The trypanocidal drug suramin, an impermeant polyanion, has been shown to be a powerful inhibitor of the calcium uptake and calcium-stimulated ATPase activity of sarcoplasmic reticulum (Fortes et al., 1974). In view of this finding, an attempt was made to investigate the effect of suramin on $Ca^{++}$ transport in resealed red cells and on $Ca^{++}$-activated ATPase in red blood cell membrane fragments (RBCMF). The results obtained are summarized as follows. 1. $Ca^{++}$ outflux from the resealed RBC was inhibited by suramin and the inhibitory action of suramin is proportional to the concentration of drug added inside the RBC preparation. When suramin is added both inside and outside the RBC preparation simultaneously, the magnitude of the inhibitory effect was more pronounced, suggesting that suramin inhibits both active $Ca^{++}-^{45}Ca$ exchange diffusion across the RBC membrane. 2. Suramin inhibits the $Ca^{++}$-activated ATPase of the RBCMF and the effect of inhibition by the drug was also concentration dependent. From the above results, it may be concluded that suramin inhibits $Ca^{++}$ transport across RBC membrane by inhibiting $Ca^{++}$-activated ATPase activity which has been known to be linked with active $Ca^{++}$ transport.
Jung, Dai Hyun;Lee, Cheol Joo;Suh, Chang Kook;You, Hye Jin;Kim, Do Han
Molecules and Cells
/
v.20
no.1
/
pp.51-56
/
2005
Excitation-contraction coupling (ECC) proteins in the human heart were characterized using human atrial tissues from different age groups. The samples were classified into one infant group (Group A: 0.2-7 years old) and three adult groups (Group B: 21-30; Group C: 41-49; Group D: 60-66). Whole homogenates (WH) of atrial tissues were assayed for ligand binding, $^{45}Ca^{2+}$ uptake and content of ECC proteins by Western blotting. Equilibrium [$^3H$]ryanodine binding to characterize the ryanodine receptor (RyR) of the sarcoplasmic reticulum (SR) showed that the maximal [$^3H$]ryanodine binding ($B_{max}$) to RyR was similar in all the age groups, but the dissociation constant ($k_d$) of ryanodine was higher in the infant group than the adult groups. Oxalate-supported $^{45}Ca^{2+}$ uptake into the SR, a function of the SR SERCA2a activity, was lower in the infant group than in the adult groups. Similarly, [$^3H$]PN200-110 binding, an index of dihydropyridine receptor (DHPR) density, was lower in the infant group. Expression of calsequestrin and triadin assessed by Western blotting was similar in the infant and adult groups, but junctin expression was considerably higher in the adult groups. These differences in key ECC proteins could underlie the different $Ca^{2+}$ handling properties and contractility of infant hearts.
This study was undertaken to evaluate the effect of Sunghyangchungi-san (SHCS) on the oxidant-induced contraction and lipid peroxidation in rabbit carotid artery. Vascular rings isolated from rabbit carotid artery were exposed to t-butyl hydroperoxide (t-BHP), an extrinsic oxidant, and the effect of SHCS on the changes of vascular tension and lipid peroxidation induced by t-BHP was determined. t- BHP induced a slowly developing and sustained contraction of the arterial rings. SHCS effectively relaxed the arterial rings that were pre-contracted by t-BHP. The responses to SHCS were partially dose-dependent at concentrations lower than 0.5 mg/ml. When SHCS was applied prior to the exposure to t-BHP, it inhibited the t-BHP-induced contraction as well. t- BHP increased lipid peroxidation in a dose-dependent manner. SHCS as well as well-known anti-oxidants GSH and DPPD reduced significantly lipid peroxidation induced by t-BHP. SHCS partially blocked the increase in $^{45}Ca$ uptake induced by t-BHP. In contrast to SHCS, anti-oxidants GSH and DPPD failed to inhibit significantly the t- BHP-induced contraction or $^{45}Ca$ uptake. From the above results, it is suggested that SHCS relaxed t-BHP-induced contraction of rabbit carotid artery independently of its anti-oxidant action, and inhibition of $Ca^{2+}$ influx may contribute to the underlying mechanism.
A pot experiment was carried out to investigate the effect of Cd and Zn added to soil on the growth, peroxidase activity, chlorophyll content, inorganic components, and interaction between Cd and Zn in corn plant. The results obtained are as follows. Both Cd and Zn reduced dry weight and height of corn plant. Peroxidase activity of leaves increased in higher Cd content of shoot but adverse trend was shown with increasing Zn content of shoots. Chlorophyll contents were significantly decreased with increase of both Cd and Zn content of shoots. Higher soil Cd levels increased N content but decreased P, K, Ca, Mg and $SiO_2$ content in plant. Wherease higher Zn levels in soil increased P and $SiO_2$ content but decreased N, Ca, Mg content in plant. The total Cd and Zn uptake of shoot increased with time but the Cd and Zn content of shoot showed no tendency. Cadmium uptake by Zn application was reduced in higher Zn levels with time while Zn uptake by Cd application showed adverse trend.
Park, Hae-Kun;Jeon, Byeong-Hwa;Kim, Se-Hoon;Kim, Hoe-Suk;Chang, Seok-Jong
The Korean Journal of Physiology
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v.28
no.2
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pp.181-190
/
1994
Endothelium-derived relaxing factor (EDRF) activates guanylate cyclase which mediates the formation of cGMP from GTP in vascular smooth muscle. It is well known that endothelium-dependent relaxation is impaired in spontaneously hypertensive rats (SHR). However, it is still unknown whether the impaired endothelium-dependent relaxation in SHR results from the reduced release of EDRF or from the decrease of vascular response to EDRF. We investigated the effects of cGMP on the contractility and Ca movement in the aorta of SHR and Wistar-Kyoto rats (WKY). The amplitude of the endothelium-dependent relaxation to actylcholine (ACh) was significantly less in SHR than in WKY. L-arginine $(10^{-3}M)$ did not increase endothelium-dependent relaxation in both strains. Sodium nitroprusside (SNP), an activator of guanylate cyclase, relaxed the 40 mM $K^+-induced$ contraction in a dose-dependent manner $(10^{-10}{\sim}10^{-6}\;M)$ in the endothelium-rubbed aortic strips of both strains. However, there was no significant difference in these relaxations between WKY and SHR. 8-bromo-cyclic guanosine monophosphate (8-Br-cGMP), a cell membrane-permeable derivative of cGMP relaxed the 40 mM $K^+-induced$ contraction in a dose-dependent manner $(10^{-6}{\sim}10^{-4}\;M)$ in the endothelium-rubbed aortic strips of both strains. Also norepinephrine $(10^{-6}\;M)-induced$ contractions in normal and Ca-free Tyrode's solution were suppressed by the pretreatment with 8-Br-cGMP $(10^{-4}\;M)$ in either strain. However, the amplitudes of suppression induced by 8-Br-cGMP were greater in SHR than that in WKY. Basal $^{45}Ca$ uptake and 40mM $K^+-stimulated\;^{45}Ca$ uptake were not suppressed by pretreatment with 8-Br-cGMP $(10^{-4}\;M)$ in single aortic smooth muscle cells of both SHR and WKY. From the above results, it is suggested that cGMP decreases Ca sensitivity in vascular smooth muscle cells and that the impaired endothelium-dependent relaxation in the aortic strips of SHR is not the result of a reduced vascular response to EDRF.
Tc-99m MIBI, a lipophilic cation, was reported as a useful agent for localization of lung cancer. The effect of radiation therapy on the uptake of Tc-99m MIBI in lung cancer, however, was not well evaluated. The aim of the present study was to elucidate the usefulness of Tc-99m MIBI SPECT in the localization and the assessment of radiotherapy in non-small cell lung cancer. Twenty patients(19 males and 1 female, mean age 59, 16 squamous cell ca and 4 adenoca) were studied with Tc-99m MIBI SPECT before radiation therapy. Eleven patients(10 males and 1 female, mean age 59, 8 squamous cell ca and 3 adenoca) were repeated the study 1 month after the completion of radiation therapy(mean dose 6453cGy). All patients showed positive uptakes of Tc-99m MIBI in their tumors. One patient showed a hot uptake in atelectatic area. There was no difference of Tc-99m MIBI uptakes between squamous cell ca and adenoca either on planar or tomographic images. Tc-99m MIBI uptake ratios of squamous cell ca and adenoca were $1.50{\pm}0.16$ and $1.45{\pm}0.15$ on planar images, and $2.73{\pm}0.46$ and $2.54{\pm}0.37$ on tomographic images, respectively. The concordance between radiological change(chest x-ray and CT) and change of Tc-99m MIBI uptakes was 9/11 (81.8% ). In conclusion, Tc-99m MIBI SPECT was useful in the localization of tumor and the assessment of radiation therapy in non-small cell lung cancer.
Recently, we reported that GS 389 has vasodilating action without cardiac inotropic action (Chang et al., Can. J. Physiol. Pharmacol. 72, 327-334, 1994). However the mechanism of action of GS 389 has not been thoroughly evaluated. In the present study, we performed functional study of GS 389 in rat trachealis, thoracic aorta, pig coronary artery by isometric tension and in human platelets by aggregation experiments. We also tested if GS 389 influences on $Ca^{2+}$movement and inositol phosphate metabolism. In rat trachealis, GS 389 concentration-dependently relaxed carbachol (0.1 $\mu$M)- and high $K^{+}$(65.4 mM)-induced contraction with p$IC_{50}$/ of 4.43$\pm$ 0.19 and 4.11$\pm$0.12, respectively. In $Ca^{2+}$-free media, GS 389 inhibited carbachol-induced phasic contraction. In rat thoracic aorta, GS 389 inhibited $^{45}$ Ca uptake due to norepinephrine and high $K^{+}$, indicating that GS 389 has direct inhibitory action of $Ca^{2+}$movement. Furthermore, GS 389 competitively inhibited U46619-induced contraction in rat thoracic aorta and pig coronary artery with K, values of 5.23$\pm$0.12 and 5.56$\pm$0.14, respectively, and inhibited U 46619-induced phosphatidylinositide (PI) turnover in rat aorta. GS 389 also concentration-dependently inhibited the human platelet aggregation against U 46619 with p$IC_{50}$/ 5.66$\pm$0.02. These results indicate that GS 389 has thromboxane $A_2$ antagonistic action in vascular and platelets as well as direct action on $Ca^{2+}$ movement, which may account, at least in part, for relaxing action of rat trachealis. trachealis.
Sarcolemmal membrane fraction from canine ventricle was isolated from the discarded pellet after the first homogenization in the isolation procedure of sarcoplasmic reticulum (Method 1) and the protein yield, purity, and sidedness of this preparation were compared to those of sarcolemmal fraction prepared by method of Lee et al. (Method 2) and a slight modification of original protocol of Jones et al. (Method 3). Method 1 differed from Method 2 essentially only in that vigorous homogenization was carried out by omnimixer and homogenization medium containing 30mM Tris-maleate was used in the first step. The sarcolemmal fraction was enriched from 45 to 50 and 29-fold in [$^3H$] ouabain, [$^3H$] DHA, [$^3H$] QNB binding and $Na^+$, $K^+$-ATPase activity, respectively, compared to homogenate. Total $Na^+$, $K^+$-ATPase activity of highly sarcolemma enriched fraction was 144.6$\pm$16.4$\mu\textrm{mol}$ Pi/mg protein/hr, which was about 85%, of total ATPase activity, and the yield of the preparation was 15.7 mg protein per 100g of starting ventricular tissue. The sarcolemmal preparation supported $^{45}Ca^{2+}$-uptake in the presence of ATP but this uptake was not dependent on oxalate. Sarcolemmal $Na^+$, $K^+$-ATPase activity and detectable [$^3H$] ouabain binding were increased about 32% and 35%, respectively, by pretreatment of sarcolemmal fraction with optimal concentration of sodium dodecylsulfate (0.3-0.4mg/mg protein), suggesting that this preparation contained about 24% of sealed rightside-out vesicles, 26% of sealed inside-out vesicles, and 5001o of freely permeable (leaky) form. This procedure showed the highest protein yield and leaky population, compared to Method 2 and 3. On the other hand, sarcolemmal fraction prepared by Method 2 and 3 showed low value in protein yield but comtained high population of inside-out (46%) and rightside-out (49%) vesicles, respectively, compared to present procedure (Method 1). The results indicate that vigorous homogenization decreases the population of sealed sarcolemmal vesicles but increases the sarcolemmal protein yield per gram tissue and that this procedure is available for further purification of sarcolemmal fraction and for the receptor binding study of sarcolemma.
Objective : This study was undertaken to evaluate the effect of Sunghyangchungisan (SHCS) on the regulation of vascular tone and $Ca^{2+}$ metabolism in arterial tissues. Vascular rings isolated from rabbit carotid artery were myographed isometrically in isolated organ baths and the effect of SHCS on contractile activities, endothelial function and $Ca^{2+}$ metabolism were determined. Methods : In phentobarbital sodium-anesthetized rabbits, SHCS administered through ear vein (100 mg/Kg body wt.) or intragastric dwelling tube (300 mg/Kg body wt.) attenuated phenylephrine (PE, 10 ${\mu}g$/Kg, i.v.)-induced increases in both systolic and diastolic cartoid arterial blood pressure. Results : In experiments with isolated arterial strips, SHCS relaxed arterial rings which were pre-contracted by phenylephrine (PE, 1 ${\mu}M$). The responses to SHCS were partially dose-dependent at concentrations lower than 0.5 mg/ml. When SHCS was applied prior to the exposure to PE, it inhibited the PE-induced contraction by a similar magnitude which was comparable to the relaxation of pre-contracted arterial rings. Washout of SHCS after observing its relaxant effect resulted in a full recovery of PE-induced contractions, indicating that the action mechanism is reversible. The observation that SHCS did not change the $ED_{50)$ of PE oh its dose-response curve ruled out the possible interaction of SHCS with ${\alpha}$-receptors. The relaxant effect of SHCS was not affected by removal of endothelium or a nitric oxide synthase inhibitor, L-NAME. Methylene blue, an inhibitor of the soluble guanylate cyclase, did not affect the relaxant effect of SHCS. These results suggest that the action of SHCS is not mediated by the endothelium nor soluble guanylate cyclase. Constant cGMP production determined in arterial strips in the presence or absence of SHCS is consistent with this conclusion. When contraction was induced by additive application of $Ca^{2+}$ in arterial rings which were pre-depolarized by high $K^+$ in a $Ca^{2+}$-free solution, the relaxant effect of SHCS was attenuated by increasing the $Ca^{2+}$ concentration. SHCS, when applied to the arterial rings pre-contracted by PE and then relaxed by nifedipine, a $Ca^{2+}$ channel blocker, did not show additive relaxation. SHCS partially blocked $Ca^{2+}$ influx stimulated by PE and high $K^+$ which was determined by 5-min ^{45}Ca$ uptake, while it did not affect $Ca^{2+}$ efflux. Conclusions : From above results, it is suggested that SHCS relax PE-induced contraction of rabbit carotid artery in an endothelium independent manner, andinhibition of $Ca^{2+}$ influx may contribute to the underling mechanism.
To promote the practical use of livestock manure compost (LC) for paddy rice cultivation, the fertilization efficiency of nutrients in LCs was investigated compared to that of chemical fertilizer. This experiment was conducted at rice field in Hwaseong, Korea, with 6 treatments by each of 3 kinds of tested LCs, cattle manure compost (CaC), swine manure compost (SwC) and chicken manure compost (ChC). The treatments consisted of 3 application levels of LCs and 3 chemical fertilizer treatments having the same application levels with LCs. $NH_4$-N content in soil became higher according to the increase in the urea application rate, while it became lower in LC plots than in urea plots, and statistically had no significant difference among LC plots. There was a close relationship between phosphate fertilization rate and the increment of soil available phosphate content after experiment resulting y = 0.1788x - 6.169 ($R^2=0.9425$) when applied fused superphosphate fertilizer, and y = 0.0662x - 2.689 ($R^2=0.9315$) when applied LC at the equivalent rates to phosphate input (x: phosphate application rate, kg $ha^{-1}$, y: increment in soil available phosphate content, mg $kg^{-1}$). And from these two equations, the correlation on the phosphate application rate between fused superphosphate fertilizer and LC could be obtained as y = 2.7056x - 52.492 (x: $P_2O_5$ application rate of fused superphosphate, kg $ha^{-1}$, y: $P_2O_5$ application rate of LC, kg $ha^{-1}$). Plant height, number of tillers, nutrients uptake by rice, and rice yield showed higher levels in N 100% and N 150% application plots of chemical fertilizers, while every LC plots exhibited lower values and no significant difference among them. Relative nitrogen fertilization efficiencies of LCs compared to urea was 12.3% for CaC, 8.8 for SwC and 24.6 for ChC, respectively.
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