To investigate the endothelium dependent vascular reactivity of the systemic arterial and the pulmonary arterial system in acute renal hypertensive rats of 2-kidney, 1-ligation type (RHRs), acetylcholine (ACh)-induced vasodilation and depressor effects were evaluated in isolated arteries and in vivo, respectively, in the presence and absence of functional endothelium. ACh $(10^{-5}\;M)$ relaxed the intact thoracic aortas from RHRs and normotensive rats (NRs), but the effect was significantly smaller for those from RHRs (34 and 86%, respectively, p<0.01). ACh-induced vasodilation was completely abolished after removal of endothelial cell or pretreatment with EDRF inhibitors, L-NAME and MB, indicative of its dependence on intact endothelial or EDRF function. ACh also induced vasorelaxation of the intact pulmonary arteries from RHRs and NRs; however, unlike the effects on the thorcic aorta, no significant difference in amplitude was noted between two groups. ACh $(0.1{\sim}10\;{\mu}g/kg,\;i.v.)$ reduced mean systemic arterial pressure in anesthetized RHRs and in NRs to the similar magnitude (% change: 39 and 46% at $10\;{\mu}g/kg$, respectively) and these hypotensive effects were significantly decreased after pretreatment with L-NAME (30 mg/kg, i.v.). Deprssor effects of ACh on mean pulmonary arterial pressure were similar in RHRs and NRs with and without pretreatment of L-NAME. However, in both NRs and RHRs, the depressor effects of ACh on mean pulmonary arterial pressure were significantly reduced compared with those for mean systemic arterial pressure, and the increment of mean pulmonary arterial pressure noted after L-NAME $(0.1{\mu}100\;mg/kg,\;i.v.)$ was significantly smaller than that for mean systemic arterial pressure. These results indicate that in RHRs the endothelial cell function was impaired, at least in part, in systemic arterial system, but not in pulmonary arterial system, and both ACh-evoked and basal release of EDRF was less in the pulmonary arterial system than in systemic arterial system of both NRs and RHRs.
Kwon, Jin Gwan;Kim, Jin Kyu;Seo, Changon;Hong, Seong Su;Ahn, Eun-Kyung;Seo, Dong-Wan;Oh, Joa Sub
Journal of the Korean Society of Food Science and Nutrition
/
v.45
no.1
/
pp.61-67
/
2016
An HPLC analysis method was developed for standard determinations of chlorogenic acid, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 4,5-di-O-caffeoylquinic acid as functional health materials in Ligularia fischeri extract. HPLC was performed on a $C_{18}$ Kromasil column ($4.6{\times}250mm$, $5{\mu}m$ column) with a gradient elution of 0.1% (v/v) trifluoroacetic acid and acetonitrile at a flow rate of 1.0 mL/min at $30^{\circ}C$. The analytes were detected at 330 nm. The HPLC method was validated in accordance with the International Conference on Harmonization guideline of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limits of detection and quantitation for the four compounds were 3.0~14.6 and $9.2{\sim}44.4{\mu}g/mL$, respectively. Calibration curves showed good linearity ($r^2$ > 0.999), and the precision of analysis was satisfied (less than 0.9%). Recoveries of quantified compounds ranged from 98.96 to 101.81%. This result indicates that the established HPLC method is very useful for the determination of marker compounds in Ligularia fischeri leaf extracts.
It is essential to evaluate the quality of Hanji record paper without damaging the record paper by previous destructive methods. The samples were Hanji record paper produced in the 1900s. Near-infrared (NIR) spectrometer was used as a non destructive method for evaluating the quality of record papers. Fourier transform (FT) spectrometer was used with 12,500 to 4,000 $cm^{-1}$ wavenumber range for quantitative analysis and it has high accuracy and good signal-to-noise ratio. The acidity and moisture content of Hanji record paper were measured by integrating sphere as diffuse reflectance type. The acidity (pH) of chemical factors as a quality evaluated factor of Hanji was correlated to NIR spectrum. The NIR spectrum was pretreated to obtain the coefficients of optimum correlation. Multiplicative scatter correction (MSC) and First derivative of Savitzky-Golay were used as pretreated methods. The coefficients of optimum correlation were calculated by PLSR (partial least square regression). The correlation coefficients ($R^2$) of acidity had 0.92 on NIR spectra without pretreatment. Also the standard error of prediction (SEP) of pH was 0.24. And then the NIR spectra with pretreatment would have better correlation coefficient ($R^2$ = 0.98) and 0.19 as SEP on pH. For moisture contents, the linearity correlation without pretreatment was higher than the case with pretreatment (MSC, $1^{st}$ derivative). As the best result, the $R^2$ was 0.99 and SEP was 0.45. This indicates that it is highly proper to evaluate the quality of Hanji record papers speedily with integrated sphere and FT NIR analyzer as a non-destructive method.
The aerial parts of Sajabalssuk (Artemisia princeps PAMPANINI, Sajabalssuk) was extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH and $H_2O$, successively. From the EtOAc fraction, three cycloartane-type triterpnoids and one ursane-type triterpenoid were isolated through the repeated silica gel, ODS and Sephadex LH-20 column chromatographies. From the results of physico-chemical data including NMR, MS and IR, the chemical structures of the triterpenoids were determined as wrightial (1), wrightial acetate (2), 27-norcycloart-20(21)-ene-25-al-3${\beta}$-ol acetate (3) and ursolic acid (4). No report has been found for isolation of compound 3 in the literature so far, and compounds 1, 2 and 3 were the first to be isolated from Sajabalssuk (Artemisia princeps PAMPANINI, Sajabalssuk). Also, compound 1 showed Acyl-CoA:Cholesterol acyltransferase (hACAT-1) and hACAT-2 inhibitory activity with the $IC_{50}$ values of 33.0 and 45.0 ${\mu}g/ml$, respectively. Compounds 2 and 3 inhibited hACAT-1 activity with the $IC_{50}$ values of 12.0 and 16.0 ${\mu}g/ml$, respectively.
Liver microsomal epoxide hydrolase (mEH) is active in the detoxification of epoxide-containing reactive intermediate. Previous studies in this laboratory have shown that thiazole and pyrazine are efficacious inducers of mEH in rats with large increases in mEH mRNA levels (Carcinogensis, Kim et al, 1993). mEH was purified to electrophoretic homogeneity from thiazole-induced rat hepatic microsomes using DEAE-cellulose column chromatography whereas another protein $({\sim}43\;kDa)$ was co-purified with mEH from pyrazine-induced rat hepatic micrsomes (200 mg/kg body weight/day, ip, 3d). The antibody raised from a rabbit against mEH protein purified from thiazole-induced rat hepatic microsomes appeared to specifically recognize mEH protein in rat hepatic microsomes, as assessed by immunoblotting analysis. Immunoblotting analyses revealed a 10- and 7-fold increase in mEH levels in the hepatic microsomes isolated from thiazole- and pyrazine-treated rats, respectively. Moreover, immunoblotting analysis showed cross-reactivity of the mEH antibody with a 43 kDa protein in pyrazine-induced rat hepatic microsomes and with co-purified 43 kDa protein in purified fractions. The ratio between the 43 kDa protein and mEH in pyrazine-induced rat microsomes or in purified fractions was ${\sim}1$ to 15. N-terminal amino acid sequence analysis of both purified rat mEH and 43 kDa protein revealed that 10 out of 12 amino acids in N-terminus of the 43 kDa protein were identical with the mEH sequence with two amino acid residues of the 43 kDa protein undetermined. Either thiazole or pyrazine treatment, however, failed to increase the levels of mEH protein in rabbits while pyrazine caused elevation of the 43 kDa protein in this species, as determined by irnrnunoblotting analysis. These results demonstrated that treatment of rats with either thiazole or pyrazine causes elevation in hepatic mEH expiession whereas pyrazine treatment results in induction of another mEH-related 43 kDa protein and that a distinct species difference exists between rats and rabbits in the induction of mEH by these xenobiotics.
In de novo biosynthesis of the extracellulor enzymes-proteinsaes, alpha and gluc-amylases during the synchronized differentiation of Aspergillus niger in submerged culture and surface liquid culture were investigated. Gluc-amylase was synthesized in the stage of presporulation in which phialide formation is involved. Proteinase was synthesized both in the stages of conidiophore formation and presporulation. Alpha-amylase was synthesized during presporulation and sporulation stages, the activity of enzyme lasted for seven days on surface liquid culture. It seemed that the synthesis was occured in de novo partly repressed by the catabolite, and its nature was found to be constitutive since it is produced in non-starch medium. Polyacrylamide gel electrophoresis have shown that presporulating and sporulating body produced diverse types of the proteins whereas the earlier stages of vegetative body showed simpler profiles. The uptake of C-14 uracil into RNA and C-14 glutamate into protein were shown to be vigorous in presporulating body rather than those in sporulating body. Coincidence of alpha-amylase biosynthesis in de novo and sporulation may be significant in the study of differentiation in which gene expression is involved.
The influence of incubation temperature, pH and media components on bacteriocin production by Lactobacillus plantarum K11 were investigated. The highest activity was observed in MRS broth, but no bacteriocin activity was obtained in TSB. The bacteriocin was produced from the exponential growth phase and its activity also reached a maximum in MRS broth, but then dropped after 16 hr because of degradation by extracellular proteolytic enzymes or exhaustion of medium nutrients. The optimal temperature and pH for production of bacteriocin were $37^{\circ}C$ and pH 7.0 in MRS broth, respectively. The addition of 0.5 or 1.0% glucose and $0.5{\sim}1.5%$ lactose to MRS resulted in the increase of the bacteriocin production. With 0.5% NaCl and $K_2HPO_4$, the activities were significantly higher than that of control, respectively. However, increasing nitrogen sources such as beef extract, casein, and tryptone and salts such as $NH_4PO_4$, $MgSO_47H_2O$, and $MnSO_4H_2O$, had detected a negative influence upon the bacteriocin production. Consequently, because the bacteriocin produced by L. plantarum K11 was affected by various incubation conditions, the bacteriocin activity of L. plantarum K11 applied in food as a novel starter will be dependent on environmental factors such as fermentation conditions and food ingredients.
Hyun, Sun Hee;Kim, Eun Sun;Lee, Sang Min;Kyung, Jong Soo;Lee, Sang Myung;Lee, Jong Won;Kim, Mee Ree;Hong, Jin Tae;Kim, Young Sook
Journal of the Korean Society of Food Science and Nutrition
/
v.43
no.11
/
pp.1665-1673
/
2014
The objective of this study was to compare the compositions and immuno-enhancing effects of 6-year-old red ginseng powder (RGP) with those of its fractions. RGP was subjected to extraction with 100% ethanol to obtain an ethanol fraction (E) and residue 1 (R1). Then, R1 was subjected to extraction with distilled water to obtain water fraction (W) and residue 2 (R2). Chemical compositions were as follows: 4.94% acidic polysaccharides and 1.56% ginsenosides (amounts of Rg1, Re, Rf, Rg2, Rb1, Rc, Rd, and Rg3) in RGP, 0.11% acidic polysaccharides and 6.99% ginsenosides in E, 4.93% acidic polysaccharides and 0.40% ginsenosides in R1, 0.50% acidic polysaccharides and 0.30% ginsenosides in R2, and 7.46% acidic polysaccharides and 0.61% ginsenosides in W. Immuno-enhancing effects of fractions from RGP were examined based on suppression of immune responses by cyclophosphamide. In the first fraction test, the antibody response to SRBCs increased significantly in the R1-treated group, but not the E-treated group. In the second fraction test, W showed higher immuno-enhancing effect than R1 and R2. W, which contained the highest amount of acidic polysaccharides, restored numbers of T and B cells, macrophages, as well as $CD4^+$ and $CD8^+$ T cells in the spleen suppressed by cyclophosphamide. These results suggest that acidic polysaccharides from red ginseng may be more effective than saponin in enhancing immune functions and reducing immunotoxicity of cyclophosphamide.
The subventricular zone (SVZ) in the brain contains neural stem cells (NSCs) that generate new neurons throughout one's lifetime. Many extracellular and intracellular factors that affect cell proliferation and neuronal differentiation of NSCs are already well-known. Recently, L-type calcium channels have been reported to regulate neural development and are present in NSCs, differentiating neuroblasts, and mature neurons in the SVZ. Nifedipine, a blocker of L-type calcium channels, has been long used as a therapeutic drug for hypertension. However, studies on the use of nifedipine to inhibit L-type calcium channels of NSCs are lacking. Herein, we treated NSCs cultured from mouse postnatal SVZ with nifedipine during neuronal differentiation. Nifedipine increased the number of Tuj1-positive neurons but did not significantly change the number of Olig2-positive oligodendrocytes. Nifedipine increased cell division during early differentiation, which was detected using the 5-ethynyl-2'-deoxyuridine incorporation assay and immunocytochemistry assessment by staining the cells with phosphorylated histone H3, a mitosis marker. Nifedipine increased the transcription of Dlx2, a neurogenic transcription factor, and the level of Mash1, a marker for early neurogenesis. In addition to nifedipine, verapamil, which is also an L-type calcium channel blocker, showed a slight increase in neurogenesis, but its statistical significance was very low. In contrast, pimozide, a T-type calcium channel blocker, did not affect neurogenesis, although T-type calcium channel genes Cav3.1, Cav3.2, and Cav3.3 were expressed. In summary, nifedipine might promote the neuronal fate of NSCs during early differentiation and calcium signaling through L-type calcium channels might be involved in neuronal differentiation, especially during the early stages of differentiation.
In the present study, we investigated the biological activities of Xylosma congesta leaf ethanol extract (XCO) using a variety of in vitro and cell culture model systems for anti-melanogenic, anti-wrinkle, anti-inflammatory and anti-oxidant activities. First, XCO markedly inhibited ${\alpha}$-melanocyte stimulating hormone-stimulated melanin synthesis in B16F10 cells. Secondly, XCO marginally induced procollagen synthesis in CCD-986SK cells. Thirdly, XCO dose-dependently suppressed lipopolysaccharide-induced nitric oxide (NO) production in RAW 264.7 cells. XCO did not affect cell viability at different concentrations used in this study, indicating that XCO-mediated inhibition of melanin, procollagen and NO synthesis is not mediated by cytotoxicity. Finally, XCO was found to exert anti-oxidant effect. Taken together, these findings demonstrate for the first time that XCO possesses anti-melanogenic, anti-wrinkle, anti-inflammatory and anti-oxidant activities, and suggest further evaluation and development of XCO as a functional supplement or cosmetic that may be useful for whitening skin, reducing wrinkles and treating inflammatory responses.
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