• Title/Summary/Keyword: fig enzyme

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Optimized Condition of Genomic DNA Extraction and PCR Methods for GMO Detection in Potato (유전자재조합 감자의 검정을 위한 DNA분리 및 PCR검출의 최적조건 탐색)

  • Shin, Weon-Sun;Kim, Myung-Hee
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.591-597
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    • 2003
  • To compare the quality of genomic DNA extracted from potato for PCR detection, four different methods, such as silica-based membrane method, silica-coated bead method, STE solution treatment, and CTAB-phenol/chloroform method, were evaluated. Also, to remove an excessive carbohydrate from the potato, ${\alpha}$- and ${\beta}$-amylase were used individually and in combination. When used both silica-based membrane method and silica-coated bead method combined with enzymes, the genomic DNAs were extracted from the raw potato with high purity for PCR. However, the silica-coated head method combined with enzyme treatment was the most efficient for extraction of the genomic DNA from the frozen fried potatoes. When applied with STE solution, the highly purified DNA was extracted from the raw potatoes without enzyme treatment in adequate yield for PCR. In cases of processed potatoes, such as frozen-fried potato and fabricated potato chips, CTAB-phenol/chloroform method is mostly feasible for DNA extraction and PCR efficacy at high sensitivity. As the results of PCR amplification, 216bp of PCR product was detected on 2% agarose gel electrophoresis, but any amplicons derived from New leaf and New leaf Y gene was not detected in any sample.

The Role of Tumor Necrosis Factor-$\alpha$ and Interleukin-$1{\beta}$ as Predictable Markers for Development of Adult Respiratory Distress Syndrome in Septic Syndrome (패혈증 증후군환자에서 성인성 호흡곤란 증후군 발생의 예측 지표서의 혈중 Tumor Necrosis Factor-$\alpha$와 Interleukin-$1{\beta}$에 관한 연구)

  • Koh, Youn-Suck;Jang, Yun-Hae;Kim, Woo-Sung;Lee, Jae-Dam;Oh, Soon-Hwan;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.452-461
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    • 1994
  • Background: Tumor necrosis factor(TNF)-$\alpha$ and Interleukin(lL)-$1{\beta}$ are thought to play a major role in the pathogenesis of the septic syndrome, which is frequently associated with adult respiratory distress syndrome(ARDS). In spite of many reports for the role of TNF-$\alpha$ in the pathogenesis of ARDS, including human studies, it has been reported that TNF-$\alpha$ is not sensitive and specific marker for impending ARDS. But there is a possibility that the results were affected by the diversity of pathogenetic mechanisms leading to the ARDS because of various underlying disorders of the study group in the previous reports. The purpose of the present study was to evaluate the roles of TNF-$\alpha$ and IL-$1{\beta}$ as a predictable marker for development of ARDS in the patients with septic syndrome, in which the pathogenesis is believed to be mainly cytokine-mediated. Methods: Thirty-six patients of the septic syndrome hospitalized in the intensive care units of the Asan Medical Center were studied. Sixteens suffered from ARDS, whereas the remaining 20 were at the risk of developing ARDS(acute hypoxemic respiratory failure, AHRF). In all patients venous blood samples were collected in heparin-coated tubes at the time of enrollment, at 24 and 72 h thereafter. TNF-$\alpha$ and IL-$1{\beta}$ was measured by an enzyme-linked immunosorbent assay (ELISA). All data are expressed as median with interquartile range. Results: 1) Plama TNF-$\alpha$ levels: Plasma TNF-$\beta$ levels were less than 10pg/mL, which is lowest detection value of the kit used in this study within the range of the $mean{\pm}2SD$, in all of the normal controls, 8 of 16 subjects of ARDS and in 8 in 20 subjects of AHRF. Plasma TNF-$\alpha$ levels from patients with ARDS were 10.26pg/mL(median; <10-16.99pg/mL, interquartile range) and not different from those of patients at AHRF(10.82, <10-20.38pg/mL). There was also no significant difference between pre-ARDS(<10, <10-15.32pg/mL) and ARDS(<10, <10-10.22pg/mL). TNF-$\alpha$ levels were significantly greater in the patients with shock than the patients without shock(12.53pg/mL vs. <10pg/mL) (p<0.01). There was no statistical significance between survivors(<10, <10-12.92pg/mL) and nonsurvivors(11.80, <10-20.8pg/mL) (P=0.28) in the plasma TNF-$\alpha$ levels. 2) Plasma IL-$1{\beta}$ levels: Plasma IL-$1{\beta}$ levels were less than 0.3ng/mL, which is the lowest detection value of the kit used in this study, in one of each patients group. There was no significant difference in IL-$1{\beta}$ levels of the ARDS(2.22, 1.37-8.01ng/mL) and of the AHRF(2.13, 0.83-5.29ng/mL). There was also no significant difference between pre-ARDS(2.53, <0.3-8.34ngfmL) and ARDS(5.35, 0.66-11.51ng/mL), and between patients with septic shock and patients without shock (2.51, 1.28-8.34 vs 1.46, 0.15-2.13ng/mL). Plasma IL-$1{\beta}$ levels were significantly different between survivors(1.37, 0.4-2.36ng/mL) and nonsurvivors(2.84, 1.46-8.34ng/mL). Conclusion: Plasma TNF-$\alpha$ and IL-$1{\beta}$ level are not a predictable marker for development of ARDS. But TNF-$\alpha$ is a marker for shock in septic syndrome. These result could not exclude a possibility of pathophysiologic roles of TNF-$\alpha$ and IL-$1{\beta}$ in acute lung injury because these cytokine could be locally produced and exert its effects within the lungs.

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Changes of the Antioxidant Enzymes and Histopathological Changes of Liver and Kidney According to the Administered Dosage of Bambusae Caulis in Liquamen in Mice (죽력 (Bambusae Caulis in Liquamen)의 투여량에 따른 생쥐의 항산화효소 활성과 간과 신장의 조직병리학적 변화)

  • Kim, Jeong-Sang
    • Applied Microscopy
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    • v.32 no.4
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    • pp.399-410
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    • 2002
  • This study was performed to investigate the effects of Bambusae Caulis in Liquamen (bamboo extract) on the changes of antioxidant enzymes and histopathological changes of liver and kidney in mice when administered in different dosages. The experimental groups were divided into four. For control group, 0.9% NaCl (0.2 ml/25 g B.W.) and for experimental groups, 5% (H1 group), 10% (H2 group), and 20%(H3 group) bamboo extract diluted with 0.9% NaCl, were administered (0.2 ml/25 g B.W.) respectively for 28 days at an interval of 48 hours. MnSOD activities were increased in H1 group (46%, P<0.05), H2 group (40%, P<0.05), and H3 group (34%, P<0.05) as compared to the control group. CuZnSOD activities were increased in H1 group (11%, P<0.05), but were decreased in H3 group (13%, P<0.05). The activities of catalase were decreased in H1 group (39%, P<0.05), H2 group (34%, P<0.05), and H3 group (31%, P<0.05) as compared to the control group. Histopathological observation revealed ballooned hepatocytes in the pericentral and periportal veins of H1 group. More ballooned and injured hepatocytes than in H1 group were observed in the H2 and H3 groups. Detachment of endothelial cells of the central vein was observed in the H2 and H3 groups. These results indicated that bamboo extract developed dose-dependent changes in antioxidant enzyme activities and developed histopathological changes of liver and kidney.

Studies on the Digestion of Beef by Ficin Treatment (Ficin 처리 우육의 소화에 관한 연구)

  • Kim, Jung-Sook;Kim, Jun-Pyong
    • Applied Biological Chemistry
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    • v.30 no.3
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    • pp.210-218
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    • 1987
  • In the previous report, we isolated and purified one of tendering enzyme 'ficin' from fig latex. In this study, various crude ficin concentrations and reaction time were employed to investigate the contents of free amino acids and other free nitrogen compounds after the treated with beef round muscle. 1. Free amino acids contents increased with the increase of temperature and time during the aging of beef at $1^{\circ}C$ and $8^{\circ}C$, and the increasing rate was remarkably high when fresh beef was treated with ficin. In the case of ficin treatment after various cooking, steaming showed the highest increase in free amino acid contents among three cooking methods such as boiling, steaming and pan broiling. The increased amounts of free amino acids in three groups-aging beef at $1^{\circ}C$ for 3 days, fresh beef treated with ficin(0.1%, 2hrs) and beef treated with ficin(0.1%, 2hrs) after cooking were 13%, 293% and 137% respectively. In contrast to aging group, the amount of free amino acids in other two groups treated with ficin was superiorly increased. 2. The amounts of total free nitrogen, free non-protein nitrogen and $NH_3-nitrogen$ increased with the increase of temperature and time during the aging of beef at $1^{\circ}C$ and $8^{\circ}C$, and the increasing rate was remarkably high in fresh beef treated with ficin. In the case of ficin treatment after cooking, steaming gave larger amount of total and non-protein nitrogen than other two cooking, e.g. boiling and panbroiling. The increasing rate of nonprotein nitrogen to the total nitrogen of fresh beef treated with ficin(0.1%, 2hrs) was 75 times greater than that of aging fresh beef at $1^{\circ}C$ for 3 days.

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Mechanism Underlying the Anti-Inflammatory Action of Piceatannol Induced by Lipopolysaccharide (당지질로 유도한 염증반응에서 Piceatannol의 항염증 기전 연구)

  • Cho, Han-Jin;Shim, Jae-Hoon;So, Hong-Seob;YoonPark, Jung-Han
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.9
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    • pp.1226-1234
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    • 2012
  • 3,4,3',5'-Tetrahydroxy-trans-stilbene (piceatannol) is a derivative of resveratrol with a variety of biological activities, including anti-inflammatory, anti-proliferative, and anti-cancer activities. We assessed the mechanisms by which piceatannol inhibits inflammatory responses using lipopolysaccharide (LPS)-treated Raw264.7 murine macrophages. Piceatannol (0~10 ${\mu}mol/L$) decreased LPS-induced release of nitric oxide, tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, IL-$1{\beta}$, and inhibited LPS-induced protein expression of inducible nitric oxide synthase (iNOS). Activation of nuclear factor-kappaB (NF-${\kappa}B$), activator protein (AP)-1, and signal transducer and activator of transcription 3 (STAT3) are crucial steps during an inflammatory response. Piceatannol prevented LPS-induced degradation of inhibitor of ${\kappa}B$ ($I{\kappa}B$), translocation of p65 to the nucleus, and phosphorylation of stress-activated protein kinase/c-Jun NH2-terminal kinase (SAPK/JNK). Additionally, piceatannol inhibited LPS-induced phosphorylation of STAT3 and IL-6-induced translocation of STAT3 to the nucleus. Furthermore, piceatannol increased the protein and mRNA levels of hemeoxygenase (HO)-1, the rate-limiting enzyme of heme catabolism that plays a critical role in mediating antioxidant and anti-inflammatory effects. Piceatannol further induced antioxidant response elements (ARE)-driven luciferase activity in Raw264.7 cells transfected with an ARE-luciferase reporter construct containing the enhancer 2 and minimal promoter region of HO-1. These results suggest that piceatannol exerts anti-inflammatory effects via the down-regulation of iNOS expression and up-regulation of HO-1 expression.

Study of the mechanisms underlying increased glucose absorption in Smilax china L. leaf extract-treated HepG2 cells (청미래덩굴 잎 물추출물이 처리된 HepG2 세포에서의 포도당흡수기전 연구)

  • Kang, Yun Hwan;Kim, Dae Jung;Kim, Kyoung Kon;Lee, Sung Mee;Choe, Myeon
    • Journal of Nutrition and Health
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    • v.47 no.3
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    • pp.167-175
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    • 2014
  • Purpose: Previous studies have shown that treatment with Smilax china L. leaf extract (SCLE) produces antidiabetic effects due to ${\alpha}$-glucosidase inhibition. In this study, we examined the mechanism underlying these antidiabetic effects by examining glucose uptake in HepG2 cells cultured with SCLE. Methods: Glucose uptake and glucokinase activity were examined using an assay kit. Expression of glucose transporter (GLUT)-2, GLUT-4, and HNF-$1{\alpha}$ was measured by RT-PCR or western blot. Results: Treatment with SCLE resulted in enhanced glucose uptake in HepG2 cells, and this effect was especially pronounced when cells were cultured in an insulin-free medium. SCLE induced an increase in expression of GLUT-2 but not GLUT-4. The increase in the levels of HNF-$1{\alpha}$, a GLUT-2 transcription factor, in total protein extract and nuclear fraction suggest that the effects of SCLE may occur at the level of GLUT-2 transcription. In addition, by measuring the change in glucokinase activity following SCLE treatment, we confirmed that SCLE stimulates glucose utilization by direct activation of this enzyme. Conclusion: These results demonstrate that the potential antidiabetic activity of SCLE is due at least in part to stimulation of glucose uptake and an increase in glucokinase activity, and that SCLE-stimulated glucose uptake is mediated through enhancement of GLUT-2 expression by inducing expression of its transcription factor, HNF-$1{\alpha}$.

Whitening activity of Ficus carica L. fruits extract through inhibition of tyrosinase and MITF expression (무화과(Ficus carica L.) 열매 추출물의 tyrosinase 및 MITF 발현 억제를 통한 미백 활성)

  • Min Ji Kim;Si Eun Park;Geun soo Lee;Jin Hwa Kim;Sunwoo Kwon;Hyung Seo Hwang
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.204-212
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    • 2023
  • Whitening is inhibitory activity of the melanin synthesis of melanocytes. Recently, whitening materials have been developed on natural materials because of its side effects on skin. Figs (Ficus Carica L.) is a fruit belonging to the Moraceae family and whitening activity was reported in focusing on the fig's stem and leaf components, but whitening activity of the figs fruit was not known. Thus, in this study, we tried to observe its anti-melanogenesis as well as antioxidant and anti-inflammation. The radical scavenging activity of figs fruits extract (FFE) was observed as the level of 34.52±1.98%/60.71±1.26% compared to the control in the its maximum concentration in the DPPH/ABTS assay. Cytotoxicity of FFE was observed at 10% concentration by CCK8 assay, so the maximum concentration was set at 5% and applied to all experiments. FFE concentration dependently decreased NO production associated with inducible nitric oxide synthase, cyclooxygenase-2, interleukin-6 and tumor necrosis factor-α gene expression, these strongly suggesting anti-inflammatory activity. In melanin contents assay, FFE significantly down-regulated melanin production in α-MSH-stimulated B16F10 cell as well as tyrosinase inhibition in vitro. In addition, FFE decreased the Microphthalmia-associated transcription factor (MITF) mRNA expression about 94.34% compared to the α-MSH treatment group in RT-PCR. Finally, FFE significantly reduced the MITF, cAMP response element-binding protein and tyrosinase protein expression in the α-MSH stimulated B16F10 cell. Through these results, we found that FFE can not only directly inhibit tyrosinase enzyme activity but also suppress melanogenesis through regulation of MITF gene expression in α-MSH signal transduction.

Studies on the effect of phthalimido methyl-O,O-dimethyl-phosphorodithioate (Imidan) and its possible metabolites on the growth of rice plant (Phthalimido methyl-O,O-dimethyl phosphorodithioate (Imidan)과 그의 대사물질(代謝物質)이 수도(水稻) 생육(生育)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Lee, Sung-Hwan;Lee, Dong-Suk;Lee, Jae-Koo
    • Applied Biological Chemistry
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    • v.7
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    • pp.105-117
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    • 1966
  • This experiment was conducted to investigate the effet of phthalimido-methyl-O,O-dimethyl-phosphorodithioate (Imidan) known as an acaricide and its possible metabolic products on the growth of plant, when sprayed on the leaves of rice plant. The results are summarized as follows. 1) Possible metabolic products of Imidan, the following compounds were synthesized or recrystallized for the present experiment a) N-Hydroxymethyl phthalimidem b) Phthalimide c) Phthalamidic acid d) Phthalic acid e) Anthranilic acid f) p-Amino benzoic acid g) p-Hydroxy benzoic acid h) Benzoic acid 2) Among the above materials, a), c), d), e), and Imidan were dissolved in a buffer solution respectively to be 10 and 20 p.p.m. and tested with the wheat coleoptile straight growth method. According to the results, Imidan inhibited the growth of coleoptile in both 10 and 20 p.p.m., whereas the others showed much better growth than the control, especially phthalamidic acid in 10 p.p.m. It appears that Imidan itself inhibits the coleoptile growth, whereas the metabolites derived from Imidan through various metabolisms, including hydrolysis in plant tissues show growth-regulating activity. (refer: Table 1, Fig. 1) 3) 20, 100 and 200 p.p.m. solutions of Imidall emulsion in xylene f·ere prepared. The lengths of shoot and root of rice seeds germinated on the re-respective media were measured after 12 days. The data showed that root was much more elongated in Imidan 20 p.p.m., whereas shoot in Imidan 100 p.p.m., respectively, than in the xylene control. An interesting finding was that xylene used as solvent had a tendency to inhibit seriously the root growth of rice seed. (refer: Table 2,5). 4) The emulsions of concentrations in 10, 25, 50 and 100 p.p.m's of control, Imidan, N-hydroxy methyl phthalimide, anthranilic acid, and phthalmide, respectively, were sprayed twice on the rice plant on pot. After a certain period of time lengths of rice culms were measured, showing that plots treated with Imidan and N-hydroxy methyl phthalimide exhibited much more growth than those of control and the others. 5) Loaves and stems of rice plant were sampled and extracted with dried acetone at the intervals of 3-, 5-, 7-, and 14 days after treated with Imidan 250 p.p.m. emulsion. This sample extracted with acetone was purified by means of prechromatographic purification method with acetonitrile and paperchromatographed to detect the following metabolic products. Imidan (Rf: 0.97-0,98), N-hydroxy-methyl phthalimide (Rf: 0.87) phthalimide (Rf: 0.86-0.87), phthalamidic acid (Rf: 0.13-0.14), phthalic acid (Rf: 0.02-0.03), benzoic acid (Rf: 0.42-0.43), p-amino benzoic acid or p-hydroxy benzoic acid (Rf: 0.08-0.09), and unidentified compounds (Rf: 0.73, 0.59, 0.33, 0.23. 0.07). In addition, in the early stages, such as 3- and 5 days nonhydrolyzed Imidan and its first hydrolytic product, N-hydroxymethyl phthalimide were detected in relatively large amounts, whereas in the last stages of 7- and 14 days due to further decomposition, the afore-mentioned two materials were reduced in the amount and p-hthalic, phthalamidic, benzoic, and p-Hydroxy benzoic, or p-Amino benzoic acids were detected in a considerably large amount. It is, therefore, believed that most of Imidan applied to the leaves of rice plant may be decomposed within almost 14 days. In the light of above observations it is considered that Imidan itself is not involved in plant growth regulating activity, whereas various phthaloyl derivatives produced in the course of metabolism (namelr, enzymic action) in plant tissues may have such effect.

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Studies on the Organo-mercury Residues in Rice Grain -I. Mercury residues in rice seeds treated with organo-mercury fungicide- (수도(水稻)에 처리(處理)된 유기수은제(有機水銀劑)의 잔류성(殘留性)에 관(關)한 연구(硏究) -제1보(第1報) 침지용유기수은제(浸漬用有機水銀劑)로 처리(處理)된 수도종자중(水稻種子中)의 수은잔류량(水銀殘留量)에 관(關)하여-)

  • Lee, Dong-Suk
    • Applied Biological Chemistry
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    • v.8
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    • pp.87-93
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    • 1967
  • 1) For the micro-analysis of mercury in plant materials, the method of Furutani was shown to be the simplest and most efficient way and the recovery of the assay was about 98%. 2) When the rice grain was soaked in 1/1000 diluted solution of organo-mercury fungicide for 8 hours at the end of March, the amounts of mercury residues in the brown rice and unhulled rice were 8.8 to $9.5\;{\mu}g/g$ seeds and 10.1 to $10.7\;{\mu}g/g$ seeds, respectively. 3) By washing the treated rice seeds with running water for three days, tile residual mercury concentration was reduced to 1/4 to 1/5; thus the mercury residues were 1.86 to $1.92\;{\mu}g/g$ for brown rice and 1.96 to $2.93\;{\mu}g/g$ for unhulled rice. 4) The residual mercury was present more in the unhulled rice than in the brown rice, either before or after washing of the treated seeds. 5) Among the different rice varieties, no difference was observed in mercury residues by seed treatment and washing.

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