cell damage increased with the increase concentration and exposure to ammonia. Furthermore, hyperplasia, separation and epithelial necrosis were also observed in gill tissues. Taken together, the results showed that direction of changes to the investigated parameters can be used to determine the physiological responses of Korean rockfish to ammonia.

• Toxicological Research
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• v.34 no.2
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• pp.143-150
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• 2018

It has been demonstrated that vanadate causes nephrotoxicity. Vanadate inhibits renal sodium potassium adenosine triphosphatase (Na, K-ATPase) activity and this is more pronounced in injured renal tissues. Cardiac cyclic adenosine monophosphate (cAMP) is enhanced by vanadate, while increased cAMP suppresses Na, K-ATPase action in renal tubular cells. There are no in vivo data collectively demonstrating the effect of vanadate on renal cAMP levels; on the abundance of the alpha 1 isoform (${\alpha}_1$) of the Na, K-ATPase protein or its cellular localization; or on renal tissue injury. In this study, rats received a normal saline solution or vanadate (5 mg/kg BW) by intraperitoneal injection for 10 days. Levels of vanadium, cAMP, and malondialdehyde (MDA), a marker of lipid peroxidation were measured in renal tissues. Protein abundance and the localization of renal ${\alpha}_1-Na$, K-ATPase was determined by Western blot and immunohistochemistry, respectively. Renal tissue injury was examined by histological evaluation and renal function was assessed by blood biochemical parameters. Rats treated with vanadate had markedly increased vanadium levels in their plasma, urine, and renal tissues. Vanadate significantly induced renal cAMP and MDA accumulation, whereas the protein level of ${\alpha}_1-Na$, K-ATPase was suppressed. Vanadate caused renal damage, azotemia, hypokalemia, and hypophosphatemia. Fractional excretions of all studied electrolytes were increased with vanadate administration. These in vivo findings demonstrate that vanadate might suppress renal ${\alpha}_1-Na$, K-ATPase protein functionally by enhancing cAMP and structurally by augmenting lipid peroxidation.

• The Korean Journal of Pharmacology
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• v.16 no.1 s.26
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• pp.51-56
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• 1980

Phytolaccae Radix (PR), Brunella Herba (BH), Akebiae Lignum (AL) and Atractylis Rhizoma (AR) are some of the diuretic agents used in Chinese medicine and folk remedy. Water or methanol extracts of them (100mg/kg) were intravenously injected to rabbits in order to re-evaluate the effects on renal function. PR water extract elicited moderate diuresis while water extracts of BH, AL and methanol extract of AR had antidiuretic effects. Influence of PR on renal hemodynamics and $Na^+-K^+$-ATPase activity in rabbit kidney were observed in vivo and in vitro. The results were as follows: 1) Clearances of inulin and p-aminohippuric acid increased significantly after 15 minutes following the administration of PR water extract, but Na+ reabsorption rate was not changed. 2) The increase of $Na^+-K^+$-ATPase activity in renal cortex, outer and inner medulla was observed at 15 minutes after PR water fraction was given intravenously, and the change was most prominent in cortical area. 3) More than 50% of decrease in $Na^+-K^+$-ATPase activity in renal tissues was observed with PR water fraction $(10^{-2}g/ml)$ in vitro experiments. However, the inhibition of $Na^+-K^+$-ATPase activity was reversed with lower concentrations $(10^{-4}g/ml,\;10^{-6}g/ml)$ of PR water fraction in outer and inner medullary zone. These results suggest the diuretic effect of PR is due to improved renal hemodynamics, and contradictory reults concerning $Na^+-K^+$-ATPase activity require further investigation.

• The Korean Journal of Physiology
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• v.17 no.2
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• pp.125-133
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• 1983

Red cell glycolytic intermediates, metabolites and metabolic ratios were studied. Glycolytic intermediates were measured in neutralized perchloric acid extracts of red cell suspensions after 3 hr incubation at $37^{\circ}C$ in the presence and absence of saponin. Adenosine triphosphate(ATP), adenosine diphosphate(ADP), pyruvate and lactate were measured by enzymatic procedures involving stoichiometric oxidation or reduction of a pyridine nucleotide. Glucose was determined using glucose oxidase after zinc hydroxide extraction. The redox state was calculated from the lactate dehydrogenase equilibrium. Adenosine triphosphatase activity(ATPase) was measured by determining the amount of phosphate released from ATP by washed erythrocyte membranes(ghost) during 20 min. incubation. Both total hydrolysis and the amount of hydrolysis that occured in the presence of ouabain were measured. The second measurement yields Mg-ATPase and represents nonspecific ATPase activity of the membranes. The difference between total and Mg-ATPase activity can be attributed to Na-K-ATPase. For the measurement of sodium fluxes, human erythrocytes were preincubated in $^{22}Na$ for 3 hr at $37^{\circ}C$, washed and suspended in a tracer-free medium. The amount of $^{22}Na$ transported out of cells at any time was determined by analysis of supernatant samples taken at various time after addition of the labeled cells to isotope-free medium. The cells and medium were separated and the radioactivity appearing in the medium was measured. From the total radioactivity in the suspension and the radioactivity appearing in the medium at known time, the rate constant for sodium release was computed. The results are summarized as follows: 1) ATP and ATP/ADP were found to increase at every concentration of saponin tested whereas ADP declined at every cone. of saponin. The increase in pyruvate and lactate were observed at every cone, of saponin and thus $NAD^+/NADH$ computed from pyruvate/lactate also increased. Glucose utilization was stimulated by saponin. 2) $Na^+-K^+-ATPase$ activities showed a biphasic response to saponin, first increasing in lower concentration and then decreasing in higher concentration of saponin. 3) The efflux of sodium was significantly increased by saponin in the range of 5 to 10 mg%. The stimulatory effect of saponin on the rate constants for active(ouabain-sensitive) sodium efflux was inhibited by addition of ouabain.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.6
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• pp.609-616
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• 2017

Ardipusilloside-I is a natural triterpenoid saponin, which was isolated from Ardisia pusilla A. DC. The aim of the study was to evaluate the stimulation of ardipusilloside-I on gastrointestinal motility in vitro and in vivo. The experiment of smooth muscle contraction directly monitored the contractions of the isolated jejunal segment (IJS) in different contractile states, and the effects of ardipusilloside-I on myosin were measured in the presence of $Ca^{2+}$-calmodulin using the activities of 20 kDa myosin light chain ($MLC_{20}$) phosphorylation and myosin $Mg^{2+}$-ATPase. The effects of ardipusilloside-I on gastro emptying and intestinal transit in constipation-predominant rats were observed, and the MLCK expression in jejuna of constipated rats was determined by western blot. The results showed that, ardipusilloside-I increased the contractility of IJS in a dose-dependent manner and reversed the low contractile state (LCS) of IJS induced by low $Ca^{2+}$, adrenaline, and atropine respectively. There were synergistic effects on contractivity of IJS between ardipusilloside-I and ACh, high $Ca^{2+}$, and histamine, respectively. Ardipusilloside-I could stimulate the phosphorylation of $MLC_{20}$ and $Mg^{2+}$-ATPase activities of $Ca^{2+}$- dependent phosphorylated myosin. Ardipusilloside-I also stimulated the gastric emptying and intestinal transit in normal and constipated rats in vivo, respectively, and increased the MLCK expression in the jejuna of constipation-predominant rats. Briefly, the findings demonstrated that ardipusilloside-I could effectively excite gastrointestinal motility in vitro and in vivo.