In this study, before and after sunset carbon dioxide concentration and air temperature were observed in two points of atmosphere (lower observation point of the GL + 0.1 m, the upper observation point of GL + 1.0 m) on the foreshore at located in Suncheon Bay and their variations were analyzed. Observation was performed on the foreshore on 2~4 August 2010. Instrument (VAISALA, GMP343) was set two hours before sunset and then observation was made continuously for six hours. In three days, observed carbon dioxide concentration was 375~419 ppm, and the air temperature was in the range of $28.7{\sim}32.5^{\circ}C$. The average concentration of carbon dioxide was 388~399 ppm in the upper observation point and 386~396 ppm in the lower observation point. It was higher in the upper observation point and its fluctuations were similar in two observation points. Correlation coefficients between carbon dioxide concentration and air temperature in the upper observation point were in the range of -0.64~-0.88, and were calculated -0.65 to -0.90 in the lower observation point. For the carbon dioxide concentration, correlation coefficients between the upper part and the lower part were very high as 0.98 in three times. For the air temperature, correlation coefficients between the upper part and the lower part were very high as 0.97 and 0.99. In the same observation time, the slope of the linear regression function as carbon dioxide concentration in the lower observation point for the upper observation point was in the range of 0.97~1.01. Carbon dioxide concentration was slightly higher in the upper observation point. Because carbon dioxide in the lower observation point was closer on the surface of the foreshore and absorbed from atmosphere to the foreshore. In this study, it was showed that the vertical variation of carbon dioxide concentration was insignificant in the several meter scale of atmosphere on the surface of the foreshore.
An enzyme that hydrolyzes flavin-adenine dinucleotide (FAD) was found to be present in rat liver lysosomal membrane prepared from Triton WR-1339 filled lysosomes (tritosomes) purified by flotation on sucrose. This FAD phosphohydrolase (FADase) exhibited optimal activity at pH 8.5 and had an apparent Km of approximately 3.3 mM. The activity was decreased 50~70% by dialysis against EDTA and this was restored by $Zn^{2+}$, $Mg^{+2}$, $Hg^{+2}$, and $Ca^{+2}$ ions inhibited the enzyme, but $F^-$ and molybdate had no effect. The enzyme was also inhibited by p-chloromercuribenzoate (pCMB), reduced glutathione and other thiols, cyanide, and ascorbate. The presence of ATP, ADP, AMP. ${\alpha}-{\beta}-methylene$ ATP, AMP-p-nitrophenyl phosphate (PNP), GMP, and coenzyme A (CoA) decreased the activity on FAD, but pyrimidine nucleotides, adenosine, adenine, or $NAD^+$ were without effect. Phosphate stimulated the activity slightly. FAD phosphohydrolase activity was separated from ATPase and inorganic pyrophosphatase activities by solubilization with detergents and polyacrylamide gel electrophoresis and by linear sucrose density gradient centrifugation suggesting that the enzyme is different from ATPase, inorganic pyrophosphatase, and soluble lysosomal FAD pyrophosphatase. Paper chromatography showed that FAD was hydrolyzed to flavin mononucleotide (FMN) and AMP which were further hydrolyzed to riboflavin and AMP by phosphatases known to be present in lysosomal membranes. Incubation of the intact Iysosomes with pronase showed that the active site of FAD phosphohydrolase must be oriented to the cytosol. The FAD hydrolyzing activity was detected in Golgi, microsome, and plasma membrane, but not in mitochondria or soluble lysosomal preparations.
Objectives: Radix Ginseng has been used for thousands of years to treat a wide variety of diseases. Radix ginseng has also been used as a traditional medicine for boosting Qi energy and tonifying the spleen and lungs. Traditionally, its effect could be obtained orally. Nowadays, a new method, the injection of herbal medicine, is being used. This study was performed to investigate the single-dose intravenous toxicity of water-soluble ginseng pharmacopuncture (WSGP) in Sprague-Dawley (SD) rats. Methods: All experiments were carried out at Biotoxtech, an institute authorized to perform non-clinical studies under the regulation of Good Laboratory Practice (GLP). At the age of six weeks, 40 SD rats, 20 male rats and 20 female rats, were allocated into one of 4 groups according to the dosages they would receive. The WSGP was prepared in the Korean Pharmacopuncture Institute under the regulation of Korea-Good Manufacturing Practice (K-GMP). Dosages of WSGP were 0.1, 0.5 and 1.0 mL/animal for the experimental groups, and normal saline was administered to the control group. The rat's general conditions and body weights, the results of their hematological and biochemistry tests, and their necropsy and histopathological findings were investigated to identify the toxicological effect of WSGP injected intravenously. The effect was examined for 14 days after the WSGP injection. This study was performed under the approval of the Institutional Animal Ethics Committee of Biotoxtech. Results: No deaths were found in this single-dose toxicity test on the intravenous injection of WSGP, and no significant changes in the rat's general conditions and body weights, the results on their hematological and biochemistry test, and their necropsy findings were observed during the test. The local area of the injection site showed minial change. The lethal dose was assumed to be over 1.0 mL/animal in both sexes. Conclusion: These results indicate that WSGP is safe at dosages up to 1 mL/animal.
BACKGROUND/OBJECTIVES: This study evaluated the effects and molecular mechanisms of the Schisandra chinensis fruit extract (SC) and its major compound gomisin A (GA), on the contractility of rabbit penile corpus cavernosum smooth muscle (PCCSM). MATERIALS/METHODS: PCCSM was exposed to SC or GA after appropriate pretreatment with nitric oxide synthase (NOS) blocker, guanylate cyclase blocker, adenylyl cyclase blocker or protein kinase A blocker. Subsequently, we evaluated the cyclic nucleotide in the perfusate by radioimmunoassay, protein expression level of neuronal NOS (nNOS) and endothelial NOS (eNOS) by western blot, and the interaction of SC or GA with udenafil and rolipram. RESULTS: Both SC and GA induce PCCSM relaxations in a concentration-dependent manner. Pretreatment with NOS blocker, guanylate cyclase blocker, adenylyl cyclase blocker or protein kinase A blocker result in significantly decreased relaxation. SC and GA also induce the levels of cyclic nucleotide in the perfusate in a concentration-dependent manner. Perfusion with GA also showed significantly higher levels of eNOS protein. Furthermore, the udenafil and rolipram induced relaxations of PCCSM were enhanced after exposure to SC and GA. Our results indicate that SC and GA induce the relaxation of PCCSM via the nitric oxide (NO)-cGMP and cAMP signaling pathways. CONCLUSIONS: The SC and GA are potential alternative treatments for men who want to consume natural products to ameliorate erectile function, or who do not respond to the commercially available medicines.
Glutamate uptake inhibitor, L-trans-pyrrolidine-2, 4-dicarboxylate (PDC, $20{\mu}M$) elevated basal and N-methyl-D-aspartate (NMDA, $100{\mu}M$)-induced extracellular glutamate accumulation, while it did not augment kainate $100{\mu}M$-induced glutamate accumulation in cultured cerebellar granule neurons. However, pretreatment with PDC for 1 h significantly reduced NMDA-induced glutamate accumulation, but did not affect kainate-induced response. Pretreatment with glutamate $(5{\mu}M)$ for 1 h also reduced NMDA-induced glutamate accumulation, but did not kainate-induced response. Upon a brief application (3-10 min), PDC did neither induce elevation of intracellular calcium concentration $([Ca^{2+}]_i)$ nor modulate NMDA-indLiced $[Ca^{2+}]_1$ elevation. Pretreatment with PDC for 1 h reduced NMDA-induced $[Ca^{2+}]_1$ elevation, but it did not reduce kainate-induced $[Ca^{2+}]_1$ elevation. These results suggest that glutamate concentration in synaptic clefts of neurana cells is increased by prolonged exposure (1 h) of the cells to PDC, and the accumulated glutamate subsequently induces selective desensitization of NMDA receptor.
The aim of this study is to investigate the effect of anodizing surface to osseointegration of implant by using of resonance frequency analysis (RFA), quantitative and qualitative assessment of an anodically modified implant type with regard to osseous healing qualities. A total of 96 screw-shaped implants were prepared for this study. 72 implants were prepared by electrochemical oxidation with different ways. 24 (group 1 SP) were prepared at galvanostatic mode in 0.25M sulfuric acid and phosphoric acid. 24 (group 2GC) were prepared at galvanostatic mode in calcium glycerophosphate and calcium acetate and 24 (group 3 CMP (Calcium Metaphosphate) Coating were prepared at galvanostatic mode in 0.25M sulfuric acid and phosphoric acid followed by CMP coating. Rest of 24 (control group were as a control group of RBM surface. Bone tissue responses were evaluated by resonance frequency analysis (RFA) that were undertaken at 2, 4 and 6 weeks after implant placement in the mandible of mini-pig. Group 1 SP (anodized with sulfuric acid and phosphoric acid implants) demonstrated slightly stronger bone responses than control Group RBM. Group 2 GC (anodized surface with calcium glycerophosphate and calcium acetate implants) demonstrated no difference which were compared with control group. Group 3 GMP (anodized and CMP coated implants) demonstrated slightly stronger and faster bone responses than any other implants. But, all observation result of RF A showed no significant differences between experimental groups with various surface type. Histomorphometric evaluation demonstrated significantly higher bone-to-implant contact for group 2 GC. Significantly more bone formation was found inside threaded area for group 2 GC. It was concluded that group 2 GC (anodized surface with calcium glycerophosphate and calcium acetate implants) showed more effects on the bone tissue responses than RBM surface in initial period of implantation. In addition, CMP showed a tendency to promote bone tissue responses.
Objectives: The objective of this study was to investigate the antioxidant effects of manufactured Wen-pi-tang-Hab-Wu-ling-san (WHW$^{(R)}$) in vitro. Methods: WHW$^{(R)}$ was prepared by the pilot manufacture of WHW water extract from a GMP system appointed company. Antioxidative activities were determined by in vitro tests as follows: the scavenging activities of oxygen free radicals including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, superoxide anion, hydrogen peroxide and nitric oxide radicals, as well as ferrous ion chelating capacity and Trolox equivalent antioxidant capacity (TEAC). Results: WHW$^{(R)}$ significantly scavenged oxygen free radicals such as DPPH (IC$_{50}$=115.28 $\pm$ 0.25 $\mu$g/$m\ell$), superoxide anion (IC$_{50}$=8.56 $\pm$ 0.08 $\mu$g/$m\ell$), hydrogen peroxide (IC$_{50}$=240.36 $\pm$ 3.41 $\mu$g/$m\ell$) and nitric oxide (IC$_{50}$=162.28 $\pm$ 0.21 $\mu$g/$m\ell$) radicals. WHW$^{(R)}$ also showed ferrous ion chelating activity (IC$_{50}$=543.19 $\pm$ 4.85 $\mu$g/$m\ell$) and Trolox equivalent effects (IC$_{50}$=45.311 $\mu$g/$m\ell$) in TEAC and ORAC assay, respectively. Conclusion: This study demonstrates that WHW$^{(R)}$ has strong antioxidative properties through free radical scavenging activity. These data suggest that WHW$^{(R)}$ be used as an antioxidant agent.
In the present study, we characterized the angiotensin II (AII)-induced relaxations in the phenylephrine-precontracted rabbit mesenteric arteries with endothelium. 1) AII-induced relaxation was consistently observed in the rabbit mesenteric arteries with and without endothelium, but not in the aortic segment with endothelium. 2) AII-induced endothelium-dependent relaxation was markedly inhibited by $N^w-nitro-L-arginine$ (L-NNA, $100\;{\mu}M$), methylene blue ($10\;{\mu}M$) and LY83583 ($10\;{\mu}M$), respectively. 3) Inhibition of cyclooxygenase with indomethacin ($10\;{\mu}M$) strongly decreased the vasorelaxant response to AII irrespective of the presence of endothelium. 4) 7-Ethoxyresorufin ($1\;{\mu}M$) and clotrimazole ($1\;{\mu}M$), inhibitors of cytochrome P-450-dependent arachidonic acid metabolism, greatly attenuated the vasodilator response to AII. 5) Carbacyclin, arachidonic acid and prostaglandin $F_{2{\alpha}}$ ($PGF_{2{\alpha}}$) caused concentration-dependent relaxations in the mesenteric artery with endothelium, which were inhibited by L-NNA and methylene blue. 6) AII and $PGF_{2{\alpha}}$ significantly stimulated cyclic GMP formation in the mesenteric arteries with endothelium, which was inhibited by L-NNA and methylene blue, respectively. 7) AII enhanced synthesis of $PGF_{2{\alpha}}$ and 6-keto $PGF_{1{\alpha}}$ from the arterial segments with endothelium, which was inhibitable by indomethacin, but not by L-NNA. In conclusion, the vasorelaxant responses to AII of the rabbit mesenteric artery with endothelium are subserved by arachidonic acid and its metabolites produced via activation of cyclooxygenase and cytochrome P-450 enzyme as well as by nitric oxide.
Yuldahansotang(YH) has been used in Sasang(四象) constitution medicine for many years as a therapeutic agent for cerebral disease. The effect of YH on the vascular system is not known. The purpose of this study was to determine the effect of YH on blood pressure, regional cerebral blood flow(rCBF) and pial arterial diameter of rats. 1. Blood pressure decreased by YH in rats. 2. rCBF was increased by YH in a dose-dependent manner. 3. Pretreatment with propranolol, methylene blue and indomethacin significantly inhibited YH induced increase in rCBF. 5. Blood pressure increased by Radix Puerariae(RP) and Radix Ligustici Tenuissimae(RLT) but Radix Scutellariae(RC) decreased blood pressure in rats. 6. rCBF was increased by RP and RLT in a dose-dependent manner but RC decreased low dosage, and RC increased high dosage. 7. Pial arterial diameter was increased by YH in a dose-dependent manner. 8. Pretreatment with propranolol significantly inhibited the increased in pial arterial diameter induced by YH. These results suggest that YH causes a diverse response of blood pressure, regional cerebral blood flow(rCBF) and pial arterial diameter. The increase in rCBF is also mediated by prostaglandins, cyclic GMP and adrenergic ${\beta}$ receptor and the increase in pial arteral diameter is mediated by adrenergic ${\beta}$ receptor.
The therapeutic efficacy of xylamine in the field of psychological medicine has been recognized for years and the drug is used to treat depression and some other conditions, but little is known about its mechanism of action on vascular system. Therefore, the present study was designed to investigate the influence of xylamine on the contractile responses of isolated rat thoracic arteries to phenylephrine(PE) and potassium chloride(KCl). Xylamine produced a concentration-dependent relaxation in PE-precontracted endothelium intact(+E) rat aortic rings, but not in a KCl-precontracted aortic rings. Also, xylamine inhibited the PE-induced contraction in concentration-dependent manner, but not in the high KCl-induced contraction in +E rings. This concentration-dependent inhibition was suppressed by the removal of the endothelium (-E). The inhibitory effects of xylamine($0.3{\mu}M$) on the PE-induced contractions were suppressed by N(G)-nitro-L-arginine(L-NNA), N(omega)-nitro-L-arginine methyl ester(L-NAME), aminoguanidine, dexamethasone, methylene blue, 1H-[1,2,4]oxadiazolo [4,3-a]quinoxalin-1-one(ODQ), indomethacin, ryanodine, tetrabutylammonium(TBA), lidocaine, procaine and 0 mM extracellular $Na^+$, but not by 2-nitro-4-carboxyphenyl-n,n-diphenylcarbamate(NCDC), lithium, nifedipine, verapamil, 0 mM extracellular $Ca^{2+}$, glibenclamide and clotrimazole. These findings suggest that xylamine could act as a vasorelaxant and direct inhibitor of arterial contraction. This vasorelaxation involves an endothelial nitric oxide (NO)/cGMP (guanosine 3',5'-cyclic monophosphate) pathway or cyclooxygenase system, and an interference with $Ca^{2+}$ release, TBA-sensitive $Ca^{2+}$-activated $K^+$ channels and $Na^+$$ channels.
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