Changes of silk protein compositions of average molecular weight (Mw) and free amino acid composition to different solubility conditions were studied by SDS-polyacrylamide electropholesis, gel permeation chromatography (GPC), and free amino acid analysis method. We can not detected average molecular weight distribution of different hydrochloric acid (HCl) conditions as SDS-polyacrylamide method, but as using GPC method, molecular weight distribution of 2N-HCl, 1N-HCl and 0.5N-HCl (3 hrs at $110^{\circ}C$ treated) are confirmed Mw 800, 1,500 and 3,700, respectively. The average molecular weight of calcium chroride and calcium chloride-enzyme treated samples are shown Mw 46,800 and 12,500, respectively. The degree of hydrolysis and the composition of the free amino acid in the fibroin hydrolysates effected significantly composition of free amino acids of the fibroin powder. The increase of the degree of hydrolysis and ratio of free amino acids and oligopeptides were found to be directly related to the concentration of hydrochloric acid and treatment of enzyme, resulting in the increase of water solubility.
The addition of one mole of zinc chloride to 2.33 moles of sodium borohydride in tetrahydrofuran at room temperature gave a clear chloride-free supernatant solution of zinc borohydride after stirring three days and standing at room temperature.The approximate rates and stoichiometry of the reaction of zinc borohydride with 54 selected organic compounds were determined in order to test the utility of the reagent as a selective reducing agent. Aldehydes and ketones were reduced rapidly, aromatic ketones being somewhat slowly, and the double bond of cinnamaldehyde was not attacked. Acyl halides were reduced rapidly within one hour, but acid anhydrides were reduced at a moderate rate. Carboxylic acids, both aliphatic and aromatic, were slowly reduced to alcoholic stage. Esters were inert to this reagent but a cyclic ester, γ-butyrolactone, was slowly attacked. Primary amides were reduced slowly with partial evolution of hydrogen, whereas tertiary amides underwent neither reduction nor hydrogen evolution. Epoxides and nitriles were all inert, as well as nitro, azo, and azoxy compounds. Cyclohexanone oxime and phenyl isocyanate were reduced slowly but pyridine was inert. Disulfide, sulfoxide, sulfone and sulfonic acids were stable to this reagent.
Renal arterial infusion of renotropic agents has been a very useful technique in the renal function studies. This type of experiments have usually been conducted in the large animals such as dogs and sheep. In these animals a catheter can be placed in the site without much disturbances of renal blood flow. Rabbits as an experimental model, however, caused a disturbances of renal blood flow by a catheterization of renal artery by its properties. Therefore we have developed a new technique that allows a simple and selective access to one side of renal arteries and the other as a control, without any disturbances of renal function. The distance between the both bifurcations of renal arteries on abdominal aorta is about 7 mm. To locate the tip of catheter on one side renal artery, ascending cannulation performed via femoral artery was done. We did an experiment with the technique to clarify the effect of calmodulin inhibitor on the renal function. One of the phenothiazine derivatives, trifluoperazine known as a powerful calmodulin inhibitor. Trifluoperazine, actual dose ranges of $2.76-5.20\;ug\;{\cdot}\;kg^{-1}\;{\cdot}\;min^{-1}$, increased urine volume and glomerular filtration rate significantly. Significant increases in urinary excretion of sodium, chloride and potassium were found. Fractional excretion of sodium and free water clearance increased significantly. These data suggest that this new technique is very useful in field of renal physiology and that striking effect of trifluoperazine on the renal function may be caused by increasing the renal hemodynamics, and by the inhibition of sodium, chloride and potassium reabsorption in the renal tubules.
The contact angle on powder hydrophobic polymer PMMA(polymethylmethacrylate), PVC(poly vinyl chloride) and hydrophilic metal aluminium was measured by wicking method on the base of Washburn equation for water, glycerol, formamide, diiodomethane, 1,1,2,2-tetrabromoethane, 1-bromonaphthalene, acetone, chloroform, benzene, ethanol, methanol, and for anionic surfactant SDS(sodium dodecyl sulfate) and cationic surfactant CATB(cetyl trimethylamonium bromide) aqueous solution with concentrations. The values of ${\gamma}_c$ estimated by Zisman plot and adhesion tension plot for contact angle with organic liquids, SDS, and CTAB aqueous solutions were 43.5, 28.3, and $36.2mNm^{-1}$ for PMMA and 44.2, 25.0, and $34.8mNm^{-1}$ for PVC, respectively. From the results, it was considered that wettability and characteristics of solid surface were transformed by adding surfactant. The surface free energy, ${\gamma}_s$ and its components of PMMA, PVC, and aluminium were determined using harmonic equation.
As a part of our search for novel antioxidants from the seaweeds, we have investigated radical scavenging effect for their crude extracts using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, authentic peroxynitrite, and 3-morpholinsydnonimine (SIN-1), a peroxynitrite-generating species in vitro. Thirty-four seaweeds were screened for $ONOO^-$ and DPPH radical scavenging activities. A potent inhibitory effect against peroxynitrite generated by SIN-1 at $5{\mu}g/ml$ of methanol extracts was observed in order of Ishige okamurae(95.3%), Sargassum hemiphyllum(90.2%), Symphyocladia latiuscula(89.6%), Porphyra suborbiculata(86.7%), and Gelidium amamsii(85.9%), Also, a significant scavenging effect against direct authentic peroxynitrite was revekaled for methanol extracts of Ishige okamurae(66.2%) and Sargassum hemiphyllum(55.2%) and the acetone/methylene chloride(1:1) extract of Gigatina tenella (61.0%). In our measurement for evaluating the capacity to scavenge the stable free radical of DPPH, acetone/methylene chloride(1:1) extracts of Symphyocladia latiuscula, Gloiopeltis furcata, and Sargassum thunbergii and the methanol extract of Sargassum sp. showed an inhibitory potency of 85.8%, 82.8%, 74.1%, and 64.0%, respectively.
Journal of Physiology & Pathology in Korean Medicine
/
v.30
no.2
/
pp.101-108
/
2016
This study is conducted to investigate vasorelaxant effect of Corni Fructus(CF) on rabbit carotid artery. To determine vasorelaxant effect of CF on rabbit carotid artery, arterial sections with intact or removed endothelium were used in this organ bath study. After being contracted by phenylephrine(PE), arterial sections were treated with CF extract in a dose-dependent manner. To identity its mechanism, the contracted arterial sections by PE were pretreated with indomethacin(IM), tetraethylammonium chloride(TEA), Nω-nitro-L-arginine(L-NNA) or methylene blue(MB) and 1.0 ㎎/㎖ CF extract. We also studied to confirm the effect on influx of extracellular calcium chloride(Ca2+) of the CF extract in rabbit carotid artery. To measure the cytotoxicity of the CF extract, cell viability of human umbilical vein endothelial cell(HUVEC) was measured by MTT assay. Generation of nitric oxide(NO) was also measured by Griess reagent. The arterial sections with intact endothelium were relaxed significantly by CF extract, but this effect was inhibited in the arterial sections with damaged endothelium. The vasorelaxant effect was inhibited significantly when arterial sections were pretreated with IM, TEA, L-NNA, MB. In Ca2+-free krebs solution, increasing of arterial contraction by Ca2+ was also inhibited by CF significantly. The treatment of CF extract increased NO concentration in HUVEC. This study suggested that the vasorelaxant effect of CF extract would be related with endothelium derived relaxing factor(EDRF) such as NO, prostacyclin(PGI2), endothelium derived hyperpolarization factor(EDHF).
Journal of Physiology & Pathology in Korean Medicine
/
v.19
no.6
/
pp.1622-1628
/
2005
This study was undertaken to define the effect of SoPung-Tang extract on hypertension in spontaneous hypertensive rat and norepinephrine- induced arterial contraction in rabbit. In order to investigate the effect of SoPung-Tang extract on contracted rabbit carotid arterial strips, transverse strips with intact or damaged endothelium were used for the experiment using organ bath. To analyze the mechanism of SoPung-Tang extract-induced relaxation, SoPung-Tang extract infused into contracted arterial strips induced by norepinephrine after treatment of indomethacin, Nu-nitro-L-arginine, methylene blue or tetraethylammonium chloride. Blood pressure was significantly decreased five days after administration of SoPung-Tang extract. SoPung-Tang extract relax arterial strip with endothelium contracted by norepinephrine, but in the strips without endothelium, SoPung-Tang extract- induced relaxation was significantly inhibited. SoPung-Tang relax arterial strip contracted by norepinephrine, but in the strips contracted by high $K^+$, SoPung-Tang extract-induced relaxation was significantly inhibited. The endothelium-dependent relaxation induced by SoPung-Tang extract was decreased by the pre-treatment of $N{\omega}$-nitro-L-arginine or methylene blue, but it was not observed in the strips pre-treated with indomethacin or tetraethylammonium chloride. When $Ca^{2+}$ was applied, the strips which were contracted by norepinephrine in a $Ca^{2+}$-free solution, arterial contraction was increased. But pre-treatment of SoPung-Tang extract inhibited contractile response to $Ca^{2+}$. We suggest that SoPung-Tang could be applied effectively for hypertension and may suppress influx of extra-cellular $Ca^{2+}$ through the formation of nitric oxide in the vascular endothelial cells.
Objectives : This experiments were performed to determine the effect of Linderae Radix extract on norepinephrine-induced arterial contraction in rabbit. Methods : In order to investigate the effect of Linderae Radix extract on contracted rabbit carotid arterial strips, transverse strips with intact or damaged endothelium were used for the experiment using organ bath. To analyze the mechanism of Linderae Radix extract-induced relaxation, Linderae Radix extract infused into contracted arterial strips induced by norepinephrine after treatment of indomethacin, tetraethylammonium chloride, $N{\omega}-nitro-L-arginine$ or methylene blue. Results : Linderae Radix extract relax arterial strip with endothelium contracted by norepinephrine, but in the strips without endothelium, Linderae Radix extract-induced relaxation was significantly inhibited. Linderae Radix extract-induced relaxation was decreased by the pre-treatment of $N{\omega}-nitro-L-arginine$ or methylene blue, but it was not observed in the strips pre-treated with indomethacin or tetraethylammonium chloride. When $Ca^{2+}$ was applied, the strips which were contracted by norepinephrine in a $Ca^{2+}$-free solution, arterial contraction was increased. But pre-treatment of Linderae Radix extract inhibited contractile response to norepinephrine and $Ca^{2+}$. Conclusions : We suggest that Linderae Radix may suppress influx of extra-cellular $Ca^{2+}$ through the formation of nitric oxide, and release of intra-cellular $Ca^{2+}$.
Objectives : This experiments were performed to determine the effect of OYakSoonGi-San extract on hypertension in spontaneous hypertensive rat and norepinephrine-induced arterial contraction in rabbit. Methods : In order to define the effect of OYakSoonGi-San extract on contracted rabbit carotid arterial strips, transverse strips with intact or damaged endothelium were used for the experiment using organ bath. To analyze the mechanism of OYakSoonGi-San extract-induced relaxation, OYakSoonGi-San extract infused into contracted arterial strips induced by norepinephrine after treatment of indomethacin, $N{\omega}-nitro-L-arginine$, methylene blue or tetraethylammonium chloride. Results : Blood pressure was significantly decreased five days after administration of OYakSoonGi-San extract. The relaxation effect of OYakSoonGi-San extract was dependent on the presence of endothelium, showing that OYakSoonGi-San extract-induced relaxation was not observed in the strips without endothelium. Also OYakSoonGi-San extract-induced relaxation was significantly inhibited in arterial strips which were contracted by high $K^+$. OYakSoonGi-San extract-indeced relaxation was significantly inhibited by the pre-treatment of $N{\omega}-nitro-L-arginine$ or methylene blue, but it was not observed in the strips pre-treated with indomethacin or tetraethylammonium chloride. When additive application of $Ca^{2+}$ in arterial strips which were pre-contracted by norepinephrine in a $Ca^{2+}$-free solution, arterial contraction was increased. But contractile response to $Ca^{2+}$ was attenuated by pre-treatment of OYakSoonGi-San extract. Conclusions : These results demonstrated that OYakSoonGi-San could be applied effectively to hypertension and may inhibit agonist-induced contraction through an decrease influx of extra-cellular $Ca^{2+}$ by the formation of nitric oxide in the vascular endothelial cells.
Objectives : The purpose of this study was to investigate the endothelium-dependent vasorelaxation effect of Cynomorii Herba(CH) extract on contracted rabbit carotid artery.Methods : To clarify the vasorelaxation effect of CH extract, arterial strips with intact was used, to endothelium -dependent vasorelaxation effect of CH extract, arterial strips damaged endothelium was used for experiment using organ bath. Arterial strips was contracted with phenylephrine(PE) before treated with CH extract(0.01, 0.03 and 0.1 ㎎/㎖). To study mechanisms of CH-induced vasorelaxation effect, CH extract infused into arterial rings after treatment by indomethacin(IM), tetraethylammonium chloride(TEA), Nω-nitro-L-arginine (L-NNA), methylene blue(MB) for comparing with non-treated. And calcium chloride(Ca2+) 1 mM was treated into precontracted arterial ring induced by PE after treatment of CH extract in Ca2+-free krebs solution. Cytotoxic activity of CH extract on human umbilical vein endothelial cell(HUVEC) was measured by MTT assay, and nitric oxide(NO) concentration was measured by Griess reagent.Results : PE-induced arterial strips was significantly relaxed, but the damaged endothelium arterial ring wasn't relaxed by CH extract. Pretreatment of IM, TEA didn't inhibit the vasorelaxation of CH extract, but pretreatment of L-NNA, MB inhibited the vasorelaxation of CH extract. Pretreatment of CH extract reduced the increase of contraction by influx of extracellular Ca2+ in contracted arterial ring induced by PE, CH extract increased nitric oxide concentration on HUVEC significantly.Conclusions : This study shows that CH extract have the vasorelaxation effect by blocking the influx of extracellular Ca2+ through the activating NO-cGMP system.
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