• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.5
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• pp.1315-1321
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• 2008

Chan-Su (Venenum bufonis) has long been for a variety of other purposes including treatment of inflammation in the folk medicine recipe. Since nitric oxide (NO) is one of the major inflammatory parameters, we first studied the effects of Chan-Su on NO production in lipopolysaccharide (LPS)-stimulated BV2 microglial cells, Chan-Su inhibited the secretion of NO in BV2 microglial cells, without affecting cell viability, The protein level of inducible nitric oxide synthase (iNOS) was decreased by Chan-Su, And Chan-Su also inhibited production of prostaglandin E2 (PGE2) and expression of cyclooxygenase (COX)-2. Proinflammatory cytokines, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$ and IL-12, were inhibited by Chan-Su in a dose-dependent manner. And Chan-Su inhibited the degradation of ${IkB-\alpha}$, which was considered to be inhibitor of nuclear factor $(NF)-{\kappa}B$, one of a potential transcription factor for the expression of iNOS, COX-2 and proinflammatory cytokines. These results suggest that Chan-Su could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of $I{\kappa}B-{\alpha}$ degradation.

• Journal of Life Science
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• v.24 no.12
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• pp.1269-1275
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• 2014

TALEN is a newly developed gene engineering method to knock out specific genes. It contains a DNA binding domain and a Fok1 nuclease domain in the TALEN plasmid. Therefore, the engineered TALEN construct can bind to any region of genomic DNA and cut the target nucleotide, thereby inducing mutation. In this study, we constructed two TALEN constructs targeted to a protein initiation codon (DBEX2) or the 25th upstream region (DBPR25) to enable mRNA synthesis of SETDB1 HMTase. We performed the TALEN cloning in two steps. The first step was from module vectors to pFUS array vectors. We confirmed successful cloning with a colony PCR experiment and Esp31 restriction enzyme digestion, which resulted in a smear band and a 1 Kb insert band, respectively The second step of the cloning was from a pFUS array vector to a mammalian TALEN expression vector. The engineered TALEN construct was sequenced with specific primers in an expression vector. As expected, a specific array from the module vectors was shown in the sequencing analysis. The specific module sequences were regularly arrayed in every 100 bp, and SETDB1 expression totally disappeared in the TALEN-DBEX2 transfection. PCR amplification targeting of DBEX2 was performed, and the PCR product was digested with a T7E1 restriction enzyme. The expression of SETDB1 was down-regulated in the TALEN-DBPR25 transfection. Morphological changes were also observed in the two TALEN constructs with transfected HeLa cells. These results suggest that the engineered TALEN constructs in two strategic approaches are very useful to knock-out of the SETDB1 gene and to study gene function.

• Journal of Life Science
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• v.23 no.8
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• pp.1064-1072
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• 2013

Circadian rhythm is controlled by hormonal oscillations governing the physiology of all living organisms. In mammals, the main function of the pineal gland is to transform the circadian rhythm generated in the hypothalamic suprachiasmatic nucleus into rhythmic signals of circulating melatonin characterized by a largely nocturnal increase that closely reflects the duration of night time. The pineal gland has lost direct photosensitivity, but responds to light via multi-synaptic pathways that include a subset of retinal ganglion cells. Rhythmic control is achieved through a tight coupling between environmental lighting and arylalkylamine-N-acetyltransferase (AANAT) expression, which is the rhythm-controlling enzyme in melatonin synthesis. Previous studies on the nocturnal expression of AANAT protein have described transcriptional, post-transcriptional, and post-translational regulatory mechanisms. Molecular mechanisms for dependent AANAT expression provide novel aspects for melatonin's circadian rhythmicity. Extensive animal research has linked pineal melatonin for the expression of seasonal rhythmicity in many mammalian species to the modulation of circadian rhythms and to sleep regulation. It has value in treating various circadian rhythm disorders, such as jet lag or shift-work sleep disorders. Melatonin, also, in a broad range of effects with a significant regulation influences many of the body's physiological functions. In addition, this hormone is known to influence reproductive, cardiovascular, and immunological regulation as well as psychiatric disorders.

• Journal of Life Science
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• v.29 no.12
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• pp.1321-1328
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• 2019

Intermediate-wavelength solar radiation, also known as ultraviolet B (UVB: 290-320 nm) radiation, may cause premature aging and oxidative damage-dependent skin cancer in humans. UVB-induced formation of reactive oxygen species (ROS)-often a consequence of excessive exposure to these rays-could activate matrix metalloproteinases (MMPs) such as MMP-1 and MMP-3. These enzymes break down type I collagen in human fibroblasts. In this study, we assessed the antioxidant and anti-aging effects of ethyl acetate extract of blueberry (EEB). An antioxidant test in blueberries evaluated ROS production using CCD-986sk cells and DPPH assay. In order to evaluate the anti-wrinkle efficacy of blueberries, the MMP-1 production and type 1 procollagen synthesis evaluated and the expression of MMP 1, 3 were tested through Western blot and RT- PCR. EEB exhibited 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and reduced the production of UVB-induced ROS. Also, EEB inhibited UVB-induced processes associated with photoaging and skin cancer, such as reduction in procollagen production and increase in MMP-1 production. More precisely, EEB (50 ㎍/ml) markedly suppressed mRNA and protein levels of MMP-1 and -3. The anti-aging effects are attributable to the antioxidant activity of EEB. These findings indicate that EEB has a protective effect against UVB-induced aging in human fibroblast cells by regulating the levels of type-1 procollagen, MMP-1, and MMP-3.

• Childhood Kidney Diseases
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• v.7 no.1
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• pp.9-15
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• 2003

Purpose : Several studies have suggested that hyperlipidemia might be a causative factor contributing to the progression of initial glomerular injury through the development of glomerulosclerosis. We examined the potential beneficial effect of atorvastatin - which blocks the rate limiting step of cholesterol synthesis by inhibiting HMG-CoA reductase - in PAN-induced nephrosis. Materials and Methods : Glomerulosclerosis was induced in Sprague-Dawley male rats by repeated administration of PAN. Sprague-Dawley male rats were divided into 3 groups : group I(control), group II(PAN 20 mg/kg, subcutaneous injection), group III(PAN 20 mg/kg subcutaneous injection and atorvastatin 50 mg/kg/day per oral). On the 11th week, upon sacrifice of the experimental animals, blood sampling, 24-hr urine collection and nephrectomy were performed. Results : Group III had significantly lower BUN and higher serum albumin($30.9{\pm}17.2\;vs.\;17.3{\pm}2.5\;mg/dL;\;2.3{\pm}0.1\;vs.\;2.5{\pm}0.2\;g/dL$, P<0.05) compared with group II. In the lipid profiles, group III was associated with a reduction in total cholesterol and LDL($291{\pm}173\;vs.\;167{\pm}72\;mg/dL:\;57{\pm}53\;vs.\;27{\pm}12\;mg/dL$, P>0.05) compared with group II. Atorvastatin administration lowered the glomerular sclerosing index significantly(26.2% vs. 13.3%, P<0.05). Conclusion : Puromycin-induced glomerulosclerosis could be ameliorated by the reduction of hyperlipidemia with atorvastatin. This suggests that hyperlipidemia contributes to the pathogenesis of glomerulosclerosis.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.323-328
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• 2003

Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marshes and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display. Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86％ identity with Arabidopsis and 78％ with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCl concentrations.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.501-507
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• 1997

Chloroplasts isolated from Stevia rebaudiana Bertoni leaves contained an enzyme activity which catalyzed hydroxylation of ent-kaurenoic acid (ent-kaur-16-en-19-oic acid; ent-KA) to steviol (ent-13-hydroxy kaur-16-en-19-oic acid), the diterpenoid carboxylic alcohol which is the aglycone of sweet stevioside-related glycosides. $[^(14)C]-methylated$ ent-KA was used to localize ent-KA hydroxylase. $[^(14)C]-methyl-KA$ was most actively was transformed into methyl-steviol in chloroplast. The enzymatic activity was found in stroma fraction but not in thylakoid membrane in Stevia rebaudiana Bertoni. However, ent-KA 13-hydroxylase activity was not detected in stroma fraction of either Spinacia oleracea and Solidago altissima. The reaction products using $[^(14)C]-methyl-KA$ were purified and identified on TLC autoradiogram. The hydroxylation of ent-KA from stromal protein to form steviol required NADPH and oxygen. FAD and riboflavin stimulated the enzyme activity 1.5-and 1.7-fold, respectively. It also turned out that the activity of this enzyme using methyl-KA as a substrate was 16.7% that of ent-KA. The purified ent-KA 13-hydroxylase did not act on t-cinnamic acid, 4-hydroxyphenyl acetic acid, choline and resorcinol, known as monooxygenase and hydroxylase substrates.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.46 no.4
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• pp.303-308
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• 2001

To investigate the partitioning of newly absorbed N derived from NO$_3$- and NH$_4$$^{+}$, 6 mM $K^{15}$ NO$_3$ or 3 mM ($^{15}$ NH$_4$)$_2$ was fed continuously in Italian ryegrass (Lolium multiflrum L.) for 7 days. Nitrogen metabolites (nitrate, amino acid, soluble- and insoluble protein) were analyzed at the end of $^{15}$ N feeding. Dry weight in shoot, stubble and root was not significantly different between NO$_3$$^{[-10]}$ and NH$_4$$^{+}$ feeding. Total nitrogen content in all three organs was significantly higher in NH$_4$$^{+}$ than NO$_3$$^{[-10]}$ feeding. Sum on N content in reduced N fractions (amino acids + proteins) in shoot, stubble and roots in NH$_4$$^{+}$ feeding increased by 13.3, 12.5 and 35.4 %, respectively, compared to NO$_3$$^{[-10]}$ feeding. The Relative Specific Activity (RSA, percentage of newly absorbed $^{15}$ N relative to total N in a sample) values of amino acids and insoluble proteins were significantly higher in NH$_4$$^{+}$ feeding. Total amount of newly absorbed $^{15}$ N in NO$_3$$^{[-10]}$ and NO$_3$$^{[-10]}$ feeding was 52.3 and 69.5 mg/plant on dry matter basis, respectively. In both NH$_4$$^{+}$ and NO$_3$$^{[-10]}$ grown plants, most of the N was allocated to the shoot, 67.5% in NH$_4$$^{+}$ feeding and 58.8% NO$_3$$^{[-10]}$ feeding, respectively. The $^{15}$ N amount incorporated in the reduced N compounds (amino acids and proteins) in NH$_4$$^{+}$ grown plants significantly increased by 74.8% compared to NO$_3$$^{[-10]}$ grown plants. The increase of the $^{15}$ N amount assimilated to amino acids in NH$_4$$^{+}$ grown plants was remarkably higher in roots as more than 7.25 times compared to NO$_3$$^{[-10]}$ feeding. These results indicated that Italian ryegrass was much efficiently utilized NH$_4$$^{+}$-N for the synthesis of reduced N compounds.reduced N compounds.

• Restorative Dentistry and Endodontics
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• v.18 no.2
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• pp.261-276
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• 1993

The purpose of this study was to evaluate for the cytotoxicity of glass-ionomer cement liners(GC liningcement, Ketac-bond, Vitrebond and Fuji lining LC) on the fibroblasts cultured from human pulp. The fibroblasts were cultured in DMEM-10% FBS medium. The measurement of pH, succinate dehydrogenase (SDH) activity test and $^{51}Chromium$ release test were performed. Viable cell count and $^{14}C$-leucine incorporation rate were evaluated following culture time of 2, 4 and 6 days. The results of this study were as follows : 1. The pH in all cements was to be neutralized as time elapsed, and Fuji lining LC was the lowest pH value among them. 2. SDH activity was more inhibited in GC lining cement and Vitrebond than Ketac-bond and Fuji lining LC with the setting process, and GC lining cement and Ketac-bond were reduced after 5 minute's setting and then elevated as time elapsed. 3. In SDH activity test following exposure time, the activity in Vitrebond, GC lining cement and Fuji lining LC was inhibited with increased exposure time, but it was fairly constant in Ketac-bond. 4. Overall the liquid component was more inhibited than the powder component of glass-ionomer cement in SDH activity test. 5. In $^{51}Cr$-release test, Fuji lining LC was the most released of all the cements tested and followed by : Vitrebond, Ketac-bond, GC lining cement. 6. In viable cell count, the number of cells increased as the culture day proceeded in Ketac-bond, but they decreased in GC lining cement. Fuji lining LC was only observed after 2 days culture and there was not observed the whole culture days in Vitrebond. 7. In $^{14}C$-leucine incorporation rate test, protein synthesis was decreased with the number of culture days in GC lining cement, Vitrebond and Fuji lining LC, but it was followed that of control in Ketacbond.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.574-582
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• 1992

Exo-xylanase encoded by the xylA gene of Bacillus stearothermoPhillus was produced from Escherichia coli ]M109 carrying a recombinant plasmid pMGL Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at $37^{\circ}C$ for 8 hours on the medium containing 0.5% fructose, 1.0% tryptone, 1.0% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-IOO gel filtration, and Sephadex G-150 gel filtration. The' purified enzyme was most active at pH 6.0 and $45^{\circ}C$. $Ca^{2+}$ and $Co^{2+}$ activated the exo-xylanase activity by about 20% while $Ag^{2+}$, $Fe^{2+}$, $Mg^{2+}$ and $Zn^{2+}$ inhibited the enzyme activity by up to 60%. The $K_m$, value on p-nitrophenyl-$\beta$-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 daL SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 da!. polypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.